OBJECTIVE: To establish a simple, sensitive in vitro method to evaluate oxygen/glucose deprivation (OGD)-induced injury of brain hippocampal slices in rats. METHODS: Rat hippocampal slices were incubated in 2% 2, 3, 5-triphenyltetrazolium chloride (TTC) solution after oxygen/glucose deprivation. They were then soaked in a measured volume of ethanol and dimethylsulfoxide (50:50) to extract the TTC formazan Product which was then measured by spectrophotometry. OGD induced LDH release was simultaneously measured. RESULTS: Progressive prolongation of OGD induced hippocampal injury resulted in decreased formazan coloration as determined by spectrophotometry. There was a parallel increase in LDH release, thus a negative correlation in these two products was noted. (r=-0.933,P <0.01). The injury was attenuated in the brain slices pre-treated with nimodipine, dexamethasone, and ketamine, but not ONO-1078. CONCLUSION: Solvent extraction and spectrophotometric quantification of formazan represents an objective measurement of OGD-induced injury of rat hippocampal slices.

OBJECTIVETo study the clinical significance of detecting p53 gene mutation expression in colorectal cancer cells of peripheral blood.

METHODSFlow cytometry (FCM) was used to detect p53 gene mutation expression in peripheral blood cancer cells of 128 patients with colorectal cancer. Experimental data were analyzed by SPSS (v.11.0) software.

RESULTSThe lymph node metastasis showed the significant difference statistically (P<0.01) between p53 positive and negative expression in the colorectal cancer patients. The mutation p53 expression associated with existing histological differentiation (r=0.8476, P<0.05). A lymph node metastasis difference was observed between left and right colorectal cancers of mutation p53 positive expression.

CONCLUSIONDetecting the mutation p53 expression in cancer cells of peripheral blood might be helpful to the early diagnosis of colorectal cancer.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; diagnosis ; genetics ; pathology ; DNA, Neoplasm ; analysis ; Female ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Genes, p53 ; genetics ; Humans ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; metabolism ; Tumor Suppressor Protein p53 ; genetics

Objective To determine the prevalence and risk factors of functional constipation (FC) by using Rome Ⅲ criteria in the local adult communities.Methods A stratified randomized and community-based study by multi-stage cluster sampling was employed.A household survey was conducted from April to May 2010.All of the participants were interviewed face-to-face by filling out the self-administered questionnaires which based on Rome Ⅲ criteria for the diagnosis of FC.Self-rating anxiety scale (SAS),self-rating depression scale (SDS) and Athens insomnia scale (AIS) were carried out to evaluate the psychological characteristics and qualities of sleep.Results A total of 7648 subjects fulfilled the questionnaires,with the response rate as 90.0％.211 patients met the Rome Ⅲ criteria,including 90 males and 121 females.The adjusted prevalence rates of FC were 2.5％ in males,3.3％ in females and with an overall rate as 2.9％.The ratio of men to women was 1∶1.32,with significant difference between males and females (P=0.043).The most common group was in the 18-29 year-olds (x2=37.359,P=0.000).FC patients were more likely to be detected in the group with normal BMI (x2=16.087,P=0.002),having received high education (x2=27.604,P=0.000),being intelectuals ( x2=6.922,P=0.031 ) and divorced ( x2=22.000,P=0.000) than in other groups. Multivariate analysis showed that excessive intake of high-fat food was significantly associated with the presence of FC (odds ratio as 1.253,P=0.000),whereas foods with high-fiber (odds ratio as 0.854,P=0.029) might serve as protective factors.Significant differences between FC groups and control groups were found in the incidence of anxiety (with odds ratio as 2.583,P=0.000) and insomnia (odds ratio as 2.443,P=0.000).Conclusion The prevalence of FC in adult communities in Shanghai Songjiang district was not higher than that in other parts of the communities.Excessive intake of high-fat food,anxiety and insomnia might be risk factors for FC and foods with high-fiber contents might serve as protective factors.

OBJECTIVETo study the efficacy, safety and reliability of colonic sac duct for first-stage repair of colorectal anastomotic leakage.

METHODSAn animal model of colon anastomotic leakage was established in 30 Tibet miniature pigs, which were randomly divided into treatment group and control group (n=15). Colon anastomotic leakage in the treatment group was repaired using the colonic sac duct, while the control group received conventional surgical repair. At 7, 14, and 21 days after the surgery, the healing of the anastomotic leakage was evaluated by examining the bursting pressure, tissue microvessel density and hydroxyproline content at the anastomosis.

RESULTSUsing the colonic sac duct, the anastomotic leakage was successfully repaired without death of the pigs or the occurrence of intestinal stenosis or necrosis. At 7 and 14 days after the surgery, the bursting pressure, hydroxyproline contents, and microvessel density in the treatment groups were higher than those in the control group, but such difference was not found at 21 days.

