This study was aimed at creating an engineered strain of Bacillus subtilis for efficient expression of biologically active type 2 toxin B(TcdB2)derived from a highly virulent strain of Clostridium difficile.The TcdB2 gene was cloned from ST1/RT027 strain genome DNA,incorporated into the PHT01 vector,and then transformed into B.subtilis strain WB800N for prokaryotic expression.Cell toxicity assays revealed that the recombinant TcdB2 exhibited cytotoxic effects in various cells.The engineered B.subtilis strain effectively expressed biologically active TcdB2,thus providing a basis for further exploration of the pathogenic mechanisms of highly virulent strains of C.difficile and establishing a foundation for potential vaccine can-didate targets.

Objective:To analyze the efficiency of human resource allocation and its spatial distribution characteristics of district-level Center for Disease Control and Prevention(CDC)in city N of Jiangsu Province in 2020,in order to provide a strong decision-making reference for optimizing and strengthening the CDC talent team.Methods:The efficiency of human resources of district-level CDC of City N in2020 was measured using the Super-Efficiency SBM model,and the spatial association pattern was analyzed using the natural break point classification method and Moran's index,with the visualization presented through LISA maps.Results:The overall level of human resource efficiency in district-level CDC of City N is relatively high.However,spatially,there are significant differences among the regions,showing a trend of high efficiency in the central areas and low efficiency at the ends.Moran's index and LISA maps indicate a negative spatial correlation in efficiency,with a low-high(L-H)cluster centered on Area L and a high-low(H-L)cluster centered on Area J.The high-high(H-H)cluster pattern has not yet formed,exhibiting a characteristic of interspersed high and low efficiency.Conclusions:There are regional differences in the human resource efficiency of the Disease Control Center in City N,and the spatial cluster pattern needs to be optimized.It is recommended to focus on efficiency improvement in Areas P and L,formulate appropriate policies,and promote coordinated regional development.

Objective:To compare the precision and stability of handheld full-course visual navigation and robot-assisted total hip arthroplasty (THA), as well as to evaluate perioperative laboratory indicators and clinical efficacy.Methods:A retrospective analysis was performed on 68 patients with unilateral hip joint disorders who underwent primary THA at the General Hospital of the People's Liberation Army of China from January to June 2024. Patients were divided into two groups based on the assistance method: the Visual Treatment Solution (VTS) group, which included 34 patients (13 males, 21 females; mean age 56.92±8.31 years; body mass index[BMI] 24.20±3.55 kg/m 2), and the MAKO group, also with 34 patients (18 males, 16 females; mean age 57.97±6.54 years; BMI 25.20±3.91 kg/m 2). Among the VTS group, 1 there were 18 cases on the left side and 16 cases on the right side, including 14 cases of femoral head necrosis, 8 cases of congenital hip dysplasia, and 12 cases of hip osteoarthritis. In the MAKO group, there were 19 cases on the left side and 15 cases on the right side, including 13 cases of femoral head necrosis, 10 cases of congenital hip dysplasia, and 11 cases of hip osteoarthritis. Preoperative planning, intraoperative verification deviations, postoperative acetabular prosthesis anteversion and abduction angles, and limb length discrepancies were analyzed. Acetabular angle distribution scatter plots were generated for intraoperative verification and postoperative imaging. The operation time, hospital stay, and laboratory values, including hemoglobin (Hb) loss, fibrinogen degradation products (FDP), D-dimer, high-sensitivity C-reactive protein (hCRP), as well as postoperative pain (measured by the visual analogue scale, VAS), hip Harris score (HHS), and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores were recorded. Results:No statistically significant differences were observed between postoperative imaging measurements of anteversion and abduction angles and intraoperative verification values ( P>0.05). Postoperative imaging indicated that 91% (31/34) of the VTS group and 100% (34/34) of the MAKO group were within the Lewinnek safety zone. The scatter distribution of angles in the MAKO group was more concentrated than that in the VTS group. Furthermore, the angle deviation between intraoperative verification and preoperative planning was significantly greater in the VTS group for both anteversion (2.00° [-1.00°, 4.00°] vs. 0.00° [-1.00°, 1.00°]) and abduction angles (-1.00° [-5.00°, 0.00°] vs. 0.00° [-1.00°, 1.00°]; P<0.05). The bilateral limb length discrepancies were 1.35(0.67, 2.45) mm in the VTS group and 1.65(0.50, 2.52) mm in the MAKO group, with no significant difference noted ( P>0.05). Both methods achieved effective hip biomechanical reconstruction. Additionally, no significant differences in operation time, Hb loss, FDP, D-dimer, hCRP, or postoperative hospital stay were found ( P>0.05). Follow-up at one and three months postoperatively showed no significant differences in VAS (2.94±0.92 vs. 2.97±0.83), HHS (69.88±4.20 vs. 70.06±3.88), or WOMAC scores (22.44±3.15 vs. 22.76±3.39) between the two groups ( P>0.05). Conclusion:The stability of preoperative planning and design in robot-assisted total hip arthroplasty is superior to that of handheld navigation methods. Nevertheless, both approaches demonstrate comparable postoperative prosthesis stability, laboratory indicators, and clinical outcomes.

