Objective To reproduce the inflammatory bowel disease(IBD) models induced by 2,4,6-trinitrobenzene sulfonic acid(TNBS),and to investigate the effect of Changkangyin(CKY) on it.Methods Total 60 rats were divided into 6 groups by random:alcohol control group,model group,salicylazosulfapyridine(SASP)group,low,middle and high dosages CKY groups.IBD model was induced by TNBS.The changes of macroscopic morphous and pathohistology of colon were observed after intervened with drug for 21 d.Results The body weight in model group was obviously lower than that in alcohol control group(P0.05).Conclusion Colitis can be induced by TNBS in rats.The change of it is the same as human's.It is a ideal model to study pathogenesis of IBD and investigate the effect of medicine.CKY has a batter therapeutic effect on IBD model.

Hemifacial Spasm ; etiology ; surgery ; Humans ; Trigeminal Neuralgia ; etiology ; surgery

It is a technological difficulty to acquire data from ultrasound with ultra-band in the study of microwave-induced thermoacoustic tomography. A high-speed data acquisition system for the ultrasonic signal with a frequency fewer than 500 MHz is designed in the paper. The result obtained demonstrates that the system holds good for microwave-induced thermoacoustic tomography.

Objective To investigate the efficacy of liver transplantation for the treatment of polycystic liver disease.Methods The clinical data of 8 patients with polycystic liver disease who received liver transplantation at the Third Affiliated Hospital from September 2003 to July 2009 were retrospectively analyzed.All patients underwent modified piggyback liver transplantation with vena cava plasty,and 1 of the patients received simultaneons liver-kidney transplantation.Data including acute rejections,complications,graft functions and survival time of patients were recorded.Results Operation was successfully performed on all patients.The mean operation time,anhepatic phase and operative blood loss were (7.5 +2.8)hours (range,4-11 hours),(42 + 14)minutes (range,25-70 minutes) and (2250 ± 1850)ml (range,2000-6500 ml),respectively.One patient received liver retransplantation at 45 days after primary liver transplantation because of hepatic artery stenosis.Two patients died during perioperative period.One patient died of pulmonary infection and multiple organ disfunction syndrome (MODS) at 39 days after operation,and the other one died of MODS at 59 days after operation.The median follow-up time was 60 months (range,37-93 months).Six patients survived for more than 3 years,4 patients survived for more than 5 years and 2 patients survived for more than 7 years.No acute rejection of gratis was observed at the end of the follow up.Conclusion Liver transplantation is safe and effective for the treatment of polycystic liver disease.

Objective To evaluate the efficacy and indications of salvage liver transplantation for patients with recurrent hepatecellular carcinoma(HCC)after liver resection.MethodsThe clinical data of 35 HCC patients who received salvage liver transplantation after liver resection at the Third Affiliated Hospital of Sun Yatsen University from October 2003 to March 2006 were retrospectively analyzed.All patients were divided into the salvage liver transplantation(SLT)group(n = 19)and extended SLT group(n = 16).Perioperative condition,postoperative complications and prognosis of the 2 groups were compared.The survival rate was analyzed and compared by the Kaplan-Meier method and Log-rank test,respectively.Results The anhepatic phase,ischaemic time,operation time,intraoperative blood loss,packed red blood cell transfusion,fresh frozen plasms transfusion,mobidity and retransplantation rate were(32 ± 9)minutes,(8.0 ± 2.1)hours,(7.6 ± 1.5)hours,2300 ml,8 U,23 U,6/19 and 2/19 in the SLT group,and(34 ± 7)minutes,(7.4 ± 2.3)hours,(7.4 ± 2.0)hours,2750 ml,12 U,20 U,4/16,1/16 in the extended SLT group,respectively,with no significant difference between the 2 groups(t=0.726,-0.804,-0.366,Z=-0.348,-0.549,-0.149,x2 =0.184,0.203,P＞0.05).The perioperative mortality,tumor recurrence rate were 0 and 2/19 in the SLT group,and 4/16 and 9/16 in the extended SLT group,with significant differences between the 2 groups(x2 = 5.363,8.426,P ＜ 0.05).The 1-,3-,5-year cumulative survival rates were 100%,84% and 84% in the SLT group,and 75%,33% and 33% in the extended SLT group.The 1-,3-,5-year tumor-free survival rates were 100%,89% and 89% in the SLT group,and 48%,29% and 19% in the extended SLT group.There were significant differences in the cumulative and tumor-free survival rates between the2 groups(x2 =11.58,19.31,P＜0.05).Conclusions The efficacy of SLT is satisfactory in the treatment of recurrent HCC.The optimal indication for SLT is Milan criteria.

