Background:Calcaneus is the largest bone of foot and the main load-bearing structure of heel.The incidence of simple calcaneal tubercle avulsion fracture is low,accounting for about 1%to 3%of all calcaneal fractures.B...Background:Calcaneus is the largest bone of foot and the main load-bearing structure of heel.The incidence of simple calcaneal tubercle avulsion fracture is low,accounting for about 1%to 3%of all calcaneal fractures.Beavis II fracture has large bone fracture,obvious displacement,obvious soft tissue irritation,and often leads to skin necrosis.It needs surgical treatment,reduction and fixation as soon as possible.Although open reduction and tension screw internal fixation is used for Beavis II calcaneal tubercle fracture,but the failure rate is more common.Methods:This study retrospectively analyzed the surgical treatment of calcaneal tubercle Beavis II fracture over 55 years old in our hospital from January 2013 to January 2019.The patients were treated with tension screw combined with locking plate,and followed up and analyzed.Results:12 patients in this group were followed up for 12 to 36 months(mean 20 months).After operation,the fracture healed smoothly in all patients,the healing time was 8 to 12 weeks(mean 10.7 weeks),and there were no complications such as poor incision healing,fracture displacement,internal fixation loosening,fracture and so on.When the patients were followed up 18 weeks after operation,the AOFAS score was 47 to 100,with an average of 91.1,of which 8 cases were excellent,3 good and 1 poor,with an excellent and good rate of 91.7%.Conclusion:Our hospital has been treated with tension screw combined with locking plate,fixed firmly,can early functional exercise,achieved good results.展开更多
Objective:To investigate the effects of hepatitis B virus(HBV)X protein(HBx)on the expression of tumor necrosis factor-α(TNF-α)in glomerular mesangial cells(GMCs)and the underlying intracellular signal pathways.Meth...Objective:To investigate the effects of hepatitis B virus(HBV)X protein(HBx)on the expression of tumor necrosis factor-α(TNF-α)in glomerular mesangial cells(GMCs)and the underlying intracellular signal pathways.Methods:The plasmid pCI-neo-X that carries the X gene of hepatitis B virus was transfected into cultured GMCs.HBx expression in the transfected GMCs was assessed by Western-blot.TNF-αprotein and mRNA were assessed by ELISA and semi-quantitative RT-PCR,respectively.Three kinase inhibitors-U0126,an inhibitor of extracellular signal-regulated kinases(ERKs);lactacvstin,an inhibitor of nuclear factor-κB(NF-κB);and SB203580,a selective inhibitor of p38 MAP kinase(p38 MAPK)were used to determine which intracellular signal pathways may underlie the action of HBx on TNF-αexpression in transfected GMCs.Results:A significant increase in HBx expression in pCI-neo-X transfected GMCs was detected at 36 h and 48 h,which was not affected by any of those kinase inhibitors mentioned above.A similar increase in the expression of both TNF-αprotein and mRNA was also observed at 36 h and 48 h,which was significantly decreased in the presence of U0126 or lactacytin,but not SB203580.Conclusions:HBx upregulates TNF-αexpression in cultured GMCs,possibly through ERKs and NF-κB pathway,but not p38 MAPK pathway.展开更多
AIM: To study the effect of the transfected Twist gene on invasion and metastasis of gastric carcinoma cells and the possible mechanisms involved. METHODS: Human gastric carcinoma MKN28 cells were stably transfected w...AIM: To study the effect of the transfected Twist gene on invasion and metastasis of gastric carcinoma cells and the possible mechanisms involved. METHODS: Human gastric carcinoma MKN28 cells were stably transfected with Twist sense plasmid, and MKN45 cells were stably transfected with Twist antisense plasmid using the lipofectamine transfection technique. RT-PCR, Western blotting, EMSA, gelatin zymography assay, and in vitro invasion and migration assays were performed. Nude mice metastasis models were established by the abdominal cavity transfer method. RESULTS: Cell models (TwistS-MKN28) that steadily expressed high Twist protein were obtained. Compared with MKN28 and pcDNA3-MKN28 cells, adherence, migration and invasion ability of TwistS -MKN28 cells were clearly raised. The number of cancer nodules was increased significantly in the abdominal cavity and liver of nude mice inoculated with TwistS-MKN28 cells. Overexpression of Twist in MKN28 cells increased Tcf-4/ Lef DNA binding activity, and promoted expression of Tcf-4’s downstream target genes cyclin D1 and MMP-2. However, suppression of Twist (TwistAS-MKN45) inhibited MKN45 cell invasion and the expression of cyclin D1 was reduced. The activity of MMP-2 was also decreased. CONCLUSION: These results indicate that Twist promotes gastric cancer cell migration, invasion and metastasis, and Twist may play an important role in Wnt/ Tcf-4 signaling.展开更多
