Bladder tissue engineering includes three components, namely cell scaffold, seeding cells and message factor. Seeding cells is essential to construct tissue engineering organs and is the native material of organ reconstruction. It is important to find a seeding cell which can not only provide excellent biocompatibility, but also can generate enough cells in work of organ reconstruction. At present, there are some commonly used seeding cells, for example, urinary bladder transitional epithelial cells,smooth muscle cells, coculture of smooth muscle cells and transitional epithelial cells, mesenchymal stem cells. Among the total,coculture of smooth muscle cells and transitional epithelial cells and mesenchymal stern cells are better than the others. The current problems of seeding cells are as follows: ①The mature cells that have been completely differentiated are limited in source and immunological rejection. ② The mature cells that have been completely differentiated during the differentiated only can generate in the limited passage culture procedure and they will age and die out of the passage maximum; in addition, the dedifferentiation also exists; ③further analysis should focus on the coculture of smooth muscle cells and transitional epithelial cells and mesenchymal stem cells into normal bladder.

Efforts on directed evolution of sortase A to optimize its catalytic properties have been undertaken and shown the promise. To facilitate screening of sortase A mutants with expected catalytic properties, a novel ligation efficiency monitoring system, including reporter substrates GFP-LPETG and GGGYK-Biotin, was developed. GFP-LPETG, wild type sortase A, and a recently reported high activity sortase A mutant were prepared recombinantly from Escherichia coli BL21 (DE3). Taking advantage of the newly designed reporter system, the ligation efficiency catalyzed by wild type and mutant form of sortase A could be sensitively monitored via SDS-PAGE directly. Consistent with previous report, the mutant sortase A displayed much higher catalytic activity compared to wild type enzyme, indicating the new reporter system is easily and fast handled and sensitive. The application of this reporter system into systemic screening will facilitate future directed optimization of sortase A.
Aminoacyltransferases ; genetics ; metabolism ; Bacterial Proteins ; genetics ; metabolism ; Biocatalysis ; Biotin ; Cysteine Endopeptidases ; genetics ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Genes, Reporter ; Ligation ; Mutant Proteins ; genetics ; metabolism

Background and purpose:The clinical relevance of HBV infection with respect to diffuse large B cell lymphoma(DLBCL) patients and immune patterns of T lymphocyte subsets during chemotherapy remains unclear. This study aimed to identify the characteristics of T-cell mediated immunity in DLBCL patients with HBV infection, thereafter, to explore the possible cell-mediated immune mechanisms of HBsAg positive HBV infection on the survival of DLBCL. Methods:A total of 294 newly diagnosed DLBCL patients were enrolled in this cohort study. Four-color flow cytometric method was used to enumerate the absolute number of CD3+, CD4+, CD8+T lymphocytes and the CD4+/CD8+ratio in peripheral blood samples, at the onset of disease, 2-4, 4-6 and 6-12 months after the initiation of chemotherapy, individually. Results:The absolute number of CD3+, CD4+, CD8+T lymphocytes in both groups were similar at the onset of disease;the count of CD4+lymphocytes was lower in HBsAg positive group during 2 to 4 months after the initiation of chemotherapy, compared with that in the HBsAg negative group. During 4 to 12 months after chemotherapy, the CD4+/CD8+ratio in peripheral blood samples was significantly lower in HBsAg positive group. Conclusion:For newly diagnosed DLBCL patients who received chemotherapy, the dynamic nature of cell mediated immune response was characterized as a low counts of CD4+T lymphocyte during the ifrst several cycles of chemother-apy followed by a decreased circulating CD4+/CD8+ratio. Depressions of cell immunity after chemotherapy in HBsAg positive DLBCL patients were greater and prolonged.

Objective To evaluate the clinical value of MSCT in the diagnosis of unexplained aseites.Methods 113 patients with unexplained ascites were retrospectively reviewed by CT、clinical data and continous observation.Results Large ascites in 50 cases(44.3%),moderate amount aseites in 10 cases(8.8%),small ascites in 53 cases(46.9%);Parietal peritoneum changed in 51 cases(45.1%);Mesentery changed in 44 cases(38.9%);Grerter omentum changed in 20 eases(17.7%);Enlargement of lymph nodes in 67 cases(59.3%).Conclusion Malignant tumor was the most common cause of unexplained ascites.MSCT could help in identifying tumors and the tumor lesion,forecast malignant ascites,and had great value in etiology and diagnosis of ascites.

