Acute lung injury （ALI） is one of the most common and critical diseases in clinical practice， with extremely high morbidity and mortality， seriously threatening human life and health. The pathogenesis of ALI is complex， in which the inflammatory response is a key factor. Studies have shown that NOD-like receptor protein 3 （NLRP3） inflammasomes are involved in ALI through mechanisms such as inflammation induction， increased microvascular permeability， recruitment of neutrophils， oxidative stress， and pyroptosis， playing a key role in the occurrence and progression of ALI. Therefore， regulating NLRP3 inflammasomes and inhibiting the release of inflammatory factors can alleviate the damage in ALI. At present， ALI is mainly treated by mechanical ventilation and oxygen therapy， which have problems such as high costs and poor prognosis. In recent years， studies have shown that traditional Chinese medicine （TCM） can reduce the inflammatory response and the occurrence of oxidative stress and pyroptosis by regulating the NLRP3 inflammasome， thus alleviating the damage and decreasing the mortality of ALI. Based on the relevant literature in recent years， this article reviews the research progress in TCM treatment of ALI by regulating NLRP3 inflammasomes， discusses how NLRP3 inflammasomes participate in ALI， and summarizes the active ingredients， extracts， and compound prescriptions of TCM that regulate NLRP3 inflammasomes， aiming to provide new ideas for the clinical treatment of ALI and the development of relevant drugs.

Objective　By exploring the function of sulfakinin (SK) and sulfakinin receptor (SKR) of Aedes aegypti, it laid a certain experimental basis and theoretical basis for the research and development of new insecticides targeting neuropeptides and their receptors. Methods　This study investigated the roles of SK and its receptor gene in Ae. aegypti using bioinformatics analysis and Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)/Cas9 knockout technology. Subsequently, RNA interference technology was employed to suppress the expression of SK or its receptor in adult mosquitoes. Lastly, transcriptome sequencing technology was utilized to identify and analyze differentially expressed genes between the interference group and the control group in order to gain insights into their functions. Results　It was found that there is only one SK receptor in Ae. aegypti. In addition, during the construction of mutant strains of Ae. aegypti SK and its receptor gene, it was found that only 2% of the G0 generation mutant strains mutated to form chimeras, with a large number of male chimeras dying, and only 14% of female chimeras being able to lay eggs, ultimately resulting in no effective G1 generation mutants. Transcriptome data showed, compared to the control group, 181 genes were significantly differentially expressed after interfering with the SK gene, with 62 genes significantly upregulated and 119 genes significantly downregulated. In addition, after interference with the sulfakinin receptor, 110 genes exhibited significant differential expression, including 20 upregulated and 90 downregulated genes. Cross-analysis of the two datasets identified 46 genes with significant expression changes after interference with sulfakinin or its receptor, with only 4 genes upregulated and the remaining 42 genes significantly downregulated, and the differentially expressed genes were mainly enriched in the metabolic pathway, endocrine system, and digestive system. Conclusions　The SK and its receptor gene are highly conserved and may primarily play roles in regulating the energy metabolism and digestion functions in Ae. aegypti, thus playing an important role in regulating insect growth and development.

[Objective] To understand the resistance mechanisms, virulence characteristics, and pathogenicity of hypervirulent Klebsiella pneumoniae (hvKp), which causes pyogenic liver abscess (PLA), and to provide related data for clinical treatment of infection caused by this type of bacteria. [Methods] We collected three strains of Klebsiella pneumoniae isolated from the liver abscess fluid of patients with liver abscesses in various departments of The First Affiliated Hospital of Xi’an Jiaotong University. The hypervirulent phenotypes were determined by the wire test, and drug sensitivity was assessed using the VITEK 2 compact automatic microbiological analyzer. Molecular characteristics such as podocarp serotypes, multi-locus sequence typing (MLST), virulence genes, and drug resistance genes were identified through whole-genome sequencing. Additionally, a mouse infection model was established to evaluate pathogenicity. [Results] The isolates were sticky, with mucous thread pulling length >5 mm, all of which exhibited high viscosity phenotypes. Except 146007, which is a multidrug-resistant bacterium, the other two strains had higher antibiotic sensitivity. Whole genome sequencing revealed that the isolates were of high-virulence type, carrying the toxin plasmid rmpADC/rmpA2, iron uptake system, bacterial hairs, secretion system, and other virulence factors. All the three isolates tested positive for rmpA/rmpA2 combined with iucA/iutA, indicating they could be classified as hvKp. Multiple resistance genes were detected, such as β-lactamase like blaTEM, blaSHV, and blaCTX-M. The isolates were highly pathogenic to mice, as evidenced by the diseased organs of the deceased mice observed on autopsy. [Conclusion] The three strains of hvKp isolated in this study carried multiple drug-resistant genes and were highly pathogenic, which suggests that in laboratories and clinical practices it is imperative to promptly identify and adopt necessary disposal strategies to prevent further spread of hvKp.