Objective To investigate the effects of recombinant human thioredox ( rhTrx) on primary cultured hepatocytes. Methods The cDNA coding hTrx was amplified by using RT-PCR. The objective protein was expressed in E. Coli. The activity of purified rhTrx protein was determined by IgM and insulin disulfide reduction assay. DNA synthesis was determined by 3H-TdR mixing assay to realize the proliferating roles of rhTrx in murine primary cultured hepatocytes. Cell viability was assessed by lactate dehydrogenase ( LDH) releasing assay. Results The cloning hTrx open reading frames cDNA was confirmed by sequence analysis. The soluble recombinant hTrx are expressed efficiently in the prokaryotic expression system. The recovery ratio was 23. 20% of total bacteria protein with a purity of the expressive protein of 96. 30%. The rhTrx had stronger IgM and insulin reduced activity compared with the controls (P

Objective To identify PDZ domain containing proteins interacting with PTEN and its characterization with NHERF-1 by proteomic analysis. Methods The interactions between PTEN and PDZ domain containing proteins were screened with PDZ protein array, and the novel one was then identified with GST pull-down and co-immunoprecipitation assay. Results Using a PDZ protein array, we proved PTEN binding with NHERF-1. The interaction of PTEN and NHERF-1 was further characterized by GST pull down assay, and we demonstrated that PTEN associated with NHERF-1 via the binding of PTEN carboxyl-terminal with the PDZ domain 1 (PDZ1) of NHERF-1. The last four amino acids (I-T-K-V) of the PTEN were the key determinants of this interaction as mutation of any of the four amino acids to alanine resulted in markedly reducing association of PTEN with NHERF-1. In addition, the full-length of PTEN robustly associated with NHERF-1 was also determined by co-immunoprecipitation experiment in cos-7 cells. Conclusion PTEN/NHERF-1 association was mediated via the binding of PTEN carboxyl-terminal]with the PDZ1 of NHERF-1, and the last four amino acids of the PTEN carboxyl-terminal were important for PTEN/NHERF-1 interaction.

OBJECTIVETo study the immuno-tolerance mechanism of the third-party bone marrow-derived mesenchymal stem cells (BMSCs) in the allogeneic transplantation.

METHODSForty female C57BL/6 mice and forty male BALB/C mice were respectively used as donors and recipients in skin allogenic graft model. Forty male BALB/C mice were divided randomly into 4 groups: blank control group, CP group, BMSCs group , CP + BMSCs group, with 10 mice in each group. Before skin graft, high-dose abdominal injection of cyclophosphamide (200 mg/kg, 2 d, q. d.) was performed in recipient mice in CP and CP + BMSCs groups. On the transplantation day, a bonus of 2 x 10(6) BMSCs from the SD rat (SD-BMSCs) were injected through the tail vein in the BMSCs and CP + BMSCs groups. The observation and HE staining of skin grafts were used. The expressions of CD29, CD34, CD45 and CD90 of cells were analyzed by using flow cytometry in order to identify BMSCs. The CD4+, CD25+, Foxp3 and Treg cells of spleen were detected by flow cytometry. Cytokine in peripheral blood of recipient mice were measured by ELISA, including TGF-beta, IL-10 and IFN-gamma. T cells were co-cultured with 60Co-irradiated bone marrow MSCs from different individuals. The proliferative activity of T cells were evaluated with MTT assay.

