BACKGROUND IFIH1 is a protein-coding gene.Disorders associated with IFIH1 include Aicardi-Goutières syndrome(AGS)type 7 and Singleton-Merten syndrome type 1.Related pathways include RIG-I/MDA5-mediated induction ...BACKGROUND IFIH1 is a protein-coding gene.Disorders associated with IFIH1 include Aicardi-Goutières syndrome(AGS)type 7 and Singleton-Merten syndrome type 1.Related pathways include RIG-I/MDA5-mediated induction of the interferon(IFN)-α/βpathway and the innate immune system.AGS type 7 is an autosomal dominant inflammatory disorder characterized by severe neurological impairment.In infancy,most patients present with psychomotor retardation,axial hypotonia,spasticity,and brain imaging changes Laboratory assessments showed increased IFN-αactivity with upregulation of IFN signaling and IFN-stimulated gene expression.Some patients develop normally in the early stage,and then have episodic neurological deficits.CASE SUMMARY The 5-year-old girl presented with postpartum height and weight growth retardation,language retardation,brain atrophy,convulsions,and growth hormone deficiency.DNA samples were obtained from peripheral blood from the child and her parents for whole-exome sequencing and test of genome-wide copy number variation.Heterozygous mutations in the IFIH1 gene were found.Physical examination at admission found that language development was delayed,the reaction to name calling was average,there was no communication with people,but there was eye contact,no social smile,and no autonomous language.However,the child had rich gesture language and body language,could understand instructions,had bad temper.When she wants to achieve something,she starts crying or shouting.Cardiopulmonary examination showed no obvious abnormality,and abdominal examination was normal.Bilateral muscle strength and muscle tone were symmetrical and slightly decreased.Physiological reflexes exist,but pathological reflexes were not elicited.CONCLUSION We reported the clinical characteristics of a Chinese child with a clinical diagnosis of AGS type 7,which expanded the mutational spectrum of the IFIH1 gene.展开更多
BACKGROUND Spinocerebellar ataxia recessive type 7(SCAR7)is a rare clinical manifestation beginning in childhood or adolescence.SCAR7 is caused by tripeptidyl peptidase 1(TPP1)gene mutations,and presents with cerebell...BACKGROUND Spinocerebellar ataxia recessive type 7(SCAR7)is a rare clinical manifestation beginning in childhood or adolescence.SCAR7 is caused by tripeptidyl peptidase 1(TPP1)gene mutations,and presents with cerebellar ataxia,pyramidal signs,neurocognitive impairment,deep paresthesia,and cerebellar atrophy.CASE SUMMARY Here,we describe a 25-year-old female patient in China who presented with increasing difficulty walking,falling easily,shaking limbs,instability holding items,slurred speech,coughing when drinking,palpitations,and frequent hunger and overeating.Magnetic resonance imaging showed cerebellar atrophy.Whole exome sequencing detected two compound heterozygous mutations in the TPP1 gene:c.1468G>A p.Glu490Lys and c.1417G>A p.Gly473Arg.Considering the patient’s clinical presentation and genetic test results,we hypothesized that complex heterozygous mutations cause TPP1 enzyme deficiency,which may lead to SCAR7.CONCLUSION We report the first case of SCAR7 from China.We also identify novel compound heterozygous mutations in the TPP1 gene associated with SCAR7,expanding the range of known disease-causing mutations for SCAR7.展开更多
17β-hydroxysteroid dehydrogenase(17β-HSD)type 1 is known as a critical target to block the final step of estrogen production in estrogen-dependent breast cancer.Recent confirmation of the role of dyhydroxytestostero...17β-hydroxysteroid dehydrogenase(17β-HSD)type 1 is known as a critical target to block the final step of estrogen production in estrogen-dependent breast cancer.Recent confirmation of the role of dyhydroxytestosterone(DHT)in counteracting estrogeninduced cell growth prompted us to study the reductive 17β-HSD type 7(17β-HSD7),which activates estrone while markedly inactivatingDHT.The role ofDHTin breast cancer cell proliferation isdemonstratedby its independent suppression of cell growthin the presence of a physiological concentration of estradiol(E2).Moreover,an integral analysis of a large number of clinical samples in Oncomine datasets demonstrated the overexpression of 17β-HSD7 in breast carcinoma.Inhibition of 17β-HSD7 in breast cancer cells resulted in a lower level of E2 and a higher level of DHT,successively induced regulation of cyclinD1,p21,Bcl-2,and Bik,consequently arrested cell cycle in the G0/G1 phase,and triggered apoptosis and auto-downregulation feedback of the enzyme.Such inhibition led to significant shrinkage of xenograft tumors with decreased cancer cell density and reduced 17β-HSD7 expression.Decreased plasma E2 and elevated plasma DHT levels were also found.Thus,the dual functional 17β-HSD7 is proposed as a novel target for estrogen-dependent breast cancer by regulating the balance of E2 andDHT.Thisdemonstrates aconceptual advance on the general belief that the major role of this enzyme is in cholesterol metabolism.展开更多
