Objective To study the inhibitory effect of Huidu Yinhua powder from the Orthodox Manual of External Medicine on methicillin-resistant Staphylococcus aureus(MRSA),virulence factor α-hemolysin(Hla)activity,and biofilm formation,and to explore the optimal ratios of Huidu Yinhua powder and provide experimental support for its use.Methods The inhibitory effects of Huidu Yinhua powder and the herbs in the formula on USA300 were analyzed by the minimum inhibitory concentration(MIC),minimum bactericidal concentration(MBC),and disk diffusion assay(K-B method).Hemolysis,neutralization,oligomerization,and Western blot assays were used to verify in which form the drug inhibits the activity of virulence factor α-hemolysin(Hla).A biofilm assay was performed to evaluate the inhibitory effect of Huidu Yinhua powder on biofilm.Orthogonal experiments were performed to explore the optimal ratio of Huidu Yinhua powder.Results Huidu Yinhua powder inhibited the MRSA strain with a MIC90 of 64 mg/mL and an MBC of 256 mg/mL with antibacterial circle diameter of(7.50±0.50)mm.Huidu Yinhua powder inhibited Hla activity by inhibiting Hla secretion.The minimum effective concentration(MEC)was 16 mg/mL,and the MEC of biofilm was 8 mg/mL.In Huidu Yinhua powder,honeysuckle and astragalus only affected the hemolytic activity of MRSA and biofilm formation without inhibiting bacterial growth.The hemolytic activity and biofilm of MEC were both 32 mg/mL.Glycyrrhiza had a strong bacterial inhibitory capacity with a MIC90 of 8 mg/mL and biofilm MEC of 1 mg/mL without showing inhibitory hemolytic activity at subinhibitory concentrations.The orthogonal experiment showed that,at a ratio of honeysuckle,astragalus,and glycyrrhiza in Huidu Yinhua powder of 1∶2∶4,the MIC90 was 16 mg/mL,MEC of hemolytic activity was 8 mg/mL and that of biofilm was 4 mg/mL,both of which were the lowest among the nine groups.Conclusions Huidu Yinhua powder affects the hemolytic activity and biofilm formation of MRSA at subinhibitory concentrations with the optimal ratio of honeysuckle,astragalus,and glycyrrhiza being 1∶2∶4.

Objective:To explore the clinical phenotype and genetic characteristics of children with childhood absence epilepsy caused by KMT2E gene variants. Methods:The clinical data of 1 case of KMT2E gene variant-associated childhood absence epilepsy admitted to the Department of Pediatric Neurology of Linyi People′s Hospital in January 2023 were collected and followed up, and the child and her family were genetically examined by using whole-exome sequencing and Sanger sequencing, and the pathogenicity of mutation loci was analyzed. The Online Mendelian Inheritance in Man, Human Gene Mutation Database, PubMed database, China National Knowledge Infrastructure, and Wanfang database were consulted with the search term " KMT2E" to summarize the clinical phenotype and genetics of the children with epilepsy associated with KMT2E gene variant. Results:The child is a female, presented with typical absence seizures at the age of 3 years and 8 months, with normal development, video electroencephalogram showing widespread spikes and slow waves around 3 Hz accompanied by typical absence seizures. Seizures decreased after valproic acid was applied at full dosage, and were controlled after combination with lamotrigine. Her clinical diagnosis of childhood absence epilepsy was made. The results of whole-exome sequencing showed that the child had a de novo frameshift variant c.2404dup (p.Arg802Lysfs *8) in the KMT2E gene (NM_182931.3), which had not yet been reported domestically or internationally. The c.2404dup variant was interpreted as a pathogenic variant (PVS1+PS2_Supporting+PM2_Supporting) according to the American Society of Medical Genetics and Genomics variant classification criteria and guidelines. Her parents, older brother and younger sister did not carry the variant and had a normal clinical phenotype. A total of 22 patients with epilepsy associated with KMT2E gene variants were retrieved (including this case, a total of 23 cases), including 10 females and 13 males. All of them were autosomal dominant inheritance, with 20 minor variations, including 8 frameshift variants, 7 missense variants, 2 splicing variants, 2 nonsense variants, 1 synonymous variant, and the remaining 3 cases had large fragment deletions (including 2 cases of the whole gene). Clinical manifestations mainly included epileptic seizures (5 cases of absence seizures, 7 cases of focal seizures with or without secondary tonic-clonic seizures, 9 cases of tonic-clonic seizures, 1 case of spasm seizures, 1 case of myoclonic seizures, tonic seizures, and atonic seizures, 3 cases of epileptic status, and 5 cases of refractory epilepsy, with the onset age of epilepsy ranging from neonatal to adolescence), mental retardation (21/23 cases, 4 mild, 5 moderate, and 5 severe), peculiar facial features (11/23), and autism (3/23), etc. Conclusion:KMT2E gene variant-associated epilepsy is an autosomal dominant disorder with a wide spectrum of clinical phenotypes, and the novel variant c.2404dup in the KMT2E gene identified in the present study can lead to childhood absence epilepsy, which enriches the spectrum of mutations and clinical phenotype of the KMT2E gene.

