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Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the world, and the study on the regulatory mechanisms of the invasion and migration of HCC is of great significance to clinical diagnosis and treatment. Circular RNA (circRNA), as an important member of the non-coding RNA family, plays the role of microRNA (miRNA) sponge in hepatocytes due to its highly stable circular structure. It also plays an important role in HCC progression by regulating miRNA or promoting the expression of target genes through the competitive endogenous RNA mechanism. This article explores the mechanism of action of circRNA in the pathogenesis of HCC, so as to help with the screening for diagnostic markers of HCC and the development of effective therapeutic targets for HCC.
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Objective@#Apolipoprotein E (ApoE) is mainly synthesized in the liver. So far, it is unknown the relationship among APOE gene polymorphisms and WML, brain atrophy. Therefore, the aim of the study was to assess the associations of APOE gene polymorphisms in patients with WML and brain atrophy. @*Methods@#A total of 58 patients with WML, 128 patients with brain atrophy, 112 patients with co-occurrence of WML and brain atrophy and 95 healthy elderly volunteers were recruited from Renmin Hospital of WuHan University. @*Results@#Allele E3 was the most common allele. The alleles E2 had significantly higher levels of ApoB and lower age in WML group. The alleles E2 was associated with the lower level of ApoB, LDL-Ch, TCh, and sdLDL in co-occurrence group. The E3/E3 genotype has higher level of sdLDL, but lower age and female frequency in WML. The E3/E4 genotype had higher level of TG, but lower age in WML. Gender, Age, E2, Hyperhomocysteinemia and UA were also significantly associated with disease progression. @*Conclusion@#This study found that clinical data, lipids and metabolic complications were closely related to ApoE genotypes and alleles, and also disease progression and type.
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Objective:To investigate the positive rate for 2019-nCoV tests and co-infections in Wuhan district.Methods:A total of 8 274 cases in Wuhan were enrolled in this cross-sectional study during January 20 to February 9 in 2020, and were tested for 2019-nCoV using fluorescence quantitative PCR. Both respiratory tract samples (nasopharynx, oropharynx, sputum and alveolar lavage fluid) and non-respiratory tract samples (urine, feces, anal swabs, blood and conjunctival sac swabs) were collected. If both orf1ab and N genes are positive, they are classified as nucleic acid test positive group; if both orf1ab and N genes are negative, they are classified as negative group; if single gene target is positive, they are classified as suspicious group. Individuals were divided into male group and female group according to sex. At the same time, 316 patients were tested for 13 respiratory pathogens by multiplex PCR.Results:Among the 8 274 subjects, 2 745 (33.17%) were 2019-nCoV infected; 5 277 (63.77%) subjects showed negative results in the 2019-nCoV nucleic acid test; and 252 cases (3.05%) was not definitive (inconclusive result). The age of cases with COVID-19 patients and inconclusive cases was significantly higher than that of cases without 2019-nCoV infection (56>40, t=27.569, P<0.001; 52>40, t=6.774, P<0.001). The positive rate of 13 respiratory pathogens multiple tests was significantly lower in 104 subjects who were positive for 2019-nCoV compared with those in subjects who were negative for 2019-nCoV test (5.77% vs 18.39%, χ 2=24.105, P=0.003). Four types of respiratory tract samples and five types of non-respiratory tract sampleswere found to be positive for 2019-nCoV nucleic acid test. Conclusion:The 2019-nCoV nucleic acid positive rate inmale is higher than infemale. Co-infections should be pay close attention in COVID-19 patients. 2019-nCoV nucleic acid can be detected in non-respiratory tract samples.
