OBJECTIVETo observe the effect of Wenyang Huoxue Lishui Recipe (WHLR) containing serum on the expression of cathepsin L (CatL) in puromycin aminonucleoside-induced injury of mouse glomerular podocytes.

METHODSMouse podocyte cells (MPCs) in vitro cultured were divided into the normal control group, the model group, the dexamethasone (DEX) group, 10% WHLR containing serum group, 20% WHLR containing serum group, the vehicle serum control group. MPCs in the normal control group were cultured at 37 degrees C culture solution for 24 h. 45 mg/L puromycin was acted on MPCs in the model group for 24 h. On the basis of puromycin intervention, 1 limol/L DEX was co-incubated in MPCs of the DEX group for 24 h; 10% or 20% WHLR containing serum was co-incubated in MPCs of the 10% WHLR containing serum group and 20% WHLR containing serum group for 24 h. The vehicle serum control group was also set up by incubating with WHLR containing serum alone for 24 h. The expression of CatL and its substrate Synaptopodin in podocytes were detected by cell immunofluorescence staining. FITC-conjugated phalloidin was used to stain F-actin. A cortical F-actin score index (CFS index) was designed to quantify the degree of cytoskeletal reorganization in cultured podocytes.

RESULTSCompared with the normal control group, the expression of synaptopodin significantly decreased and the expression of CatL significantly-increased in the model group. F-actin arranged in disorder, gradually forming pericellular F-actin ring. CFS index was obviously elevated (P < 0.01). Compared with the model group, the epression of synaptopodin increased, the expression of CatL decreased, and CFS index also decreased in the DEX group, 10% WHLR containing serum group, and 20% WHLR containing serum group (P < 0.05, P < 0.01). Compared with the DEX group, the expression of synaptopodin decreased in 10% WHLR containing serum group, CFS index also decreased in 20% WHLR containing serum group (P < 0.05).

CONCLUSIONSWHLR could up-regulate the expression of synaptopodin, down-regulate the expression of CatL, and alleviate cytoskeletal reorganization of F-actin. It was helpful to stabilize the cytoskeleton of F-actin and improve the merging of podocytes.

Actins ; metabolism ; Animals ; Cathepsin L ; metabolism ; Cells, Cultured ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Kidney Glomerulus ; cytology ; Mice ; Microfilament Proteins ; metabolism ; Podocytes ; drug effects ; pathology ; Puromycin Aminonucleoside ; adverse effects ; Up-Regulation

The involvement of astrocytes and correlation between neuronal injury and astrocyte response were studied. Blockingmiddle cerebral artery and reperfusing o. 5～48 h, H-E staining, immunoccytochemistry single-and double-labeling, dotble label-ing combined with TUNEL and GFAP immunocytochemistry were used to investigate neuronal injury and astrocyte response.The is chemic area peaked at 24 h of reperfusion. The neurons presented irreversible degeneration at 6 h of reperfusion. At24 h,ischemic area in the preoptic area developed into infarcted area; astrocytes exhibited differential morphological features: reactive,malnourished and degenerative changes. At 48 h of reperfusion, the number of astrocytes began to go up. The astrocytes in is-chemic area didn't proliferate within 48 h. By contrast, a few astrocytes underwent apoptosis. In conclusion, these data indicatethat the reaction of astrocytes is closely connected with the extent of neuronal injuries. The reactive astrocytes imply that astro-cytes positively respond to the neuronal injuries, which might play a role in promoting neuronal survival.

Aim To investigate the effects of meloxi-cam on the proliferation,migration and expression of PTEN of human colorectal cancer LoVo cells.Meth-ods The colony formation test was used to detect the effect of meloxicam on the proliferation of LoVo cells. The cell migration assay was applied to analyze the effect of meloxicam on LoVo cells activity.The RT-PCR assay was used to detect the effect of meloxicam on the mRNA expression of PCNA and PTEN gene. The western blot assay was applied to analyze the effects of meloxicam on the expression of PTEN pro-tein.The recombinant adenovirus and Annexin-V assay were used to testify the relationship between PTEN gene and anti-cancer effect of meloxicam.Results Compared with the control group,meloxicam could in-hibit the colony formation and PCNA protein expression of LoVo cells.At 48 h and 80 μmol·L -1 ,the expres-sion of PCNA protein was reduced to 61 .57% ± 2.81 %(T =7.086,P =0.01 9),the mRNA expres-sion of PTEN gene increased to 1 60.43% ±4.71 %(T=24.244,P =0.002),and the expression of PTEN protein increased to 1 52.63% ±3.33%(T =27.359, P =0.001 ).Results Annexin-V test indicated that the anti-cancer effect of meloxicam was associated with the up-regulated expression of PTEN.Conclusions Meloxicam can inhibit the proliferation and migration of LoVo cells by up-regulating the expression of PTEN.

