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Ardisia crispa(Myrsinaceae) is an ethnomedicine with horticultural and important medicinal values. Its morphology is complex, and its identification is difficult. We analyse the chloroplast genome characteristics and phylogenetic position of A. crispa to provide basic research data for the identification of A. crispa species and resource conservation. This study assemble and annotate the chloroplast genome of A. crispa and to compare it with the chloroplast genome within Ardisia. The A. crispa chloroplast genome is 156,785 bp in length, with a typical quadripartite structure containing 131 genes, including 86 protein-coding genes, 37 tRNA genes, and 8 rRNA genes; a total of 59 SSRs sites were identified, and the codon preference of this chloroplast genome is greater in A/U than in G/C, and leucine is the amino acid with the highest frequency of use. The chloroplast genomes of the nine Ardisia species are conserved in gene content and number, with more stable boundaries and less variation. In the phylogenetic tree, A. crispa is clustered on a branch with A. crispa var dielsii, and is closely related to A. mamillata and A. pedalis. In this study, we constructed and analyzed the chloroplast genome structure of A. crispa, and conducted phylogenetic analysis using the whole chloroplast genome sequence data of Ardisia plants, which is of great significance in understanding the genetic basis of A. crispa and adaptive evolution in Ardisia plants, and this will lay the foundation for the future research on A. crispa resource conservation and species identification., (© 2024. The Author(s).)

Primula secundiflora is an insect-pollinated, perennial herb belonging to the section Proliferae (Primulaceae) that exhibits considerable variation in its mating system, with predominantly outcrossing populations comprising long-styled and short-styled floral morphs and selfing populations comprising only homostyles. To facilitate future investigations of the population genetics and mating patterns of this species, we developed 25 microsatellite markers from P. secundiflora using next-generation sequencing and measured polymorphism and genetic diversity in a sample of 30 individuals from three natural populations. The markers displayed high polymorphism, with the number of observed alleles per locus ranging from three to 16 (mean = 8.36). The observed and expected heterozygosities ranged from 0.100 to 1.000 and 0.145 to 0.843, respectively. Twenty-one of the loci were also successfully amplified in P. denticulata. These microsatellite markers should provide powerful tools for investigating patterns of population genetic diversity and the evolutionary relationships between distyly and homostyly in P. secundiflora.

In this study, we describe and illustrate a new species, Primulaweiliei L.S.Yang, Z.K.Wu & G.W.Hu, from the Shennongjia Forestry District, Hubei Province in Central China. It is morphologically assigned to Primulasect.Aleuritia based on its dwarf and hairless habit, long petiole, fruits longer than calyx and covered by farina on the scape. This new species is similar to P.gemmifera and P.munroisubsp.yargongensis in the same section, but it can be distinguished by its smaller calyxes, homostylous flowers, corolla tube throat without annular appendage and only 1-2 flowers in each inflorescence. Based on the assessment conducted according to the IUCN Red List criteria, we propose that P.weiliei be classified as a Critically Endangered (CR) species., Competing Interests: The authors have declared that no competing interests exist., (Lin-Sen Yang, Zhi-Kun Wu, Hong-Wei Liang, Meng-Hua Zhang, Xian-Chun Zhang, Shuai Peng, Guang-Wan Hu.)

Ethyl acetate fraction of Toddalia asiatica was fractionated to yield fifteen previously undescribed prenylated coumarins, asiaticasics A-O (1-15) along with nine (16-24) known derivatives. The structures of these undescribed coumarins were established by spectroscopic analysis and reference data. Biological activity evaluation showed that compound 3 with the IC 50 value of 2.830 μM and compound 12 with the IC 50 value of 0.682 μM owned anti-inflammatory activity by detecting the rate of lactate dehydrogenase release in pyroptosis J774A.1 cells. The results showed that the expression of Caspase-1 and IL-1β was decreased in a dose-dependent manner in the compound 12 treatment group, suggesting that compound 12 may reduce pyroptosis by inhibiting NLRP3 inflammasome. To further determine that compound 12 treatment can inhibit macrophage pyroptosis, morphological observation was performed and the results were consistent with the bioactivity evaluation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Aim: To investigate the protective effects, antioxidant potential, and anti-inflammatory mechanisms of eicosane on glutamate-induced cell damage and on N-methyl-D-aspartate (NMDA)-induced retinal ganglion cell (RGC) injury in a mouse model of glaucoma., Methods: The protective effects of eicosane on the rat R28 retinal precursor cell line were assessed using cell counting kit-8 assays and Hoechst-propidium iodide staining. Intracellular reactive oxygen species (ROS) production was measured using the fluorescent probe 2'-7'-dichlorofluorescin diacetate and flow cytometry. The protective role of eicosane on NMDA-induced RGC injury in a mouse glaucoma model was determined by immunostaining of frozen sections of retina. The effects of eicosane on the metabolome of the retina in mice with NMDA-induced RGC damage were evaluated by liquid chromatography-mass spectroscopy (LC-MS) and untargeted metabolomics analyses., Results: Eicosane treatment significantly attenuated glutamate-induced damage to R28 cells in vitro . Eicosane also protected RGCs against NMDA-induced injury in a mouse glaucoma model. Untargeted metabolomics analyses showed that eicosane increased multiple metabolites, including L-arginine and L-carnitine, in the retina., Conclusion: Eicosane has protective effects, antioxidant potential, and anti-inflammatory properties in an in vitro model of glutamate-induced cell damage and in an in vivo model of NMDA-induced RGC injury in mouse glaucoma through modulation of L-arginine and/or L-carnitine metabolism., Competing Interests: Conflicts of Interest: Wu ZK, None; Li HY, None; Zhu YL, None; Xiong MQ, None; Zhong JX, None., (International Journal of Ophthalmology Press.)

Evolutionary transitions from outcrossing to selfing in flowering plants have convergent morphological and genomic signatures and can involve parallel evolution within related lineages. Adaptive evolution of morphological traits is often assumed to evolve faster than nonadaptive features of the genomic selfing syndrome. We investigated phenotypic and genomic changes associated with transitions from distyly to homostyly in the Primula oreodoxa complex. We determined whether the transition to selfing occurred more than once and investigated stages in the evolution of morphological and genomic selfing syndromes using 22 floral traits and both nuclear and plastid genomic data from 25 populations. Two independent transitions were detected representing an earlier and a more recently derived selfing lineage. The older lineage exhibited classic features of the morphological and genomic selfing syndrome. Although features of both selfing syndromes were less developed in the younger selfing lineage, they exhibited parallel development with the older selfing lineage. This finding contrasts with the prediction that some genomic changes should lag behind adaptive changes to morphological traits. Our findings highlight the value of comparative studies on the timing and extent of transitions from outcrossing to selfing between related lineages for investigating the tempo of morphological and molecular evolution., (© 2024 The Authors New Phytologist © 2024 New Phytologist Foundation.)

Background: This study was designed to compare the body fluid module of Sysmex XN9000 (XN-BF) with optical microscopy (OM) for cerebrospinal fluid (CSF) analysis after two-step cell slide centrifuge (TSCSC), defining the best procedure for CSF optical microscopy analysis., Methods: Items of RBC, WBC enumeration and differentiation were observed. The cell count and morphologic evaluation of the cellular composition by OM was carried out both with and without two-step cell slide centrifuge (TSCSC) and were compared the data with XN-BF., Results: There were 69.98 ± 4.94 RBC and 36.98 ± 3.39 WBC in one OSCSC microscopic field whereas there were 96.35 ± 5.41 RBC and 66.15 ± 4.85 WBC in one TSCSC microscopic field in the same sample (*200). There was a statistical difference between those two methods (p = 0.000). Excellent correlation was found between total cell count with both OM and XN-BF. The R2 value for RBC and WBC counts were 0.99 and 0.96, respectively. For WBC differential, the R2 values were 0.98 for PMN and 0.70 for MN. Correlation of MN was poorer than PMN. As far as the tumor cell, phagocyte, and plasma cell with high fluorescence were concerned, OM were not consistent with XN-BF., Conclusions: The TSCSC procedure contributes to the separation of cells and other ingredients. XN-BF displays excellent performance at RBC and WBC cell count except for mononuclear cells, tumor cells, phagocytes, and leukemia cells. which makes it just a practical alternative to total cell (WBC, RBC) count for CSF samples. Detailed morphologic workup of CSF samples is mandated in all cases with meningoencephalitis, elevated cell count, sub-arachnoid hemorrhage and meningeal carcinomatosis, the TSCSC procedure is recommended.

