Seven genes (fabD, fabG, fabH, fabA, fabZ, fabB and fabI) of E.coli fatty acid biosynthetic enzymes were cloned by PCR amplifying and appropriate expression vectors were constructed. Under induction assay expression of plasmid encoded proteins was carried out in strain BL21(DE3) and seven enzymes were purified using Ni-NTA agarose resin. In the absence of [2-14C] malonyl-CoA fatty acid synthetic reaction was reconstituted in vitro by adding seven enzymes and co-factors. And several model reactions were established for identification of special fatty acid biosynthetic enzymes. Meanwhile Clostridium acetobutylicium FabZ function was characterized by this method.

Objective The purification methods of the exosomes derived form T cells were established in order to get high quan-tity exosomes .Methods Exosomes from T cells culture supernatants were purified by ExoQuick Precipitation ,ultrafiltration and sucrose gradient centrifugation ,differential ultracentrifugation ,and confirmed via using transmission electron microscopy .The pro-tein expression of the exosomes were analyzed by SDS-PAGE electrophoresis .Western blotting was used to test the expression of IL-2 .Results The protein concentration of the exosomes purified through ExoQuick Precipitation ,ultrafiltration and sucrose gradi-ent centrifugation were higher than through differential ultracentrifugation (P<0 .05) .SDS-PAGE displayed the difference among the exosome purified by three methods .Three kinds of exosomes all expressed IL-2 .Conclusion ExoQuick Precipitation ,ultrafiltra-tion and sucrose gradient centrifugation technique can obtain high purity and complete exosome sample .

FabB(?-ketoacyl-acyl carrier protein synthase Ⅰ)and FabF(?-ketoacyl-acyl carrier protein synthase Ⅱ)are two key enzymes of fatty acid biosynthesis in E.coli.The Gram-positive pathogenic bacterium Enterococcus faecalis has a fatty acid composition very similar to that of E.coli.Bioinformatic analysis reveals that though E.faecalis has two fabF homologues,there is no recognizable fabB homologue in the genome of E.faecalis.Two fabF homologues(fabF1 and fabF2)were amplified by using E.faecalis V583 genomitic DNA as template,and two plasmids,pHW13(fabF1)and pHW14(fabF2),were constructed.The results of experiments in vivo and in vitro have shown that fabF1 gene could complement E.coli fabB mutation and FabF1 possessed ?-ketoacyl-acyl carrier protein synthase Ⅰ(FabB)activity,while fabF2 gene could complement E.coli fabF mutation and FabF2 had ?-ketoacyl-acyl carrier protein synthase Ⅱ(FabF)activity.Meanwhile the data also shown that FabF2 possessed partial function of ?-ketoacyl-acyl carrier protein synthase Ⅰ(FabB),and it could make E.coli fabB mutation synthesized low amount of unsaturated fatty acid.From these data it is clear that FabF species enzymes could have activity of ?-ketoacyl-acyl carrier protein synthase Ⅰ(FabB).

Objective To study the effect of Tacrolimus on blood lipid after renal transplantation,and the relationship between C825T polymorphism in G protein beta 3 subunit (GNB3) gene and serum lipid levels.Method Eighty-one cases of recipients patients after renal transplantation were divided into two groups in terms of Tacrolimus concentration:normal blood concentration group (group A) and low blood concentration group (group B).The serum lipid levels at 1st,3rd,6th,and 12th month after renal transplantation were measured.Genotype was determined by the simple sequence-specific primer polymerase chain reaction (SSP-PCR).Result The percentage of patients with hypertriglyceride in group A was significantly higher than in group B during the one-year follow-up period.There was significant difference between the two groups in the serum triglyceride levels but no difference in the serum cholesterol levels.The 825C/T polymorphism in the GNB3 gene was not associated with hypertriglyceride in renal transplantations in Wenzhou.Conclusion The serum triglyceride levels in renal transplantations in Wenzhou was associated with the Tacrolimus concentration,and the incidence of hypertriglyceride is not associated with the 825C/T polymorphism in the GNB3 gene.

