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Kernicterus is a severe complication of extreme neonatal hyperbilirubinemia. Prolonged exposure to high-level unconjugated bilirubin (UCB) directly damages brain tissue. Neuroinflammation is believed to contribute to UCB-induced neurotoxicity. Pyroptosis has been as a highly inflammatory form of programmed cell death. Therefore, this study aimed to explore whether pyroptosis was involved in the pathogenesis of UCB neurotoxicity in kernicterus model rats. VX-765, a specific inhibitor of caspase-1, was intraperitoneally administered to the model rats to observe its effects on the short-term and long-term outcomes of the model animals at the molecular, cellular, morphological, and behavioral levels. The results indicated that UCB significantly induced the activation of caspase-1 and gasdermin D(GSDMD), and VX-765 inhibited caspase-1-GSDMD pathway. Compared with those of the UCB group and the vehicle+UCB group, VX-765-treated rats released lower levels of IL-1β and IL-18. Furthermore, H&E and TUNEL staining showed that nerve cells in the VX-765-treated group were better preserved and had less DNA fragmentation. Most importantly, VX-765 improved both the short-term and long-term neurological functions of kernicterus model rats. This study demonstrated that pyroptosis was involved in the pathogenesis of kernicterus through caspase-1 activation, which could be inhibited by VX-765, exerting a neuroprotective effect in kernicterus model rats.

To explore the spatial distribution and intraspecific correlation of为探究锐齿槲栎种群的空间分布特征及关联性,本研究以秦岭皇冠暖温性落叶阔叶林25 hm

In this study, the spatial distribution patterns of individuals with different diameter classes of dominant shrub以秦岭皇冠暖温性落叶阔叶林25 hm

Kernel development plays an important role in determining kernel size in maize. Here we present the cloning and characterization of a maize gene, nitrate transporter1.5 (NRT1.5), which controls small kernel phenotype by playing an important role in kernel development. A novel recessive small kernel mutant miniature2-m1 (mn2-m1) was isolated from self-pollinated progenies of breeding materials. The mutant spontaneously showed small kernel character arresting both embryo and endosperm development at an early stage after pollination. Utilizing 21 polymorphic SSR markers, the mn2-m1 locus was limited to a 209.9-kb interval using 9 176 recessive individuals of a BC1 segregating population from mn2-m1/B73. Only one annotated gene was located in this 209.9 kb region, Zm00001d019294, which was predicted to encode nitrate transporter1.5 (NRT1.5). Allelism tests confirmed that mn2-m1 was allelic to miniature2-m2 (mn2-m2) and miniature2-710B (mn2-710B). The mn2-m1 and mn2-m2 alleles both had nucleotide deletions in the coding region resulting in premature termination, and the mn2-710B allele had some missence mutations. Subcellular localization showed that Miniature 2 (MN2) is localized in the plasma membrane. Quantitative real-time PCR (qRT-PCR) analysis revealed that the expression of MN2 and some genes involved in the basal endosperm transfer layer (BETL) and embryo surrounding region (ESR) development were affected in mn2-m1 seeds. These results suggested that MN2 plays an important role in maize seed development.

The Qinling Mountain is a natural boundary between warm temperate zone and subtropical zone. While the China Forest Biodiversity Monitoring Network (CForBio) have basically covered most of the climate regions in China, few plots were located in the climate transition zone. Following the field protocol of CForBio and the Center for Tropical Forest Science (CTFS), a 25 hm秦岭是典型的暖温带-亚热带过渡区。中国森林生物多样性监测网络(CForBio)现有的样地已经基本覆盖了中国大多数气候区,但位于气候过渡带的样地较少。本研究参照CForBio和CTFS(Center for Tropical Forest Science)的技术规范,于2019年在陕西皇冠山省级自然保护区内建立25 hm

AIM: To evaluate the therapeutic effect of amniotic membrane (AM) for covering high myopic macular hole associated with retinal detachment following failed primary surgery. METHODS: Seventeen eyes of 17 patients whose axial length was more than 29 mm suffered from macular hole (MH) or MH associated with retinal detachment (RD), and had previously surgery of pars plana vitrectomy (PPV) with internal limiting membrane (ILM) peeling and silicone oil (SO) tamponade. Half a year after the surgery, optical coherence tomography (OCT) showed that MH did not heal in all 17 eyes and RD was still maintained in 13 eyes of these 17 eyes. We performed SO removal combined with AM covering on macular area and C3F8 tamponade, and phacoemulsification combined with intraocular lens implantation simultaneously cataract eyes. We followed up these patients for one year. RESULTS: In all 17 eyes, SO was removed successfully, MHs were healed and RDs were reattached. One eye (5.89%, 1/17) had AM shifted half a month after surgery and underwent a second surgery to adjust the position of the AM and supplement C3F8. After surgery, the visual acuity (VA) improved in 15 eyes (88.24%, 15/17), no change in two eyes (11.76%, 2/17). No serious complications occurred in all eyes. CONCLUSION: AM covering is helpful to rescue the previous failure surgery of high myopic MH.

