Objective To compare the effects of single and complex emotional stimuli on animal model of liver depression syndrome,in order to find out an animal model of liver depression syndrome with more similar clinical manifestations.Methods Based on the previous screening work,different methods,i.e.tail clamping,limb binding,putting a cangue on the neck,were used respectively,or using the three methods in combination to establish rat models of liver depression syndrome.A comparison table of human and rat liver depression syndrome was prepared,a mark sheet of evaluation criteria was made,general condition was observed and body weight was measured,and positive response to drugs and changes of hormonal levels were evaluated to identify whether the models were established successfully.Results Rats in the control group grew well and showed no abnormalities.There were lots of abnormal phenomena in the model group especially in the complex emotional stimulus modeling group.They were maniac with acute stress or depressive with chronic stress,had dry hair,small and less feces,many of the rats had hair loss,red eyes and shortness of breath symptom.They showed a significantly lower body weight gain and higher levels of CRH in hypothalamus and plasma ACTH and CORT.The manifestations were much more severe in the complex-emotional stimuli-induced model group than that in the single-stimulus prepared model groups.The drug-treated group showed considerable alleviation of the symptoms and improvement of the parameters closing to normal levels.Conclutions Establishing the rat model of stagnation of liver depression syndrome by complex emotional stimuli modeling method has good stability,with a short preparation period and high success rate,and presenting a better consistence with clinical liver stagnation of depression syndrome,so as to provide a suitable tool for TCM research.

Background Fungal keratitis has a high incidence in China and its clinical treatment is very difficult,and its etiology diagnosis and appraisal is the premise to improve the prognosis of disease.With the changes of regional environment and climate in recent years,whether the spectrum of fungal keratitis change in South China is remarkable.Objective The purpose of this study was to investigate recent pathogenic distribution of fungal keratitis in South China area.Methods The consecutive fungal culture resuhs of 3 350 purulent keratitis at Zhongshan Ophthalmic Center from January 2009 to December 2014 were retrospectively reviewed.The positive rate of fungal culture,genus or species distribution,seasonal distribution and different term distribution were analyzed.Results The culture-positive rate was 31.34％ in this study (1 050/3 350),and the average culture-positive number was 175 strains per year.In the positive fungus,the highest positive rate was Fusarium SP (32.10％,337/1 050),and followed by Aspergillus SP (25.71％,270/1 050),Heminthosporium SP (14.29 ％,150/1 050) and Mucor SP (9.14％,96/1 050).The fungal culture-positive rate was 36.05％ (367/1 018) in 2009 to 2010,32.45％ (324/1 014) in 2011 to 2012,and 26.86％ (354/1 318) in 2013 to 2014,respectively,with a significant difference among the three periods (x2 =22.37,P＜0.01),showing a decreasing tendency of incidence.Two hundreds and sixtyone strains were isolated from January to March (31.15 ％,261/838),182 strains from April to June (25.53 ％,182/713),237 strains from July to September (30.00％,237/790),370 strains from October to December (36.67％,370/1 009),showing a statistically significant difference among them (x2 =25.19,P ＜ 0.01).The number of infectious strains was most during October to December and fewest during April to June.Conclusions The leading pathogenic fungi of fungal keratitis is Fusarium SP and followed by Aspergillus SP,Helminthosporium SP,Mucor SP in turn.Fungal keratitis is usually prevalent from October to December,and its incidence is still rising in Chinese mainland recently.However,the increasing tendency in South China has been prevented in recent six years.

Objective: To explore the protective roll of ifbroblast growth factor 21 (FGF21) in endoplasmic reticulum stress (ERS) induced rat's H9c2 cardiomyocyte apoptosis with its mechanism. Methods: pcDNA4 was used as gene vector, pcDNA4-FGF21 plasmid was constructed and transfected into rat's H9c2 myocardiocytes for 48 h. ERS model was established by 10 μM tunicamycin (TM) induction for 24 h. The experiment was conducted in 4 groups:①Control group,②TM group, the cells were treated by TM,③pcDNA4-FGF21+TM group,④pcDNA4+TM group. The expressions of FGF21, protein kinase R-like ER kinase (PERK) and c-Jun N-terminal kinases (JNK) mediated apoptosis pathway related protein were measured by Western blot analysis; cell survival rate was examined by CCK-8 method and apoptosis rate was detected by TUNEL technique. Results: pcDNA4-FGF21 vector was successfully constructed and overexpressed in H9c2 myocardiocytes. Compared with Control group, TM group and pcDNA4+TM group had up-regulated endogenous FGF21 expression, increased PERK and JNK mediated apoptosis pathway related protein expression; reduced cell survival rate and elevated apoptosis rate. Compared with TM group and pcDNA4+TM group, pcDNA4-FGF21+TM group had down-regulated PERK and JNK mediated apoptosis pathway related protein expression; increased cell survival rate and decreased apoptosis rate. Conclusion: FGF21 overexpression can reduce ERS induced apoptosis rat's H9c2 myocardiocytes which might be partly related for inhibiting PERK and JNK mediated signal transduction of apoptosis pathway.

