Simultaneous unilateral central retinal vein occlusion and branch retinal artery occlusion after Coronavirus Disease 2019 (COVID-19) mRNA vaccine / Oclusão unilateral simultânea da veia retiniana central e artéria retiniana ramificada após vacina mRNA COVID-19

ABSTRACT A 51-year-old non-obese woman presented with a one-week history of progressive blurry vision within the inferior visual field of her left eye. Her only relevant past medical history was long-standing hypothyroidism and recent vaccination against Coronavirus Disease 2019 (COVID-19) with an mRNA vaccine 12 days before the onset of symptoms. At examination, the anterior segment was unremarkable, but the retinal fundus revealed a central retinal vein occlusion associated with a branch retinal artery occlusion of the superior temporal branch in her left eye. Ancillary tests to rule out thrombophilia, hyperviscosity, hypercoagulability, or inflammation were negative. Ultrasound tests were also negative for a cardiac or carotid origin of the branch retinal artery occlusion. At two-month follow-up, no new retinal vascular occlusive events were observed. Although the best-corrected visual acuity at presentation was 8/10 in the left eye, the final best-corrected visual acuity remained 3/10.

RESUMO Uma mulher de 51 anos, não obesa, apresentou história de uma semana de visão embaçada progressiva no campo visual inferior do olho esquerdo. Seu único histórico médico anterior relevante era hipotireoidismo de longa data e uma recente vacinação contra a Doença de Coronavírus 2019 (COVID-19), com vacina de mRNA, 12 dias antes do início dos sintomas. O exame mostrou segmento anterior normal, mas o fundo da retina revelou uma oclusão da veia central da retina associada a uma oclusão de ramo arterial da retina do ramo temporal superior no olho esquerdo. Testes auxiliares para descartar trombofilia, hiperviscosidade, hipercoagulabilidade ou inflamação apresentaram resultados negativos. Testes de ultrassom também foram negativos quanto a uma origem cardíaca ou da carótida da oclusão do ramo da artéria da retina. Após dois meses de acompanhamento, nenhum novo evento vascular oclusivo retiniano foi observado. Embora, a acuidade visual melhor corrigida na apresentação tenha sido de 8/10 no olho esquerdo, a acuidade visual final melhor corrigida permaneceu em 3/10.

Progress of expression and function of messenger RNA CCT6A in malignancies / 肿瘤研究与临床

Messenger RNA(mRNA) CCT6A can encode chaperone proteins and plays an important role in malignant tumors such as colorectal cancer, lung cancer and breast cancer. CCT6A is highly expressed in malignant tumors, which can be used as a biomarker to assess patients' prognosis, and promote malignant biological behaviors such as tumor proliferation and metastasis by regulating transforming growth factor β signals, cell cycles, and other pathways. CCT6A can also modulate immune cell infiltration in the tumor microenvironment and may be a potential target for tumor immunotherapy. The paper reviews the expression and function of CCT6A in malignancies.

Blood mRNA Expression Profiles of Autophagy, Apoptosis, and Hypoxia Markers on Blood Cardioplegia and Custodiol Cardioplegia Groups

Abstract Introduction: Blood cardioplegia (BC) and Custodiol cardioplegia (CC) have been used for a long time in open heart surgery and are highly effective solutions. The most controversial issue among these two is whether there is any difference between them regarding myocardial damage after ischemia surgery. In this study, autophagy, apoptosis, and hypoxia markers were investigated and that way we evaluated the differences between BC and CC patients. Methods: A total of 30 patients were included in this study, using two different cardioplegic solutions. Three different whole blood samples of the patients were taken from a central vein (preoperatively, immediately postoperatively, and one day after surgery). Total ribonucleic acid was extracted from these samples. Quantitative real-time polymerase chain reaction was performed, and changes in gene expression were determined by the 2-∆∆Ct method of relative quantification. Results: In the CC group, Beclin gene expression level was found to be higher and this difference was statistically significant (P=0.0024). Similarly, cysteine-aspartic acid protease (caspase) 9 and hypoxia-inducible factor 1α messenger ribonucleic acid (mRNA) gene expression level increased and were significantly different in the CC group. In the BC group, Beclin and microtubule-associated protein light chain 3 expressions were higher in the samples taken one day after surgery. Caspases 3 and 8 gene expressions were significantly different in the BC group. Conclusion: As a result of the analysis performed between the two cardioplegia groups, it has been shown that CC harms the myocardium more than BC at the level of mRNA expression of related markers.

