[Show abstract][Hide abstract] ABSTRACT: Leukemia related protein 16 (LRP16) was first cloned from acute myeloid leukemia cells in our laboratory. In the present study, we sought to investigate the role of LRP16 in insulin action and sensitivity, using LRP16-depleted and -overexpressing 3T3-L1 cells. LRP16 silencing resulted in a reduction of the expression and secretion of tumor necrosis factor-alpha (TNF-α) and a concomitant increase in the expression of peroxisome proliferator-activated receptor-gamma (PPAR-γ). Moreover, LRP16 depletion promoted insulin-induced glucose uptake and adipocyte differentiation of 3T3-L1 cells. In contrast, LRP16 overexpression increased TNF-α secretion, suppressed glucose uptake, and attenuated 3T3-L1 cell differentiation. The phosphorylation levels of insulin receptor substrate 1 (IRS-1), phosphatidylinositide 3-kinase (PI3-K), and Akt were increased in LRP16-deficient 3T3-L1 cells, and conversely, diminished in LRP16-overexpressing 3T3-L1 cells, when compared to the corresponding control cells. Additionally, LRP16 overexpression raised the phosphorylation level of mammalian target of rapamycin (mTOR). The pretreatment with rapamycin, a specific inhibitor of mTOR, prevented the TNF-α elevation and PPAR-γ reduction and restored the phosphorylation of IRS-1, PI3-K, and Akt in LRP16-overexpressing cells. Our data collectively indicate that LRP16 acts as a negative regulator of insulin action and adipogenesis in 3T3-L1 adipocytes, which involves the activation of the mTOR signaling pathway.
Hormone and Metabolic Research 02/2013; · 2.15 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The balance between interleukin-18 (IL-18) and its endogenous antagonist, IL-18 binding protein (IL-18BP), was evaluated in children with Henoch-Schönlein purpura (HSP). Plasma IL-18 and IL-18BP levels and peripheral blood mononuclear cell IL-18 mRNA expression were significantly higher in patients with active HSP (n = 30) than in healthy controls (n = 20); IL-18BP mRNA expression was similar in active HSP and controls. Plasma levels and mRNA expression of IL-18 and IL-18BP in patients in remission (n = 19) were similar to those in controls. The ratios of IL-18 / IL-18BP plasma levels and IL-18 / IL-18BP mRNA levels in active HSP were significantly higher than in patients in remission and healthy controls. Thus, adequate IL-18BP to block the proinflammatory activity of IL-18 may not be present in active HSP and regulation of the IL-18 / IL-18BP balance might provide a potential therapeutic strategy.
The Journal of international medical research 12/2011; 39(6):2201-8. · 1.10 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Esophageal cancer (EC) is a common malignant cancer threatening people's health. There are no universally accepted parameters for its early diagnosis. The aim of this study was to observe the expression of human leukocyte antigen class I (HLA-I) on peripheral blood mononuclear cells (PBMCs) of EC patients and in individuals of high-incidence area of EC so as to evaluate the feasibility of using this parameter as a potential non-invasive biomarker for the early diagnosis of EC. The present study enrolled 58 pathological confirmed EC patients, 46 patients with benign esophageal disease, and 65 healthy volunteers. Expression levels of HLA-I protein and mRNA on PBMCs were determined by flow cytometry and quantitative reverse transcriptase polymerase chain reaction, respectively. Then, 181 volunteers from Lijiadian, a village with high morbidity of EC, and 153 age- and gender-matched health volunteers were involved in this study to observe HLA-I expressions in individuals of high-incidence area of EC. Compared with benign esophageal disease and health volunteers, the expressions of HLA-I protein and mRNA on PBMCs of EC patients are significantly decreased, especially in patients with stage III and IV EC, but was not influenced by patient's age and gender. Furthermore, individuals of high-incidence area of EC also show downregulated HLA-I protein, but not mRNA, expression on PBMCs. Altogether, HLA-I expression on PBMCs of EC patients and individuals from high-incidence area of EC is downregulated, and this parameter might be used as a potential predictor of EC.