Objective To develop a basic competency assessment scale for generalist nurse in palliative care and test its reliability and validity.Methods The pre-test scale items pool was formed after literature review,internal discussion in the group,consultation with experts and small sample pre-survey. From January to March 2021,the pre-test scale was used to investigate 1000 nurses in various wards including internal medicine,surgery,emergency,obstetrics and gynaecology,and intensive care units.The results were analyzed by SPSS software and AMOS software to undertake item analysis,exploratory factor analysis and confirmatory factor analysis to develop the final scale.Results Five common factors and 37 entries were identified by factor analysis and explained 71.031％ of the total variance.The Cronbach's α coefficient of the scale was 0.953,the split-half reliability and the test-retest reliability of the scale was 0.772 and 0.963.The confirmatory factor analysis model fitted well,with x2/df=2.57,root mean square error of approximation (RMSEA)=0.059,comparative fit index (CFI)=0.909,incremental fit index(IFI)=0.910,and Tucker-Lewis index(TLI)=0.901.Conclusion Our developed basic competency assessment scale for generalist nurse in palliative care has good reliability and validity,and can be used to evaluate the basic competency of palliative care for generalist nurses.

Objective:To investigate the correlation between Piwi-interacting RNA (piRNA) and diabetic nephropathy (DN).Methods:The differential expression profiles of piRNAs in renal tissues of patients with DN (experimental group) and renal tissues adjacent to tumors of patients with renal tumors (control group) were detected by high-throughput sequencing. The biological function of differentially expressed piRNAs was described by gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis. Real-time fluorescence quantitative PCR was used to detect the serum expression level of target piRNAs in patients with DN. Spearman correlation analysis was used to analyze the correlation between serum target piRNAs and clinical indexes of patients with DN.Results:The results of high throughput sequencing showed that there were 127 differentially expressed piRNAs between DN group and control group, with screening condition of |log 2(fold changes)|≥2 and P<0.05. Among them, there were 99 up-regulated piRNAs and 28 down-regulated piRNAs. The top 5 up-regulated piRNAs were piRNA-hsa-161686, piRNA-hsa-349255, piRNA-hsa-355720, piRNA-hsa-151229 and piRNA-hsa-154959, respectively. The top 5 down-regulated piRNAs were piRNA-hsa-1929960, piRNA-hsa-174194, piRNA-hsa- 148658, piRNA-hsa-172594 and piRNA-hsa-172421, respectively. The PCR verification results of 3 up-regulated genes and 3 down-regulated genes with low P values and high expression levels showed that serum expression level of piRNA-hsa-77976 was significantly down-regulated in patients with DN ( P=0.028), which was consistent with that of sequencing, while the expression levels of other genes were inconsistent with the sequencing results or had no statistical significance. Bioinformatics analysis results predicted that significantly differentially expressed piRNAs might participate in the regulation of DN through Rap1, Ras, PI3K-Akt and axon guiding pathways. The results of correlation analysis showed that the expression level of piRNA-hsa-77976 was negatively correlated with blood urea nitrogen ( r=-0.584, P=0.028), serum creatinine ( r=-0.637, P=0.014), cystatin C ( r=-0.738, P=0.003) and β2 microglobulin ( r=-0.822, P<0.001), and positively correlated with estimated glomerular filtration rate ( r=0.661, P=0.010). Conclusion:The differential expression of piRNA is closely related to DN, and may be used as a new biomarker for the diagnosis and prognosis of DN.

Objective: To construct the full-length prokaryotic expression plasmid of the wild type of androgen receptor (AR) and the truncated body of four functional domains，and to identify the fusion protein by Western blot and electrophoretic mobility shift assay ( EMSA) ． Methods: Based on the pGEX-4T-1 vector ，the recombinant plasmids were constructed to express the full-length and functional domains of AR. IPTG was used to induce the expression of the recombinant proteins，which were isolated and purified by glutathione sepharose 4B beads under the optimized condition．The specific protein expression in the bacterial lysate and the purified protein isolated with glutathione sepharose 4B beads was identified by Western blot with AR antibody and GST labeled antibody．The purified protein was incubated with a fluorescent probe of the virus，and the complex was detected by electrophoresis in a non-denaturing gel. Results : The prokaryotic recombinant plasmids of full length and three functional domain truncated AR were successfully constructed．The recombinant clones were identified by using bacterial culture as a template，and further verified by double enzyme digestion．It showed that there were identical bands in the same sizes as the inserted fragments．The nucleotide and the amino acid sequences were aligned to the reference sequence in NCBI GenBank．The GST fusion protein，GST-AR-NTD + DBD (96 ku) and GST-AR-NTD (86 ku) were successfully induced and verified． The purified protein could be directly combined with the viral genome DNA． Conclusion The prokaryotic expression conditions of truncated AR plasmid from the same gene sequence are different．The purified AR protein can be used to understand the direct interaction mechanism between functional domains of AR and other molecules.