To investigate the molecular epidemiology and genetic variation of human adenovirus type 7(HAdV-7)in children with acute respiratory infections(ARI)in China.HAdV-7-positive respiratory samples collected from children ...To investigate the molecular epidemiology and genetic variation of human adenovirus type 7(HAdV-7)in children with acute respiratory infections(ARI)in China.HAdV-7-positive respiratory samples collected from children with ARI in Beijing,Shijiazhuang,Wenzhou and Guangzhou from 2014–2018 were selected for gene amplification and sequence analysis.Fifty-seven HAdV-7 clinical strains with hexon,penton base and fiber gene sequences were obtained.Meanwhile17 strains were selected randomly from different cities for whole genome sequencing.Phylogenetic and variation analyses were performed based on the obtained sequences,HAdV-7 prototype strain Gomen(AY594255),vaccine strains(AY495969 and AY594256)and representative sequences of strains.The phylogenetic trees constructed based on whole genome sequences,major capsid protein genes(hexon,penton base and fiber)and the early genes(E1,E2,E3 and E4)were not completely consistent.The HAdV-7 strains obtained in this study always clustered with most of the circulating strains worldwide from the 1980 s to the present.Compared with the HAdV-7 prototype strain Gomen(AY594255),some amino acid mutations in loop1 and loop2 of hexon and the RGD loop region of the penton base gene were observed.Recombination analysis showed that partial regions of 55 k Da protein and 100 kDa hexon-assembly associated protein genes among all HAdV-7 strains in this study were from HAdV-16 and HAdV-3,respectively.Our study demonstrated the molecular evolution characteristics of HAdV-7 strains circulating in China and provided basic reference data for the prevention,control and vaccine development of HAdV-7.展开更多
Background Spinocerebellar ataxia type 7 （SCA7） is known as an autosomal dominant cerebellar ataxia; patients with genetically confirmed diagnoses of SCA7 have increased rapidly in recent years.However, SCA7 is a ra...Background Spinocerebellar ataxia type 7 （SCA7） is known as an autosomal dominant cerebellar ataxia; patients with genetically confirmed diagnoses of SCA7 have increased rapidly in recent years.However, SCA7 is a rare subtype of SCA, and most data available about SCA7 are those of white people.The aim of the present study was to systematically review the prevalence and clinical and genetic aspects of SCA7 patients in East Asian population.Methods A search for publications on SCA7 was performed by using the ＂PubMed＂ database with the published language limited in English.Publications mainly focusing on the prevalence of SCA7 in patients with SCA and the clinical and genetic features of SCA7 patients were fully reviewed and analyzed.Results The prevalence of SCA7 in SCA patients ranged from 0 to 7.7%, which was similar to those reported previously.The clinical manifestations were typically present at the 30＇s of its victims （median, 29 years; interquartile range （IQR）,19.5-36.5 years）, and the symptoms appeared 15 years （（15.17±4.22） years） earlier on average in the offspring than in the parents.Gait ataxia and visual impairment were both found in all patients of whom the clinical features were described.Mutant SCA7 alleles contained 40-100 CAG repeats, with a median of 47 repeats （IQR, 44.5-50.0）; and the offspring had 13 more repeats on average compared with their parents （12.62±19.03）.A strong negative correlation was found between CAG repeat size and the onset age of patients （r=-0.739, P=0.000）.In addition, no significant difference was found in CAG repeat sizes between patients with visual impairment as the initial symptom and those with gait disturbance as their initial symptom （P=0.476）.Conclusions The prevalence of SCA7 in SCA patients, the age at onset and CAG repeats of SCA7 patients in East Asia are consistent with those of white people.However, larger population study is needed to assess the correlation between the CAG repeat size and initial symptoms of SCA7 patients in East Asia.展开更多
