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Background The cadherin-4 gene (CDH4), a member of the cadherin family genes, encodes R-cadherin (R-cad); however, the function of this gene in different types of cancer remains controversial. The function of CDH4 in OSCC (oral squamous cell carcinoma) is unknown. Materials and methods We use the Cancer Genome Atlas (TCGA) database to find the expression of CDH4 in OSCC is more than normal tissue. Our tissue samples also confirmed that CDH4 gene was highly expressed in OSCC. The related cell function assay detected that CDH4 promotes the ability of cell proliferation, migration, self-renewal and invasion. Cell staining experiment confirmed that the change of CDH4 expression would change the cell mortality. The western blot of GPX4 (glutathione-dependent peroxidase-4), GSH (reduced glutathione) test assay and MDA(Malondialdehyde) test assay show that the expression of CDH4 may resist the sensitivity of ferropotosis in OSCC. Results CDH4 was upregulated in OSCC samples and was correlation with poor survival of patients. High expression of CDH4 effectively promotes the proliferation, mobility of OSCC cells and reduce the sensitivity of OSCC cells to ferroptosis. CDH4 is positively correlated with EMT pathway genes, negatively correlated with fatty acid metabolism pathway genes and peroxisome pathway genes, and positively correlated with ferroptosis suppressor genes in OSCC. Conclusions These results indicate that CDH4 may play a positive role in tumor progression and resistance ferroptosis and may be a potential therapeutic target for OSCC.

The hilly region in eastern Guangdong, China lacks comprehensive scientific investigations for decades, especially in terms of herpetofauna. In recent years, several highly threatened amphibians have been gradually discovered from this region. In this work, three new species of the genus Boulenophrys are described, which are endemic from only one or two known localities in eastern Guangdong. These discoveries enrich the diversity of Boulenophrys in eastern Guangdong. With the large number of threatened urodeles and anuran species occurring in this densely populated area, the unique herpetological diversity in eastern Guangdong is facing the impacts of habitat degradation and fragmentation, and conservation actions are urgently required.

ObjectiveTo study specific information on trends in incidence, mortality, disability-adjusted life years (DALY) and the corresponding trends among five sociodemographic index regions, 21 regions, and 204 countries for decision-making, which would enable policymakers to distribute limited resources and devise policies more rationally.MethodsData on uterine fibroids (UNs) from 1990 to 2019, including incidence, mortality, and DALYs, were obtained from the 2019 Global Burden of Disease Study. An estimated annual percentage change (EAPC) was calculated to assess morbidity, mortality, and DALY trends.ResultsThe incident cases of UFs increased from 5,769,658 (95%UI, 7,634,3995–4,274,824) incidences in 1990 to 9,643,336 (95%UI, 7,178,053-12,714,741) incidences in 2017, and the age-standardized incidence rate was steady at 225.67/100,000 persons (95%UI, 167.33–298.87) in 1990 to 241.18/100,000 persons (95%UI, 179,45–318.02) in 2019. The incidence ratio in the high sociodemographic index (SDI) region showed a unimodal distribution, with peaks in 2005. Between 2009 and 2017, the age-standardized death rate of UFs declined globally, especially in low-SDI and low-middle SDI regions. In contrast with 860,619 DALYs (95%UI, 473,067-1,505,289) in 1990, the number of DALYs was 1,378,497 (95%UI, 710,915-2,475,244) in 2019, which had increased significantly, whereas the age-standardized DALY rate decreased expressively with an EAPC of −1.93 (95%CI, from −2.16 to −1.71).ConclusionThe global burden of UFs increased between 1990 and 2019, and the incidences and DALYs increased prominently worldwide, while the deaths from UFs had no evident growth. Lower SDI regions carried an incremental burden of UFs, while disease reduction was observed in higher SDI regions. It is high time we paid attention to the underprivileged regional quality of life and health protection.

BackgroundSpontaneous splenic rupture (SSR) is a rare, often life-threatening, acute abdominal injury that requires immediate diagnosis and early treatment. SSR is mainly treated surgically or conservatively. A few cases of interventional embolization for SSRs have been reported.Case PresentationA 30-year-old male patient complaining mainly of left upper abdominal pain underwent emergency abdominal computed tomography (CT) and showed enlargement of the spleen with a massive mixed-density shadow approximately 10.0 × 8.0 × 12.5 cm in size. The boundary was unclear and showed obvious progressive enhancement. Considering the intrasplenic tumor lesions with rupture and hemorrhage, the possibility of vascular tumors was high, with intraperitoneal blood and fluid accumulation. Digital subtraction angiography of the splenic arteriography and embolization of the ruptured splenic artery branches were performed. Postoperative hemoglobin progressively decreased, inflammatory indicators, such as white blood cell counts, procalcitonin (PCT), and C-reactive protein (CRP) were significantly increased, and 2 days after embolization, the patient developed severe hypoxemia, shock, pulmonary edema, and acute respiratory distress syndrome. CT re-examination 9 days after embolization showed reduced lesion absorption. After stabilization of the condition, splenectomy was performed, and postoperative platelet count increase, anticoagulant improvement, and discharge were observed. Postoperative pathological examination revealed extensive hemorrhage and necrosis, vascular tissue with abnormal hyperplasia in the surrounding area, vascular tissue in the bleeding area and outer wall (elastic fiber staining +), and local myofibroblast hyperplasia. Immunohistochemistry showed actin (SM +) and Ki67 (10% +).ConclusionSSR caused by splenic hemangioma is rare, and the choice between surgical treatment or splenic artery embolization remains dependent on the patient's hemodynamic stability and imaging findings.

Our previous study demonstrated a close relationship between the NOTCH signaling pathway and salivary adenoid cystic carcinoma (SACC). Its receptor gene, NOTCH1, and its downstream gene, HES1, contribute to the proliferation, invasion and metastasis of SACC. Accumulating evidence supports HEY1 as another effector of the signaling pathway. The purpose of this study was to explore the effects of the NOTCH1-HEY1 pathway on the proliferation, invasion and metastasis of SACC cells. Our results verified that HEY1 is a specific molecular target of the NOTCH signaling pathway in SACC cells and that its expression in carcinoma is much higher than that in paracarcinoma tissues. The expression of NOTCH1 and HEY1 are positively correlated in the salivary adenoid cystic carcinoma tissues. NOTCH1 is significantly related to the activation of HEY1 in SACC, and that HEY1 reciprocally regulates NOTCH1 expression in SACC. HEY1 promotes cell proliferation and spheroid formation and inhibits cell apoptosis in vitro. In addition, HEY1 enhances the tumorigenicity of SACC in vivo. Furthermore, HEY1 increases cell invasion and metastasis by driving the expression of epithelial-mesenchymal transition (EMT)-related genes and MMPs. The results of this study indicate that the NOTCH1-HEY1 pathway is specifically upregulated in SACC and promotes cell proliferation, self-renewal, invasion, metastasis and the expression of EMT-related genes and MMPs. Our findings suggest that a NOTCH1-HEY1 pathway inhibitor might therefore have potential therapeutic applications in treating SACC patients by inhibiting cancer cell growth and metastasis.

