A cervical cancer tissue-derived decellularized extracellular matrix scaffold for cervical cancer tissue reconstruction in vitro / 南方医科大学学报

OBJECTIVE@#The prepare decellularized extracellular matrix (ECM) scaffold materials derived from human cervical carcinoma tissues for 3D culture of cervical carcinoma cells.@*METHODS@#Fresh human cervical carcinoma tissues were treated with sodium lauryl ether sulfate (SLES) solution to prepare decellularized ECM scaffolds. The scaffolds were examined for ECM microstructure and residual contents of key ECM components (collagen, glycosaminoglycan, and elastin) and genetic materials by pathological staining and biochemical content analysis. In vitro 3D culture models were established by injecting cultured cervical cancer cells into the prepared ECM scaffolds. The cells in the recellularized scaffolds were compared with those in a conventional 2D culture system for cell behaviors including migration, proliferation and epithelial-mesenchymal transition (EMT) wsing HE staining, immunohistochemical staining and molecular biological technology analysis. Resistance to 5-fluorouracil (5-Fu) of the cells in the two culture systems was tested by analyzing the cell apoptosis rates via flow cytometry.@*RESULTS@#SLES treatment effectively removed cells and genetic materials from human cervical carcinoma tissues but well preserved the microenvironment structure and biological activity of ECM. Compared with the 2D culture system, the 3D culture models significantly promoted proliferation, migration, EMT and 5-Fu resistance of human cervical cancer cells.@*CONCLUSION@#The decellularized ECM scaffolds prepared using human cervical carcinoma tissues provide the basis for construction of in vitro 3D culture models for human cervical cancer cells.

Mechanism of Gegen Qinliantang against Vulnerable Plaque of Atherosclerosis： Based on Macrophage Pyroptosis Mediated by NF-κB/NLRP3/Caspase-1 Pathway / 中国实验方剂学杂志

ObjectiveTo explore the effect of Gegen Qinliantang （GQL） on vulnerable plaque of atherosclerosis based on the macrophage pyroptosis mediated by nuclear factor （NF）-κB/NOD-like receptor protein 3 （NLRP3）/cysteine-aspartic acid protease （Caspase）-1 pathway. MethodA total of 12 normal C57BL/6CNC mice were used as the control group， and 60 ApoE-/- mice of the same line were randomized into 5 groups： model group， low-dose， medium-dose， and high-dose GQL groups （GQL-D， GQL-Z， GQL-G groups， respectively）， and western medicine group. The control group and model group were given （ig） equal volume sterile distilled， and GQL-D， GQL-Z， GQL-G and western medicine groups received （ig） corresponding concentration of drugs for 8 weeks. Aortic plaques were observed based on hematoxylin and eosin （HE） staining. Serum levels of interleukin （IL）-1β and IL-18 were detected by enzyme-linked immunosorbent assay （ELISA）， protein levels of macrophage mannose receptor （CD206）/apoptosis-associated speck-like protein containing a CARD （ASC） and CD206/NLRP3 by double-labeling immunofluorescence， and C-terminal gasdermin D （GSDMD）， N-terminal GSDMD， NLRP3， pro-cysteinyl aspartate specific proteinase 1 （pro-Caspase-1） and NF-κB p65 by Western blot. ResultCompared with the control group， model group demonstrated serious pathological changes， rise of the levels of serum IL-1β and IL-18 and tissue ASC， NLRP3， C-terminal GSDMD， N-terminal GSDMD， pro-Caspase-1， and NF-κB p65， and decrease of CD206 level （P<0.05）. As compared with model group， the administration groups showed alleviation of the lesions in aortic wall， decrease in levels of serum IL-1β and IL-18 and tissue ASC， NLRP3， C-terminal GSDMD， N-terminal GSDMD， pro-Caspase-1， and NF-κB p65， and rise of CD206 level， with significant difference between some groups （P<0.05）. ConclusionGegen Qinliantang alleviates vulnerable plaque of atherosclerosis by regulating NF-κB/NLRP3/Caspase-1 pathway and further relieving macrophage pyroptosis.

Gegen Qinliantang Regulates Polarization Tendency of Macrophages to Intervene in Vulnerable Plaque in AS of ApoE-/- Mice / 中国实验方剂学杂志