Objective To explore the different probability of carcinoma and the necessity of treatment in cervical columnar ectopy (CCE),normal smooth cervix and cervical intraepithelial neoplasia(CIN) by investigating the different expression of matrix metalloproteinase-9(MMP-9).Methods The expression of MMP-9 was detected by using immunohistochemical method in 30 cases of CCE,28 cases of CIN and 30 cases of normal smooth cervix from 2007 to 2012.Results The positive expression of MMP-9protein in CIN was 96.4％ (27/28),CCE was 23.3％ (7/30) and normal smooth cervix was 20.0％ (6/30)respectively.The positive expression of MMP-9 protein was higher in CIN than CCE and normal smooth cervix,the difference has statistical significance (P＜ 0.01),there was no significant difference between CCE and normal smooth cervix (P ＞ 0.05).Conclusions CCE,which was a physiological performance of cervical erosion,should not be overtreated.But the women of childbearing age should be routinely performed in cervical cancer screening.

Aged ; Cardiac Pacing, Artificial ; Cardiac Resynchronization Therapy ; Female ; Heart Ventricles ; Humans

BACKGROUND: This study aimed to observe the effect of early goal directed therapy (EGDT) on tissue perfusion, microcirculation and tissue oxygenation in patients with septic shock. METHODS: Patients with early septic shock (<24 hours) who had been admitted to the ICU of Zhongda Hospital Affiliated to Southeast University from September 2009 through May 2011 were enrolled (research time: 12 months), and they didn't meet the criteria of EGDT. Patients who had one of the following were excluded: stroke, brain injury, other types of shock, severe heart failure, acute myocardial infarction, age below 18 years, pregnancy, end-stage disease, cardiac arrest, extensive burns, oral bleeding, difficulty in opening the mouth, and the onset of septic shock beyond 24 hours. Patients treated with the standard protocol of EGDT were included. Transcutaneous pressure of oxygen and carbon dioxide (PtcO2, PtcCO2) were monitored and hemodynamic measurements were obtained. Side-stream dark field (SDF) imaging device was applied to obtain sublingual microcirculation. Hemodynamics, tissue oxygen, and sublingual microcirculation were compared before and after EGDT. If the variable meets the normal distribution, Student's t test was applied. Otherwise, Wilcoxon's rank-sum test was used. Correlation between variables was analyzed with Pearson's product-moment correlation coefficient method. RESULTS: Twenty patients were involved, but one patient wasn't analyzed because he didn't meet the EGDT criteria. PtcO2 and PtcCO2 were monitored in 19 patients, of whom sublingual microcirculation was obtained. After EGDT, PtcO2 increased from 62.7±24.0 mmHg to 78.0±30.9 mmHg (P<0.05) and tissue oxygenation index (PtcO2/FiO2) was 110.7±60.4 mmHg before EGDT and 141.6±78.2 mmHg after EGDT (P<0.05). The difference between PtcCO2 and PCO2 decreased significantly after EGDT (P<0.05). The density of perfused small vessels (PPV) and microcirculatory flow index of small vessels (MFI) tended to increase, but there were no significant differences between them (P>0.05). PtcO2, PtcO2/FiO2, and PtcCO2 were not linearly related to central venous saturation, lactate, oxygen delivery, and oxygen consumption (P>0.05). CONCLUSION: Peripheral perfusion was improved after EGDT in patients with septic shock, and it was not exactly reflected by the index of systemic perfusion.

Objective To construct two DNA vaccines encoding Gag-Env fusion protein and Tat-Rev-Integrase(C-half)-Vif-Nef fusion protein derived from the first HIV-1 CRF01_AE isolate(AE2f) in Chi-na and to evaluate the immunogenicity in mice. Methods Two DNA vaccines were constructed by inserting the codon optimized and synthesized gag-env fusion gene and tat-rev-integrase(c-half)-vif-nef fusion gene de-rived from AE2f into mammalian expression vector pDRVISV1. 0, the generated DNA vaccines were desig-nated as pSVAE/GE and pSVAE/TRIVN, respectively, and their in vitro expression were determined by Western blot with transfected 293T cells. Mice were i. m. immunized with either pDRVI1.0 as mock control, pSVAE/GE or pSYAE/TRIVN for 4 times at two-week interval. Two weeks following the final im-munization, cellular responses to pool of HIV-1 Env, Gag, Tat, Rev, Intergrase, Vif and Nef peptides were evaluated by ELISPOT assay. Results The construction of DNA vaccine pSVAE/GE and pSVAE/TRIVN was validated by restriction enzyme digestion and bidirectional sequencing. Western blot showed a specific band at molecular mass 220×10~3 in lane of pSVAE/GE transfeeted 293T cell and a specific band at 95×10~3 in the lane of pSVAE/TRIVN. Both DNA vaccines mounted significant specific T cell responses with (3010 ± 566) SFC/10~6 splenocytes for DNA vaccine pSV AE/GE and (948 ± 737) SFC/10~6 spleno-cytes for DNA vaccine pSVAE/TRIVN, whereas the mock control of pDRVISV1.0 only raised marginal T cell responses. Conclusion Both pSVAE/GE and pSVAE/TRIVN were capable of expressing the inserted fusion immunogen genes and able to elicit vigorous cellular immune responses, therefore, these DNA vac-cines are highly immunogenic.

