Two pairs of primers specific to small subunit ribosomal DNA of Plasmodium falciparum were designed and the expected SSUrDNA fragment was amplified for detecting P.falciparum infection with double-ternperature-nested polymerase chain reaction using DNA prepared by boiling method. The results showed that the nested PCR could amplify a constant size of desired SSUrDNA fragment of P. falciparum which was further confirmed by digestion of restrlction endonuclease and could detect parasitemia level of 0. 8 ? 10-6. It has great potentials for identifying Plasmodium species in ring form of erythrocytic stage and detecting mixed Plasmodium infections. Therefore, it is suggested that this method is sensitive, accurate, simple and rapid in detecting Plasmodium falciparum in blood samples for malaria diagnosis.

AIM:To construct hybrid nucleic acid vaccines which contain Plasmodium falciparum merozoite surface protein 1 (MSP1 ) block 1 7gene and gene fragment coding for several T cell epitopes. METHODS:MSP1 block 1 7gene of FUP strain was connected with a polyvalent gene fragment which contains several T cell epitopes from MSA1 ,MSA2 ,RESA,IL- 1 and Tetanus toxin,the connected gene fragment (hybrid gene,HG) was cloned into eukaryotic expression plasmid p CDNA3.1 (- ) ,VR1 0 1 2 for intracellular expression,VR1 0 1 2 /TPA for secretion,the recombinants were identified by PCR and restriction enzyme digestion. RE- SUL TS &CONCLUSION:The hybrid nucieic acid vaccines:VR1 0 1 2 /HG,VR1 0 1 2 /TPA/ HG and p CDNA3.1 /HG were successfully constructed.

Objective] To explore the humoral and cellular immune responses in mice to eukaryotic expression recombinant plasmid encoding histidine rich protein 2 (HRP\|Ⅱ) of Plasmodium falciparum. [Methods] The start and stop codes were introduced into HRP\|Ⅱ gene fragment, the reading frame and the position of start and stop codes in HRP\|Ⅱ were identified by sequencing. HRP\|Ⅱ fragment containing the start and stop codes was cloned into pcDNA3 1(\|) to form pcDNA3 1(\|)/HRP\|Ⅱ. The BALB/c mice were immunized i.m. with the plasmids for 3 times in 3 weeks intervals. Two weeks after the last immunization, the sera and splenocytes were collected to investigate anti\|HRP\|Ⅱ antibodies by ELISA and the splenocytes proliferation response to HRP\|Ⅱ. [Results] Sequence data show that the reading frame and the position of start and stop codes are correct. Restriction enzyme digestion indicated that the HRP\|Ⅱ gene fragment containing start and stop codes was successfully cloned into pcDNA3 1(\|). Mice raised significant anti\|HRP\|Ⅱ antibodies after pcDNA3 1(\|)/HRP\|Ⅱ immunization, and the splenocytes proliferated prominently when stimulated with HRP\|Ⅱ protein. [Conclusion] Eukaryotic expression recombinant plasmid \{encoding\} HRP\|Ⅱ gene can induce significantly humoral and cellular immune response in mice. HRP\|Ⅱ gene may be a good candidate for P.falciparum blood\|stage multiple DNA vaccine.

Aim To investigate the anti-arrhythmic effect and mechanism of ?-opioid receptor during myocardial ischemia and reperfusion in rats,and to initially determine the regulation of U50488H(U50,a selective ?-opioid receptor agonist) to angiotensinⅡ(AngⅡ),endothelin(ET) and nitric oxide(NO) in rats.Methods Rats were randomly divided into 7 groups,i.e.,control group,ischemia/reperfusion group(I/R),U50488H+I/R group,PTX group(PTX,a Gi/o proteininhibitor),Glib group(glibenclamide,a K_(ATP) channel blocker),Che group(chelerythrine,a selective PKC inhibitor),and Gen group(Genistein,a Tyrosine kinase inhibitor) respectively.The arrhythmia occurrence and score in different groups were observed and counted.The contents of AngⅡ,ET and NO in plasma of rats were also examined.Results ① Compared with I/R group,the arrhythmia score of U50+I/R group was significantly decreased.The effect of pared with I/R group,the arrhythmia score of U50+I/R group was significantly decreased.The effect of glibenclamide and chelerythrine respectively,the anti-arrhythmic effects induced by U50488H in the rats during myocardial ischemia and reperfusion were significantly attenuated or even completely blocked.③ The anti-arrhythmic effects of U50488H were not significantly affected by pretreatment with genistein.④ In comparison with normal rats,the contents of AngⅡand ET in plasma of I/R group were significantly increased,but the content of NO was decreased.With the administration of U50488H,the contents of AngⅡand ET in plasma of rats in U50488H+I/R group were significantly decreased.Meantime the content of NO was increased.Conclusions ① U50488H-induced anti-arrhythmic effects in the rats with myocardial ischemia and reperfusion are mediated by ?-opioid receptor.The signaling pathway may be related with(Gi/o,) PKC,and K_(ATP) channel.② The activation of ?-opioid receptor may elicit anti-arrhythmic effect through the down-regulations of AngⅡ or ET and up-regulation of NO in plasma of rats.

Objective:To explore the etiology of male urethral stricture,analyze the therapeutic strategies of urethral stricture,and summarize the complicated cases.Methods:The data of 183 patients with urethral stricture were retrospectively analyzed,including etiology,obstruction site,stricture length,therapeutic strategy,and related complications.Results:The mean age was 49.7 years,the average course was 64.7 months,and the constituent ratio of51 to 65 years old patients was 38.8％ (71/183).The traumatic injury of patients accounted for 52.4％ (96/183),in which the pelvic fracture accounted for 35.5％ (65/183) and the straddle injury accounted for 16.9％ (31/183).There were 54 cases of iatrogenic injury (29.5％).The posterior urethral stricture accounted for 45.9％ (84/183),followed by the anterior urethral stricture (44.8％,82/183) and the stenosis (6.6％,12/183).A total of 99 patients (54.1％) received the end to end anastomosis,and 40 (21.9％) were treated with intracavitary surgery,such as endoscopic holmium laser,cold knife incision,endoscopic electroknife scar removal,balloon dilation,and urethral dilation.In the patients over 65-years old,the urethral stricture rate was 14.8％ and the complication rate (70.4％) for transurethral resection of the prostate (TURP) was significantly higher than that of all samples (P＜ 0.01).Conclusion:Both the etiology of male urethral stricture and the treatment strategy have changed and the incidence of traumatic and iatrogenic urethral stricture has increased in recent 3 years.The main treatment of urethral stricture has been transformed from endoscopic surgery into urethroplasty.

Objective:To translate the health literate healthcare organization 10 item questionnaire(HLHO-10) into Chinese and examine its reliability and validity.Methods:The Chinese version of HLHO-10 questionnaire(HLHO-10-C) was developed by following the Brislin translation model of translation, back translation, cultural adaptation and questionnaire epistemological survey.Five experts and 1 071 medical staff from 24 healthcare organizations in Zhejiang province were selected to conduct the validity and reliability test of the HLHO-10-C.Results:The content validity indices at the item level and total questionnaire level of HLHO-10-C were from 0.8 to 1.0 and 0.96 respectively, and the results of the exploratory factor analysis showed good structural validity.Conclusions:HLHO-10-C proves adequate reliability and validity to serve as a tool for healthcare organizations in evaluating and becoming HLHO. It can also help the implementation of the Healthy China Initiative(2019—2030), which is a performance assessment mechanism for health education and promotion of healthcare providers and health care organizations.