BACKGROUND:Estrogen/progestins replacement therapy prevents excess bone loss in postmenopausal women.Recently osteoprotegerin (OPG) has been identified in osteoblast and displayed to inhibit bone resorption.OBJECTIVE: To compare the action between 17β-estradiol (E2) and progesterone on OPG expression in cultured normal human osteoblast-like cells (hOB).DESIGN: A comparative investigation.SETTING: Institute of Metabolic Endocrinology, the Second Xiangya Hospital of Central South University.MATERIALS: α-MEM (Sigma Chemical Corp., St. Louis, MO, USA); Type Ⅳ collagenase (Sigma); Fetal bovine serum (Gibco-BRL Corp., Grand Island, NY, USA); Osteocalcin radioimmunoassay kit (DiaSorin Corp., Stillwater, MN, USA).METHODS: The experiments were carried out in the Institute of Metabolic Endocrinology, Second Xiangya Hospital of Central South University from January 2003 to March 2006. The osteoblasts were extracted from the cancelous bone of anterior superior iliac spine of normal people, then cultured. The hOB were treated with E2 and progesterone, and the expressions of OPG mRNA and OPG protein were determined by Northern blot analysis and enzyme-linked immunoabsorbent assay (ELISA) respectively.MAIN OUTCOME MEASURES: ①Characterization of human osteoblast-like cells; ②Effect of E2 and progesterone on OPG mRNA levels by Northern blot analysis; ③ Effect of E2 and progesterone on OPG protein levels in the conditioned medium by ELISA.RESULTS: ① Characterization of hOB in vitro The ALP levels in normal human osteoblasts were (74.3±4.7) U/g protein,and the detectable osteocalcin levels was (3.84±0.39) μg/L protein], which suggested that osteoblasts were the primary cell type found in our bone-derived cell cultures from donors. ② Effects of E2 and progesterone on the levels of OPG mRNA by Northern blot analysis: The OPG mRNA band was week in the control group [(12.3±3.5)%], treatment with 1 × 10-10, 1 ×10-9 1 ×10-8 mol/L E2 caused an increase in the levels of OPG mRNA. The expression of OPG mRNA in the 1×10-8 mol/L E2 group was gradually increased at 12, 24 and 48 hours. Progesterone had no influence on OPG mRNA expression. ③ Effects of E2 and progesterone on OPG protein production in conditioned medium determined with ELISA:ELISA revealed that treatment with 1 ×10-10, 1 ×10-9, 1 ×10-8 mol/L E2 induced obvious increase in the levels of OPG protein in cells media as compared with that in the control group [(1.27±0.26), (2.34±0.35), (3.62±0.23), (0.64±0.14)μg/L, P ＜ 0.01]. In the presence of 1×10-8 mol/L E2, OPG protein production in cells media at 12, 24 and 48 hours were significantly higher than that in the control group [(1.30±0.30), (3.07±0.14), (3.50±0.33), (0.62±0.12) μg/L, P ＜ 0.01]. 1 × 10-10, 1 ×10-9 1 × 10-8 mol/L progesterone had no influence on the OPG protein production after 12-24 hours (P ＞ 0.05).CONCLUSION: The different regulation of OPG production in osteoblasts by E2 and progesterone may contribute to the mechanisms by which estrogen or progestins exerts its different action on bone resorption.

Objective To investigate clinical effect and safety of ostium secundum atrial septal defect(ASD) treatment via one -stop hybrid and classical surgical procedures.Methods 45 patients were diagnosed ostium secun-dum simple ASD by ultrasound cardiogram and clinical manifestation,they were divided into one -stop hybrid proce-dure group (n =20)and classical surgical procedure (n =25).Age,gender,weight,post operation hospital day,on -pump time,blood transfusion amount,drainage flow,incision length and incidence of complication between the two groups were compared.Results Age and weight had no difference between the two groups(t =0.40 and 1.64,P >0.05),but the proportion of female cases in one -stop hybrid procedure group was higher than post operation(χ2 =9.45,P <0.05).Hospital day was shorter(t =11.11,P <0.05),drainage amount was fewer(t =81.68,P <0.05), incision length(t =22.51,P <0.05)was shorter.Incidence of complication had no statistically significant difference between the two groups(χ2 =0.35,P >0.05).And one -stop hybrid procedure group was off -pump without blood transfusion.Conclusion One -stop hybrid procedure was simple,could make a quick recovery post operation and was an ideal method for ostium secundum ASD treatment.