RESULTSThe skin graft survival time was significantly prolonged in the CP + BMSCs group, as compared with that in the blank control group, the CP group, the BMSCs group, respectively. Cells cultured by whole bone marrow adherent cultivation showed CD29 (99.7%), CD44+ (96.7%), CD34- (1.6%), CD45- (1.3%). Compared with the control group and CP group, the ratio of the CD4+, CD25+, Foxp3+ and Treg cells significantly increased in the SD-BMSCs group and CP + BMSCs group (P < 0.05). Analysis of peripheral blood by ELISA showed significant high level of TGF-beta, IL-10 and low level of IFN-gamma in BMSCs group and CP group,compared with that in control group. When co-cultured with BMSCs from different individuals, T- lymphocytes proliferation decreased apparently in SD-BMSCs group and C57-BMSCs group (P < 0.05), but there was no significant difference between SD-BMSCs group and C57-BMSCs group (P > 0.05).

CONCLUSIONSThe immunotolerance mechanism of the third-party bone marrow-derived mesenchymal stem cells in the allogeneic transplantation might be associated with its effect on the proliferation of Treg cells and increasing expression of TGF-beta and IL-10, decreasing expression of IFN-gamma.

Animals ; Bone Marrow Cells ; immunology ; Female ; Immune Tolerance ; Interferon-gamma ; immunology ; Interleukin-10 ; immunology ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; T-Lymphocytes, Regulatory ; immunology ; Transforming Growth Factor beta ; immunology ; Transplantation, Homologous

The aim of this paper is to apply near infrared spectroscopy techniques to construct a rapid identification method for 8 kinds of mineral Chinese Medicines containing carbonates. The qualitative model using clustering analysis method in OPUS software can identify accurately 8 kinds of carbonate-containing mineral Chinese medicines. The near-infrared quantitative model was established by using partial least squares method (PLS) for 7 mineral Chinese Medicines in which main component is calcium carbonate. Compared with the results by EDTA titration, the established quantitative analysis model for calcium carbonate content showed a good prediction result that when the content is between 47.61% -99.17%, the average relative deviation of the prediction result is 0.24% and the average recovery rate was 100.3%. The results also showed that the model using near infrared spectroscopy can get not only a rapid identification of the 8 mineral Chinese medicines containing carbonates, but also an accurate and reliabe content determination of calcium carbonate for the 7 mineral Chinese medicines which contain the component.
Carbonates ; analysis ; Medicine, Chinese Traditional ; Minerals ; chemistry ; Software ; Spectrophotometry, Infrared ; methods ; Time Factors

Objective To investigate the molecular types of carbapenem-non-susceptible Esche-richia coli ( E.coli) isolates and their mechanism of carbapenem resistance .Methods Twenty-two carbap-enem-non-susceptible E.coli strains were isolated from 3 hospitals in Hangzhou from 2007 to 2011.The mini-mum inhibitory concentrations ( MICs) of antimicrobials to those isolates were determined by agar dilution method and E-test.The molecular mechanisms of carbapenem resistance of E.coli isolates were analyzed by conjugation experiment,PCR and DNA sequencing.Pulsed-field gel electrophoresis (PFGE),multilocus se-quence typing ( MLST ) , and phylogenetic typing were performed to analyze the molecular epidemiology of those isolates.Results The MICs of imipenem and meropenem to 22 E.coli isolates were ranged from 1 μg/ml to 16 μg/ml,and the MICs of ertapenem were 2 μg/ml to 64 μg/ml.All E.coli isolates produced the KPC-2 carbapenemase and various β-lactamases , and some of them also produced plasmid-mediated AmpC enzymes.Carbapenem resistance was transferred by conjugation and transformation from 22 E.coli iso-lates to E.coli EC600 strains.The E.coli transconjugants or transformants acquired the blaKPC-2 gene and showed similar antibiotic susceptibility patterns in comparison with donor strains .Only a few isolates were in-distinguishable or closely related as indicated by PFGE .Four sequence types including ST131 (9 isolates), ST648 (5 isolates),ST38 (2 isolates) and ST405 (2 isolates) were identified by MLST.Phylogenetic analy-sis indicated that 9 ST131 isolates belonged to phylogenetic group B 2 and the other isolates belonged to group D (11 isolates),group B1 (1 isolate) and group A (1 isolate),respectively.Conclusion The sequence type of prevalent E.coli isolates producing KPC-2 from Hangzhou was ST131,which is an international epi-demic,multidrug-resistant clone,followed by ST648.