BACKGROUND Liver resection is an effective treatment for benign and malignant liver tumors.However,a method for preoperative evaluation of hepatic reserve has not yet been established.Previously reported assessments o...BACKGROUND Liver resection is an effective treatment for benign and malignant liver tumors.However,a method for preoperative evaluation of hepatic reserve has not yet been established.Previously reported assessments of preoperative hepatic reserve focused only on liver failure in the early postoperative period and did not consider the long-term recovery of hepatic reserve.When determining eligibility for hepatectomy,the underlying pathophysiology needs to be considered to determine if the functional hepatic reserve can withstand both surgery and any postoperative therapy.AIM To identify pre-hepatectomy factors associated with both early postoperative liver failure and long-term postoperative liver function recovery.METHODS This study was a retrospective cohort study.We retrospectively investigated 215 patients who underwent hepatectomy at our hospital between May 2013 and December 2016.Early post-hepatectomy liver failure(PHLF)was defined using the International Study Group of Liver Surgery’s definition of PHLF.Long-term postoperative recovery of liver function was defined as the time taken for serum total bilirubin and albumin levels to return to levels of<2 mg/dL and>2.8 g/dL,respectively,and the time taken for Child-Pugh score to return to Child-Pugh class A.RESULTS Preoperative type IV collagen 7S was identified as a significant independent factor associated with both PHLF and postoperative long-term recovery of liver function.Further analysis revealed that the time taken for the recovery of Child-Pugh scores and serum total bilirubin and albumin levels was significantly shorter in patients with type IV collagen 7S≤6 ng/mL than in those with type IV collagen 7S>6 ng/mL.In additional analyses,similar results were observed in patients without chronic viral hepatitis associated with fibrosis.CONCLUSION Preoperative type IV collagen 7S is a preoperative predictor of PHLF and longterm postoperative liver function recovery.It can also be used in patients without chronic hepatitis virus.展开更多
BACKGROUND Non-alcoholic fatty liver disease(NAFLD)is the most common cause of chronic liver disease and affects approximately 25%of the general global adult population.The prognosis of NAFLD patients with advanced li...BACKGROUND Non-alcoholic fatty liver disease(NAFLD)is the most common cause of chronic liver disease and affects approximately 25%of the general global adult population.The prognosis of NAFLD patients with advanced liver fibrosis is known to be poor.It is difficult to assess disease progression in all patients with NAFLD;thus,it is necessary to identify patients who will show poor prognosis.AIM To investigate the efficacy of non-invasive biomarkers for predicting disease progression in patients with NAFLD.METHODS We investigated biomarkers associated with mortality in patients with NAFLD who visited the Kawasaki Medical School General Medical Center from 1996 to 2018 and underwent liver biopsy and had been followed-up for>1 year.Cumulative overall mortality and liver-related events during follow-up were calculated using the Kaplan-Meier analysis and compared using log-rank testing.We calculated the odds ratio and performed receiver operating characteristic curve analysis with logistic regression analysis to determine the optimal cut-off value with the highest prognostic ability.RESULTS We enrolled 489 patients who were followed-up for a period of 1-22.2 years.In total,13 patients died(2.7%of total patients enrolled);7 patients died due to liverrelated causes.Poor prognosis was associated with liver fibrosis on histological examination but not with inflammation or steatosis.Blood biomarkers associated with mortality were platelet counts,albumin levels,and type IV collagen 7S levels.The optimal cutoff index for predicting total mortality was a platelet count of 15×10^(4)/μL,albumin level of 3.5 g/dL,and type IV collagen 7S level of 5 mg/dL.In particular,only one-factor patients with NAFLD presenting with platelet counts≤15×10^(4)/μL,albumin levels≤3.5 g/dL,or type IV collagen 7S≥5 mg/dL showed 5-year,10-year,and 15-year survival rates of 99.7%,98.3%,and 94%,respectively.However,patients with two factors had lower 5-year and 10-year survival rates of 98%and 43%,respectively.Similarly,patients with all three factors showed the lowest 5-year and 10-year survival rates of 53%and 26%,respectively.CONCLUSION A combination of the three non-invasive biomarkers is a useful predictor of NAFLD prognosis and can help identify patients with NAFLD who are at a high risk of all-cause mortality.展开更多