Vesicoureteral reflux(VUR)is one of the most common urinary tract anomalies in children and causes renal damage and studies focusing on the effect of VUR on renal function are rare.We recruited 35 primary VUR patients...Vesicoureteral reflux(VUR)is one of the most common urinary tract anomalies in children and causes renal damage and studies focusing on the effect of VUR on renal function are rare.We recruited 35 primary VUR patients with recurrent urinary tract infection(UTI)and 10 non-VUR patients with recurrent UTI.Contrast-enhanced voiding urosonography(ceVUS)was performed for VUR grading,and renal dynamic imaging was used for evaluating glomerular filtration rate(GFR,mL/min).Standardized GFR(sGFR),namely GFR/BSA(mL·min-1·m-2),was calculated based on the body surface area(BSA).Total sGFR(tsGFR,mL·min-1·m-2)was obtained from the sum of sGFR on the left and right sides of all the children.The risk of renal regurgitation was equal in the unilateral reflux group.The sGFR of children with grade Ⅳ(45.74±18.05mL·min-1·m-2)and grade V(49.67±23.63mL·min-1·m-2)reflux was significantly lower than that in children with grade Ⅱ(77.69±22.21 mL·min-1·m-2).The renal function compensation of contralateral non-reflux kidney increased in unilateral reflux group,which was higher than that in the control group and level Ⅱ,Ⅳ and Ⅴ of reflux group respectively.In VUR group of the same grade,sGFR decreased with the age at diagnosis.In unilateral grade V refux group,the tsGFR was lower than that in the unilateral grade I reflux group(133.51±48.21 vs.186.87+53.49mL·min-1·m-2).The patients with VUR of unilateral grade Ⅱ were significantly older than those with VUR of unilateral grades Ⅱ and Ⅳ.This study indicates that severe VUR is significantly associated with decreased renal function.Therefore,VUR should be diagnosed early and managed individually.展开更多
PKHD1 mutations are generally considered to cause autosomal recessive polycystic kidney disease(ARPKD).ARPKD is a rare disorder and one o f the most severe conditions leading to end-stage renal disease in childhood.Wi...PKHD1 mutations are generally considered to cause autosomal recessive polycystic kidney disease(ARPKD).ARPKD is a rare disorder and one o f the most severe conditions leading to end-stage renal disease in childhood.With the biallelic deletion mutation,patients have difficulty in surviving the perinatal period,resulting in perinatal or neonatal death.This study retrospectively analyzed patient characteristics,imaging characteristics,laboratory examinations and family surveys from 7 Chinese children with different PKHD1 gene mutations diagnosed by high-throughput sequencing from January 2014 to February 2018.O f the 7 children,there were 3 males and 4 females.Eight missense mutations,two frameshift mutations,two deletion mutations,and two intronic slicing mutations were identified.Six of the mutations have not previously been identified.In the literature search,we identified a total of 29 Chinese children with PKHD1 mutations.The missense mutation c.2507T>C in exon 24 was found in one patient in our study,and five patients with liver fibrosis but normal renal function were reported in the literature.The missense mutation c.5935G>A in exon 37 was found in two patients in our study and three cases in the literature.Four patients had renal failure at an age as young as 1 year of those five patients with the missense mutation c.5935G>A in exon 37.It was concluded that:(1)Kidney length more than 2-3 SDs above the mean and early-onset hypertension might be associated with PKHDI-associated ARPICD;(2)The more enlarged the kidney size is,the lower the renal function is likely to be;(3)c.5935G>A may be a hot spot that leads to early renal failure in Chinese children with PKHD1 mutations;(4)c.2507T>C may be a hot-spot mutation associated with hepatic lesions in Chinese children with PKHD1.展开更多
Objective:Autosomal dominant polycystic kidney disease(ADPKD)is mainly caused by the pathogenic mutation of PKD1 or PKD2 gene and usually affects bilateral kidneys.Synonymous mutations are generally assumed to be neut...Objective:Autosomal dominant polycystic kidney disease(ADPKD)is mainly caused by the pathogenic mutation of PKD1 or PKD2 gene and usually affects bilateral kidneys.Synonymous mutations are generally assumed to be neutral as they do not alter amino acids.Herein,we described an extremely rare ADPKD child caused by a heterozygous synonymous mutation of PKD2 gene accompanied by massive proteinuria and congenital solitary kidney.Methods:Clinical characteristics of the patients were summarized.Whole-exome sequencing was