OBJECTIVETo investigate the protective effect of hypothermia combined with dexamethasone on spermatogenesis and the expression of intercellular adhesion molecule 1 (ICAM1) after testicular torsion-detorsion.

METHODSWe made unilateral testicular torsion models in 100 pubertal male Sprague-Dawley rats by 720 degree torsion of the left testis and then randomly divided them into four groups of equal number to be treated with normal temperature + physiological saline (group A), hypothermia + physiological saline (group B), normal temperature + dexamethasone (group C), and hypothermia + dexamethasone (group D). After 48 hours, we collected the testes, observed pathological changes of the testicular tissue by HE staining under the light microscope, detected the apoptosis of spermatogenic cells by TUNEL, and determined the expression of ICAM1 by Western blot.

RESULTSHE staining showed different degrees of testicular tissue injury in the four groups of rats, most obvious in group A, but mild in the other three. The ICAM1 protein expression was significantly higher in group A (0.68 +/-0. 03) than in B (0. 49 +/- 0. 06, P <0. 05) , C (0. 46 +/- 0. 09, P < 0.05) , and D (0.17 +/- 0.08, P <0.01). The nuclei were deep brown or brown. Lots of apoptotic spermatogenic cells were seen in the torsion testis of group A, with a significantly higher apoptosis index ( [33. 13 +/- 3.21 ]%) than in B ( [ 17. 12 +/-5.23 ]%, P < 0.05), C ([14.13 +/- 2.03]%, P <0.05), and D ([9.05 +/- 1.03]%, P <0.01).

CONCLUSIONHypothermia combined with dexamethasone can protect the testis from injury as well as the reproductive function of the testis after testicular torsion-detorsion and reduce the expression of ICAM1.

Animals ; Dexamethasone ; pharmacology ; Disease Models, Animal ; Hypothermia, Induced ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Spermatic Cord Torsion ; metabolism ; physiopathology ; Spermatogenesis ; drug effects

OBJECTIVETo investigate the effects of crypotanshinone (CPT) on the proliferation and apoptosis of DU145 prostate cancer cells as well as on the metadherin expression and the downstream PI3K/AKT signaling pathway in the DU145 cells.

METHODSWe treated DU145 prostate cancer cells with different concentrations of CPT for 24, 48, and 72 hours followed by evaluation of the proliferation and apoptosis of the cells by MTT assay and TUNEL, respectively. We determined the expressions of metadherin protein and mRNA in the DU145 cells by Western blot and RT-PCR respectively at different time points after CPT treatment. We also detected the expressions of the proteins metadherin, AKT, p-AKT, and Bcl-2 in the CPT-treated DU145 cells at 48 hours.

RESULTSCPT significantly inhibited the proliferation of the DU145 cells in a dose- and time-dependent manner (P < 0.05). After treatment with 10 µmol/L CPT for 24, 48, and 72 hours, the apoptosis rates of the DU145 cells were (29.42 ± 4.51), (55.07 ± 5.67) and (70.84 ± 4.66)%, respectively, significantly higher than (3.1 ± 2.48)% in the control group (P < 0.05). The expression of metadherin was remarkably downregulated at the transcription and translation levels (P < 0.05) and the expressions of the AKT signaling pathway and the Bcl-2 protein were markedly inhibited in the DU145 cells after treated with 10 µmol/L CPT for 48 hours (P < 0.05).

CONCLUSIONCPT can inhibit the proliferation and induce the apoptosis of DU145 prostate cancer cells, which may be associated with its suppression of the downstream PI3K/AKT signaling pathway by reducing the expression of metadherin in the DU145 cells.

Apoptosis ; drug effects ; Cell Adhesion Molecules ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diterpenes, Abietane ; pharmacology ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Humans ; In Situ Nick-End Labeling ; Male ; Neoplasm Proteins ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-akt ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; Signal Transduction ; drug effects ; Time Factors

To investigate the influence of HIF-1alpha overexpression on the differentiation of endothelial progenitor cells (EPCs) ex vivo, EPCs were isolated from human peripheral blood by density gradient centrifugation, overexpressed HIF-1alpha was transfected to EPCs by electroporation; HIF1alpha, HIF1beta, vascular endothelial growth factor (VEGF) mRNA level were measured with RT-PCR; HIF-1alpha protein was detected with immunohistochemistry in a time course. CD31(+) cells were measured with flow cytometry. Cell morphology was observed after transfection. The results showed that the transfection efficiency of HIF-1alpha to EPCs was about 20%. HIF-1alpha and its controlled target gene VEGF were markedly induced by HIF-1alpha vector (P < 0.05). HIF1beta had its same level as it before interference (P > 0.05). HIF-1alpha protein was induced by HIF-1alpha transfection after 12 hours but was undetectable at 24 hours. After 7 - 14 days cultured in 21% oxygen pressure, fluorescence-trace experiments revealed that CD31 + EPCs/EC could be generated more efficiently from overexpressed HIF-1alpha than that from pEGFP transfected group (P > 0.05). EPC morphology was observed by light microscopy. HIF-1alpha-transfected cells under normoxia sprouted more rapidly from the EPC colonies than the untransfected cells or cells transfected with an GFP vector, which essentially maintained the original colony formation. HIF-1alpha transfected cells took on an array-like arrangement rather than random dispersal, suggesting that they were in an advanced state of differentiation. It is concluded that the utility of overexpression of HIF-1alpha can induce target genes which have influence on cell differentiation. HIF-1alpha transfection was found to give a prospected way to do the insight research on ischemic treatment in vivo.
Cell Differentiation ; drug effects ; Endothelial Cells ; cytology ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; biosynthesis ; genetics ; Plasmids ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Stem Cells ; cytology ; Transfection ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

OBJECTIVETo investigate the protective effects of hypothermia combined with dexamethasone on the testis of rats after testicular torsion reduction and on the expression of eNOS and apoptosis of spermatogenic cells.

