Carcinoma, Hepatocellular ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Interleukin-6 ; metabolism ; Liver Neoplasms ; metabolism ; pathology

Whether or not an abormal expression of IL-6 mRNA in PBMNCs from IDDM patientswas examined using a hihgly sensitive,specific and semiquantitative protocal,i.e.reverse tran-scription and polymerase chain reaction (RT-PCR).The relative levels of IL-6mRNA in PBM-NCs from 12 early IDDMpatients (8.20?3.85yr),29 newly diagnosed NIDDM patients(54.85?9.12yr)23 normal childrens (8.20?3.26yr) and 12 normal adults (31.92?11.22yr)weredetermined.Significantly high expresion levels of IL-6 mRNA were found in PBMNCs from pa-tients with IDDM (P

OBJECTIVETo investigate the expression of Oct4, Sox2, Nanog, SMO, beta-Catenin and Wnt5b mRNA in four hepatocellular carcinoma cell lines of SMMC-7721, Bel-7402, HepG2, MHCC-97 and normal hepatocellular cell line of L02, and to compare the response of these cell lines to all-trans retinoic acid.

METHODSRT-PCR was used to detect expression of Oct4, Sox2, Nanog, SMO, beta-Catenin and Wnt5b mRNA in four hepatocellular carcinoma cell lines and normal hepatocellular cell line. Real time-PCR was used to quantify the expression of the genes.

RESULTSThere are different levels of expression of the stem cell-related gene in hepatocellular carcinoma cell lines and control cell line (P less than 0.05). There are significant differences in HepG2 and L-02 for the response to all-trans retinoic acid (P less than 0.05).

CONCLUSIONSThe stem cell-related genes are differentially expressed in different hepatocellular carcinoma cell lines.

Carcinoma, Hepatocellular ; metabolism ; pathology ; DNA Methylation ; Gene Expression Regulation, Neoplastic ; Hep G2 Cells ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Nanog Homeobox Protein ; Octamer Transcription Factor-3 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; SOXB1 Transcription Factors ; genetics ; metabolism ; Signal Transduction ; Smoothened Receptor ; Stem Cells ; drug effects ; metabolism ; Tretinoin ; pharmacology ; Wnt Proteins ; genetics ; metabolism ; beta Catenin ; genetics ; metabolism

OBJECTIVEHypoxia/KCl injury model in the cultured neonatal rat cardiomyocytes (CMs) was established to investigate the protective effect of Lycium barbanun Glycopeptide (LbGp) on calcium overload.

METHODCultured neonatal rat CMs were divided into three groups, namely normal control, hypoxia groups and LbGp-treated group. CMs in LbGp-treated group and hypxia group were cultured in an incubator ventilated with 95% N2 and 5% CO2 with or without LbGP. CMs viability under hypoxia was measured by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide colorimetry (MTT). The intracellular free calcium concentration in cardiomyocytes was measured by laser confocal microscope with Fura-3/AM as a calcium indicator. The protective effects of LbGp on the CMs treated by KCl (60 mmol x L(-1)) was observed.

RESULTAs compared with normal controls, the degree of MTT metabolism was significantly reduced (P < 0.01) in hypoxic group and slightly reduced in LbGp (P < 0.05). Hypoxia-induced enhancement of intracellular calcium ([Ca2+]i) was attenuated by LbGp significantly (P < 0.01). Moreover, KCl-induced enhancement of [Ca2+]i was also reduced by LbGp at the doses of 25, 50, 100 microg x mL(-1) in a concentration-dependent manner.

CONCLUSIONThe result suggested that LbGp is able to increase the survival ratio and inhibit the enhancement of the intracellular free calcium concentration in cardiomyocytes induced by hypoxia and high potassium. One of the mechanisms is that LbGp acts on L-type calcium channels.

Animals ; Animals, Newborn ; Calcium ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Dose-Response Relationship, Drug ; Glycopeptides ; administration & dosage ; isolation & purification ; pharmacology ; Lycium ; chemistry ; Myocytes, Cardiac ; cytology ; metabolism ; Plants, Medicinal ; chemistry ; Potassium Chloride ; Rats ; Rats, Sprague-Dawley

Adult ; Aged ; Female ; Fluid Therapy ; methods ; Humans ; Intensive Care Units ; Male ; Middle Aged ; Pancreatitis ; mortality ; therapy ; Plasma ; Young Adult

