Objective:To construct the helper virus VCSM13N1 with amber mutation,so as to provide a platform for the selective infective phage(SIP) technique.Methods: A pair of primers with amber stop codon were synthesized and introduced into N1 region of helper virus VCSM13 protein by overlapping extension PCR,and the products were identified by ristriction enzyme digestion and sequencing.The switch on/off efficiency of amber codon was detected with E.coli XL1-Blue and HB2151.The feasibility of using the amber mutant phage as helper virus was determined with anti-HBsAg phage antibody.Comprehensive analysis was also made on the performance of the amber mutant phage.Results: The titer of mutant phage was 2.5?10~(11) when detected with E.coli XL1-Blue,comparable with that of wild phage.The titer of mutant phage was at 10~6 level when E.coli HB2151 was used,being 10 000 folds lower than that of wild phage,when the mutant phages from HB2151 were used to infect E.coli XL1-Blue,the titer peaked at 5?10~(9),being 50 folds lower than that of wild phage.The activity of phage antibody prepared with mutant virus was comparable with the phage antibody prepared with wild phage.Conclusion: The constructed amber mutant phage can be used as helper virus,and it can switch off efficiently in non-suppressing strain HB2151.

Selectively infective phage (SIP) technology was developed for screening interacting protein-protein pairs. The in vivo SIP strategy would in principle be suitable for??library-library??selections, and the co-packaged polyphage may be a suitable approach. In order to construct the antigen expression vector which can be co-packaged into polyphage with phage displaying vectors, plasmid TG10 was chosen as the basic vector which is compatible with antibody display vector. The interval sequence of phage genome was amplified with PCR and cloned into TG10 to provide the packaging signal. It was named pTMI and it can be packaged into phage particles in 1011 level. The N1N2 region of gene ￠? was amplified and cloned into pTMI under the control of lac promoter to give pTMIN. Promoter trc was synthesized and replaced the lac promoter to give pTTMIN which permits the fusion expression of antigen with N1N2. To test its ability for fusion expression, gene code for ten-peptide of c-myc was synthesized and inserted into pTTMIN downstream to N1N2. After induction expression, the results of ELISA and SDS-PAGE showed that it has been expressed successfully. When pTTMIN was transfected into cell carrying antibody display vector p3MHHB3, it was copackaged into phage particles in 0.3% to 55% after rescuing with helper phage VCSM13.From the results it can concluded that the antigen expression vector was constructed successfully and it can be used for library-library screening in theory.

Objective To construct a packaged bacteria strain and, with it, to prepare gene III-defective helper phage. Methods Whole gene III and chloramphenicol-resistant gene were amplified and joined to form a fragment flanking 36bp homologous sequence of hisBCD using overlapping extending PCR, and it was then integrated into chromosome of E. coli XL1-Blue to replace the hisBCD gene using Red recombination system. The recombinant strain was identified with chloramphenicol-resistant screening, PCR and histidine-phenotype analysis, and named as XL-pⅢ. The gene III-deleted genome of phage was constructed with PCR and transfected into the recombinant strain XL-pⅢ to prepare the defective helper phage. It was quantified by infecting E. coli XL1-Blue and formed colonies were counted in kanamycin plate. Results The recombinant strain could grow in chloramphenicol-resistant plate, but couldn't grow in M9 medium without histidine, implying that it had lost its histidine phenotype. The aimed gene fragment could be amplified as designed. The constructed recombinant was named as XL-pⅢ. The prepared defective helper phage could infect E. coli XL1-Blue only once and the CFU was 3.7?108. Conclusions The packaged strain XL-pⅢ is successfully constructed and used to prepare the gene III-deleted helper phage. It is expected to be used in SIP system.

Objective To explore the effect of spironolactone on vascular remodeling of injuried rabbit artery. Methods 20 male New Zealand rabbits were randomly divided into two groups. The rabbit models of iliac artery angioplasty were set up by BC. After injury, control group was fed with standard forage, and experimental group was fed with standard forage and spironolactone(20mg?kg~ -1?d~ -1). The rabbits were killed four weeks after injury. The injuried segments of iliac artery were removed, and processed for histological and morphological analysis by imumohistochemistry and TD 2000 analysis system of pathology image. Results AUIEM and AUEEM in the experimental group significantly increased as compared with the control group(P0.05). Conclusion Spironolactone could improve vascular remodeling after angioploasty of iliac artery in rabbits.

