Overexpression of glucosylceramide synthase and its significance in the clinical outcome of non-small cell lung cancer

Background Glucosylceramide synthase （GCS）,an enzyme responsible for ceramide glycosylation,plays an important role in multidrug resistance （MDR） in some tumors in vitro; however,its expression and clinicopathological significance in non-small cell lung cancer （NSCLC） remains unclear.Methods We evaluated GCS expression in 116 paired tumor and adjacent non-cancerous tissues and 50 frozen tissues from patients with NSCLC using immunohistochemistry and western blotting,and explored the correlation between GCS and NSCLC clinicopathological characteristics and prognosis.We observed the association between GCS and the MDR proteins P-glycoprotein （P-gp） and lung resistance-related protein （LRP） to determine the link between GCS and MDR at the histological level.Results GCS expression was significantly upregulated in NSCLC tumors compared with non-cancerous tissue.There was high GCS expression in 75/116 tumor specimens （64.7％） and 16/116 non-cancerous specimens （13.8％）.High GCS expression was significantly associated with poor differentiation （P=0.01）,lymph node metastasis （P=0.004）,recurrence/ distant metastasis （P=0.006）,and chemotherapy resistance （P=0.025）.Multivariate analysis demonstrated that GCS immunopositivity was an independent risk factor for survival （P=0.018）.P-gp was expressed in 80/116 tumors （69.0％） and in 12/116 non-cancerous tissue specimens （10.3％; P=0.001）; LRP was expressed in 85/116 tumors （73.3％） and 19/116 non-cancerous tissue specimens （16.4％; P=0.001）.Importantly,the results demonstrated that increased GCS expression in NSCLC cancer specimens correlated with increased expression of P-gp and LRP,molecules known to stimulate cancer cell MDR （r=0.612 and 0.503,P=0.01 and 0.035,respectively）.Conclusion GCS upregulation might contribute to the development of NSCLC and could be a useful prognostic indicator and chemoresistance predictor for NSCLC patients.     

Expressions of farnesoid X receptor and myeloid cell leukemia sequence 1 protein are associated with poor prognosis in patients with gallbladder cancer

Background Farnesoid X receptor （FXR） regulates tumorigenesis, but its clinical significance in gallbladder cancer （GBC） remains unclear. This study investigated its clinical and prognostic significance in GBC patients, as well as its association with the anti-apoptotic protein, myeloid cell leukemia sequence 1 （MCL1） protein. Methods FXR and MCL1 expression in 42 primary GBC and 15 normal gallbladder tissues were analyzed by immunohistochemistry. The patients and samples were collected from Ren Ji Hospital from January 2005 to December 2010. Their association with clinicopathologic factors and prognosis, as well as the correlation between FXR and MCL1 protein expression were analyzed by statistical analyses. Results Compared with normal gallbladder tissues, FXR expression was decreased and MCL1 expression was increased in GBC, during progression of tumor node metastasis （TNM） stage. The Kaplan-Meier survival analysis showed that FXR low-expression and MCL1 over-expression were significantly associated with overall poor survival. Furthermore, multivariate analysis showed that FXR and MCL1 are both prognostic factors for GBC patients. FXR low-expression was significantly correlated with MCL1 over-expression. Conclusion FXR might be a new molecular marker to predict the prognosis of patients with GBC and a novel therapeutic target. Chin Med J 2014;127 （14）： 2637-2642     

Angiogenesis effect of Astragalus polysaccharide combined with endothelial progenitor cells therapy in diabetic male rat following experimental hind limb ischemia

