‍Hepatitis E is a type of viral hepatitis caused by hepatitis E virus （HEV） infection. There are four main genotypes of HEV， namely genotypes 1‍ — ‍4. Compared with hepatitis B and hepatitis C， previous research on hepatitis E has made slow progress； however， in recent years， hepatitis E has gradually attracted more attention in China， achieving the transition from behind the scenes to the front stage. First， China has a large population base， a large number of people infected with hepatitis E， and a heavy disease burden， and thus it is necessary to attach great importance to the prevention， diagnosis， and treatment of hepatitis E. Second， the World Health Organization （WHO） has put forward the ambitious goal of eliminating viral hepatitis by 2030， and China has made remarkable achievements in the prevention and treatment of hepatitis B and hepatitis C. At present， the incidence and mortality rates of hepatitis E in China have exceeded those of hepatitis A， and the prevention and treatment of hepatitis E in China is of great significance to actively respond to the call of the WHO. Finally， Chinese researchers in the field of hepatitis E have formed Chinese Consortium for the Study of Hepatitis E， formulated related expert consensus， and established international cooperation platforms， which further promotes the research on hepatitis E in China.

Accumulating evidence has confirmed the links between transfer RNA (tRNA) modifications and tumor progression. The present study is the first to explore the role of tRNA methyltransferase 5 (TRMT5), which catalyzes the m1G37 modification of mitochondrial tRNAs in hepatocellular carcinoma (HCC) progression. Here, based on bioinformatics and clinical analyses, we identified that TRMT5 expression was upregulated in HCC, which correlated with poor prognosis. Silencing TRMT5 attenuated HCC proliferation and metastasis both in vivo and in vitro, which may be partially explained by declined extracellular acidification rate (ECAR) and oxygen consumption rate (OCR). Mechanistically, we discovered that knockdown of TRMT5 inactivated the hypoxia-inducible factor-1 (HIF-1) signaling pathway by preventing HIF-1α stability through the enhancement of cellular oxygen content. Moreover, our data indicated that inhibition of TRMT5 sensitized HCC to doxorubicin by adjusting HIF-‍1α. In conclusion, our study revealed that targeting TRMT5 could inhibit HCC progression and increase the susceptibility of tumor cells to chemotherapy drugs. Thus, TRMT5 might be a carcinogenesis candidate gene that could serve as a potential target for HCC therapy.
Humans ; Carcinoma, Hepatocellular/pathology* ; Cell Hypoxia ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Liver Neoplasms/pathology* ; Signal Transduction/genetics* ; tRNA Methyltransferases/metabolism*

Objective:To investigate the prognostic value of serum free triiodothyronine (FT3) in patients with hepatitis E-related acute liver failure (HEV-ALF).Methods:Clinical data of 88 patients with HEV-ALF and 86 patients with acute hepatitis E (AHE) were collected from the member hospitals of Chinese Consortium for the Study of Hepatitis E between January 2016 and December 2021; the data of 100 health subjects who underwent health check-up in Suzhou Municipal Hospital were also collected as healthy control (HC) group. Serum FT3 levels were analyzed in all subjects. HEV-ALF patients were divided into survival group ( n=73) and death group ( n=15) according to their 30 day survival. Correlation between serum FT3 level and prognosis of HEV-ALF patients were analyzed by Cox regression and orthogonal partial least squares discriminant analysis (OPLS-DA). The receiver operating characteristic (ROC) curve and decision curve analysis (DCA) were used to assess the predictive value of serum FT3 levels for predicting the prognosis of patients, and its prediction efficacy was compared with conventional Model for End-Stage Liver Disease (MELD), King’s College Hospital criteria (KCH) and Child-Pugh models. Results:The levels of serum FT3 in HEV-ALF patients were significantly lower than those in AHE patients and HC group ( P=0.006 or <0.001). Cox regression analysis showed that international standardized ratio ( HR=17.984, 95% CI 2.804-115.362), hepatic encephalopathy ( HR=12.895, 95% CI 2.386-69.695) and total cholesterol ( HR=2.448, 95% CI 1.108-5.409) were independent risk factors for death in HEV-ALF patients, and serum FT3 level ( HR=0.323, 95% CI 0.119-0.876) was a protective factor. OPLS-DA results showed serum FT3 levels had high predictive value. ROC curve analysis results showed that the area under the curve was 0.828 (95% CI 0.733-0.900, P<0.001), the sensitivity was 80.00%, and the specificity was 78.08%. DCA showed that FT3 has good prediction ability and decision-making level serum FT3 levels in patients with improvement and fluctuation were significantly higher than those in the patients with deterioration ( P<0.05 or <0.01). Conclusion:Serum FT3 levels are closely related to the prognosis of HEV-ALF patients and it may be used as a biomarker for the prognosis of patients with HEV-ALF.

