Adult ; Anaphylaxis ; etiology ; Female ; Humans ; Male ; Semen ; immunology

Objective To screen and identify protein interacted with enterovirus 71 (EV71)3A protein by means of T7-phage display system.Methods The prokaryotic expression vector of 3A protein was constructed for expressing and purifying 3A pro-tein.With 3A protein as the target protein,the T7-phage display system was adopted to screen the human hepatocyte cDNA library. The screened products were performed the DNA sequence analysis and the homologous research.Results 3A protein was expressed and purified,after 4 rounds of biopanning,37 positive clones were selected.The inserted fragments were amplified by PCR using specific T7 primer.The products were sequenced,2 proteins were identified to interact with 3A protein by homologous analysis. Conclusion Trough the T7 phage display system,two proteins interacted with 3A are obtained.The possible function of 3A protein can be speculated by researching the function of these two proteins,which lays the foundation for the further studying the pathogen-ic mechanism of EV71.

Objective To explore the animal model making method for diarrhea-predominant irritable bowel syndrome (IBS-D) caused by the disharmony of liver and spleen in TCM.Methods In order to define the period of the animal model making,the 32 neonatal rats were divided into 3 days group,6 days group,9 days group and control group.In order to determine the weight of the experimental rats,the 48 rats with different weight were divided into lower weight model group,lower weight control group,medium weight model group,medium weight control group,higher weight model group and higher weight control group.In order to decide the method of causing diarrhea,the 32 rats were divided into lactose enriched diet group,restraint stress group,lactose enriched diet plus restraint stress group,and a control group.The body weight and diarrhea rate of all groups were compared respectively.After the method of model making was decided,the 16 rats were divided into a model group and a control group,and 5-hydroxytryptamin (5-HT),lactate dehydrogenase (LDH) and aquaporin 4 (AQP4) of the two groups were compared for model-proving.Results Based on the results step by step,the best method was decided:the period of the animal model making was 6 days,rats with higher body weight should be subjects,and the lactose enriched diet combining with restraint stress should be used to induce diarrhea.Model-proving test:compared with the control group,5-HT and LDH decreased (P

Child ; Cough ; chemically induced ; Disinfectants ; adverse effects ; Humans ; Irritants ; adverse effects ; Male

Objective: This study was undertaken to access the impact of hyperthermic chemotherapy (HTCT) on human gastric cancer cells in vitro. Methods: Six human gastric cancer cell lines (AGS, MKN45, SGC7901, NCI-N87, SNU-1 and SNU-16) and gastric cancer cells originated from clinical patients were used in this study. Treatment conditions were categorized into 4 modes: normothermic control (NT, 37°C), hyperthermia (HT, 43°C), normothermic chemotherapy (NTCT) and HTCT. The cells were treated with different concentrations of cis-diammine dichloroplatinum (CDDP, 0 to 32 mg/L) in combination with hyperthermia for 2 h. MTT assay was adopted to evaluate cell proliferation and cytotoxicity after HTCT. Preliminary morphological changes were observed under light microscope (LM) while ultrastructure changes and the definite cell death type were determined by transmission electron microscopy (TEM) and fluorescent microscopy (FM). The quantitative ratio of apoptosis and necrosis was determined by Annexin V-FITC and PI double staining using flow cytometry (FCM). Results: According to MTT assay and LM observation, there was a synergistic effect between hyperthermia and chemotherapy in inhibiting proliferation of the different types of gastric cancer cells. Hyperthermia enhanced the sensitivity of gastric cancer cells to CDDP chemotherapy. The concentration of CDDP required for inhibiting cell proliferation and inducing cell death was much lower in HTCT group than that of NTCT group. TEM and FCM examination showed that HTCT induced both apoptosis and necrosis of AGS and SNU-1 cells and apoptosis was the major type of cell death. Conclusion: There exists significant synergistic effect between hyperthermia and chemotherapy in inhibiting the proliferation of gastric cancer cells. Hyperthermia enhances the sensitivity of gastric cancer cells to chemotherapy. HTCT induces two patterns of cell death, apoptosis and necrosis, in gastric cancer cells. Apoptosis was the major type of cell death.

One of the major responsibilities for medical students is to save lives and to popularize emergency medical treatment knowledge for the masses of society.Based on research analysis,systemic training of cardiopulmonary resuscitation program including lectures,simulation education and social practice were performed for medical students.After practice activities,these students could proficiently apply the basic knowledge and skills of cardiopulmonary resuscitation in practical work and spread what they have learned to the public.Furthermore the program could improve students' abilities in social practice and team spirit and it is of great social significance.

It is usually a challenge to repair the abdominal wall defect to general surgeons. With developing of materials science, the use of mesh provides a novel method of primary closure of abdominal wall defects in this set-ting. Recently , biological mesh has been reported to reconstruct abdominal wall successfully. This review is to in-troduce the recent status and progress on its biological characteristics,animal experiments and clinical Study.