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Objective@#To investigate the positive rate for 2019-nCoV tests and co-infections in Wuhan district.@*Methods@#A total of 8 274 cases in Wuhan were enrolled in this cross-sectional study during January 20 to February 9, 2020, and were tested for 2019-nCoV using fluorescence quantitative PCR. Both respiratory tract samples (nasopharynx, oropharynx, sputum and alveolar lavage fluid) and non-respiratory tract samples (urine, feces, anal swabs, blood and conjunctival sac swabs) were collected. If both orf1ab and N genes are positive, they are classified as nucleic acid test positive group; if both orf1ab and N genes are negative, they are classified as negative group; if single gene target is positive, they are classified as suspicious group. Individuals were divided into male group and female group according to sex. At the same time, 316 patients were tested for 13 respiratory pathogens by multiplex PCR.@*Results@#Among the 8 274 subjects, 2 745 (33.2%) were 2019-nCoV infected; 5 277 (63.8%) subjects showed negative results in the 2019-nCoV nucleic acid test; and 252 cases (3.05%) was not definitive (inconclusive result). The age of cases with COVID-19 patients and inconclusive cases was significantly higher than that of cases without 2019-nCoV infection (40 vs 56, t=27.569, P<0.001; 52 vs 56, t=6.774, P<0.001). The positive rate of 13 respiratory pathogens multiple tests was significantly lower in 104 subjects who were positive for 2019-nCoV compared with those in subjects who were negative for 2019-nCoV test (5.77% vs 18.39%, χ2=24.105, P=0.003). Four types of respiratory tract samples and five types of non-respiratory tract samples were found to be positive for 2019-nCoV nucleic acid test.@*Conclusion@#The 2019-nCoV nucleic acid positive rate in male is higher than in female. Co-infections should be pay close attention in COVID-19 patients. 2019-nCoV nucleic acid can be detected in non-respiratory tract samples.
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Objective@#To investigate the diagnostic value of immunoglobulin M (IgM) and immunoglobulin G(IgG) antibodies to 2019 Novel Coronavirus (2019-nCoV) in 2019-nCoV infection.@*Method@#This is a retrospective study. Serum samples were collected from 284 patients including outpatients and inpatients in the Renmin Hospital of Wuhan University from January 20, 2020 to February 17, 2020. Among them 205 cases were 2019-nCoV infected patients, including 186 cases confirmed with nucleic acid test and 19 cases diagnosed by clinical symptoms and CT characteristics according to "the New Coronavirus Pneumonia Control Protocol (5th edition)" . A total of 79 subjects with other diseases but negative to 2019-nCoV infection were recruited as control group. Serum IgM and IgG antibodies to 2019-nCoV were measured with fully automated immunoassay technology for all subjects. Statistical significance between 2019-nCoV antibodies test and 2019-nCoV nucleic acid test was determined using the χ2 tests.@*Result@#The sensitivity of serum IgM and IgG antibodies to 2019-nCoV were 70.24%(144/205) and 96.10%(197/205) respectively and the specificity were 96.20%(76/79) and 92.41%(73/79) respectively. The positive and negative predictive values of 2019-nCoV antibodies were 95.63%(197/206) and 91.03% (71/78) respectively, and the positive and negative predictive values of 2019-nCoV nucleic acid test were 100%(186/186) and 80.61%(79/98) respectively. The total coincidence rate of diagnosing 2019-nCoV infection between antibody tests and nucleic acid test for 2019-nCoV were 88.03%(250/284).@*Conclusion@#Joint detection of serum IgM and IgG antibodies to 2019-nCoV is an effective screening and diagnostic indicators for 2019-nCoV infection, and an effective complement to the false negative results to nucleic acid test.
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In December, the outbreak of a novel coronavirus (2019-nCoV) in Wuhan, China, has attracted extensive global attention. On January 20, 2020,the Chinese health authorities upgraded the coronavirus to a Class B infectious disease in the Law of the People's Republic of China on the Prevention and Treatment of Infectious Diseases, and considered it as Class A infectious diseases in disease control and prevention. On January 22, 2020, the 2019-nCoV nucleic acid detection test was listed as the diagnostic criteria in the "guidelines for diagnosis and treatment of pneumonia due to 2019-nCoV (Trial Version 2)" . Therefore, standardizing the operation process of the 2019-nCoV nucleic acid detection in clinical laboratories has become a top priority. It is of paramount importance to establish standard protocols for detection of the 2019-nCoV nucleic acids in clinical laboratories to improve the reliability of the results and ensure the biosafety of laboratory personnel.