Objective： The aim of this study was to study the effect of human papilloma virus （HPV） type 16 E7 protein ex- pression on growth of RMA cells in vitro and in vivo. Methods： The recombination vector pcDNA3.1-E7 ca...Objective： The aim of this study was to study the effect of human papilloma virus （HPV） type 16 E7 protein ex- pression on growth of RMA cells in vitro and in vivo. Methods： The recombination vector pcDNA3.1-E7 carrying wild type HPV 16 E7 was identified by sequencing. The recombination vector pcDNA3.1-E7 was transfected into mouse lymphadenoma cell line RMA by liposome, and the monoclonal cells transfected stably were obtained by antibiotics G418 sieving and limiting dilution assay. RT-PCR method was used to detect the expression of HPV 16 E7 mRNA in RMA-E7 cells. The growth of RMA cells and RMA-E7 cells cultured in vitro was tested by Cell Count Kit-8. RMA-E7 cells and RMA cells were subcutaneously inoculated in syngeneic mice respectively, the tumor size was measured by sliding caliper twice a week, and the E7 protein expression in tumor tissue of mice was detected by Western blot after tumor formation. The kinetics of cytolytic activity of E7 specific T cells in tumor-bearing mice was measured by LDH kit. Results： Sequencing of recombination vector showed the target gene which was inserted into the recombinant was correct, and RMA-E7 cells expressing E7 protein stably were obtained by limited dilution assay. There were no obvious differences in morphous and growth velocity between RMA cells and RMA-E7 cells in vitro. RMA-E7 cells grew in syngeneic mice were significantly slower than RMA cells. The E7 protein was ex- pressed stronger in RMA-E7 cells in vivo than in vitro. The cytolytic ability of ET-specific CTL was activated at the early stage, reached the maximum at the middle stage, and lost at the end stage. RMA-E7 cells isolated from the tumor-bearing mice were more resistant to E7-specific CTL killing than RMA-E7 cells cultured in vitro. Conclusion： The E7 protein expression has no obvious influence on growth of RMA-E7 cells in vitro, and can suppress growth of RMA-E7 cells in vivo. The activity curve of E7 specific CTL approximately presents ＂bell＂ shape. The RMA-E7 cells grew in vivo had a high expression levels of E7 protein, and more resistant to E7-specific CTL killing than those cultured in vitro. The E7 protein expression in vivo not only initiates immune activation, but also induces immune tolerance.展开更多
文摘BACKGROUND IFIH1 is a protein-coding gene.Disorders associated with IFIH1 include Aicardi-Goutières syndrome(AGS)type 7 and Singleton-Merten syndrome type 1.Related pathways include RIG-I/MDA5-mediated induction of the interferon(IFN)-α/βpathway and the innate immune system.AGS type 7 is an autosomal dominant inflammatory disorder characterized by severe neurological impairment.In infancy,most patients present with psychomotor retardation,axial hypotonia,spasticity,and brain imaging changes Laboratory assessments showed increased IFN-αactivity with upregulation of IFN signaling and IFN-stimulated gene expression.Some patients develop normally in the early stage,and then have episodic neurological deficits.CASE SUMMARY The 5-year-old girl presented with postpartum height and weight growth retardation,language retardation,brain atrophy,convulsions,and growth hormone deficiency.DNA samples were obtained from peripheral blood from the child and her parents for whole-exome sequencing and test of genome-wide copy number variation.Heterozygous mutations in the IFIH1 gene were found.Physical examination at admission found that language development was delayed,the reaction to name calling was average,there was no communication with people,but there was eye contact,no social smile,and no autonomous language.However,the child had rich gesture language and body language,could understand instructions,had bad temper.When she wants to achieve something,she starts crying or shouting.Cardiopulmonary examination showed no obvious abnormality,and abdominal examination was normal.Bilateral muscle strength and muscle tone were symmetrical and slightly decreased.Physiological reflexes exist,but pathological reflexes were not elicited.CONCLUSION We reported the clinical characteristics of a Chinese child with a clinical diagnosis of AGS type 7,which expanded the mutational spectrum of the IFIH1 gene.