Objective To study the factors affecting the activities of daily life (AOL) of patients after the first stroke of cerebral infarction in order to formulate the intervention strategy for improving the capability of ADL of patients. Method A total of 149 patients with the first stroke of cerebral infarction admitted from Oct. 2008 to Dec.2008 were enrolled in this study. The demographics of patients, cerebral infarction risk factors, apopletic score as per National Institute of Health stroke scale (NIHSS), white blood cell count (WBC) and plasma glucose (PG) were recorded on the first day of admission, and many other laboratory examinations were done on the next morning. The occurrences of infection in lung and urinary tract, and atrial fibrillation were recorded during hospi-talization. The NIHSS score and score of Glasgow Coma Scale (GCS) were taken within 24 hours after admission, on the 14th day and at the end of the third month after cerebral infarction respectively. Barthel index (BI) was taken in the second week as well as at the end of third month by follow-up in the OPD or by telephone. The correlation analysis and multiple linear stepwise regression analysis were used to find the risk factors. Results The PG level, WBC count and NIHSS score were independently associated with ADL in the second week as well as at the end of the third month after cerebral infarction. Besides, the urinary tract infection during hospital stay was also independently associated with ADL at the end of the third month after cerebral infarction. Conclusions The plasma glucose level, WBC count, NIHSS score and urinary tract infection are the risk factors. Positive measures should be taken to control these risk factors so as to improve the capability of ADL of the patients after cerebral infarction.

Metabolic syndrome is a clinical syndrome characterized by the co-existence of obesity, hyperglycemia, hypertension, dyslipidemia and other metabolic disorders.At present, the incidence of metabolic syndrome is increasing year by year.Metabolic syndrome is more common among the elderly, seriously threatening the quality of life in the elderly.Tea, as one of the most popular beverages in the world, has been found to have effects on lowering the levels of blood pressure, blood glucose and lipid and so on.This review summarizes the effects and possible mechanisms of tea drinking on metabolic syndrome and its components in the elderly.

Objective To investigate the efficacy of double-“ U” embedding and pursestring suture and binding pancreaticojejunostomy for the prevention of pancreatic fistula.Methods The retrospective cohort study was adopted.The clinical data of 208 patients who underwent pancreaticojejunostomy at the Hunan Provincial People's Hospital from March 2011 to March 2015 were collected.Of 208 patients,106 patients undergoing double-“ U” embedding and pursestring suture and binding pancreaticojejunostomy were allocated into the double-“ U” group and 102 patients undergoing Child pancreaticojejunostomy were allocated into the Child group.Observation indicators included (1) surgical effects:anastomosis time,postoperative pancreatic leakage,duration of hospital stay,(2) follow-up situations.The follow-up using telephone interview and outpatient examination was performed to detect postoperative long-term complications and recovery of patients by abdominal ultrasound or computed tomography (CT) at every 6 months postoperatively up to September 2015.Measurement data with normal distribution were represented as x ± s and comparison between groups was analyzed by t test.Count data were analyzed using the chi-square test.Results (1) Surgical effects:208 patients underwent successful surgery without occurrence of death.The anastomosis time was (13.0 ± 1.5) minutes in the double-“ U” group and (20.0 ± 1.6) minutes in the Child group,with a statistically significant difference between the 2 groups (t =4.713,P ＜ 0.05).Two patients in the double-“ U” group were complicated with grade A of pancreatic leakage,including 1 of 36 patients with normal pancreatic remnant and 1 of 70 patients with fibrotic pancreatic remnant.Nine patients in the Child group were complicated with pancreatic leakage,including 6 in grade A,1 in grade B and 2 in grade C,and there were 6 of 33 patients (4 in grade A,1 in grade B,1 in grade C) with normal pancreatic remnant and 3 of 69 patients (2 in grade A,1 in grade C) with fibrotic pancreatic remnant.There were statistically significant differences in the pancreatic leakage between the 2 groups and among the patients with normal pancreatic remnant in the 2 groups (x2 =2.951,4.994,P ＜ 0.05).The duration of postoperative hospital stay was (13.5 ± 1.2)days in the double-“U” group and (15.7 ± 2.6)days in the Child group,with a statistically significant difference (t =1.011,P ＜ 0.05).No readmission in the 2 groups occurred.(2) Followup situations:91 of 106 patients in the double-“U” group were followed up for 6-54 months with a median time of 30 months.During the follow-up,8 patients were dead,12 patients didn't undergo reoperation due to multiple metastases in the liver,lung and greater omentum,4 and 4 patients were respectively complicated with relapsing pancreatitis and refluxing cholangitis,and other patients had good conditions without the occurrence of diabetes,diarrhea,indigestion and hypopancreatism.Eighty-eight of 102 patients in the Child group were followed up for 6-54 months with a median time of 25 months.During the follow-up,10 patients were dead,11 patients didn't undergo reoperation due to multiple metastases in the liver,lung and greater omentum,6 and 6 patients were respectively complicated with relapsing pancreatitis and refluxing cholangitis,and other patients had good conditions without the occurrence of diabetes,diarrhea,indigestion and hypopancreatism.Conclusion Double“U” embedding and pursestring suture and binding pancreaticojejunostomy for the prevention of pancreatic fistula can reduce the suture time,incidence of pancreatic leakage and duration of postoperative hospital stay,and it is especially suitable for the patients with normal pancreatic remnant.

