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

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 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 examine the expression of protein kinase C (PKC), connexin43(Cx43) and non-phosphorylated Cx43 in ovarian cancer, and discuss the role of phosphorylated of Cx43 in chemoresistance in ovarian cancer. Methods We examined the expression of Cx43, non-phosphorylated Cx43 and PKC in ovarian cancer tissue by immunohistochemistry, and compared their expression in chemosensitivity group and ehemoresistance group. Cisplatin resistant ovarian cancer cell line SKOV3/DDP cells were treated by staurosporine (a kind of PKC inhibitors). Then expression of Cx43, non-phosphorylated Cx43 and PKC were tested. Meanwhile, we tested chemosensitivity of SKOV3/DDP cells by ATP bioluminescence tumor chemosensifivity assay (ATP-TCA). Results (1) Immunohistochemically,the rates of positive expression of Cx43 and non-phospharylated Cx43 were 54%, 14% respectively in the chemoresistance group, which were 83%, 59% in the chemosensitivity group respectively (P<0.05). The rate of positive expression of PKC in 28 chemoresistance ovarian cancer cases (64%) was higher than that in 29 chemosensitivity cases (31% ,P<0.05). Both of them were significantly lower in ehemoresistanee group than in chemosensitivity group (P<0.05). In addition, the expression of PKC was negatively correlated with the expression of Cx43 and non-phosphorylated Cx43. The correlation coefficients were -0. 626 and -0. 714, respectively (P<0.05). (2) Immunohistochemically, PKC was down regulated, and Cx43 and non-phosphorylated Cx43 were up regulated in SKOV3/DDP cells after staurosporine treatment. The longer the staurosporine worked, the more expression of Cx43 was. (3) By ATP-TCA, SKOV3/DDP cells were resistant to paclitaxel and cisplatin. The tumor growth inhibition was higher in the group of paclitaxel or cisplatin combined staurosporine than in the group of paclitaxel or cisplatin alone. The sensitivity was intermediate in the group combined with low concentration staurosporine (1×10-8 moL/L), and the sensitivity was high in the group combined with high concentration staurosporine (1×10-7 mol/L). Conclusions Phosphorylation of Cx43 caused by PKC leads to decrease in the expression of Cx43. This effect makes ovarian cancer cells less chemosensitive. Phosphorylation of Cx43 caused by PKC can be inhibited by staurosporine.

The pathological changes in myocardial cell of diabetic rats were consistent with diabetic cardiomyopathy. The levels of adiponectin and the expression of T-cadherin in diabetic group were increased gradually,and higher than in normal control group

Objective To investigate the association of expressions of neutrophil extracellular trap (NET) and B lymphocyte in renal tissue with lupus activity in patients with lupus nephritis (LN). Methods Immunohistochemistry was.used to detect the expression of NET (citrullinated histone H3 as the marker) and the infiltration of B lymphocyte (CD19 as the marker)in renal specimens of three groups [LN group,n=20; minimal change disease (MCD) group,n=8;healthy control group,n=3].The chronic index and SLE-disease activity index (SLE-DAI) in renal tissues of LN and their correlations with NET and B lymphocytes were examined. Results The expression of NET was not found in the renal tissues of healthy control group and MCD group,while it increased significantly in LN group,especially in glomeruli with moderate and severe mesangial cells proliferation, cellular crescents, and tubulointerstitium with inflammatory cell infiltration.Compared with other types of glomeruli,the expression of NET was significantly upregulated in glomeruli with moderate and severe mesangial cell proliferation (P＜0.01).In the glomeruli with moderate and severe glomerular mesangial cell proliferation,the mean number of =NET positive cells was positively correlated with renal pathological active index (r=0.620,P=0.004),the score of SLE-DAI (r=0.492,P=0.027) and the mean number of NET positive tubular cells (r=0.558,P=0.011).In renal interstitium,the NET positive cells were positively correlated with CD19 positive B lymphocytes (r=0.573,P=0.008) and renal pathological chronic index (r=0.645,P=0.002). Conclusion NET is widely expressed in the renal tissues of lupus nephritis,which may play a role in the active damage of glomeruli.

Objective To investigate the expression of activator protein-1 (AP-1) in PBMCs from patients with dermatomyositis (DM) and glucocorticoid effect on the expression.Methods PBMCs were isolated from 15 normal human controls and 20 patients (including 12 patients with primary DM who had never received glucocorticoid treatment and 8 patients with recurrent DM who had stopped glucocorticoid treatment for more than 1 month),and classified into two parts:one was cultured in RPMI 1640 medium with dexamethasone of 80 μmol/L and 10％ calf serum for 48 hours,and the other was frozen at -80 ℃ and reserved.Electrophoretic mobility shift assay (EMSA) was carried out to detect the activity of AP-1 in these cells.Results The expression (grayscale area) of AP-1 in the PBMCs from the normal controls was 4.93 ± 0.15 mm2.A significant decrease was induced by the treatment with dexamethasone in the expression (grayscale area) of AP-1 from PBMCs of patients with primary DM and those with recurrent DM (5.59 ± 0.39 vs.30.23 ± 0.49 mm2,5.69 ± 0.39 vs.34.79 ± 0.61 mm2,F =13812.64,P ＜ 0.01 ).The activity of AP-1 was statistically higher in the PBMCs from patients with recurrent DM than in those from patients with primary DM before the treatment with dexamethasone (P ＜ 0.01 ).Conclusions The enhanced activity of AP-1 may be an important factor inducing the inflammatory reaction in and recurrence of DM.Glucocorticoids may suppress the AP-1 activity to a certain degree.

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.

Before irreversible neuronal damage occurs, quickly and securely recanalize vessels and restore blood flow to save the ischemic penumbra tissue are the main therapeutic target for acute ischemic stroke (AIS). Currently, the method for recanalization in early AIS is intravenous thrombolysis, how ever, the effect of intravenous thrombolysis is often not satisfactory because of the reasons, such as its short treatment time w indow , low vascular recanalization rate, and high incidence of bleeding complications. In recent years, w ith the research and development of mechanical thrombectomy device and the development of interventional technology, the therapeutic time w indow of AIS has been significantly prolonged, the vascular recanalization rate has increased significantly, the clinical outcome has improved significantly, and the treatment of mechanical thrombectomy has show n good application prospects.