Objective To develop the detection method of common fungal pathogens by general primer PCR.Methods The primers were designed from the target genes of 5.8S rDNA and 28S rDNA of fungi,and the specificity and sensitivity were observed.Results The general primer PCR can be used to detected C.albicans,C.parapsilosis,C.krusei,C.glabrata,C.tropicalis,C.neoformans,A.fumigatus,A.flavus,A.nidulans,A.niger,C.carrionii,P.verrucosa,S.schenckii,F.pedrosoi,T.rubrum,T.mentagrophytes,M.gypseum,M.canis,E.floccosum,M.racemosus,the sensitivity was 15 pg/ml of DNA.Conclusion The general primer PCR can be used to detect common fungal pathogens.

Objective To detect the expression of hypoxia inducible factor-lα (HIF-1α) in both mRNA and protein and reactive oxygen species (ROS) levels in human epithelial ovarian cancer,and investigate the abnormal state of oxygen metabolism and their interaction in human epithelial ovarian cancer develoment.Methods The expression of HIF-1α in mRNA and protein in epithelial ovarian cancer were detected by real time reverse transcription-polymerase chain reaction and immunohistochemical Elivision plus two-step method,and the level of ROS was detected by fluorescence microplate reader.Results The relative content of HIF-1α mRNA in the epithelial ovarian cancer group was 4.466±0.754,which was higher than that of the benign control group (6.073±1.053) (t =-7.701,P =0.000).The expression of HIF-1α mRNA was raised gradually in tissues of the benign ovarian tumor,early and advanced stage of epithelial ovarian cancer (t =5.301,P =0.000).The expression of HIF-1α mRNA in the epithelial ovarian cancer had positive correlation with grade and lymph node metastasis (t =3.724,-3.851,P =0.000).The positive expression rate of HIF-1 α protein in epithelial ovarian cancer was 63.49 ％ (40/63),which was higher than that of the benign group (U =433,P =0.005),which had positive correlation with lymph node metastasis and stage (x2 =4.289,12.360; P =0.038,0.000).The expression of ROS in the epithelial ovarian cancer was 110.239±18.414,which was higher than that of the benign group (58.366±20.360) (t =11.068,P =0.000),which had positive correlation with grade,lymph node metastasis and stage (t =-2.055,2.816,-4.138; P =0.044,0.007,0.000).There was a moderate positive correlation between the expression of HIF-1α mRNA and ROS in epithelial ovarian cancer (r =0.572,P =0.000),and that was similar in low differentiation group and advanced group,there was a relatively high positive correlation in lymph node metastasis group (r =0.703,P =0.001),there was a relatively low correlation between HIF-1α mRNA and ROS (r =0.376,P =0.002).Conclusion Hypoxia is the basic characteristic of human epithelial ovarian carcers.This study suggested that HIF-1α is associated with human epithelial overaian cancer prognosis.ROS and HIF-1α might be involved in the genesis of human epithelial ovarian cancer,and the regulation of ROS to HIF-1α may play the important roles in the invasion and metastasis process of human epithelial ovarian cancer.The expression of HIF-1α is regulated by ROS at the transcriptional and protein levels,probably mainly on protein level.

AIM: To improve the prevent efficacy of peptide p277 in autoimmune diabetes. METHODS: The recombinant expression plasmid pET28-Hsp65-6×p277 was constructed by inserting 6×p277 which were amplified by PCR into the vector pET28-Hsp65. The plasmid pET28-Hsp65-6×p277 was transformed into E.coli BL21 (DE3) and the fusion protein (Hsp65-6×p277) was expressed effectively as soluble protein after inducing by lactose. The fusion protein was purified and then used to immunize 4-week old female NOD mice with three times of i.n. inoculations in the absence of adjuvants. Serum samples from the immunized mice were collected at monthly interval. The concentrations of blood glucose and antibodies were measured by automatic analyzer. RESULTS: Administration with the Hsp65-6×p277 to NOD mice could prevent the development of diabetes. CONCLUSION: The fusion protein Hsp65-6×p277 might be further developed to a vaccine against insulin-dependent diabetes mellitus.

Objective To explore the value of diffused optical tomography(DOT)with ultrasonography (US)in differential diagnosis of breast benign from malignant neoplasms.Methods One hundred ard twenty-three breast neoplasms from 113 patients were checked use the optical tomography image ultrasonography(OPTIMUS).The diagnosis results of the system were compared with pathological study.Two-dimensional ultrasound features in breast neoplasms were digitalizated,and HBT and SO_2 were detected by DOT.At last,synthesis diagnostic index(SDI)was obtained by both US and DOT.Statistic process was carried out in sensitivity,specificity,and the rate of accuracy respectively by US,DOT and OPTIMUS system.Results Among breast neoplasms of 123 mass studied pathologically,78 of them were benign and the rest were malignant.HBT in benign lesions(153.02±105.78)was significantly less than malignant lesions(232.95±78.22),SO_2 in benign lesions(1.0332±0.1641)was significantly elevated compared with malignant lesions(0.8794±0.1814),SDI in benign lesions(147.26±53.76)was significantly less than malignant lesions(243.98±57.27),significant differences were found between the two groups(P＜0.05).The sensitivity of differential diagnosis of breast neoplasms by the OPTIMUS system was 93.33%,the specificity was 83.33%,and the rate of accuracy was 86.99%.Conclusions OPTIMUS can improve the specificity and the rate of early diagnosis.