Objective: To compare the clinical efficacy between closed reduction combined with semi-circular external fixator and minimally invasive percutaneous plate osteosynthesis (MIPPO) in the treatment of middle anddistal tibia fractures., Methods: The clinical data of sixty patients with middle and distal tibia fractures admitted between January 2019 and November 2022, were retrospectively analyzed. These patients were categorized into external fixation group ( n =30) and internal fixation group ( n =30). There were 18 males and 12 females in the external fixation group, with an average age of (49.29±2.35) years old. Among them, 14 patients presented with fractures on the left side, and 16 patients presented with fractures on the right side. Closed reduction, arched wire, and semi-circular external fixator were used for treatment. There were 20 males and 10 females in the internal fixation group, with an average age of (48.96±1.87) years old. Among them, 15 patients presented with fractures on the left side, and 15 patients presented with fractures on the right side. MIPPO technique was used for the treatment. Perioperative parameters, including time injury to surgery, surgical duration, incision length, intraoperative bleeding, time to active activity, and incision healing level, were compared between the two groups. Clinical outcomes were also assessed, including Johner-Wruhs scores, time to minimum pain-adapted full weight-bearing, visual analog scale (VAS), SF-36 scale, and complications., Results: The external fixation group exhibited a significantly shorter incision length (1.36±0.86) cm and lower intraoperative bleeding (10.83±5.73) ml compared to the internal fixation group (12.74±3.12) cm and (86.47±8.90) ml, respectively( P <0.05). The postoperative active activity time (1.50±0.54) days and minimum pain-adapted full weight-bearing activity time(108.87±3.43) days in the external fixation group were slightly delayed than the internal fixation group(1.15±0.98) days and (105.27±3.68) days, respectively( P <0.05). Over a mean postoperative follow-up duration of (6.23±1.89) months, both groups showed improved VAS and SF-36 scale scores. There were no statistically significant differences in VAS and SF-36 scale scores 1, 3, 6 months post-operatively between the two groups( P >0.05). The intraoperative surgical time in the external fixation group (35.42±9.31) minutes was shorter than that in the internal fixation group(74.22±7.81) minutes ( P <0.05). There was no intraoperative vascular or nerve injury, nor postoperative skin necrosis in the external fixation group. However, skin necrosis was observed in 6 patientsin the internal fixation group, representing a statistically significant difference ( P <0.05)., Conclusion: Both external fixation and plate internal fixation are effective methods for the treatment of middle and distal tibia fractures. External fixation exhibits the advantage of less surgical trauma and a lower incidence of complications.

Four new diterpenoids (1-4) and sixteen known diterpenoids (5-20) were purified from the whole plant of Euphorbia helioscopia L. Compounds 1 and 2 were rhamofolane diterpenoids with a 5/7/6 tricyclic systems, compound 3 was a lathyranes diterpenoid, and compound 4 was a jathophanes diterpenoid. The isolated compounds were tested for their cytotoxicity and anti-Zika virus properties, and compounds 9 and 15 showed low cytotoxicity and strong anti-Zika virus properties with EC 50 2.63 and 5.94 μM, respectively. Further, the inhibitory effects of compounds on protein levels were determined using Western blotting and immunofluorescence assays., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2023. Published by Elsevier B.V.)

Histone modification plays an important role in pathological cardiac hypertrophy and heart failure. In this study we investigated the role of a histone arginine demethylase, Jumonji C domain-containing protein 6 (JMJD6) in pathological cardiac hypertrophy. Cardiac hypertrophy was induced in rats by subcutaneous injection of isoproterenol (ISO, 1.2 mg·kg -1 ·d -1 ) for a week. At the end of the experiment, the rats underwent echocardiography, followed by euthanasia and heart collection. We found that JMJD6 levels were compensatorily increased in ISO-induced hypertrophic cardiac tissues, but reduced in patients with heart failure with reduced ejection fraction (HFrEF). Furthermore, we demonstrated that JMJD6 overexpression significantly attenuated ISO-induced hypertrophy in neonatal rat cardiomyocytes (NRCMs) evidenced by the decreased cardiomyocyte surface area and hypertrophic genes expression. Cardiac-specific JMJD6 overexpression in rats protected the hearts against ISO-induced cardiac hypertrophy and fibrosis, and rescued cardiac function. Conversely, depletion of JMJD6 by single-guide RNA (sgRNA) exacerbated ISO-induced hypertrophic responses in NRCMs. We revealed that JMJD6 interacted with NF-κB p65 in cytoplasm and reduced nuclear levels of p65 under hypertrophic stimulation in vivo and in vitro. Mechanistically, JMJD6 bound to p65 and demethylated p65 at the R149 residue to inhibit the nuclear translocation of p65, thus inactivating NF-κB signaling and protecting against pathological cardiac hypertrophy. In addition, we found that JMJD6 demethylated histone H3R8, which might be a new histone substrate of JMJD6. These results suggest that JMJD6 may be a potential target for therapeutic interventions in cardiac hypertrophy and heart failure., (© 2023. The Author(s), under exclusive licence to Shanghai Institute of Materia Medica, Chinese Academy of Sciences and Chinese Pharmacological Society.)

Primulajiaozishanensis Z.K.Wu, W.H.Yang & Yuan Wu, a new species of Primulaceae is described and illustrated from Jiaozi Snow Mountain of Dongchuan District, Yunnan, China. Morphological evidence supports P.jiaozishanensis as a member of P.sect.Petiolaressubsect.Davidii, which is characterized by firmly papery or leathery leaves, with veins impressed adaxially, often prominently raised and alveolate abaxially. The new species is characterized by having long and stout rhizomes, smaller leaves with short petioles, short or almost obsolete scape, and larger flowers. The distribution, phenology and conservation status of the new species are also provided., Competing Interests: No conflict of interest was declared., (Yuan Wu, Wu-Hai Yang, Zhi-Kun Wu.)

The rare Primulabrachystoma W.W.Sm. is an endemic species confined to Gaoligong mountain of Chinese-Burma frontier, which has been rediscovered from the same region after nearly 100 years. In total, 11 specimens from Gaoligong Mountain have been found in the herbaria worldwide, since its first collection in 1920 by Farrer, Reginald John. Previously, this species was described as homostylous but our finding shows the species also exhibited heterostyly. A complete description of the species, the distribution, morphological comparison and identification key from closely related species are provided here. An assessment of its conservation status suggests that the species is 'Endangered' (EN)., Competing Interests: No conflict of interest was declared., (Xiao-Qi Jiang, Na Zhang, Zhi-Kun Wu.)

