Objective:To transfect human osteoprotegerin( o pg) gene into Cos-7 cells.Methods:The primers were designed b ased on the human opg cDNA sequence,total mRNA was isolated from 293 cells and RT-PCR was performed.The fragment of opg cDNA was inserted into pSecTag 2/B vector and sequenced by automatic sequence analyzer. The recombined plasmid was transfected into Cos-7 cells by Lipofectamine 2000 and OPG protein expressi on in Cos-7 cells was determined by immunohistochemistry and Western blot. Results:The sequence of opg cDNA from 293 cells obtained by RT- PCR was identical to the sequence provided by GenBank [gi:33878056]. The fragm ents of the recombinant plasmid digested with Hind Ⅲ,EcoR I and BamH Ⅰ and exa mined by 10 g/L agarose electrophoresis were consistent with predicted size.OPG over-expressing Cos-7 cells was selected and confirmed by immunohistochemistry and Western blot.Conclusion:Human opg gene may be transfect ed into Cos-7 cells.

Objective To study the variation of endothelin 1(ET 1) and calcitonin generelated peptide(cGRP) and its clinical implication, and to analyze the influence of ET 1 and cGRP on cerebral arteries hemodynamics in neonates with hypoxic ischemic encephalopathy(HIE). Methods The variation of ET 1,cGRP in cord blood and peripheral blood were prospectively observed at the time of 1 day,3 days and 7 days after birth in HIE and healthy control infants by radioimmuneassay. The cerebral arteries hemodynamics were determined immediately using color Doppler ultrasonography at 1 day after blood samples were obtained. Results (1) The following factors in cord blood and vein one day after birth in neonatal HIE and healthy controls was: ET 1(70?25)ng/L vs (38?19) ng/L ( P

The comorbidity of pain and depression is common. Both disorders might share common neuroanatomical and molecu-lar mechanisms. Recent studies have found that the brain-de-rived neurotrophic factor( BDNF) ,which plays an important role in the process of pain-depression comorbidity, has gradually be-come a hot topic and target of treatment. The article mainly sum-marizes the mechanism underlying comorbid pain and depres-sion,as well as the significance of blood BDNF in diagnosis and treatment of pain and depression.

Objective To explore the influence and its efficacy of Tong-Xinluo capsule on plasma endothelin levels in the patients with acute cerebral infarction(ACI).Methods 60 patients with ACI enrolled within 72 h of onset were allocated to Tong-Xinluo group(n=30) or control group(n=30) randomly.The control group was treated with Piracetam and aspirin,while Tong-Xinluo was provided 4 tablets tid for 30 d together with Piracetam and aspirin in the other group.Before and at 30th day after treatment,the patients were evaluated by European stroke scale(ESS) and activities of daily living(ADL),and the changes of ESS and ADL were served as the index of efficacy.The plasma level of endothelin-1(ET-1) was tested in the meanwhile.Results 30th day after treatment,there was a significant derease of ET-1 both in Tong-Xinluo group [(40.3?20.5)pg/ml] and control group [(31.5?14.7)pg/ml](P

Objective To observe the protective effect of Chinese traditional medicine Ginkgo Biloba extract on rat brain with cerebral ischemia/reperfusion.Methods Compared with ischemia/reperfusion group, Ginkgo Biloba extract-treated group was observed the change of ischemia and edema volume and NAA and lactate in by DWI and ~1H-MRS examination after cerebral ischemia /reperfusion 1 h,3 h,6 h. Results A significant reductions in infarct volume were found in the Ginkgo Biloba extract-treated group compared with the control group after cerebral ischemia 60 min/reperfusion 1 h,3 h,6 h. Ginkgo Biloba extract could decrease the concentration of Lac/(PCr+Cr) and prevent the decline of the concentration of NAA /(PCr+Cr) after cerebral ischemia/reperfusion, it had significantly difference compared with the control group(all P

