OBJECTIVE To construct the simulated traditional Chinese medicine pharmacy based on virtual simulation technology， and assist in the development of the new mode of traditional Chinese medicine dispensing education training. METHODS The field research and questionnaire surveys were conducted to identify the needs of Chinese medicine students and practitioners for the content and presentation of knowledge on the construction of simulated traditional Chinese medicine pharmacy. Taking the laws and regulations on the construction of traditional Chinese medicine pharmacy and the related teaching materials and literature on traditional Chinese medicine preparation as the knowledge source， the virtual simulation technology was applied to build a simulated traditional Chinese medicine pharmacy so as to achieve the functions of browsing the traditional Chinese medicine pharmacy， learning the knowledge of traditional Chinese medicine preparation and practical skills training. A multi-site simulated traditional Chinese medicine pharmacy evaluation scale study was conducted based on platform operational testing. RESULTS A simulated traditional Chinese medicine pharmacy was constructed， consisting of four core modules： video teaching， animation video， simulated pharmacy， and simulated experience. The overall score of evaluation scale was 93.31， with all entries scoring above 80； the ones with evaluation scales above 90 accounted for 92.31% （60/65）. CONCLUSIONS Simulated traditional Chinese medicine pharmacy based on virtual simulation technology meets the learning needs of users and enhances the teaching effect of traditional Chinese medicine dispensing technology training.

bjective　To analyze the drug resistance screening status and drug resistance influencing factors of high-risk groups of drug-resistant pulmonary tuberculosis in Qingdao, and to understand the inclusion of rifampicin patients in treatment, so as to provide a reference for the prevention and treatment of drug-resistant pulmonary tuberculosis. Methods　The medical records of 726 cases of drug-resistant pulmonary tuberculosis among high-risk populations registered in Qingdao from 2018 to 2022 were obtained from the National Health Insurance Information System of the China Center for Disease Control and Prevention. The drug resistance to five anti-tuberculosis drugs, namely isoniazid (INH), rifampicin (RFP), ethambutol (EMB), levofloxacin (Lfx), and amikacin (Am), in the high-risk populations of drug-resistant pulmonary tuberculosis was analyzed. Univariate and multivariate logistic regression were used toidentify factors influencing rifampicin resistance, and the detection and inclusion of treatment for rifampicin-resistant patients were evaluated. Results　Of the 726 subjects, 278 were drug-resistant, with a total drug resistance rate of 38.29%. The drug resistance for the five anti-tuberculosis drugs in descending order was: INH 25.90%(188/726), RFP 22.87%(166/726), Lfx 14.19%(103/726), EMB 11.29%(82/726), Am 2.48%(18/726). Analysis of the drug resistance spectrum showed that among those resistant to one drug, RFP was most common, accounting for 13.67% (38/278); among those resistant to two drugs, INH+RFP was predominant, accounting for 15.83% (44/278); among those resistant to three drugs, INH+RFP+Lfx was most frequent, at 7.19% (22/278); and among those resistant to four drugs, INH+RFP+EMB+Lfx was highest, at 6.12% (17/278). Multivariate logistic regression analysis of rifampicin resistance showed that compared with patients under 25 years of age, the risk of developing rifampicin resistance was lower in the groups aged 45 to under 65 and those aged 65 and above (OR=0.356, 95%CI: 0.181-0.700; OR=0.352, 95%CI: 0.170-0.729). Compared with migrant patients in other provinces, local patients from within the same county or district had a lower risk of developing rifampicin resistance (OR=0.599, 95%CI:0.383-0.962). Compared with patients who were smear-positive at the end of the second month of initial treatment, the risk of developing rifampicin resistance was higher in patients with relapse/return, failure of retreatment/chronic, and other categories of patients (OR=9.380, 95%CI:3.717-23.671;OR=25.749, 95%CI:8.037-82.490; OR=36.651, 95%CI:8.438-159.201). Conclusions　The situation of drug-resistant pulmonary tuberculosis in Qingdao cannot be ignored. Individuals under 25 years old, migrants from other provinces, and patients with relapse/return, failure of retreatment/chronic, and other categories are significant risk factors for developing rifampicin resistance in the high-risk groups of drug-resistant pulmonary tuberculosis.