Three new rearranged diterpenoids, strophioblachins A-C ( 1 - 3 ), eight new diterpenoids, strophioblachins D-K ( 4 - 11 ), and seven previously described diterpenoids ( 12 - 18 ) were purified from the aerial parts of Strophioblachia fimbricalyx. Compounds 1 and 2 contain a rare 6/6/5/6 ring system, while 3 has an uncommon tricyclo[4.4.0.0 8,9 ]tridecane-bridged unit, and their diterpenoid skeletons are being reported for the first time. Utilizing spectroscopic and HRESIMS data analysis, the structures of the new compounds ( 1 - 11 ) were established, and ECD and 13 C NMR calculations were used to confirm the relative and absolute configurations of 11 and 9 . The absolute configurations of compounds 1 , 3 , and 10 were established using single-crystal X-ray diffraction. The results of testing for anticardiac hypertrophic activity demonstrated that compounds 10 and 15 dose-dependently lowered the mRNA expression of Nppa and Nppb . Protein levels were confirmed by Western blotting, which also demonstrated that compounds 10 and 15 lowered the expression of the hypertrophic marker ANP. The cytotoxic activity against neonatal rat cardiomyocytes was assayed in vitro by the CCK-8 and ELISA methods, and the results showed that compounds 10 and 15 were only very weakly active in the range.

Primulapingbaensis Na Zhang, X.Q.Jiang & Z.K.Wu, a new species of Primulaceae from Gaofeng Mountain of Pingba county, Guizhou, China, is described and illustrated. Morphological evidence supports P.pingbaensis as a member of P.sect.Petiolares on account of scape elongating, pedicels conspicuously thickening in fruit, and its capsule cracking irregularly round the top and crumbling away. Amongst the members of subsect. Davidii, the new species is characterized by having a uniquely smooth leaf blade due to inconspicuously raised veinlets and homostylous flowers with the style usually extending beyond the anthers. The distribution, phenology and conservation status of the new species are also provided., (Na Zhang, Xiao-Qi Jiang, Zhi-Kun Wu.)

Purposes: Temporary hemiepiphysiodesis (TH) using eight-plates is one of the most frequently performed surgeries for correcting angular deformities of the lower extremities in adolescents. Rarely have studies examined children with X-linked hypophosphataemic rickets (X-LHPR) treated with TH using eight-plates. This study was conducted to investigate the efficacy, the endpoint, and the complications of TH using eight-plates to correct angular deformities of the lower extremities in skeletally immature children., Methods: We reviewed a total of 26 children (86 physes, 52 knees) with X-LHPR (mean age of 6.2 years, range from 2 to 13 years) who underwent TH using eight-plate to correct angular deformities of the lower extremities. Radiographs and clinical records of these patients were evaluated for demographic data and related clinical factors., Results: The average correction of the mechanical lateral distal femoral angle (mLDFA) was 11.7 ± 8.7° (range from 1.0 to 29.7°), and the average correction of the mechanical medial proximal tibial angle (mMPTA) was 8.4 ± 5.0° (range from 0.3 to 16.7°). The mean deformity correction time was 22.7 months (range from 7 to 60 months), and the mean follow-up after eight-plate removal was 43.9 months (range from 24 to 101 months). Overall, 76.9% (20/26 patients) of the angular deformities of the knee were completely corrected and 15.4% (4/26) of the patients received osteotomy surgery. The femoral correction velocity (0.9° per month) was significantly higher than the proximal tibial (0.6° per month) (p = 0.02). The correction velocity of the mLDFA and mMPTA with the TH procedure was faster than that in the absence of intervention (0.9° vs. 0.2°, 0.7° vs. 0.4° per month, p < 0.05). The correction velocity of the mLDFA (1.2° vs. 0.5° per month, [Formula: see text]) and mMPTA (0.7° vs. 0.5° per month, p = 0.04) of patients whose age ≤ five years old was faster than that of patients whose age > five years old. A total of 69.2% (18/26) patients experienced one TH procedure using eight-plates only. Two patients had screw loosening (2/26, 7.7%). One patient (1/26, 3.8%) had a rebound phenomenon after the removal of eight-plate and had the TH procedure again. There was no breakage, infection, physis preclosure, or limited range of movement found in the follow-up., Conclusion: TH using eight-plates is a safe and effective procedure with a relatively low incidence of complication and rebound, and it could be used as part of a streamlined treatment for younger X-LHPR patients with resistant or progressive lower limb deformity despite optimal medical treatment. Early intervention can achieve better results., (© 2023. The Author(s) under exclusive licence to SICOT aisbl.)

A new species, Primula luquanensis Z.K.Wu and Wei Zhou sp. nov. (Primulaceae) is described and illustrated from Yunnan Province, China. It is morphologically assigned to P. sect Aleuritia based on its dwarf and hairless habit and coverage by farina on both sides of the leaf blade and scape. This new species is similar to P. nutantiflora and P. yunnanensis , but it is easily distinguished by its stolons, solitary bract, bell-shaped corolla and monomorphic floral form. The new species also has a substantially reduced corolla tube, presenting a unique floral form in a genus where heterostyly typically prevails.

Background: The persistence of symptoms in patients with chronic neck pain is considered to be associated with variation in the neck muscle structure and associated neuromuscular control. Sling exercise therapy (SET) has been demonstrated to relieve the symptoms of chronic neck pain, whereas it is controversial whether this benefit is correlated to altered neck muscle structure and associated neuromuscular control in the patients., Objective: To investigate the effect of SET on cervical muscle structure (thickness) and associated neuromuscular control in patients with chronic neck pain., Methods: Twenty-five patients with chronic neck pain were randomly assigned to the SET group (n= 12) or the control group (n= 13). The SET group received the SET intervention for 4 weeks, while the control group maintained normal activities of daily living. At baseline and after 4 weeks of intervention, Visual analogue scale and neck disability index were measured in both groups, and changes in the thickness of the superficial cervical muscles were assessed using musculoskeletal ultrasound. Surface electromyography (EMG) was adapted to assess the neuromuscular control of the neck while the participant was performing the cranio-cervical flexion test., Results: At 4 weeks, the SET group had a significant reduction of RMS in both UT and SCM of EMG compared to the control group (p< 0.05). Regarding ultrasound, the SET group had significantly lower muscle thickness compared to the control group in both the rest position and the MVIC position (p< 0.05). There were no within-group differences in the control group (p> 0.05), while the SET group showed significant reductions in both RMS and muscle thickness (p< 0.05)., Conclusion: 4-week SET was effective in reducing pain and dysfunction in patients with chronic neck pain, which may be related to improved neck muscle thickness and neuromuscular control of the neck.

Vaccination remains the most effective and cost-saving measure to protect against hepatitis B, a global health problem. It is crucial to characterize the persistence of the immune response after booster vaccination. This study aimed to quantify the persistence through mathematical modeling. Booster vaccination against hepatitis B was conducted in children 5-15 years in 2009-10 in Zhejiang Province. There were four dosage formulations of hepatitis B vaccines [Shenzhenkangtai Biotechnology Co. Ltd. Dalianhanxin Biotechnology Co. Ltd. NCPC GeneTech Biotechnology Pharmaceutical Co. Ltd. Sinovac Biotech Co. LTD. China]: 5, 10, and 20 μg hepatitis B vaccines or 5 μg hepatitis A and B (HAB) combination vaccine with a 0-1-6-month schedule. These were randomly administered to children negative for all hepatitis B markers, named as the schedule 2 group. Anti-HBs positive subjects were given one dose of booster, named as the schedule 1 group. Anti-HBs antibody was measured 1, 7, 18, 66, and 102 months after the first booster dose. A linear mixed-effects model was proposed to predict long-term persistence. One hundred two months after the booster dose, the mean anti-HBs levels were 33.8 mIU/mL, with 73.7 mIU/mL for the schedule 1 group and 20.2 mIU/mL for the schedule 2 group. The model predicted that 99.5% of subjects would remain seropositive (≥10mIU/mL) at year 20 post booster vaccination, with 100.0% and 98.8% for the schedule 1 group and the schedule 2 group, respectively, whereas at year 30, the seropositivity rates would decrease to 76.8%, with 99.4% for the schedule 1 group and 62.5% for the schedule 2 group. The immunogenicity of the booster vaccination could persist for at least 8 years. Mathematical modeling may predict even longer, up to 30 years of protection.

