The study of subjcts with T2DM (n=120) and NGT (n=106) in using the screening of PGC-1? gene for SNPs by PCR-SSCP and DNA sequencing showed that four SNPs(Thr394Thr、Gly482Ser、Thr528Thr、Thr612Met) in the PGC-1? gene were found in 50 Han Chinese type 2 diabetic patients.A Gly482Ser polymorphism of PGC-1? gene was associated with type 2 diabetes.[

Objective To evaluate surgical therapy for recurrent rectovaginal fistula.Methods In this study.two patients were treated by endorectal advancement flap repair and one patient was treated by vascular pedicled segment of small bowel.Results All patients were cured and followed up from 4 to 20 months.During the period of follow-up there was no recurrence.Conclusions The procedures and timing of operation are important factors for a successful repair.Both the endorectal advancement flap and patch of intestine provide an effective methods in repairing recurrent rectovaginal fistula.

Objective: To investigate the antitumor activities and apoptosis mechanisms of 3,7-dinitrodibenzobromonium salt (cBr) on human chronic myelogenous leukemia K562 cell line in vitro. Methods: The antitumor activities of cbr were tested by Trypanblau exclusion test, MTF coloretric assay and clonal forming test, and the mechanisms were studied by morphological analysis (microscopy, fluoreseencemicroscopy, electron microscopy) and flow cytometry analysis. Resuits: cBr could inhibit of the proliferation K562 cells and kill them significantly. The effects varied with different time and doses, and IC50 of cell colony forming rate was 0. 497 mmoL/L. Apoptosis rate tested by flow cytometry was 70.34%,and apoptotic characteristics could be found by microscopy and electron microscopy. Conclusion: cBr could inhibit K562 cell proliferation greatly, and could induce K562 cells into apoptosis.

Objective To investigate the correlation of polymorphism of 8473 (T/C,rs5275) in the 3′-untranslated region of cyclooxygenase-2 (COX-2) gene with bladder cancer. Methods A case-control study on the relation between COX-2 polymorphism and bladder cancer was performed in this study. The Taqman SNP genotyping assay was used to study the COX-2 rs5275 polymorphism. Results The differences in allele or genotype distributions of COX-2 rs5275 between cases and controls (all P < 0.05) were significant. A significantly reduced risk was revealed in bladder cancer patients carrying the TC genotype (adjusted OR =0.178;95%CI:0.119 ~0.264),CC genotype (adjusted OR = 0.087; 95%CI:0.058 ~ 0.129) or (TC/CC) genotype (adjusted OR =0.122;95%CI:0.082~0.181) compared to the control group. Significant difference in genotype distribution of the COX-2 rs5275 site was found associated with sex and smoking (adjusted OR:2.125,0.476;95%CI:1.500 ~3.010,0.325 ~ 0.696);No corelation was found between genetype TT or TC/CC and the pathological features of bladder cancer (P>0.05). Conlusion The separate effect of rs5275 polymorphism of COX-2 is associated with the susceptibility of bladder cancer, the TC/CC genotypes may be a protective factor.

To look for a more stable and convenient way of constructing short hairpin RNA expression vectors targeting the latent membrane protein-1(LMP-1) encoded by Epstein-Barr virus(pshLMP1), and to study the inhibition function of pshLMP1 expression vectors in HNE1 cells, we designed the pshLMP1 expression cassette and pshLMP1 expression vectors by both the annealing method and PCR method and then co-transfected with pEGFP-N1-1158 into HNE1 cells to observe the mRNA and protein levels of LMP-1 genes by green fluorescence analysis, RT-PCR and western blot. pshLMP1 expression vectors were successfully obtained by both methods but better cloning efficiency was achieved and fewer deletions and mutations of nucleotides were achieved with the PCR method. Furthermore, the mRNA and protein levels of LMP-1 genes were down-regulated by pshLMP1 expression vectors. According to our research, we found that the PCR method provides a more efficient way to construct pshLMP1 expression vectors which have the ability to inhibit the function of LMP-1 genes expressed in HNE1 cells, and also provides a novel application of RNA interference technology against-EBV.

