ABSTRACT
Objective The aim of this study was to investigate the role and mechanism of ABL2 in lung cancer and its mech-anism. Methods The expression of ABL2 in lung cancer and adjacent tissues was detected by Real-Time PCR. A lung adenocarci-noma A549 cell line stably expressing of ABL2 was established,and the changes of cell proliferation and migration ability were detec-ted by MTT,cell migration and colony formation assays. Western blot was used to detect the expression of EMT,apoptosis and PI3K/AKT signaling pathway-related proteins. Results The expression of ABL2 in lung cancer tissues was significantly higher than that in adjacent tissues(P<0. 001). After silencing ABL2 in the A549 cells,compared with the control group,the migration ability of cells was weakened after 48 hours(P<0. 001),the growth rate of cells began to slow down from the third day(P<0. 05),and the average number of clones formed after 15 days also decreased(P<0. 01). The expression of E-cadherin( P<0. 001) was increased in the epithelial cell marker after silencing ABL2,and the expression of stromal cell markers N -cadherin ( P <0. 001),Vimentin ( P <0. 01)and Snail(P<0. 001)was decreased. The expression of apoptosis-related protein Bcl-XL(P<0. 01)was decreased and BAX ( P<0. 001)expression was up-regulated. The expression of PI3K/AKT signaling pathway-associated proteins such as PI3K P110 (P<0. 05),AKT(P<0. 01) and p-AKT( P<0. 05) was significantly decreased. Conclusion Silencing ABL2 gene can promote apoptosis,and inhibit proliferation and migration of lung cancer cells through a PI3K/AKT signaling pathway.
ABSTRACT
Objective@#To establish a diagnostic prediction model for esophageal squamous cell carcinoma (ESCC) and search the potential biomarkers of ESCC. @*Methods@#Serum samples from 59 patients with ESCC and 57 healthy controls were collected, and randomly divided into the training group (44 patients and 42 healthy controls) and validation group (15 patients and 15 healthy controls). Serum proteins/peptides were extracted and purified with weak cation-exchange chromatography Magnetic Beads (WCX-MB), and detected by the matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). Then the differentially expressed proteins/peptides were screened out, and a diagnostic prediction model for ESCC was established and preliminarily validated. @*Results@#The ClinProTools software identified 31 differential peptide peaks (P＜0.05), among which 18 peaks had significant difference (P＜0.01). Compared with healthy controls, 8 peaks were up-regulated in ESCC patients, while 10 peaks were down-regulated. Among them, the areas under the receiver operating characteristics (ROC) curve (AUC ROC ) of m/z 2 660.84 and m/z 5 336.49 peaks were 0.95 and 0.91, respectively, and their expressions were up-regulated in ESCC patients. The validation results showed that the accuracy, sensitivity and specificity of the diagnostic prediction model established by the genetic algorithm (GA) were 93.10%, 92.90% and 93.30%, respectively. @*Conclusion@#The established diagnostic prediction model may be used for the auxiliary diagnosis of ESCC. Two peptide peaks of m/z 2 660.84 and m/z 5 336.49 may be the potential biomarkers of ESCC.
ABSTRACT
Objective To establish a rapid diagnostic method for the detection of marine vibrios,and then construct a new technology platform for the clinical diagnosis of marine vibrio infection.Methods A pair of PCR primers and a sequencing primer based on the whA gene of V.vulnificus and the toxR genes of V.parahemolyticus and V.alginolyticus were designed respectively,and then the specific DNA fragments were amplified.Next,the single-stranded DNA templates were prepared for pyrosequencing.The obtained base sequence was validated by NCBI alignment.In addition,the 16S rRNA genotyping of V.vulnificus was also performed.Results The PCR primers and sequencing primer of V.vulnificus showed good specificity,and a 167 bp DNA fragment was amplified from 4 strains of V.vulnificus.The pyrosequencing results completely matched with the whA gene sequence of V.vulnificus.Meanwhile,the control strains were negative.A 105 bp DNA fragment and a 134 bp DNA fragment were amplified from 11 strains of V.parahemolyticus and V.alginolyticus,respectively,and the pyrosequencing results were consistent with the expected sequence.In addition,one of 4 strains of V.vulnificus was identified as 16S rRNA-A type,and the other 3 as 16S rRNA-B type.Conclusion The PCR-pyrosequencing method established in this study is a new method for the real-time detection of short nucleotide sequences.It has some advantages such as high throughput,high precision and simple operation,and may be applied to the fast and accurate identification of marine and terrestrial pathogenic bacteria.
