Objective To learn the significance of detection of high sensitive troponin T and creatine kinase isoenzyme in diagnosis of pediatric myocarditis.To provide reliable laboratory diagnosis method for the disease.Methods 23 cases of pediatric myocarditis,28 cases of viral myocarditis with capillary bronchitis and 61.cases of myocarditis with neonatal pneumonia were selected as the research objects;and 48 cases of healthy control group,55 cases of capillary bronchitis and 49 cases of neonatal pneumonia were also selected.Blood samples were collected from all the patients and healthy controls,and the levels of high sensitive serum troponin T and creatine kinase isoenzyme were also measured.Results There was no significant difference in the detection results of high sensitive troponin T and creatine kinase isoenzyme between the healthy control group,children with capillary bronchitis and neonatal pneumonia (all P ＞ 0.05);high sensitive troponin T and creatine kinase isoenzyme detection results of myocarditis,myocarditis complicated with bronchiolitis,myocarditis complicated with neonatal pneumonia were higher than those in healthy control group,the differences were statistically significant (t =13.723,6.628,10.079,9.475,17.650,15.364,all P ＜ 0.05).The abnormal rates of combined detection of children with myocarditis,myocarditis combined with capillary bronchitis,myocarditis combined with neonatal pneumonia were higher than those of single detection of high sensitive troponin T and single detection of creatine kinase isoenzyme (x2 =7.426,6.310,6.720,4.308,4.381,6.900,all P ＜0.05).The high sensitive troponin T and creatine kinase isoenzyme in the children with the age of 1-12 months and 1-3 were lower than those with the age of ＜ 1 month,the differences were statistically significant (t =3.498,4.043,4.202,4.132,all P ＜ 0.05).Conclusion The simultaneous detection of high sensitive troponin T and creatine kinase isoenzyme can be used in the diagnosis of pediatric myocarditis,with good clinical application value.

Objective To study the biosynthesis of the second metabolites in Aquilaria sinensis.Methods Chromones of agarwood were elicited by tissue culture and the inducer contents were determined by HPLC.Results 2-(2-Phenylethyl) chromones could be elicited in excised lateral roots suspension culture of A.sinensis.Conclusion Fungal extracts of Menanotus flavolives could induce the production of characteristic components of agarwood in A.sinensis.

Objective To investigate the related clinical factors and homology of strains in Stenotrophomonas maltophilia (S. Maltophilia) infections in 15 patients with liver transplantation. Methods Fifteen patients with S. Maltophilia infection from September to December 2006 were enrolled and their clinical data were collected. Minimal inhibitory concentrations (MICs) of 10 antimierobial agents against S. Maltophilia were determined by Etest strips. Antibiogram was carried out by resistance analysis assembly with WHONET 5 software. The genomic DNA of all the isolates was digested with Xbal and subjected to pulse-field gel electrophoresis (PFGE). Results All patients received mechanical ventilation during the treatment and had a history of long-term use of extended-spectrum β-lactamases and quinolones. MICs of 10 antimicrobial agents indicated that S. Maltophilia were susceptible to several antimicrobial agents including compound sulfamethoxazole and ciprofloxacin, but the best active agent against these resistant isolates was minocycline in vitro. The results of all 15 S. Maltophilia antibiograms were accordance with PFGE patterns. All 15 S. Maltophilia isolates were classified as 2 PFGE patterns: 9 for pattern A and 6 for pattern B. Conclusion Mechanical ventilation might be associated with the S. Maltophilia septicemia in patients with liver transplantation.