Objective To investigate the protein expression and genetic alterations of c-myc in primary systemic anaplastic large cell lymphoma (ALCL) and discuss its relationship with clinicopathologic features and immunophenotypes.Methods 87 cases of ALCL were selected.Immunohistochemical method was used to detect the protein expression of c-myc,ALK,CD3,CD10,CD20,CD30 and EMA.c-myc and ALK genetic alterations were detected by using fluorescence in situ hybridization (FlSH).The interrelationships between protein expression,genetic alterations and clinicopathological parameters were analysed statistically.Results Immunohistochemical results:of 87 cases,ALK protein was expressed in 54 cases (62.1％).c-myc protein was expressed in 27 cases (31.0 ％).ALK and c-myc were co-expressed in 20 cases (23.0 ％).c-myc protein expression,ALK and c-myc co-expression increased with the upgrade of ALCL clinical stages,and the expression was higher in International Prognostic Index (IPI) high-risk groups than in low-risk groups (P ＜ 0.05).FISH test results:of 87 ALCL cases,there were 50 cases (57.5 ％) of ALK rearrangements and 19 cases (21.8 ％) of ALK aneuploidy.c-myc rearrangement was detected in none of 87 ALCL cases,but there was aneuploidy in 19 cases (21.8 ％).The differences of c-myc aneuploidy in ALK positive and negative groups were statistically insignificant (P ＞ 0.05),while they were statistically significant in c-myc groups (P ＜ 0.05) and in different IPI groups (P ＜ 0.05).Conclusion c-myc protein expression and aneuploidy were related with ALCL clinical stages and IPI,which could be used as an indicator of estimating ALCL malignant degree and predicting prognosis.

Objective To investigate the dysregulation of HER-2 protein and gene in non-small cell lung cancer (NSCLC), and to identify the association between clinicopathological features,prognosis and HER-2 aberrations amongst protein and gene. Methods 140 NSCLC tissues (89 squamous cell carcinoma, 51 adenocarcinoma) with operative section and detailed case were taken from pathology department of Shanxi Cancer Hospital from Jan 2006 to Feb 2007, while 70 normal tissues were set as control group. Immunohistochemistry was applied to detect the state of HER-2 protein expression,and fluorescence in situ hybridization (FISH) was applied to test the status of gene amplification. Results In normal and NSCLC tissues, over-expression of HER-2 was detected in 0 case and 17 (12.14 %) cases (P < 0.05), respectively. The over-expression of HER-2 was associated with the pathological type of NSCLC, which was detected more frequently in adenocarcinoma (χ2 = 4.19, P = 0.04), rather than the gender, age, smoke history, clinical stages, and lymphatic metastasis of patients. 40 (28.57 %) cases presented HER-2 gene copy number ≥3, including 6 (4.29 %) patients with HER-2 gene amplification, 34 (24.29 %) patients with HER-2 gene multicopy. HER-2 gene amplification was associated with the pathological type (P = 0.024), smoke history (P = 0.048) and age (P = 0.015), rather than lymphatic, gender, clinical stages. None clinicopathological features were presented correlation with HER-2 gene multicopy (P > 0.05). There was no significantly difference in survival between patients with and without HER-2 protein over-expression and HER-2 gene dysregulation (P > 0.05). HER-2 protein over-expression was associated with HER-2 gene amplification (P > 0.05), while no relationship between HER-2 protein overexpression and HER-2 gene multicopy (P < 0.01). Conclusions The over-expression of HER-2 is related to pathological type of NSCLC with more frequent expression in adenocarcinoma. The incidence rate of HER-2 gene amplification in patients with adenocarcinoma histology, never-smokers, and young age is high. The HER-2 protein over-expression and gene dysregulation show no relation with the prognosis of NSCLC.

Objective To analyze factors of volume maintenance percentage after autologous fat grafting breast augmentation.Methods From March 2011 to December 2014,39 patients were included.Age,BMI,donor site,breast preoperative volume,whether or not application of Brava and water-jet-assisted liposuction were collected as variables.We performed multivariate regression and univariate regression analyses in a general linear regression model and multilevel linear regression model to identify significant factors.Results The mean percentage of volume maintenance was (34± 12) ％.In univariate regression analysis,rate of volume maintenace was decreased 3.3％ as increase of age per 5 years;Brava application increased volume maintenace,compared with donor sites of the back and abdomen 8.4％ (40.7％ vs 32.3％,P＜0.05),application of leg fat increased by 9.6％ of volume maintenace (39.1％ vs 29.5％,P＜0.05).In the multivariate regression analysis,Brava use increased volume maintenance of 6.2％ (P＜0.05);use of leg fat increased by 6.0 ％ of volume maintenace as compared with donor sites of the back and abdomen (P＜0.05).Conclusions The age,Brava,and donor site are significant impact factors to percentage volume maintenance in autologous fat graft breast augmentation.

