Child ; Female ; Humans ; Polyendocrinopathies, Autoimmune ; classification ; diagnosis ; therapy ; Treatment Outcome

Comparison studies of recombinant prourokinase(pro-UK) in various host cells,and expression vectors were carried out. Expression levels of vectors constructed in this study in different cell lines were compared. Mammalian cells expressing pro-UK were established. The levels of pro-UK expression in recombinant Namalwa, Vero and Sp2/0 cells are 200,12.5 and 50 IU/( 106 cells 24h). pro-UK purities separated by immunoaffinity chromatograph are above 90 %. Immunoabsorbent assay showed the ratio of pro-UK in CHO cells is lower than that from Vero and Namalwa cells. This study provide new host cells for pro-UK production.

OBJECTIVETo establish immortalized cell line from the urothelium of the urinary bladder and identify the characteristics of the cell line.

METHODSHuman papillomavirus 16 (HPV-16) plasmid was used to transfect urothelium of infant urinary bladder in vitro with the help of Fugene-6, and this plasmid contained E6 and E7 genes of HPV-16. We also identified the existence of HPV-16 E6 and E7 genes and the biological characteristics of the cell line by PCR, immunohistochemistry, and the biology identification.

RESULTSBLTR-4 cell line, produced from the transfection of HPV-16K plasmid, was a cell line from urothelium with the expression of HPV-16 E6 and E7 genes. It had been cultured more than 70 passages, and the characteristics of growth was similar to the immortalized cell line as reported.

CONCLUSIONSBLTR-4 cell line is an immortalized cell line from urothelium of the urinary bladder, which contains HPV-16 E6 and E7 genes. BLTR-4 cell line is a good experimental model to investigate the relationship of the infection of high risk HPV and transitional cell carcinoma (TCC) in vitro.

Cell Line, Transformed ; Humans ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Papillomavirus Infections ; virology ; Plasmids ; genetics ; Repressor Proteins ; genetics ; Transcription, Genetic ; Transfection ; Tumor Virus Infections ; virology ; Urinary Bladder ; cytology ; Urinary Bladder Neoplasms ; virology

OBJECTIVETo isolate, identify and culture human spermatogonial stem cells (SSC) and then obtain purified and enriched human SSCs for research and application.

METHODSWe detected the expression of CD90 in the human testis using the immunofluorescence technique and isolated human testicular spermatogenic cells by two-step enzymatic digestion, followed by differential plating and magnetic-activated cell sorting (MACS) with CD90 as an SSC marker. Then we identified the isolated CD90-positive spermatogenic cells by RT-PCR and immunocytochemistry, and meanwhile cocultured them with Sertoli cells in SG medium in vitro.

RESULTSThe isolated CD90-positive cells showed a relatively homogeneous characteristic in size and morphology and expressed the genes specific for human SSCs, with high expressions (90.5%) of GFRA1, GPR125, and UCHL1. After coculture with Sertoli cells in the SG medium for 2 weeks, the isolated CD90-positive cells maintained a good activity.

CONCLUSIONCD90 can be regarded as a speci- fic marker for human SSCs and used to obtain highly enriched human SSCs by differential plating and MACS. Furthermore, the isolated human SSCs can be cultured in SG medium in vitro.

Adult Stem Cells ; cytology ; Biomarkers ; metabolism ; Cell Separation ; methods ; Cell Shape ; Cell Size ; Coculture Techniques ; Glial Cell Line-Derived Neurotrophic Factor Receptors ; metabolism ; Humans ; Immunohistochemistry ; Male ; Receptors, G-Protein-Coupled ; metabolism ; Sertoli Cells ; Spermatogonia ; cytology ; Testis ; metabolism ; Thy-1 Antigens ; isolation & purification ; metabolism ; Ubiquitin Thiolesterase ; metabolism

