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Objective@#To investigate the relationship between miRNA-196b-5p and miRNA-99a-5p expression and autophagy and apoptosis in multiple myeloma cells.@*Methods@#Human myeloma cell line U266 and normal CD138+ plasma cells were selected as the research objects. The subjects were divided into 45 cases of multiple myeloma patients and 40 healthy controls. The expression of miRNA-196b-5p and miRNA-99a-5p was measured by real-time quantitative PCR, and Western blot was used to determine the expression of autophagy related protein LC3-Ⅱ, LC3-Ⅰ, P62, Beclin-1 expression, apoptosis related protein CL caspase3, CL caspase7, Bcl-2, Bax, and TGF-β/Smad pathway associated proteins TGF-β1, Smad2/3, p-Smad3 and Smad7. The cell apoptosis rate was determined by flow cytometry. The correlation between miRNA expression level and clinical characteristics of multiple myeloma patients was analyzed.@*Results@#Compared with normal plasma cells, the expression of miRNA-196b-5p in myeloma cells increased significantly (0.43±0.15 vs 2.44±0.63 or 2.02±0.85, all P<0.001), the expression of miRNA-99a-5p was significantly decreased (1.87±0.61 vs 0.62±0.15 or 0.80±0.33, P<0.001), LC3-Ⅱ/LC3-Ⅰ increased significantly (P<0.05), Beclin-1 expression increased significantly (P<0.05), P62 expression decreased significantly (P<0.05). The expression of Bax, CL caspase3 and CL caspase7 decreased significantly (P<0.05), and the expression of Bcl-2 increased significantly (P<0.05) and apoptosis rate significantly decreased (P<0.05). After transfected with miRNA-196b-5p mimic or miRNA-99a-5p inhibitor, the LC3-Ⅱ/LC3-Ⅰ of CD138+ plasma cells increased significantly (P<0.05), the expression of Beclin-1 increased significantly (P<0.05), P62 expression decreased significantly (P<0.05), and the apoptosis rate significantly decreased (P<0.05). However, after autophagy inhibitor of 3-MA was administered, the apoptotic rate of the above reaction system did not change significantly (P>0.05). The expression of miRNA-196b-5p and miRNA-99a-5p was significantly correlated with DS and ISS stage in multiple myeloma patients (P<0.05).@*Conclusion@#miRNA-196b-5p and miRNA-99a-5p are closely related to the clinical characteristics of patients with multiple myeloma. The overexpression of miRNA-196b-5p and down regulation of miRNA-99a-5p could inhibit the apoptosis of myeloma cells by up regulation of autophagy, and the mechanism is related to the activation of the TGF-β/Smad signaling pathway.
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AIM:To investigate the relationship between the expression of adducin 3 (ADD3) and its splicing isoforms and colorectal cancer (CRC).METHODS:The expression of ADD3, ADD3-Ia and ADD3-Ib in 50 pair of CRC tissues , 20 pairs of colorectal polyp tissues , and 2 CRC cell lines SW480 and SW620 before and after oxaliplatin or fluoroura-cil intervention were detected by real-time PCR.The cell activity was determined by MTT assay , the cell migration ability was evaluated by wound-healing assay , and the cell invasion ability was measured by Transwell assay .RESULTS:The expres-sion levels of ADD3 and ADD3-Ib were decreased in the CRC tissues as compared with the normal mucous (P<0.01), and ADD3-Ia/Ib ratio was increased in the CRC tissues (P<0.01).The expression level of ADD3-Ia was higher in T3-4 group than that in T1-2 group (P<0.05).Reduced expression of ADD3, ADD3-Ia and ADD3-Ib in colorectal polyps was observed compared with the normal tissues (P<0.01).Compared with the SW480 cells, the expression levels of ADD3-Ia and ADD3-Ib were lower (P<0.05) and the ADD3-Ia/Ib ratio was higher (P <0.01) in the SW620 cells.After treated with oxalipla-tin or fluorouracil, the cell activity, migration and invasion in the SW620 and SW480 cells were weakened accompanied by the increases in the expression levels of ADD 3, ADD3-Ia and ADD3-Ib to various certain extents .CONCLUSION:In CRC there is a tendency that ADD3-Ib reduction leads to ADD3 decrease, accompanied by an increased ADD3-Ia/Ib ratio.The expression changes of ADD 3 and its splicing isoforms in the CRC may be relevant to its invasion ability .
