Objective To study the distribution and molecular characteristics of Vibrio parahaemolyticus(VP) in patients with a‐cute diarrhea ,and provide a scientific basis for the prevention and control of VP infection .Methods From 2010 to 2014 ,62 VP iso‐lates were collected from patients with acute diarrhea ,for serotyping and virulence gene (tdh and tdh) detection of VP .Molecular characteristics analysis was carried out by using multi‐locus sequence typing (MLST) .Results 7 different serotypes were found from the 62 isolates .O3∶K6 was the most common serotype of VP ,accounting for 74 .19% (46 isolates) ,followed by O4∶K68 (6 isolates) .Tdh gene was the mainly virulence gene ,with a percentage of 95 .16% (59 isolates) ,only three isolates were trh positive . 7 STs were found through MLST analysis of 62 VP isolates ,among which ,ST3 was the most important type ,accounted for 85 .50%(53 isolates) .Conclusion O3∶ K6 serotype VP was the most prevalent type .Tdh gene is the most important virulence gene of WP .ST3 was the the dominant epidemic type .

AIM: To prepare a soluble hemangiopoietin(HAPO) protein and to construct pET22b(+) expression vector, to obtain pure recombinant HAPO protein and to measure its bioactivity. METHODS: HAPO cDNA was amplified using RT-PCR method from a commercial human fetal liver cDNA library. The resulting product was cloned into pET22b(+) vector and transformed into E.coli BL21(DE3). The recombinant protein was isolated and purified by Ni~(2+)-NTA chelating resin and the chromatographies of SP Sepharose FF. The adhesion of human umbilical vein endothelial cells (HUVEC) were measured by adhesion assay. RESULTS: HAPO gene with a reading frame of 897 bp was successfully cloned from human fetal liver cDNA library, the expressed pET22b(+)-HAPO fused protein existed in a soluble form, with the yield above 10% total bacterial protein and its purity achieved above 80%. The activity assay showed that the treatment of HAPO enhanced total adherence of HUVEC in a concentration-dependent manner. CONCLUSION: HAPO protein can be expressed in a soluble form. HAPO may facilitate the homing of hematopoietic stem/progenitor cells in vitro.

Objective To explore the characteristics of food preferences in children with autism spectrum disorder,in order to provide some reference for treating the odd diet habits.Methods Comparing food selection styles through a self-made parents questionnaire including 113 kinds of common food in six categories in China in 162 cases of neurotypical children (NC) and 162 cases of children with autism (AC) to investigate the food preference characteristics of autistic children.Results Compared with NC,AC had narrow food repertoire in grain (NC refuse 0.0(1.0),AC refuse 1.0(2.0),P＜0.001),beans(NC refuse 0.0(1.0),AC refuse 1.0(2.0),P＜ 0.05),meat (NC refuse 0.0 (0.0),AC refuse 0.0 (2.0),P＜ 0.05),vegetables (NC refuse 3.0 (5.0),AC refuse 6.0(1.0),P＜0.001) and fruits(NC refuse 0.0(1.0),AC refuse 2.0(5.0),P＜0.001).There was no difference in the food preferences in neurotypical children of different gender.However,AC boys were more selective in the grain (NA girls refuse 0.0 (1.0),AC boys refuse 1.0 (2.0),P＜ 0.05) and vegetables (NA girls refuse 3.5 (5.0),AC boys refuse 7.0(11.0),P＜0.05) than AC girls.Moreover,AC had higher preference to lower acceptance in most of food than NC,but not instant noodles(NC acceptance 71.01％,AC acceptance 81.02％,P＜0.05) and chilli (NC acceptance 20.71％,AC acceptance 28.47％,P＜0.05).Conclusion Compared with NC,AC have narrower food repertoire.On the other hand,AC have significantly higher acceptance of stimulating food like chili and instant noodle.