Objective:To compare the diagnostic efficacies of Sonazoid contrast-enhanced ultrasound (CEUS) and contrast-enhanced magnetic resonance imaging (CE-MRI) in the diagnosis of focal liver lesions (FLLs), and to evaluate the clinical value of Sonazoid.Methods:A total of 58 FLLs in 50 patients who underwent Sonazoid-CEUS and CE-MRI examinations from July 2019 to January 2021 in the First Affiliated Hospital of Zhejiang University School of Medicine were enrolled in this study according to the inclusion criteria. The final diagnostic reference standard was decided by surgical pathology or ultrasound-guided biopsy pathology. Sonazoid-CEUS and CE-MRI features of benign and malignant FLLs were analyzed, and the sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic coincidence rate of the two tests were calculated respectively.Results:There was a statistically significant difference between benign and malignant FLLs in the imaging pattern of homogeneous or heterogeneous intratumoral enhancement in the artery-dominant phase and washout images in the late phase( P<0.001).9.8%(4/41) of the malignant lesions did not decrease until the late phase but decreased in the post-vascular phase. The sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic coincidence rate of the two tests were 97.6%, 52.9%, 83.3%, 90.0%, 84.5%(Sonazoid-CEUS) and 85.4%, 64.7%, 85.4%, 64.7%, 79.3%(CE-MRI), the differences of sensitivity and specificity were not statistically significant ( P=0.125, P=0.687). Conclusions:The vascular phase in Sonazoid-CEUS is still an important diagnostic sign of FLLs, and the unique Kupffer phase can provide additional information for the diagnosis. Sonazoid-CEUS has the same important value as CE-MRI in the diagnosis of FLLs.

OBJECTIVE: To analyze the clinical characteristics and pregnancy outcomes of pregnant women complicated with coronavirus disease 2019 (COVID-19). METHODS: The clinical data of 3 pregnant women with COVID-19 admitted to the First Affiliated Hospital of Zhejiang University School of Medicine from January 19 to February 10, 2020 were retrospectively analyzed. RESULTS: There was one case in the first-trimester pregnancy (case 1), one in the second-trimester pregnancy (case 2) and one in third-trimester pregnancy (case 3). Cough, fever, fatigue, lung imaging changes were the main manifestations. The white cell count, lymphocyte percentage had no significantly changes in case 1 and case 3, while the levels of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), IL-6 and IL-10 elevated. The lymphocyte count and lymphocyte percentage decreased and the inflammatory indicators significantly increased in case 2. All patients were treated with antiviral, antitussive, oxygen inhalation; case 3 received glucocorticoids, case 2 with severe illness received glucocorticoids and additionally gamma globulin. All three cases were cured and discharged. Case 1 with early pregnancy chose to terminate pregnancy after discharge; case 2 chose to continue pregnancy without obstetric complications; and case 3 had cesarean section delivery due to abnormal fetal heart monitoring. CONCLUSIONS The report shows that COVID-19 in pregnancy women could be cured with active treatment, and the maternal and fetal outcomes can be satisfactory.
Antiviral Agents ; therapeutic use ; Betacoronavirus ; isolation & purification ; Cesarean Section ; Coronavirus Infections ; complications ; drug therapy ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Oxygen ; therapeutic use ; Pandemics ; Pneumonia, Viral ; complications ; drug therapy ; Pregnancy ; Pregnancy Complications, Infectious ; drug therapy ; pathology ; Pregnancy Outcome ; Retrospective Studies ; Treatment Outcome ; gamma-Globulins ; therapeutic use