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eEF1A2 might be a putative oncoprotein in HCC .eEF1A2 over-expression has noticeable effects on the HCC cell prolifera-tion enhancement , differentiation inhibition , and cell cycle acceleration through the G 0/G1 phase to S phase and G 2/M phases.
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Objective To analyze the risk factors of 30-day mortality in patients with malignant hilar obstruction (MHO) after percutaneous transhepatic biliary metal stent deployment. Methods One hundred and fifty-nine consecutive patients with MHO caused by cholangiocarcinoma or gallbladder carcinoma were enrolled in this study. Percutaneous transhepatic biliary stent (PTBS) implantation was carried out in all the patients. Independent predictors for 30-day mortality were evaluated by logistic regression analysis. Covariates that were incorporated into the multivariate analysis were the variables that reached statistical significance (P < 0.1) in univariate analysis. Two-tailed, P value of less than 0.05 was considered to be statistically significant. Results The 30-day mortality of patient with MHO after metal stent deployment was 9.4%. Univariate analysis indicated that the differences in WBC (OR = 1.224.95%CI [1.07 - 1.44], P < 0.01), INR (OR=78.75, 95%CI [5.02-1 235.70], P<0.01), PT(OR=1.55, 95%CI [1.18-2.04], P<0.01), BUN (OR=1.19, 95%CI [1.02- 1.38], P < 0.05), CRE(OR = 1.02, 95%CI [1.000 - 1.041], P < 0.1) and lymph nodes metastasis(OR = 0.334. 95%CI[0.105 - 1.131], P < 0.1) were statistically significantly between 30-day mortality group and non-30-day mortality group. Multivariate analysis showed that statistically significant differences in WBC (OR = 1.19, 95%CI[1.026 - 1.380], P < 0.05), INR(OR = 151.5, 95%CI [3.13 - 5 440.7], P < 0.05) and CRE (OR = 1.025, 95%CI [1.002 - 1.048], P < 0.05) also existed palliative treatment for patients with malignant hilar obstruction. Active preoperative measures to improve hepatic and renal functions as well as to control infection are necessary in order to reduce 30-day mortality.
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Objective To establish real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) for human coronaviruses(HCoV)-NL63,HCoV-HKU1,HCoV-OC43 and HCoV-229E,and to investigate the prevalence of the four coronaviruses in children with acute respiratory tract infection(ARTI) in Fuzhou area.Methods Totally of 538 nasopharyngeal swabs were collected from pediatric patients with ARTI,including 289 specimens from children with acute upper respiratory tract infection (AURTI) and 249 from acute lower respiratory tract infection (AURTI) during three consecutive winter-spring seasons from December to April of 2006 - 2009 in Fuzhou area.All the specimens were subjected to FQ-PCR specific for HCoV-NL63,HCoV-HKU1,HCoV-OC43 and HCoV-229E,respectively.The enumeration data were analyzed by chi square test.Results The FQ-PCR methods were established for detecting HCoV-NL63,HCoV-HKU1,HCoV-OCA3 and HCoV-229E.The intraassay coefficient of variation (CV) and interassay CV were both ≤ 1.6％.The coronaviruses were detected in 41 (7.6％) children with ARTI,including HCoV-NL63 in 8 (1.5％)children (1 with AURTI,7 with ALRTI),HCoV-229E in 5 (0.9％; 1 with AURTI,4 with ALRTI),HCoV-HKU1 in 6 (1.1％; 1 with AURTI,5 with ALRTI),and HCoV-OC43 in 22(4.1％; 13 with AURTI,9 with ALRTI).The four coronaviruses were detected during each of the three winter-spring seasons and the positive rates of different periods were not significantly different (P＞0.05).The HCoV-OC43 positive rate was significantly higher than HCoV-NL63,HCoV-229E and HCoV-HKU1 (x2 =6.721,10.979,9.387; respectively; all P＜0.01).ConclusionsIt is suggested that the four coronaviruses might be important virus pathogens in children with ARTI in Fuzhou,China.And detection of them is needed for etiology and epidemiology evaluations for children with ARTI.
