AIM:To characterize the effect of estradiol on proliferation,differentiation and extracellular matrix(ECM) accumulation in stromal cells through regulation of BPH-1 paracrine.METHODS:BPH-1 cells were stimulated with different concentrations of estradiol.Conditioned media(CM) were harvested and their effects on stromal cell cultures were tested.Cell proliferation was determined by MTT assay.mRNA of smoothelin,fibronectin,collagen Ⅳ and transforming growth factor ?1(TGF-?1) were analyzed by real-time RT-PCR.Western blotting was used to determine smooth muscle myosin heavy chain(SMMHC).ELISA and radioimmunoassay were respectively used to measure fibronectin,TGF-?1 and collagen Ⅳ protein expressions.RESULTS:Estrodiol stimulated the expression and secretion of TGF-?1 in BPH-1 cells.The proliferation of stromal cells increased when they were cultured with CM harvested from estrogen treated BPH-1 cells.The mRNA levels of collagen Ⅳ and smoothelin increased in stromal cells treated with CM from BPH-1 cells.The results of radioimmunoassay also showed that the collagen Ⅳ protein level up-regulated in the supernatants and cell extracts of CM-treated stromal cells.A neutralizing antibody to TGF-?1 inhibited the stimulation of collagen Ⅳ and SMMHC by BPH-1 CM.The expression of fibronectin was only marginally changed in stromal cells cultured in the presence of BPH-1 CM.CONCLUSION:The BPH-1 cells increase ECM accumulation and differentiation of stromal cells through TGF-?1.Estradiol stimulate differentiation of stromal cells by induction of TGF-?1 expression.Estradiol stimulate proliferation by influencing the factors secreted from prostatic epithelial cells.

Objective To evaluate the efficacy and feasibility of laparoscopie intervention for congenital obstructive megaureter in children. Methods Eleven children with congenital obstructive megaureter(left in 4,right in 7)underwent laparoseopie ureteroplasty.One had congenital ureter oririce stenosis,9 had been diagnosed as simple congenital ureter orifice stricture,1 had recurrent ureter orifice stricture after open ureterovesical reimplantation.B-ultrasound and IVU showed severe hydronephrosis in 7 cases and moderate in 4. Results The operation was successful in all cases and none had urine leakage.The mean operating time was 103.0±35.3 min(range 70-190 min).The mean blood loss was 18.0±9.5 ml(range 10-40 ml)and the mean postoperative hospital stay was 8.0±1.4 d(range 7-10 days).The double J stent was removed 6 weeks after operation.The patients were followed up for 3-24 months(mean,6 months).Cystography showed no reflux in all cases during follow-up. Conclusion Laparoscopical ureteroplasty could be a minimal invasive,less suffering technique for the treatment of congenital obstructive megaureter in children.

BACKGROUND:Gemcitabine hydrochloride is a water-soluble anticancer drug that induces apoptosis in tumor cells, but it has an excessive release in vivo. OBJECTIVE:To evaluate the effect of poly(lactic acid) (PLA) electrospun fiber membranes carrying gemcitabine hydrochloride on the growth of human osteosarcoma cell lines MG-63. METHODS:PLA electrospun fiber members with (experimental) or without (control) gemcitabine hydrochloride were fabricated and characterized. Two kinds of fiber membranes were immersed in low-glucose DMEM medium, and the supernatants were collected in the two groups at 3, 5, 7 days, respectively. Passage 5 human osteosarcoma cell lines MG-63 were inoculated into 96-well plates containing low-glucose DEME with 15％ fetal bovine serum, and divided into seven groups. Groups 1-3 were cultured in the experimental supernatants of 3, 5, 7 culture days, and groups 4-6 were cultured in the control supernatants of 3, 5, 7 culture days, respectively. The remaining group acted as the negative control with no supernatant. Thereafter, cell counting kit-8 was used to detect cell proliferation, and RT-PCR was used to measure expression of Bcl-2 and Bax at 3 days of culture. RESULTS AND CONCLUSION:(1) No obvious particle was found on the smooth and even surface of the fiber members in the experimental and control groups. There was no significant difference in fiber diameter, contact angle and tensile strength between the two kinds of fiber membranes. (2) The results of cell counting kit-8 showed that compared with the negative control group, the supernatant released from the control group had no effect on the MG-63 proliferation at different time points, while the supernatant released from the experimental group could inhibit the MG-63 proliferation at different time points (P<0.05), and the inhibitory effect became more and more obvious with the prolongation of release time. (3) RT-PCR findings showed that compared with the control group, the supernatant released from the experimental group could increase Bax mRNA expression and decrease Bcl-2 mRNA expression at the same time point. To conclude, the PLA electrospun fiber membranes carrying gemcitabine hydrochloride can sustainably inhibit MG-63 proliferation and promote cell apoptosis.

