Objective To explore effect of hyperoxia and amygdalin on surfactant associated protein messenger RNA levels of alveolar epithelial cells of premature rat lung. Methods Type Ⅱ alveolar epithelial cells (AECⅡ) were gained by primary culture from 19-days fetal rat lung. After purified,AECⅡ were randomly assigned to four groups and exposed to air or hyperoxia: air group (group Ⅰ),hyperoxia group (group Ⅱ),air plus amygdalin group (group Ⅲ),hyperoxia plus amygdalin group (group Ⅳ),Groups Ⅱ、Ⅳ were flushed the flake with 95% oxygen-5% CO 2 at 3 L/min for 10 min,then sealed and cultured for 24 hours. Groups Ⅲ,Ⅳ were added 200 ?mol/L of amygdalin at the same time. All groups were in CO 2 culture chamber (37 ℃,5% CO 2) for 24 hours,cells were harvested and extracted for total RNA by Trizol reagent. mRNA levels of SP were measured by reverse transcription polymerase chain reaction (RT-PCR). Results SP mRNA levels were significantly decreased in groupⅡ compared to groupⅠ( P

Objective To observe the effects of the intervention of Didang Decoction at different times on changes of AMPK signaling pathway related factors in macrovascular endotheliocytes of diabetic rats; To discuss the mechanism of mitochondria energy metabolism regulating the AMPK signaling pathway for macrovascular endothelial defense function. Methods Injection of STZ into the caudal vein and administration of high fat diet wer used to generate diabetic rat model. All rats were randomly divided into the following 7 groups: control, model, metformin, simvastatin, early-, middle-, and late-stage Didang Decoction group. Western blot was used to detect the expressions of APMKα1 and PGC-1α in rat aortic endothelial cells. Changes in the intracellular AMP and ATP levels were detected by ELISA. Real time fluorescence quantitative PCR was used to detected mRNA expressions of Caspase-3, eNOS, and Bcl-2 in tissue of thoracic aorta. Results Compared with the model group, the expressions of AMPKα1 and PGC-1α in the early-stage and middle-stage Didang Decoction group and simvastatin group increased (P<0.05); the gene expressions of Bcl-2, and eNOS significantly increased in the early-stage Didang Decoction group and simvastatin group (P<0.05), while the expressions of Caspase-3 decreased significantly (P<0.05). The expression of ATP increased significantly and the expression of AMP decreased significantly in the early-stage Didang Decoction group and simvastatin group (P<0.05), and the best effects were shown in the early-stage Didang Decoction group. Conclusion Early intervention of Didang Decoction can enhance energy metabolism in the mitochondria of macrovascular endothelial cells by regulating the AMPK signaling pathway, and then plays a role in strengthening the defense function of macrovascular endothelial cells.

Objective To evaluate the surgical technique and procedures of transumbilieal breast augmentation with an inverse U-shape incision and insertion of saline-filled breast implants. Methods With specialized instruments, the subcutaneous tunnel was undermined to the inframammary crease after inverse U incision made along the umbilical border. The subpectoral space was dissected with a dissector after the advancement of obturator across the inframammary crease. The expander was inserted into the subpectoral pocket to be filled to obtain the high degree of symmetry of the breasts. Finally, the previous-ly prepared implants were inserted after the expander deflated and removed. Results All patients gained satisfactory results with no complications, such as hematoma, infection, implant deflation and Baker Ⅲ or Ⅳ capsular contracture. Conclusions Breast augmentation could be performed through umbilical inci-sion with the usage of saline-filled breast implant. This is an alternative of incisions in breast augmenta-tion.

