0.05).Frequencies of P12A12 genotype and A12 allele in DN group were significantly decreased respectively,when compared with DN-0 group(for P12A12 genotype,9.1% vs 18.1%,P=0.034,OR=0.453;for A12 allele,4.5% vs 9.0%,P=0.041,OR=0.479). Conclusion The observations suggest that P12A polymorphism of PPAR?2 gene is associated with Chinese type 2 diabetic nephropathy,and A12 allele may protect the development of diabetic nephropathy in type 2 diabetic patients of Chinese.

Objective: To explore the false-negative possibility in genetic test of congenital long QT syndrome (LQTS) by next-generation sequencing (NGS). Methods: A total of 28 genomic DNA samples were collected from 4 laboratories including 2 commercial medical laboratories using HiSeq2000 platform as Lab1,n=6 and Lab2,n=8; 1 commercial research service laboratory using Ion-torrent platform as Lab3,n=8 and 1 academic laboratory using HiSeq2000 platform as Lab 4,n=6. Sequencing coverage in the exons of protein-coding region in 3 main LQTS pathogenic genes as KCNQ1, KCNH2, SCN5A and possible pathogenic variants were quantitatively analyzed. Results: In Lab1, Lab 2 and Lab 4 with HiSeq2000 platform, above 98% protein coding regions in 3 pathogenic genes were covered with>10-fold reads and 90%-95% were covered with>30-fold reads. In 2 commercial medical laboratories, 3.63% and 9.84% protein coding regions of KCNQ1 gene in 14 samples were covered with<10-fold reads and with<30-fold reads; lower than 10-fold covering region was focused in the 1st exon including about 2% known or likely pathogenic variants. In 2 commercial medical laboratories, 2.64% and 15.76% protein coding regions of KCNH2 gene in 14 samples were covered with<10-fold reads and with<30-fold reads; low covering region was located in multiple exons. For the data from Lab 1, as high as 28.56% protein coding regions of KCNH2 gene were covered with<30-fold reads including 113 (19.79%) known or likely pathogenic variants. SCN5A gene had the best coverage of protein coding region, with no<10-fold reads in all 4 Labs and no<30-fold reads in 2 commercial medical laboratories. Conclusion: Currently, NGS has low coverage region in both KCNQ1 and KCNH2 genes, pathogenic variants could be missed and false-negative possibility should be highly alert.

Objective To explore the relationship between Arg913Gln(G→A) polymorphism of solute carrier family 12 member 3 (SLC12A3) gene and diabetic nephropathy (DN) in type 2 diabetes mellitus (T2DM) in Han population of Shanghai. Methods Two hundred and fifty-eight Han ethnic people in Shanghai with T2DM (T2DM group) were divided into non-DN group (DN0 group, n=95) and DN group (n=163) according to 24 h urine albumin excretion rate (AER), and those in DN group were subdivided into microalbuminuria group (DN1 group, n=95) and macroalbuminuria group (DN2 group, n=68). Besides, 82 people with normal results of oral glucose tolerance test (OGTT), without diabetes mellitus and nephropathy were served as controls. PCR-sequencing was used to detect the genotypes of Arg913Gln polymorphism of SLC12A3 gene. Genotypic and allelic frequencies and clinical characteristics were compared among groups. Results Three genotypes (GG, GA and AA) were detected. The frequencies of GA+AA genotype and A allele in T2DM group were higher than those in control group, while there was no significant difference between groups (P>0.05). There was no significant difference in genotypic or allelic frequencies among subgroups of T2DM group (P>0.05). The level of triglyeeride (TG), AER, level of fasting insulin (FINS) and HOMA-IR in patients with GA+AA genotype were significantly higher than those in patients with GG genotype in T2DM group (P<0.05). Conclusion Arg913Gln(G→A) polymorphism of SLC12A3 gene is not significantly associated with T2DM and DN in Han population of Shanghai. The AER of people with GA+AA genotype is significantly higher than that with GG genotype. Arg913Gln (G→A) polymorphism of SLC12A3 gene may predict the risk of increase of albuminuria in patients with T2DM in Han population of Shanghai.