performed to screen the disease-causing gene mutation,and reverse transcription polymerase chain reaction(RT-PCR)and Sanger sequencing were applied to analyze the impact of the identified mutation on gene transcription and splicing.Results:Polycystic changes were found in the solitary kidney of a girl initially presented with nephrotic-range proteinuria.Thereafter her mother and 2 other family members were diagnosed to be ADPKD.Whole-exome sequencing of the proband identified a heterozygous synonymous mutation(c.1716G>A,p.Lys572=)located in the splicing site of exon 7 in PKD2 gene,which was co-segregated with the PKD phenotype in the family.RT-PCR and direct sequencing of amplified products revealed that this heterozygous synonymous mutation led to exon7 skipping in PKD2 gene.Conclusion:We reported an extremely rare child case of ADPKD2 in combination with solitary kidney and nephrotic-range proteinuria,and firstly confirmed the pathogenicity of a heterozygous synonymous mutation(c.1716G>A)in PKD2 gene.The results indicate that synonymous mutations should not be excluded from disease-causing if they are located in splicing site of an exon.展开更多
Objective Ubiquitin-specific protease 4(USP4)facilitates the development of transforming growth factor-beta 1(TGF-β1)-induced epithelial-mesenchymal transition(EMT)in various cancer cells.Moreover,EMT of renal tubula...Objective Ubiquitin-specific protease 4(USP4)facilitates the development of transforming growth factor-beta 1(TGF-β1)-induced epithelial-mesenchymal transition(EMT)in various cancer cells.Moreover,EMT of renal tubular epithelial cells(RTECs)is required for the progression of renal interstitial fibrosis.However,the role of USP4 in EMT of RTECs remains unknown.The present study aimed to explore the effect of USP4 on the EMT of RTECs as well as the involved mechanism.Methods In established unilateral ureteral obstruction(UUO)rats and NRK-52E cells,immunohistochemistry and Western blot assays were performed.Results USP4 expression was increased significantly with obstruction time.In NRK-52E cells stimulated by TGF-β1,USP4 expression was increased in a time-dependent manner.In addition,USP4 silencing with specific siRNA indicated that USP4 protein was suppressed effectively.Meanwhile,USP4 siRNA treatment restored E-cadherin and weakened alpha smooth muscle actin(α-SMA)expression,indicating that USP4 may promote EMT.After treatment with USP4 siRNA and TGF-β1 for 24 h,the expression of TGF-β1 receptor type I(TβRI)was decreased.Conclusion USP4 promotes the EMT of RTECs through upregulating TβRI,thereby facilitating renal interstitial fibrosis.These findings may provide a potential target of USP4 in the treatment of renal fibrosis.展开更多
Background: Current knowledge about clinical and genetic risk factors for aspirin-induced gastric mucosal injury is not sufficient to prevent these gastric mucosal lesions. Methods: We recruited aspirin takers as th...Background: Current knowledge about clinical and genetic risk factors for aspirin-induced gastric mucosal injury is not sufficient to prevent these gastric mucosal lesions. Methods: We recruited aspirin takers as the exposed group and healthy volunteers as the control group. The exposed group was categorized into two subgroups such as subgroup A as gastric mucosal injury diagnosed by gastroscopy, including erosion, ulcer or bleeding of the esophagus, stomach, or duodenum; subgroup B as no injury of the gastric mucosa was detected by gastroscopy. Clinical information was collected, and 53 single nucleotide polymorphisms were evaluated. Results: Among 385 participants, 234 were in the aspirin-exposed group. According to gastroscopy, 82 belonged to subgroup A, 91 belonged to subgroup B, and gastroscopic results of 61 participants were not available. Using the Chi-square test and logistic regression, we found that peptic ulcer history (odds ratio [OR] = 5.924, 95% confidence intervals [C/]: 2.115-16.592), dual anti-platelet medication (OR = 3.443, 95% CI: 1.154-10.271 ), current Helicobacterpylori infection (OR = 2.242, 95% CI: 1.032-4.870), male gender (OR = 2.211, 95% CI: 1.027-4.760), GG genotype ofrs2243086 (OR = 4.516, 95% CI: I. 180-17.278), and AA genotype ofrs 1330344 (OR = 2.178, 95% CI: 1.016-4.669) were more frequent in subgroup A than subgroup B. In aspirin users who suffered from upper gastrointestinal bleeding, the frequency of the TT genotype ofrs2238631 and TT genotype ofrs2243100 was higher than in those without upper gastrointestinal bleeding. Conclusions: Peptic ulcer history, dual anti-platelet medication, tt. pylori current infection, and male gender were possible clinical risk factors for aspirin-induced gastric mucosal injury. GG genotype of rs2243086 and AA genotype of rs 1330344 were possible genetic risk factors. TT genotype ofrs2238631 and TT genotype ofrs2243100 may be risk factors for upper gastrointestinal bleeding in aspirin users.展开更多