Boulenophrys fengshunensis sp. nov. Wang, Zeng, Lyu & Wang Fengshun Horned Toad (in English) / Fēng Shùn Jiǎo Chán (丰NJDzdz in Chinese) Chresonymy: Megophrys sp 14 — Liu et al. 2018 Figures 7–8 Holotype. SYS a004744, adult male, collected by Jian Wang and Zhi-Tong Lyu on 13 May 2016 from Mt. Tongguzhang (24°10'31.12"N, 116°21'2.63"E; ca. 1500 m a.s.l.), Fengshun, Meizhou, Guangdong, China. Paratypes (N=7). Adult males, SYS a004724/ CIB118528, SYS a004725–4728, collected by Jian Wang and Zhi-Tong Lyu on 11 May 2016 from the same locality of the holotype. Adult females, SYS a005220–5221, collected by Jian Wang on 9 August 2016 from the same locality of the holotype. Etymology. The specific epithet “ fengshunensis ” refers to the type locality of the new species, the Fengshun County. Diagnosis. (1) Small body size, SVL 34.3–39.4 mm (36.9 ± 1.7, N = 6) in adult males and SVL 42.5–44.9 mm (N = 2) in adult females; (2) snout pointed in dorsal view; (3) tympanum moderate in size TD/ED 0.56–0.67, tubercles bearing spines on tips present on skin of temporal region including the tympanum; (4) vomerine ridge and vomerine teeth present; (5) margin of tongue rounded, not notched distally; (6) hindlimbs short, heels not meeting and tibio-tarsal articulation reaching forward to the region between tympanum and posterior corner of eye, TIB/SVL 0.37–0.43; (7) a subarticular tubercle present at the base of each fingers; (8) toes without lateral fringes and with rudiment of webbing; (9) distinct sparse enlarged tubercles on the surface of limbs, flanks, posterior part of belly and around the cloaca; (10) tips of the tubercles on ventral surface of thighs and around the cloaca bearing tiny spines; (11) single subgular vocal sac in males; (12) nuptial pads with well-developed villiform black nuptial spines on the dorsal surface of the first and second fingers in breeding males. Comparisons. Comparative data of Boulenophrys fengshunensis sp. nov. and the other recognized members of Boulenophrys are listed in Table 3. Boulenophrys fengshunensis sp. nov. can be distinguished from Boulenophrys puningensis sp. nov. and Boulenophrys hungtai sp. nov. having a tympanum with tubercles bearing spines on their tips (vs. tympanic region smooth without granules, tubercles or spines in Boulenophrys puningensis sp. nov. and Boulenophrys hungtai sp. nov.); the presence of both a vomerine ridge and vomerine teeth (vs. absence of vomerine ridge but with vomerine teeth present in Boulenophrys hungtai sp. nov.); tubercles on skin of the posterior part of belly without spines (vs. tubercles on skin of the posterior part of belly bearing spines on their tips in both Boulenophrys puningensis sp. nov. and Boulenophrys hungtai sp. nov.). With a smaller body size, SVL 34.3–39.4 mm in adult males and SVL 42.5–44.9 mm in adult females, Boulenophrys fengshunensis sp. nov. differs from the seven congeners whose SVL ≥ 50 mm in males or SVL ≥ 60 mm in females, including B. caudoprocta (81.3 mm in a single male), B. jingdongensis (53.0– 56.5 mm in males and 63.5 mm in a single female), B. liboensis (60.5–67.7 mm in males and 60.8–70.6 in females), B. mirabilis (55.8–61.4 mm in males and 68.5–74.8 mm in females), B. omeimontis (56.0– 59.5 mm in males and 68.0– 72.5 mm in females), B. sangzhiensis (54.7 mm in a single male), and B. shuichengensis (102.0– 118.3 mm in males and 99.8–115.6 mm in females). In having relatively shorter shanks with heels that do not meet when the flexed hind limbs are held at right angles to the body axis, Boulenophrys fengshunensis sp. nov. can be easily distinguished from the following 35 congeners, i.e. B. angka, B. anlongensis, B. baishanzuensis, B. baolongensis, B. binchuanensis, B. binlingensis, B. boettgeri, B. congjiangensis, B. cheni, B. chishuiensis, B. daiyunensis, B. jiangi, B. jinggangensis, B. jiulianensis, B. leishanensis, B. lini, B. minor, B. mufumontana, B. nanlingensis, B. palpebralespinosa, B. qianbeiensis, B. sanmingensis, B. shimentaina, B. shunhuangensis, B. spinata, B. tongboensis, B. tuberogranulatus, B. wuliangshanensis, B. wushanensis, B. xiangnanensis, B. xianjuensis, B. yaoshanensis, B.yangmingensis, B. yingdeensis and B. yunkaiensis, all of which have relatively longer shanks with the heels meeting or overlapping. By the presence of vomerine teeth, Boulenophrys fengshunensis sp. nov. differs from B. acuta, B. brachykolos, B. caobangensis, B. daoji, B. huangshanensis, B. kuatunensis, B. lishuiensis, B. lushuiensis, B. obesa, B. ombrophila, and B. wugongensis, all of which lack vomerine teeth. By having a rounded margin of the tongue that is not notched distally, Boulenophrys fengshunensis sp. nov. differs from B. hoanglienensis, B. insularis, which have a notched tongue. By the absence of lateral fringes on toes, Boulenophrys fengshunensis sp. nov. differs from B. rubrimera, which has narrow lateral fringes on toes. By the presence of rudimentary webbing on toes, Boulenophrys fengshunensis sp. nov. differs from B. daweimontis, B. fansipanensis, B. frigida, and B. rubrimera, all of which lack webbing on toes. Boulenophrys fengshunensi sp. nov. further differs from the remaining B. dongguanensis and B. nankunensis by having raised tubercles on the surface of posterior abdomen (vs. absence of such tubercles in both B. dongguanensis B. nankunensis); the presence of tubercles bearing spines on their tips on the temporal region including the tympanum (vs. temporal region lacking tubercles or spines in both B. dongguanensis and B. nankunensis). Description of holotype. Adult male. Small body size, SVL 37.4 mm; head width slightly larger than head length, HWD/HDL 1.17; snout pointed in dorsal view, projecting, sloping backward to mouth in profile, protruding well beyond margin of lower jaw; top of head flat; eyes moderate in size, ED 0.37 of HDL, pupil vertical, near diamond-shaped; nostril oblique-ovoid; canthus rostralis well developed; loreal region slightly oblique; internasal distance slightly larger than interorbital distance; tympanum moderate in size with an obvious margin, TD/ED 0.65; large ovoid choanae at base of maxilla; vomerine ridge and vomerine teeth present, maxillary teeth present; margin of tongue rounded, not notched distally; presence of single subgular vocal sac, and pair of slit-like openings at posterior of jaw. Forearm (i.e., radioulna) length 0.21 of SVL and hand 0.25 of SVL; hand without webbing, fingers without lateral fringes, relative finger length I = II Dorsal skin rough, granular, with sparse tubercles; skin of dorsal body and limbs bearing tiny spines; sparse large conical tubercles on flanks; single horn-like prominent tubercle on edge of upper eyelid; obvious supratympanic fold curving posteroventrally from posterior corner of eye to level above insertion of arm; dense tubercles on skin of upper lip, upper eyelid, mandibular articulation, loreal, temporal region including tympanum and surface around cloaca; tip of tubercles on skin of temporal region excluding tympanum bearing tiny spine; single discontinuous “X” shaped ridge and few elongated tubercles arranged in longitudinal rows on two sides at mid-dorsum; sparse tubercles on dorsal shank and thigh; ventral skin of throat, chest and anterior part of belly smooth; ventral skin of thighs and posterior part of belly bearing dense tubercles; surface around cloaca with dense tubercles bearing tiny spines; small pectoral gland closer to axilla; single femoral gland positioned on posterior surface of thigh at midpoint between knee and cloaca.. Coloration of holotype in life. Dorsal surface of body dark brown, with incomplete dark brown triangular marking between eyes. Two wide oblique black bands present on forearm. Dorsal surface of fingers and hindlimbs with dark grey transverse bands. Presence of vertical dark brown band below eye. Tip of tubercles on edge of upper eyelid greyish white. Supratympanic fold orangey brown. Ventral surface dark brown, black longitudinal band on surface of throat, posterior part of belly with irregular greyish white patches. Tubercles on ventral surface of posterior part of belly and thighs greyish white; spines on tips of tubercles on temporal region, ventral surface of thighs and around cloaca dark grey. Digits, inner and outer metacarpal tubercles, and inner metatarsal tubercle dark grey. Pectoral glands beige and femoral glands greyish white. Iris orangish brown. Coloration of holotype in preservative. Dorsal surface of body dark brown. Triangular marking between eyes, vertical dark brown band below eye and transverse bands on dorsal forearms and hind limbs become indistinct. Supratympanic fold greyish white. All bands and patterns on ventral surface no longer apparent. Irregular greyish white patches on posterior part of belly become dark grey. Variation. Mensural data of the type series are listed in Table 6. Most of the paratypes are similar to the holotype in morphology and color pattern, except for the following: dorsal surface and ventral surface dark brown, posterior part of belly with irregular greyish white patches, ventral surface of thighs dark grey, triangular marking between eyes is incomplete in the holotype (vs. dorsal surface beige and ventral surface light orange, posterior part of belly and ventral surface of thighs greyish white, triangular marking between eyes is complete in the female paratype SYS a005221 (Fig. 8); tibio-tarsal articulation reaching forward to middle of tympanum when hind limb stretched along body in the holotype (vs. tibio-tarsal articulation reaching forward to posterior margin of tympanum in the paratype SYS a004725; reaching forward to the region between tympanum and eye in the paratype SYS a004726). Females are distinctly larger than the males. Distribution and natural history. Currently, Boulenophrys fengshunensis sp. nov. is only known from its type locality, Mt. Tongguzhang of Fengshun. It inhabits flowing montane streams and the nearby forest floor and leaf litter at elevations between 800–1500 m. Boulenophrys fengshunensis sp. nov. is found to be sympatric with Pachytriton granulosus Chang, 1933 and Cynops glaucus Yuan, Jiang, Ding, Zhang & Che, 2013. Advertisement calls of males were heard during April and June. Males were found calling under the leaf litters around the flowing seeps. Tadpoles could be found in this period. Female specimens collected during August had immature eggs; males were not heard calling during this period, but sub-adults were observed.

Boulenophrys hungtai sp. nov. Wang, Zeng, Lyu, Xiao & Wang Hung-Ta Chang���s Horned Toad (in English) / Jiē Y��ng Ji��o Ch��n (DZffiDzdz in Chinese) Figures 5���6 Holotype. SYS a007578, adult male, collected by Jian Wang, Hong-Hui Chen and Hui-Wen Xiao on 5 January 2019 from Mt. Liwangzhang (23��38'6.42"N, 115��48'51.78"E; ca. 990 m a.s.l.), Jiexi, Jieyang, Guangdong, China. Paratypes (N=12). SYS a007575/ CIB118527, SYS a007576���7577, 7579���7582, 7594���7597, adult males, collected on 5���6 January 2019 from the same stream as the holotype at elevations between 950���1000 m. SYS a008576, adult male, collected by Jian Wang, Hong-Hui Chen, and Shuo Qi on 28 February 2021 from Shuangkeng Forestry Station (23��43'56.25"N, 116��21'26.4"E; ca. 550 m a.s.l.), Jiedong, Jieyang, Guangdong, China. Etymology. The specific epithet ��� hungtai ��� is a patronym in honor of Professor Hung-Ta Chang (=Hong-Da Zhang, �����ffl, 1914���2016), an outstanding botanist who was born in Jiexi. Diagnosis. (1) Small body size, SVL 25.8���33.3 mm (28.2 �� 2.3, N = 12) in adult males; (2) snout pointed in dorsal view; (3) tympanum moderate, TD/ED 0.52���0.61; (4) tympanic region smooth without granules or tubercles; (4) vomerine ridge and vomerine teeth absent; (5) margin of tongue rounded, not notched distally; (6) hindlimbs short, heels not meeting and tibio-tarsal articulation reaching forward to the region between tympanum and posterior corner of eye; (7) a subarticular tubercle present at the base of each fingers; (8) toes without lateral fringes and webbing; (9) distinct enlarged tubercles on the surface of limbs, flanks, chest, belly and around the cloaca; (10) tips of the tubercles on posterior abdomen, ventral thighs and around the cloaca bearing tiny spines; (11) single subgular vocal sac in males; (12) nuptial pads with villiform black nuptial spines on the dorsal surface of the first and second fingers in breeding males. Comparisons. Comparative data of Boulenophrys hungtai sp. nov. and the other recognized members of Boulenophrys are listed in Table 3. Boulenophrys hungtai sp. nov. can be easily distinguished from Boulenophrys puningensis sp. nov. by having a moderately sized tympanum, TD/ED 0.52���0.61 (vs. tympanum large, TD/ED 0.68���0.71); lacking both a vomerine ridge and vomerine teeth (vs. vomerine ridge and vomerine teeth present); lacking webbing on toes (vs. having rudimentary webbing on toes). Boulenophrys hungtai sp. nov. is strongly supported as the sister taxon to B. insularis, which is stated by Wang et al. (2017a) to be an endemic species of an offshore island in Shantou, China. Though the mean p -distance in the 16S gene is only 2.2 %, B. hungtai differs by having a snout pointed in dorsal view (vs. snout obtusely rounded in dorsal view); by the absence of vomerine ridge and vomerine teeth (vs. presence of strong vomerine ridge bearing vomerine teeth); margin of tongue not notched distally (vs. tongue notched distally); absence of webbing on toes (vs. having rudiment of webbing on toes); distinct enlarged tubercles on the surface of limbs, flanks, chest, belly and around the cloaca (vs. ventral surface smooth). Boulenophrys hungtai sp. nov. forms a clade with B. ombrophila (mean p -distance 4.9 % in the 16S gene), B. obesa (mean p -distance 3.9 % in the 16S gene), and B. cheni (mean p -distance 2.5 % in the 16S gene) though there is weak support for these relationships. However, the new species differs from the latter two congeners by having no webbing on toes (vs. having rudiment of webbing on toes in B. obesa); no vomerine ridge (vs. presence of vomerine ridges in B. obesa); a pointed snout in dorsal view (vs. snout rounded in dorsal view in B. ombrophila and B. obesa); distinct enlarged tubercles on the surface of limbs, flanks, chest, belly and around the cloaca (vs. ventral surface smooth in B. ombrophila and B. obesa); margin of tongue not notched distally, the shorter shanks with heels not meeting, absence of lateral fringes on toes (vs. tongue notched, heels meeting, presence of wide lateral fringes on toes in B. cheni). With a smaller body size, SVL 25.8���33.3 mm in adult males, Boulenophrys hungtai sp. nov. differs from the seven congeners whose SVL ��� 50 mm in males, including B. caudoprocta (81.3 mm in a single male), B. jingdongensis (53.0��� 56.5 mm in males), B. liboensis (60.5���67.7 mm in males), B. mirabilis (55.8���61.4 mm in males), B. omeimontis (56.0��� 59.5 mm in males), B. sangzhiensis (54.7 mm in a single male), and B. shuichengensis (102.0��� 118.3 mm in males). In having relatively shorter shanks with heels that do not meet when the flexed hind limbs are held at right angles to the body axis, Boulenophrys hungtai sp. nov. can be easily distinguished from the following 31 congeners, i.e. B. angka, B. anlongensis, B. baishanzuensis, B. baolongensis, B. binchuanensis, B. binlingensis, B. boettgeri, B. congjiangensis, B. chishuiensis, B. jiangi, B. jinggangensis, B. jiulianensis, B. leishanensis, B. lini, B. minor, B. mufumontana, B. nanlingensis, B. palpebralespinosa, B. qianbeiensis, B. sanmingensis, B. shimentaina, B. shunhuangensis, B. spinata, B. tongboensis, B. wuliangshanensis, B. wushanensis, B. xiangnanensis, B. xianjuensis, B. yaoshanensis, B. yangmingensis, B. yingdeensis and B. yunkaiensis, all of which have relatively longer shanks with the heels meeting or overlapping. By the absence of vomerine teeth, Boulenophrys hungtai sp. nov. differs from B. daiyunensis, B. daweimontis, B. dongguanensis, B. fansipanensis, B. frigida, B. hoanglienensis, B. nankunensis, and B. rubrimera, all of which have vomerine teeth. By having a rounded margin of the tongue that is not notched distally, Boulenophrys hungtai sp. nov. differs from B. huangshanensis, B. kuatunensis, and B. lushuiensis, all of which have notched tongues. By the absence of lateral fringes on toes, Boulenophrys hungtai sp. nov. differs from B. acuta, and B. daoji, all of which have lateral fringes on toes. By the absence of webbing on toes, Boulenophrys hungtai sp. nov. differs from B. brachykolos, B. caobangensis, B. tuberogranulatus, and B. wugongensis, all of which have rudimentary webbing on toes. Boulenophrys hungtai sp. nov. further differs from the remaining B. lishuiensis by having raised tubercles bearing spines on their tips on surface of posterior abdomen (vs. surface of belly smooth in B. lishuiensis). Description of holotype. Adult male. Body size small, SVL 28.7 mm. Head width slightly larger than head length, HDW/HDL 1.11; snout pointed in dorsal view, projecting, sloping backward to mouth in profile, protruding well beyond margin of lower jaw; top of head flat; eyes moderate in size, ED 0.34 of HDL, pupil vertical, near diamond-shaped; nostril oblique-ovoid; canthus rostralis well developed; loreal region slightly oblique; internasal distance slightly larger than interorbital distance; tympanum moderate in size with an obvious margin, TD/ED 0.58; large ovoid choanae at base of maxilla; vomerine ridges and vomerine teeth absent, maxillary teeth present; margin of tongue rounded, not notched distally; presence of a single subgular vocal sac, and pair of slit-like openings at posterior of jaw. Forearm (i.e., radioulna) length 0.22 of SVL and hand 0.23 of SVL; hand lacking webbing, fingers lacking lateral fringes, relative finger length I Coloration of holotype in life. Dorsal surface of body maroon, with an incomplete dark brown triangular marking between eyes. Flanks yellowish brown. Two wide oblique black bands present on forearm. Dorsal surface of fingers and hindlimbs with dark grey transverse bands. Presence of vertical dark brown band below eye. Tip of tubercle on the edge of upper eyelid white. Supratympanic fold greyish white with orange mottling. Ventral surface of throat, chest, and sides of belly dark brown with white and orange mottling, black longitudinal band on surface of throat; central and posterior part of belly white, with dark brown patches and orange mottling. Tubercles on ventral surface of chest, belly, and thighs greyish white; spines on tips of tubercles on surface of posterior abdomen, ventral surface of thighs and around cloaca dark grey. Digits, inner and outer metacarpal tubercles, and inner metatarsal tubercle greyish white. Pectoral glands and femoral glands beige. Iris reddish brown. Coloration of holotype in preservative. Maroon fades to dark brown dorsally. Coloration of flanks fades to greyish brown. Triangular marking between eyes and transverse bands on dorsal forearms and hind limbs become indistinct. Orange mottling on the supratympanic fold absent. Color of ventral surface fades; patterns of become indistinct; orange mottling on ventral skin absent. Pectoral glands and femoral glands greyish white. Variation. Mensural data of the type series are listed in Table 5. Most of the paratypes are similar to the holotype in morphology and color pattern, except for the following: coloration of dorsum is light brown and iris is reddish brown in the holotype (vs. dorsum beige and iris greyish white with irregular dark brown and light orange patterns in the paratype SYS a008576 (Fig. 6A���B); central and posterior part of belly white, with dark brown patches and orange mottling in the holotype (vs. ventral skin dark brown, without regular patches in the paratype SYS a008576 (Fig. 6C���D); larger body size in the paratypes SYS a007582 (SVL 32.7 mm) and SYS 008576 (SVL 33.3 mm); tibio-tarsal articulation reaching forward to middle of tympanum when hind limb stretched along body (vs. tibio-tarsal articulation reaching forward to posterior corner of eye in the paratypes SYS a007576, 7582, 7596, 7597, 8576). Distribution and natural history. Currently, Boulenophrys hungtai sp. nov. is known from Mt. Liwangzhang of Jiexi (800���1000 m a.s.l.) and Shuangkeng Forestry Station (500���800 m a.s.l.) of Jiedong, which are ca. 55 km from each other. This toad inhabits flowing montane streams and the nearby forest floor and leaf litter, and is sympatric with Pachytriton brevipes (Sauvage, 1876). Cynops orphicus Risch, 1983 can also be observed in the surrounding area. Advertisement calls of males from Mt. Liwangzhang were heard from November to the following January. A single male (SYS a008576) from Shuangkeng Forestry Station was noticed discontinuously calling on March after the heavy rain. Males were found calling under the leaf litter or in rock crevices in flowing streams., Published as part of Wang, Jian, Zeng, Zhao-Chi, Lyu, Zhi-Tong, Qi, Shuo, Liu, Zu-Yao, Chen, Hong-Hui, Lu, Yu-Hong, Xiao, Hui-Wen, Lin, Can-Rong, Chen, Kai & Wang, Ying-Yong, 2022, Description of three new Boulenophrys species from eastern Guangdong, China, emphasizing the urgency of ecological conservation in this region (Anura, Megophryidae), pp. 91-119 in Zootaxa 5099 (1) on pages 106-110, DOI: 10.11646/zootaxa.5099.1.4, http://zenodo.org/record/6036964, {"references":["Wang, J., Liu, Z. Y., Lyu, Z. T., Zeng, Z. C. & Wang, Y. Y. (2017 a) A new species of the genus Xenophrys (Amphibia: Anura: Megophryidae) from an offshore island in Guangdong Province, southeastern China. Zootaxa, 4324 (3), 541 - 556. https: // doi. org / 10.11646 / zootaxa. 4324.3.8","Sauvage, H. E. (1876) L'Institut. Journal des Academies et Societes Scientifiques de la France et de l'Etrangers, Paris, 4, 274 - 275.","Risch, J. P. (1983) Cynops orphicus, a new salamander from Guangdong Province, South China (Amphibia, Caudata, Salamandridae). Alytes, Paris, 2, 45 - 52."]}

