Objective:To study the expression of stomatin-like protein 2(STOML2)in oral squamous cell carcinoma(OSCC)tissue and the effects of STOML2 on the tumorigenicity of OSCC cells(OSCCCs)in vitro and in vivo,and the related mechanism.Methods:The protein expression of STOML2 in OSCC and adjacent tissues of 56 patients was detected.OSCCCs SCC-15 were divided into 2 groups.Stom12-siRNA plasmid was transfected into the cells of experimental group and Mock-siRNA plasmid was transfected into the cells of control group.The mRNA and protein expression of STOML2,CDK4 and P16 in the cells was detected by qPCR and Western blot respectively.The cell cycle of the cells was detected by flow cytometry,and the proliferation of the cells was detected by CCK8 asay.The tumorigenicity of the cells was detected by subcutaneous tumor model in nude mice.Results:The positive rate of STOML2 in OSCC and adjacent tissues was 92.86％(52/56)and 8.93％(5/56)respectively(P＜0.001).After siRNA transfection,STOML2 mRNA expression in SCC-15 cells of experimental group and control group was(0.43±0.09)and(1.23±0.19),STOML2 protein ex-pression was(0.52±0.11)and(0.94±0.17)respectively.CDK4 expression was(0.33±0.13)and(1.18±0.17),P16 expression was(0.93±0.12)and(0.29±0.03),respectively.In CCK8 assay the absorbance of SCC-15 cells in experimental group and control group was(1.11±0.24)and(2.19±0.28),in flow cytometry the percentage of cells in G2/M phase was 35.72％±5.33％and 18.65％±3.71％(P＜0.05),respectively.In vivo test showed that the volume(μm3)of subcutaneous transplanted tumor was 1 192.07 ±250.9 and 2 280.5±600.1,the weight(g)of mice was 0.65±0.30 and 1.62±0.40,respectively.Conclusion:STOML2 expression increases in OSCC,STOML2 affects the tumorigenic ability of OSCCCs in vitro and in vivo by regulating P16 related pathways.

CyberKnife has the characteristics of good repeatability of radiotherapy target,high precision and less trauma.It is a stereotactic body radiation therapy system for real-time tracking of moving tumors during treatment.For intrahepatic tumors being highly affected by respiratory movements in treatment,CyberKnife provides precise positioning,planning,and treatment.This paper focuses on the treatment of hepatocellular carcinoma(HCC)using CyberKnife,and reviews as following:the structure and iteration of CyberKnife;image-tracking modes and their advantages and disadvantages;tracking modes of common fiducial markers and their optimization methods,the positioning problems after the implantation of fiducial markers.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Abnormally activated CDK9 participates in the super-enhancer mediated transcription of short-lived proteins required for cancer cell survival. Targeting CDK9 has shown potent anti-tumor activity in clinical trials among different cancers. However, the study and knowledge on drug resistance to CDK9 inhibitors are very limited. In this study, we established an AML cell line with acquired resistance to a highly selective CDK9 inhibitor BAY1251152. Through genomic sequencing, we identified in the kinase domain of CDK9 a mutation L156F, which is also a coding SNP in the CDK9 gene. By knocking in L156F into cancer cells using CRISPR/Cas9, we found that single CDK9 L156F could drive the resistance to CDK9 inhibitors, not only ATP competitive inhibitor but also PROTAC degrader. Mechanistically, CDK9 L156F disrupts the binding with inhibitors due to steric hindrance, further, the mutation affects the thermal stability and catalytic activity of CDK9 protein. To overcome the drug resistance mediated by the CDK9-L156F mutation, we discovered a compound, IHMT-CDK9-36 which showed potent inhibition activity both for CDK9 WT and L156F mutant. Together, we report a novel resistance mechanism for CDK9 inhibitors and provide a novel chemical scaffold for the future development of CDK9 inhibitors.

Abstract Child abuse is a global public health problem, which has emerged as a neglected yet pressing issue in global development. Early and accurate identification of abuse at a lower-age group is of great significance for treatment, which might reduce the risk of re-maltreatment and promote children s physical and mental health development. Therefore, by reviewing the clinical characteristics, risk factors and existing abuse identification and screening tools of child abuse, the study aims at providing basic evidence for the development of child abuse risk identification tools and the establishment of child maltreatment system in China, so as to take timely intervention measures to prevent adverse outcomes or reduce their severity.