Differentially expressed mRNA involved in the resistance of liver cancer to anlotinib / 临床肝胆病杂志

ObjectiveTo screen out the mRNAs involved in the resistance of hepatoma cells to anlotinib using ceRNA microarray. MethodsHigh-dose shock combined with low-dose induction was used to culture hepatoma cells resistant to anlotinib, and CCK8 assay was used to verify the difference in the proliferation of drug-resistant hepatoma cells treated by anlotinib. The ceRNA microarray was used to screen out the differentially expressed genes between drug-resistant hepatoma cells and normal hepatoma cells, and real-time PCR was used to verify the differentially expressed genes detected by some microarrays. the independent samples t-test was used for comparison of continuous data between two groups, and the Kaplan-Meier method was used to analyze the overall survival of hepatoma cells samples, and the log-rank test was used to compare survival rates. Fisher’s exact test was used for chip screening. ResultsThere was a significant difference in gene expression between drug-resistant hepatoma cells and normal hepatoma cells, and 10 genes with the greatest difference were screened out for analysis by reducing the range. There were 4 genes associated with drug resistance and tumor growth, i.e., BIRC2, BIRC7, ABCC2, and MAPK8. There were significant reductions in the expression levels of BIRC2, ABCC2, and MAPK8 (P=0001 4, 0001 2, and 0.011 8), and there was a significant increase in the expression of BIRC7 (P＜0.001). The results of real-time PCR were consistent with those of microarray (t=10.74,32.65,18.34, and 2.80; P=0.000 4, 0.000 1, 0.000 1, and 0.044 8). The high expression of BIRC7 and the low expression of MAPK8 were associated with the significant reduction in survival time (P=0.022 0 and 0.005 6). ConclusionBIRC2, BIRC7, ABCC2, and MAPK8 are differentially expressed between anlotinib-resistant hepatoma cells and normal hepatoma cells and may be involved in the resistance of hepatoma cells to anlotinib.

A bioinformatics analysis of the microRNA-mRNA differential expression network for alcoholic hepatitis / 临床肝胆病杂志

ObjectiveTo establish a microRNA-mRNA differential expression network for alcoholic hepatitis (AH), and to investigate new targets for the diagnosis and treatment of AH. MethodsDifferentially expressed microRNAs and mRNAs between AH patients and normal controls were screened out. Related software including TargetScan, DIANA, MIRDB, PICTAR, and miRWalk 2.0 was used to search for the target genes of differentially expressed microRNA, and a key microRNA-mRNA network was established using the differentially expressed mRNAs that changed in an opposite way to microRNA. The Database for Annotation, Visualization and Integrated Discovery was used for the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genome (KEGG) analyses of target genes. The GCBI online software (www.gcbi.com.cn) was used for enrichment analysis of target genes and core network establishment. The GeneMANIA database in Cytoscape software (genemania.org) was used to perform a protein-protein interaction analysis of key target genes. The above three methods were compared in terms of the search for key pathways involved in the development of AH. ResultsA key microRNA-mRNA network was established with 5 differentially expressed microRNAs including hsa-mir-21-5p, hsa-mir-148a-3p, and hsa-mir-30e-5p and 51 target genes including collagen type IV alpha 1 chain (COL4A1), thrombospondin-2 (THBS2), and integrin alpha 6 (IGTA6). A protein-protein interaction network of key target genes was established. The GO analysis and various pathway analyses showed that the PI3K-Akt pathway and local adhesion were closely associated with AH. ConclusionDuring the development of AH, there are complex interactions between the related proteins of key target genes. COL4A1 and THBS2 may promote the development of AH by activating ITGA6 to regulate the PI3K-Akt pathway and the process of local adhesion. The establishment of the microRNA-mRNA network reveals the key links in the development of AH and highlights the focus of research. The discovery of the genes associated with the PI3K-Akt pathway in AH is expected to provide new targets for the diagnosis and treatment of AH.

