1.Change of tear film function in seasonal allergic conjunctivitis between acute exacerbation and non-onset phase
Xuan LI ; Zhixin JIANG ; Peng HAO
Chinese Journal of Postgraduates of Medicine 2013;36(24):15-17
Objective To investigate the change of tear film function in patients with seasonal allergic conjunctivitis (SAC) between acute exacerbation and non-onset phase.Methods This was a prospective controlled study.Functional assessment of tear film was performed in 30 eyes of 15 patients with SAC (SAC group) between acute exacerbation and non-onset phase and 15 healthy controls (control group).The tear film function included tear film break-up-time (BUT),Schirmer I test (SIt) and tear film interferometer imager measurement.Results BUT was significantly decreased in SAC group on acuteexacerbation compared with that on non-onset phase and control group [(6.97 ± 1.56) s vs.(11.27 ± 1.39),(12.00 ± 1.11) s],and there was significant difference (U =20.50,P =0.000;U =1.00,P =0.000).Moreover,BUT in SAC group on non-onset phase was similar as control group(U =322.00,P > 0.05).There was no significant difference in SIt among SAC group on acute exacerbation and non-onset phase and control group(P > 0.05).In tear film interferometer imager measurement,80.0%(24/30) was Ⅰ-Ⅱ grade,20.0%(6/30) was Ⅲ grade in control group,20.0%(6/30) was Ⅰ-Ⅱ grade,80.0%(24/30) was Ⅲ-Ⅴ grade in SAC group on acute exacerbation,60.0%(18/30) was I-Ⅱ grade,40.0%(12/30) was Ⅲ-Ⅴ grade in SAC group on non-onset phase,and there was significant difference between SAC group on acute exacerbaiion and SAC group on non-onset phase,control group (x2 =19.27,P =0.000; x2 =8.40,P =0.004),and there was no significant difference between SAC group on non-onset phase and control group (x2 =1.98,P>0.05).Conclusion SAC can cause the instability of tear film during the acute exacerbation,whereas this instability can be recovered within the non-onset phase of S A C,which is close to the normal control
2.Protective effects of thymosin β4 on ethanol-induced rabbit corneal epithelial cell damage in vitro
Zhixin, JIANG ; Peng, HAO ; Xin, TANG ; Xuan, LI
Chinese Journal of Experimental Ophthalmology 2016;34(2):108-114
Background Laser assisted subepithelial keratomileusis (LASEK) is one of surgical procedures for refractive correction.Dilute alcohol that is used for the removal of epithelium during LASEK induces the apoptosis of corneal epithelial cells.Researches showed that thymosin β4 (Tβ4) can arrest apoptosis, but whether Tβ4 plays inhibitory effect on ethanol-induced damage of corneal epithelial cells is still unelucidated.Objective The aim of this study was to investigate the protective effects of Tβ4 on ethanol-induced rabbit corneal epithelial cell damage in vitro.Methods The corneal tissue of deendothelium was obtained from 10 New Zealand white rabbits.Corneal epithelial cells were cultured in vitro by using explant culture method.Cultured cells were identified by detecting the expression of keratin 12 and connexin 43 with reverse transcription PCR (RT-PCR).The cells of second generation were collected and divided into 4 groups.The cells were regularly cultured in the normal control group, and Tβ4 was added in the culture medium at the final concentration of 1 μg/ml in the Tβ4 treated group.Ethanol-induced rabbit corneal epithelial cell damage models were established by adding PBS containing 20% alcohol in medium for 20 seconds in the model group,and Tβ4 was added in the medium of model cells at the final concentration of 1 μg/ml in the model+Tβ4 group.The survival rate of the cells was detected by MTT assay, and the apoptosis rate of the cells was examined by TUNEL method.The relative expression levels of cyclin D1 mRNA and cyclin-depensent protein kanase 4 (CDK4) mRNA in the cells were detected by real-time flurescenee quantitative PCR.The content of bcl-2 protein in the cells was detected by ELISA assay.Spectrophotometry was employed to measure the activity of caspase-3.The study complied with the Regulation for the Adminstration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The mean cell survival rate was (52.1 ± 14.07) % in the model group,which was significantly reduced in comparison with 100% of the normal control group and (77.7± 19.60) % of the model+Tβ4 group (P=0.001 ,P=0.035).TUNEL staining revealed more positive cells in the model group and the model+Tβ4 group,and the percentage of TUNEL positive cells was (30.0±6.7)% in the model+Tβ4 group, showing an evident decline in comparison with (42.4±4.0)% of the model group (P=0.01).The relative expression levels of cyclin D1 mRNA and CDK4 mRNA were 0.93±0.17 and 0.88±0.09 in the model+Tβ4 group, which were significantly higher than 0.68±0.05 and 0.54±0.07 in the model group (P=0.027,0.002).ELISA assay exhibited that bcl-2 content in the cells was considerably lower,and caspase-3 activity was significantly higher in the model group than those in the model+Tβ4 group (P =0.030,0.021).Conclusions Tβ4 plays a protective effect on rabbit corneal epithelial cells from apoptosis by lowing the caspase 3 activity and increasing bcl-2 content in ethanoldamaged rabbit corneal epithelial cells.In addition, Tβ4 promotes the regrowth of corneal epithelial cells by upregulating the expression of cyclin D1 and CDK4.
