Objective:To make the isolation and 2-DE analysis of mitochondria,in order to prepare the following proteomic study on mitochondria of hepatoma-cell. Methods:Mitochondria was separated from cell homogenate by means of density gradient centrifugation,and 2-DE was conducted to examine the protein profile of mitochondrial preparations.Results:The increase in purity of mitochondria was found to be 12 times by density gradient centrifugation.Mitochondrial proteins were displayed well in the 2-DE pattern after purification.Conclusion:The isolation procedure is practicable,which provides a basis for the following proteomic study on mitochondria.

0.05).The positive expression rate of CXCR4 was 96% in cancer cells in regional metastasized lymph nodes.And its expression was lower than that in primary tumors(P

0.05);The levels of basal FINS showed a significant differences between group A and group B(P0.05).Conclusions Intensive insulin treatment to newly diagnosed type 2 diabetes patients can control blood glucose rapidly to a desired level,and can recover the function of B cell better than traditional oral hypoglycemic agents by dissolving the virulence of glucose rapidly.

Objective: To study the effect of Resen heche tablets (RSHC) on tonyfing and warming Shen Yang. Methods: The mice model of Shen Yang deficiency was established by injecting hydrocortisone. The body weight, autonomic activity and swimming time in ice water in model mice were observed. The level of superoxide dismutase (SOD) in red cell and lipid peroxide (LPO) in plasma of aged rats were analyzed. The effect on anti stress in mice was examined. Results: RSHC had the remarkable effect to improve body weight and autonomic activity and to prolong swimming time in ice water in model mice, and to increase SOD activity and reduce LPO in blood of aged rats, and to elevate the ability of anti fatigue and anoxic tolerance in mice. Conclusion: RSHC drug has the effect on tonifying and warming Shen Yang; can promote growth, enhance organism activity, elevate the adaptation to circumstances and protect injury from free radicals.

Objective:To study the therapeutic effect and mechanism of autologous platelet-rich plasma (PRP) on rabbit traumatic optic neuropathies (TON) and retina.Methods:Forty adult New Zealand white rabbits were selected to establish the optic nerve clamp injury model in their right eyes.According to the random number table method, 36 New Zealand white rabbits with effective model were randomly divided into model control group, normal saline control group and PRP group, 12 for each group.Another 12 healthy rabbits served as the normal control group.Rabbit autologous blood was collected to prepare PRP.The retrobulbar 20 μl PRP/20 μl saline solution injection was administered every two days near the injury after modeling according to grouping.The injection was carried out for 10 times.There was no other interference administrated to the model control group except the normal anti-infective treatment.No interference was given to the normal control group.At 30 and 60 days after modeling, the eyeballs and optic nerves of right eyes were harvested through sacrificing the animals by anesthetic overdose, three eyes for each time.Histopathological assessments were performed to observe the morphological changes of retina and optic nerve, and to evaluate the changes of retinal ganglion cells (RGCs) density and retinal nerve fiber layer (RNFL) thickness.Immunohistochemistry was used to assess the expressions of apoptosis factors caspase-3 and B cell lymphoma-2(Bcl-2). Quantitative real-time PCR and Western blot were used to detect the mRNA and protein expressions of brain-derived neurotrophic factor (BDNF) and growth associated protein-43 (Gap-43). This study protocol was approved by the Experimental Animal Ethics Committee of Wuhan University (No.E2019072805). The use and care of animals complied with ARVO statement.Results:The thickness of RNFL and number of RGCs at 30 days and 60 days after modeling were (6.60±1.16) μm, (6.89±1.21) μm, (13.00±1.00)/field of vision, (20.00±2.65)/field of vision in the PRP group, respectively, and were (4.80±0.43)μm, (2.18±0.23)μm, (6.33±0.58)/field of vision, (10.33±1.53)/field of vision in the model control group, respectively.The number of RGCs in the PRP group at 60 days was higher than that at 30 days after modeling, the number of RGCs in the PRP group was higher than that in the model control group, the thickness of RNFL in the PRP group was higher than that in the model control group; and the differences were statistically significant (all at P<0.05). At 30 and 60 days after modeling, the positive expression A value of caspase-3 protein in the normal saline group and model control group were higher than those in the normal control group and PRP group, while the positive expression A value of Bcl-2 protein in the PRP group was higher than those in the model control group and normal saline group, and the differences were statistically significant (all at P<0.05). The mRNA level and protein content of BDNF and Gap-43 in the retina and optic nerves at 30 days and 60 days after modeling in the PRP group were higher than those in the model control group, and the differences were statistically significant (all at P<0.05), but the mRNA and protein expression levels of BDNF and Gap-43 in different tissues in the PRP group at 60 days after modeling were lower than those at 30 days after modeling ( P<0.05). Conclusions:PRP can effectively inhibit the apoptosis of RGCs and the secondary injury of the retina after optic nerve injury, promote cell anti-apoptosis effect of RGCs, thereby retard the damage of the retina and optic nerve after TON, and also promote the repair of optic nerve and retina through upregulating the expression of nerve growth factors.

