Objective To evaluate nasal carriage and antimicrobial resistance of bacteria in health care workers (HCWs)in an intensive care unit (ICU),and provide basis for making prevention and control measures of health-care-associated infection(HAI).Methods From April 2014 to March 2015,nasal swabs from HCWs in ICU were collected,carriage and antimicrobial resistance of bacteria were detected.Results A total of 450 nasal swab speci-mens were taken,137 strains were isolated,isolation rate was 30.44%.There were no significant difference in na-sal carriage rates of bacteria in HCWs with different genders,ages,types of work,length of service,and education-al level (P >0.05);nasal carriage rates in HCWs at different seasons were significantly different (P <0.05 ).82 strains (59.85%)were gram-negative bacteria,the major were Klebsiella pneumoniae (21 .16%)and Enterobacter aerogenes (18.98%);55 strains (40.15% )were gram-positive bacteria,the major were Staphylococcus aureus (18.98%)and Staphylococcus epidermidis (15.33%).38 (27.74% )strains were multidrug-resistant strains. 7.69% (2/26)of Staphylococcus aureus were methicillin-resistant strains,3.45%(1/29)of Klebsiella pneumoniae and 3.85%(1/26)of Enterobacter aerogenes were imipenem-resistant strains.Conclusion Nasal carriage rate of bac-teria and detection rate of multidrug-resistant organisms in HCWs in ICU is high.

Objective To investigate the effect of smoking and smoking cessation on the phosphorylation of IKK-β in type 2 diabetic rats. Methods Forty-two six-week-old Wistar rats were randomly divided into 4 groups: normal control(NC, n =7), diabetes control (DC, n =7), diabetes with smoking (DS, n = 14) and diabetes with smoking cessation(SC, n = 14). Rats in DS and SC groups were further assigned randomly into 8w and 12w subgroups. DS group was given passive smoking twice a day for 8 or 12 weeks, while SC group ceased passive smoking for 4 weeks after 8 or 12 weeks of smoking . Western blot method was used to detect the level of IKK-13 phosphorylation in skeletal muscle. Results Compared with the NC group,the phosphorylation of IKK-β protein in DC group was increased (0. 16±0. 05 vs 0. 30±0. 08, P < 0. 01). There was an increasing trend with the phosphorylation level of IKK-β in the DS (8w) subgroup, but there was no statistical difference between the DC group and SC(8w) subgroup (0. 40±0. 09 vs 0. 30±0. 08,0. 36±0. 10, P >0. 05). The phosphorylation level of IKK-β in DS(12w) group increased obviously, being significantly higher than that in the DC group and SC (12w) subgroup(0. 74 ± 0. 11 vs 0.30±0.08,0.35±0.07,P < 0.01). Conclusion With the prolongation of smoking duration, the phosphorylation of IKK-β in type 2 diabetic rats increased. After smoking cessation, the phosphorylation of IKK-β decreased. The phosphorylation of IKK-β may be involved in the mechanism by which smoking causes type 2 diabetes.

Objective To establish a rapid,sensitive and specific diagnostic test for detecting Neisse-ria gonorrhoea.Methods The major outer membrane proteins(P Ⅰ )in different gonococcal serogroups were obtained by isolation of outer membrane complex with CTB-ethanol precipitation,the outer membrane proteins were extracted with Z 3,14 and EDTA,and purified with DEAE-Sepharose CL-6B to obtain P Ⅰ .Hybridoma cell lines producing McAbs against P Ⅰ were established with lymphocyte hybridoma techniques.Results The molecular weight of P Ⅰ A and P Ⅰ B were determined with SDS-PAGE as35.2kDa and36.7kDa,respectively.Five hybridoma cell lines producing McAbs continuouslly and stably against P Ⅰ A and P Ⅰ B were obtained,in-cluding two hybridoma cell lines producing McAbs against P Ⅰ A and three hybridoma cell lines producing McAbs against P Ⅰ B.The titers of McAbs in the supernatants in the cultures and in abdominal ascites of BALB/c were from1:64to1:256and from1:4096to1:16384,respectively;and the specificity of the McAbs against P Ⅰ A and P Ⅰ B was so high that they easily reacted with N.gonorrhoeae but did not with other antigens such as N.meningitidis etc.Conclusion The purified P Ⅰ and the McAbs obtained in the study provide a basis to establish a rapid,sensitive and specific diagnostic test for detecting N.gonorrhoea.

