Objective: To study the effect of docetaxet (DOC) combined with 4-AP on human breast can-cer MCF-7 cells and to explore whether 4-AP could strengthen the effect of docetaxel. Methods: MTT assays were performed to investigate the effect of docetaxel, 4-AP and the combination of them on the proliferation of MCF-7 cells. Flow cytometry was employed to detect cell cycles and cell apoptosis after the cells were stained by PI alone or by Annexin-V and PI. Results: Docetaxel could significantly inhibit the proliferation of MCF-7 cells in a dose- and time- dependent manner. 4-AP could inhibit the proliferation of MCF-7 cells and the inhibitory rates were 11.9%±1.7%, 42.1%±3.2%, and 44.2%±1.6% at 24h, 48h and 72h after adding 4-AP. Moreover 4-AP (5mmol/L) could strengthen the effect of docetaxel. 4-AP (25μmol/L) could increase the effect of Docetaxel. Docetaxel at 5μmol/L could significantly increase the percentage of cells at G_2/M (53.58%± 1.44% vs. 8.83%±0.44%, P<0.01) and decrease the percentage of cells at G_0/G_1 (11.48%±0.14% vs. 63.89%±0.98%, P<0.01), indicating that docetaxel blocked MCF-7 cells at G_2/M phase. 4-AP at 5mmol/L could in-crease the percentage of MCF-7 cells at G_0/G_1 and decrease the percentage of cells at G_2/M (0.42%±0.17% vs. 8.83%±0.44%, P<0.05). Docetaxel could significantly increase late apoptosis and death of MCF-7 cells af-ter treatment over 24h (from 6.97%±0.75% to 20.77%±0.75%, P<0.05). Docetaxel combined with 4-AP could increase early apoptosis rate from 4.60%±0.91% to 12.20%±0.82% (P<0.05) and could increase late apopto-sis rate and death rate from 4.60%±0.91% to 12.20%±0.82% (P<0.05). Conclusion: Both docetaxel and 4-AP can inhibit the proliferation of MCF-7 cells. Docetaxel can increase the percentage of cells at G_2/M phase and 4-AP can increase the percentage of cells at G_0/G_1 phase. 4-AP could strengthen the inhibitory effect of docetaxel on the proliferation of MCF-7 cells through inducing cell apoptosis.

A stimulate method for determination of polybrominated diphenyl ethers ( PBDEs) and derivatives (OH-PBDEs and MeO-PBDEs), tetrabromobisphenol A (TBBPA), hexabromocyclododecane (HBCD) in egg samples was developed by gel permeation chromatography ( GPC) and dispersive solid phase extraction ( DSPE) combined with liquid chromatography tandem mass spectrometric ( HPLC-MS/MS) and gas chroma-tography-negative chemical ionization mass spectrometry ( GC-NCI/MS ) . The analytes were extracted with mixture of hexane and dichloromethane (1∶1, V/V) by accelerated solvent extraction (ASE), and purified by 100 mg C18 dispersive solid phase extraction ( SPE) sorbents followed with gel permeation chromatography (GPC) , and then analyzed by liquid chromatography tandem mass spectrometric (HPLC-MS/MS) and gas chromatography-negative chemical ionization mass spectrometry (GC-NCI/MS), respectively. The quantita-tion was carried out external standard method. The recoveries of objects were 64. 5%-97. 2% and 65. 6%-109 . 2% ( except BDE85 was 54 . 8%, OH-BDE-137 was 47 . 4%) spiked at 1 . 0 μg/kg or 5 . 0 μg/kg in egg white and egg yolk, respectively. The relative standard deviations (RSDs) were less than 20. 2%. The limits of quantitation (LOQ) for the object were 0. 01-0. 2 μg/kg.