Objective To investigate the exercise performance in maintenance dialysis patients,and analyze its correlative factors.Methods Maintenance dialysis patients admitted in Tongji Hospital of Shanghai from December 2014 to March 2015 were enrolled,with their baseline data and biochemical measurement being collected.The anthropometric indexes including arm circumference,triceps skinfold,waist circumference and hip circumference were detected.The exercise activity was assessed by hand grip test,timed up and go test (3mTUG) and five times sit-to-stand test (FTSST).Patients were divided into fast group (3mTUG≤12 s) and slow group (3mTUG ＞ 12 s).Univariate and multivariable analyses were used to evaluate the factors influencing exercise performance in maintenance dialysis.Results There were 121 patients enrolled:62 on peritoneal dialysis and 59 on hemodialysis,76 men and 45 women.Patients' average age was (61.6± 13.0) years and median dialysis age was 31.7(12.3,69.0) months.There was no statistical difference between fast group (n=80) and slow group (n=41) in gender,dialysis method,dialysis age,body mass index (BMI),arm muscle area (AMA),waist-hip ratio (WHR),hemoglobin (Hb) and total cholesterol (TC).Patients in fast group were younger,had higher serum albumin,prealbumin,serum phosphate and iPTH,and less prevalence of diabetes than those in slow group.In exercise activity,patients in fast group had better performance in handgrip,3mTUG and FTSST (all P ＜ 0.05).Univariate analysis showed that,handgrip was correlated with sex (male),AMA,BMI,age,diabetes,serum phosphorus and TC;scores in FTSST was correlated with age,BMI,diabetes,WHR,dialysis method,dialysis age,prealbumin and serum phosphorus;scores in 3mTUG was correlated with age,diabetes,WHR,dialysis method and dialysis age,prealbumin,serum phosphorus and iPTH (all P ＜ 0.05).Multiple stepwise regression analysis showed that sex (male),age,AMA and diabetes were independently correlated with handgrip in dialysis patients (all P ＜ 0.05);age,dialysis method,BMI and diabetes were independently correlated with scores in FTSST (all P ＜ 0.05);age,dialysis method,diabetes and WHR were independently correlated with scores in 3mTUG (all P ＜ 0.05).Conclusions The exercise performances of patients on maintenance dialysis are impaired.Age and diabetes are independent factor associated with the exercise performances of patients on maintenance dialysis.AMA is independently associated with upper limb movement,and dialysis method,BMI and WHR are independent factors associated with lower limb movement in dialysis patients.

Objective:To investigate the effect of Xanthium mongolicum Kitag. ethanol extract (EXM) on airway inflammation induced by house dust mite (HDM) in mice with allergic asthma, and to explore the possible mechanism. Methods:The mice were randomly divided into blank group, model group, positive control group and low, medium and high dose groups, with 6 mice in each group. The low, medium and high dose groups were intragastrically administered with 100, 200 and 400 mg/kg of EXM, and the positive control group was given 1.56 mg/kg salbutamol solution. Mice in the blank group and the model group were intragastrically administered with equal volume of solvent, once a day for 8 consecutive days. Except the blank group, the other groups were sensitized with HDM on the 3rd, 4th and 5th days after administration to establish asthma models. After administration, the number of eosinophils (EOS), total cells and various kinds of cells (neutrophils, macrophages, lymphocytes and EOS) in bronchial irrigation fluid (BALF) of mice were observed. The levels of serum IgE and inflammatory factors (IL-4, IL-5, IL-10 and IL-13) in BALF and lung tissue were measured by enzyme-linked immunosorbent assay (ELISA). Pathological and inflammatory changes of bronchial smooth muscle in mice were observed by HE and PAS.Results:Compared with the model group, the number of EOS significantly decreased ( P<0.01 or P<0.05), the total cells count, macrophages, neutrophils, lymphocytes and EOS in BALF significantly decreased ( P<0.01), the content of IL-4 (142.59 ± 16.51 ng/L, 70.47 ± 20.63 ng/L vs. 212.18 ± 58.51 ng/L), IL-5 (57.49 ± 5.49 ng/L, 47.47 ± 6.30 ng/L vs. 72.65 ± 19.11 ng/L), IL-10 (98.51 ± 18.31 ng/L, 71.85 ± 9.15 ng/L vs. 120.16 ± 23.35 ng/L), IL-13 (85.81 ± 23.66 ng/L, 39.99 ± 17.37 ng/L vs. 149.07 ± 33.19 ng/L) in BALF significantly decreased ( P<0.05), and the content of IL-4 (154.55 ± 25.70 ng/L, 143.30 ± 22.19 ng/L vs. 214.68 ± 31.95 ng/L), IL-5 (87.49 ± 5.95 ng/L, 61.84 ± 8.14 ng/L vs. 97.65 ± 12.10 ng/L), IL-10 (91.28 ± 18.69 ng/L, 62.04 ± 9.39 ng/L vs. 117.80 ± 20.72 ng/L), IL-13 (196.48 ± 15.90 ng/L, 132.02 ± 6.86 ng/L vs. 238.42 ± 28.41 ng/L) in lung homogenate significantly decreased ( P<0.05), the serum IgE level in the high dose group significantly increased ( P<0.05). Conclusion:The Xanthium mongolicum Kitag. can inhibit allergic asthma induced by HDM, which may be through inhibiting the secretion of inflammatory cytokines by pulmonary macrophages.

