Objective of this study was to investigate the changes of cyclooxygenase-2 expression and mitochondrial membrane potential in apoptotic NB4 cells induced by arsenic trioxide (As(2)O(3)). The morphological changes in apoptosis process of NB4 cells treated by arsenic trioxide were observed under immunofluorescence microscope and DNA electrophoresis method, and the apoptosis rate of NB4 cells and the variations of mitochondrial membrane potential were detected by flow cytometry. Furthermore, the variations of expression level of cyclooxygenase-2 protein were analyzed by using Western blot method. The results indicated that after NB4 cells were treated with 2 µmol/L As(2)O(3) for 48 hours, some variations of NB4 cells were observed, such as pyknosis, chromatin segmentation, even fragmentation. Meanwhile, the typical DNA Ladder phenomenon was observed. The apoptosis rate of NB4 cells treated with 3 µmol/L As(2)O(3) for 48 hours was 33.34%, Furthermore the apoptosis rate of NB4 cells was enhanced along with the increase of concentration of As(2)O(3). After NB4 cells were treated with 0.5, 1, 2, 4 and 8 µmol/L As(2)O(3) for 48 hours, the mitochondrial membrane potential decreased by 12.8%, 21.6%, 66.9%, 83.7% and 83.8% respectively. The Western blot detection results showed that the expression level of cyclooxygenase-2 protein in NB4 cells was lower than that in control cells and decreased along with the rise of As(2)O(3) concentration, then the negative dose-dependent manner was observed between these 2 groups. It is concluded that As(2)O(3) can effectively induce NB4 cell apoptosis, and the dose-dependent manner existed in certain extent of concentrations. The decrease of mitochondrial membrane potential may be related with NB4 cell apoptosis induced by As(2)O(3). Cyclooxygenase-2 participates in the process of NB4 cell apoptosis induced by As(2)O(3).
Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Cell Line, Tumor ; Cyclooxygenase 2 ; metabolism ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; Membrane Potential, Mitochondrial ; Oxides ; pharmacology

Objective To clarify the clinical features,therapeutic strategy and prognosis of lipid storage myopathy (LSM).Methods The clinical data and therapeutic effects of 42 LSM patients were summarized retrospectively.All patients were followed up to evaluate their prognosis.Results Data of short-term therapeutic results of all the 42 patients were available.Thirty-three cases were placed in low- doses prednisone and 9 cases in riboflavin.All patients showed marked and quick improvement of symptoms within one month.Among thirty-two patients followed up for more than one year,26 cases had a full recovery and 6 remained to have intolerance to heavy exercise.Thirteen patients had relapses of muscle weakness in various degrees and most of which were induced by exertion,exposure to coldness and upper respiratory tract infection.In 5 patients the symptoms were recurred for more than one time.Among 13 cases with relapses, 7 had family history.Conclusions Our data suggest that LSM is a treatable disease and well responsive to low-doses prednisone.The disease tends to recur,especially in patients with family history.Glutaric aciduria type Ⅱ should be considered in LSM patients who are responsive well to riboflavin,indicating drug therapeutic strategy for LSM should be based on the etiology of the disease.

Objective To observe the sequence of fat deposit and its relationship with insulin resistance in SD rats fed by high fat diet.Methods Normal 8-week-old male SD rats were randomly divided into normal chow (NC,n=40)and high fat diet(HF,n=40)groups.Triglyceride(TG)in serum,liver and muscle were measured;glucose infusion rate(GIR)and the mRNA level of genes related to lipid metabolism in liver and muscle were determined in different stages.GIR was detected by eugiyeemic-hyperinsulinemic clamp for evaluating the insulin sensitivity.Gene expression was determined by real-time PCR.Results(1)As compared with NC group,serum TG was not increased after high fat feeding for4 and 8 weeks,it began to increase after 12 weeks [0.52(0.15-1.00) mmol/L vs O.31(0.09-0.53)retool/L, P0.05)in skeletal muscle.After 8 weeks,the expression of ACC1 in liver in HF group was increased by 20.6%,CPT-1 was decreased by 27.1%(P