Osteoarthritis(OA)is a chronic progressive osteoarthropathy in the elderly.Osteoclast activation plays a crucial role in the occurrence of subchondral bone loss in early OA.However,the specific mechanism of osteoclast differentiation in OA remains unclear.In our study,gene expression profiles related to OA disease progression and osteoclast activation were screened from the Gene Expression Omnibus(GEO)repository.GEO2R and Funrich analysis tools were employed to find differentially expressed genes(DEGs).Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses demonstrated that chemical carcinogenesis,reactive oxygen species(ROS),and response to oxidative stress were mainly involved in osteoclast differentiation in OA subchondral bone.Furthermore,fourteen DEGs that are associated with oxidative stress were identified.The first ranked differential gene,heme oxygenase 1(HMOX1),was selected for further validation.Related results showed that osteoclast activation in the pathogenesis of OA subchondral bone is accompanied by the downregulation of HMOX1.Carnosol was revealed to inhibit osteoclastogenesis by targeting HMOX1 and upregulating the expression of antioxidant protein in vitro.Meanwhile,carnosol was found to alleviate the severity of OA by inhibiting the activation of subchondral osteoclasts in vivo.Our research indicated that the activation of osteoclasts due to subchondral bone redox dysplasia may serve as a significant pathway for the advancement of OA.Targeting HMOX1 in subchondral osteoclasts may offer novel insights for the treatment of early OA.

Background and Aims:Sonodynamic therapy(SDT)is an emerging tumor treatment method that primarily utilizes low-intensity ultrasound(US)to activate sonosensitizers,leading to the production of cytotoxic reactive oxygen species(ROS)that kill cells.This study was conducted with an attempt to develop a novel nanoparticle(NPs)formulation containing a sonosensitizer and the small interfering RNA(siRRS1)of tumor-related gene RRS1 to observe its anticancer effects on colon cancer,to provide new strategies for colon cancer treatment. Methods:RRS1 RNA-Seq data were downloaded from the TCGA and GEO databases,and clinical pathological data were collected from 80 patients with colon cancer at Hunan Provincial People's Hospital.The expression of RRS1 in colon cancer and its relationship with prognosis were analyzed.The proliferation and apoptosis of colon cancer cells transfected with siRRS1 were observed.The sonosensitizer tetra(4-carboxyphenyl)porphyrin(TCPP),gene carrier DSPE-PEI,and siRRS1 were assembled into TCPP@DSPE-PEI-siRRS1 NPs(T@D-siRRS1 NPs).After structural characterization and examination of cellular uptake and ROS generation capabilities in vitro,the anticancer effects of T@D-siRRS1 NPs on colon cancer were assessed in cell experiments and tumor-bearing mouse models. Results:Both database analysis and clinical specimen testing indicated that RRS1 expression in colon cancer tissues was higher than that in adjacent normal tissues,and high expression was associated with poor prognosis(HR=1.88,95％CI=1.27-2.79,P=0.002).Univariate and multivariate analyses showed that RRS 1 mRNA expression was an independent risk factor for overall survival in colon cancer patients(HR=1.354,95％CI=1.447-2.215,P=0.031).Knockdown of RRS1 significantly reduced the proliferation capacity of colon cancer cells and increased apoptosis(both P＜0.05).Characterization results showed that the particles were uniform and stable;T@D-siRRS1 NPs entered cells through endocytosis,where TCPP could generate singlet oxygen(1O2)under ultrasound irradiation.Subsequent in vitro and in vivo experimental results demonstrated that transfection with NPs carrying siRRS1 or TCPP(T@D-control siRNA,T@D-siRRS1)combined with US resulted in varying degrees of growth inhibition in colon cancer cells,with the T@D-siRRS1 NPs+US treatment showing the strongest effect(all P＜0.05).Moreover,no significant damage was observed in the vital organs of mice treated with T@D-siRRS1 NPs+US. Conclusion:T@D-siRRS1 NPs plus US integrate SDT and gene therapy,producing an effective synergistic cytotoxic effect on colon cancer cells.Additionally,T@D-siRRS1 NPs demonstrate good safety and biocompatibility,suggesting potential for clinical application.