Boulenophrys fengshunensis sp. nov. Wang, Zeng, Lyu & Wang Fengshun Horned Toad (in English) / Fēng Sh��n Ji��o Ch��n (���NJDzdz in Chinese) Chresonymy: Megophrys sp 14 ��� Liu et al. 2018 Figures 7���8 Holotype. SYS a004744, adult male, collected by Jian Wang and Zhi-Tong Lyu on 13 May 2016 from Mt. Tongguzhang (24��10'31.12"N, 116��21'2.63"E; ca. 1500 m a.s.l.), Fengshun, Meizhou, Guangdong, China. Paratypes (N=7). Adult males, SYS a004724/ CIB118528, SYS a004725���4728, collected by Jian Wang and Zhi-Tong Lyu on 11 May 2016 from the same locality of the holotype. Adult females, SYS a005220���5221, collected by Jian Wang on 9 August 2016 from the same locality of the holotype. Etymology. The specific epithet ��� fengshunensis ��� refers to the type locality of the new species, the Fengshun County. Diagnosis. (1) Small body size, SVL 34.3���39.4 mm (36.9 �� 1.7, N = 6) in adult males and SVL 42.5���44.9 mm (N = 2) in adult females; (2) snout pointed in dorsal view; (3) tympanum moderate in size TD/ED 0.56���0.67, tubercles bearing spines on tips present on skin of temporal region including the tympanum; (4) vomerine ridge and vomerine teeth present; (5) margin of tongue rounded, not notched distally; (6) hindlimbs short, heels not meeting and tibio-tarsal articulation reaching forward to the region between tympanum and posterior corner of eye, TIB/SVL 0.37���0.43; (7) a subarticular tubercle present at the base of each fingers; (8) toes without lateral fringes and with rudiment of webbing; (9) distinct sparse enlarged tubercles on the surface of limbs, flanks, posterior part of belly and around the cloaca; (10) tips of the tubercles on ventral surface of thighs and around the cloaca bearing tiny spines; (11) single subgular vocal sac in males; (12) nuptial pads with well-developed villiform black nuptial spines on the dorsal surface of the first and second fingers in breeding males. Comparisons. Comparative data of Boulenophrys fengshunensis sp. nov. and the other recognized members of Boulenophrys are listed in Table 3. Boulenophrys fengshunensis sp. nov. can be distinguished from Boulenophrys puningensis sp. nov. and Boulenophrys hungtai sp. nov. having a tympanum with tubercles bearing spines on their tips (vs. tympanic region smooth without granules, tubercles or spines in Boulenophrys puningensis sp. nov. and Boulenophrys hungtai sp. nov.); the presence of both a vomerine ridge and vomerine teeth (vs. absence of vomerine ridge but with vomerine teeth present in Boulenophrys hungtai sp. nov.); tubercles on skin of the posterior part of belly without spines (vs. tubercles on skin of the posterior part of belly bearing spines on their tips in both Boulenophrys puningensis sp. nov. and Boulenophrys hungtai sp. nov.). With a smaller body size, SVL 34.3���39.4 mm in adult males and SVL 42.5���44.9 mm in adult females, Boulenophrys fengshunensis sp. nov. differs from the seven congeners whose SVL ��� 50 mm in males or SVL ��� 60 mm in females, including B. caudoprocta (81.3 mm in a single male), B. jingdongensis (53.0��� 56.5 mm in males and 63.5 mm in a single female), B. liboensis (60.5���67.7 mm in males and 60.8���70.6 in females), B. mirabilis (55.8���61.4 mm in males and 68.5���74.8 mm in females), B. omeimontis (56.0��� 59.5 mm in males and 68.0��� 72.5 mm in females), B. sangzhiensis (54.7 mm in a single male), and B. shuichengensis (102.0��� 118.3 mm in males and 99.8���115.6 mm in females). In having relatively shorter shanks with heels that do not meet when the flexed hind limbs are held at right angles to the body axis, Boulenophrys fengshunensis sp. nov. can be easily distinguished from the following 35 congeners, i.e. B. angka, B. anlongensis, B. baishanzuensis, B. baolongensis, B. binchuanensis, B. binlingensis, B. boettgeri, B. congjiangensis, B. cheni, B. chishuiensis, B. daiyunensis, B. jiangi, B. jinggangensis, B. jiulianensis, B. leishanensis, B. lini, B. minor, B. mufumontana, B. nanlingensis, B. palpebralespinosa, B. qianbeiensis, B. sanmingensis, B. shimentaina, B. shunhuangensis, B. spinata, B. tongboensis, B. tuberogranulatus, B. wuliangshanensis, B. wushanensis, B. xiangnanensis, B. xianjuensis, B. yaoshanensis, B.yangmingensis, B. yingdeensis and B. yunkaiensis, all of which have relatively longer shanks with the heels meeting or overlapping. By the presence of vomerine teeth, Boulenophrys fengshunensis sp. nov. differs from B. acuta, B. brachykolos, B. caobangensis, B. daoji, B. huangshanensis, B. kuatunensis, B. lishuiensis, B. lushuiensis, B. obesa, B. ombrophila, and B. wugongensis, all of which lack vomerine teeth. By having a rounded margin of the tongue that is not notched distally, Boulenophrys fengshunensis sp. nov. differs from B. hoanglienensis, B. insularis, which have a notched tongue. By the absence of lateral fringes on toes, Boulenophrys fengshunensis sp. nov. differs from B. rubrimera, which has narrow lateral fringes on toes. By the presence of rudimentary webbing on toes, Boulenophrys fengshunensis sp. nov. differs from B. daweimontis, B. fansipanensis, B. frigida, and B. rubrimera, all of which lack webbing on toes. Boulenophrys fengshunensi sp. nov. further differs from the remaining B. dongguanensis and B. nankunensis by having raised tubercles on the surface of posterior abdomen (vs. absence of such tubercles in both B. dongguanensis B. nankunensis); the presence of tubercles bearing spines on their tips on the temporal region including the tympanum (vs. temporal region lacking tubercles or spines in both B. dongguanensis and B. nankunensis). Description of holotype. Adult male. Small body size, SVL 37.4 mm; head width slightly larger than head length, HWD/HDL 1.17; snout pointed in dorsal view, projecting, sloping backward to mouth in profile, protruding well beyond margin of lower jaw; top of head flat; eyes moderate in size, ED 0.37 of HDL, pupil vertical, near diamond-shaped; nostril oblique-ovoid; canthus rostralis well developed; loreal region slightly oblique; internasal distance slightly larger than interorbital distance; tympanum moderate in size with an obvious margin, TD/ED 0.65; large ovoid choanae at base of maxilla; vomerine ridge and vomerine teeth present, maxillary teeth present; margin of tongue rounded, not notched distally; presence of single subgular vocal sac, and pair of slit-like openings at posterior of jaw. Forearm (i.e., radioulna) length 0.21 of SVL and hand 0.25 of SVL; hand without webbing, fingers without lateral fringes, relative finger length I = II Coloration of holotype in life. Dorsal surface of body dark brown, with incomplete dark brown triangular marking between eyes. Two wide oblique black bands present on forearm. Dorsal surface of fingers and hindlimbs with dark grey transverse bands. Presence of vertical dark brown band below eye. Tip of tubercles on edge of upper eyelid greyish white. Supratympanic fold orangey brown. Ventral surface dark brown, black longitudinal band on surface of throat, posterior part of belly with irregular greyish white patches. Tubercles on ventral surface of posterior part of belly and thighs greyish white; spines on tips of tubercles on temporal region, ventral surface of thighs and around cloaca dark grey. Digits, inner and outer metacarpal tubercles, and inner metatarsal tubercle dark grey. Pectoral glands beige and femoral glands greyish white. Iris orangish brown. Coloration of holotype in preservative. Dorsal surface of body dark brown. Triangular marking between eyes, vertical dark brown band below eye and transverse bands on dorsal forearms and hind limbs become indistinct. Supratympanic fold greyish white. All bands and patterns on ventral surface no longer apparent. Irregular greyish white patches on posterior part of belly become dark grey. Variation. Mensural data of the type series are listed in Table 6. Most of the paratypes are similar to the holotype in morphology and color pattern, except for the following: dorsal surface and ventral surface dark brown, posterior part of belly with irregular greyish white patches, ventral surface of thighs dark grey, triangular marking between eyes is incomplete in the holotype (vs. dorsal surface beige and ventral surface light orange, posterior part of belly and ventral surface of thighs greyish white, triangular marking between eyes is complete in the female paratype SYS a005221 (Fig. 8); tibio-tarsal articulation reaching forward to middle of tympanum when hind limb stretched along body in the holotype (vs. tibio-tarsal articulation reaching forward to posterior margin of tympanum in the paratype SYS a004725; reaching forward to the region between tympanum and eye in the paratype SYS a004726). Females are distinctly larger than the males. Distribution and natural history. Currently, Boulenophrys fengshunensis sp. nov. is only known from its type locality, Mt. Tongguzhang of Fengshun. It inhabits flowing montane streams and the nearby forest floor and leaf litter at elevations between 800���1500 m. Boulenophrys fengshunensis sp. nov. is found to be sympatric with Pachytriton granulosus Chang, 1933 and Cynops glaucus Yuan, Jiang, Ding, Zhang & Che, 2013. Advertisement calls of males were heard during April and June. Males were found calling under the leaf litters around the flowing seeps. Tadpoles could be found in this period. Female specimens collected during August had immature eggs; males were not heard calling during this period, but sub-adults were observed., Published as part of Wang, Jian, Zeng, Zhao-Chi, Lyu, Zhi-Tong, Qi, Shuo, Liu, Zu-Yao, Chen, Hong-Hui, Lu, Yu-Hong, Xiao, Hui-Wen, Lin, Can-Rong, Chen, Kai & Wang, Ying-Yong, 2022, Description of three new Boulenophrys species from eastern Guangdong, China, emphasizing the urgency of ecological conservation in this region (Anura, Megophryidae), pp. 91-119 in Zootaxa 5099 (1) on pages 110-114, DOI: 10.11646/zootaxa.5099.1.4, http://zenodo.org/record/6036964, {"references":["Liu, Z. Y., Chen, G. L., Zhu, T. Q., Zeng, Z. C., Lyu, Z. T., Wang, J., Messenger, K., Greenberg, A. J., Guo, Z. X., Yang, Z. H., Shi, S. H. & Wang, Y. Y. (2018) Prevalence of cryptic species in morphologically uniform taxa - Fast speciation and evolutionary radiation in Asian frogs. Molecular Phylogenetics and Evolution, 127, 723 - 731. https: // doi. org / 10.1016 / j. ympev. 2018.06.020","Chang, M. L. Y. (1933) On the salamanders of Chekiang. Contributions from the Biological Laboratory of the Science Society of China. Zoological Series, 9, 305 - 328.","Yuan, Z. Y., Jiang, K., Ding, L., Zhang, L. & Che, J. (2013) A new newt of the genus Cynops (Caudata: Salamandridae) from Guangdong, China. Asian Herpetological Research, 4 (2), 116 - 123. https: // doi. org / 10.3724 / SP. J. 1245.2013.00116"]}