OBJECTIVE: To study whether pyroptosis is involved in the bilirubin-induced injury of primary cultured rat cortical microglial cells. METHODS: Primary cultured rat cortical microglial cells were randomly administered with 30 μmol/L bilirubin (bilirubin group), 30 μmol/L bilirubin following 30 μmol/L VX-765 pretreatment (VX-765+bilirubin group), or an equal volume of dimethyl sulfoxide (control group). Modified MTT assay was used to measure the viability of microglial cells. Western blot was used to measure the expression of the pyroptosis-related proteins Caspase-1 and gasdermin D (GSDMD). Lactate dehydrogenase (LDH)-release assay was used to evaluate the cytotoxicity of microglial cells. EtBr/EthD2 with different molecular weights (394 Da/1 293 Da) was used to measure the size of plasma membrane pores. ELISA was used to measure the level of the inflammatory factor interleukin-1β (IL-1β) in culture supernatant. RESULTS: After bilirubin stimulation, the viability of microglial cells decreased and LDH release increased, both in a time-dependent manner. Compared with the control group, the bilirubin group had a significantly higher positive rate of small-molecule EtBr passing through the cell membrane (P < 0.001), while there was no significant difference in the pass rate of large-molecule EthD2 between groups (P > 0.05). The expression of activated Caspase-1 significantly increased at 0.5 hour after bilirubin stimulation (P < 0.05), and that of activated GSDMD significantly increased at 6 hours after bilirubin stimulation (P < 0.05). The release of IL-1β significantly increased at 6 hours after bilirubin stimulation and reached the peak at 24 hours (P < 0.001). Compared with the bilirubin group, the VX-765+bilirubin group had a significant increase in cell viability (P < 0.05) and significant reductions in the expression of activated GSDMD, the pass rate of EtBr, and the release of LDH and IL-1β (P < 0.05). CONCLUSIONS: Pyroptosis is involved in bilirubin-induced injury of primary cultured microglial cells.

The study aimed to evaluate the protective effect of Dendrobium officinale flower extraction (DOFE) on alcohol-induced liver injury and its probable mechanisms in mice. The chemical composition of DOFE was performed via UPLC/MS. Male Kunming mice were used to establish alcohol-induced liver injury models by oral gavage of 56% alcohol. Results showed that DOFE dramatically attenuated the increased serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), and triacylglycerol (TG). Meanwhile, hematoxylin and eosin and Oil Red O staining showed that DOFE attenuated degeneration, inflammatory infiltration, and lipid droplet accumulation. DOFE was also found to suppress the activity of malonaldehyde (MDA) and enhanced the level of glutathione (GSH) and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) in the liver. The protection of DOFE against oxidative stress was associated with the downregulation of hepatic cytochrome P450 2E1 (CYP2E1) and upregulation of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and NAD(P)H quinone oxidoreductase l (NQO1). Additionally, DOFE suppressed inflammation via downregulating Toll-like receptor-4 (TLR-4) and nuclear factor kappa-B P65 (NF-κB P65). Thus, DOFE exhibited a significant protective effect against alcohol-induced liver injury through its antisteatosis, antioxidative, and anti-inflammatory effect.

An effective classifier combining convolutional neural network and regularized extreme learning machine (called as CNN-RELM) is presented in this paper. Firstly, CNN-RELM trains the convolutional neural network (CNN) using the gradient descent method until the learning target accuracy reaches. Then the fully connected layer of CNN is replaced by regularized extreme learning machine (RELM) optimized by genetic algorithm and the rest layers of the CNN remain unchanged. The experiments on different face databases are given to evaluate the performance of CNN-RELM. The experimental results show that CNN-RELM is a feasible classifier and it outperforms CNN and RELM. Due to the uniting of CNN and RELM, CNN-RELM have the advantages of CNN and RELM and it is easier to learn and faster in testing.

Bilirubin encephalopathy, the most serious complication of hyperbilirubinemia during the neonatal period, with high mortality and morbidity, often causes irreversible neurological damage. Currently, caspase-1, a member of the cysteinyl aspartate-specific protease caspase family, is regarded as a key mediator of inflammatory processes, attracting widespread attention. The purpose of this study was to investigate whether caspase-1 is involved in bilirubin-induced neuronal injury. VX-765, a highly potent and selective inhibitor of caspase-1, was used to investigate the effects of unconjugated bilirubin (UCB) on rat cortical neurons, including cell viability, morphological changes in the cell membrane, and nuclear factor-kappa B (NF-κB) activation. Neurons treated with UCB showed increased caspase-1 activity without the secretion of interleukin (IL)-1β and IL-18, and caspase-1 was significantly inhibited by pretreatment with VX-765. The cell viability of the VX-765-pretreated neurons was improved, and cell membrane rupture was prevented, as detected by lactate dehydrogenase release and ethidium bromide uptake. Moreover, NF-κB activation by UCB exposure, was attenuated by VX-765 pretreatment. Bilirubin-induced neuronal injury involves the activation of caspase-1 and NF-κB, leading to membrane leakage, independently of IL-1β and IL-18.

Davalliaceae is a small family of derived ferns that grow epiphytic or occasionally epilithic in the tropics and subtropics of the Old World. Despite recent taxonomic and phylogenetic achievements, some aspects of its classification remain controversial. In this study, we provided new spore observations on Davalliaceae by using scanning electron microscopy. The newly generated data were integrated together with previously reported spore evidence of this group. The evolution of spore ornamentation and taxonomic implications of spore characters was optimized onto a phylogenetic tree based on plastid sequences. Verrucate colliculate spore ornamentation was identified as plesiomorphic character state within Davalliaceae. The Humata clade was characterized by the transition from verrucate colliculate to fused verrucate ornamentation. Several species complexes subject to taxonomic controversy were found to show informative variation in the spore ornamentation. In conclusion, our observations support the utility of spores as a morphological taxonomic trait in ferns, as long as the evolution of spore morphology is considered.

