Objective To study the mechanism of insulin-stimulated glucose uptake in skeletal muscle cells. Methods The responses of GluT4 translocation and glucose uptake to the investigational drugs of SB203580 and Wortmannin as well as the effect of insulin on the drugs during differentiation were detected to study the insulin signal pathway. Results The GluT4 translocation and glucose transport were increased under insulin stimulation by 2.5±0.2 and 2.2±0.1 folds, respectively while compared with control; but t1/2 were 3.3 min and 6.0 min, and IC50 to wortmannin were 43 nmol/L and 3 nmol/L respectively.SB203580 inhibited 64% and 62% of insulin-stimulated glucose uptake and photolabelling of cell surface GluT4, respectively, but had no effect on GluT4 translocation.The fold increase of insulin-stimulated glucose uptake (1.7±0.1 fold vs control)was lower than that of GluT4 translocation (2.3±0.1 fold vs control) in myoblasts. Conclusions In skeletal muscle cells, two insulin signal pathways mediate GluT4 translocation and activation of GluT4, respectively.Insulin engages both of the pathways to stimulate the cells for maximum glucose uptake.

Objective To observe the effects of berberine on inflammatory factors, adipokines and fatty acid metabo-lism in 3T3-L1 adipocytes, and to investigate the molecular mechanism underlying berberine’s role of improving insulin re-sistance. Methods mRNA level of inflammatory molecules, adipokines, key enzymes and protein in fatty acid metabolism in 3T3-L1 cells were determined by quantitative real time polymerase chain reaction (qRT-PCR) after cells were treated with different concentrations of berberine (0, 5, 10, 20, 40μmol/L) for 24 hours and with 10μmol/L berberine at different du-rations (0,4,8,24,48 h). These factors mainly included interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α), leptin, adipo-nectin, visfatin, fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), adipose triglyceride lipase (ATGL) and adipocyte fatty acid binding protein (AFABP). Results In 3T3-L1 adipocytes, transcription level of IL-6, TNF-α, leptin, FAS, AT-GL, AFABP reduced with addition of berberine dosage at 10~40μmol/L(P＜0.05)while visfatin mRNA level increased(P＜0.05)compared with the control group. No significant difference was found in expression of adiponectin(P>0.05). Tran-scription level of IL-6, TNF-α, leptin, AFABP, ATGL, FAS decreased with time after 10μmol/L berberine intervention (8-48 h) compared with the control group(P＜0.05). On the other hand, visfatin mRNA level increased(P＜0.05)compared with the control group. Adiponectin mRNA decreased only after cells were treated with berberine for 48 h(P＜0.05). No sig-nificant difference was found transcription of ACC between each groups treated with berberine(P > 0.05). Conclusion mRNA level of inflammatory factors, adipokines, key enzymes and protein in fatty acid metabolism in 3T3-L1 adipocytes can be affected by berberine and this effect depend on its dose and time . This might be the mechanisms underlying berber- ine to improve insulin resistance.

Objective To determine levels of nuclear factor (NF)-κB, interleukin (IL)-10, and visfatin in adipocytes treated by different degrees of intermittent hypoxia (IH), and to investigate the mechanism of IH leading to insulin resistance (IR). Methods The cell model of intermittent hypoxia/re-oxygenation (IH/ROX) in obstructive sleep apnea (OSA) was established. Differentiation mature 3T3-L1 adipocytes, were randomly divided into 10 groups including four different-frequency intermittent hypoxia groups(IH1-4, fixed intermittent hypoxia scheme for 1.5%O2 45 s and then re-oxygen 21%O2 for 2 min 15 s, 4 min 15 s, 5 min 45 s and 8 min 45 s, 60 times circulation), and their normal oxygen control groups (SC1-4, instead each IH group 1.5%O2 to 21%O2, the rest groups were treated as same as IH group), continuous hypoxia group (CH, 10%O2 for 6 h) and normal oxygen control group (CC, 21%O2 for 6 h). ELISA method was used to determine the levels of IL-10 and visfatin in the supematant of adipocytes. Western blot method was used to determine the protein levels of NF-κB p65 and visfatin. Real-time PCR method was used to determine the mRNA levels of IL-10 and visfatin. Results The protein and mRNA expressions of IL-10 were significantly lower in IH group and CH group than those of control groups (P<0.01). The levels of NF-κB p65 protein were significantly increased in IH group and CH group than those of control group. The protein and mRNA expressions of visfatin were significantly higher in IH1, IH2 and CH groups than those of control group (P<0.01). Conclusion As a prominent feature of OSA pathophysiology, IH may take part in insulin resistance of OSA patients by abnormally secreting NF-κB, IL-10 and visfatin in adipocytes.