Hepatitis B virus X protein(HBx),a 17-kd protein encoded by X gene of hepatitis B virus(HBV),has been shown to function as a transcriptional trans-activator of a variety of viral and cellular promoter/enhancer element...Hepatitis B virus X protein(HBx),a 17-kd protein encoded by X gene of hepatitis B virus(HBV),has been shown to function as a transcriptional trans-activator of a variety of viral and cellular promoter/enhancer elements.The aim of the study is to investigate the extracellular regulated protein kinases(ERKs)pathway of HBx on glomerular mesangial cell(GMC)proliferation and tumor necrosis factor-α(TNF-α)expression.The HBV X gene was amplified by polymerase chain reaction(PCR),inserted into the eukaryotic expression vector pCI-neo and confirmed by restriction endonuclease digestion and sequence analysis.PCI-neo containing HBV X gene(pCI-neo-X)was then transfected into cultured GMC line via liposome.GMC proliferation,TNF-αand its mRNA expression were compared in the condition of with or without U0126 in culture media.HBx,ERK1/2 and p-ERK1/2 expression in GMCs was assessed by Western blotting.TNF-αmRNA expression was assessed by semi-quantitative reverse transcription-PCR(RT-PCR).TNF-αlevel in supernatants was measured by ELISA.GMC proliferation was detected by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide(MTT)kit.The results showed that HBx expression was found in transfected GMCs and became prominent at 36th and 48th h after transfection whether with or without U0126 in culture media.TNF-αmRNA expression was significantly decreased in U0126 group compared with U0126-free group.TNF-αlevels in supernatants in PCI-neo-X transfection without U0126 group were(189.0�18.1)and(172.3�24.3)pg/mL at 36th and 48th h after transfec-tion,respectively.In contrast,TNF-αlevels in supernatants with U0126 were(65.6�11.6)and(84.0�24.6)pg/mL at 36th and 48th h,respectively.The TNF-αlevels in the latter groups were significantly lower than those in the former groups(P<0.05).GMCs proliferation was also lower in added U0126 group at 36th and 48th h after transfection.From above,we can conclude that HBx could induce GMC proliferation and increase TNF-αmRNA expression and its protein production.HBx upregulates TNF-αexpression and induces cell proliferation of GMC line partly through ERK1/2 signal transduction pathway.展开更多
Objective:To explore the value of transrectal ultrasonography(TRUS)for tumor node metastasis(TNM)restaging for patients with locally advanced rectal cancer after neoadjuvant chemoradiotherapy(neo-CRT).Methods:One hund...Objective:To explore the value of transrectal ultrasonography(TRUS)for tumor node metastasis(TNM)restaging for patients with locally advanced rectal cancer after neoadjuvant chemoradiotherapy(neo-CRT).Methods:One hundred and forty-nine patients with locally advanced rectal cancer(cT3-4 or cN+)who underwent TRUS after neo-CRT were retrospectively reviewed.TRUS restaging was compared with the results of post-operative pathological TNM findings.Results:After neo-CRT,the accuracy of TRUS for diagnosing T-staging was 30.9%,with 60.4%(90/149)of cases overestimated.The sensitivity of TRUS for T-staging(T0 vs T1 vs T2 vs T3 vs T4)were 16.3%,0%,12.5%,42.6%and 75.0%,respectively.The accuracy of TRUS for diagnosing N-staging after neo-CRT was 81.2%,with the sensitivities of N0 and N+were 93.3%and 31.0%,respectively.After neo-CRT,27.5%(41/149)of patients achieved pathologically complete response(pCR).The sensitivity,specificity,positive predictive value and negative predictive values of TRUS for pCR were 17.1%,99.1%,87.5%and 75.9%,respectively.Conclusions:TRUS can be applied for restaging T4 and N0,and has potential for screening out patients with pCR in those with locally advanced rectal cancer after neo-CRT,although some stages are overestimated for T-staging and its sensitivity for predicting pCR is low.展开更多
基金the Youth Science and Technology Project of Health Commission of Shanghai Pudong New Area(Project no.PW2020B-5)the Outstanding Clinical Discipline Project of Shanghai Pudong(Grant No.PWYgy2021-04)+4 种基金the Health Industry Clinical Research Project of Shanghai Health Commission(Project no.20224Y0393)the Young Medical Talents Training Program of Pudong Health Committee of Shanghai(Grant No.PWRq 2021-08)the Outstanding Leaders Training Program of Pudong Hospital affiliated to Fudan University(Grant No.LX202201)the Talents Training Program of Pudong Hospital affiliated to Fudan University(Project no.PX202001)the Scientific Research Foundation provided by Pudong Hospital affiliated to Fudan University(Project no.YJRCJJ201906).
