BACKGROUND: Because of complicated physiological environment and difficulty to control experimental conditions, it is difficult to get satisfactory results from in vivo studies of cell mechanics.OBJECTIVE: To study the action and mechanism of p38MAPK signaling pathways on myoblast apoptosis based on successful construction of in vitro mechanical stimulation models.METHODS: The C2C12 cells cultured in vitro were divided into control group and SB203580 treatment group. Cyclic tensile stress was applied on the C2C12 myoblast cells for 0, 6, 12 and 24 hours in each group. The Flexcell Strain Unit-5000T was used to expose C2C12 myoblast cell to an equiaxial cyclic of 15% magnitude and a frequency of 10 cycles/min, each cycle including the 3 s stretch and 3 s relaxation. Hoechst 33258 fluorescent staining and optical microscope were used to detect cell apoptosis. RT-PCR, flow cytometric analysis were used to observe the apoptosis of C2C12 myoblast cells and Western blotting were used to detect the activity of p38MAPK and p-p38MAPK. RESULTS AND CONCLUSION: The optical microscope tested the change in the morphology. Hoechst 33258 staining showed that after treatment with cyclic stress, the cell took the typical appearance of apoptosis with chromatin condensation and apoptotic bodies. RT-PCR and flow cytometry showed that with the extension of time the rate of the apoptosis of C2C12 myoblast cell increased. And cells imposed SB203580 before imposing cyclical tensile stress, the results showed that the apoptosis was markedly affected, and the p-p38MAPK expression declined apparently. These findings demonstrate that p38MAPK signaling pathways in stress mediated into C2C12 myoblast cell apoptosis plays an important role.

OBJECTIVETo study reverse effect of the oxidative damage on cartilage cells of velvet antler polypeptides (VAPS), and to investigate the main mechanism of VAPS to protect cartilage cells through antioxidant.

METHODSFifteen Japanese white rabbits of 5-month-old were selected in this study. Animal model was established by method of Hulth osteoarthritis animal model. The anterior and posterior cruciate ligament and medial collateral ligament were cut off and medial meniscus were cut, articular cartilage cell cultured in vitro. Cells in the sham operation group was the normal control group, osteoarthritis cartilage cells in the model groups were added VAPS 6.25, 12.5, 25 microg/ml respectively. A group of animals were sacrificed every week form the ninth weeks(two months) and the cartilage cells were isolated and cultured. For 8 weeks,the reactive oxygen species level in chondrocytes were detected by DCFH-DA, the content of NO, SOD and GSH-Px in cell culture supernatant were detected by Griess method.

RESULTSDCFH-DA detection of intracellular reactive oxygen species was (5.46 +/- 0.46)in the control group, (12.08 +/- 0.74) in the model groups. The model group compared with the control group by t test with the P value less than < 0.001. DCFH-DA detection of intracellular reactive oxygen species was (9.81 +/- 0.59)in VAPS 6.25 microg/ ml group, (7.83 +/- 0.63) in the VAPS 12.5 microg/ml group, (6.89 +/- 0.71) in the VAPS 25 microg/ml group, as compared with model group there were statistically significant difference (P < 0.05). The content of NaNO2, SOD and GSH-Px in osteoarthritis model group was (5.60 +/- 0.45) microM, (38.56 +/- 12.53) U/ml and (151.90 +/- 25.60) U, as compared with control group there were statistically significant difference (P < 0.001, P < 0.05); The content of NaNO2 was (4.34 +/- 0.39), M in VAPS 6.25 microg/ml group, (3.67 +/- 0.36) microM in the VAPS 12.5 microg/ml group, (3.20 +/- 0.27) microM in the VAPS 25 microg/ml group, as compared with model group there were statistically significant difference (P < 0.01). The content of SOD was (49.91 +/- 5.77) U/ml in VAPS 6.25 microg/ml group, (54.05 +/- 5.27) U/ml in the VAPS 12.5 microg/ml group, (57.44 +/- 5.70) U/ml in the VAPS 25 microg/mL group, as compared with model group there was statistically significant (P < 0.05). The content of GSH-Px was (172.50 +/- 18.65) U in VAPS 6.25 microg/ml group, (202.10 +/- 21.60) U in the VAPS 12.5 microg/ml group, (315.80 +/- 10.50) U in the VAPS 25 microg/ml group, the VAPS 12.5 microg/mL group and VAPS 25 microg/ml group was compared with model group, there were statistically significant difference (P < 0.01).

CONCLUSIONThe VAPS have antioxidative damage effect of osteoarthritis cartilage cells within a certain range and dose-dependent manner. It may be the main mechanism for velvet antler polypeptides to treat osteoarthritis.