Objective To observe expression of neuron specific enolase (NSE) gene in the brain of rats induced by iodine excess. Methods One hundred and fifty one-month weaning Wistar rats were divided into ten groups according to 5 × 2 factorial experiment. Rats were fed with normal feedstuff and water of a series of iodine concentration by adding potassium iodide respectively: norrmal iodine (NI), five-fold high iodine (5HI), ten-fold high iodine(10HI), fifty-fold high iodine(50HI) and one hundred-fold iodine(100HI). After these rats were fed for three or six months, rat serum thyroid hormones were measured by radioimmunoassay including TT4, TT3, FT4, FT3, rT3 and the mRNA level of NSE in rat brain tissue was studied using RT-PCR technique. Results The levels of serum TT4 and TT3 were significantly different in five iodine level groups(F values were 18.867,27.287, both P < 0.01). The interaction between time and iodine level in TT4 was significant in our study(F values were 2.486, P < 0.05). The levels of TT4 and TT3 of 100 HI group at third and sixth month were lower than those of NI, 5HI, 10HI, 50HI groups in the same period (all P < 0.01). The levels of serum FT4, FT3 and rT3 were significantly different at different time(F values were 4.968,27.046,59.776 respectively, P < 0.05 or < 0.01) and in different iodine level groups(F values were 33.058,28.420,17.482 respectively, all P < 0.01). Moreover, the interaction between time and iodine level in FT3 and rT3 was significant in our study(F values were 6.894,5.233 respectively, both P < 0.01). FT4, FT3 and rT3 in 100HI group were lower than that of other iodine dosage groups at the same time (P < 0.05 or < 0.01). The levels of NSE mRNA in brain tissue was significantly different in five iodine level groups (F values were 29.006, P < 0.05). The levels of mRNA NSE of 100HI group in both three and six months (0.61 ± 0.19,0.61 ± 0.22) were all lower than that of any other groups[NI(0.73±0.13 and 0.72 ±0.26), 5HI (0.72 ± 0.15 and 0.72±0.16), 10HI (0.73 ±0.32 and 0.70±0.13), 50HI(0.71±0.18 and 0.69±0.31), all P < 0.05]. The results of correlation analyses show that the levels of serum FT3 and FT4 had correlations with the levels of NSE mRNA (P < 0.05) both in three and six months(r values were 0.987, 0.969 in three month, and 0.890, 0.910 in six month respectively). Conclusions The expression of NSE gene can tolerant the excess of iodine to a certain extent. Exposure to heavy excess iodine(100HI) can decrease the mRNA level of NSE gene. FT4 and FT3 may both have important roles on the regulation of NSE mRNA induced by excess iodine.

AIM:To investigate the synergistic effect of imperatorin on enhancing the anti -tumor effect of TNF-related apoptosis-inducing ligand(TRAIL)on breast cancer and the mechanisms.METHODS:T-47D and MCF-7 breast cancer cells were divided into control group ,imperatorin group,TRAIL group,imperatorin+TRAIL group and imperatorin+TRAIL+death receptor 5(DR5)siRNA group.The viability of T-47D and MCF-7 cells was measured by MTT assay. The apoptosis and mitochondrial membrane potential in T-47D cells were analyzed by flow cytometry.Western blot and flow cytometry analysis were performed to evaluate the expression of DR 5 on T-47D cell surface and the activation of caspase-8 and caspase-3.RESULTS:Imperatorin significantly enhanced the inhibition of cell viability induced by TRAIL of T -47D and MCF-7 cells,and significantly increased the apoptosis of T-47D cells induced by TRAIL.Imperatorin treatment ob-viously induced upregulation of DR5 expression and production of reactive oxygen species in the T-47D cells.In addition,imperatorin enhanced the TRAIL-induced damage of mitochondrial membrane potential and activation of caspase -8 and caspase-3.CONCLUSION:Imperatorin enhances the anti-tumor effect of TRAIL on breast cancer via upregulating the ex-pression of DR5.