基金Supported by Postdoctoral program of the Affiliated Hospital of Jining Medical University,No.JYFY303573Health Commission of Shandong Province,No.202006010928+1 种基金Academician Lin He New Medicine in Jining Medical University,No.JYHL2018FMS05Affiliated Hospital of Jining Medical University,No.2018-BS-004.
文摘BACKGROUND Spinocerebellar ataxia recessive type 7(SCAR7)is a rare clinical manifestation beginning in childhood or adolescence.SCAR7 is caused by tripeptidyl peptidase 1(TPP1)gene mutations,and presents with cerebellar ataxia,pyramidal signs,neurocognitive impairment,deep paresthesia,and cerebellar atrophy.CASE SUMMARY Here,we describe a 25-year-old female patient in China who presented with increasing difficulty walking,falling easily,shaking limbs,instability holding items,slurred speech,coughing when drinking,palpitations,and frequent hunger and overeating.Magnetic resonance imaging showed cerebellar atrophy.Whole exome sequencing detected two compound heterozygous mutations in the TPP1 gene:c.1468G>A p.Glu490Lys and c.1417G>A p.Gly473Arg.Considering the patient’s clinical presentation and genetic test results,we hypothesized that complex heterozygous mutations cause TPP1 enzyme deficiency,which may lead to SCAR7.CONCLUSION We report the first case of SCAR7 from China.We also identify novel compound heterozygous mutations in the TPP1 gene associated with SCAR7,expanding the range of known disease-causing mutations for SCAR7.
基金supported by operating grants from Canadian Institutes of Health Research(CIHR,MOP 97917 to S.-X.L.,D.P.,and C.J.D.MOP 89851 to S.-X.L.and D.P.)China Scholarship Council(PhD Fellowship,#2010621032 to X.Q.W.).
文摘17β-hydroxysteroid dehydrogenase(17β-HSD)type 1 is known as a critical target to block the final step of estrogen production in estrogen-dependent breast cancer.Recent confirmation of the role of dyhydroxytestosterone(DHT)in counteracting estrogeninduced cell growth prompted us to study the reductive 17β-HSD type 7(17β-HSD7),which activates estrone while markedly inactivatingDHT.The role ofDHTin breast cancer cell proliferation isdemonstratedby its independent suppression of cell growthin the presence of a physiological concentration of estradiol(E2).Moreover,an integral analysis of a large number of clinical samples in Oncomine datasets demonstrated the overexpression of 17β-HSD7 in breast carcinoma.Inhibition of 17β-HSD7 in breast cancer cells resulted in a lower level of E2 and a higher level of DHT,successively induced regulation of cyclinD1,p21,Bcl-2,and Bik,consequently arrested cell cycle in the G0/G1 phase,and triggered apoptosis and auto-downregulation feedback of the enzyme.Such inhibition led to significant shrinkage of xenograft tumors with decreased cancer cell density and reduced 17β-HSD7 expression.Decreased plasma E2 and elevated plasma DHT levels were also found.Thus,the dual functional 17β-HSD7 is proposed as a novel target for estrogen-dependent breast cancer by regulating the balance of E2 andDHT.Thisdemonstrates aconceptual advance on the general belief that the major role of this enzyme is in cholesterol metabolism.
基金funded by the Key Technology R&D Program of China(grant numbers2017ZX10103004-004,2017ZX10104001-005-010)National Natural Science Foundation of China(grant number 82072266)CAMS Innovation Fund for Medical Sciences(CIFMS),(grant number 2019-I2M-5-026)。
文摘To investigate the molecular epidemiology and genetic variation of human adenovirus type 7(HAdV-7)in children with acute respiratory infections(ARI)in China.HAdV-7-positive respiratory samples collected from children with ARI in Beijing,Shijiazhuang,Wenzhou and Guangzhou from 2014–2018 were selected for gene amplification and sequence analysis.Fifty-seven HAdV-7 clinical strains with hexon,penton base and fiber gene sequences were obtained.Meanwhile17 strains were selected randomly from different cities for whole genome sequencing.Phylogenetic and variation analyses were performed based on the obtained sequences,HAdV-7 prototype strain Gomen(AY594255),vaccine strains(AY495969 and AY594256)and representative sequences of strains.The phylogenetic trees constructed based on whole genome sequences,major capsid protein genes(hexon,penton base and fiber)and the early genes(E1,E2,E3 and E4)were not completely consistent.The HAdV-7 strains obtained in this study always clustered with most of the circulating strains worldwide from the 1980 s to the present.Compared with the HAdV-7 prototype strain Gomen(AY594255),some amino acid mutations in loop1 and loop2 of hexon and the RGD loop region of the penton base gene were observed.Recombination analysis showed that partial regions of 55 k Da protein and 100 kDa hexon-assembly associated protein genes among all HAdV-7 strains in this study were from HAdV-16 and HAdV-3,respectively.Our study demonstrated the molecular evolution characteristics of HAdV-7 strains circulating in China and provided basic reference data for the prevention,control and vaccine development of HAdV-7.
