Objective To study impact of traditional Chinese medicine liquid combined with compression method on wound healing of anal fistula after towing line operation. Methods 120 patients with anal fistula were divided into observation group and control group by random number table including 60 cases respectively. Observation group were given liquid washing fistula combined with sand bags oppression in wound on the basis of conventional nursing; Control group were given the routine nursing care. Curative effect, pain, wound healing time, recurrence rate and degree of satisfaction of two groups were compared. Results Clinical curative rate of observation group was 98.33% (59/60), which was obviously higher than 85.00% (51/60) of control group, and there was statistically significant difference (χ2=6.981 8, P<0.01). Postoperative pain score at the 1st, 2nd, 3rd day of observation group was (2.98± 2.01), (2.97±1.94), (3.06±1.92) points, which was lower than (5.48±1.76), (5.52±1.91), (5.42±2.01) points (t=13.336 9, 13.514 8, 12.261 3, P < 0.01). Wound healing time of patients in observation group was (23.96±3.28) days, which was obviously less than (32.34±4.51) days of control group (t=6.285 1, P<0.01);Recurrence rate in observation group was 1.67%(1/60), which was obviously lower than 11.67%(7/60) of the control group, and the difference was statistically significant (χ2=4.821 4, P < 0.05). Conclusions Applying traditional Chinese medicine liquid combined with compression method in wound healing of anal fistula after towing line operation can promote wound healing, improve curative effect, prevent recurrence, and improve patients′satisfaction.

Estrogen participates in many life activities through combination with estrogen receptor alpha (ERalpha) or estrogen receptor beta (ERbeta) in the body. In order to establish an in vitro estrogen-like compound screening model, the coding region of human ERalpha and ERbeta was separately constructed into pET32-ERalpha and pET43-ERbeta prokaryotic expression vector and water-soluble recombinant ERalpha and ERbeta proteins were expressed in Escherichia coli strain BL21. Western blotting revealed that both recombinant proteins have estrogen receptor binding sites. The proteins were purified using S-Tag affinity Purification Kit and digested with enterokinase to get the ERalpha and ERbeta proteins. About 0.90 mg of ERalpha and 0.65 mg of ERbeta were obtained at the concentration of 0.181 and 0.131 mg x mL(-1), respectively.

Objective To explore the risk factors of chronic cough in children.Methods A hospital-based case-control study was conducted.A total of 60 children with chronic cough and 120 non-chronic cough children were interviewed with standard questionnaires.Non-conditional multivariable logistic model was performed to analyze the risk factors.Results Among 192 children that were performed questionnaire survey,180 cases were obtained the complete data with a questionnaire response rate of 93.75％.No significant difference in age,gender,and permanent residence was found between chronic cough and control groups,respectively (P ＞0.05).As shown in multivariable logistic model,parents with sensitive history (OR =1.924),mother or father smoking (mother:OR =1.989 ; father:OR =2.156),poor ventilation (OR =27.906),and interior decoration less than 3 months stay (OR =4.652) increased the risk of chronic cough in children.Conclusions Many factors,even the domestic environmental factors,are associated with chronic cough in children.It's time to strengthen the intervention of risk factors for reducing the occurrence of chronic cough in children.

Objective To investigate the inhibitory effects of L. monocytogenes expressing melanomainhibiting activity(MIA)gene on malignant melanoma.Methods The plasmid pERL3-hly-MIA was constructed and used to transform live L.monocytogenes by electroporation.Atier culture.the transformed L.monocytogenes,namely LM-hly-MIA,was harvested for the detection of MIA protein by Westem Blot.A total of 24 mice were divided into three groups,namely control group,LM group and LM-hly-MIA group,to be immunized with physiological saline,L.monocytogenes and LM-hly-MIA,respectively.One day after the immunization.mice were subcutaneously inoculated with 0.1 mL of B16 cells at a concentration of 1×107/mL.One week later.reimmunization was performed.The size and weight of tumors were observed and measured in these mice.Results Enzyme digestion and Western blot confirmed the Successful construction of plasmid pERL3-hly-MIA.The average weight of tumors in mice was 4.33±0.91 gram.3.36 ±0.41 gram and 1.89±0.52 gram,respectively in the control group,LM group and LM-hly-MIA group,respectively(P＜0.05).with the inhibition rates of tumor growth being 22.4%and 61.5%in LM group and LM-hly-MIA group.respectively.Conclusion The attenuated strain of L.monocytogenes expressing MIA gene could evidently inhibit melanoma growth.

Objective To investigate possible role of 14 -3 -3 in the pathogenesis of asthma inflammation, the expression of 14 -3 -3 protein was observed in lung tissues of asthmatic rats.Methods Expressions of 14 -3 -3 protein was determined by immunohistochemisty method in lung tissues,and the relationship between 14 -3 -3 and asthma inflammation was analyzed.Results The location of positive expression of 14 -3 -3 protein was mainly at cytoplasm,while little at plasmalemma.The positive expression cell mostly was bronchial epithelial cell,others were lymphocytes,alveolar macrophages and vascular endothelial cells.On the other hand,the bronchial smooth muscle and vascular smooth muscle were negative expressed.Moreover,the expression level of 14 -3 -3 protein in asthma group [(0.353 ±0.023)absorbance]was significantly higher than that in the control group[(0.211 ±0.028 )absorbance] (t =10.969,P <0.01).Conclusion The results showed that the 14 -3 -3 protein was overexpressed in asthmatic lung tissue,it may play an important role in asthma inflammation through bronchial epithelial cells,lymphocytes, alveolar macrophages and vascular endothelial cell.