Although the phylogenetic framework of Davalliaceae is known, the classification of Chinese Davalliaceae is still controversial. In this study, a molecular phylogenetic tree of 60 accessions, including 29 species produced in China, was constructed using five plastid DNA markers-atpB, atpB-rbcL, rbcL, rbcL-accD, and accD. New data on studied specimens, field investigations, and scanning electron microscopy analysis of leaf epidermis and spores were used to reclassify Chinese Davalliaceae. The taxonomic position of Davallia canariensis was confirmed based on new evidence and a new key to sections of Chinese Davalliaceae was proposed. The taxonomically controversial genus Paradavallodes was confirmed as a polyphyletic group, and it was assigned to Davallia sect. Trogostolon and Davallia sect. Davallodes. Further, species endemic to China were delimited, 21 species were admitted to six sections of Davallia, two new combinations were proposed, two new synonyms were defined and a new key to Chinese species of Davalliaceae was presented.

A method based on quantum genetic algorithm (QGA) is presented to train and implement morphological neural network (MNN) in this paper. The algorithm adjusts the weights and biases of MNN and the QGA automatically determines the learning rate of the algorithm. Then the trained MNN is applied to image restoration. The image restoration simulation and a comparison with the median filter are shown in the end. We can see that the MNN is a quite good method applied to image restoration.

Two consecutive years (4-crop) experiments were conducted to study the influence of different paddy rice-upland crop rotation in cold-waterlogged paddy field on the growth of crops and soil characteristics. The result showed that compared with the rice-winter fallow (CK) pattern, the two-year average yield of paddy rice under four rotation modes, including rape-rice (R-R), spring corn-rice (C-R), Chinese milk vetch-rice (M-R) and bean-rice (B-R), were increased by 5.3%-26.7%, with significant difference observed in C-R and R-R patterns. Except for M-R pattern, the annual average total economic benefits were improved by 79.0%-392.4% in all rotation pattern compared with the CK, and the ration of output/input was enhanced by 0.06-0.72 unit, with the most significant effect found in the C-R pattern. Likewise, compared with the CK, the contents of chlorophyll and carotenoid, and net photosynthetic rate (Pn) of rice plant were all increased during the full-tillering stage of rice in all rotation patterns. The rusty lines and rusty spots of soils were more obvious compared with the CK during the rice harvest, particularly in R-R, C-R and B-R patterns. The ratio of water-stable soil macro aggregates of plough layer of soil (2 mm) decreased at different levels in all rotation patterns while the ratios of middle aggregate (0.25-2 mm, expect for M-R) and micro aggregate of soil (0.25 mm) were opposite. There was a decreasing trend for soil active reducing agents in all rotation patterns, whereas the available nutrient increased. The amounts of soil bacteria in C-R and B-R patterns, fungi in B-R rotation pattern, cellulose bacteria in R-R, C-R and B-R patterns and N-fixing bacteria in B-R pattern were improved by 285.7%-403.0%, 221.7%, 64.6-92.2% and 162.2%, respectively. Moreover, the differences in all microorganisms were significant. Thus, based on the experimental results of cold-waterlogged paddy field, it was concluded that changing from single cropping rice system to C-R, R-R and B-R rotation patterns had good effect in terms of improving total yield and economic benefits, and soil physical and chemical properties were improved.

The yields of cold-waterlogged (CW) paddy fields widely spreading in Jiangnan mountainous areas are moderate or low but have a high potential to be increased. Based on data including 41 soil characteristics of 17 pairs of typical surface soils of cold-waterlogged paddy field and non cold-waterlogged (NCW) paddy field at a neighboring landscape unit in Fujian Province, various index differences of soil properties and causes between CW paddy field and NCW paddy field were systematically studied, and a minimum data set (MDS) of soil quality assessment for CW paddy field was established by principal component analysis. By pair analysis, soil characteristics of CW paddy field showed that the content of organic matter increased by 31.7%, but the microbial biomass C decreased by 37.8%, which belonged to active soil organic matter component. The content of ferrous iron (Fe2+) increased by 177.0%, but the available phosphorus (P) and potassium (K) decreased by 52.3% and 22.8%, respectively. Catalase and invertase activities increased by 58.3% and 22. 1%, but phosphatase, nitrate reductase activities and microflora decreased by 47. 8%, 66.6% and 29.8%-46.0%, respectively. The sand content increased about 8.0%, but the water immersed bulk density decreased by 25.8%. There were significant differences of indices for 28 of all 41 soil characteristics. Five principal components cumulatively exhibiting about 78.5% contribution were concluded from the 28 soil characteristics to reflect characteristics related to soil biochemistry, active organic nitrogen, reducing barriers, physical and chemical nutrients, respectively. Eventually, correlation analysis combined with expert experience method were applied to optimize MDS containing six factors for soil quality assessments, including C/N, bacteria, microbial biomass N, total reducing agents, physical sand and total P.

甜菜碱是细胞内重要的渗透调节物质，其积累能有效提高植物对非生物逆境的抗性。以转 betA 基因小麦和未转基因的野生型为材料，在灌浆期模拟干热风胁迫处理植株3 d，研究干热风对植株生长和籽粒产量的影响。胁迫处理后，转基因植株保持较好长势，叶片青枯失水较少，旗叶持绿面积显著大于野生型。各株系植株的光合作用能力和蔗糖磷酸合成酶(SPS)及蔗糖合成酶(SS)活性在胁迫处理后都下降，转基因植株的净光合速率下降到2.3~3.7 μmol CO 2 m -2 s -1 ，而野生型下降到1.2 μmol CO 2 m -2 s -1 ；野生型植株的SPS和SS活性的下降幅度分别为处理前56.8%和53.9%, 而转基因株系的下降幅度分别为62.3%~69.8%和56.5%~64.5%。干热风胁迫使得各株系植株的旗叶甜菜碱含量升高，但转基因株系的叶片甜菜碱含量比野生型的高18%~87%。在甜菜碱保护作用下，转基因植株在胁迫条件下能够维持较高的光合速率，合成较多的碳水化合物，使得百粒重和单株产量均高于野生型。因此，通过转 betA 基因可显著提高小麦在干热风处理条件下的甜菜碱含量从而增强其抗干热风能力。

