The aim of the study is through observing the morphology of the prepared influenza virus (H1N1) with transmission electron microscopy (TEM) and atomic force microscopy (AFM) to explore the application of AFM on the research of the external character of viruses and provide a new, simple and efficient technique for the study of the viral morphology. TEM image was obtained by negatively stained influenza virus with 1% Phosphotungstic Acid; AFM image applied the tapping mode to influenza virus without any further treatment in air at room temperature, and the morphology parameters, including length (diameter), Ra and Rq are calculated by sectional analysis. The shapes of influenza virus A are spherical, filamentous or other pleomorphous particles observed by both AFM and TEM. TEM image of influenza virus A is two-dimensional image, and viral surface has visible spikes, while AFM exhibits the three-dimensional image that can be described with several quantifiable indexes through sectional analysis. AFM phase images show viral surface clearly which is characterized by rugged feature and gear-like protuberance. As compared with TEM, AFM is a new research tool for viral morphology study with the advantages of simple sample preparing, visible interface and is intuitionistic for researchers. The surface characteristic parameters of viruses provided by AFM can be served as the main quantifiable indexes for viral morphological study.
Influenza A Virus, H1N1 Subtype ; ultrastructure ; Microscopy, Atomic Force ; Microscopy, Electron, Transmission

Objective To explore the protective effect of modified Taoren Chengqi Decoction on ventilator-induced lung injury(VILI)in rats based on the nuclear factor-erythroid 2-related factor 2(Nrf2)/glutathione peroxidase 4(GPX4)-ferroptosis pathway.Methods Rats were randomly separated into the control group,the model group,the modified Taoren Chengqi Decoction group,the ML385(Nrf2 inhibitor)group,and the modified Taoren Chengqi Decoction+ML385 group,with 12 rats in each group.The rats in control group underwent tracheal intubation and kept spontaneous breathing.The rats of other groups were subjected to mechanical ventilation for 4 hours.Seven days before mechanical ventilation,medication treatment was carried out once a day for seven days.After mechanical ventilation,ELISA was applied to detect the levels of tumor necrosis factor-α(TNF-α)and interleukin 6(IL-6)in bronchoalveolar lavage fluid(BALF).Lung wet/dry weight ratio and lung tissue pathology of rat were detected.The reagent kit was applied to detect the content of glutathione(GSH),malonaldehyde(MDA),and Fe2+in rat lung tissue.The relative fluorescence intensity of reactive oxygen species(ROS)and 4-hydroxynonenal(4-HNE)in lung tissue was detected by immunofluorescence staining.The mRNA and protein expressions of solute carrier family 7 member 11(SLC7A11),Nrf2,GPX4 were detected.Results Compared with the control group,the lung tissue of rats in model group was severely damaged,the levels of TNF-α and IL-6 in BALF increased,lung wet/dry weight ratio,content of MDA and Fe2+,and the relative fluorescence intensity of ROS and 4-HNE increased,but the content of GSH,the mRNA and protein expressions of Nrf2,SLC7A11,and GPX4 decreased(P＜0.01).Compared with the model group,the pathological damage of lung tissue in the modified Taoren Chengqi Decoction group was improved,the levels of TNF-α and IL-6 in BALF decreased,lung wet/dry weight ratio,content of MDA and Fe2+,the relative fluorescence intensity of ROS and 4-HNE decreased,the content of GSH,the mRNA and protein expressions of Nrf2,SLC7A11,and GPX4 increased(P＜0.01).However,an opposite trend for corresponding indicators in the ML385 group was found(P＜0.01).The pathological injury of lung tissue was alleviated,the levels of TNF-α and IL-6 in BALF decreased,lung wet/dry weight ratio,content of MDA and Fe2+,and the relative fluorescence intensity of ROS and 4-HNE decreased,GSH content,the mRNA expression of Nrf2,SLC7A11 and GPX4,as well as the protein expression of Nrf2 and GPX4 increased in modified Taoren Chengqi Decoction+ML385 group(P＜0.01,P＜0.05).Compared with ML385 group,the pathological injury of lung tissue was alleviated,the levels of TNF-α and IL-6 in BALF decreased,lung wet/dry weight ratio,the content of MDA and Fe2+,and the relative fluorescence intensity of ROS and 4-HNE decreased,GSH content,the mRNA and protein expression of Nrf2,SLC7A11 and GPX4 increased in modified Taoren Chengqi Decoction+ML385 group(P＜0.01).Compared with the modified Taoren Chengqi Decoction group,the pathological damage in lung tissue of rats was intensified,the levels of TNF-α and IL-6 in BALF increased,lung wet/dry weight ratio,the content of MDA and Fe2+,the relative fluorescence intensity of ROS and 4-HNE increased,the content of GSH,the mRNA and protein expressions of Nrf2,SLC7A11,and GPX4 decreased(P＜0.01)in modified Taoren Chengqi Decoction+ML385 group.Conclusion Modified Taoren Chengqi Decoction may improve rat VILI by activating the Nrf2/GPX4 pathway to inhibit ferroptosis.

OBJECTIVETo analyze the genetic differences of Orientia tsutsugamushi (Ot) Sta56 gene between Shandong isolates and other strains deposited in GenBank.

