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

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.

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.

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 value of echo‐contrast RT‐3DE for assessment of left ventricular volume and function in patients with left ventricular non‐compaction(LVNC) .Methods Twenty‐one patients of LVNC were involved and underwent non‐enhanced and contrast‐enhanced RT‐3DE to evaluate left ventricular end‐diastolic volume (LVEDV) ,left ventricular end‐systolic volume (LVESV) ,left ventricular ejection fraction (LVEF) .The endocardial border definition of LV was graded for each of the 16 LV segments as follows :0 = border invisible ,1 = border visualized only partially ,and 2 = complete visualization of the border .Three image‐quality groups (good ,fair ,and uninterpretable) were identified . Results ①Duringcontrast‐enhancedRT‐3DE,ascomparedwithnon‐enhancedRT‐3DE,thenumberof segments with complete visualization of the endocardial border increased significantly (55% vs 82% ,P <0.01) ,and the number of patients with a good‐quality echocardiogram increased significantly (33% vs 81% , P <0.01) .②Contrast‐enhanced RT‐3DE provided significantly larger values of LVEDV ( P < 0 0.1) and LVESV ( P < 0 0.1) as compared with non‐enhanced RT‐3DE ,the values of LVEF were not statistically different between the two techniques ( P =0.07) .③Intra‐and inter‐observer agreement for assessment of LV volumes and systolic function improved during contrast‐enhanced RT‐3DE ,as compared with non‐enhanced RT‐3DE .Conclusions Contrast‐enhanced RT‐3DE can increase the prevalence of good‐quality echocardiograms and significantly improve the reproducibility of LV volumes and function measurements .

The present work is to investigate the chemical constitutions of Hyptis rhomboidea and their antifungal activities. The compounds were isolated by Toyopearl HW-40, Sephadex LH-20, MCI-Gel CHP-20, RP-18, PTLC and silica column chromatographic methods and subjected to evaluate some monomers antifungal activity of eight kinds of plant pathogenic bacteria. Eleven compounds were isolated and identified as ethyl caffeate (1), ursolic acid (2), oleanolic acid (3), vanillactic acid (4), methyl rosmarinate (5), kaempferol 3-O-alpha-L-rhamnopyranosyl-(1 --> 6) -beta-D-glucopyranoside (6), kaempferol 3-O-alpha-L-rhamnopyranosyl-(1 --> 6)-beta-D-glucopyranoside (7), ilexgenin A (8), beta-amyrin (9), kaempferol 3-O-beta-D-glucopyranoside (astrgalin, 10) and cholest-5-ene-3beta, 4beta-diol (11). Compound 1 showed the strongest inhibitory effect on Sclerotinia sclerotiorum with the MIC 16.2 mg x L(-1), and compound 5 showed the strongest inhibitory effect on S. minor and Exserohilum turcicum with MIC 16.2, 8.1 mg x L(-1), respectively. All compounds were isolated from the H. rhomboidea for the first time, and compounds 1 and 5 showed antifungal activity.
Antifungal Agents ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Fungi ; drug effects ; Hyptis ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo observe the clinical effect of acupuncture on acute heart failure (AHF) patients.

METHODSTotally 60 patients who were diagnosed as AHF were assigned to the acupuncture group and the control group, 30 in each group. Those in the control group received inotropic agents, preload and afterload reducing therapy, anti-infection and so on. Besides, those in the acupuncture group received needling at relative points, once daily for 5 consecutive days. The mean arterial pressure (MAP), heart rate (HR), cardiac index (CI), stroke volume index (SI), left ventricle working index (LCWI) were monitored by thermodilution pulse-indicated continuous cardiac output (PiCCO) technique. Changes of the aforesaid data were compared between before and after treatment. The ICU length of stay, readmission rate,and the 28-day mortality were also compared between the two groups.

RESULTSAfter 5 days of the treatment, CI, SI, and LCWI increased more obviously (P < 0.01), HR and MAP decreased significantly (P < 0.05). Of them, CI, SI, and LCWI increased more obviously in the acupuncture group (P < 0.05).There was no obvious difference in HR or MAP between the two groups (P > 0.05). Compared with the first day of admission in the same group, CI, SI, and LCWI obviously increased in the acupuncture group from the second day (P < 0.05). HR started to decrease since the fourth day (P < 0.05), and MAP began to decrease until the fifth day (P < 0.05). CI, SI, and LCWI started to increase in the control group from the third day (P < 0.05); HR and MAP both began to decrease since the fifth day (P < 0.05). Compared with the control group, the ICU length of stay was obviously shortened in the acupuncture group (P < 0.05). The readmission rate and the 28-day mortality rate were lower than those of the control group (P < 0.05).

CONCLUSIONThe combination of acupuncture and Western medical therapy might strengthen acute heart failure patients' heart functions, elevate the therapeutic effect, and improve the prognosis.

Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Female ; Heart Failure ; physiopathology ; therapy ; Heart Rate ; Humans ; Male ; Middle Aged ; Stroke Volume