BACKGROUND:Studies have shown that genetical y modified Schwann cel s can survive for a longer time in vivo, and promote nerve regeneration and functional recovery. OBJECTIVE:To transfect human telomerase reverse transcriptase (hTERT) gene into rat Schwann cel s cultured in vitro via PLXSN vector, and to detect the telomerase activity and biological characteristics of Schwann cel s. METHODS:Schwann cel s from Wistar rats were cultured in vitro and transfected by PLXSN vector with (hTERT group) or without hTERT (empty vector group). Normal Schwann cel s were selected as control group. RT-PCR and western blot methods were used to detect the hTERT protein and mRNA levels in Schwann cel s, and flow cytometry was used to measure the cel cycle distribution. Cel growth was observed by cel growth curve and MTT colorimetric method. RESULTS AND CONCLUSION:At 48 hours after transfection, the mRNA and protein expressions of hTERT were remarkably seen in Schwann cel s. Compared with the control and empty vector groups, the cel s grew faster, the number of cel s at G 0/G 1 Schwann cel s cultured in vitro. phase was reduced, but the number of S phase cel s was increased in the hTERT group (P<0.05). These findings indicate that PLXSN vector-mediated hTERT transfection of Schwann cel s can significantly improve the activity of telomerase in Schwann cel s as wel as promote the proliferation of Schwann cells cultured in vitro.

Objective: To study the mechanism of effect of total flavone of rhizoma drynariae in groups with different dosage on osteoblasts cultured in vitro. Methods: To culture osteoblasts with UMA-106-01 osteoblast strain and to observe the activity of ALP and the endosmosis of 3H-TdR. Results: The total flavone of rhizoma drynariae increased the activity of ALP in cells cultured with UMR-106-01 strain. The ALP activities correspondingly changed in the 24h, 48h and 72h, which related to dosage effect and time effect. Among them, the activity in the 48h is the most ideal one. And the amount of endosmosis of the total flavone of rhizoma drynariae to 3H-TdR increased more remarkably in the 48h than that in the 24h and it also related to the time effect. Conclusion: The total flavone of rhizoma drynariae can promote the differentiation and multiplication of osteoblasts.

Objective We tried to investigate the current status of pediatric nursing post setting and analyze the practical application effect in the clinical working practice for the foundation of readjusting and setting up the pediatric nursing positions.Methods By convenient sampling method,the general condition and current post setting situation in the pediatric wards of 7 hospitals with self-designed questionnaires were investigated.The results underwent analysis Results The nursing post setting in pediatric wards of 7 hospitals were not all the same,nursing posts in the pediatric ward included three categories:management positions,teaching positions and clinical positions.Management positions included the head nurse,deputy head nurse and head nurse assistant; teaching positions included the teaching secretary and clinical total teaching nurse; the seven kinds of clinical nursing positions were office job,day shift,night shift,evening shift,midnight shift,late-morning shift,auxiliary job.Though some nursing jobs differed in the position names and working hours,their job contents were nearly identical.Conclusions Pediatric nursing post setting management in our country is still in the exploratory stage at present,it has not form a unified perfect job management system.Pediatric nursing positions should be set from the perspective of evidencebased theory,satisfying the demands of patients as the guidance,coming from clinical needs as the first principle,in the full implementation of the accountability for nursing and based on comprehensive consideration of various factors,so that the post setting can be scientific and reasonable.

