Objective To construct a recombinant adenovirus contained respiratory syncytial virus F gene fragment. Methods The F gene fragment was amplified by RT-PCR. Then the recombinant shuttle plasmid pShuttle-CMV/F was constructed. The iinearized pShuttle-CMV/F was transformed into BJ5183-AD-1 electroporation competent cells which contained plasmid pAdEasy-1. The obtained homologous recom-binant plasmid was named pAdEasy/F, and transfected into 293 cells. Then the recombinant adenovirus rAd/F was obtained, and it was tested by electron microscope, RT-PCR, Western blot and immuno-fluores-cence assay(IFA). The vires titer and genetic stability were also studied initially. Results The recombi-nant adenovirus rAd/F was constructed, which was visualized as typical adenovirus morphology under elec-tron microscope. The transcription and expression of RSV F gene fragment integrated in the rAd/F were con-finned by RT-PCR, Western blot and IFA. Conclusion The recombinant adenovirus rAd/F contained RSV F gene fragment is successfully constructed and lay an important foundation for the further vaccine study.

Objective To investigate the dual-direction regulation of Tongxie Yaofang on gastrointestinal movement by studying its effect on gastrointestinal movement of mice in various functional states. Methods Adopting diarrhea model induced by rhubarb,constipation model by compound diphenoxylate and normal mice,the effect of different doses of Tongxie Yaofang on loose stool quantity,degree,diameter,diarrhea rate,loose stool incidence rate,loose stool grade,diarrhea index of every group were observed. By esta blishing hyperfunction of small intestinal function in mice by neostigimine,inhibition of small intestinal function in mice by atropine sulfate and small intestinal propellant function in normal mice,the effect of Tongxie Yaofang on intestinal movement of mice were studied. Results Tongxie Yaofang could inhibit gastrointestinal movement of diarrhea model and hyperfunction of small intestinal function in mice by neostigimine. It could promote gastrointestinal movement of constipation model and inhibit small intestinal function in mice by atropine sulfate. But it had no obvious action on natural mice and its intestinal movement. Conclusions Tongxie Yaofang has dual-direction regulation on gastrointestinal movement of mice in various functional states,which has a good agreement with formula-syndrome in traditional Chinese medicine,conforms to treatment based on syndrome differentiation.

OBJECTIVE:To prepare in-situ gelling of hydrochloride sparfloxacin eye drops and establish its quality con-trol method.METHODS:The in-situ gelling of hydrochloride sparfloxacin eye drops were prepared with boric acid as buffer to regulate the pH and osmotic pressure and with sodium polymannuronate as peptizer.The content of hydrochloride sparflo-xacin was determined by ultraviolet spectrophotometry.The stability of the sustained-release eye drops was investigated as well.RESULTS:The sustained-release eye drops were colorless or yellowish limpid liquid with its test and identification results all in conformity with the related stipulation stated in Chinese Pharmacopeia(2005 edition).There was a good linear relationship within the concentration range of 3.0～8.0 ?g?mL-1 for hydrochloride sparfloxacin,and its average recovery was 99.82%(RSD=0.236%,n=6).There was no significant change for indexes in the accelerate test and sample test,yet the temperature and lighting did have slight impact on the viscosity,osmotic pressure and pH of the solution.CONCLUSION:The preparative method is simple and feasible and the quality of in-situ gelling of hydrochloride sparfloxacin eye drops is stable and controllable.

1-Alkyl-2-acetylglycerophosphocholine Esterase ; blood ; genetics ; Adolescent ; Asthma ; blood ; enzymology ; genetics ; Child ; Child, Preschool ; Female ; Gene Expression Regulation, Enzymologic ; Genotype ; Humans ; Infant ; Male ; Mutation ; Polymerase Chain Reaction

