1.Detection and analysis of the Rhodopsin gene in a consanguineous Chinese Han autosomal dominant retinitis pigmentosa family
Xue-juan, CHEN ; Xiang, GAO ; Chen, ZHAO ; Kan-xing, ZHAO
Chinese Journal of Experimental Ophthalmology 2013;(6):602-607
Background Rhodopsin (RHO) gene is the most common disease gene for autosomal dominant retinitis pigmentosa (adRP),one of the main pathogenesis is that misfolded mutant RHO proteins accumulate in the endoplasmic reticulum and cause endoplasmic reticulum stress (ERS).Objective This study aimed to determine the genetic basis for a consanguineous Chinese Han adRP family.Methods This study procedure complied with Helsinki Declaration.All participants in the family were investigated under the informed consent.Regular ocular examination was performed on the patients in this family.Next-generation sequencing (NGS) was carried out to screen the mutations in 189 genes associated with hereditary retinal diseases (HRDs).After being analyzed and filtered,variations detected by NGS were validated by Sanger sequencing and evaluating of pathogenicity.The wild-type RHOWT and mutant RHOP53Rwere cloned into the vector pEGFP-N1.Then the two plasmids were transfected into adult retinal pigmentosa epithelium cell line(ARPE19) and human embryo kidney 293 line (HEK293) to observe the location of rhodopsin-GFP fusion protein in cells,and the expression of ERS related protein XBP1 in the cells was detected by quantitative-PCR and Western blot.Results This family included 5 generations with the typical adRP characteristics.Genetic analysis identified a heterozygous variation,p.P53R in RHO gene,which was fully cosegregated in the family.Wild-type RHOWT-GFP fusion proteins showed the green fluorescence on the endoplasmic reticulum and cytomembrane,but the misfolded mutant RHO-GFP fusion protein gathered only in endoplasmic reticulum.Compared to wild-type RHOWT,the XBP1 was activated and increased by (1.28 ±0.09) fold.The introns of 26 bases in XBP1 mRNA were removed in the HEK293 cells with mutant RHO-GFP fusion protein,and the expression of XBP1 was stronger in the HEK293 cells with mutant RHO-GFP than that in HEK293 cells with wild type RHO-GFP and cells with blank pEGFP-N1 plasmid.Conclusions Heterozygous variant RHO p.P53R is very likely the pathogenical mutation in the adRP family.The RHOP53R mutant rhodopsin protein can not be delivery effectively from the endoplasmic reticulum to the cell membrane,and these proteins accumulate in the endoplasmic reticulum,which causes ERS.
2.Research advances in molecular epidemiology and vaccines of Coxsackievirus A16.
Xiang-Peng CHEN ; Xiao-Juan TAN ; Wen-Bo XU
Chinese Journal of Virology 2014;30(4):483-488
Epidemics of hand, foot and mouth disease (HFMD) have mainly been caused by Coxsackievirus A16 (CVA16) and Enterovirus A 71 (EV-A71), which circulated alternatively or together in the affected area. CVA16 has caused numerous outbreaks and epidemics in multiple countries and geographical regions, and has become an important public health problem. Based on an analysis of the complete VP1 coding region, all CVA16 strains can be divided into genotypes A, B1, and B2. Furthermore, genotype B1 can be divided into subgenotypes B1a, B1b, and B1c. After 2000, no reports of genotype B2 virus strains have been reported. All of the CVA16 strains reported in mainland China have belonged to subgenotypes B1a and B1b. Most CVA16-associated infections cause only mild symptoms; however, some CVA16 infections can lead to severe complications and even death. Vaccination is considered to be the most effective method to control the transmission and infection rate of this virus. A number of research groups are studying various vaccine types, including inactivated vaccines, genetic engineering vaccines, and DNA vaccines, amongst others. In this review, an overview is provided of the research advances in molecular epidemiology and vaccines of CVA16.
Animals
;
China
;
Coxsackievirus Infections
;
epidemiology
;
immunology
;
prevention & control
;
virology
;
Enterovirus A, Human
;
classification
;
genetics
;
isolation & purification
;
Humans
;
Molecular Epidemiology
;
Viral Vaccines
;
administration & dosage
;
genetics
;
immunology
3.The effect of high-fat diet and catch-up growth on gastric emptying and plasma glucagon-like peptide-1 concentration in rats
Lulu CHEN ; Weihong YANG ; Juan ZHENG ; Ling YUE ; Xiang HU
Chinese Journal of Internal Medicine 2009;48(8):655-659
a greater impact,which could account for some pathophysiological changes caused by catch-up growth.
