Adolescent ; Child ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Killer Cells, Natural ; pathology ; Leukocyte Common Antigens ; analysis ; Lymphoma, T-Cell ; genetics ; immunology ; Male ; Microtubule-Associated Proteins ; genetics

Early stage diabetic nephropathy is characterized by elevated glomerular filtration. Recent studies have identified high-glucose induced p38 MAPK (p38) over-activation in mesangial cells. Mesangial hypocontractility is the major underlying mechanism, however, no ameliorating agents are currently available. We investigated the protective effects of emodin on high-glucose induced mesangial cell hypocontractility. Mesangial cells were cultured under normal (5.6 mM) and high glucose (30 mM) conditions. Emodin was administrated at doses of 50 mg/l and 100 mg/l. Angiotension II stimulated cell surface reductions were measured to evaluate cell contractility. p38 activity was detected using Western blotting. To further explore the possible mechanism of emodin, expression of the peroxisome proliferator-activated receptor gamma (PPARgamma) was measured and its specific inhibitor, gw9662, was administrated. Our results showed: (1) high-glucose resulted in a 280% increase in p38 activity associated with significant impairment of mesangial contractility; (2) emodin treatment dose-dependently inhibited high-glucose induced p38 over-activation (a 40% decrease for 50 mg/l emodin and a 73% decrease for 100 mg/l emodin), and mesangial hypocontractility was ameriolated by emodin; (3) both the PPARgamma mRNA and protein levels were elevated after emodin treatment; (4) inhibition of PPARgamma using gw9662 effectively blocked the ameliorating effects of emodin on high-glucose induced p38 over-activation and mesangial hypocontractility. Emodin effectively ameliorated p38 over-activation and hypocontractility in high-glucose induced mesangial cells, possibly via activation of PPARgamma.
Animals ; Cell Line ; Cell Physiological Phenomena/drug effects ; Emodin/*pharmacology ; Gene Expression/drug effects ; Glucose/*metabolism ; Mesangial Cells/cytology/*drug effects ; PPAR gamma/genetics/*metabolism ; Protein Kinase Inhibitors/*pharmacology ; Rats ; p38 Mitogen-Activated Protein Kinases/*metabolism

This study is to evaluate the cytotoxicity of mitomycin C (MMC) and its analogue 5-(aziridin-1-yl)-3-hydroxymethyl-1-methylindole-4,7-dione (629) as well as the effect of transfection of constitutive androstane receptor (CAR) on their biological effects. HepG2 cells were transfected with the plasmids mCAR1/pCR3 mediated by liposome. Vector pCR3 was used as control. Transfected cells were screened by G418 resistance and limiting dilution. The expressions of plasmid mCAR1/pCR3 and CYP2B6 mRNA were detected by RT-PCR; Cytotoxicities of MMC and 629 in vitro were evaluated in g2car cells and HepG2 cells by MTT method under anaerobic and aerobic conditions. mRNA expression of CAR and CYP2B6 can not be detected in HepG2 cells and HepG2/pCR3 cells but can in g2car cells. It is shown that plasmid mCAR1/pCR3 was transfected into g2car cells successfully and target CYP2B6 was transactivated by CAR. To compare with aerobic and anaerobic, the cytotoxicities of MMC and 629 to HepG2 cells and g2car cells had significantly enhanced (P < 0.05), and transfect CAR gene can improve the cytotoxicity of MMC (P < 0.05), but not 629 (P > 0.05). Furthermore, CYP2B6 is one master enzyme for the metabolism of MMC and not 629. Transfection of CAR can increase expression of CYP2B6 mRNA in HepG2 cells, and can affect cytotoxicities of MMC and 629.
Antibiotics, Antineoplastic ; pharmacology ; Aryl Hydrocarbon Hydroxylases ; biosynthesis ; genetics ; Aziridines ; pharmacology ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Death ; drug effects ; Cell Hypoxia ; Cell Line, Tumor ; Cytochrome P-450 CYP2B6 ; Humans ; Indoles ; pharmacology ; Inhibitory Concentration 50 ; Liver Neoplasms ; metabolism ; pathology ; Mitomycin ; pharmacology ; Oxidoreductases, N-Demethylating ; biosynthesis ; genetics ; Plasmids ; RNA, Messenger ; metabolism ; Receptors, Cytoplasmic and Nuclear ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transcription Factors ; biosynthesis ; genetics ; Transfection