Objective To determine the changes of peripheral levels of T helper cell cytokines of patients with chronic hepatitis B (CHB) during antiviral treatment,and to further explore its clinical significance.Methods The plasma levels of interleukin (IL)-2,IL-6,IL-10,interferon (IFN)-γ and tumor necrosis factor(TNF)-α of thirty-three CHB patients during antiviral treatment (entecavir) were measured using enzyme linked immunosorbont assay (ELISA).And their biochemical indicators of liver function were determined.The differences of cytokines levels before and after antiviral treatment were compared using ANOVA.The correlations between the changes of cytokines and alanine transaminase (ALT),hepatitis B virus (HBV) DNA levels were analyzed.Results Levels of IFNγ before and 12,24,48 weeks after treatment were (5.98±2.77),(5.95±3.37),(2.93±2.15) and (9.29±4.65) pg/mL,respectively (F=3.845,P＜0.05),which were positively correlated with ALT levels (r =0.396,P＜0.05).Both TNF-α and IL-10 levels declined after antiviral treatment,which were significantly different at different time points (F=20.156 and 16.695,respectively; both P＜0.05),and both levels of TNF-α and IL-10 were positively correlated with ALT levels (r=0.354and 0.316,respectively; both P＜0.05) and positively correlated with HBV DNA levels (r=0.382and 0.386,respectively; both P＜0.05).While both IL-2 and IL-6 levels were not significantly different between before and after antiviral treatment (F=0.010 and 0.932,respectively; both P＞0.05).The serum levels of ALT and HBV DNA before and after antiviral treatment were all significantly different (F=17.69 and 198.98,respectively; both P＜0.05),which declined gradually during treatment and were positively correlated (r =0.581,P＜0.05).Conclusions IL-10,IFNγand TNF-α may be involved in the pathologic process of CHB,and closely related to the deterioration of the disease.Monitoring plasma levels of these cytokines during antiviral treatment could be useful to understand the immune status and evaluate the efficacy of antiviral drugs.

Various liver diseases result in terminal hepatic failure, and liver transplantation, cell transplantation and artificial liver support systems are emerging as effective therapies for severe hepatic disease. However, all of these treatments are limited by organ or cell resources, so developing a sufficient number of functional hepatocytes for liver regeneration is a priority. Liver regeneration is a complex process regulated by growth factors (GFs), cytokines, transcription factors (TFs), hormones, oxidative stress products, metabolic networks, and microRNA. It is well-known that the function of isolated primary hepatocytes is hard to maintain; when cultured in vitro, these cells readily undergo dedifferentiation, causing them to lose hepatocyte function. For this reason, most studies focus on inducing stem cells, such as embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), hepatic progenitor cells (HPCs), and mesenchymal stem cells (MSCs), to differentiate into hepatocyte-like cells (HLCs) in vitro. In this review, we mainly focus on the nature of the liver regeneration process and discuss how to maintain and enhance in vitro hepatic function of isolated primary hepatocytes or stem cell-derived HLCs for liver regeneration. In this way, hepatocytes or HLCs may be applied for clinical use for the treatment of terminal liver diseases and may prolong the survival time of patients in the near future.
Cell Differentiation ; Embryonic Stem Cells ; cytology ; Hepatocytes ; cytology ; Humans ; Induced Pluripotent Stem Cells ; cytology ; Liver Regeneration ; Mesenchymal Stem Cells ; cytology ; Primary Cell Culture ; methods ; Regenerative Medicine ; methods ; Stem Cells ; cytology

Background::Hypertension and non-alcoholic fatty liver disease (NAFLD) share several pathophysiologic risk factors, and the exact relationship between the two remains unclear. Our study aims to provide evidence concerning the relationship between hypertension and NAFLD by analyzing data from the National Health and Nutrition Examination Survey (NHANES) 2017-2018 and Mendelian randomization (MR) analyses.Methods::Weighted multivariable-adjusted logistic regression was applied to assess the relationship between hypertension and NAFLD risk by using data from the NHANES 2017-2018. Subsequently, a two-sample MR study was performed using the genome-wide association study (GWAS) summary statistics to identify the causal association between hypertension, systolic blood pressure (SBP), diastolic blood pressure (DBP), and NAFLD. The primary inverse variance weighted (IVW) and other supplementary MR approaches were conducted to verify the causal association between hypertension and NAFLD. Sensitivity analyses were adopted to confirm the robustness of the results.Results::A total of 3144 participants were enrolled for our observational study in NHANES. Weighted multivariable-adjusted logistic regression analysis suggested that hypertension was positively related to NAFLD risk (odds ratio [OR] = 1.677; 95% confidence interval [CI], 1.159-2.423). SBP ≥130 mmHg and DBP ≥80 mmHg were also significantly positively correlated with NAFLD. Moreover, hypertension was independently connected with liver steatosis ( β = 7.836 [95% CI, 2.334-13.338]). The results of MR analysis also supported a causal association between hypertension (OR = 7.203 [95% CI, 2.297-22.587]) and NAFLD. Similar results were observed for the causal exploration between SBP (OR = 1.024 [95% CI, 1.003-1.046]), DBP (OR = 1.047 [95% CI, 1.005-1.090]), and NAFLD. The sensitive analysis further confirmed the robustness and reliability of these findings (all P >0.05). Conclusion::Hypertension was associated with an increased risk of NAFLD.

