Analysis of the expression of coxsackievirus and adenovirus receptor in five colon cancer cell lines

AIM： To investigate the expression of coxsackievirus and adenovirus receptor （CAR） and adenovirus-mediated reporter gene transfer in five human colon cancer cell lines.METHODS： Expression of CAR-specific mRNA and protein was analyzed by reverse transcriptase polymerase chain reaction and Western blotting, respectively. Adenovirusbased gene delivery was evaluated by infection of cells with adenoviral vector carrying the green fluorescent protein （GFP） gene.RESULTS： All the colon cancer cell lines examined （HT29, LS180, SW480, SW948 and SWlll6） expressed CAR full-length mRNA and an alternatively-spliced variant that lacks the transmembrane coding exon. All cell lines were detected as CAR-positive by Western blot analysis. Further, all cells we examined were efficiently infected with adenoviral vector-GFP.CONCLUSION： The data indicated that the five colon cancer cell lines tested expressed adenovirus primary receptor and could be efficiently infected by adenoviral vectors. Therefore, these cell lines will be useful for adenovirus-based gene transfer and research.

Expression of Coxsackievirus and Adenovirus Receptor in Human Lung Cancer： Possible Clinical Significance

OBJECTIVE To explore the relationship between CAR and the development of human lung cancer, as well as to provide the basis for the clinical treatment of lung cancer using an adenovirus vector-based gene therapy. METHODS CAR expression was assessed immunohisto- chemically in tumoral, paraneoplastic and normal samples from 112 lung cancer patients. At the same time, the mRNA and protein expression of CAR in 32 cases were determined by RT-PCR and Western blot. The relationship between CAR expression and clinicopathologic parameters was statistically analyzed. RESULTS There was no expression of CAR in normal lung tissue but a little in paraneoplastic tissue. The positive rate was 43% in squamous cell carcinoma, and 70% in adenocarcinoma. Both were much significantly higher than that in paraneoplastic tissue. The CAR expression level in adenocarcinoma was higher than that in squamous cell cancer, mRNA expression by RT-PCR and protein expression by Western blot were consistent with immunohistochemistry results. CONCLUSION CAR is overexpressed in human lung cancer, especially in adenocarcinoma. This data offer the reliable basis for adenovirus-mediated gene therapy of lung cancer; more important, CAR may take part in the formation or development of lung cancer; this may be exploitable for the development of antibody-directed therapy in human lung cancer.

Inhibitory Effect of Coxsackie Adenovirus Receptor on Invasion and Metastasis Phenotype of Ovarian Cancer Cell Line SKOV3

Full-length coxsackie adenovirus receptor (CAR) eukaryotic expression plasmid was transfected into an ovarian cell line, SKOV3, and its effect on the change of malignant metastasis phenotype was explored. CAR mRNA and protein expression levels among 4 ovarian cancer cell lines (A2780, SKOV3, SW626, CAOV3) and the positive control 293 (a transformed human embryo kidney cell line) was detected by using semi-quantitative RT-RCR and Western blot and compared. CAR-negative SKOV3 was transfected with the eukaryotic expression plasmid containing a full-length CAR cDNA and mock-vector respectively. The positive clones were screened by G418. The biological behavior changes of positive transfected cells were gauged by colony formation in soft agar assay and cell adhesion assay. Among the cell lines, there were obviously different CAR expression levels. CAR could not be detected in SKOV3. In transfected cell group, CAR expression was enhanced obviously as compared with non-transfected or mock-transfected groups. Cell adhesion in the transfected group was promoted. The number of colony formation was reduced significantly in transfected groups (25.32±8.91) as compared with that in non-transfected group (88.75±13.98) and mock-transfected group (82.53±19.37). Among the 4 ovarian cancer cell lines, CAR expression level was variable. Exogenous CAR expression had a potential role in inhibiting the malignant metastasis phenotype of ovary cancer cells.

