ObjectiveSpinal cord injury (SCI) directly impairs the regulatory function of the autonomic nervous system, induces intestinal dysfunction, and significantly reduces patients’ quality of life. Preclinical studies have shown that electroacupuncture (EA) therapy can regulate the brain-gut axis and is used to treat central nervous system diseases such as major depressive disorder, Alzheimer’s disease and Parkinson’s disease. Recent research has established that fecal microbiota transplantation (FMT) from EA-treated SCI rats restored intestinal motility and colonic morphology. However, it remains unclear whether the regulation of gut microbiota by EA therapy directly contributes to neural repair after SCI. This study aims to explore whether gut microbiota mediates the neuroprotective effect of EA in the treatment of SCI and its possible mechanism. MethodsThe study employed RNA transcriptome analysis of spinal cord tissue to characterize gene expression profiles and to identify key signaling pathways following EA treatment for SCI. Hematoxylin-Eosin (HE) staining and Nissl staining were used to observe the morphological changes in spinal cord tissue. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the effects of EA on the expression of proteins related to nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) -dependent pyroptosis. Using 16S rDNA sequencing, the study observed alterations in gut microbiota diversity and community composition in SCI rats. Prior to establishing SCI models, rats were pretreated with an antibiotic cocktail to induce gut dysbiosis, and the effects on intestinal function and spinal cord neural repair were evaluated. FMT was performed to investigate the regulatory effects of post-EA FMT on motor function, general status, liver and spleen indices, and NLRP3-mediated pyroptosis in SCI rats. ResultsEA improved motor function and reduced regulated neuronal cell death in SCI rats. Transcriptomic analysis demonstrated the activation of immune- and inflammation-related pathways post-SCI, including NOD-like receptors, nuclear factor-kappa B(NF-κB), and Toll-like receptor (TLR) pathways. EA primarily influenced intestinal inflammation and autoimmune functions. 16S rDNA sequencing illustrated that EA did not alter the diversity of gut microbiota. However, EA altered the gut microbiota composition in SCI rats, increasing Lactobacillus and Akkermansia genera while rebalancing the Firmicutes/Bacteroidetes ratio. Furthermore, depletion of gut microbiota by antibiotics disrupted the intestinal barrier, reduced the expression of intestinal barrier proteins Zonula Occludens-1 (ZO-1) and Occludin, elevated serum lipopolysaccharide-binding protein (LBP) levels, exacerbated spinal cord tissue damage, and hindered motor function recovery in SCI rats. FMT from donors treated with EA reduced LBP levels in the intestine, blood, and spinal cord of rats, inhibited the TLR4 myeloid differentiation primary response protein 88 (MyD88)-NF‑κB pathway and NLRP3-dependent pyroptosis, and improved motor function. On the other hand, FMT treatment resulted in decreased body weight and food intake, whereas FMT using EA-treated donors effectively alleviated these alterations. ConclusionEA effectively alleviated neuroinflammatory responses in rats with SCI, primarily through regulating the gut microbiota and suppressing the NLRP3-dependent pyroptosis signaling pathway.

The inflammatory microenvironment is closely associated with the initiation and progression of osteoarthritis （OA）， specifically manifesting as macrophage activation， dysregulation of inflammatory cytokines， and redox imbalance. Following an overview of the pathological characteristics of the OA inflammatory microenvironment， this paper reviews the research progress on the mechanism of traditional Chinese medicine （TCM） intervening in OA by modulating the inflammatory microenvironment. It has been found that TCM monomers/active ingredients （such as total alkaloids from Strychnos nux-vomica ， quercetin， triptolide， etc.）， herb pairs （e.g. Angelica pubescens - Gentiana macrophylla ， Carthami Flos-Lycopodii Herba）， and TCM formulas （such as Zhuanggu jianxi formula， Duhuo jisheng decoction and Rongjin niantong formula， etc.） can inhibit macrophage activation， reduce the release of proinflammatory cytokines and the generation of reactive oxygen species by inhibiting multiple signaling pathways， including nuclear factor-κB， Wnt/ β -catenin， and mitogen-activated protein kinase， thereby alleviating the articular inflammatory microenvironment， restoring local joint homeostasis， and slowing the progression of OA.