In this paper, we unveil a novel naphthalimide-porphyrin hybrid based fluorescence probe (1) for ratiometric detection of Hg(2+) in aqueous solution and living cells. The ratiometric signal change of the probe is based on a carefully predesigned molecule containing two independent Hg(2+)-sensitive fluorophores with their maximal excitation wavelengths located at the same range, which shows reversibly specific ratiometric fluorescence responses induced by Hg(2+). In the new developed sensing system, the emissions of the two fluorophores are well-resolved with a 125 nm difference between two emission maxima, which can avoid the emission spectra overlap problem generally met by spectra-shift type probes and is especially favorable for ratiometric imaging intracellular Hg(2+). It also benefits from a large range of emission ratios and thereby a high sensitivity for Hg(2+) detection. Under optimized experimental conditions, the probe exhibits a stable response for Hg(2+) over a concentration range from 1.0 x 10(-7) to 5.0 x 10(-5) M, with a detection limit of 2.0 x 10(-8) M. The response of the probe toward Hg(2+) is reversible and fast (response time less than 2 min). Most importantly, the ratiometric fluorescence changes of the probe are remarkably specific for Hg(2+) in the presence of other abundant cellular metal ions (i.e., Na(+), K(+), Mg(2+), and Ca(2+)), essential transition metal ions in cells (such as Zn(2+), Fe(3+), Fe(2+), Cu(2+), Mn(2+), Co(2+), and Ni(2+)), and environmentally relevant heavy metal ions (Ag(+), Pb(2+), Cr(3+), and Cd(2+)), which meets the selective requirements for biomedical and environmental monitoring application. The recovery test of Hg(2+) in real water samples demonstrates the feasibility of the designed sensing system for Hg(2+) assay in practical samples. It has also been used for ratiometric imaging of Hg(2+) in living cells with satisfying resolution, which indicates that our novel designed probe has effectively avoided the general emission spectra overlap problem of other ratiometric probes.

To study the chemical constituents from the capitula of Eriocaulon australe.The compounds were isolated and purified by column chromatography and their structures were determined through spectroscopic techniques (NMR)and physicochemical properties.Six compounds were identified from capitula of Eriocaulon australe as (R)-semixanthomegnin( 1) ,tora-lactone-9-0-β-D-glucopyranoside(2), ( - ) -semivioxanthin-9-0-β-D-glucopyranoside( 3) ,4-ketopinoresinol(4) ,β-daucosterol (5) and 3,3'-dihydroxy-4,4'-dimethoxybiphenyl(6).Compounds 1 ~ 6 are isolated from this plant and compounds 3 ~ 6 are ob- tained from Eriocaulon genus for the first time. Compound 1 shows DNA scission activity.

Objective: To investigate whether LCRG1 (Laryngeal Carcinoma Related Gene 1) has tumor suppressor function. Methods: The recombinant plasmid pcDNA3.1(+)/LCRG1 was successfully constructed. The biological effects of LCRG1 on Hep-2 cell line were studied by cell transfection, cell growth observation colony formation analysis and tumorigenicity experiments. Results: The LCRG1 gene potently inhibited tumorgenesis in vitro and in vivo, as showed by dramatic growth arrest observed in cell growth analysis and suppression of anchorage-independent growth and tumorigenicity in nude mice. Conclusion: Our results suggested that LCRG1 may be a candidate of tumor suppressor gene.

Guangdong Province, located in southern China, is of great interest in studies of plant geography and conservation in limestone areas because of its rich biodiversity. We: (1) performed a phytogeographical analysis of pteridophytes in the limestone areas in Guangdong, (2) evaluated and determined levels of endemism, and (3) identified affinities with the floras of neighboring limestone areas. We found that some families show a preference for limestone habitats, e.g., Hypodematiaceae, Lygodiaceae, and Adiantaceae, are typical elements among pteridophytes in the limestone regions of Guangdong. In addition, some genera or section, such as Hypodematium, and Polystichum sect. Cyrtogonellum, show an exclusive preference for limestone habitats. The predominant phytogeographical affinities identified were East Asia, Chinese endemic, and Tropical Asia. We concluded that: (1) the limestone fern flora of Guangdong is principally subtropical in nature and represents southern subtropical flora at the margins of the northern tropics, with a strong interaction with the East Asian flora, (2) the limestone fern flora of Guangdong has a phytogeographical affinity with Japan, though the floras of neighboring Guangxi and Hainan Provinces have higher similarity with it.

This letter described the design and synthesis of a novel fluorescein-appended rhodamine spirolactam derivative and its preliminary application as a ratiometric fluorescent cellular imaging probe for Zn(2+). The ratiometric fluorescent signal change of the probe is based on an intramolecular fluorescence resonance energy transfer (FRET) mechanism modulated by a specific metal ion induced ring-opening process of the rhodamine spirolactam (acting as a trigger). In the new developed sensing system, the emission peaks of the two fluorophores are well-resolved, which can avoid the emission spectra overlap problem generally met by spectra-shift type probes and benefits for observation of fluorescence signal change at two different emission wavelengths with high resolution. It also benefits for a large range of emission ratios, thereby a high sensitivity for Zn(2+)detection. Under optimized experimental conditions, the probe exhibits a stable response for Zn(2+) over a concentration range from 2.0 x 10(-7) to 2.0 x 10(-5) M, with a detection limit of 4.0 x 10(-8) M. Most importantly, the novel probe has well solved the problem of serious interferences from other transition metal ions generally met by previously reported typical fluorescent probes for Zn(2+) with the di(2-picolyl)amine moiety as the receptor (in this case, the fluorescence response induced by Cd(2+)is even comparable to that of Zn(2+)) and shows a reversible and fast response toward Zn(2+). All these unique features make it particularly favorable for ratiometric cellular imaging investigations. It has been preliminarily used for ratiometric imaging of Zn(2+) in living cells with satisfying resolution.