Objective To study on epidemiologic characteristics of human plague cases in Sanjiangyuan Area,and provide theoretical basis to work out the preventive measures.Methods Based upon the epidemiology information from the human plague case data bank of Qinghai Province,human plague data were analyzed retrospectively in Sanjiangyun Area by sorting,verifying and summing up of these data,including some of case file and monitoring data.Results Except for 12 years in the period of 1960 to 2006,there were human plague cases happened every year.The morbidity occurred mainly in 12 counties of 4 states,including Yushu,Guolou,Huangnan and Hainan,and Tanggula Town of Geermu City,a total of 85 human plague episodes were occurred,resulting 238 onsets,134 deaths,and a matality rate of 56.30%.The sources of infection were respectively Himalayan mormot 27.31%(65/238),artiodactyls 14.71%(32/238),carnivora 2.10%(5/238),Lagomorpha 0.42%(1/238),the pneumonic plague patient 49.16%(117/238),and biting of flea 6.30%(15/238).The prevalent season was from May all the way to November,the peak-months were August and September.After October,the sheep as the source of infection initiating human being plague accounted for 23.53%.Among the clinical types,the most prevalent type was pneumonic type(61.34%),and the rest,glandular type(17.23%),septic type(16.81%)and other types(4.62%),but the first plague case in each epidemic was mainly the glandular plague.Conclusions In recent years,the tendency of human plague prevalence increases in Sanjiangyuan Area,it is urgent to improve and adjust the prevention and treatment measures in time.

OBJECTIVETo describe the prevalence of metabolic syndrome (MS) in Jiangsu province, using IDF 2005 and ATP III 2005 definition.

METHODSMultistage cluster sampling method was used and the participants were interviewed by trained health workers under a structured questionnaire. The number of research subjects was 5888. Fasting plasma glucose (FPG), blood lipids, body mass index, blood pressure (BP) of all sample population (5888 subjects) were measured.

RESULTSThe prevalence of MS was 17.48% by IDF 2005 definition and 21.95% by ATP lII 2005 definition. Women had higher prevalence than men (P < 0.01). The prevalence of MS increased with age. Abrupt increase of the prevalence started at age of 50 in women. The prevalence varied by education, marital status, occupation and income. People from the urban and the south had a higher prevalence than from the rural and from the northern part of the country. The most common combination of MS individual components were 1) central obesity, dislipidemia and high BP; 2) central obesity, dislipidemia, high BP and high FPG.

CONCLUSIONThe prevalence of MS was high in Jiangsu province. Clustering of MS components was common. It is necessary to discuss the cut-off points of central obesity for the waist circumference diagnostic criteria of MS in Chinese population. The importance of prevention of MS should be strengthened both by health professionals and the government.

China ; Female ; Humans ; Male ; Metabolic Syndrome ; epidemiology ; Middle Aged ; Obesity, Abdominal ; epidemiology ; Prevalence ; Waist Circumference

OBJECTIVETo compare the significance of the application in Jiangsu population using the diagnostic criteria for metabolic syndrome(MS) proposed by the IDF in 2005, ATP III in 2005 and CDS in 2004.

METHODSBased on the populations in Jiangsu province from a project of Multiple Metabolic Disorders and MS, the study was conducted including 5888 cases, with data of plasma glucose, lipid profile, blood pressure, serum insulin etc. MS was diagnosed and compared according to these three definitions respectively.

RESULTSThe age-adjusted MS prevalence rates were 17.48%, 21.95% and 9.59% according to the IDF(2005), ATP III (2005) and CDS (2004) respectively. The agreement in the diagnosis of MS using ATP III (2005) and CDS(2004) definitions was 85.11%, and the agreement in the diagnosis of MS using IDF(2005) and CDS definitions was 87.35%. The agreement in the diagnosis of MS using IDF (2005) and ATP III (2005) definitions was 95.14%. The MS subjects diagnosed by the ATP III (2005) was 1.26 higher than subjects diagnosed by the IDF(2005) definition. The ratios of prevalence rates of high waist circumference(WC), MS_IDF (2005) and MS_ATP III (2005) was 2.17, 1.99 and 1.54 in sex ratio (woman to man).