Objective:To explore the influencing factors of efficacy of combination therapy of nucleos（t）ide analogues（NAs）with pegylated interferon α-2a（Peg IFNα-2a）for patients with chronic hepatitis B（CHB）.Methods:The clinical data of 141 CHB patients receiving combination therapy of NAs and Peg IFNα-2a for ≥48 weeks in the First Affiliated Hospital of Zhejiang University School of Medicine from January 1，2013 to December 1，2023 was retrospectively analyzed. Patients were divided into clinical cure group（ n=45）and non-clinical cure group（ n=96）. Multivariate Logistic regression analysis was used to analyze the influencing factors of clinical cure. Results:Multivariate Logistic regression analysis showed that baseline HBsAg <1 500 IU/mL（ OR=0.160，95% CI 0.049-0.522， P=0.002），a decrease by >1 lg IU/mL in HBsAg at week 48 compared to the baseline level（ OR=0.114，95% CI 0.027-0.484， P=0.003），and sequential combination therapy（ OR=0.351，95% CI 0.128-0.960， P=0.041）were independent predicting factors for the efficacy of combination therapy of NAs and Peg IFNα-2a in CHB patients. Conclusions:The study indicates that the efficacy of the sequential combination therapy is superior to that of the treatment-na?ve therapy，and patients with baseline HBsAg<1 500 IU/mL is more likely to achieve clinical cure than those with higher baseline levels. A primary course of 48 weeks was recommended，with the possibility of an extension of the course by evaluating the degree of HBsAg decline at 48 weeks.

OBJECTIVETo study the severe acute respiratory syndrome (SARS)-associated coronavirus genotype and its characteristics.

METHODSA SARS-associated coronavirus isolate named ZJ01 was obtained from throat swab samples taken from a patient in Hangzhou, Zhejing province. The complete genome sequence of ZJ01 consisted of 29,715 bp (GenBank accession: AY297028, version: gi: 30910859). Seventeen SARS-associated coronavirus genome sequences in GenBank were compared to analyze the common sequence variations and the probability of co-occurrence of multiple polymorphisms or mutations. Phylogenetic analysis of those sequences was done.

RESULTSBy bioinformatics processing and analysis, the 5 loci nucleotides at ZJ01 genome were found being T, T, G, T and T, respectively. Compared with other SARS-associated coronavirus genomes in the GenBank database, an A/G mutation was detected besides the other 4 mutation loci (C:G:C:C/T:T:T:T) involved in this genetic signature. Therefore a new definition was put forward according to the 5 mutation loci. SARS-associated coronavirus strains would be grouped into two genotypes (C:G:A:C:C/T:T:G:T:T), and abbreviated as SARS coronavirus C genotype and T genotype. On the basis of this new definition, the ZJ01 isolate belongs to SARS-associated coronavirus T genotype, first discovered and reported in mainland China. Phylogenetic analysis of the spike protein gene fragments of these SARS-associated coronavirus strains showed that the GZ01 isolate was phylogenetically distinct from other isolates, and compared with groups F1 and F2 of the T genotype, the isolates of BJ01 and CUHK-W1 were more closely related to the GZ01 isolate. It was interesting to find that two (A/G and C/T) of the five mutation loci occurred in the spike protein gene, which caused changes of Asp to Gly and Thr to Ile in the protein, respectively.

CONCLUSIONAttention should be paid to whether these genotype and mutation patterns are related to the virus's biological activities,epidemic characteristics and host clinical symptoms.

