Objective To detect the osteopontin(OPN)expression in renal tissue of rats with fluorosis and low calcium diet,and study the role of OPN in renal injury of fluorosis.Methods Forty-eight aged 1 month Wistar rats,80-120 g,were randomly divided into 4 groups by 2×2 factorial design(the number of female and male in each group was equal):the control group,high-flluoride group,low-calcium group and low-calcium with high-fluoride group.All rats of the fluorosis groups were fed with feed containing corn exposed to coal-burning from endemic fluorosis areas with high fluoride(100 mg/kg,corn),the other two groups were fed with feed containing coru from nonendemic fluorosis areas(fluoride 5 mg/kg,corn).After 16 weeks,the rats were killed.The change of teeth was examined,and the incidence rate of dental fluorosis was calculated.The expressions of both protein and mRNA of OPN in rat renal tissue were determined by RT-PCR and immunohistochemistry after four-month experimentation.Results The growth of teeth was very well in the control group and the low-calcium group.The two high-fluoride groups showed evident dental fluorosis(100%).The results of immunohistochemistry showed that the OPN protein was localized in renal tubule cytoplasm.The OPN-positive cells from renal tissue were lightly and scatteredly stained in control and low-calcium groups.The OPN-positive cells had deeper color in high-fluoride group and low-calcium with high-fluoride group,widely distributed in the renal tubular epithelial cells.The protein expression of OPN in the two groups exposed to fluoride(168.64±13.21,169.26±8.92)was significantly higher than those of the corresponding control group(145.78±10.26,all P<0.01)and low-calcium group(149.60±16.84,all P<0.01).The mRNA expression of OPN in the two groups exposed to fluoride(1.89±0.37,1.94±0.22)was significantly higher than those of the corresponding control group(1.32±0.26,all P<0.05)and low-calcium group(1.30±0.186,P<0.05),respectively.High fluoride influenced the expression of protein and mRNA of OPN(F=13.821,4.24,all P<0.05).Low calcium did not affect the expression of protein and mRNA of OPN(F=2.164,0.58,all P>0.05).However,high fluoride and low calcium had a cross interaction on the expression of protein and mRNA of OPN(F=6.257,432,all P<0.05).Conclusions Over-dose fluoride enhances the expression of OPN.The higher expression observed in the cases exposed to high fluoride concentration is associated with serious renal injury.OPN may he a potential marker for renal injury in fluorosis.Moreover,over-dose fluoride and low calcium make the renal injury worse,indicating low calcium plays an important part in renal injury by fluoride.

Objective To investigate the changes of interleukin(IL)-2 and interferon(IFN)-γin serum and synovial fluid of patients with joint injure with Kashin-Beck disease(KBD), and study the roles of IL-2 and IFN-γ in KBD joint injure. Methods In accordance with the "Diagnostic Criteria of Kashin-Beck disease"(GB16003-1995),48 cases of KBD patients and 26 healthy people(control group) from KBD endemic area in Long county Shaanxi province were enrolled in the study. KBD patient were 24 males and 24 females, respectively, aged 40 to 65 years (mean age 51 years). Forty-eight serum specimens and 28 synovial fluid specimens of patients(14 males and 14 females,respectively) were collected. Healthy control group were 13 males and 13 females, respectively. Twenty-six serum specimens of healthy controls were collected. Serum and synovial fluid IL-2 and IFN-γ levels were determined by enzyme-linked immunosorbent assay(ELISA). Results In healthy controls and KBD patients, the midian of serum IL-2 were 46.8 ng/L and 55.7 ng/L, respectively, and IFN-γ were 52.3 ng/L and 48.8 ng/L, respectively. The difference was not statistically significant between healthy controls and KBD patients(t = 0.62, 0.70, all P ＞ 0.05).In synovial fluid of KBD patient, the midian of IL-2 and IFN-γwere 48.3 ng/L and 44.1 ng/L, respectively. The difference was not statistically significant between serum and synovial fluid in KBD patients(t = 0.69, 1.72, all P ＞0.05). Conclusion Serum and synovial fluid IL-2 and IFN-γare not significantly increased in KBD patients with articular damage, indicating that IL-2 and IFN-γare not involved in KBD joint injury.

