Objective To analyze the association between mutation（s） in preS region of HBV and hepatitis B disease progress in Chinese patients with genotype C chronic HBV infection. Methods Ninety-three patients with chronic g...Objective To analyze the association between mutation（s） in preS region of HBV and hepatitis B disease progress in Chinese patients with genotype C chronic HBV infection. Methods Ninety-three patients with chronic genotype C HBV infection, including 24 asymptomatic carriers （ASC）, 26 patients with chronic hepatitis B （CHB）, 22 patients with liver cirrhosis （LC） and 21 HCC patients were investigated. Levels of HBV DNA, HBeAg, alanine aminotransferase （ALT）, asparate transaminase （AST） were measured. HBV preS region was analyzed by PCR direct sequencing. Results The prevalence of preS T3098C and T53C mutations ofgenotype C HBV was significantly higher in LC and HCC patients than ASC and CHB patients. The rate ofT3098C mutation in ASC, CHB, LC, and HCC patients were 0.00% （0/24）, 3.85% （1/26）, 9.09% （2/22）, and 30.77% （8/22）, respectively （P=0.0015）, while the rate of T53C mutation was I2.50% （3/24）, 3.85% （1/26）, 40.91% （9/22）, and 42.31% （11/26）, respectively （P=0.0012）. Conclusion The frequency of genotype C HBV preS T3098C and T53C mutations is associated with hepatitis B infection progression.展开更多
Hepatitis B virus(HBV)genotype B and C are two major genotypes that are prevalent in Asia and differ in natural history and disease progression.The impact of HBV genotypes on viral replication and protein expression h...Hepatitis B virus(HBV)genotype B and C are two major genotypes that are prevalent in Asia and differ in natural history and disease progression.The impact of HBV genotypes on viral replication and protein expression has been explored by the transfection of hepatoma cells with replication-competent HBV DNA,which mimics the later stages of the viral life cycle.However,the influence of HBV genotypes on the early events of viral infection remains undetermined,mainly due to the difficulties in obtaining sufficient infectious viral particles for infection assays.Here,we report that a high-titer HBV inoculum can be generated from the transient transfection-based cell model after optimizing transfection conditions and modifying the HBV-expressing construct.By performing in vitro infection assays using transiently transfected derived viruses,we found that clinical genotype C isolates possessed higher infectivity than genotype B isolates.Moreover,we identified a naturally occurring mutation sL21S in small hepatitis B surface protein,which markedly decreased the infectivity of HBV genotype C isolates,but not that of genotype B isolates.In summary,using infectious viral particles provided by the optimized transient transfection-based cell model,we have been able to investigate a wide range of HBV variants on viral infectivity,which may contribute to our understanding of the reasons for different clinical outcomes in HBV infections and the development of therapeutic drugs targeting the early stages of HBV life cycle.展开更多
BACKGROUND The presence of two distinct hepatitis B virus(HBV)Pol RT polymorphisms,rt269L and rt269I,could contribute to the unique clinical or virological phenotype of HBV genotype C2.Therefore,a simple and sensitive...BACKGROUND The presence of two distinct hepatitis B virus(HBV)Pol RT polymorphisms,rt269L and rt269I,could contribute to the unique clinical or virological phenotype of HBV genotype C2.Therefore,a simple and sensitive method capable of identifying both types in chronic hepatitis B(CHB)patients infected with genotype C2 should be developed.AIM To develop a novel simple and sensitive locked nucleic acid(LNA)-real timepolymerase chain reaction(RT-PCR)method capable of identifying two rt269 types in CHB genotype C2 patients.METHODS We designed proper primer and probe sets for LNA-RT-PCR for the separation of rt269 types.Using synthesized DNAs of the wild type and variant forms,melting temperature analysis,detection sensitivity,and endpoint genotyping for LNA-RT-PCR were performed.The developed LNA-RT-PCR method was applied to a total of 94 CHB patients of genotype C2 for the identification of two rt269 polymorphisms,and these results were compared with those obtained by a direct sequencing protocol.RESULTS The LNA-RT-PCR method could identify two rt269L and rt269I polymorphisms of three genotypes,two rt269L types[‘L1’(WT)and‘L2’]and one rt269I type(‘I’)in single(63 samples,72.4%)or mixed forms(24 samples,27.6%)in 87(92.6%sensitivity)of 94 samples from Korean CHB patients.When the results were compared with those obtained by the direct sequencing protocol,the LNA-RT-PCR method showed the same results in all but one of 87 positive detected samples(98.9%specificity).CONCLUSION The newly developed LNA-RT-PCR method could identify two rt269 polymorphisms,rt269L and rt269I,in CHB patients with genotype C2 infections.This method could be effectively used for the understanding of disease progression in genotype C2 endemic areas.展开更多
Bovine parainfluenza virus type 3（BPIV3） is considered as one of the most important respiratory tract pathogens of both young and adult cattle, and widespread among cattle in the world. BPIV3 was first reported in C...Bovine parainfluenza virus type 3（BPIV3） is considered as one of the most important respiratory tract pathogens of both young and adult cattle, and widespread among cattle in the world. BPIV3 was first reported in China in 2008 and four strains of BPIV3 were isolated from Shandong Province, known as genotype C（BPIV3c）. Pathogen investigations had shown that BPIV3 c infection was very common among cattle in China. To date, BPIV3 can be classified into genotypes A, B and C based on genetic and phylogenetic analysis. Serological survey also demonstrates that BPIV3 infection is widespread in China, however, there is still no available vaccine for BPIV3 prevention in China nowadays. In the present study, the BPIV3 c strain SD0835 was continuously passaged on Madin-Darby bovine kidney（MDBK） cells for hundreds of times, and the pathogenicity of passage 209 was reduced in guinea pigs. The passage 209 of BPIV3 c strain SD0835 was used as a live vaccine candidate to immunize the guinea pigs. The vaccination results revealed that two vaccinations could induce excellent serum neutralizing antibody responses as well as proliferation of T lymphocytes. The vaccinated guinea pigs were well protected against challenge with a low passage of BPIV3 c strain SD0835. Additionally, the percentages of CD4~＋ and CD8~＋ T cell subsets of animals in vaccinated group increased after immunization; T cell subsets on day 2 after challenge in both groups decreased, and the decline of CD4~＋ and CD8~＋ T cell subsets levels of four guinea pigs in vaccinated group was relatively moderate, comparing with that of the control group. These data support further testing of the attenuated virus as an effective candidate vaccine.展开更多
With pegylated interferon and ribavirin, more than half of all chronically-infected hepatitis C patients can achieve a sustained virologic response; however, patients with genotype 1 infections and those with other po...With pegylated interferon and ribavirin, more than half of all chronically-infected hepatitis C patients can achieve a sustained virologic response; however, patients with genotype 1 infections and those with other poor prognostic factors have relatively inferior treatment response rates. Since new therapies are still years away from approval, it is incumbent upon providers to maximize the therapeutic efficacy of today＇s treatment. The later the virus is undetectable in serum during treatment, the less likely it will be eradicated. Patients with a delayed or slow virologic response to therapy （at least a 2-1og10 decrease in baseline hepatitis C RNA yet detectable viremia at 12 wk of therapy and undetectable virus 12 wk subsequently） may, therefore, benefit from an extended therapy course beyond one of standard duration. Although higher rates of treatment discontinuation may plague this approach, 72 wk of treatment for genotype 1-infected slow-responders may improve response rates and diminish relapse rates relative to those of 48 wk. Based on data from both viral kinetic and clinical studies, therapy prolongation in slow responders may be a reasonable strategy to improve response rates in these treatment-refractory patients.展开更多