In order to study the epidemiological characteristics of cytomegalovirus (CMV) infection in allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients by means of plasma real time quantitative polymerase chain reaction (RQ-PCR), 141 adult patients undergoing allo-HSCT between January 2008 and June 2010 were serially monitored by RQ-PCR for detecting CMV and guiding the preemptive therapy followed up to 180 days post-HSCT. The results showed that the incidence of CMV infection and CMV pneumonia was 81.5% and 2.9% respectively, which mainly occurred within 2 months post-HSCT. Single-therapy with ganciclovir (GCV) for 63 patients or foscarnet 6 patients was performed for preemptive therapy. The total efficacy was 87.8%, and the response patterns were different. CMV infection was more frequent in female patients (P = 0.044), and those with aGVHD (P = 0.043), using ATG or basiliximab in conditioning regimens (P = 0.049), as well as earlier in patients using ATG or basiliximab or those with aGVHD (P = 0.007; P = 0.000). The aGVHD, maximum load, positive times of CMV-DNA detection and therapy duration all correlated with the efficacy (P < 0.05). It is concluded that the incidence of CMV infection is still high after HSCT. Plasma RQ-PCR assay for CMV-DNA shows a strong correlation with the clinical outcome of CMV infection, which is useful and suitable for management of CMV infection in HSCT.
Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; Child ; Cytomegalovirus ; Cytomegalovirus Infections ; diagnosis ; drug therapy ; etiology ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Retrospective Studies ; Transplantation, Homologous ; Young Adult

OBJECTIVETo optimize the prenatal diagnosis platform by using domestically made fluorescence in situ hybridization(FISH) kit and to explore the clinical application of FISH to rapid prenatal diagnosis of a wide range of chromosomal abnormalities.

METHODSAmniotic fluid samples from 110 pregnant women were studied with the rapid prenatal diagnosis method of FISH and the conventional cell culture method of karyotyping, the results from both methods were compared.

RESULTSFour cases of trisomy 21, 1 case of trisomy 18, 58 cases of 46, XX, and 47 cases of 46, XY were detected by FISH in the 110 amniotic fluid samples. It is concordant with the results from conventional karyotype analysis. The concordance rate is 100%.

CONCLUSIONDomestically made FISH kit can be used to rapidly and accurately detect the most common chromosome aneuploidies by using less sample volume while the price is relatively low. FISH can be a reliable and rapid prenatal diagnostic tool as an adjunct to classical cytogenetic study. It can be used for rapid and accurate prenatal diagnosis of women with high risk of maternal serum screening.

Adult ; Amniocentesis ; Amniotic Fluid ; Aneuploidy ; Chromosome Aberrations ; Chromosomes, Human, Pair 18 ; genetics ; Down Syndrome ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Karyotyping ; methods ; Nucleic Acid Hybridization ; Pregnancy ; Prenatal Diagnosis ; methods ; Trisomy

OBJECTIVETo evaluate the efficacy and safety of selective serotonin re-uptake inhibitors (SSRIs) in the treatment of premature ejaculation (PE).

METHODSFrom MEDLINE (Jan, 1950-Mar, 2008), EMBASE (Jan, 1980-Mar, 2008), The Cochrane Library (Issue 1, 2008) and CNKI (Jan, 1979-Mar, 2008), we retrieved and screened the randomized controlled trials (RCT) and randomized crossover trials (RT) as well as various related data, published and unpublished, on the treatment of PE with SSRIs. The methodological quality of the included trials was evaluated by 2 reviewers. Meta-analyses were conducted with RevMan 5.0 on the homogeneous studies.

RESULTSTotally 22 studies on 4 291 patients were included. Meta-analyses showed that after treated with sertraline, fluoxetine, paroxetine, citalopram, dapoxetine and fluvoxamine, the WMD (95% CI) values of the changes in intravaginal ejaculatory latency time (IELT) were 2.63 (1.80, 3.46), 2.21 (1.50, 2.92), 4.31 (2.71, 5.91), 3.82 (3.39, 4.25), 1.57 (1.31, 1.84) and 0.01 (0.71, 0.73) respectively; the RR (95% CI) values of the sexual satisfaction rate of the patients were 1.65 (1.12, 2.43), 2.93 (0.50, 17.31), 3.08 (2.27, 4.17), 2.48 (1.99, 3.09) and 2.93 (2.36, 3.65), and those of their partners were 1.47 (0.98, 2.21), 2.88 (0.38, 21.77), 4.81 (3.15, 7.36), 5.38 (3.75, 7.72) and 2.91 (1.09, 7.78) respectively for sertraline, fluoxetine, paroxetine, citalopram and dapoxetine.

CONCLUSIONAll the known SSRIs but fluvoxamine could prolong IELT, and some could improve the sexual satisfaction of both the patients and their partners, but their adverse effects should be noted. The moderate possibility of selection bias and publication bias in the included studies might have a negative impact on the evidence intensity of our results. We expect more reliable evidence from more randomized controlled trials.