OBJECTIVETo study the function of the SP3111 protein in fertilization and early embryo development through in vitro fertilization (IVF) experiments following anti-SP111 antibody (Ab2438) blocking.

METHODSSperm samples collected from male mice were divided into an experimental, a blank control and a negative control group before IVF. The sperm of the experimental group was incubated with Ab2438 for 1 h followed by IVF and observed for the rates of fertilization and embryo fragmentation at 2, 4, 6, 8 and 22 h. Then the fertilized eggs were incubated with Ab2438, and the rates of fertilization embryo fragmentation were observed at 22 h.

RESULTSAfter the sperm was incubated with Ab2438, the incidences of embryo fragmentation were 5.26, 8.77, 23.25, 43.42 and 59.21% at 2, 4, 6, 8 and 22 h, respectively, with significant differences from the control groups (P < 0.01). After 22 h Ab2438 incubation of the fertilized eggs, the rates of normal and fragmented embryos of the experimental group were 23.64 and 63.64%, respectively, significantly different from those of the control groups (P < 0.01).

CONCLUSIONAnti-SP3111 antibodies remarkably affected fertilization and early embryo development in mice. The SP3111 protein may be a signal molecule and plays a role in fertilization and early embryo development together with other proteins. Further studies on the function of the SP3111 protein in reproduction may offer a new insight into the molecular mechanism of infertility.

Animals ; Antibodies ; immunology ; Embryonic Development ; Female ; Fertilization in Vitro ; Male ; Membrane Proteins ; metabolism ; Mice ; Mice, Inbred ICR ; Spermatozoa ; immunology

OBJECTIVETo determine the effects of Chinese herbal monomers such as baicalin, berberine, and matrine on the androgen receptor (AR) mRNA expression in SZ95 sebocytes in vitro and to explore the possible mechanism of using traditional Chinese medicines to treat acne.

METHODSSZ95 sebocytes were cultured and then treated with berberine, baicalin, matrine, and 13-cis-retinoic acid for 24 hours. Reverse transcription polymerase chain reaction was applied to detect the changes of AR.

RESULTAR mRNA was downregulated by 13-cis-retinoic acid of 1 x 10(-5) mol/L and 1 x 10(-6) mol/L, and by baicalin of 1 x 10(-4) mol/L (P < 0.05).

CONCLUSION13-cis-retinoic acid and baicalin may exert antiandrogenitic action by inhibiting AR mRNA expression in human sebocytes.

Androgen Antagonists ; pharmacology ; Cell Line ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Flavonoids ; pharmacology ; Humans ; RNA, Messenger ; biosynthesis ; Receptors, Androgen ; biosynthesis ; genetics ; Skin ; cytology

OBJECTIVETo investigate the effect of ionizing radiation on the expression of p16, CyclinD1, and CDK4 in mouse thymocytes and splenocytes.

METHODSFluorescent staining and flow cytometry analysis were employed for the measurement of protein expression.

