ObjectiveTo study the relationship between the antitumor immunoreaction and the cell apoptosis in esophageal carcinoma.MethodsThe IL-1β converting enzyme, Fas and FasL protein were labeled by LSAB immunohistochemistric method in 46 cases of esophageal carcinoma, of which 13 cases were labeled by TdT mediated dUTP nick end labeling (TUNEL).ResultsThe positive rates of ICE, Fas and FasL proteins in the cancer nests were 78.26%, 60.87% and 47.83%. The expression of ICE protein was related to the histological grading of the cancer.Conclusions The expression of Fas, FasL protein may be related to the immunological escape of the cancer cell; the expression of ICE was related to the histological grading of the cancer.

Cold Temperature ; Dinoprost ; analogs & derivatives ; urine ; Dust ; Humans ; Interleukin-6 ; urine ; Thromboxane B2 ; analogs & derivatives ; urine ; Weather

OBJECTIVETo develop a double antibody sandwich ELISA assay for quantitative determination of recombinant human interferon alpha1b.

METHODSMouse monoclonal antibodies with different binding site on rIFN-alpha1b were screened to select optimized candidates as coating and HRP-labeled index antibodies respectively. And a double antibodies sandwich ELISA was assembled; the reliable lower detection limit, specificity, accuracy and reproducibility were evaluated and validated.

RESULTSThe quantitative sandwich ELISA had a reliable lower detection limit of 10 ng/ml, with a liner detection range 10-100 ng/ml (R2 = 0.992), variation coefficient inter-plates is less than 10%.

CONCLUSIONThe developed sandwich ELISA was a sensitive and specific, accuracy and reproducibility method for quantitative determination of recombinant human interferon alpha1b in final product.

Animals ; Antibodies, Monoclonal ; analysis ; Enzyme-Linked Immunosorbent Assay ; instrumentation ; methods ; Humans ; Interferon-alpha ; blood ; Mice ; Mice, Inbred BALB C

The present study was to investigate the effect of Salvia miltiorrhiza Bunge. f. alba (SMA) pharmacological pretreatment on apoptosis of cultured hippocampal neurons from neonate rats under oxygen-glucose deprivation (OGD). Cultured hippocampal neurons were randomly divided into five groups (n = 6): normal plasma group, low dose SMA plasma (2.5%) group, middle dose SMA plasma (5%) group, high dose SMA plasma (10%) group and control group. The hippocampal neurons were cultured and treated with plasma from adult Wistar rats intragastrically administered with saline or aqueous extract of SMA. The apoptosis of neurons was induced by glucose-free Earle's solution containing 1 mmol/L Na2S2O4 and labeled by MTT and Annexin V/PI double staining. Moreover, protein expressions of Bcl-2 and Bax were detected by immunofluorescence. The results showed that few apoptotic cells were observed in control group, whereas the number of apoptotic cells was greatly increased in normal plasma group and low dose SMA plasma group. Both middle and high dose SMA plasma could protect cultured hippocampal neurons from apoptosis induced by OGD (P < 0.05). The protective effect of high dose SMA plasma was stronger than that of middle one (P < 0.05). Compared to control, normal plasma and low dose SMA plasma groups, middle and high dose SMA plasma groups both showed significantly higher levels of Bcl-2 (P < 0.05 or 0.01), whereas expressions of Bax was opposite. There were no significant differences of Bcl-2 and Bax expressions between middle and high dose SMA plasma groups. Number of Bcl-2- and Bax-positive cells had similar tendency. Bcl-2/Bax (number of positive cells) ratio was higher in high dose SMA plasma group than those of all the other groups (P < 0.05 or 0.01). These results suggest that pharmacological pretreatment of blood plasma containing middle and high dose SMA could raise viability and inhibit apoptosis of OGD-injured hippocampal neurons by up-regulating the expression of Bcl-2 and down-regulating the expression of Bax.
Animals ; Apoptosis ; drug effects ; Cell Hypoxia ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Female ; Glucose ; metabolism ; Hippocampus ; cytology ; Ischemic Preconditioning ; methods ; Male ; Neurons ; cytology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Wistar ; Reperfusion Injury ; prevention & control ; Salvia miltiorrhiza ; chemistry ; bcl-2-Associated X Protein ; metabolism

