Objective To determine,in our experience,the prevalence and presence of risk factors for metabolic syndrome (MS) in heart transplant recipients.Methods We studied 135 patients who underwent heart transplantation in our hospital from September 2000 to December 2011,According to the diagnostic criteria of MS,they were divided into two groups.All variables that could be related to the development of MS during the follow-up period were analyzed.The prevalence and presence of risk factors for MS in our experience were determined.Student t-test,Wilcoxon rank sum test and x2 test were used for univariate statistical analysis and logistic regression for multivariate analysis.Results (1) Of the 135 patients,63 developed MS (46.67％) during a mean follow-up period of 47.3 months.The multivariate analysis identified the follows as predictive factors for the development of MS:age (OR=1.09,95％ CI 1.01-1.22,P＜0.05),body mass index (BMI) (OR=1.65,95％ CI 1.16-1.87,P＜0.05),history of DM (OR =3.27,95％ CI 1.12-34.21,P＜0.05).Conclusion In our population,the prevalence of MS after heart transplant is 46.67％ during a mean follow-up period of 47.3 months.Age,pre-operative overweight especially BMI,and history of DM were significant and independent risk factors for the development of MS during the follow-up period.For these patients,dose of immunosuppressant and diet control are important.

Objective To investigate the function of phosphatidylinositol phospholipase C encoded by LB361 gene of L.interrogans ( L-PI-PLC) and its mechanism in inducing macrophage apoptosis .Meth-ods The PI-PLC domains in the sequence of LB 361 gene of L.interrogans serovar Lai strain were analyzed by bioinformatics method .Prokaryotic expression system was established to express the recombinant L -PI-PLC ( rL-PI -PLC).The enzymatic activity of rL-PI-PLC in hydrolyzing phosphatidylinositol -4,5-bisphos -phate (PIP2) substrate into inositol-1,4,5-trisphosphate (IP3) was determined by IP3 fluorescence polariza-tion assay.LB361gene expressions at mRNA and protein levels as well as the secretion of LB 361gene prod -ucts were detected by real-time fluorescent quantitative RT -PCR and Western blot assay after infection of hu-man THP-1 macrophages with L.interrogans serovar Lai strain.A LB361 gene-transfected THP -1cell line was generated for evaluation of the mechanism of LB 361 gene products in elevating intracellular free Ca 2+( [Ca 2+] i) concentration and inducing the apoptosis of transfected THP -1 cells with the use of laser confocal microscopy and flow cytometry.Re sul ts The rL-PI-PLC hydrolyzed PIP2 into IP3 with a Km of 199 μmol/L and a Kcat of 8.566×10-5 S-1 .The expressions of LB361gene at mRNA and protein levels were both signifi -cantly up-regulated after infection of THP-1 cells with L.interrogans serovar Lai strain .Moreover , the exter-nal secretion of L-PI-PLC was also found during infection .The concentrations of IP 3 and [ Ca2+] i in the LB361 gene-transfected THP-1 cells were significantly increased compared to those in the non-transfected THP-1 cells, resulting in a high [Ca2+]i-dependent apoptosis of partial THP-1 cells.Conclusion PI-PLC is encoded by the LB361 gene of L.interrogans, which could induce the apoptosis of macrophages through el-evating [ Ca2+] i concentration during infection of microphages with L.interrogans.

ObjectiveTo detect the mRNA and protein expressions of Bmi-1 in hepatocellular carcinoma (HCC) tissue,pericarcinomatous tissue,portal vein tumor thrombus and normal liver tissue,and to investiage the significance of Bmi-1 in the genesis and progression of HCC.MethodsForty tissues of HCC were collected from the Tongji Hospital of Huazhong University of Science and Technology from January 2005 to December 2009.The mRNA and protein expressions of Bmi-1 in the HCC tissues (40 cases),pericarcinomatous tissues (40 cases),portal vein tumor thrombus ( 11 cases) and normal liver tissues ( 10 cases) were detected by immunohistochemistry,Western blot and real-time polymerase chain reaction.The relationship between the expressions of Bmi-1 and the clinicopathological factors was analyzed.Differences in each group were compared by using the Nemenyi test or Dunnet t test,and the relationship between the clinicopathological factors and the protein expression in the HCC tissues was analyzed by the chi-square test or Fisher exact probability.The survival curve was drawn by the Kaplan-Meier method,and the survival rate was analyzed by the Log-rank test.Results The median relative mRNA expressions of Bmi-1 in the normal liver tissues,pericarcinomatous tissues,HCC tissues and portal vein tumor thrombus were 0.96,2.60,7.51 and 29.95,respectively.The results of immunohistochemistry showed that the high protein expression rates of Bmi-1 in the normal liver tissues,pericarcinomatous tissues,HCC tissues and portal vein tumor thrombus were 10.0％,20.0％,67.5％ and 100.0％,respectively.The high protein expression rates of Bmi-1 in the HCC tissues and portal vein tumor thrombus were significantly higher than those in the normal liver tissues and pericarcinomatous tissues (x2 =17.25,22.77;22.04,23.95,P ＜ 0.05 ). High protein expression of Bmi-1 was also detected in 11 cases of HCC tissues with portal vein tumor thrombus.The results of western blot were consistent with those of the immunohistochemistry.The mRNA and protein expressions of Bmi-1 were correlated with Edmondson grade and portal vein metastasis ( x2 =5.572,P ＜ 0.05 ),whereas they were irrelevant to the tumor size,serum levels of α-fetoprotein and hepatitis B surface antigen ( x2 =0.000,0.019,0.663,P ＞0.05).Patients with high expression of Bmi-1 had poor prognosis.ConclusionsBmi-1 is correlated with the genesis and progression of HCC as well as the formation of portal vein tumor thrombosis.Patients with high Bmi-1 expression have poorer prognosis when compared with those with low Bmi-1 expression.