Objcetive: To explore the treatment of long segment defect of tibia by using tensor fascia lata combined with iliac flap or deep circumflex iliac pedicle iliac flap. Methods: From February 2012 to August 2017, The People′s Hospital of Zun Yi City Bo Zhou District treated 16 patients who had long segment defect of tibia.There were 11 males and 5 females, age from 22 to 58 years old, the average age was 42 years old. Iliac flap grafting with tensor fascia lata combined with iliac flap or deep circumflex iliac pedicle was used to treat the defect of long segment of tibia. There were 4 cases with simple tibial defect and 12 cases with skin defect. The longest tibial defect was 5-8 cm. Results: In this study, four patients used iliac flaps with deep circumflex iliac pedicle, the area of flaps ranged from 2.5 cm×5.0 cm to 5.0 cm×10.0 cm, while the area of iliac flaps ranged from 5.0 cm×2.5 cm to 8.0 cm×4.0 cm. Twelve patients used grafting with tensor fascia lata combined with iliac flap, the area of flaps ranged from 5.0 cm×12.0 cm to 12.0 cm×23.0 cm, while the area of iliac flaps ranged from 7.0 cm×2.0 cm to 8.0 cm×4.0 cm. All 16 cases of bone flap were survived, fracture healing, without surgical complications. The average follow-up period was 1.5 years, the flaps had good appearance in 10 cases and was slightly bloated in 6 cases; the ankle had normal motion in 14 cases and had poor dorsal extension in 2 cases. X-ray films showed that the bone flap repaired the bone defects and reached bone healing. Conclusions Vascularized tensor fascia lata combined with iliac flap or deep circumflex iliac pedicle iliac flap grafts increase local blood supply and accelerate the process of fracture healing.

OBJECTIVETo study the effect of Shenfu Injection (SF) on the apoptosis of prostate cancer PC-3 cells and its possible mechanism.

METHODSWe divided prostate cancer PC-3 cells into a blank control group and three experimental groups, the latter treated with SF at 50, 100, and 200 microl/ml, respectively, for 24, 48, and 72 hours. Then we determined the proliferation of the cells by MTT assay, measured their apoptosis by Annexin V/PI flow cytometry, and detected the expression of P53 mRNA by RT-qPCR.

RESULTSCompared with the blank control group, the survival rates of the prostate cancer PC-3 cells in the 50, 100, and 200 microl/ml SF groups were (93.76 +/- 2.63)%, (81.21 +/- 1.80)% and (18.01 +/- 3.84)% at 24 hours, (94.67 +/-1.11)%, (78.33 +/- 2.89)% and (10.34 +/- 1.44)% at48 hours, and (91.30 +/- 0.47)%, (36.67 +/- 1.56)% and (1.33 +/- 0.32)% at 72 hours, all significantly increased in a dose- and time-dependent manner (P < 0.05). The expression of p53 mRNA was also markedly increased in all the three experimental groups at 48 hours (P < 0. 05).

CONCLUSIONSF can inhibit the proliferation and induce the apoptosis of PC-3 cells, which may due to its upregulation of the p53 mRNA expression.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; metabolism

OBJECTIVEThe aim of the study was to establish the contemporary epidemiological characteristics of Japanese encephalitis (JE) in Guizhou Province.

METHODSA retrospective study of National Notifiable Disease Reporting System (NNDRS) data from 1971 through 2009, was conducted to ascertain the geographical, seasonal, and age distributions of JE incidence in Guizhou Province, China.