A fragment sized 400bp of White spot syndrome virus（WS SV，formerly de signated NOSV），recovered from recombinant plasmid pAFD, was labeled with Digox igenin as a probe to detect dynamic distribution of WSSV within 120h and 72h in crawfishes（Cambarus proclarkii） inoculated WSSV by oral taking and injecti on r espectively. Stomach epithelium, intestine epithelium, heart, gill, haemolymph, muscle, hepatopancreas, hypoderm, connective tissue and ovary of infected crawfi shes were examined for WSSV. In both groups, WSSV was first detected in heamoly mph at 12h p.i. and then disappeared. Again it was detected at 96h p.i. only in oral infection group and maintained till 120h p.i., but it didn't appear at 72h p.i. in injection group. WSSV in heart, muscle was detected at 36h p.i. in oral infection group and 24h p.i. in injection group respectively, and then increased generally. In addition, WSSV in intestine epithelium, connective tissue, ovary of oral infection group and intestine epithelium, hypoderm, ovary of injection g roup could also be detected. In dead crawfishes after 120h and 72h p.i. in two groups, WSSV could be detected in all the examined tissues and it demonstrated t hat systemic infection occurred in the animales. The tissue containing more amo unts of WSSV was hypoderm in oral infection group, while intestine epithelium, g ill, hypoderm, ovary in injection infection group. It deduced that WSSV first a ppears in haemolymph and then goes into heart, muscle and other tissues and prol iferates in them. Once again, WSSV is released into heamolymph resulting in syst emic infection till crawfishes' death.

Objective To comparative analysis the rabbit VX2 tumors angiogenesis in fatty liver and normal liver,and investigate the correlation of contrast-enhanced ultrasound (CEUS) parameters and their angiogenesis.Methods Rabbit models of fatty liver and normal liver with implanted VX2 tumors were established.Two groups of hepatic backgrounds and VX2 tumors were analyzed by QontraXt quantitative software of CEUS.The expression level of tumor microvascular density (MVD) and vascular endothelial growth factor (VEGF) were detected with immunohistochemical techniques and real-time fluorescence quantitative PCR (FQ-PCR).Results ①No significant difference was found in MVD,VEGF protein and gene expression between VX2 tumors in normal liver and fatty liver.The expression of VEGF gene in fatty liver parenchyma were lower than normal liver parenchyma (P ＜0.05).②There was positive correlation between MVD and peak intensity (r =0.494,P ＜0.05; r =0.591,P ＜0.01) both in fatty liver and normal liver VX2 tumors.The expression of VEGF protein were not correlated with TIC parameters (all P ＞ 0.05).③The MVD had positive correlation with expression of VEGF protein in fatty liver and normal liver VX2 tumors (r =0.508,P ＜0.05; r =0.570,P ＜0.01).Conclusions The expression of MVD and VEGF had no significant difference between fatty liver and normal liver VX2 tumors.The peak intensity of VX2 tumors CEUS had positive correlation with MVD both in fatty liver and normal liver,which can indirectly reflect MVD expression level and help to evaluate tumor angiogenesis.

Objective To investigate the conventional ultrasound characteristics of VX2 tumors in fatty liver and normal liver of rabbits,furthermore,to analysis the blood flow characteristics of them with contrast-enhanced ultrasound (CEUS).Methods Rabbit model of fatty liver and normal liver with implanted VX2 tumors were established,and examined using conventional and CEUS.QontraXt quantitative software of CEUS was used.Results The fatty liver models with 20 VX2 tumors and normal liver models with 23 VX2 tumors were successfully developed in rabbits.In conventional ultrasonography,hyperechoic VX2 tumor lesions in fatty liver were far less than them in normal liver (P =0.008).No significant difference was found in iso-echoic and hypoechoic VX2 tumor lesions between two groups (P ＞ 0.05).Compared with normal liver parenchyma,arrival time of fatty liver parenchyma was later,peak intensity was lower (P ＜0.05).There was no significant difference in arrival time,peak intensity,sharpness,area under curve between the VX2 tumors of normal liver and fatty liver (P ＞ 0.05).Conclusions There is a significant difference characteristic in conventional ultrasonography between VX2 tumors in fatty liver and normal liver.Fatty liver blood perfusion effects are lower than normal liver in CEUS.Quantitative parameters of CEUS are more objective and accurate for hemodynamic of VX2 tumors and fatty liver.

