Objective To study the molecular identification method of Candida species and lay the foundation of molecular diagnosis for deep candidiasis. Methods PCR was used to amplify the internal transcribed spacer region of ribosomal DNA from 11 type strains of 9 pathogenic Candida spp. and 39 clinical Candida isolates, and the products were digested with three types of restriction endonucleases (MspⅠ,DdeⅠand HaeⅢ). Results Five patterns of products with different molecular weights were obtained after amplification from 9 Candida spp., and the products were digested with MspⅠ, DdeⅠand HaeⅢ, producing 8, 5 and 4 unique band patterns, respectively. Conclusion PCR-RFLP is reliable, specific and useful for the identification of common pathogenic Candida species.

Objective To study the molecular identification method of Sporothrix schenckii based on the nucleotide sequences in internal transcribed spacer region 2 of ribosomal DNA (rDNA) genes. Methods Species-specific primers were used to amplify the internal transcribed spacer region 2 of rDNA of 22 clinical isolates of Sporothrix schenckii and 12 strains of dematiaceous fungi. Totally 11 strains of Sporothrix schenckii were sequenced and analyzed, in which 1 strain came from the US and the others were isolated from different areas in China. A pair of species-specific oligonucleotide primers (SSP) were designed based on the ITS2 sequence. With the species-specific primers, rDNA of Sporothrix schenckii and dematiaceous fungi were amplified by PCR. Results Sequencing and analysis showed that internal transcribed spacer region 2 of rDNA gene was conservative in Sporothrix schenckii. A 300 bp fragment was obtained from 22 strains of Sporothrix schenckii, but not from the other species. Conclusions This method is specific, sensitive and reliable for the identification of Sporothrix schenckii and could be used for clinical molecular diagnosis.

Objective To study the risk factors of severe hand-foot-mouth disease (HFMD) among children.Methods The clinical data of 1 570 children with HFMD at Linyi People's Hospital in Shandong Province in 2011 were collected,retrospectively.The data were analyzed using univariate and multivariate Logistic regression.Results The mean age of severe HFMD (including severe and critical HFMD) was (25.0± 14.0) months old,predominantely aged between 1 and 5 years old,while mild HFMD was (27.1±15.8) months (t'=-2.717,P=0.007).There were 61.0％ and 65.9％ boys in two groups,respectively (x2 =3.894,P=0.048).Fever,convulsion,tremor,nausea and vomiting were more frequently seen in severe HFMD.The neutrophil count and the level of creatine kinase in severe HFMD were both significantly higher than that in mild HFMD.Univariate analysis revealed that age (odds ratio [OR]=1.799,95％CI:0.984-1.997),girl sex (OR=1.234,95％CI:1.001-1.522),high fever (OR=2.110,95％CI:1.816-2.452),convulsion (OR=1.878,95％CI:1.578-2.236),nausea and vomiting (OR=1.760,95％CI:1.456-2.128),neutrophil count (OR=1.031,95％CI:1.025-1.037) and creatine kinase (OR=1.002,95％CI:1.001-1.003) were risk factors for severe HFMD.Multivariate Logistic regression showed that high fever (OR =1.751,95％ CI:1.487-2.062),convulsion (OR=1.451,95％CI:1.204-1.749),nausea and vomiting (OR=1.269,95％CI:1.027-1.568),neutrophil count (OR=1.028,95％CI:1.021-1.035) were independent risk factors.Conclusions Body temperature,neurological manifestations and trend of neutrophil counts should be carefully monitored in children with HFMD.Prevention of the development of severe HFMD mainly relies on the identification of risk factors and adoption of precautions in time.