3.Recognition of Specific Antigens by Specific IgG and IgE During Anaphylactic Shock Induced by Echinococcus granulosus in Sheep
Hong ZHENG ; Zhixin XU ; Gexiong YANG ; Hao WEN
Chinese Journal of Parasitology and Parasitic Diseases 1997;0(06):-
Objective To determine the specific recognition of Echinococcus granulosus (E.g.) cyst fluid crude antigen (EgCF) and antigen B (EgB) by serum specific IgG and IgE using Western blotting during anaphylactic shock induced by E. g. in sheep, and to investigate the importance of defined characteristics and molecular weight of the specific antigens. Methods EgCF was obtained through local slaughterhouses in Urumqi from the cysts of infected sheep liver. Western blotting was used to analyze total specific IgG and IgE antibodies in serum from sheep infected with E.g. using either crude antigen of E. g. and EgB, and to understand specific recognition of hydatid cyst antigens by serum total IgG and specific IgE. Results SDS-PAGE and Western blotting showed that there were differences between EgB and EgCF in electrophoresis pattern. EgB was recognized by IgG from sera of infected sheep in a series of bands with molecular weight ranging from 31, 43 to 66.2 kDa. No binding of IgG against EgCF was observed in any serum from the infected sheep during anaphylactic shock. In contrast, specific IgE antibodies in E. g. infected sheep obviously recognized the single 43 kDa subunit of EgCF, but no binding of specific IgE against EgB was observed in sera of the infected sheep. Conclusion EgCF is consisted of antigenic components in which there is a specific antigen against IgE with a molecular weight of over 43 kDa. This component may lead to an anaphylactic shock induced by E. g. . EgB is a specific antigen against the total IgG but not to the specific IgE.
4.Study on the Level of Specific IgG, IgG1 and IgE During Anaphylactic Shock in Sheep Induced by Echinococcus granulosus
Hong ZHENG ; Zhixin XU ; Gexiong YANG ; Hao WEN
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(01):-
Objective To investigate the change of specific antibodies IgG, IgG1 subclass and IgE in sheep infected with Echinococcus granulosus(E.g) during anaphylactic shock, and to observe antigen B reactivity against IgG antibody in E.g\|infected sheep. \ Methods\ Antigen B and crude antigen were prepared with E.g cyst fluid (EgCF) from infected sheep. The enzyme\|linked immunosorbent assay(ELISA) was used for detecting the level of specific IgG, IgG1 and IgE during anaphylactic shock in sheep induced by E.g. \ Results \ The level of specific IgG, IgG1 and IgE was significantly higher in the infected sheep after 6 months than that of the uninfected control group (P
5.Synthesis and in vitro bioIogicaI activity of 3-phenyI-3-pyrroIyIpentane deriva-tives
Meng XU ; Zhixin GE ; Meixi HAO ; Can ZHANG
Journal of China Pharmaceutical University 2016;(1):30-37
A new series of 3-phenyl-3-pyrrolylpentane derivatives are synthesized through modifying the structure of lead compound LG19055,a nonsecosteroidal vitamin D receptor(VDR)agonist.The VDR-agonistic ability of target compounds was measured indirectly by evaluating the differentiation ability of HL-60 cell.The results showed that compounds 13a,13c,13d,13h,13i,13j have excellent VDR-agonistic ability(EC50 <50 μmol /L), especially for compound 13j (EC50 =0.10 μmol /L),which was more potential than that of lead compound LG190155.Their proliferation inhibitory activities in vitro were evaluated by MTT assay in MCF-7,PC-3,Caco-2, HepG2 and L02 cell lines.Compound 13a exhibited significant inhibitory effects on HepG2 cell line(IC50 =0.11μmol /L).Moreover,the inhibitory effect of compound 13a on non-tumor liver L02 cell line was relatively weak (IC50 =15.24 μmol /L ),suggesting that compound 13a has selective inhibitory effects on liver cancer cells.Additionally,HL-60 cell differentiation-inducing activity and the inhibitory effect of cancer cells were posi-tively related.
