Objective To establish the quantitative detection of capsule associated protein 10 (CAP10)and virulence-associated DEAD-box RNA helicase 1(VAD1)genes in Cryptococcus neoformans (CN) and compare the diagnostic values of single gene test and combined gene test in CN meningitis.MethodsTwenty-three CN meningitis patients with fungal culture or ink staining or CN antigen detection positive in cerebrospinal fluid (CSF) were recruited and patients with craniocerebral trauma were recruited as controls.Standard plasmids were constructed using standard CN strain.Real time fluorescent quantitative polymerase chain reaction (RT-FQ-PCR) was established to detect the mRNA expressions of CAP10 and VAD1 genes in the CSF of patients with CN meningitis,which were compared with the results of CSF ink staining,fungal culture and antigen detection.The diagnostic values of single gene test and combined gene test were compared by chi square test.Results Among 23 CN meningitis patients,22 (95.6％) were CAP10 mRNA positive detected by RT-FQ-PCR,which was significantly higher than both ink staining (16/23,69.6％,x2 =4.167,P＜0.05) and fungal culture (15/23,65.2％,x2=5.143,P＜0.05),respectively; but not significant different from antigen detection (21/23,91.3％,x2=0.500,P＞0.05).There were also no statistical significant differences between combined detection of CAP 10 + VAD1 and CAP 10 or VAD1 single gene test (P＞0.05).ConclusionRT-FQ-PCR detection is successfully established using virulence genes as target,which is superior to the conventional methods.

Objective To determine the level of awareness about thrombolytic therapy for acute ischemic stroke among community residents in Yuzhong district,Chongqing,China.Methods We used multi-stage sampling to select 1500 households distributed in 3 Street of Yuzhong district,and then one member of each household answered the survey.A trained interviewer conducted a face to face survey about the questions regarding stroke warning signs and thrombolytic therapy in acute stroke.Results A total of 1101 participants completed the survey.Only 257 (23.3％,95％ CI 20.8-25.8) participants were aware of thrombolytic therapy for acute stroke and 59.9％ of them (154/257,95％ CI53.9-65.9) knew the time window of thrombolytic therapy.The rate of thrombolytic therapy awareness was higher among people with young age,well-educated,with higher household income with health insurance,and those who knew all 5 stroke warning signs.People aged ≥75 years had higher rate of awareness of the time window.Multivariate logistic regression analysis showed that age,education level,health insurance and knowledge of stroke warning signs were independently associated with the rate of thrombolytic therapy awareness.Conclusions In this population-based survey the community residents had poor awareness of thrombolytic therapy for acute ischemic stroke.It is necessary to improve the level of public knowledge about thrombolytic therapy for acute stroke by health education.

Objective Knowing the BRCA gene mutational condition of high risk triple negative breast cancer (TNBC) in Xinjiang Uygur Autonomous and acquiring the differences of clinical and pathologic characteristics between person with BRCA gene mutation and person without it by means of BRCA gene mutation testing for 30 cases of TNBC in Xinjiang Uygur Autonomous.Methods The objects of this study were 30 cases of high risk TNBC from Xinjiang.All the coded sequences of BRCA1/2 gene were amplified by means of extracting genomic DNA from peripheral venous blood.BRCA1/2 gene mutation analysis were prescreened through DHPLC.Then,the result was verified by DNA sequencing.The clinical and pathologic characteristics between person with BRCA gene mutation and person without it of 30 high risk TNBC cases were contrastively analysed.Results In all the 30 cases of BRCA gene mutation testing for TNBC in Xinjiang Uygur Autonomous,there were 5 cases of pathogenic mutations of BRCA gene (5/30,16.7 ％); 4 cases of BRCA 1 mutation (4/30,13.3 ％); 1 case of BRCA 2 mutation (1/30,3.3 ％); and there was no mutation to be found in 25 cases of BRCA gene of TNBC (25/30,83.3 ％).As compared with person without gene mutation,who with it had the characteristics of earlier of TNM,the difference was statistically significant (P =0.040).Conclusion Since the rate of BRCA1 gene mutation of high risk TNBC is higher.It is suggested that the BRCA gene of every patients with high risk TNBC should be tested.Comparing with person with BRCA gene mutation and person without it,there might have differences on clinical pathological characteristics features.Therefor,individualized treatment should be taken into consideration.

A bacterium strain BFHM2002 is isolated from Lake Donghu, Wuhan. BFHM2002 has advantages that it can produce melanin with a high rate and high yield in the absence of tyrosine. Induced by tyrosine, melanin yield can be dramatically increased. BFHM2002 may be identified as a new strain in Bacillus firmus, for melanin-production.