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Objective To study the effects of exercises training on the recovery of neurological function and the expression of matrix metalloproteinase-9 (MMP-9) in perihematomal brain tissue after intra cerebral hemorrhage (ICH) in rats. Methods Sixty-four male adult Sprague-Dawley rats were randomly divided into exercises group and control group. ICH model was induced by autobiood injection. The rats in exercises group were given balance, grasping and walking exercises every day. The rats in control group exercised freely in standard cages. Neurological function in both groups was measured at the 3rd, 7th, 14th and 21st d. All the rats were sacrificed and the concentration of MMP-9 was measured with immunohistochemical method and in situhybridization. Results In both groups neurological function scores was lowest at the 3rd d, were highest at the 21st d. There was no significant difference between two groups at the 3rd d, but at the 7th, 14th and 21st d the differences between two groups were significant ( P < 0.05 ). In exercises group, MMP-9 expressed weakly at the 3rd d, and peaked at the 7th d, then declined gradually. In control group, MMP-9 expression peaked at the 3rd d, then decreased slowly. MMP-9 protein expression was in accord with mRNA. MMP-9 expression in exer-cises group was significantly different from control group on the 3rd and 7th d (P < 0.05 ) , but not at the 14th and 21st d ( P > 0.05 ). Conclusions Early exercises can inhibit the expression of MMP-9 that could play a role in protecting neurons. Daily exercises can stimulates the expression of MMP-9 so as to have a positive role in midstage of disease. MMP-9 may be involved in tissue remodelling and vascular repairing, which prompt neu-rofunction recovery.
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Objective To clone and express human autoantigen Sm B'in methylotrophie yeast Pichia Pagtoris.Methods The gene Sm B' was cloned bv PCR The PCR product wag inserted into the vector pPIC9k.The recombinant plasmid pPIC9k.Sm B' was transformed into yeast Sm D1168 by electroporation.The positive clones were screened in MD plates.The high copy number transformants were rapidly selected by using G418 and were induced by methan01.Supematants after induction were analyzed by SDS-PAGE and western blot.Sera collected from thirty patients with SLE.thirty patients with mixed connective tissue disease(MCTD)and thirty healthy volunteers were detected by immunodot and immunoblot.Results The PCR product wag about 700 bD in size which Wag in accordance with predicted 657 bp.The pPIC9k-Sm B'showed the same seqencing result with GenBank's report and restriction enzyme analysis confirmed our prediction.The pPIC9k-Sm B' positive clone produced a 32 000 protein which had natural immunogenicitv of human autoantigen Sm B'by SDS-PAGE and western blot.The positive rate of immunodot and IBT were 46.7%(42/90)and 51.1%(46/90),respectively.The agreement between immunodot and IBT was very close(Kappa value=0.911 2,P＜0.01).Conclusion Successfully cloning and expression of human autoantigen Sm B' in methylotmphic yeast Pichia Pagtoris hid a foundation for further research work.
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Systemic lupus erythematosus(SLE) is an autoimmune disease characterized by the production of autoantibodies against a wide spectrum of nuclear, cytoplasmic,and cell membrane autoantigens.Among them,the anti-Smith antibodies are specific for SLE and thus have been included as a criteria for diagnosis.They are not only a diagnostic marker but also a reliable measure of disease activity in SLE,we intend to describe the biological functions and clinical significance of the Smith antigen and the detection of anti-Smith antibodies.