ObjectiveTo explore the mechanism underlying the intervention of Gegen Qinliantang （GQL） in vulnerable plaques in atherosclerosis （AS） of ApoE-/- mice by regulating the polarization of macrophages. MethodTwelve normal C57BL/6CNC mice were used as the control group， and 60 ApoE-/- mice of the same line were randomized into 5 groups： model group， low-dose， middle-dose， and high-dose GQL groups （GQL-D， GQL-Z， and GQL-G groups， respectively）， and atorvastatin group （western medicine group）. High-fat diet was used for modeling. The control group and the model group were given （ig） equal volume of sterile distilled water， and GQL-D， GQL-Z， GQL-G， and western medicine groups received （ig） corresponding concentration of drugs for 8 weeks. The levels of total cholesterol （TC）， triglyceride （TG）， high-density lipoprotein cholesterol （HDL-C）， and low-density lipoprotein cholesterol （LDL-C） were detected with biochemical methods. The distribution of plaques in the aortic region was observed based on oil red O staining and hematoxylin-eosin （HE） staining. Serum levels of M1 pro-inflammatory factors tumor necrosis factor （TNF）-α and interleukin （IL）-6 and M2 anti-inflammatory factors IL-13 and transforming growth factor （TGF）-β were detected by enzyme-linked immunosorbent assay （ELISA）. Protein expression of macrophage mannose receptor CD206/arginase-1 （Arg-1） and CD206/inducible nitric oxide synthase （iNOS） was determined by double-labeling immunofluorescence， and mRNA expression of aortic Arg-1 and iNOS by real-time polymerase chain reaction （PCR）. ResultLevels of TG， TC， and LDL-C were significantly lower and HDL-C level was significantly higher in the GQL-Z， GQL-G， and western medicine groups than in the model group. As the concentration of GQL rose， the area with plaques gradually shrunk and the color became lighter. The staining areas of the GQL-G group and the western medicine group were the most scattered. The administration groups showed significant increase in the protein levels of Arg-1 and CD206， significant decrease in the protein level of iNOS， significant rise of Arg-1 mRNA level， and significant drop of iNOS mRNA level （P<0.05）. ConclusionGQL intervenes in the vulnerable plaques in AS by improving lipid metabolism， inhibiting macrophage M1 polarization， promoting macrophage M2 polarization， and further improving the inflammatory microenvironment.

Explore Potential Molecular Mechanism of Gegen Qinliantang in Intervention of Atherosclerosis Based on Network Pharmacology and Molecular Docking / 中国实验方剂学杂志

ObjectiveThis study aims to explore the potential molecular mechanism of Gegen Qinliantang （GQL） in the intervention of atherosclerosis （AS） based on network pharmacology and molecular docking. MethodThe active components and targets of each medicinal in GQL were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， and AS-related genes from 7 databases. Thereby， the anti-AS targets of GQL were screened out. Cytoscape 3.8.0 was employed to construct the "component-target" network， and STRING the protein-protein interaction （PPI） network. Core targets were screened out with CytoNCA. R clusterProfiler was used for Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment of target genes， which were then visualized. Finally， molecular docking of the top ten active components with the core targets of AS was performed and the binding affinity was compared with that between atorvastatin and the core targets. ResultIn the end， 150 active components of GQL， 20 289 AS targets， and 213 common targets were retrieved， and 48 core common targets were screened out. They were mainly involved in the GO terms of nuclear receptor activity， ligand activation， and transcription factor activity and the pathways of fluid shear force and AS， advanced glycation end products-receptor for advanced glycation end products （AGE/RAGE）， interleukin-17 （IL-17）， tumor necrosis factor （TNF）， Toll-like receptor pathways and other signaling pathways closely related to AS. The molecular docking results showed that the effective components of GQL had high binding affinity to core targets of AS， and the binding affinity was even higher than that between the atorvastatin and core targets. The five groups with high binding affinity were puerarin-TNF， baicalein-inducible nitric oxide synthase 2 （NOS2）， puerarin-NOS2， and formononetin-NOS2， wogonin-NOS2. ConclusionThe above result provides new ideas for further exploration of this classical decoction.

Noninvasive indicators of indications for antiviral therapy in HBeAg-negative chronic HBV infection patients with alanine aminotransferase ≤40 U/L / 临床肝胆病杂志

Objective To investigate the noninvasive indicators of indications for antiviral therapy in HBeAg-negative chronic hepatitis B virus (HBV) infection patients with alanine aminotransferase (ALT) ≤40 U/L under the guidance of liver pathology. MethodsA retrospective analysis was performed for the clinical data of 377 HBeAg-negative chronic HBV infection patients with ALT ≤40 U/L who were hospitalized in Affiliated Hospital of Yan’an University, from October 2013 to August 2018 and underwent liver biopsy, among whom the patients with inflammatory activity ＜A2 and fibrosis stage ＜F2 were enrolled as non-antiviral therapy group(n=266), and the patients with inflammatory activity ≥A2 or fibrosis stage ≥F2 were enrolled as antiviral therapy group(n=111). The chi-square test was used for comparison of categorical data between two groups; the t-test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; univariate and multivariate binary logistic regression analyses were used to screen out the influencing factors for the initiation of antiviral therapy; the receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic efficiency of each indicator in determining the need for antiviral therapy in HBeAg-negative chronic HBV infection patients with ALT ≤40 U/L. ResultsOf all 377 patients, 266 (70.6%) did not need antiviral therapy for the time being, and 111 (29.4%) had marked liver damage and thus needed active antiviral therapy. The multivariate analysis showed that liver stiffness measurement (LSM) (odds ratio ［HR］=2.003, 95% confidence interval ［CI］: 1.647-2.437, P＜005), HBsAg (HR=1.563, 95% CI: 1.110-2.200, P＜0.05), HBV DNA (HR=1.519, 95% CI: 1173-1.966, P＜0.05), and albumin (HR=0.939, 95% CI: 0.884-0.998, P＜0.05) were independent influencing factors for the initiation of antiviral therapy. The ROC curve analysis showed that the area under the ROC curve (AUC) was 0.749 (95% CI: 0.699-0799) for LSM, 0642 (95% CI: 0.586-0.699) for HBV DNA, and 0.565 (95% CI: 0.507-0.623) for HBsAg, and the combination of LSM, HBV DNA, and HBsAg had a larger AUC of 0.779 (95% CI: 0.732-0.827). ConclusionThe levels of LSM, HBV DNA, and HBsAg have a reference value in determining the initiation of antiviral therapy in HBeAg-negative chronic HBV infection patients with ALT≤40 U/L.