[ABSTRACT]AIM:ToinvestigatetheeffectofimmunosuppressantFK506onserumglucoseinratsandtoex-plore its mechanism .METHODS: Sprague-Dawley rats ( n =12 ) were randomly divided into drug group and normal group.The rats in drug group were intraperitoneally injected with FK 506 at dose of 1 mg· kg-1 · d-1 and the rats in nor-mal group received saline (1 mL· kg-1 · d-1 , ip) for 14 d.The fasting weight and fasting glucose were regularly meas-ured every 2 d.Visceral fat was isolated from the rats at the end of experiment .The mRNA expression of adiponectin , lep-tin, visfatin, resistin, retinol-binding protein 4 ( RBP4) and peroxisome proliferator-activated receptors γ( PPAR-γ) was determined by real-time fluorescence quantitative PCR .The protein expression of PPAR-γand adiponectin was measured by Western blotting .RESULTS:Compared with normal group , the concentration of fasting blood glucose in model group was significantly increased from the 10th day (P<0.05).At day 14, the fasting blood glucose of the model group increased from (5.10 ±0.62) mmol/L to (7.73 ±0.73) mmol/L.No significant change of blood glucose in normal group between the 10th day and the 14th day [from (4.66 ±0.32) mmol/L to (5.80 ±0.10) mmol/L] was observed.Compared with normal group , the mRNA expression of PPAR-γ, adiponectin and leptin in the adipose tissue of model group was signifi-cantly decreased ( P <0.01 ) , whereas the expression of visfatin , resistin and RBP4 was significantly increased ( P <0.05).Compared with normal group, the expression of PPAR-γand adiponectin in model group was decreased (P <0.01).CONCLUSION:FK506 may decrease the expression of PPAR-γto change the expression of adipocytokines and induce hyperglycemia in rats .

Objective To investigate the association of genetic polymorphisms in glutathione S-transferases T1 (GSTrl), M1 (GSTM1) and P1 (GSTP1) with aristolochic acid nephropathy (AAN) of Chinese people in Wenzhou of China. Methods Fifty-nine patientswith AAN (AAN group) including 29 male and 30 female as well as 157 healthy ethnically matched controls (control group) including 93 male and 64 female were enrolled in this study. The genotypes of GSTT1, GSTMI and GSTP1 were determined by multiple PCR and confronting two-pair primers PCR (CTPP-PCR). Results The genotype frequencies of GSTP1 were in Hardy-Weinberg equilibrium. Compared with the healthy controls, the frequency of GSTT1 null genotype was significantly higher in the patients with AAN (66.1% vs 48.4%,P＜0.05). Risk of A.AN for individuals with GSTT1 null genotype was 1.747 fold of those without GSTIl null genotype (95% CI=0.818-3.731). The frequency of GSTM1 null genotype, GSTP1 variant genotypes and GSTP1 G allele in the patients and in the controls were 40.7%, 28.8%, 16.1% and 47.8%, 31.8%, 17.5%, respectively, which were not significantly different. No significant differences were found in prevalence of GSTM1 and GSTP1 gene distribution between patients and controls. Conclusion GSTrl gene polymorphism appears to be associated with susceptibility to AAN in Southern China.

This article describes a novel Multifunctional and Transparent Urinary System Model (MTUSM), which can be applied to anatomy teaching, operational training of clinical skills as well as simulated experiments in vitro. This model covers kidneys, ureters, bladder, prostate, male and female urethra, bracket and pedestal, etc. Based on human anatomy structure and parameters, MTUSM consists of two transparent layers i. e. transparent organic glass external layer, which constraints the internal layer and maintains shape of the model, and transparent silica gel internal layer, which possesses perfect elasticity and deformability. It is obvious that this model is preferable in simulating the structure of human urinary system by applying hierarchical fabrication. Meanwhile, the transparent design, which makes the inner structure, internal operations and experiments visual, facilitates teaching instruction and understanding. With the advantages of simple making, high-findelity, unique structure and multiple functions, this model will have a broad application prospect and great practical value.
Female ; Humans ; Kidney ; Male ; Models, Anatomic ; Models, Biological ; Prostate ; Ureter ; Urethra ; Urinary Bladder ; Urogenital System ; anatomy & histology