Objective To investigate the effects of peritoneal injection of cobaltic protoporphyrin Ⅸ chloride(CoPP)to induce heme oxygenase-1(Ho-1)upregulation in rat islet cells.Mthods Forty rats were divided into 5 groups by management 24 h before islets isolation:group A received inlzaporitoneal injection with 2.5 ms/ks CoPP,group B with 5 ms/ks CoPP,group C with 7.5 ms/kg CoPP,and group D with 10 mg/kg CoPP.In control group NS was used instead.A modified Goth approach was used for islet isolation.the yield and purity of the islets were assessed.The expression of Ho-1 mRNA and protein were detected by real time-PCR and Western blotting respectively.Islet function was tested by glucose stimulation test.Result Severe damage was found in tlle rats in group C and D.There was no difference in islet yield and purity for group A、B、C and control(P＞0.05).Group B had the highest Ho-1 mRNA and protein expression among the 4 groups.Though there was no difference in insulin secretion by low glucose challenge for group A、B and control's islets(P＞0.05),when challenged by hish level of glucose,significant deviation was observed.The imulin secretion level was(172.37±16.4)、(187.68±19.93)and(91.25±Conclusion Peritoneal iajection of 5 mg/kg CoPP can significantly enhance the expression of Ho-1 mRNA and protein in tat islet safely and enhance the function of islet when challenged by hish concentration of glucose.

y screen and prevent CAD in these people before diabetes sets in.

Objective To explore the relationship between histological chorioamnionitis(HC),fetal vasculitis(FV)and the morbidity of neonatal respiratory distress syndrome(RDS).Methods Three hundred and forty-seven cases of infants at the gestational ages of 28 +0 to 31 +6 weeks who were admitted to the Neo-natology Department of our hospital from October 2009 to June 2013 were analyzed retrospectively.They were divided into four groups according to the occurrence of HC and FV,namely,HC positive group and HC negative group,FV positive group and FV negative group.The patients in the HC positive group were further divided into FV positive group and FV negative group according to the occurrence of FV.The morbidity of RDS among above-mentioned groups was compared.Results The clinical characteristics including propor-tion of male,gestational age,birth weight,cesarean delivery,antenatal corticosteroid use,gestational hyperten-sion,gestational diabetes and cholestasis of pregnancy showed no statistically significant difference among all the groups(P 〉0.05).The incidence of RDS in the HC positive group(145 cases)was 49.6%(72 cases), which was significantly lower than that in the HC negative group(67.3%,126 /202,P 〈0.05).The incidence of RDS in FV positive group(64 cases)was 42.2%(27 cases),which was significantly lower than that in FV negative group(63.3%,179 /283,P 〈0.05)In the HC positive group(145 cases),the incidence of RDS in FV positive group (64 cases)was 42.2% (27 cases),and 55.6% (45 cases)in FV negative group (81 cases),which showed no significant difference(P ﹦0.01 ).Conclusion (1 )HC or FV can reduce the incidence rate of RDS in premature infants.(2)HC combined FV cannot furtherly reduce the incidence of RDS.

Objectlve To explore the mechanism of the protection in high expression of HO-1 induced by CoPP on murine islet xenografts. Method An islet transplantation of a SD rat-to-C57 BL/6 mouse model was established.Mice were randomized into five groups i.e.control,CoPP-induction in vivo,CoPP+ZnPP in vivo.CoPP-induction in vitro and CoPP+ZnPP in vitro and the islet xenografts were transplanted into the subrenal capsule.Normoglycemia time was recorded and insulin-releasing test was performed.IL-10、TNF-α、IL-1β and INF-γ in serum and their cytokine mRNA and HO-1 in xenografts were measured by RT-PCR and Western-blotting.The pathological examination was done to observe the lymphocyte infiltration. Results There Was significant difference in the normoglycemia time between CoPP-induction in vivo and in vitro and other three groups.The results of insulin-releasing stimulated by low level glucose were identical among groups,but that of insulin-releasing stimulated by high-glucose in in vivo group were the hiishest as in CoPP-induction in vivo and in vitro and control group were 187.68 ±19.93、137.22±11.73,91.25±12.64 μIU·ml~(-1)·10islets~(-1)·45 min~(-1),(P<0.05).The IL-10 in serum in CoPP-induction in vivo and in vitro(in vivo:72.97±9.74 pg/ml;in vitro:70.84±3.56 pg/ml)was significantly hisher than other three groups(control:30.57±3.93 pg/ml;CoPP+ZnPP in vivo:39.78±3.00 pg/ml;CoPP+ZnPP in vitro:35.42±4.30 pg/ml).The expression tendency of IL-10 mRNA was similar to that of insulin secretion.There was no significant difieFence in TNF-α、IL-1β and INF-γ.The expression of HO-1 by PCR and Western-blot analysis in CoPP-induction in vivo and vitro was higher than other three groups.The pathological examination showed that fewer lymphocytes infiltrated into the islet xenografts from CoPP-treated in comparison with xenografts from other three groups. Conclusion HO-1 could improve the survival of islet xenograft:the induetion of HO-1 expression in vivo was much mole efficient than in vitro.The CoPP-induction could be related to immune modulation of IL-10.

