Research advances in the treatment of Crigler-Najjar syndrome / 临床肝胆病杂志

Crigler-Najjar syndrome (CNS) is an autosomal recessive disorder in which the content of plasma unconjugated bilirubin is increased due to the reduction or complete deficiency of the activity of bilirubin uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1), classified as CNS type Ⅰ and Ⅱ. CNS type Ⅰ is the most severe, which will develop into kernicterus, damage the brain nervous system, and even threaten the life of patients. This article introduces six CNS treatment techniques, including phototherapy, plasma exchange, drug therapy, liver transplantation, hepatocyte transplantation and gene therapy. The applicable patient types, treatment effects and existing deficiencies of each technique were summarized. Phototherapy, plasma exchange, drug therapy and hepatocyte transplantation can temporarily control serum levels and reduce the risk of jaundice, but cannot completely restore UGT1A1 enzyme activity; liver transplantation is currently the only treatment option for CNS type Ⅰ patients, but is limited by suitable liver donors and post-operative immune rejection. Gene therapy has the most promising application in the treatment of genetic disorders such as CNS, which can provide more viable therapeutic techniques for CNS patients.

Large-scale single-cell analysis reveals critical immune characteristics of COVID-19 patients

Dysfunctional immune response in the COVID-19 patients is a recurrent theme impacting symptoms and mortality, yet the detailed understanding of pertinent immune cells is not complete. We applied single-cell RNA sequencing to 284 samples from 205 COVID-19 patients and controls to create a comprehensive immune landscape. Lymphopenia and active T and B cell responses were found to coexist and associated with age, sex and their interactions with COVID-19. Diverse epithelial and immune cell types were observed to be virus-positive and showed dramatic transcriptomic changes. Elevation of ANXA1 and S100A9 in virus-positive squamous epithelial cells may enable the initiation of neutrophil and macrophage responses via the ANXA1-FPR1 and S100A8/9-TLR4 axes. Systemic upregulation of S100A8/A9, mainly by megakaryocytes and monocytes in the peripheral blood, may contribute to the cytokine storms frequently observed in severe patients. Our data provide a rich resource for understanding the pathogenesis and designing effective therapeutic strategies for COVID-19. HIGHLIGHTSO_LILarge-scale scRNA-seq analysis depicts the immune landscape of COVID-19 C_LIO_LILymphopenia and active T and B cell responses coexist and are shaped by age and sex C_LIO_LISARS-CoV-2 infects diverse epithelial and immune cells, inducing distinct responses C_LIO_LICytokine storms with systemic S100A8/A9 are associated with COVID-19 severity C_LI

SARS-CoV-2 Quasispecies provides insight into its genetic dynamics during infection

A novel coronavirus disease (COVID-19) caused by SARS-CoV-2 has been pandemic worldwide. The genetic dynamics of quasispecies afford RNA viruses a great fitness on cell tropism and host range. However, no quasispecies data of SARS-CoV-2 have been reported yet. To explore quasispecies haplotypes and its transmission characteristics, we carried out single-molecule real-time (SMRT) sequencing of the full-length of SARS-CoV-2 spike gene within 14 RNA samples from 2 infection clusters, covering first-to third-generation infected-patients. We observed a special quasispecies structure of SARS-CoV-2 (modeled as One-King): one dominant haplotype (mean abundance ~70.15%) followed by numerous minor haplotypes (mean abundance < 0.10%). We not only discovered a novel dominant haplotype of F1040 but also realized that minor quasispecies were also worthy of attention. Notably, some minor haplotypes (like F1040 and currently pandemic one G614) could potentially reveal adaptive and converse into the dominant one. However, minor haplotypes exhibited a high transmission bottleneck (~6% could be stably transmitted), and the new adaptive/dominant haplotypes were likely originated from genetic variations within a host rather than transmission. The evolutionary rate was estimated as 2.68-3.86 x 10-3 per site per year, which was larger than the estimation at consensus genome level. The One-King model and conversion event expanded our understanding of the genetic dynamics of SARS-CoV-2, and explained the incomprehensible phenomenon at the consensus genome level, such as limited cumulative mutations and low evolutionary rate. Moreover, our findings suggested the epidemic strains may be multi-host origin and future traceability would face huge difficulties.