Expression of coxsackie and adenovirus receptor in small cell lung cancer

Objective: We explored the expression of coxsackie and adenovirus receptor (CAR) in small cell lung cancer (SCLC) tissue. Methods: CAR expression in 31 SCLC was assessed in formaldehyde-fixed, paraffin-embedded tissue according to the EnVision immunohistochemistry procedure, while 3 samples of surgical specimens of non-malignant lung disease were taken as the negative control. Results: We observed that the expression of CAR was detectable positive in all the 31 cases from the small cell lung cancer tissue, in contrasting that non-malignant lung tissue control. Conclusion: The high expression of CAR appeared in SCLC tissue indicates that it play an important role in of adenovirus vector-based gene therapy in SCLC.

Expression and Significance of Coxsackie and Adenovirus Receptor in Renal-Cell Carcinoma

OBJECTIVE To investigate the expression of Coxsackie and Adenovirus receptor （CAR） in renal-cell carcinoma and the relationship of the CAR to the biological behavior of the carcinomas. METHODS The immunohistochemical SP method was used to detect the expression of Coxsackie and Adenovirus receptor in 48 cases of renal- cell carcinoma and in 12 cases of normal renal tissue 2 cm away from the tumor tissue. RESULTS The positive rates of CAR were 100% in 12 cases of para-tumor normal renal tissue and 35.4% in 48 cases of renal-cell carcinoma respectively. The difference of CAR expression between them was significant （P〈0.05）. The grades of the tumor were as follows： 22 in Grade Ⅰ, 17 in Grade Ⅱ and 9 in Grade Ⅲ with the CAR positive rate being 54.5%, 23.5% and 11.1%, respectively. There was a negative correlation between CAR expression and tumor grading （P〈0.05）. In addition, the number of the cases in stages I to IV were 19, 13, 11 and 5 respectively, with the respective positive rates being 57.9%, 30.8%, 18.2% and 0.0%, i.e. there also was a negative relationship between CAR expression and the stage （P〈0.05）. CONCLUSION CAR expression is down-regulated in renal-cell carcinoma compared with normal tissue. The level of CAR may be a sensitive predictor of differentiation, invasion and metastasis. Loss of CAR expression correlates with the invasive phenotype in our analysis of renal-cell carcinoma.

ROLE OF COXSACKIEVIRUS AND ADENOVIRUS RECEPTOR (CAR)IN CARDIOTOXICITY INFECTED BY COXSACKIEVIRUS B3

Objective To explore the role of coxsackievirus and adenovirus receptor（CAR） in cardiotoxicity infected by coxsackieviras B3. Methods A toxic cellular model was established in vitro by adding myocarditic coxsackievirus B3 （CVB3m） into the culture of neonatal mouse cardiomyocytes. 48 h later, the cardiomyocytes were divided into control, CVB3m, and CAR antibody ＋ CVB3m groups. CVB3m-mediated myocytopathic effect of above three groups was observed after further culturing for 48h. At the same time, the cardiomyocytes＇ viability of above three groups was assessed by MTT assay. Results The degree of cytopathic effect（CPE） of CAR antibody ＋ CVB3m group was significantly lower than CVB3m group （ P 〈 0. 01 ） and there was a significant increase in cell viability in CAR antibody ＋ CVB3m group compared with CVB3m group（ P 〈 0. 01 ）. No significant difference was found between CAR antibody ＋ CVB3m group and control group. Conclusion CAR antibody possesses a protective effect on CVB3m infected cardiomyoctyes, which indicates that CAR may play an important role in mediating cardiotoxicity infected by CVB3m.