The field of two-dimensional (2D) nanomaterial-based cancer immunotherapy combines research from multiple subdisciplines of material science, nano-chemistry, in particular nano-biological interactions, immunology, and medicinal chemistry. Most importantly, the "biological identity" of nanomaterials governed by bio-molecular corona in terms of bimolecular types, relative abundance, and conformation at the nanomaterial surface is now believed to influence blood circulation time, bio-distribution, immune response, cellular uptake, and intracellular trafficking. A better understanding of nano-bio interactions can improve utilization of 2D nano-architectures for cancer immunotherapy and immunotheranostics, allowing them to be adapted or modified to treat other immune dysregulation syndromes including autoimmune diseases or inflammation, infection, tissue regeneration, and transplantation. The manuscript reviews the biological interactions and immunotherapeutic applications of 2D nanomaterials, including understanding their interactions with biological molecules of the immune system, summarizes and prospects the applications of 2D nanomaterials in cancer immunotherapy.

Objective:To propose a strategy for detecting early gastric cancer (EGC) under high-definition gastroscopy.Methods:Data of 469 lesions of EGC or high grade intraepithelial neoplasia (HGIN) confirmed by pathology detected at The Seventh Medical Center of Chinese People′s Liberation Army General Hospital from January 2013 to January 2020 were collected and gastroscopic images were re-interpreted. The Helicobacter pylori ( HP) infection status, lesion location in the area of atrophy or at the cardia, morphological type of lesions, lesions with/without clear or regular boundary, and lesion color were analyzed for morphological characteristics of EGC and HGIN under high-definition gastroscopy. Results:Among the 469 lesions of EGC or HGIN, HP-negative lesions accounted for 2.1% (10/469) and ulcerative lesions for 7.7% (36/469). Among non-ulcerative lesions of suspected HP infection ( n=423), there were 28 lesions in the cardia outside the atrophic area and 82.1% (23/28) were reddish under white light imaging. There were 29 non-cardiac lesions outside the atrophic area and 82.8% (24/29) were white or showed clear border under white light imaging. Inside the atrophic area, there were 73 elevated lesions, 95.9% (70/73) of which had clear border or irregular depression on the top. There were 293 flat/depressed lesions in the atrophic area, and 90.8% (266/293) had irregular border or were brown under narrow band imaging. Conclusion:According to the status of HP infection, the location and morphological category of lesions, above endoscopic features can be used as clues to detect EGC and HGIN.