Objective To define the assessment dimensions of community-based elderly care service.Methods Firstly 11 assessment dimensions of elderly care service needs were teased out through literature review and group discussion, these were illness, self-care ability, vision, risk of falls, willingness/information self-expression ability, the comprehension in others' willingness/information, depression, anxiety, family care ability, self-desires for old-age support, family-desires for old-age support.Then 37 experts were invited to reconfirm and empower theose assessment dimensions with two rounds of Delphi method.Results Results showed that the enthusiasm and authority degree of experts were favorable in two rounds, and the results from the second round showed strong opinion coordination.Finally, 12 assessment dimensions were confirmed, in which financial situation of the aged added and used for reference.After weight calculation, illness was weighted the highest (0.11), while vision and the degree of anxiety were weighted lowest in 11 dimensions (0.07).Conclusion The reliability of the assessment dimensions of the aged service needs is high.This study had positive significance on the solution of the dilemma in elderly care at the present stage.

Objective To investigate whether metabolic pathway-related gene polymorphisms are associated with arterial plaque stability and their gene-gene interactions increase the risk of cerebral infarctions.Methods Totally 294 patients with atherothrombosis stroke admitted to the Department of Neurology, the Third Affiliated Hospital of Wenzhou Medical University from September 2010 to December 2012 were divided into a carotid vulnerable plaque group ( n=69 ) and a stable plaque group ( n=225 ) according to the results of carotid B-mode ultrasonography.A total of 282 healthy volunteers excluded carotid plaque and stroke were enrolled as well.Genetic polymorphisms of ALOX5AP and CYP3A5, CYP2C9*2, CYP2C9*3 and EPHX2 were genotyped using polymerase chain reaction and mass spectrometry analysis.The SPSS16.0 software was used to compare genotype frequencies and the generalized multifactor dimensionality reduction ( GMDR ) method was applied for gene-gene interaction analyses.Results The results showed that EPHX2 GG genotype might protect against stroke ( OR =0.520, 95% CI 0.288 -0.940, P=0.030).The distribution of CYP3A5 genotypes showed statistically significant differences (χ2 =7.284, P=0.026) between the vulnerable plaque ( AA: 5 cases, AG: 36 cases, GG: 28 cases) and stable plaque ( AA: 26 cases, AG: 77 cases, GG: 122 cases ) groups.Multivariate Logistic regression analysis showed that the GG genotype of CYP3A5 was protective factor for unstable plaques ( OR=0.405, 95%CI 0.178 -0.920, P =0.031 ).Differences in other SNPs did not reach statistical significance between the two groups.The GMDR analysis showed a significant gene-gene interaction between SG13S114 and A6986G, with scores of 10 for cross-validation consistency and 9 for the sign test (P=0.011).The best model for ischemic stroke was found to be SG13S114 AA and A6986G AA.Adjusting for age, hypertension and diabetes, the certain gene-gene interaction predicted a significantly higher risk of cerebral infarction (OR=1.804, 95%CI 1.180-2.759, P=0.006).Conclusions The EPHX2 G860A gene might be linked with the incidence of cerebral infarctions.Only a CYP3A5 gene polymorphism might be associated with carotid plaque instability in patients with stroke.The gene-gene interaction predicts a significantly higher risk of cerebral infarction.There is a 1.804-fold increased risk for ischemic stroke in individuals with these combined genetic factors.

Objective To evaluate the result of multiple segmentectomy under selective occlusion of hepatic inflow for complicated intrahepatic cholangiolithiasis. Methods In this study 12 cases of complicated intrahepatic cholangiolithiasis receiving multiple segmentectomy under selective occlusion of hepatic inflow during 2004. 1 - 2010. 10 were reviewed retrospectively. The short-term and long-term outcomes of the patients were analyzed. Results There was no surgical mortality in this group. The segmentectomy performed were Ⅱ + Ⅲ + Ⅴ in 1 case; Ⅱ + Ⅲ + Ⅵ in 2 cases; Ⅱ + Ⅲ + Ⅴ + Ⅵ in 1 case;Ⅱ +Ⅲ+Ⅵ +Ⅶ in4 cases; Ⅱ +Ⅲ +Ⅳ +Ⅵ in 3 cases and Ⅱ + Ⅲ +Ⅳ +Ⅵ +Ⅶ in 1 case. The average intraoperative blood loss was 560 ± 291 ml. Postoperative complications were wound infection in 2 cases, bile leakage in 1 case, abdominal infection in 1 case. There were no liver failure, intrabdominal hemorrhage or hemobilia; stone clearance rate at 10 days after operation was 83% (10/12) and 92%(11/12) at 6 weeks after operation following postoperative choledochoscopic lithotripsy. 92% (11/12) cases were followed-up with the median follow-up period of 31 months. The result was excellent or good in 92% (11/12) cases. Conclusions Multiple segmentectomy was the choice for complicated intrahepatic cholangiolithasis, and the procedure could be safely performed under selective occlusion of the hepatic inflow.