Objective To study the genomic molecular organization and genogroup of human metapneumovirus(hMPV) infected infants in Guangzhou of China. Methods Primers were designed on the basis of the genomic sequence of hMPV 00-1 strain(AF371337) in the GenBank, and amplify hMPV genomeby RT-PCR. The PCR-products were cloned to T vector and sequenced, the genomic nucleotide sequences were analyzed with the programs Clustal W/X, DNASTAR and MEGA4. 1. Results The cloned strainhMPVgz01 genome is 13 327 bp in length, the genome contains eight open reading frames in the order 3-N-P-M-F-M2-SH-G-L-5. The genomic sequences of hMPVgz01 strain are compared with those of hMPV in GenBank, revealed that the homology with hMPV group A ranges between 92%-97%, homology with group B is 81%, and with avian metapneumovirus group C is 71%, the highest homology is with BJ1887 strain of genogroup A2b. The N, F, G genes of hMPVgz01 strain are compared with those corresponding genes of hMPV subgroups A1, A2, B1, B2, revealed that the highest homology is also with genogroup A2b. Conclusion The complete nucleotide sequence of hMPVgz01 strain isolated from Guangzhou in China is 13 327 bp in length, GenBank accession No. is GQ153651. Comparison of the genomic sequence and three genes of hMPVgz01 strain with those corresponding sequences of hMPV show the highest homology is with genogroup A2b. Sequence and phylogenetic analysis of the hMPVgz01 strain revegled that this isolate belongs to genogroupA2b.

AIM:To explore the effects of galectin-3 (GAL-3) on the viability, migration and inflammation of human umbilical vein endothelial cells (HUVECs) and the mechanisms.METHODS:The HUVECs were cultured in vitro and treated with GAL-3 recombinant protein at 2 mg/L or GAL-3 short hairpin RNA (shRNA).The HUVECs were divided into normal group, recombinant GAL-3 group, shControl group and GAL-3-shRNA group.The mRNA expression of GAL-3, monocyte chemotactic protein (MCP)-1, IL-6, matrix metalloproteinase (MMP)-9 and cyclin D1 was detected by real-time quantitative PCR,and the protein expression of GAL-3, IL-6 and MCP-1 was detected by Western blot.The secretion levels of MCP-1 and IL-6 in the culture medium were measured by ELISA.The viability and the ability of migration of the HUVECs were examined by CCK-8 assay and wound healing assay.The protein levels of heat shock protein 90 (HSP90), ERK1/2, p-ERK1/2, JNK and p-JNK were determined by Western blot.RESULTS:The expression of GAL-3, MCP-1 and IL-6 at mRNA and protein levels, the mRNA expression of MMP-9 and cyclin D1, and the secretion levels of MCP-1 and IL-6 in the culture medium were significantly higher than those in normal group (P<0.05) after the HUVECs were treated with GAL-3 recombinant protein.However, these molecules mentioned above in GAL-3-shRNA group were significantly lower than those in normal group and negative control group (P<0.05).Compared with normal group, the viability and migration ability of the HUVECs in recombinant GAL-3 group were significantly increased, but the viability and migration ability of the HUVECs in GAL-3-shRNA group were lower than those in normal group and shControl group (P<0.05).In addition, the protein levels of p-ERK1/2 and HSP90 in recombinant GAL-3 group were higher than those in normal group (P<0.05), but those in GAL-3-shRNA group were lower than those in normal group and shControl group (P<0.05).The protein level of p-JNK was not oviously changed among the 4 groups.CONCLUSION:GAL-3 is involved in regulating the cell growth, migration and the release of inflammatory cytokines in vascular endothelial cells, which may be mediated by HSP90-ERK1/2 signaling pathway.

AIM:Chronic exposure to elevated levels of free fatty acids (FFAs) in type 2 diabetes patients is toxic to pancreatic β-cells.Thioredoxin (Trx)-interacting protein (TXNIP), an endogenous Trx-inhibiting protein, is up-regulated by glucose and is a critical mediator of hyperglycemia-induced β-cell apoptosis in diabetes.However, the effects of FFAs on TXNIP are unknown.In this experiment we observed the effect of palmitate on TXNIP expression in cultured INS-1 islet cells and the pathways involved were analyzed meanwhile.METHODS:After the full basis of preliminary experiment of incubating INS-1 cells with palmitate at different concentrations for different time, INS-1 islet cells were cultured with 0.5 mmol/L palmitate for 24 h.TXNIP expression, cell apoptosis, and expression of transcription factors related to TXNIP transcriptional regulation were determined.RESULTS:Compared with control group, the expression of TXNIP at mRNA and protein levels in palmitate group was significantly up-regulated (P<0.01).Cleaved caspase-3/caspase-3 ratio was increased in palmitate group (P<0.05), and the apoptosis of the INS-1 cells was also significantly increased (P<0.01).Palmitate enhanced the phosphorylation of nuclear factor-κB (NF-κB) (P<0.01), and the NF-κB inhibitors, PDTC and SN50, both blocked the palmitate-induced up-regulation of TXNIP expression.CONCLUSION:Saturated fatty acid palmitate enhances the expression of TXNIP.The mechanism of palmitate-induced TXNIP expression may be associa-ted with the increase in NF-κB phosphorylation.

