Objective:To obtain human S100A6-GST fusion protein as material of the study of function of hS100A6.Methods:The hS100A6 gene from pHAHA-hS100A6 was subcloned into the prokaryotic GST fusion protein expression plasmid,pGST-moluc,to form pGST-moluc-hS100A6. The recombinant plasmid was transformed to E.coli BL2l and the expression of hS100A6-GST fusion protein was induced with IPTG. The protein was detected by SDS-PAGE and purified with Glutathion-Sepharose 4B beads,then identified by western blot assays. Results:A 36 000 Dalton protein,as expected,was obtained evidently.The concentration of the purified fusion protein was 3mg/L bacterial culture,and the purity was about 92%. Conclusion:A prokaryotic system expressing hS100A6-GST fusion protein was successfully constructed.hS100A6-GST fusion protein with high purity could be obtained after it was efficiently expressed in E.coli BL21 and purified with Glutathion-Sepharose 4B beads. This will facilitate our study of the function of hS100A6.

Aim To investigate the antiepileptic activity of alpha-asarone in three epilepsy models.Methods The MES mice,MST mice and Lithium-pilocarpine rats were divided randomly and respectively into groups each containing 20 animals(?-asarone groups,AED groups and normal control group).Different doses of alpha-asarone were administered to mice/rats in advance in alpha-asarone treated groups,one group received only saline,while the other groups received antiepileptic drug as a reference standard,2 times per day for 28 days.The seizure severity score,seizure latency and total number of animals with seizures were noted to observe whether alpha-asarone had anticonvulsant effect or not in three epilepsy models.Results Alpha-asarone possessed excellent anticonvulsant effect in MES and MST and lithium-pilocarpine models. It significantly decreased the seizure incidence 40%~100% in the MES models and 50%~90% in MST models,and 40%~80% in the Lithium-pilocarpine model. It significantly prolonged the seizure latency 70~180 s in MST mice and 4~15 min in Lithium-pilocarpine rats;It significantly reduced the seizure severity scores 1.96 in Lithium-pilocarpine rats.Conclusions Alpha-asarone had a positive antiepileptic activity.

Objective To dynamically observe the effect of compound decotion on changes of Na+-K+-ATPase,Ca2+-ATPase activity,intracellular free Ca2+ contents and CaM expression jn bomogenate and mitochondria of rat brain tissues after traumatic brain injury(TBI)and investigate the molecular mechanism of neuroprotective effect of compound decotion.Methods Rat TBI models were made and divided into sham operation group,TBI group and compound decotion treatment group(treated with comof normal saline,twice per day for seven days.Rats were sacrificed at 24 h,72 h and 1 week after injury to dynamically observe activities of Na+-K+-ATPase and Ca2+-ATPase in bomogenate and mitochondria of rat brain tissues,concentration of free Ca2+ in neurocytes and expression change of CaM in brain tissues.Results The activities of Na+-K+-ATP ageand Ca2+-ATPase in homogenates and mitochondria of brain tissues markedly decreased at different time point and increased gradually after 72 hours in TBI group but wag still lower than that of sham operation group at one week after injury.However,compound decotion could significantly enhance the activities of Na+-K+-ATPageand Ca2+-ATPage(P＜0.05).In TBI group,concentration of free Ca2+ in neurocytes and CaM expression in brain tissues were elevated at different degrees at different time point and reached peak at 24 hours after injury but still lower than that of sham operation group at 72 hours.While concentration of free Ca2+ in neurocytes and CaM expression in brain tissues were significantly lower than those of TBI group at different time point(P＜0.05).Conclusions The neuroprotective effect of compound decotion may be related to its role in increasing activities of Na+-K+-ATPase and Ca2+-ATPase to facilitate cellular metabolism and decreasing concentration of free Ca2+ in neurocytes and CaM expression in brain tissues to mitigate secondary brain injury induced by Ca2+ over load.

Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is an autosomal recessive disease caused by SLC25A13 gene mutations, and is characterized by delayed jaundice clearance, liver dysfunction, and elevated aminoacidemia. The confirmed diagnosis depends on gene analysis. Citrin deficiency is one of the important causes of neonatal intrahepatic cholestasis in China. Recently more and more researches about NICCD were reported. The paper summarized the epidemiology, pathogenesis, clinical characteristics, and progresses in diagnosis and treatment of NICCD.