Expression of Coxsackie and Adenovirurus Receptor and its Significance in Human Lung Cancer

OBJECTIVE To study the relationship between the coxsackie and adenovirus receptor （CAR） and the development of human lung cancer. To optimize adenovirus vector-based gene therapy.METHODS The expression of CAR in 112 cases of lung cancer was examined using immunohistochemistry. At the same time, the relationship between CAR expression and clinicopathologic characteristics was analyzed,RESULTS ：lhere is a little expression of CAR in normal lung tissue. Compared with paraneoplastic epithelial tissue of the lung, the expression of CAR is generally up-regulated in tumor tissues showing a significant dif- ference （P〈0.01）. The positive rate of CAR expression in squamous cell carcinoma was 43.1%, and in adenocarcinoma 70.2%, with the difference between the two rates being statistically significant （P〈0.01）. Compared to the paraneoplastic tissues, the difference in CAR positive expression was 35.4% for squamous cell carcinoma and 38.3% for adenocarcinoma. But the difference in different stages of squamous cell carcinoma had no statistical significance （P〉0.05）. However, the expression of CAR was at a high level in the bronchioalveolar carcinomas as 80.4% were CAR positive. This research showed that there was a specially high expression of CAR in adenocarcinomas.CONCLUSION CAR is expressed in human lungs at a low level and up-regulated in the tumor tissues, suggesting that there is a relationship between adenocarcinoma and CAR. This research provides a basis for planning a regimen of gene therapy using an adenovirus vector,

Use of adenovirus vector expressing the mouse full estrogen receptor alpha gene to infect mouse primary neurons

Estrogen plays important regulatory and protective roles in the central nervous system through estrogen receptor a mediation. Previous studies applied eukaryotic expression and lentiviral vectors carrying estrogen receptor a to cladfy the underlying mechanisms. In the present study, an adenovirus vector expressing the mouse full estrogen receptor a gene was constructed to identify biological characteristics of estrogen receptor a recombinant adenovirus infecting nerve cells. Primary cultured mouse nerve cells were first infected with estrogen receptor a recombinant adenovirus at various multiplicities of infection, followed by 100 multiplicity of infection. Results showed overexpression of estrogen receptor α mRNA and protein in the infected nerve cells. Estrogen receptor a recombinant adenovirus at 100 multiplicity of infection successfully infected neurons and upregulated estrogen receptor a mRNA and protein expression.

CXCL12 Retargeting of an Oncolytic Adenovirus Vector to the Chemokine CXCR4 and CXCR7 Receptors in Breast Cancer

Breast cancer is the most frequently diagnosed cancer in women under 60, and the second most diagnosed cancer in women over 60. While significant progress has been made in developing targeted therapies for breast cancer, advanced breast cancer continues to have high mortality, with poor 5-year survival rates. Thus, current therapies are insufficient in treating advanced stages of breast cancer;new treatments are sorely needed to address the complexity of advanced-stage breast cancer. Oncolytic virotherapy has been explored as a therapeutic approach capable of systemic administration, targeting cancer cells, and sparing normal tissue. In particular, oncolytic adenoviruses have been exploited as viral vectors due to their ease of manipulation, production, and demonstrated clinical safety profile. In this study, we engineered an oncolytic adenovirus to target the chemokine receptors CXCR4 and CXCR7. The overexpression of CXCR4 and CXCR7 is implicated in the initiation, survival, progress, and metastasis of breast cancer. Both receptors bind to the ligand, CXCL12 (SDF-1), which has been identified to play a crucial role in the metastasis of breast cancer cells. This study incorporated a T4 fibritin protein fused to CXCL12 into the tail domain of an adenovirus fiber to retarget the vector to the CXCR4 and CXCR7 chemokine receptors. We showed that the modified virus targets and infects CXCR4- and CXCR7-overexpressing breast cancer cells more efficiently than a wild-type control vector. In addition, the substitution of the wild-type fiber and knob with the modified chimeric fiber did not interfere with oncolytic capability. Overall, the results of this study demonstrate the feasibility of retargeting adenovirus vectors to chemokine receptor-positive tumors.