Mechanism of Pinelliae Rhizoma-Citri Exocarpium Rubrum in Treatment of Metabolic Syndrome Based on Network Pharmacology / 中国实验方剂学杂志

Objective:By the method of network pharmacology， the mechanism of Pinelliae Rhizoma and Citri Exocarpium Rubrum in the treatment of metabolic syndrome was explored. Method:Effective components of Pinelliae Rhizoma and Citri Exocarpium Rubrum were retrieved by TCMSP database，and then selected by ADME parameters. TCMID，BATMAN-TCM，SymMap，TCM-MESH database were used to supplement effective components of TCMSP. TCMSP target prediction model was used to predict potential targets of drugs. DRUGBANK，DisGeNET，CTD，GeneCards，OMIM，PharmGkb，KEGG，DiGSeE databases were retrieved to obtain the targets of metabolic syndrome，and the chips were downloaded and analyzed through GEO database No.GSE98895 to screen out the differential genes of normal people and patients with metabolic syndrome，and supplement the target databases of metabolic syndrome. The intersections of Pinelliae Rhizoma-Citri Exocarpium Rubrum and metabolic syndrome disease targets were obtained by Rstudio 3.6.2. The above intersection targets were imported into the Metascape database for module analysis and overall GO（Biological Process），KEGG and Reactome pathway analysis. The core targets were selected from the intersection targets by using the cytohubba plug-in，the core genes were input into the database of BioGPS，Genecards to analyze the tissue distribution and subcellular distribution，and the core targets were assigned by using the database of DisGeNET. Result:A total of 34 active components and 120 targets of Pinelliae Rhizoma and Citri Exocarpium Rubrum were screened out，and 115 targets were obtained after the intersection of Rstudio 3.6.2. The results of Metascape module analysis and whole analysis were mostly the same，mainly involving the biological processes， such as ligand receptor interaction，calcium ion，cGMP-PKG，cholinergic synapse，thyroid hormone，insulin. The cytohubba plug-in was used to screen out 17 targets，involving 17 key genes， such as VEGFA and NOS3. The tissue and subcellular distribution of the core targets mainly included lymphoblasts，CD33+_myeloid cells，amygdala，pineal and cytoplasmic matrix，mitochondria. The main proteins were signal molecules，kinases and nucleic acids. Conclusion:Pinelliae Rhizoma and Citri Exocarpium Rubrum could treat metabolic syndrome through complex biological processes and pathways，such as blood circulation，ligand receptor interaction of nerve activity，cGMP-PKG，interleukin-related action，calcium ion. This indicates that traditional Chinese medicine（TCM） treated diseases through multi-component，multi-target，multi biological processes，multi-channel and other ways（which is also proved by the distribution of core genes in the tissue，subcellular and protein ascription information），indicating the superiority of the holism concept of TCM. Erchetang and its similar prescriptions are suitable for treating metabolic syndrome，which also indicates that the principle of "treating different diseases with the same therapy" of TCM is not only reflected at the theoretical level； and network pharmacology needs to be further proved in the combination with experimental verification.

Impact of smoking on young acute coronary syndrome patients evaluated by optical coherence tomography / 中国介入心脏病学杂志

Objective To explore the impact of smoking on coronary plaque characteristics on optical coherence tomography(OCT) in young patients with acute coronary disease(ACS).Methods We assessed the atherosclerotic plaque characteristics and vulnerability by OCT and coronary angiography in 60 ACS patients aged 45 years or younger in Beijing Anzhen Hospital, from June 2014 to June 2017. The patients were divided into the smoking group(n=33) and the non-smoking group(n=27) to compare the plaque characteristics and vulnerability.Results Smoking patients showed a less extent of fibrosis(48.55％vs. 77.8％,P=0.032)and microchannels(18.2％vs. 44.4％,P=0.033), and a greater extent of plaque rupture (24.2％vs. 3.7％,P=0.033) compared with non-smoking patients. In multivariate analysis, smoking was the only independent predictors of plaque rupture(OR 8.320, 95％CI 0.969-71.435,P=0.027) and less fibrosis (OR 0.269, 95％CI 0.086-0.837,P=0.020). Conclusions Patients who are smokers have less extensive fibrosis and a greater extent of plaque rupture, showing more extensive vulnerable plaque phenotype. Therefore, smoking is one of the major risk factors of advanced cardiovascular events in young patients.