Objective To investigate the existence of pulmonary vascular remodeling after left pneumonectomy in rats and the role of hypoxia inducible factor-lα( HIF-1α) and vascular endothelial growth factor ( VEGF) in pulmonary vascular remodeling.Methods Twenty-four healthy male Sprague-Dawley rats were randomly divided into experimental and control groups, 12 in each group.The rat models of pulmonary vascular remodeling were created by open-chest left pneumonectomy.After 12 weeks of feeding, the mean pulmonary artery pressure ( mPAP) and partial pressure of arterial oxygen ( PaO2 ) of each rat were measured.The ultrastructure of small arteries in the lung specimens were examined by e-lectron microscopy.Muscularized degree of three kinds of small pulmonary vessels ( muscularized artery MA, partially mus-cularized artery PMA, and non-muscularized artery NMA) were observed by light microscopy, and the percentage of each kind of pulmonary arteries ( MA%, PMA%, NMA%) were calculated.Arterial external diameter, media thickness of ves-sel ( MTV) , total vascular area, media area of vessel ( MAV) , MTV%and MAV%were calculated as indicators of pul-monary vascular remodeling.The expressions of HIF-1αand VEGF in artery were detected by immunohistochemistry.Re-sults The values of mPAP, MA%, PMA%, MTV, MAV, MTV% and MAV% in the experimental group were signifi-cantly higher than those in the control group (P<0.01), but the value of PaO2 and NMA%were significantly lower than those in the control group (P<0.01).The IOD value of HIF-1αand VEGF expressed in the pulmonary arterial wall of the experimental group were 26.47 ±4.16 and 42.04 ±3.79, respectively, significantly higher than those in the control group (6.12 ±2.14 and 11.53 ±2.29, P<0.01).Linear correlation analysis showed that the expression of HIF-1αand VEGF was positively correlated with MTV% and MAV%, negatively correlated with PaO2 , and the HIF-1αexpression was posi-tively correlated with VEGF expression.Conclusions A rat model of pulmonary vascular remodeling can be successfully established by left pneumonectomy.Hypoxia is a key factor in the development of pulmonary vascular remodeling, HIF-1αand VEGF may play an important role in its pathogenesis.

Objective To investigate the microRNAs(miRNAs)expression profiles in the bowels of patients with Hirschsprung disease(HSCR),and to explore the role of differentially expressed miRNAs in the pathogenesis of HSCR. Methods Twenty - seven HSCR tissues,including spastic segments and distending segments,were obtained from patients with HSCR during operation. Then miRNA microarrays were used to investigate the miRNA expression profiles in 6 HSCR specimens. Bioinformatics software was used to predict target genes of miRNA. Three miRNAs (miR - 145 - 3p,miR - 4505 and miR - 1260a)were chosen and quantificational real - time(qRT)- PCR was per-formed to verify the different expression of those three miRNAs in 27 HSCR tissues. Results The expression of 26 miRNAs in an aganglionic colon segment was found to be more than two fold greater than that in ganglionic segment tis-sues,including 19 up - regulated miRNAs and 7 down - regulated miRNAs in patients with HSCR(all P ﹤ 0. 05). Tar-get genes of miRNAs were found,such as SOX10,RET,L1CAM. qRT - PCR showed the expression of miR - 145 - 3p (1. 42 ± 0. 42,aganglionic segment vs 0. 90 ± 0. 31,ganglionic segment)and miR - 4505(1. 30 ± 0. 30,aganglionic segment vs 0. 76 ± 0. 22,ganglionic segment)displayed a statistical difference between groups(all P ﹤ 0. 001). Be-sides,the expressions of miR - 145 - 3p(1. 53 ± 0. 46,long - segment type vs 1. 16 ± 0. 12,short - segment type)and miR - 4505(1. 42 ± 0. 26,long - segment type vs 1. 00 ± 0. 16,short - segment type)showed a statistical difference be-tween different types(all P ﹤ 0. 001),but miR - 1260a(1. 11 ± 0. 25,aganglionic segment vs 0. 99 ± 0. 21,ganglionic segment)did not show differential expression between different groups(P = 0. 064). Conclusions Abnormal expression of miRNAs was found in HSCR spastic segments,suggesting that miRNAs may be involved in the pathogenesis of HSCR.

