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Objective:To investigate the correlation between collateral circulation and infarct pattern and outcome in acute ischemic stroke patients with anterior circulation intracranial atherosclerosis.Methods:Acute ischemic stroke patients with anterior circulation intracranial atherosclerotic severe stenosis or occlusion admitted to the Department of Neurology, the First Affiliated Hospital of Xinxiang Medical College from September 2018 to March 2020 were included prospectively. According to diffusion-weighted imaging, the infarct patterns were divided into perforator pattern, territorial pattern, watershed pattern, and mixed pattern. At 90 d after onset, the modified Rankin Scale was used to evaluate the outcome. 0-2 was defined as good outcome, and >2 was defined as poor outcome. Multivariate logistic regression analysis was used to determine the independent influencing factors of clinical outcome. Results:A total of 89 patients were enrolled, 50 (56.2%) had good collateral circulation and 39 (43.8%) had poor collateral circulation. The distribution patterns of infarct: 22 patients (24.7%) were perforator pattern, 26 (29.2%) were territorial pattern, 17 (19.1%) were watershed pattern, and 24 (30.0%) were mixed pattern. The proportion of patients with good collateral circulation was 81.8%, 65.4%, 29.4% and 41.7%, respectively in the perforator pattern group, territorial pattern group, watershed pattern group, and mixed pattern group. Good collateral circulation was more common in the perforator pattern group, and poor collateral circulation was more common in the watershed pattern group. At 90 d after onset, 53 patients (59.6%) had a good outcome and 36 (40.4%) had a poor outcome. The baseline homocysteine level in the good outcome group was significantly lower than that in the poor outcome group (17.91±4.62 μmol/L vs. 20.35±4.67 μmol/L; t=2.436, P=0.017), and the proportion of patients with good collateral circulation was significantly higher than that of patients with poor outcome (73.6% vs. 30.6%; χ2=16.124, P<0.001). Multivariate logistic regression analysis showed that higher homocysteine level was an independent risk factor for poor outcome (odds ratio 1.174, 95% confidence interval 1.061-1.298; P=0.002) and good collateral circulation was an independent protective factor for good outcome (odds ratio 0.095, 95% confidence interval 0.038-0.239; P<0.001). Conclusions:Good collateral circulation was more common in patients with perforator pattern, and poor collateral circulation was more common in patients with watershed pattern. Good collateral circulation was independently associated with the good clinical outcome in acute ischemic stroke patients with anterior circulation intracranial atherosclerosis.
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Background@#Coinfection with avian leukosis virus subgroup J (ALV-J) and reticuloendotheliosis virus (REV) is common in chickens, and the molecular mechanism of the synergistic pathogenic effects of the coinfection is not clear. Exosomes have been identified as new players in the pathogenesis of retroviruses. The different functions of exosomes depend on their cargo components. @*Objectives@#The aim of this study was to investigate the function of co-regulation differentially expressed proteins in exosomes on coinfection of ALV-J and REV. @*Methods@#Here, viral replication in CEF cells infected with ALV-J, REV or both was detected by immunofluorescence microscopy. Then, we analyzed the exosomes isolated from supernatants of chicken embryo fibroblast (CEF) cells single infected and coinfected with ALV-J and REV by mass spectrometry. KEGG pathway enrichment analyzed the co-regulation differentially expressed proteins in exosomes. Next, we silenced and overexpressed tripartite motif containing 62 (TRIM62) to evaluate the effects of TRIM62 on viral replication and the expression levels of NCK-association proteins 1 (NCKAP1) and actin-related 2/3 complex subunit 5 (ARPC5) determined by quantitative reverse transcription polymerase chain reaction. @*Results@#The results showed that coinfection of ALV-J and REV promoted the replication of each other. Thirty proteins, including TRIM62, NCK-association proteins 1 (NCKAP1, also known as Nap125), and Arp2/3-5, ARPC5, were identified. NCKAP1 and ARPC5 were involved in the actin cytoskeleton pathway. TRIM62 negatively regulated viral replication and that the inhibition of REV was more significant than that on ALV-J in CEF cells coinfected with TRIM62. In addition, TRIM62 decreased the expression of NCKAP1 and increased the expression of ARPC5 in coinfected CEF cells. @*Conclusions@#Collectively, our results indicated that coinfection with ALV-J and REV competitively promoted each other's replication, the actin cytoskeleton played an important role in the coinfection mechanism, and TRIM62 regulated the actin cytoskeleton.