AIM： TO estimate the prevalence of the lactase non-persistent genotype （C/C-23910） in a northern Russian population in accordance with ethnicity, and to evaluate self-reported milk consumption depending on lactase ...AIM： TO estimate the prevalence of the lactase non-persistent genotype （C/C-23910） in a northern Russian population in accordance with ethnicity, and to evaluate self-reported milk consumption depending on lactase activity. METHODS： Blood samples for genotyping lactase activity, defining the C/T-13910 variant by polymerase chain reaction, and direct sequencing were taken from 231 medical students of Russian origin aged 17-26 years. We analyzed milk product consumption by questionnaire which was specially designed for the estimation of milk consumption and abdominal complaints. RESULTS： We found that the prevalence of the C/C-13190 genotype in the northern Russian population was 35.6%. The other genotypes nearby C/T-13910 and associated with lactase activity were not present in the study population. The consumption of milk among people with the non-persistent genotype tended to be lower than among the lactose tolerant subjects, but was not statistically significant. CONCLUSION： An investigation of the lactase persistent genotype in a northern Russian population has not been performed before, The genotype did not affect the consumption of milk products in this population which could be explained by low consumption of milk products among the entire study population.展开更多
BACKGROUND Hepatitis C virus genotype 3a(HCV G3a)is highly prevalent in Pakistan.Due to the elevated cost of available Food and Drug Administration-approved drugs against HCV,medicinal natural products of potent antiv...BACKGROUND Hepatitis C virus genotype 3a(HCV G3a)is highly prevalent in Pakistan.Due to the elevated cost of available Food and Drug Administration-approved drugs against HCV,medicinal natural products of potent antiviral activity should be screened for the cost-effective treatment of the disease.Furthermore,from natural products,active compounds against vital HCV proteins like non-structural protein 3(NS3)protease could be identified to prevent viral proliferation in the host.AIM To develop cost-effective HCV genotype 3a NS3 protease inhibitors from citrus fruit extracts.METHODS Full-length NS3 without co-factor non-structural protein 4A(NS4A)and codon optimized NS3 protease in fusion with NS4A were expressed in Escherichia coli.The expressed protein was purified by metal ion affinity chromatography and gel filtration.Citrus fruit extracts were screened using fluorescence resonance energy transfer(FRET)assay against the protease and polyphenols were identified as potential inhibitors using electrospray ionization-mass spectrometry(MS)/MS technique.Among different polyphenols,highly potent compounds were screened using molecular modeling approaches and consequently the most active compound was further evaluated against HCV NS4A-NS3 protease domain using FRET assay.RESULTS NS4A fused with NS3 protease domain gene was overexpressed and the purified protein yield was high in comparison to the lower yield of the full-length NS3 protein.Furthermore,in enzyme kinetic studies,NS4A fused with NS3 protease proved to be functionally active compared to full-length NS3.So it was concluded that co-factor NS4A fusion is essential for the purification of functionally active protease.FRET assay was developed and validated by the half maximal inhibitory concentration(IC50)values of commercially available inhibitors.Screening of citrus fruit extracts against the native purified fused NS4A-NS3 protease domain showed that the grapefruit mesocarp extract exhibits the highest percentage inhibition 91%of protease activity.Among the compounds identified by LCMS analysis,hesperidin showed strong binding affinity with the protease catalytic triad having S-score value of-10.98.CONCLUSION Fused NS4A-NS3 protease is functionally more active,which is effectively inhibited by hesperidin from the grapefruit mesocarp extract with an IC50 value of 23.32μmol/L.展开更多
Host genetic factors may predict the outcome and treatment response in hepatitis C virus（HCV）infection.One of these factors is the single nucleotide polymorphisms of the interleukin 28B（IL28B）gene.We sought to eva...Host genetic factors may predict the outcome and treatment response in hepatitis C virus（HCV）infection.One of these factors is the single nucleotide polymorphisms of the interleukin 28B（IL28B）gene.We sought to evaluate the outcome of pegylated interferon and ribavirin therapy in association with IL-28B rs8099917 and rsl2980275 in patients infected with HCV genotype 4.A total of 180 patients with chronic hepatitis C were selected from Egyptians who have received combined therapy with pegylated interferon and ribavirin for 6 months and their response was evaluated after follow-up at 0,6,12,24 and 48 weeks from the beginning of the therapy.Blood samples were collected from responders and non-responders.Genomic DNA was extracted from whole blood and genotyping was carried out by polymerase chain reaction and restriction fragment length polymorphism（PCR-RFLP）.Our results showed that TT genotype of rs8099917 was associated with higher sustained viral response（SVR）rates and G allele represented a risk factor for failure of response（OR=3.7,CI=1.8：7.64）while rs12980275 was not significantly associated with SVR in genotype 4 Egyptian patients.The determination of 1L-28B SNPs may be useful in enhancing correct prediction of SVR achievement in treating this group of genotype 4 patients.展开更多
AIM To evaluate magnitude/direction of changes in peripheral lipid profiles in patients undergoing direct acting therapy for hepatitis C by genotype.METHODS Mono-infected patients with hepatitis C were treated with gu...AIM To evaluate magnitude/direction of changes in peripheral lipid profiles in patients undergoing direct acting therapy for hepatitis C by genotype.METHODS Mono-infected patients with hepatitis C were treated with guideline-based DAAs at a university-based liver clinic. Patient characteristics and laboratory values were collected before and after the treatment period. Baseline demographics included age, ethnicity, hypertension, diabetes, hyperlipidemia, treatment regimen, and fibrosis stage. Total cholesterol(TCHOL), high density lipoprotein(HDL), low density lipoprotein(LDL), triglycerides(TG), and liver function tests were measured prior to treatment and ETR. Changes in lipid and liver function were evaluated by subgroups with respect to genotype. Mean differences were calculated for each lipid profile and liver function component(direction/magnitude). The mean differences in lipid profiles were then compared between genotypes for differences in direction/magnitude. Lipid profile and liver function changes were evaluated with Levene's test and student's t test. Mean differences in lipid profiles were compared between genotypes using ANOVA, post hoc analysis via the Bonferroni correction or Dunnett T3.RESULTS Three hundred and seventy five patients enrolled with 321(85.6%) achieving sustained-viral response at 12 wk. 72.3% were genotype 1(GT1), 18.1% genotype 2(GT2), 9.7% genotype 3(GT3). Baseline demographics were similar. Significant change in lipid profiles were seen with GT1 and GT3(ΔGT1, p and ΔGT3, p), with TCHOL increasing(+5.3, P = 0.005 and +16.1, P < 0.001), HDL increasing(+12.5, P < 0.001 and +7.9, P = 0.038), LDL increasing(+7.4, P = 0.058 and +12.5, P < 0.001), and TG decreasing(-5.9, P = 0.044 and-9.80 P = 0.067). Among genotypes(ΔGT1 v. ΔGT2 v. ΔGT3, ANOVA), significant mean differences were seen with TCHOL(+5.3 v. +0.1 v. +16.1, P = 0.017) and HDL(+12.3 v. +2 v. +7.9, P = 0.040). Post-hoc, GT3 was associated with a greater increase in TCHOL than GT1 and GT2(P = 0.028 and P = 0.019).CONCLUSION Successful DAA therapy results in increases in TCHOL, LDL, and HDL and decrease in TG, particularly in GT1/GT3. Changes are most pronounced in GT3.展开更多
Enterovirus 71 （EV71） is a common cause of Hand, foot, and mouth disease （HFMD） and may also cause severe neurological diseases, such as encephalitis and poliomyelitis-like paralysis. To examine the genetic divers...Enterovirus 71 （EV71） is a common cause of Hand, foot, and mouth disease （HFMD） and may also cause severe neurological diseases, such as encephalitis and poliomyelitis-like paralysis. To examine the genetic diversity of EV71, we determined and analyzed the complete VP1 sequences （891 nueleotides） from nine EV71 strains isolated in Fuyang, China. We found that nine EV71 strains isolated were over 98% homologous at the nucleotide level and 93%-100% homologous tO members of the C4 subgenogroup. At the amino acid level, these Fuyang strains were 99% -100% homologous to one another, 97%-100% homologous to members of the C4 subgenogroup, and the histidine（H） at amino acid position 22 was conserved among the Fuyang strains. The results indicate that Fuyang isolates belong to genotype C4, and an H at position 22 appears to be a marker for the Fuyang strains.展开更多
