Background Methicillin-resistant Staphylococcus aureus (MRSA) is an important pathogen for both human bloodstream infections and mastitis in cows. However, little attention has been paid to the cross-host transmission of MRSA from cows to high-risk groups in China. Objective To determine the MRSA colonization rates among dairy cows and dairy farm workers in Xinjiang, identify the antibiotic resistance profiles and molecular characteristics of the isolates, and provide scientific evidence for the formulation of targeted infection control strategies. Method A cross-sectional survey combined with laboratory pathogen analysis was conducted. From June to August 2024, large-scale dairy farms in Xinjiang region were selected as study sites. Nasal swabs (n=96) and skin swabs (n=39) were collected from workers, and bovine nasal swab samples (n=109) were collected simultaneously. All samples were subjected to MRSA isolation, cultivation, and identification, followed by antibiotic susceptibility testing to characterize resistance phenotypes. Staphylococcus aureus protein A (Spa) typing was performed to determine strain genotypes and elucidate MRSA colonization rates and molecular epidemiological patterns. Results A total of 35 MRSA strains was successfully isolated from 244 samples. The MRSA colonization rates among dairy farm workers and dairy cows were 20.83% (20/96) and 12.84% (14/109), respectively, with an overall isolation rate of 14.34% (35/244). Among the workers, the nasal colonization rate was 16.67% (16/96), and the skin colonization rate was 12.82% (5/39). One worker exhibited MRSA colonization at multiple body sites. All MRSA strains were resistant to cefoxitin (100%, 35/35). The resistance rates to erythromycin and clindamycin were 42.86% (15/35) and 34.29% (12/35), respectively. Thirteen strains showed a multidrug-resistant phenotype, whereas all strains were susceptible to vancomycin. The MRSA isolates exhibited high genetic diversity, with 13 Spa types identified, among which t441 was the most prevalent (8 strains). Both t441 and t034 types were detected in samples from both the dairy cows and their handlers. These two Spa types also carried and stably inherited specific resistance combinations, including erythromycin–clindamycin–cefoxitin and ciprofloxacin–erythromycin–clindamycin–gentamicin–cefoxitin–tetracycline, and a statistically significant association was also observed between the two resistance profiles and the bacterial types (P < 0.001). In addition, one novel Spa type strain was identified. Conclusion MRSA colonization rates among dairy cows and dairy farm workers in Xinjiang are relatively high, with evidence of multi-site colonization. The isolates exhibit high levels of multidrug resistance and genetic diversity, indicating a potential risk of cross-host transmission.

Background The high work intensity and possible subsequently increased susceptibility to occupational hazards of calcium carbide plants may lead to hypertension in workers, but there are few studies on the relationship between occupational hazard exposure and hypertension in workers involving the production process of calcium carbide. Objective To explore the influence of occupational hazards on the incidence of hypertension in calcium carbide plants. Methods Using historical cohort design, the employees of a calcium carbide factory in the western part of Inner Mongolia Autonomous Region were selected as research subjects. According to the pre-determined inclusion and exclusion criteria, the study population comprised an exposure group of 377 employees (including furnace workers, inspection workers, and maintenance workers) exposed to dust, noise & carbon monoxide, and a control group of 388 employees (including central control workers, electricians, and administrative personnel) without above-mentioned exposure. The total sample size was 765 participants. The follow-up period was from April 2011 to October 2022, and the study endpoint was defined as the conclusion of the follow-up period or diagnosed hypertension in annual occupational health examination. Information on general demographic characteristics, living habits, and work status was collected from all study subjects. Cox proportional hazards regression model was used to analyze the association between occupational hazard exposure and the risk of hypertension among the calcium carbide plant employees. Results The average age, mean systolic and diastolic blood pressure, proportion of males, smoking rate, and alcohol consumption rate in the exposure group were higher than those in the control group (P<0.05). Compared to baseline, both systolic and diastolic blood pressure levels increased in the exposure group and the control group at the end of the follow-up (P<0.05). At the end of the follow-up, the average differences between systolic/ diastolic blood pressure and baseline values in the exposure group were higher than those