0.05). 3. The morbidity of clinical and histological chorioamnionitis in ambroxol group (5.6% and 58.3%, respectively)was significantly lower than that in dexamethasone group (27.5% and 82.5%,respectively, P

Aim To construct human phage single-chain antibody library associated with esophageal cancer and to screen the specific scFv against Eca109 cells from the liberary. Methods Metastatic periesophageal lymph nodes of esophageal cancer patients were used as the B cells source, the total RNA of these B cells was extracted and prepared as the template of RT-PCR. First, we screened graticulely two pairs of primers of the heavy and light regions separately, then the V_H and V_L fragments were first amplified from the cDNA by the polymerase chain reaction (PCR). Second, the V_H-linker and V_L-linker were amplified from the V_H and V_L fragments. Last, the V_H-linker and V_L-linker were assembled into scFv gene fragments by SOE-PCR,and then Sfi I and Not I restriction site were inlet in it. ScFv gene was cloned into the pCANTAB-5E phagemid. Phagemids were introduced into E.coli TG1 by electrotransformation, followed by rescue of antibody-expressing phage using M13K07 helper-phage superinfection. Recombinant scFv phage library was constracted and PCR was used to identify the insert ratio of scFv antibodies library. Results of SfiI/Not I double digestion reaction positive insert clone were identified by 1.5% agarose gel electrophoresis. The phage library was panned with NHEEC and Eca109 cancer cells in suspension for four rounds. Strongly positive recombinant phage clones were used to infect E.coli HB2151. Expression of soluble scFv was induced by IPTG. Soluble scFv from periplasm were purified by affinity chromatography and identified by SDS-PAGE and Western blot. Cell ELISA , immunohistochemical staining and immunocytochemical staining were used to identify the activity of the soluble scFv. Results The result of agarose gel electrophoresis showed that total RNA of these B cells had two bands of 28 S and 18 S. The size of V_H fragment is about 450 bp,V_L fragment is about 350 bp and scFv is about 850 bp. The competence is 108 cfu??g-1 pUC18 DNA. Randomly digestive reac-tion showed that the positive insert ratio was 91.7% (22/24). After four rounds of panning, the fourth phage yield is 141 times as much as that of the first one. SDS-PAGE and Western blot showed that the MW of the soluble scFv was about 30 ku and the brand of 30 ku was stained. Immunohistochemical staining showed strong stainning of the tissue of esophageal cancer, but not the liver and gastric cancer tissue. Immunocytochemical staining showed significant staining of the esophageal cancer line Eca109. The result of cell ELISA assay revealed that soluble scFv had highly specific and could combined with Eca109 cells, but not with BGC-823 and NHEEC. Conclusion A human scFv phage display library associated with esophageal cancer has been constructed successfully and the specific scFv antibody against Eca109 has been identified from the liberary.

Totally, 102 general practitioners (GPs) from 40 community health-care service centers in a district of Beijing were investigated for their awareness of diagnosis, treatment and prevention for chronic hepatitis B and evaluating effectiveness of their training with self-designed questionnaire before and after professional training.Before training, higher awareness of knowledge about prevention for hepatitis B had been achieved in GPs, such as "hepatitis C is communicable" (78/102, 76.5% ), "hepatitis B can be prevented by protective antibody" ( 76/102, 74.5% ), whereas only 3.9% ( 4/102 ) of GPs knew indications of antiviral therapy for chronic hepatitis B, 4.9% (5/102) could describe complications of decompensated cirrhosis and 19.6% (20/102) knew methods for screening primary hepatocellular carcinoma.After training, their awareness increased significantly to 93.1% (95/102), 49.0% (50/102)and 86.3% (88/102), respectively (all P ＜0.01 ).Now, GPs' knowledge about diagnosis and treatment for hepatitis B is still insufficient at community health-care centers and professional training can improve their awareness.It is an important assurance for patients with chronic hepatitis B to be better managed in both of specialized hospitals and community health-care centers by enhancing professional training for GPs indiagnosis and treatment for hepatitis B.