The effects of group reminiscence therapy on depressive symptoms in institutionalized elderly: a Meta-analysis / 中国实用护理杂志

Objective To evaluate the effect of group reminiscence therapy on improving the depressive symptoms of the institutionalized elderly. Methods Collecting clinical randomized controlled trials and quasi-randomized controlled trials of group reminiscence therapy for improving depressive symptoms in institutionalized elderly which published from 2000 to November 2016, by searching the Cochrane Library, PubMed, CINAHL, PsycINFO, EMbase, China Biology Medicine, China National Knowledge Infrastructure, VIP, Wanfang Database, Taiwan Academic Online Database and Airiti Library. And reviewing the quality of the studies by using the tools for assessing risk of bias. RevMan 5.3 software was used for data analysis. Results Ten studies(2 randomized controlled trials and 8 quasi-experiments), including a total of 511 cases, were included. The results showed that group reminiscence therapy significantly improved the depression, life satisfaction, and self-esteem of institutionalized elderly (95%CI-1.11--0.56, 0.41-2.05, 1.99-3.82, all P<0.01). There was no statistically significant difference between group reminiscence therapy and usual nursing in relieving loneliness (P = 0.21). Conclusions Group reminiscence therapy can improve the depression in institutionalized elderly, decrease the degree of depression, improve the level of life satisfaction and self-esteem. But in the ease of loneliness has not been confirmed. The randomized controlled trials included in this study were rare and the quality of the literature was low, and large-scale randomized controlled trials of higher quality are needed to confirm this.

Emergency clinical laboratory system for civilian and military uses in actual military operations / 医疗卫生装备

Objective To develop an emergency clinical laboratory system for civilian and military uses to enhance medical support of filed medical unit for mass casualties.Methods The missions of field medical unit were analyzed in the actual confrontation,and an emergency clinical laboratory system for civilian and military uses was built to fulfill clinical laboratory support.The factors were explored for clinical laboratory examination,and some measures were taken accordingly.Results The requirements of field medical unit were met by the developed system.Conclusion The system can be applied in largescale military exercises to enhance clinical laboratory support ability of field medical unit.

Progress and challenges in optical cochlear implant / 中华耳鼻咽喉头颈外科杂志

Optical cochlear implant has been occuring as a new cochlear implant which utilizes laser pulses to stimulate hearing. Compared to electronic cochlear implant, it has demonstrated higher spatial selectivity and less radiation scattering, which could lead to higher fidelity cochlear prostheses. At present, most investigations have focused on experiments in vivo. Although a lot of exciting results have been obtained, the mechanisms of laser stimulation is still open. In this paper, a brief review on the recent new findings of optical cochlear implant is given, and possible mechanisms are discussed. In the end, new experimental proposals are suggested which could help to explore the mechanisms of laser-cochlea stimulation.

Diagnostic value of serum anti-phospholipase A2 receptor antibodies (PLA2R) in idiopathic membranous nephropathy / 中华检验医学杂志