文摘Background Spinocerebellar ataxia type 7 （SCA7） is known as an autosomal dominant cerebellar ataxia; patients with genetically confirmed diagnoses of SCA7 have increased rapidly in recent years.However, SCA7 is a rare subtype of SCA, and most data available about SCA7 are those of white people.The aim of the present study was to systematically review the prevalence and clinical and genetic aspects of SCA7 patients in East Asian population.Methods A search for publications on SCA7 was performed by using the ＂PubMed＂ database with the published language limited in English.Publications mainly focusing on the prevalence of SCA7 in patients with SCA and the clinical and genetic features of SCA7 patients were fully reviewed and analyzed.Results The prevalence of SCA7 in SCA patients ranged from 0 to 7.7%, which was similar to those reported previously.The clinical manifestations were typically present at the 30＇s of its victims （median, 29 years; interquartile range （IQR）,19.5-36.5 years）, and the symptoms appeared 15 years （（15.17±4.22） years） earlier on average in the offspring than in the parents.Gait ataxia and visual impairment were both found in all patients of whom the clinical features were described.Mutant SCA7 alleles contained 40-100 CAG repeats, with a median of 47 repeats （IQR, 44.5-50.0）; and the offspring had 13 more repeats on average compared with their parents （12.62±19.03）.A strong negative correlation was found between CAG repeat size and the onset age of patients （r=-0.739, P=0.000）.In addition, no significant difference was found in CAG repeat sizes between patients with visual impairment as the initial symptom and those with gait disturbance as their initial symptom （P=0.476）.Conclusions The prevalence of SCA7 in SCA patients, the age at onset and CAG repeats of SCA7 patients in East Asia are consistent with those of white people.However, larger population study is needed to assess the correlation between the CAG repeat size and initial symptoms of SCA7 patients in East Asia.
文摘BACKGROUND Liver resection is an effective treatment for benign and malignant liver tumors.However,a method for preoperative evaluation of hepatic reserve has not yet been established.Previously reported assessments of preoperative hepatic reserve focused only on liver failure in the early postoperative period and did not consider the long-term recovery of hepatic reserve.When determining eligibility for hepatectomy,the underlying pathophysiology needs to be considered to determine if the functional hepatic reserve can withstand both surgery and any postoperative therapy.AIM To identify pre-hepatectomy factors associated with both early postoperative liver failure and long-term postoperative liver function recovery.METHODS This study was a retrospective cohort study.We retrospectively investigated 215 patients who underwent hepatectomy at our hospital between May 2013 and December 2016.Early post-hepatectomy liver failure(PHLF)was defined using the International Study Group of Liver Surgery’s definition of PHLF.Long-term postoperative recovery of liver function was defined as the time taken for serum total bilirubin and albumin levels to return to levels of<2 mg/dL and>2.8 g/dL,respectively,and the time taken for Child-Pugh score to return to Child-Pugh class A.RESULTS Preoperative type IV collagen 7S was identified as a significant independent factor associated with both PHLF and postoperative long-term recovery of liver function.Further analysis revealed that the time taken for the recovery of Child-Pugh scores and serum total bilirubin and albumin levels was significantly shorter in patients with type IV collagen 7S≤6 ng/mL than in those with type IV collagen 7S>6 ng/mL.In additional analyses,similar results were observed in patients without chronic viral hepatitis associated with fibrosis.CONCLUSION Preoperative type IV collagen 7S is a preoperative predictor of PHLF and longterm postoperative liver function recovery.It can also be used in patients without chronic hepatitis virus.
