The phenylacetone monooxygenase, isolated from Thermobifida fusca, mainly catalyzes Baeyer-Villiger oxidation reaction towards aromatic compounds. Met446 plays a vital role in catalytic promiscuity, based on the structure and function of phenylacetone monooxygenase. Mutation in Met446 locus can offer enzyme new catalytic feature to activate C-H bond, oxidizing indole to finally generate indigo and indirubin, but the yield was only 1.89 mg/L. In order to further improve the biosynthesis efficiency of the whole-cell catalyst, metabolic engineering was applied to change glucose metabolism pathway of Escherichia coli. Blocking glucose isomerase gene pgi led to pentose phosphate pathway instead of the glycolytic pathway to become the major metabolic pathways of glucose, which provided more cofactor NADPH needed in enzymatic oxidation of indole. Engineering the host E. coli led to synthesis of indigo and indirubin efficiency further increased to 25 mg/L. Combination of protein and metabolic engineering to design efficient whole-cell catalysts not only improves the synthesis of indigo and indirubin, but also provides a novel strategy for whole-cell catalyst development.
Escherichia coli ; genetics ; metabolism ; Glucose ; metabolism ; Indigo Carmine ; metabolism ; Indoles ; metabolism ; Industrial Microbiology ; methods ; Metabolic Engineering ; Metabolic Networks and Pathways ; Protein Engineering

Objective To investigate the bacterial spectrum and antimicrobial resistance of pathogens isolated from stool of acute diarrhea outpatients ,and provide scientific evidence for clinic rational use of antibiotics .Methods Bacteria was detected by conven‐tional feces culture method ,including separation and biochemistry appraisal sure strains .The predominant bacteria were conducted antimicrobial resistance testing in acute diarrhea outpatients .Results 544 stool specimens were collected from acute diarrhea outpa‐tients from January 2011 to December 2012 .The total positive rate was 17 .83% .Positive rates of Escherichia coli ,Salmonella , Campylobacter ,Vibrio parahaemolyticus ,Other Aeromonas ,Shiga Plesiomonas ,Shigella and Aeromonas hydrophila were 4 .78% ,3 .68% ,2 .57% and 2 .39% ,1 .84% ,1 .28% ,0 .92% and 0 .37% ,respectively .Salmonella ,Campylobacter and Vibrio parahaemolyticus were susceptible to Ofloxacin ,Amoxicillin ,Ceftazidime .They were different resistance to conventional antibiot‐ics ,which were commonly used by clinic ,and the most serious resistance are ampicillin and nalidixic acid .Conclusion Escherichia coli ,Salmonella ,Campylobacter and Vibrio parahaemolyticus are predominant bacteria pathogens .It is important to better under‐stand pathogens spectrum and antimicrobial resistance of bacteria for controlling infection in acute diarrhea outpatients .

Objective To discuss the nursing care for patients with advanced hepatocellular carcinoma (HCC) who are receiving transcatheter arterial chemoembolization (TACE) combined with sorafenib. Methods A total of 23 consecutive patients with advanced HCC who met the inclusion criteria were enrolled in this study. TACE was carried out in all patients. Three-five days after TACE the patients started to orally take sorafenib. During the treatment course, the patients were kept under close observation for adverse reactions and complications. After leaving the hospital the patients were followed up by the extended care team members, and health education as well as appropriate nursing intervention was carried out. Results All patients with advanced HCC took sorafenib orally after TACE. The major adverse events and complications were gastrointestinal adverse reactions (n = 22) and transient liver dysfunction (n = 23). After leaving the hospital all the patients received regular follow- up examination and extended nursing care. Conclusion Usually, the incidence of adverse reactions is higher in patients with advanced HCC after TACE combined with sorafenib treatment. Careful in-hospital observation and extended out-hospital nursing can reduce the incidence of adverse reactions and thus improve the patient’s quality of life.