Histiocytosis, Langerhans-Cell ; pathology ; Humans ; Intestinal Diseases ; pathology

Objective To study the effects of exercise on hemodynamic parameters and cardiovascular humoral factors in the elderly patients with hypertension. Methods The modified Bruce assessment (MBA) was employed in an essential hypertension (EH) group (group 1, n =34) composed by elderly EH patients and a control group (group 2, n =31) composed of normal subjects. Heart rate (HR),systolic and diastolic blood pressure (SBP、DBP), and the quantum and duration of exercise (QE, DE) were explored, while the serum concentrations of nitric oxide (NO), endothelin 1(ET1), atrial natriuretic factor (ANF) and AngiotensinⅡ(AngⅡ) were measured at 3 min before and after treatment. Results There were no significant differences between groups 1 and 2 with regard to HR, SBP, DBP, total DE and peak QE before and after treatment, in addition, at the peak of exercise ( P 0.05). Pretreatment SBP and DBP were significantly and positively correlated with those at the peak of and after treatment ( r =0.648,0.630, P

The aim is to explore a teaching-centered management system to suit the development of clinical medicine education for international students by setting the educational aim and making training plans in combination with the present condition of the university.All the aspects of teaching such as the establishment of the teaching rules,training of the teachers,reform in teaching,teaching material as well as the teaching and student management and the service in the dorm are strengthened,so as to ensure the steady development of the foreign students education.

Teaching the foreign students clinical medicine in English in the earlier period of foreign students education has played a vital role but also has such problems as teachers,difficulties in the later teaching and so on.Through practice we find that it is necessary to transit the mode from teaching in English to bilingual education,and finally teaching in Chinese in the internship to conform with the present medical foreign students education situation.Therefore ability of teaching in English and proficiency in Chinese are needed to improved as soon as possible.

Objective To explore the effects of cognitive behavioral therapy and citalopram on regional cerebral blood flow(rCBF)in the patients with depression disorder.Methods 38 patients with depression disorder were divided into the cognitive behavioral therapy group(n=18),and citalopram group(n=20).The rCBF changes and scores of Hanmilton Depression Scale(HAMD)of all patients in two groups were evaluated with 99Tcm-ECD single photon emission computed tomography(SPECT)and HAMD before and after treatment.Results After treatment,the symptoms of all patients in two groups improved,but the cases of the citalopram group got a rapid improvement compared with those of the cognitive behavioral therapy group.Both of two methods could increase blood flow in right basal ganglion;while,the blood flow also increased in right limbic system in the cognitive behavioral therapy group,and in right temporol lobule in the citalopram group.Conclusion Both cognitive behavioral therapy and citalopram are effect on symptoms of depression disorder,but affect blood flow in different parts.

Objective To investigate the distribution and sequence conservation of genes encoding the outer membrane lipoprotein D(LPD)of nontypeable Haemophilus influenzae(NTHi)isolates and to ana-lyze the immunogenicity and the immunoprotective effects of the expressed recombinant LPD(rLPD). Meth-ods PCR analysis was used to detect the genes encoding LPD of NTHi isolates. The PCR products were se-quenced after T-A cloning. A prokaryotic expression system for genes encoding LPD was established to ex-press the rLPD. Ni-NTA affinity chromatography was used for purification. SDS-PAGE and Bio-Rad Gel Im-age Analyzer were used to detect the expression and the yield of rLPD. The antigenicity and immunoreactivity of rLPD were detected by ELISA and Western blot assay. The immunoprotective effects of rLPD against lethal dose of NTHi were evaluated in a mouse model. Results All of the tested NTHi isolates were positive for the genes encoding LPD. They shared 98. 0% -99. 4% homologies in nucleotide sequences and 98. 5% -100% homologies in amino acid sequences. The established prokaryotic expression system expressed rLPD with a high yield. High levels of antibody in rabbits were induced by the rLPD. The anti-NTHi antiserum samples from rabbits and children could recognize and react with the rLPD. The result of ELISA indicated that 93. 6%(58 / 62)and 53. 2%(32 / 62)of the serum samples from children with NTHi infection were positive for rLPD-IgM and rLPD-IgG,respectively. The rLPD at concentrations of 100 μg and 200 μg could respectively protect 60. 0% and 73. 3% of mice from lethal NTHi infection. Conclusion The genes enco-ding LPD were extensively distributed in NTHi isolates with high sequence conservation. The expressed rLPD could be used as a potential candidate antigen in the development of genetic engineering vaccine against NTHi infection considering its high immunogenicity and immunoprotective effects.