Objective To investigate the diagnostic value and monitoring application of anti-phospholipase A2 receptor antibodies ( PLA2R ) in patients with idiopathic membranous nephropathy ( IMN) .Methods A retrospective study was used .48 patients were diagnosed as idiopathic membranous nephropathy by puncturing kidney from January of 2013 to June of 2014 in the Hospital of Liaoning Traditional Chinese Medical University and Shengjing Hospital of China Medical University were included. 43 patients were diagnosed as non-idiopathic membranous nephropathy ( NIMN ).35 healthy volunteers were selected as control groups.Serum PLA2R were detected by indirect immunofluorescence.The intensity of antibodies fluorescence and the level of urine protein in 24 hours(24 h PRO) ,serum UREA ,serum CYSC, serum UA,GFR were compared in each groups.Measurement data was shown as mean ±standard deviation;Count data was shown as frequency or composition, t test andχ2 test were used as statistical method.Results (1) Among 48 cases with IMN,37 cases showed positive PLA2R (positive rate 77.08%).All of cases were negative in NIMN group.The sensitivity and the specificity of serum PLA2R were 77.08%and 100%.(2) Compared with control groups, the levels of UREA, CYSC and UA were found statistically significant differences in patients with IMN ( P 0.05 ) . ( 3 ) Positive correlation were found between 24 h PRO(r=0.877,P=0.00),serum UA (r=0.766,P=0.00 ) with the level of PLA2R antibodies fluorescence intensity.( 4 ) There were no correlation between serum PLA2R antibody with IMN pathology aging (r=0.087,0.194,0.182;P=0.598,0.399,0.667).PLA2R positive rate(Ⅰstage:37.50%,Ⅱstage:84.85%, Ⅲstage 85.71%) may increases with the increasing of illness severity.Conclusion Serum PLA2R antibody would be a new laboratory diagnosis standard in patients with IMN.(Chin J Lab Med, 2015, 38:595-599 ) Objective To investigate the diagnostic value and monitoring application of anti-phospholipase A2 receptor antibodies ( PLA2R ) in patients with idiopathic membranous nephropathy ( IMN) .Methods A retrospective study was used .48 patients were diagnosed as idiopathic membranous nephropathy by puncturing kidney from January of 2013 to June of 2014 in the Hospital of Liaoning Traditional Chinese Medical University and Shengjing Hospital of China Medical University were included. 43 patients were diagnosed as non-idiopathic membranous nephropathy ( NIMN ).35 healthy volunteers were selected as control groups.Serum PLA2R were detected by indirect immunofluorescence.The intensity of antibodies fluorescence and the level of urine protein in 24 hours(24 h PRO) ,serum UREA ,serum CYSC, serum UA,GFR were compared in each groups.Measurement data was shown as mean ±standard deviation;Count data was shown as frequency or composition, t test andχ2 test were used as statistical method.Results (1) Among 48 cases with IMN,37 cases showed positive PLA2R (positive rate 77.08%).All of cases were negative in NIMN group.The sensitivity and the specificity of serum PLA2R were 77.08%and 100%.(2) Compared with control groups, the levels of UREA, CYSC and UA were found statistically significant differences in patients with IMN ( P 0.05 ) . ( 3 ) Positive correlation were found between 24 h PRO(r=0.877,P=0.00),serum UA (r=0.766,P=0.00 ) with the level of PLA2R antibodies fluorescence intensity.( 4 ) There were no correlation between serum PLA2R antibody with IMN pathology aging (r=0.087,0.194,0.182;P=0.598,0.399,0.667).PLA2R positive rate(Ⅰstage:37.50%,Ⅱstage:84.85%, Ⅲstage 85.71%) may increases with the increasing of illness severity.Conclusion Serum PLA2R antibody would be a new laboratory diagnosis standard in patients with IMN.

Effect of minimally invasive percutaneous plates versus interlocking intramedullary nailing in tibial shaft treatment for fractures in adults: A meta-analysis

OBJECTIVE: The aim of this article was to determine the effects of minimally invasive percutaneous plates versus interlocking intramedullary nailing in the treatment of tibial shaft fractures in adults. METHOD: Literature searches of the Cochrane Library, PubMed, EMBASE, the Chinese Biomedical Literature database, the CNKI database, Wanfang Data, and the Weipu Journal database were performed up to August 2013. Only randomized and quasi-randomized controlled clinical trials comparing the use of percutaneous plates and interlocking intramedullary nails for tibial shaft fractures were included. Data collection and extraction, quality assessment, and data analyses were performed according to the Cochrane standards. RESULTS: Eleven trials were included. Compared with interlocking intramedullary nailing, minimally invasive percutaneous plates shortened fracture healing time and resulted in lower rates of postoperative delayed union and pain. There was no significant difference between the two methods with regard to the rates of excellent and good Johner-Wruh scoring, the rate of reoperation, and other complications. CONCLUSIONS: Overall, insufficient evidence exists regarding the effects of minimally invasive percutaneous plates versus interlocking intramedullary nailing in the treatment of tibial shaft fractures in adults. Low-quality evidence suggests that minimally invasive percutaneous plates could shorten fracture healing time, decrease the rate of postoperative delayed union, and decrease pain levels compared with interlocking intramedullary nailing. There is no significant difference between the two groups in terms of functional recovery scores, reoperation, and other complications. Further research that includes high-quality randomized controlled, multicenter trials is required to compare the effects of minimally invasive percutaneous plates versus interlocking intramedullary nailing in the treatment of tibial shaft fractures in adults. .