Humans ; MEDLINE ; Male ; Randomized Controlled Trials as Topic ; Serotonin Uptake Inhibitors ; adverse effects ; therapeutic use ; Sexual Dysfunction, Physiological ; drug therapy ; Treatment Outcome

OBJECTIVETo evaluate the endourological technique in the treatment of bilateral malignant ureteric obstruction.

METHODSThe data of 43 patients (totally, 70 cases) with bilateral malignant ureteric obstruction treated with endoluminal therapy were reviewed retrospectively. Of 70 cases, 38 were treated by retrograde double-J stenting, 24 by minimally invasive percutaneous nephrotomy (MPCN) and 8 by antegrade double-J stenting.

RESULTSAll patients were followed up for an average of 12 months. The retrograde double-J stenting, MPCN and antegrade double-J stenting was successfully performed in 50.0% (19/38), 100.0% (24/24) and 62.5% (5/8), respectively. Technical failures in placing retrograde double-J stent were too difficult to identify the ureteric orifice (13/38) or failing to cross the obstruction site because of severe extraluminal compression (6/38). Failure in placing antegrade double-J stent was due to severe extraluminal compression (3/8). Dislodgment of nephrostomy tubes (11/19) was the major factor which limited the application of MPCN.

CONCLUSIONIt is safe and effective to treat malignant ureteric obstruction with endourological technique, and suggested initially with retrograde double-J stenting. If malignant ureteric orifice occlusion or a severe extraluminal compression is showed in the imaging, MPCN or antegrade double-J stenting may be selected according to the site and the extent of obstruction.

Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Nephrostomy, Percutaneous ; methods ; Prostatic Neoplasms ; complications ; Retrospective Studies ; Stents ; Stomach Neoplasms ; complications ; Treatment Failure ; Treatment Outcome ; Ureteral Obstruction ; etiology ; surgery ; Urinary Bladder Neoplasms ; complications ; Uterine Cervical Neoplasms ; complications

OBJECTIVEThe prostate apoptosis response factor-4 (Par-4) gene was originally identified by differential screening for genes that are up-regulated when prostate cells are induced to undergo apoptosis. Par-4 was found to possess potent apoptotic activity in various cellular systems in response to numerous stimuli. The aim of this study was to explore the effects of small interfering RNA (siRNA) against Par-4 gene on the apoptosis of human bone marrow mesenchymal stem cells (hBMSCs) exposed to glutamate.

METHODSPrimary culture of hBMSCs was carried out and siRNAs targeted Par-4 gene (Par-4-SiRNA) were chemically synthesized. Eukaryocytic expression vector was built and were transfected into hBMSCs with liposome. After selecting with G418, the stable cell clones were treated with glutamate. The expression of Par-4 mRNA was determined by real-time PCR. The apoptosis of hBMSCs was quantified by flow cytometry. Western blotting was used to detect the protein levels of phosphorylated Akt1 (Thr308). Relative Caspase-3 activity was determined by colorimetric assay.

RESULTSThe Par-4-SiRNA-1 and Par-4-siRNA-2 could markedly down-regulate the mRNA levels of Par-4 gene in hBMSCs. With the transfections of Par-4-SiRNA-1 and Par-4-SiRNA-2, the levels of Par-4 mRNA were respectively decreased by 88% and 67%. Both Par-4-SiRNA-1 and Par-4-SiRNA-2 inhibited significantly the apoptosis of hBMSCs induced by glutamate, in which the percentages of apoptotic cells were respectively decreased to 38.80% +/- 3.97% (P < 0.01) and 45.49% +/- 4.32% (P < 0.01) from 60.30% +/- 6.82%. Western blot assays demonstrated that, glutamate down-regulated the expression of phosphorylated Akt1 proteins in hBMSCs (89.07 +/- 6.42 and 28.30 +/- 5.65, respectively, P < 0.01). However, Par-4-SiRNA-1 and Par-4-SiRNA-2 could markedly recover the down-regulation of Akt1 proteins induced by glutamate (63.56 +/- 6.75 and 45.59 +/- 4.88, respectively, P < 0.01). And the relative Caspase-3 activity which was enhanced by the treatment with glutamate (0.1428 +/- 0.0495 and 0.8616 +/- 0.1051, P < 0.01), was suppressed by Par-4-SiRNA-1 and Par-4-SiRNA-2 (0.8616 +/- 0.1051 and 0.6581 +/- 0.0555, respectively, P < 0.01).

CONCLUSIONSiRNA against Par-4 gene could inhibit the apoptosis of hBMSCs induced by glutamate, and its inhibitory effects may be mediated by the up-regulation of phosphorylated Akt1 and the suppression of the relative Caspase-3 activity.

Apoptosis ; genetics ; Apoptosis Regulatory Proteins ; genetics ; Bone Marrow Cells ; cytology ; metabolism ; Caspase 3 ; metabolism ; Cells, Cultured ; Gene Expression Regulation ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Small Interfering