6.The protective effects of ethanol extract of Trillium tschonoskii Maxim. on hypoxia-ischemia brain damage in neonatal rats
Yong QIU ; Renpeng LI ; Su LIU ; Zhixin TAN ; Longquan CHEN ; Hong LIU ; Hao WU
Chinese Pharmacological Bulletin 2016;32(11):1548-1552
Aim To investigate the effect of Trillium tschonoskii Maxim ( TTM ) ethanol extract on hypoxia ischemia brain damage ( HIBD ) in neonatal rats and potential mechanisms. Methods Fifty healthy SD rats of 7 day-old were randomly divided into three groups:the sham operation group ( n=10 ) , the model group ( n=20 ) and TTM treatment group ( n=20 ) , which received 3-day intraperitoneal injection of normal saline or ethanol extract of TTM respectively. TTC staining and Nissl staining were performed to detect the cerebral ischemia area and neuronal death. Western blot was used to detect the expression of Bcl-2 and Bax. Re-sults The brain tissue of model group was slightly swollen, and white necrotic zone induced by ischemia occured on the right side of the brain, while the brain morphology of TTM treatment group was good. After TTC staining, ischemia zone was clearly seen on the right side of the brain in model group, while after TTM treatment, the size of ischemic zone was decreased. Compared with the model group , Nissl staining showed the neuronal cells increased in TTM treatment group. Western blot showed the expression of Bcl-2 protein in TTM group increased than that in HIBD model group ( P <0. 01 ) , while the expression of Bax protein de-creased ( P <0. 01 ) . Conclusion TTM therapy is beneficial for HIBD,which may be related to reducing neuronal apoptosis.
7.Influence of Bupleurum Liver-Coursing Powderon behavior,neuronal apoptosis and autophage in hippocampus of depressive rats
Aijun XU ; Hao LIU ; Yanxia TIAN ; Yufang ZHAO ; Quan KAN ; Zhixin CHEN ; Haitao WANG
Journal of Jilin University(Medicine Edition) 2014;(4):801-804
Objective To observe the effect of Bupleurum Liver-Coursing Powder on the neuronal apoptosis and autophage in hippocampus of depression model rats, and to explore the mechanism of its anti-depression effect. Methods 60 male SD rats were randomly divided into control group,model group and drug intervention group (n=20).The depression model was produced by giving the rats chronic unpredicted mild stress.The rats in model group and control group received the same volume of normal saline,and the rats in drug intervention group received Bupleurum Liver-Coursing Powder solution on the basis of model.The depressional behavior was examined using sucrose preference test,tail-suspension test and Morris water maze.Flow cytometry was employed for the detection and quantification of the apoptotic cells in the hippcampus. The expressions of LC-3 and Beclin 1 were detected using Western blotting method. Results The pencertages of sucrose preference of the rats in model group and drug intervention group were significantly lower, while the tail-suspension immobility time and the excape latency time were higher than those in control group (P<0.05);the apoptotic rates of cells,the ratios of LC3-Ⅱ/LC3-Ⅰand the expression levels of Beclin-1 were higher than those in control group (P<0.05).Compared with model group,the percentage of sucrose preference of the rats in drug intervention group was increased (P<0.05),the tail-suspension immobility time and the excape latency time were decreased (P<0.05),and the apoptotic rate of cells,the ratio of LC3-Ⅱ/LC3-Ⅰand the expression level were also decreased (P<0.05).Conclusion Bupleurum Liver-Coursing Powder has significant anti-depression effect, which may be related to inhibiting the apoptosis and degrading the autophage of neuros.
