With the deepening of population aging， the number of disabled and semi-disabled elderly individuals continues to increase， and the ethical care issues of mental health services for the disabled elderly are becoming increasingly prominent. When examining ethical issues， the concept of the “holistic view of traditional Chinese medicine” from ancient Chinese traditional culture was explored. While applying theories from the holistic view of traditional Chinese medicine， such as “the unity of form and spirit” and “the integration of form and spirit，” emphasis should be placed on the organic integration of psychology and physiology. The mental health status of the disabled elderly can be comprehensively improved by addressing both internal and external environmental factors. In response to the current situation of mental health services for the disabled elderly， ethical care should be provided for the mental health services of the disabled elderly， focusing on the value of subjectivity， difference， and continuity in ethical care， thereby improving the mental health status of the disabled elderly.

Objective:To establish a rapid method to detect the minimum inhibitory concentration (MIC) of imipenem in Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae (KPC-Kp) based on ompK36 gene′s GD mutation. Methods:This was a methodological evaluation study. A total of 258 isolates of Klebsiella pneumoniae were collected from Lishui Municipal Central Hospital from March 2011 to December 2019. Porin gene ompK36 and carbapenemase genes blaKPC, blaNDM, blaIMP and blaOXA-48 were amplified by PCR and confirmed by sequencing. The MIC was detected and confirmed by microbroth dilution susceptibility test, and the corresponding patterns of genotype and MIC were constructed. Based on the patterns, a method for rapid detection of imipenem MIC by real-time fluorescence PCR (RT-PCR) was designed and established. The 159 isolates of non-repetitive Klebsiella pneumoniae collected by Lishui Disease Prevention and Control Center (CDC) from 2017 to 2019 were used for further verification. The sensitivity and specificity were calculated by fourfold table. Kappa test was used to compare the consistency between RT-PCR and microbroth dilution susceptibility test. Results:Among 258 isolates, 109 isolates did not carry carbapenemase gene, 65 isolates carried ompK36 gene GD mutation, 127 isolates carried blaKPC, 15 isolates carried blaNDM, 9 isolates carried blaIMP, and blaOXA-48 was not detected. With mircobroth dilution susceptibility test as the standard, there were 3 corresponding patterns between the drug resistance gene and the imipenem MIC of Kp: when all the 4 carbapenemase genes were negative, MIC≤1 mg/L, the sensitivity was 100% (107/107) and the specificity was 98.4% (125/127); when blaKPC was positive and ompK36 gene GD mutation was negative, 4 mg/L≤MIC≤16 mg/L, the sensitivity was 88.2% (60/68) and the specificity was 98.8% (164/166); when blaKPC and ompK36 gene GD mutation were both positive, MIC≥32 mg/L, the sensitivity was 96.6% (57/59) and the specificity was 96.6% (169/175). RT-PCR detected blaKPC, blaNDM, blaIMP, blaOXA-48 genes accurately.The RT-PCR results of ompK36 gene GD mutation in the KPC-producing isolates were 100% consistent with the sequencing results. In the 159 isolates from Lishui CDC, the sensitivity and specificity of imipenem MIC detected by RT-PCR were higher than 95% in all 3 patterns with mircobroth dilution susceptibility test as the standard, and Kappa value was 0.971. Conclusion:The RT-PCR based on ompK36 gene GD mutation was helpful to quickly determine the MIC range of imipenem in KPC-Kp.

Extracellular vesicles (EVs) are phospholipid bilayer vesicles actively secreted by cells, that contain a variety of functional nucleic acids, proteins, and lipids, and are important mediums of intercellular communication. Based on their natural properties, EVs can not only retain the pharmacological effects of their source cells but also serve as natural delivery carriers. Among them, plant-derived nanovesicles (PNVs) are characterized as natural disease therapeutics with many advantages such as simplicity, safety, eco-friendliness, low cost, and low toxicity due to their abundant resources, large yield, and low risk of immunogenicity in vivo. This review systematically introduces the biogenesis, isolation methods, physical characterization, and components of PNVs, and describes their administration and cellular uptake as therapeutic agents. We highlight the therapeutic potential of PNVs as therapeutic agents and drug delivery carriers, including anti-inflammatory, anticancer, wound healing, regeneration, and antiaging properties as well as their potential use in the treatment of liver disease and COVID-19. Finally, the toxicity and immunogenicity, the current clinical application, and the possible challenges in the future development of PNVs were analyzed. We expect the functions of PNVs to be further explored to promote clinical translation, thereby facilitating the development of a new framework for the treatment of human diseases.