Objective To analyze the distribution and antimicrobial resistance profile of clinical bacterial strains isolated from blood culture in China.Methods Clinical bacterial strains isolated from blood culture from participating hospitals of Blood Bacterial Resistance Investigation Collaborative System (BRICS) during January 2014 to December 2015 were collected.Antibiotic susceptibility tests were conducted with agar dilution or broth dilution methods as recommended by US Clinical and Laboratory Standards Institute(CLSI)2018.The data were analyzed with Whonet 5.6 software.Results During the study period,4 801 clinical bacterial isolates were collected from 26 hospitals,of which 1 798 (37.5％) were Gram-positive bacteria and 3 003 (62.5％) were gram-negative bacteria.The top 10 isolates were Escherichia coli (33.8％),coagulase-negative Staphylococcus (19.0％),Klebsiella pneumoniae (11.9％),Staphylococcus aureus (10.1％),Acinetobacter baumannii (4.0％),Pseudomonas aeruginosa (3.8％),Streptococcus (3.0％),Enterobacter sulcus (2.9％),Enterococcus faecium (2.8％) and Enterococcus faecalis (1.8％).Methicillin-resistant Staphylococcus aureus (MRSA) and methicillinresistant coagulase-negative Staphylococcus (MRCNS) accounted for 33.9％ (165/487) and 56.9％ (520/913) of Staphylococcus aureus and coagulase-negative Staphylococcus respectively.No vancomycinresistant Staphylococcus was detected.The resistance rate of Enterococcus faecium to vancomycin was 0.7％ (1/135),and no vancomycin-resistant Enterococcus faecaliss was detected.The positive rates of extendedspectrum β-1actamases(ESBLs)-producing Escherichia coli,Klebsiella pneumoniae and Proteus were 56.9％ (923/1 621),30.1％ (172/572) and 29.2％ (7/24),respectively.The positive rates of carbapenemresistant Escherichia coli,Klebsiella pneumoniae,Enterobacter,Salmonella and Citrobacter were 1.2％ (20/1 621),7.2％ (41/572),4.3％ (6/141),1.5％ (1/67) and 2.9％ (1/34),respectively.The resistance rates of Acinetobacter baumannii to polymyxin and tegacycline were 2.6％ (5/190) and 8.9％ (17/190)respectively,and that of Pseudomonas aeruginosa to polymyxin and fosfomycin were 1.1％ (2/183)and 0.6％ (1/183),respectively.Conclusions The surveillance results from 2014 to 2015 show that the main pathogens of blood stream infection in China are Gram-negative bacteria,while Escherichia coli is the most common pathogen,the detection rate of MRSA is lower than other surveillance data in the same period in China;carbapenem-resistant Klebsiella pneumoniae and Escherichia coli are at a low level as shown in this surveillance.

Objective:To investigate the distribution and antimicrobial resistance profile of clinical bacteria isolated from blood culture in China.Methods:The clinical bacterial strains isolated from blood culture from member hospitals of Blood Bacterial Resistant Investigation Collaborative System (BRICS) were collected during January 2018 to December 2019. Antibiotic susceptibility tests were conducted with agar dilution or broth dilution methods recommended by US Clinical and Laboratory Standards Institute (CLSI). WHONET 5.6 was used to analyze data.Results:During the study period, 14 778 bacterial strains were collected from 50 hospitals, of which 4 117 (27.9%) were Gram-positive bacteria and 10 661(72.1%) were Gram-negative bacteria. The top 10 bacterial species were Escherichia coli (37.2%), Klebsiella pneumoniae (17.0%), Staphylococcus aureus (9.7%), coagulase-negative Staphylococci (8.7%), Pseudomonas aeruginosa (3.7%), Enterococcus faecium (3.4%), Acinetobacter baumannii(3.4%), Enterobacter cloacae (2.9%), Streptococci(2.8%) and Enterococcus faecalis (2.3%). The the prevalence of methicillin-resistant S. aureus (MRSA) and methicillin-resistant coagulase-negative Staphylococcus were 27.4% (394/1 438) and 70.4% (905/1 285), respectively. No glycopeptide-resistant Staphylococcus was detected. More than 95% of S. aureus were sensitive to amikacin, rifampicin and SMZco. The resistance rate of E. faecium to vancomycin was 0.4% (2/504), and no vancomycin-resistant E. faecalis was detected. The ESBLs-producing rates in no carbapenem-resistance E. coli, carbapenem sensitive K. pneumoniae and Proteus were 50.4% (2 731/5 415), 24.6% (493/2001) and 35.2% (31/88), respectively. The prevalence of carbapenem-resistance in E. coli and K. pneumoniae were 1.5% (85/5 500), 20.6% (518/2 519), respectively. 8.3% (27/325) of carbapenem-resistance K. pneumoniae was resistant to ceftazidime/avibactam combination. The resistance rates of A. baumannii to polymyxin and tigecycline were 2.8% (14/501) and 3.4% (17/501) respectively, and that of P. aeruginosa to carbapenem were 18.9% (103/546). Conclusions:The surveillance results from 2018 to 2019 showed that the main pathogens of bloodstream infection in China were gram-negative bacteria, while E. coli was the most common pathogen, and ESBLs-producing strains were in majority; the MRSA incidence is getting lower in China; carbapenem-resistant E. coli keeps at a low level, while carbapenem-resistant K. pneumoniae is on the rise obviously.