文摘BACKGROUND Non-alcoholic fatty liver disease(NAFLD)is the most common cause of chronic liver disease and affects approximately 25%of the general global adult population.The prognosis of NAFLD patients with advanced liver fibrosis is known to be poor.It is difficult to assess disease progression in all patients with NAFLD;thus,it is necessary to identify patients who will show poor prognosis.AIM To investigate the efficacy of non-invasive biomarkers for predicting disease progression in patients with NAFLD.METHODS We investigated biomarkers associated with mortality in patients with NAFLD who visited the Kawasaki Medical School General Medical Center from 1996 to 2018 and underwent liver biopsy and had been followed-up for>1 year.Cumulative overall mortality and liver-related events during follow-up were calculated using the Kaplan-Meier analysis and compared using log-rank testing.We calculated the odds ratio and performed receiver operating characteristic curve analysis with logistic regression analysis to determine the optimal cut-off value with the highest prognostic ability.RESULTS We enrolled 489 patients who were followed-up for a period of 1-22.2 years.In total,13 patients died(2.7%of total patients enrolled);7 patients died due to liverrelated causes.Poor prognosis was associated with liver fibrosis on histological examination but not with inflammation or steatosis.Blood biomarkers associated with mortality were platelet counts,albumin levels,and type IV collagen 7S levels.The optimal cutoff index for predicting total mortality was a platelet count of 15×10^(4)/μL,albumin level of 3.5 g/dL,and type IV collagen 7S level of 5 mg/dL.In particular,only one-factor patients with NAFLD presenting with platelet counts≤15×10^(4)/μL,albumin levels≤3.5 g/dL,or type IV collagen 7S≥5 mg/dL showed 5-year,10-year,and 15-year survival rates of 99.7%,98.3%,and 94%,respectively.However,patients with two factors had lower 5-year and 10-year survival rates of 98%and 43%,respectively.Similarly,patients with all three factors showed the lowest 5-year and 10-year survival rates of 53%and 26%,respectively.CONCLUSION A combination of the three non-invasive biomarkers is a useful predictor of NAFLD prognosis and can help identify patients with NAFLD who are at a high risk of all-cause mortality.
文摘Objective： The aim of this study was to study the effect of human papilloma virus （HPV） type 16 E7 protein ex- pression on growth of RMA cells in vitro and in vivo. Methods： The recombination vector pcDNA3.1-E7 carrying wild type HPV 16 E7 was identified by sequencing. The recombination vector pcDNA3.1-E7 was transfected into mouse lymphadenoma cell line RMA by liposome, and the monoclonal cells transfected stably were obtained by antibiotics G418 sieving and limiting dilution assay. RT-PCR method was used to detect the expression of HPV 16 E7 mRNA in RMA-E7 cells. The growth of RMA cells and RMA-E7 cells cultured in vitro was tested by Cell Count Kit-8. RMA-E7 cells and RMA cells were subcutaneously inoculated in syngeneic mice respectively, the tumor size was measured by sliding caliper twice a week, and the E7 protein expression in tumor tissue of mice was detected by Western blot after tumor formation. The kinetics of cytolytic activity of E7 specific T cells in tumor-bearing mice was measured by LDH kit. Results： Sequencing of recombination vector showed the target gene which was inserted into the recombinant was correct, and RMA-E7 cells expressing E7 protein stably were obtained by limited dilution assay. There were no obvious differences in morphous and growth velocity between RMA cells and RMA-E7 cells in vitro. RMA-E7 cells grew in syngeneic mice were significantly slower than RMA cells. The E7 protein was ex- pressed stronger in RMA-E7 cells in vivo than in vitro. The cytolytic ability of ET-specific CTL was activated at the early stage, reached the maximum at the middle stage, and lost at the end stage. RMA-E7 cells isolated from the tumor-bearing mice were more resistant to E7-specific CTL killing than RMA-E7 cells cultured in vitro. Conclusion： The E7 protein expression has no obvious influence on growth of RMA-E7 cells in vitro, and can suppress growth of RMA-E7 cells in vivo. The activity curve of E7 specific CTL approximately presents ＂bell＂ shape. The RMA-E7 cells grew in vivo had a high expression levels of E7 protein, and more resistant to E7-specific CTL killing than those cultured in vitro. The E7 protein expression in vivo not only initiates immune activation, but also induces immune tolerance.
