Inhibition of Bcl-2 enhances the efficacy of epirubicin chemotherapy in PC-3 prostate cancer cells.

BACKGROUND: Overexpression of Bcl-2 protein in cancer cells can inhibit programmed cell death and engender chemoresistance. Bcl-2 antisense oligonucleotide (G3139) has shown its antitumor effects enhanced in preclinical models when combined with taxol-based chemotherapy. This study aimed to investigate the efficacy of G3139 combined with epirubicin in the androgen-independent prostate cancer. METHODS: PC3 prostate cancer cell line was cultured and treated with epirubicin and Bcl-2 antisense oligonucleotide alone or in combination. The effects of therapeutic agents on cells were determined by the MTT assay. Expression of Bcl-2 mRNA and protein was documented by RT-PCR and Western blotting. Apoptosis induction was confirmed by flow cytometric analysis. RESULTS: Bcl-2 antisense oligonucleotide alone produced no cytotoxic effects and the combination of Bcl-2 antisense oligonucleotide with epirubicin sensitized PC-3 cells to the killing effects of chemotherapy. A marked down-regulation of Bcl-2 mRNA and protein was observed after antisense and epirubicin cotreatment. A statistically significantly higher fraction of apoptotic cells was detected by flow-cytometric analysis after epirubicin treatment with prior antisense Bcl-2 transfenction, as compared with mono antisense Bcl-2 or epirubicin treatment. CONCLUSION: These data suggested that inhibition of Bcl-2 expression combined with epirubicin may be an attractive therapeutic strategy in hormone-refractory prostate cancer.

Downregulation of endothelin-1 by somatostatin improves liver function of recipients undergoing adult-to-adult living donor liver transplantation.

BACKGROUND: The aim of this study was to investigate the possible effect of somatostatin on the liver function of recipients undergoing living donor liver transplantation. METHODS: Forty recipients were randomized into group A (n = 20) and group B (n = 20). Recipients in group A received no somatostatin whereas somatostatin was administrated for recipients in group B perioperatively. Liver function, the plasma concentration of endothelin-1 and nitric oxide, the intragraft expressions of endothelin-1 and inducible nitric oxide syntheses at 2 hours after declamping of the portal vein were compared between the two groups. RESULTS: Compared to group A, alanine transaminase values in group B were significantly reduced at 2 hours after portal vein declamping, at the end of the operation and postoperation day 1 (P < 0.05), whereas aspartate aminotransferase values in group B decreased at 30 minutes after portal vein clamping, at 2 hours after portal vein declamping and at the end of the operation (P < 0.05). Total bilirubin values in group B were reduced significantly at 2 hours after portal vein declamping and at the end of the operation when compared to group A (P < 0.05). Intragraft expression of endothelin-1 was significantly downregulated at 2 hours after declamping of the portal vein accompanied with a reduction of plasma concentration of endothelin-1 in the peripheral blood (P < 0.05). CONCLUSIONS: Somatostatin had a protective effect on liver function during the early phase after declamping of portal vein for recipients undergoing living donor liver transplantation, and the possible mechanism might be partially attributed to the downregulation of endothelin-1.

Different plasma levels of interleukins and chemokines: comparison between children and adults with AIDS in China.

BACKGROUND: The immunological differences between children and adults with AIDS in China are not well documented. Th1/Th2 cytokines and chemokines are two types of immune factors intimately involved in disease progression of HIV-1 infection. This study aimed to identify changes in plasma levels of Th1/Th2 cytokines inerleukin (IL)-18, IL-16, IL-10 and chemokines regulated on activation, normal T cell expressed and secreted (RANTES), stromal cell-derived factor-1 (SDF-1) and monocyte chemoattractant protein-1 (MCP-1) in HIV-1-infected children and adults in China. METHODS: Seventy-five children with AIDS and 35 adult AIDS patients were recruited and clinical data were collected. CD4(+) T lymphocyte counts were measured by flow cytometery and plasma HIV RNA levels were measured by quantitative RT-PCR. Plasma levels of IL-18, IL-10, IL-16, RANTES, MCP-1, SDF-1alpha and SDF-1beta were quantified by enzyme-linked immunosorbent assay. The levels of beta2-microglobulin (beta2-MG) and soluble Fas (sFas) were measured to validate the level of humoral and cellular immune activation. RESULTS: The mean levels of all cytokines in pediatric and adult AIDS patients were significantly higher than in their healthy controls (P < 0.01). The mean levels of these cytokines were higher in pediatric patients than in adult patients (P < 0.05, except for SDF-1alpha and beta2-MG). Some of the cytokine levels in patients younger than 6 years old was higher than in older children and adults with AIDS (IL-10, IL-18, SDF-1alpha, MCP, RANTES and sFas, P < 0.05). Levels of IL-18, IL-10, RANTES and beta2-MG of pediatric patients increased as the levels of viral load increased (P < 0.05). CONCLUSIONS: Abnormal immune activation can be measured in Chinese pediatric and adult patients with AIDS, and is higher in children than in adult patients. The cytokines levels coincide with disease progression of AIDS, but have no direct relationship with total CD4(+) T cell count.