Objective To investigate the clinical efficacy of levosimendan injection in the treatment of patients with acute decompensated heart failure.Methods70 patients with acute decompensated heart failure patients were selected from January 2016 to January 2017 in huzhou first people's hospital.35 cases of patients in control group were treated with levosimendan injection, and the other 35 cases of patients in experimental group was treated with dobutamine.BNP, 24 hourly urine volume, LVEF and SV were detected and the systemic clinical conditions were assessed both before and after the treatment.ResultsAfter treatment, all indexes of two groups were better than before(P<0.05).The extent of improvement of systemic symptoms and relevant indicators such as BNP, 24 hourly urine, LVEF and SV in experimental group was larger than that in control group (P<0.05).Adverse reaction rate of experimental group was 16.7%, and adverse reaction rate of control group was 43.8%, the differences were statistically significant (P<0.05).ConclusionLevosimendan injection can relieve symptoms of patients with acute decompensated heart failure,and it has less adverse reactions.

OBJECTIVE: To investigate the features of adverse drug reaction(ADR) in our hospital and evaluate the quality of ADR report in order to promote rational clinical drug use and find out the existing problems in the report. METHODS: Microsoft Excel was designed to analyze statistically the sex and age of the patients,category of the used drugs and the clinical manifestations of ADR in 311 cases of ADR report collected between 2002 and 2005.RESULTS: The occurrence of ADR induced by antibacterials ranked the first.ADR affecting skin and its appendages counted for 50% of the total.The numbers of ADR reports were found to be less than expected.CONCLUSIONS: Professional staff training should be strengthened,the use of antibacterials should be rationalized to lessen and avoid the occurrence of ADR.

Objective To evaluate high resolution manometry in the diagnosis of hiatal hernia.Methods Clinical data were reviewed on 20 patients suffering from gastroesophageal reflux who had laparoscopic Toupet fundoplication for preoperative tentative diagnosis of hiatal hernia.Preoperative diagnosis of hiatal hernia was made collectively by endoscopy,X-ray examination,24 hour esophageal pH monitoring and high resolution manometry before surgery.Results Preoperative diagnosis of hiatal hernia was made in 3 patients by X-ray examination,in 9 patients by high resolution manometry.11 patients were finally diagnosed with hiatat hernia intraoperatively.X ray was consistent with intraoperative diagnosis in 27％ cases.Intraoperative and endoscopic diagnoses were 55％.High resolution manometry and intraoperative diagnoses were consistent in 82％.Lower esophageal sphincter length was (1.92 ± 0.38) cm in hiatal hernia group and (2.10 ± 0.92) cm in non-hiatal hernia group (t =0.60,P ＞ 0.05),lower esophageal sphincter pressure (respiratory min) was (0.64 ±0.55) kPa in hiatal hernia group and (1.31 ± 1.07) kPa in nonhiatal hernia group(t =1.80,P ＞ 0.05),and lower esophageal sphincter pressure (respiratory mean) was (1.43 ±0.92) kPa in hiatal hernia group and (2.57 ± 1.33) kPa in non-hiatal hernia group(t =2.26,P ＜0.05).The reflux parameters,including the percent total time pH ＜ 4,and DeMeester score,were significantly greater in hiatal hernia group than in non-hiatal hernia group (all P ＜ 0.05).Conclusions Hiatal hernia patients are with poor esophageal antireflux competency and severe reflux.High resolution manometry is more valuable in the diagnosis of hiatal hernia than endoscopy or X-ray examination.