Objective This study reports on the obligatory parasitism of water mites Arrenurus sp. on Anopheles messeae at the Manzhouli Port, Inner Mongolia, China. Methods Duing July 2024, a survey on the mosquito diversity was conducted at the Manzhouli Port. Captured mosquitoes and their ectoparasites were identified to species level. Results A total of 1840 adult mosquitoes were collected, representing species from three genera: Culex(Cx. modestus, Cx. pipiens pallens), Aedes(Ae. dorsalis, Ae. flavidorsalis, Ae. flavescens), and Anopheles (An. messeae). Among all the mosqutioes specimens,3 out of 150 captured An. messeae were found to carry ectoparasitic mites, with number of 2,4,27 mites separately. Morphological and molecular identification reached the same result as water mites(Hydrachnidiae, Hydracrina). COI gene sequence showed 94% similarity with the closest species Arrenurus truncatellus. Conlusions Literature review suggests water mites are host-specific parasitism of mosquito species and herein with the first record of Arrenurus sp. parasiting on An. Messeae in the most high-latitude region globally.

ObjectiveSpinal cord injury (SCI) directly impairs the regulatory function of the autonomic nervous system, induces intestinal dysfunction, and significantly reduces patients’ quality of life. Preclinical studies have shown that electroacupuncture (EA) therapy can regulate the brain-gut axis and is used to treat central nervous system diseases such as major depressive disorder, Alzheimer’s disease and Parkinson’s disease. Recent research has established that fecal microbiota transplantation (FMT) from EA-treated SCI rats restored intestinal motility and colonic morphology. However, it remains unclear whether the regulation of gut microbiota by EA therapy directly contributes to neural repair after SCI. This study aims to explore whether gut microbiota mediates the neuroprotective effect of EA in the treatment of SCI and its possible mechanism. MethodsThe study employed RNA transcriptome analysis of spinal cord tissue to characterize gene expression profiles and to identify key signaling pathways following EA treatment for SCI. Hematoxylin-Eosin (HE) staining and Nissl staining were used to observe the morphological changes in spinal cord tissue. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the effects of EA on the expression of proteins related to nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) -dependent pyroptosis. Using 16S rDNA sequencing, the study observed alterations in gut microbiota diversity and community composition in SCI rats. Prior to establishing SCI models, rats were pretreated with an antibiotic cocktail to induce gut dysbiosis, and the effects on intestinal function and spinal cord neural repair were evaluated. FMT was performed to investigate the regulatory effects of post-EA FMT on motor function, general status, liver and spleen indices, and NLRP3-mediated pyroptosis in SCI rats. ResultsEA improved motor function and reduced regulated neuronal cell death in SCI rats. Transcriptomic analysis demonstrated the activation of immune- and inflammation-related pathways post-SCI, including NOD-like receptors, nuclear factor-kappa B(NF-κB), and Toll-like receptor (TLR) pathways. EA primarily influenced intestinal inflammation and autoimmune functions. 16S rDNA sequencing illustrated that EA did not alter the diversity of gut microbiota. However, EA altered the gut microbiota composition in SCI rats, increasing Lactobacillus and Akkermansia genera while rebalancing the Firmicutes/Bacteroidetes ratio. Furthermore, depletion of gut microbiota by antibiotics disrupted the intestinal barrier, reduced the expression of intestinal barrier proteins Zonula Occludens-1 (ZO-1) and Occludin, elevated serum lipopolysaccharide-binding protein (LBP) levels, exacerbated spinal cord tissue damage, and hindered motor function recovery in SCI rats. FMT from donors treated with EA reduced LBP levels in the intestine, blood, and spinal cord of rats, inhibited the TLR4 myeloid differentiation primary response protein 88 (MyD88)-NF‑κB pathway and NLRP3-dependent pyroptosis, and improved motor function. On the other hand, FMT treatment resulted in decreased body weight and food intake, whereas FMT using EA-treated donors effectively alleviated these alterations. ConclusionEA effectively alleviated neuroinflammatory responses in rats with SCI, primarily through regulating the gut microbiota and suppressing the NLRP3-dependent pyroptosis signaling pathway.