Objective:To explore the effects and mechanisms of bortezomib on the proliferation and apoptosis of acute T lymphocyte leukemia cell line Jurkat.Methods:MTT assay was used to test the influence of bortezomib on the proliferation of Jurkat cells.Flow cytometry was used to detect the influence of bortezomib on apoptosis of Jurkat cells.Real-time quantitative polymerase reaction(RT-PCR) was used to detect the effects of bortezomib on the expression of Bax, Bcl-2 and Cox-2 genes in Jurkat cells.Results:The inhibition rates of 5ng/mL, 10ng/mL, 20ng/mL and 40ng/mL bortezomib on Jurkat cells at 24h were (13.23±0.71)%, (39.53±0.95)%, (53.07±1.12)%, (60.43±0.75)%, respectively, and the inhibition rates at 48h were (25.20±0.96)%, (52.80±1.30)%, (60.67±0.64)%, (75.10±1.35)%, respectively.The inhibitory rates of proliferation of Jurkat cells at 72h were (38.37±0.93)%, (60.94±0.85)%, (73.83±5.08)%, (88.37±1.55)%, respectively.The inhibitory rates of proliferation of Jurkat cells increased with the increase of drug concentration and the prolongation of action time, and the differences were statistically significant( F=1 602.202, 1 085.089, 181.034, all P<0.05). Bortezomib (5ng/mL, 10ng/mL, 20ng/mL and 40ng/mL) treatment for 24h, 48h and 72h, the apoptosis rate of Jurkat cells increased with the increase of drug concentration and the prolongation of action time, the differences were statistically significant( F=1 288.571, 223.378, 251.175, all P<0.05). The expression of Bax mRNA in Jurkat cells increased with the increase of drug concentration and time( F=258.446, 518.929, 276.764, all P<0.05). The Bcl-2 mRNA and Cox-2 mRNA expression levels decreased with the increase of drug concentration and the prolongation of action time( FBcl-2 mRNA=236.848, 264.849, 343.968, FCox-2 mRNA=679.404, 1288.681, 1541.850, all P<0.05). Conclusion:Bortezomib can inhibit the proliferation and induce apoptosis of Jurkat cells.Bortezomib can increase the expression of Bax mRNA and decrease the expression of Bcl-2 and Cox-2 mRNA, which may be the molecular mechanism of bortezomib to promote apoptosis.

Objective To investigate the status on accuracy of estimation in applying Richmond Agitation-Sedation Scale (RASS) by nurses of Intensive Care Unit (ICU) for sedation of critically ill patients and to explore its influencing factors. Methods From November 2016 to January 2017, we selected 206 ICU patients and 38 ICU nurses of a ClassⅢ Grade A hospital in Beijing by convenience sampling. Research group and ICU nurses respectively assessed patients' agitation-sedation state with the RASS and analyzed its differences as well as influencing factors. Results There were 143 patients assessed by ICU nurses with the RASS in accordance with that in research group with 69.4% for the accuracy rate. Logistic regression analysis showed that the influencing factors of accuracy of estimation in RASS included sedative, score of Glasgow and delirium (OR=0.065, 1.424, 0.201;P＜0.05). Conclusions The accuracy of estimation in applying RASS by ICU nurses is at a middle level. We should pay attention to patients with sedative, delirium and the low score of Glasgow during applying RASS for critically ill patients.