Viral Kinetics and Antibody Responses in Patients with COVID-19

BackgroundA pandemic of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been spreading over the world. However, the viral dynamics, host serologic responses, and their associations with clinical manifestations, have not been well described in prospective cohort. MethodsWe conducted a prospective cohort and enrolled 67 COVID-19 patients admitting between Jan 26 and Feb 5, 2020. Clinical specimens including nasopharyngeal swab, sputum, blood, urine and stool were tested periodically according to standardized case report form with final follow-up on February 27. The routes and duration of viral shedding, antibody response, and their associations with disease severity and clinical manifestations were systematically evaluated. Coronaviral particles in clinical specimens were observed by transmission electron microscopy (TEM). ResultsThe median duration of SARS-CoV-2 RNA shedding were 12 (3-38), 19 (5-37), and 18 (7-26) days in nasopharyngeal swabs, sputum and stools, respectively. Only 13 urines (5.6%) and 12 plasmas (5.7%) were viral positive. Prolonged viral shedding was observed in severe patients than that of non-severe patients. Cough but not fever, aligned with viral shedding in clinical respiratory specimens, meanwhile the positive stool-RNA appeared to align with the proportion who concurrently had cough and sputum production, but not diarrhea. Typical coronaviral particles could be found directly in sputum by TEM. The anti-nucleocapsid-protein IgM started on day 7 and positive rate peaked on day 28, while that of IgG was on day 10 and day 49 after illness onset. IgM and IgG appear earlier, and their titers are significantly higher in severe patients than non-severe patients (p<0.05). The weak responders for IgG had a significantly higher viral clearance rate than that of strong responders (p= 0.011). ConclusionsNasopharyngeal, sputum and stools rather than blood and urine, were the major shedding routes for SARS-CoV-2, and meanwhile sputum had a prolonged viral shedding. Symptom cough seems to be aligned with viral shedding in clinical respiratory and fecal specimens. Stronger antibody response was associated with delayed viral clearance and disease severity. Summary boxesO_ST_ABSWhat is already known on this topicC_ST_ABSAs a newly appearing infectious disease, early efforts have focused on virus identification, describing the epidemiologic characteristics, clinical course, prognostics for critically illed cases and mortality. Among COVID-19 cases reported in mainland China (72 314 cases, updated through February 11, 2020), 81% are mild, 14% are severe, and 5% are critical. The estimated overall case fatality rate (CFR) is 2.3%. Some case series reported had shown that SARS-CoV-2 could shed in upper/lower respiratory specimens, stools, blood and urines of patients. However, important knowledge gaps remain, particularly regarding full kinetics of viral shedding and host serologic responses in association with clinical manifestations and host factors. What this study addsThe incubation period has no change after spreading out of Wuhan, and has no sex or age differences, however, children had prolonged incubation period. Due to early recognition and intervention, COVID-19 illness of Chongqing cohort is milder than that of Wuhan patients reported. This prospective cohort study on SARS-CoV-2 infection shows clearly that the viral and serological kinetics were related in duration of infection, disease severity, and clinical manifestations of COVID-19. Our data demonstrate that nasopharyngeal, sputum and stools are major shedding routes for SARS-CoV-2, and stronger NP antibody response is associated with delayed viral clearance and disease severity.

Diagnosis of Acute Respiratory Syndrome Coronavirus 2 Infection by Detection of Nucleocapsid Protein

BACKGROUNDNucleic acid test and antibody assay have been employed in the diagnosis for SARS-CoV-2 infection, but the use of viral antigen for diagnosis has not been successfully developed. Theoretically, viral antigen is the specific marker of the virus and precedes antibody appearance within the infected population. There is a clear need of detection of viral antigen for rapid and early diagnosis. METHODSWe included a cohort of 239 participants with suspected SARS-CoV-2 infection from 7 centers for the study. We measured nucleocapsid protein in nasopharyngeal swab samples in parallel with the nucleic acid test. Nucleic acid test was taken as the reference standard, and statistical evaluation was taken in blind. We detected nucleocapsid protein in 20 urine samples in another center, employing nasopharyngeal swab nucleic acid test as reference standard. RESULTSWe developed a fluorescence immunochromatographic assay for detecting nucleocapsid protein of SARS-CoV-2 in nasopharyngeal swab sample and urine within 10 minutes. 100% of nucleocapsid protein positive and negative participants accord with nucleic acid test for same samples. Further, earliest participant after 3 days of fever can be identified by the method. In an additional preliminary study, we detected nucleocapsid protein in urine in 73.6% of diagnosed COVID-19 patients. CONCLUSIONSThose findings indicate that nucleocapsid protein assay is an accurate, rapid, early and simple method for diagnosis of COVID-19. Appearance of nucleocapsid protein in urine coincides our finding of the SARS-CoV-2 invading kidney and might be of diagnostic value.

Culture-Positive Spontaneous Ascitic Infection in Patients with Acute Decompensated Cirrhosis: Multidrug-Resistant Pathogens and Antibiotic Strategies

Functional status of hepatitis B virus-specific T cells in patients with chronic hepatitis B virus infection: Exhausted or silenced? / 临床肝胆病杂志

Hepatitis B virus (HBV)-specific T cells in patients with chronic HBV infection are currently acknowledged as exhausted T cells. This article summarizes previous studies which support this hypothesis and then points out several inconsistencies between this hypothesis and clinical observation. Based on recent research findings, it is pointed out that in patients with chronic HBV infection, HBV-specific T cells might be dominated by silenced, early-differentiated progenitor T cells. Also, this article analyzes the possible causes of the silenced status of those T cells.