Objective To evaluate the impact of incorporating two doses of varicella vaccine into the immunization program in Shanghai on the incidence of varicella, age-specific incidence, and varicella outbreaks in kindergartens and schools in Jinshan District, and to assess its prevention and control effects. Methods Descriptive epidemiological methods were used to analyze and compare the population distribution characteristics and the occurrence of varicella outbreaks before (2013-2017) and after (2018-2024) the implementation of the strategy. Interrupted time series (ITS) design and its segmented regression model were used to analyze the changes in the incidence of varicella in the two stages before and after the implementation of the vaccination of two doses of varicella vaccine, and to evaluate the short-term and long-term preventive and control effects of the strategy. Results After the incorporation of two doses of varicella vaccine into the immunization program, the average annual incidence of varicella in Jinshan District decreased from 82.19/100 000 (2013-2017) to 53.51/100 000 (2018-2024). The interrupted time series analysis showed that after the implementation of the strategy, the slope change in the incidence of varicella was -10.20/100 000 (t=-2.38, P=0.04), and the overall trend was downward (slope β₁+β₃= -8.34/100 000). Except for the 30-49 years old group, the incidence rates in all other age groups decreased to varying degrees. Among them, the 5-9 years old group had a better vaccine efficacy (VE) of 52.23% (95% CI: 49.07-60.65), followed by the 10-14 years old group. The number of potential varicella outbreaks and cluster outbreaks in nurseries, kindergartens, and schools across the district dropped significantly. Conclusion Incorporating two doses of varicella vaccine into the immunization program can effectively reduce the incidence of varicella and the number of varicella outbreaks in kindergartens and schools, which is of great significance for the prevention and control of varicella.

Objective: To explore the impacts of coal mine dust on lung function in rats, so as to provide the basis for the early prevention and treatment of coal worker's pneumoconiosis. Methods: Seventy-two SPF-grade 8-week-old male Sprague-Dawley rats were randomly divided into the coal dust group, the coal-silica dust group, the silica dust group and the control group. The rats in the first three groups of rats were administered 1 mL corresponding dust suspension into the lungs using non-exposure tracheal instillation, while the rats in the control group were administered 1 mL normal saline. Respiratory rate (f), forced vital capacity (FVC), peak expiratory flow (PEF) and dynamic pulmonary compliance (Cdyn) were measured at 1, 3 and 6 months after dust exposure. Lung tissues were collected to measure reactive oxygen species (ROS) and adenosine triphosphate (ATP) levels using corresponding ELISA kits and ATP assay kits, respectively. The relative mRNA expressions of peroxisome proliferators-activated receptor gamma coactivator 1-alpha (PGC-1α) and mitochondrial transcription factor A (TFAM) were detected using real-time fluorescent quantitative polymerase chain reaction assay. The relative protein expressions of PGC-1α and TFAM were detected using Western blotting. Results: There was no interaction between dust type and exposure duration on f (P>0.05), but there were interactions on FVC, PEF and Cdyn (all P<0.05). Compared with the control group at 6 months after dust exposure, the f of the rats in the silica dust group were increased, while the FVC and PEF of the rats in the coal-silica dust and silica dust groups were decreased, and Cdyn of the rats in the coal dust, coal-silica dust and silica dust groups were decreased (all P<0.05). There were interactions between dust type and exposure duration on ROS and ATP levels, the relative mRNA and protein expressions of PGC-1α and TFAM (all P<0.05). Compared with the control group at 3 and 6 months after dust exposure, the ROS levels in the rats in the coal dust, coal-silica dust and silica dust groups were increased, while the ATP levels, the relative mRNA and protein expressions of PGC-1α and TFAM were decreased (all P<0.05). Conclusion The lung function impairment in rats caused by different types of coal mine dust is related to PGC-1α-mediated mitochondrial biogenesis dysfunction, which leads to increased ROS levels, decreased ATP and TFAM levels.