Boulenophrys puningensis sp. nov. Wang, Zeng, Lyu, Xiao & Wang Puning Horned Toad (in English) / Pǔ Níng Jiǎo Chán (DṪDzdz in Chinese) Figures 3–4 Holotype. SYS a005770, adult male, collected by Jian Wang on 24April 2017 from Longkeng Village (23°7'54.07"N, 115°51'5.28"E; ca. 120 m a.s.l.), Daping Town, Puning, Jieyang, Guangdong, China. Paratypes (N=5). Adult male, SYS a006755/ CIB118526, collected by Jian Wang, Can-Rong Lin and Hui-Wen Xiao on 14 February 2018; adult males SYS a007649, 7650 and adult females SYS a007647, 7648, collected by Jian Wang, Can-Rong Lin and Hui-Wen Xiao on 18 March 2019, all from the same stream as the holotype at elevations between 250–300 m. Etymology. The specific epithet “ puningensis ” refers to the type locality of the new species in Puning. Three of the authors of this work (Jian Wang, Hui-Wen Xiao and Can-Rong Lin) chose this nomen in honor of their hometown. Diagnosis. (1) Small body size, SVL 31.7–34.6 mm (33.0 ± 1.3, N = 4) in adult males and SVL 37.8–38.3 mm (N = 2) in adult females; (2) snout rounded in dorsal view; (3) tympanum large, TD/ED 0.68–0.71; (4) tympanic region smooth without granules or tubercles; (5) vomerine ridge and vomerine teeth present; (6) margin of tongue rounded, not notched distally; (7) hindlimbs short, heels not meeting and tibio-tarsal articulation reaching forward to the region between tympanum and eye; (8) a subarticular tubercle present at the base of each fingers; (9) toes without lateral fringes and with rudiment of webbing; (10) distinct enlarged tubercles on the surface of limbs, flanks, chest, belly and around the cloaca; (11) tips of the enlarged tubercles on posterior abdomen, ventral thighs and around the cloaca bearing tiny spines; (12) single subgular vocal sac in males; (13) nuptial pads with villiform black nuptial spines on the dorsal surface of the first and second fingers in breeding males. Comparisons. Comparative data of Boulenophrys puningensis sp. nov. and the other recognized members of Boulenophrys are listed in Table 3. With a smaller body size, SVL 31.7–34.6 mm in adult males and SVL 37.8–38.3 mm in adult females, Boulenophrys puningensis sp. nov. differs from the eight congeners whose SVL ≥ 50 mm in adult males or females, including B. caudoprocta (81.3 mm in a single male), B. jingdongensis (53.0– 56.5 mm in males and 63.5 mm in a single female), B. liboensis (60.5–67.7 mm in males and 60.8–70.6 in females), B. mirabilis (55.8–61.4 mm in males and 68.5–74.8 mm in females), B. omeimontis (56.0– 59.5 mm in males and 68.0– 72.5 mm in females), B. sangzhiensis (54.7 mm in a single male), B. shuichengensis (102.0– 118.3 mm in males and 99.8–115.6 mm in females), and B. spinata (54.0–55.0 mm in females). Boulenophrys puningensis sp. nov. shows the least genetic divergence from B. kuatunensis (mean p -distances 5.3 % in the 16S gene) and B. daiyunensis (mean p -distances 6.2 % in the 16S gene). However, the new species distinctively differs from these species by having relatively shorter shanks with the heels not meeting when the flexed hindlimbs are held at right angles to the body axis (vs. heels meeting or overlapping in B. daiyunensis); having rudiment of webbing and no lateral fringes on toes (vs. lateral fringes narrow in B. daiyunensis; webbing absent in B. kuatunensis); having raised and enlarged tubercles with spines on their tips on surface of posterior abdomen, ventral thighs and around the cloaca (vs. such tubercles not enlarged and without spines in B. daiyunensis; ventral surface smooth in B. kuatunensis). Boulenophrys puningensis sp. nov. is morphologically most similar to B. brachykolos, which is restricted to Hong Kong and Shenzhen, China (Liu et al. 2018). The new species differs from B. brachykolos by having vomerine teeth (vs. absent in B. brachykolos); lacking spines on the surface of the tympanic region (vs. having dense tiny spines on the surface of the tympanic region in B. brachykolos); and having different relative finger length formula (I = II Banophrys puningensis sp. nov. vs. II B. brachykolos). In having relatively shorter shanks with heels that do not meet when the flexed hind limbs are held at right angles to the body axis, Boulenophrys puningensis sp. nov. can be easily distinguished from the following 32 congeners, i.e. B. angka, B. anlongensis, B. baishanzuensis, B. baolongensis, B. binchuanensis, B. binlingensis, B. boettgeri, B. congjiangensis, B. cheni, B. chishuiensis, B. jiangi, B. jinggangensis, B. jiulianensis, B. leishanensis, B. lini, B. minor, B. mufumontana, B. nanlingensis, B. palpebralespinosa, B. qianbeiensis, B. sanmingensis, B. shimentaina, B. shunhuangensis, B. tongboensis, B. wuliangshanensis, B. wushanensis, B. xiangnanensis, B. xianjuensis, B. yaoshanensis, B. yangmingensis, B. yingdeensis and B. yunkaiensis, all of which have relatively longer shanks with the heels meeting or overlapping. By the presence of vomerine teeth, Boulenophrys puningensis sp. nov. differs from B. acuta, B. caobangensis, B. daoji, B. huangshanensis, B. lishuiensis, B. lushuiensis, B. obesa, B. ombrophila, B. tuberogranulatus, and B. wugongensis, all of which lack vomerine teeth. By having a rounded tongue margin that is not notched distally, Boulenophrys puningensis sp. nov. differs from B. hoanglienensis, and B. insularis, all of which have notched tongues. By the absence of lateral fringes on toes, Boulenophrys puningensis sp. nov. differs from B. rubrimera, which has narrow lateral fringes on toes. By the presence of rudimentary webbing on the toes, Boulenophrys puningensis sp. nov. differs from B. daweimontis, B. fansipanensis, and B. frigida, all of which lack webbing on the toes. Boulenophrys puningensis sp. nov. further differs from the remaining B. dongguanensis and B. nankunensis by having raised tubercles bearing spines on the surface of the posterior abdomen, ventral thighs, and around the cloaca (vs. absence of such tubercles and spines in B. dongguanensis and B. nankunensis). Description of holotype. Adult male. Body size small, SVL 34.6 mm. Head width slightly larger than head length, HWD/HDL 1.03; snout rounded in dorsal view, projecting, sloping backward to mouth in profile, protruding well beyond margin of lower jaw; top of head flat; eyes moderate in size, ED 0.36 of HDL, pupil vertical, near diamond-shaped; nostril oblique-ovoid; canthus rostralis well developed; loreal region slightly oblique; internasal distance slightly larger than interorbital distance; tympanum large with an obvious margin, TD/ED 0.70; large ovoid choanae at base of maxilla; vomerine ridge and vomerine teeth present, maxillary teeth present; margin of tongue rounded, not notched distally; presence of single subgular vocal sac, and pair of slit-like openings at posterior of jaw. Forearm (i.e., radioulna) length 0.21 of SVL and hand 0.23 of SVL; hand without webbing, fingers without lateral fringes, relative finger length I = II Dorsal skin rough and granular, with raised conical tubercles; sparse large tubercles on flanks; single horn-like prominent tubercle on edge of upper eyelid; obvious supratympanic fold curving posteroventrally from posterior corner of eye to level above insertion of arm; tympanic region smooth without granules or tubercles; dense tubercles on skin of upper lip, upper eyelid, mandibular articulation, loreal, temporal region excluding tympanum and surface around cloaca; a single discontinuous “V” shaped ridge present on occipital region; dense tubercles on shanks and thighs; ventral surface with dense raised tubercles; tubercles on surface of posterior abdomen, ventral surface of thighs and around cloaca bearing tiny spines on their tips; small pectoral gland closer to axilla; single femoral gland positioned on posterior surface of thigh at midpoint between knee and cloaca. Coloration of holotype in life. Dorsal surface of body yellowish brown, with incomplete dark brown triangular marking between eyes. Two wide oblique black bands present on forearm. Dorsal surface of fingers and hindlimbs with dark grey transverse bands. Presence of vertical dark brown band below eye. Tubercles on edge of upper eyelid beige. Supratympanic fold light brown. Ventral surface dark grey, with black longitudinal band on surface of throat; surface of throat and chest mottled with orange patches. Tubercles on ventral surface of chest, belly, and thighs greyish white; spines on tips of tubercles on surface of posterior abdomen; ventral surface of thighs and around cloaca dark grey. Digits, inner and outer metacarpal tubercles and inner metatarsal tubercle greyish white. Pectoral glands and femoral glands beige, mottled with orange patches. Iris yellowish brown, with greyish white patches on upper and lower margin. Coloration of holotype in preservative. Yellowish brown fades to greyish brown dorsally. Color of the triangular marking between eyes, oblique bands on forearms, patterns on ventral surface faded. Orange patches on surface of throat, chest; color of pectoral glands and femoral glands faded. Variation. Mensural data of the type series are listed in Table 4. Most of the paratypes are similar to the holotype in morphology and color pattern, except for the following: dorsal surface of body yellowish brown in the holotype (vs. dorsal surface of body light brown in the paratypes SYS a007647 (Fig. 4C) and SYS a007648 (Fig. 4E); ventral surface dark grey with orange patches (vs. ventral surface lacking bright patches in the paratypes SYS a007649 (Fig. 4B), SYS a007647 (Fig. 4D) and SYS a007648 (Fig. 4F); iris yellowish brown, with greyish white patches on its upper and lower margin in the holotype (vs. iris grey with beige and dark mottling in the paratype SYS a007649 (Fig. 4A); tubercles on posterior part of abdomen of the paratype SYS a007648 (Fig. 4F) are weakly developed. Females are distinctly larger than the males. Distribution and natural history. Currently, Boulenophrys puningensis sp. nov. is only known from its type locality, Longkeng Village of Puning. It inhabits flowing montane streams and the nearby forest floor and leaf litter at elevations between 120– 300 m. Advertisement calls of males were heard from February until April. Males were found calling in rock crevices in the flowing streams. Tadpoles could be found in this period.