ABSTRACT
Objective: To investigate the effect of different doses of perindopril on peripheral endothelial progenitor cells (EPCs) and vascular endothelial function in patients with coronary artery disease (CAD) . Methods: A total of 84 CAD patients with coronary angiography confirmed diagnosis were divided into 3 groups: Control group, the patients received routine medication, n=27. Low-dose group, the patients received routine medication with perindopril for 4mg, n=29. High-dose group, the patients received routine medication with perindopril for 8mg, n=28. All patients were treated for 12 weeks. The EPCs level was detected by flow cytometry assay, flow-mediated-dilation (FMD) function in brachial artery was measured by ultrasound and plasma levels of high sensitivity C-reactive protein (hs-CRP), angiotensin II (AngII) were examined in all groups. Results: ① After12 weeks of treatment, the EPCs level and FMD function had certain improvement, hs-CRP level decreased in various degrees in all 3 groups, P group showed increased EPCs level and FMD function, decreased levels of hs-CRP and AgnII, P Conclusion: Perindopril may mobilize peripheral EPCs at certain point, and therefore improve endothelial function, the higher dose of perindopril may have better effect.
ABSTRACT
Objective To investigate the effect of PPARγ in acute coronary syndrome (ACS) patients with type 2 diabetes (T2DM) for the severity of coronary atherosclerosis and plaque stability. Methods We selected 102 patients with ACS, including 52 patients with type 2 diabetes mellitus (ACS+T2DM group) and 50 patients with simply ACS (ACS group). Meanwhile, we selected 30 patients without coronary heart disease and T2DM as the control group. All basic clinic data, CAG and the Gensini score were compared among all groups. To all patients, blood was drew when they were enrolled to detect the level of PPARγ and MMP-9. Results Gensini points in the ACS+T2DM group was much higher than that of the ACS group (P < 0.05). The levels of PPARγ of the ACS group and the ACS+T2DM group, when compared with the control group, were decreased significantly, but the level of MMP-9 were increased (all P < 0.05). The level of PPARγ in the ACS+T2DM group was much lower than the ACS group, and the level of MMP-9 was much higher (P<0.05). Gensini scores (r=-0.416, P<0.05), the level of MMP-9(r= - 0.503, P < 0.05) were correlated negatively with the level of PPARγ. Conclusions Complicating with T2DM can aggravate coronary artery disease and plaque instability degree in ACS patients, and PPARγpossibly make an protective effect.
ABSTRACT
The human tissue kallikrein(KLK)gene family consists of 15 highly conservative serine pro-teases,which is the largest uninterrupted cluster of protease genes in the human genome. Several members of the family are expected to be markers for tumor diagnosis and prognosis. Studies find that the expressions of KLK2-11 and KLK13-15 are abnormal,and the majority of KLKs have potential diagnostic and prognostic values.
ABSTRACT
OBJECTIVE@#To investigate the change of CD4- CD25 regulatory T cells (Tregs) of peripheral blood in allergic rhinitis (AR) patients.@*METHOD@#T lymphocytes of twenty AR patients and eight healthy subjects were separated and the flow cytometry was used to measure the percentage of CD4+ CD25+ Treg.@*RESULT@#Compared with control group, the percentage of Treg and CD4- CD25high Tregs of AR patients was decreased significantly (P < 0.05).@*CONCLUSION@#The proportion of CD4+ CD25+ Tregs decreased in AR patients conspicuously, which might be one of the pathogenesis of AR.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Case-Control Studies , Flow Cytometry , Interleukin-2 Receptor alpha Subunit , Allergy and Immunology , Metabolism , Rhinitis, Allergic, Perennial , Blood , Allergy and Immunology , T-Lymphocytes, Regulatory , Allergy and Immunology , MetabolismABSTRACT
Objective To investigate the significance of IL-18R? expression on CD4+ T cells in peripheral blood (PB) of children with Mycoplasma pneumoniae pneumonia (MPP).Methods T cell subtype CD3/CD4/CD8 and expression of IL-18R? on CD4+ T cells in PB from 35 children with MPP and 15 age- and sex-matched control subjects was determined by flow cytometry.Results Compared with that of healthy control, CD3+ and CD4+ positive cells in MPP children were decreased (P0.05).Conclusion There exists cell-mediated immune function disorders and Th1/Th2 imbalance in children with MPP. Th1 immune response is dominant in acute-stage MPP.
ABSTRACT
Objective To observe the clinical significance of detection of platelet-associated immunoglobulin (PAIgG).Methods Flow cytometry was used to detect PAIgG in 42 idiopathic throbocytopenic purpura (ITP) patients, 39 aplastic anemia (AA) patients, 21 patients with other autoimmune diseases and 20 normal controls. Rate of inefficient platelet infusion was calculated.Results Percentage of PAIgG positive in ITP, AA and other autoimmune diseases was higher than that in normal controls (P
ABSTRACT
Objective To develop a real-time fluorescence quantitative PCR(FQ-RCR)method for detection of Kallkreins(KLK)4 gene expression in human breast cancer,and to investigate KLK4 expression levels in breast cancer and normal tissues.Methods KLK4 expression levels of 25 normal breast tissues and 39 cancer tissues were detected by the developed real-time quantitative PCR method.The statistical analysis for the relationship between KLK4 expression and several pathological parameters was performed by t test.Results The levels of KLK4 mRNA in normal breast and breast cancer tissue were 0.0120?0.0044 and 0.0272?0.0067 respectively(P0.05).Conclusions The level of KLK4 mRNA in breast cancer tissue was higher significantly than that in normal breast tissue.The results indicated that KLK4 gene expression may have relevance with breast cancer development but have no significant relevance with ER,PR,CerbB-2 and tumor metastasis.
