Laboratory diagnostics is one of the most fast developing medical sciences.But the teaching quality falls behind due to the traditional education model.Based on the concept of ‘three-stage fusion' of the subject and ‘five abilities' of the training objects,‘three-highlight & stereo' teaching style was established and put through the course,which means to emphasize function,practice,effect in optimizing teaching content and methods,and construct stereo platform including information system,online course,laboratory equipments,research activities and teacher training program.The results showed a significant improvement of the students' knowledge mastering and utilizing capability.And the teaching situation was well re-created.Furthermore,the teaching team was much more powerful than ever.It suggests that ‘three-highlight and stereo' teaching style is a successful explore and practice of teaching reform of laboratory diagnostics on adapting for internationalization of education tendency.

Objective: To clone human carboxypeptidase A1 and its active center gene as well as construct recombinant vector for expression before analysis their activities. Methods: CPA1 and CPAlactive center gene were amplified by RT-PCR from pancreas tissue, and then sequencing was carried out. The correct target genes were cloned into prokaryotic vector pGEX-4T-1 and transformed into E. coli BL21 before sequence analysis. After induced by IPTG, gene products were analyzed by SDS-PAGE. Target expressed proteins were denaturated, renaturated, purified and evaluated through MTT and agar colony form test. Results: Human carboxypeptidase Al and its active center gene were cloned successfully. New expected protein band of Mr 66,000 and 46,000 appeared on SDS-PAGE after inducement. Both of the expressed proteins have catalytic activity in vitro, but the activity of the latter is inferior when applied to tumor cells. Conclusions: Human carboxypeptidase A1 and its active center gene were cloned successfully. Their prokaryotic expression products were obtained too. The expressed proteins have catalytic activity in. vitro. A new prosperous beginning of further improvement for CPA1 therapy system has been established based on CPA1 and its active center gene in terms of ADEPT against prostate cancer to clinical application.

Objective To observe the clinical effects of dermis combined autologous transplants in patients with abnormal scar thickness skin graft.Methods 13 cases of scar deformity were selected during August 2011 to December 2014 in our hospital.In stage 1 procedure,the scars that affected the function and induced deformities were resected and then the artificial dermis covering the wound was placed;in stage 2,two weeks after the artificial dermis vascularization,autologous skin grafts were removed to repair the wound edge.Meanwhile,therapy of inhibiting scar after conventional treatment was conducted with funcitional exercise.Results 13 cases were scheduled to repair scar wounds,underwent surgery with more than 6 months follow-up observation.The dysfunctions caused by scar contracture and deformity were significantly improved;appearance,elasticity and appearance of the skin grafts were similar to autologous skin graft with the desired clinical effect.Conclusions Artificial dermis combined autologous skin graft treatment of patients with scar deformity can achieve similar clinical effects to traditional treatment methods.

Objective:To know about the prevalence rates of depression and anxiety and its related factors a-mong human immunodeficiency virus (HIV)-infected men who have sex with men (MSM)in China.Methods:Under the help of MSM group,totally 600 HIV-infected MSM were recruited by snowballing sample among HIV-positive MSM,and 541 effective electronic questionnaire were received.Participants completed self-made question-naire of HIV-positive MSM healthy condition which included social demographic and health behavior et al.The Center for Epidemiologic Studies Depression Scale(CES-D)and Self-Rating Anxiety Scale (SAS)were used to survey the condition of depression (CES-D score ≥16)and anxiety (SAS score ≥50)respectively.Results:The percentage of having depression and anxiety symptoms was 44.7% (242 /541)and 25.1% (136 /541)respective-ly.Multiple unconditioned logistic regression analysis showed that the risk of having depressive symptoms included considered suicide in the past year (OR =3.31,95%CI:1.73 -5.34),nondisclosure their HIV infection to male primary sexual partners (OR =0.47,95%CI:0.30 -0.73),discrimination experience (OR =1.19,95%CI:1.11 -1.29).Compared with monthly income of 1500 yuan or less,participants who earned >5000 yuan monthly were less likely to have depressive symptoms (OR =0.34,95%CI:0.17 -0.71).Factors associated with anxiety symp-toms included considered suicide in the past year (OR =2.43,95%CI:1.40 -4.21),accepted antiviral therapy (OR=1.74,95%CI:1.06 ~2.84),discrimination experience (OR =1.12,95%CI:1.05 ~1.20).Participants earned >5000 yuan monthly (OR =0.44,95%CI:0.20 ~0.94)could decrease the risk of having anxiety symptom compared with whose monthly income of 1500 yuan or less.Conclusion:The prevalence rate of depression and anxiety symp-tom among HIV-infected MSM is high and need more attention.It may be helpful to call for the strategy of promo-ting mental health on decreasing social discrimination,communicating with sexual partners,and improving their eco-nomic status.