Oxidative stress(OS)is a major reason for body damage. Studies have shown that a variety of factors, such as ischemia and hypoxia, excessive light and hyperglycemia can cause the increase of reactive oxygen species and free radicals in the retina, thus inducing OS, damaging retina and affecting the normal visual function. Kelch-like ECH-associated protein 1(KEAP1)and nuclear factor erythroid 2 related factor 2(NRF2), which together constitute the main antioxidant stress signaling pathway in the body, play an antioxidant role by regulating retinal energy metabolism and cell proliferation, apoptosis and autophagy through various ways, so as to reduce retinal damage caused by OS. In this paper, the role and mechanism of the KEAP1-NRF2 signaling pathway regulation of OS in the retinal are briefly reviewed, aiming to provide ideas for subsequent research.

Humans ; Consensus ; Hypersensitivity ; Desensitization, Immunologic/adverse effects* ; Immunotherapy/adverse effects* ; Injections, Subcutaneous ; Allergens

AIM: To systematically evaluate the effects of low-energy red light on choroidal thickness(ChT), spherical equivalent(SE), and axial length(AL)in myopic children.METHODS: A total of 8 databases, including Pubmed, Embase, Cochrane Library, Web of Science, CNKI, WanFang Database, VIP Database and China Biomedical Literature Database, were electronically retrieved to collect the clinical randomized controlled trial(RCT)of low-energy red light in myopia, with red light intervention as an experimental group and glasses as a control group. The retrieval time limit is from the establishment of the database to January 2023. According to the recommendation of the Cochrane Handbook, literature quality and risk of bias were evaluated. Revman5.4 software was used for Meta-analysis.RESULTS: Totally 8 RCT about red-light treatment with myopia were included. The sample size for ChT analysis contained 407 eyes in the red-light group and 425 eyes in the control group; SE analysis included 490 eyes in the red-light group and 518 eyes in the control group; sample size for AL analysis were 518 eyes in the red-light group and 539 eyes in the control group. The differences in ChT, SE and AL between the groups were statistically significant(ChT: WMD=37.81, 95%CI: 21.55～54.07, P<0.001; SE: WMD=0.58, 95%CI: 0.46～0.71, P<0.001; AL: WMD=-0.31, 95%CI: -0.39～-0.24, P<0.001).CONCLUSION: Specific red light can promote the increase of ChT while inhibit SE and AL. However, its long-term efficacy and side effects remain unclear. The above conclusions need to be further clarified by more RCT with large samples.

OBJECTIVE: The study aimed to estimate the benchmark dose (BMD) of coke oven emissions (COEs) exposure based on mitochondrial damage with the mitochondrial DNA copy number (mtDNAcn) as a biomarker. METHODS: A total of 782 subjects were recruited, including 238 controls and 544 exposed workers. The mtDNAcn of peripheral leukocytes was detected through the real-time fluorescence-based quantitative polymerase chain reaction. Three BMD approaches were used to calculate the BMD of COEs exposure based on the mitochondrial damage and its 95% confidence lower limit (BMDL). RESULTS: The mtDNAcn of the exposure group was lower than that of the control group (0.60 ± 0.29 vs. 1.03 ± 0.31; P < 0.001). A dose-response relationship was shown between the mtDNAcn damage and COEs. Using the Benchmark Dose Software, the occupational exposure limits (OELs) for COEs exposure in males was 0.00190 mg/m ³. The OELs for COEs exposure using the BBMD were 0.00170 mg/m ³ for the total population, 0.00158 mg/m ³ for males, and 0.00174 mg/m ³ for females. In possible risk obtained from animal studies (PROAST), the OELs of the total population, males, and females were 0.00184, 0.00178, and 0.00192 mg/m ³, respectively. CONCLUSION Based on our conservative estimate, the BMDL of mitochondrial damage caused by COEs is 0.002 mg/m ³. This value will provide a benchmark for determining possible OELs.
Male ; Female ; Animals ; Coke ; Polycyclic Aromatic Hydrocarbons ; DNA Copy Number Variations ; Benchmarking ; Occupational Exposure/analysis* ; DNA, Mitochondrial/genetics* ; DNA Damage