A bioinformatics analysis of the microRNA-mRNA differential expression network for alcoholic hepatitis / 临床肝胆病杂志

ObjectiveTo establish a microRNA-mRNA differential expression network for alcoholic hepatitis (AH), and to investigate new targets for the diagnosis and treatment of AH. MethodsDifferentially expressed microRNAs and mRNAs between AH patients and normal controls were screened out. Related software including TargetScan, DIANA, MIRDB, PICTAR, and miRWalk 2.0 was used to search for the target genes of differentially expressed microRNA, and a key microRNA-mRNA network was established using the differentially expressed mRNAs that changed in an opposite way to microRNA. The Database for Annotation, Visualization and Integrated Discovery was used for the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genome (KEGG) analyses of target genes. The GCBI online software (www.gcbi.com.cn) was used for enrichment analysis of target genes and core network establishment. The GeneMANIA database in Cytoscape software (genemania.org) was used to perform a protein-protein interaction analysis of key target genes. The above three methods were compared in terms of the search for key pathways involved in the development of AH. ResultsA key microRNA-mRNA network was established with 5 differentially expressed microRNAs including hsa-mir-21-5p, hsa-mir-148a-3p, and hsa-mir-30e-5p and 51 target genes including collagen type IV alpha 1 chain (COL4A1), thrombospondin-2 (THBS2), and integrin alpha 6 (IGTA6). A protein-protein interaction network of key target genes was established. The GO analysis and various pathway analyses showed that the PI3K-Akt pathway and local adhesion were closely associated with AH. ConclusionDuring the development of AH, there are complex interactions between the related proteins of key target genes. COL4A1 and THBS2 may promote the development of AH by activating ITGA6 to regulate the PI3K-Akt pathway and the process of local adhesion. The establishment of the microRNA-mRNA network reveals the key links in the development of AH and highlights the focus of research. The discovery of the genes associated with the PI3K-Akt pathway in AH is expected to provide new targets for the diagnosis and treatment of AH.

Research progress of lncRNA in the occurrence and metastasis of pancreatic cancer / 中华肝胆外科杂志

Pancreatic cancer is a malignant and gastrointestinal tumor.The five year survival for pancreatic cancer is very low.The pathogenic factors for pancreatic cancer are not yet clear.However,more and more studies have showed that lncRNAcontributed to pancreatic cancer proliferation and metastasis through genomic imprinting,chromatin modification,transcriptional activation and inhibition,nuclear transport,and protein function regulation.In recent years,with the continuous deepening of basic researches of pancreatic cancer,there were many long non-coding RNA deregulated in pancreatic cancer,which played an important role in tumororigenesis and metastasis.In this article,we reviewed the relevant researches on the mechanism of lncRNA in the occurrence and metastasis of pancreatic cancer.

Molecular Mechanism of Helicobacter pylori-associated Diseases Based on MiRNA-mRNA Regulatory Network / 胃肠病学

Background: Helicobacter pylori (Hp) infection is the leading cause of chronic gastritis, duodenal ulcer, and gastric cancer, and is associated with a variety of extragastrointestinal diseases. However, the molecular pathogenic mechanism of Hp and its interaction with the host are still unclear. Aims: To provide clues for understanding the interaction between Hp and host and the Hp-associated diseases through the establishment of a miRNA-mRNA regulatory network and bioinformatics analysis. Methods: Datasets from Gene Expression Omnibus (GEO) were used to determine the differentially expressed miRNAs and mRNAs in patients with Hp infection. Intersection of differentially expressed mRNAs and targeted mRNAs of the differentially expressed miRNAs was identified to form a miRNA-mRNA regulatory network. Gene ontology (GO)/KEGG enrichment analyses were performed, a protein-protein interaction network was constructed and the possible Hp-associated diseases were analyzed. Results: Gene set enrichment analysis revealed that the mostly enriched gene set was chemokine receptors bind chemokines. Using disease-related analysis of 28 core node genes in protein-protein interaction network, Hp infection was found to be associated with cancers and diseases of digestive, nervous, cardiovascular, respiratory and immune systems. Conclusions: Comprehensive analysis provides a method for predicting pathogen-related human diseases based on miRNA-mRNA regulatory network and points out that Hp infection might increase the risk of many extragastrointestinal diseases.