8.Endoplasmic reticulum stress signaling pathway and its association with liver diseases
Yao LI ; Zhixin WANG ; Hao WEN
Journal of Clinical Hepatology 2020;36(2):464-467
Endoplasmic reticulum stress (ERS) manifests as the aggregation of misfolded and unfolded proteins in the endoplasmic reticulum lumen and disorder of calcium balance and can activate the signaling pathways involved in unfolded protein response, endoplasmic reticulum overload reaction, and sterol regulatory cascade response. ERS can not only exert a protective effect by inducing the expression of endoplasmic reticulum molecular chaperones such as glucose-regulated protein 78 and glucose-regulated protein 94, but also induce cell apoptosis. At present, there is still no systematic understanding of ERS involvement in the development and progression of liver diseases. This article summarizes the research advances in ERS-related signaling pathways and related liver diseases and elaborates on the role of ERS-mediated cell apoptosis in liver diseases. The intervention of ERS signaling pathways may provide a reference for the research and treatment of liver diseases in the future.
9.Influence of endoplasmic reticulum stress on the secretion of some inflammatory mediators in hepatic alveolar echinococcosis
Yao LI ; Jingchao WANG ; Jiaqi YUAN ; Hao WEN ; Ying ZHOU ; Haining FAN ; Zhixin WANG
Journal of Clinical Hepatology 2021;37(1):131-134
ObjectiveTo investigate the influence of endoplasmic reticulum stress (ERS) on some inflammatory mediators during the progression of hepatic alveolar echinococcosis (HAE) and its clinical significance. MethodsA total of 15 patients with HAE who underwent partial liver resection in Qinghai University Affiliated Hospital from June 2018 to September 2019 were enrolled, and the marginal zone of HAE lesion was resected as AE group; 15 normal liver tissue samples collected during the same period of time were selected as control group. Western blot and qRT-PCR were used to measure the protein and mRNA expression of protein kinase R-like ER kinase (PERK), CCAAT/enhancer-binding protein homologous protein (CHOP), caspase-12, and glucose-regulated protein-78 (GRP-78), and q-PCR was used to measure the mRNA expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor (TNF). The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups, and the t-test was used for comparison of normally distributed continuous data between two groups; a Pearson correlation analysis was performed to investigate the correlation between two variables. ResultsCompared with the control group, the AE group had significantly higher protein expression levels of PERK, CHOP, caspase-12, and GRP78 (U=4.165, 3.461, 2.577, and 3.344, all P<0.001) and their mRNA expression levels (t= 34003, 4.461, 53.573, and 55.224, all P<0.001). The AE group had significantly higher mRNA expression levels of IL-1β, IL-6, and TNF than the control group (t=6.090, 12.578, and 53.573, all P<0.001). The protein expression levels of PERK, CHOP, caspase-12, and GRP-78 were positively correlated with the mRNA expression levels of IL-1β, IL-6, and TNF (all r>0.700, all P≤0.05). ConclusionPositive correlation is observed between the activation of ERS and inflammatory mediators in HAE, and excessive activation of ERS can change the secretion of several inflammatory mediators to exacerbate liver injury, while further studies are needed to clarify the specific mechanism.
10.Correlation between A11084G in mitochondrial ND4 gene and Parkinson's disease in female patients from northern China
Qin XU ; Xiaoguang LUO ; Qiaohong CHU ; Xiaoni ZHAN ; Zhixin HUO ; Hao PANG
Chinese Journal of Neuromedicine 2017;16(4):392-397
Objective To explore the potential relationship between 6 single nucleotide polymorphisms (SNPs) of mitochondrial DNA (mtDNA) coding region and Parkinson's disease (PD) in northern Chinese population.Methods Samples from 353 patients with PD and 389 control participants,collected in our hospital from April 2010 to November 2015,were genotyped by multiplex polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) techniques.Six SNPs (T4216C within the ND1 gene,A4917G within the ND2 gene,A11084G within the ND4 gene,A12308G within the tRNA gene,A13966G within the ND5 gene,and G15928A within the tRNA gene) in mtDNA were amplified in multiplex PCR systems and subsequently genotyped by digestion with endonucleases.Results An efficient,rapid,and economical way of multiplex genotype SNPs was successfully established.No significant differences were observed in the 6 loci and corresponding haplotypes between PD and control cohorts (P>0.05).After stratification by gender,however,frequencies of allele 11084G and the haplotype T-A-G-A-A-G in female group with PD were all higher than those in control ones,with significant differences (P<0.05).Conclusions The 6 SNPs may not be susceptible factors in the northern Chinese population.The A11084G missense mutation might be related to PD in female patients.