2.Purification and Properties of Neutral Protease from Bacillus Subtilis ZC-7
Cong ZHAO ; Min ZHANG ; Jian-Ling WANG ; Lian-Xiang DU ; Xiang-Bin YIN ;
China Biotechnology 2006;0(10):-
Bacillus subtilis ZC-7 was obtained by implantation with N+ ions beam to B.subtilis AS1.398,and compared with the AS1.398 neutral protease,the enzyme activity of ZC-7 neutral protease was about 1 timeshigher in previous research.A neutral protease was purified from the culture of B.Subtilis ZC-7 by the procedures including amoninium sulfate precipitation,ultrafiltration,DEAE-Sepharose Fast Flow chromatography and Sephadex G-75 chromatography.By multi-step purification,the ZC-7 neutral protease was purified to 78.5 folds and its yield was 27.7%,at last,the specific activity of ZC-7 neutral protease was up to 4.1?105U/mg.Analysed by SDS-PAGE,the purified protease has shown a molecular mass of about 42kDa.The Km for casein hydrolysis was 3.67?10-3?g/ml and the Vmax was 12.21?g/min.The optimum pH and temperature forhydrolysis of casein were 7.0 and 55℃,respectively.This protease was stable up to 40℃ within the pH range of 6.5 and 8.0.EDTA,isopropanol and alcohol nearly inhibited its activity while some ions such as Ca2+,Mg2+,Fe3+ can improve its activity.In addition,it could resist 1 mol/L H2O2.
3.Determination of paclitaxel prodrug in SD rat plasma by LC-MS/MS and its application in preclinical pharmacokinetic studies
Guo-cai WANG ; Xiang-yi WANG ; Cong-cong XIAO ; Jian-peng HUANG ; Meng YU ; Jiu-ming HE
Acta Pharmaceutica Sinica 2022;57(9):2798-2804
A fast and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of prodrug of paclitaxel (Pro-PTX) and paclitaxel (PTX) in rat plasma was developed. The plasma samples were subjected to protein precipitation with acetonitrile (0.1% formic acid), and then separated by LC with an Ultimate AQ-C18 column (50 mm × 3.0 mm, 3 μm) and acetonitrile-1 mmol·L-1 ammonium formate (containing 0.1% formic acid) as the mobile phase. Multiple reaction monitoring (MRM) scanning mode was used to detect the ion responses
4.Isolation and identification of human periodontal ligament stem cells in vitro.
Tao SHEN ; Hui-jun CHANG ; Cong-xiang JIAN ; Yan-chun YANG ; Ji-xiang ZHOU
West China Journal of Stomatology 2011;29(1):71-78
OBJECTIVETo isolate and identify human periodontal ligament stem cells (PDLSC) by improved methods and assess the characteristics of PDLSC ex vivo.
METHODSThe periodontal ligament cells were obtained from the healthy impacted third molars and teeth extracted for orthodontic purposes and used to isolate PDLSC by limiting dilution assay. PDLSC were cultured and expanded in alpha-MEM supplemented with 10% FBS. Colony-forming assay, immunohistochemistry, flow cytometry, osteogenic and adipogenic induction were used to identify PDLSC.
RESULTSThe obtained cells had high colony-forming efficiency and were positive staining for vimentin and negative for pancytokeratin. Flow cytometry revealed that the isolated cells were positive for STRO-1 and CD146 antibodies and most were in the G0/G1 phase of cell cycle. Under specific conditions, they could differentiate to the osteoblast and adipocyte lineages in vitro.
CONCLUSIONLimiting dilution assay is an effective method to isolate PDLSC and the single-cell-derived colonies demonstrate the properties of stem cells in vitro.