Screening and Regulatory Network Analysis of Core MiRNAs in Gastric Cancer Based on Bioinformatics / 胃肠病学

Background: MicroRNAs (miRNAs) are negative regulators of gene expression in various eukaryotes and play roles in RNA silencing and post-transcriptional regulation. Gene expression is affected by miRNAs dysregulation in almost all types of malignancies. Aims: To explore the core miRNAs regulatory network of gastric cancer, and to provide a theoretical basis for the analysis of molecular mechanism of miRNAs in the development of gastric cancer. Methods: MiRNAs and mRNA expression profile microarray were used to screen the differentially expressed miRNAs and mRNA. MiRWalk 2.0 was used to predict miRNAs-mRNA interactions, cross-matching with genes were selected by expression profile microarray to define the core differentially expressed miRNAs; and miRNAs-mRNA regulatory network was constructed. GO analysis and KEGG analysis were performed to analyze the targeted gene. Results: Twenty-one up-regulated miRNAs and 36 down-regulated miRNAs in gastric cancer were screened by expression profile microarray. After cross-matching, 1 042 low-expressed genes and 711 high-expressed genes were found, and 10 core miRNAs-mRNA regulatory networks were finally defined. The differentially expressed genes regulated by the core miRNAs were mainly involved in tumor-related pathways, and the high-expressed genes were mainly enriched in 9 signaling pathways such as ECM-receptor interaction, while the low-expressed genes were mainly enriched in 5 signaling pathways such as neuroactive ligand-receptor interaction. GO analysis showed that the up-regulated genes involved 315 functions and the down-regulated genes involved 88 functions, of which extracellular matrix organization was the most relevant. Conclusions: MiRNAs-mRNA regulatory network analysis based on bioinformatics provides a new perspective for gastric cancer research, which helps to systematically elucidate the molecular mechanism of miRNAs in the development of gastric cancer. It could provide a theoretical basis for the screening of biomarkers and the precise target selection for drug treatment of gastric cancer.

CK19 mRNA in peripheral blood combined with contrast-enhanced ultrasound: the predictive value of axillary lymph node involvement in breast cancer / 中华普通外科杂志

Objective To evaluate a model for axillary lymph node involvement combining CK19 mRNA with contrast enhanced ultrasound sonography (CEUS) score in operable breast cancer.Methods Operable breast cancer patients planned for sentinel lymph node (SLN) biopsy were enrolled.Preoperative CK19mRNA expressions in peripheral blood and CEUS score of axillary lymph nodes were tested before surgery.In the training set,postoperative sentinel lymph node (SLN) and non-sentinel lymph node (nSLN) pathological results were taken as the gold standard,effective modeling variables were screened,logistic regression was used to establish the prediction model.Parallel control studies were conducted between the validation set and the MSKCC model to evaluate the prediction accuracy and prediction efficiency.Results From Oct 2015 to Nov 2016,359 cases (training set) were enrolled and mathematical formulas for predicting SLN and nSLN were established,respectively.The sensitivity,specificity and AUC of predicting SLN involvement were 91.36％,94.92％ and 0.979 respectively.The sensitivity,specificity and AUC of predicting nSLN metastasis were 91.04％,90.53％ and 0.932 respectively.From Dec 2016 to Jul 2017,219 cases (verification set) were included.The sensitivity of SLN metastasis predicted by the model was 91.84％,the specificity was 96.69％,and the AUC was 0.979,significantly superior to the MSKCC model (0.739).The sensitivity,specificity and AUC of predicting nSLN metastasis were 95.35％,92.73％ and 0.945 respectively,significantly superior to the MSKCC model (0.873).Concolusions Combined with peripheral blood CK19 mRNA and CEUS score,the prediction model for axillary lymph node involvement for operable breast cancer,SLN/nSLN involvement probability can be calculated and qualitative judgment can be made.The overall accuracy and AUC of this model are better than the prediction model of MSKCC.