Objective To clone mcp1, mcp2 and mop3 genes that encoding methyl-accepting chemotaxis proteins(MCP) of Campy/obacter jejuni for construction of their prokaryotic expression systems, and to establish chemotactic model in vitro of the microbe for determining chemotaxis-inducing substances and to understand relationship among MCPs and chemotactic inducers. Methods The segments of mep1, mcp2 and mcp3 genes were amplified by PCR and then sequenced after T-A cloning. Prokaryofic expression systems of the genes were subsequently constructed. SDS-PAGE pins Bin-Rod Gel Image Analyzer were used to examine the expression of target recombinant proteins rMCP1, rMCP2 and rMCP3, and Ni-NTA affinity chromatography was performed to purify the rMCPs. Rabbits were immunized with each of the three rMCPs to obtain antisera. Immunodiffusion assay was performed to measure the titers of antisera. IgG in each of the antisera were extracted by ammonium sulfate precipitation and DEAE-32 ion exchange chromatography, and IgG F(ab')2 were then prepared by pepsin enzymolysis and Sephadex G-100 chromatography. Chemotactic model in vitro of C. jejuni based on HAP( hard-agar plus) method was established to determine the chemo-taxis-inducing effect of eight candidate substances. Chemotaxis inhibition test based on IgG F(ab')2 bloc-king was applied to determine the function and diversity of MCPs. Results The segments with expected si-zes amplified from mcp1, racp2 and mcp3 genes were obtained by PCRs, and their nucleotide and putative amino acid sequences were completely same as the reported. The constructed prokaryotic systems could effi-ciently express rMCPs with the yields about 10% of the total bacterial proteins. Immunization with rMCP1, MCP2 and rMCP3 enables the rabbits to produce specific antibody. All the antisera had 1: 4 immunodiffusion titers. Both bovine bile and sodium deoxycholate(DOC) were able to induce ehemotactie movement of C. je-juni in a dosage-dependent manner ( P < 0.05 ). When MCP1 and MCP2 were blocked with their IgG F(ab')2, the ehemotaetic ability of C. jejurd were remarkably decreased (P <0. 05). However, MCP3 blocking did not affect the chemotaxis ( P > 0.05 ). Conclusion The C. jejuni MCPs are successfully ex-pressed in this study. Bovine bile and DOC are the inducers for chemotaxis of C. jejuni, and MCP1 and MCP2 are involved in the process of ehemotaxis iadueed by DOC.