Cell separation technology is an important means for cell sorting and cell-population purification. It is the current international research hot spot in biochemical analysis which has very important applications in many fields such as biology, medicine, agriculture and environment. This review introduces the development status of cell separation using microfluidic chip based on dielectrophoresis. It expounds the working principle of dielectrophoresis and the key factors such as cell size, electrode shape and signals that impacts the dielectrophoresis of cell separation on different types of microfluidic chip. Finally, it forecasts the future development trend of cell separation using microfluidic chip based on dielectrophoresis.

18 fungal strains including N.coenophialum,N.lolii, N.huerfanum、N.chisosum、N.aotearoae、N.sp.and 8 varieties of grass seeds belonging to Festuca arundinacea and Lolium perenne have been studied.With amplification of IS1～IS3 and F1～R1 of genomic DNA, the primers Tub-2-F～Tub-2-R from Tubulin-2 gene and F3～R3 from NC25 gene have been designed.A PCR method to detect N.coenophialum and N.lolii was established, and also a nested-PCR method to detect N.coenophialum and N.lolii in single seed was established.These PCR detection methods are strongly special and much credible and rapid-speeded.

Immunohistochemical technique was an essential tool of conventional diagnosis,therefore,the application of immunohisto- chemistry in reformation of pathology laboratory teaching would boost pathological experimental teaching standards to a higher level.

Objective To investigate the iodine nutritional status of targeted population in the high-risk areas of iodine deficiency disorders in Chongqing, so as to provide scientific evidence for establishing prevention and remedial measures. Methods Six towns were selected in Chengkou and Wuxi Counties to found suspected dementia patients born after first Jan, 1997. Two hundred children aged 8-10 years were investigated in every town. The thyroid volume, intelligence quotient(IQ) and urinary iodine of the children were examined. Forty women (pregnant and nursing women) were investigated in every town. The iodine content of salt from their home was measured. The thyroid volume was examined by palpation and B-uhrasound. IQ was measured by combined Raven Test in China(CRT-RC2). Urinary iodine was determined using the acid digest arsenic-cerium contacting method, and iodined salt was detected using direct titration method. Results Six suspected dementia patients were found in the local town hospital. Five eases were excluded. There was 1 case born in other place. The rates of goiter by palpation and B-ultrasound were 9.58%(92/960) and 8.89%(65/731), respectively. The median of urinary iodine of children and women was 319.15 μg/L and 248.42 μg/L, respectively. The mean of IQ of the children was 103.32. The coverage rate of iodine salt from residents was 98.82%(336/340). Conclusions The iodine nutrition of children was good and there is no newly occurred cretinism in Chengkou and Wuxi Counties. Goiter rate and median of urinary iodine aged 8-10 years and of women, coverage rate of iodine salt from resident has meet the standard set for basical elimination iodine deficiency disorders.

