It's a multi-protein complex and not single protein that accomplish most of cellular activity.Therefore,to identify and analyze the composition of protein complex is necessary for studying the function of proteins.Tandem affinity purification technique(TAP) enables the effective purification of complex under physiological conditions without knowledge of the complex composition and function.Now,it has been adapted to analyze the composition of the protein complexes in yeast.Combined with mass spectrometry,TAP can identify the interacting proteins of target protein and offer great help for opening protein interacting network and function out.

Objective To construct an Lentiviral expression vector of pLVX‐IRES‐ZsGreen1‐MIA2 targeting to MIA2 and in‐vestigate its effect on the expression of MIA2 and growth of HCC cell line HepG2 in vitro ,observe MIA2 changes and the influence on apotheosis ,thus to provide preliminary experimental fundament for successive researching on the role of MIA2 in the pathogene‐sis of HCC .Methods The sequence of pLVX‐IRES‐ZsGreen1‐MIA2 was designed and synthesized .The pLVX‐IRES‐ZsGreen1‐MIA2 Lentiviral expression vector was constructed and then transiently transfected into HepG2 HCC cells in vitro .The proportion of pLVX‐IRES‐ZsGreen1‐MIA2 positive cells was observed under the fluorescence microscope .Then ,the expression level of MIA2 was detected by real time PCR .Moreover ,the proliferation of HepG2 cells was observed by MTT assay and colony formation as‐say .Finally ,the migration of HepG2 cells in vitro was also determined by Scratch assay .Results pLVX‐IRES‐ZsGreen1‐MIA2 Lentiviral expression vector was successfully constructed .Compared with control group (NC) ,the expression level of MIA2 was significantly decreased in transfected groups(P<0 .05);MTT assay showed that the proliferation of HepG2 cells was dramatically reduced in pIRES2‐ZsGreen1‐MIA2transfected groups(P< 0 .05);furthermore ,the number of both colony forming and migrating cells were also remarkably reduced in transfected groups(P<0 .05) .Conclusion The pIRES2‐ZsGreen1‐MIA2 can significantly re‐duce the expression level of MIA2 and inhibit the proliferation and migration of the HepG2 HCC cells in vitro .

Adult ; Aged ; Aged, 80 and over ; Cerebrovascular Circulation ; Cervical Vertebrae ; pathology ; Female ; Humans ; Male ; Manipulation, Spinal ; Middle Aged ; Spinal Osteophytosis ; diagnostic imaging ; therapy ; Tomography, Spiral Computed ; Vertebral Artery ; diagnostic imaging ; pathology ; Vertebrobasilar Insufficiency ; diagnostic imaging ; therapy

Objective To evaluate the impact of early enteral nutrition (EEN) compared with parenteral nutrition (PN) on patients after hepatectomy. Methods Seventy-eight patients undergoing liver resection were randomized prospectively into two groups: EEN group receiving early enteral nutrition (n=35) and PN group receiving parenteral nutrition (n=43). The patients in both groups received isocaloric and isonitrogenous nutritional formulas 24 h after operation and the formulas were stopped on postoperative day 7. The general conditions, liver function tests, clinical complications,and clinical nutritional variables at three time points that included preoperative phase, postoperative day 1 (POD 1) and postoperative day 8 (POD 8) were observed. Results No significant differences were found in length of hospital stay, liver function and clinical nutritional variables between the 2 groups. In the EEN group, the serum prealbumin level almost returned to the preoperative level on POD 8. The nutritional complication rate of the EEN group was increased significantly but it was milder than that of the PN group. The time of gut function recovery in the EEN group was shorter than that of the PN group. The costs of nutritional drugs showed a significant decrease in the EEN group.Conclusion Early enteral nutrition is safe, rational and effective in patients who have undergone hepatectomy. Early enteral nutrition is better than parenteral nutrition in promoting liver function recovery, liver protein synthesis, postoperative recovery of gut function and decreasing costs of nutritional drugs.

