1.Expression of tight junctions protein regulator PKC-θ in meningioma tissue
Guangchao ZHANG ; Xiaoqiu YANG ; Guofeng JIANG ; Yingwei LIU ; Chenshi QUAN
Journal of International Oncology 2010;37(3):228-229
Objective To study the expression and clinical meaning of tight junctions protein regulator PKC θ in human meningioma tissue.Method The expression of PKC θ in 30 non-invasion and 30 invasion meningioma tissue is tested by using immunohistochemistry S-P method,with the goal of analyzing the relationship between the expression of PKC θ and pathology character.Results The positive rate ofPKC θ in invasion meningioma tissue is obviously lower than in non-invasion meningioma tissue.The difference is significant(P<0.05)Conclusions The lower expression of PKC θ in invasion meningioma tissue pmbably affects occurrence and invasion of the meningioma by ehang4ng the structure and function of TJs.
2.TNFAIP8 gene silencing inhibits the migration of mouse RAW264. 7 macrophages
Fei YANG ; Suxia WU ; Shiming FENG ; Guangchao LIU ; Lihui CHAI
Chinese Journal of Microbiology and Immunology 2016;36(4):288-293
Objective To construct a shRNA lentiviral vector targeting the gene encoding tumor necrosis factor alpha-induced protein 8 (TNFAIP8) in RAW264. 7 cells, a mouse macrophage cell line, and to investigate the effects of TNFAIP8 gene silencing on the functions of mouse macrophages. Methods The shRNA sequence targeting TNFAIP8 gene was designed and DNA oligos containing small hairpin frame was synthesized. The double-stranded DNA was cloned into pLKO. 1-TRC vector after annealing. The recombi-nant vector was verified by using double enzyme digestion and gene sequencing. Lentiviruses were prepared by transfecting the constructed vector into 293T cells. Fluorescent quantitative RT-PCR and Western blot as-say were performed to detect the expression of TNFAIP8 at mRNA and protein levels after infecting the RAW264. 7 cells with lentiviruses. Flat dish adhesion experiment and wound-healing assay were used to evaluate the effects of TNFAIP8 gene silencing on the adhesion and migration of RAW264. 7 cells. Results The recombinant lentiviral vector was successfully constructed as indicated by double enzyme di-gestion and gene sequencing analysis. The expression of TNFAIP8 in RAW264. 7 cells at both mRNA and protein levels were significantly down-regulated after lentivirus infection (P<0. 05). Moreover, TNFAIP8 gene silencing significantly impaired the cell adhesion ability of RAW264. 7 cells after 15 min, 30 min or 2 hours of culture. Compared with the cells in control group, the RAW264. 7 cells harboring silenced TN-FAIP8 gene looked round with a smaller number of cellular extensions. The wound-healing assay showed that less TNFAIP8 gene-silenced RAW264. 7 cells migrated into the wounded area as compared with the cells in control group after 24 hours of culture (P<0. 05). The wound-healing rates of the experimental and control groups were 25% and 50%, respectively. Conclusion The recombinant lentiviral vector containing shRNA targeting the TNFAIP8 gene was successfully constructed. Transfecting the RAW264. 7 cells with the con-structed vector significantly silenced the expression of TNFAIP8 gene and inhibited the adhesion and migra-tion of these cells.
3.Synergistic effect of anti-human DR5 monoclonal antibody and adriamycin on SMMC-7721 cells
Jun ZHANG ; Yuanfang MA ; Guangchao LIU ; Shulian LI ; Feng LU
Chinese Journal of Immunology 1985;0(02):-
Objective:To determine the sensitivity of SMMC-7721 cells to mDRA-6,the synergistic damage effect of mDRA-6 and adriamycin on human hepatocellular carcimoma cell lines and its possible mechanism.Methods:SMMC-7721 cells were cultured with RPMI1640 medium in regular condition.The morphology was observed by microscope.Cytotoxicity was examined by MTT assay. Apoptosis were detected by flow cytometry.Results:(1)The apoptosis of SMMC-7721 cells could be induced by mDRA-6,1.89 ?g/ml of mDRA-6 cound kill 36% of the cells;(2)Concentration-dependent cytotoxicity of adriamycin was exhibited in SMMC-7721 cells;(3)The combination of mDRA-6 and adriamycin exhibited synergistic effect on SMMC-7721 cells.2 ?g/ml of mDRA6 and 40 ng/ml of adriamycin killed 60%SMMC-7721.Conclusion:MDRA-6 can induce SMMC-7721 cell apoptosis.The combination of mDRA-6 and adriamycin exhibit synergistic effect on SMMC-7721 cells.