Diseases of the Esophagus 09/2011; 25(3):273-8. · 2.06 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This study investigated major histocompatibility complex class I (MHC-I) antigen expression on peripheral blood T-cells after transplantation to assess its potential as an early marker of acute graft rejection (AGR). Using a mouse model with or without immunosuppressive treatment, the expression of MHC-I antigens on CD3(+)CD8(+) T-lymphocytes was assessed by flow cytometry following syngeneic graft (n = 138) or allograft (n = 138) skin transplantation. The occurrence of AGR was assessed by examining the degree of lymphocyte and monocyte infiltration in transplant biopsies. During AGR, expression of MHC-I antigens increased significantly compared with pre-transplant levels in the allograft group, even with immunosuppressive treatment. The highest expression of MHC-I antigens occurred 5 - 6 days before macroscopic rejection. These results suggest that expression of MHC-I antigens on peripheral blood CD3(+)CD8(+) T-lymphocytes could be used as an early marker for predicting AGR.
The Journal of international medical research 04/2011; 39(2):480-7. · 1.10 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Human leucocyte antigen class I (HLA-I), which includes HLA-A, -B and -C, is an essential immune factor participating in the antitumour immune response. The changes in HLA-I expression in peripheral blood T lymphocytes in cancer patients have yet to be defined. This study examined the expression of HLA-I on CD4(+) and CD8(+) T lymphocytes in female patients with stage I - IV breast infiltrating ductal carcinoma, benign breast tumour diseases (mammary intraductal papilloma or breast fibroadenoma), and in healthy controls. HLA-I was down-regulated on CD4(+) T lymphocytes from patients with stage III and IV cancer, and on CD8(+) T lymphocytes in patients with stage I - IV cancer compared with healthy controls. HLA-I expression in T lymphocytes may contribute towards immune-balance disorders in tumour patients.
The Journal of international medical research 04/2011; 39(2):508-13. · 1.10 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To investigate the correlation between abnormal oral glucose tolerance test (OGTT) 30-min plasma glucose (PG) and risk factors and metabolic abnormalities of diabetes and cardiovascular diseases (CVD).
Participants (no.=2457) underwent a physical examination, blood biochemistry examination, OGTT, and a 12-lead electrocardiogram. We measured fasting PG (FPG) and PG at 30 min (30minPG) and 1 h (1hPG) of the OGTT. The association between an increase in 30minPG (by 1 SD) and the metabolic abnormalities of diabetes and CVD such as hypertension, overweight and obesity, central obesity, and hyperlipidemia; osteoporosis was assessed by logistic regression analysis after controlling for FPG and 2hPG.
This analysis showed that an increase of 30minPG by 1 SD (1.92 mmol/l) significantly increased the risk of chronic metabolic abnormalities in diabetes and CVD such as hypertension, overweight, and obesity, central obesity, hyperlipidemia, and osteoporosis. Stepwise multiple regression analysis also showed that 30minPG was significantly correlated with male gender, smoking, FPG, 2hPG, total cholesterol, waist/hip ratio, and blood pressure.
An elevation of 30minPG increased the risk of diabetes and CVD. The increased risk was independent of FPG, 2hPG, age, sex, and smoking status.
Journal of endocrinological investigation 10/2010; 34(5):e115-20. · 1.65 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This study was designed to investigate whether there is a correlation between the down-regulation of microRNA-218 (miR-218) and the presence of human papillomavirus (HPV) infection in the pathogenesis of cervical cancer. The participants comprised 78 women with cervical intraepithelial neoplasia (CIN); 22 (28.2%) had CIN 1, 27 (34.6%) had CIN 2 and 29 (37.2%) had CIN 3. MiR-218 expression was determined by reverse transcriptase polymerase chain reaction and HPV genotypes in tissue specimens were identified with a microarray test kit. The findings showed that miR-218 levels in patients with high-risk HPV infection were lower than in those infected with low-risk or intermediate-risk HPV, or in those who were HPV-free. MiR-218 levels in patients with high-risk CIN were lower than in those with low-risk CIN. We concluded that infection with high-risk HPV lowered the expression of miR-218 and that down-regulation of miR-218 was involved in the pathogenesis of cervical cancer.