in the control group (P<0.05). During the follow-up period, a total of 223 cases of hypertension occurred, with a total follow-up of 2785 person-years, resulting in an incidence density of 8007.18 per 100000 person-years, an average age of onset of (35.90±8.22) years, and an average working age of onset of (2.69±1.97) years. The incidence density in the exposure group was 128.71% higher than that in the control group, and the risk of hypertension in the exposure group was 2.115 times that of the control group. The risk of hypertension was 2.199 times higher for men than for women, 1.344 times higher for those aged 30 and above than for those under 30, 1.546 times higher for smokers than for non-smokers, and 1.750 times higher for drinking workers than for non-drinking ones. The results of Cox proportional hazards regression modeling indicated that the hazard ratio (95%CI) of hypertension among the employees exposed to dust, noise, and carbon monoxide was 2.254 (1.703, 2.982), 1.594 (1.107, 2.295), and 1.567 (1.079, 2.274), respectively, when different covariates (gender, age, smoking, and alcohol consumption) were included. The results of Log-rank test showed that there were statistically significant differences in the distribution of disease-free survival between the exposure group and the control group (P<0.05). Conclusion Occupational exposures to dust, noise, and carbon monoxide may increase the risk of hypertension among calcium carbide plant workers.

OBJECTIVE To evaluate the impact of the specialized centralized procurement policy for insulin on daily cost and affordability of insulin， and provide data support for the enhancement of relevant policies. METHODS In this research， the insulin purchasing data were obtained from provincial centralized procurement platforms in provinces before and after the specialized centralized procurement of insulin （October-December 2021 and October-December 2022）， and the cost variations of insulin before and after the centralized procurement were analyzed by the defined daily dose cost （DDDc） of various types of insulins. The changes in the affordability of various types of insulins before and after the specialized centralized procurement were evaluated， using the percentage of annual expenditure on various types of insulins relative to annual per capita disposable income （i.e. the proportion of annual expenditure） as an indicator. RESULTS After the specialized centralized procurement， DDDc of various types of insulins decreased by 20.7%-71.8%， with an average reduction of 45.7%. Moreover， the reduction in DDDc for third-generation insulin exceeded that for second-generation insulin. The reduction in the proportion of annual expenditure on insulin ranged from 24.3% to 73.4%， with an average decrease of 48.5%. Premixed insulin analogs experienced the greatest reduction （73.4%）. Following the specialized centralized procurement， DDDc of insulin decreased in all provinces. Except for Guangxi （10.2%）， the average proportion of annual expenditure on insulin in the remaining provinces dropped to below 10%. CONCLUSIONS The specialized centralized procurement policy for insulin has significantly reduced insulin costs and improved affordability， thereby alleviating the economic pressure on patients with diabetes. There are notable cost disparities among provinces and among insulin categories， which require attention.

Bone repair remains an important target in tissue engineering, making the development of bioactive scaffolds for effective bone defect repair a critical objective. In this study, β-tricalcium phosphate (β-TCP) scaffolds incorporated with processed pyritum decoction (PPD) were fabricated using three-dimensional (3D) printing-assisted freeze-casting. The produced composite scaffolds were evaluated for their mechanical strength, physicochemical properties, biocompatibility, in vitro pro-angiogenic activity, and in vivo efficacy in repairing rabbit femoral defects. They not only demonstrated excellent physicochemical properties, enhanced mechanical strength, and good biosafety but also significantly promoted the proliferation, migration, and aggregation of pro-angiogenic human umbilical vein endothelial cells (HUVECs). In vivo studies revealed that all scaffold groups facilitated osteogenesis at the bone defect site, with the β-TCP scaffolds loaded with PPD markedly enhancing the expression of neurogenic locus Notch homolog protein 1 (Notch1), vascular endothelial growth factor (VEGF), bone morphogenetic protein-2 (BMP-2), and osteopontin (OPN). Overall, the scaffolds developed in this study exhibited strong angiogenic and osteogenic capabilities both in vitro and in vivo. The incorporation of PPD notably promoted the angiogenic-osteogenic coupling, thereby accelerating bone repair, which suggests that PPD is a promising material for bone repair and that the PPD/β-TCP scaffolds hold great potential as a bone graft alternative.