Objective To construct fusion gene and prokaryofie expression plasmid encoding the Rv1884c and Rv0867c genes in Mycobacterium tuberculosis (M.tb).The fusion protein wsg expressed efficienfly in E.coli cells.Methods The Rv1884c and Rv0867c genes were amplified by polymerase chain reactions (PCR) with specific primers from genomic DNA of M.tb H37Rv strain,and cloned into pGEX-4Tland pUC19 vectors.Rv1884c and Rv0867c were subcloned into the expression vector pGEX-4T-1 and pPRO-EXHT followed by DNA sequencing.The plasmids were transformed into E.coli DH5α and induced to produce GST-fused Rv1884c and His-fused Rv0867c fusion protein.The protein molecular weight and expression format was analyzed by SDS-PAGE.Results The recombinant expressive vectors pGEX-4T-1Rv1884c and pPRO-EXHT-Rv0867c were constructed.The DH5α strains of E.coli with recombinant plasmid showed high level of Rv1884c and Rv0867c gene expressions after IPTG induction.The SDS-PAGE showed that the plasmids expressed Rv1884c and Rv0867c fusion proteins with molecule weight of 45 000 and 80 000.The recombinant protein accounted for 18.3%and 23.7%of total bacteria protein.The expressed proteins could be purified via GSTrao FF and Ni2+-NTA system kits in denatured condition.Conclusions Mycobacterium tuberculosis Rv1884e and Rv0867c genes have been cloned and expressed Successfully in E.coli DH5α.The results lay a basis for further study of fast culfivation in Mycobacterium tuberculosis and investigation of their activities and functions.

Objective To investigate the value of Fluorine-18 fluorodeoxyglucose positron emission tomography (~(18)F-FDG-PET)in staging and detecting residual disease during follow-up of non-Hodgkin's lymphoma in children. Methods The results of~(18)F-FDG-PET of 88 podiatric NHL before and after chemotherapy was analyzed respectively.The results were compared with CT scans simultaneously.Results 67 cases were performed for initial staging.Of the 1072 anatomy regions were analyzed by PET and CT with concordalice of 88.62%regions(positive in 11.10%and negative in 77.52%1.Disconcordance were found in 11.38%regions(PET positive but CT negative in 6.8 1%,PET negative but CT positive in 4.57%).PET was significantly better than CT scan in the staging of podiatric NHL.35 scans were performed in the follow-up.The true residual disease positive rate of PET and CT were 87.50%and 57.14% respectively.The false positive rate of PET and CT were 12.50%and 50.00%respectively.Conclusion ~ (18)SF-FDG-PET imaging is of great value for the staging of the pediatric patients with NHL It's a useful technique to identify the residual disease,avoid unnecessary treatment and biopsy and Can predict eady relapse.