Calcitriol inhibits the growth of MHCC97 heptocellular cell lines by down-modulating c-met and ERK expressions.

BACKGROUND: Previous studies have demonstrated that Calcitriol or its analogs has an anti-tumour activity. This study was designed to determine the effect and mechanism of Calcitriol on MHCC-97 heptocellular cell lines. METHODS: MHCC97 cell lines were treated with Calcitriol of 10(-6) approximately 10(-9) M concentration and with Calcitriol in lipiodol ultra-fluid (LUF) respectively. The conditions of cell proliferation were analyzed by MTT method. The cell apoptosis and cycle were analyzed by using a flow cytometer. Hepatocyte growth factor (HGF) concentration in the cell supernatant was measured by using ELISA method. C-met and vitamin D receptor (VDR) mRNA in cells were determined by using RT-PCR method. VDR and ERK(1/2) proteins were determined by using Western Blotting method. RESULTS: Calcitriol inhibited the proliferation of MHCC97 cell lines with an accumulation of cells in the G(0)/G(1) phase and reduction of cells in S phase. Calcitriol dissolved in LUF resulted in a better and longer inhibitive effect on the cell lines than Calcitriol alone. MHCC97 cell lines secreted HGF and expressed c-met mRNA and ERK(1/2) proteins abundantly. Calcitriol remarkably inhibited the expressions of c-met mRNA and ERK(1/2) proteins. CONCLUSION: Calcitriol inhibits the growth of MHCC-97 heptocellular cell lines by down modulating c-met and ERK expressions.

[Impact of highly active antiretroviral therapy on plasma MCP-1 and MSP in AIDS patients].

OBJECTIVE: To study the effect of highly active antiretroviral therapy (HAART) on plasma levels of MSP and MCP-1 in AIDS patients. METHODS: Forty Chinese AIDS patients were treated with HAART for 3 months and 84 German AIDS patients with HAART for 3 to 6 years. The pre-treatment and post-treatment plasma levels of MSP and MCP-1 were measured by enzyme-linked immunosorbent assay (ELISA), and their correlations with CD4+ cell counts and viral loads were analyzed. RESULT: The mean levels of MCP-1 were significantly higher and MSP were significantly lower in HIV-infected patients compared with the HIV-negative controls (P <0.01). After HAART for three months, there were no significant changes in the levels of these cytokines. But after long-term HAART (for 3 to 6 y), the level of MCP-1 was increased and that of MSP decreased significantly (P<0.01). There was a negative correlation between MSP and MCP-1 levels, and the same for MSP level and CD4+ cell counts; while there was a positive correlation between MCP-1 levels and CD4+ cell counts. CONCLUSION: The changed plasma levels of MSP and MCP-1 are associated with HIV-1 infection and HAART may reverse the levels of these two cytokines.

Study of Resistin gene expression in peripheral blood mononuclear cell and its gene polymorphism in a small range population.