Hepatitis C virus(HCV)infection represents a major public health issue.Hepatitis C can be cured bytherapy,but many infected individuals are unaware of their status.Effective HCV screening,fast diagnosis and characteri...Hepatitis C virus(HCV)infection represents a major public health issue.Hepatitis C can be cured bytherapy,but many infected individuals are unaware of their status.Effective HCV screening,fast diagnosis and characterization,and hepatic fibrosis staging are highly relevant for controlling transmission,treating infected patients and,consequently,avoiding end-stage liver disease.Exposure to HCV can be determined with high sensitivity and specificity with currently available third generation serology assays.Additionally,the use of point-of-care tests can increase HCV screening opportunities.However,active HCV infection must be confirmed by direct diagnosis methods.Additionally,HCV genotyping is required prior to starting any treatment.Increasingly,high-volume clinical laboratories use different types of automated platforms,which have simplified sample processing,reduced hands-on-time,minimized contamination risks and human error and ensured full traceability of results.Significant advances have also been made in the field of fibrosis stage assessment with the development of non-invasive methods,such as imaging techniques and serum-based tests.However,no single test is currently available that is able to completely replace liver biopsy.This review focuses on approved commercial tools used to diagnose HCV infection and the recommended hepatic fibrosis staging tests.展开更多
AIMTo evaluate and compare the efficacy and safety of telaprevir(TVR)-and simeprevir(SMV)-based triple therapies in elderly patients,specifically patients aged 66 years or older.METHODSThe present study enrolled 112 a...AIMTo evaluate and compare the efficacy and safety of telaprevir(TVR)-and simeprevir(SMV)-based triple therapies in elderly patients,specifically patients aged 66 years or older.METHODSThe present study enrolled 112 and 76 Japanese patients with chronic hepatitis C virus genotype 1b infection who were treated with a 12-wk TVR-based or SMV-based triple therapy,respectively,followed by a dual therapy that included pegylated interferonαand ribavirin(RBV)for 12 wk.The patients were categorized into two groups according to age as follows:A younger group of patients aged≤65 years old and an older group of patients aged>65 years old.Among the patients treated with TVR-based triple therapy,34 patients were included in the older group.The median ages were 56 years(range:28-65 years)in the younger group and 69 years(range:66-81 years)in the older group.Among the patients treated with SMV-based triple therapy,39 patients were included in the older group.The median ages were 59 years(range:36-65 years)in the younger group and 71 years(range:66-86 years)in the older group.The clinical,biochemical and virological data were analyzed before and during treatment.RESULTSAmong the patients treated with the TVR-based triple therapy,no significant difference in the sustained virological response(SVR)was found between the younger(80.8%)and older(88.2%)groups.The SVR rates for patients with the interleukin 28B(IL28B)(rs8099917)TG/GG-genotypes(73.9%and 60.0%in the younger and older groups,respectively)were significantly lower than for patients with the IL28B TT-genotype(86.3%and 92.9%,respectively).The cumulative exposure to RBV for the entire 24-wk treatment period(as a percentage of the target dose)was significantly higher in the younger group than in the older group(91.7%vs 66.7%,respectively,P vs 81.9%,respectively).A multivariate analysis identified the TT-genotype of IL28B(OR=8.160;95%CI:1.593-41.804,P=0.012)and the adherence of RBV(>60%)(OR=11.052;95%CI:1.160-105.273,P=0.037)as independent factors associated with the SVR.Adverse events resulted in discontinuation of the treatment in 11.3%and 14.7%of the younger and older groups,respectively.Among the patients treated with the SMV-based triple therapy,no significant difference in the SVR rare was found between the younger(81.1%)and older(82.1%)groups.The SVR rates for patients with the IL28B TG/GG-genotypes(77.8%and 64.7%in the younger and older groups,respectively)were significantly lower than for patients with the IL28B TT-genotype(88.2%and 100%,respectively).A multivariate analysis identified the TT-genotype of IL28B as an independent factor associated with the SVR(OR=9.677;95%CI:1.114-84.087,P=0.040).Adverse events resulted in discontinuation of the treatment in 7.0%and 14.3%of patients in the younger and older groups,respectively.CONCLUSIONBoth TVR-and SMV-based triple therapies can be successfully used to treat patients aged 66 years or older with genotype 1b chronic hepatitis C.Genotyping of the IL28B indicates a potential to achieve SVR in these difficult-to-treat elderly patients.展开更多
BACKGROUND Chronic pulmonary aspergillosis(CPA)is a rare syndrome that is often accompanied by gradual lung tissue destruction.Voriconazole is usually employed as the first-line agent for CPA treatment.However,some pa...BACKGROUND Chronic pulmonary aspergillosis(CPA)is a rare syndrome that is often accompanied by gradual lung tissue destruction.Voriconazole is usually employed as the first-line agent for CPA treatment.However,some patients can develop hepatotoxicity and often were forced to stop voriconazole treatment.AIM To record the improving trend of liver function and the therapeutic effects in patients after lowering the trough concentration of voriconazole.METHODS This study retrospectively analyzed 12 adult CPA patients who developed hepatotoxicity during the voriconazole treatment.In these patients,the oral dose was reduced to 3/4 or 1/2 of the standard dose(4 mg/kg,twice daily),and the lower limit of voriconazole trough concentration was maintained more than 0.5μg/m L.The trend of remission of liver toxicity after drug reduction in 12 patients was recorded.During the same period,25 patients who received standard doses served as the control group.Data from the two groups were collected and analyzed for different parameters such as demographic characteristics,underlying pulmonary disorders,laboratory tests,and therapeutic effect.The differences between the two groups were statistically compared.RESULTS Hepatotoxicity occurred in 12 patients within 28-65 d after oral voriconazole treatment.Hepatotoxicity was mainly manifested by the significantly increased level of gamma-glutamyltransferase and a slight increase of alanine aminotransferase and aspartate aminotransferase.The oral dose of voriconazole was reduced to approximately 3 mg/kg in seven patients and approximately 2 mg/kg in five patients.The average trough concentrations for the 12 patients before and after voriconazole oral dose reduction were 3.17±1.47μg/m L(1.5-6.0μg/m L)and 1.70±0.78μg/m L(0.6-3.3μg/m L),respectively(P=0.02).After lowering the trough concentrations,the hepatotoxicity was alleviated in all the patients.However,gamma-glutamyltransferase levels declined slowly.After 4 mo of treatment,7 of the 12 patients were successfully treated in the low trough concentrations group(41.7%).Similarly,8 of the 25 patients in the standard treatment dose group(32.0%)were effectively treated.There was no statistical difference between the groups(P=0.72).CONCLUSION Reducing the lower limit of the voriconazole trough concentration to 0.5μg/m L can alleviate the hepatotoxicity and maintained certain clinical efficacy in CPA patients;however,patients should be closely monitored.展开更多
AIM: To study milk consumption and subjective milk- related symptoms in adults genotyped for adult-type hypolactasia. METHODS: A total of 1900 Finnish adults were genotyped for the C/T-13910 variant of adult-type hypo...AIM: To study milk consumption and subjective milk- related symptoms in adults genotyped for adult-type hypolactasia. METHODS: A total of 1900 Finnish adults were genotyped for the C/T-13910 variant of adult-type hypolactasia and filled in a structured questionnaire concerning milk consumption and gastrointestinal problems. RESULTS: The C/C-13910 genotype of adult-type hypolactasia was present in 18% of the study population. The prevalence of the C/C-13910 genotype was higher among subjects who were undergoing investigations because of abdominal symptoms (24%, P < 0.05). Those with the C/C-13910 genotype drank less milk than subjects with either the C/T-13910 or the T/T-13910 genotype of lactase persistence (18% vs 38%; 18% vs 36%, P < 0.01). Subjects with the C/C-13910 genotype had experienced more gastrointestinal symptoms (84%) during the preceding three-month period than those with the C/T-13910 (79%, P < 0.05) or the T/T-13910 genotype (78 %, P < 0.05). Only 9% (29/338) of the subjects with the C/C-13910 genotype consumed milk and reported no symptoms from it.CONCLUSION: Gastrointestinal symptoms are more common among adults with the C/C-13910 genotype of adult-type hypolactasia than in those with genotypes of lactase persistence.展开更多