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At present, nucleic acid testing technology has been widely used in clinical laboratory diagnosis. Conventional detection technique such as real-time PCR is complicated, time consuming, and dependent on specific instruments. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas) system is an adaptive immune defense system against viruses in bacteria and archaea, which has been developed into a powerful technology for genome editing. Recently, the leading groups engaged in CRISPR have set up new tools for nucleic acid detection based on Cas13a, Cas12a and newly discovered protein-Cas14, which plays an important role in rapid diagnosis of infectious diseases, detection of gene mutations in cancer and genotyping. Since they are ultrasensitive, specific, rapid and cost-effective, it is expected to bring great potential for molecular diagnosis. In this review, the mechanism of CRISPR/Cas system and the principle, the applications of the newly-developed diagnostic platforms are introduced. What′s more, the advantages, limitations and prospects of the technologies are summarized.
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Objective:To observe the effect of irradiation on the production of IL-8 in lung cancer cell line A549 and explore its possible mechanism.Methods:A549 cells irradiated with different doses of X-rays were used to collect cell supernatant, cellular RNA and protein at different time points after irradiation. The expression level of IL-8 mRNA in A549 cells after irradiation was detected by RT-PCR, which was further validated by real-time quantitative PCR. The expression level of IL-8 in the cell supernatant was quantitatively measured by ELISA. The expression levels of cellular signaling pathway molecules in A549 cells after irradiation were detected by Western Blot. The A549 cells were pretreated with p38 MAPK inhibitor, NF-κB inhibitor and ROS scavenger. The effect of these inhibitors on the expression of IL-8 in A549 cells induced by irradiation was evaluated by ELISA.Results:Irradiation up-regulated the expression of IL-8 in A549 cells in a dose-and time-dependent manner. Irradiation activated the p38 MAPK and NF-κB signaling pathway in A549 cells. p38 MAPK and NF-κB inhibitors blocked the induction of IL-8 of A549 cells by irradiation. Inhibition of ROS failed to inhibit the induction of IL-8 of A549 cells by irradiation.Conclusion:Irradiation can increase the production of IL-8 in lung cancer cells A549, possibly through the activation of p38 MAPK and NF-κB signaling pathways in a ROS-independent pattern.
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At present, nucleic acid testing technology has been widely used in clinical laboratory diagnosis. Conventional detection technique such as real-time PCR is complicated, time consuming, and dependent on specific instruments. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas) system is an adaptive immune defense system against viruses in bacteria and archaea, which has been developed into a powerful technology for genome editing. Recently, the leading groups engaged in CRISPR have set up new tools for nucleic acid detection based on Cas13a, Cas12a and newly discovered protein-Cas14, which plays an important role in rapid diagnosis of infectious diseases, detection of gene mutations in cancer and genotyping. Since they are ultrasensitive, specific, rapid and cost-effective, it is expected to bring great potential for molecular diagnosis. In this review, the mechanism of CRISPR/Cas system and the principle, the applications of the newly-developed diagnostic platforms are introduced. What′s more, the advantages, limitations and prospects of the technologies are summarized.
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Non-coding RNAs (ncRNAs) are genomic transcripts that do not encode proteins.They are also closely associated with tumor initiation and progression,as they play regulatory roles in gene transcription,post-transcription,and translation.Moreover,ncRNAs may enter circulatory system in the form of microvesicles or exosomes,or in combination with protein.The circulating ncRNAs are stable and widely existed in body fluids such as blood and urine.Certain circulating ncRNAs are differentially expressed in tumors,suggesting that they have potential value as novel tumor biomarkers.This article briefly reviews the progress of circulating microRNA (miRNA),long non-coding RNA (lncRNA) and circular RNA (circRNA) as breast cancer biomarkers,and discusses the clinical value of circulating ncRNA as a novel biomarker for breast cancer diagnosis and prognosis.