Objective To investigate whether cerebral microbleeds (CMBs) can predict hematoma expansion in hypertensive cerebral hemorrhage bleeding.Methods One hundred and forty-four patients with hypertensive cerebral hemorrhage bleeding in 6 hours after the onset of symptom were included.Gradient echo pulse sequence-T2 WI (GRE-T2 WI) and computed tomography (CT) were performed to detect the size of hematoma in half an hour after hospital admission.Based on the performance of GRE-T2 WI,patients were divided into microbleeds group and no microbleeds group.CT was performed 24 and 72 hours later to check whether hematoma was enlarged,the ratio of hematoma enlargement and the increased hematoma volume were compared between 2 groups.Results A variable number of CMBs were found in 74 cases by GRE-T2WI on admission.The hematoma volume was increased in 12.5％ (18/144) of patients by CT 24 hours later,and in 13.9％ (20/144) by CT 72 hours later.The ratio of CMBs in microbleeds group was higher than no microbleeds group significantly (70.0％ (14/20) vs 48.4％ (60/124),x2 =4.221,P ＜0.01).Besides,the ratio of the patients with the increased hematoma volume in microbleeds group was significantly higher than no microbleeds group(17.6％ (13/74) vs 10.0％ (7/70),x2 =3.172,P ＜ 0.05).Logistic multiple regression showed that CMBs was the only risk factor which could enter regression equation (OR=2.213,95％CI 1.320-2.972,P＜0.01).Conclusion CMBs patients with hypertensive cerebral hemorrhage bleeding in GRE-T2WI can predict the high risk of hematoma expansion.

Objective To investigate the factors which may affect the secretions of human leptin and free fatty acids (FFA) by measuring plasma glucose, serum insulin, leptin and FFA levels during reduced sample number intravenous glucose tolerance tests (IVGTT) in the subjects with different status of insulin resistance and ? cell function. Methods According to oral glucose tolerance test, 7 normal, 10 overweight/obese individuals, 12 subjects with impaired glucose tolerance (IGT) and 11 with type 2 diabetes mellitus (DM) were recruited. During IVGTT, serum insulin, leptin and FFA levels at 12 time points were simultaneously measured. Results Compared to the stable secretion of leptin, during 180 min in the IVGTT, the FFA secretion showed a "U" shape profile. The correlation analysis showed that the average serum leptin and FFA levels in 3 h were independent to fasting plasma glucose concentration. The partial correlation coefficient of leptin and insulin decreased after being adjusted for FFA (before r=0.77, P0.05). Conclusion The changes of serum FFA levels in IVGTT are more significant than those of leptin after glucose loading. Fasting plasma glucose levels do not significantly affect the average serum levels of leptin and FFA. On the contrary, insulin manifests such action but does not affect the secretion profile of these parameters.

AIM: To investigate the effects of transient receptor potential channel 6 (TRPC6) on the proliferation of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS) induced by IL-1β.METHODS: The mRNA expression of TRPC6 in synovial tissues from RA or OA patients was studied by RT-qPCR.RA-FLS were cultured by enzyme digestion and tissue adhesion methods.The method of flow cytometry was applied to identify the RA-FLS.RA-FLS were treated with different concentrations (0, 0.25, 0.5, 1, 2, 4, 8 and 16 μg/L) of IL-1β for 36 h.The cell viability was examined by CCK-8 assay.RA-FLS were incubated with IL-1β (16 μg/L) for different time (12, 24, 36, 48, 60 and 72 h), and the cell viability was measured by CCK-8 assay.The interference efficiency of TRPC6-siRNA was determined by RT-qPCR and Western blotting.After incubation in the presence or absence of IL-1β medium, the cell viability, the percentage of EdU-positive cells and the percentage of (G2/M+S) phase were measured by CCK-8 assay, EdU labeling assay and flow cytometry, respectively.RESULTS: The mRNA expression of TRPC6 was found in synovial tissue with higher levels in RA patients than that in OA patients.TRPC6-siRNA significantly decreased the mRNA and protein expression of TRPC6 (P<0.05).When RA-FLS were treated with IL-1β, the proliferation of RA-FLS was increased (P<0.05).The differences of the cell viability, the percentage of EdU-positive cells and the (G2/M+S) phase percentage between TRPC6-siRNA group and blank control group or NC-siRNA group were significant, in the presence of IL-1β (P<0.05).However, they were not significant in the absence of IL-1β.CONCLUSION: TRPC6 is involved in the proliferation of RA-FLS induced by IL-1β.Silencing of TRPC6 gene inhibits the growth of RA-FLS induced by IL-1β.