Objective To establish cell models with different insulin sensitivity status and examine the expression of protein kinase B (PKB/Akt) in the same conditions. Methods 3T3-L1cells were cultured with different glucose concentrations (3.0, 5.5 and 50.0 mmol/L respectively).Insulin sensitivity, as presented by insulin-induced glucose transport rate in the cells, was tested by 3H-2-DG incorporation. PKB expression in cells was determined by RT-PCR. Results The insulin-induced glucose transport rate in 3T3-L1 cells was affected by different concentrations of glucose. The transport rate of 3H -2-DG in cells cultured in low glucose concentration was increased by 152% and 50% compared with high glucose concentration and control group, respectively. There was no difference in PKB mRNA expression among the three groups under the same conditions,whereas there was an obvious reduction of PKB expression in the three groups after the cells were treated with glucose and insulin. Conclusions 3T3-L1 cells cultured with different concentrations of glucose present the different insulin sensitivity status, but the difference of PKB mRNA expression has not been observed in the same condition.

Objective: To evaluate the changes of left and right ventricular function in patients after percutaneous pulmonary valve implantation (PPVI). Methods: A total of 8 patients with successful PPVI in our hospital from 2014-05-27 to 2015-03-25 were studied. The patients received pre-operative evaluation including clinical symptoms, plasma BNP levels, ECG, chest X-ray, echocardiography, CT and MRI examinations. Post-operative follow-up study was conducted at 6 months after PPVI to compare the ventricular functional changes. Results: The average age in 8 patients was 14-42 (25.4±8.1) years including 7 with tetralogy of Fallot correction and 1 with pulmonary stenosis plasty for 24 years. Echocardiography found that 3 patients with mid to large pulmonary regurgitation and 5 with large regurgitation. Compared with pre-operation, at 6 months after PPIV, all patients had decreased right ventricular end-diastolic diameter (RVEDD) as (44.0±4.8) mm vs (33.6±7.1) mm, right ventricular end-diastolic volume index (RVEDVI) (150.1±25.7) ml/m2 vs (111.4±39.1) ml/m2, RVESVI (107.8±21.5) ml/m2 vs (80.7±22.2) ml/m2 and right ventricular cardiac output (RV-CO) (6.8±1.3) L/min vs (4.9±0.8) L/min, but right ventricular ejection fraction (RVEF) was similar (40.5±6.2) % vs (39.5±9.9) %, P>0.05; while
increased LVEDD (42.9±4.4) mm vs (46.1±3.0) mm, P<0.05, but LVEDVI (61.8±15.0) ml/m2 vs (72.4±17.6) ml/m2, LVESVI (47.8±12.4) ml/m2 vs (41.0±10.4) ml/m2, LVEF (50.9±5.5) % vs (52.8±6.7) % and LV-CO (3.7±1.0) L/min vs (4.2±1.0) L/min were similar, allP>0.05. Conclusion: PPVI may decrease right ventricular preload, improve its reverse remodeling and maintain systolic function at normal level in relevant patients; while the impacts on left ventricular function and geometry should be further studied.