Objective To identify predictors of prescription initiation and persistence of warfarin in non-valvular atrial fibrillation ( NVAF ) patients with high risk of stroke ( CHA2 DS2-VASc≥2 ) . Methods NVAF patients consulted in our hospital from Aug , 2011 to Apr, 2015 were enrolled.Patients who underwent radiofrequency catheter ablation were excluded . Patients were divided into two groups (warfarin group and non-warfarin group).Logistic regression was used to estimate the predictors of initiation warfarin prescription.Kaplan-Meier survival and Cox proportional hazards model was performed to determine rate of warfarin persistence and its associated factors .Results A total of 622 AF patients were enrolled and 490 patients with CHA2DS2-VASc≥2.Ten patients lost follow up and 480 patients were followed up with a mean follow-up period of ( 40.0 ±11.55 ) months.Of which 245 NVAF patients ( 51%) had a warfarin prescription.Patients with ischemic stroke ( OR 2.447 , 95%CI 1.435-4.171 , P=0.001 ) , heart failure ( OR 2.009 , 95%CI 1.084-3.724 , P=0.027 ) and persistent AF ( OR 2.231 , 95%CI 1.448-3.437 , P=0.0001 ) had a higher likelihood of warfarin prescription .Anemia ( OR 0.479 , 95%CI 0.238-0.964 , P=0.039), concommitant Traditional Chinese Medicine (TCM) use (OR 0.638, 95%CI 0.456-0.891, P=0.008 ) and longer distance to hospital ( OR 0.759 , 95%CI 0.610-0.945 , P=0.014 ) decreased the likelihood of warfarin prescription . One hundred and seventy-six ( 71.8%) warfarin users continued persistent therapy and the overall proportion of warfarin persistence was 78.3% for one year , 71.0% for 3 years.Seventy-six existing warfarin users continued the warfarin therapy (80%, 76/95),one hundred new users showed persistence to therapy ( 66.7%, 100/150 ) .Warfarin use before enrollment significantly increased warfarin persistence than new prescription ( P =0.008 ) .Variables associated with higher discontinuation were new prescription ( HR 1.786 , 95% CI 1.029-3.100 , P=0.039 ) , TCM use ( HR 1.687 , 95%CI 1.201-2.37 , P=0.003 ) and longer distance to hospital ( HR 1.446 , 95% CI 1.121-1.865, P=0.005).Conclusions In anticoagulation clinic, concommitant TCM use, distance to hospital and other factors were associated with warfarin initiation prescription and persistence .Identifying factors associated with warfarin treatment could help in developing adherence of patients .

AIM: To investigate the probably mechanism of amniotic extraction inhibiting haze formation after epipolis laser in situ keratomileusis (Epi-LASIK) in rabbit cornea.METHODS: Thirty rabbit corneas were performed with Epi-LASIK.All eyes were randomly divided into three groups: eyes treated with amniotic extraction (AE group),eyes treated with 1g/L dexamethasone (hormone group) and eyes treated with solvent (solvent control group).Haze grade evaluation was performed under the slit lamp after Epi-LASIK for 1,4 and 8wk.The repair of corneal epithelium was observed by using HE staining,and the expression of NF-kB protein P65 was detected by immunohistochemistry.The expression levels of inflammatory cytokines (TNF-α,TGF-β1 and IL-1) and anti-inflammatory cytokines (IL-4,IL-10 and IL-13) were determined by ELISA.RESULTS: HE staining showed that the basal cells of corneal epithelium were more uniform and arranged regularly in AE groups after Epi-LASIK for 1wk as compared with the hormone group and the solvent control group.After 4wk,there were a few of new collagen fibers in the superficial stroma of AE group,forming a small amount of scar.After 8wk,the corneal stroma of AE group showed a small amount of new collagen fibers,arranged regularly,and rarely formed scar.At the early stage (1 and 4wk),AE treatment has an obviously effect on inhibiting the secretion of inflammatory factors (TNF-α,TGF-β1 and IL-1) and anti-inflammatory factors (IL-4,IL-10 and IL-13),and the difference was statistically significant (P<0.01).Moreover,the activation of the NF-kB signaling pathway was significantly inhibited by treatment with AE in the early postoperative period (1 and 4wk).CONCLUSION: Amniotic extraction may reduce the inflammatory response in corneal epithelial cells by inhibiting the NF-kB signaling pathway,thereby inhibiting the formation of collagen and scar and the occurrence of haze.

