Objective To study isolation and identification and culture of rat type A spermatogonial cells in vitro.Methods Percoll discontinue density gradient centrifugation combined with different speeds of different cells adhering to dish was used to purify the type A spermatogonial cells.The c-kit and TERT special antibodies were used to identify the type A spermatogonial cells.The purified cells were cultured in vitro. Results 0.614?10~6 cells per testis finally were obtained and the percentage of viable cells was 92.1% by trypan blue dye exclusion test.The percentage of type A spermatogonial cells expressing c-kit and TERT were 91.7?1.2% and 90.8?1.0% respectively.Type A spermatogonial cells could proliferate and self-renew in the DMEM containing 10% NBS.Conclusion Percoll discontinue density gradient centrifugation combined with different speeds of different cells adhering to dish is an efficiency method for isolation of rat type A spermatogonial cells.The purified cells are type A spermatogonial cells by identification of the immunohistochemistry of c-kit and TERT antibodies.Type A spermatogonial cells can proliferate and self-renew in vitro.

Objective:To assess human leucocyte antigen(HLA)-DRB1 allele polymorphism and analyze the association between DRB1 allele polymorphism and clinical classification of ulcerative colitis.Methods:60 patients were investigated for DRB1 gene by DNA microarray.The results were compared with those from healthy subjects.Results:The frequency of DR2 and DRB1*15 in UC patients were 45% and 41.7% respectively,which were significantly higher than those(23.3% and 21.7%)in controls,the odds ratio being 2.688 and 2.582 respectively(P

Objective:To investigate the distribution and expression of the cyclooxygenase-2(COX-2)in lung tissues and the effect of its inhibition in bleomycin(BLM)-induced pulmonary fibrosis in rats.Methods:72 male SD rats were randomly divided into three groups:the control group,model group and celecoxib group.Immunohistochemical method was used to detect the distribution of COX-2 in the stage of acute pulmonary alveolitis.HE and Masson staining methods were used to evaculate the degree of alveolitis and pulmonary fibrosis.Results:(1)In early stage of pulmonary fibrosis except in the control group,COX-2 expression was found in small bronchial epithelial cells in the other two groups,especially the model group.(2)Compared with the control group,the other two groups had alveolitis(P

BACKGROUND: Having breakfast everyday is a behavior promoting health recommended WHO, while it is easy to be ignored.OBJECTIVE: To investigate the breakfast habits of pupils in Beijing and Shanghai so as to provide a scientific basis for further studying the problems in having breakfast among the pupils, drafting and developing "nutritious breakfast program for students".DESIGN: Stratified and cluster sampling and investigation study were used based on the pupils.SETTING: Institute of health for child of two universities.SUBJECTS: A total of 8688 six to eleven years old Beijing and Shanghai pupils were involved from March 2002 to June 2002. Among them 4692 pupils came from Beijing. There were 4637 valid questionnaires, 2386 of which were from male pupils and 2251 from female pupiIs. Among the total subjects, there were 4300 Shanghai pupils, and 4051 questionnaires were valid, of which 2031 were from the male pupils and 2020 from female students.METHODS: A self-administered questionnaire survey was conducted. The content of the questionnaire included the general conditions of the family, dietary preferences, food for breakfast, consumption of the milk food and so on.RESULTS: The results showed that about 25.8% boys and 21.6% girls in Beijing did not eat breakfast regularly; similarly, the percentage was about 11.4% for boys and 13.3% for girls in Shanghai. The reasons for not having breakfast included the following: They had no appetite, or had no enough time for it, or the parents were too busy to prepare in the mornings. For those of them who eat breakfasts, the nutrients of their breakfast usually were not adequate. 55% of the students had their breakfast prepared by their mothers. As for the eating sites, students mainly ate at home; the next was having it on the way to school or eating at the roadside luncheonettes. The cost of breakfast was 2 - 3 yuan. The breakfast behaviors of the pupils were closely related with the behaviors of their mothers( r = 0. 4333, P < 0.01 ).CONCLUSION: It is suggested that quality of breakfast of the pupils need to be improved. The behaviors of the parents influence their children in breakfast habits.

Objective To inspect the correlation of synthetic effect criteria to sensitivity,reliability of late stage lung cancer curative effect verdict and median survival time. Method Using size change of tumor,symptom change related to tumor,Karnofsky score,body weight etc.as synthetic effect criteria and nstituting clinical synthetic effect criteria,to assess the curative effect of late stage lung cancer patients who received different treatment in department of tumor of Xiyuan Hospital between March 1997 and March 2000, to analysis medium survival time, and being compared with RR of tumor.Result 52 patients entered this experiment, in which 42 patients were assessable (follow up), 13 patients were effective(30.95％ ),17 were patients stable(40.48％ ),12 patients deteriorated (28.57％ ).The result of tumor objective remission rate: CR is 0,PR is 16.67％ (7 patients),NC is 64.29％ (27 patients) and PD is 19.05％ (8 patients).There was apparent difference between two effect criteria(P< 0.05).The mean survival time of this group is (9.3± 1.6)months,median survival time is 10.5 months(2～ 8months),survival rate of one year is 33.33％ (14 patients).In the patients who live longer than median survival time, most of them survive associated with tumor existence who are in NC of tumor objective remission rate, but also in the effective column of synthetic effect criteria. Compared with tumor objective remission rate,the synthetic effect criteria have more apparent correlation to survival time. Conclusion In the assessment of late stage lung cancer,synthetic effect criteria has better sensitivity and reliability,and emphasize survival quality,at same time it reflect the reaction of tumor and host to treatment,it reflect prognosis of tumor preferably and have better correlation to the survival time. It can be used as clinical effect criteria of late stage lung cancer after further consummation.