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Objective To analyze the identification, drug resistance and clinical significance of a rare bacterium of Bordetella holmesii ( B. holmesii) to improve its detection and clinical diagnosis and treat-ment. Methods A strain isolated from a bacteremia case was identified by bacterial culture, biochemical tests and 16S rRNA gene sequencing. Mega 7. 0 software was used to conduct a similarity analysis of 16S rRNA gene sequences between the type strains of Bordetella spp. and the isolate, and then a phylogenetic tree was constructed. Antibiotic resistance of the isolate was determined by E-test. Changes in bacterial growth were measured after adding different concentrations of riboflavin or its inhibitor lumiflavin to the cul-ture medium. Results B. holmesii ABD2 was the pathogen causing bacteremia in the immunocompetent pa-tient. It was deposited under the number of CGMCC 1. 13721 in China General Microbiological Culture Col-lection Center (CGMCC), and the 16S rRNA gene sequences were deposited in National Center for Biotech-nology Information ( NCBI) with the accession number of KT828544. 1. Unrooted tree showed that the B. holmesii strain was highly homologous with B. pertussis. Antibiotic susceptibility test showed that the mini-mum inhibitory concentrations ( MIC) of piperacillin, ceftazidime, cefepime, imipenem, meropenem, cipro-floxacin, levofloxacin, gentamicin, amikacin, erythromycin, tetracycline and polymyxin B against the isolate were low, while the MIC values of cefazolin, cefuroxime, cefoxitin, cefotaxime, aztreonam and trime-thoprim-sulfamethoxazole were high. Riboflavin accelerated the growth of B. holmesii ABD2, while its inhibi-tor lumiflavin had an inhibitory effect. Conclusions As B. holmesii is hard to isolate and identify, limited clinical, microbiological and epidemiological data are available. It is an under-recognized pathogen with a considerable amount of information that remains to be studied.
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Phosphatidylinositol 3-kinase (PI3K) signaling plays an important role in the regulation of cellular lipid metabolism and non-alcoholic fatty liver disease (NAFLD). However, little is known about the role of the regulatory subunits of PI3K in lipid metabolism and NAFLD. In this study, we characterized the functional role of PIK3R3 in fasting-induced hepatic lipid metabolism. In this study, we showed that the overexpression of PIK3R3 promoted hepatic fatty acid oxidation via PIK3R3-induced expression of PPARα, thus improving the fatty liver phenotype in high-fat diet (HFD)-induced mice. By contrast, hepatic PIK3R3 knockout in normal mice led to increased hepatic TG levels. Our study also showed that PIK3R3-induced expression of PPARα was dependent on HNF4α. The novel PIK3R3-HNF4α-PPARα signaling axis plays a significant role in hepatic lipid metabolism. As the activation of PIK3R3 decreased hepatosteatosis, PIK3R3 can be considered a promising novel target for developing NAFLD and metabolic syndrome therapies.
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Animals , Mice , Diet, High-Fat , Fatty Liver , Lipid Metabolism , Non-alcoholic Fatty Liver Disease , Phenotype , Phosphatidylinositol 3-KinaseABSTRACT
Objective To study the clinical effect of montelukast combined with sidenafil in the treatment of chronic obstructive pulmonary disease ( COPD ) complicated with pulmonary hypertension .Methods 94 COPD patients with pulmonary hypertension were selected ,and they were randomly divided into two groups according to the digital table,47 cases in each group .The conventional group was treated with conventional treatment ,the combination group was treated with montelukast sodium and sildenafil on the basis of the conventional treatment .The effect,safety, quality of life(SF-36),heart and lung function were compared in two groups .Results The total effective rate of the combination group was 95.74％,which was significantly higher than 78.72％of the conventional group ,the difference was statistically significant (χ2 =6.114,P <0.05).Compared with the conventional group,6 minutes walking distance ,mean pulmonary artery pressure , forced expiratory volume in one second of the combination group were significantly improved,the differences were statistically significant (t =6.244,7.182,7.034,all P <0.05).After treatment,the SF-36 score of the combination group was (84.78 ±13.61)points,which was significantly higher than (63.47 ±9.62)points of the conventional group (t=9.343,P<0.05).The incidence rate of adverse reaction had no statistically significant difference between the two groups (χ2 =0.000,P >0.05).Conclusion Montelukast and sildenafil in the treatment of COPD patients complicated with pulmonary hypertension has good clinical effect ,and it can effectively improve patients'cardiopulmonary function , exercise endurance and quality of life , has no significant side effects.