METHODSPCR and restriction fragment length polymorphism (RFLP) were used to amplify the complete sequence of Ot-Sta56 gene. RFLP profiles of Ot were predicted by a computer program according to their complete sequences of Ot-Sta56 gene. PCR amplicon from XDM2 strain was sequenced and analyzed by Clustal X (1.8) and PHYLIP software.

RESULTSThe complete sequences (about 1.6 kbp) of Ot-Sta56 gene were amplified from B16 strain (isolated from patients), FXS2 strain (isolated from A. agrarius) and XDM2 strain. Four species of restriction endonucleases (Hha I, Hinf I, Hae III, Pst I) were used to digest the PCR amplicons from the 3 isolates. When comparing with the RFLP profiles of prototype Ot, the RFLP profiles of PCR amplicons from the 3 isolates were similar to those of Japan Kawasaki strain, but were quite different from the international reference strains Gilliam, Karp, Kato. Results from DNA sequence analysis showed that the complete sequence of Ot-Sta56 gene homology to Japan Kawasaki strain of XDM2 strain was 97%, and deduced amino acid sequence was 92%.

CONCLUSIONData from the complete sequence of Sta56 gene indicated that the genotypes of Ot isolates in Shandong province were similar, but with distinction from the Kawasaki strain.

Amplified Fragment Length Polymorphism Analysis ; Antigens, Bacterial ; genetics ; Bacterial Proteins ; genetics ; Bacterial Typing Techniques ; classification ; DNA, Bacterial ; genetics ; Genes, Bacterial ; Membrane Proteins ; genetics ; Orientia tsutsugamushi ; genetics ; isolation & purification ; Polymorphism, Restriction Fragment Length ; Sequence Analysis, DNA

Salvia miltiorrhiza(Sm) and Salvia castanea f. tomentosa(Sc) hairy roots were used as experimental materials to study the effects of six different carbon sources, galactose, fructose, lactose, glucose, arabinose and sucrose(control), on fresh weight, dry weight, contents and yields of salvianolic acids and tanshinones. The results showed that galactose was most beneficial to the growth of two kinds of hairy roots, while lactose and arabinose were not conducive to their growth. As for Sm hairy roots, fructose significantly promoted the accumulation of salvianolic acid B, and the content increased by 5.801 times and 10.151 times compared with the control group, respectively. Glucose significantly promoted the accumulation of salvianolic acids. The content and yield of rosmarinic acid were 7.674 times and 9.260 times of that of the control group, and the content and yield of salvianolic acid B were 5.532 times and 6.675 times of the control group. For the hairy roots of Sc, galactose significantly increased the content and yield of rosmarinic acid, reaching 7.820 times and 9.944 times of the control group, respectively. Fructose promoted the increase of the content and yield of cryptotanshinone, reaching 9.242 times and 6.609 times of the control group, respectively. The study confirmed the optimal carbon source for the hairy root culture of Sm and Sc, and provided theoretical guidance for large-scale production of Sm drug-derived components and the utilization of Sc.
Carbon ; Plant Roots ; Salvia ; Salvia miltiorrhiza

Child ; Cytomegalovirus Infections ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; Humans

Objective: To analyze and compare therapy responses, outcomes, and incidence of severe hematologic adverse events of flumatinib and imatinib in patients newly diagnosed with chronic phase chronic myeloid leukemia (CML) . Methods: Data of patients with chronic phase CML diagnosed between January 2006 and November 2022 from 76 centers, aged ≥18 years, and received initial flumatinib or imatinib therapy within 6 months after diagnosis in China were retrospectively interrogated. Propensity score matching (PSM) analysis was performed to reduce the bias of the initial TKI selection, and the therapy responses and outcomes of patients receiving initial flumatinib or imatinib therapy were compared. Results: A total of 4 833 adult patients with CML receiving initial imatinib (n=4 380) or flumatinib (n=453) therapy were included in the study. In the imatinib cohort, the median follow-up time was 54 [interquartile range (IQR), 31-85] months, and the 7-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.2%, 88.4%, 78.3%, and 63.0%, respectively. The 7-year FFS, PFS, and OS rates were 71.8%, 93.0%, and 96.9%, respectively. With the median follow-up of 18 (IQR, 13-25) months in the flumatinib cohort, the 2-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.4%, 86.5%, 58.4%, and 46.6%, respectively. The 2-year FFS, PFS, and OS rates were 80.1%, 95.0%, and 99.5%, respectively. The PSM analysis indicated that patients receiving initial flumatinib therapy had significantly higher cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) and higher probabilities of FFS than those receiving the initial imatinib therapy (all P<0.001), whereas the PFS (P=0.230) and OS (P=0.268) were comparable between the two cohorts. The incidence of severe hematologic adverse events (grade≥Ⅲ) was comparable in the two cohorts. Conclusion: Patients receiving initial flumatinib therapy had higher cumulative incidences of therapy responses and higher probability of FFS than those receiving initial imatinib therapy, whereas the incidence of severe hematologic adverse events was comparable between the two cohorts.
Adult ; Humans ; Adolescent ; Imatinib Mesylate/adverse effects* ; Incidence ; Antineoplastic Agents/adverse effects* ; Retrospective Studies ; Pyrimidines/adverse effects* ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy* ; Treatment Outcome ; Benzamides/adverse effects* ; Leukemia, Myeloid, Chronic-Phase/drug therapy* ; Aminopyridines/therapeutic use* ; Protein Kinase Inhibitors/therapeutic use*