Objective To analyze the characteristics and the related risk factors of the contractility of pelvic floor muscles in old women with pelvic floor dysfunction(PFD).Methods A cohort of 125 elderly patients with symptoms of pelvic floor dysfunction,including 60 of postpartum urinary incontinence(SUI)and 65 of pelvic organ prolapse(POP),were recruited.60 women volunteers with normal pelvic floor function were recruited as a control group.A questionnaire survey was conducted to record the general situation of the subjects.The pelvic floor muscle strength was determined by the methods of electromyography and pressure feedback recording.Results The body mass index,the average number of delivery,newborn baby's body weight were significantly higher in the SUI and POP group than in control group,with statistically significant difference(F=5.29,6.27,5.26,P=0.007,0.003,0.008),but without statistically significant difference between SUI and POP groups (P =0.674,0.554,0.578).The rapid contractility value was significant lower in SUI group than in POP and control groups(all P＜0.01),and the sustained contractility value was significant lower in POP group than in SUI and control groups(all P＜0.01).Conclusions The postpartum pelvic floor dysfunction may be correlated with BMI,vaginal delivery rate,the average number of delivery,newborn baby's body weight and a history of increased abdominal pressure.The postpartum pelvic floor dysfunction may be associated with the declined contractility of pelvic floor muscle,among which the POP may be associated with class Ⅰ muscle and the SUI with class Ⅱ muscle.

Objective To investigate the apoptosis induction effects and the possible mechanism of YM155 on triple negative breast cancer MDA-MB-231cells.Methods MDA-MB-231 cells were treated with dif-ferent concentrations of YM 155, and the survival rate of the cells was determined by CCK-8 assay and the half maximal inhibitory concentration ( IC50 ) value of YM155 was calculated .The apoptosis rate was examined by An-nexin V-FITC/PI double staining.mRNA expression of survivin and bcl-2 in MDA-MB-231cells was detected by RT-PCR.The protein expression of survivin , bcl-2, caspase-3, and PARP were detected by Western blot .Re-sults YM155 significantly inhibited the growth of MDA-MB-231 cells in a dose-and-time-dependenct way .IC50 was(1.749 ±0.265) ng/ml and(0.823 ±0.125) ng/ml respectively at 24 and 48 hours.The apoptosis rate of cells treated with 0.5 ng/ml, 1.0 ng/ml, and 1.5 ng/ml YM155 was (10.93 ±0.94)%,(31.10 ±1.51)%, and(46.83 ±2.92)%respectively, which had significant difference compared to that of the control group (6.4 ± 1.2)%(P<0.01).YM155 could significantly decrease mRNA and protein expression of surviving , besides, it reduced bcl-2 expression and increased caspase-3 and PARP protein expression .Conclusions YM155 can ef-fectively induce the apoptosis of MDA-MB-231 cells by downregulating survivin and activating caspase pathway . Bcl-2 might play a role in the apoptosis .

Objective:o prepare Danshensu liposomes and investigate drug release characteristics in vitro. Methods: Danshensu liposomes were prepared by a reverse-phase evaporation method. The encapsulation efficiency was used as the index, an orthogonal test was adopted to investigate the effect of concentration of soybean lecithin, ratio of lipid-Danshensu and pH value of solution on the preparation procedure of Danshensu liposomes. The particle size of the liposomes was also investigated by a transmission electron micro-scope ( TEM) . The concentration of Danshensu was determined by HPLC, and the difference of release characteristics in Danshensu li-posomes and Danshensu solution was measured by a dialysis method. Results:The optimum preparation technology was as follows:the concentration of soybean lecithin was 40 mg·ml-1 ,the ratio of drug-lipid was 1: 10,and the pH value of solution was 6. 6. The mor-phology of the prepared liposomes showed spheric structure with uniform diameter, and the average particle size was ( 174 ± 36 ) nm and the encapsulation efficiency was 38. 9%. The linear range of Danshensu was 2. 0-20. 0 mg·L-1(r=0. 9984). The drug release of liposomes in vitro was slower than that of free Danshensu solution in 24 h. Conclusion:Danshensu liposomes with fine morphology have sustained release property.