Cardiac fibrosis is thought to be the hallmark of pathological hypertrophic remodeling, of which the myofibroblast transdifferentiation is the key cell biological event. However, there is still no specific and effective therapeutic agent approved for cardiac fibrosis. To investigate the effects of belumosudil, the first ρ-associated kinase-2 (ROCK2)-specific inhibitor, on cardiac hypertrophy, fibrosis, and dysfunction induced by pressure overload, the transverse aortic constriction (TAC) or sham operation was carried out on wild-type C57BL/6 mice (male, 6-8 wk old) under pentobarbital anesthesia. After that, mice were randomly divided into three groups: sham operation + vehicle, TAC + vehicle, TAC + 50 mg·kg -1 ·day -1 belumosudil. We found that belumosudil effectively ameliorated cardiac hypertrophy, fibrosis, and dysfunction in TAC mice. To elucidate the underlying mechanism, we inhibited the expression of ROCK2 in vitro by either belumosudil or siRNA. We showed that the inhibition of ROCK2 by either belumosudil or knockdown suppressed cardiac fibroblasts activation and proliferation significantly induced by transforming growth factor-β1 (TGF-β1). Furthermore, our study confirmed ROCK2 mediates cardiac fibrosis by interacting with TGF-β1/mothers against decapentaplegic homolog 2 (Smad2) pathway. Taken together, we demonstrated that belumosudil ameliorates cardiac hypertrophy and fibrosis induced by TAC via inhibiting cardiac fibroblasts activation. In conclusion, belumosudil may be a promising therapeutic drug for cardiac hypertrophy and fibrosis induced by myocardial pressure overload. NEW & NOTEWORTHY Although ρ-associated kinase-2 (ROCK2) is the main isoform of ρ-associated kinases (ROCKs) in the heart and more important in cardiac hypertrophy and fibrosis than ρ-associated kinase-1 (ROCK1), there has not been any pharmacological approach to inhibit ROCK2 selectively. Our study demonstrates for the first time that belumosudil, the first ROCK2-specific inhibitor, effectively ameliorates cardiac hypertrophy, fibrosis, and dysfunction induced by TAC via inhibiting cardiac fibroblasts activation.

Background The early mortality after surgery for infective endocarditis is high. Although risk models help identify patients at high risk, most current scoring systems are inaccurate or inconvenient. The objective of this study was to construct an accurate and easy-to-use prediction model to identify patients at high risk of early mortality after surgery for infective endocarditis. Methods and Results A total of 476 consecutive patients with infective endocarditis who underwent surgery at 2 centers were included. The development cohort consisted of 276 patients. Eight variables were selected from 89 potential predictors as input of the XGBoost model to train the prediction model, including platelet count, serum albumin, current heart failure, urine occult blood ≥(++), diastolic dysfunction, multiple valve involvement, tricuspid valve involvement, and vegetation >10 mm. The completed prediction model was tested in 2 separate cohorts for internal and external validation. The internal test cohort consisted of 125 patients independent of the development cohort, and the external test cohort consisted of 75 patients from another center. In the internal test cohort, the area under the curve was 0.813 (95% CI, 0.670-0.933) and in the external test cohort the area under the curve was 0.812 (95% CI, 0.606-0.956). The area under the curve was significantly higher than that of other ensemble learning models, logistic regression model, and European System for Cardiac Operative Risk Evaluation II (all, P <0.01). This model was used to develop an online, open-access calculator (http://42.240.140.58:1808/). Conclusions We constructed and validated an accurate and robust machine learning-based risk model to predict early mortality after surgery for infective endocarditis, which may help clinical decision-making and improve outcomes.

Jumonji domain-containing 3 (JMJD3/KDM6B) is a histone demethylase that plays an important role in regulating development, differentiation, immunity, and tumorigenesis. However, the mechanisms responsible for the epigenetic regulation of inflammation during mastitis remain incompletely understood. Here, we aimed to investigate the role of JMJD3 in the lipopolysaccharide (LPS)-induced mastitis model. GSK-J1, a small molecule inhibitor of JMJD3, was applied to treat LPS-induced mastitis in mice and in mouse mammary epithelial cells in vivo and in vitro. Breast tissues were then collected for histopathology and protein/gene expression examination, and mouse mammary epithelial cells were used to investigate the mechanism of regulation of the inflammatory response. We found that the JMJD3 gene and protein expression were upregulated in injured mammary glands during mastitis. Unexpectedly, we also found JMJD3 inhibition by GSK-J1 significantly alleviated the severity of inflammation in LPS-induced mastitis. These results are in agreement with the finding that GSK-J1 treatment led to the recruitment of histone 3 lysine 27 trimethylation (H3K27me3), an inhibitory chromatin mark, in vitro. Furthermore, mechanistic investigation suggested that GSK-J1 treatment directly interfered with the transcription of inflammatory-related genes by H3K27me3 modification of their promoters. Meanwhile, we also demonstrated that JMJD3 depletion or inhibition by GSK-J1 decreased the expression of toll-like receptor 4 and negated downstream NF-κB proinflammatory signaling and subsequently reduced LPS-stimulated upregulation of Tnfa, Il1b, and Il6. Together, we propose that targeting JMJD3 has therapeutic potential for the treatment of inflammatory diseases., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Primulalongipilosa from SW Yunnan, China, is described as a species new to science and illustrated. The systematic placement of this new species is also discussed based on an nrITS molecular tree. It is morphologically most similar to P.mollis , but differs from the latter in its racemose inflorescence, green calyx tube, pink to pink rose corolla, stamens at 1/3 length above the base of the corolla tube and applanate globose capsule., (Ze-Huan Wang, Yi Wang, Li Chen, Hua Peng, Zhi-Kun Wu, Guang Guo.)

Background: Evaluation of intra-articular osteochondral fractures in children with acute traumatic lateral patellar dislocation (LPD) is important for determining treatment options., Purpose: To (1) compare the diagnostic accuracy of computed tomography (CT) and magnetic resonance imaging (MRI) for evaluating intra-articular osteochondral fractures; (2) compare the interpretation of CT and MRI images between radiologists and pediatric orthopaedic surgeons (POS); and (3) investigate any clinical factors influencing the accuracy of CT and MRI evaluations., Study Design: Cohort study (diagnosis); Level of evidence, 2., Methods: We reviewed 35 knees in 35 patients (mean age, 12.2 ± 1.2 years; 12 boys and 23 girls) who were treated arthroscopically for acute traumatic LPD; 71% of the patients had patellar fractures, 54% had femoral fractures, and 60% had free osteochondral fracture fragments. All presurgical MRI and CT images were reviewed by POS who were blinded to both the reports of the radiologists and surgical records. We compared the accuracy of CT and MRI in diagnosing intra-articular osteochondral fractures against the arthroscopic findings and compared the interpretation of the images by the POS (MRI-O, CT-O) with those of the radiologists (MRI-R, CT-R)., Results: There was no significant difference in diagnostic accuracy between CT and MRI for overall intra-articular osteochondral fractures by the POS or the radiologists; however, the CT-O images had a higher diagnostic specificity (84.2% vs 69.6%; P < .001) and sensitivity (88.1% vs 70.1%; P < .001) versus the MRI-R images. Regarding free fracture fragments, the CT-R images had a higher diagnostic accuracy than the MRI-R images (73.5% vs 47.1%; P = .026). When backed by clinical data, the MRI-O images had greater diagnostic accuracy (78.7% vs 60.3%; P = .001) and sensitivity (88.1% vs 30.7%; P = .021) but lower specificity compared with the MRI-R images, and the CT-O images had similar diagnostic accuracy but greater sensitivity than the CT-R images (70.1% vs 52.2%; P < .001). The diagnostic accuracy of MRI-O images was lower for children under 12 years versus children 12 years and over (67.5% vs 83.3%; P = .040)., Conclusion: Compared with MRI, CT scans had better diagnostic performance in the evaluation of intra-articular osteochondral fractures in pediatric patients with acute traumatic LPD. Clinical data enhanced the diagnostic sensitivity of MRI and CT but decreased the specificity of MRI. MRI evaluations remain challenging for both POS and radiologists., Competing Interests: One or more of the authors has declared the following potential conflict of interest or source of funding: This work was supported by Shanghai Collaborative Innovation Center for Translational Medicine (grant TM201712) and Interdisciplinary of Medicine and Engineering of University of Shanghai for Science and Technology (grant 10-21-308-406). AOSSM checks author disclosures against the Open Payments Database (OPD). AOSSM has not conducted an independent investigation on the OPD and disclaims any liability or responsibility relating thereto., (© The Author(s) 2022.)