Expression of recombinant human ZP3 protein using the baculovirus expression system / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the methods and solve the technical bottlenecks in the preparation of recombinant human protein hZP3 using the baculovirus expression system and pave the technical ground for the production and application of recombinant hZP3.</p><p><b>METHODS</b>The recombinant vector pFASTBAC HTa-hZP3 was constructed and transferred to competent E. coli cells carrying bacmid to produce recombinant bacmid by homologous recombination. Sf9 cells were transfected with the recombinant bacmid to produce recombinant baculovirus. Full-length recombinant hZP3 (amino acids 1-424) and truncated recombinant hZP3 (amino acids 23-348) were expressed in the sf9 cells by infection with the recombinant baculovirus. The expression time of hZP3 was determined by Western blot and its purification was explored.</p><p><b>RESULTS</b>The recombinant bacmid and baculovirus were successfully constructed for expressing both the full-length and truncated hZP3. The maximal expression of recombinant hZP3 in the sf9 cells was achieved at 72-96 hours after baculovirus infection. Some of the recombinant hZP3 with His-tag could bind affinity matrix and got purified but most of the solubilized hZP3 passed through and the reasons remained unknown. Purified recombinant hZP3 labeled with Dylight Dye488 was able to bind human sperm.</p><p><b>CONCLUSION</b>It is feasible to express recombinant hZP3 in insect cells using the baculovirus system though the yield of hZP3 needs to be optimized. The methods for efficient enrichment and purification of recombinant hZP3 require further exploration.</p>

Effect of silencing AEG-1 with small interfering RNA on the proliferation and cell cycle of gastric carcinoma SGC-7901 cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the effect of down-regulation of astrocyte elevated gene-1 (AEG-1) expression on cell proliferation and cell cycle of gastric carcinoma cells, and its possible molecular mechanism.</p><p><b>METHODS</b>Control siRNA and AEG-1 siRNA were transfected into gastric carcinoma SGC-7901 cells. 48 h after transfection, the cells were divided into 3 groups including untransfected, siRNA control and AEG-1 siRNA transfection groups. Expressions of AEG-1 mRNA and protein in the 3 group cells were detected by real-time quantitative PCR and Western blot. The changes of cell proliferation were examined using CCK-8 kit, and the cell cycle distribution was detected by flow cytometry. Finally, expressions of cell proliferation and cell cycle related proteins were detected by Western blot.</p><p><b>RESULTS</b>Real-time quantitative PCR and Western blot demonstrated that compared with the untransfected and siRNA control groups, expressions of AEG-1 mRNA and protein were significantly down-regulated in the AEG-1 siRNA transfection group (P < 0.05), but there was no significant difference between the untransfected and siRNA control groups (P > 0.05). Furthermore, in vivo experiment confirmed a significant down-regulation of AEG-1 protein in the AEG-1 siRNA transfection group (P < 0.05). In addition, AEG-1 siRNA obviously inhibited the proliferation of SGC-7901 cells at different time points after transfection with AEG-1 siRNA. The percentage of cells in G0/G1 phase in the AEG-1 siRNA transfection group [(61.26 ± 1.25)%] was significantly higher than those in the untransfected group [(46.17 ± 1.91)%] and siRNA control group [(46.46 ± 1.96)%], and there was a significant difference between them (all P < 0.001). Furthermore, the result of Western blotting revealed that down-regulation of AEG-1 expression evoked the down-regulation of cdk2 and cyclin D1 expressions and elevation of p21 expression in vitro and in vivo.</p><p><b>CONCLUSIONS</b>The inhibition of cell proliferation and cell cycle arrest mediated by down-regulation of AEG-1 expression may be closely associated with the changes of expression of cell cycle related proteins including cdk2, cyclin D1 and p21.</p>

Protective effect of cotransfection of A20 and HO-1 gene against the apoptosis induced by TNF-α in rat islets in vitro / 中华儿科杂志

<p><b>OBJECTIVE</b>To establish the method for cotransferring human A20 gene and human heme oxygenase-1 (HO-1) gene into the isolated rat islets using lentiviral transfection system, and to study the protective effect of A20 and HO-1 protein against the apoptosis induced by cycloheximide (CHX) and TNF-α, and finally to explore the underlying mechanism.</p><p><b>METHOD</b>The A20 gene and HO-1 gene were cloned and inserted into the lentiviral transfection system. The efficacy of gene transfer was measured by the intensity of the enhanced green fluorescent protein (EGFP) fluorescence-positive islets. Western blot was applied to verify the expression of the A20 and HO-1 genes. To induce apoptosis in vitro, the isolated islets were treated with CHX+TNF-α, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and the fluorescence-activated cell sorting (FACS) methods were used to evaluate the apoptosis of the islet cells and Western blot was used to detect caspase-3 activation.</p><p><b>RESULT</b>(1) A20 and HO-1 genes were introduced into the isolated islets by lentiviral transfection, both of the genes were highly expressed in the islets after 96 hours culture detected by Western blot method. (2) The insulin levels in the cell culture medium from A20 and/or HO-1 transgenic islets were significantly higher than that in non-transgenic controls (P < 0.01). (3)After CHX + TNF-alpha treatment, the cell culture medium insulin concentration in the A20 gene transfected group [(93.58 ± 4.12)µg/ml], HO-1 gene transfected group [(88.98 ± 4.77) µg/ml ] and A20/HO-1 co-transfected group [(103.33 ± 3.16) µg/ml] were significantly higher than that in the EGFP group [(9.03 ± 0.65) µg/ml ] and the control group [(8.86 ± 0.38) µg/ml] (P < 0.001). Minimum expression level of the activated caspase-3 was found in the A20/HO-1 co-transfected group.</p><p><b>CONCLUSION</b>The lentiviral gene transfer system was an efficient and stable gene transfer vector, the over-expressed A20 and HO-1 protein delivered via lentivirus could preserve rats' islets function and act against the apoptosis induced by CHX and TNF-α.</p>

