2.Protective effect of carnosine on cognitive impairment induced by subcortical ischemic vascular dementia
Jing MA ; Jian ZHANG ; Zhong CHEN
Chinese Pharmacological Bulletin 2014;(4):496-500,501
Aim Subcortical ischemic vascular demen-tia ( SIVD ) induced by chronic hypoperfusion due to small-artery disease is a common cause of vascular de-mentia ( VaD) , which is recognized as the second most prevalent type of dementia. The aim of this study was to determine whether carnosine played a protective role in cognitive impairment induced by permanent occlu-sion of the right unilateral common carotid arteries ( rUCCAO ) in SIVD. Methods Adult male mice ( C57BL/6 strain ) were subjected to rUCCAO, and treated with carnosine or saline. Locomotor test, open field test, hot plate test, freezing test and Morris water maze were performed after rUCCAO. Results There were no differences among rUCCAO group, carnosine group and sham group for total distance traveled in lo-comotor test. In the open field test, carnosine (200, 500 mg · kg-1 ) significantly revised the decrease of latency spent in the center induced by SIVD . There were no differences between rUCCAO and sham groups for the pain threshold. In freezing test, rUCCAO in-duced a significant reduction in content memory, which was completely reversed by treatment of carnosine. In Morris water maze training trials, rUCCAO-treated mice showed prolonged escape latency in acquisition phase, carnosine ( 200, 500 mg · kg-1 ) markedly shortened the escape latency. Conclusion These data suggest that carnosine has a neuroprotective effect on cognitive impairment induced by rUCCAO in mice.
3.Metformin induced apoptosis in breast cancer cell by decreasing the expression of Foxp3
Wenjun HUANG ; Jian CHEN ; Jing TIAN
Chongqing Medicine 2014;(28):3749-3752
Objective To research the effects of metformin on proliferation and apoptosis in human breast cancer MCF-7 cells and investigate the role of Foxp3 in the effect.Methods The inhibit rate of MCF-7 cells was measured by MTT assay after 48 h metformin treatment(5 mmol/L、10 mmol/L、20 mmol/L).Metformin-induced cell apoptosis was measured by Hoechst 33258 stai-ning and flow cytometry.The expression levels of Foxp3 and caspase-3 mRNA were detected by real-time PCR.Expression of Foxp3 protein was detected by Western blot.Results Metformin markedly inhibited proliferation of MCF-7(P<0.05)compared with control group.And the early apoptotic rates increased to 3.76%,8.96%,and 18.67% with 5 mmol/L、10 mmol/L、20 mmol/L metformin treatment,respectively.Metformin increased caspase-3 mRNA expression levels,and decreased the expression levels of Foxp3 mRNA and protein expression levels(P<0.05,vs.control).Conclusion Metformin could inhibit the proliferation and in-duce apoptosis of MCF-7,and its mechanism maybe related to down-regulated expression of Foxp3.
4.The clinical analysis of ultrasound biomicroscope scanning photography in anterior segment structure changes in blunt eye injury
Jian ZENG ; Jing CHEN ; Lingzhu FANG
Chinese Journal of Ultrasonography 1993;0(02):-
Objective To clinical analyze the changes of ocular anterior segment structure using ultrasound biomicroscope(UBM) and their clinical values in blunt eye injury.Methods Sixty-nine eyes with no-opening wound in blunt eye injury were examined by UBM(Paradigm Model P40).Scanning photography of central and 3,6,9,12 clock hour were recorded and printed,other clock hours were scanned if needed.These records were studied with clinical findings.Results UNM findings showed that 27 eyes had hyphema,22 eyes had anterior chamber angle recession,16 eyes had iridodialysis,19 eyes had local luxation of lens,5 eyes had vitreous henia in anterior chamber,19 eyes had cyclodialysis,23 eyes anterior choroidal detachment and 9 eyes had anterior vitreous haemorrhage.Of the 69 eyes,62 eyes had combined structure changes showed above,which account(89.9)%.Conclusions UBM is a practical tool for diagnosis of anterior segment structure changes in blunt eye injury,especially when there are corea opacity,hyphema,cyclodialysis,ocular hypotension and others.It can provide exact informations for the diagnosis and treatment,which can not be showed by routine ocular examination and B-scanning ultrasonography.