Boulenophrys puningensis sp. nov. Wang, Zeng, Lyu, Xiao & Wang Puning Horned Toad (in English) / P�� N��ng Ji��o Ch��n (D���Dzdz in Chinese) Figures 3���4 Holotype. SYS a005770, adult male, collected by Jian Wang on 24April 2017 from Longkeng Village (23��7'54.07"N, 115��51'5.28"E; ca. 120 m a.s.l.), Daping Town, Puning, Jieyang, Guangdong, China. Paratypes (N=5). Adult male, SYS a006755/ CIB118526, collected by Jian Wang, Can-Rong Lin and Hui-Wen Xiao on 14 February 2018; adult males SYS a007649, 7650 and adult females SYS a007647, 7648, collected by Jian Wang, Can-Rong Lin and Hui-Wen Xiao on 18 March 2019, all from the same stream as the holotype at elevations between 250���300 m. Etymology. The specific epithet ��� puningensis ��� refers to the type locality of the new species in Puning. Three of the authors of this work (Jian Wang, Hui-Wen Xiao and Can-Rong Lin) chose this nomen in honor of their hometown. Diagnosis. (1) Small body size, SVL 31.7���34.6 mm (33.0 �� 1.3, N = 4) in adult males and SVL 37.8���38.3 mm (N = 2) in adult females; (2) snout rounded in dorsal view; (3) tympanum large, TD/ED 0.68���0.71; (4) tympanic region smooth without granules or tubercles; (5) vomerine ridge and vomerine teeth present; (6) margin of tongue rounded, not notched distally; (7) hindlimbs short, heels not meeting and tibio-tarsal articulation reaching forward to the region between tympanum and eye; (8) a subarticular tubercle present at the base of each fingers; (9) toes without lateral fringes and with rudiment of webbing; (10) distinct enlarged tubercles on the surface of limbs, flanks, chest, belly and around the cloaca; (11) tips of the enlarged tubercles on posterior abdomen, ventral thighs and around the cloaca bearing tiny spines; (12) single subgular vocal sac in males; (13) nuptial pads with villiform black nuptial spines on the dorsal surface of the first and second fingers in breeding males. Comparisons. Comparative data of Boulenophrys puningensis sp. nov. and the other recognized members of Boulenophrys are listed in Table 3. With a smaller body size, SVL 31.7���34.6 mm in adult males and SVL 37.8���38.3 mm in adult females, Boulenophrys puningensis sp. nov. differs from the eight congeners whose SVL ��� 50 mm in adult males or females, including B. caudoprocta (81.3 mm in a single male), B. jingdongensis (53.0��� 56.5 mm in males and 63.5 mm in a single female), B. liboensis (60.5���67.7 mm in males and 60.8���70.6 in females), B. mirabilis (55.8���61.4 mm in males and 68.5���74.8 mm in females), B. omeimontis (56.0��� 59.5 mm in males and 68.0��� 72.5 mm in females), B. sangzhiensis (54.7 mm in a single male), B. shuichengensis (102.0��� 118.3 mm in males and 99.8���115.6 mm in females), and B. spinata (54.0���55.0 mm in females). Boulenophrys puningensis sp. nov. shows the least genetic divergence from B. kuatunensis (mean p -distances 5.3 % in the 16S gene) and B. daiyunensis (mean p -distances 6.2 % in the 16S gene). However, the new species distinctively differs from these species by having relatively shorter shanks with the heels not meeting when the flexed hindlimbs are held at right angles to the body axis (vs. heels meeting or overlapping in B. daiyunensis); having rudiment of webbing and no lateral fringes on toes (vs. lateral fringes narrow in B. daiyunensis; webbing absent in B. kuatunensis); having raised and enlarged tubercles with spines on their tips on surface of posterior abdomen, ventral thighs and around the cloaca (vs. such tubercles not enlarged and without spines in B. daiyunensis; ventral surface smooth in B. kuatunensis). Boulenophrys puningensis sp. nov. is morphologically most similar to B. brachykolos, which is restricted to Hong Kong and Shenzhen, China (Liu et al. 2018). The new species differs from B. brachykolos by having vomerine teeth (vs. absent in B. brachykolos); lacking spines on the surface of the tympanic region (vs. having dense tiny spines on the surface of the tympanic region in B. brachykolos); and having different relative finger length formula (I = II Banophrys puningensis sp. nov. vs. II B. brachykolos). In having relatively shorter shanks with heels that do not meet when the flexed hind limbs are held at right angles to the body axis, Boulenophrys puningensis sp. nov. can be easily distinguished from the following 32 congeners, i.e. B. angka, B. anlongensis, B. baishanzuensis, B. baolongensis, B. binchuanensis, B. binlingensis, B. boettgeri, B. congjiangensis, B. cheni, B. chishuiensis, B. jiangi, B. jinggangensis, B. jiulianensis, B. leishanensis, B. lini, B. minor, B. mufumontana, B. nanlingensis, B. palpebralespinosa, B. qianbeiensis, B. sanmingensis, B. shimentaina, B. shunhuangensis, B. tongboensis, B. wuliangshanensis, B. wushanensis, B. xiangnanensis, B. xianjuensis, B. yaoshanensis, B. yangmingensis, B. yingdeensis and B. yunkaiensis, all of which have relatively longer shanks with the heels meeting or overlapping. By the presence of vomerine teeth, Boulenophrys puningensis sp. nov. differs from B. acuta, B. caobangensis, B. daoji, B. huangshanensis, B. lishuiensis, B. lushuiensis, B. obesa, B. ombrophila, B. tuberogranulatus, and B. wugongensis, all of which lack vomerine teeth. By having a rounded tongue margin that is not notched distally, Boulenophrys puningensis sp. nov. differs from B. hoanglienensis, and B. insularis, all of which have notched tongues. By the absence of lateral fringes on toes, Boulenophrys puningensis sp. nov. differs from B. rubrimera, which has narrow lateral fringes on toes. By the presence of rudimentary webbing on the toes, Boulenophrys puningensis sp. nov. differs from B. daweimontis, B. fansipanensis, and B. frigida, all of which lack webbing on the toes. Boulenophrys puningensis sp. nov. further differs from the remaining B. dongguanensis and B. nankunensis by having raised tubercles bearing spines on the surface of the posterior abdomen, ventral thighs, and around the cloaca (vs. absence of such tubercles and spines in B. dongguanensis and B. nankunensis). Description of holotype. Adult male. Body size small, SVL 34.6 mm. Head width slightly larger than head length, HWD/HDL 1.03; snout rounded in dorsal view, projecting, sloping backward to mouth in profile, protruding well beyond margin of lower jaw; top of head flat; eyes moderate in size, ED 0.36 of HDL, pupil vertical, near diamond-shaped; nostril oblique-ovoid; canthus rostralis well developed; loreal region slightly oblique; internasal distance slightly larger than interorbital distance; tympanum large with an obvious margin, TD/ED 0.70; large ovoid choanae at base of maxilla; vomerine ridge and vomerine teeth present, maxillary teeth present; margin of tongue rounded, not notched distally; presence of single subgular vocal sac, and pair of slit-like openings at posterior of jaw. Forearm (i.e., radioulna) length 0.21 of SVL and hand 0.23 of SVL; hand without webbing, fingers without lateral fringes, relative finger length I = II Coloration of holotype in life. Dorsal surface of body yellowish brown, with incomplete dark brown triangular marking between eyes. Two wide oblique black bands present on forearm. Dorsal surface of fingers and hindlimbs with dark grey transverse bands. Presence of vertical dark brown band below eye. Tubercles on edge of upper eyelid beige. Supratympanic fold light brown. Ventral surface dark grey, with black longitudinal band on surface of throat; surface of throat and chest mottled with orange patches. Tubercles on ventral surface of chest, belly, and thighs greyish white; spines on tips of tubercles on surface of posterior abdomen; ventral surface of thighs and around cloaca dark grey. Digits, inner and outer metacarpal tubercles and inner metatarsal tubercle greyish white. Pectoral glands and femoral glands beige, mottled with orange patches. Iris yellowish brown, with greyish white patches on upper and lower margin. Coloration of holotype in preservative. Yellowish brown fades to greyish brown dorsally. Color of the triangular marking between eyes, oblique bands on forearms, patterns on ventral surface faded. Orange patches on surface of throat, chest; color of pectoral glands and femoral glands faded. Variation. Mensural data of the type series are listed in Table 4. Most of the paratypes are similar to the holotype in morphology and color pattern, except for the following: dorsal surface of body yellowish brown in the holotype (vs. dorsal surface of body light brown in the paratypes SYS a007647 (Fig. 4C) and SYS a007648 (Fig. 4E); ventral surface dark grey with orange patches (vs. ventral surface lacking bright patches in the paratypes SYS a007649 (Fig. 4B), SYS a007647 (Fig. 4D) and SYS a007648 (Fig. 4F); iris yellowish brown, with greyish white patches on its upper and lower margin in the holotype (vs. iris grey with beige and dark mottling in the paratype SYS a007649 (Fig. 4A); tubercles on posterior part of abdomen of the paratype SYS a007648 (Fig. 4F) are weakly developed. Females are distinctly larger than the males. Distribution and natural history. Currently, Boulenophrys puningensis sp. nov. is only known from its type locality, Longkeng Village of Puning. It inhabits flowing montane streams and the nearby forest floor and leaf litter at elevations between 120��� 300 m. Advertisement calls of males were heard from February until April. Males were found calling in rock crevices in the flowing streams. Tadpoles could be found in this period., Published as part of Wang, Jian, Zeng, Zhao-Chi, Lyu, Zhi-Tong, Qi, Shuo, Liu, Zu-Yao, Chen, Hong-Hui, Lu, Yu-Hong, Xiao, Hui-Wen, Lin, Can-Rong, Chen, Kai & Wang, Ying-Yong, 2022, Description of three new Boulenophrys species from eastern Guangdong, China, emphasizing the urgency of ecological conservation in this region (Anura, Megophryidae), pp. 91-119 in Zootaxa 5099 (1) on pages 102-106, DOI: 10.11646/zootaxa.5099.1.4, http://zenodo.org/record/6036964, {"references":["Liu, Z. Y., Chen, G. L., Zhu, T. Q., Zeng, Z. C., Lyu, Z. T., Wang, J., Messenger, K., Greenberg, A. J., Guo, Z. X., Yang, Z. H., Shi, S. H. & Wang, Y. Y. (2018) Prevalence of cryptic species in morphologically uniform taxa - Fast speciation and evolutionary radiation in Asian frogs. Molecular Phylogenetics and Evolution, 127, 723 - 731. https: // doi. org / 10.1016 / j. ympev. 2018.06.020"]}