OBJECTIVE: To observe the expression of Resistin mRNA in peripheral blood mononuclear cells and its gene polymorphism in coding region in a small range population in Zhejiang Province of China. METHODS: Eighty-three cases of type 2 diabetes mellitus and 53 healthy people were included. The expression of Resistin mRNA in peripheral blood mononuclear cells was detected by RT-PCR and semi-quantitative PCR assay. The sequencing work was done in Resistin cDNA and gene polymorphism was analyzed. RESULTS: At the same condition, in 83 diabetes patients, Resistin mRNA was detected in 23 cases (11 males and 12 females). There was no Resistin mRNA expression in 53 healthy people. The ratio of PCR products between Resistin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was from 0.564 to 1.238, averaging 0.804+/-0.436. The sequence of Resistin cDNA is almost identical with each other and with that in GenBank with no single nucleotide polymorphism being found. CONCLUSION: Resistin mRNA is expressed in human peripheral blood mononuclear cells in some type 2 diabetes mellitus, but its expression is at a low level. Among the experiment population we did not find polymorphism phenomenon in Resistin coding region. The different individual's Resistin coding region is highly coincident.

[Comparison of immunological profiles between pediatric and adult patients with AIDS in China].

OBJECTIVE: To compare the immunological profiles of pediatric and adult patients with AIDS in China. METHODS: Totally 103 pediatric AIDS patients, 38 adult patients, 88 healthy children, and 72 healthy adults were enrolled. CD4 + T lymphocyte counts were determined by four-color flow cytometer and HIV-RNA levels were measured in EDTA plasma by quantitative reverse-transcription polymerase chain reaction (RT-PCR). Plasma levels of interleukin (IL)-10, IL-16, IL-18, regulated upon activation, normal T-cell expressed and secreted (RANTES), monocyte chemoattractant protein 1 (MCP-1), stromal cell-derived factor-(SDF-1) alpha, SDF-1 beta, and macrophage stimulate protein (MSP) were quantified by enzyme-linked immunosorbent assay (ELISA). The levels of beta 2-microglobulin (beta 2-MG) and soluble Fas (sFas) were measured to indicate the activation of immune system. RESULTS: The mean CD4 + T cell count in pediatric patients with AIDS was significantly lower than in healthy children (P < 0.01), as between the adult AIDS patients and healthy adults (P < 0.01). The mean levels of these cytokines in pediatric patients were significantly higher than in healthy children (P < 0.01). The level of MSP in adult patients was significantly lower than in healthy adults and other cytokines were significantly higher (P < 0.01). The mean levels of these cytokines, except SDF1 alpha and beta 2-MG, were significantly higher in pediatric patients than in adult patients (P < 0.01). CONCLUSION: Abnormal immune activation is induced in both pediatric and adult patients with HIV-1 infection. The level of immune activation is higher in pediatric patients than in adult patients.

[Study on the prognostic value of hepatocyte growth factor and c-met for patients with hepatocellular carcinoma].

OBJECTIVE: To analyze the prognostic value of hepatocyte growth factor (HGF) and c-met for patients with hepatocellular carcinoma (HCC) after hepatectomy. METHODS: Twenty-five patients undergoing partial hepatectomy for HCC were studied. Serum HGF level was determined using ELISA kit before and after operation respectively. c-met protein and mRNA expression in cancerous and paracancerous tissues were detected by immunohistochemical and RT-PCR methods respectively. The correlations of clinical-pathologic parameters with the HGF level in serum and c-met expression in cancerous tissue were analyzed respectively. RESULTS: HCC patients had a significantly higher concentration of serum HGF than normal controls and chronic hepatitis B respectively [(1.03 +/- 0.09) ng/ml vs (0.69 +/- 0.02) ng/ml and (0.74 +/- 0.09) ng/ml]. No significant difference in serum HGF was observed between HCC and cirrhosis patients with Child-Pugh score B/C [(1.03 +/- 0.09) ng/ml vs (1.04 +/- 0.11) ng/ml]. Serum HGF concentrations were positively correlated with tumor size (> 5 cm), node cirrhosis, portal vein tumor thrombi (PVTT) and preoperative alpha-fetoprotein (AFP) level (> or = 400 microg/L). After the resection of tumor, serum HGF concentration had a peak on the third postoperative day (POD), and then declined, but did not return to normal level on the tenth POD. From preoperative day to third POD, HGF concentration had a higher elevation in patients with major resection than with local resection. Moderately or strongly positive expression of c-met protein was observed in 21 cancerous regions (21/25), and only in 5 paracancerous regions. The intensive expression of c-met mRNA was 100% (25/25) detectable in the cancerous tissues, but only 24% (6/25) in the paracancerous tissues. The expression extent of c-met protein was correlated with portal vein tumor thrombi (PVTT). In paracancerous tissues, the expression of c-met protein was more intense in patients with cirrhosis than those without cirrhosis. The patients with recurrence or metastases after operation had a higher level of serum HGF and more intensive expression of c-met than other patients. No significant association was observed between HGF in serum and c-met expression in cancerous tissue. CONCLUSIONS: The over-expression of HGF and its receptor c-met indicate an adverse prognosis for HCC patients. The sustained high level of serum HGF after hepatectomy may be a factor related to early tumor recurrence and metastasis.