Objective: To investigate the association between physical exercise and non-alcoholic fatty liver disease (NAFLD) in people infected with HBV. Methods: The information about the 3 813 participants infected with HBV, including the prevalence of NAFLD, prevalence of physical exercise and other covariates, were collected from the National Science and Technology Major Project of China during 2016-2020. The logistic regression model was used to evaluate the association between physical exercise and NAFLD in HBV infected patients, and subgroup analysis was performed to identify the effect modifiers. Results: A total of 2 259 HBV infected participants were included in the final analysis and 454 (20.10%) had NAFLD. After adjusting for covariates, we found that moderate physical exercise was a protective factor for NAFLD (OR=0.66, 95%CI: 0.46-0.94). Subgroup analysis suggested that the protective effect of moderate physical exercise on NAFLD might be stronger in women (OR=0.61, 95%CI: 0.36-1.01), those <45 years old (OR=0.24, 95%CI: 0.06-0.80), those who had low education level (OR=0.16, 95%CI: 0.04-0.49), those who had low annual income (OR=0.39, 95%CI: 0.16-0.89 for <30 000 yuan RMB; OR=0.64, 95%CI: 0.40-1.00 for 30 000-80 000 yuan RMB), those who had hypertension (OR=0.45, 95%CI: 0.21-0.88), those with BMI ≥24.0 kg/m² (OR=0.66, 95%CI: 0.43-1.01), those who had more daily fruit or vegetable intake (OR=0.61, 95%CI: 0.38-0.97), those who had more daily meat intake (OR=0.49, 95%CI: 0.23-0.97), and those who had no smoking history (OR=0.66, 95%CI: 0.45-0.95) or passive smoking exposure (OR=0.61, 95%CI: 0.37-0.97). Conclusions: Among HBV infected patients, moderate physical exercise was negatively associated with the prevalence of NAFLD. Women, young people, those who had low education level, those who had low annual income, those with hypertension, those with high BMI, those who had more daily fruit or vegetable and meat intakes, and those who had no smoking history or passive smoking exposure might be more sensitive to the protective effect.
Humans ; Female ; Adolescent ; Middle Aged ; Non-alcoholic Fatty Liver Disease/epidemiology* ; Hepatitis B virus ; Risk Factors ; Tobacco Smoke Pollution ; Exercise ; Hypertension

Aim To predict the potential mechanism of ophiopogonin D (OPD) against pulmonary fibrosis by network pharmacology, and further verify it by experiment in vivo. Methods This study found that ophiopogon was the most frequently used drug in the treatment of pulmonary fibrosis with deficiency of Qi and Yin through data mining. In order to explore its material basis, network pharmacology analysis was carried out. A model of pulmonary fibrosis was established by bleomycin, and different concentrations of ophiopogonin D were administered to verify the results of the pharmacological network. Results Firstly, through network pharmacology analysis, it was found that mitophagy might be the potential target for ophiopogon to exert anti-pulmonary fibrosis effect. Meanwhile, network topology analysis showed that OPD had the greatest relationship with mitophagy. Animal experiments showed that OPD could relieve pulmonary fibrosis and reduce collagen deposition in mice. At the same time, the detection of mitophagy related proteins showed that the compound could increase the expression of PINK1 and Parkin proteins, reduce the content of P62 protein in lung tissue, and reduce the intracellular ROS level. Conclusions OPD can improve mitochondrial function and play an anti-pulmonary fibrosis role by promoting PINKl/Parkin dependent mitophagy in lung tissue.

Aim To prepare prostate cancer exosomes containing melittin and observe their uptake by prostate cancer cells. Methods Cells treated with starvation for different time were screened for exosome extraction. Exosomes from PC-3 cells were extracted by ultracentrifugation, and the extracted particles were examined by transmission electron microscopy, nanoparticle tracking analyzer(NTA), and Western blot. Melittin exosome system was prepared by repeated freeze-thaw method, incubation at room temperature as well as electroporation, and the size of encapsulation efficiency was measured by centrifugation. A high-performance liquid chromatography(HPLC)method was applied to assay the content of melittin exosomes(exo-mel). Fluorescence inverted microscopy was employed to evaluate the uptake of melittin exosomes by PC-3 cells, DU145 cells as well as LNCaP cells. Results The results of starvation treatment showed that 24 h starvation treatment was the optimal time point. TEM results showed that the exosomes were round or oval in shape with a distinct membranous structure, and the diameter was around 100 nm. The reagent protein concentration for NTA analysis of exosomes was 0.222 g·L-1. The results of Western blot for the marker proteins of exosomes showed that Alix and CD63 were positively expressed, which indicated that the exosomes could be obtained by starvation culture of PC-3 cells and ultracentrifugation. The results of entrapment efficiency showed that the entrapment efficiency of electroporation method was 17.51% ± 2.39%, that of repeated freeze-thaw method was 11.46% ± 1.02%, and that of room temperature incubation method was 3.93% ± 2.44%. The encapsulation efficiency of electroporation was the highest with significant difference(P<0.05). The uptake assay showed that PC-3 cells could efficiently take up exo-mel in a time-dependent manner, and DU145 cells and LNCaP cells also could take up exo-mel over time. Conclusions Exosomes can be accessed by starvation treatment and high-speed centrifugation, and the prostate cancer melittin exosome system prepared by electroporation method could be taken up by prostate cancer cells.