Objective: To explore the signal transduction pathway of calcium-sensing receptor (CaSR) mediating hypoxia-induced proliferation of A549 cells of human non-small cell lung cancer. Methods: The A549 cells were randomly divided into several groups which conclude control group, hypoxia group (H), hypoxia + CaSR agonist group (H + Gd), hypoxia + CaSR inhibitor group (H + NPS) and phospholipase C(PLC) pathway inhibitor group (H + Gd +U73122). Expression of CaSR and proliferating cell nuclear antigen (PCNA) in A549 cells under different treatments was analyzed by Western blotting. The changes of intracellular calcium ion concentration were detected by confocal laser scanning microscope. The effects of cell proliferation cycle and proliferation index were gauged by flow cytometry under different drugs. HE staining was used to observe the changes of cell number with different drugs. Results: The expression levels of CaSR and PCNA in A549 cells increased by hypoxia. Meanwhile, cell proliferation index and cell number were also upregulated. GdCl3(CaSR agonist) could amplify the effect of hypoxia, and NPS2390 (CaSR inhibitor) could reduce the effect of hypoxia. All effects mentioned above could be inhibited by U73122 (PLC pathway inhibitor). Conclusion: Hypoxia-induced CaSR can mediate the proliferation of A549 cells through Gq-PLC-IP, signal transduction pathway.

Objectivo To analyze urine organic acids in the urine using gas chromatography-mass spectrometry(GC/MS)for diagnosis of inherited metabolic diseases,especially for organic acids metabolic disorders.Methods 195 clinical urine samples from the patients with suspected organic acids metabolic disorders and 5 normal urine from adults were collected.After mixing some urine with intemal standards according to the concentration creatinine and adding hydroxylamine hydrochloride to mixture,the organic acids with hydroxyl group were oximated to the ketobodies.Organic acids were extracted twice with ethyl acetate and ethyl ether and derivatized with BSTFA-TMCS.An the organic acids were determined with Agilent GC/MS 6890/5973i with scan model.the mass-to-charge ratio range is 50-550 m/z,all data were nalyzed with Agilent GCMSD ChemStationG1701DA.We also investigated the linearity, accurate,precision.recovery and Carry-over by determining the internal standards in normal samples and positive organic acids in spiked control samples.Results More than one hundred kinds of organic acids in urine samples can be analyzed with this method.According to the two internal standards in normal urine samples,the minimal detection limit MMA and 2.PA was 2.5-2.8 μmol/L.Intra-and interassay coefficient of variation for MMA and 2-PA are both less than 10%.Pre-processing Interassay coefficient by sequential preparations of the same sample was 14%.The recoveries of the spiked samples were 95%-105%.Carryover analysis was less than 1%.All the parameters meet the requirement for clinical diagnosis.12 samples demonstrated positive including 6 cases of methylmalonic acidemia,1 case of propionic acidemia,3 cases of tyrosinemia Ⅰ,1 case of maple syrup urine disease and 1 cases of ketosis.Conclusions The method for the determination of organic acids in urine by GC/MS has been successfully established.It can be used for clinical screening and diagnosis for inherited genetic metabolic diseases.

Objective To investigate the correlation between drug resistance and β-actamase genes of multi-drug resistance Acinetobacter baumannii (MDR-AB) in neonatal intensive care unit to provide evidence for rational antibiotics administration and nosocomial infection control.Methods Twenty-six MDR-AB strains were separated and collected from clinical specimens.The minimum inhibitory concentrations of 13 antimicrobial agents were determined by agar dilution method.Genotypes of β-lactamase were detected by polymerase chain reaction.Results The resistant rates of the 26 strains to Ceftazidime,Cefoxitin,Piperacillin-tazobactam and Ciprofloxacin were 100.0%.About 80.8% to 96.2% of these strains were resistant to the other antimicrobial drugs.Among the 26 MDR-AB strains,100% (26/26) strains possessed oxa-51,77% (20/26) possessed oxa-23 gene,54% (14/26) carried arnpC gene,both oxa-23 and ampC were identified in 42% (11/26) strains,while oxa-24,oxa-58,imp-1,imp-4 and vim-2 gene were not identified.Conclusions The drug resistance of Acinetobacter baumannii is serious,oxa-23 and ampC are the major plactamase genes carried by MDR-AB in neonatal intensive care unit.