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Hepatitis C virus (HCV) infection is one of the leading causes of liver cirrhosis, end-stage liver disease, and even liver cancer. Patients with HCV infection tend to have insulin resistance and dyslipidemia, which may lead to a series of metabolic syndromes and greatly threaten human health. At present, there are still no effective vaccines to prevent HCV infection, and therefore, the mechanism of HCV infection remains a hot topic in clinical research. Many studies have shown that there is a complex relationship between HCV infection and lipids, but the role of lipids in HCV infection remains unclear. This article reviews the current research status of the role of lipid metabolism in HCV infection and life cycle, in order to understand the mechanism of abnormal fat metabolism in the liver and the pathogenesis of fatty liver disease.
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ObjectiveTo investigate the influence of hepatitis C virus (HCV) on the expression of lipid metabolism indices in vitro and in vivo. MethodsA total of 114 samples of patients with HCV infection who were treated in Renmin Hospital of Wuhan University from September 2017 to September 2018 were collected as experimental group, and 96 samples of healthy individuals who underwent physical examination were collected as control group. An automatic biochemical analyzer was used to measure blood lipid parameters, including triglyceride (TG), total cholesterol (TCh), high-density lipoprotein (HDL), low-density lipoprotein (LDL), small and low-density lipoprotein (sdLDL), lipoprotein (a), apolipoprotein A1 (ApoA1), and apolipoprotein B (ApoB). RT-qPCR was used to measure the mRNA expression of fatty acid synthase (FASN), acetyl-CoA carboxylase (ACACA), hydroxymethylglutarate mono-acyl CoA reductase (HMGR), ApoA1, ApoB, and low-density lipoprotein receptor (LDLR) in Huh7.5.1 cells with or without HCV infection. 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. ResultsCompared with the control group, the experimental group had significantly lower serum levels of TCh (2.98±0.51 mmol/L vs 4.24±0.43 mmol/L, t=4.96, P＜0.05), HDL (0.87±0.16 mmol/L vs 1.24±0.21 mmol/L, t=5.65, P＜0.05), LDL (1.75±0.24 mmol/L vs 2.64±0.37 mmol/L, t=3.88, P＜0.05), ApoA1 (0.94±0.18 mmol/L vs 1.47±0.26 mmol/L, t=3.71, P＜0.05), and ApoB (0.67±0.31 mmol/L vs 0.98±0.14 mmol/L, t=4.41, P＜0.05). Huh7.5.1 cells with HCV infection had significantly lower mRNA expression of ApoA1 than those without HCV infection (t=-3.43, P＜0.05), as well as significantly higher mRNA expression of FASN, ACACA, HMGR, ApoB, and LDLR (t=5.40, 4.93, 3.34, 6.88, and 3.84, all P＜0.05). ConclusionHCV infection can upregulate the mRNA levels of enzymes involved in the synthesis of fatty acids and cholesterol and thus affect lipid metabolism in vivo and in vitro.
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Objective To explore serum liver and kidney function,blood lipid,complement indicators in patients with hepatitis B-associated nephropathy and the relationship between HBV infection and kidney function.Methods 141 cases with hepatitis B-associated nephropathy were randomly selected from January 2015 to July 2016 from Renmin Hospital of Wuhan University including 96 male,female 45 eases.65 cases with nephropathy included 31 male,female 34 eases.68 cases with healthy people included 36 male and female 32 cases.Liver and kidney function parameters were detected by the automatic biochemical analyzer Siemens ADVIA 2400.Complement C3 and C4 were detected by the automated immunoassay analyzer Beckman 1MMAGE 800.HBV DNA load was detected by fluorescence quantitative PCR instrument ROCHE LightCycler(R)480.Results ALT,Cr,UA,Urea,β2-M,LDL,TCh,TG,C3 and C4 were significant difference among hepatitis B complicated with nephropathy,nephropathy group and healthy groups (FALT =6.50,FCr=46.02,FUA =32.89,FUrea =37.60,Fβ2 M =44.98,FLDL=4.13,FTch =5.20,FTG =26.90,FC3 =14.54,Fc4 =11.01,all P＜0.05).ALT,AST,β2-M,C3 and C4 were significant difference among hepatitis B activity period complicated with nephropathy,hepatitis B non-activity period complicated with nephropathy and nephropathy group (FALT =5.96,FAST =7.45,Fβ2-M =18.70,FC3 =5.32,Fc4 =4.16;all P＜0.05).Conclusion Concentrations of liver and kidney function,blood lipid abnormalities and complement had a certain relationship with the hepatitis B complicated with nephropathy and HBV had a certain effect on serum concentrations of β2-M and complement.