Down-regulation of Coxsakie and Adenovirus Receptor during Embryo Implantation

In this study,real-time PCR and immunohistochemistry were used to detect coxsakie and adenovirus receptor (CAR) expression.Both localization and quantity were evaluated in the uteri ob-tained at days post coitus (dpc) 2.5,4.5,6.5,8.5.Outcome of PCR was assessed by 2-ΔΔCt method.Im-age Pro-Plus 6.0 software was used for quantifying mean density of CAR expression in immunohisto-chemical sections.We found relatively weak CAR expression in the mouse uteri during implantation window.PCR and immunohistochemistry revealed highest CAR expression was detected on dpc 2.5 followed by down-regulation of CAR at dpc 4.5 and 6.5 (with significant difference).At dpc 8.5,CAR expression was increased slightly again.It is concluded that during implantation,the expression of CAR mRNA and protein is declined,resulting in the impairment of tight junction between cavity epithelium cells.After implantation window closure,CAR appears again to maintain epithelium stability.CAR might play an important role during embryo implantation procedure.

Adenovirus-mediated short hairpin RNA interference against p75 neurotrophin receptor in pheochromocytoma cells

Previous studies have confirmed that motor neuron apoptosis in the anterior horn of the lumbosacral spinal cord is positively correlated with p75 neurotrophin receptor （p75NTR） expression in rat models of cauda equina syndrome. This study used adenovirus to carry a short hairpin RNA （shRNA） for p75NTR gene silencing, to reduce p75NTR expression in the damaged phase and to decrease motor neuron apoptosis. Three p75 siRNA template oligonucleotide segments （shRNA） were designed, and cloned into the 1.0 CMV shuttle vector. HEK293 cells were cotransfected with shuttle vector （carrying shRNA） and an adenovirus vector framework expressing enhanced green fluorescent protein. Thus, this study successfully obtained adenovirus carrying p75shRNA. The obtained viruses were named Ad.shRNA1, Ad.shRNA2, and Ad.shRNA3. The recombinant adenoviruses were separately used to infect cultured pheochromocytoma cells （PC12）. Forty-eight hours later, p75NTR mRNA and total protein were analyzed from the PC12 cells. Compared with the negative controls, RNA interference rates were separately 98.49 ± 0.68%, 95.08 ± 1.79% and 96.60 ± 1.14% at the mRNA level, and 72.89 ± 2.17%, 58.83 ± 1.15% and 59.88 ± 0.44% at the protein level in the Ad.shRNA1, Ad.shRNA2, and Ad.shRNA3 groups, respectively. Thus, recombinant adenovirus shRNA-mediated gene silencing successfully suppressed p75NTR expression.

Inhibition of Metastatic Progression of SSTR2 Gene Transfection Mediated by Adenovirus in Human Pancreatic Carcinoma Cells

The inhibition of metastatic progression of Somatostatin receptor type 2 （SSTR2） gene transfection mediated by adenovirus in human pancreatic carcinoma cells and the mechanisms involved in this effect were studied. The full-length human SSTR2 cDNA was introduced into the pancreatic cancer cell line BXPC-3 by adenovirus-mediated transfection. Stable expression of mRNAs and protein of SSTR2 was detected by RT-PCR and Western-blot. The Matrigel-coated Transwell was used to detect the migratory and invasive ability of SSTR2-expressing cells, Adv-GFP control cells and mock control cells. Furthermore, the expression of matrix metalloproteinase-2 （MMP-2） and tissue inhibitor of metalloproteinase-2 （TIMP-2） was detected by RT-PCR in these cells. The stable expression of SSTR2 was detected in BXPC-3 transfected by Adv-GFP-SSTR2. A dramatic decrease of BXPC-3 expressing sst2 cells migrating through a Matrigel-coated filter was observed, as compared with Adv-GFP control and mock control cells （P〈0. 01）. Moreover, the expression of MMP-2 mRNA was significantly reduced in the SSTR2-expressing cells and converse- ly the expression of TIMP-2 mRNA was significantly increased in the SSTR2-expressing cells when compared with the Adv-GFP control and mock control （P〈0. 01）. The expression of reintroduced human SSTR2 gene in BXPC-3 cells by Adv-GFP-SSTR2 had the anti-migratory and anti-invasive effects, and the mechanisms involved in this effect may be due to the down-regulated expression of MMP-2 and up-regulated expression of TIMP-2.