Objective To analyze the correlation between the expression of Ki-67 and Her-2 and clinical features in breast invasive ductal carcinoma .Methods Clinical and pathological data of 202 female breast invasive ductal carcinoma patients were collected between January 2012 and September 2014 .The correlation between the expression of Ki-67 and Her-2 and clinical features was analyzed .Results The positive rate of Ki-67 was 73.3%(148/202) and that of Her-2 was 19.3(39/202).The expression of Ki-67 was related to histological grading , ER and PR status(P<0.05),but not to age, tumor size or lymph node status(P>0.05).The expression of Her-2 was related to ER and PR status (P<0.05), but not to age, tumor size, histological grading, or lymph node status(P>0.05).Moreover, the low expression Ki-67 and negative expression of Her-2 were positvely corelated with the positive expression of hormone receptor [estrogen receptor(ER),pro-gestin receptor(PR)](P<0.05).Conclusion High expression of Ki-67 suggested that the differential grade of tumor was lower , malignant grade was higher , infiltration was stronger and metastasis was easier .High expression of Ki-67 and positive Her-2 suggests lower sensitivity to endocrine therapy and treatment .Detection of Ki-67 and Her-2 expression in breast inva-sive ductal carcinoma is of guiding significance for understanding the degree of malignancy and for evaluating prognosis .

Objective To explore the effects of electroacupuncture on brain derived neurotrophic factor (BDNF) of rats with sciatic nerve injury (SNI); To discuss its biological mechanism for treatment of SNI. Methods Fifty adult male Wistar rats were chosen, and the sciatic nerves of rats were cut off and pulled on both sides of the cut ends into nerve regeneration chamber. The rats were randomly divided into normal group, sham-operation group, model group, and electroacupuncture group. In the electroacupuncture group, the rats were treated by electroacupuncture for 28 days. After the treatment, the nerve regeneration was observed through HE staining. Immunofluorescence was used to analyze the expression changes of BDNF in the nerve tissue and spinal cord. ELISA was used to observe the changes of expression of serum BDNF. Results The amount of axon regeneration in the electroacupuncture group was obviously more than that in the model group, and the outline of the tissue more clear. Electroacupuncture could promote the expression of BDNF in the nerve, spinal cord and serum of SNI of rats compared with model group (P<0.01). Conclusion Electroacupuncture can promote the repairment and regeneration of SNI in rats by upregulating the expression of BDNF.