4.Purification and Biological Characteristics of Bacteriocin-like Substance Produced by Lactobacillus buchneri KLDS1.0364
Han-Sheng GONG ; Xiang-Chen MENG ; Hong-Juan LIU ;
Microbiology 1992;0(02):-
The purpose of this study is to obtain effective and stable bacteriocin-like substance from lactic acid bacteria. Lactobacillus buchneri KLDS1.0364,which was isolated from fermented cream,a traditional dairy product in Inner Mongolia,could produce bacteriocin-like substance. The bacteriocin-like substance produced by KLDS1.0364 was partially purified and the characteristics were studied. The bacteriocin-like substance was purified by SP-Sepharose fast flow cation exchange chromatography. The molecular weight of the bacteriocin-like substance was 21.6kD,as determined by tricine-SDS-PAGE. The bacteriocin-like substance remained stable after 30 min at 121℃ and after 2 h of incubation during pH 2~10. Complete inactivation of antimicrobial activity was observed after treatment of the bacteriocin-like substance with several proteinases. Treatment with catalase and ?-amylase did not result in any changes of antimicrobial activity. The mode of action of the bacteriocin-like substance is bactericidal. It exhibited a broad spectrum of antagonistic activity against various species of Gram-positive bacteria,Gram-negative bacteria and fungi.
5.Effects of behavioral inhibition system/behavioral activation system and emotion regulations on depression of adolescents
Yanzhang LI ; Yun XU ; Zi CHEN ; Juan XIANG
Chinese Journal of Behavioral Medicine and Brain Science 2013;22(10):926-928
Objective To explore the effects of behavioral inhibition system/activation system (BIS/BAS) and emotion regulations on depression of adolescents.Methods 800 adolescents from an university and a middle school were chosen by cluster random sampling,and were surveyed by BIS/BAS scale,Beck depression inventory(BDI),center for epidemiologic studies depression scale (CESD) and cognitive emotion regulation questionnaire (CERQ).The data were analyzed by pearson correlation,F test and multiple stepwise regression analysis.Results (1) BDI and CESD were correlated positively with BIS (r =0.39,0.41,P < 0.01),self-blame,rumination,catastrophizing,blame others and perspective(r =0.19-0.50,P < 0.05-0.01),and negatively with planning,reappraisal (r =-0.08-0.24,P<0.05).(2)The scores of severe depression group were highest(15.45 ±2.57,P<0.01) in BIS,lowest in BAS-drive(9.60 ±2.99,P<0.01),and nonadaptive emotion regulations were used mostly,and adaptive emotion regulations were done leastly.(3) Catastrophizing,BIS,reappraisal,rumination,refocusing and self-blame explained 35% of total variance,and catastrophizing and BIS explained the largest proportion,respectively 25% and 4%.Conclusion BIS/BAS and emotion regulations may be important factors influencing non-clinical depression of adolescents.
6.Inhibition of the recombinant human endostatin adenavirus on experimental choroidal neovascularization in rats
Li-Juan, CHEN ; He-Xiang, SONG ; Lin, MIAO
International Eye Science 2017;17(6):1037-1041
AIM: To investigate the inhibition of the recombinant human endostatin adenavirus (Ad-Es) on the experimental choroidal neovascularization(CNV) models by intravitreous injection.METHODS: Experimental CNV models were induced by semiconductor laser in 30 male Brown Norway(BN) rats and randomly divided into 3 groups with 10 rats in each group.At 21d after photocoagulation, the single administration group were given intravitreous injection with Ad-Es 0.01mL;the repeated administration group were given intravitreous injection with Ad-Es 0.01mL and a repeated injection 7d later;the saline control group were given intravitreous injection with saline 0.01mL.At 7d after final administration, the leakage of fundus fluorescein angiography (FFA) was observed.Various CNV areas were measured by using laser confocal microscopy of choroidal flatmount method.Pathology and ultrastructure were observed with light microscopy, the expressions of CD105 were measured by immunohistochemistry.RESULTS: The leakage of CNV of the administration group abviously decreased as compared with those in the saline group, the leakage of repeated administration group decreased compared with that of single administration group (P<0.05).Laser confocal microscope quantitative CNV analysis showed that CNV area of the administration group was significantly smaller than that of control group, the area of repeated administration group was smaller than that of single administration group (P<0.05).Under the light microscope, the vascular endothelial cell number and quantity of the administration group were significantly lower than that of the control group, the cell number of repeated administration group was lower than that of single administration group.CD105 expression of the administration group was significantly weaker than that in the saline group;the expression of repeated administration group was weaker than that of single administration group.CONCLUSION: Ad-Es can effectively inhibit semiconductor laser induced CNV in BN rats, and the inhibition effect of repeated administration group is better than that of single administration group.It may be a useful new method in the treatment of CNV.
