As one of important B vitamins,folate is an essential nutrient for the body and involves in various biochemical and metabolic reactions.Studies have shown that as an important methyl donor in the carbon metabolism cycle,folate has an important impact on pregnancy,pregnancy complications,and birth defects.Further studies have shown that in the folate metabolic pathway,the gene polymorphism and folic acid metabolism of a key enzyme,5,10-methylenetetetrahydrofolate reductase (MTHFR),play an important role in ovarian function.Gene polymorphism and the high homocysteine levels caused by it can lead to the damage of reproductive function and endocrine function,including follicular development,embryonic development,and hormone secretion.Gene polymorphism is also associated with the occurrence and development of ovarian disease and its response to treatment.With the deep understanding of folate metabolism,MTHFR gene polymorphism may become a new genetic marker for predicting the risk of disease and a new target for related gene therapies.

Objective A method was established for genotyping of hepatitis B virus (HBV A-D genotype),based on the PCR-restriction fragment length polymorphism (RFLP) created by Hinf Ⅰ,Ear Ⅰ,Apo Ⅰ action on an amplified segment of the S region.Methods Clinical diagnosis research.One hundred and twenty-eight HBV S sequences obtained from GeneBank were analyzed for restriction enzyme sites that would be genotype-specific.Restriction patterns following digestion with restriction enzymes Hif Ⅰ,Ear Ⅰ,Apo Ⅰ were determined to identify A-D HBV genotypes.The method was used to detect the HBV genotype of fifty severe hepatitis patients due to chronic hepatitis B in China.Then the detection results were confirmed by direct sequencing.Results The new genotyping method was established,named simple PCR-RFLP,which could identify HBV genotypes A to D.Genotypes B,C,B/C and A or D could be determined by a single step digestion with Hif Ⅰ.Eight patients of genotype A/B/C classified by single step digestion with Hif Ⅰ were conformed as genotype B variant by further digestion and direct sequencing.Extracted randomly and diluted into different concentration,three specimens were tested for genotype of HBV repeatedly and respectively.The results were all in accord with the originals,and the lowest detection limit of HBV DNA was 7 ～ 9 IU/ml.This was particularly useful in China where genotypes B and C were predominant.Twenty-three of genotype B and ten genotype C patients were classified from these fifty severe hepatitis B patients by a single step digestion with Hif Ⅰ through the simple PCR-RFLP method.The same results were also obtained by direct sequencing of PCR products (Kappa =1.00,P =0.001).The simple PCR-RFLP method was superior to direct sequencing in detecting HBV B/C polyinfection (9 cases and 0 case; x2 =18.00,P =0.001).Conclusions Both the sensitivity and repetitiveness of Simple PCR-RFLP method are satisfactory.It is superior to direct sequencing in detecting HBV B/C polyinfection,and simple,convenient.

Objective To investigate the relationship between clinical types and related risk factors in female patients with post-adolescent acne.Methods Female outpatients with post-adolescent acne aged more than 25 years were enrolled from Department of Dermatology of Renji Hospital between January and October 2016.A questionnaire survey was conducted to investigate related risk factors for post-adolescent acne in the females.Skin lesions and clinical types were evaluated by dermatologists.Statistical analysis was carried out by t test for comparison of means between two groups and by chi-square test for comparison of ratios.Results A total of 312 female patients with post-adolescent acne completed the survey,including 268 (85.9％) with mild to moderate acne and 44 (14.1％) with severe acne,241 (77.2％) with persistent acne and 71 (22.8％) with late-onset acne,or 102 (32.7％) with comedonal post-adolescent acne (CPAA) and 210 (67.3％) with papular post-adolescent acne (PPAA).Survey on related risk factors showed that 121 patients reported seasonal factors and 59 (18.9％) patients became worse in summer,and spicy,sweet and fried foods can aggravate the condition in 131 (42％),93 (29.8％) and 85 (27.2％) patients respectively.Other risk factors such as premenstrual period (62.8％,196/312),psychological factors (51.6％,161/312) and exogenous chemical exposures (43.6％,136/312) were complained of by the patients.Furthermore,premenstrual period,diet and constipation were found to be more associated with PPAA compared with CPAA (x2 =4.523,4.068,3.910,respectively,all P ＜ 0.05).Exogenous chemical exposures,such as the use of cosmetics,exposure to polluted air environment and occupational hazards,were more associated with CPAA compared with PPAA,as well as with late-onset acne compared with persistent acne (x2 =6.579,9.057,both P ＜ 0.05).In addition,premenstrual exacerbation occurred more frequently in patients with persistent acne compared with those with late-onset ache (x2 =4.512,P ＜ 0.05).Conclusions The risk factors for the occurrence of female post-adolescent acne are very complex.Premenstrual exacerbation plays a major role in the aggravation of papular and persistent post-adolescent acne,diet and constipation are more associated with PPAA,and exogenous chemical exposures are still be considered in the aggravation of comedonal and late-onset post-adolescent acne.Thus,clinical types should be considered in the diagnosis and treatment of post-adolescent acne in females.