Expression of Toll-Like Receptor 4 and Matrix Metalloproteinase 8 in Gingival Crevicular Fluid in Patients with Periodontitis

Abstract Objective: To determine the expression of TLR4 and MMP8 in gingival crevicular fluid [GCF] in patients with periodontitis. Material and Methods: Clinical samples were collected from 23 gingival crevicular fluid of periodontal disease subjects (n = 14) and healthy periodontal subjects (n=9). Measurement of Clinical parameters of probing pocket depth (PPD), bleeding on probing (BOP), and clinical attachment loss (CAL) were included as diagnostic criteria. Pocket Depth (PD) and CAL were defined as present if the PPD was ≥ 4 mm and the CAL ≥ 1 mm. Expression of TLR4 and MMP8 in the gingival crevicular fluid of deep pockets (PD≥ 6mm), shallow pockets (PD 4-5 mm) and healthy periodontal sulcus (0-3 mm) were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Statistical analysis to compare the pocket was using Independent t-test and Mann-Whitney test. Correlation between mRNA expression and clinical parameters was analyzed using Spearman's correlation test Results: Expression of TLR4 was higher in shallow pockets compared to the control group, but the difference was not statistically significant (p>0.05). The expression of MMP8 was higher in shallow pockets compared to the control group, but the difference was not statistically significant (p>0.05) either. There is no significant correlation between TLR4 and MMP8 with clinical periodontal parameters Conclusion: TLR4 and MMP8 mRNA expression levels should not be used as a clinical biomarker in periodontitis diagnostic tools.

Expression analysis of long non-coding RNAs in a renal ischemia-reperfusion injury model

Abstract Purpose: To investigate the long non-coding RNAs (lncRNAs) profile on renal ischemia reperfusion in a mouse model. Methods: Microarray analysis was used to study the expression of misregulated lncRNA in a mouse model of renal ischemia reperfusion(I/R) with long ischemia time. Quantitative real-time PCR (qPCR) was used to verify the expression of selected lncRNAs and mRNAs.The potential functions of the lncRNA was analyzed by bioinformatics tools and databases. Results: Kidney function was impaired in I/R group compared to the normal group. Analysis showed that a total of 2267 lncRNAs and 2341 messenger RNAs (mRNAs) were significantly expressed in I/R group (≥2.0-fold, p < 0.05).The qPCR result showed that lncRNAs and mRNAs expression were consistent with the microarray analysis. The co-expression network profile analysis based on five validated lncRNAs and 203 interacted mRNAs showed it existed a total of 208 nodes and 333 connections. The GO and KEEG pathway analysis results showed that multiple lncRNAs are involved the mechanism of I/R. Conclusion: Multiple lncRNAs are involved in the mechanism of I/R.These analysis results will help us to further understand the mechanism of I/R and promote the new methods targeted at lncRNA to improve I/R injury.

Macrophage migration inhibitory factor as an incriminating agent in vitiligo

Abstract: Background: Vitiligo is an autoimmune skin disorder in which the loss of melanocytes is mainly attributed to defective autoimmune mechanisms and, lately, there has been more emphasis on autoinflammatory mediators. Among these is the macrophage migration inhibitory factor, which is involved in many autoimmune skin diseases. However, little is known about the contribution of this factor to vitiligo vulgaris. Objective: To determine the hypothesized role of migration inhibitory factor in vitiligo via estimation of serum migration inhibitory factor levels and migration inhibitory factor mRNA concentrations in patients with vitiligo compared with healthy controls. We also aimed to assess whether there is a relationship between the values of serum migration inhibitory factor and/or migration inhibitory factor mRNA with disease duration, clinical type and severity in vitiligo patients. Methods: Evaluation of migration inhibitory factor serum level and migration inhibitory factor mRNA expression by ELISA and real-time PCR, respectively, were performed for 50 patients with different degrees of vitiligo severity and compared to 15 age- and gender-matched healthy volunteers as controls. Results: There was a highly significant increase in serum migration inhibitory factor and migration inhibitory factor mRNA levels in vitiligo cases when compared to controls (p<0.001). There was a significant positive correlation between both serum migration inhibitory factor and migration inhibitory factor mRNA concentrations in vitiligo patients, and each of them with duration and severity of vitiligo. In addition, patients with generalized vitiligo have significantly elevated serum migration inhibitory factor and mRNA levels than control subjects. Study limitations: Small number of investigated subjects. Conclusions: Migration inhibitory factor may have an active role in the development of vitiligo, and it may also be a useful index of disease severity. Consequently, migration inhibitory factor may be a new treatment target for vitiligo patients.