Basic principles of clinical trials of nonpharmacologic treatment are similar to those of pharmacologic treatment, but its some special characteristics should be discussed.To explore the design characteristics of clinical surgery trial through the example of tunnel thread-drawing therapy for simple anal fistula.The clinical trial was designed as a prospective, controlled, randomized multicenter trial. The patients came from Longhua Hospital, Yueyang Hospital of Integrated Traditional Chinese and Western medicine, and Affiliated Hospital of Jianxi University of Traditional Chinese Medicine. Stratified random was performed according to the subtype of anal fistula including low and high anal fistula. Full analysis set was applied to analyze the baseline data, and per protocol set was used in efficacy and safety analysis. The intervention was tunnel thread-drawing method. Classical therapy of thread-drawing method was employed as positive control.The primary outcomes were course of recovery and cure rate. Quality of life score was used as secondary outcome and anal maximum constriction pressure was considered as safety outcome.Optimal efficiency testing method was used to estimate sample size. A total of 244 subjects were recruited and 236 subjects completed the trial. There was no significant difference in the cure rate of the low and high anal fistula between the treatment group and control group. There was a significant difference in the course of recovery in the patients with low and high anal fistula between treatment group and control group (P0.01). To the patients with low anal fistula, the course of recovery in the treatment group was (22.26 + or - 8.67) d, and the course of recovery in the control group was (31.41+ or - 11.39) d. To the patients with high anal fistula, the course of recovery in the treatment group was (24.73 + or - 8.15) d, and the course of recovery in the control group was (32.20 + or - 12.60) d. There was no significant difference in the scores of quality of life in the patients with low anal fistula between the treatment group and control group (P0.05). The trial showed that the tunnel thread-drawing therapy was significantly more effective than classical method for improving the anal sphincter function and patient satisfaction with treatment in the patients with high anal fistula. However the other items of quality of life in the two groups did not show significant difference. There was no any adverse event report in each group. There was no significant difference in the anal maximum constriction pressure in the treatment group before and after the operation.The trial shows that the tunnel thread-drawing therapy for simple anal fistula can shorten the course of recovery and improve the patients' quality of life. The training about surgical intervention and clinical implementation program is important in clinical surgery trial. Blind is absolutely difficult to implement and placebo cannot be used in this kind of trial.

To investigate the therapeutic effect of PI3Kgamma inhibitor AS605240 on autoimmune myocarditis in mice.BALB/c mice were randomly divided into three groups, AS605240 group and vehicle group were injected subcutaneously with emulsions containing CFA and 100 ng peptide which derived from murine cardiac alpha-myosin heavy chain on day 0 and 7 while control group were injected with emulsions containing CFA and PBS. AS605240 group received the oral administration of AS605240 50 mg/(kg x d). The vehicle group received the oral administration of an equal volume of 0.5% carboxymethylcellulose. 21 days after the first immunization, mice were sacrificed, heart and body weight were measured. Myocarditis severity was evaluated according to a semi-quantitative scoring system in heart sections. Immunohistochemistry was performed to determine the effect of AS605240 on myocardium macrophage infiltration; TNF-alpha levels in myocardium were determined by ELISA. In vitro and in vivo chemotaxis assays were performed to determine the effect of AS605240 on MCP-1-induced macrophage chemotaxis.Histological examination of the heart showed that AS605240 significantly relieved the murine myocarditis and reduced heart/body weight ratios in experimental autoimmune myocarditis (EAM) (P0.01). Immunohistochemical detection showed that AS605240 significantly suppressed macrophage infiltration into the heart with EAM. ELISA demonstrated that AS605240 down-regulated TNF-alpha levels in myocardium (P0.01). In vitro and in vivo chemotaxis assays indicated that AS605240 significantly suppressed MCP-1-induced macrophage chemotaxis (P0.01).AS605240 may be an effective drug for autoimmune myocarditis, of which the mechanism is relating to suppress macrophage chemotaxis and macrophage infiltration into myocardium, and to decrease TNF-alpha levels in myocardium.

The critical role of phosphoinositide 3-kinase gamma (PI3Kgamma) in inflammatory cell activation and recruitment makes it an attractive target for immunomodulatory therapy. 5-Quinoxilin-6-methylene-1,3-thiazolidine-2,4-dione (AS605240), a potent PI3Kgamma inhibitor, has been reported to ameliorate chronic inflammatory disorders including rheumatoid arthritis, systemic lupus erythematosus, and atherosclerosis. However, its in vivo effect on intestinal inflammation remains unknown. Here we evaluated the protective and therapeutic potentials of AS605240 in mice with dextran sodium sulfate (DSS)-induced acute and chronic colitis. Our results showed that AS605240 improved survival rate, disease activity index, and histological damage score in mice administered DSS in both preventive and therapeutic studies. AS605240 treatment also significantly inhibited the increase in myeloperoxidase levels, macrophage infiltration, and CD4(+) T-cell number in the colon of DSS-fed mice. The DSS-induced overproduction of colonic proinflammatory cytokines including interleukin (IL)-1beta, tumor necrosis factor-alpha, and interferon-gamma was significantly suppressed in mice undergoing AS605240 therapy, whereas colonic anti-inflammatory cytokines such as IL-4 were up-regulated. The down-regulation of the phospho-Akt level in immunological cells from the inflamed colon tissue and spleen of AS605240-treated mice was detected both by immunohistochemical analysis and Western blotting. These findings demonstrate that AS605240 may represent a promising novel agent for the treatment of inflammatory bowel disease by suppressing leukocyte infiltration as well as by immunoregulating the imbalance between proinflammatory and anti-inflammatory cytokines.