The Journal of international medical research 10/2010; 38(5):1730-6. · 1.10 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This study compared the expression of the genes encoding human leucocyte antigens (HLA)-A, -B, -DP, -DR and -G in peripheral blood mononuclear cells (PBMCs) in gastric cancer patients and healthy controls. Using reverse transcription-polymerase chain reaction, levels of classical HLA-A, -B, -DP and -DR and non-classical HLA-G mRNA were studied in 43 gastric cancer patients and 22 controls. In addition, the levels of HLA-A,B,C and -G antigens on the surface of PBMCs were measured in 30 gastric cancer patients and 15 controls using flow cytometry. The mean fluorescence intensity of HLA-A,B,C antigen in the gastric cancer group was significantly lower than in controls. The HLA-G antigen was mainly present on CD4(+)CD8(-) T-lymphocytes. The percentage of CD4(+)CD8(-) T-lymphocytes positive for HLA-G antigen was significantly lower in the gastric cancer group compared with the healthy controls. Levels of HLA-A, -B and -G mRNA in the gastric cancer group were significantly lower than in controls. The HLA-G mRNA levels were significantly lower in gastric cancer of histological grades III and IV than in grades I and II. These data may provide a novel diagnostic and research tool for gastric cancer.
The Journal of international medical research 05/2010; 38(3):949-56. · 1.10 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The goal of this study was to determine the prevalence of diabetes mellitus (DM), impaired glucose regulation (IGR) and related metabolic disorders (overweight, obesity and hypertension) in a Chinese population (20-74 years old). An additional goal was to investigate the relationship between glucose metabolism and the Minnesota code-indicated major abnormal electrocardiogram (MA-ECG). There were 3960 individuals selected from urban and rural areas of Fujian, China from July 2007 to May 2008 by multistage-stratified sampling. Ultimately, data from 3208 subjects (20-74 years old) were analysed (including physical measurements, blood biochemical analysis, oral glucose tolerance test and 12-lead resting ECG). According to World Health Organization diagnostic criteria, the prevalence rates of DM and IGR were 9.51% (male, 10.08%; female, 9.14%) and 14.40% (male, 14.48%; female, 14.35%) respectively. Newly diagnosed DM was found in 53.44% of the diabetic subjects. Based on the 2000 China census, the age-standardized prevalence rates of DM and IGR were 7.19% (male, 7.74%; female, 6.61%) and 11.96 % (male, 12.35%; female, 11.56%) respectively. The age-standardized prevalence rates of DM and IGR in urban areas (7.74% and 12.97% respectively) were slightly but no significantly higher than in rural areas (6.67%, 10.86%). The prevalence rates of overweight, obesity and hypertension were 25.50%, 3.52% and 28.52% respectively (age- and sex- standardized rates: 23.69%, 3.02 % and 22.45 %). After adjusting for other confounding risk factors, multiple logistic regression analysis showed that DM and impaired glucose tolerance were independent risk factors for MA-ECG. Non-diabetic subjects with increased 30-min plasma glucose (PG) after an oral glucose load had a higher risk of MA-ECG after adjusting for other risk factors, especially in those with normal glucose tolerance but with 30-min PG >or= 7.8 mmol L(-1) (odds ratio = 1.371 [1.055-1.780]). The prevalence rates of DM and IGR as well as other metabolic disorders have increased dramatically in the last decade in China, especially in rural areas, with many undiagnosed cases of DM. Even slightly elevated PG levels may predict early cardiovascular events.