AIM:To observe the effect of the extract of Ginkgo biloba(EGB)on pituitary-testicular axis and the mRNA expressions of luteinizing hormone receptor(LHR)and steroidogenic acute regulatory protein(StAR).METHODS:Thirty male Sprague-Dauley rats were divided into three groups randomly:normal control group,type II diabetic group and EGB treatment group.After fed with high-fat diet for 4 weeks,the later two groups were injected with strepozotocin intraperitoneally to induce type II diabetes mellitus.The EGB treatment group was given EGB at the dose of 50 mg/kg once a day for 12 weeks by intragastric administration.The normal control and diabetic group were given normal saline of equal volume per day for 12 weeks.The indices of blood glucose,insulin and low-density lipoprotein-cholesterol(LDL-c)were measured.The morphologic change of testicular tissue was observed under light microscopy(LM)and transmission electron microscopy(TEM)respectively.The concentrations of blood luteinizing hormone(LH)and testosterone(T)were assayed by the technique of enzyme linked immunosorbent assay(ELISA).The mRNA expressions of LHR and StAR from Leydig cells were detected by RT-PCR.RESULTS:The concentrations of blood glucose,insulin and LDL-c increased obviously,and the testis weights lessened obviously in type II diabetic groups compared to those in normal control groups.Rare spermatogenic cells of seminiferous tubule and germinal arrest were observed in diabetic group under LM.Ultrastructural analysis of testicular tissue by TEM showed dilation of the endoplasmic reticulum and mitochondrial swelling in Leydig cell and sertoli cell in diabetic group.The level of blood LH and T decreased in typeⅡ diabetic groups in comparison with that in the normal control group.Compared to normal groups,the mRNA expression of StAR in type Ⅱ diabetic groups decreased,while the mRNA expression of LHR increased.After the treatment of EGB,the pathological change of testis was relieved,the concentrations of blood glucose,insulin and LDL-c were decreased,the level of blood LH and T,and the mRNA expression of StAR were increased,and the mRNA expression of LHR descended compared to type Ⅱ diabetic groups.CONCLUSION:EGB may increase the LH-induced testosterone production by correcting metabolic disorder of glucose and lipid,improving the function of pituitary-testicular axis and regulating the expression of LHR and StAR mRNA.

Objective To establish homogeneous immunoassay for detecting serum cardiac troponin I (cTnI) by using light induced chemiluminescent immunoassay (LiCA). Methods Polyclonal antibodies of cTnI were coated on the receptor particles, monoclonal antibodies of cTnI were biotinylated, and the donor particles were coated with streptavidin, all of which were composed of LiCA reagents. The optimal test conditions and analytical performance of the detection method were studied. Results The method was rapid, sensitive, and detection time was 17.5 min.The analytical sensitivity was 0.045 ng/mL and the functional sensitivity was 0.053 ng/mL.The recovery rate was 104.96%-108.21%;The within-run and the between-run coefficients of variation were 3.88%-5.53%and 7.60%-8.75%, respectively. The interference rates for the endogenous substances were less than 10%. The reference value of cTnI was less than 1.05 ng/mL;Results of cTnI LiCA correlated well with direct chemiluminescence detection (r2 =0.979). Conclusions This approach can be used for the quantitative detection of serum cTnI, and it is homogeneous and is free of clean separation. It provides a convenient, highly sensitive detection platform for clinical practice.

Objective To examine the expression of response gene to complement 32 (RGC-32) in renal tissue of children with IgA nephropathy (IgAN), and to explore its significance. Methods The subjects were 45 children diagnosed as IgAN by renal biopsy. The expression of RGC-32, α-smooth muscle actin (α-SMA) and transforming growth factor β1 (TGF-β1) was examined by immunohistochemistry staining. The correlation of RGC-32 expression with α-SMA,TGF-β1, degree of renal pathological lesions and clinical index in IgAN was assessed by Spearman correlation analysis. Results RGC-32 protein located in renal tubular epithelial cells in normal and IgAN renal tissues. The positive expression index of RGC-32 in nomal group, IgAN mild group, moderate group and severe group was (18.29±6.22)%, (23.90±9.65)%, (31.23±9.86)%,and (34.52±10.63)% respectively. With more severity of renal pathological lesions, the expression of RGC-32 in IgAN was enhanced. The RGC-32 expression was positively correlated with the score of glomerulus and renal interstitium in children with IgAN (r=0.385, 0.347, P＜0.05), as well as α-SMA, TGF-β1 (r=0.594, 0.521, P＜0.01), but was not correlated with Scr, urinary NAG/Cr,Alb/Cr, IgG/Cr, and α1-M/Cr (r =0.117, -0.115, -0.138, -0.176, -0.028, all P ＞0.05).Conclusions RGC-32 protein locates in renal tubular epithelial cells in normal and IgAN renal tissues. RGC-32 may participate in the course of renal tubulointerstitial lesions in children with IgAN, especially in the course of epithelial-mesenchymal transition (EMT) induced by TGF-β1.