Objective To investigate feasibility and clinical application value of improved percutaneous transhepatic biliary internal-external drainage (PTBIED).Methods Consecutive patients from April 2007 to April 2010 with malignant obstructive jaundice were diagnosed by medical imaging or pathological confirmation whenever possible.The patients with proximal malignant biliary obstruction and intact inferior common bile ducts ＞ 3 cm in length,and a bilirubin of 70 μmol/L or higher,were included in the experimental group.The control group included patients with low malignant biliary obstruction,and those who met the criteria for the experimental group but refused to receive the altered method of PTBIED.The patients underwent traditional PTBIED in control group.The patients in the experimental group received the procedure as following:according to percutaneous transhepatic cholangiography,a biliary external drainage catheter was modified by adding side-holes.Then under fluoroscopic guidance,the loop tip of the modified biliary drainage catheter was positioned in the inferior common hepatic duct/common bile duct,while the additional side-holes were located in the expanded hepatic duct.Technical success rate,complications,hepatic function and white cell count (WBC) were recorded pre- and post-procedure.All patients were followed-up until death.A t-test was used to compare continuous variable data changes,the Chi-square test was used to compare categorical variable data in two groups,and survival time was assessed using the Kaplan-Meier method.Results Forty-six patients were included in the study,with 21 in the experimental group and 25 in the control group.The procedures were successfully performed in all patients in the two groups.There was no procedure-related death in the two groups.Symptoms were improved similarly after procedures in the two groups.The mean quantity of drained bile per day [experimental group (521 +136) ml/d,control group (606 + 159 ) ml/d,t =1.930,P ＞ 0.05],decrease of the serum total bilirubin after the procedures [ experimental group (87 ± 51 ) μmol/L,control group( 105 ± 66 ) μmol/L ( t =1.061,P ＞ 0.05 ) ] and the median survival time ( experimental group 7.7 months,control group 6.9 months,x2 =0.610,P ＞0.05 ) of the patients showed no statistically significant difference between two groups.The mean WBC amount of patients was higher after the traditional procedure [ ( 10.9 ±5.2) × 109/L] than before the procedure [ (7.8 ±2.9) × 109/L] in the control group ( t =3.606,P ＜ 0.05 ),but the converse change occurred in the experimental group [ pre-procedure (8.2 ± 3.4) × 109/L ],post-procedure [ (7.4 ± 2.6) × 109/L] ( t =2.649,P ＜ 0.05 ).No reflux of duodenal juice was observed in all patients of the experimental group,and 1 patient had infection of biliary tract.The reflux was observed in 11 patients of the control group after conventional PTBIED.Of them,8 patients had infection of biliary tract.Incidence rate of infection of biliary tract in the control group was higher than that in the experimental group( x2 =5.381,P ＜ 0.05 ).Conclusions Improved PTBIED is convenient and feasible,and compared with traditional PTBIED,it can reduce the complications of infection of biliary tract.

Objective To investigate the methods for reconstructing portal vein in liver transplantation patients with grade Ⅳ portal vein thrombosis.Methods Clinical data of 6 patients with grade Ⅳ portal vein thrombosis who underwent liver transplantations were analyzed retrospectively.Different portal vein reconstructing approaches were applied: 4 patients underwent portal vein anastomosis with internal organ varicosis vein (group A),and 2 patients underwent portal vein arterialization (group B). Portal venous flow was monitored by intraoperative ultrasound and postoperative liver function was tested periodically during follow-up.Results In group A,one patient died of celiac infection 2 months post-transplantation.The remaining three patients were followed up for 14-17 months,and their portal veins remained smooth without thrombosis and with mitigated esophageal varicosity.In group B,one patient,with recurrent upper gastrointestinal bleeding,died of celiac infection 47 days after liver transplantation.The patient was followed up for 33 months with satisfactory liver and kidney functions although stomach esophagus varicosity was aggravated.Portal vein blood flow in groups A and B was 1258 ± 345 and 2275 ± 247 ml/min respectively after anastomosis by intraoperative color Dopplar ultrasound monitoring. Aspertate aminotransferase (AST) in group B was significantly lower on the fourth day after liver transplantation,and alanine aminotransferase (ALT) in group B was significantly lower on the 3rd,4th,5th and 6th day after liver transplantation than in group A (all P＜0.05).Serum total bilirubin (TBIL) had no statistically significant difference during the 10 days post-operation (P＞0.05).Conclusion Patients with grade Ⅳ portal vein thrombosis may achieve a satisfactory clinical effect by reconstructing portal vein through anastomosis of donor portal vein with internal organ? varicosis vein.PVA may be associated with early recovery of graft function and may be an effective remedial measure for patients with grade Ⅳ portal vein thrombosis who undergo liver transplantation.