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Objective To study the risk of bloodstream infection in relation with peripherally inserted central venous catheter (PICC) in hospitalized non-cancer patients.Methods Clinical data of 172 non-cancer patients admitted to our hospital for PICC were collected.The risk of PICC-related bloodstream infection (CRBSI) in domestic hospitalized non-cancer patients was analyzed and systematically assessed.Results Of the 183 PICCs placed in 172 patients included in this study,61.7％ were placed in general wards,38.3％ were placed in ICU,87.8％ were placed in combination with indwelling urinary catheter,29.7％ were placed in combination with mechanical ventilation.The median PICC indwelling time was 35 days.CRBSI occurred in 6 patients with an incidence of 0.6/1000 PICCs/day.The risk of CRBSI was centralized in domestic tumor patients after PICC.The reported CRBSI was significantly different in hospitalized non-cancer patients (0.26-33.10/100 PICC).Conclusion The risk of CRBSI is lower in hospitalized patients after PICC placement than after traditional central venous catheter placement.Further studies are needed to assess its value in ICU.
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Objective To investigate the potential role of CXC chemokine ligand 16 (CXCL16)/CXC chemokine receptor 6 (CXCR6) pathway in the progression of diabetic nephropathy (DN). Methods 8?week old male db/db mice were randomly divided into DN group and DN inflamed group. 10% casein was subcutaneously injected to induce the DN mouse model with inflammation. In vitro, HK?2 cells were treated with high glucose (HG), and IL?1β+HG to investigate the effect of inflammatory stress on HK?2 cells. Further knockdown CXCL16 was mediated by RNA interference to determine the effects of CXCl16, then cells were divided into HG+IL?1βgroup, HG+IL?1β + siCXCL16 group and HG + IL?1β + vehicle group. Changes of renal function in mice were assessed by 24 h proteinuria and N?acetyl?β?D?glucosaminidase (NAG) during 8 weeks. The ultra?microstructure was checked by electron microscopy at 8th week. Lipid accumulation in kidneys and HK?2 were observed by Filipin staining and quantitative assay of intracellular free cholesterol. The protein expressions of CXCl16, CXCR6, a disintegrin and metalloproteinase?10 (ADAM10), fibronectin and α smooth muscle actin (α?SMA) in renal tissue were detected by immunohistochemistry and Western blotting. The mRNA and protein expressions of CXCl16, CXCR6, ADAM10, fibronectin andα?SMA in HK?2 cells were detected by real?time PCR and Western blotting, and protein expressions of CXCl16, CXCR6 and ADAM10 in HK?2 cells were also tested by cell immunofluorescence. Results Mice in DN inflamed group had higher 24 h proteinuria and NAG than those in DN group, and the differences between two groups shown statistical significance at 8th week (all P<0.05). Compared with DN mice, DN inflamed mice had more vacuoles within renal tubular cells, with mitochondrial swelling, deformation and decrease. Lipid accumulation and protein expressions of fibronectin and α?SMA were increased in DN inflamed group when compared with DN group (all P<0.05). Further, the expressions of CXCL16, CXCR6, ADAM10 were significantly increased in DN inflamed group (all P<0.05). In vitro, the mRNA and protein expressions of CXCL16, CXCR6, ADAM10, fibronectin and α?SMA, and lipid accumulation were increased in high glucose plus IL?1βgroup when compared with high glucose group (all P<0.05). However, after siRNA of CXCL16 transfection, the mRNA and protein expressions of CXCL16, CXCR6, ADAM10, fibronectin andα?SMA were down?regulated in HG+IL?1β+siCXCL16 group as compared with high glucose+IL?1βgroup (all P<0.05). Furthermore, lipid accumulation was decreased (P<0.05). Conclusion Inflammation accelerates tubulointerstitial injury in DN partly through the activation of CXCL16 pathway, which may facilitate the lipid accumulation in tubular epithelial cells.