4.Economic burden of patients with cervical cancer and precancerous lesions
Haitao LI ; Yan WANG ; Guqun SHEN ; Min YUAN ; Liping LIU ; Guangchao LIU ; Lin ZHU
Journal of Preventive Medicine 2022;34(4):350-356
Objective :
To investigate the economic burden of cervical cancer and precancerous lesions, so as to provide the evidence for improving the management of cervical cancer and formulating the policies for reducing the economic burden of cervical cancer and precancerous lesions.
Methods:
The hospitalized patients with cervical cancer and precancerous lesions were recruited from four hospitals in Xinjiang Uygur Autonomous Region from September 2020 to June 2021. The direct medical expenditures, direct non-medical expenditures, duration of absence from work in patients and their family members as carers were collected using a questionnaire designed by the Cancer Hospital of the Chinese Academy of Medical Sciences, and the economic burdens of cervical cancer and precancerous lesions were estimated. The factors affecting the economic burden of cervical cancer were identified using a multivariable linear regression model.
Results:
Totally 265 patients with cervical cancer and precancerous lesions were included, with an average age of ( 49.80±10.07 ) years. There were 170 patients with cervical cancer, including 64 cases with stage I, 79 cases with stage II, and 27 cases with stages III/Ⅳ, and 95 patients with precancerous lesions, including 33 cases with low-grade squamous intraepithelial lesion ( LSIL ) and 62 cases with high-grade squamous intraepithelial lesion ( HSIL ). The median economic burdens (interquartile range) were 11 481 ( 4 523 ), 17 850 ( 9 096 ), 112 883 ( 59 623 ), 150 875 ( 105 206 ) and 197 842 ( 61 844 ) Yuan per patient among cases with LSIL, HSIL, and stage I, II and III/Ⅳ cervical cancer, respectively, among which the direct medical expenditures accounted for 85.89% to 93.86%. The median economic burdens (interquartile range) were 708 ( 1 711 ), 11 678 (6 590), 2 557 ( 19 472 ), and 14 943 ( 27 773 ) Yuan per patient with precancerous lesions, and were 910 (1 530), 105 770 ( 91 019 ), 39 765 ( 30 490 ), and 146 445 ( 123 039 ) Yuan per patient with cervical cancer during the diagnostic phase, the clinical treatment phase, the follow-up phase, and in total, respectively. Multivariable linear regression analysis results showed that pathological stage ( β'=0.202, P=0.003 ) and duration of hospital stay ( β'=0.695, P<0.001 ) correlated with the economic burden among patients with cervical cancer.
Conclusion
There is a high economic burden among patients with cervical cancer and precancerous lesions. Advanced pathological stage and long duration of hospital stay may increase the economic burden among cervical cancer patients.