Objective To explore the risk factors of osteoporosis and the relation with pulmonary dysfunction in elderly patients with chronic obstructive lung disease (COPD). Methods One hundred and eighty patients (82 females and 98 males) with acute exacerbation of chronic obstructive lung disease (AECOPD) from March 2006 to June 2008 were selected in the study. The bone mineral density (BMD) of lumbar vertebrae and hip joint were determined by dual energy X-ray absorptiometry(DEXA). All the patients were divided into two COPD groups with and without osteoporsis. The smoking history, incidence of vertebral fractures, glucocorticosteroid using condition and so on were recorded. The pulmonary function, 6-minute walk distance(6MWD), body mass index (BMI) and serum albumin concentration were evaluated. Results The mean age of all patients was (72±7)years, and the average smoking amount was (59±27)pack years. The ratio of forced expiratory volume in 1 second to forced vital capacity (FEV1/FVC) was(36.46±9.8)%, and 30% of the patients had inhaled or oral glucocorticoids for more than 3 months. The BMD measurement results showed that BMD of 95% patients(171 cases) was lower than the normal level, and 119 cases (66%) had osteoporosis, including 61 males and 58 females (62%vs. 70%, x2 = 1.435, P=0.33), and 52 cases had (29%) osteopenia. Linear correlation analysis showed that BMI, 6MWD, RV% and FVC% had positive correlation with osteoporosis (r=0.362, 0.635, 0.688, 0.973;all P<0.05).Conclusions The prevalence of osteoporosis is high in elderly patients with moderate or severe COPD, and enough attention and active intervention shoule be paid.

Objective To detect the isoniazid (INH) and rifampin (RIF) resistance of Mycohaeterium tuberculosis isolates in the single tube with multiplex-polymerase chain reaction-single strand conformation polymorphism(muhi-PCR-SSCP) system. Methods According to the sequences of inhA, katG and rpoB genes of the Mycohacterium tuberculosis, three pairs of oligonucleotide primes were designed to examine the INH and RIF resistance with the multi-PCR-SSCP. The validity of the newly developed method was evaluated with 116 clinical isolates of Mycohacterium tuberculosis( 70 isolates that were INH-resistant and 66 isolates that were RIF-resistant). Results The validity of the method was assessed with multiplex PCR-SSCP with the bacteria culture with susceptibility test as golden standard. The three genes, katG, inhA and rpoB, in the 116 clinical isolates and H37Rv strain were amplified successfully in single PCR reactions,except 4 isolates with katG deletion mutants. Compared with strain H37Rv, forty-six isolates had katG gene mutations, thirteen had inhA mutations and fifty-eight had rpoB mutations. Thirty-eight isolates had simultaneous katG and rpoB mutations and 4 isolates had both inhA and rpoB mutations. Four isolates had inhA and katG mutations and 2 isolates had mutations in all three genes simultaneously. The sensitivity of the newly developed multiplex-PCR-SSCP assay was 80% and 82% for INH and RIF, respectively. The specificity of the assay was 100% and 92% for INH and RIF, respectively. Conclusion Muhiplex-PCRSSCP provides a rapid, specific and cost-effective method of detecting multidrug-resistant TB. It laid a solid foundation for the further study of drug resistant gene.