Acute myeloid leukemia (AML) with nucleophosmin 1 (NPM1) mutations exhibits distinct biological and clinical features, accounting for approximately one-third of AML. Recently, the N6-methyladenosine (m6A) RNA modification has emerged as a new epigenetic modification to contribute to tumorigenesis and development. However, there is limited knowledge on the role of m6A modifications in NPM1-mutated AML. In this study, the decreased m6A level was first detected and high expression of fat mass and obesity-associated protein (FTO) was responsible for the m6A suppression in NPM1-mutated AML. FTO upregulation was partially induced by NPM1 mutation type A (NPM1-mA) through impeding the proteasome pathway. Importantly, FTO promoted leukemic cell survival by facilitating cell cycle and inhibiting cell apoptosis. Mechanistic investigations demonstrated that FTO depended on its m6A RNA demethylase activity to activate PDGFRB/ERK signaling axis. Our findings indicate that FTO-mediated m6A demethylation plays an oncogenic role in NPM1-mutated AML and provide a new layer of epigenetic insight for future treatments of this distinctly leukemic entity.

Background: Hyaluronan mediated motility receptor (also known as RHAMM) is another one of few defined hyaluronan receptors, play pivotal roles in cell growth. However, the functionof HMMR in lung adenocarcinoma remain unclear.Methods: HMMR expression was analyzed emoloyed the public databases, the prognosis of HMMR was analysis by prognoscan, KMplot and GEPIA databases. The GO and KEGG pathway was analysis by the DAVID and GSEA software. The correlation between the HMMR expression was analysis by the TIMER databases, the gene and protein networks was analysis by Genemania and STRING databases, the DNA methylation was analysis by the MethSurv and UALCAN databases. The expression of HMMR was analysis by IHC and qPCR, the function of HMMR on cell proliferation and migration was examine by the cell growth curve, clone information, transwell and wound healing assay.Results: in this study, we find that HMMR was elevated in LUAD and it’s highly expression associated with the poor prognosis and lymph node metastasis. Furthermore, the expression of HMMR was induced by hypoxia in LUAD. HMMR expression level not only positively correlation with the different immune cells, but also positively correlation with the expression of immune checkpoints related gene. Finally, depletion of HMMR significantly represses the cell growth and migration of NSCLC. We also found that the HOXB7/TMPO-AS1/Let-7b-5p axis mediated high expression of HMMR in NSCLC, depletion of TMPO-AS1 and over-expression the Let-7b-5p would result in decreased the expression of HMMR in NSCLC cells, the TMPO-AS1 was positively with the HMMR and negatively related to the Let-7b-5p in NSCLC. Overall, this study emphasized the significance of HOXB7/TMPO-AS1/Let-7b-5p axis mediated high expression of HMMR in cancer progression and Immune infiltration of LUAD.Conclusions: we demonstrated HMMR was elevated in LUAD and positively relation to poor prognosis. We find the hypoxia microenvironment and DNA hypomethylation able to up-regulation of the HMMR expression. Additionally, HMMR expression was positive with the diverse immune cell and immune regulator related gene in LUAD. Finally, we found that depletion of HMMR was inhibits the cell proliferation and migration ability of NSCLC cells. These findings suggest that HMMR could be served as a biomarker for prognosis and immune infiltration in LUAD.

BackgroundHyaluronan mediated motility receptor (also known as RHAMM) is another one of few defined hyaluronan receptors, play pivotal roles in cell growth. However, the relationships between HMMR and prognosis and tumor-infiltrating lymphocytes in lung adenocarcinoma remain unclear.MethodsHMMR expression was analyzed emoloyed the TIMER, GEPIA, UALCAN, CCLE databases, the prognosis of HMMR was analysis by prognoscan, KMplot and GEPIA databases. The GO and KEGG pathway was analysis by the DAVID and GSEA software. The correlation between the HMMR expression was analysis by the TIMER databases, the gene and protein networks was analysis by Genemania and STRING databases, the DNA methylation was analysis by the MethSurv and UALCAN databases, the gene mutation of HMMR was analysis by the cBioportal and COSMIC databases. The expression of HMMR was analysis by IHC and qPCR, the function of HMMR on cell proliferation and migration was examine by the cell growth curve, clone information, transwell and wound healing assay.ResultsIn this study, we find that HMMR was elevated in LUAD and it’s highly expression associated with the poor prognosis and lymph node metastasis. Furthermore, the expression of HMMR was induced by hypoxia in LUAD. HMMR expression level not only positively correlation with the different immune cells, but also positively correlation with the expression of immune checkpoints related gene, for instance, CD279, CD274, CTLA4, LAG3, PDCD1LG2, TIGIT and HAVCR2. Finally, depletion of HMMR significantly represses the cell growth and migration of NSCLC. Overall, this study emphasized the significance of HMMR in cancer progression and Immune infiltration of LUAD.ConclusionsWe demonstrated HMMR was elevated in LUAD and positively relation to poor prognosis. We find the hypoxia microenvironment and DNA hypomethylation able to up-regulation of the HMMR expression. Additionally, HMMR expression was positive with the diverse immune cell and immune regulator related gene in LUAD. Finally, we found that depletion of HMMR was inhibits the cell proliferation and migration ability of NSCLC cells. These findings suggest that HMMR could be served as a biomarker for prognosis and immune infiltration in LUAD.

Rui-huan Gan,1,* Li-song Lin,2,* Jing Xie,3–5,* Li Huang,2,4 Lin-can Ding,2 Bo-hua Su,2 Xian-e Peng,1,5 Da-li Zheng,4,5 You-guang Lu1,3 1Department of Epidemiology and Health Statistics, Fujian Provincial Key Laboratory of Environment Factors and Cancer, School of Public Health, Fujian Medical University, Fuzhou 350122, People’s Republic of China; 2Department of Oral and Maxillofacial Surgery, Affiliated First Hospital of Fujian Medical University, Fuzhou 350005, People’s Republic of China; 3Department of Preventive Dentistry, School and Hospital of Stomatology, Fujian Medical University, Fuzhou 350000, People’s Republic of China; 4Key Laboratory of Stomatology of Fujian Province, School and Hospital of Stomatology, Fujian Medical University, Fuzhou 350004, People’s Republic of China; 5Key Laboratory of Ministry of Education for Gastrointestinal Cancer, Fujian Medical University, Fuzhou 350122, People’s Republic of China*These authors contributed equally to this workCorrespondence: You-guang LuDepartment of Preventive Dentistry, School and Hospital of Stomatology, Fujian Medical University, 246 Yang Qiao Middle Road, Fuzhou 350000, People’s Republic of ChinaTel +86 591 8373 6429Fax +86 591 8372 0599Email fjlyg63@fjmu.edu.cnXian-e PengDepartment of Epidemiology and Health Statistics, Fujian Provincial Key Laboratory of Environment Factors and Cancer, School of Public Health, Fujian Medical University, 1 Xue Yuan Road, University Town, Fuzhou 350122, People’s Republic of ChinaTel +86 138 0951 6580Fax +86 591 8372 0599Email fmuxe@163.comPurpose: The Notch signaling pathway plays an oncogenic role in tongue squamous cell carcinoma. The aim of this study was to inhibit the proliferation and self-renewal of tongue cancer cells by applying Notch signaling pathway inhibitor FLI-06 (Selleck, USA) and to lay a foundation for the clinically targeted treatment of tongue cancer for the future.Methods: The mRNA expression level of Notch1 and the overall survival rate of patients with tongue cancer were examined by analyzing the TCGA database. Tongue cancer cells were treated with FLI-06. Cell proliferation, apoptosis, and stem cell self-renewal ability were tested in appropriate ways. A xenograft mouse model was established to observe tumor growth.Results: From the TCGA data, we demonstrated that patients with high expression of Notch1 had a poor prognosis. We observed that the Notch signaling pathway inhibitor FLI-06 can restrain the activation of the Notch signaling pathway, decrease cell proliferation and induce cell apoptosis in vitro. The xenograft experiment indicated that intraperitoneal injection of FLI-06 inhibited tumor growth and increased cell apoptosis. FLI-06 suppressed both the mRNA and protein expression of Notch receptor and Notch targeted genes. We also observed that FLI-06 suppressed the proliferation of tongue cancer stem cells.Conclusion: FLI-06 can block the proliferation and self-renewal of tongue cancer cells. It is inferred that this compound, which inhibits the Notch signaling pathway, may serve as a potential targeted drug for the treatment of tongue cancer in the clinic.Keywords: tongue cancer, Notch signaling pathway, Notch inhibitor, cancer stem cells

Purpose The Notch signaling pathway plays an oncogenic role in tongue squamous cell carcinoma. The aim of this study was to inhibit the proliferation and self-renewal of tongue cancer cells by applying Notch signaling pathway inhibitor FLI-06 (Selleck, USA) and to lay a foundation for the clinically targeted treatment of tongue cancer for the future. Methods The mRNA expression level of Notch1 and the overall survival rate of patients with tongue cancer were examined by analyzing the TCGA database. Tongue cancer cells were treated with FLI-06. Cell proliferation, apoptosis, and stem cell self-renewal ability were tested in appropriate ways. A xenograft mouse model was established to observe tumor growth. Results From the TCGA data, we demonstrated that patients with high expression of Notch1 had a poor prognosis. We observed that the Notch signaling pathway inhibitor FLI-06 can restrain the activation of the Notch signaling pathway, decrease cell proliferation and induce cell apoptosis in vitro. The xenograft experiment indicated that intraperitoneal injection of FLI-06 inhibited tumor growth and increased cell apoptosis. FLI-06 suppressed both the mRNA and protein expression of Notch receptor and Notch targeted genes. We also observed that FLI-06 suppressed the proliferation of tongue cancer stem cells. Conclusion FLI-06 can block the proliferation and self-renewal of tongue cancer cells. It is inferred that this compound, which inhibits the Notch signaling pathway, may serve as a potential targeted drug for the treatment of tongue cancer in the clinic.