Objective:To investigate the clinical efficacy of continuous veno-venous hemodia-filtration (CVVHDF) combined with hemoperfusion (HP) HA380 in the treatment of heat stroke patients with multiple organ dysfunction syndrome (MODS).Methods:A retrospective and observational study was conducted. A total of 15 patients with heat stroke combined with MODS who were admitted to the department of intensive care unit (ICU)of Suizhou Central Hospital/Hubei University of Medicine from July to September 2022 were selected as the study objects. All 15 patients were treated with CVVHDF combined with HA380 based on the comprehensive management strategy for severe illness. Organ function indicators [including total bilirubin (TBil), aspartate aminotransferase (AST), creatine kinase (CK), lactate dehydrogenase (LDH), creatinine (Cr), cardiac troponin T (cTnT), myoglobin (Myo), MB isoenzyme of creatine kinase (CK-MB), sequential organ failure assessment (SOFA)] and inflammatory indicators [including white blood cell count (WBC), neutrophil count (NEU), C-reactive protein (CRP), procalcitonin (PCT), and interleukin-6 (IL-6)] were collected. The improvements of the above indexes at admission, after the first HP, after the second HP, after the third HP, and on the 5th day of treatment were compared. Combined with the clinical outcome of patients, the comprehensive efficacy of CVVHDF combined with HA380 in the treatment of severe heat radiation disease was evaluated.Results:There were 10 males and 5 females among the 15 patients. The average age was (64.5±11.5) years old. There were 6 cases of classical heat stroke and 9 cases of exertional heat stroke. Glasgow coma scale (GCS) was 3-8 at admission; SOFA score was 9-17 within 12 hours after admission; acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) was 25-45 within 24 hours after admission. After treatment, the IL-6 level and SOFA score gradually decreased, and there were significant differences in the decrease after the second HP compared to admission [IL-6 (ng/L): 48.37 (15.36, 113.03) vs. 221.90 (85.87, 425.90), SOFA: 8.3±3.3 vs. 11.1±2.4, both P < 0.05]. The PCT level reached its peak after the first HP [12.51 (6.07, 41.65) μg/L], and then gradually decreased, and the difference was statistically significant after the third HP [1.26 (0.82, 5.40) μg/L, P < 0.05]. Compared those at admission, Cr level significantly improved after the first HP (μmol/L: 66.94±25.57 vs. 110.80±31.13, P < 0.01), Myo significantly decreased after the second HP [μg/L: 490.90 (164.98, 768.05) vs. 3?000.00 (293.00, 3?000.00), P < 0.05], After the third HP, the CK level also showed significant improvement [U/L: 476.0 (413.0, 922.0) vs. 2?107.0 (729.0, 2?449.0), P < 0.05]. After CVVHDF combined with 3 times HP treatment, the patient's inflammatory response was gradually controlled and organ function gradually recovered. On the 5th day of the disease course, WBC, PCT and IL-6 levels were significantly improved compared to admission, and AST, CK, LDH, Cr, Myo, CK-MB, and SOFA score were significantly corrected compared with those on admission. The 24-hour survival rate of 15 patients was 86.67%, and the 24-hour, 7-day and 28-day survival rates were both as high as 73.33%. The average mechanical ventilation time of 11 surviving patients was (101.8±22.0) hours, the average continuous renal replacement therapy (CRRT) time was (58.8±11.0) hours, the average length of ICU stay was (6.3±1.0) days, and the average total hospitalization was (14.6±5.2) days. Conclusion:CVVHDF combined with HP HA380 in the treatment of heat stroke patients with MODS can effectively improve organ function and alleviate the inflammatory storm, which is an effective means to improve the rescue rate and reduce the mortality of severe heat stroke patients.