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BACKGROUND: The mechanisms of spinal cord ischemia-reperfusion injury are the result of the combined effects of multiple factors, but there is no effective treatment. OBJECTIVE: To investigate the effect of lipoxin receptor agonist BML-111 on inflammatory factor and apoptosis in rats with spinal cord ischemia-reperfusion injury. METHODS: A total of 72 healthy adult male Sprague-Dawley rats were randomly divided into sham surgery group, ischemia-reperfusion group and BML-111 group. Rat models of spinal cord ischemia-reperfusion injury were established by clamping the abdominal aorta in the later two groups. Rats in the ischemia-reperfusion group and BML-111 group were injected with 0.1 mL of saline and 1 mg/kg BML-111 through caudal vein at 30 minutes after model establishment. RESULTS AND CONCLUSION: Compared with ischemia-reperfusion group, BBB scores were significantly improved, pathological injury of spinal cord tissue significantly reduced, the number of apoptotic cel s, tumor necrosis factor α and interleukin-1β expression, myeloperoxidase oxide activity and malondialdehyde content decreased in the BML-111 group. These findings indicate that lipoxin receptor agonist BML-111 can inhibit neuronal apoptosis and inflammation so as to reduce spinal cord injury.
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Objective To explore the possibility of rapamycin to up-regulate radiosensitivity of nasopharyngeal carcinoma (NPC) and its molecular mechanism.Methods In vitro,with untreated cells as the control,NPC cells were treated with rapamycin,irradiation (IR),or both rapamycin and IR.Phosphorylation of S6 and GSK3β,expression of Cyclin D1,clonogenic survival,number of residual γH2AX foci,and cell cycle status between study groups were compared.In vivo,athymic mice bearing CNE1 tumor were similarly treated.Tumor weight,Cyclin D1 and phosphorylated S6 in the xenograft model were compared between study groups.Results The results showed that rapamycin alone decreased the phosphorylation of S6 and glycogen synthase kinase 3 β (GSK3β),and the expression of Cyclin D1 in NPC cells.Thus,rapamycin-treated NPC cells had lower cell viability,higher DNA damage and more G1 arrest than the control,which was reflected by the in vivo study that rapamycin significantly attenuated tumor growth and decreased the levels of Cyclin D1 and phosphorylated S6.Moreover,the combination of rapamycin and IR caused the highest cell death,DNA damage,G1 arrest and tumor regression compared to those treated either alone.Conclusions Rapamycin up-regulate NPC radiosensitivity by inhibiting signal transduction of Akt/mammalian target of rapamycin (mTOR)/S6 pathway and Akt/GSK3β pathway,and by downregulating Cyclin D1 expression.
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Avian leukosis virus subgroup J (ALV-J) is an avian retrovirus that can induce myelocytomas. A high-frequency mutation in gene envelope endows ALV-J with the potential for cross-species transmission. We wished to ascertain if the ALV-J can spread across species under selection pressure in susceptible and resistant hosts. First, we inoculated (in turn) two susceptible host birds (specific pathogen-free (SPF) chickens and turkeys). Then, we inoculated three resistant hosts (pheasants, quails and ducks) to detect the viral shedding, pathologic changes, and genetic evolution of different isolates. We found that pheasants and quails were infected under the selective pressure that accumulates stepwise in different hosts, and that ducks were not infected. Infection rates for SPF chickens and turkeys were 100% (16/16), whereas those for pheasants and quails were 37.5% (6/16) and 11.1% (3/27). Infected hosts showed immune tolerance, and inflammation and tissue damage could be seen in the liver, spleen, kidneys and cardiovascular system. Non-synonymous mutation and synonymous ratio (NS/S) analyses revealed the NS/S in hypervariable region (hr) 2 of pheasants and quails was 2.5. That finding suggested that mutation of isolates in pheasants and quails was induced by selective pressure from the resistant host, and that the hr2 region is a critical domain in cross-species transmission of ALV-J. Sequencing showed that ALV-J isolates from turkeys, pheasants and quails had moved away from the original virus, and were closer to the ALV-J prototype strain HPRS-103. However, the HPRS-103 strain cannot infect pheasants and quails, so further studies are needed.