Objective To investigate the effect of propofol on the cerebral injury induced by ketamine in neonatal rats. Methods Eighty 7-day-old SD rats of both sexes, weighing 12-20 g, were randomly divided into 4 groups (n = 20 each): normal saline (NS) group, ketamine-induced cerebral injury group (group K), propofol group (group P) and propofol combined with ketamine group (group PK). Group NS received intraperitoneal NS 1 ml. In groups K, P and PK, ketamine 70 mg/kg, propofol 70 mg/kg and propofol 70 mg/kg + ketamine 70 mg/kg were injected intraperitoneally once every 2 h for 3 times respectively. Ten rats in each group were selected and sacrificed at 24 h after emergence from anesthesia and the hippocampi obtained to determine the neuronal apoptosis (by TUNEL) and Bcl-2 and Bax protein expression(by immunohitochemistry). The apoptosis rate was calculated.The other 10 rats in each group were selected at 21 days after the intraperitoneal injection and the learning and memory functions (escape latency and frequency of crossing the original platform) were evaluated using Morris water maze. Results Compared with group NS, the apoptosis rate was significantly increased in group K, Bcl-2 protein expression was up-regulated in groups P and PK, and Bax protein expression was up-regulated, the escape latency was significantly prolonged and the frequency of crossing the original platform was significantly decreased in the other groups (P ＜ 0.05 .or 0.01 ). Compared with group K, the apoptosis rate was significantly decreased in group PK, Bax protein expression was down-regulated in group P, and Bcl-2 protein expression was up-regulated,the escape latency was significantly shortened and the frequency of crossing the original platform was significantlyincreased in groups P and PK ( P ＜ 0.05). Conclusion Propofol can reduce the cerebral injury induced by ketamine in neonatal rats, and the regulation of the Bcl-2 and Bax protein expression and inhibition of the neuronal apoptosis in hippocampus may be involved in the mechanism.

BACKGROUND:Previous experiments have prepared shape memory polymer based on D,L-poly(lactic acid). According to domestic technical requirements for biological y evaluating biomaterials and medical equipments, tissue-engineered grafts must be subjected to preclinical experiment for biosecurity and cytocompatibility evaluation.
OBJECTIVE:To observe the biosecurity of the shape memory polymer based on D,L-poly(lactic acid).
METHODS:(1) Bacterial endotoxin test:polymer extract, endotoxin working standard solution and checking
standard water were added into limulus reagent, respectively. (2) Sensitization test:Polymer extract+Freund’s complete adjuvant+physiological saline and Freund’s complete adjuvant+physiological saline were injected into the scapula of Kunming mice. After induction by intradermal injection, local induction and excitation, stimulate skin erythema and edema degree were observed in animals. (3) Acute toxicity test:Kunming mice received intraperitoneal injection of 100%, 50%, 25%polymer leaching solution and physiological saline, respectively. (4) 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay for cel proliferation:The direct method was that human umbilical cord vascular endothelial cel s were inoculated onto the polymer film, lactic acid and glass, respectively;the indirect method was that human umbilical cord vascular endothelial cel s were inoculated into the polymer leaching solution, acrylamide solution and 1640 culture solution.
RESULTS AND CONCLUSION:This shape memory polymer based on D,L-poly(lactic acid) is free of bacterial contamination in compliance with the biosecurity standards, and it has no al ergenic and toxicity but has good cytocompatibility.

Objective To explore the relationship between the obstetric factors defined by Robson Ten-Group Classification System (RTGCS) and cesarean section rate (CSR),and to evaluate the validity of predictive models established based on the RTGCS for cesarean section.Methods Clinical data of 41 295 cases delivered in Shanghai First Maternity and Infant Hospital from January 1,2012 to September 30,2014 were retrospectively collected.CSR of different groups categorized by the RTGCS was analyzed by descriptive epidemiological methods by three models.Model 1 was obstetrics factors (parity,history of cesarean section,number of fetus,mode of labor,fetal presentation and gestational age);model 2 was obstetrics factors (model 1) + demographic characteristics and severity (maternal age and with/without treatment in intensive care unit);model 3 was defined as model 2+ complications (placenta previa,placental abruption,chronic or gestational hypertension,preeclampsia,renal disease,or human immunodeficiency virus infection).The relationship between obstetrics factors classified by the RTGCS and CSR was analyzed by the logistic regression model (the potential confounders were controlled).The validity of predictive models was evaluated by analyzing the area under the receiver operative characteristic (ROC) curve.Results (1) There were statistical differences in total CSR and CSR before labor among those cases respectively grouped by age,medical insurance,parity,signal/multiple pregnancy,fetal presentation and gestational age (all P＜0.05).(2) The recruited cases were categorized into ten groups according to the RTGCS and among them,the second group (single,cephalic presentation,nulliparous,≥ 37 gestational weeks,induced labor or cesarean section before labor) accounted for most of the cesarean section cases (61.2％,11 217/18 322),followed by the fourth group (single,cephalic presentation,multiparous,no previous cesarean section,≥ 37 gestational weeks,induced labor or cesarean section before labor),which were 11.2％ (2 061/18 322).(3) Obstetrical parameters,including parity,number of fetus,mode of delivery,history of cesarean section,fetal presentation and gestational weeks at delivery were related to CSR,even after adjustment for maternal age and medical insurance,or obstetrical complications,or both above factors (all P＜0.05).(4) Areas under the ROC curves and the 95％ confidence intervals of model 1,2 and 3 were 0.867 (0.863-0.870),0.875 (0.872-0.878) and 0.881 (0.878-0.885),respectively (all P＜0.01).Conclusion Related variables of RTGCS highly correlate with CSR and could be used as promising indicators for prediction of cesarean section.The RTGCS is of great application value to those research subjects.