Objective:To analyze the clinical features and risk factors of invasive pulmonary aspergillosis (IPA) in patients with chronic obstructive pulmonary disease (COPD).Methods:Ninety COPD patients with IPA admitted in the First Affiliated Hospital, Zhejiang University School of Medicine and Zhejiang Provincial People’s Hospital from January 2015 to September 2019 were enrolled, and 180 COPD patients without IPA admitted in the same period were selected as control group. The clinical data of the patients in both groups were analyzed retrospectively. Chi square test was used to compare the imaging characteristics of patients in two groups, and multivariate conditional Logistic regression analysis was used to explore the risk factors of IPA in COPD patients. Results:Among 90 cases of COPD with IPA, the culture of lower respiratory tract samples identified Aspergillus fumigatus in 78 cases, Aspergillus flavus in 6 cases, Aspergillus fumigatus with Aspergillus flavus in 1 case, Aspergillu sterrus in 1 case, and Aspergillus nigerwere in 1 case; 1 case of Aspergillus mycelium was found by sputum exfoliation cytology and 2 cases were positive for serum galactomannan. Chest CT images showed patchy infiltrating shadow (87.8%), pleural effusion (36.7%), nodules (33.3%), cavity (18.9%), consolidation shadow (17.8%), halo sign (14.4%) and air crescent sign (2.2%). The incidence of patchy infiltrating shadow, consolidation shadow, halo sign and cavity were higher in COPD patients with IPA compared to control group ( P<0.05 or P<0.01). Multivariate conditional Logistic regression analysis showed that prolonged hospital stay ( OR=1.183, 95% CI 1.047-1.336), combination of two antibiotics ( OR=5.391, 95% CI 1.241-23.421), duration of antibiotic treatment ≥14 days ( OR=5.275, 95% CI 1.586-17.541), cumulative use of antibiotics ( OR=2.270, 95% CI 1.406-3.664) were the risk factors of COPD with IPA ( P<0.05 or P<0.01). Conclusion:The risk factors of IPA in COPD patients include long duration of hospital stay, combination of two kinds of antibiotics, more than 14 days of antibiotic treatment, and more varieties of antibiotics. If the above risk factors exist in patients with COPD, etiology and serology examination and dynamic monitoring of chest CT scan should be performed for early diagnosis and improved prognosis.

Objective:To investigate the serotype distribution, drug resistance and the virulence gene profiles of clinical isolates of Salmonella in Zhejiang Province. Methods:A retrospective study of 463 clinical isolates of Salmonella in nine regions of Zhejiang Province from 2010 to 2017 was conducted. Their serotypes were detected using the Kauffmann-White scheme based on serological detection. Antibiotics susceptibility tests were carried out using K-B disk diffusion method. Eleven virulence genes were detected by polymerase chain reaction. Results:A total of 35 serotypes were identified among the 463 strains of Salmonella. The dominant serotypes were Salmonella typhimurium (41.90%(194/463)) and Salmonella enteritis (22.25%(103/463)). The resistance rate of Salmonella to ampicillin was the highest (66.52%(308/463)), followed by ampicillin/sulbactam (46.87%(217/463)), while low resistance rate to piperacillin/tazobactam was observed (3.24%(15/463)). All strains were sensitive to imipenem and meropenem. Finally, 188 strains (40.60%) of multi-drug resistance were found. The virulence genes hilA, ssrB, marT, siiD, sopB and pagN were found in all Salmonella strains. The virulence gene vexA was found only in Salmonella typhi and Salmonella Dublin; virulence gene icmF was mainly found in Salmonella enteritis. The virulence plasmid gene spvB and pefA were mainly found in Salmonella enteritidis and Salmonella typhimurium and invariably appeared in pairs. The virulence gene cdtB was mainly found in Salmonella typhi and Salmonella paratyphi A. Conclusions:Salmonella typhimurium and Salmonella enteritis are the main clinically isolated Salmonella strains in Zhejiang Province. The situation of multi-drug resistance is severe and a variety of virulence genes are highly detected.