OBJECTIVETo analyze the clinical features and gene mutation of Chinese children with Alport syndrome(AS).

METHODFrom May 2011 to May 2014, clinical and pathological information gathered from 25 patients was retrospectively analyzed. COL4A5, COL4A4 and COL4A3 genes were analyzed using next-generation sequencing in these patients, and gene mutations of related family members were identified by Sanger method.

RESULTOf these 25 cases, 19(76%) had X-linked Alport syndromes (XL-AS), 6 had autosomal recessive Alport syndromes (AR-AS). Twenty five patients had an onset of hematuria and proteinuria and in 8 cases the disease was induced by upper respiratory tract infections. Hearing loss was present in 2 of 25 (8%) cases and ocular lesions in 1 of 25 (4%). Renal pathology showed that 16 of them had minimal change disease (MCD), 8 mesangial proliferative glomerulonephritis (MsPNG), 1 focal segmental glomerulo-sclerosis (FSGS). Extensive lamination and split of glomerular basement membrane (GBM) dense layers were found in 2 (8%) of 25 patients. Twenty one of 25 patients (84%) showed abnormal renal α-chain distribution. COL4A5, COL4A4 and COL4A3 genes of 25 patients (23 families) were analyzed and 24 pathogenic mutations were identified: 18 in COL4A5, 1 in COL4A3 and 5 in COL4A4. It was observed that 13 patients inherited the mutation from the mother, 3 patients inherited from the father, 2 patients inherited 1 mutation from the mother and another mutation from the father, and 7 patients carried the novel mutations.

CONCLUSIONXL is the main inherited type in AS. Most of patients showed MCD and MsPNG in renal biopsy. This research examined 24 mutations and 16 mutations were not reported previously.

Child ; Deafness ; Genes, Recessive ; Genotype ; Hematuria ; Humans ; Kidney ; Mutation ; Nephritis, Hereditary ; genetics ; pathology ; Pedigree ; Phenotype

Monitoring blood glucose level timely and accurately and management of glucose level is crucial for the treatment of diabetes and the prevention of diabetic complications. Glycated albumin is one of the important biomarker to evaluate the fluctuation of blood glucose level, which has been widely used in clinic. To understand the advantages and limitations of the glycated albumin in monitoring glucose level will contribute to its better application in clinic.

Sj?gren′s syndrome(SS) is a chronic inflammatory autoimmune disease, including primary SS (pSS) and secondary SS (sSS). Pediatric sSS has the similar clinical characters with pSS, which can be classified based on the same classification criteria.Compared with those of adult SS, pediatric SS is rare, insidious and lacks the typical manifestations of dry eyes and dry mouth.Therefore, the classification criteria for adult SS is not suitable for pediatric SS.So far, there are no classification criteria for pediatric SS that have been widely applied and validated, which needs to be further explored.The present review described the clinical characteristics and progress of classification criteria for pediatric SS, and compared pSS and sSS in affected children.

Objective To evaluate the clinical and esophageal dynamics characteristics, and initial treatment effect of patients with untreated achalasia using high resolution manometry. Methods From January 2012 to June 2016, patients newly diagnosed with achalasia were retrospectively recruited. Clinical information, endoscopy and X-ray findings, manometric variables and treatment outcomes were collected and compared among manometric subtypes. Results Among 133 patients with achalasia, 32 ( 24. 0%) were classified as type Ⅰ, 84 ( 63. 2%) as typeⅡ and 17 ( 12. 8%) as typeⅢ. Compared with the typeⅢ, the symptom of regurgitation was more common in the types I and typeⅡachalasia patients ( P=0. 020) . While compared with the type Ⅰ, the score of chest pain was higher in the patients with type Ⅱ and type Ⅲ ( P=0. 006) , as well as the basal lower esophageal sphincter pressure ( P=0. 015, P=0. 023 for type Ⅱ and type Ⅲ, respectively) . Integrated relaxation pressure was similar among the 3 subtypes ( P=0. 158) . Forty-five and 47 patients received pneumatic dilation ( PD ) and peroral endoscopic myotomy ( POEM ) respectively, and all of the patients were followed-up for at least 6 months. The success rate was higher in the POEM group than that of the PD group in all the 3 manometric subtypes, but only the difference between POEM and PD in the type Ⅲ was statistically significant ( 90. 0% VS 40. 0%, P = 0. 041 ) . Conclusion The clinical characteristics and treatment response are different among the 3 achalasia subtypes. Patients with type Ⅲ achalasia are probably the best candidates for POEM.