Objective To explore the role of miR-1260b targeting activating transcription factor 6β(ATF6β)in osteogenic differentia-tion of human periodontal ligament stem cells(hPDLSCs).Methods The periodontal ligament tissue was scraped from the third molar extracted from the patient with periodontitis,and hPDLSCs were isolated and cultured.The proportions of positive cells of CD34,CD45,CD29,CD90 and CD105 were detected by flow cytometry,and the expression of vimentin and pan-cytokeratin were detected by immunofluo-rescence staining.The cellular luciferase activities of ATF6β-WT and ATF6β-MUT in the miR-1260b mimic+WT/MUT group(transfected with miR-1260b mimic and ATF6β-WT/MUT)and the mimic-NC group(transfected with miR-NC and ATF6β-WT/MUT)were analyzed by the dual luciferase reporter gene assay.In addition,the cells were divided into the miR-1260b mimic group(transfected with miR-1260b mimic alone),the ATF6β-OE group(transfected with the ATF6β overexpression vector alone),the combined group(transfected with miR-1260b mimic and ATF6β overexpression vector simultaneously),and the NC group(transfected with empty vector).Alizarin red S staining and alkaline phosphatase staining were performed on hPDLSCs in the NC group,the miR-1260b mimic group,the ATF6β-OE group and the combined group.Meanwhile,the mRNA expression levels of miR-1260b,ATF6β,Runt-related transcription factor 2(Runx2),osteocalcin(OCN),and osteopontin(OPN)in each group of cells were detected by qRT-PCR.The protein expressions of glucose-regulated protein 78(GRP78),C/EBP homologous protein(CHOP),and protein kinase R-like endoplasmic reticulum kinase(PERK)in each group of cells were detected by Western blot.Results The flow cytometry detection showed that CD29,CD90 and CD105 on the surface of the isolated and cultured cells were positive,while CD34 and CD45 were negative.Immunofluorescence staining showed that vimentin in the cells was positive and pan-cytokeratin was negative,which was in line with the characteristics of hPDLSCs.Luciferase assay showed that miR-1260b mimic could significantly reduce the luciferase activity of ATF6β-WT(P<0.05),but had no significant effect on the luciferase activity of ATF6β-MUT(P>0.05).After alizarin red S staining,the staining of hPDLSCs gradually deepened after 3 days,5 days and 7 days of culture,suggesting an enhanced osteogenic differentiation ability.With the enhancement of osteogenic differentiation ability of hPDLSCs,the expression of miR-1260b in the cells showed a gradually increasing trend,while the mRNA and protein expression levels of ATF6β showed a gradually decreasing trend(P<0.05).The results of alizarin red S staining and alkaline phosphatase staining showed that the proportions of positive area in the miR-1260b mimic group were higher than those in the NC group(P<0.05),while the proportions of positive area in the ATF6β-OE group and the combined group were lower than those in the NC group(P<0.05).The mRNA and protein levels of ATF6β in the miR-1260b mimic group were lower than those in the NC group(P<0.05),while the mRNA and protein levels of ATF6β in the ATF6β-OE group and the combined group were higher than those in the NC group(P<0.05).The mRNA levels of Runx2,OCN and OPN in the miR-1260b mimic group were significantly higher than those in the NC group(P<0.05).The mRNA levels of Runx2,OCN and OPN in the ATF6β-OE group and the combined group were significantly lower than those in the NC group(P<0.05).The protein expression levels of GRP78,CHOP and PERK in the miR-1260b mimic group were significantly lower than those in the NC group(P<0.05).The protein expression levels of GRP78,CHOP and PERK in the ATF6β-OE group and the combined group were significantly higher than those in the NC group(P<0.05).Conclusion miR-1260b can inhibit endoplasmic reticulum stress level and promote osteogenic differentiation of hPDLSCs by targeting ATF6β.

