Centering on the reform practice in Sanming featuring a synergistic reform in public health services,medical insurance and medicine production-circulation (Tripartite-sector reform below),this paper focused on the top-level design of the management system in such a reform and its synergistic operation mechanism.The authors probed into the main reasons for its grounding-breaking success and challenges,to pave way for their discussions on the reproducibility and sustainability of the reform.

Objective To compare the performance of fourth generation HIV antigen/antibody combined detection reagents for HIV early infection samples,international HIV seroconversion panel samples and routine clinical screening samples.Methods Thirty seven early HIV infected samples from the followup gays in Shen Yang between 2009 and 2011,66 seroconversion panel samples from BBI company (U.S.A),NABI company(U.S.A) and NIBSC company(U.K) and 703 routine HIV screening samples in the first hospital of China medical university in October 2010 were collected.All kinds of samples were tested by three diagnostic reagents based on chemiluminescence assay (CLIA),electrochemiluminescence assay (ECLIA) and enzyme-linked immunosorbent assay (ELISA) respectively.The detection sensitivity and specificity of these assays were analyzed.Results For 59 early infected and seroconversion samples,the sensitivities of both ECLIA and CLIA reagent were 96.61％ (95％ CI 91.5％-100.0％),higher than that of the ELISA kit (95％ CI 75.0％-92.9％) (x2 =5.341,P ＜ 0.05),which is 83.93％ ; Comparison among the three reagents for different subtypes of the antibody seroconversion samples showed that ECLIA had the highest sensitivity while CLIA was the lowest ; Detection sensitivity of the three reagents for the P24 antigen is CLIA ＞ ECLIA ＞ ELISA; With detection of 703 clinical routine screening samples,the specificities of three reagents were 100％ (CLIA),99.86％ (ECLIA) and 99.71％ (ELISA) respectively.Conclusions For the sensitivity of the fourth HIV diagnostic reagents CLIA and ECLIA are better than ELISA.The former two reagents are more suitable for identifying earlier HIV infection in clinic.

Objective To investigate the possible association of HLA-DQ alleles with pelvic inflammatory disease (PID) caused by Chlamydia trachomatis in Han nationality in Northern China. Methods Thirty five Han Chinese patients with PID caused by Chlamydia trachomatis and 98 unrelated healthy individuals from the same area were genotyped for HLA-DQ gene alleles by the PCR-sequence specific oligonucleotide (SSO) method. Results Compared with the healthy controls, the frequencies of HLA-DQA1*0501 and HLA-DQB1*0301 allele were significantly increased and also associated with the antibody response to Chlamydia trachomatis heat-shock protein 60 (CHSP60). Conclusions These results may provide clues to reveal the susceptibility gene of chlamydial pelvic inflammatory disease in China and the immunogenetic mechanisms of the disease.

Objective To evaluate the detectability of HIV antigen-antibody in the window period of acute infection by three HIV antigen-antibody assays.Methods Twenty-two samples of HIV seroconversion serum panels and thirty-seven HIV acute infected plasm samples from our laboratory collected from cohort study of men who have sex with men between 2009 and 2011,were assayed by ECLIA,CLIA and ELISA methods.All assays were evaluated for the ability to detect HIV in the window period,and the sensitivity of each assay for acute samples was analyzed.Chi square test was used for statistical analysis.Results The ability of detecting HIV in the window period of each assay was different.For HIV seroconversion serum panels,the results of ECLIA and CLIA assays were consistent,and the window period was shortened at least 1 to 5 days compared with ELISA assay.For HIV acute samples,all were HIV positive by ECLIA or CLIA assay,but for ELISA assay,94.6％ was positive.For samples before seroconversion,ECLIA and CLIA assay had the same sensitivity (93.5％),which is superior to ELISA assay (71.0％) (x2 =5.14,P ＜0.05).Conclusion The ability of detecting HIV in the window period was different for each assay.The results of ECLIA and CLIA assay are consistent,superior to ELISA assay.