ObjectiveTo explore the mechanism of Shaoyaotang （SYT） in the treatment of ulcerative colitis （UC） based on network pharmacology and experimental verification. MethodThe core components， target genes， and main pathways of SYT were predicted based on network pharmacology， and UC-related components， target genes， and pathways were screened. Dextran sodium sulfate （DSS） was used to induce the UC model in mice， and the effect of SYT on UC mice was observed， followed by mechanism verification. ResultNetwork pharmacology indicated that 174 active components and corresponding 159 target genes of SYT were screened， and the related pathways were those mediated by 5-hydroxytryptamine （5-HT） degredation and 5-HT receptor 3. The results of animal experiments showed that compared with the model group， the SYT group showed increased body weight and colon length（P<0.01）， reduced disease activity index （DAI） score （P<0.01）， improved histopathological manifestations， reduced concentrations of 5-HT in the colonic tissues and serum （P<0.05， P<0.01）， and increased mRNA expression of monoamine oxidase A （MAOA）， monoamine oxidase B （MAOB）， sodium-dependent serotonin transporter （SLC6A4）， and 5-HT receptor 3A （5-HTR3A） related to 5-HT metabolism in the colon （P<0.01）. ConclusionSYT can alleviate the local inflammatory response of the intestinal tract in UC by regulating 5-HT degredation pathways.

Objective:To compare the genetic spectrums of esophageal squamous cell carcinoma (ESCC) patients with different prognosis after postoperative radiotherapy and to screen the genetic variants associated with radiotherapy resistance.Methods:A total of 32 ESCC patients who received radical surgery and postoperative adjuvant radiotherapy in Affiliated Hospital of Nantong University from January 2015 to December 2019 were selected as the study objects. According to whether there was any recurrence in the radiation field within 1 year, they were divided into a recurrence group (radiotherapy resistance group, n=16) and a stable group (radiotherapy sensitive group, n=16). Genomic DNA was extracted from patients and high-throughput sequencing was performed using whole exome sequencing (WES) technology. Biological information analysis software Trimmomatic, BWA and Picard were used to process the data and the alignment files were obtained by GATK comparison, then Vardict software was used to screen out various genetic variants from the sequencing data. The disease free survival (DFS) and overall survival (OS) were estimated by Kaplan-Meier method. Cox proportional hazard regression model was used to analyze the independent risk factors of DFS and OS of ESCC patients. Results:After quality control of the sample data, 26 patients were finally included in this study for follow-up analysis, 13 in each of the recurrence and stable groups. The median tumor mutation burden of non-silent tumors in the whole group was 0.95 mutations/Mb. The substitution types of mutant bases were mainly C>T conversion, followed by C>G transmutation. The genetic variants with the highest frequency were single nucleotide polymorphism (SNP) (75.1%), deletion mutation (13.7%) and insertion mutation (10.5%). The number of tumor-specific mutations in the recurrence group was slightly higher than that in the stable group (median mutation number was 36 and 34, respectively), and the top ten gene profiles of mutation frequency were significantly different between the two groups. In the recurrence group, 392 unique mutated genes were detected, and the top five were MUC19, NPIPA5, EPPK1, FLG and FOXG1. In the stable group, 192 unique mutation genes were detected, and the top five were TCHH, WNK1, AIM1L, COL6A5 and DPCR1. The median DFS and OS were 15.0 months (95% CI: 10.1 months-not reached) and 26.2 months (95% CI: 19.8 months-not reached) in the recurrence group respectively, and no recurrence or metastasis occurred in the stable group. Univariate analysis showed that GRIK2 ( χ2=6.81, P=0.009), MUC4 ( χ2=4.25, P=0.039), MUC5B ( χ2=4.03, P=0.045), PRRG1 ( χ2=5.15, P=0.023) gene mutations, 3p deletion ( χ2=4.16, P=0.041) and 14q deletion ( χ2=7.09, P=0.008) were correlated with DFS. FLG ( χ2=6.41, P=0.011), NPIPA5 ( χ2=4.57, P=0.033), PKD1L2 ( χ2=6.41, P=0.011), FOXG1 ( χ2=4.57, P=0.033) gene mutations, 3p deletion ( χ2=3.88, P=0.049), 14q deletion ( χ2=5.66, P=0.017) and 18p deletion ( χ2=3.85, P=0.050) were associated with OS. Multivariate analysis showed that 14q deletion ( HR=3.65, 95% CI: 1.18-11.32, P=0.025) was an independent risk factor for DFS of ESCC patients with postoperative adjuvant radiotherapy, and FLG ( HR=8.94, 95% CI: 1.52-52.74, P=0.016), NPIPA5 ( HR=6.36, 95% CI: 1.23-33.03, P=0.028) gene mutation and 14q deletion ( HR=3.82, 95% CI: 1.18-12.31, P=0.025) were independent risk factors for OS of ESCC patients with postoperative adjuvant radiotherapy. Conclusion:The WES results suggest that the types and rates of gene mutations of the ESCC patients with postoperative adjuvant radiotherapy in the recurrence and stable groups are basically the same, but the mutation spectrum of the two groups is significantly different. FLG, NPIPA5 gene mutations and 14q deletion can be used as molecular markers to predict the prognosis of ESCC patients treated with postoperative adjuvant radiotherapy.