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High mobility group box 1 protein,a kind of damage-associated molecular patterns,is a group of nonhistone DNA-binding protein in the nuclear.It not only involves in DNA replication,recombination,repair,regulation of gene transcription,but also induces activation of innate immunity and T cell generation and activates the adaptive immune response through Toll-like receptor.It is also more closely related to autoimmune diseases,inflammatory diseases,tumors and so on.Currently there are many researches on the role of TLR-mediated HMGB1 signaling pathway in diseases,which is involved in a variety of pathological processes and has a broad clinical outlook.
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Objective To detect mRNA expressions of STING and type Ⅰ interferons(IFN-α and IFN-β) in peripheral blood mononuclear ceils (PBMCs) of patients with chronic hepatitis C(CHC).Methods 113 patients with CHC and 94 healthy controls were collected from Renmin Hospital of Wuhan University during January 2015 and January 2016.Expressions of mRNA of STING,RIG-1,IFN-α and IFN-β were detected by real-time quantitative PCR,and their relative expression values were obtained by 2-△△Ct method and the results were statistically analyzed.Results The expression of mRNA of STING,RIG-1,IFN-α and IFN-β in CHC were 0.018,0.361,0.578 and 0.573 times than its in healthy controls respectively and the differences were statistically significant (t-8.28,4.26,2.18 and 2.07,all P ＜ 0.05).Pearson correlation analysis showed that mRNA expressions of STING in healthy controls were positively correlated with that of IFN-α mRNA and IFN-β mRNA (r=0.487 and 0.207,all P＜0.05),which had no correlation in CHC (r=0.174 and 0.091,all P＞0.05).The expressions of STING mRNA was positively correlated with that of RIG-1 mRNA in healthycontrols (r=0.222,P＜0.05),but there was no correlation of expression in CHC between STING mRNA and RIG-1 mRNA (r=-0.029,P＞0.05).Conclusion Compared with healthy controls,the expression of STING,RIG-1,IFN-α and IFN-β mRNA were all reduced.There was positive correlation between STING and RIG-1,IFN-α and IFN-β in healthy controls,but no correlation in CHC.
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Objective To detect the expression of retinoic acid-inducible gene Ⅰ ( RIG-Ⅰ) in peripheral blood mononuclear cells (PBMCs) of patients with chronic hepatitis C (CHC), and to explore its correlations with type Ⅰinterferon gene expression and serum level of hepatitis C virus .Methods A total of 101 na?ve patients with CHC were enrolled from Renmin Hospital of Wuhan University during July 2014 and July 2015, and 69 healthy subjects served as controls .The expressions of RIG-Ⅰ, IFNαand IFNβmRNA in PBMCs and serum level of HCV RNA were detected by real-time quantitative polymerase chain reaction.Pearson correlation analysis was performed to investigate the correlations of RIG-Ⅰ mRNA expression with IFNα/βmRNA expression and serum level of HCV RNA .Results The relative expressions of RIG-Ⅰ, IFNα, IFNβmRNA to healthy controls in CHC group were 0.082, 0.022 and 0.156, respectively (t=7.83, 3.65 and 2.13, all P0.05).Conclusion Expression of RIG-ⅠmRNA decreases in patients with CHC , and is positively correlated with that of IFNαand IFNβmRNA.