RESULTSIn time course experiments, it was found that the expression of p16 protein was significantly increased at 8, 24, and 48 h for thymocytes (P < 0.05, P < 0.01, and P < 0.05, respectively) and at 24 h for splenocytes (P < 0.05) after whole body irradiation (WBI) with 2.0 Gy X-rays. However, the expression of CDK4 protein was significantly decreased from 8 h to 24 h for thymocytes (P < 0.05-P < 0.01) and from 8 h to 72 h for splenocytes (P < 0.05-P < 0.01). In dose effect experiments, it was found that the expression of p16 protein in thymocytes and splenocytes was significantly increased at 24 h after WBI with 1.0, 2.0, and 4.0 Gy (P < 0.05-P < 0.01), whereas the expression of CDK4 protein was significantly decreased with 2.0 Gy for thymocytes (P < 0.05) and 0.5-6.0 Gy for splenocytes (P < 0.05-P < 0.01). Results also showed that the expression of CyclinD1 protein decreased markedly in both thymocytes and splenocytes after exposure.

CONCLUSIONThe results indicate that the expression of p16 protein in thymocytes and splenocytes can be induced by ionizing radiation, and the p16-CyclinD1/CDK4 pathway may play an important role for G1 arrest of thymocytes induced by X-rays.

Animals ; Cyclin D1 ; biosynthesis ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; Cyclin-Dependent Kinases ; biosynthesis ; Dose-Response Relationship, Radiation ; Flow Cytometry ; Male ; Mice ; Mice, Inbred Strains ; Proto-Oncogene Proteins ; Radiation Dosage ; Spleen ; cytology ; metabolism ; radiation effects ; Thymus Gland ; cytology ; metabolism ; radiation effects ; X-Rays

BACKGROUNDModern research has provided new insights into the biological mechanisms of noise-induced hearing loss, and a number of studies showed the appearance of increased reactive oxygen species (ROS) and reactive nitrogen species (RNS) during and after noise exposure. This study was designed to investigate the noise exposure induced nitrotyrosine change and the mechanism of outer hair cells death in guinea pig cochlea.

METHODThirty guinea pigs were used in this study. The experimental animals were either exposed for 4 hours per day to broadband noise at 122 dB SPL (A-weighted) for 2 consecutive days or perfused cochleae with 5 mg/ml of the SIN1 solutions, an exogenous NO and superoxide donor, for 30 minutes. Then the cochleae of the animals were dissected. Propidium iodide (PI), a DNA intercalating fluorescent probe, was used to trace morphological changes in OHC nuclei. The distribution of nitrotyrosine (NT) in the organ of Corti and the cochlear lateral wall tissue from the guinea pigs were examined using fluorescence immunohistochemistry method. Whole mounts of organ of Corti were prepared. Morphological and fluorescent changes were examined under a confocal microscope.

RESULTSEither after noise exposure or after SIN1 perfusion, outer hair cells (OHCs) death with characteristics of both apoptotic and necrotic degradation appeared. Nitrotyrosine immunolabeling could be observed in the OHCs from the control animals. After noise exposure, NT immunostaining became much greater than the control animals in OHCs. The apoptotic OHC has significant increase of nitrotyrosine in and around the nucleus following noise exposure. In the normal later wall of cochleae, relatively weak nitrotyrosine immunolabeling could be observed. After noise exposure, nitrotyrosine immunoactivity became stronger in stria vascularis.

CONCLUSIONNoise exposure induced increase of nitrotyrosine production is associated with OHCs death suggesting reactive nitrogen species participation in the cochlear pathophysiology of noise-induced hearing loss.

Animals ; Cell Death ; Cochlea ; chemistry ; pathology ; Female ; Guinea Pigs ; Hair Cells, Auditory, Outer ; pathology ; Immunohistochemistry ; Male ; Noise ; adverse effects ; Organ of Corti ; chemistry ; pathology ; Tyrosine ; analogs & derivatives ; analysis

OBJECTIVETo investigate the pathway and mechanism of noise exposure induced out hair cells (OHC) apoptosis.