The aim of this study was to investigate the relationship between anti-fibrotic effect of Panax notoginseng saponins (PNS) and serum cytokines in rat hepatic fibrosis. Hepatic fibrosis induced by carbon tetrachloride (CCl(4)) was studied in animal models using SD rats. Liver index, serum alanine amino transferase (ALT), aspartate amino transferase (AST), transforming growth factor-beta1 (TGF-beta1), tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and interleukin-10 (IL-10) were measured, respectively. Liver index and the degree of liver fibrosis were also determined. Our results showed that the levels of ALT, AST and liver index in PNS-treated group were markedly lower than those in model group. PNS therapy also significantly attenuated the degree of hepatic fibrosis, collagen area and collagen area percent in liver tissue. Furthermore, the levels of serum TGF-beta1, TNF-alpha and IL-6 were strikingly reduced in PNS-treated group compared with model group while the production of IL-10 was up-regulated. These findings demonstrate that PNS has certain therapeutic effects on hepatic fibrosis probably by immunoregulating the imbalance between pro-fibrotic and anti-fibrotic cytokines.

To examine the relationship between effect of vascular endothelial growth factor (VEGF) on epithelial-myofibroblast transition (EMT) of HK-2 cells and changes in expressions of bone morphogenetic protein-7 (BMP-7) and inhibitor of DNA binding/differentiation (Id) 2, Id3.The cultured HK-2 cells were co-treated with transforming growth factor-beta1 (TGF-beta1) (5 ng/ml) and VEGF165 (0.1, 1, 10, 100 ng/ml), or with TGF-beta1 (5 ng/ml) and VEGF receptor-1 neutralized antibody (10 microg/ ml), and were also co-treated with TGF-beta1 (5 ng/ml) and VEGF165 (100 ng/ml) with or without activin receptor-like kinase 6 (Alk6)/Fc Chimera (2 microg/ml, to neutralize endogenous BMP-7) for 48 hours. mRNA and protein expressions of alpha-smooth muscle actin (alpha-SMA), E-cadherin, BMP-7, Id2 and Id3 of HK-2 cells were assessed with double-stain immunocytochemistry, real-time PCR and Western blot respectively.Compared with normal controls, alpha-SMA expression significantly increased, while E-cadherin, BMP-7, Id2, and Id3 mRNA and protein expressions markedly decreased in HK-2 cells treated with TGF-beta1 (5 ng/ml) (P0.05). VEGF165 interrupted TGF-beta1 induced alpha-SMA expression in a dose-dependent manner and upregulated BMP-7, Id2 mRNA and protein expressions of the cells (P0.05). alpha-SMA expression increased, while E-cadherin, BMP-7, and Id2 expressions decreased further in HK-2 cells co-treated with TGF-beta1 and VEGFR1 antibody compared with normal controls (P0.05). When endogenous BMP-7 was neutralized with Alk6/Fc Chimera in the cells co-treated with TGF-beta1 and VEGF165, alpha-SMA expression upregulated (P0.05), while Id2 was not changed.VEGF165 may partially inhibit TGF-beta1-induced EMT of HK-2 cells in vitro. This effect is related to the upregulated expressions of BMP-7 and Id2. Id2 may be upregulated directly by VEGF165, but not related to BMP-7.

The aim of this study is to examine the effect of mycophenolate mofetil (MMF) on epithelial-myofibroblast translation (EMT) in adenine-induced chronic renal failure (CRF) rat model and the role of vascular endothelial growth factor(VEGF) and inhibitor of differentiation (Id2 and Id3) in EMT in the rat kidney.Sixty-four male Wistar rats were randomly assigned to the following groups: normal control (n = 16), CRF (n = 24) and MMF(n = 24). CRF was induced by gastric gavage of adenine (125 mg kg(-1) d(-1)) to rats for eight weeks. CRF rats were treated with MMF (15 mg kg(-1) d(-1)) as "MMF" group. The rats were sacrificed at week 2, 4, 6 and 8, respectively. Urinary protein and serum creatinine levels were measured, and the histopathologic degrees of interstitial fibrosis were evaluated in Masson-stained sections. Expressions of alpha-smooth muscle actin (alpha-SMA), transforming growth factor beta1 (TGFbeta1), VEGF and Id (Id2 and Id3) in the kidney tissue were assessed by immunohistochemistry, RT-PCR and/or Western blot methods.The urinary protein level in MMF group was evidently lower than that in CRF group (P0.01), whereas no statistically significant difference was observed in serum creatinine level between the two groups. Renal interstitial fibrosis was reduced significantly with MMF treatment (P0.01). Expression of alpha-SMA in MMF group was lower than that in CRF rats at week 6, 8 (P0.01), while expression of TGFbeta1 was decreased markedly at week 2, 4, 6 (P0.01). The expressions of VEGF in MMF rats were increased significantly at week 6, 8 (P0.01), and Id2, Id3 in MMF rats were increased significantly at week 4, 6 (P0.05).MMF may ameliorate chronic renal fibrosis and EMT in adenine-induced CRF rats. This effect of MMF on EMT is probably related to upregulation of VEGF, Id2 and Id3 expressions and suppressing overexpression of TGFbeta1 in renal tissue. The exact mechanism needs to be studied further.