Coronary heart disease has become a major threat of human health, and its incidence rate remains surprisingly high.In recent years, technology on its diagnosis and treatment has developed greatly.The present article made a review on related new diagnosis and treatment so as to provide reference for clinical diagnosis and treatment of coronary heart disease.

Oligodeoxynucleotide containing unmethylated cytosine phosphate-guanosine motif(CpG ODN) may induce high expression of CD80, CD86, CD83, HLA I and HLAⅡ molecules on dendritic cells(DC) and stimulate DC to produce high level of IL-6, IL-12, TNF-? and IFN-?. CpG ODN is demonstrated in vivo to be a very potent adjuvant for Th1 cells, regulating Th0 cells to develop toward Th1 cells. Its role for DC is characteristics of CpG ODN sequence specificity and species specificity. CpG ODN is, at present, considered as a pathogen associated molecular pattern which binds its specific receptor,Toll-like receptor 9,then functions through TLR/IL-1R signaling pathway. It may represent a new therapeutic drug for broad applications in infectious disease, autoimmune disease, allergy and cancer therapy.

The expression of c-met stimulated by high glucose in human renal tubular epithelial cells and the role of HGF/c-met system in diabetic nephropathy were examined. The proximal tubular epithelial cells were cultured in vitro under different conditions. MTT was used for the detection of cellular proliferation and RT-PCR was employed for measurement of c-met mRNA level. Our results showed that under different conditions, there were no significant differences in the proliferation of proximal tubular epithelial cells 12 h and 24 h after the culuture (P>0.05). The proliferation of proximal tubular epithelial cells showed a significant change 96 h after the culture and the cellular proliferation induced by hepatocyte growth factor (HGF) was very active (P<0.05). Moreover, no significant difference in the expression of c-met mRNA was found 12 h after the culture under different conditions (P>0.05), while 24 and 96 h after the culture, a persistent and significantly higher expression of c-met mRNA was found in HGF-induced proliferation. It is concluded that addition of exogenous HGF could inhibit the apoptosis induced by high-level glucose, promote the proliferation of proximal tubular epithelial cells, and induce the expression of c-met. Our study suggests that local up-regulation of HGF/c-met system plays an important role in the repair of renal damage in diabetic nephropathy.

Objective To investigate the effects of adiponectin on cell activity and expression of T-cadherin in human retinal pigment epithelial(hRPE) cells cultured with high glucose.Methods ①The hRPE cells were divided into three groups:normal glucose group(5.5mmol/L glucose),high glucose group (33mmol/L glucose),high glucose plus adiponectin group (33mmol/L glucose+2.5ug/ml adiponectin).48h later,the cell activity was observed by MTT assay.②After the above groups were cultured respectively for 24h,48h,72h again,the expression of T-cadherin mRNA was observed by RT-PCR. Results ①Cell activity in high glucose group significantly decreased. Cell activity adiponectin group was higher than that in the high glucose group.②With the extension of time ,the expression of T-cadherin decreased in high glucose group,but increased in adiponectin group. Conclusion Adiponectin may be protect the hRPE cells by T-cadherin.

Objective To explore effect on the proliferation and apoptosis of Hela cells in vitro by using vascular endothelial growth factor-C(VEGF-C) antisense oligonucleotides (ASODN). Methods VEGF-C ASODN was transfected into Hela cells by liposome-mediated; the cells transfected with the oligodeoxynuclecotide (SODN) and saline were used as control groups. The efficiency of transfection was detected by fluorescence microscope. The inhibitive rate of cell growth was detected with MTT methods. Apoptosis and cell cycle were evaluated using FCM. The mRNA expression of VEGF-C was determined by RT-PCR, and the expression of VEGF-C was determined by western blotting. Results VEGF-C ASODN had been transfected into Hela cells by liposome-mediated. The index of apoptosis after transfection 48 h was (19.39±1.81)%. G2/M phase cell increased after transfection, meanwhile the expression of VEGF-C degraded on the level of mRNA and protein (P<0.05). Conclusion Transfeeted with VEGF-C ASODN could down-regulate the expression of VEGF on the level of mRNA and protein, block the cell circle, inhibit cell proliferation and induce apoptosis in Hela cells in vitro.