The present study was to investigate the effect of cerebrospinal fluid (CSF) from the rats with hypoxic preconditioning (HPC) on apoptosis of cultured hippocampal neurons in neonate rats under oxygen glucose deprivation (OGD). Adult Wistar rats were exposed to 3 h of hypoxia for HPC, and then their CSF was taken out. Cultured hippocampal neurons from the neonate rats were randomly divided into four groups (n = 6): normal control group, OGD group, normal CSF group and HPC CSF group. OGD group received 1.5 h of incubation in glucose-free Earle's solution containing 1 mmol/L Na2S2O4, and normal and HPC CSF groups were subjected to 1 d of corresponding CSF treatments followed by 1.5 h OGD. The apoptosis of neurons was analyzed by confocal laser scanning microscope and flow cytometry using Annexin V/PI double staining. Moreover, protein expressions of Bcl-2 and Bax were detected by immunofluorescence. The results showed that few apoptotic cells were observed in normal control group, whereas the number of apoptotic cells was greatly increased in OGD group. Both normal and HPC CSF could decrease the apoptosis of cultured hippocampal neurons injured by OGD (P < 0.01). Notably, the protective effect of HPC CSF was stronger than that of normal one (P < 0.01). Compared to OGD group, normal and HPC CSF groups both showed significantly higher levels of Bcl-2 (P < 0.01), and Bcl-2 expression level in HPC CSF group was even higher than that in normal CSF group (P < 0.01). Whereas the expressions of Bax in normal and HPC CSF groups were significantly lower than that in OGD group (P < 0.01), and the Bax expression in HPC CSF group was even lower than that in normal CSF group (P < 0.01). These results suggest that CSF from hypoxic-preconditioned rats could degrade apoptotic rate of OGD-injured hippocampal neurons by up-regulating expression of Bcl-2 and down-regulating expression of Bax.
Animals ; Animals, Newborn ; Apoptosis ; physiology ; Cell Hypoxia ; physiology ; Cells, Cultured ; Cerebrospinal Fluid ; physiology ; Female ; Glucose ; metabolism ; Hippocampus ; cytology ; pathology ; Hypoxia ; cerebrospinal fluid ; physiopathology ; Ischemic Preconditioning ; Male ; Neurons ; pathology ; Oxygen ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Wistar ; bcl-2-Associated X Protein ; metabolism

OBJECTIVEIn order to provide theoretic evidence of exercise preventing insulin resistance, we studied the effects of high-fat diet and aerobic exercise on expression of mammalian target of rapamycin/ribosomal protein kinase (mTOR/S6K1) in skeletal muscle of insulin resistant mice.

METHODS8-week old, C57BL/6 mice were divided into two groups: which are normal chow and high-fat diet groups, each group has 20 animals feeding by normal chow and high fat diet respectively. 8-weeks later, mice from high fat diet group were proved to have insulin resistance symptoms and after that time point the mice were regrouped. The normal chow group was randomly divided into normal chow diet control group (NC) and normal chow exercise group (NE), and the high fat diet group was randomly divided into high fat diet control group (HC) and high fat diet exercise group (HE). All exercise groups were required to running on treadmill for 6 weeks. Mice were acclimatized to the motorized treadmill by running 60 min per day at the intensity of 75% VO2max five days per week for 6 weeks. By the end of training, we observed the changes of glucose tolerance by oral glucose tolerance test and morphology of pancreatic islet under microscope. Insulin concentration was measured by ELISA. Northern blot and Western blot were performed to detect mTOR, S6K1 (and/or pS6K1-Thr389) mRNA and protein expression in skeletal muscle.

