Objective To optimize the technology conditions of ultrasonic-enzyme-assisted extraction for sanguinarine from Zanthoxylum nitidum.Methods Extraction rate of sanguinarine determined by HPLC was served as an index.The applicability of the extraction solvent added with acid and enzymatic hydrolysis pretreatment to the ultrasonic-enzyme-assisted extraction of Zanthoxylum nitidum was investigated.Ultrasonic power,extraction frequency and solvent volume were optimized by orthogonal experiment.Finally,ultrasonic extraction time was optimized in dynamic process.Results The optimal process was as follows:Zanthoxylum nitidum powder was extracted 3 times by ultrasonic-wave (250 W) with 40％ ethanol (0.2％hydrochloride) as solvent (extracted for 15 rmin with 6-fold solvent at the first time,then extracted for 12 min with 3-fold solvent at the second and the third time,respectively).The extraction rate of sanguinarine was 88.6％.Conclusion The process is economic,efficient,energy-and time-saving,and provides experimental basis for industrial production of sanguinarine.

Objective To study the roles of regulated upon activation normal T cell expressed and secreted(RANTES) and monocyte cbemoattractant protein-1 (MCP-1) in human immunodeficien- cy virus-1(HIV 1) infected patients.Methods RANTES and MCP-1 in HIV-1 infected patients, including treated and untreated groups,and healthy control group were determined by enzyme-linked immunosorbent assay(ELISA).The recombinant plasmids,hMCP-pcDNA3.1,bRANTES- peDNA3.1 and hMCP/bRANTES-pcDNA3.1,were constructed and transfected into CHO cells to overexpress the corresponding recombinant proteins,whose chemoattract function was then studied. Results The level of RANTES was (164.3?21.3) pg/mL in healthy control group,(1 224.1?62.0) pg/mL in untreated group and (475.3?36.2) pg/mL in treated group.The level of MCP-1 was (90.6?28.5) pg/mL in healthy control group,(335.0?30.3) pg/mL in untreated group and (807.2?62.6) pg/mL in treated group.In HIV-1 infected patients,the levels of RANTES and MCP-1 were significantly increased.After highly active antiretroviral therapy (HAART),the level of RANTES declined,but MCP-1 increased further.Western blot assay revealed that the three recombinant proteins could be recognized by monoclonal antibodies respectively.All of them could chemoattract human peripheral blood mononuclear cells(PBMCs).And the chemoattractant potency of MCP/RANTES fusion protein was stronger.When the recombinant proteins were used with con- centrations as 50,200,400 and 800 pg/mL,respectively,the number of PBMCs chemoattracted by MCP/RANTES fusion protein was 52?10~4/mL,102?10~4/mL,132?10~4/mL and 184?10~4/mL; the number of PBMCs chemoattracted by RANTES was 27?10~4/mL,51?10~4/mL,65?10~4/mL and 96?10~4/mL;the number of PBMCs chemoattracted by MCP-1 was 18?10~4/mL,44?10~4/mL, 54?10~4/mL and 74?10~4/mL.Conclusion RANTES and MCP-1 may both be involved in the HIV infection process and host immunological reaction against HIV.

Objective To evaluate the prognostic value of microsatellite instability (MSI) in stage Ⅱ colon cancer patients.Methods 120 stage Ⅱ colon cancer patients underwent radical resection in the Department of Colorectal Surgery,Beijing Cancer Hospital from 2000 to 2007.Tissue samples were collected and DNA was extracted for MSI determination using PCR following the Pentaplex panel.Clinical parameters were also combined and analyzed statistically to explore the association between MSI status and clinical parameters.Results The incidence of high frequency of microsatellite instability (MSI-H) was 20.8％ in these 120 stage Ⅱ colon cancer patients.There was a significant correlation between the status of MSI and tumor differentiation (x2 =9.69,P =0.021);However MSI status was demonstrated to be a prognostic factor for disease free survival or overall survival (all P ＞ 0.05).Nor was MSI status associated with tumor relapse or metastasis.Age and preoperative serum CEA level were identified as independent factors for DFS by Cox regression.Conclusion In stage Ⅱ colon cancer patients,MSI status correlates with tumor differentiation,but is not a prognostic factor.

?AIM: To observe the clinical curative effect of the intravitreal injection of anti-VEGF antibody combined with the implantation of Ex-press glaucoma drainage device for neovascular glaucoma ( NG) .?METHODS:A retrospective analysis of 20 patients with NG, who got the intravitreal injection of anti -VEGF antibody combined with the implantation of Ex-press. The visual acuity, intraocular pressure ( IOP ) , iris neovascularization fade and intraoperative and postoperative complications were observed at 1wk, 1, 3 and 6mo postoperatively.?RESULTS:The average IOPs of 20 patients were 47 ± 5.6mmHg, 13.4 ±3.6mmHg, 15.3 ±4.2mmHg, 16.9 ± 5.3mmHg and 18.7 ±6.9mmHg preoperatively and postoperatively 1wk, 1mo, 3mo and 6mo with statistical difference (P<0.05).The intraoperative and postoperative complications of the implantation of Ex-press mainly included early shallow anterior chamber, drainage tube obstruction, filtering bleb scarring. There were 8 eyes with filtering bleb scarring with normal IOP.?CONCLUSION: The intravitreal injection of anti-VEGF antibody combined with implantation of Ex -press is effective for NG, which can significantly reduce the IOP.