HBV polymerase has intrinsic RNA-dependent reverse transcriptase, DNA-dependent DNA polymerase as well as RNaseH activity. Analysis of HBV polymerase has been hampered for many years due to the inability to express functional enzyme in a recombinant system. To obtain active polymerase at a high level, we have taken advantage of baculovirus expression system. The gene of HBV polymerase was amplified by PCR and cloned into pFastBac Dual to construct the recombinant plasmid pFastbac Dual-pol. The recombinant donor plasmid, pFastbac Dual-pol, was constructed by inserting HBV polymerase gene into EcoRI and PstI sites controlled by polyhedrin promoter. The recombinant donor plasmid was transformed into DH10Bac competent cells for transposition. Recombinant bacmid was constructed by inserting of the mini-Tn7 element from the donor plasmid into the mini-attTn7 attachment site on the bacmid. The recombinant bacmid DNA was isolated and transfected into the Sf9 cells to produce the recombinant virus, and healthy insect Sf9 cells were infected with the recombinant virus containing HBV polymerase gene to express the target protein. HBV polymerase expressed in insect cells was analyzed by SDS-PAGE. PCR results showed recombinant donor plasmid, pFastbac Dual-pol, was constructed successfully. The recombinant hepatitis B virus polymerase was expressed in insect cells at high level. The recombinant hepatitis B virus polymerase should facilitate the analysis of HBV polymerase biological characteristics, allow the investigation for new anti-HBV drugs specifically blocking HBV polymerase.

The data of 10 cases of traumatic extra-articular ankylosis of temporomandibular joint(TMJ),including the type of trauma and the type of ankylosis,pathology,treatment method,prognosis,and so on were collected and analyzed.A reference of diagnosis and treatment is provided.

Objective To investigate status of the intravenous use of antibiotics in outpatients and emergency patients of a tertiary first-class hospital, and provide a reference for developing management measures in next step. Methods By a retrospective method,all the prescriptions using antibiotics by intravenous administration in outpatients and emergency department patients from a tertiary first-class hospital in 2013 were extracted from the hospital information system. The categories of antimicrobial agents,proportion of intravenous use of antimicrobial drugs,ranking of the antibiotic consumption sum and defined daily dose,and the top 10 clinical departments or wards intravenously using antimicrobial drugs were chosen to analyze. The data in 2014 were extracted as a comparison. Results Outpatients and emergency department patients respectively used 8 categories 31 kinds and 8 categories 30 kinds of intravenous antimicrobial drugs, with high consumption of cephalosporins and restricted antimicrobials such as sodium cefoxitin.Intravenous use of antimicrobial drug prescription proportion in emergency department is higher than that in outpatient department. Conclusion After intervention in 2014, antibiotic consumption is effectively controlled as compared that in 2013. But management should be strengthened and appropriate interventions should be taken to ensure the use of intravenous antibiotics in a safe,effective and economical manner.

HBV polymerase has intrinsic RNA-dependent reverse transcriptase, DNA-dependent DNA polymerase as well as RNaseH activity. Analysis of HBV polymerase has been hampered for many years due to the inability to express functional enzyme in a recombinant system. To obtain active polymerase at a high level, we have taken advantage of baculovirus expression system. The gene of HBV polymerase was amplified by PCR and cloned into pFastBac Dual to construct the recombinant plasmid pFastbac Dual-pol. The recombinant donor plasmid, pFastbac Dual-pol, was constructed by inserting HBV polymerase gene into EcoRI and PstI sites controlled by polyhedrin promoter. The recombinant donor plasmid was transformed into DH10Bac competent cells for transposition. Recombinant bacmid was constructed by inserting of the mini-Tn7 element from the donor plasmid into the mini-attTn7 attachment site on the bacmid. The recombinant bacmid DNA was isolated and transfected into the Sf9 cells to produce the recombinant virus, and healthy insect Sf9 cells were infected with the recombinant virus containing HBV polymerse gene to express the target protein. HBV polymerse expressed in insect cells was analyzed by SDS-PAGE. PCR results showed recombinant donor plasmid, pFastbac Dual-pol, was constructed successfully. The recombinant hepatitis B virus polymerase was expressed in insect cells at high level. The recombinant hepatitis B virus polymerase should facilitate the analysis of HBV polymerase biological characteristics, allow the investigation for new anti-HBV drugs specifically blocking HBV polymerase.