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Animals , Amino Acid Sequence , Avian Leukosis , Virology , Avian Leukosis Virus , Classification , Genetics , Physiology , Chickens , Ducks , Virology , Galliformes , Virology , Host Specificity , Molecular Sequence Data , Poultry Diseases , Virology , Quail , Virology , Sequence Alignment , Turkeys , Virology , Viral Envelope Proteins , Chemistry , Genetics , MetabolismABSTRACT
Objective To explore the function of p53 on regulating the expression of miR-148b in lung cancer cell line PC-9 and its corresponding molecular mechanism and the impact on cell proliferation. Methods Transient transfection of p53 eukaryotic expressing plasmids into lung cancer cell line PC-9 was performed to establish a cell model over-expressing p53. RT-PCR was used to explicit the impact of p53 on the expression of miR-148b. A reporter vector containing miR-148b promoter was used to investigate the function of p53 on regulating the transcription of miR-148b. Low-expressing miR-148b by transfecting its specific inhibitors , a CCK-8 assay was performed to explore the influence of miR-148b on the lung cancer cell proliferation inhibited by p53. Results Over-expression of p53 promoted miR-148b expression in lung cancer cell line PC-9. P53 could increase the luciferase activity driven by miR-148b promoters. Knockdown of miR-148b attenuated the impact of p53 on inhibiting the proliferation of PC-9 cells. Conclusion P53 inhibits the proliferation of lung cancer cell line PC-9 partially depending on miR-148b.
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The study was aimed to assess the effect of Klotho gene and sinoatrial node pacing channel gene (HCN4 and HCN2) for studying sick sinus syndrome, with Klotho gene under the interference of Plasmid-mediated short hairpin RNA. Twenty-five C57BL/6J mice were divided into four groups, i. e, plasmid shRNA 24h group, plasmid shRNA 12h group, sodium chloride 24h group and sodium chloride 12h group. Plasmid shRNA 50microL (1microg/microL) and sodium chloride 50microl were respectively injected according to mice vena caudalis into those in plasmid shRNA group and sodium chloride group. After 12h or 24h respectively, all mice were executed and their sinoatrial node tissues were cut. The mRNA of Klotho, HCN4 and HCN2 gene were detected by RT-PCR. The results of RT-PCR showed that Klotho, HCN4 and HCN2 mRNA levels were lower compared with those in sodium chloride 12h group after 12h interference interval. The results indicated that there might be the a certain relationship between Klotho gene and sinoatrial node pacing channel gene.
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Animals , Male , Mice , Glucuronidase , Genetics , Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels , Genetics , Metabolism , Mice, Inbred C57BL , Plasmids , Genetics , Potassium Channels , Genetics , Metabolism , RNA Interference , RNA, Messenger , Genetics , Metabolism , RNA, Small Interfering , Genetics , Sinoatrial Node , Metabolism , PhysiologyABSTRACT
Objective To detect the Bag-1 and VEGF expressions in HCC tissues,cancerous liver tissues and normal liver tissues,and to explore their relationship with prognosis.Methods Immunohistochemical staining (SP method) was used to detect Bag-1 and VEGF in 60 cases of hepatocellular carcinoma specimens,next to the 30 cases of hepatocellular carcinoma in liver tissue,and 14 cases of normal liver tissue specimens,and their clinicopathological features were analyzed.Results Bag-l,VEGF expression rates were highest in HCC tissues [80.0 ％ (48/60),73.3 ％ (44/60)] and with adjacent tissues [46.7 ％ (14/30),43.3 ％ (13/30)],normal liver tissue [28.6 ％ (4/14),21.4 ％ (3/14)] expression rates had differences (P ＜ 0.05).Bag-1,VEGF expression had relationship with liver cancer staging,lymph node metastasis,tumor thrombosis formation,the correlation between the presence or absence of capsule formation (P ＜ 0.05),while had no relationship with the patient gender,age,tumor grade,tumor size,and alpha-fetoprotein (AFP) and other factors correlation (P ＞ 0.05).Bag-1 and VEGF expression in HCC specimens positive 40 cases,while negative eight cases,both in HCC tissues expression was positively correlated (P ＜ 0.05).Bag-1,VEGF positive expression in patients 1-year,2-year survival rates were less than negative patients,the survival time of patients with positive group was significantly lower than the negative patients (P ＜ 0.05).Conclusion Bag-1 and VEGF expressions are closely related with biological characteristics of liver cancer,and which is closely related to the prognosis of patients with hepatocellular.