5.Perioperative complications of intertrochanteric fractures in patients aged 80 years and over
Guangchao ZHENG ; Haiping XU ; Huisong LIU ; Ping DONG ; Zhenxin LIU ; Xiaolei YE
Chinese Journal of Geriatrics 2016;35(5):530-532
Objective To investigate the prevention and management of perioperative complications of intertrochanteric fractures in elderly in-hospital patients(aged≥80 years).Methods Clinical data of 103 intertrochanteric fracture patients(31 male and 72 female)undergoing surgical treatment at our hospital from May 2010 to Nov.2015 were retrospectively analyzed.Their ages ranged from 80 to 99 years,with an average of 86.2 years.There were 3 Evan type Ⅰ cases,25 type Ⅱ cases,36 type Ⅲ cases,37 type Ⅳ cases and 2 type V cases.Of these,82 received epidural anesthesia and 21 had general anesthesia.Intertrochanteric fractures were treated with proximal femoral nail(PFN) internal fixation in 101 patients and dynamic hip screw(DHS)internal fixation in 2 patients.Results The average operation duration and blood loss were 30 min and 60 ml in the PFN internal fixation group and 60 min and 150 ml in the DHS internal fixation group,respectively.The average hospitalization time was 16.7 days.One patient (0.9%) died after operation,10 (9.7%) had preoperative complications of bed rest with 3 cases involving the central nervous system and 4 cases involving the respiratory system,and 38 cases (36.9%)had postoperative complications with 13 involving the central nervous system.The average time from admission to operation was 6 days,with 65 cases above the average and 38 cases below the average,and there was a statistically significant difference in the incidence of postoperative complications between the two subgroups(30/65 or 46.2 %for the former and 8/38 or 21.1% for the latter,x2=6.49,P<0.05).Conclusions Elderly patients with intertrochanteric fractures should undergo surgery as soon as possible,preferably with epidural anesthesia.Proficiency in surgical kills,short operation time,close monitoring of preoperative and postoperative conditions,and proper and timely treatment are the key to ensuring surgical success.
6.Reduced apoptosis induced by endotoxin in mice blocking TRAIL with soluble death receptor 5
Huiling BAI ; Xueyin WANG ; Shulian LI ; Hongying HUANG ; Yaowu DU ; Guangchao LIU ; Yuanfang MA
Chinese Journal of Microbiology and Immunology 2009;29(2):151-155
Objective To explore the effect of TNF related apoptosis inducing ligand (TRAIL) in apoptosis induced by LPS. Methods After LPS injected mice blocking TRAIL with soluble death receptor 5 (sDRS), detecting ALT, AST and LDH of mice serum at different times, apoptotic effects of LPS to mice hepatocyte were detected by HE and flow eytometry (FCM) with Annexin V-FITC/PI staining. The expres-sion of DR5 in mice hepatocyte was assayed with immunohistochemistry and FCM. Results Apoptotic effect was promoted by up-regulated DR5 expression on hepatocyte. Blocking TRAIL with sDR5 markedly amelio-rated the hepatocyte damage and reduced apoptosis. Conclusion These results establish a critical role for TRAIL in apoptosis during disease process of LPS.
7.Lung inflation with carbon monoxide during the cold ischemia phase ameliorates lung ischemia-reperfusion injury
Jiyu KANG ; Chao MENG ; Yang LIU ; Guangchao ZHANG ; Xue YANG ; Huacheng ZHOU
Chinese Journal of Organ Transplantation 2016;37(3):170-175
Objective To observe the effects and mechanism of lung inflation with carbon monoxide (CO) during the cold ischemia phase on lung ischemia-reperfusion injury (IRI) after rat lung transplantation.Method Twenty-four pairs of SD rats were selected to establish the model of lung transplantation,and random number method was used to divide 24 donors into 3 groups with 8 rats in each group.(1) CO inflation group (CO group):During the cold ischemia phase,500 ppm CO +volume fraction 40% O2 + N2 was used for lung inflation,and the volume was 5 mL/kg;(2) O2 inflation group (O2 group):During the cold ischemia phase,volume fraction 40% O2 + volume fraction 60% N2 was used for lung inflation;(3) Control group:The lung was deflated during the cold ischemia phase.The gas was replaced every 30 min in the CO and O2 groups,and the lung transplantations were performed after 180 min of cold ischemia.The arterial blood gas analysis was performed at baseline,3 min after reperfusion,and 60,120,and 180 min after reperfusion.The recipient serum levels of relative inflammatory factors,lung tissue cell apoptosis and nuclear factor kappa B (NF-κB) protein expression were detected after 180 min of reperfusion.Result As compared with the control group (238 ± 61 mm Hg),the oxygenation index in the O2 group (293 ± 78 mm Hg) and CO group (361 ± 48 mm Hg) was increased (P<0.05),and as compared with the O2 group,that in the CO group was increased (P<0.05).Furthermore,as compared with the control group,the interleukin (IL)-8,tumor necrosis factor (TNF)-α,and cell apoptosis in the O2 group and CO group were decreased significantly,and as compared with the O2 group,those in the CO group and NF-κB protein expression were significantly decreased (P<0.05).Conclusion Lung inflation with CO during the cold ischemia phase ameliorated the rat lung IRI via reducing the inflammatory response and cell apoptosis mediated by the NF-κB pathway.