Objective To explore the effect of inflammation and autoimmunity in peripartum cardiomyopathy(PPCM). MethodsA total of82 PPCM patients and 100 normal delivery patients were randomly selected and conducted epidemiological survey.High-sensitivity Creaction protein(hs-CRP),troponin I,human antimyocardial antibody IgG(AMA-IgG),Coxsackie B virus IgG(CBV-IgG)and adenovirus antibody IgG(ADV-IgG)were detected with ELISA. ResultsCompared with control group,PPCM patients had older age,higher pressure,higher proportion of cesarean section and infection.The levels of serum hs-CRP,cTNI,and leucocyte were markedly higher in PPCM patients compared with control.The positive proportion of AMA-IgG and CBV-IgG was significantly increased(P<0.01)in PPCM patients compared with the control.Logistic regression showed that infection(OR=2.87,95%CI 1.15-5.24),increased hs-CRP(OR=1.86,95%CI 1.08-4.02)and positive AMA-IgG(OR=2.68,95%CI 1.19-4.85)were independent risk factors for PPCM. ConclusionsInflammation and autoimmunity play an important role in peripartum cardiomyopathy.

Objective To explore the risk factors of peripartum cardiomyopathy (PPCM). Methods A total of 82 PPCM and 100 normal delivery females were randomly recruited in the current study. Echocardiographic, cTNI,high sensitive C-reaction protein(hs-CRP) ,NT-proBNP were measured. Fifty-two patients were followed up for a mean of 6 months. Results The PPCM patients (29. 5 ± 6. 4) yrs were older than the controls (25. 2 ± 5. 8) yrs (P<0.01) . Compared to the controls, the PPCM patients showed higher blood pressure (146.9/98. 8 mm Hg v. s. 130. 2/80. 1 mm Hg, P < 0. 01) , higher proportion of cesarean section (65. 9% v. s. 51. 0% , P < 0. 05) and complicated infection(75. 6% v. s. 10. 0% ,P <0. 01). The level of leucocyte(11.0 × 109/L) ,cTNI(0. 17 μg/L), hs-CRP(28. 2 mg/L)and NT-proBNP(650. 1 ng/L) were significantly higher in the PPCM patients compared with control(8. 8 × 109/L,0. 06μg/L,6. 2 mg/L and 110. 5 ng/L,P <0. 01). Hs-CRP was positively related with NT-proBNP(r = 0. 67, P < 0. 01). After average of 6 months of treatment, left ventricular ejection fraction enhanced from 32. 2% to 50. 6%. NT-proBNP significantly declined from 650. 1 ng/L to 225.6 ng/L(P <0. 01). Multiple linear regression analysis showed that diastolic pressure (P < 0. 05), LVED (P < 0. 05) and hs-CRP (P < 0. 05) were the independent predictors for declined NT-proBNP. Conclusions Inflammatory plays an important role in the development of peripartum cardiomyopathy.

Objective To discuss the safety and effectiveness of transurethral resection of the prostate (TURP) on large-size (≥ 80 ml) benign prostatic hyperplasia (BPH).Methods Retrospective analysis of 958 BPH patients in Southwest Hospital during January 2010 to January 2013 was conducted.The patients were grouped into ≥80 ml prostate group (Group A) and ＜80 ml prostate group (Group B) according to the volume of prostate.Comparison was made between the 2 groups on the safety and effectiveness of TURP.Results There were 276 patients in Group A and 682 in Group B.No significant differences were shown in average age and preoperative American society of anesthesiology score of Group A and B.Compared with Group B,decrement in hemoglobin level and blood Na+ concentration of Group A was more significant (P＜0.01).There were more prostate tissues excised and duration of the operations was longer (P＜0.01).No significant difference was observed in peri-operative complications graded by the modified Clavien classification system,catheter durations and durations of hospital stay between the 2 groups (P＞0.05).At 6 months after the surgery,average maximum urinary flow rate (Qmax) increased from 5.9±2.9 ml/s to 17.1±8.2 ml/s for Group A and 6.1±3.0 ml/s to 17.5±6.4 ml/s for Group B,both groups showed significant increase in Qmax after surgery(P＜0.01).Six months after surgery,international prostate symptom score (IPSS) of Group A decreased from 23.7±6.1 to 5.9±4.9 while IPSS of Group B decreased from 23.1±5.5 to 6.2±4.4,both groups showed a significant decrease (P＜0.01).No significant difference was shown in IPSS,quality of life,Qmax,postvoid residual urine volume and occurrence rate of long-term complications after 6 months between the 2 groups (P＞0.05).Conclusion TURP is as safe and effective in treating large-size BPH as treating medium and small-size BPH.

