Objective To investigate the effects of different magnitudes of mechanical strain on proliferation and alkaline phosphatase (ALP) activity in mouse osteoblast-like cells (MC3T3-E1). Methods MC3T3-E1 cells were subjected to 0%, 6%, 12%, 18% elongation for 24h and 48h by using multi-passage cell stress loading system respectively. MTT colorimetric method was used to assess the proliferation of the cell, ALP activity was detected by ALP assay kit. Results The proliferation of MC3T3-E1 cells was increased significantly 24h and 48h after mechanical strain treatment concomitant with increasing stretching force (P

The changes of CD4(+)CD25(+) regulatory T cells (CD4(+)CD25(+) Treg) and Foxp3 mRNA in peripheral blood mononuclear cells (PBMCs) from patients with asthma were investigated in order to elucidate the possible roles of CD4(+)CD25(+) Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls (normal control group) and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4(+)CD25(+) Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4(+)CD25(+) Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control group (P<0.05). Although the CD4(+)CD25(+) Treg ratio and Foxp3 mRNA of remission group also lower than that of normal control group, there was no significant difference between them (P>0.05). As compared with persistent group, exacerbation group had lower CD4(+)CD25(+) Treg ratio and Foxp3 mRNA (P<0.05). It was indicated that the decrease of CD4(+)CD25(+) Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.

Objective:To analyze the aberrant methylation of p16 gene in sera from primary liver cancer patients and to evaluate the clinical significance. Methods:A methylation-specific PCR was performed for the detection of promoter hypermethylation of p16 gene in blood DNA from 64 cases of PLC patients,and to anamyze the relationship between the aberrant methylation of p16 gene and the clinical pathological data . Results:76.6%(49/64)of the sera from 64 cases of PLC patients showed hypermethylation for p16 promoter,whereas no methylated p16 gene promoter were found in sera from liver benign diseases patients and normal control. Methylated p16 gene promoter in sera did not strongly correlated with HBsAg, AFP,stage ,metastasis and differentiation in PLC. Conclusion:Detection of the aberrant methylation of p16 gene in blood DNA from PLC patients might offer an effective means for the earlier auxillary diagnosis of the malignancy.

Objective: To analyze the aberrant methylation of death-asscciated protein kinase (DAPK) gene in blood from non-small cell lung cancer (NSCLC) patients and to evaluate its clinical significance. Methods: A methylation-specific PCR was performed for the detection of promoter hypermethylation of DAPK gene in blood DNA from 65 cases of NSCLC patients,and the relation of the aberrant methylation of DAPK gene and the clinical pathological data was analyzed. Results: 30.8% (20/65) of the blood from 65 cases of NSCLC patients showed hypermethylation in DAPK promoter, whereas no methylated DAPK gene promoter was found in blood from normal control or patients with benign lung diseases. DAPK gene promoter methylation in blood did not strongly correlate with the pathological class, stage, metastasis and differentiation of NSCLC. Conclusion: Detection of the aberrant methylation of DAPK gene in blood DNA from NSCLC patients might offer an effective means for the earlier auxiliary diagnosis of the malignancy.