The expression vector of SmIg scFv fragment was constructed in patient with B cell chronic lymphocyte leukemia (B-CLL) and expressed in E. coli to obtain scFv fragment, and the effect of the protein on the proliferation of stimulated peripheral blood mononuclear cells (PBMC) was investigated in vitro. Two pairs of primers were designed, and variable region genes of light chain and heavy chain were amplified by PCR respectively from the pGEM-T vectors previously constructed in our laboratory which containing light chain gene or Fd fragment of heavy chain gene. The PCR product was digested, purified and inserted into pHEN2 vector to construct the soluble expression vector pHEN2-scFv. After the induction by IPTG, the scFv protein was identified by SDS-PAGE electrophoresis and purified by Ni-NTA-Chromatography. MTT was used to determine the effect of purified protein on the proliferation of stimulated PBMC in vitro. Plasmid PCR and restriction enzyme digestion of pHEN2-scFv revealed the pHEN2-scFv vector was constructed successfully. Id-scFv protein was expressed in positive clone after induced by IPTG. SDS-PAGE analysis showed that the relative molecular weight of fusion protein was about 30 kD (1 kD= 0.9921 ku), which was consistent with the theoretically predicted value. Proliferation of PBMC could be induced by purified Id-scFv. It was suggested that the expression vector of SmIg scFv fragment was constructed successfully, and scFv protein was expressed and secreted from E. coli, which could induce proliferation of PBMC. This may lay an experimental foundation for further research of Id-HSP complex vaccine for B-CLL.

Objective:To analyze the implementation and effectiveness of primary health care reform in Feixi county, Anhui province. Methods: This paper uses data analysis of interviews with key insiders, policy documents and archives, as well as institutional questionnaire survey and other methods to collect information, and then it com-bines qualitative and quantitative research to arrive at research findings through cross-analysis and interpretation. Re-sults:From 2010 to 2014, the reform enhanced the capacity of primary health care in four main aspects, including capacity building of primary health care personnel, network construction of primary health care system, improvement of working conditions, and system construction of institutional management. Accordingly, the human capacity of pri-mary health care personnel was improved, their enthusiasm for work increased, their income was raised, the working conditions were improved, the cooperation mechanism for vertical and horizontal health care services was established, and the consultation rate and primary health care utilization in Feixi county increased. Conclusion:Both the“bottom-up” exploration approach and “from point to area” implementation strategy adopted by Feixi county have not only been the core content of primary health care system’s comprehensive reform, but also effectively leveraged the other aspects of the reform. They have played an important role in deepening medical reform and establishing institutional mechanism of health care system.

Objective:To analyze reverse effects induced in the implementation process of“primary health care strengthening” policies, and the innovative initiatives and countermeasures raised in primary health care system’s comprehensive reform of Feixi county. Method:Based on the principles of Critical Discourse Analysis ( CDA) , this paper designs its analytical framework for policy analysis. Results:Many reverse effects appeared in the implementa-tion process of “primary health care strengthening” policies. However, through improvements in terms of manpower upgrading, network building, working conditions and management mechanisms, Feixi county has successfully strengthened the capacity building for primary health care institutions. Conclusions:The expected and unexpected re-sults should be evaluated systematically when the policies are being implemented, and local governments need to pro-ceed according to local conditions in order to prevent unintended reverse effects from happening.

The fragmentation of health care system within a county and serious capacity constraints of health-care services in township health centers are generally the most significant problems facing underdeveloped counties in western China. Under constraints of limited public financial resources, how to increase service capability of township health centers and improve service quality, continuity and convenience of health care system has become one of the most severe challenges in implementing China’s county health care system reform. This paper takes Huangzhong coun-ty, Qinghai province as a case study. It investigates how the locals use health care alliance to vertically integrate county health care resources during reform implementation, analyzes various aspects of reform measures and the corre-sponding structural characteristics such as the integration of administrative management, integration of human re-sources, dual diagnosis referral, interoperability of information systems, sharing of resources and test results, etc. , and provides similar China’s counties with a case study to be used for reference.