Objective To analyze the detection rates,species distribution and drug-resistance of urinary fungal infection in elderly patients at Beijing Hospital from 2011 to 2013,in order to provide the basis for the reasonable clinical use of anti-epiphyte medicines.Methods Totally 263 patients with an average of 79.6 years old were collected from Beijing Hospital.The urine from freshly voided midstream or bladder puncture was collected under aseptic condition for fungal culture,then the strains of epiphytes were identified by using API 20C AUX.The drug sensitivity was tested with ATB fungus3.Results 263 strains of epiphytes were isolated from the 2 983 urine samples,of which 92 were C.tropicalis,85 were C.glabrata,77 were Candida albican,and 9 were other fungus candida.The rates of drug resistance to fluconazole were 14.1 ％ (13 strains),37.6 ％ (32 strains) and 15.6％ (12 strains),and to itraconazole were 16.3％(15 strains),35.3％(30 strains) and 9.1％(7 strains),respectively.All of the 263 strains were not found to have drug resistance to amphotericin.Conclusions The isolation rate of urinary fungal infections is 8.8％ in Beijing Hospital.The majority of the tested fungal are C.glabrata,C.tropicalis and Candida albican,the former has higher resistance rate to azoles,and the two latter have better sensitivity to azole,and all of them have the sensitivity to amphotericin.

Objective To evaluate clinical significance of two real-time fluorescence quantitative PCR kits for quantitative detection of HBV DNA and detection performance at different viral load levels.Methods A series of calibrators with different concentrations(1×106,5×105,1×105,5×104,1×104,5×103,1×103,5×102,1×102,1×101 kIU/L) were prepared with AB-type sera using the second generation WHO international standard (NIBSC code:97/750). HBV viral load in the sera of 78 patients,30 healthy blood donors and 10 calibrators were detected by real-time fluorescence quantitative PCR HBV DNA test kit from PIJI Bio-Technical Development Company Ltd (PG kit) and Cobas AmpliPrep/Cobas TaqMan HBV test kit. The correlation of the two methods was evaluated, and the performance of the two kits different viral load levels was evaluated. The false negative rate was analyzed. Negative control, low positive control and high positive control were included in every batch. Results Both two kits showed the correct results for the 10 specimens from the WHO international standards. The lowest detection limit of HBV DNA for Roche Cobas AmpliPrep/Cobas TaqMan HBV test and PG kit were 2.00 (kIU/L, lg) and 3.00 (kIU/L,lg) ,respectively. There was linear correlation between the results from Roche Cobas AmpliPrep/Cobas TaqMan HBV test and PG kit ( R2=0.938 7, P < 0.01 ), the upper limit of Roche kit had coincided with theoretical value. The samples with HBV DNA level above the upper limit of detection were diluted and retested to obtain the precise result. The result form Roche Cobas AmpliPrep/Cabas TaqMan HBV test [(8.35±0.20) kIU/L, lg] was higher than that from PG kit [(7.73±0.42 ) kIU/L, lg] (t=3. 776, P <0.05) . The detection of 108 serum samples showed that the level of HBV DNA detected by Roche Cobas AmpliPrep/Cobas TaqMan HBV test [(5.88±1.64) kIU/L, lg] was higher than that by PG kit [(5.25±1.55 kIU/L,lg] (t=12. 297 ,P <0.01 ). The correlation coefficients were high in samples with high HBV viral load[HBV DNA(>5.00 and≤7.00) kIU/L,Ig,R2=0. 779 7, P <0.01 ;HBV DNA( >7.00 ands≤9.00) kIU/L,lg,R2=0.603 7, P <0.01]. The correlation coefficient was low in samples with low HBV viral load[HBV DNA ( > 3.00 and≤5.00) kIU/L, lg, R2=0. 417 3, P <0.01 )]. When HBV DNA ( >3.00 and≤4.00) kIU/L,lg,the false negative rate was 33.3% (5/15). When HBV DNA ( > 1.08and≤3.00) kIU/L,lg,none of positive samples was detected with PG kit. Conclusions PG kit is not as good as Cobas AmpliPrep/Cobas TaqMan HBV test . The linear correlation between the results from the two kits is good. The correlation between the results detected with PG kit and Cobas AmpliPrep/Cobas TaqMan HBV test is higher in the high viral load groups than in the low viral load group. It is suggested that PG kit had a narrower linear range.

