Adolescent ; Adult ; Angiofibroma ; radiotherapy ; Humans ; Male ; Nasopharyngeal Neoplasms ; radiotherapy ; Neoplasm, Residual ; Postoperative Period ; Young Adult

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.

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 m/z 983.4→415.2 (Pro-PTX), m/z 854.4→286.1 (PTX) and m/z 808.3→527.2 (Docetaxel, internal standard) by using a triple quadrupole tandem mass spectrometer with electrospray ionization source and positive ion mode. The method validation results show that the linear ranges of Pro-PTX and PTX in plasma were 2.00-500 ng·mL^-1 (r > 0.99) and 4.00-1 000 ng·mL^-1 (r > 0.99), the lower limit of quantification was 2.00 ng·mL^-1 and 4.00 ng·mL^-1, respectively; the quality control samples with low, medium and high concentrations of Pro-PTX and PTX were within the batch, the relative standard deviation (RSD) between batches were all less than 9%; the relative deviation (RE) was within the range of ± 9%; In the stability test, both Pro-PTX and PTX in plasma were stable under various storage conditions. The method was sensitive, specific, and reproducible, and was suitable for the pharmacokinetic study of Pro-PTX in rats. Animal experiments were approved by the Ethics Committee of Laboratory Animal Management and Animal Welfare, Institute of Materia Medica, Chinese Academy of Medical Sciences (No.: 00003552).

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

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

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.

OBJECTIVETo investigate the influence of partial deletions in the AZFc region of the Y chromosome on spermatogenesis.

METHODSWe selected 9 sequence tagged sites (sY1258, sY1291, sY254, sY255, sY1201, sY1206, sY1161, sY1197 and sY1191) in the AZFc region of the Y chromosome, with ZFX/ZFY and SRY (sY14) as the interior control. We amplified by multiplex PCR the DNA of 160 patients with azoospermia or severe oligozoospermia that showed no microdeletion of the Y chromosome (the case group) and another 76 males with normal fertility (the control group). For the individuals suspected of DAZ gene deletion, we detected the single nucleotide polymorphism sites (SNPs) in the four copies of the DAZ gene by single nucleotide variation (SNV) analysis to determine the types of DAZ copy deletion.

RESULTSIn the case group, there were 10 cases of gr/gr (sY1291) deletion (6.3%), 14 b2/b3 (sY1191) deletion (8.8%), 1 sY1291,sY1197 deletion (0.6%), 1 b1/b2 deletion (0.6%) and 1 b1/b3 deletion (0.6%), while in the control group, there were 4 cases of gr/gr deletion (5.3%) and 4 b2/b3 deletion (5.3%). SNV analysis showed DAZ1/DAZ2 deletion in all those with gr/gr and those with b1/b3 deletion, DAZ3/DAZ4 deletion in those with b2/b3 deletion, and DAZ-SNV sY587 deletion in 1 case of sY1291, sY1197 deletion, but no DAZ deletion was found in 1 case of b1/b2 deletion.

CONCLUSIONB2/b3 (sY1191) and gr/gr (sY1291) deletions are genomic polymorphisms and quite common in the normal Chinese population; while b1/b2, b1/b3, and sY1291, sY1197 deletions may be high risk factors of dyszoospermia.

Case-Control Studies ; Chromosomes, Human, Y ; Humans ; Male ; Oligospermia ; genetics ; Sequence Deletion ; Spermatogenesis

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

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

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.