In this paper,300 bacteria strains and 31fungi strain were isolated from Qinghai plateau.The numbers of cellulose degradation organisms in soil is 2.6?10 5/g. A strain Trichoderma koningii No.0143 which produces cellulase was isolated from 11 fungi in the east area in QingHai,its FPA activity was 15u/g. It can be used in enzymatic feed.

OBJECTIVETo compare the changes of nucleus pulposus after in vitro culture of rabbit whole intervertebral disc and spinal motion segment.

METHODSTwenty-one New Zealand white rabbits which were randomly divided into organ group with 8 rabbits and segment group with 13 rabbits. Fifty intervertebral discs and 50 spinal motion segments were harvested respectively under aseptic conditions from two groups. These specimens were maintained in organ culture with hyperosmotic media (410 mOsm/kg), then 10 discs of the two groups were observed respectively by HE staining, immunohistochemistry of collagen type III, proteoglycan content and cells viability of nucleus pulposus before culture and at 3, 7, 14, 21 days after culture.

RESULTSHE staining showed the intervertebral disc tissue structure remained intact after culture of 21 days organ group and 14 days segment group,but there was severely degenerated of 21 days segment group. The intensity value of type II collagen immunohistochemical staining in the nucleus pulposus were not changed significantly between 21 days organ group and 14 days segment group (P > 0.05), but the staining of segment group at 21 days became shallower, there was significant difference compared with before each time points and organ group at 21 days (P < 0.05). PAS/AB staining of proteoglycan of nucleus pulposus showed that there were not decrease of tinting strength of two groups within 7 days, but the strength weakened slightly of two groups at 14 days, and the tinting strength became weaker at 21 days segment group, the change is more obvious than the organ group. The intensity value of fluorescence staining of nucleus pulposus cells was not changed significantly within 7 days of two groups (P > 0.05), the intensity value decreased slightly at 21 days organ group and 14 days segment group, but there were no significant difference compared with before time points (P > 0.05) however at 21 days segment group the intensity decreased as cells viability of nucleus pulposus decreased,and there was a significant difference compared with before each time points and organ group at 21 days (P < 0.05).

CONCLUSIONIt is not obviously degenerated of the discs of organ group cultured within 21 days and segment group cultured within 14 days, but there was significant degeneration of the intervertebral disc of segment group after cultured 21 days, so the rabbit spinal motion segment can be used on research about the biomechanics of intervertebral disc as a vitro experimental model within 14 days.

Animals ; Collagen Type II ; analysis ; Female ; Immunohistochemistry ; Intervertebral Disc ; chemistry ; pathology ; Male ; Organ Culture Techniques ; Rabbits

Objective To describe the hemodynamic changes during piggyback liver transplantation (PBLT), and to analyze the hemodynamic correlation with various degrees of cirrhosis according to Childpugh classification. Methods Between March 1999 and June 2004, 180 patients underwent PBLT procedure in our institution, and 95 cases were selected and divided according to Child classification. The intraoperative hemodynamics of different time points were retrospectively analyzed, including mean artery pressure (MAP), heart rate (HR), central vein pressure (CVP) and mean pulmonary artery pressure (MPAP). Results Hemodynamic changes were minimal before and during anhepatic phase in all the patients. At reperfusion, a hemodynamic disturbance occurred featured by decrease of MAP and increase of MPAP. Comparison between different Child class showed that in the Child C group, MAP were lower and HR were higher before new liver phase, while CVP and MPAP were higher during new liver phase. Conclusion Hemodynamic changes were minimal before and during anhepatic phase for PBLT, while they were more severe during reperfusion, and they also correlates with the different Child class before transplantation. The more severe of the cirrhosis before transplantation according to Child classification, the greater hemodynamic changes during the operation.

OBJECTIVEThis study was designed to detect the expression of bcl-2 and p53 proteins in colorectal carcinomas and to determine their association with the patient survival and stage of the diseases.

METHODSImmunohistochemistry method was used to detect the expression of bcl-2 and p53 proteins in 93 cases of colorectal carcinoma. The stain results were obtained by analyzing the clinic-pathological characteristics of patients.