Objective To determine the expression of membrane-bound B lymphocyte stimulator (BLyS) protein and its mRNA in vitro of peripheral blood mononuclear cells (PBMCs) from individuals with systemic lupus erythematosus (SLE),and to investigate the role of interferon-?(IFN-?) on the expression of BLyS.Methods PBMCs were obtained from 25 SLE patients (mean age of 31+14) and 20 healthy volunteers (mean age of 28?10).They were randomized into IFN-?(5 ng/ml) group and control group.PBMCs were col- lected at 0,6,12 and 24 h for BLyS mRNA assessment using semi-quantitative reverse transcription-PCR (RT-PCR).PBMCs were also collected at 72 h for membrane-bound BLyS protein detection using flow cy- tometry (FACS) and direct immunofluorescence.Results①The expression of BLyS mRNA and membrane- bound protein in PBMCs was significantly higher in individuals with SLE compared with healthy controls (P＜0.05);②IFN-?enhanced BLyS mRNA expression in PBMCs in both healthy controls and SLE patients,with the greatest effect at 6 h (stimulated vs unstimulated,0.42?0.19 vs 0.25?0.14,P＜0.01;0.59?0.28 vs 0.44?0.21,P＜0.01 );③IFN-?also increased the expression of membrane-bound BLyS protein in both healthy con- trols and individuals with SLE (FACs,mean fluorescence intensity,4.5+3.0 vs 3.7~2.6,P

The mechanisms involved in virus-induced severe hepatitis have not been fully elucidated.In this study,we investigated the role of gamma delta T cell receptors (γδ) T cells in the pathogenesis of fulminant viral hepatitis (FVH) induced by murine hepatitis virus strain 3 (MHV-3).The model of FVH was established by intraperitoneal injection of MHV-3 into Balb/cJ mice.The survival days of mice,and the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were examined.The proportions ofγδ T cells in blood,spleen and liver,and cytokines secreted by hepatic γδ T cells were analyzed by flow cytometry.The function of hepatic γδ T cells was examined by cytotoxicity assay.Balb/cJ mice died in 3 to 6 days post MHV-3 infection,with severe hepatic necrosis and significant augmentation of serum ALT and AST levels.The proportions of γδ T ceils in blood,spleen and liver were significantly increased post MHV-3 infection,while those of the early activating molecule CD69-expressing γδ T cells and productions of cytokines tumor necrosis factor-alpha (TNF-α) and interferon-γ (IFN-γ) increased remarkably in the liver.These highly activated liver γδ T cells were cytotoxic to MHV-3-infected hepatocytes in vitro and this effect of liver γδ T cells against hepatocytes might involve the TNF-α and IFN-γ pathway.These results demonstrated that γδ T cells might contribute to the pathogenesis ofMHV-3-induced FVH through the effector cytokines TNF-α and IFN-γ.

AIMTo investigate the anti-inflammatory mechanism of esculentoside A (EsA) and to observe the effects of EsA on cellular adhesion between human umbilical vein endothelial cell (VEC304) and human neutrophil and to further observe the mRNA expression of intercellular adhesion molecule-1 (ICAM-1) and cluster of differentiation 18(CD18).

METHODSThe hemocyte counting method was used for assaying the adhesion rate between VEC304 and neutrophil. The RT-PCR method was used for measuring the mRNA expression of ICAM-1 and CD18.

RESULTSThe adhesion rate between VEC304 and neutrophil was increased with treatment of lipopolysaccharide(LPS). EsA (3 - 12 x 10(-6) mumol.L-1) was shown to inhibit the high cellular adhesion induced by LPS. A further investigation of adhesion molecules mRNA expression was undertaken using semi-quantitative reverse transcribed polymerase chain reaction (RT-PCR). The results of RT-PCR from VEC304 and human neutrophil treating with LPS showed that ICAM-1 and CD18 mRNA expressions were higher than those of normal cells, while this increased expression of ICAM-1 and CD18 mRNA was remarkably attenuated by the addition of EsA.

CONCLUSIONEsA was found to inhibit the increased adhesion rate induced by LPS. Moreover, LPS induced high expression of ICAM-1 and CD18 was inhibited with treatment of EsA. It might be involved in the mechanisms of anti-inflammation of EsA.

Adult ; CD18 Antigens ; biosynthesis ; genetics ; Cell Adhesion ; drug effects ; Cell Line ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Endothelial Cells ; metabolism ; physiology ; Humans ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Neutrophils ; metabolism ; physiology ; Oleanolic Acid ; analogs & derivatives ; isolation & purification ; pharmacology ; Phytolacca ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Saponins ; isolation & purification ; pharmacology ; Umbilical Veins ; cytology

Endothelin (ET) is a peptide released by vascular endothelial cells. Except for the potent vasoconstrictor function it plays an important physiological role in tissue differentiation and development, cell proliferation and hormone production. Investigation of the role of ET axis in a variety of tumors such as prostatic, cervical, breast carcinoma has provided evidences of its importance in cancer, recently. Data suggest that multiple functions of the ET axis have associations with mitogenesis, apoptosis inhibition, angiogenesis, and activation of proto-oncogene. The ET axis relates to invasiveness, osteoblast function, and metastatic cancer pain in advanced prostate cancer.
Animals ; Apoptosis ; Endothelins ; physiology ; Humans ; Male ; Mice ; Prostatic Neoplasms ; pathology ; physiopathology ; Receptors, Endothelin ; physiology

OBJECTIVETo investigate the effects of diet-induced obesity on the developmental process of testes in pubertal rats.