Cell Differentiation ; Cell Separation ; Humans ; In Vitro Techniques ; Osteoblasts ; Periodontal Ligament ; Stem Cells
5.Dynamic image-pathological investigation on MR diffusion weighted imaging in rabbit liver VX_2 tumor model
You-Hong YUAN ; En-Hua XIAO ; Jian-Bin LIU ; Zhong HE ; Ke JIN ; Cong MA ; Jun XIANG ; Jie-Hua XIAO ; Wei-Jian CHEN ; Zi-Wen PENG
Chinese Journal of Radiology 2001;0(01):-
Objective To investigate the value and dynamic characteristics of diffusion-weighted imaging(DWI)in rabbit liver VX_2 tumor model,with correlation of pathology.Methods Forty New Zealand rabbits were included in the study and VX_2 tumor piece was implanted directly into the liver after laporotomy.Fiffteen had two intrahepatic implants while twenty-five had one implant.DWI was performed on the seventh,fourteenth and twenty-first day after implantation,while routine T_1WI and T_2WI sequences were done on the seventh and fourteenth day.Ten VX_2 tumor samples were studied by pathology.Results The lump detection rates on the seventh day after implantation of DWI,T_1WI and T_2WI were 78.7%(37/47), 10.7%(3/28)and 53.6%(15/28)respectively with statistical significance(x~2=32.61,P
6.A dynamic study of correlation between the MR diffusion weighted imaging findings and the expression of proliferation-related and metastasis-related genes in rabbit models of liver VX,tumor before and after chemoembolization
You-Hong YUAN ; En-Hua XIAO ; Jian-Bin LIU ; Zhong HE ; Ke JIN ; Cong MA ; Jun XIANG ; Jie-Hua XIAO ; Wei-Jian CHEN ;
Chinese Journal of Radiology 2001;0(07):-
0.05).Conclusions The potential of VX_2 tumor infiltration and metastasis will decrease after chemoembolization in the long-run,while the proliferation ability will increase within a short period of time. The ADC may reflect the proliferation of VX_2 tumor cells.
7.The relationship between hemostatic changes in liver cirrhosis patients with different degrees of liver lesions in reference to Child-Pugh scores.
Yu-Long CONG ; Yu-Xiang WEI ; Li-Wen ZHANG ; Zong-Jian YIN ; Jie BAI
Chinese Journal of Hepatology 2005;13(1):31-34
OBJECTIVETo investigate the relationship between hemostatic changes in liver cirrhosis patients with different degrees of their liver lesions.
METHODSForty-three patients (35 men, 8 women; age: 25 to 71 yr) with liver cirrhosis were divided into three subgroups (A, B, and C) on the basis of Child-Pugh classification. Among the patients, 13 were classified as Child-Pugh class A, 15 were class B, 15 were class C. 16 healthy individuals served as controls. A series of hemostatic tests and parameters including prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fib), factors II, V, VII, VIII, IX, X, vWF assay, antithrombin-III (AT-III), protein C (PC), D-dimer, tissue plasminogen activator antigen (t-PA), plasminogen activator inhibitor activity (PAI) were performed on 43 patients and the 16 healthy controls.
RESULTSPT and APTT were progressively prolonged from A to B and then to C. In comparison to the controls there was a significant difference. Fibrinolytic activity and the activities of factors II, V, VII, IX, X were progressively decreased from A to B and then to C. In comparison to the controls there was a significant difference . AT-III and PC activity were progressively decreased from A to B and then to C. In comparison to the controls there was a significant difference. D-dimer and t-PA-antigen were progressively increased from A to B and then to C. In comparison to the controls there was significant difference. PAI activity did not display significant changes in the four groups.
CONCLUSIONWe found that there is a close relationship between the severity of cirrhosis and the hemostatic changes. Because the deterioration of the coagulation function and increasing fibrinolytic activity parallel the severity of liver cirrhosis, adequate treatment for cirrhotic bleeding should not only correct the coagulation defects, but also lower the increased fibrinolytic activity.
Adult ; Aged ; Antithrombins ; metabolism ; Blood Coagulation Factors ; metabolism ; Female ; Fibrinogen ; metabolism ; Hemostasis ; Hepatitis B, Chronic ; blood ; complications ; Humans ; Liver Cirrhosis ; blood ; diagnosis ; etiology ; Male ; Middle Aged ; Prothrombin Time ; Severity of Illness Index
8.Treatment of calcaneal fractures by fixation of Kirschner needle and thread cancellous bone screw through sinus tarsi interstice.