The CRISPR/Cas9 system has been tailored to a revolutionary genetic tool because of its remarkable simplicity and efficacy.While complex genome editing in the mouse since the 1990 s has been dominated by the use of em...The CRISPR/Cas9 system has been tailored to a revolutionary genetic tool because of its remarkable simplicity and efficacy.While complex genome editing in the mouse since the 1990 s has been dominated by the use of embryonic stem（ES） cells,CRISPR/Cas9 now offers a versatile and fast approach to precisely modify virtually any DNA regions directly in mouse zygotes.Yet,this relative simplicity does not preclude a conscientious preparatory work that is often neglected when initiating a project.Here,we describe the key steps leading to successful generation of a double knockout（KO） mouse by simultaneously targeting two homolog genes,Tmem176 a and Tmem176 b,which are located in the same genomic locus.Additionally,we show that similar efficiency can be obtained in a mixed genetic background or directly in the C57BL/6 inbred strain.Thus,presented as a detailed case study that should be helpful to the non-specialists,we focus on the genotyping strategy to anticipate the various possibilities.展开更多
AIM： To correlate the C/T-13910 variant, associated with lactase persistence/non-persistence （adulttype hypolactasia） trait, with intestinal disaccharidase activities in different age groups of the adult population...AIM： To correlate the C/T-13910 variant, associated with lactase persistence/non-persistence （adulttype hypolactasia） trait, with intestinal disaccharidase activities in different age groups of the adult population.METHODS： Intestinal biopsies were obtained from 222 adults aged 18 to 83 years undergoing upper gastrointestinal endoscopy because of unspecified abdominal complaints. The biopsies were assayed for lactase, sucrase and maltase activities and genotyped for the C/T-13910 variant using PCR-minisequencing. RESULTS： There was a significant correlation between lactase activity and the C/T-13910 variant （P 〈 0.00001）. The mean level of lactase activity among subjects with C/C-1391o genotype was 6.86± 0.35 U/g, with C/T-13910 genotype 37.8 ± 1.4 U/g, and with T/T-13910 genotype 57.6± 2.4 U/g protein, showing a trimodal distribution of this enzyme activity. Significant differences were also observed in maltase activities among individuals with different C/T-13910 genotypes （P = 0.005）. In contrast, in sucrase activity, no significant differences emerged between the C/T-13910 genotypes （P = 0.14）. There were no statistical differences in lactase （P = 0.84）, sucrase （P = 0.18）, or maltase activity （P = 0.24） among different age groups. In the majority （〉 84%） of the patients with the C/C-13910 genotype associated with lactase non- persistence, the lactase activity was less than 10 U/g protein.CONCLUSION： Our study demonstrates a statistically significant correlation between the C/T-13910 genotype and lactase activity and this correlation is not affected by age in adults but the cut-off value of 20 U/g protein used for the diagnosis of lactase non-persistence might be too high.展开更多
Hepatitis B virus(HBV)infection is a global health problem and more than 350 million people worldwide are chronic carriers of the virus.Despite the recent dramatic decline in HBV chronic patients through successful pr...Hepatitis B virus(HBV)infection is a global health problem and more than 350 million people worldwide are chronic carriers of the virus.Despite the recent dramatic decline in HBV chronic patients through successful programs of hepatitis B surface antigen vaccination,South Korea is still recognized as an endemic area of HBV infection.HBV infections in South Korea exhibit several distinct features in epidemiologic and clinical aspects.In this review paper,we summarize the distinct HBV mutation patterns related to clinical severity and the molecular epidemiologic traits in Korean chronic patients based on previous reports.Generally,several lines of evidence,including our previous results,have led to the conclusion that a combination of the exclusive predominance of genotype C2,which is prone to mutations,the high prevalence of basal core promoter double mutations,and the presence of distinct immune responses against HBV proteins in the Korean population may generate the distinct HBV variants rarely or not encountered in other areas,which results in distinct clinical manifestations in Korean chronic patients.This may provide a novel insight into the relationships between clinical severity,HBV genotype distribution,and HBV naturally occurring variants.展开更多
Our previous studies demonstrated that oral vitamin A supplementation during late-stage pregnancy and the neonatal stage enhances birth weight,growth performance,and mRNA expression related to muscle and preadipocyte ...Our previous studies demonstrated that oral vitamin A supplementation during late-stage pregnancy and the neonatal stage enhances birth weight,growth performance,and mRNA expression related to muscle and preadipocyte development in beef cattle.The alcohol dehydrogenase 1C(ADH1C)c.-64T>C genotype also correlated with vitamin A concentration in beef production.This study aimed to investigate the effects of vitamin A supplementation on the muscle development and vitamin A metabolism in weaned beef calves with different ADH1C genotypes.Twenty male calves(90 d of age;initial BW:89.03 kg[SD 8.60])were stratified according to ADH1C genotype and vitamin A treatment(duration:3 months)and randomly assigned to 4 groups with a 22 factorial arrangement.Vitamin A treatments included the following:control(10,000 IU/kg of as-fed,a.TT type;b.TC type);treatment(40,000 IU/kg of as-fed,c.TT type;and d.TC type).Parameters including BW,FI,blood,longissimus dorsi muscle,and liver status during the experimental period were analyzed using the generalized linear model(GLM)procedure and Tukey's test by SAS 9.4 program.Serum vitamin A was significantly increased(P<0.05)in the vitamin A treatment group at 4 and 6 months of age.TT type calves showed higher serum vitamin A concentration(P<0.05)than the TC type calves.Serum triglyceride and non-esterified fatty acid(NEFA)levels increased(P<0.05)in the treatment group compared with the control at 6 months of age.However,BW,ADG and FI showed no differences between the groups.In addition,mRNA expression in longissimus dorsi muscle revealed upregulation of paired box 7(PAX7)(P<0.05)after the vitamin A treatment period based on biopsy results.Both ADH1C and aldehyde dehydrogenase(ALDH)1A1 mRNA expression was downregulated(P<0.01)by vitamin A supplementation.The TC type of ADH1C showed higher mRNA expression than the TT type.However,no effect was observed on adipogenic mRNA expression(preadipocyte factor-1[PREF-1],peroxisome proliferator-activated receptor gamma[PPARg],fatty acid binding protein 4[FABP4])in all groups.Our findings suggest that weaned calves treated with vitamin A may promote the storage of satellite cells by elevating PAX7 gene expression in the muscle.The TC type calves may show increased capacity for vitamin A metabolism,which can be used in genetically customizing feed management to maximize beef production in the calves.展开更多
基金supported by the financial grants from Beijing Municipal Science & Technology Commission (D08050702870000)Basic Research Project ("973"Project:2005CB523104)+3 种基金the National Projects on the Control of Major Infectious Diseases (Grant no.2008ZX10002-012)supported by the financial grants from Beijing Municipal Science & Technology Commission (D0805700650805)the National Projects on Major Infectious Diseases, Ministry of Science and Technology of China (No.2008ZX10002-12No.2008ZX10002-004)
文摘Objective To analyze the association between mutation（s） in preS region of HBV and hepatitis B disease progress in Chinese patients with genotype C chronic HBV infection. Methods Ninety-three patients with chronic genotype C HBV infection, including 24 asymptomatic carriers （ASC）, 26 patients with chronic hepatitis B （CHB）, 22 patients with liver cirrhosis （LC） and 21 HCC patients were investigated. Levels of HBV DNA, HBeAg, alanine aminotransferase （ALT）, asparate transaminase （AST） were measured. HBV preS region was analyzed by PCR direct sequencing. Results The prevalence of preS T3098C and T53C mutations ofgenotype C HBV was significantly higher in LC and HCC patients than ASC and CHB patients. The rate ofT3098C mutation in ASC, CHB, LC, and HCC patients were 0.00% （0/24）, 3.85% （1/26）, 9.09% （2/22）, and 30.77% （8/22）, respectively （P=0.0015）, while the rate of T53C mutation was I2.50% （3/24）, 3.85% （1/26）, 40.91% （9/22）, and 42.31% （11/26）, respectively （P=0.0012）. Conclusion The frequency of genotype C HBV preS T3098C and T53C mutations is associated with hepatitis B infection progression.