Objective To develop a portable nuclear radiation detector with low-energy γ-nuclide recognition capability for rapid measurement of the dose levels in low-energy radiation fields and identification of nuclides. Methods A digital multi-channel circuit was developed for a detector based on the room temperature semiconductor cadmium zinc telluride, nuclide recognition was achieved using an intelligent nuclide recognition algorithm, and the energy response function G(E) was used to calculate the real-time ambient dose equivalent rate H*(10). Results The portable spectrometer had a minimum detectable energy of 20 keV, and the typical energy resolution for low-energy X-rays was > 4.10% at 59.5 keV and 20℃, enabling accurate identification of ²⁴¹Am nuclide. Conclusion The device has a good measurement performance for low-energy γ/X rays, effectively addressing the limitations of existing devices for monitoring low-energy radiation fields, and provide reliable technical methods for monitoring and emergency response in spent fuel reprocessing plants or nuclear material production plants.

Objective:To investigate the types, incidences, and clinical characteristics of shock in polytrauma patients at different stages after polytrauma.Methods:A retrospective study was conducted on polytrauma patients admitted to multiple trauma centers from June 2020 to December 2021. The inclusion criteria were patients >18 years old and treated due to polytrauma. Exclusion criteria included an admission time of more than 48 h after trauma, a history of malignancy, or metabolic, consumptive, and immunological diseases. The early stage was defined as the period of ≤48 h after polytrauma, and the middle stage was defined as the period between 48 h and 14 days. The patient’s medical history, clinical manifestations, laboratory tests, imaging examination, injury severity score (ISS), and Glasgow coma scale (GCS) were collected. The types, incidences, and clinical characteristics of shock in different stages after polytrauma were analyzed, according to the diagnostic criteria of each type of shock. The differences between the groups were compared by Student’s t test, χ2 test or Mann-Whitney U test. Results:The incidence of the early and middle stage shock after polytrauma were 73.1% and 36.4%, respectively, with statistically significant difference between stages ( P<0.01). There were significant differences in the incidence of hypovolemic shock (83.6% vs. 28.4%), distributed shock (13.7% vs. 80.9%) and cardiogenic shock (3.5% vs. 6.6%) between stages (all P<0.05). The incidence of obstructive shock (8.4% vs. 9.7%, P>0.05) was similar between stages. The incidence of undifferentiated shock was 1.6% and 1.2%, respectively. There were 9.5% patients with multifactorial shock in the early stage and 14.4% in the middle stage. Totally 7 combinations of multifactorial shock were found in different stages after polytrauma. In the early stage, the combination of HS and DS accounted the highest ratio (42.3%) and followed by HS and OS for 28.8%. In the middle stage, the combination of HS and DS was the most common (48.6%) and followed by DS and OS (24.3%). Conclusions:The incidence of shock in polytrauma patients is high. Different types of shock can occur simultaneously or sequentially. Therefore a comprehensive resuscitation strategy is significant to improve the success rate of treatment.