CONCLUSIONThe agreement in the diagnosis of MS using the IDF(2005) and ATP III (2005) definition was higher than using CDS(2004) and other two definitions. For diagnosing MS, the cut off of WC in IDF(2005) and ATP III (2005) seemed not appropriate and the diagnostic criteria used for ATP III (2005) (waist circumference of man 85 cm, woman 80 cm) could identify more MS.

Blood Glucose ; Blood Pressure ; China ; Humans ; Insulin ; blood ; Lipids ; blood ; Metabolic Syndrome ; diagnosis ; Reference Values ; Waist Circumference

OBJECTIVETo observe the effect of overdose iodine on the expression of CCK gene in brains of rats and identify the possible mechanisms.

METHODSOne-month weaning Wistar rats were randomly divided into five groups which were fed with normal feedstuff and water supplemented with different concentrations of potassium iodide, named A group (iodine ration was about 6.15 microg per day), B group (iodine ration was about 30.75 microg per day), C group (iodine ration was about 61.5 microg per day), D group (iodine ration was about 307.5 microg per day) and E group (iodine ration was about 615 microg per day). Rats were sacrificed after being fed for three or six months. Then serum thyroid hormones were measured by radioimmunoassay and the mRNA level of CCK gene was studied by using RT-PCR technique.

RESULTSAt the end of three months, the values of thyroid hormones in E group [TT4 (45.2 +/- 13.7) nmol/L, TI'3 (0.65 +/- 0.20) nmol/L, FT3 (0.93 +/- 0.45) pmol/L, FT4 (7.07 +/- 2.43) pmol/L, rT3 (0.15 +/- 0.04) nmol/L] were all lower than those in A group [TT4 (76.0 +/- 18.8) nmol/L, TT3 (1.34 +/- 0.41) nmol/L, FT3 (2.45 +/- 0.62) pmol/L, FT4 (15.12 +/- 3.40) pmol/L, rT3 (0.24 +/- 0.04) nmol/L]. There were significant differences between E group and A group on the levels of serum TH (F values are 14.68, 16.03, 21.16, 20.25, 13.52 respectively, P < 0.01); FT3 levels in C and D groups were significantly decreased as compared to A and B groups (F = 21.16, P < 0.05). rT3 level in D group was significantly decreased compared with A,B and C groups (F = 13.52, P < 0.05). At the end of six months, the levels of serum TH in E group (TT4 (51.84 +/- 15.83) nmol/L, TT3 (0.77 +/- 0.22) nmol/L, FT4 (6.88 +/- 2.23) pmol/L, FT3 (0.74 +/- 0.28) pmol/L, rT3 (0.14 +/- 0.03) nmol/L) were lower than those in any other groups (F values were 6.05, 12.22, 11.25, 13.42, 5.89 respectively, P < 0.05). At the end of both three and six months, the mRNA levels of CCK gene in E group were lower than any other groups (F values were 4.04, 3.95 respectively, P < 0.01). The results of correlation analysis showed that serum FT4 had linear correlation with levels of CCK mRNA (r values were 0.990, 0.948 respectively; P < 0.05); However serum FT3 had no linear correlation with the levels of CCK mRNA (r values are 0.970, 0.932 respectively).

CONCLUSIONSExposure to overdose of iodine (iodine ration was 100-fold higher than that of A group) could decrease the mRNA level of CCK gene. Compared with FT3, FT4 might have more important role on the regulation of CCK mRNA induced by excess of iodine.

Animals ; Brain ; metabolism ; Cholecystokinin ; biosynthesis ; genetics ; Drug Overdose ; Female ; Food, Formulated ; Gene Expression ; Hyperphagia ; Iodine ; toxicity ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Thyroid Hormones ; blood ; Thyrotropin ; blood ; Thyroxine ; blood ; Triiodothyronine ; blood