基金supported by grants of the National Youth Natural Science Foundation of China,China(82102380)the Youth Natural Science Foundation of Jiangsu Province(BK20200126)the National Youth Natural Science Foundation of China,China(81602400)。
文摘Hepatitis B virus(HBV)genotype B and C are two major genotypes that are prevalent in Asia and differ in natural history and disease progression.The impact of HBV genotypes on viral replication and protein expression has been explored by the transfection of hepatoma cells with replication-competent HBV DNA,which mimics the later stages of the viral life cycle.However,the influence of HBV genotypes on the early events of viral infection remains undetermined,mainly due to the difficulties in obtaining sufficient infectious viral particles for infection assays.Here,we report that a high-titer HBV inoculum can be generated from the transient transfection-based cell model after optimizing transfection conditions and modifying the HBV-expressing construct.By performing in vitro infection assays using transiently transfected derived viruses,we found that clinical genotype C isolates possessed higher infectivity than genotype B isolates.Moreover,we identified a naturally occurring mutation sL21S in small hepatitis B surface protein,which markedly decreased the infectivity of HBV genotype C isolates,but not that of genotype B isolates.In summary,using infectious viral particles provided by the optimized transient transfection-based cell model,we have been able to investigate a wide range of HBV variants on viral infectivity,which may contribute to our understanding of the reasons for different clinical outcomes in HBV infections and the development of therapeutic drugs targeting the early stages of HBV life cycle.
基金Supported by the National Research Foundation of Korea,No.2022R1A2B5B01001421the Korea Health Technology R&D Project through the Korea Health Industry Development Institute,the Ministry of Health&Welfare,Republic of Korea,No.HI22C0476.
文摘BACKGROUND The presence of two distinct hepatitis B virus(HBV)Pol RT polymorphisms,rt269L and rt269I,could contribute to the unique clinical or virological phenotype of HBV genotype C2.Therefore,a simple and sensitive method capable of identifying both types in chronic hepatitis B(CHB)patients infected with genotype C2 should be developed.AIM To develop a novel simple and sensitive locked nucleic acid(LNA)-real timepolymerase chain reaction(RT-PCR)method capable of identifying two rt269 types in CHB genotype C2 patients.METHODS We designed proper primer and probe sets for LNA-RT-PCR for the separation of rt269 types.Using synthesized DNAs of the wild type and variant forms,melting temperature analysis,detection sensitivity,and endpoint genotyping for LNA-RT-PCR were performed.The developed LNA-RT-PCR method was applied to a total of 94 CHB patients of genotype C2 for the identification of two rt269 polymorphisms,and these results were compared with those obtained by a direct sequencing protocol.RESULTS The LNA-RT-PCR method could identify two rt269L and rt269I polymorphisms of three genotypes,two rt269L types[‘L1’(WT)and‘L2’]and one rt269I type(‘I’)in single(63 samples,72.4%)or mixed forms(24 samples,27.6%)in 87(92.6%sensitivity)of 94 samples from Korean CHB patients.When the results were compared with those obtained by the direct sequencing protocol,the LNA-RT-PCR method showed the same results in all but one of 87 positive detected samples(98.9%specificity).CONCLUSION The newly developed LNA-RT-PCR method could identify two rt269 polymorphisms,rt269L and rt269I,in CHB patients with genotype C2 infections.This method could be effectively used for the understanding of disease progression in genotype C2 endemic areas.
基金funded by a grant from the National Natural Science Foundation of China(31372452)a fund for Science and Technology Plan from Harbin Science and Technology Bureau,Heilongjiang Province,China(2012AA6BN020)a grant from the National Key Technologies R&D Program of China during the 12th Five-Year Plan period(2012BAD12B03-3)
文摘Bovine parainfluenza virus type 3（BPIV3） is considered as one of the most important respiratory tract pathogens of both young and adult cattle, and widespread among cattle in the world. BPIV3 was first reported in China in 2008 and four strains of BPIV3 were isolated from Shandong Province, known as genotype C（BPIV3c）. Pathogen investigations had shown that BPIV3 c infection was very common among cattle in China. To date, BPIV3 can be classified into genotypes A, B and C based on genetic and phylogenetic analysis. Serological survey also demonstrates that BPIV3 infection is widespread in China, however, there is still no available vaccine for BPIV3 prevention in China nowadays. In the present study, the BPIV3 c strain SD0835 was continuously passaged on Madin-Darby bovine kidney（MDBK） cells for hundreds of times, and the pathogenicity of passage 209 was reduced in guinea pigs. The passage 209 of BPIV3 c strain SD0835 was used as a live vaccine candidate to immunize the guinea pigs. The vaccination results revealed that two vaccinations could induce excellent serum neutralizing antibody responses as well as proliferation of T lymphocytes. The vaccinated guinea pigs were well protected against challenge with a low passage of BPIV3 c strain SD0835. Additionally, the percentages of CD4~＋ and CD8~＋ T cell subsets of animals in vaccinated group increased after immunization; T cell subsets on day 2 after challenge in both groups decreased, and the decline of CD4~＋ and CD8~＋ T cell subsets levels of four guinea pigs in vaccinated group was relatively moderate, comparing with that of the control group. These data support further testing of the attenuated virus as an effective candidate vaccine.
文摘With pegylated interferon and ribavirin, more than half of all chronically-infected hepatitis C patients can achieve a sustained virologic response; however, patients with genotype 1 infections and those with other poor prognostic factors have relatively inferior treatment response rates. Since new therapies are still years away from approval, it is incumbent upon providers to maximize the therapeutic efficacy of today＇s treatment. The later the virus is undetectable in serum during treatment, the less likely it will be eradicated. Patients with a delayed or slow virologic response to therapy （at least a 2-1og10 decrease in baseline hepatitis C RNA yet detectable viremia at 12 wk of therapy and undetectable virus 12 wk subsequently） may, therefore, benefit from an extended therapy course beyond one of standard duration. Although higher rates of treatment discontinuation may plague this approach, 72 wk of treatment for genotype 1-infected slow-responders may improve response rates and diminish relapse rates relative to those of 48 wk. Based on data from both viral kinetic and clinical studies, therapy prolongation in slow responders may be a reasonable strategy to improve response rates in these treatment-refractory patients.