To investigate roles of nitric oxide (NO) signal in accumulations of phenolic acids in abscisic.acid (ABA)-induced Salvia miltiorrhiza hairy roots, S. miltiorrhiza hairy roots were treated with different concentrations of sodium nitroprusside (SNP)-an exogenous NO donor, for 6 days, and contents of phenolic acids in the hairy roots are determined. Then with treatment of ABA and NO scavenger (2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylimidazoline-1- oxyl-3-oxide, c-PTIO) or NO synthase inhibitor (NG-nitro-L-arginine methyl ester, L-NAME), contents of phenolic acids and expression levels of three key genes involved in phenolic acids biosynthesis were detected. Phenolic acids production in S. miltiorrhiza hairy roots was most significantly improved by 100 µmoL/L SNP. Contents of RA and salvianolic acid B increased by 3 and 4 folds. ABA significantly improved transcript levels of PAL (phenylalanine ammonia lyase), TAT (tyrosine aminotransferase) and RAS (rosmarinic acid synthase), and increased phenolic acids accumulations. However, with treatments of ABA+c-PTIO or ABA+L-NAME, accumulations of phenolic acids and expression levels of the three key genes were significantly inhibited. Both NO and ABA can increase accumulations of phenolic acids in S. miltiorrhiza hairy roots. NO signal probably mediates the ABA-induced phenolic acids production.
Abscisic Acid ; pharmacology ; Benzofurans ; metabolism ; Free Radical Scavengers ; pharmacology ; Hydroxybenzoates ; metabolism ; Nitric Oxide ; metabolism ; Phenylalanine Ammonia-Lyase ; metabolism ; Plant Roots ; metabolism ; Salvia miltiorrhiza ; metabolism ; Tyrosine Transaminase ; metabolism

Tanshinone ⅡA is one of the main effective components in Salviae Miltiorrhizae Radix et Rhizoma. It plays a role in the resistance to atherosclerosis by participating in anti-inflammatory in vascular wall, such as the regulating endothelial cell apoptosis and correcting lipid metabolism disorder. This article summarized recent researches of the basic role of tanshinone ⅡA in the resistance to atherosclerosis and provided references for clinical application of antiatherosclerotic effect of tanshinone ⅡA.

Aim To establish the rat model of Spleen-Qi deficiency, analyse the metabolic pathways and investigate the connection between the changed urinary metabolites and Spleen-Qi deficiency, in order to explore the potential mechanisms of Spleen-Qi deficiency.Methods With the binding methods of diarrhea induced by bitter and cold, abnormal of starvation and excessive tiredness, the rat Spleen-Qi deficiency model was established.Then the activity of creatine phosphokinase(CPK) was detected.The endogenous metabolites in the urine were detected by NMR, and the data were analyzed with multivariate and statistical methods.Then the metabolites were selected that could be clearly distinct in the two groups with the fold change value(>1.2) and the P<0.05 of Student′s t-test.Both the pathway analysis and enrichment analysis were performed with Metabo Analyst 3.0.Results Compared with the normal rats, the activity of CPK decreased significantly in model rats(P<0.05).A significant separation appeared in the principal components analysis(PCA) score plot when the control group and the model group were compared, indicating that the Spleen-Qi deficiency model was successfully duplicated.The 33 differential metabolites, which mainly involved in the metabolic pathways, were distinguished from the comparision of Spleen-Qi deficiency model group and control group.The metabolic pathways was related to energy metabolism, amino acid metabolism, nucleotide metabolism and disturbance of gut microbes.Conclusions The main energy metabolic pathways (tricarboxylic acid cycle, glycolysis and liquid oxidation) may be disturbed in Spleen-Qi deficiency rats.The energy supply function is suppressed, which leads to the fatigue and weight loss in rats.

G mutation of SLC26A4, the parents and the second child were carriers of the same mutation, while the fetus had a wild-type form. Conclusion It is feasible to identify deafness related genes by screening for GJB2 and SLC26A4 mutation, thus providing correct prenatal diagnosis and avoiding deaf delivery of baby.