CONCLUSIONColonic sac duct allows effective repair of colon anastomotic leakage, and is especially useful for leakage lasting for 48-72 h complicated by severe abdominal infection.

Anastomosis, Surgical ; adverse effects ; Anastomotic Leak ; etiology ; surgery ; Animals ; Colon ; surgery ; Female ; Male ; Rectum ; surgery ; Swine ; Swine, Miniature

OBJECTIVETo investigate the effects of Xinfeng Capsule (XFC) on pulmonary function and related mechanism in adjuvant-induced arthritis (AA) rats.

METHODSThe rats were randomly divided into five groups: normal control (NC), model control (MC) groups, methotrexate (MTX), tripterygium glycosides tablet (TPT) and Xinfeng capsule (XFC) treatment groups. The adjuvant-induced arthritis model was established by intracutaneous injection of 0.1 mL Freund ' s complete adjuvant in the right paw of rats; the drugs were given 19 d after model establishment. The toe swelling degree (E), arthritis index (AI), pulmonary function, peripheral blood Treg levels, pathological changes of lung tissue and expression of Foxp3, TGF-β1, Smad3, Smad7 proteins in lung tissue were measured 30 d after drug administration.

RESULTSCompared to NC group, the levels of E, AI, alveolitis score, TGF-β1 and Smad3 were significantly increased (P <0.05 or P <0.01); maximum expiratory flow 25% of vital capacity (FEF25),50% maximal expiratory vital capacity flow (FEF50), maximum expiratory flow at 75% of vital capacity (FEF75), maximum mid-expiratory flow (MMF), force peak expiratory flow (PEF), CD4+ CD25+ Treg, Foxp3 and Smad7 were significantly decreased in MC group (P <0.05 or P < 0.01). Compared to MC group,the expression of E, AI, TGF-β1 and Smad3 were reduced, while FEF50, FEF75, MMF, PEF, Treg, Foxp3 and Smad7 were elevated in XFC group (P <0.05 or P <0.01). Compared to XFC group, the level of body mass,FEF25,FEF50, FEF75, MMF and Treg were lower in MTX and TPT groups (P <0.05 or P <0.01).

CONCLUSIONThere are inflamed joints and reduced pulmonary function in rats of adjuvant-induced arthritis. XFC can inhibit paw edema degrees, reduce arthritis response, and improve pulmonary function, which is associated with up-regulating expression of Treg and Foxp3, down-regulating the expression of TGF-β1 and adjusting TGF-β1/Smads signal pathway.

Animals ; Arthritis, Experimental ; drug therapy ; metabolism ; physiopathology ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Forkhead Transcription Factors ; metabolism ; Lung ; drug effects ; pathology ; physiopathology ; Male ; Rats ; Rats, Wistar ; Smad3 Protein ; metabolism ; Smad7 Protein ; metabolism ; T-Lymphocytes, Regulatory ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo observe the injury on micro-skin induced by a self designed micro-skin machine.

METHODSMicro-skin was produced either with the machine or by hand. Cells at the edge of micro-skin were observed by transmission electron microscope. succinic dehydrogenase activity in supernatant of cultivated cells was analyzed, and the cell proliferation of micro-skin was assessed by (3)H-TdR. Twenty patients were enrolled in the study for the observation of the wound healing time between the two groups of micro-skin after being grafted.

RESULTSTransmission electron microscope examination revealed that the cellular injury at the edge of the micro-skin in machine-made group was mild compared with that in man-made group. (3)H-TdR rate was elevated but the activity of succinic dehydrogenase in the supernatant of cultured cells decreased in supernatant of cultured cells of machine produced micro-skin. Wound healing time was shortened in machine made group. (P < 0.05).

CONCLUSIONThe cellular injury at the edge of micro-skin in the machine made group was mild when compared with that in the man-made group with cell proliferation accelerated and wound healing time shortened.

Burns ; surgery ; Cell Division ; Epithelium ; pathology ; Humans ; Microscopy, Electron ; Skin ; ultrastructure ; Skin Transplantation ; methods ; Wound Healing

Objective To investigate the value of endoscopy combined with laparoscopy in the treatment of colorectal polyps and polyp canceration.Methods Different combinations of endoscopic and laparoscopic procedures were employed and the clinical efficacies were compared.Results From January 2004 to September 2006,46 cases with colorectal polyp were treated with endoscopy combined with laparoscopy.Among them,5 cases(10.87%)underwent laparoscopic-assisted endoscopic polypectomy,30(65.22%)endoscopic-assisted laparoscopic resection,6(13.04%)synchronously endoscopic and laparoscopic resection.Five cases were performed further operation after endoscopic polypectomy.According to the pathological findings,21(45.7%)were proved to be polyp canceration,among which 6 were advanced carcinoma,and 3 were found metastasis to the lymph nodes.Among the 41 cases of laparoscopic resection,there was no conversion to an open surgery.Anastomotic leakage was found in 2 cases and anastomotic bleeding in 1.In the 5 cases of laparoscopic-assisted endoscopic polypectomy,no complication was observed.During the period of follow-up(1 to 21 months),no recurrence was detected.Conclusion Endoscopy combined with laparoscopy extends the safety and indications of endoscopic polypectomy,and is minimally invasive to the patients.It is an ideal procedure in the treatment of colorectal polyps and poly carceration.

