AimIn order to investigate the function of hsp86 gene in Plasmodium falciparum,two fragments of heat shock protein 86 gene of Plasmodium falciparum were amplified and the targeting vectors were constructed. MethodsCulturing the Plasmodium falciparum in vitro,extracting the genome DNA, amplifing the targeted gene with PCR ,contructing the target vector and identifing with restricted enzymes and sequencing. ResultsThe relative fragments were amplified successfully. The insertion and replcement vector with hsp86 gene were constructed. ConclusionConstructing successfully the replacement and insertion vector which is used to the gene knockout in Plasmodium falciparum.

Objective To evaluate the pain control, general condition and well-being of the patients with advanced cancer treated with transdermal fentanyl as well as the effects of transdermal fentanyl on heart rate variability (HRV) and endocrine function. Methods This open prospective study included 40 patients with advanced cancer. The direct conversion ratio from oral morphine to transdermal fentanyl system (TFS) was 100:1 daily. In patients who had never received any opioid before the initial dose of TFS was 25?g/h. The transdermal system was changed every 72h and the dosage was adjusted to the need of the patients according to the VAS scores and the requirement of morphine. 8 hospitalized patients were monitored by 24h Holter for changes in HRV. Blood samples were taken before transdermal fentanyl patch when pain was severe and 24h after TFS for determination of ACTH,?-endorphin, growth hormone(GH) and insulin levels.Results Pain relief during treatment with transdermal fentanyl was significant accoring to VAS scores. Well-being scores showed that patients' quality of life including appetite, sleep, mental state and daily activities was significantly improved after two weeks treatment. The ratio of LF vs HF, which is one of HRV frequency indices, was obviously reduced at 24h after transdermal fentanyl patch as compared with that before TFS (P

Objective To investigate the influence of genital tract cytomegalovirus and chlamydia infection on the incidence of tubal pregnancy. Methods Ninety five women with tubal pregnancy (study group) and 42 women with ovarian cysts (control group) were selected for this study. Serum cytomegalovirus (CMV) IgM was identified by enzyme link immunosorbent assay (ELISA). Cervical secretions, endometrium, and salpinx tissue were tested for CMV gH gene and chlamydia heat shock protein (HSP) gene by nest polymerase chain reaction (PCR) and PCR respectively. Results CMV IgM was positive in 14 (15%) women with tubal pregnancy, and 1 (2%) in the control group. CMV gH gene was detected in 18 women (19%) and chlamydia HSP gene in 25 (26%) of the tubal pregnancy group, and 2 (5%) and 2 (5%) in the control group respectively. There were significant differences between the study and the control group ( P

Humans ; Adolescent ; DNA/genetics* ; Genotype

Objective To evaluate the clinical efficacy and safety of different courses of topical tacrolimus 0.1％ ointment in facial corticosteroid-dependent dermatitis and to observe the rebound in patients after treatment with these regimens.Methods A total of 104 patients with facial corticosteroid-dependent dermatitis were randomly divided into 3 groups to be treated with topical tacrolimus 0.1％ ointment twice daily for 4,8 and 16 weeks respectively.The patients were followed up every 2 weeks within the early 4 weeks of treatment and every 4 weeks thereafter.The rebound phenomena was observed in patients on week 4 after the withdrawal of tacrolimus.Results Finally,90 patients completed this trial,including 32 patients in the 4-week group,29 patients in the 8-week group and 29 patients in the 16-week group.No significant differences were observed between the 4-,8- and 16-week groups in the total reponse rate (75.00％,82.76％,86.21％,respectively,x2 =1.35,P ＞ 0.05).The rebound rate in the 16-week group significantly differed from that in the 4- and 8-week group (20.69％ vs.46.88％ and 41.38％,both P＜ 0.05),while no statistical difference was noted between the 4- and 8-week groups.Local burning and itching were reported in 31.73％ of these patients,and all of these irritant reactions occurred within the first week of treatment.Conclusions Topical tacrolimus 0.1％ ointment is safe and effective for the treatment of facial corticosteroid-dependent dermatitis.The total response rate does not increase with the extended treatment course,and 4 weeks of treatment is enough for the marked and stable improvement of facial corticosteroid-dependent dermatitis,but the rebound rate is likely to be reduced by extended treatment course.

The B.subtilis ywtD gene,encoding a ?-polyglutamic acid(?-PGA)depolymerase,was amplified from the genome of B.subtilis NX-2 by PCR.The comparability between the cloned ywtD gene sequence to the reported sequence is high to 99.0%.Only one of the substituted nucleotide base caused the change to the amino acid sequence.The recombinant plasmid pET-15b-ywtD was then transformed into E.coli Rosetta(DE3)and the ywtD gene product could be expressed with the induction of 0.5mmol/L IPTG.The YwtD protein exhibited a remarkable activity in ?-polyglutamic acid degradation.The molecular weight of ?-PGA could be reduced from 700kDa to 20kDa after 72h through the enzymatic hydrolysis and consequently trended to be constant.