基金The Sigrid Jusélius Foundation,Helsinki,Finland and Tampere University Hospital Research Funds
文摘AIM： TO estimate the prevalence of the lactase non-persistent genotype （C/C-23910） in a northern Russian population in accordance with ethnicity, and to evaluate self-reported milk consumption depending on lactase activity. METHODS： Blood samples for genotyping lactase activity, defining the C/T-13910 variant by polymerase chain reaction, and direct sequencing were taken from 231 medical students of Russian origin aged 17-26 years. We analyzed milk product consumption by questionnaire which was specially designed for the estimation of milk consumption and abdominal complaints. RESULTS： We found that the prevalence of the C/C-13190 genotype in the northern Russian population was 35.6%. The other genotypes nearby C/T-13910 and associated with lactase activity were not present in the study population. The consumption of milk among people with the non-persistent genotype tended to be lower than among the lactose tolerant subjects, but was not statistically significant. CONCLUSION： An investigation of the lactase persistent genotype in a northern Russian population has not been performed before, The genotype did not affect the consumption of milk products in this population which could be explained by low consumption of milk products among the entire study population.
文摘BACKGROUND Hepatitis C virus genotype 3a(HCV G3a)is highly prevalent in Pakistan.Due to the elevated cost of available Food and Drug Administration-approved drugs against HCV,medicinal natural products of potent antiviral activity should be screened for the cost-effective treatment of the disease.Furthermore,from natural products,active compounds against vital HCV proteins like non-structural protein 3(NS3)protease could be identified to prevent viral proliferation in the host.AIM To develop cost-effective HCV genotype 3a NS3 protease inhibitors from citrus fruit extracts.METHODS Full-length NS3 without co-factor non-structural protein 4A(NS4A)and codon optimized NS3 protease in fusion with NS4A were expressed in Escherichia coli.The expressed protein was purified by metal ion affinity chromatography and gel filtration.Citrus fruit extracts were screened using fluorescence resonance energy transfer(FRET)assay against the protease and polyphenols were identified as potential inhibitors using electrospray ionization-mass spectrometry(MS)/MS technique.Among different polyphenols,highly potent compounds were screened using molecular modeling approaches and consequently the most active compound was further evaluated against HCV NS4A-NS3 protease domain using FRET assay.RESULTS NS4A fused with NS3 protease domain gene was overexpressed and the purified protein yield was high in comparison to the lower yield of the full-length NS3 protein.Furthermore,in enzyme kinetic studies,NS4A fused with NS3 protease proved to be functionally active compared to full-length NS3.So it was concluded that co-factor NS4A fusion is essential for the purification of functionally active protease.FRET assay was developed and validated by the half maximal inhibitory concentration(IC50)values of commercially available inhibitors.Screening of citrus fruit extracts against the native purified fused NS4A-NS3 protease domain showed that the grapefruit mesocarp extract exhibits the highest percentage inhibition 91%of protease activity.Among the compounds identified by LCMS analysis,hesperidin showed strong binding affinity with the protease catalytic triad having S-score value of-10.98.CONCLUSION Fused NS4A-NS3 protease is functionally more active,which is effectively inhibited by hesperidin from the grapefruit mesocarp extract with an IC50 value of 23.32μmol/L.
文摘Host genetic factors may predict the outcome and treatment response in hepatitis C virus（HCV）infection.One of these factors is the single nucleotide polymorphisms of the interleukin 28B（IL28B）gene.We sought to evaluate the outcome of pegylated interferon and ribavirin therapy in association with IL-28B rs8099917 and rsl2980275 in patients infected with HCV genotype 4.A total of 180 patients with chronic hepatitis C were selected from Egyptians who have received combined therapy with pegylated interferon and ribavirin for 6 months and their response was evaluated after follow-up at 0,6,12,24 and 48 weeks from the beginning of the therapy.Blood samples were collected from responders and non-responders.Genomic DNA was extracted from whole blood and genotyping was carried out by polymerase chain reaction and restriction fragment length polymorphism（PCR-RFLP）.Our results showed that TT genotype of rs8099917 was associated with higher sustained viral response（SVR）rates and G allele represented a risk factor for failure of response（OR=3.7,CI=1.8：7.64）while rs12980275 was not significantly associated with SVR in genotype 4 Egyptian patients.The determination of 1L-28B SNPs may be useful in enhancing correct prediction of SVR achievement in treating this group of genotype 4 patients.
文摘AIM To evaluate magnitude/direction of changes in peripheral lipid profiles in patients undergoing direct acting therapy for hepatitis C by genotype.METHODS Mono-infected patients with hepatitis C were treated with guideline-based DAAs at a university-based liver clinic. Patient characteristics and laboratory values were collected before and after the treatment period. Baseline demographics included age, ethnicity, hypertension, diabetes, hyperlipidemia, treatment regimen, and fibrosis stage. Total cholesterol(TCHOL), high density lipoprotein(HDL), low density lipoprotein(LDL), triglycerides(TG), and liver function tests were measured prior to treatment and ETR. Changes in lipid and liver function were evaluated by subgroups with respect to genotype. Mean differences were calculated for each lipid profile and liver function component(direction/magnitude). The mean differences in lipid profiles were then compared between genotypes for differences in direction/magnitude. Lipid profile and liver function changes were evaluated with Levene's test and student's t test. Mean differences in lipid profiles were compared between genotypes using ANOVA, post hoc analysis via the Bonferroni correction or Dunnett T3.RESULTS Three hundred and seventy five patients enrolled with 321(85.6%) achieving sustained-viral response at 12 wk. 72.3% were genotype 1(GT1), 18.1% genotype 2(GT2), 9.7% genotype 3(GT3). Baseline demographics were similar. Significant change in lipid profiles were seen with GT1 and GT3(ΔGT1, p and ΔGT3, p), with TCHOL increasing(+5.3, P = 0.005 and +16.1, P < 0.001), HDL increasing(+12.5, P < 0.001 and +7.9, P = 0.038), LDL increasing(+7.4, P = 0.058 and +12.5, P < 0.001), and TG decreasing(-5.9, P = 0.044 and-9.80 P = 0.067). Among genotypes(ΔGT1 v. ΔGT2 v. ΔGT3, ANOVA), significant mean differences were seen with TCHOL(+5.3 v. +0.1 v. +16.1, P = 0.017) and HDL(+12.3 v. +2 v. +7.9, P = 0.040). Post-hoc, GT3 was associated with a greater increase in TCHOL than GT1 and GT2(P = 0.028 and P = 0.019).CONCLUSION Successful DAA therapy results in increases in TCHOL, LDL, and HDL and decrease in TG, particularly in GT1/GT3. Changes are most pronounced in GT3.
基金Scientific Research Fund of Institute of Pathogen Biology(2008IPB108)
文摘Enterovirus 71 （EV71） is a common cause of Hand, foot, and mouth disease （HFMD） and may also cause severe neurological diseases, such as encephalitis and poliomyelitis-like paralysis. To examine the genetic diversity of EV71, we determined and analyzed the complete VP1 sequences （891 nueleotides） from nine EV71 strains isolated in Fuyang, China. We found that nine EV71 strains isolated were over 98% homologous at the nucleotide level and 93%-100% homologous tO members of the C4 subgenogroup. At the amino acid level, these Fuyang strains were 99% -100% homologous to one another, 97%-100% homologous to members of the C4 subgenogroup, and the histidine（H） at amino acid position 22 was conserved among the Fuyang strains. The results indicate that Fuyang isolates belong to genotype C4, and an H at position 22 appears to be a marker for the Fuyang strains.