Effects of KIAA0101 expression on proliferation and invasion of gastric carcinoma MKN-45 cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the expression of KIAA0101 protein in gastric carcinoma cells, and to explore the effects of its down-regulation on the cell proliferation, cell cycle and invasion.</p><p><b>METHODS</b>Western blot was used to detect KIAA0101 protein expression in three gastric carcinoma cell lines including MKN-28, SGC-7901 and MKN-45. KIAA0101 siRNA and control siRNA were utilized to transfect MKN-45 cells, respectively. CCK-8 was used to analyze the changes of cell proliferation, and flow cytometry to examine the changes of cell cycle distribution. Finally, Boyden chamber was used to detect the ability of cell invasion.</p><p><b>RESULTS</b>Relative level of KIAA0101 protein in MKN-45 cells was significantly higher than those in MKN-28 and SGC-7901 cells, and there was significant difference among the three cell lines (P < 0.05). The result of CCK-8 study demonstrated that, compared with untreated group and control siRNA group, the proliferation of MKN-45 cells in KIAA0101 siRNA group was significantly inhibited (P < 0.05). Additionally, the result of cell cycle analysis revealed that the percentage of cell number in G(0)/G(1) phase in KIAA0101 siRNA group [(61.47 ± 0.89)%] was significantly higher than those in untreated group [(47.43 ± 0.85)%] and control siRNA group [(48.43 ± 0.73)%; F = 271.653, P = 0.000]. Further, Boyden chamber assay showed that the cell numbers migrated to Matrigel in KIAA0101 siRNA group (61.51 ± 4.76) were significantly lower than those in untreated group (138.74 ± 10.16) and control siRNA group (132.93 ± 11.25; F = 65.949, P = 0.000).</p><p><b>CONCLUSIONS</b>Down-regulation of KIAA0101 expression leads to an inhibition of cell proliferation, cell cycle and cell invasion. It may provide a novel target for the treatment of patients with gastric carcinoma.</p>

Matrix metalloproteinase-2 and matrix metalloproteinase-9 mRNA expression in mice with collagen-induced arthritis following heterogenous umbilical cord blood stem cell transplantation / 中国组织工程研究

BACKGROUND: Matrix metalloproteinase (MMP) degrades extracellular matrix, which is a necessity of joint destruction in rheumatoid arthritis patients. MMP-2 and MMP-9 can evaluate rheumatoid arthritis and serve as an index to predict progressive destruction of the joint. OBJECTIVE: To observe heterogenous allogeneic umbilical cord blood stem cell (UBSC) transplantation on MMP-2 and MMP-9 expression in the spleen of mice with type Ⅱ collagen-induced arthritis. METHODS: Fetus cord blood was sterilely obtained and cord blood stem cells were separated. The C57BL/6(H-2b) mice were assigned to five groups (n=10). Except normal control group, models of collagen-induced arthritis were established using complete Freund's adjuvant + type Ⅱ collagen. Mice from the methopterin group were intragastrically administered methopterin suspension 0.017 5 g/kg, once every 5 days. Other groups used caudal vein injection. Mice from the model and normal control groups were injected with saline. Mice from the mono-UBSCs group and double-UBSCs group were injected with 2×106/kg UBSCs from one and two parents. At 42 days following injection, animals were sacrificed and the ankle joint was obtained for histopathological detection. MMP-2 and MMP-9 mRNA expression in the spleen was examined using reverse transcription-polymerase chain reaction. RESULTS AND CONCLUSION: Double-UBSC transplantation could significantly inhibit inflammatory cell infiltration in synovial tissue of mice with type Ⅱ collagen-induced arthritis, repaired impaired cartilage tissue. The repair effect was better than that in methopterin group and mono-UBSCs group. MMP-2 and MMP-9 mRNA expression in the spleen was significantly lower in the double-UBSCs group than the mono-UBSCs group (P < 0.01). These suggest that heterogenous allogeneic double-UBSCs transplantation participated in pathological changes in rheumatoid arthritis cartilage and in synthesis of cartilage extracellular matrix and effectively treated rheumatoid arthritis by regulating MMP-2 and MMP-9 mRNA expression.