BACKGROUND: Deficit of nivalenol (NIV) and selenium (Se) is related with kashin-beck disease (KBD) to certain extent. Hyaluronic acid (HA) metabolism affects directly the polymerization of proteoglycans (PG) and normal structure and function of cartilage. The integration with HA receptor on surface of cartilage tissue is the key link in HA metabolism. Being the main receptor of HA on chondrocytic membrane, CD44 expression impacts directly HA metabolism, further affects cartilage matrix metabolism, which is extremely important to maintaining the structure and function of cartilage matrix.OBJECTIVE: To probe into the injury and protection of related etiology of KBD to target tissue cells and the mechanism on degenerative necrosis of chondrocytes.DESIGN: Randomized controlled observation was designed.SETTING: Department of Genetics and Molecular-Biology of Medical College of Xi' an Jiaotong University.MATERIALS: The experiment was performed in Key Laboratory Room on Ministry of Education associated with Environment and Disease of Xian Jiaotong University from October 2002 to July 2004. One New Zealand pedigree young rabbit aged 30 days was employed and its humerus, femurs and tibia were cut out in surgery.METHODS: With cell culture, the model of bone tissue was reconstructed in vitro, in which, NIV of various concentrations, KBD suspicious infectious agent and Se, the protective factor were added. HA receptor CD44 on chondrocytic membrane and soluble CD44 in cell culture solution were determined.MAIN OUTCOME MEASURES: ① Microscopic observation of adhesion molecule CD44 on chondrocytic surface. ② Soluble CD44 in chongrocytic culture solution.RESULTS: ① Microscopic observation of adhesion molecule CD44 on chondrocytic surface: CD expression in chondrocytic membrane was decreased with increasing of NIV concentration and it was in tendency of increasing with Se added. ② Soluble CD44 in chongrocytic culture solution:The concentration of soluble CD44 in cell culture solution was decreased gradually following the increased concentration of NIV, but it was increased in high concentration group and such tendency did not alter when Se added. Except blank control and Se control, significant difference was presented among groups (P ＜ 0.05).CONCLUSION: NIV disturbs adhesion molecule CD44 expression on chondrocytic surface and further induces metabolic disturbance of cartilage extracellular matrix. Se supplementation can resist the injury of NIV to chondrocytes, but its action is limited.

Objective To investigate the gene expressions of IL-1 and TNF-? in bronchoalveolar lavage fluid(BALF) and lung tissues in mice with pulmonary fibrosis(PF) at various periods and to study the effects of cytokines on pulmonary fibrosis.Methods Forty male mice were randomly divided into two groups:experimental group(n=30) and control group(n=10),and poured respectively by bleomycin solutions and saline and killed on the 3rd,7th,14th,28th and 56th day.RT-PCR method was used to analyse the mRNA expressions of IL-1 and TNF-?.Results ① The expressions of IL-1 mRNA and TNF-? mRNA in BALF in experimental group were higher than those in control group(P

Objective To investigate the effects of capparis spinosa total alkaloid on type collagen (ColⅣ - ),Ⅳmatrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1) and plasminogen activator inhibitor-1 (PAI-1) in bleomycin-induced systemic sclerosis (SSc) mice; To explore the effective mechanism of capparis spinosa total alkaloid on fibrosis of SSc. Methods Totally 90 BALB/c mice were randomly divided into control group, model group, penicillamine group and capparis spinosa total alkaloid low-, medium- and high-dose group. Mice models with SSc were established by repeated local injections of bleomycin in mice back, except for the control group. Mice in medication groups received external application with capparis spinosa total alkaloid cream;mice in penicillamine group were given penicillamine for gavage; mice in the control and model group received external application without substance, one time a day, for 60 days. The contents of MMP-9, TIMP-1 and PAI-1 in serum and Col- in skin tissue were dⅣ etected respectively by ELISA after the last medication. Results Compared with the model group, the levels of MMP-9 and ratio of MMP-9/TIMP-1 markedly increased and the levels of Col-Ⅳand TIMP-1 markedly decreased in medium and high- dose of capparis spinosa total alkaloid group (P<0.05, P<0.01). But the level of PAI-1 was not influenced (P>0.05). Conclusion Capparis spinosa total alkaloid is effective in treating fibrosis of SSc by adjusting imbalance of MMP-9/TIMP-1 and decreasing expression of Col-Ⅳ.