Background Diabetes mellitus （DM） is a common disease accompanied with a high incidence of hind limb ischemia （HLI）.In recent years,numerous studies demonstrated that endothelial progenitor cells （EPCs） are involved in angiogenesis and maintenance of vascular integrity following HLI.On the other side,it has been proved that Astragalus polysaccharide （APS） could promote angiogenesis.In the present study,we aimed to evaluate the effect of APS and EPCs on enhancing angiogenesis after experimental HLI caused by femoral artery ligation in rats with streptozotocin （STZ）-induced diabetes.Methods Rats （n=110） were randomly assigned to the following groups：sham group,ischemia group,APS group,EPCs group and APS＋EPCs group.APS,EPCs or an equal volume of vehicle was administered intramuscularly after HLI induction,and 6 rats were assessed by angiography at 28 days after induction of HLI,6 rats were sacrificed at the same time point to take histological studies,biochemical tests were also performed at that point in the rest rats.Results APS or EPCs treatment induced an increase,respectively,in the protein expression of vascular endothelial growth factor （VEGF） （36.61%,61.59%）,VEGF receptor-1 （VEGFR-1） （35.50%,57.33%）,VEGFR-2 （31.75%,41.89%）,Angiopoietin-1 （Ang-1） （37.57%,64.66%） and Tie-2 （42.55%,76.94%） （P 〈0.05）,after HLI injury.And combined therapy of APS and EPCs enhanced the effort of angiogenesis after HLI induction in diabetic rats,through elevating protein expression of VEGF （99.67%）,VEGFR-1 （105.33%）,VEGFR2 （72.05%）,Ang-1 （114.30%） and Tie-2 （111.87%） （P〈0.05）.Similarly,mRNA expression of VEGF,VEGFR-1,VEGFR2,Ang-1,Tie-2 also show similar trends as well as protein expression （P〈0.05）.Conclusion APS or EPCs could enhance angiogenesis,and the combined treatment leads to better effort,at least,partially via VEGFNEGFR and Ang-1/Tie-2 signaling pathway.     

miR-30b and miR-30c expression predicted response to tyrosine kinase inhibitors as first line treatment in non-small cell lung cancer

Background Aberrantly expressed microRNAs are a hallmark of cancer,and microRNA expression profiling is associated with tumor progression and response to chemotherapy,suggesting their potential application as prognostic and predictive biomarkers.The role of microRNAs in lung cancer remains elusive.It has been recently reported that epidermal growth factor receptor （EGFR） and hepatocyte growth factor receptor （MET） tyrosine kinase can regulate expression of specific microRNAs including miR-30b,miR-30c,miR-221,miR-222,miR-103 and miR-203,and induce tumorigenesis and gefitinib resistance in lung cancers.We intend to study the role of miR-30b and miR-30c expression in predicting response to tyrosine kinase inhibitors （TKIs） in non-small cell lung cancer （NSCLC）.Methods We have therefore retrospectively examined expression of miR-30b miR-30c in 41 formalin fixed paraffin embedded tissue samples from NSCLC patients when TKIs were used as first line therapy.Results We found a significant correlation between expression of miR-30b and miR-30c.Furthermore,miR-30b and miR-30c expression correlated with short-term response.Kaplan-Meier analysis further revealed that the expression of miR-30b and miR-30c predicted progression free survival and the overall survival rate in the examined cohort.Conclusion Our study identified miR-30b and miR-30c as useful prognostic predictors in NSCLC patients who underwent first line treatment with TKIs.     

A scoring system for prediction of early recurrence after liver resection for Barcelona Clinic Liver Cancer stage B hepatocellular carcinoma

Background The management of Barcelona Clinic Liver Cancer (BCLC) stage B hepatocellular carcinoma (HCC) is controversial due to the early recurrence after curative hepatectomy, and many variables were related to the prognosis. The purpose of this study was to predict the tumor recurrence in early postoperative period of the patients with BCLC stage B HCC.

Methods From January 2004 to January 2012, 104 patients with BCLC stage B HCC underwent hepatectomy. Clinicopathological factors and follow-up data were statistically analyzed to establish a predicting scoring system.

Results The overall survival rates for one, three, and five years were 69.2%, 52.7%, and 42.3%, and the disease-free survival rates for one, three, and five years were 52.9%, 47.3%, and 37.5%, respectively. The multiple factors analysis showed that the micro-vessel invasion, lymph nodes metastasis, multiple lesions, and the high expression of HMGB1 were independent factors (P <0.05). A scoring system was established to predict the early recurrence within one year after the surgery for BCLC stage B HCC, according to the analysis results with a specificity of 85.1% and a sensitivity of 80.3%.

Conclusion Variant clinicopathological factors were associated with early postoperative recurrence for BCLC stage B HCC and recurrence early after hepatectomy was more likely in patients with a higher score of the scoring system.