The viewpoint of treating gallbladder and combinedtreatment of liver and gallbladder for liver diseasse isproposed and the therapeutic project of intravenousdrip of Mixture of Radix Bupleuri plus Radix SalviaeMiltiorrhizae is formulated.Sixty eases of chronic hep-atitis of damp—heat of liver—gallbladder with block-age of collaterals by stagnant blood were thus treated.Results revealed the relief of jaundice,decrease of en-zyme and inhibition of viral replication in treatinggroup are all better than the western drug controlgroup.

Objective To evaluate a method of fast detection of the hematopoietic progenitor cell (HPC) in peripheral blood samples and explore for an appropriate cutoff value in prediction of adequate CD34+ cell in apheresis concentrate.Methods Peripheral blood samples and apheresis concentrate samples were collected from 27 auto-PBSCT patients receiving chemotherapy plus G-CSF mobilization (chemo group) and 17 patients receiving G-CSF alone (non-chemo group).CD34+ cell counts were determined by flow cytometry according to ISHAGE guideline and HPC counts were detected using Sysmex XE-2100 automatic hemocyte analyzer.The correlation between HPC and CD34+ cell counts in peripheral blood samples and apheresis concentrates were analyzed.Receiver operating characteristic (ROC) curves was used to determine the cutoff value in prediction of adequate CD34+ cell in apheresis concentrate.Results CD34+ cell counts in peripheral blood samples can be estimated by HPC counts (r =0.711,P =0.000,r =0.656,P =0.004).CD34+ cell counts =-0.829+0.648×HPC counts (in chemo group) or 45.033+0.460×HPC counts (in non-chemo group).HPC counts in the peripheral blood of auto-PBSCT patients were highly correlated with the CD34+ cell yield (r =0.602,P =0.001),CD34+ cell counts =1.106+0.046×HPC counts.When HPC in peripheral blood was ≥85/μl,the prediction of adequate CD34+ cells in the yield of apheresis (≥5×106/kg body weight) would have a sensitivity of 78 ％ and a specifity of 82 ％.Conclusion HPC counts in peripheral blood samples in auto-PBSCT patients can be used to determine the optimal time of apheresis and be used as a good marker to predict the stem cell in the yield.

Background and purpose:Positron emission tomography-computed tomography (PET/CT) is playing an increasingly important role in the diagnosis, therapy and follow-up of lymphoma patients. This study aimed to explore clinical and pathological features and bone marrow infiltration status in lymphoma patients with diffused high bone marrow glucose uptake on18F-FDG PET/CT.Methods:It was a retrospective study. Bone marrow infiltration status, pathological and clinical data from 62 cases of pathologically diagnosed lymphoma and diffused high bone marrow glucose uptake were analyzed.Results:Distribution of histopathological subtype in those cases was in accordance with that in previously reported Chinese lymphoma patients. Significant difference was demonstrated in standard uptake value (SUV) between pa-tients with aggressive and indolent histopathological subtypes (8.43vs 5.38,P=0.048), patients with and without B symp-toms (8.30vs 5.72,P=0.033), and patients with and without bone marrow infiltration (8.78vs 6.96,P=0.020). 32 patients were diagnosed as “bone marrow infiltration” by bone marrow biopsy. There was significant difference in histopathologi-cal subtype distribution between patients with and without bone marrow infiltration (P=0.001). In patients with bone marrow infiltration, there were higher proportions of mantle cell lymphoma, nodal marginal zone B cell lymphoma, Burkitt’s lym-phoma and anaplastic large cell lymphoma. In contrast, patients without bone marrow infiltration suffered more from diffuse large B-cell lymphoma, peripheral T cell lymphoma, enteropathic T cell lymphoma and extranodal NK/T-cell lymphoma (nasal type). False positive results in bone marrow glucose uptake may be caused by fever or anemia.Conclusion:Diffused high bone marrow glucose uptake on18F-FDG PET/CT should be evaluated in combination with the uptake values, clinical features and histological subtypes, to minimize the misdiagnosis and to better guide staging and therapy of lymphoma.