基金Supported by A Miguel Servet contract No.MS09/00044 funded by FIS-ISCIII(Spanish Government)to MartróEgrant PI10/01734 within the"Plan Nacional de I+D+I"co-financed by"ISCIII-Subdirección General de Evaluación y el Fondo Eu-ropeo de Desarrollo Regional"(FEDER)to González V,Saludes V,MartróE
文摘Hepatitis C virus(HCV)infection represents a major public health issue.Hepatitis C can be cured bytherapy,but many infected individuals are unaware of their status.Effective HCV screening,fast diagnosis and characterization,and hepatic fibrosis staging are highly relevant for controlling transmission,treating infected patients and,consequently,avoiding end-stage liver disease.Exposure to HCV can be determined with high sensitivity and specificity with currently available third generation serology assays.Additionally,the use of point-of-care tests can increase HCV screening opportunities.However,active HCV infection must be confirmed by direct diagnosis methods.Additionally,HCV genotyping is required prior to starting any treatment.Increasingly,high-volume clinical laboratories use different types of automated platforms,which have simplified sample processing,reduced hands-on-time,minimized contamination risks and human error and ensured full traceability of results.Significant advances have also been made in the field of fibrosis stage assessment with the development of non-invasive methods,such as imaging techniques and serum-based tests.However,no single test is currently available that is able to completely replace liver biopsy.This review focuses on approved commercial tools used to diagnose HCV infection and the recommended hepatic fibrosis staging tests.
基金Supported by Grants-in-Aid for Scientific Research(C)(to Yamagiwa S)from Japan Society for the Promotion of Science(JSPS),No.15K08991
文摘AIMTo evaluate and compare the efficacy and safety of telaprevir(TVR)-and simeprevir(SMV)-based triple therapies in elderly patients,specifically patients aged 66 years or older.METHODSThe present study enrolled 112 and 76 Japanese patients with chronic hepatitis C virus genotype 1b infection who were treated with a 12-wk TVR-based or SMV-based triple therapy,respectively,followed by a dual therapy that included pegylated interferonαand ribavirin(RBV)for 12 wk.The patients were categorized into two groups according to age as follows:A younger group of patients aged≤65 years old and an older group of patients aged>65 years old.Among the patients treated with TVR-based triple therapy,34 patients were included in the older group.The median ages were 56 years(range:28-65 years)in the younger group and 69 years(range:66-81 years)in the older group.Among the patients treated with SMV-based triple therapy,39 patients were included in the older group.The median ages were 59 years(range:36-65 years)in the younger group and 71 years(range:66-86 years)in the older group.The clinical,biochemical and virological data were analyzed before and during treatment.RESULTSAmong the patients treated with the TVR-based triple therapy,no significant difference in the sustained virological response(SVR)was found between the younger(80.8%)and older(88.2%)groups.The SVR rates for patients with the interleukin 28B(IL28B)(rs8099917)TG/GG-genotypes(73.9%and 60.0%in the younger and older groups,respectively)were significantly lower than for patients with the IL28B TT-genotype(86.3%and 92.9%,respectively).The cumulative exposure to RBV for the entire 24-wk treatment period(as a percentage of the target dose)was significantly higher in the younger group than in the older group(91.7%vs 66.7%,respectively,P vs 81.9%,respectively).A multivariate analysis identified the TT-genotype of IL28B(OR=8.160;95%CI:1.593-41.804,P=0.012)and the adherence of RBV(>60%)(OR=11.052;95%CI:1.160-105.273,P=0.037)as independent factors associated with the SVR.Adverse events resulted in discontinuation of the treatment in 11.3%and 14.7%of the younger and older groups,respectively.Among the patients treated with the SMV-based triple therapy,no significant difference in the SVR rare was found between the younger(81.1%)and older(82.1%)groups.The SVR rates for patients with the IL28B TG/GG-genotypes(77.8%and 64.7%in the younger and older groups,respectively)were significantly lower than for patients with the IL28B TT-genotype(88.2%and 100%,respectively).A multivariate analysis identified the TT-genotype of IL28B as an independent factor associated with the SVR(OR=9.677;95%CI:1.114-84.087,P=0.040).Adverse events resulted in discontinuation of the treatment in 7.0%and 14.3%of patients in the younger and older groups,respectively.CONCLUSIONBoth TVR-and SMV-based triple therapies can be successfully used to treat patients aged 66 years or older with genotype 1b chronic hepatitis C.Genotyping of the IL28B indicates a potential to achieve SVR in these difficult-to-treat elderly patients.
文摘BACKGROUND Chronic pulmonary aspergillosis(CPA)is a rare syndrome that is often accompanied by gradual lung tissue destruction.Voriconazole is usually employed as the first-line agent for CPA treatment.However,some patients can develop hepatotoxicity and often were forced to stop voriconazole treatment.AIM To record the improving trend of liver function and the therapeutic effects in patients after lowering the trough concentration of voriconazole.METHODS This study retrospectively analyzed 12 adult CPA patients who developed hepatotoxicity during the voriconazole treatment.In these patients,the oral dose was reduced to 3/4 or 1/2 of the standard dose(4 mg/kg,twice daily),and the lower limit of voriconazole trough concentration was maintained more than 0.5μg/m L.The trend of remission of liver toxicity after drug reduction in 12 patients was recorded.During the same period,25 patients who received standard doses served as the control group.Data from the two groups were collected and analyzed for different parameters such as demographic characteristics,underlying pulmonary disorders,laboratory tests,and therapeutic effect.The differences between the two groups were statistically compared.RESULTS Hepatotoxicity occurred in 12 patients within 28-65 d after oral voriconazole treatment.Hepatotoxicity was mainly manifested by the significantly increased level of gamma-glutamyltransferase and a slight increase of alanine aminotransferase and aspartate aminotransferase.The oral dose of voriconazole was reduced to approximately 3 mg/kg in seven patients and approximately 2 mg/kg in five patients.The average trough concentrations for the 12 patients before and after voriconazole oral dose reduction were 3.17±1.47μg/m L(1.5-6.0μg/m L)and 1.70±0.78μg/m L(0.6-3.3μg/m L),respectively(P=0.02).After lowering the trough concentrations,the hepatotoxicity was alleviated in all the patients.However,gamma-glutamyltransferase levels declined slowly.After 4 mo of treatment,7 of the 12 patients were successfully treated in the low trough concentrations group(41.7%).Similarly,8 of the 25 patients in the standard treatment dose group(32.0%)were effectively treated.There was no statistical difference between the groups(P=0.72).CONCLUSION Reducing the lower limit of the voriconazole trough concentration to 0.5μg/m L can alleviate the hepatotoxicity and maintained certain clinical efficacy in CPA patients;however,patients should be closely monitored.