Correlation between the expression of androgen receptor mRNA and the long-term tumor-specific and recurrence-free survival rate in patients with T1 bladder cancer / 国际外科学杂志

Objective To investigate the predictive value of androge receptors in stage pT1 non-muscle-invasive bladder cancer.Methods A total of 196 patients with stage pT1 non-muscle-invasive bladder cancer who underwent a transurethral resection of the bladder in Baoji People's Hospital from February 2003 to June 2012 were recruited to carry a retrospective analysis.The mRNA expression of the androge receptors transcript variants 1 (AR1) and 2 (AR2) was measured by quantitative real-time PCR.Spearman's correlation coefficient was used to analysis the correlation of androge receptors mRNA level to KRT5 and KRT20 mRNA.Results Kaplan-Meier analysis indicated that the recurrence-free survival,progression-free survival and cancer specific survival of patients with high AR1 mRNA expression (≥35.47) were significant better than patients with low AR1 mRNA expression (P ＜0.05).Multivariate COX regression analysis revealed that high AR1 mRNA expression was an independent prognostic marker for recurrence-free survival and cancer specific survival (P ＜ 0.05).Spearman rank correlation revealed a significant positive association between mRNA expression of AR1 and KRT5 (rs =0.3171,P ＜ 0.001) as well as a negative association with multifocal tumors (rs =0.1478,P ＜ 0.05).Conclusion Androge receptors mRNA expression can predict recurrence-free survival and cancer specific survival in patients with stage T1 nonmuscle-invasive bladder cancer.Further studies on the levels of androge receptors mRNA tend to be particularly important.

HER2 gene status and mRNA expression in immunohistochemistry 1+ breast cancer / 中华病理学杂志

Objective@#To investigate human epidermal growth factor 2 (HER2) gene status and in situ mRNA expression in breast cancers with immunohistochemistry(IHC) 1+ , and to reveal HER2 positive rate in these patients to provide reference data for obtaining precise HER2 results and modifying relevant clinical strategy to breast cancer.@*Methods@#Sixty-five IHC 1+ formalin-fixed and paraffin-embedded samples of invasive breast carcinoma of no special type (IBC-NST) were collected by surgical operation at Peking Union Medical College Hospital during 2011 to 2013. HER2 status and in situ mRNA expression were tested by fluorescence in situ hybridization (FISH) and RNAscope, respectively, by using tissue microarray. Metastatic lymph node was re-tested by FISH if HER2 status was equivocal or negative and with high expression of mRNA in the primary lesion.@*Results@#Four of 65 samples (6.2%) were FISH positive, which included 2 cases of HER2/CEP17>2 and average HER2 copy number>4 and 2 cases of HER2/CEP17<2 and average HER2 copy number>6. In the 4 samples of HER2 positive, 2 patients showed high in situ mRNA expression (3 scores by RNAscope), 2 patients showed moderate in situ mRNA expression (2 scores by RNAscope). In addition, 3 specimens with HER2/CEP17>2 and average HER2 copy number<4 were found in all patients, which included 2 cases of high in situ mRNA expression (3 and 4 scores by RNAscope) and 1 cases of moderate in situ mRNA expression (2 scores by RNAscope). There was no significant association between HER2 status or mRNA expression and clinicopathological characteristics, including tumor size, histopathological differentiation, lymph node metastasis and lymphovascular invasion (P>0.05).@*Conclusions@#A small number of HER2 IHC 1+ patients exist mRNA expression by using FISH method, which suggested that these patients might benefit from anti-HER2 therapy potentially. Since the importance for patients with breast cancers to develop diagnostic and therapeutic strategies from accurate molecular typing, further studies based on a larger cohort are needed to validate our findings.

Homogeneity of Different Functional mRNA Indicators for Wound Age Estimation / 法医学杂志