Objective To investigate the immunogenicity of immunodominant cytotoxic T lymphocyte epitope E749-57 of human papilloma virus (HPV) 16 oncoprotein E7 chaperoned by heat shock protein (HSP)110. Methods Mouse HSP110 gene was cloned into prokaryotic expression vector pQE-80L for the expression of HSP110 protein, which was purified using Ni-NTA column. SDS-PAGE and Western-blot were conducted to confirm the purified mHSP110 protein, which was subsequently incubated with E749-57 peptide under heat shock condition, and high-performance liquid chromatography (HPLC) was used to evaluate the binding efficiency of the recombinant protein and E749-57 peptide. Twenty mice were divided into 4 groups to be immunized with mHSP110 protein, E749-57 peptide, mHSP110-E749-57 complex and phosphate buffered saline (PBS),respectively. Two weeks after the last immunization, spleen cells were collected from the immunized mice and divided into 2 parts: one were stimulated by E749-57 peptide followed by the detection of CD8+ INF-γ+ T cells with flow cytometry; the other one were subjected to MTT analysis for the estimation of cell proliferation. The mHSP110-E749-57 complex was also used to immunize TC-1 tumor bearing mice to observe its anti-tumor effect.Results The full-length 2577 bp-sized mHSP110 gene was amplified from mouse liver cDNA and cloned into pQE-80L vector. Direct sequencing confirmed the correctness of the cloning. SDS-PAGE and Western-blot demonstrated the successful purification of mHSP110. HPLC assay showed that the purified mHSP110 protein could bind with E749-57 to form a relatively stable protein complex. The percentage of IFN-γ+ CD8+ T cells in and proliferation index of spleen cells from the complex-immunized mice were statistically higher than those from the other 3 groups of mice. Moreover, the complex could obviously inhibit the growth of TC-1 tumor in mice. Conclusion The mHSP110-E749-57 complex could enhance the generation of specific cytotoxic T lymphocytes and exert anti-tumor effects in mice.

OBJECTIVETo investigate the significance of detecting specific serum IgG antibodies in clinical diagnosis of SARS as well as affecting factors.

METHODSEnzyme-linked immunoassay kit for SARS coronavirus antibodies developed by HuaDa Biological Company was applied to detect specific serum IgG from SARS patients and the production of SARS specific antibodies among patients of different age groups, sex and with or without steroid treatment were statistically compared.

RESULTSOut of 121 patients studied, 71.1% were SARS specific IgG positive. Patients younger than 15 years, between 15 to 59 years, older than 59 years had positive rates of 60.0%, 70.2%, and 85.7%, respectively with no statistically significance (P=0.766); patients with or without steroid treatment showed positive rates of 70.6% and 72.4%, respectively (P=0.84); patients exhibiting either severe or light syndromes showed positive rates of 78.1% and 67.4%, respectively (P=0.493); both male and female patients showed the same positive rate of 71.1%.

CONCLUSIONThe sensitivity of the SARS specific IgG kit utilized needs to be further improved. The production of SARS IgG is not notably correlated with sex, age, seriousness of symptoms, and steroid treatment.

Adolescent ; Adult ; Aged ; Antibodies, Viral ; blood ; Child ; Female ; Humans ; Immunoglobulin G ; immunology ; Male ; Middle Aged ; SARS Virus ; immunology ; Sensitivity and Specificity ; Severe Acute Respiratory Syndrome ; diagnosis ; immunology

Objective To investigate the efficacy of the bioactive artificial vertebrae of a nano- hydroapatite crystals and polyamide 66 composite(n-HA/PA66)to restore the height and architecture of thoracolumbar burst fracture.Methods From December 2003 to February 2006,38 patients(29 males and 9 females)with a mean age of 35.6 years(17-63 years)were treated surgically through anterior ap- proach for decompression and implanted with the bioactive artificial vertebrae of n-HA/PA66 composite to reconstruct the structure of the thoracolumbar burst fractured vertebra.Results All the patients were successfuly followed-up for an average of 8 months,ranging from 6 to 21 months.The bioaetive artificial vertebrac of n-HA/PA66 composite were fused with the receptor bone 3-4 months after operation.The neu- rological function of the patients was restored partially or completely.The thoracolumbar spine was stable during physical examination and the height of thoraeolumbar burst fractured vertebrae that had been restored did not changa during the follow-up.Conclusions Our results show the bioaetive artificial vertebrae of n-HA/PA66 can restore the height and structure of thoracolumbar burst fractured vertebrae and reconstruct the structure of the tboraeolumbar vertebrae effectively,indicating that the bioaetive artificial vertebrae of n- HA/PA66 can be used extensively in clinical spinal surgery.

