Objective:To clone and express mouse fibroblast growth factor eight(FGF 8) and investigate the function of it Methods:According to the published sequence of mouse fibroblast growth factor 8, a pair of special primers was designed Then the full length cDNA of FGF 8b (a predominant isoform of FGF 8) was isolated from the total RNA of the mouse embryo by means of reverse transcription PCR and subcloned into the yeast expression vector of pYEX4T 1 in correct direction It was identified with sequencing The recombinant plasmid was transformed into yeasts The fusion protein expressed was verified by using Western blot The activity of the protein was examined by MTT and 3H TdR incorporation Results:The cDNA fragment was about 600 bps, and proved to be "b" isoform of FGF 8 exactly This fusion protein, with molecular weight about 55 kD, was highly expressed in yeast system The data of MTT and 3H TdR incorporation in NIH3T3 cells were increased obviously by the protein, and decreased by the antagonist Conclusion:Fibroblast growth factor eight was cloned and expressed.The protein was found to be capable of stimulating the proliferation of NIH3T3 cells Moreover, the recombinant protein of the extracellular fragment of FGFR1 can antagonize the response of NIH3T3 to the recombinant protein

Objective: To express recombinant human soluble fibroblast growth factor receptor 1 ( sFGFR1) and study its antagonistic activity on FGF. Methods: Human sFGFR1 cDNA, isolated from human lung fibroblast cells with RT-PCR was confirmed by DNA sequencing and cloned into pYEX4T-1 yeast expression vector. The recombinant sFGFR1 was expressed in DY150 yeast cells and the product was identified by SDS-PAGE and Western blot. The activity of recombinant sFGFR1 was detected in N1H3T3 proliferation inhibition assay. Results: GSF-sFGFR1 fusion protein was expressed in yeast cells and was observed as a band of 60 Id) on a SDS- PAGE gel and by Western blot. The recombinant fusion protein was also found to be able to suppress FGF-induced proliferation of NIH3Th cells. Conclusion: Recombinant human GST-sFGFR1 fusion protein was expressed in yeast efficiently and showed natural biological activities.

OBJECTIVE:To explore the repairing effect and mechanism of Huazhuo jiedu yukui formula on the mucosal dam-age of rats with ulcerative colitis (UC). METHODS:72 rats were randomly divided into blank group,model group,mesalamine group(positive control,0.3 g/kg),Huazhuo jiedu yukui formula low-dose,medium-dose,high-dose groups(5,10,20 g/kg),12 in each group. Except for blank group,other groups were induced UC model. After 4 d of modeling,rats in administration groups received related medicines,ig,blank group and model group received normal saline,ig,once a day,for 2 weeks. After administra-tion,disease activity index(DAI)and mucosal damage index(CMDI)of rats were observed;IL-8,TNF-α contents in serum and cyclooxygenase 2 (COX-2) expression in colon tissue were detected. RESULTS:Compared with blank group,DAI and CMDI scores in model group were increased;IL-8,TNF-α contents in serum were increased;COX-2 expression in colon tissue was en-hanced,with statistical significances (P<0.05). Compared with model group,DAI and CMDI scores in model group were de-creased;IL-8,TNF-α contents in serum were reduced;COX-2 expression in colon tissue was weakened,with statistical signifi-cances (P<0.05). Compared with mesalamine group, the above-mentioned indicators in Huazhuo jiedu yukui formula medi-um-dose,high-dose groups changed more obviously (P<0.05),and effects of Huazhuo jiedu yukui formula showed significant dose-effect relationship. CONCLUSIONS:Huazhuo jiedu yukui formula can improve disease activity status of UC rats and reduce colon mucosal damage,and the effects of medium-dose and high-dose Huazhuo jiedu yukui formula were better than mesalamine, which may be related to reducing inflammatory reaction and inhibiting COX-2 expression in colon tissue.

Objective To investigate the predictive value of acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ) for nursing workload in emergency intensive care unit (EICU).Methods A retrospective cohort study was conducted. Critical patients except for trauma admitted to EICU of West China Hospital of Sichuan University from June 2012 to July 2016 were enrolled. APACHE Ⅱ score and therapeutic intervention scoring system (TISS-28) score were calculated in each case. The relationship between APACHE Ⅱ score and TISS-28 score was analyzed by correlation analysis and curve estimation.Results A total of 1148 patients were enrolled, including 689 male (60.0%), and age of (56±19) years old, APACHE Ⅱ score was 19 (13, 25), and TISS score was 19 (13, 25). The Spearman correlation coefficient between APACHE Ⅱ score and TISS-28 score was 0.849 (P < 0.001), and that among the linear model quadratic model, composite model, growth model, logarithm model, cubic model and exponential modelR2 were 0.669, 0.710, 0.792, 0.764, 0.765, 0.765, respectively (allP < 0.01), and the most suitable description for relationship between APACHE Ⅱ score and TISS-28 score curve was cubic model. The nursing workload was significantly increased when APACHE Ⅱ score ≥30, and it was decreased when APACHE Ⅱ score < 30.Conclusion APACHE Ⅱ was a useful score for predicting the nursing workload in EICU.