Aim To investigate the effects of aspirin on Epstein-Barr virus (EBV)-transformed human B-lym-phocytes.Methods EBV-transformed human B-lym-phocytes were treated with certain concentrations of as-pirin.Cellular proliferation was analyzed by MTT as-say.Further evaluation of apoptosis of aspirin-treated cells was performed through light-field microscope, transmission electronic microscope(TEM),propidium iodide(PI)staining and flow cytometric analysis and DNA electrophoresis. Finally, immunoblot analysis was used to determine the expression levels of apopto-sis-associated proteins, proteins involved in mTOR pathway and PU.1 -Bim axis.Results Aspirin treat-ment inhibited proliferation of EBV-transformed human B-lymphocytes.We observed that aspirin treatment in-duced apoptosis in EBV-transformed human B-lympho-cytes,resulting in the decreased number and size of cells.Ultramicroscopic structural analysis via TEM in-dicated that aspirin treatment deformed the cellular nu-cleus,and led to peripheral chromatin and cytoplasmic vacuole.PI staining and flow cytometric analysis indi-cated that aspirin increased the permeability of cell membrane and decreased the viability of treated cells. Agarose electrophoresis revealed DNA smear in aspirin-treated cells.Mechanistically,mTOR signaling was in-hibited in aspirin-treated cells,as evidenced by the de-creased phosphorylation of S6K1 and S6 via immunob-lot analysis.Aspirin treatment led to the decrease of hematopoietic transcription factor PU.1 .Consequently, pro-apoptotic Bim, apoptosis-associated proteins caspase-3 and PARP were activated in aspirin-treated cells.Conclusion Aspirin may show anti-lymphoma effects via its inhibition of proliferation and induction of apoptosis of EBV-transformed human B-lymphocytes, in which mTOR signal pathway and PU.1 -Bim axis may be involved.

Objective To explore the changes of mTOR signaling in LPS/D-gal-induced acute hepatitis in mice . Methods Twenty-six healthy adult female ICR mice were divided into two groups:the control group and experimental group, 13 mice in each group .LPS/D-gal was used to induce acute hepatitis in the mice .The survival of mice was moni-tored within 24 hours after LPS/D-gal challenge .At 6 hours after challenge , samples of serum and liver tissue were collect-ed for further analysis.Results Injection of LPS/D-gal resulted in acute death of the mice within 24 hours.At 6 hours post LPS/D-gal injection , the blood levels of ALT and AST were significantly increased .The mRNA expression of inflammatory cytokines Tnfa and Il6 was up-regulated in LPS/D-gal-induced hapatitis , in which DNA fragmentation and activation of caspase-3 were subsequently observed .Immunoblot analysis showed that both mTOR pathway and NF-κB pathway were ac-tivated.Unexpectedly , inhibition of mTOR signaling could neither decrease the apoptosis in the liver nor increase the sur -vival of mice .Conclusions The results of the present study indicate that mTOR signaling may play pleiotropic roles in the pathogenesis of LPS/D-gal-induced hepatitis .

Objective To isolate and identify viruses from fecal samples of tree shrew with diarrhea.Methods Fecal sample supernatant of tree shrew with diarrhea was inoculated to three cell lines ( Vero, LLC-MK2 and KMB17 ) , and the cytopathic effects on the cells were observed.The infectious particles in the culture supernatant were further ana-lyzed by transmission electron microscopy ( TEM) , genomic RNA-PAGE, rotavirus detection kit, amplification of S1 com-plete segment and bioinformatics analysis.Results Constant cytopathic effects were induced in Vero, LLC-MK2 and KMB17 cell lines after three passages of culture.The results from TEM, RNA-PAGE and rotavirus analysis indicated that they belong to reoviruses.Analysis of the S1 segments revealed that the S1 sequence from KMB17 cell culture had the high-est homology with that of prototype isolate T1L (85%nucleotide homology and 90%amino acid homology), therefore this isolate was named as type I reovirus.The other two S1 sequences from LLC-MK2 and Vero cell culture were identical to have 85%nucleotide homology and 92%amino acid homology with the prototype isolate T3D, named as type III reovirus. Phylogenetic analysis indicated that the isolates in this study are evolutionally adapted to tree shrews.Conclusions It is the first report here that 2 genotypes of Tupaia orthoreovirus are isolated and identified from one fecal sample via three cell lines and viral S1-specific primers, which provides useful guidelines for the isolation and identification of other reoviruses from tree shrew or other hosts.

ted in a loss of cell fusion,which suggests the 928 site on G2 is crucial for cell fusion and the fusion peptide is likely on G2.