RESULTSBy comparing with NC group, the effect of high-fat diet increased the body weight, fasting serum insulin level and the area of pancreatic islet of the mice in HC group significantly. Furthermore, glucose tolerance was impaired. After 6-week aerobic exercise, above value of descriptive index were significantly decreased in skeletal muscle in HE group. In addition, OGTT was also improved. The expression of mTOR, S6K1 (and/or pS6K1-Thr389) mRNA and protein were significantly decreased.

CONCLUSIONmTOR/S6K1 signaling pathway plays an important role in the development of diet induced insulin resistance. Aerobic exercise attenuates insulin resistance by increasing skeletal muscle insulin sensitivity evidenced by increased energy metabolism decreased activity of mTOR/S6K1 signaling pathway.

Animals ; Body Weight ; Diet, High-Fat ; Glucose Tolerance Test ; Insulin ; blood ; Insulin Resistance ; Male ; Mice ; Mice, Inbred C57BL ; Muscle, Skeletal ; metabolism ; Physical Conditioning, Animal ; physiology ; Ribosomal Protein S6 Kinases, 90-kDa ; metabolism ; Signal Transduction ; TOR Serine-Threonine Kinases ; metabolism

OBJECTIVETo discuss effective operative method for posterior edge separation of lumbar vertebral body.

METHODSTwelve cases of posterior edge separation of lumbar vertebral body were categorized as lateral type and central type, and were operated upon with limited-incision using lamina retractor. All patients were male and with an average age of 30 years (range, 23 to 40 years). The average history of disease was 17.9 months. In lateral type cases, separation bone was in the posterolateral lower edge of L4 vertebral body in 1 case, in the posterolateral upper edge of L5 vertebral body in 3 cases and in the posterolateral upper border of S1 vertebral body in 6 cases, all with ipsilateral disc herniation. Lateral type cases were treated with unilateral fenestration and removal of disc nucleus and posterior bony edge. Separation bone was in the central upper edge of S1 vertebral body with central disc herniation in 2 central type cases. Two central type cases were treated with bilateral fenestration and removal of disc nucleus and posterior bony edge.

RESULTSAll patients were followed up for 12 to 36 months, and the average follow-up was 22.5 months. According to Macnab postoperative evaluation criterion, the results were excellent in 10 cases, good in 2 cases.

CONCLUSIONLimited-incision for posterior edge separation of lumbar vertebral body has characteristic of less trauma and convenience. It is an effective method for removal posterior bony edge.

Adult ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; diagnostic imaging ; pathology ; surgery ; Male ; Postoperative Period ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

Objective To describe the prevalence and risk factors for suicide ideation among college students in Chongqing city. Methods Data on suicide ideation and related factors were collected from 9808 college students at 11 colleges randomly selected in Chongqing. A multiple logistic regression model was used to identify risk factors for suicide ideation. Results 1279 (13.0%) of the 9808 students reported suicide ideation and the constituent ratio of boys and girls was 3:4 whileriskfactors for suicide ideation were ranked as follows: high frequency of feeling hopeless in prior year (OR=5.07,95%CI: 4.27-6.02); having psychological problems in recent 1 month that affecting daily lives and learning(2.07,1.79-2.38); relatives having suicide behavior (1.77,1.52-2.08); having had sexual experience (1.95,1.65-2.30); being female (1.66,1.45-1.90) and friends who had had suicide attempts(1.46,1.28-1.67);having diseases in the last 1 month that affecting daily lives and learning (1.29,1.08-1.52). Conclusion The prevalence of suicide ideation among these college students was high that calls the development,implementation and assessment of suicide prevention plans for college students that focusing on the risk factors identified for suicide ideation.

OBJECTIVEThe growth and metastasis of solid tumors are dependent on angiogenesis. Endostatin, the C-terminal proteolytic fragment of collagen XVIII, is a potent endogenous angiogenesis inhibitor. The authors designed a topical antiangiogenic gene therapy with recombinant human endostatin adenovirus (Ad-hEndo) and assessed its effects on the inhibition of angiogenesis in vitro, and tumor growth and metastasis in vivo.