Objective:To construct a human Fab fragment phage display library and provide a platform for human antibody preparation.Methods:Peripheral blood lymphocytes were collected from healthy donor.The heavy chain Fd fragment and light chain of human immunoglobulin's genes were amplified by RT-PCR,and then cloned into phagemid pComb3XSS to generate human phage antibody library.Cutting with endonucleases such as SacⅠ,XbaⅠ,XhoⅠand SpeⅠto identify the insertion of the light chain or heavy chain Fd genes.IL-2 and digoxin as the antigen was used to scan the phage antibody library.Results:A phage antibody library of Fab had 8.4?107 members and it's recombinant rate was 70%.Through DNA sequencing of one positive clone,it was showed that its heavy chain belonged to IgG subvariety and its light chain to ? family.Conclusion:The success of constructing a nave human phage antibody library proves the useful of phage display system in human antibody preparation,and it can be used to select,purify and express of amylin Fab antibody.

Objective:To obtain antibodies against amylin from a 'naive' human Fab fragment antibody phage diasplay library and to analyze the specificity of antigen binding activity.Methods:Panning and screening Fab antibody from the antibody library,the positive clones with well reactivity to amylin were selected after five times selection of 'adsorption-elution-enrichment'.Then the plasmid DNA which was extracted from the clones,was digested with Spe Ⅰ and Nhe Ⅰ to delete gⅢ (about 660 bp).The digested 47 000 bp DNA which was purified after separation of bands from agarose gel was ligated with T4-DNA ligase.The constructed expressing phagemids were transformed to the BL21(DE3)pLysS,soluble Fab was expressed in it by the induction of IPTG and its characteristics and specificity were determined by ELISA and Western blot.Results:Soluble Fab antibodies were expressed in E.coli.According with molecular weight of IgG Fab,protein band of about 47 kD was shown by SDS-PAGE.Western blot using the goat anti human IgG-HRP showed their binding activities.ELISA showed their specificity with amylin antigens and they did not react with bovine serum albumin.Conclusion:The high level expression and identification of the soluble human anti- amylin Fab fragment antibodies has been obtained successfully,which lays a solid foundation for further researching about the biological and pathological activities of amylin.

Objective To evaluate short-term clinical effects of modified total pelvic floor reconstruction surgery in treatment of severe pelvic organ prolapse.Methods Thirty-nine cases with severe pelvic organ prolapse including vault prolapse diagnosed by pelvic organ prolapse quantification (POP-Q) system received modified total pelvic floor reconstruction surgery.Clinical parameters associated peri-operative period and 12 months after surgery and complications were analyzed Results Median operation time was 70 minutes (30-240 minutes),median blood loss was 100 ml (10-200 ml).Seventy-seven percent (30/39) patients were able to micturate spontaneously in the next morning after surgery with postvoid residual volume less than 100 ml (0-650 ml).No severe intra-operative complications were recorded and the rate of postoperative morbidity was 20% (8/39 ).Median post-operative hospital stay was 4 days (1-11 days).The patients were followed up at median 24 months(13-29 months).According to POP-Q system evaluation,the successful rate of operation reached 100% .Two cases (5%,2/39) were recorded as symptomatic recurrence which manifested as posterior wall prolapse within 24 months after operation.During follow-up,8% (3/39) patients were found to have erosion within 7 months after surgery,and urgent urinary incontinence was observed in 5% (2/39) cases,while constipation occurred in 8% (3/39) cases.The most remarkable complication was dyspareunia (36%,5/14); while 50% (7/14) experienced better sexual life after surgery.Conclusions Modified total pelvic reconstruction is a safe,efficient and micro-invasive surgery in treatment of severe pelvic organ prolapse.However,its influence on post-operative sexual life should be concerned.