基金the Sigrid Jusélius Foundation, Helsinki, Finlandthe Foundation for Nutrition Research, Helsinki, Finland+3 种基金the Research Foundation of Alfred Kordelin, Helsinki, FinlandHelsinki University Hospital Research Funding, Helsinki, Finlandthe Foundation for Promoting Occupational Medicine in Finland,Helsinki,Finlandthe Academy of Finland
文摘AIM: To study milk consumption and subjective milk- related symptoms in adults genotyped for adult-type hypolactasia. METHODS: A total of 1900 Finnish adults were genotyped for the C/T-13910 variant of adult-type hypolactasia and filled in a structured questionnaire concerning milk consumption and gastrointestinal problems. RESULTS: The C/C-13910 genotype of adult-type hypolactasia was present in 18% of the study population. The prevalence of the C/C-13910 genotype was higher among subjects who were undergoing investigations because of abdominal symptoms (24%, P < 0.05). Those with the C/C-13910 genotype drank less milk than subjects with either the C/T-13910 or the T/T-13910 genotype of lactase persistence (18% vs 38%; 18% vs 36%, P < 0.01). Subjects with the C/C-13910 genotype had experienced more gastrointestinal symptoms (84%) during the preceding three-month period than those with the C/T-13910 (79%, P < 0.05) or the T/T-13910 genotype (78 %, P < 0.05). Only 9% (29/338) of the subjects with the C/C-13910 genotype consumed milk and reported no symptoms from it.CONCLUSION: Gastrointestinal symptoms are more common among adults with the C/C-13910 genotype of adult-type hypolactasia than in those with genotypes of lactase persistence.
基金supported by the Labex IGO project(n°ANR11-LABX-0016-01)funded by the "Investissements d'Avenir" French Government program,managed by the French National Research Agency(ANR)+1 种基金the context of the IHU-Cesti project(EXT173947) which received French government financial support managed by the National Research Agency via the "Investment Into The Future program" ANR-10-IBHU-005supported by Nantes Metropole and Region Pays de la Loire. C.L.was supported by Fondation Progreffe
文摘The CRISPR/Cas9 system has been tailored to a revolutionary genetic tool because of its remarkable simplicity and efficacy.While complex genome editing in the mouse since the 1990 s has been dominated by the use of embryonic stem（ES） cells,CRISPR/Cas9 now offers a versatile and fast approach to precisely modify virtually any DNA regions directly in mouse zygotes.Yet,this relative simplicity does not preclude a conscientious preparatory work that is often neglected when initiating a project.Here,we describe the key steps leading to successful generation of a double knockout（KO） mouse by simultaneously targeting two homolog genes,Tmem176 a and Tmem176 b,which are located in the same genomic locus.Additionally,we show that similar efficiency can be obtained in a mixed genetic background or directly in the C57BL/6 inbred strain.Thus,presented as a detailed case study that should be helpful to the non-specialists,we focus on the genotyping strategy to anticipate the various possibilities.
基金a grant from the Emil Aaltonen Foundation, Tampere, the Sigrid Jusélius Foundation, Helsinki, the Center of Excellence in Disease Genetics of the Academy of Finland, Helsinki University Research Funding, Helsinki, and Finnish Cultural Foundation, Helsinki, Finland
文摘AIM： To correlate the C/T-13910 variant, associated with lactase persistence/non-persistence （adulttype hypolactasia） trait, with intestinal disaccharidase activities in different age groups of the adult population.METHODS： Intestinal biopsies were obtained from 222 adults aged 18 to 83 years undergoing upper gastrointestinal endoscopy because of unspecified abdominal complaints. The biopsies were assayed for lactase, sucrase and maltase activities and genotyped for the C/T-13910 variant using PCR-minisequencing. RESULTS： There was a significant correlation between lactase activity and the C/T-13910 variant （P 〈 0.00001）. The mean level of lactase activity among subjects with C/C-1391o genotype was 6.86± 0.35 U/g, with C/T-13910 genotype 37.8 ± 1.4 U/g, and with T/T-13910 genotype 57.6± 2.4 U/g protein, showing a trimodal distribution of this enzyme activity. Significant differences were also observed in maltase activities among individuals with different C/T-13910 genotypes （P = 0.005）. In contrast, in sucrase activity, no significant differences emerged between the C/T-13910 genotypes （P = 0.14）. There were no statistical differences in lactase （P = 0.84）, sucrase （P = 0.18）, or maltase activity （P = 0.24） among different age groups. In the majority （〉 84%） of the patients with the C/C-13910 genotype associated with lactase non- persistence, the lactase activity was less than 10 U/g protein.CONCLUSION： Our study demonstrates a statistically significant correlation between the C/T-13910 genotype and lactase activity and this correlation is not affected by age in adults but the cut-off value of 20 U/g protein used for the diagnosis of lactase non-persistence might be too high.
基金Supported by A National Research Foundation of Korea grant funded by the Korean government(MEST)No.2013005810
文摘Hepatitis B virus(HBV)infection is a global health problem and more than 350 million people worldwide are chronic carriers of the virus.Despite the recent dramatic decline in HBV chronic patients through successful programs of hepatitis B surface antigen vaccination,South Korea is still recognized as an endemic area of HBV infection.HBV infections in South Korea exhibit several distinct features in epidemiologic and clinical aspects.In this review paper,we summarize the distinct HBV mutation patterns related to clinical severity and the molecular epidemiologic traits in Korean chronic patients based on previous reports.Generally,several lines of evidence,including our previous results,have led to the conclusion that a combination of the exclusive predominance of genotype C2,which is prone to mutations,the high prevalence of basal core promoter double mutations,and the presence of distinct immune responses against HBV proteins in the Korean population may generate the distinct HBV variants rarely or not encountered in other areas,which results in distinct clinical manifestations in Korean chronic patients.This may provide a novel insight into the relationships between clinical severity,HBV genotype distribution,and HBV naturally occurring variants.
基金the National Research Foundation of Korea(NRF)grant funded by the Korea government(MSIT)(2020R1A2B5B02001843).
文摘Our previous studies demonstrated that oral vitamin A supplementation during late-stage pregnancy and the neonatal stage enhances birth weight,growth performance,and mRNA expression related to muscle and preadipocyte development in beef cattle.The alcohol dehydrogenase 1C(ADH1C)c.-64T>C genotype also correlated with vitamin A concentration in beef production.This study aimed to investigate the effects of vitamin A supplementation on the muscle development and vitamin A metabolism in weaned beef calves with different ADH1C genotypes.Twenty male calves(90 d of age;initial BW:89.03 kg[SD 8.60])were stratified according to ADH1C genotype and vitamin A treatment(duration:3 months)and randomly assigned to 4 groups with a 22 factorial arrangement.Vitamin A treatments included the following:control(10,000 IU/kg of as-fed,a.TT type;b.TC type);treatment(40,000 IU/kg of as-fed,c.TT type;and d.TC type).Parameters including BW,FI,blood,longissimus dorsi muscle,and liver status during the experimental period were analyzed using the generalized linear model(GLM)procedure and Tukey's test by SAS 9.4 program.Serum vitamin A was significantly increased(P<0.05)in the vitamin A treatment group at 4 and 6 months of age.TT type calves showed higher serum vitamin A concentration(P<0.05)than the TC type calves.Serum triglyceride and non-esterified fatty acid(NEFA)levels increased(P<0.05)in the treatment group compared with the control at 6 months of age.However,BW,ADG and FI showed no differences between the groups.In addition,mRNA expression in longissimus dorsi muscle revealed upregulation of paired box 7(PAX7)(P<0.05)after the vitamin A treatment period based on biopsy results.Both ADH1C and aldehyde dehydrogenase(ALDH)1A1 mRNA expression was downregulated(P<0.01)by vitamin A supplementation.The TC type of ADH1C showed higher mRNA expression than the TT type.However,no effect was observed on adipogenic mRNA expression(preadipocyte factor-1[PREF-1],peroxisome proliferator-activated receptor gamma[PPARg],fatty acid binding protein 4[FABP4])in all groups.Our findings suggest that weaned calves treated with vitamin A may promote the storage of satellite cells by elevating PAX7 gene expression in the muscle.The TC type calves may show increased capacity for vitamin A metabolism,which can be used in genetically customizing feed management to maximize beef production in the calves.