ObjectiveTo apply the Schwann cell neurilemma channel(SCNC) in promoting the regeneration of injuried spinal cord.MethodsSpinal cords of rats after laminectomy at T8/9 were divided in 3 groups randomly: 10 rats were transplanted with SCNC(Group A);10 rats only made models(Group B);10 rats were as normal control group(Group C).All rats received behaviour survey and inclined plane test every week 8 weeks after operation.6 weeks after operation,they were measured with cortical somatosensory evoked potential(CSEP).And 8 weeks after operation,all specimens were studied histologically.ResultsCSEP could be recorded in group A and C,but not in group B.There were marked differenties between group A or C and group B in behavior survey,inclined plane test.Amount of nerve fibers regenerated through the injury gap in group A was significantly larger than that in group B.ConclusionSCNC could facilitate axonal regeneration and promote the repairing of injuried spinal cord.

Objective: To investigate the inhibitory effect of Rh2 on HepG2 cells and explore the underlying mechanism.Methods: We used lentivirus carrying β-catenin to infect HepG2 cell, and detected expression of β-catenin using fluorescence microscopy.The effect of Rh2 on proliferation of HepG2-β-catenin and HepG2 cells was measured by CKK-8 assay,and flow cytometry was used to detect cell cycle and apoptosis.The activity of Gsk-3βwas checked by ELISA kit.The expression of Gsk-3β,β-catenin,Bax,Bcl2,CyclinD1,MMP3 genes were measured by qRT-PCR.In order to checked the relationship between β-catenin and TCF4,CHIP assay kit was used,the expression of Bax,Bcl2,CyclinD1,MMP3 genes were measured by PCR.The expressions of Gsk-3β,β-catenin,Bax,Bcl2,CyclinD1,MMP3 proteins were examined by Western blot.Results:HepG2 cells were successfully infected by pLOV-EF1a-MCS-3FLAG-β-catenin lentivirus,named HepG2-β-catenin.CCK-8 showed that ginsenoside Rh2 could effectively inhibit the proliferation of HepG2 and HepG2-β-catenin cells in vitro,which exhibits a dose-dependent manner at range of 10-160 μmol/L Rh2.The IC50 of Rh2 exposure on HepG2 cell for 48,72 h were 100 μmol/L and 58.12 μmol/L,but the IC50 of Rh2 exposure on HepG2-β-catenin for 48,72 h were 129.2 μmol/L,83.33 μmol/L,respectively.The IC50 of Rh2 exposure on HepG2-β-catenin cell was higher than HepG2 cell, compared with HepG2 group the differences was statistically significant ( P<0.01 ).Flow cytometry indicated that Rh2 could arrest HepG2 and HepG2-β-catenin cells in G0/G1 phase;the cell population in G0/G1 phase of HepG2+Rh2 group was(64.57±0.65)%,HepG2-β-catenin+Rh2 group was(58.61±2.01)%.Flow cytometry indicated that Rh2 could induced early apoptosis in HepG2 and HepG2-β-catenin cells.The apoptosis rate of HepG2 +Rh2 group was (17.27 ±2.77)%,HepG2-β-catenin +Rh2 group(9.02 ±1.76)%.The ELISA results indicated that HepG2 cells was induced by Rh2 for 12,24,48,72 h,the activity of Gsk-3βgradually increased,peak in 48 h,then decreased.Compared with control group,Rh2 induced HepG2 and HepG2-β-catenin cells for 48 hours, Gsk-3βactivity were increased, and their activity reduced after adding Bio, there were no significant differences between HepG2+Rh2 and HepG2-β-catenin+Rh2 groups.The PCR,CHIP and WB results showed that the expression of Gsk-3β,Bax gene and proteins increased,while theβ-catenin,CyclinD1,Bcl2,MMP3 gene and proteins down-regulation in HepG2 and HepG2-β-catenin cell induced by Rh2.Compared with HepG2-β-catenin +Rh2 group, the expression of other gene and proteins changed significantly,however,Gsk-3βwas no significant difference.Conclusion:Over-expression of β-catenin may weaken the phar-macological effects of ginsenoside Rh2 on HepG2 cells.The activity of Gsk-3βwas increased by ginsenoside Rh2 to degradeβ-catenin, affecting the expression of downstream genes,promoting apoptosis of liver cancer cells and inhibiting metastasis.