Indigofera stachyodes Lindl. is a traditional medicinal plant in southwestern China. In this study, we report the complete chloroplast genome sequence of I. stachyodes , using next-generation sequencing technology. The complete chloroplast genome of I. stachyodes was 158,039 bp in length with an overall GC content 35.80%, containing a large single-copy (LSC) region of 88,772 bp, a small single-copy (SSC) region of 18,733 bp, and a pair of inverted repeats (IRs) regions of 25,267 bp. In total, there are 128 genes (83 protein-coding genes (PCGs), eight ribosomal RNA (rRNA) genes, and 37 tRNA genes) in the whole chloroplast genome, including 113 unique genes (78 unique PCGs, 31 unique tRNAs, and four unique rRNAs). The phylogenetic analysis indicated that I. stachyodes formed a monophyletic clade with I. tinctoria and I. linifolia , showing that they have close relationship. The complete chloroplast genome of I. stachyodes provides valuable genomic information for the phylogeny, molecular identification and sustainable utilization of this species., Competing Interests: No potential conflict of interest was reported by the authors., (© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Periploca forrestii Schltr. is a traditional medicine plant in southwestern China. In this study, we characterize the complete chloroplast (cp) genome of P. forrestii based on next-generation sequencing. The cp genome is 154,140 bp in size with an overall GC content 38.2%, including a large single-copy (LSC) region (84,941 bp), a small single-copy (SSC) region of 17,619 bp, and two inverted repeats (IRs) regions, each of 25,790 bp. A total of 130 genes (85 protein-coding genes, 8 ribosomal RNA (rRNA) genes, and 37 transfer RNA (tRNA genes)) are annotated in the whole chloroplast genome, containing 113 unique genes (79 unique CDSs, 30 unique tRNAs, and 4 unique rRNAs). The phylogenetic analysis indicated that P. forrestii formed a monophyletic clade with the same genus plant P. sepium , showing that they have close relationship. The complete chloroplast genome of P. forrestii provides valuable genomic information for the phylogeny, molecular identification and sustainable utilization of this species., Competing Interests: No potential conflict of interest was reported by the authors., (© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Artemisinin, a sesquiterpene lactone widely used in malaria treatment, was discovered in the medicinal plant Artemisia annua. The biosynthesis of artemisinin is efficiently regulated by jasmonate (JA) and abscisic acid (ABA) via regulatory factors. However, the mechanisms linking JA and ABA signalling with artemisinin biosynthesis through an associated regulatory network of downstream transcription factors (TFs) remain enigmatic. Here we report AaTCP15, a JA and ABA dual-responsive teosinte branched1/cycloidea/proliferating (TCP) TF, which is essential for JA and ABA-induced artemisinin biosynthesis by directly binding to and activating the promoters of DBR2 and ALDH1, two genes encoding enzymes for artemisinin biosynthesis. Furthermore, AaORA, another positive regulator of artemisinin biosynthesis responds to JA and ABA, interacts with and enhances the transactivation activity of AaTCP15 and simultaneously activates AaTCP15 transcripts. Hence, they form an AaORA-AaTCP15 module to synergistically activate DBR2, a crucial gene for artemisinin biosynthesis. More importantly, AaTCP15 expression is activated by the multiple reported JA and ABA-responsive TFs that promote artemisinin biosynthesis. Among them, AaGSW1 acts at the nexus of JA and ABA signalling to activate the artemisinin biosynthetic pathway and directly binds to and activates the AaTCP15 promoter apart from the AaORA promoter, which further facilitates formation of the AaGSW1-AaTCP15/AaORA regulatory module to integrate JA and ABA-mediated artemisinin biosynthesis. Our results establish a multilayer regulatory network of the AaGSW1-AaTCP15/AaORA module to regulate artemisinin biosynthesis through JA and ABA signalling, and provide an interesting avenue for future research exploring the special transcriptional regulation module of TCP genes associated with specialized metabolites in plants., (© 2021 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)

The aberrant methylation of many genes has been reported to be associated with various carcinomas. Accurate detection of the methylation level could provide critical insights into the diagnostic analysis of diseases. Here, a sensitive HpaII-edited absolute droplet loop-mediated isothermal amplification (HEADLAMP) method based on methylation-sensitive restriction enzyme (MSRE) HpaII was developed for the digital quantification of DNA methylation. Methylation levels of the death-associated protein kinase 1 (DAPK1) gene that is associated with many cancers were studied using β-actin as an internal reference. DAPK1 (2.5 pM) with 0.01% methylation (250 aM) can be detected with the conventional HpaII-edited LAMP assay. Using HEADLAMP, as low as 1% methylation level can be distinguished with an estimated limit of detection of 5 aM (ca. 3 copies/μL). Moreover, HEADLAMP can detect low levels of methylated DAPK1 in normal L-02 cells, while the conventional assay cannot. Finally, HEADLAMP was applied to the detection of DAPK1 methylation in 20 clinical tissue samples, which revealed hypermethylated DAPK1 in cervical cancer patients. We envisage potential applications of this robust, specific, and sensitive HEADLAMP assay in epigenetic studies and early clinical diagnosis.

Background and purpose - Proximal femoral osteotomy (PFO) is commonly performed to treat children with developmental dysplasia of the hip (DDH). Implant-related femoral fractures after osteotomy are sometimes reported, but the potential risk factors for these fractures remain unclear. We investigated the association of implant-related fractures with PFO and potential risk factors for these fractures.Patients and methods - We retrospectively reviewed 1,385 children undergoing PFO for DDH in our institution from 2009 to 2016 after obtaining institutional review board (IRB) approval and identified 27 children (28 hips, fracture group) with implant-related femoral fractures after PFO. We selected 137 children (218 hips, control group) without fractures who matched the children in the fracture group by age, weight, surgeon, and surgical period. Relevant clinical data were collected and compared between the 2 groups. Multiple analyses of risk factors for implant-related fractures were conducted by logistic regression with the stepwise regression method.Results - The occurrence rate of implant-related fractures was 1.9% (27/1,385). Compared with the control group, the fracture group more commonly exhibited bilateral involvement (74% vs. 53%, p = 0.04), used a spica orthosis for immobilization after osteotomy (43% vs 21%, p = 0.01) and exhibited mild remodeling at the osteotomy site (46% vs. 19%, p = 0.003), and less commonly required capsulotomy during osteotomy (61% vs. 79%, p = 0.03). According to the multiple regression analysis, the only factor identified as an independent risk factor for implant-related fractures was mild remodeling at the osteotomy site (OR = 3.2, 95% CI 1.4-7.5). Remodeling at the osteotomy site was significantly associated with varus osteotomy (coefficient = 1.4, CI 1.03-1.8). The fracture occurred at a mean of 12 months (2.2-25) after osteotomy or 3.3 months (0-12) after implant removal. In children undergoing implant removal, the fractures mostly occurred at the osteotomy site (n = 13/15), while in those with the implant remaining, the fractures mostly occurred in the screw hole (n = 8/13).Interpretation - The type of PFO performed is not associated with implant-related fractures in children with DDH. Children with mild remodeling at the osteotomy site should be closely followed up, regardless of whether the hardware is removed, and high-intensity activity should not be permitted until moderate or extensive remodeling is confirmed. After PFO, the implants should be removed when solid union is achieved at the osteotomy site.