Objective To investigate the effects of capparis spinosa total alkaloid on the pathological changes and the type Ⅲ collagen(COL?Ⅲ)expression in systemic sclerosis(SSc)mice. Methods Mice models with SSc were established by repeated local injection of bleomycin in BALB/c mice back. After administration of capparis spinosa total alkaloid ,the pathological changes of skin and lung tissue were observed ,and the COL?Ⅲ expression was detected by ELISA. Results Compared with the model group,the inflammation and fibrosis of skin and lung tissue were improved,and the level of COL?Ⅲ was markedly reduced by treatment of high dose capparis spinosa total alkaloid(P<0.05). Conclusion Cap?paris spinosa total alkaloid is effective in treating fibrosis of SSc.

Objective To investigate the expressions of matrix metalloproteinases(MMPs) in Kashin-Beck disease(KBD) cartilage as well as in a KBD rat model of T-2 toxin poisoning under selenium deficient conditions, and to investigate the effect of T-2 toxin on MMP-13 expression in human chondrocytes in vitro in order to determine a possible mechanism underlying KBD. Methods Samples of articular cartilage were divided into 2 groups:controls(samples from 5 normal children, traffic accident or operation), and KBD(samples from 5 children with KBD, auctopsy). Thirty-two Sprague-Dawley rats were divided into two groups by body weight using random number table: normal diet group(n = 16) and selenium-deficient diet group(n=16). The selenium level in normal diet was 101.500μg/kg, and in selenium-deficient diet was 1.118μg/kg. Rats were fed for 4 weeks with selenium-deficient or normal diet, respectively. After successful build up of the low selenium rat model, normal diet group was then subdivided into 2 sub-groups: normal group(n = 8) and normal diet plus low T-2 toxin group(n = 8);and selenium-deficient diet group was also subdivided into 2 sub-groups: selenium-deficient group ( n = 8 ) and selenium-deficient diet plus T-2 toxin group ( n = 8 ) . T-2 toxin of 100 μg·kg-1·d-1 was administered by intragastric administration for 30 days. Then the rats were sacrificed, and their knee joints were processed for histopathological evaluation. MMP-1 and MMP-13 locations in cartilages were performed by inmmunohistochemistry. Human chondrocytes C28/I2 were cultured in vitro. The experiment was divided into 4 groups: empty vector plasmid group, MMP-13 promoter plasmid group, MMP-13 promoter plasmid plus 20 μg/L T-2 toxin group and MMP-13 promoter plasmid plus 40 μg/L T-2 toxin group. MMP-13-luciferase reporter plasmid and vector plasmid were transiently transfected into C28/I2 cells for 24 hours, and then treated with 20 - 40 μg/L T-2 toxin for 24 hours. Transactivation of human MMP-13 promoter was analyzed using luciferase reporter constructs containing sequences spanning-1602 to+20 bp in C28/I2 chondrocytes. Results The percentages of chondrocytes staining for MMP-1 in the superficial and middle zones of KBD samples [(29.73 ± 10.12)%, (28.27 ± 0.91)%] were significantly higher than those of controls[(2.47 ± 0.11)%, (0.00 ± 0.00)%, all P < 0.05]. The percentages of chondrocytes staining for MMP-13 in the superficial and middle zones of KBD samples [(13.21 ± 4.32)%, (41.85 ± 6.32)%] were significantly higher than those of controls[(5.72 ± 0.31)%, (0.00 ± 0.00)%, all P<0.05]. The percentages of chondrocytes staining for MMP-13 in the superficial and middle zones of rats fed with selenium-deficient diet plus T-2 toxin group[(13.21 ± 4.32)%, (61.85 ± 8.68)%] were significantly higher than those of the normal and selenium-deficient groups[(2.43 ± 0.22)%, (5.89 ± 0.69)%, (3.03 ± 0.29)%, (25.99 ± 0.57)%, all P < 0.05]. Moreover, T-2 toxin activated the MMP-13 promoter detected with luciferase reporter assays in C28/I2 cells. The luciferase activities in MMP-13 promoter plasmid plus 20 μg/L T-2 toxin group and MMP-13 promoter plasmid plus 40μg/L T-2 toxin group(0.082 78 ± 0.008 40, 0.103 35 ± 0.013 19) were significantly higher than those in empty vector plasmid group and MMP-13 promoter plasmid group(0.024 19 ± 0.000 96, 0.040 32 ± 0.003 56, all P < 0.05). Conclusions These data suggest that T-2 toxin induces cartilage matrix degradation through up-regulation of MMP-13 promoter expression. Increased MMPs staining intensity in KBD cartilage and the rat KBD model of T-2 toxin poisoning under selenium deficient conditions suggest that matrix degradation appear to be driven by MMPs activity.