This study compared two techniques of artery anastomoses,renal artery to the ex- ternal iliac artery (ESA) and to the internal iliacartery (EEA) in renal transplantation.The operation time and the incidence of anastomotic stenosis was cut down significantely in ESA group.The blood flow in grafts has no difference in two groups with normal renal function.The utilization rate of grafts with multiple arteries was higher (94.4%) in ESA group.Back bench surgery for artery repain,cold ischemia time and renal damage were reduced in ESA group.

Objective To evaluate the feasibility of application of PBL method on otolaryngology education for medical undergraduates.Methods Totally 61 medical undergraduates of Class 2009 at Peking Union Medical College were enrolled into this study.There are 22 males and 39 females and aged 23-26 years old.They were grouped randomly into two groups: the PBL group (n=29) and the traditional teaching method group (n=32).Questionnaires were collected and statistical analyses were conducted.Results The scores of the evaluations for education of otolaryngology after teaching were much better than those before teaching in the PBL group in most aspects according to the questionnaires (P<0.05).In traditional teaching method group, there are only few aspects` scores much better than those before teaching.Comparing the two teaching methods shows, in most aspects, the scores of PBL group are much higher than those of the traditional teaching method group.ConclusionsCombination of PBL with other teaching method will be an ideal way for medical education in otolaryngology.

Objective To investigate the usage of antiepileptic drugs (AEDs),epileptic seizures,pregnancy outcomes and infant feeding practices among pregnant women with epilepsy,and provide scientific evidence for eugenics.Methods The clinical characteristics of 350 pregnant women with epilepsy from Shaanxi Provincial Epilepsy & Pregnancy Register from October 2012 to July 2016 were analyzed retrospectively.Results A total of 350 patients with 376 pregnancies were included in the final analysis.Among 376 pregnancies,272 pregnancies (72.3％) were under the treatment of AEDs.Of them,246 pregnancies (90.4％) were on monotherapy,26 (9.6％) on polytherapy.Only patients during 179 pregnancies (47.6％) took folic acid in the first trimester.Seizure frequency increased in patients during 123 pregnancies (32.7％),of whom patients during 43 pregnancies (35.0％) did not use AEDs.Although other patients during 80 pregnancies (65.0％) took AEDs,regular pharmacokinetic monitoring was absent.A total of 170 pregnancies (45.2％) attained seizure-free.Seizure frequency decreased in 42 pregnancies (11.2％) and remained unchanged in 24 pregnancies (6.4％).The rates of cesarean section and natural delivery were 43.8％ (158/361) and 56.2％ (203/361),respectively.Totally 4.5％ (17/376) pregnancies had adverse pregnancy outcome,and 1.9％ (7/376) had fetal malformations.The most common congenital malformation was heart malformation,which was observed in two offspring.One offspring developed dysostosis,one developed chromosome abnormality,and another developed cerebellum deformity.In addition,one twin developed conjoined deformities.Breastfeeding rate was 52.6％ (190/361).Conclusions In Shaanxi province,AEDs compliance,pharmacokinetic monitoring and usage rate of folic acid still need to be improved in pregnant women with epilepsy.Clinical trial registration Chinese Clinical Trial Registry,ChiCTR-OOC-16009277

Objective:To investigate the role of LncRNA ZBED3-AS1 in osteoblast proliferation and differentiation in osteoporotic rats through regulating miR-339-5p/Notch 1.Methods:The rat models of sham operation (Sham) group and model (Model) group were established, and the bone mineral density (BMD) of rats was examined. Rat osteoblasts were isolated and the expression of ZBED3-AS1 and miR-339-5p was detected by qRT-PCR. The osteoblasts of rats in Sham group and Model group were divided into different groups and transfected. CCK8, alizarin red (AR-S) staining and alkaline phosphatase (ALP) staining were used to detect the proliferation and differentiation ability of cells in each group. The distribution of ZBED3-AS1 in cells was determined by FISH assay. Double luciferase report confirmed the relationship between ZBED3-AS1 and miR-339-5p as well as miR-339-5p and Notch 1.Western blot was used to detect the expression of Notch pathway related factors.Results:The bone mineral density of femur in Model group was significantly lower than that in Sham group ( P=0.0057) . Compared with Sham group, the expression of ZBED3-AS1 in osteoblasts of Model group was lower, while that of miR-339-5p was higher (all P<0.05) . Overexpression of ZBED3-AS1 could promote the proliferation and differentiation of osteoblasts, while knockdown of ZBED3-AS1 could inhibit the proliferation and differentiation of osteoblasts (all P<0.05) . ZBED3-AS1 could regulate miR-339-5p as ceRNA (all P<0.05) . Overexpression of miR-339-5p can inhibit the proliferation and differentiation of osteoblasts, which can be partially saved by overexpression of ZBED3-AS1. Notch 1 was confirmed as a target of miR-339-5p, at the same time, interfering with the expression of ZBED3-AS1/miR-339-5p can affect the expression of Notch-1 protein, and the regulation of ZBED3-AS1/miR-339-5p on osteoblasts may be realized through Notch pathway. Conclusion:ZBED3-AS1 can be used as ceRNA to regulate miR-339-5p, and then affect the proliferation and differentiation of osteoblasts in osteoporotic rats.