METHODSThe cochleae of control and noise exposure group were dissected. The activity of caspase 3, an important mediator of apoptosis, in OHC, was examined with carboxyfluorescein-labeled fluoromethyl ketone (FMK)-peptide inhibitors. The apoptosis inducing factor (AIF) translocation from mitochondria in OHC were further examined by immunohistology method. The nuclei were labeled with PI and the mitochondrion was labeled with Mito-tracker. Whole mount organ of Corti was prepared. Morphological and fluorescent change was observed use confocal microscope.

RESULTSIn the normal OHC, AIF is distributed where the mitochondria were located and no activated caspase 3 was observed. After the animals exposed to broadband noise at 122 dB in 4 h/day for 2 days, both apoptosis and necrosis were appeared in OHC. AIF translocated from mitochondrion to nuclei in apoptotic and necrotic OHC following noise exposure. The noise exposure triggered activation of caspase 3 in apoptic hair cells. But no caspase 3 activation appeared in necrotic OHC.

CONCLUSIONSThese findings indicated that the caspase-dependent pathway is an important pathway in noise exposure induced apoptosis. And AIF also involves OHC death pathway following noise exposure.

Animals ; Apoptosis ; Apoptosis Inducing Factor ; metabolism ; Caspase 3 ; metabolism ; Female ; Guinea Pigs ; Hair Cells, Auditory, Outer ; metabolism ; pathology ; Male ; Noise

The human basic Krüppel-like factor (hBKLF) is a newly cloned human transcription factor from the cDNA library of fetal liver. It belongs to the Krüppel-like transcription factor family. Previous expression study showed that it is a hematopoietic related factor. This study was aimed to investigate the effect of hBKLF on cell proliferation, differentiation and hemoglobin synthesis by using K562 cell line as model. The sense and antisense expression plasmids of hBKLF were constructed, and transfected into K562 cells by lipofectamine. After G418 selection for 4 weeks, the cell line with stable expression of the gene was obtained. Then the hBKLF expression level, proliferation ability, colony formation and hemoglobin production were detected by RT-PCR and Western blot, MTT method, methyl cellulose semisolid culture method and benzidine test respectively. The morphologic change of cell was observed with inverted microscope. The results showed that the sense plasmid could increase hBKLF level and antisense plasmid could decrease hBKLF expression. When hBKLF level was down-regulated, K562 cells could proliferate more quickly and synthesize more hemoglobin. But there were no differences in colony formation ability and no apparent morphologic change. It is concluded that hBKLF can inhibit hematopoietic cell proliferation and hemoglobin synthesis. It is suggested that hBKLF plays an important role in the proliferation and differentiation of hematopoietic cells.
Animals ; COS Cells ; Cell Differentiation ; physiology ; Cell Proliferation ; drug effects ; Cell Transformation, Neoplastic ; drug effects ; Cercopithecus aethiops ; Hemoglobins ; biosynthesis ; Humans ; K562 Cells ; Kruppel-Like Transcription Factors ; biosynthesis ; genetics ; pharmacology ; Transcription Factors ; biosynthesis ; genetics ; Transfection

OBJECTIVETo review the research progress on Type IV secretion system (T4SS) in Helicobacter pylori.

DATA SOURCESThe data used in this review were identified by searching of PUBMED (1995 - 2007) online resources using the key terms 'Type IV secretion system' and 'Helicobacter pylori'.

STUDY SELECTIONMainly original articles and critical reviews written by major pioneer investigators of this field were selected.

RESULTSThe research progress on T4SS in Helicobacter pylori was summarized. The structure and function was discussed.

CONCLUSIONST4SS is not only involved in toxin secretion and injection of virulence factors into eukaryotic host target cells, but also involved in horizontal DNA transfer to other bacteria and eukaryotic cells, through DNA uptake from or release into the extracellular milieu. It provides a new insight into the pathogenicity of Helicobacter pylori and a novel target for antimicrobials development. However, many challenges remain for us in understanding the biological role of T4SS in Helicobacter pylori.

Bacterial Proteins ; metabolism ; DNA-Binding Proteins ; Gene Transfer, Horizontal ; Helicobacter pylori ; genetics ; metabolism ; pathogenicity ; Multigene Family