Primula homogama F. H. Chen & C. M. Hu (Primulaceae) is endemic to the Emei Mountain of China. In this study, we characterized the complete chloroplast genome of P. homogama based on next-generation sequencing (NGS). The complete chloroplast genome of P. homogama was 154,677 bp in size with a typical quadripartite structure, containing a large single-copy (LSC) region of 85,299 bp and a small single-copy (SSC) region of 17,816 bp. These two regions were separated by a pair of inverted repeat regions (IRs), each of 25,781 bp. A total of 130 functional genes were encoded, consisted of 86 protein-coding genes (PCG), 36 tRNA genes, and eight ribosomal RNA (rRNA) genes., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Background: Temporary hemiepiphysiodesis (TH) using an eight-Plate implant is one of the most common surgeries used for the correction of coronal angular deformities around the knee in adolescents. However, few studies have focused on children aged less than 10 years treated with TH using an eight-Plate implant. The purpose of this study was to investigate the efficacy, correction velocity, and complications of TH with an eight-Plate implant as well as the occurrence of rebound and risk factors in this population., Methods: This retrospective study included a total of 135 physes (101 knees) from 66 children (mean age of 4.69 years old, range from 1 to 10 years old) who underwent TH with an eight-Plate implant to correct coronal genu angular deformities in our hospital. Related clinical factors were recorded and analysed by multivariable linear and logistic regression models., Results: The mean deformity correction period was 13.26 months, and the mean follow-up after eight-Plate removal was 12.71 months. In all, 94.06% (95/101 knees) of the genu angular deformities were completely corrected. Non-idiopathic genu angular deformity was found to be an independent risk factor for deformity correction failure (odds ratio (OR) = 2.47). The femoral correction velocity was significantly higher than the tibial correction velocity (1.28° vs. 0.83° per month, p < 0.001). After adjustment for other factors, younger children had higher correction velocities in the distal femur; however, genu valgum and idiopathic deformities were associated with higher correction velocities in the proximal tibia. In addition, we found three (3/101, 2.97%) knees with genu valgum that experienced rebound after removal of the eight-Plate, while five (5/101, 4.95%) knees with non-idiopathic genu angular deformity experienced screw loosening. No other complications were found, and non-idiopathic deformity was the only risk factor for complications (OR = 3.96). No risk factor was found for rebound in our study., Conclusions: TH using an eight-Plate implant is an effective procedure for coronal genu angular deformities with a low incidence of complications and rebound in patients younger than 10 years old. For this population, TH using an eight-Plate should be considered as soon as the deformity stops responding to conservative treatments. The parents of children younger than 10 years of age with non-idiopathic deformities should be informed preoperatively that the deformity may be prone to correction failure or screw loosening after eight-Plate implantation.

The evolutionary transition from outcrossing to selfing can have important genomic consequences. Decreased effective population size and the reduced efficacy of selection are predicted to play an important role in the molecular evolution of the genomes of selfing species. We investigated evidence for molecular signatures of the genomic selfing syndrome using 66 species of Primula including distylous (outcrossing) and derived homostylous (selfing) taxa. We complemented our comparative analysis with a microevolutionary study of P. chungensis, which is polymorphic for mating system and consists of both distylous and homostylous populations. We generated chloroplast and nuclear genomic data sets for distylous, homostylous, and distylous-homostylous species and identified patterns of nonsynonymous to synonymous divergence (dN/dS) and polymorphism (πN/πS) in species or lineages with contrasting mating systems. Our analysis of coding sequence divergence and polymorphism detected strongly reduced genetic diversity and heterozygosity, decreased efficacy of purifying selection, purging of large-effect deleterious mutations, and lower rates of adaptive evolution in samples from homostylous compared with distylous populations, consistent with theoretical expectations of the genomic selfing syndrome. Our results demonstrate that self-fertilization is a major driver of molecular evolutionary processes with genomic signatures of selfing evident in both old and relatively young homostylous populations., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)

The rate of angular correction (ROAC) is very unpredictable and may be affected by various factors in the treatment of genu valgum and varum by means of guided growth. The purpose of this study was to assess the ROAC in cases from our institution and to identify risk factors associated with the occurrence of lower ROAC.We retrospectively reviewed the chart records of 68 patients undergoing guided growth with figure-eight plate for the correction of genu valgum and varum. Based on the data from these patients, the annual increment of physeal growth was calculated and compared with data from the Anderson chart. The associations between patient characteristics and ROAC were evaluated with the use of univariate logistic regression.The mean rate of femoral angular correction was 10.29 degrees/year, while the mean rate of tibial angular correction was 7.92 degrees/year. In a univariate logistic regression analysis, the variables associated with a higher risk of lower ROAC included non-idiopathic coronal deformity of the knee (odds ratio = 13.58, p  < 0.001) and body weight at or above the 95th percentile for children (odds ratio = 2.69, p  = 0.020).Obesity and non-idiopathic coronal deformity of the knee are risk factors for lower ROAC. It is still uncertain whether severity of deformity, race, and operative procedure have a substantial effect on the rate of correction.Level III evidence.

Polymer-based micromolding has been proposed as an alternative to SU-8 micromolding for microfluidic chip fabrication. However, surface defects such as milling marks may result in rough microchannels and micromolds, limiting microfluidic device performance. Therefore, we use chemical and mechanical methods for polishing polymer microchannels and micromolds. In addition, we evaluated their performance in terms of removing the machining (milling) marks on polymer microchannel and micromold surfaces. For chemical polishing, we use solvent evaporation to polish the sample surfaces. For mechanical polishing, wool felt polishing bits with an abrasive agent were employed to polish the sample surfaces. Chemical polishing reduced surface roughness from 0.38 μm (0 min, after milling) to 0.13 μm after 6 min of evaporation time. Mechanical polishing reduced surface roughness from 0.38 to 0.165 μm (optimal pressing length: 0.3 mm). As polishing causes abrasion, we evaluated sample geometry loss after polishing. Mechanically and chemically polished micromolds had optimal micromold distortion percentages of 1.01% ± 0.76% and 1.10% ± 0.80%, respectively. Compared to chemical polishing, mechanical polishing could better maintain the geometric integrity since it is locally polished by computer numerical control (CNC) miller. Using these surface polishing methods with optimized parameters, polymer micromolds and microchannels can be rapidly produced for polydimethylsiloxane (PDMS) casting and thermoplastic hot embossing. In addition, low-quantity (15 times) polymer microchannel replication is demonstrated in this paper.

Objective: Tension band plating has recently gained widespread acceptance as a method of correcting angular limb deformities in skeletally immature patients. We examined the role of biomechanics in procedural failure and devised a new method of reducing the rate of implant failure., Methods: In the biomechanical model, afterload (static or cyclic) was applied to each specimen. The residual stress of the screw combined with different screw sizes and configurations were measured and compared by X-ray diffraction. With regard to static load and similar conditions, the stress distribution was analyzed according to a three-dimensional finite element model., Results: The residual stress was close to zero in the static tension group, whereas it was very high in the cyclic load group. The residual stress of screws was significantly lower in the convergent group and parallel group than in the divergent group. The finite element model showed similar results., Conclusions: In both the finite element analysis and biomechanical tests, the maximum stress of the screw was concentrated at the position where the screws enter the cortex. Cyclic loading is the primary cause of implant failure.