ObjectiveTo screen mimetic HIV-1 neutralizing epitopes from plasma with high level neutralizing antibody,and to provide useful information for further study of the interaction between antigen and antibody.MethodsIn order to gain neutralizing antibody recognized mimotopes, we detected neutralizing antibodieslevelsof 11HIV-1infectedslowprogressorsbyPBMC-basedneutralization assays.High-titer HIV-neutralizing antibodies from plasma of SPs was used as the ligand for biopanning by phage-displayed random peptide library.Positive phage clones was evaluated by ELISA,sequenced,and analyzed for homology to HIV-1 env by local BLAST to deduce the neutralizing peptide.ResultsTwenty-two clones were obtained consistent with requirement through three rounds biopanning.After comparison analysis,twelve clones include C8 were obtained as mimotopes of neutralizing antibody,C40 located in gp41Ⅱ cluster with the highest titer by inhibition ratio may be as neutralizing epitope.Conclusion By the use of IgG antibodies from SPs to screen the phage random polypeptide library,one can acquire multiple phage mimetic peptides of HIV related antigen epitope.(Chin J Lab Med,2012,35:838-842 )

Objective To investigate the level of B7-H1 and its counter-receptor PD-1 expression in mDC and different subsets of T lymphocytes in HIV infected individuals in China and to analyze the correlation between the level of B7-H1/PD-1 and disease progression, and to demonstrate that PD-1/PD-L1-dependent inhibition is operating in HIV infected patients.Methods Percentage of B7-H1 and PD-1 expression in mDC, CD+4 T cells and CD+8 T cells from thirty-six untreated HIV infected patients and 20 health controls were selected and detected by flow-cytometry, its correlations with CD+4 T cell absolute counts and plasma viral loads were analyzed.Results The percentage of B7-H1 expression in mDC, CD+4 T cells and CD+8 T cells (mean 15.21, mean 20.63, mean 13.5)were higher than that of health controls (all P<0.05).The percentage of PD-1 expression in CD+4 T cells and CD+8 T cells (mean 17.91, mean 19.21)were higher than that of health controls (P<0.05, P<0.05). The level of B7-H1 and PD-1 expression were inversely correlated with CD+4 T-cell counts(mDC+B7-H1+:r=-0.647, P<0.01;CD+4B7-H1+:r=-0.489, P=0.002;CD+8B7-H1+:r=-0.372, P=0.026;CD+4PD-1+:r=-0.374, P=0.025;CD+8PD-1+:r=-0.455, P=0.005) and positively correlated with HIV viral load(mDC+B7-H1+:r=0.662, P<0.01;CD+4B7-H1+: r=0.426, P=0.01;CD+8B7-H1+:r=0.531, P=0.001;CD+4PD-1+:r=0.362, P=0.03;CD+8PD-1+:r=0.380, P=0.022).Conclusion The level of B7-H1 and PD-1 expression was associated with HIV disease progression, which provides a useful marker to define disease progression of HIV infection.

Objective To evaluate the role of hippocampal neuronal nitric oxide synthase(nNOS)-postsynaptic dense protein 95(PSD95)coupling in short-term memory retrieval disorder induced by sevoflu-rane in mice.Methods Sixteen clean-grade healthy Kunming mice of both sexes,aged 2-3 months,weighing 30-35 g,were divided into 2 groups(n＝8 each)according to the random number table method:sevoflurane group(S group)and nNOS-PSD95 uncoupling agent ZL006 group(Z group).After successful establishment of dark avoidance memory,3.3％sevoflurane and 40％O2 were inhaled for 2 h in both groups,and normal saline 1.5 ml was intraperitoneally injected in group S and ZL006 1 mg/kg in group Z at 30 min before anesthesia.The step-through latency and error times were recorded before anesthesia and at 12 h after the end of anesthesia.The mice were then sacrificed,and hippocampal tissues were taken for de-termination of the expression of nNOS and PSD95(by Western blot)and co-expression of nNOS and PSD95(by immunoprecipitation).Results Compared with that before anesthesia,the step-through latency was significantly shortened,and the error times were increased at 12 h after anesthesia in group S(P＜0.05),and no significant change was found in the above indicators in group Z(P>0.05).Compared with group S,the step-through latency was significantly prolonged,error times were decreased,the co-expression of nNOS and PSD95 was down-regulated(P＜0.05),and no significant change was found in the expression of nNOS and PSD95 in group Z(P>0.05).Conclusion The mechanism by which sevoflurane induces short-term memory retrieval disorder may be related to promoting the coupling of nNOS to PSD95 in the hippocam-pus of mice.