BACKGROUND:Epidural scar after laminectomy is one important reason for the secondary spinal stenosis, and local application ofSanqi sodium hyaluronate gel can prevent epidural scar adhesion after laminectomy. OBJECTIVE: To study the effects ofSanqisodium hyaluronate gel on α-smooth muscle actin expression in the process of rabbit's epidural scar formation. METHODS: In this study, there were ninety-six rabbits which were randomized into four groups and given 0.5 mL normal saline, 0.5 mLSanqi concentrated solution, 0.5 mL sodium hyaluronate and 0.5 mLSanqisodium hyaluronate gel around the dura. At 1, 2, 4, 8 weeks after treatment, immunohistochemistry staining was employed for analysis of α-smooth muscle actin expression. RESULTS AND CONCLUSION:At the end of weeks 1 and 2, the expression of α-smooth muscle actin antibody in the normal saline group was significantly higher than that in the other three groups (P < 0.01 orP < 0.05), but there were no significant differences among the Sanqi, sodium hyaluronate andSanqisodium hyaluronate gel groups (P> 0.05). At weeks 4 and 8, the expression of α-smooth muscle actin antibody in theSanqi sodium hyaluronate gel group was significantly lower than that in the other three groups (P < 0.01 orP < 0.05), and there was no significant difference among the latter three groups (P > 0.05). These findings suggest thatSanqi sodium hyaluronate gel can inhibit the expression of α-smooth muscle actin, and thus ease scar contracture.

Human Interleukin-11 (hIL-11) is a multifunctional cytokine which plays an important role in regulating the proliferation and differentiation of cells in the hematopoeitic,lymphoid system etc. To obtain the IL-1 1 cDNA, a primary culture of Chinese fetal lung fibroblast was prepared from fresh tissue. Then the human IL-11 cDNA without the N-terminal signal peptide sequence was cloned by RT-PCR from the cells induced by PMA. The sequnce indicated that there are three bases different from those previously reported,but with no change of the amino acids. The cDNA was inserted into the 3′ end of trxcA gene in thioredoxin gene fusion expression system pTRXFUS to construct the trxA and hIL-1 1 fusion expression vector, and expressed in E. coli. The fusion hIL-11 accounts for more than 20 % of the total bacteria proteins.The expression product is present in soluble forms and has the full biological and immunological activities.

To analyze the structural characteristics of the Rap2B gene from Clonorchis sinensis by bioinformatics and to clone and express this gene through genetic engineering methods in order to obtain the corresponding recombinant protein.The structural and functional characteristics of the Rap2B protein were analyzed by using the related bioinformatic softwares. Using the full-length cDNA plasmid clone (Noc009e03) as template , the coding region of Rap2B gene sequence was amplified with PCR and cloned into the prokaryotic expression vector pET-28a(+) and then expressed in E.coli BL21 through IPIG induction. The recombinant protein expressed was purified by Ni-IDA affinity chromatography and detected by SDS-PAGE and Western blotting. It was demonstrated that the size of this gene was 847 bp in length, encoding 141 amino acids and its theoretical molecular weight was 15851.1 daltons. As demonstrated by PCR, double enzyme digestion and DNA sequencing, the recombinant expression plasmid pET-28a(+)-Rap2B was successively constructed and the Rap2B-like gene was highly expressed in E.coli BL21. The recombinant protein was obtained through purification with His affinity chromatography and could react specifically with the serum of rats infected with C.sinensis. Through these investigations, the Rap2B protein may be predicted as a member of the Ras oncogene family protein and is potential in the carcinogenic process of C.sinensis.

Objective To investigate the meaning of PURA gene and its protein in nephridial tissue of the rats with endemic fluorosis of coal burning. Methods Thirty-six SD rats of 80 - 100 g, body weight were randomly divided into control group, low fluorosis group and high fluorosis group according to body weight, 12 in each group, the number of female and male in each group was the same respectively. The control group, Low fluorosis group and high fluorosis group rots were fed with 1.5,25.0,60.0 mg/kg fluoride content in feedstuff, to establish the animal model of fluorosis. Expressions of both mRNA and its protein of PURA gene in rat nephridium tissue, were determined by RT-PCR and immunohistochemistry after four-month experimental period. Results The expressions of PURA mRNA[(2.74± 1.06),(4.29 ± 2.11)] and its protein[ (28 827.91 ± 4801.94),(61 146.96 ± 4997.55)] in low fluorosis group and high fluorosis group was higher than that in the control group[ ( 1.13 ± 0.87), (7131.95 ± 1524.54), all P < 0.05]. And the expressions of PURA mRNA and protein in high fluorosis groups was higher than that in low fluorosis greup(all P < 0.05). Conclusion High fluoride can lead to the high expression of PURA gene mRNA and protein in the rat nephridium tissue exposed to sodium fluoride.