AIM: To establish an HPLC method for determing content of puerarin isoflavones in effective parts of Naomaitong Granules.METHODS: Gradient elution with methanol-water-glacial acetic acid was used as the mobile phase.The flow rate was 1.0 mL/min,and the detection wavelength was set at 250 nm.RESULTS: The average recoveries of puerarin,daizein-8-capiosy(1→6) glucoside,3-methoxyl-puerarin,daidzin,daidzein were 100.89%(RSD = 2.1%),100.23%(RSD = 1.06%),101.04%(RSD = 1.92%),99.82%(RSD = 2.02%),102.06%(RSD = 1.34%) respectively.CONCLUSION: The method is simple,accurate and can be used for quality control of the effective parts in Naomaitong Granules.

Objective To investigate the inhibitory effect of live-attenuated Listeria monocytogenes (LM)-based vaccines expressing the gene encoding a melanoma differentiation antigen,MART-1,on malignant melanoma and their mechanism.Methods The constructed plasmid pERL3-MART-1 was used to transform live-attenuated LM by electroporation to construct recombinant LM.i.e.△inlB LM-MART-1 and △actA/△inlB LM-MART-1.The half lethal dose (LD50) of attenuated listeria strains was determined by concentration gradient dilution method.C57BL/6 mice were randomly divided into three groups,namely PBS group,△inlB LM-MART-1 group and △actA/△inlB LM-MART-1 group.Mice were inoculated by intraperitoneal injection of O.1 LD50 of each rLM strain or PBS only.One week later,the mice were injected subcutaneously with 1×105 B16F10 cells(a mouse melanoma cell strain)in 200μl of PBS.Reimmunization was performed on day 14 and 21.Subsequently,the growth of tumor and survival of tumor bearing mice were observed.All mice were killed on day 28,and tumor tissue as well as splenocytes were obtained from these mice for the detection of MART-1 gene expression by real-time quantitative PCR and the percentage of CD4+CD25+T cell by flow cytometry.Results The recombinant △inlB LM-MART-1 and △actA/△inlB LM-MART-1 were constructed successfully.The LD50 of △inlB LM and △actA/△inlB LM was lower than LM-EGDe by 100 and 10 000 times respectively.Compared with PBS,the tumor growth was inhibited with △inlB LM-MART-1 by 46.95%(F=6.3,P<0.05),and by 83.96% with △actA/△inlB LM-MART-1(F=37.8,P<0.01).The relative expression level of MART-1 in △inlB LM-MART-1 group and △actA/△inlB LM-MART-1 group was 8.988±0.207 and 11.315±0.445 times that in PBS group (both P<0.05).The percentage of CD4+CD25+T cells in splenocytes was (2.52±0.20)%,(1.14±0.13)% and (0.44±0.15)% in PBS group,△ialB LM-MART-1 group and △actA/△inlB LM-MART-1 group,respectively;the differences were statistically significant between the three groups (all P

BACKGROUND:In recent years, based on high-throughput molecular imaging, integration of genomics, proteomics and computer aided design and the application of correlative “technical chains” have achieved great achievements in the research of breast cancer, lung cancer, gastric cancer, colon cancer, ovarian cancer and melanin tumor. However, there are few researches on oral squamous cel carcinoma. OBJECTIVE:To detect the gene expression profile of the oral squamous cel carcinoma tissue and normal paracarcinoma tissue using DNA chip-based gene expression profile. METHODS:Two samples of oral squamous cel carcinoma tissue and normal paracarcinoma tissue of patients who received treatment at Stomatological Hospital of Guangdong Province of China in 2013 were included in this study. The gene expression profiles of oral squamous cel carcinoma and normal paracarcinoma tissue were determined by the Roche NimbleGen gene expression microarrays. RESULTS AND CONCLUSION: According to screening criteria of differential genes, 7 872 out of 32 448 detected genes were differentialy expressed genes of oral squamous cel carcinoma, which accounts for 24% of the total number of the screening genes. 3 800 genes were up-regulated, and 4 072 were down-regulated. The results confirm that through detection with the help of gene expression profile clip, 7 872 differentialy expressed genes were obtained through DNA chip-based gene expression profiles according to the screening criteria. Thus it can be concluded that the occurrence and development of the tumors are not a result of single or several genes. Previous experiments based on a single or several genes have great limitations. These findings also suggest that the occurrence of tumor is a result of mutual regulatory effects of many genes forming a network, moreover, the interactions of the network is quite complicated.

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Listeria monocytogenes (LM), a Gram-positive facultative intracellular bacterium, can be used as an effective exogenous antigen expression vector in tumor-target therapy. But for successful clinical application, it is necessary to construct attenuated LM stain that is safe yet retains the potency of LM based on the full virulent pathogen. In this study, attenuated LM and recombinants of LM expressing melanoma inhibitory activity (MIA) were constructed successfully. The median lethal dose (LD(50)) and invasion efficiency of attenuated LM strains were detected. The recombinants were utilized for immunotherapy of animal model of B16F10 melanoma. The level of MIA mRNA expression in tumor tissue was detected by using real-time polymerase chain reaction (PCR) with specific sequence, meanwhile the anti-tumor immune response was assayed by flow cytometric analysis and enzyme-linked immunosorbent spot (ELISPOT) assay. The results showed the toxicity and invasiveness of attenuated LM were decreased as compared with LM, and attenuated LM expressing MIA, especially the double-genes attenuated LM recombinant, could significantly induce anti-tumor immune response and inhibit tumor growth. This study implicates attenuated LM may be a safer and more effective vector for immunotherapy of melanoma.