AIM: To explore the effect of Buddleia flavonoids drug-containing plasma and androgen receptor(AR) blocker on the expression of STAT1 phosphoprotein.METHODS: In vitro lacrimal gland epithelial cells were cultivated with H2O2 to establish the dry eye apoptosis state. Blank plasma group, Buddleia officinalis plasma total flavonoids interfere with drug-containing group, and the intervention group of testosterone propionate were set. The expressions of STAT1 phosphoprotein of each group were observed by Western blot and AR blocker flutamide was used to explore the intended androgen effect of Buddleia flavonoids.RESULTS: After the intervention of drug-containing plasma, the expression of STAT1 Phosphoprotein in Buddleja officinalis drug-containing plasma intervention group(0.353±0.494) and testosterone propionate intervention group(0.502±0.036) were enhanced and the differences between the two groups were significant (P<0.01). After using the AR blocker in all groups, the expression of STAT1 phosphoprotein in each group (0.268±0.061,0.283±0.106,0.213±0.071) had no difference.CONCLUSION: Buddleja officinalis drug-containing plasma total flavonoids can promote the expression of STAT1 phosphorylation.

Background Nd: YAG laser posterior capsulotomy is an important way for after cataract.Usually the patient will use glucocorticoid eye drops to treat the anterior chamber inflammation after operation,but there is potential risk of elevating intraocular pressure (IOP).Objective This study was to compare the clinical effectiveness and safety of loteprednol etabonate ophthalmic suspension,tobramycin+ dexamethasone eye drops and fluorometholone eye drops following Nd: YAG laser posterior capsulotomy.Methods A randomized-controlled clinical trail was performed.One hundrcd and seventy-onc cycs of 127 paticnts who received Nd: YAG laser posterior capsulotomy for after cataract were randomly divided into four groups.Loteprednol etabonate ophthalmic suspension,fluorometholone eye drops,tobramycin+dexamethasone eye drops and systane eye drops was topically administered respectively in the four groups after laser posterior capsulotomy and 6 times per day for 5 days.IOP was measured with Goldmann tomometer 1 hour before operation and 1 hour,1 day,3 days and 7 days after operation.The ocular anterior segment inflammatory response was examined under the slit lamp and scored based on the Peizeng criteria.Written informed consent was obtained from each patient before any relevant medical procedure.Results The IOP was (18.2 ±4.7),(20.1 ±5.7),(18.7±5.5),(19.0 ±4.1),(19.5 ±3.5) mmHg in various time points in the loteprednol etabonate group; (18.7 ±5.3),(20.9±5.7),(21.3±4.5),(21.0±4.9),(22.5±6.5) mmHg in the fluorometholone eye drops group ; (17.9± 6.3),(20.3 ± 6.1),(23.0 ± 3.7),(24.7 ± 4.9),(24.5 ± 6.5) mmHg in the tobramycin +dexamethasone group and(18.4±6.3),(20.7±3.7),(22.7±6.5),(19.6±4.8),(18.5±3.5) mmHg in the systane group,showing a significant difference among the 4 groups (Fgroup =3.876,P =0.023).With the time lapse,the IOP was gradually reduced in the loteprednol etabonate group and systane group,but that in the fluorometholone group and tobramycin+dexamethasone group was elevated,showing a significant difference among them (Ftime =3.801,P =0.031).No any ocular and systemic adverse effect was found in various groups.The percentage of grade 1 and 2 of aqueous inflammatory cells was lower in the loteprednol etabonate group and tobramycin+dexamethasone group than the fluorometholone group and fluorometholone group and systane group(H =8.276,P =0.012).The percentage of Ⅰgrade of aqueous flare was 8％ in the loteprednol etabonate group,22％ in the fluorometholone group,18％ in the tobramycin+dexamethasone group and 30％ in the systane group,with a significant difference among them (H=9.305,P=0.000).Conclusions The use of corticosteroid eye drops can relieve the inflammatory response of ocular anterior chamber after Nd: YAG laser posterior capsulotomy.Loteprednol etabonate ophthalmic suspension has a better anti-inflammatory effect and less influence on IOP.

Objective:To analyze the relationship between hepatitis B virus(HBV)gene mutation at 1896 in precore region with genotype and replication of HBV and the liver function of patients.Methods:HBV precore 1896 site mutation,the genotype of HBV and serum content of HBV DNA were determined by PCR in 60 patients positive of HBV DNA.Chemiluminescence miacropaticle immunoassay(CMIA)was used for detection of serum HBeAg and HBeAb.Liver function parameters were ob- tained by routine biochemistry method.Results:The alanine aminotransferase(ALT)level in HBV with 1896 site mutation was significantly higher than that in the wildtype virus.Site mutation at 1896 had no correlation with HBeAg,HBV genotype and HBV DNA content.HBV DNA content in patient with genotype C was significantly higher than that with genotype B(P

Anastomosis, Surgical ; Anastomotic Leak/surgery* ; Drainage ; Gastrectomy ; Humans ; Suction