Objective To explore whether Cdc42 is an independent influence factor in regulating the migration of A549 cells by using quantitative measurement method,and to verify the effectiveness of an automatic measurement and calculation method for in vitro cell migration assay.Methods Cdc42 was overexpressed in human lung adenocarcinoma A549 cell line,and then in vitro scratch assay was applied to evaluate the migration of the cells.Different methods were used to measure the images acquired at different time points for quantitative analysis.Accuracy and repeatability of different measurement methods were analyzed.Results The results showed that overexpression of Cdc42 alone significantly advanced the migration of A549 cells (P＜0.05).The new method is efficient,accurate and reproducible as compared to manual measurement,and has significant advantages in target recognition and noise removal as compared to the existing measurement method in Image Pro Plus 6.0.Conclusions Over expression of Cdc42 significantly increased A549 cell migration ability in vitro.The new method can realize the automatic quantitative analysis of cell migration in vitro.

OBJECTIVE To compare the three methods on relatedness analysis of Acinetobacter baumannii strains in hospital infection.METHODS Twenty-seven A.baumannii strains caused hospital infection were analyzed by pulsed field gel electrophoresis(PFGE),amplified fragment length polymorphism(AFLP) and multiple gene cluster analysis.RESULTS PFGE analysis showed 19 strains isolated from clinic were with identical clone;AFLP analysis showed 19 strains isolated from clinic were with identical clone;but multiple gene cluster analysis showed 3 clones including 11 strains carried with 8 positive genes(TEM,OXA-23group,ADC,aac(3)-Ⅰ,aac(6′)-Ⅰ,ant(3″)-Ⅰ,qacE△1-sul1 and intⅠ1),6 strains with 7 positive genes(TEM,ADC,aac(3)-Ⅰ,aac(6′)-Ⅰ,ant(3″)-Ⅰ,qacE△1-sul1 and intⅠ1) and 4 strains with 6 positive genes(TEM,ADC,aac(3)-Ⅰ,ant(3″)-Ⅰ,qacE△1-sul1 and intⅠ1).CONCLUSIONS The resolving ability of multiple gene cluster analysis is higher than that of PFGE and AFLP in relatedness analysis of A.baumannii strains from hospital infection.

Objective To investigate the effect and safety of alprostadil in the treatment of diabetes complicating chronic kid‐ney disease to provide reference for clinical treatment .Methods 84 cases of diabetes complicating chronic kidney disease in this hospital from September 2013 to January 2015 were selected and divided into the observation group(44 cases) and the control group (40 cases) according to the voluntary principle .The control group used the epalrestat treatment ,while the observation group was combined with using alprostadil on the basis of control group .The effective rate ,serum creatinine ,blood urea nitrogen(BUN) ,uri‐nary albumin excretion rate ,C‐reactive protein(CRP) ,IL‐6 levels and adverse reactions were compared between the two groups .Re‐sults The effective rate of the observation group was 93 .18% ,which was significantly higher than 80 .00% in the control group , the difference was statistically significant (χ2 =4 .251 ,P=0 .005);the CRP and IL‐6 levels after treatment in the observation group were improved ,the difference was statistically significant (P<0 .05);the CRP and IL‐6 levels after treatment in the observation group[(0 .45 ± 0 .05)mg/L ,[(0 .72 ± 0 .11)μg/L] were significantly superior than[(1 .05 ± 0 .14)mg/L ,(1 .25 ± 0 .24)μg/L] in the control group ,the differences were statistically significant(P<0 .05);BUN ,urinary albumin excretion rate ,TNF‐αand endogenous creatinine clearance rate after treatment in the observation group were (6 .41 ± 1 .12)mmol/L ,(41 .12 ± 4 .46)μg/min ,(75 .54 ± 6 .64)ng/L and (92 .94 ± 8 .24)% ,which in the control group were (7 .39 ± 1 .05)mmol/L ,(91 .48 ± 7 .31)mmol/L ,(111 .42 ± 7 .69)ng/L and (81 .55 ± 9 .54)% respectively ,the improvement in the observation group was better than the control group ,the difference was statistically significant (P<0 .05);the occurrence rate of adverse reactions was 18 .18% in the observation group and 17 .50% in the control group ,showing the difference was not statistically significant (P>0 .05) .Conclusion Alprostadil in treating diabetes complicating chronic kidney disease has better effect ,conduces to improve the level of urinary albumin and inflammatory with high safety ,and is worthy of clinical promotion and application .

Objective To explore the clinical features of drug hypersensitivity syndrome(DHS). Method Retrospective analysis of 18 cases of DHS was made in detail including clinical situation, laboratory examination, treatment instance,duration and prognosis. Results The earliest symptoms of all patients were skin rash and fever, which occurred in 2 to 8 weeks after the administration of the drugs, accompanied with superficial lymphadenopathy in 13 patients, liver dysfunction in 14 patients, renal impairment in 11 patients, leukocytosis in 13 patients and eosinophilia in 10 patients. The skin lesions were multiform. The common eruption was purpura-like erythemas, accompanied with swelling and repeated desquamation. The suspected drugs were as follows: allopurinol in 11 cases, carbamazepine in 4 cases, phenytoin in 2 cases and minocycline in 1 case. Corticosteroid was the first line of treatment Conclusions DHS is a clinical syndrome to have characteristic symptom and a relatively severe disease. It is crucial to discontinue the suspected drugs promptly and to start sufficient dose of corticosteroid.