Calcium Phosphates/chemistry* ; Animals ; Bone Regeneration ; Rabbits ; Tissue Scaffolds ; Printing, Three-Dimensional ; Humans ; Human Umbilical Vein Endothelial Cells ; Neovascularization, Physiologic ; Osteogenesis ; Tissue Engineering/methods* ; Biomimetic Materials ; Cell Proliferation ; Angiogenesis

Objective:The objective of this study is to sequence whole genome of the rabies virus in the saliva specimen of a suspected rabies case with onset of rabies from a wildlife (hog badger) bite wound in Sichuan province, where the genetic variation characteristics of the virus was analyzed at the molecular level, so as to understand the prevalence and mutation of wildlife rabies virus in Sichuan province.Methods:Total viral RNA was extracted from the saliva specimen of the suspected rabies case. Then, rabies virus sequences were amplified using PCR with specific primers; the gene fragments obtained were sequenced, and the sequences obtained were spliced using biological software to obtain the whole genome sequence of the rabies virus strain. The genetic variation characteristics of the whole genome were analyzed.Results:The whole genome nucleotide sequence of a strain of rabies virus of hog badger origin (hereinafter referred to as SCR23-052) was obtained by sequencing, and NCBI online BLAST and comparison with several reference sequences showed that the composition and structure of the whole genome sequence of SCR23-052 conformed to the characteristics of the Lyssavirus under the Rhabdoviridae; the highest similarity in nucleotide and amino acid sequences in various gene regions was observed between SCR23-052 and the strains of Ningxia (J) and Chongqing (CQ92, 02050CHI). The sequence variability of SCR23-052 genome was significantly lower at the amino acid level than that at the nucleotide level, which indicated that most of the nucleotide variants in the protein-coding genes belonged to synonymous mutations. Phylogenetic analysis showed that SCR23-052 belonged to genotype V, which did not show any obvious mutation in the major antigenic site of the glycoprotein, underwent amino acid glycosylation at positions 56 and 338 by the online site prediction, and showed the least amino acid difference compared with the signal peptide sequence of the vaccine strain CTN181. The virus in this study has an A→T mutation at position 332 in the nucleoprotein major antigenic site with all reference vaccine strains, and an L→V mutation at position 379 in the B-cell epitope with the CTN181 vaccine strain. SCR23-052 was consistent with both genotype V reference strains at the nucleoprotein study site.Conclusions:The whole genome sequence of a wildlife strain of genotype V rabies virus of hog badger origin was obtained, which was different from that of the genotype I strain of rabies virus of dog origin that previously reported to be prevalent in Sichuan. The genome sequence of SCR23-052 differed from that of the reference vaccine strains to a varying degree, but the main virulence characteristics remained unaltered.