BACKGROUND:The need for bone graft and its substitutes has dramatically increased during the past decade. Either bio-derived graft materials or synthetic materials cannot satisfy this need. Study of composite bone graft has become a focused field.OBJECTIVE: To develop a novel porous poly (lactic-acid) (PLA)/bone matrix gelatin (BMG) bioactive composite material by means of supercritical carbon dioxide fluid technique (SC-CO2) and evaluate the bone forming potential in vitro.DESIGN: A comparative study and observation.SETTING: Research Center of Tissue Engineering, Southern Medical University; Institute of Biomaterials and Pharmaceutical Technique, China Institute for Radiation Protection.MATERTALS: Mouse osteoblast-like MC3T3-E1 cells were obtained from RIKEN Cell Bank in Japan. PLA was supplied by the Institute of Bio-technique of Jinan University. Alkaline phosphatase (ALP) kit was the product of Nanjing Jiancheng Bio-engineering Institute. MC3T3-E1 cells were cultured in Dulbecco's modified eagle medium (DMEM) (Gibco Laboratories, USA) supplemented with 10% fetal bovine serum (FBS, Si Ji Qing, China), 100 mg/L penicillin and 100 U/L streptomycin in a 37℃ humidified atmosphere. The cells were passaged with 0.25% trypsin (Gibco Laboratories, USA).METHODS: The experiment was carried out in the Research Center of Tissue Engineering, Southern Medical University from October 2005 to July 2006. The porous PLA/BMG composite biomaterials and PLA were prepared with SC-CO2 technique, and then macroscopic and microscopic observations were performed. The MC3T3-E1 cells were co-cultured with PLA and PLA/BMG in vitro, and DMEM was taken as the blank control group. Each well was pictured with digital camera.The percentage of the stained area, standing for the calcification deposition, was determined with an image processing and analysis software. The ALP activity and calcium content were determined with the method of ultrasonic cell lysis.MArN OUTCOME MEASURES: ① Macroscopic and microscopic observation of PLA/BMG; ③ Quantitative measurement of the calcification area; ② ALP activity and calcium content.RESULTS: ① Macroscopic and microscopic observation of PLA/BMG: The PLA/BMG showed good homological porosity with the size of 50-150 μm and connectivity: There were many holes with the size of 5-10 μm in the PLA/BMG walls. The PLA and BMG were mixed uniformly. ② Calcification areas: The percents of calcification area were significantly different among the PLA/BMG group, PLA group and blank control group [(42.98±4.44)%, (9.55±1.94)%, (0.86±0.41)%, P ＜ 0.01]. ③ Results of calcium content and ALP activity: The ALP activities were significantly different among the PLA/BMG group, PLA group and blank control group [(5 427.58±1173.57), (1 060.54±500.27),(40.01±24.50) nkat/g, P ＜ 0.05-0.01]; The calcium content in the PLA/BMG group was higher than those in the PLA group and blank control group [(3.51±1.64), (1.04±0.21), 0.70±0.24] mmol/g, P ＜ 0.01].CONCLUSTON: The PLA/BMG prepared by means of SC-CO2 has a good osteoinductive activity and it is worth studying further as bone biomaterial and bone tissue engineered scaffold.

OBJECTIVETo clarify the role of waist circumference (WC) in estimating organ damage and assessing the prognosis in young and middle-aged patients with cardiovascular disease undergoing cardiac rehabilitation program (CRP).

METHODSWe recruited 147 young and middle-aged patients with cardiovascular disease to participate in the CRP for 4 or 5 weeks. The baseline clinical data and results of routine fasting blood test and ultrasonic cardiography were evaluated for these patients.

RESULTSWC had a linear correlation with the indices reflecting cardiac structure alteration (LVSd and LVPWd) of the patients after CRP but showed a negative correlation with their HDL-C levels. The fasting blood glucose (FBG) and serum uric acid (UA) levels showed a positive correlation with WC before CRP but not after CRP.

CONCLUSIONWC monitoring allows accurate assessment of organ damage in patients undergoing CRP after cardiovascular disease and has special value in evaluating the changes in cardiac structure and function.

Blood Glucose ; chemistry ; Body Mass Index ; Cardiovascular Diseases ; diagnosis ; physiopathology ; Humans ; Middle Aged ; Prognosis ; Uric Acid ; blood ; Waist Circumference

Background: Resistin level is high in patients with severe acute pancreatitis (SAP), and resistin is expected to be a new marker for evaluating the severity of acute pancreatitis. Objective: To explore the influence of integrated traditional Chinese and Western medicine therapy on serum resistin levels in SAP patients. Design, setting, participants and interventions: Twenty-eight SAP patients meeting inclusion criteria from Department of Integrated Traditional Chinese and Western Medicine, West China Hospital, Sichuan University were included, and the patients were randomly divided into treatment group and placebo group. There were 13 patients in the treatment group and 15 patients in the placebo group. Patients in the treatment group were given traditional Chinese herbal medicine in addition to the conventional treatment. Patients in the placebo group were given placebo in addition to the conventional treatment. Main outcome measures: The serum resistin levels on admission, and days 1, 3, 5, and 7 after the admission were detected. Results: The serum resistin levels on admission in all the patients were higher than normal level, and there was no significant difference between the two groups (P>0.05). On days 1, 3, 5, and 7 after admission, the resistin levels in the treatment group were (3.29+/-1.66) mu g/L, (3.71+/-1.05) mu g/L, (3.08+/-1.47) mu g/L and (3.62+/-1.67) mu g/L, and in the control group (5.16+/-1.93) mu g/L, (5.07+/-1.53) mu g/L, (4.88+/-1.47) mu g/L and (5.12+/-1.48) mu g/L, respectively. The resistin levels were lower in the treatment group than in the control group (P<0.05). Conclusion: Serum resistin level in SAP patients can be decreased by integrated traditional Chinese medicine and Western medicine therapy.