8.NNAMB,a novel homospermidine conjugate,induces apoptosis and differentiation in B16 Melanoma cells
Songqiang XIE ; Yingliang WU ; Pengfei CHENG ; Minwei WANG ; Guangchao LIU ; Jin ZHAO ; Chaojie WANG
Chinese Pharmacological Bulletin 2003;0(10):-
0.1 ?mol?L-1), inducing differentiation through enhancement of melanogenesis and increase of the activity of tyrosinase at lower doses(
9.Eukaryotic expression and preparation of polyclonal antibody for osteoclast inhibitory lectin related protein
Suxia WU ; Lihui CHAI ; Zhanzheng WANG ; Guangchao LIU ; Wenzhi TIAN ; Yuanfang MA
Chinese Journal of Immunology 2014;(10):1374-1377,1392
Objective:In order to get recombinant protein OCILRP 2-Fc and anti-OCILRP2 antibody for further study of OCILRP2.Methods:Eukaryotic expression vector pIg-CD5-OCILRP2 which fused with extracellular domain of OCILRP 2 and human IgG1 Fc fragment was constructed.G418 was used for stable expression cell strain after pIg-CD5-OCILRP2 transfected into CHO cells.Recombinant protein OCILRP 2-Fc purified from CHO cell supernatant was used to immunize rabbit and anti-OCILRP2 polyclonal antibody was purified from rabbit serum by using protein G column.Results: ELISA data showed that we got a high-titer anti-serum and anti-OCILRP2 antibody purified from the rabbit serum.Western blot indicated this antibody could specifically bind to OCILRP 2-Fc and OCILRP2 in NIH/3T3 lysate.OCILRP2 expression in murine bone marrow derived dendritic cells ( DC) was detected by this polyclonal antibody ,too.Compared to immature DC ,OCILRP2 expression was elevated in LPS induced-mature DC.Conclusion: This study has offered an available tool and provided a clue for further study of the roles of OCILRP 2 in immune response.
10.Identification of biomarkers for esophageal squamous cell carcinoma based on bioinformatics
Yujun ZHANG ; Yan WANG ; Wusimanjiang Diliyaer ; Guangchao LIU ; Yanwu NIE ; Lin ZHU
Journal of Preventive Medicine 2022;34(9):906-913
Objective :
To identify biomarkers for esophageal squamous cell carcinoma (ESCC) using bioinformatics tools, so as to provide insights into diagnosis and targeted therapy of ESCC.
Methods:
The gene expression datasets GSE23400, GSE45670, GSE20347 and GSE17351 were screened and downloaded from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) of ESCC were screened using the online tool GEO2R, and the common DEGs among the four datasets were determined using Venn diagram. Gene Ontology (GO) annotations and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed using the DAVID database, and protein-protein interaction (PPI) analysis was performed using the STRING database. The key modules were identified using molecular complex detection (MCODE) plugin in the Cytoscape software, and hub genes with the highest connectivity degree were identified using the CytoHubba plugin, and the gene expression was validated on the UALCAN platform. Survival analysis of hub genes was performed using the Kaplan-Meier plotter database.
Results:
Totally 146 common DEGs were screened, including 102 up-regulated genes and 44 down-regulated genes. GO annotation analysis showed that the common DEGs were mainly enriched in biological processes of cell cycle, sister chromatid separation in the late mitotic phase and cell cycle regulation, enriched in cellular components of spindle and centrosome, and molecular functions of enzyme binding and ATP binding. KEGG pathway analysis showed that DEGs was significantly enriched in cell cycle, extracellular matrix (ECM)-receptor interactions and oocyte meiosis. A total of 10 hub genes were screened, and gene expression validation and survival analysis identified 7 genes associated with prognosis of ESCC, including CCNB1, CDK1, BUB1B, ZWINT, AURKA, MAD2L1 and MCM4, which were all highly expressed in ESCC specimens.
Conclusion
Seven hub genes of ESCC are identified based on bioinformatics, which may serve as biomarkers and therapeutic targets for ESCC.