Mitochondria, the power house of cells, are important organelles in eukaryotic cells. Having their own unique and complete DNA (mtDNA) and genetic system, mitochondria play an essential role in cellular energy metabolism, intracel?lular signaling and apoptotic pathways, as well as many other biological functions, which are closely related with cellular met?abolic network. A disruption of mitochondrial genes can therefore result in mitochondrial dysfunction and human diseases, thus they have been widely used in molecular biology, development biology, genetics, forensic identification and clinical diag?nosis. Consequently, sequencing mitochondrial genome has shown great significance in mitochondrial structure and function research. In this review, research progress in mitochondrial genome sequencing method is summarized, mainly focusing on Sanger sequencing, long-PCR and next-generation sequencing. Also rolling circle amplification and indirect sequencing of mtDNA are reviewed. The ambiguities caused by numts in indirect sequencing are mentioned and resolved.

To observe the effect of uniform and shift rotation culture on the formation and activity of the alginate-chitosan (AC) microencapsulated HepLL immortalized human hepatocytes and HepG2 cells aggregates.AC microcapsulated HepG2 and HepLL cells were randomly divided into two groups. Each group was divided into 3 subgroups according to uniform and shift rotation culture.The size and number of aggregates were observed and measured under laser confocal microscopy and inverted microscope dynamically. The amount of albumin synthesis was detected by ELISA, the clearance of ammonia was detected by colorimetry, and diazepam conversion function was detected by high performance liquid chromatography (HPLC).On day 6, 8, 10, 12, 14 and 16, the number and size of the aggregates, albumin synthesis, diazepam clearance and ammonium clearance increased significantly in shift rotation culture group than in uniform group (all<0.01). The albumin synthesis, diazepam clearance, and ammonium clearance in the microencapsulated HepLL groups were significantly higher than those of HepG2 cells at any time (all<0.01).Shift rotation culture can significantly promote the formation and increase the activity of AC microencapsulated HepLL and HepG2 aggregates, and HepLL cells may be more suitable for bioartificial liver than HepG2.
Albumins ; biosynthesis ; metabolism ; Alginates ; Ammonia ; metabolism ; Animals ; Cell Aggregation ; physiology ; Cell Culture Techniques ; methods ; Cell Line, Transformed ; physiology ; Chitosan ; Diazepam ; metabolism ; Glucuronic Acid ; Hep G2 Cells ; cytology ; physiology ; Hepatocytes ; cytology ; physiology ; Hexuronic Acids ; Humans ; Liver, Artificial ; Rotation

Objective To investigate the expression profile of mRNAs in brain samples collected from pronuclear transfer (PNT) mice. Methods Female CD-1 mice were superovulated, and zygotes were collected after mating with adult male mice. Zygotes with two pronuclei were selected for pronuclear transfer manipulation, and then the reconstructed zygotes were transferred into the oviduct of pseudopregnant female mice. The infant mice obtained from pronuclear transfer were called PNT group, while the embryoes that were not performed pronuclear transfer was regarded as control group. Total RNA were extracted from brain samples of both PNT and control mice, and cDNA were labeled with fluorescent dye. Genes that were differentially expressed were identified using the Agilent mouse mRNA array. Gene ontology analysis and pathway analysis were also completed. Results Compared with control group, 392 mRNAs were expressed differentially, which showed more than 2.0 times variation and statistical significance, accounting for 1.7% of all mRNAs. Among those 366 mRNAs were up-regulated and 26 mRNAs were down-regulated. Eleven mRNAs came to 4.0 times variation in total. Gene ontology analysis indicated that differentially expressed genes were significantly enriched in alternative mRNA splicing, small GTPase mediated signal transduction, regulation of insulin receptor signaling pathway, hydrolase activity, transmembrane transporter activity and pyrophosphatase activity. Significant enriched pathway terms contained ion channel transport, fatty acid metabolism, butanoate metabolism, triacylglycerol and ketone body metabolism. Conclusion Pronuclear transfer might influence some key metabolism process in mouse brain.