文摘Background:Calcaneus is the largest bone of foot and the main load-bearing structure of heel.The incidence of simple calcaneal tubercle avulsion fracture is low,accounting for about 1%to 3%of all calcaneal fractures.Beavis II fracture has large bone fracture,obvious displacement,obvious soft tissue irritation,and often leads to skin necrosis.It needs surgical treatment,reduction and fixation as soon as possible.Although open reduction and tension screw internal fixation is used for Beavis II calcaneal tubercle fracture,but the failure rate is more common.Methods:This study retrospectively analyzed the surgical treatment of calcaneal tubercle Beavis II fracture over 55 years old in our hospital from January 2013 to January 2019.The patients were treated with tension screw combined with locking plate,and followed up and analyzed.Results:12 patients in this group were followed up for 12 to 36 months(mean 20 months).After operation,the fracture healed smoothly in all patients,the healing time was 8 to 12 weeks(mean 10.7 weeks),and there were no complications such as poor incision healing,fracture displacement,internal fixation loosening,fracture and so on.When the patients were followed up 18 weeks after operation,the AOFAS score was 47 to 100,with an average of 91.1,of which 8 cases were excellent,3 good and 1 poor,with an excellent and good rate of 91.7%.Conclusion:Our hospital has been treated with tension screw combined with locking plate,fixed firmly,can early functional exercise,achieved good results.
基金Supported by National Nature Science Foundation of China(GrantNo.30772360)Nature Science Foundation of Health Department of Hubei Province,China(No.JX4B48)Fund of Yangtze University for Doctor(No.2009001)
文摘Objective:To investigate the effects of hepatitis B virus(HBV)X protein(HBx)on the expression of tumor necrosis factor-α(TNF-α)in glomerular mesangial cells(GMCs)and the underlying intracellular signal pathways.Methods:The plasmid pCI-neo-X that carries the X gene of hepatitis B virus was transfected into cultured GMCs.HBx expression in the transfected GMCs was assessed by Western-blot.TNF-αprotein and mRNA were assessed by ELISA and semi-quantitative RT-PCR,respectively.Three kinase inhibitors-U0126,an inhibitor of extracellular signal-regulated kinases(ERKs);lactacvstin,an inhibitor of nuclear factor-κB(NF-κB);and SB203580,a selective inhibitor of p38 MAP kinase(p38 MAPK)were used to determine which intracellular signal pathways may underlie the action of HBx on TNF-αexpression in transfected GMCs.Results:A significant increase in HBx expression in pCI-neo-X transfected GMCs was detected at 36 h and 48 h,which was not affected by any of those kinase inhibitors mentioned above.A similar increase in the expression of both TNF-αprotein and mRNA was also observed at 36 h and 48 h,which was significantly decreased in the presence of U0126 or lactacytin,but not SB203580.Conclusions:HBx upregulates TNF-αexpression in cultured GMCs,possibly through ERKs and NF-κB pathway,but not p38 MAPK pathway.
