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.

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.

Adult ; Arrhythmogenic Right Ventricular Dysplasia ; physiopathology ; Electrocardiography ; Humans ; Male ; Middle Aged

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.

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.

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.

Objective To investigate the effect of tumor necrosis factor alpha ( TNF-?) in combination with Go6976 on murine liver cancer cells (H22). Method H22 cells were divided into two groups. Each group was further divided into four subgroups. Group 1 was treated with TNF-? 0, 20, 40, 60ng/ml. Group 2 was treated with TNF-? 0, 20, 40, 60 ng/ml and Go6976 (4.6 nmol/ml). The apoptotic rate, protein kinase C alpha (PKC-?) expression and phosphorylation-PKC-? (p-PKC-?) were detected by flow cytometer and Western blotting respectively in 4 h, 8 h and 16 h. Result Treated with TNF-? (0, 20, 40, 60 ng/ml) for 4 h, the apoptotic rates of H22 cells were 2. 44% ? 0. 31 % , 1. 80% ? 0. 32% , 2. 73% ?0. 14% and 3. 05% ?0. 78% respectively, with no change on the expression of PKC-? and p-PKC-?; For 8 h, the expressions of PKC-? and p-PKC-? in H22 cells were up-regulated with increasing concentration of TNF-?; When PKC-? was inhibited with Go6976 at the same time, the apoptotic rates of cells increased significantly, being 2. 90% ?0.39%, 7.76% ?0.35%, 11.43% ?1.05% and 12.96% ?2.44% , respectively. Moreover, PKC-? and p-PKC-? were down-regulated accordingly. When H22 cells were treated with TNF-? only or combined with Go6976 for 16 h, the result was similar to that of 8 h; The apoptotic rate dropped in the group in which PKC-? was inhibited by Go6976 with TNF-? at 60 ng/ml, but the proportion of necrotic cells increased. Conclusion TNF-? up-regulates the expression of PKC-? and p-PKC-? in H22 cells. Inhibiting the activity of PKC-? significantly enhances the toxicity of TNF-? to H22 cells.

AIM: To study the role of scavenger receptor A(SR A) in the uptake of oxidized low density lipoprotein(OxLDL) in mouse peritoneal macrophages(MPM). METHODS: Comparing the difference of the uptake of OxLDL in SR A-deficient and wild-type MPM. RESULTS: The results showed that the binding of OxLDL wasn't apparently reduced in SR A-deficient MPM. The association of OxLDL was reduced by 35.8% and degradation of OxLDL was reduced by 42% in SR A-deficient MPM compared with those in wild-type MPM. CONCLUSION: Studies showed that SR A didn't play an important role in the uptake of OxLDL in MPM. Approximately 70% of the uptake of OxLDL in macrophages is attributable to non-SR A receptor.

Objective To investigate the effects of propofol on the changes in myocardial Toll-like receptor 4 (TLR-4)and TNF-αand NF-κb protein expressions in ischemia-reperfusion injury (I/R).Methods Thirty healthy male SD rats weighing 250-320 g were randomly divided into 3 groups (n=10 for each):Group A,sham operation;Group B,I/R;and Group C,propofol + I/R.In Groups B and C myocardial I/R was induced by occlusion of the left anterior descending artery (LAD)for 30 min,followed by 120 min reperfusion.In Group C propofol was given intravenously 1 0 min before myocardial ischemia,followed by continuous infusion of propofol at 5 mg/(kg·h)until the end of 120 min reperfusion.In Groups A and B normal saline instead of propofol was given. The myocardial tissues were taken at the end of 120 min;ultrastructural changes of myocardial cells were observed under X-ray electron microscope and the expressions of TLR-4 mRNA as well as TNF-αand NF-κb protein were determined.Results Ultrastructural observation under electron microscope showed significantly worsened damage in myocardial tissue structure and mitochondria in Groups B and C compared with Group A.The myocardial expressions of TLR-4 and TNF-αand NF-κb protein were significantly higher in Groups B and C than in control Group A.The myocardial expressions of TLR-4 and TNF-αand NF-κb protein were down-regulated in Group C compared with Group B.Conclusion Intravenous injection of propofol can protect against myocardial damage.Propofol can suppress the increase in myocardial TLR-4 and TNF-αand NF-κb protein expressions induced by I/R.

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.