El estudio expuso que los obreros municipales inicialmente eran considerados como parte de la carrera administrativa a consecuencia del Caso Huatuco, siendo esto solucionado por el TC a través de una sentencia aclaratoria para pasar a pertenecer al D.L. N°728. Sin embargo, pese a los beneficios que otorga tal decreto a favor del obrero, la Municipalidad aún los sigue contratando bajo la locación de servicios. Propuso como objetivo determinar cómo el contrato por locación de servicios vulnera los derechos fundamentales de los obreros de la Municipalidad Provincial de San Martín, 2018-2020. La investigación tuvo un enfoque mixto de tipo aplicada, nivel descriptivo y diseño correlacional - no experimental, la población se ha conformado por todos los contratos del periodo 2018-2020 y especialistas en materia laboral; la muestra por 33 contratos y 5 especialistas, utilizando como técnicas el análisis documental y entrevista, y como instrumentos la guía de análisis documental y guía de entrevista. Resultados: El contrato de locación de servicios fue desnaturalizado en los años 2018-2019, el derecho fundamental vulnerado en mayor medida fue seguridad social y existe relación entre el contrato por locación con la vulneración de los derechos fundamentales pues la significancia fue 035. Conclusión: El contrato de locación de servicios vulnera los derechos fundamentales al afectarles la seguridad social y estabilidad laboral. Recomendación: Al intendente regional de SUNAFIL-San Martín, programar y ejecutar actividades o talleres orientados a informar a las entidades e instituciones públicas de diferentes niveles de gobierno sobre el respeto de los derechos fundamentales. The study exposed that the municipal workers were initially considered as part of the administrative career as a result of the Huatuco Case, being this solved by the Constitutional Court through a clarifying sentence to become part of the D.L. N°728. However, in spite of the benefits granted by such decree in favor of the worker, the Municipality still continues to hire them under a service contract. It proposed as an objective to determine how the service contract violates the fundamental rights of the workers of the Provincial Municipality of San Martin, 2018-2020. The research had a mixed approach of applied type, descriptive level and correlational - non-experimental design, the population was made up of all the contracts of the period 2018-2020 and specialists in labor matters; the sample consisted of 33 contracts and 5 specialists, using as techniques the documentary analysis and interview, and as instruments the documentary analysis guide and interview guide. Results: The service contract was denaturalized in 2018-2019, the most violated fundamental right was social security and there is a relationship between the service contract and the violation of fundamental rights since the significance was 035. Conclusion: The service contract violates fundamental rights by affecting their social security and work stability. Recommendation: To the regional intendant of SUNAFIL-San Martín, program and implement activities or workshops aimed at informing public entities and institutions at different levels of government about respect for fundamental rights.

Shared bicycles are prevailing in China but the extent to which they contribute to maintaining and transmitting pathogens and antibiotic-resistant bacteria remain largely unknown. To fill the knowledge gap, herein, swab samples (n = 963) were collected from handlebars of shared bicycles in areas of hospital, school, metro station (n = 887) and riders (n = 76) in Chengdu, China. Staphylococci (n = 241) and Enterococci (n = 69) were widely distributed across sampling locations at a frequency of 2.3%–12.9%, and 0.08%–5.5%, respectively. Bicycle or rider-borne Gram-positive bacteria were frequently resistant to clinically important antibiotics including linezolid, fosfomycin, and vancomycin, and a significant portion of these isolates (3.4%–16.6% for Staphylococci and 0.1%–13.8% for Enterococci) indicated multidrug resistance. Nineteen Staphylococcus aureus isolates were identified in this collection and 52.6% of which were considered as methicillin-resistant S. aureus. Whole genome sequencing further characterized 26 antimicrobial resistance genes (ARGs) including fosB, fusB, and lnu(G) in S. aureus and 21 ARGs including optrA in Enterococci. Leveraging a complementary approach with conventional MLST, whole genome SNP and MLST analyses, we present that genetically closely-related bacteria were found in bicycles and riders across geographical-distinct locations suggesting bacterial transmission. Further, five new ST types 5697–5701 were firstly characterized in S. aureus. ST 942 and ST 1640 are new ST types observed in E. faecalis, and E. faecium, respectively. Our results highlighted the risk of shared bicycle system in disseminating pathogens and antibiotic resistance which warrants effective disinfections.

Previous studies have reported that inhibitor of DNA binding 1 (ID1) exerts an oncogenic role in a number of tumors. In the present study, the role of ID1 in the growth, invasion and migration of salivary adenoid cystic carcinoma (SACC) cells was investigated. ID1 expression in clinical SACC samples was compared with that in normal salivary tissues using immunohistochemical staining, and the correlation between ID1 expression and clinical pathological characteristics was then determined. Subsequently, ID1 was overexpressed or silenced to investigate the effects of ID1 expression on SACC cell proliferation, invasion and migration. In addition, the gene expression levels of known ID1 target genes, including S100A9, CDKN2A and matrix metalloproteinase 1 (MMP1) was measured using reverse transcription-quantitative polymerase chain reaction to elucidate the potential mechanisms of ID1 in SACC. The results of the present study indicated that the protein expression levels of ID1 were significantly increased in the SACC tissues compared with that in the normal salivary tissues (P

Many studies have shown that FZD2 is significantly associated with tumor development and tumor metastasis. The purpose of the present study was to gain insight into the role of FZD2 in the cell proliferation and invasion of tongue squamous cell carcinoma. According to TCGA-HNSC dataset, among the 10 Frizzled receptors, FZD2 exhibited the highest degree of differential expression between cancer tissues and normal tissues, and the overall survival of patients with higher FZD2 levels was shown to be significantly shorter compared with those with lower FZD2 levels. The upregulation of FZD2 in clinical tongue cancer tissues was validated by real-time PCR. Knockdown of FZD2 inhibited the proliferation, migration and invasion of CAL-27 and TCA-8113 cells, whereas overexpression of FZD2 led to the opposite results. Further analysis revealed that FZD2 is positively correlated with WNT3A, WNT5B, WNT7A and WNT2 and is negatively correlated with WNT4. These results indicated that FZD2 may act as an oncogene in tongue squamous cell carcinoma. Therefore, FZD2 may be a target for the diagnosis, prognosis and gene therapy of tongue cancer.

Tongue squamous cell carcinoma (TSCC) is one of the most common cancers in the oral cavity. Notch signaling is frequently dysregulated in cancer. However, the role of Notch2 in TSCC is not well understood. The aim of this study was to investigate the effect of abnormal expression of Notch2 in TSCC. The expression of Notch2 was tested in 47 pairs of tissues from tongue cancer and normal samples by using immunohistochemical staining. Tongue cancer cells were transfected with siRNA or plasmid. The proliferation of the cells was tested by the CCK8 assay and colony formation assay. Subcutaneous tumor model was established to observe tumor growth. Transwell assay was used to detect the changes of cell migration and invasion ability. A humanized anti-Notch2 antibody was used to TSCC cells. We found that Notch2 was upregulated in tongue carcinoma tissues. Knocking down the expression of Notch2 by siRNA in the TSCC cell lines decreased proliferation ability both in vitro and in vivo. In addition, migration and invasion abilities were inhibited by knockdown of Notch2 in the TSCC cells. However, overexpression of Notch2 increased tongue cancer cell proliferation, invasion and migration. The humanized anti-Notch2 antibody inhibited TSCC cell growth. The results indicated that Notch2 is an oncogene in tongue squamous cell carcinoma and may become the target of a new approach for treating TSCC.

// Jian Xie 1, * , Yan Feng 1, * , Ting Lin 1, * , Xiao-Yu Huang 1 , Rui-Huan Gan 1 , Yong Zhao 2 , Bo-Hua Su 1 , Lin-Can Ding 1 , Lin She 1 , Jiang Chen 3 , Li-Song Lin 4 , Xu Lin 5 , Da-Li Zheng 5 , You-Guang Lu 1 1 Department of Preventive Dentistry, Affiliated Stomatological Hospital, Fujian Medical University, Fuzhou, China 2 Department of Pathology, Affiliated Stomatological Hospital, Fujian Medical University, Fuzhou, China 3 Center of Dental Implant, Affiliated Stomatological Hospital, Fujian Medical University, Fuzhou, China 4 Department of Oral and Maxillofacial Surgery, Affiliated First Hospital of Fujian Medical University, Fuzhou, China 5 Key Laboratory of Ministry of Education for Gastrointestinal Cancer, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, China * These authors have contributed equally to this work Correspondence to: Da-Li Zheng, email: dalizheng@mail.fjmu.edu.cn You-Guang Lu, email: fjlyg63@163.com Keywords: CDH4, salivary adenoid cystic carcinoma, CDH1, proliferation, invasion Received: May 23, 2016 Accepted: October 14, 2016 Published: October 22, 2016 ABSTRACT The cadherin-4 gene (CDH4) of the cadherin family encodes non-epithelial R-cadherin (R-cad); however, the function of this gene in different types of cancer remains controversial. In this study, we found higher expression of CDH4 mRNA in a salivary adenoid cystic carcinoma (SACC) cell line with low metastatic potential (SACC-83) than in a cell line with high metastatic potential (SACC-LM). By analyzing 67 samples of SACC tissues and 40 samples of paraneoplastic normal tissues, we found R-cad highly expressed in 100% of normal paraneoplastic tissue but only expressed in 64% of SACC tumor tissues (P

A new system was developed for the research on the cleanings of ultrasonic acid and alkali to the used reverse osmosis membrane (UROM). By continuously detecting the absorbance, conductivity and K+/Na+/Cl−/Ca2 + contents in the cleaning solutions with this system, the cleaning information of organic and inorganic fouling on the UROM can be obtained and their cleaning effects can be judged directly. In the research, after comparing the cleaning effects of five kinds of acid solutions to the used polyamide ROMs, it was found that the ultrasonic cleaning effect of oxalic acid is the best. Therefore the concentration and pH of the oxalic acid, the sequence of acid and alkali cleanings were investigated, the results showed that, when the pH of 0.2% oxalic acid was regulated to 2.5 by the ammonia water, the defouling rate of the UROM can be reached 91%, and if employing the acid and alkali solutions to simultaneously clean, the best cleaning effect will be obtained, in which the cleaning order should be the first acid then alkali plus the ultrasound. The results and parameters obtained in this study can be used as a reference basis for designing large-scale chemical and ultrasonic cleaning system of UROM.

For a control box with concentrated power consumption, high heat generation, and small volume requirements, the paper designed a new piezoelectric fan cooling system. The system used piezoelectric ceramics as the fan blade drive module, could achieve directional, efficient and reliable heat dissipation, ensured stable and safe operation of key components. The composition and characteristics of the new piezoelectric fan module were introduced in detail. The simulation results were compared by Flotherm software based piezoelectric fan module in off and on states.

Figure S1. RNA-sequencing for Notch1 over-expressing cells. A, The electrophoresis results of RNA extracted from the SACC cell samples implied a satisfactory RNA quality.1 N1 was the cells sample with NOTCH1 overexpressed vector while 1PS was the cells sample with negative vector, 3 N1 and 3PS were the duplicate samples of 1 N1 and 1PS. B, The reads of RNA-seq showed highly consistent in duplicate samples. C, Volcano plot for RNA-sequencing. Differentially expressed genes in each conditions were identified (in red or green), which is statistically significant. D, Heatmap for differentially expressed genes. Figure S2. The effect of HES1 knockdown on cell cycle. A, Representative images of Flow Cytometry analysis of cell cycle after PI staining. B, The percentage of cells in each phase of the cell cycle. Data from 2 independent experiments. (PDF 213 kb)

Objectives: Notch1 regulates tumor biology in a complex, context-dependent manner. The roles of Notch1 in tongue cancer are still controversial. The aim of this study is to investigate the roles of Notch1 in tongue cancer. Materials and Methods: The expression of Notch1 was tested between tongue cancer and normal samples by using immunohistochemistry. Tongue cancer cells were transfected with siRNA or plasmid, respectively. Cell proliferation, apoptosis, migration and invasion ability were tested in appropriate ways. The subcutaneous tumor model was established to observe the tumor growth. Results: Notch1 was upregulated in tongue carcinoma tissues and the expression of Notch1 was related with tumor stage and differentiation. Overexpression of Notch1 could increase tongue cancer cells proliferation, invasion and migration. But inhibited the expression of Notch1 could decrease cells proliferation, invasion and migration and promote cell apoptosis in vitro and in vivo. Conclusion: Our results prove that the oncogenic role of Notch1 in tongue cancer and provide the direction of targeted therapy of tongue cancer.