Objective To explore the risk of venous thromboembolism (VTE), especially lower-extremity deep vein thrombosis (DVT) and pulmonary embolism (PE), for stroke patients in rehabilitating, and the functional outcome. Methods A total of 3 557 stroke patients in the neurological rehabilitation center of Beijing Bo'ai Hospital for stroke rehabilitation from January, 2015 to October, 2020 were reviewed through the electronic medical record system. Demographic characteristics, stroke characteristics (type and location), laboratory data (D-dimer polymer and arterial partial pressure of oxygen), motor function (Brunnstrom stage, Fugl-Meyer Assessment of motor and balance, modified Ashworth Scale score of triceps crus, and Holden Walking Ability Classification), activities of daily living (Barthel Index), and anticoagulant/antiplatelet treatment data were collected and analyzed. Results The incidence of DVT and PE was 28.5% and 1.29%, respectively. Most were found 30 days later after onset. The incidence of PE was higher after ischemic stroke (χ2 = 12.49, P < 0.001) rather than hemorrhagic stroke. The patients with hemispheric stroke, severe lower-extremity paralysis, and poor activities of daily living were more prone to complications associated with VTE. After rehabilitation, the function of stroke patients with PE could be improved (|t| > 4.302, P < 0.001). Conclusion The risk of DVT and PE in patients during stroke convalescence may not be negligible, and those with older age, previous history of thrombosis, severe stroke, and severe limb paralysis may be stratified in high-risk. Following anticoagulation treatment, early individualized comprehensive rehabilitation can be done for patients with PE to improve their function and activities of daily living.

Objective To investigate the incidence of Holmes tremor (HT) after stroke and its outcome after medication and rehabilitation. Methods Patients diagnosed as HT after stroke in the ward of neurorehabilitation department from October, 2019 to September, 2021 were reviewed the clinical features, imaging manifestations, drug treatment plan, rehabilitation evaluation scales scores, rehabilitation plan and outcome. Results There were five inpatients with HT (0.7%, 5/715), and all were hemorrhagic stroke, accounting for 1.7% of hemorrhagic stroke. The lesions were located in the midbrain and pons in three cases, cerebellum in one case and thalamus in one case. The tremor appeared 1.5 to seven months after stroke, limited on head and limbs, with other neurological dysfunction. After the comprehensive treatment of drugs and rehabilitation, tremor improved in four cases, and ineffective in one case. The motor and balance function improved less, and the activities of daily living improved somehow. Conclusion The incidence of Holmes tremor is low in stroke patients. The tremor might respond to the treatment, but motor function would not.

Objective : To detect the changes of autophagic flux at different stages after oxygen⁃glucose deprivationreoxygenation (OGD/R) with several highly sensitive methods. Methods : Primary cortical neurons after oxygen deprivation of sugar after reoxygenation (OGD/R) were divided into the experimental OGD/R group and OGD/R + bafilomycinA1 (BafA1) group , using an RFP⁃GFP series fluorescent tags LC3 gene transfection detection cytolysosome and fusion of lysosomes , transmission electron microscope (TEM) observation the ultrastructure of autophagy , p62/SQSTM1 combining LC3 protein to flip the experimental testing p62 and LC3 protein quantitative , p62 immune staining observing the distribution and content. Results : Under fluorescence microscope , the ratio of autophagy lysosome to autophagosome increased significantly in OGD/R group , and the changes of autophagy structure in different stages could be observed in TEM. The ratio of soluble p62 and LC3 Ⅱ/ Ⅰ reflected the activation of autophagic flux , and p62 was mainly distributed in BafA1 group after fluorescence staining. Conclusion Each method has its own advantages , and different methods and indicators can accurately reflect the specific changes of autophagic flux in different stages after neuronal OGD/R. Mastering and applying these methods can effectively explore central nervous system diseases from the perspective of autophagy.

Objective:To investigate the therapeutic efficacy of chimeric antigen receptor T cell (CAR-T) for treatment of relapsed patients with double expression lymphoma after autologous hematopoietic stem cell transplantation (auto-HSCT).Methods:The treatment process of one patient with double expression diffuse large B-cell lymphoma who received CAR-T immunotherapy after the recurrence of auto-HSCT in Henan Provincial People's Hospital in August, 2017 was retrospectively analyzed, and the related literature was reviewed.Results:A 50-year-old female double expression diffuse large B-cell lymphoma patient received the standard treatment regimen, and then had auto-HSCT based on the BEAM preconditioning regimen. The patient relapsed after 5 months, and finally got sustained remission after chemotherapy regimen containing cladribine for 2 courses of treatment combined with CAR-T therapy. CD20 +-CAR-T cells were detected in this patient for 8 mouths sustainably. Conclusion:For relapsed patients with non-Hodgkin lymphoma after auto-HSCT, reduction remission of the treatment regimen containing cladribine followed by CAR-T sequential regimen may be a better treatment option.