Objective To investigate the incidence of contrast induced nephropathy (CIN) among different patient groups after contrast agent injection.Methods A total of 1243 patients were included in this study (male =694,female =549).The SCr level one week before and 72 hours after the CT examination and the incidence of CIN were recorded and comparison was made among groups according to sex,age,body mass index (BMI),the history of high blood pressure (HBP),diabetes mellitus (DM),chronic kidney disease (CKD),chronic heart failure (CHF),tumor,nephrotoxicity drug (NTD) usage.The frequency,type,dose and injection velocity of the contrast media(CM)were also recorded.Multivariate predictors of CIN were identified by Logistic regression using step-wise selection with entry and exit criteria of P ＜0.10,results were tabulated as odds ratios (OR) with 95％ confidence intervals (CI).Results Among 1243 consecutive patients,the incidence of CIN was 5.5％ (68/1243).Patients with a history of HBP,DM,CHF,CKD or tumor presented with higher incidence of CIN than that of controls (5.9％,51/868 vs.4.5％,17/375).CIN developed in 9 of 203 patients (4.4％,9/203) with CKD and in 59 of 1040 patients (5.7％,59/1040)without CKD.There was no significant difference between the two groups(x2 =0.51,P =0.30).In CKD (-) group,the incidence of CIN was higher in females,patients with DM and patients using LOCM than those of males,DM (-) and using low osmolality contrast medium (IOCM) (P ＜ 0.05),but there was no statistical significance in CKD (+) group.Logistic regression analysis showed that women,age ≥ 75 years,DM,LOCM,NTD,tumor,the time of using CM more than once per month were the most significant predictors of CIN (OR ＞ 1).Conclusion Women,age ≥ 75 years,LOCM,NTD,tumor,and the frequency of using CM more than once per month were more likely to develop CIN.

Objective To investigate the survival benefit of patients with esophageal squamous cell carcinomas who recieved postoperative chemotherapy of tegafur combined with FT-207 radiotherapy.Methods Data from 126 patients with esophageal squamous were analyzed who undergone radical resection from January 2008 to December 2010,consisting of 92 patients underwent surgery alone and 34 patients took oxaliplatin combined with tegafur postoperatively (oxaliplatin 130 g/m2,d1,tegafur 500 mg/m2,d1-5,21 d for a cycle,every patient received at least two cycles).Results The 1-year survival rate and 3-year survival rate was 93.5 ％ and 75.0 ％ in the surgery alone group,was 97.1％ and 76.2 ％ in the surgery plus postoperative chemotherapy (P =0.590).There was no significant differences in survival in the two groups,even with Coxregression multivariate analysis and with stratifications of gender,age,tumor location,degree of cell differentiation,invasion depth,lymphatic metastasis and TNM stage.Conclusion Postoperative adjuvant chemotherapy with oxaliplatin combined with tegafur has not significantly improve the survival rate of patients with esophageal squamous cell carcinomas undergoing radical resection.It is still lack of an effective project for postoperative adjuvant chemotherapy for esophageal cancer,and it is necessary for further study.

Objective To evaluate the clinical efficacy and toxicities of radical intensity-modulated radiotherapy (IMRT) combined with reduction in dose of prophylactic irradiation in the treatment of stage Ⅲ small cell lung cancer (SCLC).Methods A retrospective analysis was performed on the clinical data of 40 patients with stage Ⅲ SCLC who were admitted from January 2010 to August 2012.The prescribed dose was 60 Gy in 30 fractions to the primary gross tumor volume and was 54 Gy in 30 fractions to the planning target volume.All patients received induction chemotherapy,31 patients received adjuvant chemotherapy,and 22 patients received concurrent chemoradiotherapy;the platinum-based chemotherapy combined with etoposide or teniposide was adopted.Prophylactic cranial irradiation (25 Gy in 10 fractions) was administered to 17 patients.The short-term tumor response was evaluated by RECIST 1.0,and radiation-related toxicities were assessed by CTCAE 4.0.Overall survival (OS),local recurrence-free survival (LRFS),and progression-free survival (PFS) were calculated by Kaplan-Meier method.Results The short-term tumor response rate was 98％.The follow-up rate was 100％.Twenty-two patients were followed up for at least 2 years.The 1-and 2-year OS rates were 84％ and 48％,respectively; the LRFS rates were 89％ and 85％,respectively; the PFS rates were 61％ and 41％,respectively.Grade 0-1 radiation-related pneumonia was observed in 65％(26/40) of all patients,grade 2 in 25％ (10/40),grade 3 in 5％ (2/40),and grade 5 in 5％ (2/40).Grade 0-1 radiation-related esophagitis was observed in 53％ (21/40) of all patients,grade 2 in 43％ (17/40),and grade 3 in 5 ％ (2/40).Conclusions Preliminary results from this study suggested that IMRT combined with reduction in dose of prophylactic irradiation is safe and effective in patients with stage Ⅲ SCLC and is worth further evaluation in a large,prospective,randomized study.