Objective To evaluate the feasibility of real-time quantitative PCR (RtPCR) with small volume regarding the stability, efficiency and reliability of amplification, and determine the optimal quantity of cDNA template suitable for small PCR volume. Methods The experiment was carried out in 3 groups with 10, 15 and 20μL reaction volume, respectively. In each group, rat β-actin mRNA was detected by RtPCR with 0.1, 0.2, 0.3 or 0.4μL rat cDNA as template, respectively. The amplification curve and melting curve were used to evaluate the reaction stability. The fitting of a curve of gradient templates against threshold cycle numbers was to show the reaction efficiency and the linear correlativity was to estimate the suitability of the template quantity. In addition, in order to estimate reliability, pristane-induced arthritis (PIA) rat model was established, and spleen TNF-α mRNA expression was detected by RtPCR with the selected reaction volumes. Results The amplification of rat β-actin mRNA was specific and stable in 10μL, 15μL and 20μL PCR volume, and had a high efficiency. Furthermore, the standard curves fitted by 0.1-0.4μL gradient templates showed a significant linear correlation in each volume group. When the 10μL and 20μL PCR volumes, and 0.2μL cDNA templates were chosen, the TNF-α mRNA expression in PIA rat spleen showed significant upregulation in both two volume groups as anticipated. Conclusion The experiment shows that it is feasible in the RtPCR amplification to use the small reaction volume of 10μL and 15μL, which has good stability and reliability. And 0.1-0.4μL templates are all suitable for the reaction system. PCR with small volume can not only save the reagents and template, especially rare clinical specimens, but also is helpful for the realization of high-throughput reaction.

Objective To evaluate the influence of long-term nucleotide reverse transcriptase inhibitors (NRTIs) on lipids metabolism in HIV/AIDS patients and correlating clinical factors.Methods A total of 118 HIV/AIDS patients were divided into 3 groups:untreated group (40 patients),highly active antiretroviral therapy(HAART) for 1-2 years group (37 patients) and HAART over 5 years group (41 patients),with 20 healthy individuals as the control group.Clinical lipodystrophy (LD) was defined as concordance between patient's report of change and physical examination.Fat mass (FM) was measured by dual-energy X-ray absorptiometry (DXA).Results There was no significant difference in the incidence of LD between HAART for 1-2 years group and HAART over 5 years group (51.2％ vs 40.5％,P =0.345).The prevalence of LD was 2.4 folds with strvudine (d4T) treatment compared with zidovudine (AZT)-containing regimens (61.6％ vs 23.5％,P =0.001).Based on DXA measurements,FM of total body and limbs were significantly lower in the HAART over 5 years group than that in the control group,the untreated group and the HAART for 1-2 years group (P ＜ 0.05).Trunk FM was significantly lower in the HAART over 5 years group than the untreated group and the HAART for 1-2 years group (P ＜ 0.05).FM of total body and trunk were significantly lower in patients without LD in the HAART over 5 years group than patients without LD in the HAART for 1-2 years group (P ＜ 0.05).FM was correlated positively with body weight and BMI.Limbs FM was correlated negatively with peripheral blood triglyceride concentration.Conclusions HIV/AIDS patients with NRTIs therapy have high prevalence of LD,which mainly occurs 1-2 years after therapy,and increases with d4T treatment compared with AZT-containing regimens.There was no significant difference in the incidence of LD between the HAART for 1-2 years group and the HAART over 5 years group.FM was significantly decreased after long-term HAART in the patients with or without LD.DXA can evaluate LD objectively and guide further clinical treatment.

Objective To observe the effects of electroacupuncture on the expressions of autophagy related protein Beclin-1 and apoptosis related protein p53 of hippocampus in rats;To explore the mechanism of electroacupuncture on Alzheimer's disease (AD).Methods The rats were randomly divided into the normal group, the sham-operation group, the model group, and the electroacupuncture treatment group. “Baihui” and “Yongquan” points were taken for electroacupuncture treatment and the treatment course was 7 days. The rats were treated once a day for 4 courses. Changes in morphology and number of Nissl positive cells were examined by Nissl staining in hippocampal CA1 regions. Expressions of Beclin-1 and p53 protein were determined by Western blot analysis.Results Number of Nissl positive cells in CA1 region of the model group was significantly less than that of normal group (P<0.01). After electroacupuncture treatment, number of pyramidal cells and expression of Nissl body significantly increased (P<0.05). Expression of Beclin-1 decreased, while expression of p53 increased in the hippocampus of the model group, compared with that in the normal group (P<0.05). However, electroacupuncture treatment could significantly upregulate the expression of Beclin-1 protein (P<0.01), but downregulate the level of p53 (P<0.05).Conclusion Electro-acupuncture treatment could fight against Aβ-induced neuronal apoptosis, and improve the morphological changes of AD’s hippocampus.