Objective The biological effects of BMP3 on osteosarcoma were investigated by treating human osteosarcoma cell lines MG63,and U2OS with human BMP3(hBMP3).Methods Osteosarcoma cells in experimental groups were respectively treated with AdBMP-3 and rhBMP3-CM,control groups with AdGFP and rGFP-CM,the blank group with neither.Their ability of proliferation,apoptosis,transmigration and differentiation were respectively detected by trypan blue exclusion test,terminal deoynucleotidyl transferase mediated dUTP nick end labeling(TUNEL) and acridine orange-ethidium bromide fluorescent stain(AO/EB),transwell-room and alkaline phosphatase(ALP) activity reagent kit.Results(1) All the indexes detected were not significantly different between two control groups.(2) Compared with control groups,the cell survival rate showed a significant decrease in experimental groups.(3) The apoptosis indexes increased.(4)The number of trans-membrane cell decreased.(5)The activity of alkaline phosphatase increased after treatment with AdBMP3 and rhBMP-3 for 3 days in MG63,5 days in U2OS.Conclusion hBMP3 inhibit osteosarcoma cells growth and promote bone formation.

OBJECTIVE: To investigate the effect of human bone morphogenetic protein (hBMPs) 2/3/6 and 12 on osteosarcoma cell UMR106. METHODS: Adenovirus-BMP2/3/6 and 12 (AdBMP2/3/6 and12) were used to treat the cell line. Their proliferation, apoptosis, and transmigration were detected by Trypan blue exclusion test, TdT-mediated biotinylated-dUTP nick end labeling (TUNEL), acridine orange-ethidium bromide (AO/EB) double fluorescent dye staining, and transwell-room test, respectively. The alkaline phosphatase (ALP) activity was detected to reflect the differentiation of tumors. RESULTS: Compared with the control groups, the cell survival rate of the experimental groups treated with AdBMP2/3/6 and 12 showed a significant time-dependent decrease (P<0.01). The apoptosis indexes were increased significantly (P<0.01) and the results from TUNEL and AO/EB method were consistent. The cell numbers of transmembrane significantly decreased at 24,48, and 72 h (P<0.01). AdBMP2/3/6 and 12 treatment enhanced the activity of ALP activity from day 3 and this effect might still be observed up to day 9 of the treatment (P<0.01). CONCLUSION hBMPs2/3/6 and 12 can inhibit the proliferation and transmigration, and induce their apoptosis and differentiation in osteosarcoma cell line UMR106.
Adenoviridae ; genetics ; metabolism ; Apoptosis ; drug effects ; Bone Morphogenetic Protein 2 ; pharmacology ; Bone Morphogenetic Protein 3 ; pharmacology ; Bone Morphogenetic Protein 6 ; pharmacology ; Bone Morphogenetic Proteins ; pharmacology ; Bone Neoplasms ; pathology ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; drug effects ; Growth Differentiation Factors ; pharmacology ; Humans ; Osteosarcoma ; pathology ; Recombinant Proteins ; pharmacology

Objective To explore and analyze the clinical characteristics of extremely premature infants at low risk for early-onset sepsis (EOS) , so as to avoid overuse of antibiotics. Method The clinical data of extremely premature infants hospitalized from January 1, 2010 to December 31, 2017 were collected. Extremely premature infants born from mothers without premature rupture of membranes and without maternal clinical manifestations of chorioamnionitis during pregnancy were classified assigned into the low-risk group, and those who did not meet the low-risk conditions were regarded assigned intoas the control group. EOS was diagnosed according to the results of blood culture within 72 hours after birth. The clinical characteristics, treatment and outcome of extremely premature infants between the two groups were retrospectively analyzed. Results A total of 245 extremely preterm infants were enrolled, including 153 (62.4％) in low-risk group. Compared with the control group, mothers in low-risk group had higher rates of gestational diabetes and hypertension, higher rates of antenatal hormone use and lower rates of antenatal antibiotics use; furthermore, neonates in low-risk group had lower rates of Apgar score ＜ 5, higher rates of pulmonary surfactant use, respiratory support and mechanical ventilation, and lower risk of death and incidence of early-onset sepsis. The differences were statistically significant (P＜0.05) . Among In extremely premature infants whose having survival time＞ 24 hours, compared with control group, infants in low-risk group had higher incidences of respiratory distress syndrome, patent ductus arteriosus, intracranial hemorrhage and bronchopulmonary dysplasia, and lower incidence of pulmonary hemorrhage than control group, and the differences were statistically significant (all P＜0.05) . In low-risk group, the risks of death, respiratory distress syndrome, pulmonary hemorrhage and bronchopulmonary hemorrhage in long-term antibiotic group were higher than the short-term antibiotic group. Conclusion Early identification of extremely preterm infants at low risk of early-onset sepsis in extremely preterm infants is of clinical significance in reducing early empirical use of antibiotics therapy.