Animals ; Antlers ; chemistry ; Cartilage ; drug effects ; metabolism ; pathology ; Female ; Glutathione ; blood ; Male ; Nitric Oxide ; blood ; Osteoarthritis ; blood ; metabolism ; pathology ; Oxidative Stress ; drug effects ; Peptides ; pharmacology ; Rabbits ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; blood

Objective To detect the argyrophilic proteins in nucleolar organizer regions(Ag-NORs) that express rDNA and rRNA proliferation of T lymphocytes before chemotherapy and after complete remission(CR) in children with primary acute leukemia(AL).Methods The argyrophilic granules area of NOR/nuclear area(I.S%) of T lymphocytes was detected by image analysis system in peripheral blood of 42 patients before chemotherapy and after CR and 30 normal children.Results I.S% in the patients before chemotherapy(5.06%?1.36%) were significantly lower than those in the healthy donors(7.51%?1.06%)(t=8.238 P0.05).Conclusion These results suggest that decrease of Ag-NORs expresses the evidence for tumour induced suppression of immune function of T cells in children with AL prior to treatment.

Humans ; Deglutition ; Time Factors ; Language

OBJECTIVETo explore the effect of Changji'an Capsule (CA) on mRNA expressions of neuropeptide Y (NPY) in the hypothalamus and colon and serum levels of adreno-cortico-tropic hormone (ACTH) in rats of diarrhea predominant irritable bowel syndrome (IBS-D) model rats.

METHODSTotally 48 SD rats were randomly divided into six groups, i.e., the normal control group, the model group, the Pinaverium Bromide group (PB, 0.018 g/kg), the high dose CA group (2.812 g/kg), the medium dose CA group (1.406 g/kg), and the low dose CA group (0.703 g/kg), 8 in each group. The IBS-D rat model was established by using separation of breast milk + stimulation of acetic acid + constraint of four limbs. Normal saline was given to rats in the normal control group and the model group. All medication lasted for 14 successive days by gastrogavage. The serum content of ACTH was detected by enzyme linked immunosorbent assay (ELISA). The expressions of NPY mRNA in the colon and the hypothalamus were detected using real-time fluorescence quantitative PCR.

RESULTSCompared with the normal control group, the serum ACTH content significantly increased (P < 0.01), the NPY mRNA expression in the colon and the hypothalamus obviously decreased (P < 0.01) in the model control group. Compared with the model group, the serum ACTH obviously decreased in the high dose CA group, the medium dose CA group, and the PB group (P < 0.01, P < 0.05). The NPY mRNA expression in the colon and the hypothalamus were obviously up-regulated in the high dose CA group, the medium dose CA group, the low dose CA group, and the PB group (P < 0.05).

CONCLUSIONSCA could modulate the abnormity of brain-gut axis of IBS-D rats possibly by up-regulating NPY mRNA expressions in the hypothalamus and the colon and down-regulating the ACTH content in the hypothalamic-pituitary-adrenal axis.

Adrenocorticotropic Hormone ; blood ; Animals ; Colon ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hypothalamus ; metabolism ; Irritable Bowel Syndrome ; metabolism ; Male ; Neuropeptide Y ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the effect of occupational stress on ambulatory blood pressure.

METHODS30 male healthy workers from the refrigerator assembly line in Henan province in China were investigated. Psychosocial work conditions were measured by using the Job Demand-control Model, the Effort-reward Imbalance Model questionnaires and Occupational Stress Measurement Scale. Ambulatory blood pressure(ABP) was measured by using mobile ABP monitor. The t test was utilized to analyze the difference of parameters of ABP monitoring between different groups of occupational stress and other variables scores. The stepwise regression analysis was used to analyse the effect of occupational stress factors on parameters of ABP.

RESULTS(1) As to stressors, systolic blood pressure variability (SBPV), mean arterial blood pressure variability (MABPV) and heart rate at 30 minute after work in workers with high role conflict score were significantly higher than those in workers with low score (P < 0.05). Workers with high skill utilization score had significantly lower mean systolic blood pressure (SBP) at 30 minute after work than workers with low score (P < 0.05). Diastolic blood pressure variability (DBPV) and heart rate variability (HRV) in workers with high decision latitude score were significantly higher than those in workers with low score (P < 0.05). Workers with high job psychological demands score had significantly higher SBPV, DBPV and MABPV than workers with low score (P < 0.05). Heart rate-pressure product(RPP) and SBPV in workers with high effort score were significantly higher than those in workers with low score (P < 0.05). Workers with low rewards score had higher mean heart rate and heart rate at 30 minute after work than workers with high score (P < 0.05). (2) For personalities, workers with high work locus of control score had significantly higher mean diastolic blood pressure (DBP) and mean arterial blood pressure (MABP) than workers with low score (P < 0.05). Workers with high patience score had significantly lower mean SBP at 30 minute after work than workers with low score (P < 0.05). Heart rate at 30 minute after work in workers with high organization commitment score was significantly lower than that in workers with low score (P < 0.05). (3) Concerning buffer factors, HRV in workers with high control strategies score were significantly lower than that in workers with low score (P < 0.05). Workers with low supervisor support score had higher RPP and MABPV than workers with high score (P < 0.05). (4) In the multiple stepwise regression, daily life stress affected SBPV (R2 = 0.12) and MABPV (R2 = 0.05), depression was related to DBPV at 30 minute after work (R2 = 0.15) and SBPV (R = 0.03), mental health was predictor of MABPV (R2 = 0.07) and negative affection was predictor of heart rate at 30 minute after work (R2 = 0.24).