Objective:To compare the application effect of whole-process seamless nursing and conventional nursing in the perioperative period of ankylosing spondylitis (AS) with kyphosis complicated by cervical spine fracture.Methods:A retrospective cohort study was conducted to analyze the clinical data of 64 patients with AS with kyphosis complicated by cervical spine fracture admitted to Henan Provincial People′s Hospital from April 2017 to December 2022, including 37 males and 27 females, aged 27-73 years [(49.8±14.6)years]. There were 43 patients with vertebral fractures and 21 with intervertebral space fractures. All patients underwent reduction and fixation or correction and fixation. Thirty-two patients admitted from April 2017 to August 2019 received conventional care (conventional nursing group), and 32 patients admitted from September 2019 to December 2022 received whole-process seamless care (seamless nursing group). The two groups were compared concerning the visual analog scale (VAS) before, at 12 hours, 24 hours, 1 month, 3 months after surgery and at the last follow-up, the American Spinal Injury Association (ASIA) score before surgery, at 1 and 3 months after surgery and at the last follow-up, the health survey questionnaire (SF-36) score and patient satisfaction score before surgery and at the last follow-up, as well as the incidence of postoperative complications.Results:All patients were followed up for 6-12 months [(9.8±3.2)months]. There was no statistical difference in preoperative VAS between the two groups ( P>0.05). The values of VAS in the seamless nursing group were (3.9±1.9)points, (4.2±0.7)points, (2.7±0.9)points, (2.6±0.6)points, and (1.7±0.8)points at 12 hours, 24 hours, 1 month, 3 months after surgery and at the last follow-up, respectively, lower than those of the conventional nursing group [(5.7±1.2)points, (5.8±1.1)points, (3.6±1.2)points, (3.2±1.1)points, and (2.4±1.0)points] ( P<0.05 or 0.01). The VAS of the seamless nursing group at 12 hours, 24 hours, 1 month, 3 months after surgery and at the last follow-up was lower than that before surgery (all P<0.05). The VAS at 1 and 3 months after surgery and at the last follow-up was lower than those before and at 12, 24 hours after surgery and the VAS at the last follow-up was lower than those at 1, 3 months after surgery (all P<0.05). The differences among VAS of the conventional nursing group before and at 12, 24 hours after surgery were statistically insignificant (all P>0.05). The VAS of the conventional nursing group at 1 and 3 months after surgery and at the last follow-up was lower than those before surgery and at 12, 24 hours after surgery and the VAS at the last follow-up was lower than those at 1, 3 months after surgery (all P<0.05). There was no statistically significant difference in VAS of the two groups between 12 hours and 24 hours after surgery, and between 1 month and 3 months after surgery (all P>0.05). There was no significant difference in the ASIA scores before surgery between the two groups ( P>0.05). There were significant improvements in ASIA scores in the seamless nursing group at 1, 3 months after surgery and at the last follow-up compared with those of the conventional nursing group ( P<0.05). There were no statistically significant differences in preoperative SF-36 score and patient satisfaction score between the two groups (all P>0.05). At the last follow-up, the SF-36 score and patient satisfaction score of the seamless nursing group were (47.4±6.2)points and (99.5±1.2)points, respectively, which were higher than those of the conventional nursing group [(42.2±7.3)points and (98.1±1.6)points] (all P<0.05). At the last follow-up, the SF-36 score and patient satisfaction score of the seamless nursing group were higher than those before surgery (all P<0.01). The SF-36 score of the conventional nursing group was higher than that before surgery ( P<0.01), but there was no significant difference in patient satisfaction score ( P>0.05). The incidence of postoperative complications in the seamless nursing group was 6.3% (2/32), lower than that of the conventional nursing group [25.0% (8/32)] ( P<0.05). Conclusion:For AS with kyphosis complicated by cervical spine fracture, whole-process seamless nursing is associated with alleviated postoperative pain, improved spinal nervous function, quality of life and degree of satisfaction, and reduced incidence of complications compared with the conventional nursing.

