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.

Objective:To carry out an overview of systematic reviews on interventions to prevent the recurrence of diabetic foot ulcer (DFU) patients.Methods:Systematic reviews or Meta-analysis on interventions to prevent DFU recurrence were electronically retrieved from databases such as China National Knowledge Infrastructure, WanFang Data, VIP, China Biology Medicine disc, Joanna Briggs Institute (JBI) Evidence-Based Healthcare Center Database, Cochrane Library, PubMed, Embase, CINAHL and Web of Science. The search period was from database establishment to April 20, 2023. Two researchers independently searched and screened literature, and extracted data, and used AMSTAR 2 software and the quality evaluation criteria for systematic review of JBI Evidence-Based Healthcare Center to evaluate the methodological quality of the included literature. GRADE evaluation system was used for quality assessment of outcome indicators (DFU recurrence rate, effectiveness of measures to prevent DFU recurrence) .Results:A total of 24 systematic reviews were included. Studies showed that monitoring and intervention of foot skin temperature, therapeutic shoes or insoles, comprehensive intervention measures, and specific surgical methods could reduce the recurrence rate of DFU, while foot self-care, foot exercise, health education, and psychological intervention had no statistical effect on preventing DFU recurrence. The methodological quality of systematic reviews was generally low. The reports of four articles were relatively complete, 18 articles had certain defects, and two articles had serious defects. The evaluation of evidence quality showed that there were three pieces of moderate-quality evidence, seven pieces of low-quality evidence, and 30 pieces of extremely low-quality evidence.Conclusions:Existing evidence suggests that foot skin temperature monitoring and intervention, therapeutic shoes or insoles, comprehensive intervention measures, and specific surgical methods (such as Achilles tendon lengthening, metatarsophalangeal joint replacement, metatarsal head resection and so on) have a positive impact on preventing DFU recurrence. Rigorous and high standard research is still needed to verify the controversial issues, so as to provide reliable evidence for future clinical practice and studies.

Objective:To investigate the role of group 3 innate lymphoid cells (ILC3) in skin wound healing, and to explore the underlying mechanisms.Methods:Twenty-four 5-week-old male C57BL/6 mice were randomly and equally allocated into 3 groups: the skin wound + ILC3 inhibitor group (referred to as ILC3 inhibitor group), the skin wound group, and the control group, with 8 mice in each group. Four days before the establishment of the wound model, mice in the ILC3 inhibitor group were intraperitoneally injected with 1 μg of ILC3 inhibitor every 2 days for a total of 2 doses, mice in the skin wound group were injected with an equal volume of physiological saline solution, and mice in the control group were fed normally. To establish a mouse skin wound model, a full-thickness circular incision with a diameter of 0.6 cm was made around the midpoint of the dorsal midline using a biopsy punch after the intraperitoneal injection of anesthetics, which was histologically confirmed to be a full-thickness injury. The size of the wounds was observed and recorded, photographs of the wounds were taken on days 0, 1, 3, 5, 7, and 9 after wounding, and corresponding wound healing rates were calculated. On day 9 after wounding, tissue samples were collected from the wound edges, and subjected to flow cytometry analysis to quantify ILC3 infiltrating around the skin wound, and hematoxylin and eosin (HE) staining was performed to assess the healing status of the skin wounds. Real-time quantitative polymerase chain reaction (qRT-PCR) was conducted to determine the mRNA expression of vitamin D receptor (VDR), Notch1, tumor necrosis factor-alpha (TNF-α), interleukin (IL) -17A, IL-17F, and IL-22 in the wound-edge tissues, and Western blot analysis to determine their protein expression. Statistical analysis was carried out by using one-way analysis of variance and t test. Results:On day 9 after wounding, the skin wound group showed an increased number of ILC3 in the wound-edge tissues (5.31% ± 1.47% vs. 3.10% ± 0.54%, P < 0.01), increased mRNA and protein expression of TNF-α, IL-22, IL-17A, and IL-17F (all P < 0.05), but decreased mRNA and protein expression of VDR (both P < 0.05) compared with the control group; the protein expression of Notch1 was significantly higher in the skin wound group than in the control group ( P < 0.05), but there was no significant difference in its mRNA expression between the two groups ( P > 0.05). On days 1, 3 and 5, the wound healing rates were significantly higher in the ILC3 inhibitor group (45.17% ± 9.90%, 61.58% ± 11.61%, 75.61% ± 9.12%, respectively) than in the skin wound group (25.87% ± 10.96%, 47.78% ± 13.81%, 64.55% ± 10.29%, respectively, all P < 0.05). On day 9, the ILC3 inhibitor group showed a decreased number of ILC3 around the wound (2.69% ± 0.95%, P < 0.01), decreased mRNA and protein expression of TNF-α, IL-22, IL-17A, and IL-17F in the wound-edge tissues (all P < 0.05), but increased mRNA and protein expression of Notch1 and VDR in the wound-edge tissues (all P < 0.05) compared with the skin wound group. On day 9 after wounding, histopathological examination with HE staining revealed continuous and intact epithelial structure, as well as dense and neatly arranged collagen fibers in the ILC3 inhibitor group, and the structures of hair follicles, blood vessels, and sebaceous glands were similar to those in the control group. Conclusions:Skin ILC3 infiltrated local wounds and were involved in the skin wound healing process through inflammatory factors such as TNF-α, IL-17A, IL-17F, and IL-22. Downregulating the number of ILC3 may promote skin wound healing by activating VDR and Notch1, as well as inhibiting the TNF-α signaling pathway and the expression of downstream inflammatory factors.

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.

Objective To observe the value of 18F-FDG PET/CT semi-quantitative parameters for predicting spread through air spaces(STAS)of clinical stage Ⅰa—Ⅲa lung adenocarcinoma.Methods Data of 85 patients with clinical stage Ⅰa—Ⅲ a lung adenocarcinoma who underwent preoperative 18F-FDG PET/CT were retrospectively analyzed.The patients were divided into positive group(n=23)or negative group(n=62)according to whether pathology showed STAS or not.Clinical and PET/CT data were compared between groups,and logistic analysis was performed to explore the efficacy of each parameter for predicting STAS.Results Significant differences of gender,carcinoma embryonic antigen,clinical stage,pathological grade,micropapillary growth and proportion were found between groups(all P＜0.05).The maximum,the mean,the peak standard uptake value(SUVmax,SUVmean,SUVpeak),as well as the maximum,the mean and the peak standard uptake value normalized by lean body mass(SULmax,SULmean,SULpeak),also the total lesion glycolysis(TLG)in positive group were all significantly higher than those in negative group(all P＜0.05).Patients'gender,proportion of micropapillary growth,SUVmax and SULmax were all independent risk factors of STAS of clinical stage Ⅰa—Ⅲa lung adenocarcinoma.The area under the curve(AUC)of the above parameters for predicting STAS was 0.666,0.912,0.839 and 0.842,respectively,and of the combination was 0.957.Conclusion 18 F-FDG PET/CT semi-quantitative parameters SUVmax and SULmax were helpful for predicting STAS of clinical stage Ⅰa—Ⅲ a lung adenocarcinoma,and further combination of gender and proportion of micropapillary growth could improve diagnostic efficacy.

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.

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: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 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.