Objective To carry out a meta-analysis on the published data in order to evaluate the clinical efficacy of prophylactic antibiotics in severe acute pancreatitis (SAP). Methods We searched the MEDLINE, EMBASE, Cochrane data base for systematic reviews and China biological medicine for randomized controlled trials on the efficacy of prophylactic antibiotics in patients with SAP from 1975 to May 2010. Eight studies met the inclusion criteria. Two authors independently extracted the data from these studies. The data were analyzed using the RevMan 4. 2. 10 software. Result In patients with SAP, prophylactic antibiotics did not reduce pancreatic infection (OR = 0. 67,95% CI:0. 43～1. 02,P = 0. 06), surgical intervention (OR = 0. 90,95% CI: 0. 60～ 1. 36 ,P = 0. 63) and mortality rate (OR = 0. 69,95% CI: 0. 41～1. 15,P=0. 16). Conclusion Antibiotic prophylaxis of SAP did not reduce mortality. It did not protect against pancreatic infection and the frequency of surgical intervention.

Objective The report was focused on polycystic ovary syndrome(PCOS) in female adolescents related to homosexual precocious puberty,and the new awareness of its influence on long-term health of homosexual precocious girls.Methods A girl with idiopathic central precocious puberty (ICPP) but diagnosed as PCOS in adolescence was reported and the data were reviewed.Results A girl was diagnosed as ICPP when she was near 8 years old with obesity,but without hyperinsulinemia,then she received the treatment of a 3.75 mg dose gonadotropin hormone analogues (GnRHa) every 28 days.Her gonads development was under control while her growth was arrested.Growth hormone(GH) injection started at 13 months.She stopped all medications when finished GH and GnRHa treatment for 12 months and 25 months.She went to see doctors again because of no menarche after discontinuing medication for nearly 2 years.She appeared obese and acne,hirsutism,athanens negricans and purple purple striae on the skin.Hyperinsulinemia and hypertestosterone were demonstrated.Pelvic B ultrasonography showed polycystic ovary,and she was diagnosed as PCOS.She was ordered to lose weight and to take metformin.And adrenocorticotropic hormone stimulating test was done,and B ultrasonography again ruled out atypical congenital adrenal hyperplasia and tumor of adrenal gland.She got her menarche 1 month later.Twelve months after the PCOS diagnosis and treatment,she had 4 menses,her insulin level decreased,glucose tolerance and her serum testosterone level turned normal.At the same time,the form of ovarian and follicular was significantly reduced.After taking mefformin for 20 months,her height and weight did not change,her menstrual was regular every 35-40 days,each time lasting about 7 days.She was followed up.We also reviewed literatures and learnt that precocious puberty might not be a benign situation and it might have an intrinsic relation to obesity,precocious puberty and PCOS.It might be the cofactors for causing PP and PCOS that accessed luteinizing hormone(LH) secretion and disorder of hypothalamus pituitary gonad(HPG) axis except obesity with hyperinsulinemia.Reported pubertas praecox in childhood developed to PCOS at 30％ with high prevalence of obesity.GnRHa suppressive therapy might relate to PCOS and had disputation for improving final height.There were fewer investigations on adolescent PCOS,and no consensus guideline on it in China.Conclusions Girls with ICPP may develop to PCOS some time later.The clinical features of the reported girl and the knowledge from literatures support the hypothesis that inner relationship between the CPP and the PCOS.The LH high secretion and disorder of HPG axis may be the causes of them; LH treatment on suppressing obesity should be cautious during treatment.The benefit and risk from GnRHa treatment should be evaluated thoroughly.Further clinical research should be conducted on adolescent PCOS.