Objective:This study aimed to explore the structural and functional characteristics of the fecal-associated microbiome(FAM)between the phlegm-dampness constitution(PDC)and balanced consti-tution(BC),and to screen the related specific operational taxonomic unit(OTU)biomarkers.Methods:This was a cross-sectional study.After strictly identifying the constitution of subjects,their clinical index was recorded and counted.Fecal samples were collected for 16S rDNA sequencing.Alpha diversity,beta diversity,and the relative abundance of dominant bacterial taxa were used to describe the FAM structure,and the Wilcoxon rank-sum test,MetagenomeSeq,and linear discriminant analysis effect size(LEfSe)were used to screen specific bacterial taxa.Specific OTUs were screened to construct receiver operating characteristic(ROC)curves.Results:Thirty-two subjects were enrolled,including 22 subjects with BC and 10 subjects with PDC.There were significant differences in cold preference,levels of aspartate transaminase,β2-microglobulin,and creatine kinase MB,and alpha diversity indices(Shannon and Shannoneven)between the two groups.In principal coordinate analysis by abund-jaccard distance measure and partial least squares discriminant analysis,bacterial communities clustered separately between the two groups.Furthermore,based on MetagenomeSeq,LEfSe,and the Wilcoxon rank-sum test,a total of 43,18,and 130 OTUs were differentially distributed between BC group and PDC group,respectively,and OTU200,OTU133,and OTU353 were screened when P＜.01.The area under the ROC curve constructed from the 3 selected OTUs was 0.93.Conclusion:The FAM structure and related functional characteristics of the PDC group differed from those of the BC group.In particular,OTU200,OTU133,and OTU353 can be used as unique markers of PDC to assist clinical diagnosis.

目的：探讨环状 RNA circ_0091579 作为分子海绵吸附 miR-330-3p 介导环指蛋白 126（ring finger protein 126， RNF126）对结直肠癌（colorectal cancer，CRC）细胞增殖、凋亡、侵袭的影响。方法：选取 2019 年 2 月至 2020 年 5 月在大理大学 第一附属医院行手术治疗的 60 例 CRC 患者的癌组织和癌旁组织。构建 circ_0091579、miR-330-3p 的过表达或敲减的 CRC LoVo 细胞，qPCR 检测 CRC 组织和细胞中 circ_0091579、miR-330-3p 和 RNF126 的表达；MTT、Transwell、流式细胞术分别检测 细胞的增殖、侵袭、凋亡情况；生物信息学方法预测 circ_0091579 和 miR-330-3p、miR-330-3p 和 RNF126 的靶向关系并用双荧光 素酶报告实验和 RNA 免疫沉淀实验验证。结果：CRC 组织和多种细胞（HCT116、SW620、CW-2、LoVo 细胞）中，circ_0091579 和 RNF126 均高表达、miR-330-3p 均低表达（均 P<0.05）。敲减 circ_0091579 可以抑制 LoVo 细胞的增殖、侵袭而促进其凋亡 （均 P<0.05），但该影响在加入 miR-330-3p 后被逆转；过表达 miR-330-3p 使 LoVo 细胞增殖和侵袭能力减弱但凋亡程度加强 （均 P<0.05），该影响在加入 RNF126 后被抵消。circ_0091579、miR-330-3p 和 RNF126 之间存在靶向作用关系。结论：circ_0091579 通过 miR-330-3p/RNF126 轴促进 LoVo 细胞增殖、迁移和侵袭而抑制其凋亡。