10.3969/j.issn.2095-4344.2013.25.017

Objective To investigate the pathogen characteristics of perforated appendicitis in children and the perioperative use of antimicrobials in order to provide evidence for the rational use of perioperative antibiotics. Methods The perioperative usage of antibiotics was analyzed to determine the reasonableness of antimicrobial use in children with perforated appendicitis who were discharged from July 2011 to August 2014,based on“guidelines of clinical use of antibiotics”and results of bacterial culture. Results Inflammatory secretions obtained from 126 children(126/ 149)were sent for examination and the examination rate was 84.56%.A total of 117 cases were found positive for cultured pathogens,and the detection positive rate was 92.86%.Three types of bacteria ranking the first three places were Escherichia coli,Pseudomonas aeruginosa and CitroBacter freundii.The utilization rate of antibacterial agents was 100.00%,with a dominant use of cephalosporins and nitrate imidazoles. Rational use of antimicrobial agents was found in 144 cases(accounting for 96.64%). Conclusion The major pathogen in perforated appendicitis is still Escherichia coli,which is highly sensitive to commonly used antibiotics,and drug-sensitivity testing results can help guide the treatment programs and antibiotics selection.

Purpose To explore the specific diagnostic markers of Sertoli cells and germ cells for testicular biopsy. Methods Normal testis from 3 patients who suffered from carcinoma of the prostate and treated with testis castration, and 15 testicular biopsy tissues were stained by EnVision two steps with WT-1, AR and Ki-67. Results The expression of WT-1 and AR protein were found in Sertoli cells from the 3 normal testis and 15 testis biopsy, the positive rate were all 100%, and non was positive in germ cells. The germ cell was positive for Ki-67, and the positive rate was 100%. Conclusions WT-1, AR and Ki-67 may be the specific diagnostic markers of Sertoli cells and germ cells for testicular biopsy, which may contribute to the diagnosis of testicular biopsy.

Objective:To investigate the off-label drug use in the outpatients of Zhengzhou children's hospital in 2013 to provide the evidence for improving the rational drug use in pediatrics. Methods:The prescriptions of outpatients were randomly selected from January to December in 2013. According to the drug instruction, the off-label drug uses were analyzed in the following aspects: sam-pling conditions, the kinds of off-label drug use, the incidence of off-label drug use in different age groups and drug categories. Re-sults:A total of 8 684 prescriptions with 16 344 medical records involving 410 kinds of drugs were analyzed, and the rate of off-label drug use was 70. 87% for prescription, 51. 21% for medical orders and 71. 95% for category, respectively. The primary types of off-label drug use were over applicable people (50. 83%), dosage(21. 00%) and frequency (19. 04%). The top 3 age groups with high-est off-label drug use rate were neonates (63. 27%), infants (56. 20%) and school-age (44. 98%). The top 5 drugs with highest off-label drug use rate were cardiovascular system medicines (99. 02%), anti-infective agents (64. 14%), vitamin and mineral drugs (62. 89%), hematological system medicines (59. 48%) and Chinese patent drugs (53. 43%). Conclusion: Off-label drug use in outpatients is prevalent. Physicians should master indications, usage and dosage described in instructions, select suitable dosage form, reduce off-label drug use and improve drug safety in pediatrics.