MTM1 gene is essential for SOD2 activity and normal mitochondrial function. MTM1 deletion results in decreased SOD2 activity, impaired mitochondrial function and growth defect on nonfermentable carbon source. A yeast genomic library was transformed into mtm1 deletion mutant to screen for suppressor genes of MTM1. The damage caused by MTM1 deletion is irreversible and even overexpression of MTM1 can not rescue the growth defect of mtm1 deletion mutant. Another screening strategy was adopted: a plasmid overexpressing MTM1 was transformed into wild type before the MTM1 gene on chromosome was deleted. The resulting strain, designated YES2MTM1, was transformed with a yeast genomic library. Transformants lost the plasmid overexpressing MTM1 after 5-FOA treatment. Yeast strains able to grow on nonfermentable carbon source with MTM1 deletion and overexpression of some DNA fragments were picked up and candidate suppressor genes were identified. Overexpression of five genes were identified to be able to rescue the growth defect on nonfermentable carbon source. The study will provide reference for MTM1 gene function and screening for suppressor of genes whose deletion result in irreversible damage.

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 develop an indirect enzyme-linked immunosorbent assay (ELISA) for semi-quantification of major allergen parvalbumin in fish.Methods:The soluble proteins were prepared from both white and dark muscles of seven species of freshwater fish and five species of marine fish.Tricine-SDS-PAGE and Western blot were performed to examine the protein patterns of fish muscle extracts.Natural parvalbumin being used to make calibration curve was purified from silver carp (Hypophthalmichthy molitrix) by ammonium sulphate fractionation,followed by ion exchange and gel filtration chromatography.The molecular mass of purified protein was estimated by Tricine-SDS-PAGE and identified by Western blot with anti-frog parvalbumin monoclonal antibody PARV-19.ELISA using PARV-19 was carried out to evaluate parvalbumin contents in white and dark muscles.Results:Tricine-SDS-PAGE revealed species-specific differences in proteins of heated extracts.Western blot confirmed that the major bands were showed in Tricine-SDS-PAGE with the molecular masses of 10-14 kD corresponded to parvalbumins recognized by PARV-19 and various numbers of isoforms of parvalbumin existed in different species of fish.There might be some differences in the parvalbumin contents and the epitope region was recognized by PARV-19 based on the differences in relative intensities of protein immunodetection.The ELISA showed that the contents of parvalbumin were 4 to 33 folds higher in the white muscle than in the dark muscle and varied greatly in different species of fish.Conclusion:These results validate that the dark muscle might be less allergenic than the white muscle due to the lower content of parvalbumins,and it is suggested that the commercial anti-parvalbumin antibody PARV-19 can be used to detect parvalbumins from the commercially important species of fish tested in this study and the method we develope succeeds to detect the major allergen in various species of fish.

Objective To evaluate the effect and mechanism of bone morrow MSCs transplantation in swine with AMI by cell biology and molecular imaging methods including PET/CT, SPECT, and MRI. Methods Twenty?four Chinese mini?swine ( ( 25 ± 5 ) kg ) were randomly divided into 2 groups: MSCs group ( n=12) and control group ( n=12) . Myocardial infarction was induced in swine hearts by occlusion of the LAD. Thirty minutes later, the MSCs group received autologous MSCs transplantation through in?tramyocardial injection into the peri?infarcted areas (2×107,2 ml) and the control group was subjected to cell culture medium in the same way. At the 1st and 4th weeks after MSCs transplantation, myocardial glu?cose metabolism, myocardial perfusion and cardiac function were evaluated in the two groups through PET/CT, SPECT and MRI. The minimum FDG mean signal intensity ( MSI ) , summed MSI, SRS, SRS%, LVEF, ESV, stroke volume ( SV) and cardiac output ( CO) were calculated. On the 4th week, HE and Masson′s Trichrome stains were performed. Mann?Whitney u test and non?parametric Wilcoxon test were used. Results (1) As evaluated by PET in the 1st week, the MSI and summed MSI in MSCs group were less than those in control group ( 22. 10 ± 3. 18 vs 35. 70 ± 3. 02, z=-2. 65; 1 013. 50 ± 29. 37 vs 1 084. 00 ± 21?15, z=-1.97;both P<0.05) . Compared to the minimum MSI and summed MSI in the 1st week, those in MSCs group increased significantly (34.00±4.25, z=-2.81;1 075.50±28.30, z=-2.80;both P<0?01) in the 4th week. SRS and SRS% decreased in the 4th week compared to those in the 1st week (20.20±2.24 vs 23.80±1.58, (29.80±3.31)% vs (35.10±2.34)%;both z=-2.08, both P<0.05). The averaged MSI in left ventricular infarction area (MSI<70) also increased (56.25±3.54 vs 48.14±2.71;z=-2.80, P<0.01). The a?bove?mentioned parameters had no statistically significant differences in the 4th week compared to those in the 1st week in the control group (all P>0.05). (2) In the 1st week, the perfusion variables had no signifi?cant differences between the two groups ( P>0.05) . There was no significant difference in any perfusion vari?ables between the 1st and 4th weeks in the two groups, respectively (P>0.05). (3) As evaluated by MRI, the cardiac functional parameters had no significant differences between the two groups at the 1st week. In the MSCs groups, LVEF increased significantly ((54.41±2.62)% vs (47.54±2.43)%;z=-2.60, P<0.01) and ESV reduced significantly ((22.85±1.91) vs (27.07±1.67) ml;z=-2.70, P<0.01) in the 4th week com?pared to those in the 1st week; SV and cardiac CO in the 4th week also increased significantly ((29.35± 1?84) vs (26.52±1.46) ml, (2.23±0.14) vs (1.96±0.13) L/min;z=-2.09 and -1.99, both P<0?05). In the control group, there were no significant differences in the cardiac functional parameters between the 1st and 4th weeks ( all P>0.05) . Conclusions Four weeks after MSCs transplantation for AMI, cardiac func?tion and myocardial glucose metabolism improved significantly but without significant myocardial perfusion improvement. Therefore, the cardiac function improvement might be associated with increased myocardial glucose metabolism.