Objective:To study the mechanism of ginsenoside Rh2 inhibiting HepG2 cells migration.Methods:HepG2 cells in logarithmic growth phase were cultured in 96-well plates,which were induced by different concentration Rh2,respectively for 24,48,72 hours.The cell inhibition was detected by Cell Counting Kit.Transwell chambers was used to checked HepG2 cell migration ability;luciferase was tested by Luciferase Reporter Assay system reagent;The expressions of P-ERK,ERK,P-P38,P-38,P-JUK,JUK,MMP3 proteins were detect by Western blot;the expression of AP1,MMP3 gene were detected by Quantitative PCR;The expression of AP1, MMP3 fluorescence protein were observed by fluorescence microscopy.Results:Administrated with different concentration of Rh2 after 24 ,48 ,72 h,the proliferation of HepG2 cells were inhibited ( P<0.05) ,and in dose-and time-dependent manner.Transwell assay showed Rh2 could significantly inhibited migration of HepG2 cells.The expressions of P-ERK , MMP3 proteins were significantly decreased,the expressions of P-JUK, P-P38 proteins were significantly increased, expression levels of ERK, P-38, JUK were no significant difference.Expression of AP1,MMP3 gene were significantly decreased,the expressions of AP1,MMP3 fluorescence proteins were significantly decreased.Conclusion:Ginsenoside Rh2 can activate MAPK pathway to inhibit the migration of HepG2 cells.

Objective To explore the relationships of serum leptin(LP),tumor necrosis factor-?(TNF-?)and insulin resistance in simple obesity children.Methods Forty-seven children with simple obesity were divided into two groups: high insulin group(HIG) and normal insulin group(NIG) according to the levels of fasting state of serum insulin and glucose.The fasting state of serum insulin(FINS),LP,TNF-? and fasting blood glucose(FBG) were measured.The relationships of LP,TNF-? and insulin resistance index(HOMA-IR) were also analyzed.Results 1.The BMI,waist,hip,waist hip ratio(WHR),LP,TNF-?,HOMA-IR were obviously higher in the HIG than those in the NIG and NCG groups(Pa0.05).3.In the HIG group,the FINS was positively correlated to LP,TNF-? and HOMA-IR(r=0.560,0.413,0.864 P

Acetyl-CoA Carboxylase ; metabolism ; Cell Line ; Curcumin ; pharmacology ; Hepatocytes ; drug effects ; metabolism ; Humans

UNLABELLEDTo investigate the effects of Shizhang Cataplasm (SC) and Xuzhang Cataplasm (XC) in treating liver cirrhosis caused ascites of excessive syndrome (ES) type and deficient syndrome (DS) type respectively.

METHODSAll the 77 patients (37 of ES type and 40 of DS type) enrolled were treated by conventional treatment but with restrictive use of diuretics. SC and XC were given respectively to 26 patients of ES type and 26 of DS type additionally by umbilical sticking, they were regarded as the treated group, and those (11 of ES type and 14 of DS type) not received the cataplasm treatment were regarded as the control group. The changes of symptoms, body weight, abdominal perimeter and amount of urine before and after treatment were observed, and amount of ascites was examined with B-ultrasound to evaluate the efficacy according to comprehensive grading criteria. Also, the toxicity was observed.

RESULTSSixty-two cases completed the full course, 15 were withdrawn. As compared with the corresponding control group, body weight, abdominal perimeter and amount of ascites decreased, while amount of urine and flatus discharging increased remarkably in the treated group (P < 0.05). The comprehensive efficacy in patients of ES type was better than that in DS type (P < 0.05). The effective rate of grade I/II was 7.1% and 9.1% for patients in the control group of DS type and ES type respectively, while it was 57.2% and 69.2% in the treated group of DS and ES type respectively. Better therapeutic effect was shown in patients of ES type treated with SC.

CONCLUSIONSC and XC showed good assistant effects in treating patients with liver cirrhosis caused ascites of ES and DS type respectively.

Administration, Cutaneous ; Ascites ; diagnostic imaging ; drug therapy ; etiology ; Diagnosis, Differential ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Liver Cirrhosis ; complications ; diagnostic imaging ; drug therapy ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy ; Ultrasonography ; Umbilicus

China ; Heart Transplantation ; methods ; Humans