Objective To investigate the role of PTEN protein in secretion of collagen Ⅳ and fibronectin after stimulation of transforming growth factor beta 1(TGF-?1) from renal fibroblasts of rat in vitro.Methods The cultured rat renal fibroblasts were transfected with the reconstructed adenovirus containing PTEN or adenovirus only containing green fluorescence protein(GFP).The fibroblsts were treated in four manners:control group with no added treatment,TGF-?1 group with TGF-?1 stimulation,PTEN+ TGF-?1 group with TGF-?1 stimulation after Ad-PTEN transfection,and GFP+ TGF-?1 group with TGF-?1 stimulation after Ad-GFP transfection.Invert fluorescent microscope was used to detect the GFP expression,meanwhile the PTEN mRNA was determined by RT-PCR method.36h after the transfection,TGF-?1 was added into the culture medium in a concentration of 10ng/ml.After another 24h,ELISA method was used to evaluate the level of collagen Ⅳ and fibronectin.Results The expressions of both GFP and PTEN mRNA increased obviously after the rats' renal fibroblasts were transfected with adenovirus.The secretion of collagen Ⅳ and fibronectin increased significantly in both TGF-?1 group and GFP+ TGF-?1 group compared with that in control group,and decreased markedly in PTEN+ TGF-?1 group compared with that in both TGF-?1 group and GFP+ TGF-?1 group(P

Objective To investigate the stimulating effects of activin A(ACT-A) and follistatin(FS) on the secretion of type I collagen(Col I) and the expression of Col I mRNA of cultured rat renal interstitial fibroblasts in vitro.Methods Rat renal interstitial fibroblasts isolated from normal SD rat renal medulla were cultured in vitro.The cells were divided into three groups:different concentrations of ACT-A(group A),different concentrations of FS(group F),and different concentrations of FS plus 30ng/ml ACT-A(group A+F).The expression of Col I mRNA in cultured rat renal interstitial fibroblasts was detected by RT-PCR,and the expression of Col I protein in cultured rat renal interstitial fibroblasts was examined by immunocytochemistry.Results Renal interstitial fibroblasts were successfully cultured in vitro.In present study,the expression of Col I mRNA increased significantly in cultured rat renal interstitial fibroblasts after stimulated by ACT-A in dose-dependent manner(P0.05).FS could inhibit the effects of ACT-A(30ng/ml) on Col I mRNA and Col I protein of the cultured renal interstitial fibroblasts in dose-dependent manner in A+F group(P

Objective To study the mutagenicity of water from Yangling-Xingping section of Weihe River and wastewater from paper mills along the river. Methods No1 and No2 paper mill as the pollution source were located along the Yangling-Xingping section of Weihe River,their sampling points were named paper mill 1 and paper mill 2.The outlets for sewage at the bank of Weihe River in Yangling and Xingping were selected the sampling points of pollution sections, namely Yangwu and Xingwu respectively. The clear sections (Yangshang and Xingshang) were situated at a distance of 1 km from polluted sections at the upper reaches of Weihe River were selected and at the sames at the lower reaches were selected as the self-purification sections, namely Yangxia and Xingxia. The mutagenicity of water samples collected from above sampling points was tested by Ames test,mouse sperm malformation test, Vicia faba root tips cell micronucleus test. Results The results of Ames test showed that the water samples of Xingwu, Xingxia and two paper mills presented mutigenicity.The water samples of Xingwu, Xingxia and two paper mills showed significantly higher levels of sperm malformation rates compared with that of negative control group(P