BACKGROUND:Recent studies have found that bone marrow mesenchymal stem cels that culturedin vitro for a long time can naturaly differentiate into neural stem cels, which then differentiate into neurons and glial cels, thereby providing a new therapeutic thinking for Parkinson’s disease, sequela of cerebral infarction, cerebelar atrophy and brain dysplasia.
OBJECTIVE:To discuss the influence of neural stem cel transplantation on neurologic function of rats with cerebral hemorrhage at recovery stage and the relevant mechanism of action.
METHODS: Sixty male Sprague-Dawley rats were randomly divided into normal group (n=18), cerebral hemorrhage group (n=21) and transplantation group (n=21). Cerebral hemorrhage models were established in the latter two groups using VII type colagen enzyme induction method. At 21 days of modeling, rats in the transplantation group were injected neural stem cels via the tail vein, and those in the other two groups received the same volume of normal saline. At 7, 14, 21 days after cel transplantation, modified adhesive removal test (MST) was employed to evaluate the neurologic function of rats, and then the rats were kiled. RT-PCR was used to detect angiopoietin-1 mRNA expression in the bleeding tissues, and western blot assay was employed to measure tyrosine kinase receptor-2 protein expression.
RESULTS AND CONCLUSION:Compared with the normal group, the MST scores in the cerebral hemorrhage group and transplantation group were significantly decreased (P< 0.05). From the 7th day after transplantation, MST scores in the transplantation group were significantly higher than those in the cerebral hemorrhage group (P < 0.05). At 7, 14, 21 days after transplantation, expressions of angiopoietin-1 mRNA and tyrosine kinase receptor-2 protein were ranked as folows: transplantation group > cerebral hemorrhage group > normal group, and there was a significant difference among the three groups (P< 0.05). These findings indicate that neural stem cel transplantation can effectively promote the neurologic recovery of rats with cerebral hemorrhage at recovery stage, and the concrete mechanism may be related to the increase of angiopoietin-1 mRNA and tyrosine kinase receptor-2 protein in the bleeding tissues.

Objective o evaluate the association between IL-28B ( rs12979860 ) polymorphism andantiviraltherapeutic effectbydetecting the genotype of interleukin-28B( IL-28 B) in patients with hepatitis C ( HCV ) .Methods Of total 1153 HCV patients, 303 diagnosed with CHC had been treated with pegylated interferon plus ribavirin for 24-48 weeks.IL-28B ( rs12979860 ) was genotyped by two-color fluorescent TaqMan assay.Results Among 1153 patients, CC, CT and TT genotype frequencies of IL-28B rs12979860 are 83.26%, 16.22%and 0.52%respectively.The results of HCV genotypingof 580 in 1153 cases, the frequencies of 1b, 2a and their non-1b/2a type are 63.45%, 35.00%and 1.55%respectively;In 303 CHC patients with clear medical history, the proportion of SVR was71.98% in patients with CC genotype and 16.90%in those with either the CT or TT genotypes.Logistic regression model was adopted to analyze the association of rs12979860 with SVR while adjusting for age, gender, viral load and HCV GT factors.Populations carrying combined genotype ( CT +TT) are making it harder to get SVR compared with those with CC genotype (OR, 95%CI:11.10,5.35-23.04;P<0.000 1).The percentages of SVR in HVC patients with 1b and 2a genotypeare 48.02% and 81.19% respectively.there is a statistically significant difference between these subgroups (χ2 =30.639,P<0.000 1).Conclusion IL-28B rs12979860 genotype is closely related to SVR in CHCpatients.Patients with CC genotype have a higher virus sustained response rate than those carrying CT or TT genotype.The SNP , rs12979860, might be applied as a predictor of clinical antiviral efficacy in the furture.