Objective To explore the polymorphism of nef gene and conservation level of functionally important domains of nef as well as their influences on HIV-1 disease progression of HIV-1 B'infected individuals in northern China.Methods 30 long term nonprogressors(LTNPs)and 42 typical progressors (TPs)were selected.Provirus DNA was extracted from whole blood sample.The full nef gene was amplified by nested-PCR.PCR product was sequenced directly after purification.Phylogenetic analysis and amino acid sequence mutation was applied on nef sequences to explore the differences between LTNPs and TPs.Results At position 15,the S15R/K/N substitution was detected.The frequency of TPs(64.29%)wsa higher than LTNPs(33.33%,P<0.01,0R=3.60);R21K/E/H/I.Q,TPs and LTNPs mutation frequency was 59.52%and 93.33%(P<0.005,OR=0.11);At position 39,K39R/E/N was only detected in TPs(23.81%,P<0.005).Conclusion No significant deletion or defect associated with disease progression wsa detected in nef gene of HIV-1 B'.But it suggested that K/E/H/I/Q mutation at 21 st amino acid of nef associated the disease nonprogression.R/K/N at 15 th amino acid of nef and R/E/N mutation at 39th amino acid of nef associated the disease progression in.HIV-1 B'.All domaias of nef amino acids sequences were comparatively conservative.

Objective:To test the expression of Minichromosome maintenance complex component 7(MCM7) protein in hepato-cellular carcinoma(HCC) of different species including human, rat and tree shrew (tupaia) by cross-species oncogenomics approach, and to investigate the relationship between the expression of MCM7 and the development of hepatocellular carcinoma and its clinical significance. Methods:Western blot and Immunohistochemistry were applied to detect the expression levels of MCM7 protein in HCC tissues,corresponding HCC-adjacent liver tissues and normal liver tissues collected from different species including human, rat and tree shrew, respectively. The clinicopathologic factors were also analyzed with the results of Immunohistochemistry. Results:Western blot analysis showed that the expression of MCM7 protein in HCC tissues of human and rat were higher than that in corresponding HCC-ad-jacent liver tissues and normal liver tissues, respectively and significantly (P<0.05). However, the expression of MCM7 protein in HCC tissues of tree shrew were also higher than that in corresponding HCC-adjacent liver tissues and normal liver tissues, but no significant difference was found among three types of tissues (P>0.05).There was also no significant difference between HCC-adjacent liver tis-sues and normal liver tissues in three species (P>0.05). Immunohistochemical analysis showed that MCM7 protein was mainly ex-pressed in nucleus of HCC cells, and the positive rate of MCM7 protein in HCC tissues of human, rat and tree shrew were significantly higher than that in corresponding HCC-adjacent liver tissues and normal liver tissues, respectively (P<0.05). However, no significant difference was found between HCC-adjacent liver tissues and normal liver tissues (P>0.05). Moreover, the protein level of MCM7 was intimately related to patient's HCC stage, extrahepatic metastases and postoperative recurrence (P<0.05). Conclusion:MCM7 protein might play a pivotal role in hepatocarcinogenesis. In addition, it was probably related to patient's HCC stage, extrahepatic metastases and postoperative recurrence. It seems very likely that MCM7 may be applied as a new molecular target in HCC prevention and treat-ment.

AIM To study the effects of Rehmanniae Radix Preparata (A) and Poria (B) on decoction amounts of loganin,morroniside and 5-hydroxymethylfurfural in Corni Fructus (C).METHODS These three medicinal materials were combined one another and divided into seven groups (A,B,C,A + B,A + C,B + C and A + B + C).Then the contents of three constituents were determined by HPLC.RESULTS Compared with the single decoction of Corni Fructus,the decoction amounts of loganin and 5-hydroxymethylfurfural were decreased,and that of morroniside was increased at the time of mixed decoction of Rehmanniae Radix Preparata and Corni Fructus,or Rehmanniae Radix Preparata,Poria and Corni Fructus.This situation was just the contrary at the time of mixed decoction of Poria and Corni Fructus.CONCLUSION The mixed decoction of Corni Fructus,Rehmanniae Radix Preparata and Poria increases the decoction amount of morroniside,which may make mixed decoction liquid show better efficacy.