Objective:To investigate the differences between convolutional neural network (CNN) measurements and manual measurements of prosthetic angles and leg length discrepancies after total hip arthroplasty (THA).Methods:Data were collected from 100 patients who underwent unilateral THA at the Department of Orthopaedics, Fourth Medical Center of PLA General Hospital, between January and March 2024. Fifty patients received a ceramic-lined hip prosthesis, and 50 received a polyethylene-lined hip prosthesis. The cohort included 58 males and 42 females, with an average age of 51.6±12.71 years. The left hip was affected in 45 cases, and the right hip in 55 cases. The Attention UNet network was used to segment the prostheses and identify key points, from which the acetabular prosthesis abduction angle, anteversion angle, and leg length discrepancy were calculated. The CNN measurements of anteversion angle, abduction angle, and leg length discrepancy were compared with manual measurements. The consistency between the two methods was evaluated using the Bland-Altman method.Results:CNN measurements of the abduction angle, anteversion angle, and leg length discrepancy for the 100 prostheses were 40.55°±4.76°, 21.5°(19.0°, 27.0°), and 3.6(1.7, 6.8) mm, respectively, while manual measurements were 40.58°±4.93°, 21.0° (17.6°, 23.2°), and 3.0 (2.0, 7.0) mm, respectively. There was a statistically significant difference in anteversion angle between the two methods ( Z=-2.189, P=0.029), but no significant differences in abduction angle or leg length discrepancy ( P>0.05). For the 50 cases with polyethylene-lined prostheses, CNN-measured abduction and anteversion angles were 40.31°±4.70° and 20.0°(18.0°, 22.0°), respectively, while manual measurements were 40.49°±4.92° and 20.0°(17.0°, 21.5°); there were no statistically significant differences ( P>0.05). For the 50 ceramic-lined prostheses, CNN-measured abduction and anteversion angles were 40.79°±4.86° and 26.5°(20.8°, 33.0°), respectively, while manual measurements were 40.68°±4.98° and 20.0°(18.0°, 22.0°), with a significant difference in anteversion angle ( Z=-3.426, P=0.001) but not in abduction angle ( t=0.994, P=0.325). Bland-Altman analysis showed a difference of 0.031°±1.091° for the abduction angle of the 100 prostheses between the two methods, 0.178°±1.296° for the polyethylene-lined prostheses, and -0.116°±0.825° for the ceramic-lined prostheses. For the anteversion angle, the difference was -3.280°±6.920° for the 100 prostheses, -0.002°±1.471° for the polyethylene-lined prostheses, and -6.560°±8.523° for the ceramic-lined prostheses. The difference in leg length discrepancy was -0.101±0.947 mm. Conclusion:CNN measurements of acetabular prosthesis abduction angle, polyethylene-lined prosthesis anteversion angle, and leg length discrepancy after THA showed no significant differences from manual measurements. However, CNN and manual measurements of ceramic-lined prosthesis anteversion angle differed. Bland-Altman analysis indicates that the two methods have good consistency when measuring the abduction angle, the lower limb discrepancy, and the anteversion angle of the polyethylene liner prosthesis.

BACKGROUND:Bone tissue defects are one of the most common diseases in orthopedics,and the current treatments for this disease are inadequate.The development of tissue engineering brings new hope for bone defect repair:by regulating the release of bioactive substances and the process of vascularization and neurogenesis at the defect site,it can effectively improve the microenvironment of bone tissue and promote osseointegration,which is the most promising research idea for large-size bone defect repair. OBJECTIVE:To explore the research progress of regulating bone microenvironment changes in bone defect repair in recent years from the effects of bioactive substances,vascularization and neurotization on three aspects of bone microenvironment changes,and to provide new ideas and strategies for the treatment of large-size bone defects. METHODS:The search terms"bone tissue engineering,angiogenesis,neurotization,cytokines,bone morphogenetic protein,vascular endothelial growth factor,neuropeptides,bone microenvironment"in Chinese and English were used to search for articles on the influence of changes in the bone microenvironment and their application in bone tissue engineering published from January 1,2001 to December 31,2022 on CNKI,WanFang,Web of Science,Science Direct,and PubMed.Finally,109 articles were included for review. RESULTS AND CONCLUSION:(1)The bone microenvironment is essential for the induction of bone tissue stem cell growth and differentiation,and mainly consists of the extracellular matrix of the bone tissue seeds and the biochemical factors required for intercellular interactions,the local blood circulation network and the surrounding nerve tissue.(2)Bone defect repair is a continuous process divided into multiple phases that overlap and are mediated by multiple cytokines,and the same cytokine can have mutually synergistic or antagonistic effects in one or more healing phases.(3)Neovascular regeneration is key to initiating bone repair,as neovascularisation not only provides essential nutrients,osteoblasts and growth factors for bone repair,but is also a gateway for repair cells to enter the injury zone.(4)In addition to regulating the type,dose and timeliness of vascular-inducing factor release to achieve blood transport reconstruction.The study of differential release delivery systems of multiple factors and the application of gene transfer technology will be the future research direction to solve large bone defects.(5)Neuropeptides can bind to relevant receptors and act on specific signaling pathways to guide vascular growth and influence bone healing,bone regeneration and the balance between osteogenesis and osteolysis through a variety of pathways.(6)In the establishment of neuralized tissue-engineered bone,the role of changes in the bone tissue microenvironment and neuromodulation is bidirectional.Cytokines in the bone matrix can participate in neuronal signaling pathways through the blood-nerve barrier.Neuropeptides secreted by glial cells act on the bone microenvironment,affecting bone healing,bone regeneration and the balance between osteogenesis and osteolysis.(7)There are still many questions regarding the regulation of the bone microenvironment by bioactive substances and the processes of vascularization and neurogenesis,such as the rapid diffusion and degradation of cytokines in the body and their loss of activity,the temporal and spatial distribution of angiogenesis-related growth factors,and the establishment of neurogenesis through the body's feedback regulatory mechanism,which need to be improved by subsequent studies.