Objective To assess the distribution of vaginal microflora in healthy women and investigate their cognition of lower reproductive tract infection-related knowledge as well as personal hygiene habits and character of behavior in seeking medical treatment. Methods Total of 1660 healthy women who had physical check-ups at the Medical Center of Renji Hospital were selected and received gynecologic examination, as well as routine examination, pH examination and bacterial culture of the vaginal discharge. In addition, 860 of them were randomly selected for questionnaire survey, in which reproductive tract infection -related knowledge and personal hygiene habits as well as behavior in seeking medical treatment were involved. Results Among the 1660 eensused women, vaginal average pH was 4. 16±0.21. The positive rate of Candida in vaginal discharge routine examination was 3.86% (64/1660), which was lower than that in aerobes culture 7.71% (128/1660). Candida alhicans was the most populous species 78.9% (101/128) ,followed by 7.8% (10/128) and 7.0% (9/128) for the Candida glabrata and Candida krusei respectively. The most populous species of vaginal microflora were hemolytic streptococcus A (63.80%, 1059/1660), Staphylococcus epidermidis (14.28%, 237/1660), and Enterococcus faecalis (D) group ( 11.44%, 190/1660). The questionnaire survey showed that 88.4% (760/860) of 860 women took active treatment when feeling unwell, 92.1% (792/860) of them had good hygiene practices, and only 21.2% (182/860) had the habit of vaginal douching. In addition, 50.0% (430/860) of them had the desire to obtain reproductive health knowledge through out-patient consultation. Conclusions Vulvovaginal Candida disease ranks the first in all types of vaginitis, among which, Candida albicans is the most populous species followed by the Candida glabrata and Candida krusei. Hemolytic streptococcus A and Staphylococcus epidermidis are the most common species of vaginal mieroflora in healthy women. The censused women have high awareness of reproductive health care and pay mueh attention to common gynecologic diseases such as lower reproductive tract infection. Nevertheless, we should strengthen the public education of reproductive health-related knowledge.

Objective:To investigate the role and action mechanism of aquaporin 3 (AQP3) in skin photoaging.Methods:Normal human skin fibroblasts (NHDF) were divided into several groups: NHDF group receiving normal culture without transfection, AQP3 cDNA group transfected with AQP3 cDNA, AQP3 siRNA group transfected with AQP3 siRNA, heterogeneous nuclear ribonucleoprotein Q (hnRNPQ) cDNA group transfected with hnRNPQ cDNA, hnRNPQ siRNA group transfected with hnRNPQ siRNA, AQP3-hnRNPQ cDNA group transfected with AQP3 and hnRNPQ cDNAs, AQP3-hnRNPQ siRNA group transfected with AQP3 and hRNPQ siRNAs, cDNA empty vector group transfected with a cDNA empty vector, and siRNA empty vector group transfected with a siRNA empty vector. Transfected or untransfected NHDFs were irradiated with ultraviolet A (UVA) at a dose of 10 J·cm -2·d -1 for 3 consecutive days to establish a model of cellular senescence, and NHDF receiving no UVA irradiation served as a control. A cell counting method was used to evaluate the cellular proliferative activity, a senescence-related β-galactosidase staining kit to evaluate the senescence level of NHDFs in each experimental group, and luciferase reporter gene technology to assess the transcriptional regulation activity of p53. Western blot analysis was performed to determine the expression of AQP3, hnRNPQ and senescence-related proteins p53 and p21 in NHDFs. Two-independent-sample t test was used for comparisons between two groups, and one-way analysis of variance for comparisons among multiple groups. Results:After 3-day consecutive irradiation with UVA, the expression of p53 and p21 in NHDFs and the percentage of β-galactosidase-positive cells significantly increased compared with the unirradiated control group (all P < 0.05) , but the expression of AQP3 and cellular proliferative activity on days 5, 6 and 7 significantly decreased in the UVA group compared with the unirradiated control group (all P < 0.05) . After 3-day consecutive irradiation with UVA, aggravated senescence-related phenotypes of UVA-induced NHDFs were observed in the AQP3 siRNA group compared with the siRNA empty vector group, and there were significant differences in the expression of p53, p21 and hnRNPQ, percentage of β-galactosidase-positive cells, p53 transcriptional regulation activity and cellular proliferative activity between the 2 groups (all P < 0.05) . Further silencing of the hnRNPQ gene could reverse the above effects. Compared with the siRNA empty vector group, the senescence-related phenotypes of UVA-induced NHDFs were attenuated in the hnRNPQ siRNA group, and significant differences were observed between the 2 groups in terms of the expression of p53, p21 and hnRNPQ, percentage of β-galactosidase-positive cells, p53 transcriptional regulation activity and cellular proliferative activity (all P < 0.05) . After 3-day consecutive irradiation with UVA, the senescence-related phenotypes of UVA-induced NHDFs were significantly attenuated in the AQP3 cDNA group compared with the cDNA empty vector group (all P < 0.05) , manifesting as significantly decreased expression of p53 (0.25 ± 0.06 vs. 0.56 ± 0.08) , p21 (0.23 ± 0.06 vs. 0.70 ± 0.07) and hnRNPQ (0.82 ± 0.09 vs. 0.92 ± 0.03) , percentage of β-galactosidase-positive cells (31.23% ± 6.54% vs. 81.53% ± 7.62%) and p53 transcriptional regulation activity (2.52 ± 0.36 vs. 7.16 ± 0.25) , but increased cellular proliferative activity ([2.93 ± 0.33]× 10 6/ml vs.[2.15 ± 0.23]× 10 6/ml) , and further overexpression of hnRNPQ could reverse the above effects. After 3-day consecutive irradiation with UVA, the expression of p53, p21, percentage of β-galactosidase-positive cells, p53 transcriptional regulation activity and cellular proliferative activity in the hnRNPQ cDNA group were 1.41 ± 0.09, 1.42 ± 0.06, 91.06% ± 4.24%, 12.35 ± 0.88 and (1.23 ± 0.41) × 10 6/ml respectively, and the senescence-related phenotypes of UVA-induced NHDFs were significantly aggravated in the hnRNPQ cDNA group compared with the cDNA empty vector group (all P < 0.05) . Conclusion:AQP3 may alleviate the UVA-induced senescence of NHDFs by regulating hnRNPQ and downregulating p53 expression.