5.Study on gene expression of PTH/PTHrP receptor in patients with renal impairment
Jian ZHANG ; Jing CHEN ; Weiyu ZHU
Chinese Journal of Nephrology 1997;0(06):-
Objective To investigate the changes of parathyroid hormone receptor in patients with different stages of renal disease and its possible underlying mechanisms. Methods Relatively quantitative RT/PCR method was used to detect PTH receptor mRNA expression in renal specimens obtained through biopsy or operation. Results The levels of PTH receptor mRNA were significantly decreased in all patients with chronic glomerulonephritisx moderate renal failure severe renal failure and acute renal failure, which was correlated with the extent of renal impairment. Conclusion The downregulation of renal PTH receptor occurs much earlier than the changes of renal function., plasma PTH., serum phosphate and calcium in the course of human renal disease.
6.Recombination expression and immunoprotective effects of the P6 outer membrane protein of nontypeable Haemophilus influenzae
Xiangpeng CHEN ; Jian WANG ; Lili LI ; Jing CHEN ; Zhenlong ZHANG
Chinese Journal of Microbiology and Immunology 2010;30(4):349-354
Objective To clone and express recombinant outer membrane protein P6, determine its optimal expression conditions and to investigate the immunoprotective effects of the P6 protein on mice. Methods The P6 gene of nontypeable Haemophilus influertzae(NTHi) was amplified by PCR from the NTHi genome and cloned into expression vector pET-32a (+) to generate the pET-32a-P6 recombinants. They were confirmed by nuclease digestion and sequence analysis. The verified recombinant was transformed into E. coli BL21 (DE3). Its optimal expression conditions were determined such as engineering strains, the concentration of IPTG, inducing temperature, inducing time, different medium etc. The recombinant protein was purified by Q Sepharose~(TM) XL ion exchange and gel filtration chromatography. The protein was analyzed by SDS-PAGE, Western blot and sequencing. BALB/c mice were immunized with recombinant protein be-fore challenged by NTHi through intraperitoneal injection. Then the mortality rate of different group was com-pared. Results The recombinant P6 of NTHi was successfully constructed and expressed in E. coli at a rel-atively high level. The purity was up to 95% after purification. The relative molecular mass of the protein is 14 145. 848. The recombinant protein was confirmed to show specific reaction on the antiserum through Western blot. The animal experiments showed the mortality rates of immunization groups were significantly lower than that of the control group (P < 0.05). Conclusion The successful expression of the recombinant P6 will be very helpful for the further study on development of vaccine, its purified, immunological activity and antibody preparation.
7.The role of B7-H1mRNA in immune privilege of homogeneity
Xiaohuan HAUNG ; Jing WU ; Jian YU ; Linjiang CHEN ; Ming MA
Chinese Journal of Organ Transplantation 2012;33(9):567-570
Objective To clarify the role of B7-H1 in the immune privilege after corneal transplantation in homogeneity variant mice.Methods We established the experimental animal model of allograft mice by using C57BL/6 mouse as donor and Balb/c mouse as recipient.We allocaated the mice with long time survival (>50 days) corneal graft into survival group,mice with rejection occurring in 50 days into rejection group,and normal C57BL/6 mice into control group.The transplanted corneal grafts were obtained for future reference at the 8th week after transplantation in survival group,and the time of rejection in rejection group.The expression of B7-H1 mRNA was detected by using immunohistochemistry and real time quantitative PCR (RT-PCR),and the relationship between B7-H1 and the immune privilege after corneal transplantation was analyzed. Results The B7 H1 mRNA was highly expressed in epithelium and endothelium of corneal grafts both in survival and control group,in comparison to an obviously lower expression in rejection group.The relative expression level of B7-H1 mRNA was 200.0 ± 11.5 in survival group,44.7 ± 10.8 in control group,and 6.9 ± 12.0 in rejection group,respectively. There were statistically significant differences among the three groups (F=241.164,P<0.01 ).The was a significant correlation between the level of B7-H1 mRNA and occurrence rate of rejection in corneal graft (P<0.01 ).Conclusion The results suggest that the immune privilege after corneal transplantation might be mediated by B7-H1,which plays an important role in maintaining the state of corneal immune privilege.