METHODSWe made unilateral testicular torsion models in 80 adolescent male Sprague-Dawley rats by 720 degrees torsion of the left testis, and then randomly divided them into four groups of equal number to be treated with normal temperature + physiological saline (group A), hypothermia + physiological saline (group B), hypothermia + dexamethasone (group C), and normal temperature + dexamethasone (group D). After 48 hours, we collected the testes, observed pathological changes of the testicular tissue by HE staining under the light microscope, determined the expression of eNOS by immunohistochemistry, and detected the apoptosis of spermatogenic cells by TUNEL.

RESULTSHE staining showed different degrees of testicular tissue injury in all the four groups of rats, most obvious in group A, while protective effect was observed in the other three groups. Immunohistochemistry revealed significantly more positive cells and higher positive staining intensity in the torsion (left) testis in group A than in B (P < 0.05), C (P < 0.01) and D (P < 0.01). The nuclei were deep brown or brown. Lots of apoptotic spermatogenic cells were seen in the torsion testis of group A, with a significantly higher apoptosis index (31.12 +/- 4.68) than in B (16.58 +/- 6.22) (P < 0.05), C (8.60 +/- 1.15) (P < 0.01) and D (13.52 +/- 3.06) (P < 0.01).

CONCLUSIONIschemia-reperfusion injury after testicular torsion reduction can increase the apoptosis of spermatogenic cells and decrease testicular reproductivity. Hypothermia combined with dexamethasone can protect the testis from injury as well as the reproductive function of the testis after testicular torsion reduction.

Animals ; Apoptosis ; Dexamethasone ; pharmacology ; Germ Cells ; cytology ; metabolism ; Hypothermia, Induced ; Male ; Nitric Oxide Synthase Type III ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spermatic Cord Torsion ; metabolism ; pathology ; Spermatogenesis ; Testis ; metabolism ; pathology

The aim of this study is to establish an HPLC method for simultaneous determinations of mifepristone and its metabolites, mono-demethylated mifepristone, di-demethylated mifepristone and C-hydroxylated mifepristone in plasma and to evaluate the pharmacokinetic characteristics of mifepristone tablet. Twenty healthy female Chinese subjects were recruited and a series of blood samples were collected before and after 0.25, 0.5, 1.0, 1.5, 2.0, 4.0, 8.0, 12.0, 24.0, 48.0, 72.0 and 96.0 hours administration by a single oral dose of 75 mg mifepristone tablet. Mifepristone and its three metabolites were extracted from plasma using ethyl acetate and determined by high performance liquid chromatography. The main pharmacokinetic parameters of mifepristone and its metabolites, including Cmax, tmax, MRT, t(1/2), V, CL, AUC(0-96 h) and AUC(0-infinity), were calculated by Drug and Statistical Software Version 2.0. The simple, accurate and stable method allows the sensitive determinations of mifepristone and its metabolites in human plasma up to 4 days after oral administration of 75 mg mifepristone tablet and the clinical applications of their pharmacokinetic studies.
Administration, Oral ; Area Under Curve ; Asian Continental Ancestry Group ; Biological Availability ; Chromatography, High Pressure Liquid ; methods ; Female ; Humans ; Mifepristone ; administration & dosage ; metabolism ; pharmacokinetics ; Tablets

Objective To evaluate the epidemiological effects of vaccine immunization program related to A(H1N1)influenza in the middle school students.Methods Non-randomized clinical trial was designed to assess the A(H1N1)influenza vaccine on its efficacy.14883 students from 8 middle schools in Zhejiang province were recruited and classified into vaccinated or control groups,based on the status of immunization with A(H1N1)influenza vaccine.All subjects were followed up through one epidemic period(6 months)and the incidence rates of influenza-like illnesses,A(H1N1)influenza,and seasonal influenza in these two groups were compared to evaluate the efficacy of the vaccine.Results There were 6334 subjects in the vaccinated group and 8549 in the control group.7441.75 person-years were followed from these two groups.The incidence rate of A (H1N1)influenza in vaccinated group was 1.64‰ per person-year,lower than that of the control group.The rate difference(RD)was-1.64‰ per person-year(95% confidence interval value from-3.04‰ to-0.23‰ per person-year),and the difference was significant(P=0.010).The incidence rate of influenza-like illnesses in vaccinated group was 21.47‰ per person-year,lower than that of the control group(22.69‰ per person-year)and the diffefence was not significant(P>0.05).The incidence rate of B influenza in vaccinated group was 6.63‰ per person-year,higher than that of control group(7.02‰ per person-year)but the difference was not significant(P>0.05).Conclusion This vaccine demonstrated a good epidemiological effect against the A(H1N1)influenza virus infection,observed through a student-immunization program.The cross-protection effect against the influenza-like illnesses and other seasonal influenzas was not noticed in this study.