To observe the effects of Yiqi Kaimi Recipe (YQKMR), a traditional Chinese compound medicine, on gastrointestinal motility and neuropeptides in rats with colonic slow transit constipation.Eighty specific pathogen free rats were included, and there were sixty rats with slow transit constipation (STC) and twenty normal rats. Sixty rats with STC were randomly divided into untreated group, cisapride-treated group and YQKMR-treated group, and twenty normal rats were allocated to normal control group. Percentage of carbon propelling, contraction frequency and amplitude of the muscle segments in vitro, and expressions of nitric oxide synthase-1 (NOS1) and substance P proteins were observed.Compared with the normal control group, the percentage of carbon propelling, and the contraction frequency and amplitude of the muscle segments in the untreated group decreased obviously (P0.05) The percentage of carbon propelling, and the contraction frequency and amplitude of the muscle segments in the YQKMR-treated group and cisapride-treated group were higher than those in the untreated group (P0.05). The expression of NOS1 protein in the untreated group was higher than that in the normal control group, YQKMR-treated group and cisapride-treated group. However, there were no significant differences between the YQKMR-treated group and the normal control group (P0.05). The expression of substance P protein in the untreated group was lower than that in the normal control group, and there was no significant difference between the YQKMR-treated group and the normal control group too (P0.05).Yiqi Kaimi Recipe could promote colon motility by increasing the contraction frequency and amplitude of the smooth muscle, and this may be due to its effect of regulating the expressions of NOS1 and substance P proteins in the enteric plexus.

Objective To evaluate the efficacy and safety of thread-dragging through fistula method in treating patients with simple anorectal fistula. Methods In this multi-centered, prospective, and randomized controlled clinical trial, 244 patients with simple low or high anorectal fistula were randomly divided into study group (with the method of thread-dragging through fistula) and control group (with the method of incision or thread-drawing). The healing time and curative rate of anorectal fistula, and the integral calculus of clinical symptom and life quality evaluations before and after treatment were all examined. The maximal anal canal squeeze pressure was measured to compare the therapeutic safety between these two groups. The health economical benefits were also assessed to determine which therapeutic method was more economical. Results The curative rate of simple low and high anorectal fistula were of no significant differences between the study group and the control group. The healing time of simple low anorectal fistula in the study group and the control group were (22.26+/-8.67) d and (31.41+/-11.39) d respectively, while the healing time of simple high anorectal fistula in the study group and the control group were (24.73+/-8.15) d and (32.20+/-12.60) d respectively, and there revealed significant differences between these two groups. Each integral calculus of clinical symptom evaluation in the study group was not obviously different from those in the control group besides the integral calculus of anal sphincter function. The integral calculus of life quality between the study group and the control group of simple low anorectal fistula had no significant differences. The integral calculus of anal sphincter function and confidence in treatment in the study group of high anorectal fistula were better than those in the control group. The hospitalization expense of the study group was remarkably lower than that of the control group. The maximal anal canal squeeze pressure in the study group after treatment was not reduced obviously as compared with that in the same group before treatment, while it was decreased significantly in the control group after treatment as compared with those in the same group before treatment and in the study group after treatment. Conclusion The method of thread-dragging through fistula in treating simple low and high anorectal fistula can shorten the course of the disease, save the hospitalization expenses, improve the life quality of the patients, and protect the anal sphincter function.

OBJECTIVE To study the effects of Wenshen Jianpi Recipe (WSJPR, a traditional Chinese medicine for warming kidney and invigorating spleen) on chronic wound healing and the mechanism. METHODS Ninety-six SD rats were randomly divided into 4 groups, with 24 rats in each group, and back wound was made in the rats. For rats in 3 of the 4 groups, hydrocortisone injection was administered to induce chronic wound. Rats in 2 of the 3 groups were treated with WSJPR and Xinpukang Granules (XPKG) respectively, and the rats in the other group were untreated. The rats in the fourth group were taken as control. The wound healing time and the width of new epidermis were observed, and the histomorphological changes and cell cycle of the granulation tissue, and the protein expressions of epidermal growth factor (EGF), transforming growth factor-beta(1) (TGF-beta(1)) and fibronectin (FN) in the granulation tissue were tested with immunohistochemical technique and flow cytometry. RESULTS The wound healing time of the WSJPR-treated and XPKG-treated groups was (17.0+/-1.9) and (18.8+/-1.9) d respectively, much shorter than that of the untreated and control groups (P

HLCDG1, which locates in chromosome 5q33 (between D5S436 and D5S470),is a novel gene that our laboratory has cloned recently. The expression of HLCDG1 gene was significantly down-regulated or deleted in the primary lung carcinoma. This study was designed to observe if HLCDG1 has the potential to suppress growth of lung carcinoma cells.The recombinant plasmid, pcDNA3.1(+)/HLCDG1, was constructed and subsequently transfected into A549 cells through liposome transfection. The A549 cells stably expressing HLCDG1 gene were established by G418 selection. RT-PCR was used to demonstrate the expression of HLCDG1 gene. Furthermore, the cell proliferation assay, the soft agar assay, and the tumorigenesis assay were used to analyze the malignant phenotype of the HLCDG1-transfected cells.The HLCDG1-transfected cells exhibited the expression of HLCDG1 mRNA by RT-PCR. The population double time (PDT) of HLCDG1-transfected group, vector-transfected group, and nontransfected group were 70.0 hours, 43.3 hours, and 39.5 hours, respectively; the difference between HLCDG1-transfected group and the other two groups was significant (P0.05). The colony formation rates of HLCDG1-transfected group, the vector-transfected group, and nontransfected group were 8.5%, 29.0%, and 35.0%, respectively. The rate of HLCDG1-transfected cells was markedly lower than those of the other two groups (P0.05). Moreover, these clones were injected into athymic nude mice. After 43 days, they were killed, and their tumors were isolated. These tumors weighed 0.120g, 0.612g, and 0.924g, respectively.The expression of HLCDG1 in A549 cells may have the potential to suppress tumor cell growth and the tumorigenesis of A549 cells transplanted in nude mice. These results suggested that HLCDG1 gene might be a good candidate of tumor suppressor gene correlated with lung carcinoma.