Objective To investigate the effect of hyperbaric oxygen to reduce stress-induced gastric mucosal injury in rabbits with blast-related brain injury.Methods Posttraumatic stress ulcer in a rabbit model of blast-related brain injury was created using paper detonators.Ninety New Zealand white rabbits were randomly divided into normal control group (n =10),blast-related injury group (n =40) and hyperbaric oxygen treatment group (hyperbaric oxygen group,n =40) according to the random number table.Rabbits in blast-related injury group and hyperbaric oxygen group were subgrouped at 6 hours,24 hours,72 hours and 7 days postinjury,with 10 rabbits at each time point.Gastric mucosal injury was detected in aspects of ulcer index,TUNEL method for apoptotic index,and alcian blue-periodic acid-schiff (AB/PAS) staining and semi-quantitative analysis of pathological histology using.Results Neither stress ulcer nor apoptosis occurred in normal control group.At 6 hours,24 hours,72 hours and 7 days postinjury,index of gastric ulcer in blast-related injury group was 12.08 ± 1.02,25.56 ± 0.88,20.36 ±0.96 and 17.56 ± 1.12 respectively,far higher than 8.02 ±0.58,15.22 ± 1.05,12.39 ±0.65 and 9.22 ± 0.79 in hyperbaric oxygen group (P ＜ 0.05) ; apoptotic index in blast-related injury group was 20.22 ± 1.56,27.56 ± 0.96,24.36 ± 1.23 and 16.38 ± 1.24,far higher than 10.78 ± 0.93,13.89 ±0.84,10.55 ± 1.58 and 8.56 ± 1.47 in hyperbaric oxygen group (P ＜ 0.05).Thickness of AB/PAS-positive mucin layers showed marked attenuation even loss in blast-related injury group,but the loss was minor in hyperbaric oxygen group (P ＜ 0.05).Conclusion Hyperbaric oxygen is effective to decrease the stress ulcer index and apoptosis index of gastric mucosa,increase the thickness of AB/PAS positive layer and lighten the gastric mucosal injury of rabbits with blast-related brain injury.

Objective To investigate the effects of acidosis on the current change of transient receptor potential channel M7 (TRPM7) and collagen production in mouse cardiac fibroblast (MCFs), and to explore the pathophysiological function of TRPM7 on the cardiac fibrosis. Methods (1) The model of MCFs was established and isolated. (2) MCFs was subcuhured. (3) Patch clamp technique was used to observe the current characteristics of TRPM7 in low PH solutions. (4) The influence of acidic condition on Ca2+ influx in MCFs was recorded by calcium fluorescent indicators. Results (1) There was a high level expression of TRPM7 in MCFs and the electrophysiological characteristics of TRPM7-like (TRPM7L) was similar to that of TRPM7. (2) Ca2+ influx was increased in acidic condition, and the F340/F380 ratio was increased from 1.0 to 4. 6 at pH 4.0 compared with pH 7.0 (t=2.72, P<0.01). Conclusions (1) TRPM7 is the molecular basis of the native TRPM7L in MCFs and TRPM7L plays an important role in Ca2+ influx. (2) The pathophysiological function of MCFs is influenced by regulation of Ca2+ influx mediated by TRPM7L in the condition of acidosis.

Objective To investigate the changes of TRPM 7-like current in mouse cardiac fibroblast(CFs) after myocardial infarction and the effect of myocardial ischemia on the TRPM 7 expression and current. Methods (1) The model of myocardial infarction was made and CFs were isolated;(2) CFs were cultured and infected by TRPM 7 siRNA;(3) The effects of myocardial ischemia on TRPM 7 current were recorded by whole cell patch clamp technique;(4) The influences of myocardial isehemia on Ca2+ influx in CFs were recorded by Ca2+ fluorescence imaging. (5) The effects of ischemia on total collagen content of CFs were studied. Results (1) Ca2+ inward current of CFs was increased after myocardial infarction [(16.2±1.7) vs. (7.4±0.7) pA/pF, P<0.053];(2) TRPM 7 current was reduced by 90% after siRNA infection;(3) The total collagen content of CFs after ischemia was approximately 2.3-fold higher than basic value. Conclusions Ca2+ influx in CFs plays an important role in the pathophysiological process of myocardial fibrosis.