Objective To understand the specimen sources,clinical characteristics,and antimicrobial resistance of Burkholderia cepacia (B .cepacia )isolated from infected patients in intensive care unit(ICU),so as to provide reference for guiding rational use of antimicrobial agents.Methods Clinical data of patients with B .cepacia infec-tion in an ICU between 2011 and 2014 were analyzed retrospectively,antimicrobial resistance of strains was ana-lyzed.Results A total of 267 B .cepacia strains were isolated,the major specimen sources were sputum (80.15%, n=214),blood(14.23%,n =38),and urine(3.37%,n =9).Antimicrobial susceptibility testing results revealed that B .cepacia had multiple resistance,and was naturally resistant to multiple clinically used antimicrobial agents, such as ampicillin,cefazolin,ampicillin/sulbactam,nitrofurantoin,and cefuroxime,resistant rates were all 100%;resistant rates to ceftazidime and levofloxacin were 4.12% and 3.00% respectively;resistant rate to compound sulfa-methoxazole had increased tendency(χ2 =5.885,P =0.015).Conclusion Isolation of B .cepacia in ICU increased year by year,antimicrobial resistance is serious,management and targeted monitoring of prevention and control of healthcare-associated infection should be strengthened,antimicrobial agents should be chosen according to antimi-crobial susceptibility testing results.

ABSTRACT:Objective To explore the effects of miRNA-1246 (miR-1246)on cell proliferation,invasion and migration in human cervical squamous cell carcinoma (CSCC)cell line SiHa.Methods SiHa cells were assigned into 3 groups:miR-1246 analog group,miR-1246 antagonist group and control group.Transfection efficiency was determined.The MTT assay,transwell assay and wound healing assay were performed respectively to evaluate the proliferation,invasion and migration abilities of SiHa cells.Western blot was carried out to detect the expression of thrombospondin-2 (THBS2)before and after transfection.A THBS2 3’-UTR-containing dual luciferase plasmid was synthesized and co-transfected with miR-1246 into SiHa cells to observe the luciferase enzyme activity.Results MTT assay,transwell assay and wound healing assay revealed that the abilities of proliferation,migration and invasion were significantly enhanced (P<0.01)in SiHa cells transfected with miR-1246 analog,but suppressed in SiHa cells transfected with miR-1246 antagonist.Western blot showed that SiHa cells transfected with miR-1246 analog had significantly decreased THBS2 expression (gray value = 6 .2 8 ± 1 0 .2 2 , P=0 .0 1 3 ) while those transfected with miR-1246 antagonist had significantly increased THBS2 expression (gray value = 12.90±19.81, P=0.037).After co-transfected with miR-1246 and THBS2 3’-UTR-containing plasmid,SiHa cells exhibited a decreased level of luciferase enzyme expression.Conclusion miR-1246 promoted the proliferation,invasion and migration of CSCC SiHa cell, and it might promote CSCC tumorigenesis and progression by suppressing the expression of its target gene THBS2 .

Objective:By analyzing the norm data results of the Wechsler Adult Intelligence Scale-Fourth Edition (WAIS-Ⅳ) in China,to prove the validity of the procedure and methods during the norming development.Methods:The whole process of the revision of WAIS-Ⅳ,the development of computer-assisted system and norm sampling plan,were introduced in more detail,and the distribution of actual norm data of 1757 cases was analyzed.Results:For area distribution,compared with planned sampling number,the number from North and Northeast China was statistically significant different (x2 =78.02,P ＜0.01).For age stages distribution,most of cases conformed to the requirements of sampling,except that some cases including high-level educational cases aged 16-17 years and above 65 years,and low-level educational cases aged 30-34 years were less than the planned sampling number.For gender distribution,male subjects were more,but there was no statistically significant difference between male and female subjects (x2 =228,P =0.131).For educational degree distribution,the sampling conformed to the requirements of sampling plan (x2 =2.74,P =0.603).For occupation,resident years and registered permanent residence,and the sample was basically representative.Conclusion:The process of the revision of the Chinese version of WAIS-Ⅳ is appropriate,and actual norm sampling basically conforms to planned sample distribution,providing the sufficient representativeness and reliability for national norm data of WAIS-Ⅳ.