重症腺病毒肺炎患儿血清IL-6R和MYD88水平与患儿预后的关系

目的分析重症腺病毒肺炎患儿血清白细胞介素-6受体(IL-6R)和髓样分化因子88(MYD88)水平与患儿预后的关系。方法将沧州市中心医院2020年6月至2022年6月收治的腺病毒肺炎患儿146例纳入研究,根据患儿病情严重程度分为非重症组(50例)和重症组(96例);根据重症腺病毒肺炎患儿出院时病情将患儿分为预后良好组(63例)及预后不良组(33例)。血清IL-6R和MYD88水平检测采用酶联免疫吸附法(ELISA)。分析重症腺病毒肺炎患儿血清IL-6R、MYD88水平及二者与序贯器官功能衰竭(SOFA)评分、Murray肺损伤评分、白细胞计数(WBC)、C反应蛋白(CRP)、血小板计数(PLT)的相关性。采用Logistic回归分析重症腺病毒肺炎患儿预后的影响因素。采用受试者工作特征(ROC)曲线分析血清IL-6R和MYD88水平对重症腺病毒肺炎患儿预后的预测价值。结果重症组血清IL-6R、MYD88水平及SOFA评分、Murray肺损伤评分均高于非重症组(P<0.05)。预后不良组WBC>10×10^(9)/L、CRP≥8 mg/L及PLT≥10×10^(9)/L患儿比例均高于预后良好组(P<0.05),IL-6R、MYD88水平及SOFA、Murray肺损伤评分均高于预后良好组(P<0.05)。血清IL-6R和MYD88呈正相关(P<0.05)。血清IL-6R、MYD88均与SOFA评分、Murray肺损伤评分、WBC、CRP以及PLT呈正相关(P<0.05)。经Logistic回归分析,IL-6R、MYD88升高是影响重症腺病毒肺炎患儿预后的危险因素(P<0.05)。经ROC曲线分析,血清IL-6R和MYD88联合预测重症腺病毒肺炎患儿预后的曲线下面积(AUC)优于各自单独预测(P<0.05)。结论重症腺病毒肺炎患儿血清IL-6R和MYD88水平显著升高,二者与患儿预后密切相关,二者联合较各自单独使用可更好地预测患儿预后。

石榴花水提物调节AHR/BNIP3改善糖尿病小鼠肝脏胰岛素信号

目的:探讨石榴花水提物(pomegranate flower water extract,PFW)对2型糖尿病小鼠肝脏胰岛素信号传导的影响及机制。方法:将C57BL/6J随机分为正常组、模型组、二甲双胍组(Met)、石榴花水提物低剂量组(PFWL)和石榴花水提物高剂量组(PFWH)。连续给药11周后,称小鼠体质量,检测空腹血糖(FBG)、胰岛素(INS)、甘油三酯(TG)和总胆固醇(TC)的含量,计算胰岛素抵抗指数(HOMA-IR);苏木素-伊红(HE)染色观察肝组织病理变化;Western blot法检测肝组织中胰岛素受体底物1(IRS1)、p-IRS1(Ser307)、蛋白激酶B(AKT)、p-AKT(Ser473)、糖原合成酶激酶-3β(Gsk3β)、p-Gsk3β(S9)、芳香烃受体(AhR)、磷脂酰乙醇胺N-甲基转移酶(PEMT)、Bcl-2/腺病毒E1B-19kDa相互作用蛋白3(BNIP3)蛋白表达。结果:与模型组比较,PFWH组FBG、INS、HOMA-IR、TG和TC含量极显著降低(P<0.01);PFWH组小鼠肝细胞内脂肪滴明显减少;PFWH组极显著升高肝脏中IRS1、p-AKT(Ser473)/AKT、p-Gsk3β(S9)/Gsk3β、BNIP3蛋白表达(P<0.01),极显著降低p-IRS1(Ser307)/IRS1、AHR、PEMT蛋白表达(P<0.01)。结论:PFW可能通过调节AHR/BNIP3抑制肝脏脂质沉积,改善p-IRS1(Ser307)/p-AKT(Ser473)/p-GSK3β(S9)胰岛素信号通路转导。