RESULTSFifty-seven percent (53/93) of the colorectal carcinomas were bcl-2 protein positive. The positive rate of bcl-2 protein in lymph node involvement cases was lower (15/37) than the cases without node involvement (38/58, P<0.01). The positive rate of p53 protein was 43% (40/93) in colon-rectum carcinomas. No significant correlation was observed between p53 protein expression and clinic-pathological manifestations (P>0.05) but the survival was significantly worse (P=0.0001) in the p53 protein positive cases. Neither bcl-2 nor p53 alone was correlated with stage of the disease. When combined bcl-2/p53 status was analyzed, a group with bcl-2(+) and p53(-) had the best prognosis. This group was significantly associated with earlier Dukes' stages (P=0.1763). In multivariate Cox regression analysis, lymph node involvement and p53 protein expression were two independent factors correlated with survival time.

CONCLUSIONThe expression of bcl-2 and p53 represent biological characteristics of colorectal carcinomas. Assessment of both bcl-2 and p53 status may be valuable in predicting the prognosis of patients.

Biomarkers, Tumor ; metabolism ; China ; epidemiology ; Colorectal Neoplasms ; diagnosis ; metabolism ; mortality ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Risk Assessment ; methods ; Risk Factors ; Survival Analysis ; Survival Rate ; Tumor Suppressor Protein p53 ; metabolism

OBJECTIVETo study the proportion and mechanism of relieving asthma of drug partnership comprising herbal Ephedrae & Pheretima.

METHODTo study relaxant effect on 10 micromol x L(-1) carbachol (CCh) and 10 micromol x L(-1) histamine (His) precontracted isolated tracheal rings and lowering effect on short-circuit current (Isc) increase induced by 10 micromol x L(-1) CCh with 3 proportions of 1:1, 1:3, 1:9 extract.

RESULT1:3 proportions dose-dependently relaxed CCh-precontracted isolated tracheal rings, IC50 of 1:1, 1:3 is 7.5, 15 mg x mL(-1) respectively, 1:9 could not produce 50% inhibition effect on CCh-evoked contraction; 3 proportions also dose-dependently relaxed His-precontracted isolated tracheal rings, IC50 of 1:9, 1:3 and 1:1 is 0.19, 0.61, 1.8 mg x mL(-1) respectively. On the other hand,the orders potency of the decrease effect on CCh-evoked short circuit current increase is 1:3 > 1:1 > 1:9. The difference is not significant (P < 0.05).

CONCLUSIONHerbal Ephedrae & Pheretima had tracheal muscle relaxant and epithelium ion secretion inhibition effect, its mechanism of relieving asthma involved anti-CCh and anti-His effect 1:3 was the most appropriate dosage ratio in the anti-asthmatic drug partnership.

Animals ; Anti-Asthmatic Agents ; administration & dosage ; pharmacology ; Asthma ; physiopathology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Ephedra sinica ; chemistry ; Guinea Pigs ; Histamine Antagonists ; pharmacology ; In Vitro Techniques ; Male ; Materia Medica ; administration & dosage ; isolation & purification ; pharmacology ; Muscle Relaxation ; drug effects ; Muscle, Smooth ; drug effects ; Oligochaeta ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo find out how GATA-1 and GATA-2 behave in the bone marrow of patients with Monge's disease.

METHODSThe levels of mRNA in mononuclear cells (MNC) and proteins of GATA-1 and GATA-2 in the bone marrow of patients with Monge's disease and controls were determined by RT-PCR and immune cytolysis chemical method.

RESULTS(1) All patients and controls expressed GATA-1 mRNA (Monge's disease 1.033 +/- 0.146, Control 0.458 +/- 0.076) and GATA-2 mRNA (Monge's disease 0.451 +/- 0.073, Control 0.185 +/- 0.074). All patients expressed both GATA-1 (positive cell counts 77.3 +/- 33.3, positive score 135.4 +/- 75.4) and GATA-2 ( positive cell counts 29.4 +/- 11.4, positive score 48.4 +/- 19.7). All the controls expressed GATA-1 (positive cell counts 18.1 +/- 11.3, positive score 24.2 +/- 13.4) while 12 of 20 controls expressed GATA-2 ( positive cell counts 5.4 +/- 3.0, positive score 7.3 +/- 4.2). The expression of mRNA and proteins of GATA-1 and GATA-2 in Monge's disease were higher than in controls (P < 0.01). (2) There was a positive correlation between GATA-1 and Hb (P < 0.01), as did between mRNA and proteins of GATA-1 and GATA-2. (3) Both the proteins of GATA-1 and GATA-2 located only in the cytoplasm but not the nucleus.