OBJECTIVES@#To explore the homogeneity level of four different functional mRNA （PUM2, TAB2, Cx45 and CHRNA1） expressions in rats with skeletal muscle contusion.@*METHODS@#The relative expressions of PUM2, TAB2, Cx45 and CHRNA1 mRNAs were detected by quantitative real-time reverse transcription-polymerase chain reaction （RT-qPCR）. The coefficient of variation （CV） of the relative expressions for different individuals in each injury group was calculated. The extreme value of CV, cumulative variability, and CVCV were compared.@*RESULTS@#A high CV of PUM2 and TAB2 mRNAs appeared on several different time points. However, the CV of Cx45 and CHRNA1 mRNAs was relatively low. The cumulative variability from high to low was PUM2, CHRNA1, TAB2 and Cx45 mRNAs. The relative expression of PUM2 mRNA was significantly higher than that of TAB2, Cx45 and CHRNA1 mRNAs （ P<0.05）. There was no statistical significance （P>0.05） in the CVCV of the relative expression of TAB2, CHRNA1 and Cx45 mRNAs.@*CONCLUSIONS@#As the mRNAs involving in biological process regulation, PUM2 and CHRNA1 mRNAs show a lowest individual homogeneity of the relative expression followed by TAB2 mRNA. As the mRNAs participating in the composition of cellular structure, Cx45 and CHRNA1 mRNAs show a high individual homogeneity of the relative expressions. The functional classification should be considered for the screening of the mRNA indicators used for wound age estimation.

DACT2 gene promoter area methylation status and mRNA expression in renal cell carcinoma / 重庆医学

Objective To explore the role of the DACT2 gene in the occurrence and development of renal cell carcinoma(RCC).Methods The samples of RCC tissues and corresponding tumor-adjacent tissues after radical operation and normal kidney tissues were collected.The methylation specific PCR (MSP) and real time fluorescence reverse transcriptase-PCR (RT-PCR) methods were adopted to detect the methylation status and mRNA expression of DACT2.The streptavidin-peroxidase (SP) method labeled by immunohistochemistry peroxidase was used to examine the expression of β-catenin protein.Then the relationship between DACT2 gene methylation status and mRNA expression with the clinicopathologic characteristics was analyzed.The relationship between DACT2 gene methylation with mRNA and β-catenin expression was analysed,as well.Results The DACT2 mRNA relative expression level in RCC tissues was 0.427±0.025,which was significantly lower than (0.801±0.047) in tumor-adjacent tissues and (0.872±0.022) in normal tissue,the positive rate of DACT2 gene methylation in RCC tissues was 45.76%,which was significantly higher than 6.78% in tumor-adjacent tissues and 5.08% in normal tissues,the difference was statistically significant (P0.05).The DACT2 gene mRNA expression level in RCC tissues and promoter area methylation occurrence rate had no obvious correlation with the clinical data such as patients age,gender,tumor size,clinical stage and Fuhrman grade (P>0.05).The DACT2 gene mRNA relative level in the methylation group was lower than that in the non-methylation group,the difference was statistically significant (P<0.05).The expression rate of β-catenin protein in cytoplasma in RCC tissues was higher than that in the tumor-adjacent tissues and normal tissues,the difference was statistically significant (P<0.05),moreover,DACT2 gen methylation had a positive correlation with β-catenin protein expression (r=0.324,P=0.012).Conclusion The decrease of DACT2 gene promoter area methylation and mRNA relative expression level may participate in the RCC occurrence,but has no relationship with RCC clinical progression.Methylation occurred in DACT2 gene promoter area may be one of reasons causing mRNA relative expression decrase.DACT2 gene methylation occurrence in RCC tissue might be related to the high expression of β-catenin.

Effect of dronedarone on HCN channel mRNA and protein expression in neonatal rat ventricular myocytes / 重庆医学

Objective To explore the effect of dronedaronel on hyperpolarization-activated cyclic-nucleotide-gated(HCN) channel expression by detecting the change of HCN channel mRNA and protein level before and after giving dronedarone in neonatal rat ventricular myocytes.Methods Neonatal rat ventricular myocytes were separated and digested by type Ⅱ collagenase,and then single ventricular myocytes were collected through differential sticking wall separation method.According to the concentrations (0.1,0.5,1.0,5.0,10.0,20.0 μmol/L of dronedaronel for treating myocytes for 48 h) and time(10 μmol/L of dronedaronel for treating myocytes for 1,6,12,24,48 h)the gradient grouping was conducted.The levels of HCN2 and HCN4 channel mRNA and protein level were determined by real-time PCR and Western blot.Results The HCN2 mRNA and HCN4 mRNA expression levels in concentration gradient group and time gradient group were lower than those in the control group(P＜0.05);compared with the control group,the protein level in the 10 umol/L dronedaronel treatment for 12 h group was significantly down-regulated(P＜ 0.01).Conclusion Dronedaronel could inhibit the expression of HCN2/HCN4 channel mRNA and protein,moreover its action shows the concentration dependency and reaches the maximum at 12 h after medication.