100例急性呼吸道感染儿童五项病毒联合检测结果分析

目的探讨100例急性呼吸道感染(ARI)儿童五项病毒联合检测结果。方法选取2022年1月至2022年12月我院收治的ARI患儿共计100例,所有患儿均接受肺炎支原体(MP)、肺炎衣原体(CP)、呼吸道合胞病毒(RSV)、腺病毒(ADV)、柯萨奇B组病毒(CVB)的免疫球蛋白M(IgM)抗体检测,分析五项病毒阳性检出率,并比较不同性别、不同年龄段、不同病情程度、不同发病季节患儿的阳性率。结果100例患儿中共38例患儿检出病毒抗体阳性(38.00%),其中单种病毒感染36例(36.00%),混合病毒感染2例(2.00%),所有感染类型中以MP感染居多(15.00%);不同性别患儿的五项病毒阳性率比较差异无统计学意义(P>0.05);1~6岁患儿与<1岁、>6岁患儿相比MP阳性率、总阳性率更高(P<0.05),<1岁患儿与>6岁患儿相比总阳性率更高(P<0.05),<1岁患儿与1~6岁、>6岁患儿相比RSV阳性率更高(P<0.05);重度患儿与轻度、中度患儿相比RSV阳性率、总阳性率更高(P<0.05);冬季发病患儿与春季发病患儿相比总阳性率更高(P<0.05)。结论MP是引发儿童ARI的主要病毒,五项病毒分布受到年龄、病情程度及发病季节影响,1~6岁患儿易受到MP感染,<1岁患儿易受到RSV感染,患儿病情加重多由RSV导致,且病毒性ARI在冬季高发。

血清TGR5 mRNA和BNIP3 mRNA在急性心肌梗死患者中的表达水平及临床意义

目的 探究血清G蛋白偶联胆汁酸受体5(TGR5)mRNA与Bcl-2/腺病毒QE1B-19kDa相互作用蛋白3(BNIP3)mRNA在急性心肌梗死(AMI)患者中的表达水平情况,以及二者对于术后心脏不良事件(MACE)发生的预测价值。方法 选取首都医科大学门头沟教学医院2018年1月至2020年1月收治的98例进行经皮冠状动脉介入术(PCI)的AMI患者[AMI患者包括急性非ST段抬高型心肌梗死(NSTEMI)46例与急性ST段抬高型心肌梗死(STEMI)52例]为研究组,另选取同期90例体检健康者作为对照组。采用实时荧光定量PCR(qRT-PCR)检测血清TGR5 mRNA和BNIP3 mRNA表达水平,根据PCI术后随访结果将研究组分为MACE组(46例)和非MACE组(52例)。收集两组患者临床资料,采用Pearson法分析术后MACE组血清TGR5 mRNA与BNIP3 mRNA表达水平的相关性,采用受试者工作特征(ROC)曲线分析血清TGR5 mRNA和BNIP3 mRNA对于术后AMI患者MACE发生的预测价值,采用Logistic回归分析AMI患者术后MACE发生的影响因素。结果 研究组血清TGR5 mRNA表达水平显著低于对照组,血清BNIP3 mRNA表达水平显著高于对照组,差异有统计学意义(P<0.05);术后MACE组左心室射血分数(LVEF)、TGR5 mRNA表达水平显著低于非MACE组,血清肌酐(SCr)、红细胞分布宽度(RDW)、Killip分级Ⅲ+Ⅳ级比例、BNIP3 mRNA表达水平显著高于非MACE组,差异有统计学意(P<0.05);经Pearson相关性分析,血清TGR5 mRNA与BNIP3 mRNA表达水平呈负相关(r=-0.543,P<0.05);ROC曲线分析显示,血清TGR5 mRNA预测AMI患者MACE发生的曲线下面积(AUC)为0.704(95%CI:0.601~0.808),血清BNIP3 mRNA预测AMI患者MACE发生的AUC为0.762(95%CI:0.696~0.883),血清TGR5 mRNA与BNIP3 mRNA联合预测AMI患者术后MACE发生的AUC为0.867(95%CI:0.783~0.932),优于二者单独预测(Z_(二者联合-TGR5)=2.346,Z二者联合-BNIP3=1.715,P=0.019、0.043);Logistic回归分析结果显示,TGR5 mRNA、BNIP3 mRNA、RDW是AMI患者术后MACE发生的影响因素(P<0.05)。结论 TGR5 mRNA在AMI患者血清中呈低表达水平,BNIP3 mRNA在AMI患者血清中呈高表达表达,二者对于预测术后MACE发生具有重要意义。

