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 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.

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 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

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 analyze the clinical features of 35 patients with Silver-Russell syndrome (SRS) in Beijing Children's Hospital in recent 6 years,and to recognize the clinical manifestations and improve the diagnosis of the disease.Methods The diagnostic criteria of SRS were studied retrospectively,the differences among 3 different diagnostic criteria were compared.Results Thirty-five SRS patients were diagnosed by means of 3 criteria,over a period from 2006 to 2012.There were 23 males and 12 females,with age range 0.08 ～ 12.15 yr.77 ％ of cases met the three criteria,97％ met two of the three criteria.There were dwarfism in 33 cases (94.2％),genital abnormalities in 1 case (2.9％),and asymmetry of limbs in 1 case (2.9％).The clinical characteristics with the frequencies accounting for over 80％ included short stature (average height-3.61 SDS),leanness (body mass index ＜-2.11SDS),bone dysmorphism including triangular face,irregular teeth,asymmetry of limbs,clinodactyly of fifth finger,and bone age obviously lagged behind.60％ of the cases showed bone age lagging behind more than 1 year.Height lagging behind was more obviously then bone age.Conclusions The diagnosis of SRS is mainly based on Clinical manifestation,laboratory and biochemical examinations are of no avail.There is a high coincidence with the 3 different diagnostic criteria.Clinical features in more than 80％ patients with SRS are short stature,craniofacial triangular dysmorphism,irregular teeth,limbs asymmetry and fifth finger clinodactyly,and mild lagging behind of bone age.

Objective To explore the blood drug concentration monitoring of sustained-release valproate(DK)in children with epilepsy,focusing on the selection of sampling time and evaluation of the results.Methods Two hundred and seventy-one children taking DK and 155 children taking sodium valproate syrup(VPA Syr)were involved and their serum were taken when achieved steady state to determine the valproic acid level using fluorescence polarization immunoassay.They were divided into 4 groups,which were DK taken once daily group(DK qd group,126 children),DK taken once daily at night and sampled on morning group(DK qn group,26 children),DK taken every 12 h group(DK q12 h group,119 children),VPA Syr q12 h group(155 children).Determine the proportion of the blood drug concentration of each group below,ithin and above the therapeutic range for valproate(50-100 mg/L)were determined.The data were analyzed by t test.Results The Cmin of DK qd group were(73.09?19.91)mg/L,significantly lower from the serum concentration of DK qn and sampled on morning group [(94.94?25.44)mg/L](P0.05).Conclusions DK qn should sampled at night before the night dose.The Cmin of DK q12 h was higher according to the therapeutic range,it's favorable range still needs clinical practice.

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.

Aim To observe the neuroprotective effect of platelet activating factor receptor antagonist Kadsurenone on rat brain with cerebral ischemia/reperfusion.Methods Animal model was established through middle cerebral artery occlusion(MCAO).we studied the changes of infarct volume,NAA and Lactate in Kadsurenone-treated group with DWI and()~1H-MRS examination after cerebral ischemia 60 min and reperfusion 1,3,6 h.Results Significant reductions in infarct volume were found in the Kadsurenone-treated group as well as in the Ginkgolides-treated group compared with the control group.And Kadsurenone decreased the concentration of lactate and prevented the decline of the concentration of NAA after cerebral ischemia/reperfusion.Conclusion As platelet-activating factor receptor antagonists,both Kadsurenone and Ginkgolides show remarkable neuroprotective effect.And MRI offers exact neuroimaging information for studying the neuroprotective meachnism of Chinese traditional medicine after cerebral ischemia and reperfusion.