Objective To explore the effects of regular doses of proton pump inhibitors (PPIs) on intestinal barrier function in the elderly.Methods The elderly hospitalized patients with acidrelated disorders were retrospectively analyzed from April to December 2016 in Department of Geriatrics,the General Hospital of Guangzhou Military Command.Based on the criteria of inclusion and exclusion,all patients received the stomachoscopy examination,and received PPI orally for more than 1 month.The treatment continued for 1-6 months.The blood status was routinely examined at pre-and post-treatment,including the level of liver function,C reactive protein (CPR),procalcitonin (PCT),diamine oxidase (DAO),D-lactate acid and serum endotoxin.Results A total of 88 elderly patients aged 80-96 years were involved.Most patients took PPI for 1 month.The levels of alanine aminotransferase,aspartate amino transferase,CPR and PCT were mostly normal after 1 month of treatment.Meanwhile,there were the comparabilities between the post-and pre-treatment groups in levels of DAO[(6.53±3.00) vs.(6.23±2.42) U/L;t=-0.91,P=0.37],of D-lactate acid[(7.31 ±3.67) vs.(7.62±3.35) mg/L;t=0.73,P=0.47]andof serum endotoxin[(11.69±5.98) vs.(10.97±5.76) U/L;t=-1.66,P=0.10]without statistical significance.Additionally,after 23 patients took PPI for 6 months,there were no statistical significances between the pre-and post-treatment groups in the levels of DAO[(5.35±2.49) vs.(6.59±2.69)U/L,t=1.89,P=0.07],of D-lactate acid[(8.12±3.84) vs.(7.71±3.65) mg/L,t=-0.73,P=0.47]and of serum endotoxin[(10.23 ± 5.51) vs.(10.41±6.45) U/L,t=0.18,P=0.86].Conclusions There is no significant effect on intestinal permeability and intestinal mucosa barrier function in the elderly taking PPI for 6 months.New studies are needed to understand the role of PPI in the intestinal microflora in the elderly.

Objective To study the surgical treatment of xanthogranulomatous cholecystitis (XGC).Methods We retrospectively analyzed the clinical data of 56 patients with XGC who underwent surgical treatment at the Zhejiang Provincial People's Hospital from May 2010 to May 2017.Results The diagnosis of XGC was confirmed by histopathology.On preoperative examination of the 56 patients,42 patients had various degrees of increase in the CA19.9 levels,41 patients (73.2％) had thickened gallbladder walls with continuous mucosal linings on ultrasonography,CT,or MRI,and 18 patients (32.1％) had thickening of gallbladder walls with low density nodules.Gallbladder stones were present in 51 patients (91.1％) and 4 patients (7.2％) presented with Mirizzi syndrome.The 41 patients (73.2％) who were diagnosed as XGC before operation under laparoscopic surgery and 7 patients (17.1％) were converted to open surgery.The remaining 15 patients (26.8％) underwent open operation directly because of uncertainty in the diagnosis.All the patients had frozen section during operation.The postoperative pathological results included 21 localizedtype (37.5％) and 35 diffuse type (62.5％) of XGC.All 56 patients had no long-term complications on followed-up for 0.5～ 6 years.Conclusions XGC is a special kind of chronic cholecystitis.There is difficulty in differentiating from gallbladder cancer before surgery.The diagnosis of XGC mainly depends on ultrasonography,CT or MRI.Cholecystectomy is the treatment for XGC.Laparoscopic surgery is the first line treatment for XGC.