Objective: To estimate the direct medical cost of severe hand, foot and mouth disease (HFMD) in patients aged less than five years. Methods: A stratified sampling method was used to collect data on severe HFMD cases reported in the National HFMD surveillance database between Jan 1, 2012, and Dec 31, 2013. The sampling was referenced with the national aetiologic distribution of Enterovirus A71 (EV-A71), Coxsackievirus A16 (CV-A16) and other Enteroviruses (OEV) for severe HFMD cases and the included cases were distributed among seven geographic regions (Northeast, North China, Northwest, Central China, Southwest, East China and South China). A nationwide telephone interview using a structured questionnaire was conducted to obtain the direct medical cost and any complications that occurred in patients during the outbreak of laboratory-confirmed HFMD. After excluding the cases who could not recall their medical expenses or complications, a total of 685 cases were included in the analysis. Kruskal-Wallis H test was used to analyze the differences among patients who reported different complications. Multiple linear regression with bootstrap analysis of 500 replicates was used to explore the factors that influenced the direct medical costs. Results: Of 685 patients analyzed, 456 (66.6%) were male and 229 (33.4%) were female. The direct medical costs P₅₀ (P₂₅, P₇₅) were 14 250 (10 301, 20 600) Yuan. In total, 127 (18.5%) patients were diagnosed with severe HFMD patients with respiratory disease, 38 (5.5%) patients were diagnosed with aseptic meningitis, and 378 (55.2%) with encephalitis/brainstem encephalitis/acute flaccid paralysis. Furthermore, 53 (18.5%) patients were diagnosed with myocarditis, 39 (5.7%) with pulmonary hemorrhage/pulmonary edema and 50 (7.3%) with cardiopulmonary failure. The median (interquartile range) direct medical costs were 12 360 (7 313, 16 480) Yuan for severe HFMD patients with respiratory disease, 13 803 (9 064, 19 930) Yuan for aseptic meningitis, 14 438 (11 000, 20 015) Yuan for encephalitis/brainstem encephalitis/acute flaccid paralysis, 14 800 (8 500, 21 218) Yuan for myocarditis, 20 600 (12 500, 31 130) Yuan for pulmonary hemorrhage/pulmonary edema, and 20 043 (12 772, 28 840) Yuan for cardiopulmonary failure (H=17.70, P<0.001). The results of multiple linear regression with bootstrap analysis revealed that the direct medical cost for severe HFMD patients from Central China was 7 881 (95% CI: 3 814-11 949) Yuan higher than that of North China; severe HFMD patients diagnosed with OEV had direct medical costs of 1 987 (95%CI: 206-3 769) Yuan less those associated with EV-A71; severe HFMD patients whose duration of illness was ≥21 d had 20 480 (95% CI: 10 985- 29 974) Yuan higher direct medical costs those whose illness lasted ≤5 d; the direct medical costs for severe HFMD patients with pulmonary hemorrhage/pulmonary edema and cardiopulmonary failure were 7 874 (95%CI: 3 723-12 026) and 9 855 (95% CI: 328- 19 382) Yuan higher, respectively, than that associated with respiratory disease. Conclusion The direct medical costs associated with severe HFMD were found to be substantial. The total cost was affected by the duration of the illness and the severity of any complications.

OBJECTIVETo understand the coverage of HIV related intervention and antiviral treatment among HIV-positive men who have sex with men (MSM) and to examine factors of initiating antiretroviral treatment.

METHODSA total of 501 HIV-positive MSM were recruited by "snowballing" sampling in Chengdu (160), Chongqing (168) and Guangzhou (173) from January to June, 2013. Participants who were older than18 years, had sex with men, HIV-infected, were living in these 3 cities when survey was conducted were eligible for subjects of this survey. The data were collected by computer assisted survey, including social demography, the coverage of HIV related follow-up intervention and ART, related knowledge level, sexual behaviors in the last 6 months, as well as notification to sexual partners and family. We analyzed the influence factors for initiating ART by Multiple Unconditioned Logistic Regression.

RESULTSAmong 501 HIV-positive MSM, the ratio of CD4 counting and HIV viral load testing were 90.8% (455) and 19.4% (97), respectively. The percentage of the latest CD4 counting not more than 350 per µl was 33.0% (150/455). The coverage of initiated ART among the participants who met the qualification was 56.0% (84/150). The multivariable Logistic regression analysis showed that the possibility of not on ART among participants who were migrants increased to 5.21(2.33-11.66) times of the local residents and the possibility among participants who were diagnosed STD in the last year increased to 2.70(1.12-6.55) times of those who were not infected STD, the possibility of not on ART among participants who had sex with male occasional or commercial partner in the last 6 months increased to 2.16(1.03-4.50) times of those who hadn't, and the possibility among participants who had anal sex with condom use in the past 6 months increased to 2.68(1.10-6.50) times of those who couldn't insist using condom.

CONCLUSIONThere were low coverage of series of HIV/AIDS related intervention services among HIV-positive MSM in Chengdu, Chongqi and Guangzhou. Migrants, had sex with male occasional or commercial partner, had anal sex with condom use in the past 6 months, not infected STD in the last year may be the independent risk factors for HIV-positive MSM to not initiating ART.

Anti-Retroviral Agents ; therapeutic use ; CD4 Lymphocyte Count ; China ; Cities ; Condoms ; Demography ; Emigrants and Immigrants ; Follow-Up Studies ; HIV Infections ; drug therapy ; epidemiology ; Health Knowledge, Attitudes, Practice ; Homosexuality, Male ; Humans ; Logistic Models ; Male ; Risk Factors ; Risk-Taking ; Safe Sex ; Sexual Behavior ; Sexual Partners ; Surveys and Questionnaires