Objective To study the syndrome-drug association rule and the medication law of cough medical cases treated by Xin'an doctors using the data mining method;To provide reference for the treatment of cough.Methods The medical records of Xin'an doctors were collected and summarized.The frequency analysis,topology analysis and Louvain clustering analysis were used to conduct data mining research on syndrome types and prescriptions,and the relationship between syndrome types and prescriptions in cough treatment medical records and the law of drug composition in prescriptions were discussed.Results A total of 525 medical cases were included,and 26 kinds of syndromes were obtained,such as wind-heat invading lung,phlegm-dampness holding lung,lung qi deficiency.There were 243 kinds of Chinese materia medica involved,and 18 kinds of Chinese materia medica were with more than 70 times in frequency.The main properties were cold,warm and mild,and the main tastes were sweet,bitter and pungent,mainly in lung meridian,or the main properties were mild and warm and the main taste was sweet,mainly in spleen and stomach meridian.There were 25 categories in efficacy,which were mainly tonics and heat-clearing medicine.30 core medicines and core medicinal networks were obtained,such as Glycyrrhizae Radix et Rhizoma,Poria and Armeniacae Semen Amarum.The core drug combination of main syndromes of exogenous cough and excess syndrome and deficient syndrome of endogenous cough were obtained by syndrome-drug clustering analysis.Conclusion Xin'an doctors have distinctive characteristics in the diagnosis and treatment of cough diseases,and pay attention to the use of the methods of"strengthening the basic and promoting original qi","nourishing yin and protecting yin","cultivating the soil and promoting gold"and the inheritance of the prescriptions on the basis of cough relieving,phlegm resolving,and syndrome-based treatment.

Objective To study the clinical value of the conventional liver function tests in liver reserve function assessment for large hepatocellular carcinoma.Methods The clinicopathological data of 113 patients with ChildPugh A hepatocellular carcinoma who underwent radical resection with large hepatocellular carcinoma in the Department of Hepatobiliary Surgery of Fuzhou General Hospital of People's Liberation Army from January 2014 to December 2016 were retrospectively analyzed.The patients were divided into two groups according to the recovery of postoperative liver function,which 105 patients recovered well and 8 patients had hepatic decompensation among them.The liver function index of two groups were analyzed.Measurement data with approximately normal distribution were represented by and groups were compared using t test;measurement data with skewed or uneven disstribution were represented by M (range)and group werecompared using Man-Whitney U test;count data were compared using Fisher exact test;risk factors for postoperative liver dysfunction were analyzed using Logistic single factor and multivariate and ROC curve were drawn.Results Preoperative prothrombin time,international standardization ratio,platelet,prealbumin,total bilirubin,alkaline phosphatase,γ-glutamyl transpeptidase comparison between the two groups were statistically significant (Z =-1.983,-2.180,-2.608,-2.007,-3.577,-2.228,-2.575,P ＜ 0.05).Logistic univariate analysis showed that platelet,total bilirubin and prealbumin were the risk factors for the recovery of liver function of radical resection hepatic decompensation with large hepatocellular carcinoma.Logistic multivariate regression analysis showed that preoperative high total bilirubin and low preabumin were independent risk factors of radical resection hepatic decompensation with large hepatocellular carcinoma.Logistic univariate analysis showed that preoperative high total bilirubin and low prealbumin were not risk factors of radical resection liver failure with large hepatocellular carcinoma.The area under the curve of total bilirubin was 0.880,P =0.000,95％ CI:0.808-0.953,the sensitivity was 87.5％,specificity was 84.8％ and the area under prealbumin curve was 0.769,P =0.011,95％ CI:0.648-0.891,sensitivity was 75.2％,specificity was 77.5％ by the ROC curve.The best threshold of total bilirubin and prealbumin after radical resection with large hepatocellular carcinoma were 17.55 μmol/L and 0.18 g/L respectively by the ROC curve.Conclusion The Child-Pugh A patients in radical resection hepatic decompensation with large hepatocellular carcinoma recover well when the preoperative liver function is as follows:total bilirubin ＜ 17.55 μmol/L and prealbumin ≥0.18 g/L.

Objective:To compare the preliminary clinical efficacy of iodized oil embolization and improved microspheres injection embolization in patients with hepatic metastasis from gastrointestinal tumors.Methods:In total 100 patients with hepatic metastasis from gastrointestinal tumors were perspective enrolled from July 2015 to July 2017 in the First Affiliated Hospital of Henan University of Science and Technology. The patients were randomly divided into iodized oil group (45 cases) and microspheres group (55 cases) using random number table. The patients in iodized oil group were firstly received regular perfusion chemotherapy, then underwent the embolization with Iodized oil and epirubicin emulsifier. As for the microspheres group, the patients were received epirubicin and microspheres perfusion and embolization alternately after the same regular perfusion chemotherapy. The post-operation complications were observed. The effective rate of tumor controling at each time point was compared between two groups using χ 2 test. Kaplan-Meier method was used to perform the survival analysis. Log-rank test was used to compare the survival rate between two groups. Results:The operation was successfully performed in all the patients, without severe complications found. The follow up time was 18.7±3.4 months. The tumor control rates of 1, 3, 6, 12 and 24 months after operation in the iodine oil group were 71.1% (32/45), 68.9% (31/45), 51.1% (23/45), 28.9% (13/45), 6.7% (3/45) respectively; while in microspheres group were 90.9% (50/55), 89.1% (49/55), 72.7% (40/55), 49.1% (27/55), 23.6% (13/55), respectively. The tumor control rate in microspheres group was superior to that in iodine oil group ( P<0.05). The median lifetime was 12 months and 17 months for iodine oil group and microspheres group respectively, with significant difference (χ2=8.238, P=0.004) found between two groups. As for the microspheres group, the liver abscess was found in one week after operation in 4 patients, who were cured after drainage and anti-inflammatory treatment. Conclusion:The improved microspheres injection embolization is an effective method for the hepatic metastasis from gastrointestinal tumors and superior to iodized oil embolization in tumor controlling, which may prolong the median survival time.