Objective To analyze hepatic inflammatory pseudoneoplasm with fatty liver and normal liver perfusion blood characteristics and quantitative parameters using real-time contrast-enhanced ultrasound (CEUS) and time intensity curve.Methods Twenty rabbits (including ten with fatty liver and ten with normal liver) with thirty-nine inflammatory pseudoneoplasm of the liver (IPL) were studied by low mechanical index CEUS with bolus injection of SonoVue via peripheral auricular vein and QontraXt quantitative analysis software.IPL contrast-enhanced ultrasound blood perfusion characteristics with two different liver background were analyzed.Results ①According to the different pathological elements,IPL showed four kinds of CEUS types:one lesion showed absent contrast enhancement pattern (type Ⅰ),nineteen lesions showed iso-enhancement pattern (type Ⅱ),six lesions showed hypo-enhancement pattern (type Ⅲ),thirteen lesions showed fast-in and fast-out pattern (type Ⅳ).②Compared with normal liver,arrival time of fatty liver was later and its peak intensity was lower.②Compared with peripheral parenchyma,arrival time of IPL was later,peak intensity was lower and its sharpness was sharper (P ＜0.05).④Between IPL lesions with two different liver background,there was no significant difference about parameters of the time-intensity curve (P ＞0.05).Conclusions Due to different pathological elements,IPL lesions have multiform perfusion patterns in CEUS.CEUS and time intensity curve could provide enough perfusion information for IPL and liver parenchyma more accurately.

OBJECTIVETo observe the mucosal immune mechanism of anti-tumor action of Ganoderma lucidum polysaccharides (GLP).

METHODSThe concentration of H22 cells in suspension were adjusted to 1 x 10(9)/ L, and 0.2 mL of the cell suspension was injected subcutaneously in the right oxter of Kunming mice. Then the H22 bearing mice were randomly divided into 4 groups: the GLP group, the Cytoxan (CTX) group, the CTX + GLP group and the untreated model group, 8 mice in each group. Besides, a blank control group was set up. Starting from the 2nd day of modeling, GLP, at the dose of 1.02 g/kg was given to GLP group and GLP + CTX group by gastrogavage once a day for 12 successive days; CTX at the dose of 100 mg/kg was administered via peritoneal injection to the CTX group and the GLP + CTX group on the 1st day and the 6th day of the experimental course; but to the model group and the blank group, only equal volume of distilled water was given. All mice were sacrificed on the 14th day, the ileum at 1 cm upper to cecum was taken, through 4% paraform fixation and paraffin section, it was used for immunohistochemical detecting expressions of immunoglobulin A (IgA), tumor necrosis factor-alpha (TNF-alpha), interleukin-2 (IL-2) and interleukin-10 (IL-10) in ileum. Besides, the lymphocyte subsets in the intraepithelial lymphocyte (IEL), lamina propria lymphocytes (LPL), and Peyer's patch lymphocytes (PPL) were analyzed by immune fluorescence technique and flow cytometry.

RESULTSCompared with the blank control group, the phenotype of lymphocytes and the expression of cytokines in ileum in the model group changed significantly; and the phenotype was variant in different regions. Compared with the model group, both indexes were adjusted in the GLP, CTX and GLP + CTX group to different degrees.

CONCLUSIONThe adjustment of GLP on intestinal mucosal immune is probably another path for its anti-tumor action.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Immunoglobulin A ; metabolism ; Interleukin-2 ; metabolism ; Intestinal Mucosa ; immunology ; Liver Neoplasms, Experimental ; immunology ; Male ; Mice ; Mice, Inbred Strains ; Polysaccharides ; isolation & purification ; pharmacology ; Random Allocation ; Reishi ; chemistry ; Tumor Necrosis Factor-alpha ; metabolism