Overexpression of estrogen receptor beta alleviates the toxic effects of beta-amyloid protein on PC12 cells via non-hormonal ligands

After binding to the estrogen receptor, estrogen can alleviate the toxic effects of beta-amyloid protein, and thereby exert a therapeutic effect on Alzheimer＇s disease patients. Estrogen can increase the incidence of breast carcinoma and endometrial cancer in post-menopausal women, so it is not suitable for clinical treatment of Alzheimer＇s disease. There is recent evidence that the estrogen receptor can exert its neuroprotective effects without estrogen dependence. Real-time quantitative PCR and flow cytometry results showed that, compared with non-transfected PC12 cells, adenovirus-mediated estrogen receptor β gene-transfected PC12 cells exhibited lower expression of tumor necrosis factor a and interleukin 1β under stimulation with beta-amyloid protein and stronger protection from apoptosis. The Akt-specific inhibitor Abi-2 decreased the anti-inflammatory and anti-apoptotic effects of estrogen receptor β gene-transfection. These findings suggest that overexpression of estrogen receptor β can alleviate the toxic effect of beta-amyloid protein on PC12 cells, without estrogen dependence. The Akt pathway is one of the potential means for the anti-inflammatory and anti-apoptotic effects of the estrogen receptor.

sTREM1、ADA在腺病毒肺炎患儿血清中的表达及临床意义

目的探讨可溶性髓系细胞触发受体1(sTREM1)、腺苷酸脱氨酶(ADA)在腺病毒肺炎患儿血清中的表达及临床意义。方法选取沧州市中心医院2021年10月~2022年12月期间收治的97例腺病毒肺炎患儿作为腺病毒组,根据病情严重程度将腺病毒组患者分为轻型腺病毒肺炎组65例,重型腺病毒肺炎组32例,另取同期在沧州市中心医院体检,且一般资料与两组患儿相匹配的97例健康儿童作为对照组,Pearson法分析腺病毒肺炎患儿血清sTREM1和ADA表达水平的相关性;Logistic回归分析影响腺病毒肺炎严重程度的相关因素;受试者工作特征(ROC)曲线分析sTREM1和ADA对重型腺病毒肺炎的诊断价值。结果腺病毒组患儿血清sTREM1、ADA表达水平均显著高于对照组(P<0.05);腺病毒肺炎患儿血清sTREM1和ADA表达水平呈正相关(r=0.743,P<0.001);重型腺病毒肺炎患儿血清sTREM1和ADA表达水平均显著高于轻型腺病毒肺炎组(P<0.05);轻型和重型腺病毒肺炎组在既往有无肺部疾病史、PLT中比较,差异有统计学意义(P<0.05);既往有肺部疾病史、PLT≥300×10^(9)/L、sTREM1和ADA均为发生重型腺病毒肺炎的危险因素(P<0.05);血清sTREM1和ADA以及二者联合检测诊断重型腺病毒肺炎的AUC分别为0.599、0.790、0.930,二者联合检测优于血清sTREM1和ADA各自单独检测(Z二者联合-sTREM1=5.566、Z二者联合-ADA=4.860,P均<0.001)。结论血清sTREM1、ADA表达水平的升高与腺病毒肺炎的发生密切相关,二者联合检测对重型腺病毒肺炎具有较高的诊断价值。