Debate continues regarding the need for a booster vaccination in children who received a universal infant hepatitis B virus (HBV) vaccination. The aim was to explore the need and the strategies for the booster HBV vaccination. 8-year prospective cohort study was conducted among children aged 5-15 years in 2009-2010 in Zhejiang Province. The participants were divided into groups A (<0.1 mIU/mL), B (0.1 to < 1 mIU/mL) and C (1 to <10 mIU/mL) according to the pre-booster anti-HBs antibody levels. 5 μg (group I), 10 μg (group II), 20 μg hepatitis B vaccines (group III) or 5 μg hepatitis A and B (HAB) vaccines (group IV) with 0-1-6-month schedule were randomly administered to children negative for all markers. Blood samples were collected at baseline HBV marker testing, 1 month after the first dose, 1 month, 1 year, 5 years and 8 years after the third dose. Among 4170 children, 2326 (55.8%) were negative for all HBV markers. Group II showed the highest seropositive rates of 92.8%, 99.7%, 97.6%, 90.3% and 83.4% with GMTs of 4194.5 mIU/ml, 4163.9 mIU/ml, 466.9 mIU/ml, 190.6 mIU/ml, 122.6 mIU/ml from 1 month after dose 1 to 8 years after dose 3, respectively ( P < .01). Participants in group C showed seropositive rates of 98.9%, 99.9%, 99.5%, 95.5%, 92.8% after the revaccination with GMTs of 6519.6 mIU/ml, 5267.4 mIU/ml, 547.1 mIU/ml, 249.5 mIU/ml, 155.3 mIU/ml, respectively, higher than group A and B ( P < .001), except 1 month after the third dose. The 10 μg of HBV vaccine with a 0-1-6-month booster regimen may elicit robust responses and persist for 8 years or longer. Additionally, 1-dose revaccination maybe suitable for children with 1 to < 10 mIU/ml anti-HBs titers.

Purpose: Moesin-ezrin-radixin-like protein (Merlin) has been identified as a tumor suppressor in several types of cancers. However, the biological function of Merlin in osteosarcoma remains unclear. MicroRNAs (miRNAs) can influence cancer progression by targeting oncogenes or anti-oncogenes. In this study, we sought to evaluate the regulation of Merlin expression by miR-25-3p and the role of the miR-25-3p/Merlin axis in osteosarcoma progression, with the aim of identifying a potential therapeutic target for osteosarcoma., Materials and Methods: TCGA (The Cancer Genome Atlas) database was used to analyze the correlation between Merlin expression and prognosis. RT-qPCR and Western blotting analyses were performed to compare Merlin expression between normal and malignant cells. A dual-luciferase reporter assay was performed to evaluate the direct targeting of Merlin by miR-25-3p. We overexpressed miR-25-3p, or/and Merlin, in U-2 OS and 143B cells, and studied their cellular functions in vitro. MTT and colony formation assays were performed to determine the effects on cell growth. EdU and cell cycle assays were performed to analyze the effects in cell replication. We used annexin V-fluorescein isothiocyanate and propidium iodide to stain apoptotic cells, and analyzed the cells using flow cytometry. The effects on cell metastasis were studied in wound healing and transwell assays. Lastly, the underlying mechanism was determined in RT-qPCR and Western blotting experiments., Results: Low Merlin expression was linked to poor prognosis. miR-25-3p was observed to directly target Merlin and downregulate its expression. miR-25-3p promoted cell growth, migration, and invasion, and inhibited apoptosis induced by cisplatin. Moreover, the overexpression of Merlin reversed the abovementioned effects of miR-25-3p. Further, the miR-25-3p/Merlin axis was observed to play an important role in the Hippo pathway, and regulated the expression of genes such as BIRC5, CTGF, and CYR61., Conclusion: miR-25-3p functions as an oncogenic microRNA in osteosarcoma by targeting Merlin, and may serve as a potential therapeutic target for osteosarcoma., Competing Interests: The authors declare that they have no conflicts of interest for this work., (© 2020 Rao et al.)

Background: Pediatric femoral neck fracture is a rare injury but yields frequent complications. However, there is a paucity of data regarding the risk factors for these complications., Purpose: The present article reports the rate of complications after femoral neck fracture in pediatric patients and investigates the possible risk factors., Methods: We retrospectively reviewed 44 children (mean age of 9.0 years, range from 2 to 14 years) who were surgically treated for femoral neck fracture in a single trauma center with a mean follow-up of 57.75 months (range from 11 to 224 months). Related clinical factors were recorded and analyzed by multivariable logistic regression., Results: Fracture displacement or Delbet-type fracture had no relation to the injury mechanism. However, younger children experienced severe trauma, combined injury, and low fracture location more than older individuals did. Children with combined injuries were more likely to have a longer waiting time for surgical reduction. Common complications included avascular necrosis (AVN) in 14 cases, nonunion of fracture in 2 cases, coxa vara in 4 cases, and premature physeal closure (PPC) in 7 cases. Only the Delbet type was an independent predictor of AVN (OR = 0.14, p = 0.030). Inadequate reduction was associated with higher rates of coxa vara (OR = 33.19, p = 0.032). Epiphysis penetration in children younger than 10 years old increased the rate of PPC (p = 0.032). No significant risk factor was found for fracture nonunion., Conclusion: For femoral neck fracture in pediatric patients, both the injury mechanism and fracture characteristics have age-related distributions. Early reduction should be carried out as early as possible based on the safe condition of the child, but for younger children, transepiphyseal fixation should be avoided. AVN may be intrinsic to injury characteristics rather than resulting from the choice of treatment mode.

Background: The purpose of this study was to evaluate the effect, rate of angular correction, and complications of temporary hemiepiphysiodesis (TH) in the treatment of skeleton immature posttraumatic genu angular deformity., Methods: We retrospectively reviewed the records of 27 patients undergoing temporary hemiepiphysiodesis for the management of posttraumatic genu angular deformity. Based on the data from these patients, the rate of correction, effect of correction, length of the lower limbs, and complications were used as the outcome measures., Results: Outcome measurements were obtained from a chart review of medical records that included information about clinical evaluations. Fifteen boys and 12 girls, with an average age of 6.3 years, were included in the study. The average follow-up was 3.8 years (range, 1.9 to 5.9 years) after surgery. Complete correction was obtained in 24 patients, while partial correction was obtained in 3 patients. The mean rate of angular correction was 8.41°/year in distal femur and 15.19°/year in proximal tibia. One patient had recurrence of genu valgum. No leg length discrepancy was found in our patients., Conclusion: Temporary hemiepiphysiodesis is a simple, effective, reliable, and reproducible method for the treatment of posttraumatic genu angular deformity, with fewer complications than osteotomy. Nevertheless, it is important to follow the rebound patient closely until skeletal maturity in our future work.

Evolutionary transitions from outcrossing to selfing often occur in heterostylous plants. Selfing homostyles originate within distylous populations and frequently evolve to become reproductively isolated species. We investigated this process in 10 species of Primula section Obconicolisteri using phylogenomic approaches and inferred how often homostyly originated from distyly and its consequences for population genetic diversity and floral trait evolution. We estimated phylogenetic relationships and reconstructed character evolution using the whole plastome comprised of 76 protein-coding genes. To investigate mating patterns and genetic diversity we screened 15 microsatellite loci in 40 populations. We compared floral traits among distylous and homostylous populations to determine how phenotypically differentiated homostyles were from their distylous ancestors. Section Obconicolisteri was monophyletic and we estimated multiple independent transitions from distyly to homostyly. High selfing rates characterised homostylous populations and this was associated with reduced genetic diversity. Flower size and pollen production were reduced in homostylous populations, but pollen size was significantly larger in some homostyles than in distylous morphs. Repeated transitions to selfing in section Obconicolisteri are likely to have been fostered by the complex montane environments that species occupy. Unsatisfactory pollinator service is likely to have promoted reproductive assurance in homostyles leading to subsequent population divergence through isolation., (© 2019 The Authors. New Phytologist © 2019 New Phytologist Trust.)