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Objective To prove the remarkable inhibitive effect of multiple siRNAs targeting a proliferation-inducing ligand (APRIL) on the human colorectal cancer cell.Methods We constructed a multiple short hairpin RNA(shRNA) expression vector containing four shRNAs (pG4) as well as four single one (pGsh644,pGsh1451,pGsh1938,pGsh2231) against APRIL gene in SW480 cell,and then transfected them into the human colorectal cancer cell line by cationic liposome.Ultimately,SW480 were screened by EGFP to obtain expression cell lines.APRIL expression levels including mRNA and APRIL protein were detected after transfected with all different kinds of vectors.Results A multiple shRNA expression vector containing four shRNAs (pG4) and four single ones were successfully constructed.Four single vectors (pGsh644,pGsh1451,pGsh1938,pGsh2231) and the multiple siRNAs expression vector (pG4) all decreased the APRIL mRNA by 56.2％,49.5％ ;50.9％,49.2％ and 79.3％.And APRIL protein expression was also remarkably reduced,especially by multiple siRNAs expression vector(87.5％).Conclusion Multiple siRNAs expression vector produced a more significant knockdown effect of APRIL than the vectors containing only one APRIL shRNA.What we found suggested us using the vector containing multiple shRNA to silence the expression of APRIL might be exploited as a novel therapeutic strategy for tumors.
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Objective To investigate the effects of a proliferation-inducing ligand(APRIL) on migration and invasion of colorectal cancer (CRC) and matrix metalloproteinases (MMPs) expression in order to observe the role of APRIL in CRC metastasis.Methods The siRNA plasmid vector targeting APRIL gene (siRNA-APRIL) was transfected into SW480 cells and recombinant human APRIL(rhAPRIL) was used to stimulate HCT-116 cells.Tumor cell migration and invasion were measured by Transwell chambers.RT-PCR and ELISA were applied to examine the expression level of MMPs.Results Metastatic and invasive capacities of siRNA-APRIL transfected SW480 were significantly inhibited,and these capacities of APRIL stimulated HCT-116 cells were significantly enhanced compared with their respective controls( all P＜0.05 ),accompanied with the alterations of MMPs mRNA and secreted protein expression( P＜0.05).The number of invading cells of SW480 control and rhAPRIL stimulated HCT-116 was significantly decreased by a MMP inhibitor GM6001 ( P＜0.05 ).Conclusion APRIL facilitates migration and invasion of CRC via regulation of MMPs,which suggests that APRIL might be used as a new target for the intervention and treatment of CRC metastasis.
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Objective To investigate the existence and variance of quinolone-resistance genes in a group of pan-drug resistant of Acinetobacter baumannii ( A.baumannii ).Methods Twenty strains of pandrug resistant A.baumannii were isolated from patients registered in Affiliated Hospital of Nantong University from January 2011 to April 2011.Drug target genes to quinolone (gyrA,parC) and quinolone-resistance genes mediated by mobile genetic elements [ qnrA,qnrB,qnrS,aac(6')-Ⅰ b-cr,qepA] were analyzed by PCR and verified by DNA sequencing.Results In all 20 strains of A.baumannii,the sense mutation was found in the quinolone resistance-determining region of the gyrA gene in the form of TCA to TTA at codon 83 (Ser-83-Leu).Moreover,in the quinolone resistance-determining region of the parC gene sense mutation was found in the form of TCG to TTG at codon 80 (Ser-80-Leu) and 3 synonymous mutations were CCC to CCT at codon 40,GTA to GT]T at codon 41 and CGT to CGC at codon 44.And parC gene was a new mutation.However,mutations were not found in quinolone-resistance genes mediated by mobile genetic elements [ qnrA,qnrB,qnrS,aac( 6 ' )-Ⅰ b-cr,qepA ].Conclusion Quinolone-resistance-determining region play a key role in resistance to quinolones in this group of A.baumannii.To our knowledge,this is first report about the emergence of the new mutation of parC gene in China.