Objective To explore the safety and feasibility of laparoscopic hepatectomy combined with radiofrequency ablation for primary liver cancer difficult to manage.Methods A retrospective study was conducted to analyze the clinical data of 16 patients who underwent laparoscopic hepatectomy combined with radiofrequency ablation in the Zhejiang Provincial People's Hospital from Apr 2015 to Dec 2017.Results 2 more tumors were found by intraoperative laparoscopic ultrasound.All patients underwent laparoscopic hepatectomy combined with radiofrequency ablation successfully.There were 2 laparoscopic left hepatectomy combined with radiofrequency ablation,2 laparoscopic right hepatectomy combined with radiofrequency ablation,6 laparoscopic left lateral lobectomy combined with radiofrequency ablation,4 laparoscopic right postrior lobectomy combined with radiofrequency ablation and 2 irregular laparoscopic hepatectomy combined with radiofrequency ablation.The intraoperative blood loss ranged from 100-800 ml.The average operative time was (283 ± 112) min.The length of postoperative hospital stay ranged from 5 to 12 days.The tumor-free survival rate after operation is 100％ (16/16) on 6 to 38 months follow up.Conclusion It is safe and feasible to carry out laparoscopic hepatectomy combined with radiofrequency ablation in difficult to manage primary liver cancer.

Objective To study the safety and feasibility of laparoscopy combined with holmium laser in the treatment of chronic pancreatitis complicated with pancreatic ductal stones.Methods To compare the clinical data in patients who underwent laparoscopy combined with holmium laser (10 patients,group A) with those who underwent laparoscopy only (21 patients,group B) at Zhejiang Provincial People' s Hospital from January 2012 to August 2018.The operation time,intraoperative blood loss,intraoperative conversion rate,pancreatic ductal incision length,postoperative pancreatic fistula rate,length of postoperative hospital stay,residual stone rate and relief of postoperative abdominal pain rate of the two groups were documented and analyzed.Results Three of 31 patients were converted to open surgery.The remaining patients in the two groups were discharged home without any perioperative death.Group A and B were significant differences in the pancreatic ductal incision length (5.0±0.8 vs.6.5±1.0) cm,operation time (289.3±51.6 vs.349.5± 34.7) min,and postoperative hospital stay (8.0± 1.2 vs.10.2± 1.6) d between the two groups (P＜0.05).There were no significant differences in the intraoperative conversion to open rate,intraoperative blood loss,postoperative pancreatic fistula rate,residual stone rate and relief of postoperative abdominal pain rate between the two groups (P ＞ 0.05).Conclusions It was safe and feasible to treat chronic pancreatitis complicated with pancreatolithiasis by laparoscopy.Laparoscopy combined with holmium laser had the added advantages of easy access through the pancreaticojejunostomy,shorter operation time,and less intraoperative blood loss.

Objective:To analyze the prognosis-related factors of patients with surgical treatment of lung cancer brain metastases.Methods:From January 2016 to November 2018, the clinical data of the patients with lung cancer brain metastases received surgical treatment in Department of Neurosurgery, Cancer Hospital of Chinese Academy of Medical Sciences and Peking Union Medical College were retrospectively collected, and completed data of 83 patients were obtained. The single factor analysis was based on the log-rank method, and the multi-factor analysis was used by the Cox proportional hazard model, and the R software was used to map the influencing factors into the nomogram and verify them.Results:The median overall survival (OS) of 83 patients was 27.0 months. The median OS of patients with surgery alone was 15.7 months, the median OS of patients with surgery combined with radiotherapy, chemotherapy or targeted therapy was 27.7 months, and the difference was statistically significant ( χ2=8.735, P=0.003). The results of single factor analysis showed that gender ( χ2=4.652, P=0.031), smoking history ( χ2=8.239, P=0.004), postoperative targeted treatment ( χ2=13.697, P<0.001), postoperative adjuvant therapy ( χ2=8.735, P=0.003), pathology of metastatic tumor ( χ2=11.799, P=0.001), and lung cancer molecular graded prognostic assessment (Lung-molGPA) sore ( χ2=11.333, P=0.004) affected patients′ OS. The results of multivariate analysis showed that smoking history ( HR=0.311, 95% CI: 0.107-0.901, P=0.031), postoperative targeted therapy ( HR=3.563, 95% CI: 1.286-9.868, P=0.015), pathology of metastatic tumor ( HR=0.364, 95% CI: 0.137-0.965, P=0.042), Lung-molGPA score ( HR=0.595, 95% CI: 0.374-0.946, P=0.028) were independent prognostic factors for OS of patients with lung cancer brain metastases. In order to further evaluate the prognosis of patients, nomogram was drawn using these four independent prognostic factors. The model had high accuracy and could better evaluate the prognosis of patients. Conclusion:Lung cancer brain metastases patients with operative indication can benefit from surgery, and active adjuvant therapy after operation can further prolong the OS of patients. The nomogram constructed by smoking, targeted therapy, pathology of metastatic tumor and Lung-molGPA score can be used to evaluate individual patient outcomes and guide clinical treatment.