Analysis of population genetics provides insights into the evolutionary processes, among which the sample size choice is per se a crucial issue in the analysis. Genome-wide high-throughput techniques based on RADseq have been increasingly used in studies on the population genomics of invasive species. However, there is little information available regarding optimal sample sizes for analyzing population genomics of invasive species. In this study, we first use type IIB endonucleases restriction site-associated DNA (2b-RAD) to mine thousands of single nucleotide polymorphisms (SNPs) for native and introduced populations in Q1 clade (SPB and 17JN) and Q2 clade (ISQ and UAS0601) of the whitefly, Bemisia tabaci (Gennadius) MED (also known as B. tabaci biotype Q). Then, we used resampling techniques to create simulated populations with a random subset of individuals and 3,000 SNPs to determine how many individuals should be sampled for accurate estimates of intra- and interpopulation genetic diversity. We calculated the intrapopulation genetic diversity parameters (unbiased expected heterozygosity, observed heterozygosity, and the number of effect alleles) and pairwise genetic differentiation F ST ; finally, an ad hoc statistic, Δ K , was used to determine the optimal value. Our results showed that a sample size greater than four individuals ( n  ≥ 4) has little impact on estimates of genetic diversity within whitefly populations; moreover, precise estimate of F ST can be easily achieved at a very small simple size ( n  = 3 or 4). Our results will provide in-depth understanding of the optimization of sampling schemes in population genomics of invasive species., Competing Interests: None declared., (© 2019 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Objective: To investigate the awareness of preconception care among women of child-bearing age with type 1 diabetes (T1DM) and their self-management status, in order to provide evidence for establishment of management pathway for women with T1DM in pregnancy in China. Methods: This cross-sectional survey recruited female participants of child-bearing age from the cohort of Guangdong Type 1 Diabetes Translational Medicine Study conducted between June 2011 and December 2017. The participants were asked to fill out a questionnaire on the awareness of preconception care, their frequency of self-monitoring of blood glucose (SMBG) and other related variables. Chi-squared test or chi-squared test for trend was used in comparisons of categorical variables, and logistic regression analysis was performed to assess associated factors. Results: Totally, 441 women of child-bearing age with T1DM were investigated. The results show that their awareness of preconception care was poor (15.42%, 68/441). Higher educational level (χ(2trend)=3.990, P= 0.046), experience of post-diabetes education evaluation ( P< 0.001), and better coverage of different modules in diabetes education (survival skills: χ(2)=7.525, P= 0.004; basic knowledge: χ(2)=8.598, P= 0.002; advanced knowledge: P< 0.001) were associated with better awareness of preconception care. The average frequency of SMBG in these participants was 0.29 (0.14, 2.00) times per day, and only 8.5% (37/435) of them reached the frequency (≥4 times per day) recommended by guidelines. Moreover, 21.1% (92/435) of them hardly ever performed SMBG. Conclusion: Child-bearing age women with T1DM in Gunangdong had poor awareness of preconception care, with a much lower SMBG frequency than recommendation.

Ceropegia jinshaensis D.T.Liu & Z.K.Wu (Asclepiadoideae, Apocynaceae), a new species from northwestern Yunnan along the upper Yangtze river of China, is described and illustrated. This species is similar to C. meleagris H. Huber, C. dorjei C. E. C. Fischer and C. aridicola W. W. Smith, but can be distinguished easily by its leaf shape and floral features, especially the corolla shape and size, the interior of corolla tube and coronal characters.

Primula dongchuanensis Z.K.Wu & Yuan Huang, a new species of Primulaceae from Dongchuan of northern Yunnan, China, is described and illustrated. Both morphological and molecular evidence support P. dongchuanensis as a member of the sect. Proliferae. It is similar to P. aurantiaca W.W.Smith & Forrest, but is distinguished by having unique raceme inflorescences. Its distribution, phenology and conservation status are also provided.

Primula filchnerae (Primulaceae) is an endangered endemic herb species in China. In this study, we characterized the complete chloroplast genome of P. filchnerae based on next generation sequencing (NGS). The chloroplast genome of P. filchnerae was 151,547 bp in size, containing a large single-copy (LSC) region of 82, 662 bp and a small single-copy (SSC) region of 17,749 bp. These two regions were separated by a pair of inverted repeat regions (IRs), each of 25,568 bp. A total of 130 functional genes were encoded, consisted of 86 protein-coding genes, 36 tRNA genes, and eight rRNA genes., Competing Interests: No potential conflict of interest was reported by the authors., (© 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Objective: To examine the clinical effects of Yisui Shengxue Granules () in the treatment of β-thalassemia and explore its mechanism on DNA methylation levels., Methods: A randomized placebo-controlled double-blinded trial was conducted. Forty patients with β-thalassemia were recruited and distributed randomly by envelope method into an experimental group and a control group, 20 patients in each group. The patients were given Yisui Shengxue Granules in the experimental group and placebo in the control group (12 g/bag three times a day) during a 3-month intervention. Before and after 1, 2, and 3 months of treatment, peripheral intravenous blood was sampled, and blood parameters such as hemoglobin (Hb), red blood cells (RBCs), reticulocytes (Ret), and fetal hemoglobin (HbF) were analyzed. Mononuclear cells from 5 patients, who showed an obvious treatment effect, were isolated by density gradient centrifugation. DNA methylation was analyzed using an Affymetrix USA GeneChip Human Promoter 1.0 Array and Input-promoter 1.0., Results: Compared with pre-treatment, there was an obvious increase in Hb and RBCs counts after 1, 2, and 3 months in the experiment group (P<0.01 or P<0.05). Meanwhile, HbF increased from the 2nd to the 3rd month (P<0.05). In the control group, Hb and RBCs showed no obvioas change. After 3-month treatment, DNA methylation results from 5 patients revealed that there were 24 hypomethylated genes and 3,685 hypermethylated genes compared with pre-treatment. Genes of insulin-like growth factor 1 receptor (IGF1R) and Janus kinase 3 (JAK3) revealed the most relations with other genes (degree: 21) and genes of 1-phosphatidylinositol-4, 5-bisphosphate phosphodiesterase gamma 2 (PLCG2) and mitogen-activated protein kinase 10 (MAPK10) showed a stronger intermediary role (betweenness centrality=0.04)., Conclusions: JAK3 and MAPK10 are two key genes in bone marrow and the lymphatic system, and JAK3 is likely to be related to hematopoietic cytokines in the process of early hematopoiesis. (Registration No. NCT01549080).

Pancreatic endocrine islets are vital for glucose homeostasis. However, the islet developmental trajectory and its regulatory network are not well understood. To define the features of these specification and differentiation processes, we isolated individual islet cells from TgBAC(neurod1:EGFP) transgenic zebrafish and analyzed islet developmental dynamics across four different embryonic stages using a single-cell RNA-seq strategy. We identified proliferative endocrine progenitors, which could be further categorized by different cell cycle phases with the G1/S subpopulation displaying a distinct differentiation potential. We identified endocrine precursors, a heterogeneous intermediate-state population consisting of lineage-primed alpha, beta and delta cells that were characterized by the expression of lineage-specific transcription factors and relatively low expression of terminally differentiation markers. The terminally differentiated alpha, beta, and delta cells displayed stage-dependent differentiation states, which were related to their functional maturation. Our data unveiled distinct states, events and molecular features during the islet developmental transition, and provided resources to comprehensively understand the lineage hierarchy of islet development at the single-cell level., (© The Author(s) (2019). Published by Oxford University Press on behalf of Journal of Molecular Cell Biology, IBCB, SIBS, CAS.)