CONCLUSIONSOccupational stressors, personality and social support have effect on parameters of ABP. Parameters of ABP monitoring could be used to evaluate occupational stress in the field research.

Adult ; Blood Pressure ; physiology ; Blood Pressure Monitoring, Ambulatory ; Burnout, Professional ; physiopathology ; Humans ; Male ; Multivariate Analysis ; Regression Analysis ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo explore the effect of occupational stress on hypertension.

METHODS498 workers whose accumulative length of service was more than two years were investigated with questionnaire by method of cluster sampling from a thermal power plant in Henan province in China; 446 respondents returned qualified questionnaire including 281 male and 165 female Han People. After the patients with secondary hypertension, diabetes patients, and patients with liver or kidney disease were excluded, 84 workers (58 males and 26 females) were diagnosed as hypertension. The occupational stressors, personalities, buffering factors and occupational strain were measured by using the Job Demand-control Model, the Effort-reward Imbalance Model questionnaires and Occupational Stress Measurement Scale. Main risk factors for the development of hypertension such as heredity, body mass index, high salt diet, alcohol use, smoking habit and lack of physical activity were investigated. 498 whole blood samples were collected from workers in field epidemiologic survey. All of the samples were detected TG, CHO and FPG by common biochemistry methods. Multivariate logistic regression analysis were used to determine the relationship between occupational stressors and prevalence rate of hypertension. The difference of morbidity of hypertension between different stress level subjects was analyzed by chi2 test.

RESULTS(1) Logistic regression analysis of the hypertension by all occupational stressors and risk factors of hypertension indicated that not only some common factors such as parents' hypertensive history, BMI, alcohol use and TG, but also responsibility for person, work locus of control and social support were significantly correlated with elevated risks of hypertension. (2) Logistic regression analysis of the hypertension by main dimensions of effort-reward imbalance model and risk factors of hypertension indicated that parents' hypertensive history, BMI, alcohol use, TG, and effort were significantly correlated with elevated risks of hypertension. Logistic regression analysis indicated the risk of hypertension had an effect on the FRI and effort (OR was 1.71 and 2.43 respectively). (3) For the job strain model, results indicated that parents' hypertensive history, UMI, alcohol use, TG, work locus of control and social support were significantly correlated with elevated risks of hypertension. But the main dimensions of job strain model (job demands and decision latitude) didn't enter regression equation. (4) The difference of prevalence of hypertension between high- and low stress level groups in male was statistically significant (OR = 3.13, P < 0.01), but the case was not the same in female (P > 0.05).

CONCLUSIONSOccupational stress might be risk factor of hypertension; The predictive power of effort-reward imbalance model for the development of hypertension would be larger than that of job strain model.

Adult ; Burnout, Professional ; complications ; Chi-Square Distribution ; China ; Cross-Sectional Studies ; Female ; Humans ; Hypertension ; etiology ; Logistic Models ; Male ; Middle Aged ; Risk Factors ; Sampling Studies ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo investigate the role of the expression of Ezh2, Runx3 and caspase-3 proteins and their correlation in the pathogenesis of endometrial carcinoma.

METHODSExpression of Ezh2, Runx3 and caspase-3 proteins was examined by tissue microarray technique and immunohistochemistry (SP method) in 72 cases of endometrial adenocarcinomas, 60 endometrial hyperplasia and 30 normal endometrial tissues.