The effects of dynamic pressure on expression of Sox9 mRNA and protein in metaphyseal chondrocytes of rats / 天津医药

Objective To study the effect of dynamic stress stimulation on the expression of Sox9 mRNA and protein in metaphyseal chondrocytes in vitro, and to explore the specific mechanism of mechanical signal transduction. Methods The rat metaphyseal chondrocytes separated and cultured for the 3rd generation in vitro were randomly divided into four groups:control group (all interventions were not applied), simple dynamic pressure group (a dynamic pressure stimulus with a size of 90 mmHg and a frequency of 0.1 Hz was applied using an open pressure control culture system), simple calcium antagonist group (the concentration of 10μmol/L nifedipine was given) and dynamic pressure+calcium antagonist group (a dynamic pressure stimulus with a size of 90 mmHg, frequency of 0.1 Hz and concentration of 10 μmol/L nifedipine were given at the same time). The expression of Sox9 mRNA was detected after 24 h intervention by real-time quantitative polymerase chain reaction (RT-PCR) in four groups. The expression of Sox9 protein was detected by Western blot assay. The intracellular free Ca2+ in metaphyseal chondrocytes was labeled with Fluo-3/AM, and the average fluorescence intensity detected by laser scanning confocal scanning microscopy was compared between four groups. Results The expression of Sox9 mRNA was 3.81 times higher in dynamic stress group than that in the control group, and the protein expression level was 2.33 times higher than that of the control group (P<0.05). There were no significant differences in the expression of Sox9 mRNA and protein between the calcium antagonist group and the control group. The expressions of Sox9 mRNA and protein were lower in dynamic pressure+calcium antagonist group than those in the dynamic stress group, but which were higher than those of control group(P<0.05). The results of average fluorescence intensity showed that there was no significant difference in the intracellular free Ca2+ concentration between four groups (P > 0.05). Conclusion Dynamic stress stimulation can increase the expression of Sox9 mRNA and protein in rat metaphyseal chondrocytes. There is calcium channel involvement in the mechanical signal transduction.

The effects of dynamic pressure on expression of Sox9 mRNA and protein in metaphyseal chondrocytes of rats / 天津医药

Objective To study the effect of dynamic stress stimulation on the expression of Sox9 mRNA and protein in metaphyseal chondrocytes in vitro, and to explore the specific mechanism of mechanical signal transduction. Methods The rat metaphyseal chondrocytes separated and cultured for the 3rd generation in vitro were randomly divided into four groups:control group (all interventions were not applied), simple dynamic pressure group (a dynamic pressure stimulus with a size of 90 mmHg and a frequency of 0.1 Hz was applied using an open pressure control culture system), simple calcium antagonist group (the concentration of 10μmol/L nifedipine was given) and dynamic pressure+calcium antagonist group (a dynamic pressure stimulus with a size of 90 mmHg, frequency of 0.1 Hz and concentration of 10 μmol/L nifedipine were given at the same time). The expression of Sox9 mRNA was detected after 24 h intervention by real-time quantitative polymerase chain reaction (RT-PCR) in four groups. The expression of Sox9 protein was detected by Western blot assay. The intracellular free Ca2+ in metaphyseal chondrocytes was labeled with Fluo-3/AM, and the average fluorescence intensity detected by laser scanning confocal scanning microscopy was compared between four groups. Results The expression of Sox9 mRNA was 3.81 times higher in dynamic stress group than that in the control group, and the protein expression level was 2.33 times higher than that of the control group (P<0.05). There were no significant differences in the expression of Sox9 mRNA and protein between the calcium antagonist group and the control group. The expressions of Sox9 mRNA and protein were lower in dynamic pressure+calcium antagonist group than those in the dynamic stress group, but which were higher than those of control group(P<0.05). The results of average fluorescence intensity showed that there was no significant difference in the intracellular free Ca2+ concentration between four groups (P > 0.05). Conclusion Dynamic stress stimulation can increase the expression of Sox9 mRNA and protein in rat metaphyseal chondrocytes. There is calcium channel involvement in the mechanical signal transduction.