Desmoglein 2(DSG2) Is A Receptor of Human Adenovirus Type 55 Causing Adult Severe Community-Acquired Pneumonia

Human adenovirus type 55(HAdV-B55) is a re-emergent acute respiratory disease pathogen that causes adult communityacquired pneumonia(CAP). Previous studies have shown that the receptor of HAdV-B14, which genome is highly similar with HAdV-B55, is human Desmoglein 2(DSG2). However, whether the receptor of HAdV-B55 is DSG2 is undetermined because there are three amino acid mutations in the fiber gene between HAdV-B14 and HAdV-B55. Here, firstly we found the 3T3 cells, a mouse embryo fibroblast rodent cell line which does not express human DSG2, were able to be infected by HAdV-B55 after transfected with pcDNA3.1-DSG2, while normal 3T3 cells were still unsusceptible to HAdV-B55 infection. Next, A549 cells with h DSG2 knock-down by siRNA were hard to be infected by HAdV-B3/-B14/-B55, while the control siRNA group was still able to be infected by all these types of HAdVs. Finally, immunofluorescence confocal microscopy indicated visually that Cy3-conjugated HAdV-B55 viruses entered A549 cells by binding to DSG2 protein.Therefore, DSG2 is a major receptor of HAdV-B55 causing adult CAP. Our finding is important for better understanding of interactions between adenoviruses and host cells and may shed light on the development of new drugs that can interfere with these processes as well as for the development of potent prophylactic vaccines.

Construction and identification of the recombinant adenovirus vector carrying a small interfering RNA targeting the peroxisome proliferator-activated receptor-γ

Background Steroid-induced osteonecrosis of the femoral head （ONFH） is a common clinical disease,with a high disability rate.At present,efficient prevention and treatment of steroid-induced ONFH is still lacking.The peroxisome proliferator-activated receptor-γ （PPARγ） is recognized as an important pathogenic gene for the development of steroid-induced ONFH.RNA interference （RNAi） is a tool for functional gene analysis,which has been successfully used to down-regulate the levels of specific target proteins.Therefore,down-regulation of PPARγ expression by RNAi may prevent the incidence of steroid-induced ONFH.Methods According to the principles of siRNA design,three duplex siRNA sequences （971-989,1253-1271 and 1367-1385） derived from the PPARy gene （NM_001082148） were synthesized.These duplexes were annealed,purified and ligated into 1.0-cytomegalovirus （CMV） shuttle vector.The shuttle vector was transfected into HEK293 cells.The HEK293 generated recombinant adenovirus vector carrying PPARγ siRNA sequences was purified and the titer of recombinant adenovirus was determined.Results After the annealing of single-strand DNA oligo encoding short hairpin RNA （shRNA） sequences,products were identified by gel electrophoresis.These products were ligated into the 1.0-CMV shuttle vector and the recombinant shuttle vectors 1.0-CMV-971,1.0-CMV-1253 and 1.0-CMV-1367 were constructed.These sequences of these recombinant vectors were confirmed.We then successfully constructed the recombinant adenovirus vector carrying siRNA targeting PPARγ.After purification,the virus titer was higher than 1010 plaque forming unit （PFU）/ml.Conclusion In this study,three recombinant adenovirus shuttle vectors carrying siRNA targeting PPARγ,including shuttle vectors 1.0-CMV-971,1.0-CMV-1253 and 1.0-CMV-1367,were successfully constructed and high titers of recombinant adenovirus were obtained.