8.Influences of antenatal administration of taurine on cerebral neurogenesis of fetal rats with intrauterine growth restriction
Chen HUI ; Li JIAN ; Liu JING ; Liu LI ; Liu NA
Chinese Journal of Perinatal Medicine 2011;14(10):612-617
Objective To provide the experimental evidence for clinical application of taurine,rat model of intrauterine growth restriction (FGR) was made to investigate influence of prenatal administration of taurine on neurogenesis.Methods Fifteen pregnant rats were divided into control,FGR model and taurine groups,5 rats for each group.Rats in the control group were supplied with unlimited food and drink while the other two groups were fed by 40% food intake of the control group throughout pregnancy.Since gestational day 12,taurine (100 mg/kg) was added into diet of taurine group every day until term delivery.Brain tissues were obtained immediately after baby rats were born.Expression of proliferating cell nuclear antigen (PCNA),neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) of brain tissue was measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry; meanwhile,numbers of PCNA-immunoreactive cells in subventricular zone,subgranular zone and cerebral cortex were compared with ANOVA test or q test.Results Levels of PCNA mRNA and GFAP mRNA expression in FGR group were significantly higher than those of control group (PCNA mRNA:1.002±0.011 vs 0.894 ± 0.040,P<0.01; GFAP-mRNA:1.012±0.013 vs 0.913±0.012,P<0.01).Compared to FGR model group,mRNA expressions of PCNA and GFAP in taurine groups were higher (1.090±0.029,P<0.01 ; 1.034±0.011,P>0.05).There was a significant decrease in the expression of NSE mRNA in FGR group compared with control group (0.796±0.036 vs 1.582±0.057,P<0.01),while the expression in taurine group (0.933±0.027) was significantly higher than that in FGR model group (P < 0.01).PCNA immunoreactive cells were mostly distributed in subventricular zone,followed by subgranular zone and cerebral cortex.Conclusions Prenatal application of taurine could enhance neurogenesis of FGR newborn rats and improve survival of neurons to ameliorate FGR brain damage.
9.Construction and identification of specific shRNA interference plasmid vector targeted to uPAR gene
Jing WANG ; Jian CHEN ; Ming MA ; Yuan ZHAO ; Jie ZHANG
China Oncology 2009;19(12):904-909
Background and purpose: Urokinase-type plasminogen activator receptor is related to invasion and metastasis of tumor. Inhibition of uPAR expression in tumor cells results in reducing its metastasis. This study was aimed to construct an expression vector with short hairpin RNA (shRNA) of uPAR, which could pave the way for RNAi-mediated tongue squamous cell carcinoma therapy. Methods: Genome sequences of uPAR gene were retrieved from Genhank and cDNA was designed to code expression of shRNA for uPAR gene. The cDNA was synthesized and inserted into the eukaryotic expression vector pWH1, and the recombinant pWH1-uPAR expression vector was identified by enzyme cutting method. Then, pWH1-uPAR expression vector was transfected into tongue squamous cell carcinoma Ts cells by Lipofectomine 2000. At last, the expression of uPAR in Ts cells transfected with pWH1-uPAR expression vector was observed by RT-PCR, immunocytochemistry staining and Western blot. MTT assay was performed to measure the proliferation of Ts cell. Results: The uPAR shRNA eukaryotic expression vector was successfully constructed. Compared with Ts cells and Ts cells transfected with plasmid pWH1, the Ts cells transfected with pWHI-uPAR expression vector showed a lower mRNA and protein expression of uPAR. The inhibition rate of proliferation was 32.9% of Ts cells by transfected with pWHl- uPAR. Conclusion: The constructed uPAR shR.NA expression vector could inhibit the expression of uPAR in tongue squamous cell carcinoma, which may be helpful for further research on the function of uPAR and provide effective methods for therapy of tongue squamous cell carcinoma.
10.DNA damage of bone marrow cells in mice induced by radon exposure
Jing SUN ; Bingyan LI ; Yuejin CHEN ; Jihua NIE ; Jian TONG
Chinese Journal of Radiological Medicine and Protection 2008;28(5):449-451
Objective To study the DNA damage of bone marrow cells of mice after exposure to Radon.Methods Twenty-four mice were randomly divided into four groups, one control group and three experimental groups with the cumulative doses of radon at 27 WLM (low dose group), 52 WLM (middle dose group) and 105 WLM ( high dose group). DNA damage induced by radon in bone marrow of mice was detected by methods of single cell gel electrophoresis (SCGE), micronucleus(MN) and laser scanning confocal microscope (LSCM) observation. Results The DNA strand breakage, rate of MN and apoptosis increased significantly in the high dose group, but not in the middle and low dose groups. Conclusions Exposure to radon could induce DNA damage in bone marrow cells of mice at high levels.