[Atherosclerosis and androgen levels in elderly males].

OBJECTIVE: To observe the relation of androgen levels and atherosclerosis (AS) in elderly males. METHODS: Both carotid arteries and arteries of lower extremity were examined with Doppler ultrasonography. Those with arteriosclerosis and much atheromatous plaque were designated as case group, and those with normal results formed control group. Total testosterone (TT), free testosterone (FT) and estradiol (E2) were measure by radioimmunoassay, HDL-C, LDL-C, TC and TG were assayed by colorimetry, vascular endothelium growth factor (VEGF), tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), intercellular adhesion molecule (sICAM-1) were determined by ELISA. RESULT: FT was significantly lower in case group than in control group (P<0.01), no differences were found in TT, E2. HDL-C in control group was higher than that in case group (P<0.01), TC and TG were higher in case group than those in control group (P<0.05). HDL-C was correlated positively and LDL-C was negatively with FT level, while both TC and TG in case group had negative relation with FT. VEGF was higher in case group (P<0.05), and it had negative relation with FT in both groups. TNF-alpha and IL-6 were significantly higher in case group (P<0.05), and they had negative relation with FT. sICAM-1 was significantly lower in control group than it in case group (P<0.01). CONCLUSION: The normal androgen levels, especially FT, have beneficial effect in AS development in elderly males. Low FT level may be an independent risk factor in AS development.

Study of androgen and atherosclerosis in old-age male.

OBJECTIVE: To observe the difference of androgen and inflammatory cytokines level in atherosclerosis and analyse their relations. METHOD: Both carotid arteries and arteries of lower extremity were subjected to ultrasonic examination by Doppler's method. Those with much atheromatous plaque formation were ranged into case group, and those with normal result formed control group. Total, free testosterone and estradiol were assayed by radioimmunoassay. C reactive protein (CRP) was assayed by nepheloturbidity. Tumor necrosis factor-alpha (TNF-alpha), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-18 (IL-18), soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1) were assayed by ELISA. The mean difference between two groups and the correlation between free testosterone and cytokines were analysed. RESULTS: Free testosterone was (6.337+/-3.371) pg/L in case group and (11.375+/-4.733) pg/L in control group, P<0.01. No differences were found in total testosterone and estradiol. CRP was (27.294+/-10.238) mg/L in case group and (12.843+/-6.318) mg/L in control group, P<0.01. IL-6 was (41.700+/-31.385) pg/L in case group and (25.396+/-20.772) pg/L in control group, P<0.05. IL-8 was (89.249+/-58.357) pg/L in case group and (67.873+/-31.227) pg/L in control group, P<0.05. sICAM-1 was (470.491+/-134.078) pg/L in case group and (368.487+/-97.183) pg/L in control group, P<0.01. sVCAM-1 was (537.808+/-213.172) pg/L in case group and (457.275+/-157.273) pg/L in control group, P<0.05. There were no differences in TNF-alpha, IL-10 and IL-18. Correlation analysis showed that FT (free testosterone) had negative correlation with CRP, IL-6 and sICAM-1. Among them FT had well correlation with CRP, correlation index was -0.678. CONCLUSION: Free testosterone was in negative correlation with atherosclerosis in old-age male. Free testosterone may have the role of anti-atherosclerosis, and this effect was not achieved by its transformation to estradiol. Low free testosterone level was followed by increased level of inflammatory cytokines. Low free testosterones coexist with inflammation and they both affect the process of atherosclerosis in old-age male.