ABSTRACT
Expression levels of TGF-β1 mRNA in renal cortex of control group, diabetic group and taurine group measured by real-time quantitative RT-PCR were (7.0±0.8)×10 -3, (64.4±8.0)×10 -3 and (16.7±2.0)×10 -3, respectively. The corresponding results with end-point RT-PCR method were 0.28±0.12,0.58±0.16 and 0.43±0.10, respectively. Compared with end-point RT-PCR method, real-time quantitative with SYBR Green I was easier, faster, sensitive and specific.
ABSTRACT
Objective To investigate the relationship between herpes simplex virus(HSV) and spontaneous abortion. Methods 40 aborted material and 16 post-aborted curettings were tested for the presence of HSVDNA with in situ hybridization.In addition 20 normal endometria and 20 induced abortive placentas were used as controls. Results 56.3 %(9/16)post-abortion curettings were demonstrated the presence of HSVDNA.It was higher than that of the control group( P
ABSTRACT
objective To quantify the expression of kallikrein gene 6 (KLK6) in malignant and benign breast tissues and to statistically analyze the relationship between KLK6 expression levels and the clinico-pathological variables in patients with breast cancer. Method Paired breast tissue samples of cancerous and corresponding non-cancerous tissues were obtained from 48 patients with breast cancer who underwent surgical resection. Quantitative analysis of KLK6 mRNA expression was done using real-time quantitative reverse transcription-PCR (RT-PCR). The relationship between KLK6 expression intensity and various clinico-pathological variables was analyzed. In addition, the technique of small interfering RNA-mediated gene silencing was used to suppress the KLK6 expression in MCF-7, and to investigate the effects of KLK6 on cell proliferation rate and cell cycle. Results KLK6 mRNA over expression was found in cancerous tissues in 35 out of 48 patients (73%) as compared with normal breast tissue. The mean expression level of KLK6 mRNA in cancerous tissues was significantly higher than that in non-cancerous tissues (P
ABSTRACT
Objective To develop a real-time quantitative PCR method for detection of human breast cancer related novel gene-kallikrein gene 6 (klk6) expression and investigate klk6 expression levels in breast cancer tissue.Methods Using Sybr Green I, with GAPDH as reference, a real-time quantitative PCR method was established. klk6 expression levels of 32 normal and breast cancer tissues were detected and analyzed by the method and REST software.Results The amplification efficiencies for GAPDH and klk6 of the real-time quantitative PCR method were 0.90 And 0.95, respectively; inter-coefficient of variation were 1.0%~2.1% and 0.8%~1.2%,respectively; intra-coefficient of variation were 3.2% and 3.9%, respectively. The relative expression levels of klk6 in normal breast and breast cancer tissues were 0.017?0.009 and 0.040?0.017 with GAPDH as reference. Analysis results with REST indicated klk6 expression was up- regulated in breast cancer.Conclusion The real-time quantitative PCR method with Sybr Green I for klk6 mRNA quantification was simple, specific, reproductive and reliable, and could be used to study relationship betweeen klk6 expression and tumor.
ABSTRACT
Objective The purpose of the current study was to develop a real-time quantitative reverse transcription-PCR method for detection of human kallikrein gene 5(KLK5) mRNA expression,quantify the expression of KLK5 mRNA in malignant and benign breast tissues,and statistically analyze whether KLK5 expression levels correlate with clinicopathologic variables in the patients with breast cancer.Methods Paired breast tissue samples from cancerous and corresponding noncancerous tissues were obtained from 48 patients with breast cancer who underwent surgical resection.The relationship of KLK5 mRNA expression status with various clinicopathologic variables were quantitativly analyzed by real-time RT-PCR.Results KLK5 mRNA expression in cancerous tissues was decreased in 38 of 48(79%) breast cancer patients compared with normal counterparts.The mean expression level of KLK5 mRNA in cancerous tissues was significantly lower than that in noncancerous tissues(P0.05).Conclusion The results of this study indicated that KLK5 mRNA expression was significantly lower in cancerous tissues than that in noncancerous breast tissues,and low expression of KLK5 mRNA correlated with TMN stage,metastasis and ER status.