Objective: To examine expression patterns of focal adhesion kinase (FAK) and its activated form, phosphorylated FAK (pFAK),in human osteosarcoma and to investigate the correlation of FAK expression with clinicopathological parameters and prognosis. Functional consequence of manipulating FAK protein levels was also investigated in human osteosarcoma cell lines. Methods: Immunohistochemical staining was used to detect FAK and pFAK levels in pathologically archived materials from 113 patients with primary osteosarcoma. Kaplan-Meier survival and Cox regression analyses were used to evaluate prognoses. The role of FAK in cytological behavior of MG63 and 143B human osteosarcoma cell lines was studied via the FAK protein knockdown with siRNA. Cell proliferation, migration, invasiveness, and apoptosis were assessed using cell counting kit-8, Transwell, and Annexin V/PI staining methods. Results: Both FAK and pFAK were overexpressed in osteosarcoma patients. Tumor cells exhibited cytoplasmicity and occasional membranous immunoreactivity for FAK. A total of 42 cases (37.17%) mainly showed expressed pFAK in cytoplasm of osteosarcoma cells. No overexpression staining of anti-FAK and anti-pFAK antibodies was observed in normal cancellous bone tissues or negative controls. Significant differences were observed in overall survival between FAK-/pFAK- and FAK+/pFAK- groups (P=0.016), FAK+/pFAK- and FAK+/pFAK+ groups (P=0.012), and FAK-/pFAK- and FAK+/pFAK+ groups (P<0.001). All groups showed similar metastasis-free survival. Cox proportional hazard analysis showed that FAK expression profile is an independent indicator of both overall andmetastasis-free survival. siRNA-based knockdown of FAK significantly reducedmigration and invasion of MG63 and 143B cells and affected proliferation and apoptosis in osteosarcoma cells. Conclusion: Osteosarcoma malignancies in vitro and in vivo were correlated with overexpression and phosphorylation of FAK. These findings suggest that FAK plays an important biological role in osteosarcoma carcinogenesis. This study provides a better understanding of diagnostic and prognostic relevance of FAK overexpression and phosphorylation in osteosarcoma patients. Therefore, FAK and pFAK can be used as independent predictors of overall and metastasis-free survival in osteosarcoma patients.

Objective To understand colonization of pathogens in nasal vestibular of health care workers (HCWs) in intensive care unit (ICU),and provide evidence for strengthening the prevention and control of healthcare-associated infection (HAI)in ICU.Methods On may 2015,colonization status of pathogens in nasal vestibular of uninfected HCWs in ICU were actively screened,bacterial culture,isolation and identification were performed.The surveyed results were analyzed and compared with antimicrobial resistance of pathogens from patients at the same stage.Results A total of 96 HCWs were surveyed,43 pathogenic strains were isolated from different HCWs’na-sal vestibular,isolation rate and carriage rate were both 44.79%.The main pathogenic bacteria was Staphylococcus aureus(n=15,34.88%),followed by Enterobacter aerogenes (n =9,20.93%)and Klebsiella pneumoniae (K . pneumoniae ,n=7,16.28%).There was a high detection rate of pathogens from nasal vestibular of doctors,HCWs who smoked frequently and those who never exercised (all P <0.05).There were 1 strain of imipenem-resistant K . pneumoniae among 43 pathogenic strains.Resistance rate of 7 K .pneumoniae from HCWs to ampicillin/sulbactam, cefazolin,and furantoin were all >50.00%,resistance rates to cefotaxime and imipenem were 28.57% and 14.29%respectively;resistance rates of 11 strains of K .pneumoniae from patients to furantoin was 100.00% during the same stage,but were sensitive to other commonly used antimicrobial agents.Resistance rate of 4 strains of Esche-richia coli (E.coli)to ampicillin was 75.00%,to gentamicin,tobramycin,levofloxacin,ciprofloxacin,and com-pound sulfamethoxazole were all 50.00%,6 strains of E.coli isolated from patients during the same period were found to be resistant to most commonly used antimicrobial agents.Conclusion Colonization rate of pathogens is high in nasal vestibular of HCWs in ICU,active screening and monitoring on colonization of pathogens in HCWs’ nasal vestibular is significant for preventing the occurrence and cross transmission of HAI among HCWs and pa-tients.

Several kinds of column chromatography method were used to investigate the chemical constituents of the leaves of Lophatherum gracile. The structures of the isolated compounds were identified based on their physicochemical properties and spectral data. A new flavone C-glycoside was isolated and its structure was identified as 3'-methoxyl-luteolin 6-C-beta-D-galactopyranosiduronic acid (1 --> 2) -alpha-L-arabinopyranoside (1).
Antiviral Agents ; chemistry ; isolation & purification ; Flavones ; chemistry ; Glycosides ; chemistry ; isolation & purification ; Hydrolysis ; Plant Leaves ; chemistry ; Poaceae ; chemistry