Objective:To explore the status quo of the readiness of evidence-based nursing practice for the maintenance and removal of children's central venous catheter (CVC) in 6 domestic medical institutions Pediatric Intensive Care Unit (PICU), analyze the obstacles faced by the application of CVC maintenance and removal evidence in PICU, so as to provide basis for formulating reform strategies.Methods:This study was a cross-sectional survey. In December 2019, 169 nurses from PICU of 6 medical institutions in Shanghai, Guangzhou, Shenzhen, Hangzhou, Xiamen and Hefei were selected by convenience sampling for investigation. The General Information Questionnaire and Clinic Readiness to Evidence-based Nursing Assessment (CREBNA) were used to evaluate the readiness of clinical nurses to evidence, organizational environment and promoting factors in the process of evidence-based practice. A total of 169 questionnaires were issued, and 169 valid questionnaires were recovered, with a valid recovery rate of 100%.Results:A total of 169 nurses were investigated in 6 evidence application sites. The total score of the multi center CREBNA was (136.96±15.33), which was 88.36% of the full score. The scores of the three dimensions were as follows: organizational environment (40.31±4.45), evidence dimension (53.43±6.14), and promoting factors (43.22±5.81). Four of the last five items in the score ranking were from the promoting factor, and the low score items in four places were all from the promoting factor.Conclusions:The best practice project of children's CVC maintenance in PICU is feasible. However, there are still some deficiencies in the preparation of various evidence application sites, especially in the promoting factors, which should be constantly improved to further promote and maintain the implementation of change.

Objective:To investigate frailty status of the elderly patients with leukoaraiosis (LA) and to analyze the correlation between white matter hyperintensities and their frailty status.Methods:From June 2019 to September 2020, a total of 162 patients with leukoaraiosis over 65 years old were screened by cranial MRI.The Fried frailty phenotype was used to evaluate their frailty status.The Fazekas scale scoring method was used to independently assess the periventricular white matter hyperintense (PVH) and deep white matter hyperintense (DWMH) by the cranial MRI images.SPSS 22.0 software was used for statistical analysis.ANOVA test was used to compare normal distribution data between groups, and Kruskal-Wallis H test was used to compare non-normal distribution data between groups.Spearman rank correlation analysis was used to analyze the correlation between PVH and DWMH scores and Fried frailty phenotype score. Results:Among 162 elderly patients with leukoaraiosis, 46 patients (28.40%) were non-frailty, 76 patients (46.91%)were pre-frailty and 40 patients (24.69%) were frailty.There were statistically significant differences in age( F=9.382, P<0.01), number of chronic diseases( H=10.736, P<0.01), number of medication ( H=15.927, P<0.01) and mini-nutritional assessment short form (MNA-SF) scores( F=5.263, P<0.01) among older LA patients with different frailty phenotype.There was statistical difference in PVH scores in elderly LA patients with different frailty phenotype (χ 2=108.537, P<0.01), but no significant difference in DWMH scores (χ 2=4.239, P>0.05). Spearman correlation analysis showed significant positive correlation between PVH score and frailty phenotype score in elderly LA patients ( r=0.718, P<0.001). Conclusion:Elderly LA patients have a high incidence of frailty, which may be related to aging, multi-disease coexistence, multiple medication, nutritional risk and other factors.The occurrence of weakness in elderly LA patients is related to periventricular white matter lesions, and the more serious the white matter damage, the more obvious the degree of frailty.

Whether direct manipulation of Parkinson's disease (PD) risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue. Here, we used an adeno-associated virus serotype 9 (AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras (SNs) of two monkey groups: an old group and a middle-aged group. After the operation, the old group exhibited all the classic PD symptoms, including bradykinesia, tremor, and postural instability, accompanied by key pathological hallmarks of PD, such as severe nigral dopaminergic neuron loss (>64%) and evident α-synuclein pathology in the gene-edited SN. In contrast, the phenotype of their middle-aged counterparts, which also showed clear PD symptoms and pathological hallmarks, were less severe. In addition to the higher final total PD scores and more severe pathological changes, the old group were also more susceptible to gene editing by showing a faster process of PD progression. These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys. Taken together, this system can effectively develop a large number of genetically-edited PD monkeys in a short time (6-10 months), and thus provides a practical transgenic monkey model for future PD studies.
Animals ; Brain ; CRISPR-Cas Systems/genetics* ; Dependovirus/genetics* ; Haplorhini ; Phenotype ; Protein Kinases/genetics*

The TEA domain (TEAD) family proteins (TEAD1‒4) are essential transcription factors that control cell differentiation and organ size in the Hippo pathway. Although the sequences and structures of TEAD family proteins are highly conserved, each TEAD isoform has unique physiological and pathological functions. Therefore, the development and discovery of subtype selective inhibitors for TEAD protein will provide important chemical probes for the TEAD-related function studies in development and diseases. Here, we identified a novel TEAD1/3 covalent inhibitor (DC-TEADin1072) with biochemical IC

Whether direct manipulation of Parkinson’s disease (PD) risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue. Here, we used an adeno-associated virus serotype 9 (AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras (SNs) of two monkey groups: an old group and a middle-aged group. After the operation, the old group exhibited all the classic PD symptoms, including bradykinesia, tremor, and postural instability, accompanied by key pathological hallmarks of PD, such as severe nigral dopaminergic neuron loss (>64%) and evident α-synuclein pathology in the gene-edited SN. In contrast, the phenotype of their middle-aged counterparts, which also showed clear PD symptoms and pathological hallmarks, were less severe. In addition to the higher final total PD scores and more severe pathological changes, the old group were also more susceptible to gene editing by showing a faster process of PD progression. These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys. Taken together, this system can effectively develop a large number of genetically-edited PD monkeys in a short time (6–10 months), and thus provides a practical transgenic monkey model for future PD studies.