RESULTSThe positive expression rates of Ezh2, Runx3 and caspase-3 proteins in endometrial adenocarcinomas were 83.3% (60/72), 26.4% (19/72) and 33.3% (24/72), respectively. The positive rate of Ezh2 protein in endometrial carcinomas was higher than that in normal endometrium and endometrial hyperplasia (16.7%, 33.3%, 63.3%;P < 0.05). However, the positive rate of Runx3 in endometrial carcinomas was lower than that in normal endometrium and endometrial hyperplasia (80.0%, 56.7%; P < 0.01). The positive rate of caspase-3 protein in endometrial carcinomas was lower than that in normal endometrium and endometrial hyperplasia (86.7%, 73.3%, 63.3%; P < 0.01). Positive expression of Ezh2 and Runx3 was related to the histological grade, FIGO stage, and depth of invasion of endometrial adenocarcinomas (P < 0.05), but it was not related to the lymph node metastasis (P > 0.05). Positive expression of caspase-3 protein was related to the histological grade (P < 0.05), but it was not related to the FIGO stage, depth of invasion and the lymph node metastasis of endometrial adenocarcinomas (P > 0.05). The expression of Ezh2 protein was negatively correlated to that of Runx3 (r(s) = -0.262, P < 0.05).

CONCLUSIONSAbnormal expression of Ezh2, Runx3 and caspase-3 proteins is associated with the development and progression of endometrioid adenocarcinoma. Combined analysis of Ezh2, Runx3 and caspase-3 may offer prognostic information for patients with endometrial cancer.

Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Caspase 3 ; metabolism ; Core Binding Factor Alpha 3 Subunit ; metabolism ; DNA-Binding Proteins ; metabolism ; Endometrial Hyperplasia ; metabolism ; pathology ; Endometrial Neoplasms ; metabolism ; pathology ; Endometrium ; metabolism ; Enhancer of Zeste Homolog 2 Protein ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Grading ; Neoplasm Invasiveness ; Neoplasm Staging ; Polycomb Repressive Complex 2 ; Transcription Factors ; metabolism

OBJECTIVETo compare the therapeutic effects between acupuncture based on syndrome differentiation and analgesic on abdominal postoperative pain.

METHODSOne hundred cases of abdominal postoperative pain were randomly divided into two groups, 50 cases in each one. In acupuncture group, the treatment was applied according to meridian differentiation and point selection on the affected meridian. Ashi points near to the incision as the main points and those closely connected with Zangfu functions were selected, such as Yanglingquan (GB 34), Taichong (LR 3) and Zusanli (ST 36), etc. In medication group, muscular injection of Bucinnazine was administered. The severity of pain was evaluated with Visual Analogue Scale (VAS) before and after treatment.

RESULTSThe remarkably effective rate in acupuncture group was 60.0% (30/50), which was markedly better than that 28.0% (14/50) in medication group (P < 0.01). VAS scores in 30 min and 4 h after treatment as well as 24 h after operation in two groups were all reduced remarkably as compared with those before treatment (all P < 0.01), indicating the satisfactory analgesia in treatment. VAS scores in acupuncture group were lower apparently than those in medication group in 30 min and 4 h after treatment (P < 0.01,P < 0.05).

CONCLUSIONAcupuncture has quick analgesia in treatment of abdominal postoperative pain, which is superior to muscular injection of Bucinnazine because of its advantages of long-term and significant efficacy.

Abdomen ; surgery ; Acupuncture Analgesia ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Pain Measurement ; Pain, Postoperative ; therapy ; Young Adult

AIMTo establish a sensitive and efficient reporter gene-based screening model for finding agonists of estrogen receptor beta subtype.

METHODSA recombinant vector pTAL-ERE-SEAP was constructed by inserting a synthetic sequence composed of five estrogen responsive elements in front of promoter of pTAL-SEAP vector. pTAL-ERE-SEAP was then transfected into human embryonic kidney (HEK293) cells. G418 (200 microg x mL(-1)) was added to select positive clones that can be induced by E2 to express reporter gene SEAP. The speciality was tested by several ligands of relative nuclear receptors of the same family. The stability of the model, the time-effect relationship, the dose-response relationship, and the immunocytochemistry staining of ERbeta expression after transfection were observed. 2 622 compounds were screened by using this model.

RESULTSStably transfected clones were obtained. The expression levels of reporter gene SEAP of positive clones was induced by E2 in a dose-response and time-effect relationship manners. The Z' factor value was 0.7. The expression levels of dexamethasone and other ligands were low. The result of immunocytochemistry staining showed the expression of ERbeta. E2 had no proliferating effects on stably transfected clones.

CONCLUSIONStably transfected positive clones transfected with recombinant vector pTAL-ERE-SEAP were obtained. The positive clones may be used to screen for agonists of estrogen receptor beta subtype by measurement of luminescent value of expressed SEAP in wells of microlitre plate.

Alkaline Phosphatase ; genetics ; metabolism ; Cell Line ; Estradiol ; pharmacology ; Estrogen Receptor beta ; agonists ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; Genetic Vectors ; Humans ; Immunohistochemistry ; Promoter Regions, Genetic ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Response Elements ; genetics ; Transfection