Objective To study the preparation method and analytical technique of hyperoside from Flos Abelmoschus manihot.Methods Hyperoside was isolated and purified by solvent extract and chromatography, whose structure was determined by 1H-NMR and 13C-NMR. The purity was analyzed by TLC and HPLC.Results The TLC showed that the hyperoside had no impurity spot. The HPLC indicated that the purity reached more than 98.5%.Conclusions The mothod of isolation and purification for hyperoside reported in this paper was simple and economical.

Objective To investigate the association between the osteogenic differentiation in the soft tissue lump and the clinicopathological characteristics of osteosarcoma patients. Methods We retrospectively reviewed the clinical data of conventional osteosarcoma patients with soft tissue lumps, including Enneking stages, chemotherapy sensitivity, overall survival and post-metastatic survival time. The ossification level in soft tissue lumps was assessed by imaging and the proportion of osteoid matrix was assessed by pathological examination. Results A total of 189 cases were included in this study. In patients with Enneking IIIB, non-osteoblastic, partially osteoblastic and osteoblastic types accounted for 30.2%, 9.6% and 6.3%, respectively. Non-osteoblastic osteosarcoma patients had a higher rate of initial metastasis (P < 0.05); Chemotherapy efficiency of non-osteoblastic, partially osteoblastic and osteoblastic types were 60.5%, 59.6% and 31.3%, respectively. The osteoblastic osteosarcoma held the worst rate of chemotherapy sensitivity (P < 0.05). The overall survival of non-osteoblastic osteosarcoma was shorter than those of partially osteoblastic and osteoblastic types (P < 0.05). Post-metastatic survival time of osteoblastic osteosarcoma was longer than that of non-osteoblastic osteosarcoma (P=0.078). Conclusion For conventional osteosarcoma, the osteogenesis level in soft tissue lumps is related to the surgical stage, chemotherapy sensitivity and prognosis of tumors, which may provide guidance for the individual decision regarding chemotherapy and surgery timing on patients.

Objective To explore the association of TBX 5 polymorphisms and environmental exposure index with susceptibility to oral cancer. Methods A case?control study was conducted to collect 300 oral cancer patients hospitalized in the Department of Oral and Maxillofacial Surgery, the First Affiliated Hospital of Fujian Medical University from September 2010 to December 2016. A total of 445 non?tumor patients were selected as the control group. Questionnaires were used to collect the information of all subjects and 5 ml peripheral blood was collected to detect single nucleotide polymorphisms (SNPs) of the rs10492336 locus of TBX5 gene. According to the environmental exposure index score, subjects were divided into two groups, low risk group (0-2.31) and high risk group (2.32-11.76). To analyze the association of TBX5 gene rs10492336 SNPs, environmental exposure index and oral cancer and its interactions. Results The age of all subjects in the case group and control group were (56.19±13.10) years and (54.56± 12.48) years old. Compared with CC genotype, the OR (95%CI) values of the co?dominant genetic model AC genotype and the dominant genetic model AC+AA genotype were 0.69 (0.49-0.98) and 0.70 (0.51-0.97), respectively. Compared with the low risk group, the OR (95%CI) risk of oral cancer in the high risk group was 3.72 (2.55-5.43). The results of gene?environment interaction analysis showed that compared with the group with CC genotype and high risk of environmental exposure index, the OR (95%CI) value of oral cancer in the group with AC+AA genotype and low risk of environmental exposure index was 0.18(0.10-0.31). Furthermore there was a multiplicative interaction between rs10492336 SNPs and environmental exposure index (β=-0.405, P<0.001). Conclusion This study suggests that the TBX 5 gene rs10492336 SNPs and environmental exposure index were associated with oral cancer. And there was a multiplication interaction between rs10492336 SNPs and environmental exposure index.