Objective To investigate the feasibility of liver transplantation in the treatment of inoperable hilar cholangiocarcinoma. Methods The clinical data for 3 patients with unresectable hilar cholangiocarcinoma who underwent liver transplantation in the Department of Hepatobiliary Surgery of Fuzhou General Hospital of People's Liberation Army from January 2006 to December 2012 were retrospectively analyzed. The patients were followed up by phone, outpatient service, and hospitalization. The starting point of the follow-up was the operation date. The patients death was the end point. The clinical and pathological features, postoperative survival, tumor recurrence, and prognostic factors were observed. The follow-up deadline was December 2017. Results All 3 patients underwent classical orthotopic liver transplantation using retrograde perfusion through inferior vena cava and no perioperative deaths occurred. All 3 patients were followed up for 10 to 132 months. During the follow-up period, of 1 patient who died of tumor recurrence, the pathological TNM stage was T4a N1 M0, and both had Union for International Cancer Control stage Ⅳa, and the tumor-free survival time was 3 months, and the survival time was12 months. Of 1 patient who died of other causes, the pathological TNM stage was T3N1 M0, and both had Union for International Cancer Control stage Ⅲ, and the tumor-free survival time was 12 months, and the survival time was12 months. One case as of the end of follow-up, the patient has survived for 132 months, the pathological TNM staging was T2a NOM0, and both had Union for International Cancer Control stage Ⅱ. Conclusions Lymph node positive and high pathological TNM stage were poor prognosis factor for hilar cholangiocarcinoma who underwent liver transplantation. Patients with early hilar cholangiocarcinoma who don't have lymph node metastasis are expected to benefit from liver transplantation.

Objective To evaluate the efficacy and safety of 125I seed implantation combined with transcatheter arterial chemoembolization (TACE) in the treatment of primary hepatocellular carcinoma (PHC).Methods 156 patients who had unresectable PHC seen from January 2012 to December 2015 in our unit were randomly assigned into the study group (76 patients) and the control group (80 patients).The study group of patients were treated with intra-hepatic implantation of 125I seed + TACE,and the control group of patients were treated with three dimensional conformal radiotherapy (3DCRT) + TACE.After ra diotherapy the two groups of patients underwent 2 to 3 times TACE treatment.The biomarker levels were compared between the two groups before and after treatment,and the short-term efficacy,clinical safety and 1 year survival rates were assessed.Results The success rates of the treatment for the two groups were both 100％.Patients in the study group were implanted with 1 016 125I seeds,with an average of (13.7 ± 2.5) seeds per patient.The two groups had no significant difference on short-term efficacy (P ＞ 0.05).The DCR,ORR and 1 year survival rates in the study group were 63.2％,92.1％,55.5％,and they were 61.3％,90.0％,58.1％ in the control group,respectively.There were no significant differences between these 2 groups (P ＞ 0.05).At 1 month after treatment,the two groups had no significant differences onAFP,IGF-Ⅱ and IGFBP-2 [(5.08±0.85) μg/L vs.(5.12 ±0.79) μg/L,(4.77 ±0.58) μg/L vs.(4.86 ±0.53) μg/L,(4.98 ±0.67) μg/L vs.(5.04 ±0.71) μg/L] (P＞0.05).There were no signif-icant differences on chemotherapy drug toxicities between the two groups (P ＞ 0.05).The incidence of radi-ation hepatitis in the study group was 1.3％,while the incidences of radiation dermatitis,hepatitis,gastriculcer in the control group were 5.0％,8.8％,2.5％,respectively.The radiation dermatitis incidence inthe study group was significantly lower than that of the control group (P ＜ 0.05).In the study group,2 pa-tients (2.6％) had their 125I seed slightly moved.Conclusion The short-term efficacy of 125I seed implan-tation plus TACE in the treatment of PHC is no less than the 3DCRT + TACE regimen,but with less radia-tion side effects.