Cong-Xiang LIN ; Zheng-Yang SHI ; Yi-Min XU ; Meng YANG ; Jia-Lin WANG ; Xiao-Jian CHEN ; Wei ZHANG ; Shao-Xin ZHI
China Journal of Orthopaedics and Traumatology 2014;27(7):551-554
OBJECTIVETo explore the effect of fixation of Kirschner needle and thread cancellous bone screw through the sinus tarsi interstice for the treatment of calcaneal fractures.
METHODSFrom January 2009 to December 2012,20 patients with calcaneal fracture were treated by minimally invasive Kirschner wire and threaded cancellous bone screw fixation and bone graft,including 12 males and 8 females with an average age of 39 years old ranging from 21 to 65. Among them, 8 cases were left foot, 12 were right foot. According to Sanders's classification, 8 cases were type II, 10 cases were type III, 2 cases were type IV.
RESULTSAll patients were followed up from 6 to 16 months with an average of 12 months. The incision were healed. Böhler angle were increased from preoperative (17.75 +/- 4.22) degrees to postoperative (26.85 +/- 7.37) degrees (t = 4.308, P = 0.000). Gissane angle were reduced from preoperative (137.05 +/- 24.91) degrees to postoperative (113.75 +/- 13.17) degrees (t = 7.083, P = 0.000). At 3 months after operation, the scores of AOFAS were 85.50 +/- 7.99; the results were excellent in 5 feet and good in 11 feet, fair in 3 feet, and poor in 1 foot.
CONCLUSIONMinimally invasive fixation of Kirschner needle and thread cancellous bone screw fixation is a simple operation, it can get reliable fixation, easy to remove, low cost, less postoperative complications, and it is a good treatment of calcaneal fracture.
Adult ; Aged ; Bone Screws ; Bone Wires ; Calcaneus ; injuries ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged
9.Error analysis and experience summary in setting up the retroperitoneal cavity for peritoneoscopy
jian Xiao CUI ; cheng Gui CONG ; xiang Qing YAO ; quan Bo LI
Tianjin Medical Journal 2017;45(12):1300-1303
Objective To explore errors and their causes in setting up the retroperitoneal cavity for peritoneoscopy. Methods The clinical data of 450 patients who were performed the laparoscopic surgery in our hospital from May 2009 to December 2016 were collected. According to the trocar puncture points, patients were divided into lumbar group (n=193) and iliac flap group (n=276). The problems were summarized and analyzed in the process of setting up the retroperitoneal cavity. Results The mistakes existed in setting up the retroperitoneal cavity including peritoneum rupture (10 cases), error in balloon expansion clearance (5 cases), homemade balloon rupture and fall off (7 cases), poor position of puncture port (34 cases), bleeding of puncture channel (6 cases), leaking around the trocar and subcutaneous emphysema. After peritoneal patching, re-establishment of the expansion of the gap, adjusting the trocar position and other appropriate measures for treatment, the operations were successfully in 450 patients. Conclusion We should choose the appropriate method for building cavity according to different conditions of patients, and know well the anatomy of the peritoneal cavity. All details should be emphasized in the process of building cavity to reduce the occurrence of errors.
10.Impact of uterine fibroid embolization with danazol alginate microsphere on ovarian function and subsequent pregnancy
Cheng-Zhi LEI ; Yang XIANG ; Guo-Kun AO ; Li LI ; Ying-Chang SHI ; Yi-Rong BAO ; Cong-Jian XU ; Hong HONG ; Jing-He LANG
Chinese Journal of Obstetrics and Gynecology 2000;0(10):-
0.05).Conclusions There is no obvious effect of danazol alginate microspheres used for uterine arterial embolization on ovarian function in rabblits.After UAE some animals are able to achieve pregnancies,while harmful effects are observed on short term pregnant rate.