文摘AIM: To study the effect of the transfected Twist gene on invasion and metastasis of gastric carcinoma cells and the possible mechanisms involved. METHODS: Human gastric carcinoma MKN28 cells were stably transfected with Twist sense plasmid, and MKN45 cells were stably transfected with Twist antisense plasmid using the lipofectamine transfection technique. RT-PCR, Western blotting, EMSA, gelatin zymography assay, and in vitro invasion and migration assays were performed. Nude mice metastasis models were established by the abdominal cavity transfer method. RESULTS: Cell models (TwistS-MKN28) that steadily expressed high Twist protein were obtained. Compared with MKN28 and pcDNA3-MKN28 cells, adherence, migration and invasion ability of TwistS -MKN28 cells were clearly raised. The number of cancer nodules was increased significantly in the abdominal cavity and liver of nude mice inoculated with TwistS-MKN28 cells. Overexpression of Twist in MKN28 cells increased Tcf-4/ Lef DNA binding activity, and promoted expression of Tcf-4’s downstream target genes cyclin D1 and MMP-2. However, suppression of Twist (TwistAS-MKN45) inhibited MKN45 cell invasion and the expression of cyclin D1 was reduced. The activity of MMP-2 was also decreased. CONCLUSION: These results indicate that Twist promotes gastric cancer cell migration, invasion and metastasis, and Twist may play an important role in Wnt/ Tcf-4 signaling.
基金This work was supported by the National Natural Science Foundation of China(No.11602121)the Program for Scientific Research Innovation Team in Colleges and Universities of Ji’nan(No.2018GXRC006).
基金This project was supported by the National Natural Science Foundation of China(No.81873596).
文摘Vesicoureteral reflux(VUR)is one of the most common urinary tract anomalies in children and causes renal damage and studies focusing on the effect of VUR on renal function are rare.We recruited 35 primary VUR patients with recurrent urinary tract infection(UTI)and 10 non-VUR patients with recurrent UTI.Contrast-enhanced voiding urosonography(ceVUS)was performed for VUR grading,and renal dynamic imaging was used for evaluating glomerular filtration rate(GFR,mL/min).Standardized GFR(sGFR),namely GFR/BSA(mL·min-1·m-2),was calculated based on the body surface area(BSA).Total sGFR(tsGFR,mL·min-1·m-2)was obtained from the sum of sGFR on the left and right sides of all the children.The risk of renal regurgitation was equal in the unilateral reflux group.The sGFR of children with grade Ⅳ(45.74±18.05mL·min-1·m-2)and grade V(49.67±23.63mL·min-1·m-2)reflux was significantly lower than that in children with grade Ⅱ(77.69±22.21 mL·min-1·m-2).The renal function compensation of contralateral non-reflux kidney increased in unilateral reflux group,which was higher than that in the control group and level Ⅱ,Ⅳ and Ⅴ of reflux group respectively.In VUR group of the same grade,sGFR decreased with the age at diagnosis.In unilateral grade V refux group,the tsGFR was lower than that in the unilateral grade I reflux group(133.51±48.21 vs.186.87+53.49mL·min-1·m-2).The patients with VUR of unilateral grade Ⅱ were significantly older than those with VUR of unilateral grades Ⅱ and Ⅳ.This study indicates that severe VUR is significantly associated with decreased renal function.Therefore,VUR should be diagnosed early and managed individually.
基金This project was supported by the National Natural Science Foundation of China(No.81873596).
文摘PKHD1 mutations are generally considered to cause autosomal recessive polycystic kidney disease(ARPKD).ARPKD is a rare disorder and one o f the most severe conditions leading to end-stage renal disease in childhood.With the biallelic deletion mutation,patients have difficulty in surviving the perinatal period,resulting in perinatal or neonatal death.This study retrospectively analyzed patient characteristics,imaging characteristics,laboratory examinations and family surveys from 7 Chinese children with different PKHD1 gene mutations diagnosed by high-throughput sequencing from January 2014 to February 2018.O f the 7 children,there were 3 males and 4 females.Eight missense mutations,two frameshift mutations,two deletion mutations,and two intronic slicing mutations were identified.Six of the mutations have not previously been identified.In the literature search,we identified a total of 29 Chinese children with PKHD1 mutations.The missense mutation c.2507T>C in exon 24 was found in one patient in our study,and five patients with liver fibrosis but normal renal function were reported in the literature.The missense mutation c.5935G>A in exon 37 was found in two patients in our study and three cases in the literature.Four patients had renal failure at an age as young as 1 year of those five patients with the missense mutation c.5935G>A in exon 37.It was concluded that:(1)Kidney length more than 2-3 SDs above the mean and early-onset hypertension might be associated with PKHDI-associated ARPICD;(2)The more enlarged the kidney size is,the lower the renal function is likely to be;(3)c.5935G>A may be a hot spot that leads to early renal failure in Chinese children with PKHD1 mutations;(4)c.2507T>C may be a hot-spot mutation associated with hepatic lesions in Chinese children with PKHD1.