METHODSMalignant cells (A549) were infected with Ad-hEndo. The expression of recombinant protein and the inhibition of cultured human umbilical vein endothelial were investigated. Immunodeficient A549 nude mice were treated with intratumoral injection of Ad-hEndo, the empty vector Ad-control or saline (NS). The dose-response, side effects, and serum concentration of endostatin were observed.

RESULTSRecombinant endostatin protein was detected in the infected tumor cells with different MOI Ad-hEndo and its inhibitory effect on endothelial cells growth was shown. In animal study, the volume of tumor and the number of pulmonary metastatic lesions in the Ad-hEndo treatment group were significantly smaller than those in the control groups (P<0.05).

CONCLUSIONThe present findings provide evidence of the anti-tumor effects of the endostatin and may be important for the further use of it in topical antiangiogenic gene therapy of cancer.

Adenoviridae ; genetics ; Animals ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cells, Cultured ; Disease Models, Animal ; Endostatins ; biosynthesis ; genetics ; therapeutic use ; Endothelial Cells ; Genetic Therapy ; Genetic Vectors ; Humans ; Kidney ; cytology ; metabolism ; Mice ; Mice, Nude ; Neoplasm Metastasis ; Neoplasm Transplantation ; Neoplasms, Experimental ; prevention & control ; Random Allocation ; Recombinant Proteins ; biosynthesis ; genetics ; therapeutic use ; Transfection ; Umbilical Veins ; cytology ; Xenograft Model Antitumor Assays

Objectives To screen the high-risk population of stroke in China using color Doppler flow imaging (CDFI)and to establish a stroke risk prediction model in Chinese population in order to prevent and treat stroke early. Methods Forty-one base hospitals and 715 286 people in the project areas of the first 6 provinces of China conducted routine physical examinations and investigated the related risk factors for cardiocerebrovascular diseases from July 2011 to April 2012 using a cross-sectional study,among them 61 860 patients underwent carotid CDFI screening,and 49 386 of them were high-risk population (exposed to≥3 risk factors). The bilateral common carotid interma-media thickness (IMT),the number of plaques and the degree of carotid stenosis were screened and documented. And whether carotid IMT thickening or not,with or without carotid plaques,and degree of carotid artery stenotic rate 0-49% and≥50% were performed by multivariate logistic regression analysis with the risk factors for stroke,respectively. Results (1)Logistic regression analysis showed that hypertension,atrial fibrillation,smoking,and lack of physical exercise were the independent risk factors for carotid IMT thickening (hypertension:OR,1. 17;95%CI 1. 12-1. 22;atrial fibrillation:OR,1. 15;95%CI 1. 09-1. 21;smoking:OR,1. 13;95%CI 1. 08-1. 17;and lack of physical exercise:OR,1. 12;95%CI 1.08-1. 16). (2)Hypertension,atrial fibrillation, smoking,and diabetes were the independent risk factors for carotid plaque and carotid artery stenosis rate≥50%(carotid plaque,hypertension:OR,1. 55;95%CI 1. 47-1. 62;atrial fibrillation:OR,1. 13;95%CI 1.06-1. 21;smoking:OR,1. 16;95%CI 1. 11-1. 22;and diabetes:OR,1. 30;95%CI 1. 24-1. 37). Carotid stenosis rate≥50%,hypertension:OR,1. 78;95%CI 1.55-2. 03;atrial fibrillation:OR,1. 59;95%CI 1. 39-1. 81;smoking:OR,1. 33;95%CI 1. 20-1. 48;and diabetes:OR,1. 30;95%CI 1. 17-1. 45. Simple obesity did not increase the incidences of carotid atherosclerotic plaque and carotid artery stenosis ≥50%(OR,0. 78, 0.83;95%CI 0. 75-0. 82 ,0. 75-0. 92,respectively). Conclusions Neck vascular ultrasound can be used as a valuable means for screening high-risk population and detecting risk factors of stroke. It has an important clinical significance for the early diagnosis and treatment of carotid atherosclerosis disease.