CONCLUSIONSTwo of inherent genes, GATA-1 and GATA-2 which were expressed at higher levels in patients with Monge's disease than in controls might play significant roles in the pathogenesis of Monge's disease.

Adult ; Altitude Sickness ; metabolism ; GATA1 Transcription Factor ; metabolism ; GATA2 Transcription Factor ; metabolism ; Humans ; Male ; Polycythemia ; metabolism ; RNA, Messenger ; metabolism

OBJECTIVETo prepare microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2 (TIMP-2), and investigate their biological characteristics in vitro.

METHODSChinese hamster ovary (CHO) cells were stably transfected with a human TIMP-2 expression vector, encapsulated in barium alginate microcapsules and cultured in vitro. Morphological appearance of the microcapsules was observed under a light microscope. Cell viability was assessed using MTT (3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide) assay. Enzyme linked immunosorbent assay (ELISA) and reverse zymography were used to confirm the release of biologically active TIMP-2 from the microcapsules. Cryopreservation study of the microencapsulated cells was carried out using dimethyl sulfoxide (DMSO) as preservative agent.

RESULTSThe microcapsules appeared like a sphere with diameter of 300 - approximately 600 microm. The surface of the capsule wall was clearly smooth. The microencapsulated cells survived well and kept proliferating over the 6 weeks observed. No significant difference in TIMP-2 secretion was found between encapsulated and unencapsulated cells. Reverse zymography confirmed the bioactivity of MMP (matrix metalloproteinase) inhibition of TIMP-2. The cryopreservation process did not damage the microcapsule morphology nor the viability of the cells inside.

CONCLUSIONMicroencapsulated engineered CHO cells survive at least 6 weeks after preparation in vitro, and secrete bioactive TIMP-2 freely from the microcapsules.

Animals ; CHO Cells ; Cells, Immobilized ; Cricetinae ; Cryopreservation ; Humans ; Microspheres ; Recombinant Proteins ; biosynthesis ; genetics ; Tissue Engineering ; Tissue Inhibitor of Metalloproteinase-2 ; biosynthesis ; genetics ; Transfection

OBJECTIVETo prevent and manage biliary complications after orthotopic liver transplantation (OLT).

METHODSNinety-five patients of OLT performed at our institute from February, 1999 to December 2002 were retrospectively analysed. Recipient operation was performed using standard method combined with veno-venous bypass in 12 patients and piggyback method in 78 patients and living-related liver transplantation in 5 patients. Biliary reconstruction was performed by end-to-end choledochocholedochostomy (C-C) over a T-tube in 55 patients and without a T-tube in 36 patients while the remaining 4 patients underwent Roux-en-Y choledochojejunostomy (CRY). C-C and CRY were performed by the interrupted or continuous suture with 5 - 0 or 6 - 0 Vicryl or PDS. Routine examination of liver function, Doppler ultrasonography and cholangiography were performed during the follow-up period.

RESULTSBiliary complications occurred in 7 patients (7.3%). Two patients with bile leakage at the anastomotic site developed biliary peritonitis on the seventh and tenth postoperative day and needed reoperation. One patient developed anastomotic biliary stricture one month after the operation and was cured by endoscopic stenting. Two patients developed bile leakage after T-tube removal. One of the two patients was treated conservatively and the other underwent a exploratory laparotomy to ligate the T-tube tract and drain the peritoneal cavity. One patient died of biliary vast syndrome five months after OLT and one patient died of biliary tract necrosis secondary to hepatic artery thrombosis on the tenth postoperative day. One - 42-month (mean 11.4 months) follow-up revealed no biliary stricture in 74 patients. No biliary stone and biliary sludge were detected by Doppler ultrasound and/or cholangiography. Serological examinations proved that liver grafts functioned well in these patients.