High frequent loss of hetero- zygosity (LOH) of 3p, 5q, 6q, 9p, 10q, 11p, 13q, 17p, and 19p in lung carcinoma was detected by comparative genomic hybridization (CGH) and genomic-wide scan for analysis of genetic alteration with microsatellite allelotying. It was indicated that there might be some other unknown tumor susceptible genes or suppressor genes likely to be involved in lung carcinoma development and progression. The aim of this study was to clone the full-length cDNA of LXDD1,an expressed sequence tag(EST) isolated by mRNA differential display, which is significantly down-regulated in lung carcinoma and represents a novel gene.Differential expression of LXDD1 in lung carcinoma was confirmed by Northern blot analysis, the expression of the LXDD1 in human normal tissues and the size of the transcription of the LXDD1-representative gene were also determined using MTN (Multiple Tissues Northern Blots). The putative full-length cDNA of the EST-representative gene was cloned and analyzed by bioinformatics. In addition, differential reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of the novel gene in various cancer cell lines, primary lung carcinomas, and matched normal lung tissues.The full length cDNA with no homology to any reported genes in the database of GenBank was successfully cloned and named HLCDG1 (Human lung carcinoma deleted gene 1, GenBank accession number AF447582). A transmembrane protein with 166 amino acids was deduced to come from the open reading frame of the 3113 bp full-length cDNA, HLCDG1 gene was confirmed to be located at chromosome band 5q33 by alignment of electric polymerase chain reaction (e-PCR).HLCDG1 is a novel gene down-regulated in lung carcinoma, which may be involved in the development of lung carcinoma.

It was reported that a locus of high frequency loss of heterozygosity was found at 3p14-25 of nasopharyngeal carcinoma (NPC) cells. This study was designed to identify the expressed sequence tags (ESTs) associated with NPC at chromosome 3p21 in order to clone new candidate NPC-associated genes at the locus.The expression of relative ESTs at 3p21 was determined in nasopharyngeal carcinoma tissues and normal nasopharyngeal epithelia using ESTs homology analysis with bioinformatics in computer network combined with reverse transcription-polymerase chain reaction (RT-PCR). The expression of EST was detected in other normal tissues and tumor cell lines using Northern blot analysis.Comparing with expression of normal nasopharyngeal epithelia, EST (N31985) was down-regulated in 60.00% (3/5) nasopharyngeal carcinoma cell lines and 47.06% (16/34) nasopharyngeal carcinoma tissues (P0.05).N31985 is down-regulated in NPC, suggesting it may play a role in NPC carcinogenesis.

The molecular mechanism of the pathogenesis of lung carcinoma is unclear; because key genes related to lung carcinoma have not been identified. This study was designed to isolate and identify differentially expressed cDNA sequences in human lung carcinoma for cloning lung carcinoma-related genes.Using mRNA differential display method combined with cloning and reserve Northern dot blot and sequencing and RT-PCR, differentially expressed cDNA sequences were isolated between two lung cancer cell lines, two adult lung carcinoma tissue and paired normal tissue of trachea epithelia.Sixteen differentially expressed cDNA fragments were isolated, subsequent cloning of sixteen differentially expressed cDNA fragments were confirmed by reverse Northern dot blot and sequencing and BLAST analysis. Two of these were shown to be novel gene sequences that had not been reported. Eight of the remaining cDNA sequences homologize to known genes; additionally two differentially expressed cDNA sequences were confirmed by RT-PCR.Using mRNA differentially display, The authors had successfully isolated ten differentially expressed cDNA sequences from human lung carcinoma, these cDNA sequences might be involved in the pathogenesis of lung carcinoma.

The isolation of the genes related to laryngeal carcinoma(LC) is necessary for revealing mechanisms of carcinogenesis and genetic predisposition to LC. The mRNA differential display method was used to compare and analyze mRNAs prepared from two adult laryngeal carcinoma tissues and paired tumor-adjacent normal tissues. A total of twenty-two differential display experiments was performed and thirty-five cDNA fragments differentially expressed in normal or malignant laryngeal epithelial tissues were identified. Differential expression of six of these thirty-five cDNA fragmens was confirmed by reverse northern dot blot. Subsequent cloning of six differentially expressed cDNA fragments and sequencing and BLASTn analysis resulted in the identification of twelve distinct cDNA sequences. Four of these were shown to be novel gene sequences that have not been reported. Eight of the remaining cDNA sequences showed sequence homology to those previously reported. The differential expression of these twelve cDNA sequences in the carcinoma or normal tissue of the larynx were confirmed by fixing the twelve cDNA sequences on the membrane, followed by the hybridization with the total cDNA probes from laryngeal carcinoma or normal tumor-adjacent tissues and by differential RT-PCR. These results suggest that these cDNA sequences might be involved in carcinogenesis of laryngeal carcinoma.