基金supported by the National Natural Science Foundation of China(No.81873596).
文摘Objective:Autosomal dominant polycystic kidney disease(ADPKD)is mainly caused by the pathogenic mutation of PKD1 or PKD2 gene and usually affects bilateral kidneys.Synonymous mutations are generally assumed to be neutral as they do not alter amino acids.Herein,we described an extremely rare ADPKD child caused by a heterozygous synonymous mutation of PKD2 gene accompanied by massive proteinuria and congenital solitary kidney.Methods:Clinical characteristics of the patients were summarized.Whole-exome sequencing was performed to screen the disease-causing gene mutation,and reverse transcription polymerase chain reaction(RT-PCR)and Sanger sequencing were applied to analyze the impact of the identified mutation on gene transcription and splicing.Results:Polycystic changes were found in the solitary kidney of a girl initially presented with nephrotic-range proteinuria.Thereafter her mother and 2 other family members were diagnosed to be ADPKD.Whole-exome sequencing of the proband identified a heterozygous synonymous mutation(c.1716G>A,p.Lys572=)located in the splicing site of exon 7 in PKD2 gene,which was co-segregated with the PKD phenotype in the family.RT-PCR and direct sequencing of amplified products revealed that this heterozygous synonymous mutation led to exon7 skipping in PKD2 gene.Conclusion:We reported an extremely rare child case of ADPKD2 in combination with solitary kidney and nephrotic-range proteinuria,and firstly confirmed the pathogenicity of a heterozygous synonymous mutation(c.1716G>A)in PKD2 gene.The results indicate that synonymous mutations should not be excluded from disease-causing if they are located in splicing site of an exon.
文摘Objective Ubiquitin-specific protease 4(USP4)facilitates the development of transforming growth factor-beta 1(TGF-β1)-induced epithelial-mesenchymal transition(EMT)in various cancer cells.Moreover,EMT of renal tubular epithelial cells(RTECs)is required for the progression of renal interstitial fibrosis.However,the role of USP4 in EMT of RTECs remains unknown.The present study aimed to explore the effect of USP4 on the EMT of RTECs as well as the involved mechanism.Methods In established unilateral ureteral obstruction(UUO)rats and NRK-52E cells,immunohistochemistry and Western blot assays were performed.Results USP4 expression was increased significantly with obstruction time.In NRK-52E cells stimulated by TGF-β1,USP4 expression was increased in a time-dependent manner.In addition,USP4 silencing with specific siRNA indicated that USP4 protein was suppressed effectively.Meanwhile,USP4 siRNA treatment restored E-cadherin and weakened alpha smooth muscle actin(α-SMA)expression,indicating that USP4 may promote EMT.After treatment with USP4 siRNA and TGF-β1 for 24 h,the expression of TGF-β1 receptor type I(TβRI)was decreased.Conclusion USP4 promotes the EMT of RTECs through upregulating TβRI,thereby facilitating renal interstitial fibrosis.These findings may provide a potential target of USP4 in the treatment of renal fibrosis.
文摘Background: Current knowledge about clinical and genetic risk factors for aspirin-induced gastric mucosal injury is not sufficient to prevent these gastric mucosal lesions. Methods: We recruited aspirin takers as the exposed group and healthy volunteers as the control group. The exposed group was categorized into two subgroups such as subgroup A as gastric mucosal injury diagnosed by gastroscopy, including erosion, ulcer or bleeding of the esophagus, stomach, or duodenum; subgroup B as no injury of the gastric mucosa was detected by gastroscopy. Clinical information was collected, and 53 single nucleotide polymorphisms were evaluated. Results: Among 385 participants, 234 were in the aspirin-exposed group. According to gastroscopy, 82 belonged to subgroup A, 91 belonged to subgroup B, and gastroscopic results of 61 participants were not available. Using the Chi-square test and logistic regression, we found that peptic ulcer history (odds ratio [OR] = 5.924, 95% confidence intervals [C/]: 2.115-16.592), dual anti-platelet medication (OR = 3.443, 95% CI: 1.154-10.271 ), current Helicobacterpylori infection (OR = 2.242, 95% CI: 1.032-4.870), male gender (OR = 2.211, 95% CI: 1.027-4.760), GG genotype ofrs2243086 (OR = 4.516, 95% CI: I. 180-17.278), and AA genotype ofrs 1330344 (OR = 2.178, 95% CI: 1.016-4.669) were more frequent in subgroup A than subgroup B. In aspirin users who suffered from upper gastrointestinal bleeding, the frequency of the TT genotype ofrs2238631 and TT genotype ofrs2243100 was higher than in those without upper gastrointestinal bleeding. Conclusions: Peptic ulcer history, dual anti-platelet medication, tt. pylori current infection, and male gender were possible clinical risk factors for aspirin-induced gastric mucosal injury. GG genotype of rs2243086 and AA genotype of rs 1330344 were possible genetic risk factors. TT genotype ofrs2238631 and TT genotype ofrs2243100 may be risk factors for upper gastrointestinal bleeding in aspirin users.