Objective To investigate the effects of soluble CD40 ligand combined with LY294002,a specific inhibitor of phosphatidylinositol 3-kinase,on proliferation and invasion of gastric cancer cells (AGS ) and its potential mechanisms. Methods The effect of gradient concentration of sCD40L and its combination with LY294002 on cellular proliferation was determined using MTT assay. The ability of cellular invasion and migration was evaluated by wound healing and Transwell invasion assay. The protein expression of PI3K, p-Akt and vascular endothelia growth factor (VEGF) was tested by Western blotting. Results The inhibitory effect of cellular proliferation in sCD40L, LY294002 and sCD40L + LY294002 groups was 70. 1%, 65. 7% and 41. 3%, respectively, with significant difference (P<0. 05). And wound healing was also delayed in cells treated with sCD40L plus LY294002 when compared with those treated with sCD40L or LY294002. These results indicated that the inhibitory effect of cellular proliferation, migration and invasion was enhanced when cells were treated with sCD40L plus LY294002. The protein expression of PI3K,p-Akt and VEGF increased in cells treated with sCD40L, but reduced drastically in cells treated with sCD40L plus LY294002. Conclusion Inhibiting PI3K/Akt signaling pathway may enhance the inhibitory effect of proliferation, migration and invasion by sCD40L on AGS cells, which provide a new way for biochemical therapy of gastric cancer.

Objective To study the role and mechanism of microRNA-16(miR-16)in the proliferation,invasion and apoptosis of ovarian epithelial carcinoma cells in vitro.Methods The SKOV-3 cells were transfected with miR-16 mimics or negative control RNA(NC)by lipofectamine 2000.The expression of miR-16 was detected by real-time reverse transcription(RT)-PCR in SKOV-3 cells,and western blot was used to detect the expression of vascular endothelial growth factor(VEGF),matrix metalloproteinase-2(MMP-2)and bcl-2 protein.Methyl thiazolyl tetrazolium(MTT),5-ethynyl-2'-deoxyuridine(EdU)and transwell assay were used to determine the proliferation and invasion abilities.And the rate of apoptotic cell was detected by flow cytometry method.Results(l)The expression level of miR16 in the transfection cells group was significantly higher than that in NC group(125.93 ± 15.30 versus 0.78 ± 0.16,P ＜ 0.01).(2)The rclative expression level of VEGF protein in transfection cells,NC and blank control group was 0.58 ± 0.05,1.22 ± 0.03,1.20 ± 0.03,MMP-2 protein was 0.63 ± 0.03,1.16 ±0.03,1.21 ± 0.03,and bel-2 protein 0.52 ± 0.03,1.19 ± 0.05,1.28 ± 0.06,respectively.The level of VEGF,MMP-2 and bcl-2 protein in the transfection group were lower than those in other control groups,and there were significantly differences among them(all P ＜0.01).(3)After transfected 4 days,the inhibition rate of cell proliferation in the transfection group was dramatically higher than that in NC group[(37.2 ±6.2)％ versus(3.6 ± 3.2)％,P =0.001].(4)The percentage rate of proliferative cells in the transfection,NC and blank control group was(12.3 ± 0.8)％,(23.4 ± 1.8)％,(31.1 ± 4.9)％.And it was lower in the transfection group(P ＜ 0.05).(5)Decreased cells via the transwell member in the transfection group(6 ± 3)were detected as compared with NC group(40 ± 9)and blank control group (48 ± 8,P ＜ 0.01).(6)Twenty-four hours after cultured in serum starvation and hypoxia,the rate of the viable and late apoptotic cells in the transfection group were significantly higher than those in NC group and blank control group[the rate of viable apoptotic cell was(16.9 ± 2.1)％,(10.3 ± 1.7)％ and(9.0 ±0.8)％ respectivcly,P＜0.01;the rate of late apoptotic cell was(13.4±3.3)％,(3.2 ±1.8)％ and (0.7 ±0.6)％ respectively,P ＜ 0.01].After cultured 48 hours,total apoptotic cells in the transfection group was significantly more than those in other groups(P ＜ 0.01).Conclusion miR-16 might inhibit the proliferation,invasion of ovarian epithelial carcinoma cells and enhance their sensitivity to apoptotic stimuli via downregulation of the expression of VEGF,MMP-2 and bcl-2 protein.

Objective: To study the expressions of vascular endothelial growth factor D (VEGF-D) and its receptor fms-like tyrosine kinase-4 (Flt-4) and to determine whether the presence of VEGF-D and Flt-4 was associated with the clinicopathologic characteristics and prognosis of pancreatic cancer. Methods: The expressions of VEGF-D and Flt-4 proteins were examined by immunohistochemical staining in 48 pancreatic cancer tissues, 32 adjacent pancreatic tissues, and 13 normal pancreatic tissue samples. Differences in distribution of VEGF-D or Flt4 were analyzed using the χ2 test. Kaplan-Meier survival analysis was used to estimate survival time and log-rank test was used to compare differences in survival time between the patients with or without VEGF-D or Flt-4 expression. Results: Immunohistochemical analysis revealed that the positive staining rates of VEGF-D and Flt-4 in pancreatic cancer tissues were significantly lower than that in para-cancerous tissues (P < 0.05), but significantly higher than that in normal pancreatic tissues (P < 0.05). Pancreatic cancer tissues with lymph node metastasis had higher expression of VEGF-D and Flt-4 compared with those without lymph node metastasis (P < 0.01). The median survival time and 1-, 2-, 3-year survival rate of VEGF-D-or Flt-4-positive patients were significantly decreased compared with VEGF-D- or Flt-4-negative patients (P < 0.05). Conclusion: VEGF-D and Flt-4 may contribute to lymph node metastasis and may serve as a prognostic factor for pancreatic cancer patients.