BACKGROUND:Triggering receptor expressed on myeloid cel s 2 (TREM-2) is highly expressed throughout the synovial tissue in active rheumatoid arthritis patients, but the role of TREM-2 in the pathogenesis of rheumatoid arthritis stil remains unclear.
OBJECTIVE:To explore the TREM-2 expression in the synovial tissue of col agen type II-induced arthritis rats.
METHODS:The col agen-induced arthritis models were established in rats. The activity indicators and pathological changes of arthritis synovial were dynamical y observed. The mRNA levels of TREM-2, tumor necrosis factor-α, interleukin-1β, and interleukin-10 were detected in synovial tissue of rats by RT-PCR. The protein expression and location of TREM-2 were measured with western blot assay and immunohistochemistry, respectively.
RESULTS AND CONCLUSION:At day 13 after immunization, the paws of model rats appeared red and swel ing, the arthritis index scores were increased (P<0.01). At day 19-25 after immunization, the inflammation reached the peak. Hematoxylin-eosin staining showed that, the synovium of col agen-induced arthritis rats were proliferated and were infiltrated by inflammatory cel s, cartilage was destroyed. Compared with the control group, the expression of TREM-2 mRNA and protein, the mRNA levels of tumor necrosis factor-αand interleukin-1βin synovial tissue of the model rats were significantly increased (P<0.05 or P<0.01), while interleukin-10 mRNA expression was significantly decreased (P<0.05). Experimental findings indicate that, TREM-2 is a crucial inflammatory regulator and the increasing expression of TREM-2 plays an important role in the pathogenesis of col agen-induced arthritis.

Objective To analyze the dose-effect relationship between cobalt protoporphyrin (CoPP) and heme oxygenase-1 (HO-1) expression in islets and to investigate the protective effect of strongly expression of HO-1 in islet xenotransplantation. Methods Donor islets isolated and purified from SD rats were randomly divided into 5 groups and incubated with different doses of CoPP for 24 h.Group A: 0 mmol/L; Group B: 5 mmol/L; Group C: 25 mmol/L; Group D: 50 mmol/L; Group E:75 mmol/L. The expression of HO-1 mRNA and protein in islets was detected by RT-PCR and Western blot respectively. Glucose of low and high concentrations was added to islets in vitro to test insulin-releasing function. The optimal dose of CoPP which could induce the strongest HO-1 expression was chosen according to the results. Recipients were randomly divided into 2 groups. Control group received untreated xeno-islets, and the experimental group received islets incubated with optimal CoPP close in vitro. Glycemia and rejection were observed after transplantation daily. Results The expression of HO-1 mRNA and protein in xeno-islets of group D was significantly higher than in other groups (P＜0.05). After stimulation of glucose, the insulin concentration in group D was significantly higher than in other groups (P＜0.05). The optimal dose of CoPP which could induce the strongest HO-1 expression was 50 mmol/L. The time for normoglyeemia in experimental group was (14.63±1.19) days, significantly longer than that in control group (9.88±2.17)days (P＜0.01). Conclusion The strongest expression of HO-1 induced by CoPP in vitro promotes the glucose-stimulated insulin secretion of islets and prolonged the survival of xeno-islet grafts by protecting them from rejection.

Objective To investigate the clinical and genetic characteristics in a male patient with 21hydroxylase deficiency combined with adrenal and testicular tumors.Methods Clinical features and laboratory data were collected from the patient.Testicular biopsy was performed.The CYP21 gene was sequenced for mutations.Results The patient presented left adrenal and testicular enlargements.The laboratory examinations showed that plasma ACTH,androstenedione,testosterone,progesterone,and 17-hydroxyprogesterone were markedly elevated.CT scan revealed that the right adrenal gland being resected and the left adrenal with nodular enlargement.Furthermore,testicular biopsy showed a prominent peritubular fibrosis with increased number of peritubular fibroblasts,tubular hyalinisation,and calcification.Sequencing analysis showed a A>G homozygous mutation at intron 2.Conclusion Patients with untreated 21-hydroxylage deficiency may.have adrenal adenomas and(or)testicular adrenal rest tumor simultaneously.