基金supported by the National Natural Science Foundation of China(Grant No.30772360)the Natural Science Foundation of Health Department of Hubei Province,China(No.JX4B48).
文摘Hepatitis B virus X protein(HBx),a 17-kd protein encoded by X gene of hepatitis B virus(HBV),has been shown to function as a transcriptional trans-activator of a variety of viral and cellular promoter/enhancer elements.The aim of the study is to investigate the extracellular regulated protein kinases(ERKs)pathway of HBx on glomerular mesangial cell(GMC)proliferation and tumor necrosis factor-α(TNF-α)expression.The HBV X gene was amplified by polymerase chain reaction(PCR),inserted into the eukaryotic expression vector pCI-neo and confirmed by restriction endonuclease digestion and sequence analysis.PCI-neo containing HBV X gene(pCI-neo-X)was then transfected into cultured GMC line via liposome.GMC proliferation,TNF-αand its mRNA expression were compared in the condition of with or without U0126 in culture media.HBx,ERK1/2 and p-ERK1/2 expression in GMCs was assessed by Western blotting.TNF-αmRNA expression was assessed by semi-quantitative reverse transcription-PCR(RT-PCR).TNF-αlevel in supernatants was measured by ELISA.GMC proliferation was detected by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide(MTT)kit.The results showed that HBx expression was found in transfected GMCs and became prominent at 36th and 48th h after transfection whether with or without U0126 in culture media.TNF-αmRNA expression was significantly decreased in U0126 group compared with U0126-free group.TNF-αlevels in supernatants in PCI-neo-X transfection without U0126 group were(189.0�18.1)and(172.3�24.3)pg/mL at 36th and 48th h after transfec-tion,respectively.In contrast,TNF-αlevels in supernatants with U0126 were(65.6�11.6)and(84.0�24.6)pg/mL at 36th and 48th h,respectively.The TNF-αlevels in the latter groups were significantly lower than those in the former groups(P<0.05).GMCs proliferation was also lower in added U0126 group at 36th and 48th h after transfection.From above,we can conclude that HBx could induce GMC proliferation and increase TNF-αmRNA expression and its protein production.HBx upregulates TNF-αexpression and induces cell proliferation of GMC line partly through ERK1/2 signal transduction pathway.
基金This study was supported by National Natural Science Funding of China(81071891,81172209)Guangdong Provincial Science&Technology Funding(2010B0807017,2010B031600090).
文摘Objective:To explore the value of transrectal ultrasonography(TRUS)for tumor node metastasis(TNM)restaging for patients with locally advanced rectal cancer after neoadjuvant chemoradiotherapy(neo-CRT).Methods:One hundred and forty-nine patients with locally advanced rectal cancer(cT3-4 or cN+)who underwent TRUS after neo-CRT were retrospectively reviewed.TRUS restaging was compared with the results of post-operative pathological TNM findings.Results:After neo-CRT,the accuracy of TRUS for diagnosing T-staging was 30.9%,with 60.4%(90/149)of cases overestimated.The sensitivity of TRUS for T-staging(T0 vs T1 vs T2 vs T3 vs T4)were 16.3%,0%,12.5%,42.6%and 75.0%,respectively.The accuracy of TRUS for diagnosing N-staging after neo-CRT was 81.2%,with the sensitivities of N0 and N+were 93.3%and 31.0%,respectively.After neo-CRT,27.5%(41/149)of patients achieved pathologically complete response(pCR).The sensitivity,specificity,positive predictive value and negative predictive values of TRUS for pCR were 17.1%,99.1%,87.5%and 75.9%,respectively.Conclusions:TRUS can be applied for restaging T4 and N0,and has potential for screening out patients with pCR in those with locally advanced rectal cancer after neo-CRT,although some stages are overestimated for T-staging and its sensitivity for predicting pCR is low.