7.Effects of infrasound on the auditory function of guinea pigs
Juan JIN ; Jing LIU ; Xiang MU ; Jing-Zao CHEN ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(05):-
Objective To observe the effects of exposure to infrasound on auditory function of guinea pigs as reflected by the brain stem response (ABR) and distortion product otoaeoustic emission(DPOAE).Methods Seventy-two Guinea pigs were used in this study,of them,12 served as controls and 60 were divided into 2 experi- mental subgroups.ABR and DPOAE were detected after exposure to infrasound stimulation at 16 Hz,90 dB or 16 Hz,130 dB for 2 hours a day for 1 ,7,14,21 and 28 days.Results The threshold of ABR after exposed to infrasound at 16 Hz,90 dB in 1 day and 28 days was higher than the controls (P0.05),but significant difference was observed with 16 Hz 130 dB infrasound exposure on the 21st and,28th days when compared with the controls (P
8.Effects of viaminate on the proliferation and differentiation of a human keratinocyte cell line HaCaT
Xiang CHEN ; Ting-Ting LUO ; Juan SU ; Bei-Bei ZHANG ;
Chinese Journal of Dermatology 2003;0(11):-
Objective To investigate the effects ofviaminate on the proliferation and differentiation of HaCaT cells,a human keratinocyte cell line.Methods Cultured HaCaT cells were treated with various concentrations (2,5,10,15,20,25 and 30?g/mL) of viaminate for various durations.The cell proliferation was assessed by MTT method,the changes of cell cycle and apoptosis rate by flow cytometry,the changes of keratin 10 and involucrin mRNA expressions by semi-quantitative reverse transcription PCR.Results The proliferation of HaCaT cells was inhibited by the treatment with viaminate of≥2?g/mL for 48 h,and the inhibition rate was raised with the increase of treatment time and dosage.The viaminate of 30?g/mL inhibited the proliferation of HaCaT cells by 57.67% and 82.00% at 48 and 72 h after the incubation respectively,and elevated the mRNA expression of involucrin from 40.80% to 156.12%,decreased the mRNA expression of keratin 10 from 96.46% to 14.60%.The mRNA expression of involucrin increased with the elevation of viarninate dosage.Under the treatment with viaminate for 48 h,the cell population at G_1 phase significantly increased,that at S and G_2 phases decreased;the switching of G_1 to G_2 was inhibited;but the cell apoptosis was not affected.Conclusion Viaminate could inhibit the proliferation and induce the differentiation of keratinocytes.
9.Studies on the Central Regulatory Mechanism of Acupuncture-moxibustion Treatment for Gastric Mucosal Injury
Juan XIANG ; Guo CHEN ; Fei LI ; Liting XIANG ; Ying CHEN ; Jun YU ; Zhou YANG ; Liang PENG
Shanghai Journal of Acupuncture and Moxibustion 2016;35(12):1484-1487
This article provides the possible mechanism of acupuncture-moxibustion repair of gastric mucosal injury from central regulation and puts it forward that the nucleus of solitary tract is the primary regulation center for the injury repair and has the effect of collecting and integrating information. In addition, it is put forward that neural regulation is a main regulatory mechanism for the injury repair and besides, endocrine, immune and humoral regulations participates in the modulation and gastric mucosal repair involves a composite regulatory mechanism in which the central nervous system, neuroendocrine-immune network and neurohumoral regulation take part.
10.Expression of CD147,cyclophilin A and cyclophilin B in psoriatic lesions
Xiang CHEN ; Juan LI ; Juan SU ; Yi-Xi WU ; Jing CHANG ; Sheng PENG ; Ye-Hong KUANG
Chinese Journal of Dermatology 2003;0(10):-
Objective To study the expression and pathologic significance of CD147,cyclophilin A (CyPA)and cyclophilin B(CyPB)in psoriatic lesions.Methods Immunohistochemical method was ap- plied to detect the expression of CD147,CyPA and CyPB in skin specimens of 15 patients with psoriasis pustulosa(PP),20 patients with progressive psoriasis vulgaris(PPV),and 20 patients with inactive psori- asis vulgaris(IPV).Immunoreactivity intensity distribution index(IRIDI)was calculated to assess the expres- sion intensity of CD147,CyPA and CyPB.Results CD147,CyPA and CyPB were detected in all speci- mens.The IRIDI scores of CD147 and CyPA were significantly higher in keratinocytes of psoriatic lesions than in those of the control specimens(all P0.05).The IRIDI score of CyPB in T lymphocytes of PP lesions was significantly elevated than that in PPV lesions,which was in turn higher than that in IPV lesions(all P0.05).Conclusion CD147,CyPA and CyPB may play a role in the occurrence and development of psoriasis.