Objective To evaluate the effect of family interventions on the prevention of falls in elderly hypertensive patients. Methods One hundred elderly hypertensive patients were divided into the experiment group and the control group in equal number. The control group returned for regular visits after discharge while the experiment group received the family intervention including cognitive,psychological,behavioral and environmental intervention.The two groups were compared in terms of fall rate and degree of injury.Results The incidence of falls in the experiment group was significantly lower than that of the control group,the incidence of soft tissue injury after a fall in the experiment group was significantly lower than that of the control group(both P<0.05).Conclusion Family intervention is effective in prevention of falls in elderly hypertensive patients for it may reduce the incidence of falls and the degree of fall injuries.

Objective To investigate the effect of HBV on the expression of fibrosis-related factors in hepatic stellate cells(HSC)and its relation with liver fibrosis.Methods HSCs were co-cultured with HepG2 or HepG2.2.15 in vitro and HSCs cultured alone served as the control.The mRNA expression of matrix metalloproteinase(MMP)-2 and tissue inhibitor of metalloproteinase(TIMP)-1 was detected by realtime PCR.The protein expression of MMP-2 and TIMp-1 was detected by Western-blot.Results Compared with the control and the HSCs co-cultured with HepG2,the expression of MMP-2 and TIMP-1 mRNA in HSCs co-cultured with HepG2.2.15 was increased remarkably and the most significant difference was found at 72 h(F=11.91,23.13;P=0.008,0.001);the expression of MMP-2 and TIMP-1 protein in HSCs co-cuhured with HepG2.2.15 was also increased remarkably and the most significant difference was found at 72 h(F=20.70,6.54;P=0.002,0.003)too.Conclusion The expression of fibrosis-related factors in HSCs increased significantly after co-cultured with HepG2.2.15,which suggests that HBV could promote liver fibrosis.

Objective To determine the conversion equation for X(copies/ml, lg) quantity values of domestic HCV RNA quantitative fluorescence amplification assays approved by SFDA and Y( IU/ml, lg)reference values of standard substance. Methods The second generation WHO International Standard (NIBSC code:96/798) was mixed with human AB blood type serum to create 7 different dilutions which included 100 000, 50 000, 25 000, 10 000, 5 000, 2 500 and 1 000 IU/ml. Two different batches of each three domestic hepatitis C virus RNA real-time fluorescence quantitative PCR assays and 2 different batches of each assay were employed to detect the 7 different concentration samples with real-time PCR. Each test was performed 4 times repeatedly. Results The correlations between X( copies/ml,lg) values of domestic HCV RNA assays and Y(IU/ml,lg) reference values of standard substance were as follow,Assay A:Y =0. 902 0 X+0.284 9,R2 =0.953 3,P＜0. 01,n =56;Assay B: Y=0. 875 7 X +0.562 4,R2 =0.956 5,P＜0.01,n =56; Assay C: Y = 0. 843 8 X + 0. 560 5, R2 = 0. 945 8, P ＜ 0. 01, n = 56. Conclusions All the conversion equations are different among the quantity value of three assays and the reference values of standard substance, that suggests it is necessary to perform more stringent traceability analysis for the quantity values of 3 assays. Through standardizing the quantity values preliminarily, the conversion equation can enhance the comparability between the quantity values of different assays, and provide a standard of HCV RNA virus load detection for clinical diagnosis and treatment monitoring of HCV infection.

Objective To investigate the prevalence and risk factors of renal injury in chronic hepatitis C patients(CHC).Methods The clinical data of 213 CHC patients,who were hospitalized in the People's Hospital of Peking University from Jan.2002 to Oct.2007,were collected.The eGFR was caculated by MDRD equation.The prevalence and risk factors of renal injury in the CHC patients was analyzed by SPSS software.Results The patients has an average age of (53.5?14.7)years old,with male patients accounting for 59.6% and female accounting for 40.4%.We also found that 22.1% patients had hypertension,25.8% had diabetes mellitus,and 94.8% had serum positive HCV RNA.The prevalence of CKD was 26.3%,the prevalence of proteinuria was 14.6%,and the rate of hematuria was 2.8%.Serumpostive HCV RNA was the independent risk factor of proteinuria as demonstrated by multiple variation logistic regress analysis(P=0.028,OR:2.610,95%CI:1.107~6.151).Proteinuria(P=0.02,OR:3.759,95%CI:1.227~11.521),age(P=0.004,OR:1.058,95%CI:1.018~1.100)and blood uric acid(P