Objective To investigate the function of helper T lymphocyte cell(Th)1/Th2 cytokine in food allergy development.Methods A total 110 Balb/c mice were randomly divided into 2 groups:control group(40 mice)and food allergy group(70 mice).Food allergy animal models were established by ovalbumin,performed by skin prick test;in positive reaction mice,serum specific IgE,IL-4 and IFN-? were measured by enzyme linked immuosorbent assay(ELISA),and intestinal pathology were performed,and the mRNA expressions of IL-10,TGF-? in intestinal were measured by real-time PCR assay.Results In food allergy group,the mRNA expression of IL-10,TGF? in intestinal decreased(P

Objective To evaluate the effects of tofacitinib on liver function in rheumatoid arthritis (RA) patients.Methods Literature search was performed in databases including PubMed,Cochrane Library,China National Knowledge Infrastructure and Wanfang to identify randomized controlled trials about RA treated with tofacitinib.The retrieval time was up to August 2016.Meta-analysis was conducted by Revman 5.5 software.Results A total of 7 studies were included,involving 2 965 patients.The results of Meta-analysis revealed that the incidence of alanine transaminase (ALT)＞1 upper limit of normal (ULN) in patients receiving both 5 mg and 10 mg bid tofacitinib was significantly higher than placebo [5 mg bid tofacitinib:RR=1.48,95％CI (1.20,1.82),P=0.000 2;10 mg bid tofacitinib:RR=1.67,95％CI (1.37,2.05),P＜0.01];there was no significant difference in the incidence of ALT＞3 ULN [5 mg bid tofacitinib:RR=1.81,95％CI (0.57,5.79),P=0.32;10 mg bid tofacitinib:RR=1.36,95％CI (0.57,5.25),P=0.49];the incidence of aspartate transaminase (AST)＞1 ULN was significantly higher than placebo [5 mg bid tofacitinib:RR=1.59,95％CI (1.25,2.03),P=0.000 2;10 mg bid tofacitinib:RR=1.90,95％CI(1.50,2.40),P＜0.01],there was no significant difference in the incidence of AST＞3 ULN [5 mg bid tofacitinib:RR=1.17,95％CI (0.27,5.17),P=0.83;10 mg bid tofacitinib:RR=0.95,95％CI (0.26,3.44),P=0.94].Conclusion Tofacitinib slightly increases ALT and AST in patient with RA.Due to the limited sources and lack of domestic studies,more randomized controlled trials are still needed to verify the above conclusion.

ObjectiveTo explore the relationship between the changes of neuro-electrophysiology and prognosis in children with Guillain-Barr? syndrome(GBS).MethodsThirty-eight children with GBS were divided into group A(rapid recovery,n=16) and group B(slow recovery,n=22) according to the time required for podosoma motor function recovery,at the same time,they were divided into the better prognosis group(n=22) and the worse prognosis group(n=16),for analyzing the difference between group A and B in terms of age,preceding infections,maximal Hughes grades and neuro-electrophysiology including motor conduction velocity(MCV),distal complex muscle action potential(dCMAP) and F wave,and investigating the related factors with the prognosis of GBS.Results1.MCV of tibial nerve was(40.2?2.53) m/s and(33.4?2.46) m/s in group A and group B,respectively;MCV of peroneal nerve was(45.2?3.23) m/s and(38.3?2.16) m/s in group A and group B,respectively,and the difference between group A and group B was significant(Pa0.05);abnormal rate of F wave(68.42%) was higher than abnormal rate of MCV(42.11%) and dCMAP(42.11%)(Pa

Objective To explore the efficacy of percutaneous intratumoral injection of mixed emulsion with hot lipiodol and pira-rubicin for treatment of liver metastases lack of blood supply.Methods 12 patients with liver metastases,confirmed by the clinical and pathological examination,were treated by intratumoral injection of mixed emulsion with heating lipiodol and pirarubicin using 21G biliary needle under CT guidance.The adverse events were recorded and the treatment efficacy was also assessed according to the imaging findings.Results 8 patients achieved a partial remission of lesions,and 3 ones had stable condition.No serious adverse reactions occurred.Conclusion It is effective,simple,convenient and safe for treatment of liver metastases lack of blood supply with percutaneous intratumoral injection of emulsion with hot lipiodol and pirarubicin.