Objective To investigate the ability of different standards on the evaluation of vascular invasion of pancreatic carcinoma. Methods The MSCT imagings of 56 cases of pancreatic carcinoma confirmed by surgery were analyzed retrospectively. The vascular invasion degrees of 5 large vessels around the pancreas were evaluated according to Loyer classification standard, Lu classification standard and the Changhai standard, the surgical result was used as the gold standard. The accuracy of different standards was evaluated, and the Kappa coefficient were calculated. Results The accuracy, sensitivity, specificity, positive predictive value and negative predictive value of Loyer classification standard were 86.79%, 86.27%, 86.90%, 59.46% and 96.60%, respectively, and the Kappa coefficient was 0.623. The accuracy, sensitivity, specificity, positive predictive value and negative predictive value of Lu classification standard were 93.21%, 84.31%, 95.20%, 79.63% and 96.46%, respectively, and the Kappa coefficient was 0.777. The accuracy, sensitivity, specificity, positive predictive value and negative predictive value of Changhai standard were 95.36%, 84.31%, 97.82%, 89.58% and 96.55%, respectively. The Kappa coefficient was 0.841. Conclusions The Changhai standard was feasible to evaluate the vascular invasion of pancreatic carcinoma.

Objective To evaluate the efficacy of MRI for assessment of re-distribution of bone marrow mesenchymal stem cells injected intramyocardially in main organs (heart, liver, spleen and kidney) under different heart status (beating or arresting) in a porcine model. Methods Bone marrow-derived mesenchymal stem cells were obtained from the male swine and labeled with iron oxide during culture. Acute myocardial infarction was created in female swine, one week later, the survivors were randomly divided into 4 groups. Cardiopulmonary bypass was set up to arrest the heart, and then labeled cells (1×108) were intramyocardially injected into the border of the infracted myocardium in group 1 (n=6). The same volume of cells was grafted into the beating heart in group 2 (n=6). In group 3 and 4, saline was injected into either the arresting or beating myocardium. Three days later, re-distribution of stem cells and cardiac function were assessed by T2*WI and cine MRI, respectively. All animals were sacrificed for histology and real-time quantitative polymerase chain reaction (RT-PCR) of sex-determining region on Y-chromosome (SRY) investigation.The ANOVA and t test was used for statistics. Results The left ventricular end-diastolic volume (LVEDV) before transplantation for group 1-4 were: (56.8±5.3),(54.8±6.8),(57.4±4.3)and(56.8±2.8) ml, and after transplantation for group 1-4 were: (65.2±5.2),(63.2±3.7),(60.2±4.7)and(62.2±4.4) ml. The left ventricular end-systolic volume (LVESV) before transplantation for group 1-4 were: (33.5±7.6),(32.3±5.3),(33.5±3.6)and(32.7±4.6) ml,and after transplantation for group 1-4 were: (37.3±5.6),(36.3±6.9),(34.3±5.4)and(36.3±8.1) ml. The left ventricular EF values (LVEF) before transplantation for group 1-4 were: (42.3±7.2)%,(41.7±6.8)%,(41.8±8.6)% and(42.7±7.7)%,and after transplantation for group 1-4 were: (44.5±8.7)%,(43.1±7.4)%,(42.8±5.6)% and(43.3±8.4)%. The myocardial infarction area (MI) before transplantation for group 1-4 were: (6.5±2.1),(6.4±1.9),(6.5±2.5)and(6.4±2.6) cm2,and after transplantation for group 1-4 were: (6.4±2.3),(6.2±2.6),(6.3±2.5)and(6.4±2.8) cm2 . There were no statistical differences before and after transplantation in these 4 groups[P values of before and after transplantation for LVEDV, LVESV, LVEF,MI were >0.05 (F= 0.277, 0.066,0.066, 0.003); and >0.05 (F= 1.137,0.182,0.021,0.008),respectively]. The T2 value of the infracted myocardium in group 1 decreased more obviously than that in group 2[(-22.3 ± 2.2) vs (-17.0 ± 0.8) ms, t=-5.489, P＜0.01], while the T2 value of the spleen decreased more significantly in group 2 than that in group 1[(-7.7 ± 0.7) vs (-13.3 ± 1.1) ms,t=9.055, P＜0.01]. The T2 values of the liver and kidney were no significant differences in group 1 and 2 (liver, t=-0.532,P>0.05 and kidney, t=-0.113,P>0.05). The results of RT-PCR in group 1 and 2 showed significant differences in heart[(150±62) vs (72±4) U/L ,P<0.05, t=3.109], spleen[(131±1) vs (233±17) U/L, P＜0.01, t=- 13.286]and liver[(17±1) vs (9±5) U/L ,P<0.01,t= 3.492]. Pathological examination demonstrated that the transplanted stem cells were positive for Prussian blue staining, which had a good correlation with MRI results. Conclusion MRI can serve as a convenient and efficient imaging method to track the migration of stem cells with SPIO labeled in early stage and evaluate its early re-distribution in vivo. Injection of bone marrow mesenchymal stem cells in the arresting heart could favor retaining more cells in the myocardium.