Definition and diagnosis of acute-on-chronic liver failure: new knowledge and implication / 中华肝脏病杂志

Acute-on-chronic liver failure (ACLF) is a common critical and severe syndrome in patients with chronic liver diseases in China and other countries in the Asia-Pacific region. In recent years, both the Eastern and Western experts have defined ACLF as a new type of liver disease manifesting as a high 28-day mortality rate (>30%) and extensive systematic inflammatory response. ACLF has become a hot topic in the field of liver diseases. This article reviews the research advances in the definition and etiological spectrum of ACLF and discusses the inspirations of such new knowledge for future research.

FGL2 Gene Polymorphism among Patients Infected with HBV / 中华医院感染学杂志

OBJECTIVE To characterize the relationship between promoter of FGL2 gene(site-1285T/A)polymorphism and clinical subgroups infected with HBV. METHODS The genotype was analyzed by using method of polymerase chain reaction(PCR),enzyme restriction after PCR amplification,and agarose gel electrophoresis.Statistics were used ?~2 and Logistic regression. RESULTS There were no statistic genotype or allele frequency differences between any two subgroups(except acute hepatitis B and liver cancer). CONCLUSIONS Promoter of fgl2 gene(site1285T/A) polymorphism has no relation with HBV infection.

Mathematical model of regulating competitive dysbacteriosis with antibiotics / 第三军医大学学报

Objective To explore the conditions for the restoration of competitive dysbacteriosis with antibiotics. Methods The mathematical model of the two competitive floras was analyzed by the Qualitative Theory of Ordinary Differential Equations. Results Three different types of dysbacteriosis and their restoring conditions were obtained. Conclusion Different restoring schemes should be applied for the regulation of different types of dysbacteriosis. Misuse of antibiotics can not result in satisfactory therapeutic effect.

Association between disease susceptivity of chronic HBV infection and SNP G-944C in CⅡTA gene promoter Ⅳ / 第三军医大学学报

0.05). Conclusion The polymorphism of G-944C in CⅡTA gene promoter Ⅳ was associated with the susceptivity of chronic HBV infection, but was not associated with severity of diseases. The individuals with chronic HBV infection of CC genotype are of less possibility to develop chronic liver disease than those of other genotypes.

Effect of lamivudine on survival rate of patients with chronic severe hepatitis B / 第三军医大学学报

Objective To investigate the survival rate of patients with chronic severe hepatitis B (CSH) after lamivudine treatment. Method A matched retrospective cohort study using data on patients derived from a prospectively collected chronic severe hepatitis B database was conducted. The database included patients who received or did not receive lamivudine treatment from October, 1999 to December, 2003. The match was conducted according with 3 variables: sex, age and duration of disease. The match ratio was 1∶1. Cases were patients who received lamivudine treatment (n=103).Controls were patients who did not receive lamivudine treatment (n=103). All the patients were followed up and their median survival time and survival rate were compared and evaluated. Results The median survival time in lamivudine treatment group and control group were 85 and 35 d respectively. The survival rate for 3 years in treatment group and control group were 42.7% and 23.4% respectively. There was significance difference in survival rate between the 2 groups (P

Genome-wide analysis of DNA methylated status of CpG islands with the modified AIMS technology / 解放军医学杂志

Objective To conduct the global assessment of methylated DNA status of CpG islands. Methods The modified AIMS (amplification of inter-methylated sites) technology was adopted, which mainly included 3 groups of isoschizomers (methylation-sensitive and methylation-insensitive restricted enzymes). The comparison analysis of methylome among 3 concentrations (6%, 8%, 10%) of polyacrylamide with 8M urea denaturing sequencing gel and the digestion effect among 3 groups of isoschizomers were carried out. The results of personal molecular imager FX system and autoradiography were also compared. Results Genome-wide detection of methylome of CpG islands might be the technical basis for the subsequent analysis. The resolving power and scope bands in 6% polyacrylamide denaturing sequencing gel were superior to those in gels with the other two concentrations. 3 groups of isoschizomers could meet the need of genome-wide detection of methylome of CpG islands. Personal molecular imager FX system and autoradiography could be used together, and they compensated each other. Conclusions The optimized AIMS technology is a kind of high-throughput method to analyze methylome, which is simple, specific, and easy to handle. The 3 groups of isoschizomers can cover most of CpG islands for genome-wide detection and get the ideal results. Only a small quantity of genome DNA is enough to meet the need of clinical detection.