Objective To evaluate the effect of comprehensive control for schistosomiasis with emphasis on environmental modification in the Bianmin River water system of Nanjing City.so as to provide scientific evidence for making up further control measures in this water system.Methods Schistosome infections of Oncomelania snails in the waterway.sentinel mice in water and neighbouring human were investigated longitudinally from 1998 to 2007,and the changes of huaman infection rates in differentyears,the infection rates of sentinel mice and snails in different settings were analyzed and compared.Results A total of 77 395 snails collected from the Bianmin River water system were dissected from 1998 to 2007,and among them,27 snails were infected with Schistosoma japonicum,with a total snail infection rate of 0.03%.A total of 61 039 snails collected from the neighbouring marshland which connected to the Yangtze River wore dissected,and among them,257 were infected with S.japonicum,with a total snail infection rate of 0.42%,and there was a significant difference compared with that in the water system(χ~2=248.55,P<0.01).After the protection works in the waterway,the infection rates of sentinel mice in the water system decreased from 69.68% in 1998 to 17.50% in 2001.with a reduction rate of 74.89%.Two years afterthe clearance ofmarshlandinthewaterway,no infected sentinel mouse was found.The infection rates of residents from 1998 to 2007 were 1.96%,1.37%,1.34%,1.60%,0.30%, 0.26%,0.16%,0.10%,0.04% and 0,respectively,andthe rates declined year by year afterthecomprehensive control.Conclusions The control measures based on the elimination of snail habitats in the waterway that is connected to the Yangtze River have achieved obvious effect.However,the clearance of the re-emerging snail habitats should be carried out termly to consolidate the control effect.

Objeetive To analyze traffic accidents involving pedestrians in Chongqing from 2000 to 2006 so as to understand injury characteristics of the pedestrians. MethodsThe data of pedestrian road traffic crashes and injuries in three districts including one district in downtown,one in suburb and one in county of Chongqing from 2000 to 2006 were collected from road traffic crash and road traffic injury database to analyze severity of crashes,casualties,action and location of pedestrians in crashes. Results The road crashes led to 7 934 pedestrian injuries (24.74％) and 867 pedestrian deaths (49.46％).Intensity of the pedestrian road crashes in rural areas was higher than that in downtowns and suburbs.Of all pedestrian casualties,57.91％ were due to illegally crossing the driveway,and 15.43％ of the casualties were on the crosswalk.Most of the casualties occurred on the main roads,the third class highways and second main roads in Chongqing.But serious crashes occurred on the first and second class highways and substandard highways,which led to average 3.17,4.13 and 5.42 deaths,respectively.Pedestrians accounting for 52.46％ of the total severe injuries were more than the proportion among the minor to moderate injuries.Head injuries were the most common for the pedestrians and chest injuries were also usual among the severe injuries. Conclusions The intensity of pedestrian road crashes is higher than that of other crashes,especially in rural areas.It should be noticed that the main injuries of the pedestrians are head injuries,which is mainly resulted from illegally crossing driveway.It is worth noticing that a rather high proportion of pedestrians are impaired on the crosswalks,indicating that we should pay more attention to protect right of the pedestrians on the crosswalks during transportation law enforcement and safety education in the future.

Objective To observe the effects of recruitment maneuver (RM) and tidal volume with different amount of gas after RM ventilation on lung diastole function in rats with acute lung injury (ALI). Method ALI rat models were induced by intravenous infusion of lipopolysaccharide (LPS) in dose of 6 mg/kg. Twenty-five rats were randomly(random number) divided into control group ( n = 5), ALI group ( n = 5), low tidal volume group (LV group,VT= 6 mL/kg, n = 5), sustained inflation (SI) with low tidal volume (SI+ LV group, VT=6 mL/kg, n = 5), and SI with moderate tidal volume group (SI+ MV group, VT= 12 mL/kg, n = 5). The RM carried out by using SI with airway pressure 30 cmH-2O for 30 seconds, and the positive end-expiratory pressure (PEEP)was set at 5 cmH2O. Lung tissue was taken after mechanical ventilation for 5 hours. The mean arterial pressure (MAP) was monitored throughout the entire course of experiment. Endothelin-1 ( ET-1 ), endothelial nitricoxide synthase (eNOS), and acetylcholine-(Ach-) induced endothelium-dependent relaxation response of isolated pulmonary artery rings were investigated after mechanical ventilation for 5 hours. Results The LPS increased the ET-1 level in lung tissue, decreased the level of eNOS in lung tissue, and impaired the Ach-induced endothelium-dependent relaxation response in pulmonary vassals, without obvious influence on systemic hemodynamics. SI + LV significantly reduced LPS-induced elevation of ET-1 level, and increased the level of eNOS, and significantly lessened endothelial dysfunction and ameliorated dysfunction od endothelium-dependent relaxation in pulmonary vas sals. Conclusions RM with high tidal volume or lowtidal volume ventilation could improve the lung vascular endothelial function of rats with acute lung injury, and RM with low tidal volume ventilation could lessen more the injury of lung vascular endothelial diastole function in rats with acute lung injury.