To establish a fast sensitive, reproducible LC-MS/MS method to study pharmacokinetic properties of THC, and compare relative bioavailability of THC and its solid dispersion in mice. 200 mice were divided randomly into two groups, and administered orally with THC and THC-solid dispersion after fasting (calculate on THC:400 mg x kg(-1)), used HPLC-MS/MS method to determine the THC concentration of each period at the following times: baseline ( predose ), 15, 30, 45 min, 1, 1.5, 2, 3, 4, 6, 24 h after dosing. Calculating the pharmacokinetic parameters according to the C-t curv, and then use the Phoenix WinNonlin software for data analysis. The calibration curves were linear over the range 9.06-972 microg x L(-1) for THC (R2 = 0.999). The limit of detection (LOD) was 0.7 microg x L(-1), respectively. The average extraction recoveries for THC was above 75%, The methodology recoveries were between 79% and 108%. The intra-day and inter-day RSD were less than 13%, the stability test showed that the plasma samples was stable under different conditions (RSD < 15%). The precision, accuracy, recovery and applicability were found to be adequate for pharmacokinetic studies. Pharmacokinetic parameters of THC and THC-solid dispersion orally to mice shows as fllows: T(max), were 60 and 15 min, AUC(0-t) were 44 500.43 and 57 497.81 mg x L(-1) x min, AUC(0-infinity) were 51 226.00 and 68 031.48 mg x L(-1) x min, MRT(0-infinity) were 596.915 6, 661.747 7 min, CL(z)/F were 0.007 809 and 0.005 88 L x min(-1) x kg(-1). Compared with THC, the MRT and t1/2 of the THC-solid dispersion were all slightly extended, the t(max) was significantly reduced, AUC(0-24 h), AUC(0-infinity) and C(max) were all significantly higher, the relative bioavailability of THC-solid dispersion is 1.34 times of THC. The results of the experiment shows that the precision, accuracy, recovery and applicability were found to be adequate for the pharmacokinetic studies. After oral administration to mice, the relative bioavailability of THC-solid dispersion show significant improvement compared to THC.
Administration, Oral ; Animals ; Biological Availability ; Dronabinol ; administration & dosage ; chemistry ; pharmacokinetics ; Female ; Male ; Mice

Purpose To compare the difference of the genotype and allele of endothelial cell protein C receptor (EPCR)gene rs9574 C/G between Guangxi population and other ethnic groups.Methods The rs9574 C/G polymorphisms of EPCR in 130 cases of Guangxi population were detected by PCR and DNA sequencing.The distribution frequency of allele and genotype was compared with the other four ethnic groups (HapMap-CEU,HapMap-HCB,HapMap-JPT,HapMap-YRI),which was published by the human genome project.Results Three genotypes of CC,CT and TT were found in rs9574 C/G with the frequencies of 39.2％,46.2％,14.6％ respectively.the allele frequencies of C,T were 62.3％ and 37.7％.No significant difference was observed in the frequency of genotype and allele between male and female (P ＞ 0.05).There were significant differences in the genotype distribution among Guangxi population,HapMap-CEU and HapMap-YRI (P ＜ 0.05).Significant differences of allelic frequency were found among Guangxi population,HapMap-CEU,HapMap-JPT and HapMap-YRI.Condusion The polymorphisms of rs9574C/G in 3'-noncoding region of EPCR gene in Guangxi population were different in different regions and ethnic groups.

Purpose　To describe clinicopathological and immunophenotypic features of 10 cases of histiocytic necrotizing lymphadenitis (HNL). Methods　HE sections of 11 lymph node biopsies were re-examined. Immunophenotyping and detection of apoptotic DNA fragments were performed using S-P and TUNEL methods, respectively. Results　Five cases have been diagnosed as non-Hodgkin lymphoma. Histologically variable-sized discrete or confluent nodules were seen in the paracortex, especially in the interfollicular area, which were composed of proliferative pleomorphic histiocytes, transformed lymphocytes, and karyorrhectic debris. Immunohistochemistry revealed CD3+ and CD45RO+ for lymphocytes, Mac387+ and/or CD68+ for histiocytes, and no expression for CD15,CD30 and CD20 in the lesions. Conclusions　The presence of pleomorphic histiocytes, transformed T-cells, and karyorrhectic debris in the biopsy of lymph nodes, together with the absence of neutrophils support the diagnosis of HNL.

Objective To study the analgesic and anti-inflammatory effects in vivo of the total alkaloids in Stephania kwangsiensis (TARS).Methods Mice were respectively ig with high,middle,and low doses of TARS.The analgesic effects of TARS were evaluated by the formaldehyde induced pain method and tail-immersion response induced by hot water in mice,and anti-inflammatory effects were observed by cotton ball granuloma,egg white induced paw swelling method in mice.Results TARS could significantly inhibit formaldehyde induced pain in mice,reduce the hind licking times,extend tail withdrawal time induced by hot water,obviously reduced the formation of granulomatous in mice;Inhibited egg white induced paw swelling.Conclusion TARS has significant analgesic and anti-inflammatory effect.