Objective The purpose of this study is to explore the clinical value of CT angiography( CTA) in the diagnosis of multiple intracranial aneurysms.Methods The data of CTA and DSA from 74 patients with multiple intracra-nial aneurysms from July 2011 to March 2015 were reviewed retrospectively.Results One hundred seventy-seven aneu-rysms were detected by DSA, and 175 aneurysms were detected by CTA.Among the aneurysms identified by CTA, 4 aneu-rysms were false positive and the correct detection rate of CTA was 96.6%.One hundred sixty-five aneurysms identified by CTA were confirmed by DSA and the correct diagnostic rate of CTA was 96.5%.CTA failed to detect 6 aneurysms and mis-diagnosed 10 aneurysms.Conclusions The correct detection rate and diagnostic rate of CT angiography ( CTA) in multiple intracranial aneurysms is relatively high.But previous surgery, spasm of the vessels, the size and number of aneurysms, radiologists'experience can influence the accuracy of ( CTA) in the diagnosis of multiple intracranial aneurysms, indicating that we should combine CTA with DSA to avoid the misdiagnosis and missed diagnosis.

Objective To evaluate the detection of alpha 5 (Ⅳ) collagen chain of skin basement membrane in diagnosis of Alport syndrome among suspected patients. Methods Data of suspected patients with the detection of alpha 5 (Ⅳ) collagen chain of skin basement membrane were retrospectively collected and analyzed from January 2007 to March 2008. Results A total of 254 suspected patients ranged from 1 to 71 years old with an average age of (25.85±17.03) years old were enrolled (male/female ratio, 0.76). There was no significant difference in average age between male and female. Abnormal alpha 5 (Ⅳ) collagen chain expression of skin basement membrane was found by indirect immunofluorescense in 19 patients among whom 12 cases were negative and 7 cases were diseontinous deposit. These 19 patients were diagnosed as Alport syndrome and the diagnostic rate was 7.5%. Conclusions The diagnostic rate of Alport syndrome by detection of alpha 5 (Ⅳ) collagen chain in skin basement membrane is significant and helpful for early and differential diagnosis of Alport syndrome.