OBJECTIVETo examine the expression of nestin and neurogenin 3 (Ngn3), the markers of pancreatic stem cells, in the human fetal pancreas.

METHODSThe human fetal pancreas tissue of 12 and 14 weeks were examined for the expression of nestin and Ngn3 using the techniques of immunofluorescence dye and RT-PCR.

RESULTSBoth nestin and Ngn3 expressed widely in 12 and 14 weeks before in human fetal pancreatic tissue. In these positive cells there was no co-expressing insulin or glucagon. There were nestin and Ngn3 co-expressing cells in ducts but not in the islets. The results of RT-PCR also indicated the expression of nestin and Ngn3.

CONCLUSIONSThere was no expression of the markers of mature endocrine cells in the nestin and Ngn3 positive cells, and they were the marks of no-differentiation cells in the human fetal pancreatic tissue.

Basic Helix-Loop-Helix Transcription Factors ; biosynthesis ; genetics ; Fluoroimmunoassay ; Humans ; In Vitro Techniques ; Intermediate Filament Proteins ; biosynthesis ; genetics ; Microscopy, Fluorescence ; Nerve Tissue Proteins ; biosynthesis ; genetics ; Nestin ; Pancreas ; cytology ; embryology ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVESTo evaluate the efficacy of antitumour colon cancer cell vaccine modified by escherichia coli cytosine deaminase (EC-CD).

METHODSMouse colon cancer cell vaccine CT26/CD was established by gene modification using retrovirus plasmid pLCDSN. Its tumorigenicity and effect on liver metastasis model established with wild-type colon cancer were evaluated.

RESULTSCT26/CD was established successfully and proliferated in medium containing 0.6 g/L G418 stably. EC-CD gene expression on these mutant cells was confirmed by RT-PCR. These mutant cells were more sensitive to 5-fluorocytosine (5-FC) compared with the wild-type ones (P=0.000), and presented excellent bystander effect. CT26/CD had the same tumorigenicity as its parental cells (P=0.892). CT26/CD, combined with the prodrug 5-FC, could inhibit tumor progress and live metastasis, and prolonged the survival of the liver metastasis model animals (P=0.000).

CONCLUSIONThe colon cancer cell vaccine modified by EC-CD presented anti-tumor effect in vivo, when treated with the prodrug.

Amino Acid Motifs ; Animals ; Cancer Vaccines ; therapeutic use ; Cell Line, Tumor ; Colonic Neoplasms ; therapy ; Cytosine Deaminase ; genetics ; Escherichia coli ; enzymology ; genetics ; Escherichia coli Proteins ; genetics ; Humans ; Mice ; Mice, Inbred BALB C

BACKGROUND: Studies have confirmed that monosialotetrahexosyl ganglioside can induce human umbilical cord mesenchymal stem cells to differentiate into neuron-like cells, but little is reported on its optimal concentration. OBJECTIVE: To explore the optimal concentration of monosialotetrahexosyl ganglioside that induces human umbilical cord mesenchymal stem cells to differentiate into neuron-like cells in vitro. METHODS: Human umbilical cord mesenchymal stem cells were isolated by using collagenase digestion method, and after expansion, passage 3 cells were randomly allocated into five groups. When 70%-80% of cells were confluent, 50, 100, 150 and 200 mg/L monosialotetrahexosyl ganglioside induction solutions were added in corresponding experimental groups, while cells in the blank control group were cultured in the same volume of L-DMEM medium. Cell morphology was observed under inverted phase contrast microscope. Expression levels of microtubule-associated protein 2, neurofilament protein and glial fibrillary acidic protein were measured by using immunohistochemistry at 6 hours after induction. RESULTS AND CONCLUSION: Human umbilical cord mesenchymal stem cells were isolated successfully and sub-cultured stably. These cells could express surface markers of mesenchymal stem cells. Monosialotetrahexosyl ganglioside at the optimal concentration of 150 mg/L was confirmed to induce the neuron-like differentiation of human umbilical cord mesenchymal stem cells, and differentiated cells could express microtubule-associated protein 2 and neurofilament protein as neuron-specific markers.