Objective To establish the electric controlled cortical impact (eCCI)-induced traumatic brain injury (TBI) model in rats with different severity in degree,which may serve as a suitable platform to provide experimental evidence for the pathophysiological following TBI.Methods A total of 40 male Wistar rats were randomly divided into 3 experimental groups and sham group.TBI rats (n=10/group) were positioned beneath the controlled cortical impactor device (eCCI) and subjected to impact injury at 2 mm depth of penetration,for a sustained depression of 200 ms,at 4 m/s,5 m/s,6 m/s velocity for mild,moderate,and severe TBI,respectively.Sham-operated rats (n=10) underwent identical surgical procedures,including craniotomy,without receiving the cortical impact.Neurological function and regional cerebral flow (24 h after CCI),contusion volume,histopathological,and ultrastructural changes (48 h after CCI) were measured,respectively.Results The severity of the pathological changes in rats was increased as the injury aggravated.The eCCI device impacted the brain at 4 m/s,5 m/s,6 m/s velocity for mild,moderate,and severe TBI,respectively.TBI groups showed impaired neurological function,and decreased rCBF lower than that of sham-operated group (all P＜0.01).Furthermore,neuronal pathological abnormalities in TBI groups,including neuron shrinking,perineuronal vacuole,and structural abnormalities of mitochondria.Increased severity of injury was apparent following the increased level of the impacted velocity,and significant differences were observed between TBI groups (P＜0.05).Conclusion The TBI animal model with mild,moderate,and severe brain injury can be established successfully by 4 m/s,5 m/s,and 6 m/s of impact velocity respectively with the eCCI-6.3 device.The novel eCCI-induced TBI model in rats possibly serves as a novel useful approach in the development of TBI models.

Objective To study the effect of brain-derived neurotrophic factor (BDNF) on the environmental nutrition and neural differentiation of the transplanted stem cells under hypothermia.Methods The BDNF gene mediated by liposome was transfected into 293T cell line, and ELISA assay was applied to find the peak time of BDNF expression. When BDNF was highly expressed, the supernatant was collected for establishment of SD rat models of brain injury. The rats were divided into Group A (stem cell transplantation group) and Group B (stem cell transplantation and BDNF group). Rats in both groups were under hypothermia treatment for five days. Four and eight days later ( three days from rewarming), rat brain tissues were obtained to detect the expressions of proliferating cell nuclear antigen (PCNA), nestin, neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) by immunohistochemical method and to detect the apoptosis by in situ hybridization. Finally, the nerve function scores were obtained for evaluation of the nerve function. Results The ELISA showed that the high level of BDNF expression was at 48 to 60 hours after gene transfection. PCNA and nestin were highly expressed, while NES and GFAP showed nil or low level of expression in both groups at the fourth day after hypothermia, with little apoptotic cells especially in the Group B (P ＜0.05). The expressions of PCNA and nestin were decreased, but the expressions of NSE and GFAP were increased at the third day after rewarming. The positive rate of NSE expression in the Group B was much higher and the apoptotic cells were much less compared with the Group A ( P ＜ 0. 05 ). A better nerve score was obtained in the Group B. Conclusion BDNF can enhance the survival rate of the transplanted stem cells and induce their differentiation into neurons under hypothermia.

ObjectiveTo investigate the roles of 11 β-hydroxysteroid dehydrogenase type 1 ( 11 β-HSD1 )on hippocampus of rat with chronic unpredictable mild stress (CUMS).MethodsTwenty-four male SpragueDawley rats were randomly divided into control group and depressive model group. Chronic unpredictable mild stress (CUMS) was used to make up depressive animal model.Behavioral changes were recorded by body weight measuring,sucrose consumption test (SCT) and open field test (OFT),respectively.The mRNA transcription of 11β-HSD1 in hippocampus tissues of the rats were detected by real-time RT-PCR,and the protein expression of 11β-HSD1 were detected by western blot and immunofluorescence.ResultsBcforc starting CUMS protocol,the rats exhibited equivalent weight and sucrose consumption.Twenty-eight days after CUMS protocol,behavior parameters such as body weight,sucrose consumption,nunber of crossing,and number of rearing were significantly decreased in rats exposed to CUMS group compared with control group (P ＜ 0.05,P ＜ 0.01 ).Correspondingly,realtime RT-PCR assays showed the mRNA expression of 11 β-HSD1 in the hippocampus of CUMS group,which was (31 ±9) ％ lower than that of control group.Meanwhile,the protein expression of it in CUMS group was lower than that of control group (P ＜ 0.05 ).Inmunofluorescence revealed that the number of positive 11 3-HSD1 cells was high (223 ± 13) in the control group,while the number was decreased prominently (92 ± 11 ) in the CUMS group (P ＜ 0.01 ).ConclusionDepressive behavior of rats is induced and the expression of 11 β-HSD1 in the hippocampus is decreased prominently by CUMS,the mechanism of which is at least related to the low expression of 11β-HSD1 and disturbance of glucocorticoid metabolism caused by CUMS.