OBJECTIVE To investigate the correlation between serum levels of asprosin and adropin in elderly patients with obstructive sleep apnea syndrome(OSAS)and the disease severity.METHODS From August 2021 to August 2023,131 elderly OSAS patients admitted to Ezhou Central Hospital were collected as observation subjects(OSAS group),according to the sleep apnea hypopnea index(AHI),there were 40 cases in the mild group,52 cases in the moderate group,and 39 cases in the severe group,meantime,124 healthy individuals who came to health clinic of our hospital for physical examination were collected as the control group.Enzyme-linked immunosorbent assay(ELISA)was applied to determine the levels of asprosin and apropin in the serum of OSAS patients;Pearson method was applied to analyze the correlation between serum asprosin,apropin levels and AHI,ODI,and LSaO2.ROC curve was used to analyze the diagnostic value of serum asprosin and adropin levels in OSAS and the severity of OSAS patients.RESULTS Compared with the control group,the asprosin level in the OSAS group was obviously increased,while the adropin level was obviously reduced(P<0.05).There was no statistically obvious difference in gender,age,coronary heart disease,hypertension,TC,TG,HDL-C,LDL-C among the mild,moderate,and severe groups(P>0.05);compared with the mild group,the levels of BMI,AHI,ODI,and asprosin in the moderate and severe groups were obviously increased,while the levels of LSaO2 and apropin were obviously reduced(P<0.05);compared with the moderate group,the levels of BMI,AHI,ODI,and asprosin in the severe group were obviously increased,while the levels of LSaO2 and apropin were obviously reduced(P<0.05).The serum asprosin level in OSAS patients was positively correlated with AHI and ODI,and negatively correlated with LSaO2(P<0.05);the level of apropin was negatively correlated with AHI and ODI,and positively correlated with LSaO2(P<0.05).The AUC of serum asprosin,apropin levels,and their combination in diagnosing moderate and severe OSAS was 0.832,0.882,and 0.942,respectively,the combined diagnostic value of the two was superior to that of single diagnosis(Z=3.435,2.560,P=0.001,0.011).Serum asprosin,adropin levels and AUC of combined diagnosis of OSAS were 0.818,0.804 and 0.893,respectively.The value of combined diagnosis was better than that of single diagnosis(Z=3.886,4.126,P=0.000,0.000).CONCLUSION The serum level of asprosin is increased and the level of adropin is decreased in patients with OSAS,which is closely related to the severity of the disease,and may be used in the clinical diagnosis of OSAS and the evaluation of the severity of OSAS.