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Hepatitis C virus (HCV)was easily developed into hepatitis,cirrhosis and liver cancer after infecting human.Cur-rently,the traditional method of treatment of HCV was pegylated interferon and ribavirin program,which was ineffective and had poor side effects,while the new direct antiviral drugs were expensive.Therefore,looking for cheap and non-toxic side effects of treatment was the focus of current research.More and more evidence indicate that micro-Ribose Nucleic Acid (miRNAs)play an important role in the development of liver disease,regeneration and functional regulation.This review studies the relationship between miR-122 and Hepatitis C virus and hepatocellular carcinoma.
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Objective To investigate the association of rs10004195 single nucleotide polymorphisms in Toll like re-ceptor 10(TLR 10) gene with helicobacter pylori infection and associated diseases. Methods A total of 652 pa-tients who has been examined by gastroscopy were obtained, and then peripheral blood samples were collected from all the patients. Immune colloidal gold method was used to test the serological Hp antibody of all the patients. The TLR10 gene rs10004195 polymorphisms were examined by direct DNA sequencing of the PCR products. Results The frequencies of AA,TT and AT genotype on TLR10 rs10004195 were 30. 98%, 20. 71%, 48. 31%;there was significant difference between Hp antibody positive group and Hp antibody negative group in the TT frequencies of TLR10 rs10004195 ( P<0. 05 ) . No significant difference between controls and Hp associated diseases groups in Hp antibody positive group or in Hp antibody negative group were observed. Conclusion There was correlation be-tween the TLR10 rs10004195 loci genotype and the risk of Hp infection, but no correlation with Hp associated dis-eases.
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Objective Investigating the expression of TLR3 and TLR7 mRNA in peripheral blood mononuclear cells (PBMC) of patients with chronic hepatitis C infection (CHC),to explore the effects of gender and age on Toll-like receptor expres-sion.Methods Peripheral blood was collected from 1 1 5 patients of chronic hepatitis C infection,1 1 3 healthy individuals.Ex-pression levels of TLR3 and TLR7 mRNA were detected by real-time quantitative PCR.Results The expression of TLR3 and TLR7 mRNA were significant difference between patients with CHC infection and healthy individuals (t=38.73,6.16, P0.05).The HCV-RNA load of premenopausal females was significant lower than young males and old females (t=3.49,2.51,P<0.05),the load of old males was lower than old females (t=2.35,P<0.05),however, there was no significant differences between old males and old females (t=1.20,P<0.05).Conclusion Gender and age dis-crepancies have a relationship with the expression of Toll-like receptors of patients with CHC infection,which may provide a theoretical basis and a new method for CHC.
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Objective To analyze the association of the single nucleotide polymorphism (SNP) of Toll-like receptor 7 (TLR7) and Toll-like receptor 9 (TLR9) with chronic hepatitis C virus (HCV)infection.Methods A total of 150 patients with chronic hepatitis C (CHC) admitted to Renmin Hospital of Wuhan University from January 2011 to May 2012 and 168 healthy controls were enrolled in the study.The genotypes of TLR7 IVS2-151 (rs179009) were detected by Sanger sequencing,and the genotypes of TLR9 T-1486C (rs187084) were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).SPSS 15.0 was used for statistical analysis,and goodness-of-fit test for HardyWeinberg equilibrium was also performed.Results The frequency of TLR7 IVS2-151G was higher in malepatients with CHC than that in male controls (41.4％ vs.21.6％,x2 =7.250,P =0.007,OR =0.389,95％ CI:0.194-0.781) ; however the female CHC patients had a higher frequency of TLR7 IVS2-151A than the female controls (76.9％ vs.63.1％,x2 =7.202,P =0.007,OR =1.942,95％ CI:1.192-3.164).No significant difference in the distribution of TLR9 T-1486C (rs187084) gene SNP was observed betweenCHC and control groups (P ＞0.05).Conclusion TLR7 IVS2-151 (rs179009) is correlated with HCV infection,which may be involved in the pathogenesis of CHC.