Objective:After examining the changes in medical expenses and flows of hospitalized patients with-in and outside the county under the New Rural Cooperative Medical Scheme ( NRCMS ) before and after the imple-mentation of health care alliance reform, this paper analyzes the impacts of health care alliance reform on the visiting rate of NRCMS patients within the county and explores their causes and channels. Methods:This paper uses the hos-pitalization data of NRCMS patients in 2013 and 2014 collected from Huangzhong county Qinghai province and inter-views with key informants and analyzes the hospitalization expenses, visiting flows and other indicators of patients. Results:After starting health care alliance reform, the total number of hospitalized patients to seek treatment outside the county decreased by 6 . 38%, the total hospitalization expenses to seek treatment outside the county decreased by 22 . 13%, and the hospitalization expenses per inpatient day to seek treatment within and outside the county increased by 14. 41% and 20. 69% respectively. Conclusions: By improving service capabilities of township health centers and increasing synergies between county-township-village health institutions, the county health care alliance reform has suc-cessfully enhanced the service capabilities of health care systems in the whole county and reasonably helped patients with common diseases to be hospitalized outside the county and frequently hospitalized people to return back to the county.

OBJECTIVESequences analysis of Ig variable regions from the peripheral blood mononuclear cells (PBMC) of patients with chronic B lymphocytic leukemia.

METHODSTotal RNA was isolated from PBMC of patients with chronic B lymphocytic leukemia, oligo-dT-primed cDNA was synthesized from RNA. The cDNA was amplified by Taq DNA polymerase with a set of specific 5' primers corresponding to Ig FR1 and 3' primers corresponding to CH1 (C micro /C) or CL (Ckappa/Clambda), the PCR products of variable regions of Ig heavy (IgH) and light (IgL) chains were sequenced by ABI PRISM Dye terminator cycle sequencing ready reaction kit and ABI PRISM 310 Genetic Analyzer. The gene homology of variable regions of IgH and IgL chains was compared by using DNA tools 5.1 system and "the international immunogenetics database".

RESULTSFour light chains and 3 heavy chains were amplified from 4 and 3 patients respectively. Homology analysis of the sequences of 4 light chains and 3 heavy chains were performed by DNA tools system. The sequences of light chains are high homologous. And the sequences of heavy chains are quite different. The homologous analysis of the sequences of variable region by using "the international immunogenetics database" showed that the sequences were higher homologous to idiotype gene of some B lymphocytic leukemia. Four VL genes belong to human Ig Vkappa subgroup I, 2 of 3 VH genes belong to VH3 family and 1 belongs to VH5 family.

CONCLUSIONIg genes have idiotype and same disease may have same idiotype.

Base Sequence ; Cloning, Molecular ; DNA, Complementary ; chemistry ; Gene Rearrangement ; Genes, Immunoglobulin ; Humans ; Immunoglobulin Fab Fragments ; genetics ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; immunology ; Molecular Sequence Data ; Polymerase Chain Reaction

A three-dimensional (3D) graphic model of a single-chain Fv (scFv) which was derived from an anti-human placental acidic isoferritin (PAF) monoclonal antibody (Mab) was constructed by a homologous protein-predicting computer algorithm on Silicon graphic computer station.The structure, surface static electricity and hydrophobicity of scFv were investigated. Computer graphic modelling indicated that all regions of scFv including the linker, variable regions of the heavy (VH) and light (VL) chains were suitable. The VH region and the VL region were involved in composing the "hydrophobic pocket". The linker was drifted away VH and VL regions. The complementarity determining regions (CDRs) of VH and VL regions surrounded the "hydrophobic pocket". This study provides a theory basis for improving antibody affinity, investigating antibody structure and analyzing the functions of VH and VL regions in antibody activity.