Objective To investigate the relationship of serum resistin and pancreatic necrosis in the patients with severe acute pancreatitis. Methods Twenty-eight patients with SAP admitted to our hospital from March 2008 to November 2008 were divided into two groups according to the CT scan imaging: necrotic group and non-necrotic group. The enzyme-linked immunosorbent assay (ELISA) was used to test the serum resistin levels. An ROC curve was depicted to predict the pancreas necrosis. Results There were 21 patients in the non-necrotic group and 7 in the necrotic group, and there was no significant difference in terms of sex, age and baseline disease (P>0.05). The resitin levels ranged from 0.1730 ng/ml to 7.4923 ng/ml, with a mean (3.7102±1.6987) ng/ml. The area under the curve of resistin values was 0.884±0.108 (95%CI:0.672～1.097), asymptote signals 0.003, then it was calculated that P=0.003, which was>0.50. Conclusions The serum resistin may be of clinical value to predict the pancreatic necrosis.

Objective To evaluate the expression of transient receptor potential melastatin 7(TRPM7)in cholangiocarcinoma and its correlation with prognosis.Methods The expressions of TRPM7 were detected by SP immunohistochemical in 49 cases of cholangiocarcinoma,7 cases of benign bile duct lesions and 36 cases of adjacent histologically noncancerous bile duct tissues,and to analysis its relationship with the clinical pathological characteristics of cholangiocarcinoma.Results The positive expression rate of TRPM7 in cholangiocarcinoma was 77.6%(38/49),which was higher than that in benign bile duct lesions(0,0/7)and adjacent his-tologically noncancerous bile duct tissues(2.8%,1/36),the difference was statistically significant(P <0.05).The positive expres-sion of TRPM7 in cholangiocarcinoma was correlated with the TNM stage of tumor,lymph node metastasis and organ metastasis (P <0.05),but not related to patients′age,gender,site,differentiation and hepatitis(P >0.05).Kaplan-Meier analysis showed that increased expression of TRPM7 was associated with shorted overall survival (P <0.05).Cox regression analysis showed that the expression level of TRPM7 was significantly associated with prognosis and an independent risk factor for prognosis(P <0.05 ). Conclusion TRPM7 plays an important role in the tumorigenesis progression,invasion,and metastasis of cholangiocarcinoma,and is an important factor for prognosis in patients with cholangiocarcinoma.

Objective To investigate the levels of oxidative stress in liver tissues of hepatocelluar carcinoma(HCC)patients after transcatheter arterial chemotherapy(TAC).Methods Immunohistochemistry streptavidin biotinylated peroxidase(S-P)method was used to detect the cellular levels of 8-hydroxy-2'-deoxyguanosine(8-OHdG),p53 and p21~(waf1/cip1).Eighty-nine HCC patients were divided into TAC group(39 cases)and Non-TAC group(50 cases).15 Non-HCC liver tissues served as controls.Result 8-OHdG level was higher in Non-TAC group than that in TAC group in tumor tissues (F=9.516,P＜0.05),with that being the lowest in control group(F=9.516,P＜0.01);8-OHdG levels in cancer tissues were significantly higher than that in tumor surrounding tissues in both TAC group (t=7.101,P＜0.001)and Non-TAC group(t=8.020,P＜0.001),there was no significant difference of 8-OHdG levels between para-tumor tissues and controls.The levels of 8-OHdG between tumor and its surrounding tissues in TAC group(r=0.651,P＜0.001)and non-TAC group(r=0.493,P＜0.01)was in positive correlation.The difference of p53 levels in cancer tissues in TAC group and Non-TAC group were not statistically significant and p53 was not detected in para-tumor tissues.The difference of p21~(waf1/cip1) levels among TAC group,Non-TAC group and controls was statistically significant,the levels of p21~(waf1/cip1) in normal group was the highest(F=13.459,P＜0.001),followed by that in TAC and Non-TAC group in cancer tissues(TAC vs.Non-TAC group,P＜0.01);p21~(waf1/cip1) expression in normal controls was significantly higher than that in both TAC and Non-TAC group in para-tumor tissues(F=16.613,P＜0.001).The correlation of p21 ~(waf1/cip1) levels between tumor and its surrounding tissues was significant in non-TAC group(r=0.872,P＜0.001).Conclusions Oxidative stress levels in HCC tumor tissues were higher than in para-tumor tissues and non-HCC liver tissues.Cancer cells probably survive chemotherapy by fortifying oxidative stress repair mechanism.