The method of quantitative nuclear magnetic resonance spectroscopy (qNMR) for determination of epigoitrin in Radix Isatidis was established based on solid phase extraction (SPE).The twice ultrasonic extraction method using pure water was used for fully extracting epigoitrin in sample, and then the extraction was enriched and concentrated by poly-Sery MCX SPE cartridge.The effect of sample pretreatment and qNMR experimental conditions was investigated.The qNMR experiment conditions were selected using DMSO as solvent, calibrated 2,3,5-triiodobenzoate as internal standard, and P1(pulse width)=14.1 μs, d1(pulse delay time)=5 s, NS(number of scan)=256.The .1H-NMR peaks of δ 5.365-5.399 (H-7b, d, 1H) of epigoitrin were chosen as the quantitative peaks.Method validation was performed including precision (intra-day precision RSD was 0.5%, and the inter-day precision was 0.8%), linearity (correlation coefficient r>0.9991), LOD (0.05 mg/g, standard curve method) and LOQ (0.19 mg/g, S/N≥150).The recoveries of the SPE-qNMR were 97.4%-101.7%.The result showed that the method was stable, accurate and reliable.With this method the epigoitrin in a real Radix Isatidis was determined to be <0.19-1.26 mg/g.SPE combining with qNMR could extend the application field of qNMR, especially in the detection of low-content component in complex samples.

Objective To investigate the factors influencing basic rehabilitation service needs of people with disability at Zhabei District in Shanghai .Methods Primary rehabilitation demands and services of 10 311 disabled persons in Zhabei District were surveyed.Results The screening rate of persons with disability having primary rehabilitation demands was 83.5%. And the gender, age, type of disability, disability level, education, employment might be the factors affecting the primary rehabilitation demands of people with disability.Conclusion The primary rehabilitation service demands vary in deferent types of people with disability. The various services based on the extensive rehabilitation services should be provided to deferent categories of people with disability.

Objective To understand the status of special rehabilitation service demands of people with disability at Zhabei District in Shanghai and to provide the basis for the policy development.Methods Screen rate (SR) was used to indicate the level of demand of rehabilitation service, the data were analyzed with SPSS 13.0 statistical software.Results There were 4 349 persons with disability who demanded the special rehabilitation service; the SR of “on-site rehabilitation service” demands was the highest among the whole persons with disability; the SR of “personalized allotment of supplies and appliances” demands of the intellectual disability and mental disability was lower than the other types; the SR of special rehabilitation service demands of the female, the elderly, the low cultural level, the unemployed and the people with severe disability were relatively higher.Conclusion The form of “on-site rehabilitation services” needs to improve based on the demand of the disabilities; “personalized supplies and appliances” for the intellectual disability and mental disability needs to further develop; the rehabilitation services targeted to the female, the elderly, the low cultural level, the unemployed and the people with severe disability need to improve.

Objective To provide the advice on how to optimize the allocation of resources and make other policy through the analysis factors affecting rehabilitation services needs of people with disability at Zhabei district in Shanghai.Methods The factors affecting rehabilitation services needs such as types of disability, disability levels, education, employment, marital status, age and sex were analyzed with Logistic analysis.Results The factors affecting primary rehabilitation services were types of disability, education, employment situation, marital status and sex. And people with disability had the characters of female, hearing disability, retired returned to more primary rehabilitation services, with OR values being 1.219, 1.544 and 1.533 respectively compared with itself controls. And the high educational disable people' will, returning to primary rehabilitation services, was weaker compared with low educational disable people. The factors affecting special rehabilitation services were types of disability and employment situation. And people with disability had the characters of hearing disability, unemployment returned to more special rehabilitation services, with OR values being 2.380 and 1.310 respectively compared with itself controls.Conclusion The rehabilitation needs are different among different groups people with disability, so the factors affecting rehabilitation services needs such as types of disability, education, employment situation, marital status and sex should be taken into consideration while activating primary rehabilitation services, while the types of disability and employment situation should be taken into more consideration while activating special rehabilitation services.