CONCLUSIONSTo prevent biliary complications, it is crucial to protect biliary mucosa and arterial blood supply of the common bile duct while harvesting the graft and to obtain perfect mucosa-to-mucosa apposition of no-tension end-to-end anastomosis of the bile duct. Endoscopic dilation and stenting are effective for post-OLT extrahepatic biliary stricture.

Adult ; Aged ; Biliary Tract Diseases ; etiology ; prevention & control ; therapy ; Biliary Tract Surgical Procedures ; adverse effects ; methods ; Female ; Follow-Up Studies ; Humans ; Liver Transplantation ; adverse effects ; methods ; Male ; Middle Aged ; Postoperative Complications ; etiology ; prevention & control ; therapy ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate the chromosomal aberration in sporadic colorectal carcinoma and its association with clinicopathological features.

METHODSComparative genomic hybridization(CGH) was used to screen the changes in the number of DNA sequence copies in 40 sporadic colorectal cancer patients in order to identify regions that contain genes important for the development and progression of colorectal cancer.

RESULTSIn 40 sporadic colorectal cancer, frequent gain at 20 q, 12 q, 13 q, 7 p, 7 q and 16 q were found, while loss was also found at 18 q, 5 q, 4 q, 8 pand 17 p. The number of chromosomal aberration was closely associated with tumor stage(P<0.05). No significant association was found between the number of chromosomal aberration and tumor site, histopathologic type and histologic grade.

CONCLUSIONSChromosomal aberration exists generally in sporadic colorectal carcinoma. The number of chromosomal aberration and gain of 20q are closely associated with tumor stage.

Adult ; Aged ; Aged, 80 and over ; Chromosome Aberrations ; Chromosome Mapping ; Colorectal Neoplasms ; genetics ; pathology ; Comparative Genomic Hybridization ; DNA Probes ; Female ; Gene Dosage ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Oligonucleotide Array Sequence Analysis

OBJECTIVETo induce DNA oxidative damage in colorectal crypt cells by hydrogen peroxide in vitro.

METHODSHydrogen peroxide was diluted into 100, 50, 10, 5 and 1 micromol/L with RPMI 1640. Colorectal crypt cells were treated with peroxide for 10 min, 30 min, 1 h, 1.5 h, 12 h and 24 h respectively. The survival of colorectal crypt cell was measured by MTT method, and the DNA oxidative damage special product, 8-OhdG was detected with immunohistochemical staining. Liner regression was used to measure the time trend of survival rate with SPSS 10.0 software.

RESULTSurvival rate of colorectal crypt cell was 60% and 80% after 10 min of hydrogen peroxide treatment. The longer treatment of hydrogen peroxide, the lower survival rate; the survival rate was reduced to 30% in 24 h. After 10 or 30 min treatment of 100 or 50 micromol/L hydrogen peroxide, the survival rates of colorectal crypt cells were reduced by 20% compared with those of 10, 5 and 1 micromol/L hydrogen peroxide. However, while cells were treated with different concentrations of hydrogen peroxide for 1.0 h or above, there were no differences in cell survival rates. The time trend test results demonstrated that the survival rates of colorectal crypt cells treated with 10, 5 and 1 micromol/L hydrogen peroxide were significantly decreased with the time length of treatment. Colorectal crypt cells treated with different concentrations of hydrogen peroxide for 15 minutes were positively stained brown in cytoplasm and nuclear by immunohistochemistry with 8-OhdG monoclonal antibody.

CONCLUSIONHydrogen peroxide could induce DNA oxidative damage in colorectal crypt cells. And treated with 1 - 10 micromol/L hydrogen peroxide for 10 - 30 min, DNA oxidative damage is apt to be induced in colorectal crypt cell.

Carbazoles ; analysis ; Cells, Cultured ; Colon ; cytology ; drug effects ; metabolism ; Humans ; Hydrogen Peroxide ; Models, Biological ; Oxidative Stress ; drug effects ; Propanolamines ; analysis ; Stem Cells ; cytology ; drug effects