Several studies have reported that FZD2 regulates tumor biology in a complex manner. The aim of the present study was to identify the role of FZD2 in the cell growth and metastasis of salivary adenoid cystic carcinomas (SACCs). The expression of FZD2 in ACC-83 and ACC-LM cells were measured with real-time PCR. Immunohistochemical staining was used to detect the expression of FZD2 in clinical SACC samples with or without metastasis. Cell proliferation and Transwell assays were performed to explore the effects of FZD2 on cell growth and migration following the silencing of FZD2 with small interference RNAs and the overexpression of FZD2 with plasmid. Our data showed that FZD2 was downregulated in ACC-LM cells, which are an adenoid cystic carcinoma cell line with high metastatic potential, compared to ACC-83 cells, which have low metastatic potential. Additionally, the expression of FZD2 was lower in SACC tissues with metastasis compared to SACC tissues without metastasis (P

Background Our previous study demonstrated a close relationship between NOTCH signaling pathway and salivary adenoid cystic carcinoma (SACC). HES1 is a well-known target gene of NOTCH signaling pathway. The purpose of the present study was to further explore the molecular mechanism of HES1 in SACC. Methods Comparative transcriptome analyses by RNA-Sequencing (RNA-Seq) were employed to reveal NOTCH1 downstream gene in SACC cells. Immunohistochemical staining was used to detect the expression of HES1 in clinical samples. After HES1-siRNA transfected into SACC LM cells, the cell proliferation and cell apoptosis were tested by suitable methods; animal model was established to detect the change of growth ability of tumor. Transwell and wound healing assays were used to evaluate cell metastasis and invasion. Results We found that HES1 was strongly linked to NOTCH signaling pathway in SACC cells. The immunohistochemical results implied the high expression of HES1 in cancerous tissues. The growth of SACC LM cells transfected with HES1-siRNAs was significantly suppressed in vitro and tumorigenicity in vivo by inducing cell apoptosis. After HES1 expression was silenced, the SACC LM cell metastasis and invasion ability was suppressed. Conclusions The results of this study demonstrate that HES1 is a specific downstream gene of NOTCH1 and that it contributes to SACC proliferation, apoptosis and metastasis. Our findings serve as evidence indicating that HES1 may be useful as a clinical target in the treatment of SACC. Electronic supplementary material The online version of this article (10.1186/s12885-018-4350-5) contains supplementary material, which is available to authorized users.

// Bo-Hua Su 1,* , Jing Qu 1,* , Min Song 1,* , Xiao-Yu Huang 1 , Xiao-Meng Hu 1 , Jian Xie 1 , Yong Zhao 2 , Lin-Can Ding 1 , Lin She 1 , Jiang Chen 3 , Li-Song Lin 4 , Xu Lin 5 , Da-Li Zheng 5,6 and You-Guang Lu 1 1 Department of Preventive Dentistry, Affiliated Stomatological Hospital, Fujian Medical University, Fuzhou, China 2 Department of Pathology, Affiliated Stomatological Hospital, Fujian Medical University, Fuzhou, China 3 Center of Dental Implant, Affiliated Stomatological Hospital, Fujian Medical University, Fuzhou, China 4 Department of Oral and Maxillofacial Surgery, Affiliated First Hospital of Fujian Medical University, Fuzhou, China 5 Key Laboratory of Ministry of Education for Gastrointestinal Cancer, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, China 6 Molecular Oncology, Moffitt Cancer Center, Tampa, FL, United States * These authors contributed equally to this work Correspondence: Da-Li Zheng, email: // You-Guang Lu, email: // Keywords : NOTCH1, salivary adenoid cystic carcinoma, proliferation, apoptosis, metastasis Received : July 07, 2014 Accepted : August 05, 2014 Published : August 06, 2014 Abstract Background: Numerous studies have reported both the tumor-suppressive and oncogenic roles of the Notch pathway, indicating that Notch activity regulates tumor biology in a complex, context-dependent manner. The aim of the present study was to identify the role of NOTCH1 in the cell growth and metastasis of SACC. Methods: We analyzed the expression of NOTCH1 in clinical SACC samples using immunohistochemical staining. We silenced the expression of NOTCH1 and overexpressed activated NOTCH1 to elucidate the effects of NOTCH1 on proliferation, migration and invasion. NOTCH1 target genes were validated by real-time PCR. Results: Our results showed that NOTCH1 was upregulated in SACC tissues when compared with normal tissues, and this upregulation was further enhanced in SACC tissues with metastasis and recurrence when compared with SACC tissues without metastasis. Overexpression of NOTCH1 in SACC cells promoted cell growth, migration and invasion, and knockdown of NOTCH1 inhibited cell proliferation in vitro and tumorigenicity in vivo by inducing cell apoptosis. Conclusions:The results of this study suggest that NOTCH1 plays a key role in the cell growth, anti-apoptosis, and metastasis of SACC. NOTCH1 inhibitors might therefore have potential therapeutic applications in treating SACC patients by inhibiting cancer cell growth and metastasis.

Aberrant microRNA expression is involved in the regulation of various cellular processes, such as proliferation and metastasis in multiple diseases including cancers. MicroRNA‐30e‐5p (miR‐30e) was previously reported as an oncogenic or tumour suppressing miRNA in some malignancies, but its function in lung adenocarcinoma (LAC) remains largely undefined. In this study, we found that the expression of miR‐30e was increased in LAC tissues and cell lines, associated with tumour size and represented an independent prognostic factor for overall survival and recurrence of LAC patients. Further functional experiments showed that knockdown of miR‐30e suppressed cell growth while its overexpression promoted growth of LAC cells and xenografts in vitro and in vivo. Mechanistically, PTPN13 was identified as the direct target of miR‐30e in LAC, in which PTPN13 expression was down‐regulated in LAC tissues and showed the inverse correlation with miR‐30e expression. Overexpression of PTPN13 inhibited cell growth and rescued the proliferation‐promoting effect of miR‐30e through inhibition of the EGFR signalling. Altogether, our findings suggest that miR‐30e could function as an oncogene in LAC via targeting PTPN13 and act as a potential therapeutic target for treating LAC.

Dead-time effect results in distorted currents and force pulsations of Permanent Magnetic Linear Synchronous Motors(PMLSM),which limits the uses of precision stages in conditions of high accuracy,low frequency and lighter loadings.To compensate the Dead-time effect,this paper proposed a Double Updating Dead-time Compensation(DUDTC) method to correct each input pulse width of the motor driver according to phase current polarity.Firstly,the effect of dead-time on driver output pulses was analyzed and the correlation between correction of Pulse Width Modulation(PWM) and phase current polarities was given.Then,the method was evaluated through simulations in Matlab/Simulink and experiments in a linear-stage system.The results indicate that the phase error of DUDTC is only 0.5 servo periods and its phase current distortions is reduced.The DUDTC shows less codes and good portability,and is an efficient method to compensate the dead-time effect in digital drive systems.

Photodynamic therapy (PDT) has emerged as an alternative and promising noninvasive treatment for cancer as well as non-cancer diseases, which involves the uptake of photosensitizers (PSs) by cancer cells followed by irradiation. The use of nanomaterials as carriers of PSs is a very promising approach to improve the development of PDT in clinical medicine. In this study, a novel folic acid-conjugated graphene oxide (GO) was strategically designed and prepared as targeting drug delivery system to achieve higher specificity. The second generation photosensitizer (PS) Chlorin e6 (Ce6) was effectively loaded into the system via hydrophobic interactions and π-π stacking. The nanocarriers can significantly increase the accumulation of Ce6 in tumor cells and lead to a remarkable photodynamic efficacy on MGC803 cells upon irradiation. These suggested that folic acid-conjugated GO loaded Ce6 had great potential as effective drug delivery system in targeting PDT.

A vision based query interface annotation method is used to relate attributes and form elements in form-based web query interfaces, this method can reach accuracy of 82%. And a user participation method is used to tune the result; user can answer “yes” or “no” for existing annotations, or manually annotate form elements. Mass feedback is added to the annotation algorithm to produce more accurate result. By this approach, query interface annotation can reach a perfect accuracy.

Cadherins are found in almost all living organisms. In addition to their role in the formation and maintenance of normal tissue architecture, cadherins seem to play a crucial role in the cell-cell interactions of cancer cells in tumorigenesis, invasion and metastasis. The aim of the present study was to identify the role of CDH12 in the invasion and metastasis of salivary adenoid cystic carcinoma (SACC). Real-time PCR results showed that CDH12 is abnormally expressed in the highly metastatic SACC cell line ACC-M, compared to ACC-2, a SACC cell line with low metastatic ability. CDH12 expression was significantly higher in clinical samples with metastasis and recurrence than in those without metastasis and recurrence (P

The Notch signaling pathway is important for cell-cell communication; it is involved in gene regulation mechanisms that control multiple cell differentiation processes during embryonic and adult life. Notch is present in all metazoans, and vertebrates possess four different Notch receptors: Notch-1, Notch-2, Notch-3, and Notch-4. The aim of the present study was to identify the role of Notch protein in the metastasis of salivary adenoid cystic carcinoma (SACC). Real-time PCR results showed that Notch-1, Notch-2, and Notch-4 were upregulated in the highly metastatic SACC cell line ACC-M, compared to ACC-2, a SACC cell line with low metastatic ability. Knockdown of Notch-4 by small interfering RNA efficiently inhibited the invasion of ACC-M cells. Notch-4 expression was significantly higher in the clinical samples with metastasis and recurrence compared to that in control (p

To study the differential expression pattern of MMPs and TIMPs in ACC-M and ACC-2 tumor model.High and low metastatic tumor models were set up by transplantion of ACC-M and ACC-2 to nude mouse respectively. Then 3 mice in each group were selected randomly to detect the mRNA level of MMP-2, MMP-7, MMP-9, TIMP-1 and TIMP-2 using semiquantitative RT-PCR,GAPDH was used as internal control. Meanwhile, the expression of MMP-2, MMP-7, MMP-9, TIMP-1 and TIMP-2 in each transplanted tumor were determined by immunohistochemical assay, and IOD value of immunohistochemical reaction was measured. All data were analyzed by SPSS11.5 software package for Student's t test.The results of RT-PCR and immunohistochemistry showed an up-regulation of MMP-2, MMP-7, MMP-9 and TIMP-1 in ACC-M tumor model compared with ACC-2 model (P0.05), while the expression of TIMP-2 showed no significant difference between the two models (P0.05).The differential expression pattern of MMP-2, MMP-7, MMP-9, and TIMP-1 in 2 tumor models may result in different metastasis potential; The 2 tumor models provide good study model for investigation of the metastatic mechanism of salivary adenoid cystic carcinoma.

To construct differential expression profiles of adenoid cystic carcinoma cell lines for screening candidate genes related to metastasis and to verify some candidate genes in adenoid cystic carcinoma.Restriction fragments differential display PCR (RFDD-PCR) was used to set up gene expression profiles of adenoid cystic carcinoma cell lines-ACC-M and ACC-2, with high and low metastasis potential respectively. Candidate genes were screened through bioinformatics analysis. Then, a gene family of these candidate genes was checked using semi-quantitative reverse transcription-PCR(RT-PCR).Two gene expression profiles including 5420 gene fragments were constructed, 12 genes of a family called matrix metalloproteinase genes (MMPs) were observed obvious differentially expressed between two cell lines. Results of semi-quantitative RT-PCR also identified this different expression of MMP2,MMP7,MMP9,MMP14,MMP15 and MMP24.The construction of gene expression profiles of ACC-M and ACC-2 cell lines makes the foundation for seeking the target genes of adenoid cystic carcinoma. MMP2,MMP7,MMP9 and MMP15 may be relevant with carcinogenesis, development and metastasis of adenoid cystic carcinoma, and different metastasis potential may result from different subtype of MMPs gene family.