RESULTSA total of 68 425 JE cases were reported in Guizhou from 1971-2009. The JE cases occurred sporadically in all 9 prefectures of Guizhou, mostly among residents of rural areas. Seasonal distribution of JE remained consistent over the period from 1971-2009 with the main transmission season starting from June to September and peaking in August. JE occurred mainly in children under the age of 15 years with peak incidence in the 0-6-year age group. Pearson's correlation analysis showed that JE vaccine distribution had a negative correlation with JE incidence rates during 1971-2009 (coefficient of correlation=-0.475, P<0.01).

CONCLUSIONOver the period of 1971-2009, the JE incidence rate had declined dramatically in terms of geographical and age distributions due to JE vaccination to children at risk.

Adolescent ; Age Distribution ; Child ; Child, Preschool ; China ; epidemiology ; Encephalitis, Japanese ; epidemiology ; mortality ; prevention & control ; Humans ; Incidence ; Infant ; Infant, Newborn ; Seasons ; Time Factors ; Viral Vaccines ; immunology

Objective To construct the silencing vector of FOS like antigen 1 (FOSL1) gene, and study the effects of FOSL1 on cell proliferation, cell invasiveness and the methylation level of PRDM10 gene in breast cancer cell line MDA-MB-231. Methods The FOSL1 silencing vector of gene pLVX-shRNA-FOSL1-shRNA was purchased. The FOSL1 silencing vector and the empty vector were separately transfected into MDA-MB-231, which were regarded as transfection group and empty group, respectively. Untransfected MDA-MB-231 was used as control group. FOSL1 was verified by PCR in MDA-MB-231. The cell proliferation ability and cell invasion ability of MDA-MB-231 were detected by MTT and Transwell assay, respectively. MSP was used to detect the methylation status of PRDM10 gene. The mRNA and protein expression levels of PRDM10 gene were detected by Q-PCR and Western-blot assay. Results MTT results showed that the optical density (OD) values were significantly lower in transfection group compared with those of control group at 24 h, 48 h and 72 h (all P<0.05), and the same as those compared with empty group at 48 h and 72 h (both P<0.05). Compared with empty group and control group, Transwell assays showed that the cell invasive abilities of MDA-MB-231 were decreased in transfection group (both P<0.05), and MSP assay showed that the methylation of PRDM10 gene was decreased in MDA-MB-231, and Q-PCR and Western-blot tests showed that the expressions of PRDM10 gene were increased in mRNA level and in protein level. Conclusion Silencing of FOSL1 gene inhibits the proliferation and invasion of MDA-MB-231 cells, which might be related to the demethylation of PRDM10 gene in the cells.

Objective To investigate the effect of the over-expressed CTCF on apoptosis factors Bax and Bcl-2 in human breast cancer cell line MDA-MB-231. Methods Reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expressions of CTCF,Bax and Bcl-2 in MDA-MB-231. The overexpression vector of CTCF/pEGFP-N1 was constructed. The overexpression plasmid CTCF/pEGFP-N1 and the empty vector plasmid pEGFP-N1 were transfected into breast cancer cell line MDA-MB-231 by lentivirus transfection, and the MDA-MB-231 cells were divided into CTCF group and control group. After successfully transfection of MDA-MB-231 identified by RT-PCR, real time quantitative PCR (Q-PCR) was used to detect the mRNA levels of Bax and Bcl-2 in MDA-MB-231 of the CTCF group and the control group. The protein levels of Bax and Bcl-2 were detected by Western blot assay and enzyme-linked immunosorbent assay (ELISA). Results The expression of CTCF was not found in MDA-MB-231, and expressions of Bax and Bcl-2 were found in MDA-MB-231. Results of Q-PCR showed that the mRNA levels of Bax were 4.63±1.08 and 2.27±0.16 in CTCF group and control group, respectively, and they were statistically significant (t=27.50, P<0.05). The mRNA levels of Bcl-2 were 1.39±0.14 and 3.56 ± 0.97 in CTCF group and control group, and there was significant difference between two groups(t=39.00, P<0.05). Results of Western blot assay showed that the protein level of Bax was higher in CTCF group compared with that of control group. The protein level of Bcl-2 was lower in CTCF group compared with that of control group. Results of ELISA showed that the protein levels of Bax were 15.25±2.17 and 6.24±1.78 in CTCF group and control group, respectively, and there was significant difference between the two groups (t=26.84, P<0.05). The protein levels of Bcl-2 were 4.59 ± 0.97 and 10.68 ± 1.93, and there was significant difference between the two groups (t=21.72, P<0.05). Conclusion The over-expressed CTCF can promote the expression of apoptotic factors and inhibit the expression of anti-apoptotic factors in breast cancer cells.