[Show abstract][Hide abstract] ABSTRACT: To investigate the clinical and pathological characteristics of thyroid nodules, as well as to evaluate the significance of ultrasonographically detected thyroid calcification in the diagnosis of thyroid carcinomas.
Retrospective data were studied from 1,051 consecutive patients who underwent a thyroidectomy in the Provincial Hospital of Fujian Medical University in South China between January 2003 and July 2006 for nodular thyroid disease. Complete sonographical information before surgery was only collected from 758 of the 1,051 patients.
Among the 1,051 patients, benign lesions were found in 857 (81.54%) patients, of whom 612 (71.41%) were nodular goiter; malignant lesions were found in 194 (18.46%) patients, in whom benign thyroid lesions were also found in 85 (43.81%) patients. A total of 48 patients suffered from microcarcinomas, of whom 37 patients had benign lesions; these 37 accounted for 43.53 and 77.08%, respectively, of the 85 malignant cases with benign lesions and the 48 cases with microcarcinomas. In the 758 patients who underwent thyroid ultrasonography before surgery, intrathyroidal calcifications were apparent in 243 patients (32.06%). The incidence of calcification was significantly higher in patients with thyroid carcinoma (54.17%) than in those with benign lesions (26.87%; p < 0.005). Detection of calcification in thyroid lesions by ultrasound had a sensitivity of 32.38% and a specificity of 87.35%, with an OR of 3.31 (95% CI, 2.24-4.63), positive likelihood ratio of 2.56, negative likelihood ratio of 0.77 and a kappa value of 0.23.
Thyroid carcinoma, especially microcarcinoma, often coexists with benign thyroid disease. Calcification detected by thyroid ultrasound represents a risk factor for malignancy, but is of limited use as a sole marker of malignancy.
European Surgical Research 01/2009; 42(3):137-42. · 1.43 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Diabetes is a major independent risk factor for cardiovascular disease and stroke. High glucose (HG) reduces endothelial cell (EC) proliferation with a concomitant increase in apoptosis. HG also induces the translocation of nuclear factor (NF)-kappaB in human umbilical vein endothelial cells (HUVECs). However, data regarding the relationship between NF-kappaB signaling and HG-induced endothelial dysfunction are limited. In the present study, we constructed an NF-kappaB-targeting RNA interference (RNAi) adenovirus vector and cultured HUVECs in 5.5, 20.5, or 30.5 mM D: -glucose or in daily alternating 5.5 or 30.5 mM D: -glucose. We assessed the effects of the NF-kappaB pathway on proliferation under HG conditions by measuring bromodeoxyuridine incorporation and conducting methyl thiazolyltetrazolium assays. We also tested apoptosis by performing flow cytometry and terminal deoxynucleotidyl transferase nick-end labeling assay. The RNAi adenovirus effectively downregulated expression of the p65 protein in HUVECs for more than 6 days. Blockage of the NF-kappaB pathway with the RNAi adenovirus substantially protected HUVECs from decreased proliferation and reduced cellular apoptosis in HG conditions. These findings may explain how hyperglycemia promotes dysfunction of ECs and could elucidate a potential new target for therapeutic interventions.
[Show abstract][Hide abstract] ABSTRACT: To construct RNAi recombinant adenoviral expressive vectors specific to glycogen synthase kinase-3beta (GSK-3beta) and to observe its gene knockdown effect on the expression of GSK-3beta, and to explore the effect of Wnt/beta-catenin pathway on the proliferation of human thyrocytes using the RNAi adenovirus vector.
An adenovirus plasmid that contained the RNAi cassette targeting the GSK-3beta gene was constructed by homologous recombination and cloning techniques, transfected into human embryo kidney (HEK) 293A cells to product adenovirus, and then was used to infect the HEK293A cells to amplify the adenoviral stock. Plaque forming assay was used to titer the adenoviral stock. Normal human thyrocytes fart from thyroid adenoma were obtained during resection of adenoma, cultured, and infected by the GSK-3beta specific RNAi adenovirus. The GSK-3beta gene silencing effect induced by the RNAi adenovirus was detected by Western blotting 0, 24, 48, 72, 120, and 144 hours later. BrdU method was used to detect the cell proliferation. Another HEK293A cells were divided into 3 groups: infected with recombinant adenovirus plasmid Ad-1457, infected with un-recombinant framework plasmid pAd-DEST, and un-infected. 72 hours later Western blotting was used to examine the level of beta-catenin.