METHODSEighty 21-day-old male SD rats were randomly divided into a control group (n=32) and an experiment group (n=48), and fed respectively on a normal diet and a high-fat diet. And changes in the body weight, Lee's index, testis weight and epididymis weight were measured at the end of the 3rd, 4th, 5th and 6th week after the treatment, that is, when the rats were 6, 7, 8 and 9 weeks old. The concentrations of testosterone and estradiol were determined by Access immunoassay system and the morphological alterations in testis development observed by HE staining.

RESULTSThe body weight of the high-fat group obviously increased at the end of the 3rd week (P < 0.05), 26.6% heavier than that of the control by the end of the 6th week (P < 0.01), and Lee's index was also obviously increased (P < 0.01). Compared with the controls, the testicular coefficient declined in the high-fat group at the end of the 5th and 6th week (P < 0.05), plasma TG and TC remarkably increased, the testosterone level obviously decreased (P < 0.05), estradiol concentration lowered at the end of the 3rd, 4th and 5th week but dramatically increased at the 9th, with significant difference between the two groups (P < 0.01). Microscope examination showed that spermatogenic epithelial cells were arranged in disorder, the spermatogenic cell layers reduced and the number of mature sperms reduced.

CONCLUSIONHigh-fat diet can induce nutritional obesity in pubertal rats, which in turn may lead to the underdevelopment of the testis and the abnormal level of gonadal hormones.

Animals ; Body Fat Distribution ; Body Weight ; Diet Fads ; Epididymis ; growth & development ; pathology ; Male ; Obesity ; pathology ; physiopathology ; Organ Size ; Rats ; Rats, Sprague-Dawley ; Testis ; growth & development ; pathology

It has been demonstrated that insulin-like growth factor-1 (IGF-1) stimulates the proliferation and division of cells, facilitates the individual growth and development and regulates the material metabolism. Furthermore, it regulates male reproductive development and testicular endocrine functions. IGF-1 can stimulate the expression of GnRH gene in the hypothalamus of prepubertal male mice. However, it has no effect on or even inhibits GnRH gene expression in adult mice. IGF-1 may influence the growth, maturation and differentiation of GnRH neurons. It also accelerates LH and FSH secretion in hypophysis. IGF-1, produced locally in the testis and combined with its specific receptor, can regulate the proliferation and differentiation of adult Leydig cells, cause Sertoli cells to play different functions and control the biosynthesis of testicular hormones.
Animals ; Gene Expression Regulation ; Gonadotropin-Releasing Hormone ; biosynthesis ; Hypothalamo-Hypophyseal System ; metabolism ; physiology ; Insulin-Like Growth Factor I ; physiology ; Leydig Cells ; physiology ; Male ; Mice ; Sertoli Cells ; physiology ; Testis ; metabolism ; physiology

Objective To investigate the high-risk behaviors related to acquared immune deficiency syndrome/sexually transmitted disease (AIDS/STDs) infection among fishermen in Lv-si harbor,Jiangsu province.Methods A cross-sectional study was designed to investigate the research participants' demographic characteristics,high-risk behaviors that related to AIDS/STDs.Logistic regression was performed to measure the associations between potential risk factors and reported potential high-risk sexual behavior.Results 817 fishermen participated in the study and casual or commercial sex activities appeared to be the main high-risk behavior for AIDS/STDs infection in the target population.The rates of casual and commercial sex reported were 18.1％ and 28.9％ among fishermen.Risk factors associated with AIDS/STDs related high-risk behaviors among fishermen were high mobility (OR=1.516,P=0.038),higher lifetime sex frequency (OR=1.422,P=0.002)and unmarried status ( OR =7.527,P=0.014).Protective factors against high-risk behaviors were low intake of alcohol (OR=0.803,P=0.053),negative STD history (OR=0.268,P=0.001 ),age of initial sexual intercourse at or older than 22 years (OR =0.440,P=0.000) of age,as well as negative attitude toward multiple sexual parmers (OR=0.662,P=0.023 ) and legitimation for commercial sex (OR=0.612,P=0.007).Conclusion There were risk behaviors of AIDS/STDs in those infected fishmen.Casual and commercial sex were common high-risk behaviors.