Objective:To investigate the relationship between enteral nutrition-related diarrhea and intestinal flora in critically ill patients and the effect of microflora transplantation.Methods:A total of 60 critically ill patients with enteral nutrition-related diarrhea who were scheduled to undergo microflora transplantation in Taizhou Hospital of Integrated Traditional Chinese and Western Medicine from January 2020 to August 2021 were prospectively and continuously selected as the research group, and 60 critically ill patients without enteral nutrition-related diarrhea were selected as the control group. The bacterial count of 4 kinds of intestinal flora in the feces including bifidobacterium, lactobacillus, enterococcus, and escherichia coli were detected and compared between the two groups, and the value of the fecal colony numbers of 4 kinds of intestinal flora in diagnosing non-enteral nutrition-related diarrhea in critically ill patients was analyzed by receiver operating characteristic (ROC) curve. All patients in the research group received microflora transplantation, and the diarrhea score, hematochezia score, partial Mayo score and European five-dimension health scale (EQ-5D) were detected and compared before treatment, 1 week after treatment and 1 month after treatment to evaluate the treatment effect. The Pearson linear correlation method was used to analyze the relationship between the colony count of 4 kinds of intestinal flora colonies in the feces of the research group at baseline and the therapeutic indexes for 1 week and 1 month after treatment.Results:The number of fecal bifidobacterium and lactobacillus colonies in the study group were lower than those in the control group: (7.12 ± 0.58) × 10 7 cfu/L vs. (11.85 ± 1.25) × 10 7 cfu/L, (8.78 ± 1.05) × 10 7 cfu/L vs. (11.25 ± 1.57) ×10 7 cfu/L. The colony number of enterococcus and Escherichia coli were higher than those of control group: (8.58 ± 0.88) × 10 7 cfu/L vs. (3.84 ± 0.72) ×10 7 cfu/L, (8.25 ± 0.97) ×10 7 cfu/L vs. (3.66 ± 0.63) ×10 7 cfu/L. The differences were statistically significant ( P<0.05). ROC curve analysis results showed that the area under the curve of fecal bifidobacterium, lactobacillus, enterococcus and escherichia coli colonies in diagnosing patients with enteral nutrition-related diarrhea were all >0.7, which had certain diagnostic value. The diarrhea scores, stool blood scores and some Mayo scores of the study group at 1 week and 1 month after treatment were lower than those before treatment: (1.52 ± 0.36) and (1.13 ± 0.24) points vs. (2.45 ± 0.51) points, (0.95 ± 0.28) and (0.77 ± 0.21) points vs. (2.39 ± 0.54) points, (4.17 ± 1.24) and (3.26 ± 0.85) points vs. (7.86 ± 1.82) points, and the EQ-5D score of patients 1 week and 1 month after treatment was higher than that before treatment: (0.66 ± 0.11) and (0.79 ± 0.13) points vs. (0.58 ± 0.08) points, the difference was statistically significant ( P<0.05). Conclusions:The intestinal flora of critically ill patients is closely related to enteral nutrition-related diarrhea, and can affect the therapeutic effect of bacterial flora transplantation and the health status of patients.