Objective To investigate the value of combined detection of serum CA125 ,VEGF‐C andβ2‐MG levels on early diag‐nosis of retroperitoneal lymph node metastasis in the patients with ovarian cancer .Methods Fifty‐one patients with ovarian cancer undergoing retroperitoneal lymph node cleaning operation were included as the experimental group ,including 29 cases of positive retroperitoneal lymph node metastasis .Contemporaneous 32 cases of benign ovarian tumor were selected as the control group .The serum CA125 level was detected by using electricity chemiluminescence (electrocheminescence) ,serum VEGF‐C level by using en‐zyme‐linked immunosorbent assay (ELISA) and serumβ2‐MG level by using the latex enhanced immune turbidimetric method .The serum CA125 ,VEGF‐C andβ2‐MG levels were compared among various groups .Results The serum CA125 ,VEGF‐C andβ2‐MG levels in the experimental group were (1 682 .5 ± 261 .5)μg/mL ,(2 125 .6 ± 96 .7)pg/mL and (2 .52 ± 0 .61)mg/L respectively , which in the control group were (30 .5 ± 6 .3)μg/mL ,(1 738 .0 ± 79 .8)pg/mL and (1 .87 ± 0 .56)mg/L respectively ,the difference between them was statistically significant (P< 0 .01) .The serum CA125、VEGF‐C、β2‐MG levels in 29 cases of retroperitoneal lymph node metastasis were more significantly higher ,which were (1 997 .3 ± 376 .8)μg/mL ,(2 895 .2 ± 126 .8)pg/mL and (4 .95 ± 0 .69)mg/L respectively ,and the difference was statistically significant compared with the control group (P<0 .01) .The sensitivity ,specificity and accuracy rate of the combined detection of serum VEGF‐C ,β2‐MG and CA125 for diagnosing ovarian cancer retroperitoneal lymph node metastasis were 95 .8% ,97 .3% and 98 .5% respectively .Conclusion The combined detection of serum VEFG‐C ,β2‐MG and CA125 has an important clinical value in early diagnosing retroperitoneal lymph node metastasis of o‐varian cancer .

Adipose tissue is a readily available source of adult stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Peptide hydrogel is a novel biomaterial which provides three-dimensional microenvironments for a variety of cells for tissue grafting. In this study, adipose-derived stem cells (ADSCs) were isolated from rats, seeded into the peptide hydrogel polymer scaffolds and cultured in Neurobasal (NB) media supplemented with B27, basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). Ten days after the culture, some cells were expanded into clonal populations in which the expression of both Nestin and Brdu was detected but only Brdu expression was detected in the cells that were not expanded into clonal populations. Our results suggested that ADSCs in peptide hydrogel polymer scaffolds can be induced to differentiate into cells capable of expressing the neuron-associated markers, self-renewal and self-propagation.

Objective To study the nutritional risks in elderly patients with prostate cancer and explore its related factors.Methods 346 elderly patients with prostate cancer in Wuhan area from May 2013 to May 2014 were chosen as the objects in this study.Nutritional risk screening 2002 was used to evaluate nutritional risk.The sleep quality,anxiety,lower urinary tract symptoms,differentiation and other data of patients were collected.The influencing factors for nutritional risk in the patients were analyzed.Results The ratio of nutritional risk in elderly advanced prostate cancer patients was 35.3％ (122/346).The nutritional risk was increased with age (P＜0.05).The prostate cancer patients with nutritional risk had a higher international prostate symptom score (IPSS) (P＜ 0.05).Monovariate factor analysis showed that sleep quality,operation,pathology staging,radiotherapy,chemotherapy,lower urinary tract symptoms (LUTS) were the impact factors for nutritional status in elderly patients with prostate cancer (P＜0.05).Logistic regression analysis showed that age (OR=0.29),sleep quality (OR=0.25) were the protective factors for the nutritional status,while surgery (OR=12.67),pathological staging (OR=1.65),radiotherapy (OR=2.65),SPSS (OR=1.55),chemotherapy (OR=1.85) were the risk factors for nutritional status (P＜ 0.05).Conclusions The incidence of nutritional risk is high in elderly prostate cancer patients.Age,sleep quality are the protective factors,and operation,pathology staging,SPSS,chemotherapy,and radiotherapy are the risk factors for nutritional status.