Objective: To understand the attitude of college students in Zhejiang towards e cigarette and its influencing factors, so as to provide basis for making prevention and control strategies. Methods: In September 2020, 10 colleges and universities in Zhejiang Province were selected to conduct an online survey by a combination of typical sampling method and convenience sampling method. A multivariable Logistic regression model was used to analyze the influencing factors. Results: Among 884 subjects, 93 (10.52%) were positive about e-cigarettes, 310 (35.07%) thought e-cigarettes were harmless, 252 (28.51%) thought e-cigarettes were not addictive and 67 (7.58%) of respondents were using e-cigarettes. Multivariate Logistic regression analysis showed that gender, grade, cost of living, whether or not smoking e-cigarettes were harmful to college students attitudes towards e-cigarettes( OR =0.59, 0.47, 1.87, 0.34, 0.54, P <0.05). Conclusion College students in Zhejiang Province have a positive attitude towards electronics and are not active in avoiding the dangers of smoking. To make full use of the work of concentrated trainees in tobacco control, efforts should be made to break the positive image of e-cigarettes. Junior college students should be the prioritized population for intervention, and female students should not be neglected.

OBJECTIVE： To study the improvement effects of Bee venom（BV） plastics on experimental cerebral thrombosis in rats. METHODS： Totally 96 SD rats were randomly divided into sham operation group （normal saline）， model group （plastics blank matrix）， Nimodipine group （positive drug， 4.00 mg/kg） and BV plastics low-dose， medium-dose， high-dose groups （1.67， 3.33， 6.67 mg/kg）， with 16 rats in each group. Rats in sham operation group and Nimodipine group were given medicine intragastrically， while rats in model group and BV plastics groups were given medicine by transdermal smearing. After 5 days of continuous administration， the experimental cerebral thrombosis model was established by ligating the right external carotid artery and pterygomandibular artery， and injecting compound thrombus inducer into the internal carotid artery. The wet mass ratio of right brain to left brain was measured to investigate the degree of brain edema on the infarcted side. The content of Evans blue （EB） in the left and right hemispheres of rats was determined by ultraviolet spectrophotometry to investigate the cerebral vascular permeability. Blood rheology and coagulation function indicators of rats were measured. The pathological changes of brain tissue in rats were observed by HE staining， and the number of survival neuron cells was counted. RESULTS： Compared with the indexes of sham operation group， the cerebral thrombosis model was established successfully. Compared with model group， the area of blue staining in the right brain （infarcted side） of rats in BV plastics groups was significantly reduced， and the right brain/left brain wet mass ratio and the content of EB in the right brain tissue were significantly reduced （P＜0.05 or P＜0.01）. The whole blood viscosity and Casson viscosity of rats in BV plastics groups， and the plasma viscosity of rats in BV plastics medium-dose and high-dose groups decreased significantly （P＜0.01）. PT and APTT of rats were prolonged significantly in BV plastics medium-dose group （P＜0.01）. The pathological changes of brain tissue in rats in BV plastics groups were significantly alleviated. The arrangement of neuron cells was more orderly， the shape and structure of cells were clear， the nucleolus was clear， the membrane was intact， and the number of survival neuron cells was significantly increased （P＜0.01）. CONCLUSIONS： BV plastics can alleviate brain edema， inhibit cerebral vascular permeability， improve hemorheology and coagulation function indicators of rats after the formation of cerebral thrombosis， and alleviate nerve cell injury after ischemia.