Objective:To investigate the effect of preoperative use of diuretics on cardiac surgery-associated acute kidney injury(CSA-AKI)in elderly patients.Methods:In this single-center retrospective study, 1 638 patients aged ≥60 years and undergone cardiac surgery(including coronary artery bypass grafting, valve replacement and valvuloplasty)in the Department of Cardiovascular Surgery, Linyi People's Hospital between January 2015 and December 2022 were recruited.The last preoperative serum creatinine(SCr)level was taken as the baseline value, and AKI was diagnosed according to the Kidney Disease Improving Global Outcomes(KDIGO)criteria.Patients were divided into an AKI group and a non-AKI group according to whether AKI occurred after surgery.The clinical characteristics of the two groups were compared, and the effect of preoperative use of diuretics on CSA-AKI was evaluated by multivariate Logistic regression analysis.Results:Of 1638 patients enrolled in the study, 284 patients(17.3%)developed CSA-AKI.Compared with the non-AKI group, there were higher proportions of patients in the AKI group receiving furosemide(62.7% or 178/284 vs.46.2% or 626/1 354, χ2=25.397, P<0.001), spironolactone(70.1% or 199/284 vs.49.9% or 676/1 354, χ2=38.284, P<0.001), and hydrochlorothiazide(8.1% or 23/284 vs.3.5% or 47/1354, χ2=12.288, P<0.001). The number of diuretics in the AKI group was higher than in the non-AKI group[2(0, 2) vs.1(0, 2), Z=-6.381, P<0.001], and the proportion of patients using ≥2 diuretics was higher in the AKI group than in the non-AKI group(70.1% or 199/284 vs.49.0% or 664/1354, χ2=41.652, P<0.001). Multivariate Logistic regression analysis showed that, after adjusting for hypertension, diabetes mellitus, hypoalbuminemia, NYHA functional class Ⅲ/Ⅳ, cardiopulmonary bypass during surgery, operative duration≥6 h, postoperative blood transfusion>600 ml, postoperative use of >3 vasoactive drugs and other variables, preoperative use of ≥2 diuretics remained an independent risk factor for CSA-AKI in elderly patients( OR=1.580, 95% CI: 1.042-2.396, P=0.031). Conclusions:AKI is a common complication after cardiac surgery in elderly patients.Preoperative use of ≥2 diuretics used may be an independent risk factor for CSA-AKI.

Objective To observe the expression level of PRMT5 in gastric cancer cells, and explore the effect of knocking down the expression level of PRMT5 on the biological behavior of gastric cancer cells. Methods Western blot was used to detect the expression of PRMT5 protein in gastric cancer cell lines MGC803, SGC7901, MKN45 and human gastric epithelial cells GES-1. siRNA1 and siRNA2 plasmids were transfected to knock down the expression of PRMT5. Cell proliferation and apoptosis assay, transwell assay were used to detect cell proliferation, apoptosis and invasion abilities, respectively. The protein expression levels of β-catenin, cyclin D1 and Bax were detected. Results Compared with GES-1 cells, PRMT5 protein expression levels increased in MGC803, SGC7901 and MKN45 cells (P < 0.001). After knocking down the expression of PRMT5, the cell proliferation and invasion abilities were weakened, the apoptosis rate increased (P < 0.05), the expression of β-catenin and Cyclin D1 protein decreased, and the expression of Bax protein increased (P < 0.001). Conclusion The expression level of PRMT5 is increased in gastric cancer cells. Knockdown of PRMT5 expression level could inhibit cell proliferation, invasion and promote cell apoptosis via reducing Wnt/β-catenin signaling pathway.

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) system is a hotspot of gene editing and gene expression research, in which CRISPR/Cas13 system provides a new direction for RNA interference and editing. In this study, we designed and synthesized the corresponding gRNAs of CRISPR/Cas13a and CRISPR/Cas13b systems in non-homologous end joining (NHEJ) pathway, such as Ku70 and Lig4, and then detected the expression of ku70 and lig4 in HEK293T cells. The CRISPR/Cas13a system could efficiently knockdown the mRNA expression of ku70 and lig4 more than 50%, and CRISPR/Cas13b system also suppressed ku70 and lig4 about 92% and 76%, respectively. Also, CRISPR/Cas13a, b systems could down-regulate Ku70 and Lig4 proteins level to 68% and 53%, respectively. The study demonstrates that the CRISPR/Cas13 system could effectively knockdown the expression of RNA and protein in HEK293T cells, providing a new strategy for gene function and regulation research.
CRISPR-Cas Systems ; DNA Ligase ATP ; genetics ; Gene Expression Regulation ; genetics ; Gene Knockdown Techniques ; HEK293 Cells ; Humans ; Ku Autoantigen ; genetics