Objective:To establish a cell line stably expressing the transient receptor potential melastatin 2(TRPM2)channel for screening TRPM2 inhibitors based on PiggyBac transposition system.Methods:A plasmid PiggyBac-human TRPM2(pPB-hTRPM2)eukaryotic expression vector was constructed using PiggyBac transposition system.The plasmid and a helper plasmid were co-transfected into HEK293T cells to express TRPM2,which was identified by fluorescence and patch-clamp assays.The high throughput screening performance was assessed with the Z'factor.Calcium imaging and patch clamp techniques were employed to assess the initial activity of eleven compound molecules,confirming the inhibitory effects of the primary molecules on TRPM2.The protective effect of the screened compounds on damaged cells was validated using the oxygen-glucose deprivation/reperfusion(OGD/R)injury model and CCK-8 kit.The level of cellular reactive oxygen species(ROS)was detected by flow cytometry.The neuroprotective effects of the compounds were evaluated using a transient middle cerebral artery occlusion(tMCAO)mouse model.Results:The HEK293T cells transfected with pPB-hTRPM2-EGFP showed high TRPM2 expression.Puromycin-resistant cells,selected through screening,exhibited robust fluorescence.Whole-cell patch results revealed that induced cells displayed classical TRPM2 current characteristics comparable to the control group,showing no significant differences(P＞0.05).With a Z'factor of 0.5416 in calcium imaging,the model demonstrated suitability for high-throughput screening of TRPM2 inhibitors.Calcium imaging and electrophysiological experiments indicated that compound 6 significantly inhibited the TRPM2 channel.Further experiments showed that 1.0 μmol/L of compound 6 enhanced cell viability(P＜0.05)and reduced the level of ROS(P＜0.05)of SH-SY5Y under OGD/R injury.0.3 and 1.0 mg/kg of compound 6 reduced the cerebral infarction volume in tMCAO mice(both P＜0.05).Conclusions:A stable TRPM2 gene expressing cell line has been successfully established using PiggyBac gene editing in this study.TRPM2 channel inhibitors were screened through calcium imaging and patch clamp techniques,and an inhibitor compound 6 was identified.This compound can alleviate cell damage after OGD/R by reducing cellular ROS levels and has a protective effect against cerebral ischemia-reperfusion injury in mice.

AIM:To investigate the effect of curcumin on the viability and apoptosis of human papillomavirus(HPV)-positive cervical cancer cells via down-regulating the expression of adenovirus E1A-associated 300 kD protein(P300).METHODS:HeLa cells in logarithmic phase were divided into four experimental groups(20,40,60 and 80 μmol/L Cur treatment groups),and the untreated group was used as control.Cell apoptosis was detected by flow cytome-try,and the expression of E6 gene was detected by RT-qPCR and Western blot.Silencing plasmid(shE6)and negative control shRNA(shNC)were constructed and transfected into HeLa cells,the cells were randomly divided into four groups including shNC,shNC+Cur(40 μmol/L),shE6 and shE6+Cur(40 μmol/L).The cell viability,apoptosis and expression of apoptosis-related proteins were detected by CCK-8,flow cytometry and Western blot,respectively.Then silencing plas-mid(siP300)and negative control siRNA(siNC)were constructed and transfected into HeLa cells.The mRNA and pro-tein expression of P300 was detected by RT-qPCR and Western blot.The protein expression of E6 and P300 and histone was detected by Western blot after treated with 20 and 40 μmol/L curcumin.RESULTS:Compared with the untreated group,the HeLa cells treated with different concentrations of curcumin could significantly inhibit the viability and increase the early apoptosis rate(P<0.05).The results showed that the mRNA and protein expression of E6 were down-regulated in HeLa cells after treated with different concentration curcumin,while the early apoptosis and the expression of pro-apop-tosis-related proteins in the shE6 group were lower than those in the non-knockdown group after treated with 40 μmol/L cur-cumin(P<0.05).And the expression of E6 protein was decreased after knockdown of P300,while the expression of P300 and histone was down-regulated after treatment with 20 and 40 μmol/L curcumin.CONCLUSION:Curcumin can inhibit the viability and promote the apoptosis of HPV18 positive cervical cancer cells,and the mechanism may be that it can in-hibit E6 acetylation by down-regulating P300 expression.