     

Correlation between molecular biomarkers and risk factors for the clinical progression of benign prostatic hyperplasia using tissue microarray immunostaining

Background The pathogenesis of benign prostatic hyperplasia （BPH） has been widely studied,and several biomarkers are known to play roles in its development.This study aimed to investigate the possible role of cysteine-rich protein 61 （CYR61）,vascular endothelial growth factor （VEGF）,androgen receptor （AR）,interleukin-6 （IL-6）,cytochrome c,caspase-3,and proliferating cell nuclear antigen （PCNA） in the clinical progression of BPH.Methods Tissue specimens from 96 BPH cases who underwent transurethral resection of the prostate were processed and transferred to tissue microarrays.Patient age,prostate volume,serum prostate-specific antigen （PSA） level,and International Prostate Symptom Score （IPSS） of all BPH cases were collected before surgery.The expression of CYR61,VEGF,AR,IL-6,cytochrome c,caspase-3,and PCNA was examined by immunostaining in the BPH specimens,and any possible correlation between the different biomarkers and risk factors for BPH clinical progression was analyzed.Results The expression of CYR61,VEGF,AR,IL-6,cytochrome c,caspase-3,and PCNA in the BPH cases was 68.8％ （66/96）,77.1％ （74/96）,43.8％ （42/96）,31.3％ （30/96）,35.4％ （34/96）,56.3％ （54/96）,and 29.2％ （28/96）,respectively.The expression of both CYR61 and VEGF was positively correlated with patient age,prostate volume,and serum PSA level （P ＜0.05）.Furthermore,cytochrome c and caspase-3 expression were inversely related to prostate volume （P ＜0.05）,and AR expression was positively related to serum PSA level （P ＜0.05）.Conclusion CYR61 and VEGF expression might serve as biomarkers for predicting the clinical progression of BPH due to effects on stromal cell proliferation and angiogenesis.     

Prognostic significance of metastatic lymph nodes ratio in patients with gastric adenocarcinoma after curative gastrectomy

Background We evaluated the impact of the number of metastatic lymph nodes and the metastatic lymph nodes ratio （the ratio between metastatic lymph nodes and total dissected lymph nodes,MLNR） in patients with gastric adenocaminoma following curative gastrectomy and also analyzed the relationship between the number of removed lymph nodes and prognosis in node-negative gastric cancer.Methods From January 2005 to December 2010,1 390 patients who were diagnosed with gastric adenocarcinoma and underwent curative gastrectomy were included.In particular,lymph node metastasis was not present in 515 patients.The number of metastatic lymph nodes and the metastatic lymph nodes ratio were selected for univariate and multivariate analyses to evaluate their influences on the disease outcome.The survival curve was presented according to the number of removed lymph nodes in node-negative gastric cancer using Kaplan-Meier plots.Results The overall 5-year survival rate was 54％ in this group.Univariate analysis revealed that age category,macroscopic appearance,histological grade,tumor size,depth of primary tumor invasion,number of metastatic lymph nodes,metastatic lymph nodes ratio,tumor,nodes,metastasis-classification （TNM） stage and status of lymphovascular,and vessel invasion have significant impact on survival.The number of metastatic lymph nodes and the metastatic lymph nodes ratio both have significant impact on survival （P ＜0.001）.However,in multivariate analyses,only the metastatic lymph nodes ratio was identified to be an independent prognostic factor （P ＜0.001）.The number of removed lymph nodes in node-negative was a strong prognostic factor of survival,the more lymph nodes dissected,the better the survival.Conclusions The metastatic lymph nodes ratio has more significant prognostic value for survival in patients with gastric cancer following curative gastrectomy than the number of metastatic lymph nodes.The number of removed lymph nodes miaht be an important proanostic factor for gastric cancer without lymph node metastasis.     

Role of paxillin in colorectal carcinoma and its relationship to clinicopathological features

Background Colorectal carcinoma is one of the most common malignant tumors.Despite advances in therapy,mortality is still very high.The aim of this study was to evaluate the expression of paxillin in the human colon adenocarcinoma cell line SW480 and its role in cell cycle and apoptosis.We also investigated the expression of paxillin in colorectal carcinoma tissues and its relationship to clinicopathological features and survival.Methods Paxillin short hairpin RNA （shRNA） was constructed and transfected into the colon adenocarcinoma cell line SW480.The influence of paxillin shRNA on the cell cycle and cell apoptosis was analyzed by flow cytometry.Immunohistochemistry staining was used to assess the expression of paxillin and its association with the expression of carcinoembryonic antigen （CEA）,carbohydrate antigen （CA） 19-9,p53 and Bcl-2 in 102 patients with primary colorectal carcinoma.Western blotting was also used to investigate the expression of paxillin.Medical records were reviewed and a clinicopathological analysis was performed.Results In vitro,the percentage of cells in S phase was （45.23±1.05）％,（43.53±1.23）％,and （36.13±0.57）％ in the blank control group,negative control group,and paxillin shRNA group respectively.It was significantly decreased in the paxillin shRNA group （P=0.000）.The early apoptosis index of the paxillin shRNA group （17.2±1.18％） was significantly increased compared to the control shRNA group （（13.17±1.15）％,P=0.013）.Paxillin was positive in 71 （69.6％） patients,and it was found to be overexpressed in tumor tissues compared with normal adjacent tissues.Paxillin positive rate was higher in patients who are less than 50-years old （100.0％ vs.65.6％,P=0.016）.Paxillin expression was associated with a high histologic grade of carcinoma （81.4％ vs.61.0％,P=0.031）,a high rate of regional lymph node metastasis （22.5％ vs.13.0％,P=0.031）,mesenteric artery lymph node metastasis （100.0％ vs.64.8％,P=0.008）,distant metastasis （94.1％ vs.64.7％,P=-0.016） and a high Tumor Node Metastasis （TNM） stage （94.1％,73.2％,60.0％,and 50％,P=0.030）.Multivariate analyses revealed that recurrence was associated with the rate of regional lymph node metastasis （P=0.001） and paxillin expression （P=0.024）.Multivariate analysis indicated that the overall survival is related to the TNM stage （P=0.000）.Conclusions In vitro,paxillin may promote cell proliferation and inhibit apoptosis in SW480 cells.Paxillin may be a potential metastasis predictor,and an independent prognosis factor of recurrence.It may also be related to poor patient outcomes,but was not an independent predictor of survival.     

Effect of neoadjuvant chemotherapy on expressions of estrogen receptor, progesterone receptor, human epidermal growth factor receptor 2, and Ki-67 in breast cancer

Background This study was designed in an attempt to determine the influence of neoadjuvant chemotherapy on estrogen receptor （ER）, progesterone receptor （PR）, human epidermal growth factor receptor 2 （Her-2）, and Ki-67 expressions in patients with breast cancer. Methods Pre- and post-neoadjuvant chemotherapy, paired-tumor specimens from 103 patients with breast cancer administrated with anthracycline or anthracycline combined taxane regimen were collected. Immunohistochemical staining for ER, PR, Her-2, and Ki-67 was performed by the DAKO EnVision method. Results Among the 103 cases, five patients （4.9%） had a complete response （CR）, 82 （79.6%） partial response （PR）, 15 （14.6%） stable disease （SD）, and one （0.9%） progressive disease （PD）, yielding an overall response rate （CR ＋ PR） of 84.5%. Nine patients achieved pathological CR. There was a significant decrease in the average index of Ki-67 post- neoadjuvant chemotherapy, compared with that before chemotherapy （24.1% vs. 39.7%, P 〈0.001）. After neoadjuvant chemotherapy, the changes of Ki-67 in different subtypes of breast cancer were different （P 〈0.001）, and these changes correlated with response to neoadjuvant chemotherapy （P 〈0.001）. No significant changes in immunohistochemical expression were observed for ER, PR and Her-2. Conclusions Neoadjuvant chemotherapy apparently reduced Ki-67 index in primary breast carcinomas, but profiles for ER, PR and Her-2 were not significantly different before and after neoadjuvant chemotherapy. The change of Ki- 67 correlated with molecular subtypes and response to neoadjuvant chemotherapy, suggesting that Ki-67 index was a surrogate marker to predict the treatment response of neoadjuvant chemotherapy.     

43例胃神经内分泌癌的临床特征及根治性手术术后的预后因素分析

Background Gastric neuroendocrine carcinomas （g-NECs） are rare tumors that have aggressive biological behaviors and poor prognosis,but the prognostic factors of postoperative patients with g-NEC are still unclear.Our aim was to study and explore the clinical characteristics and prognostic factors of patients with g-NEC treated with radical surgery.Methods The clinical data of 43 g-NEC patients who underwent surgery from January 2002 to January 2011 at the Zhongshan Hospital of Fudan University were analyzed.Follow-up was conducted by telephone,mail,or returning visit survey.Results The sizes of the 43 neuroendocrine carcinomas （G3） were 1.5 cm × 1.5 cm × 0.5 cm to 7 cm × 8 cm × 1.5 cm.Eight NECs were localized,and 35 had lymph node involvement,of which 1 also had hepatic metastasis.At the end of the follow-up,the follow-up rate was 97.7％ （42/43）,and the median follow-up time was 22.2 months.The median overall survival of g-NEC patients was 36.5 months,and the 1-,3-,and 5-year overall survival rates were 86.0％,51.6％,and 36.7％,respectively.Sex （P ＜0.05） and lymph node involvement （P ＜0.05） were prognostic factors of postoperative g-NEC patients,among which sex was an independent prognostic factor （P ＜0.05）,as a survival advantage of female patients over male was observed.Conclusions Most of the g-NECs were diagnosed at an advanced stage.The prognosis of g-NECs was related with sex and lymph node involvement,of which sex was an independent prognostic factor,with female patients having a survival advantage.     

Retrospective study of mycophenolate mofetil treatment in IgA nephropathy with proliferative pathological phenotype

Background Mycophenolate mofetil （MMF） and cyclophosphamide （CTX） are widely used in treating various kidney diseases.However,whether they are effective and which one is better for treating IgA nephropathy patients with proliferative pathological phenotype in renal diseases,such as endocapillary proliferation,cellular crescents,and/or capillary loops fibrinoid necrosis is still unknown.We,therefore,initiated a study to compare the effects of MMF and CTX in treating IgA nephropathy with the above pathological lesions.Methods One hundred and nineteen patients with IgA nephropathy who had at least one of the three aforementioned lesions were enrolled.All patients were treated with prednisone; 48 patients received prednisone only （Pred group）,40 received MMF and prednisone （MMF ＋ Pred group）,and 31 were treated with CTX and prednisone （CTX ＋ Pred group）.The median time of follow-up was 30 months （maximum：96 months）.The primary endpoint was defined as renal survival.The incidence of remission of proteinuria was the secondary endpoint.Results Serum creatinine in all groups declined significantly at different follow-up times （P=0.002）,and the differences among the three groups were significant （P＜0.001）.At 24 months of follow-up,the decline rates were 12.35％,32.95％,and 24.14％ in the Pred,MMF ＋ Pred,and CTX ＋ Pred groups respectively.For urine protein excretion,the decline rates were 49.12％ （Pred）,73.67％ （MMF ＋ Pred）,and 63.53％ （CTX ＋ Pred） respectively at 24 months of follow-up.The differences among the three groups were not significant （P=0.714）.Renal survival （the primary endpoint） was significantly different （P=0.027）; however,the sencondary endpoint was similar for all the three groups （P=0.100）.Conclusions For IgA nephropathy patients with endocapillary proliferation,cellular crescents,and/or fibrinoid necrosis of capillary loops,prednisone combined with MMF was more effective in lowering the serum creatinine than with CTX.Combined MMF and orednisone treatment led to a better renal survival compared to that of prednisone with CTX.     

Predictive factors improving survival after gastric and hepatic surgical treatment in gastric cancer patients with synchronous liver metastases

Background  The prognosis for patients with gastric cancer and synchronous liver metastases is very poor. However, a standard therapeutic strategy has not been well established. The clinical benefit and prognostic factors after hepatic surgical treatment for liver metastases from gastric cancer remain controversial.

Methods  Records of 105 patients who underwent gastrectomy regardless of hepatic surgical treatment for gastric cancer with synchronous liver-only metastases in our center between 1995 and 2010 were retrospectively reviewed.

Results  The overall survival rate for the 105 patients was 42.1%, 17.2%, and 10.6% at 1, 2, and 3 years, respectively, with a median survival time of 11 months. Multivariate survival analysis revealed that the extent of lymphadenectomy (D) (P <0.001), lymph node metastases (P <0.001), extent of liver metastases (H) (P=0.008), and lymphovascular invasion (P=0.002) were significant independent prognostic factors for survival. Among patients who underwent D2 lymphadenectomy, those who underwent hepatic surgical treatment had a significantly improved survival compared with those who underwent gastrectomy alone (median survival, 24 vs. 12 months; P <0.001). However, hepatic surgical treatment was not a prognostic factor for patients who underwent D1 lymphadenectomy (median survival, 8 vs. 8 months; P=0.495). For the 35 patients who underwent gastrectomy plus hepatic surgical treatment, D2 lymphadenectomy (P <0.001), lymph node metastases (P=0.015), and extent of liver metastases (H1 vs. H2 and H3) (P=0.017) were independent significant prognostic factors for survival.

Conclusions  D2 lymphadenectomy plus hepatic surgical treatment may provide hope for long-term survival of judiciously selected patients with hepatic metastases from gastric cancer. Patients with a low degree of lymph node metastases and H1 liver metastases would make the most appropriate candidates. However, if D2 dissection cannot be achieved, hepatic surgical treatment is not recommended.

     

Integrated positron emission tomography and computed tomography in preoperative lymph node staging of non-small cell lung cancer

Background Integrated positron emission tomography and computed tomography （PET/CT） is increasingly used for the preoperative nodal staging of non-smail cell lung cancer （NSCLC）.The aim of this study was to evaluate the accuracy of PET/CT in comparison with CT in detection of nodal metastasis and preoperative nodal staging in patients with NSCLC,and to analyze the causes of the PET/CT false-negative and false-positive results.Methods Consecutive patients with pathologically proven NSCLC who underwent staging using PET/CT from July 2008 to February 2012 were evaluated retrospectively.Nodal staging was pathologically confirmed on tissue specimens obtained at thoracotomy.The accuracy of PET/CT and CT in the assessment of intrathoracic nodal involvement was determined using histological results as the reference standard.Logistic regression was used to define the causes of the false-negative and false-positive results.Results A total of 528 lymph node stations were evaluated in 101 patients.Lymph nodes were positive for malignancy in 43 out of 101 patients （42.6%）,and 101 out of 528 nodal stations （19.2%）.PET/CT was significantly more accurate for nodal staging than CT.The sensitivity,specificity,positive and negative predictive values,and accuracy of PET/CT for detecting nodal metastasis were 51.5%,95.8%,74.3%,89.3%,and 87.3% and the corresponding data by CT were 45.5%,87.1%,45.5%,87.1%,and 79.2%,respectively.PET/CT confers significantly higher specificity,positive predictive value,and accuracy than CT in detecting nodal metastasis.False-negative results by PET/CT are significantly associated with smaller lymph node size,whereas false-positive results are related to a combination of inflammatory disorders and larger lymph node size.Conclusion PET/CT confers significantly higher accuracy than CT in nodal staging,and is more specific and accurate than CT in detecting nodal metastasis but has a low sensitivity and high false-negative rate.     

Urinary albumin excretion rate: a risk factor for retinal hard exudates in macular region in type 2 diabetic patients

Background The various risk factors for retinal hard exudates are still poorly understood in type 2 diabetic patients.The aim of this study was to determine the association between urinary albumin excretion rate （UAER） and hard exudates in macular region in north Chinese patients.Methods A total of 272 patients （272 eyes） were enrolled for this study,including 154 subjects from group 1 （mild hard exudates）,91 subjects from group 2 （moderate hard exudates） and 27 subjects from group 3 （severe hard exudates） confirmed using colour fundus photography,optical coherence tomography （OCT） as well as slit-lamp biomicroscopy with 78 diopter （D） lens.Each participant underwent a comprehensive assessment that included biochemical,clinical characteristics test and detailed ophthalmic evaluation.One-way analysis of variance （ANOVA） test and chi-square test were performed to analyze the fasting blood glucose （FBG）,glycated hemoglobin （HbA1c）,total cholesterol （TC）,low density lipoprotein （LDL）,high density lipoprotein （HDL）,triglycerides （TG）,full blood counts,urinary albumin excretion rate （UAER）,blood creatinine （CREA）,duration of diabetes,body mass index （BMI）,systolic blood pressures （SBP） and diastolic blood pressures （DBP） between groups.Ordinal logistic regression analysis was further performed in order to eliminating the possible confounding factors.Results Three groups were matched in terms of age and gender.Risk factors which showed significant difference between groups include FBG （P ＜0.001）,HbA1c （P ＜0.001）,LDL （P ＜0.001）,UAER （P ＜0.001）,duration of diabetes （P=0.001）,TC （P=0.005）,SBP （P=0.026）,CREA （P=0.004） and haemoglobin （Hb） （P=0.012）.There was no significant difference between groups for the TG,HDL,DBP,platelet,total white blood cells and BMI.Using ordinal Logistic regression analyses,of all the variables,HbA1c,LDL and UAER which were independent risk factor for hard exudates showed a si     

Silencing of osteopontin promotes the radiosensitivity of breast cancer cells by reducing the expression of hypoxia inducible factor 1 and vascular endothelial growth factor

Background  Osteopontin (OPN) is a secreted phosphoglycoprotein (SSP) that is overexpressed in a variety of tumors and was regarded as a molecular marker of tumors. In this study, we intended to demonstrate the role of OPN in human breast cancer cell line MDA-MB-231.

Methods  Recombinant plasmid expressing small interfering RNA (siRNA) specific to OPN mRNA was transfected into MDA-MB-231 cells to generate the stable transfected cell line MDA-MB-343, and the empty plasmid tansfected cells (MDA-MB-neg) or wildtype MDA-MB-231 cells were used as control cells respectively. Expression of OPN, hypoxia inducible factor-1 (HIF-1) and vascular endothelial growth factor (VEGF) proteins was analyzed by Western blotting analysis. The radiosensitivity of cells was determined by detecting cell apoptosis, cell proliferation and cell senescence.

Results  HIF-1 and VEGF proteins in MDA-MB-343 cells were significantly downregulated upon the efficient knockdown of OPN expression under either hypoxia or normoxia environment. Moreover, expression of OPN protein was upregualted upon hypoxic culture. Stable OPN-silencing also decreased cell invasion, increased cell apoptosis and cell senescence, as well as reduced clonogenic survival, resulting in increase radiation tolerance.

Conclusions  Suppression of OPN gene expression can enhance radiosensitivity and affect cell apoptosis in breast cancer cells. OPN seems to be an attractive target for the improvement of radiotherapy.

     

Blockade of the OX40/OX40L pathway and induction of PD-L1 synergistically protects mouse islet allografts from rejection

Background OX40/OX40 ligand （OX40/OX40L） and programmed death-1/programmed death ligand-1 （PD-1/PD-L1） co- stimulator/signals play important roles in T cell-induced immune responses. The aim of this study was to investigate the roles of OX40/OX40L and PD-1/PD-L1 costimulatory pathways in mouse islet allograft rejection. Methods Lentiviral vectors containing OX40L siRNA sequences and an adenovirus vector containing the PD-L1 gene were constructed. The streptozotocin-induced model of diabetes was established in C57BL/6 （H-2b） mice. Diabetic C57BL/6 mice were randomly allocated into five groups： group 1, untreated control; group 2, Ad-EGFP treatment; group 3, Ad-PD-L1 treatment; group 4, OX40L-RNAi-LV treatment; group 5, OX40L-RNAi-LV combined with Ad-PD-L1 treatment. Lentiviral vector and the adenovirus vector were injected, singly or combined, into the caudal vein one day before islet transplantation. The islets of DBA/2 （H-2d） mice were transplanted into the renal subcapsular space of the diabetic recipients. Recipient blood glucose and the survival time of the allografts were monitored. Antigen-specific mixed lymphocyte reaction was also evaluated.