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Objective To explore the effect of hepatitis B virus(HBV) on the expression of apolipoprotein A1 (ApoA1) and its regulatory mechanism.Methods RT-PCR and Western blot were used to measure the expression of ApoA1 in HepG2 and HepG2.2.15 cells,serum ApoA1 and high density lipoprotein cholesterol(HDL-C) levels in patients with HBV infection and in healthy individuals were measured by biochemical analyzer,statistical difference was analyzed by SPSS13.0,HepG2 cells was co-transfected with ApoA1 promoter containing the luciferase gene and HBV infectious clone pHBV1.3,luciferase activity was measured,expression of ApoA1 in HepG2 cells was measured by RT-PCR and Western blot after transfected with pHBV1.3.Results Expression of ApoA1 mRNA and protein was lower in HepG2.2.15 cells than in HepG2 cells,serum ApoA1 and HDL-C levels were much lower in HBV patients as compared to healthy individuals( P＜0.05 ),HBV represses ApoA1 gene promoter activity,ApoA1 mRNA and protein expression in HepG2 cells.Conclusion HBV can inhibit the expression of ApoA1 bothin vivo and in vitro.
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Objective To explore the effect of hepatitis B virus(HBV) on the expression of apolipoprotein B(ApoB) and its regulatory mechanism. Methods mRNA and protein expression of ApoB in HepG2 and HepG2.2.15 cells was measured by RT-PCR and Western blot, serum ApoB levels in patients with HBV infection and in healthy individuals were measured by biochemical analyzer Olympus 5400, the expression of ApoB difference among healthy individuals, patients with chronic hepatitis B, liver cirrhosis, and hepatocellular carcinoma were analyzed, HBV infectious clone pHBV1.3 was tranfected into HepG2 cells,and expression of ApoB and microsomal triglyceride transfer protein(MTP) was measured by RT-PCR and Western blot. Results Expression of ApoB mRNA and protein was lower in HepG2.2.15 cells than in HepG2 cells, serum apoB levels was much lower in patients with chronic hepatitis B and liver cirrhosis as compared to healthy individuals( P ＜0.05 ), HBV could inhibit the expression of ApoB and MTP at mRNA and protein levels. Conclusion HBV may downregulate the synthesis and secretion of ApoB via inhibits the expression of MTP.
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Objective To explore the regulatory effect of LPS on the expression of interleukin 27 (IL-27),and uncover the relationships among cyclooxyenase-2(COX-2),prostaglandin E2(PGE2) and IL-27.Methods THP-1 cells were stimulated with different doses of LPS,IL-27 and PGE2 levels in the supernatants were measured by enzyme-linked immunosorbent assay(ELISA) at different time,expression of COX-2 was measured by RT-PCR and Western blot.Results IL-27 can be induced by LPS in THP-1 cells in a time and dose dependent fashion,and IL-27 induces COX-2 mRNA expression,COX-2 protein production,and PGE2 release in a time and dose fashion.Conclusion LPS can stimulate the relase of IL-27 at cell level,and IL-27 can induce the expression of COX-2 and production of PGE2.
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OBJECTIVE To investigate antibiotic resistance in nosocomial infections with Acinetobacter baumannii from lower respiratory tract in the intensive care unit(ICU) and guide clinically rational use of drug.METHODS Antibiotics susceptibility tests in 109 clinical isolates of A.baumannii from lower respiratiory tract were performed by K-B disk diffusion method.The results were judged according to CLSI 2007.As controls,standard strain was used simultaneously.RESULTS The sensitivity rate to cefoperazone/sulbactam was the highest,arriving at 77.1% and followed by imipenem(57.8%) and meropenem(54.9%).The other antibiotics were below 50.0%,generally.The serious cross-resistance phenomenon was in present.Pan-resistant strains had been detected.CONCLUSIONS The drug-resistant status in nosocomial infections with A.baumannii from lower respiratory tract in ICU is very serious.We should continue to strengthen the monitoring of the antibiotic resistance and prevent the outbreak epidemics of drug resistant strains in ICU.