The GSK-3beta expression of the thyrocytes infected with the recombinant adenovirus plasmid Ad-1457 were significantly lower than those of the thyrocytes infected with Ad-DEST (all P<0.05). The expression of beta-catenin of the thyrocytes infected with Ad-DEST was significantly higher than those of the Ad-DEST group and un-infected group (both P<0.05). BrdU assay suggested that the proliferation rates 1, 3, 5, and 7 days after infection of the thyrocytes infected with Ad1457 plasmid were significantly higher than those of the thyrocytes infected with the plasmid pAd-DEST (all P<0.05).
RNAi adenovirus is an important tool inhibiting the expression of target gene efficiently. The Wnt/beta-catenin pathway plays an important role in the regulation of proliferation of human thyrocytes.
[Show abstract][Hide abstract] ABSTRACT: Background and aimNotch signaling controls cellular differentiation and proliferation. Deregulated expression of Notch receptors is observed in a growing number of malignant tumours, however, the role of Notch signaling in hepatocellular carcinoma is still unknown. To address this, the expression of Notch receptors in human hepatocellular carcinoma was examined in both protein and ribonucleic acid levels.Patients and methodsFifty-three hepatocellular carcinoma tissue sections were detected by immunohistochemistry. Three paired fresh surgical hepatocellular carcinoma and adjacent nontumour liver samples were analyzed by Western blot and reverse transcriptase polymerase chain reaction. Immunohistochemistry, Western blot and reverse transcriptase polymerase chain reaction are reliable methods to examine the expression of protein and RNA.ResultsAll of the four Notch receptors were expressed in the neoplastic cells of hepatocellular carcinoma tissues with different intensity and extensity. Notch1 and Notch4 were expressed in both cytoplasm and nucleus, and all of the nuclear staining showed up in the cytoplasm-positive cases. Cytoplasmic and nuclear Notch1 was detected in 88.7% (47/53) and 9.4% (5/53) of hepatocellular carcinoma tissues, respectively; positive rates of Notch4 were 67.9% (36/53) in cytoplasm and 52.8% (31/53) in nucleus. Notch2 and Notch3 were only in cytoplasm, with positive rates of 26.4% (14/53) and 52.8% (28/53), respectively. Compared with adjacent nontumour liver, Notch1 (cytoplasmic) and Notch4 (nuclear) were up-regulated (P < 0.05, P < 0.05), Notch2 was down-regulated (P < 0.05), while Notch1 (nuclear), Notch3 and Notch4 (cytoplasmic) showed no difference between hepatocellular carcinoma and adjacent nontumour liver. Western blot and reverse transcriptase polymerase chain reaction analysis showed a consistent result.ConclusionOur findings indicate that the expression of Notch receptors was deregulated and Notch signaling might be involved in the development of hepatocellular carcinoma.
Digestive and Liver Disease 02/2008; · 2.89 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Early diagnosis of acute graft rejection is important in the clinic. To explore a reliable diagnostic marker, we selected skin-grafted rabbits as an animal model to study peripheral blood mononuclear cell (PBMC) major histocompatibility complex 1 (MHC I) and MHC II gene mRNA in acute graft rejection (AGR).
Fifteen New Zealand white rabbits were randomly divided into three groups to observe skin graft rejection: three rabbits were in the autograft control group; six rabbits in a cyclosporine (CsA) treated allografted group; and the other six rabbits in untreated allografted group. The CsA-treated allografted group was given CsA (5 mg/kg) daily intramuscularly. PBMC samples were obtained every 2 days to detect by real-time polymerase chain reaction, PBMC MHC I and MHC II gene mRNA.