To study the correlation between traditional Chinese medicine (TCM) syndrome type and the ultrasound imaging changes in patients with limb lymphedema, and to provide evidence for TCM syndrome differentiation.

Objectives To understand essential basis data availability for implementation of cost accounting in TCM hospitals. Methods Costs of a total of 1588 TCM hospitals were investigated by conducting the baseline survey via questionnaires in 2013. Results Departmental basic data (including ratio of office staff and entire staff, ratio of outpatient and inpatient departments in clinical department), and construction data for housing were well accessible, while utility fee and internal service data were not. Conclusion ①Public TCM hospitals need to improve and specify the basic cost data. ②Differences exit among different types and levels of hospitals in the availability of data. ③There are still difficulties in carrying out cost accounting in public TCM hospitals, which needs strong supports from all relevant departments.

A near infrared spectroscopy ( NIRS) method was used for rapid quality evaluation of ferulic acid content in chrysanthemum morifolium cv. ( Chuju) continuous cropping soil. Standard leverage, studentized residual and Mahalanobis distance were calculated to eliminate abnormal samples. After the initial near infrared spectrum was treated by two second derivative and Norris smoothing filter noise, 6000-4000 cm-1 wave number range and 7 factors were chosen for partial least squares ( PLS) calibration model. The results showed that good correlation was presented between the calibration set/validation set and the values determined by high performance liquid chromatography, and the calibration correlation coefficient ( Rc ) and validation correlation coefficient ( Rcv ) were 0. 9914 and 0. 9935, respectively. Root mean square error of calibration (RMSEC), root mean square error of validation (RMSEP) and root mean square error of cross-validation (RMSECV) were 0. 484, 0. 539 and 0. 615, respectively. This method was accurate, reliable, simple, rapid and nondestructive, and could be applied to the fast analysis for ferulic acid in continuous cropping soil.

Purpose To investigate the expression of insulin-like growth factorIImRNA binding protein 3 ( IMP3 ) and microvessel density (MVD) in squamous carcinoma of the cervix (SCC) and analysis their relationship in SCC. Methods The expression of IMP3 and MVD was examined by immunohistochemistry SP method in normal cervical epithelium ( NCE) , low-grade cervical intraepi-thelial neoplasia (CIN-L), high-grade cervical intraepithelial neoplasia (CIN-H) and SCC. Results (1)The positive expression rates of IMP3 in NCE, CIN-L, CIN-H, SCC tissues were 0(0/15), 0(0/11), 37.5%(9/24) and 86.0%(43/50), the difference was statistically significant (x 2 =53.345, P=0.000). IMP3 expression was significant difference among NCE and CIN-H, SCC (P<0.008 3), and that was also among SCC and CIN-L, CIN-H (P<0.008 3). (2)The MVD count was increased with the development of cervical squamous lesion, there was significant difference among those groups (F=145.968, P<0.01), and the difference was al-so statistically significant between every two groups (P<0.05). The positive expression of IMP3 and MVD count in 50 cases of SCC tissues showed statistical difference in different pathologic grade, lymph node metastasis, and depth of tumor invasion groups ( P<0.05), but didn't in different patients’age groups (P>0.05). And the positive expression of IMP3 was closely related with MVD count in SCC tissues (rs =0.323, P<0.05). Conclusion IMP3 plays an important role in the occurrence, infiltration and metasta-sis of SCC, and the abnormal expression of IMP3 may relate with the angiogenesis of tumors.