OBJECTIVETo investigate the effects of rhubarb powder on serum complement 3 (C3), complement 4 (C4), and hypersensitive C-reactive protein (hs-CRP) levels in patients with hypertensive intracerebral hemorrhage (HICH) after operation.

METHODSForty inpatients with HICH after operation were recruited from Department of Cerebral Surgery, Affiliated Hospital of Shaanxi College of Traditional Chinese Medicine from July 2009 to March 2010. They were randomly assigned to the treatment group (20 cases) and the control group (20 cases). From the 4th day after surgery, all patients received routine Western medical treatment. The rhubarb powder, 5-10 g dissolving in 40 mL warm water, was administered or nasally fed to those in the treatment group, 2 -3 times daily for 10 successive days. The contents of serum C3, C4, and hs-CRP were detected in the two groups on the 7th day and the 14th day after operation. The serum hs-CRP content was detected using latex particle enhanced immunoturbidimetric assay. The Scandinavia Stroke Scale (SSS) scores were recorded in the two groups.

RESULTSCompared with the same group on the 4th day after operation, the levels of serum C3 and C4 increased on the 7th day after operation, and SSS score increased on the 14th day after operation in the control group (P < 0.05). The contents of C4 and hs-CRP decreased, and the SSS score increased on the 14th day after operation in the treatment group (P < 0.05). Compared with the same group on the 7th day after operation, the contents of C4 and hs-CRP decreased and the SSS score increased on the 14th day after operation in the treatment group (P < 0.05). Compared with the control group at the same time points, the contents of C4 and C3 decreased on the 7th day after operation; the contents of C3, C4, and hs-CRP decreased, and SSS score increased in the treatment group on the 14th day after operation (P < 0.05).

CONCLUSIONThe rhubarb powder could significantly decrease the serum levels of C3, C4, and hs-CRP, and improve the curative effect in patients with HICH after operation.

Adult ; C-Reactive Protein ; metabolism ; Cerebral Hemorrhage ; blood ; Complement C3 ; metabolism ; Complement C4 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Male ; Middle Aged ; Postoperative Period ; Rheum ; chemistry

Objective To investigate effects of 5-Aza-2′-deoxycytidine (5-Aza-CdR) on proliferation of human breast cancer cell line Hs578T,and the methylation status of PRDM10 gene in vitro in this cell line.Methods The human breast cancer cell line Hs578T was cultured with 1,3 and 5 μmol/L DNA methylation inhibitor 5-Aza-CdR respectively, and untreated cells were used as control.Cell proliferation was detected by MTT assay.Methylation-Specific PCR(MSP)was used to detect the methylation status of PRDM10 gene. The mRNA and protein expression levels of PRDM10 gene were detected by RT-PCR and Western blot assay. Results MTT results showed that the higher the concentration of 5-Aza-CdR,and the longer the treatment time,the more significant inhibitory effect on the proliferation of Hs578T cells.Compared with the control group(0 μmol/L),the proliferation of Hs578T was significantly inhibited after the treatment for 72 h in the 1 μmol/L group, and for 48 h in the 3 μmol/L and 5 μmol/L groups (P<0.05). MSP results showed that the higher the concentration of 5-Aza-CdR,the more significant demethylation of PRDM10.Results of RT-PCR and Western blot showed that the higher the concentration of 5-Aza-CdR, the higher the expression levels of mRNA and protein in PRDM10 (P<0.05).Conclusion 5-Aza-CdR could inhibit the cell proliferation of Hs578T,which might be related to the demethylation of PRDM10 gene in the cells.