Cochlear implantation and rehabilitation in prelingually deafened adolescents and young adults / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the outcomes of prelingually deafened adolescents and young adults receiving cochlear implantation (CI).</p><p><b>METHODS</b>Thirty-three patients with prelingual deafness aged 7-33 years at the time of CI were followed up for 4 years. The Speech Discrimination Score (SDS) was tested using disyllabic words and short sentences with close-set and open-set, respectively.</p><p><b>RESULTS</b>After appropriate mapping, sound field audiometry with warble tone showed that the warble tone threshold average (WTA) of the patients reached 25-41 dBHL, with a mean threshold of 28.62∓2.37 dBHL. At 1 year after the operation, the mean value of SDS was 70.03% in close-set word list with lip-reading and 61.18% without lip-reading, and was improved to 82.12% and 72.67% at 4 years, respectively. In open-set word list, the mean value of SDS was 77.94% with lip-reading and 67.85% without lip-reading. The safety and the benefits-cost evaluation of CI for prelingually deafened school age children and adolescents had been recognized by 90.9% of the families involved.</p><p><b>CONCLUSION</b>These patients can obtain appropriate speech discrimination scores with improved quality of life after CI.</p>

Intracerebroventricular transplantation of human amniotic epithelial cells ameliorates spatial memory deficit in the doubly transgenic mice coexpressing APPswe and PS1ΔE9-deleted genes / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Human amniotic epithelial cells (HAECs), which have characteristics of both embryonic and pluripotent stem cells, are therefore a candidate in cell therapy without creating legal or ethical problems. In the present study, we aimed to investigate the effects of intracerebroventricular transplantation of HAECs on doubly transgenic mice of Alzheimer's disease (AD) coexpressing presenilin-1 (PS1) and mutant Sweden amyloid precursor protein (APPswe) genes.</p><p><b>METHODS</b>The offspring mice genotypes were detected using PCR identification of APPswe and PS1 gene. The doubly transgenic (TG) mice (n = 20) and wild-type (WT) mice (n = 20) were randomly divided into two groups respectively: the transplantation group treated with HAECs and the control group with phosphate buffered saline. Six radial arm water maze test was used to assess the spatial memory in the TG and WT mice. Amyloid plaques and neurofibrillary tangles were analyzed using congo red and acid-silver methenamine staining respectively. Immunofluorescence cytochemistry was used to track the survival of HAECs. Immunohistochemistry was used to determine the expression of octamer-binding protein 4 (Oct-4) and Nanog in the HAECs. High performance liquid chromatography was used to measure acetylcholine in hippocampus. The density of cholinergic neurons in basal forebrain and nerve fibers in hippocampus was measured using acetylcholinesterase staining.</p><p><b>RESULTS</b>Amyloid deposition occurred in hippocampus and frontal cortex in the double TG mice aged 8 months, but not in WT mice. The results also showed that transplanted HAECs can survive for at least 8 weeks and migrate to the third ventricle without immune rejection. The graft HAECs can also express the specific marker Oct-4 and Nanog of stem cell. Compared with the control group, transplantation of HAECs can not only significantly improve the spatial memory of the TG mice, but also increase acetylcholine concentration and the number of hippocampal cholinergic neurites.</p><p><b>CONCLUSIONS</b>These results demonstrate that intracerebroventricular transplantation of HAECs can improve the spatial memory of the double TG mice. The higher content of acetylcholine in hippocampus released by more survived cholinergic neurites is one of the causes of this improvement.</p>

Clinical study of dynamic change of inflammatory factors in serum of acute paraquat rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the change of inflammatory factor in lung tissue of acute paraquat (PQ) poisoned rats.</p><p><b>METHODS</b>hundred SD rats were randomly divided into two groups: the normal control group (NC group, n = 10) and the PQ group (n = 80). The 1 ml saline was administered once in normal control group. The PQ group was administered with 25 mg/kg 1% PQ by intraperitoneal injection to establish the model of PQ induced renal injury. At six hours, at the first, the third and the seventh day the PQ group were sacrificed, while at the first day the normal control group was sacrificed. The level of normal tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), IL-6 in serum of rats were detected. Meanwhile, pathological changes of the renal were examined under optical microscope.</p><p><b>RESULTS</b>Histopathological findings of an earlier, a large number of patients edema clearly inflammatory cell infiltration. Compared with the control group, PQ exposure of serum TNF-α, IL-2, IL-6, the level at each time point were elevated. PQ treated group 6 h and 1, 3, 7 d when the IL-2 levels were (2.16 ± 0.65), (2.95 ± 1.02), (3.05 ± 1.12), (2.21 ± 0.62) µg/L, IL-6 were (62.5 ± 8.6), (85.6 ± 13.5), (90.3 ± 15.6), (65.3 ± 9.1) ng/ml, TNF-α were (1.95 ± 0.53), (2.86 ± 0.92), (3.15 ± 1.02), (2.06 ± 0.71) µg/L, compared with the control group, are significantly higher, the differences were statistically significant (P < 0.01).</p><p><b>CONCLUSION</b>acute PQ poisoning serum TNF-α, IL-2, IL-6 levels were significantly increased both early and late inflammatory factors involved in PQ poisoning the pathogenesis of renal injury.</p>