PURPOSE: The aim of this study is to further understand the status of BRCA1 and BRCA2 mutation among Chinese high-risk breast cancer patients in multiple-ethnic regions of China. METHODS: A total of 79 blood samples of high-risk breast cancer patients from Xinjiang Uyghur autonomous region were analyzed by PCR-DHPLC sequencing analysis. RESULTS: Analysis with full length of the two genes identified a total of 6 deleterious mutations (2073delA, 2394C-T [Q759X] and IVS16+1G>A in BRCA1; 1627A-T [K467X], 6873delCTCC and 9481delA in BRCA2) in this cohort. The prevalence of BRCA1/2 germline mutation was about 7.6% (6/79) in the Xinjiang multiple ethnic region of China. Among them, 3 novel deleterious mutations, 2073delA in BRCA1 (Han ethnic Chinese) and BRCA2 variants 6873delCTCC and 9481delA (both are Kazakh ethnic Chinese), were identified and they had never been reported in breast cancer information core (BIC) database before. 2394C-T (Q759X) and IVS16+1G>A, in BRCA1 and BRCA2 variants 1627A-T were previously reported in other populations but not Chinese. Among 6 of the BRCA-related tumors, three BRCA1- and one BRCA2-associated tumors were in triple negative (estrogen receptor, progesterone receptor, and HER2 negative expressed) status and exhibited a high tumor grade. So far none of these 6 deleterious mutations were reported in ethnic Han Chinese. CONCLUSION: BRCA germline mutation in Chinese multiple ethnicity region may exhibit different genotypes compared to ethnic Han Chinese in other regions. These differences may arise from interaction of genetic background and environmental factors.
Asian Continental Ancestry Group ; Breast ; Breast Neoplasms ; China ; Cohort Studies ; Ethnic Groups ; Female ; Genetic Variation ; Genotype ; Germ-Line Mutation ; Humans ; Prevalence ; Receptors, Progesterone

OBJECTIVETo investigate the HCV genotypes distribution in northern and southern cities in China and the difference between patients infected with HCV by transfusion and non-transfusion routes.

METHODSThe HCV genotypes of the patients with chronic hepatitis C from 9 cities belonging to different regions were genotyped by the PCR products of 5 prime untranslated region NTR digested with restriction endonucleases, and the HCV genotypes distribution among different cities or between the patients infected with HCV through transfusion and other routes was analyzed.

RESULTSThe HCV genotypes of 214 in 219 cases were determined; 197 patients were infected with monogenotype HCV. The major epidemic genotypes of HCV isolates in China were 1b (76.64%) and 2a (18.22%), but 5.14% of patients were infected with HCV belonging to genotype 3b and this was the first report that there is genotype 4a in China. The HCV genotype distribution was not different in northern and southern areas, but was significantly different between patients infected with HCV through transfusion and non-transfusion routes (P=0.036). In patients infected trough transfusion, the rates of monogenotype HCV infection and genotype 1b were 93.88% and 76.87%, respectively, which were higher than those (86.57% and 58.21%) in the patients infected with HCV through non-transfusion routes. The rate of patient infected with mixed genotype HCV strains in non-transfusion group was 13.43%, which was higher than that (6.12%) of patients in transfusion group.

CONCLUSIONThe HCV genotype distribution in northern and southern regions were similar, but was significantly different between the patients infected through transfusion and other routes.

5' Untranslated Regions ; Adolescent ; Adult ; Aged ; China ; Female ; Genotype ; Hepacivirus ; classification ; genetics ; isolation & purification ; Hepatitis C, Chronic ; etiology ; genetics ; transmission ; Humans ; Male ; Middle Aged ; Transfusion Reaction

BACKGROUNDThe capsule associated protein 10 gene (cap10) is indispensible for the formation of the polysaccharide capsule, and is important in maintaining virulence of the Cryptococcus (C.) neoformans. In this study, we aimed to construct an short hairpin RNA (shRNA) expression vector targeting C. neoformans cap10 gene expression and confirm its biologic relevance.

METHODSA pair of oligonucleotides targeting the cap10 cDNA sequence was designed and synthesized. It was cloned into the plasmid psilencer4.1-CMV neo to construct an eukaryotic shRNA expression vector. The vector was transfected into C. neoformans cells using the LiAc method. The expression of cap10 was assessed by real-time fluorescence quantitative PCR. Groups of C. neoformans cells were incubated with murine macrophage-like J774A.1 cells, and the phagocytic indexes and ratios were determined by the microscopic observation method.

RESULTSThe expression of cap10 in C. neoformans cells transfected with ps4.1 neo-cap10 ((175,535.00 ± 47,004.00) copies/µl) was lower than that of cells transfected with the empty vector ((512,698.89 ± 32,318.02) copies/µl) and mock transfected cells ((562,931.66 ± 65,928.41) copies/µl). The average phagocytic ratio and phagocytic index of J774A.1 cells following incubation with C. neoformans were higher for cells transfected with ps4.1 neo-cap10 (0.21 ± 0.02, (19.06 ± 1.66)%) than for the control experimental group (0.08 ± 0.02, (6.57 ± 1.23)%) and the blank experimental group ((0.07 ± 0.01), (5.89 ± 1.07)%) (P < 0.05).

CONCLUSIONSThe cap10 shRNA vector was successfully prepared and transfected into C. neoformans cells. The effect of RNA interference on the expression of the C. neoformans cap10 gene is effective, and it can induce phagocytosis of C. neoformans.

Animals ; Cell Line ; Cryptococcus neoformans ; genetics ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Genetic Vectors ; genetics ; Phagocytosis ; Plasmids ; genetics ; Polymerase Chain Reaction ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection