MicroRNA 145 may play an important role in uveal melanoma cell growth by potentially targeting insulin receptor substrate-1

Background MicroRNAs （miRNAs） contribute to tumorigenesis by acting as either oncogenes or tumor suppressor genes. In this study, we investigated the role of miR-145 in the pathogenesis of uveal melanoma. Methods Expression profiles of miRNAs in uveal melanoma were performed using Agilent miRNA array. Quantitative real-time polymerase chain reaction was used to screen the expression levels of miR-145 in normal uveal tissue, uveal melanoma tissue, and uveal melanoma cell lines. Lenti-virus expression system was used to construct MUM-2B and OCM-1 cell lines with stable overexpression of miR-145. Cell proliferation, cell cycle, and cell apoptosis of these miR-145 overexpression cell lines were examined by MTT assay and flow cytometry respectively. The target genes of miR-145 were predicted by bioinformatics and confirmed using a luciferase reporter assay. The expression of insulin-like growth factor-1 receptor （IGF-1R）, insulin receptor substrate-1 （IRS-1） proteins was determined by Western blotting analysis. IRS- 1 was knocked down in OCM-1 cells. TUNEL, BrdU, and flow cytometry assay were performed in IRS-1 knocked down OCM-1 cell lines to analyze its function. Results Forty-seven miRNAs were up regulated in uveal melanoma and 61 were down regulated, miR-145 expression was significantly lower in uveal melanoma sample and the cell lines were compared with normal uveal sample. Overexpression of miR-145 suppressed cell proliferation by blocking the G1 phase entering S phase in uveal melanoma cells, and promoted uveal melanoma cell apoptosis. IRS-1 was identified as a potential target of miR-145 by dual luciferase reporter assay. Knocking down of IRS-1 had similar effect as overexpression of miR-145. Conclusion miR-145 might act as a tumor suppressor in uveal melanoma, and downregulation of the target IRS-1 might be a potential mechanism.     

MicroRNAs are implicated in the initiation and progression of gastric cancer

Objective Gastric cancer is a genetically heterogeneous disease that progresses via different oncogenes.MicroRNA （miRNA） can regulate oncogene expression at the post-translational level.In this review,we summarize the most commonly altered miRNAs and their possible roles in cancer initiation and progression in gastric cancer.Data sources Most articles were identified by searching PubMed online resources using the key terms of microRNA and gastric cancer.Study selection Mainly original milestone articles and critical reviews written by major pioneer investigators in the field were selected,and the 69 most important articles were cited finally.Results A set of miRNAs are consistently deregulated in gastric cancer,although there is no clear miRNA expression profiles,such as miR-21 and miR-17 （～92 clusters）.These deregulated miRNAs play important roles in promoting cell proliferation,tumor metastasis,and chemotherapeutic resistance in gastric cancer by targeting different oncogenes.Clinical relevance of these deregulated miRNAs is proved to be associated with TNM stages,metastasis,and prognosis of gastric cancer patients.In addition,circulating miRNAs are promising noninvasive biomarkers for gastric cancer.Conclusions miRNAs have produced a novel paradigm in research in gastric cancer.These small molecules play macroroles in gastric cancer initiation and progression.These results will help us improve management of gastric cancer in future.     

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.     

Pioglitazone inhibits the expression of nicotinamide adenine dinucleotide phosphate oxidase and p38 mitogen-activated protein kinase in rat mesangial cells

Background Oxidative Stress and p38 mitogen-activated protein kinase （p38MAPK） play a vital role in renal fibrosis. Pioglitazone can protect kidney but the underlying mechanisms are less clear. The purpose of this study was to investigate the effect of pioglitazone on oxidative stress and whether the severity of oxidative stress was associated with the phosphorylation level of p38MAPK.     

Duodenal-jejunal bypass surgery on type 2 diabetic rats reduces the expression of matrix metalloproteinase-9 and tissue inhibitor of matrix metalloproteinase-1 in the thoracic aorta

Background Bariatric surgery offers a productive resolution of type 2 diabetes mellitus （T2DM）.The development of T2DM vasculopathy is due to chronic inflammation,which increases matrix metalloproteinase-9 （MMP-9） and tissue inhibitor of matrix metalloproteinase-1 （TIMP-1） expression.This study sought to examine MMP-9 and TIMP-1 expression in the thoracic aorta after duodenal-jejunal bypass （DJB） surgery on a T2DM rat model induced by a high-fat diet and low dose streptozotocin （STZ）.Methods Twenty-one T2DM Wistar rats induced by high-fat diet and low dose STZ were randomly divided into DJB and sham duodenal-jejunal bypass （S-DJB） groups.Ten Wistar rats were fed a normal diet as a control.Recovery of gastrointestinal function post-operation and resumption of a normal diet completed the experiment.Body weight,blood glucose,blood lipid levels,and MMP-9 and TIMP-1 expression levels in aortic endothelial cells were measured throughout.Results DJB rats showed significant weight loss 2 weeks post-operation compared with S-DJB rats.After surgery,DJB rats showed significant improvement and steady glycemic control with improved insulin sensitivity and glucose tolerance.They also exhibited improved lipid metabolism with a decrease in fasting free fatty acids （FFAs） and triglycerides （all P ＜0.05）.Immunohistochemistry showed decreased MMP-9 and TIMP-1 expression 12 weeks after surgery （P ＜ 0.01）.Conclusions DJB surgery on an induced T2DM rat model improves blood glucose levels and lipids,following a high-fat diet and low dose STZ treatment.In addition,DJB decreased MMP-9 and TIMP-1 expression in vascular endothelial cells,which may play an important role in delaying the development of T2DM vascular disease.     

Changes in dendritic cells and dendritic cell subpopulations in peripheral blood of recipients during acute rejection after kidney transplantation

Background Advances in transplantation immunology show that the balance between dendritic cells （DCs） and their subsets can maintain stable immune status in the induction of tolerance after transplantation. The aim of this study was to investigate if DCs and DC subpopulations in recipient peripheral blood are effective diagnostic indicators of acute rejection following kidney transplantation. Methods Immunofluorescent flow cytometry was used to classify white blood cells （WBCs）, the levels of mononuclear cells and DCs （including the dominant subpopulations, plasmacytoid DC （pDC） and myeloid DC （mDC）） in peripheral blood at 0, 1, 7, and 28 days and 1 year after kidney transplantation in 33 patients. In addition, the blood levels of interleukin-10 （IL-10） and IL-12 were monitored before and after surgery. Fifteen healthy volunteers served as normal controls. Patients were undertaking hemodialysis owing to uremia before surgery. Results The total number of DCs, pDC, and mDC in peripheral blood and the pDC/mDC ratio were significantly lower in patients than controls （P 〈0.05）. Peripheral DCs suddenly decreased at the end of day 1, then gradually increased through day 28 but remained below normal levels. After 1 year, levels were higher than before surgery but lower than normal. The mDC levels were higher in patients with acute rejection before and 1 day after surgery （P 〈0.005）. There was no significant difference in IL-10 and IL-12 levels between patients with and without acute rejection. Conclusion The changes in DCs and DC subpopulations during the acute rejection period may serve as effective markers and referral indices for monitoring the immune state, and predicting rejection and reasonably adjusting immunosuppressants.     

Tartrate-resistant acid phosphatase 5b is a potential biomarker for rheumatoid arthritis: a pilot study in Han Chinese

Background Bone damage around the joints is one of the major pathophysiological mechanisms that leads to rheumatoid arthritis （RA） chronic disability.Serum tartrate-resistant acid phosphatase 5b （TRACP-5b） is secreted by osteoclasts,its activity can be used as a clinically relevant bone resorption marker.The aim of this study was to test whether the measurement of serum levels of TRACP-5b in patients with RA would correlate with measures of disease activity and with responses to therapy.Methods Fifty-six patients were randomly assigned to receive recombinant human cytotoxic tlymphocyte-associated antigen-4 immunoglobulin （RhCTLA4-lg）,infliximab or methotrexate （MTX）.The clinical and serologic indicators of RA activity were evaluated at baseline and at 24 weeks.Serum TRACP-5b was measured by Enzyme-linked Immunosorbent Assay （ELISA） at 0,12 and 24 weeks.Hand X-rays were obtained at baseline.Results At baseline,the levels of TRACP-5b correlated with the severity of X-ray damage,disease duration （r=0.332,P=0.012）,and tender joint count （r=0.408,P=0.002）.The 24 weeks values of TRACP-5b for RhCTLA4-lg group and infliximab group differed significantly from the baseline values in each group （P 〈0.05; P 〈0.05）,whereas only the value for RhCTLA4-lg group differed significantly from the 24 weeks value for the MTX group （P 〈0.01）.Considering the two biologics-treated groups together,the TRACP-5b levels at 24 weeks differed significantly from the baseline values only in those patients who reached an ACR70 level （P 〈0.05）.Conclusions Measurement of serum TRACP-5b in RA patients reflects clinical and radiological measures of disease activity,treatment with certain biologics,and degree of response to therapy.TRACP-5b should be investigated further as a potential biomarker to predict response to therapy,including slowing of radiographic progression.     

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.     

Kinetics of serum HBsAg in Chinese patients with chronic HBV infection with long-term adefovir dipivoxil treatment

Background Knowledge on Hepatitis B surface antigen （HBsAg） kinetics in chronic hepatitis B （CHB） patients with longterm adefovir dipivoxil （ADV） treatment is limited.The aims of this study were to investigate HBsAg kinetics in patients with chronic hepatitis B virus （HBV） infection treated with long-term ADV and to evaluate different characteristics between patients with and without HBsAg loss.Methods We retrospectively evaluated HBsAg kinetics in 24 Chinese patients with chronic HBV infection who achieved continuous virologic suppression during ADV therapy.HBV genotype was determined at baseline.Liver biochemistry,hepatitis B e antigen status,serum HBV DNA,and HBsAg levels were measured at baseline,6 months,and once every year thereafter.Results Of these 24 patients,3,1,and 20 patients were followed up for 3,5,and 6 years,respectively.Baseline serum HBsAg level had a moderate correlation with baseline HBV DNA level （r=0.52,P=0.01）.The median rate of HBsAg reduction during the therapy period was 0.08 Ig IU·ml-1·y-1.Baseline serum HBsAg level was significantly higher than other time points （P ranges from 0.046 to 0.002）.The HBsAg reduction rate during the first year was similar to that in other years （P〉0.05）.The HBsAg reduction rate during the first year in patients with eventual HBsAg loss was significantly faster than that in patients without HBsAg loss （P=0.005）.Conclusions Serum HBsAg levels in Chinese CHB patients receiving long-term ADV demonstrated a gradual reduction.Patients with eventual HBsAg loss had a significantly faster HBsAg reduction rate during the first year than those without HBsAg loss.     

Effects of lead exposure on placental cellular apoptosis and endoplasmic reticulum stress in rats

Background Lead exposure during pregnancy contributes to fetal abortion and/or teratogenesis.Endoplasmic reticulum （ER） apoptosis can be induced by various pathological conditions when ER function is disturbed.However,it is unclear whether ER stress and apoptosis play a role in the etiology of lead-exposed disease status.We aimed to investigate whether lead induced placental apoptosis and subsequent toxicity is initiated by ER apoptosis via caspase-12.Methods Sixty-three female Wistar rats were exposed to lead in drinking water during various gestational periods.Blood lead level was determined by atomic absorption spectrophotometry.Placental cytoplasmic organelles were examined by electronic microscopy.Placental caspase-12 mRNA expression was evaluated by qRT-PCR.TUNEL assay was used to determine the placental apoptosis.Results Lead exposure significant induced ER apoptosis compared to that of the controls （P ＜0.05）,accompanied with increased caspase-12 mRNA expression.Significant differences of caspase-12 mRNA expression levels were observed among the four groups （F=13.78,P ＜0.05）.Apoptotic index （AI） was significantly increased in experimental groups compared to that of the controls （F=96.15,P ＜0.05）.In lead-exposed groups,trophoblast cells underwent degeneration and fibrin deposition; Mitochondria were swollen and decreased in number; ER swelling,expansion,and vacuolization were observed.Conclusion Lead exposure contributes to placental apoptosis,as well as increased caspase-12 mRNA expression,which in turn promoted ER stress.     

Overexpression of interleukin-l7 in tumor-associated macrophages is correlated with the differentiation and angiogenesis of laryngeal squamous cell carcinoma

Background  Interleukin-l7 (IL-17), which exerts strong pro-inflammatory effects, has emerged as an important mediator in inflammation-associated cancer. The aim of this study was to clarify the relationship between IL-17 and tumor associated macrophages (TAMs), and the correlation of the microvessel density in the development of laryngeal squamous cell carcinoma (LSCC).

Methods  Histopathological observations and immunohistochemistry staining for IL-17, CD68, and CD34 were performed on 72 specimens (32 cases of LSCC, 20 cases of adjacent tissues of carcinoma as controls, and 20 cases of chronic hypertrophic laryngitis). Double immunohistochemical staining was done to determine which cells expressed IL-17. Real-time quantitative PCR determined the mRNA expression of IL-17. ELISA was used to detect the expression of the serum level of IL-17 in the three groups.

Results  The inflammation response had increased in LSCC. Overexpression of IL-17 and CD68 protein were seen in LSCC (P <0.01). The expression of IL-17 was different between well and poorly differentiated LSCC (P <0.01). The IL-17 expressing cells were mainly located in macrophages (CD68⁺/IL17⁺) as demonstrated by double immunohistochemical staining. IL-17 expression significantly correlated with high microvessel density (CD34⁺) in LSCC (P <0.05). Relatively higher mRNA expression levels of IL-17 were seen in LSCC compared to the controls (P <0.05). The serum expression of IL-17 was similar among the three groups (P >0.05).

Conclusion  IL-17 was expressed by TAMs, and IL-17 may significantly correlate to the differentiation and angiogenesis in the development of LSCC.

     

Heliox as a driving gas to atomize inhaled drugs on acute exacerbation of chronic obstructive pulmonary disease: a prospective clinical study

Background Acute exacerbation of chronic obstructive pulmonary disease （AECOPD） is a common condition,which affects not only the quality of life of patients but also their prognosis.The purpose of this study was to explore the effects of an inhaled salbutamol sulfate solution and an inhalation suspension of the glucocorticoid budesonide that were atomized with heliox to treat patients with AECOPD.Methods Twenty-three patients with AECOPD were divided into a treatment group （He/O2=70％/30％） and a control group （N2/O2=70％/30％）.The salbutamol sulfate and budesonide were administered by inhalation twice a day for 7 days.Vital signs,arterial blood gas levels,pulmonary function and the levels of serum myostatin （sMSTN） were measured and lung vibration imaging was performed.Results We found that the PaO2 and PaCO2 values were not significantly different between the two groups at the various time points （P ＞0.05）.There were also no significant differences in any of the parameters of pulmonary function between the two groups.However,after baseline correction,the increase rate of the forced expiratory volume in one second （FEV1）,the forced vital capacity （FVC）,and the maximum minute ventilation （MW） appeared to be significantly increased at some time points compared with the baseline （before treatment） in both groups （P ＜0.05）.Although the values of quantitative lung distribution （QLD） for different regions and the levels of sMSTN were slightly different between the two groups,the repeated measures analysis of variance （ANOVA） revealed that there were no significant differences between the two groups or within any group （P ＞0.05）.Conclusion Although the use of heliox as a driving gas can improve symptoms and benefit patients with AECOPD,the heliox treatment group did not have significant differences in arterial blood gases,lung function,lung vibration response imaging or the levels of sMSTN compared with the control group.（Chinese Clinical Trial Register Center ChiCTR-TRC-00000273）     

Intermittent hypoxia with or without hypercapnia is associated with tumorigenesis by decreasing the expression of brain derived neurotrophic factor and miR-34a in rats

Background Very recent studies revealed that obstructive sleep apnoea （OSA） is a contributor of the increased incidence and mortality of cancer in humans,but mechanisms of how OSA promotes tumorigenesis remains largely unknown.We investigated whether intermittent hypoxia with and without hypercapnia plays a role in tumorigenesis.Methods First,Sprague-Dawley （SD） male rats （12 weeks old） were subjected to different hypoxia exposures：intermittent hypoxia and intermittent hypoxia with hypercapnia; continuous hypoxia and normal air.The systemic application of chronic fast rate hypoxia with or without hypercapnia mimicked severe OSA patients with apnoea/hypopnea index equivalent to 60 events per hour.Then routine blood tests were performed and the levels of brain derived neurotrophic factor （BDNF） and miR-34a were examined.Results In contrast to intermittent hypoxia with hypercapnia,both intermittent hypoxia and continuous hypoxia treatments caused significantly higher levels of haematology parameters than normoxia treatments.Compared to normoxia,intermittent hypoxia with hypercapnia exposure resulted in substantial decrease of serum BDNF and,miR-34a in the lower brainstem,while less pronounced results were found in intermittent hypoxia and continuous hypoxia exposure.Conclusions The exposure of intermittent hypoxia with or without hypercapnia,mimicking the situations in severe OSA patients,was associated with,or even promoted tumorigenesis.     

Effect and mechanism of tacrolimus on melanogenesis on A375 human melanoma cells

Background Topical tacrolimus has been used for vitiligo as a common treatment option for more than ten years while the underlying mechanism is still uncertain.The aim of this study was to investigate the direct effects of tacrolimus on the melanogenesis and migration on human A375 melanoma cells.The expression of c-KIT mRNA and protein of human A375 cells were also investigated.Methods The cultured A375 human melanoma cells were randomly assigned to control and tacrolimus treatment groups （10,102,103and 104 nmol/L）.The cell proliferation was measured with Cell Counting Kit-8 assays.Melanin content was measured with NaOH method.Transwell migration assay was used to measure cell migration.The expression of c-KIT mRNA and protein were measured with real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry respectively.Results The cell proliferation of the 103 and 104 nmol/L tacrolimus groups were significantly lower （0.666±0.062 and 0.496±0.038） as compared with the control （0.841±0.110,P 〈0.05）.The mean melanin content in all groups treated with different concentration of tacrolimus （10,102,103,104 nmol/L） increased compared with the control group （P 〈0.05）.Dosedependent increase in cell migration were seen in all tacrolimus-treated groups （P 〈0.01）.The expression of c-KIT mRNA level in A375 cells exposed to tacrolimus （103and 104 nmol/L） had significantly increased by 3.03-fold and 3.19-fold respectively compared with the control （P 〈0.05）.Conclusions Although tacrolimus had no effects on cell proliferation on A375 human melanoma cells,it could increase the melanin content and cell migration.The expression of c-KIT mRNA and protein increased dose-dependently in tacrolimus-treated groups as compared with the control.Our study demonstrated that tacrolimus could enhance the melanogenesis and cell migration on A375 cells.     

Evaluation of efficacy and safety of sunitinib regimen in 22 patients with metastatic renal cell carcinoma: at least 12-month follow-up

Background Sunitinib has been proved an effective new option for treatment of metastatic renal cell carcinoma (mRCC). Analysis of clinical data of 22 patients, who were exposed to sunitinib for at least 1 year, was conducted to evaluate the long-term efficacy and safety of sunitinib for the treatment of mRCC.

Methods A total of 54 patients with mRCC were treated with sunitinib malate, 50 mg/d orally, on a 4-weeks-on and 2-weeks-off dosing schedule in Peking University First Hospital. Treatment continued until disease progression, unacceptable adverse events (AEs), or death. Among them, 22 patients continued treatment for at least 1 year. The clinical data of these 22 patients were prospectively collected for analysis. AEs were assessed according to the National Cancer Institute Common Terminology Criteria for Adverse Events, Version 3.0. Tumor response was evaluated in accordance with the Response Evaluation Criteria in Solid Tumors.

Results Median progression-free survival was 19.5 months until last follow-up. The best efficacy results achieved were complete response, partial response, and stable disease for 2, 9, and 11 patients, respectively. Objective response rate was 50%. The most common AEs were hand-foot syndrome (95%) and hypertension (91%). Other common AEs were thyroid-stimulating hormone elevation (82%), platelet decrease (77%), and loss of appetite (77%). Only one patient withdrew from treatment for cardiac infarction. Another nine patients experienced dose modifications or short-term suspensions.

Conclusion Long-term exposure to sunitinib malate showed encouraging efficacy in the treatment of mRCC. At the same time, the tolerability was good.