MHC I and MHC II gene mRNA levels did not show any obvious change in the autografted controls. MHC I gene mRNA levels showed a slow increase in the CsA-treated allografted group, but no obvious change in the untreated allografted group. MHC II gene mRNA reached the highest level at 2 to 3 days before graft rejection appeared macroscopically in the CsA-treated allografted group and untreated allografted group, then decreasing to a low level.
Compared with MHC I gene mRNA expression, PBMC MHC II gene mRNA expression may be considered to be an earlier marker for AGR.
[Show abstract][Hide abstract] ABSTRACT: The overexpression of a new cytokine-induced apoptosis inhibitor 1 (CIAPIN1) gene has been shown previously to promote a multidrug
resistant phenotype in gastric cancer cells through the upregulation of MDR1 and MRP1. In the present study, we constructed
the siRNA eukaryotic expression vectors of CIAPIN1 and transfected them into SGC7901/VCR cells to examine whether the downregulation of CIAPIN1 increased cell sensitivity towards chemotherapeutic drugs. After transfection, the expression of CIAPIN1 was dramatically decreased in CIAPIN1 siRNA transfectants compared with that in parental cells and empty vector control cells. The downregulation of CIAPIN1 significantly enhanced the sensitivity of SGC7901/VCR cells to vincristine (VCR), adriamycin (ADR), and etoposide (VP-16),
but not to 5-fluorouracil (5-Fu) and cisplatin (CDDP). Cell capacity to efflux adriamycin decreased markedly in CIAPIN1 siRNA transfectants, and the correlation between CIAPIN1 downregulation and decreased MDR-1 transcriptional activity was
observed. CIAPIN1 siRNA could significantly downregulate the expression of Bcl-2, and upregulate the expression of Bax, but does not alter
the expression of PTEN in gastric cancer cells. These observations suggested that the siRNA constructs of CIAPIN1 we obtained could effectively downregulate the expression of CIAPIN1 and reverse the resistant phenotype of gastric cancer cells. Further study of the biological functions of CIAPIN1 may be
helpful for understanding the mechanisms of multidrug resistance of gastric cancer and in developing possible strategies to
treat gastric cancer.
–multidrug resistance–apoptosis–gastric cancer–RNAi
[Show abstract][Hide abstract] ABSTRACT: The overexpression of a new cytokine-induced apoptosis inhibitor 1 (CIAPIN1) gene has been shown previously to promote a multidrug resistant phenotype in gastric cancer cells through the upregulation of MDR1 and MRP1. In the present study, we constructed the siRNA eukaryotic expression vectors of CIAPIN1 and transfected them into SGC7901/VCR cells to examine whether the down regulation of CIAPIN1 increased cell sensitivity towards chemotherapeutic drugs. After transfection, the expression of CIAPIN1 was dramatically decreased in CIAPIN1 siRNA transfectants compared with that in parental cells and empty vector control cells. The down-regulation of CIAPIN1 significantly enhanced the sensitivity of SGC7901/VCR cells to vincristine (VCR), adriamycin (ADR) and etoposide (VP-16), but not to 5-fluorouracil (5-FU) and cisplatin (CDDP). Cell capacity to efflux adriamycin decreased markedly in CIAPIN1 siRNA transfectants, and correlation between CIAPIN1 down regulation and decreased MDR1 transcriptional activity were observed. CIAPIN1 siRNA could significantly down regulate the expression of Bcl-2, and up-regulate the expression of Bax, but not alter the expression of PTEN in gastric cancer cells. These observations suggested that the siRNA constructs of CIAPIN1 we obtained could effectively down-regulate the expression of CIAPIN1 and reverse the resistant phenotype of gastric cancer cells. The further study of the biological functions of CIAPIN1 may be helpful for understanding the mechanisms of multidrug resistance of gastric cancer and developing possible strategies to treat gastric cancer.