Objective To explore the clinical characteristics as well as molecular epidemiological features of resis-tance genes and virulence genes of carbapenem-resistant Klebsiella pneumoniae(CRKP)infection in a region,and provide scientific basis for the prevention,treatment,and epidemiological study of CRKP.Methods 60 non-repeti-tive CRKP strains isolated clinically from Puyang Oilfield General Hospital from November 2023 to September 2024 were analyzed retrospectively.Antimicrobial susceptibility testing was performed using VITEK 2 Compact automa-tic microbial analyzer,K-B disk diffusion method,and micro-broth dilution method.Mucus phenotype of bacterial strains was identified by string test.Carbapenemase was detected by carbapenemase inhibitor enhancement assay.Molecular features,such as multi-locus sequence typing(MLST),capsule serotypes,resistance genes,virulence genes,plasmid replication types of strains,as well as the genetic and evolutionary relationships of strains were de-termined by whole genome sequencing and bioinformatics analysis.Results CRKP strains were mainly isolated from elderly male hospitalized patients.Specimens were mostly from sputum(71.67％),mainly distributed in depart-ment of respiratory medicine(30.00％).All strains were highly resistant to multiple commonly used antimicrobial agents,only with high susceptibility rates to cefotaxime/avibactam,tigecycline,and polymyxin B(＞60.00％).Two CRKP strains were positive for string test.95.00％of the strains produced class A serine carbapenemase.All strains carried fluoroquinolone,phosphomycin,β-lactam,and aminoglycoside resistance genes;enterobactin,Esche-richia coli common pilus(ECP),and outer membrane protein-related virulence genes;as well as plasmids from the IncF plasmid family.Carbapenemase gene was mainly blaKPC-2(95.00％),and the major capsule serotype was KL19(43.33％).In MLST,ST11(51.67％)was the dominant clone group,and ST11-KL62(n=12)was the dominant subtype.Conclusion CRKP in this hospital is highly resistant to multiple commonly used antimicrobial agents,and its mechanism of resistance to carbapenems is mainly related to the presence of blaKPC-2 resistance gene.All strains have coexisting multiple resistance genes and virulence genes,and show a phenomenon of multi-clone transmission.ST11 is the dominant clone group,and ST11-KL62 is the main prevalent subclone type.

Objective:To investigate the temporal and spatial distribution characteristics of fluoride in drinking water in rural areas of Shaanxi Province.Methods:From April 2019 to September 2023, rural drinking water monitoring sites were set up in all 108 agriculture-related counties (cities and districts) in Shaanxi Province. The ex-factory water and tap water were collected annually in dry season and wet season for water fluoride determination and water fluoride exceeding standard evaluation, and the exceedance rates of water fluoride were compared according to different years, regions, water period types, water sample types, water source types, and water treatment methods.Results:A total of 31 221 water samples were collected in Shaanxi Province from 2019 to 2023, and the median water fluoride was 0.370 mg/L, ranging from 0.001 to 6.220 mg/L. The exceedance rates of water fluoride in each year were 5.93% (370/6 236), 3.21% (200/6 232), 3.05% (191/6 260), 3.64% (228/6 256), and 2.36% (147/6 237), respectively. The differences between different years were statistically significant (χ 2 = 132.31, P < 0.001). The exceedance rates of water fluoride in Guanzhong, Northern Shaanxi, and Southern Shaanxi were 5.10% (563/11 044), 5.93% (376/6 339), and 1.42% (197/13 838), respectively, with statistically significant differences between different regions (χ 2 = 355.76, P < 0.001). The exceedance rates of water fluoride during the dry season and wet season were 3.68% (574/15 615) and 3.60% (562/15 606), respectively, with no statistically significant difference between the two water periods (χ 2 = 0.12, P = 0.724). The fluoride exceedance rates of ex-factory water and tap water were 3.15% (442/14 024) and 4.04% (694/17 197), respectively. The fluoride exceedance rate of tap water was higher than that of ex-factory water, and the difference was statistically significant (χ 2 = 17.21, P < 0.001). The fluoride exceedance rates of surface water and groundwater were 0.37% (39/10 588) and 5.32% (1 097/20 633), respectively. The fluoride exceedance rate of groundwater was higher than that of surface water, and the difference was statistically significant (χ 2 = 488.68, P < 0.001). The fluoride exceedance rates of conventional treatment, sedimentation and filtration, disinfection only, and untreated water samples were 1.21% (111/9 174), 2.82% (130/4 604), 3.56% (309/8 686), and 6.69% (586/8 757), respectively. There was a statistically significant difference between different water treatment methods (χ 2 = 396.04, P < 0.001). Conclusions:The fluoride exceedance rate of drinking water in rural areas of Shaanxi Province has decreased from 2019 to 2023, and the exceedance water samples are mainly concentrated in Northern Shaanxi and Guanzhong regions. There are differences in the fluoride exceedance rate of water samples in different years, regions, water sample types, water source types, and water treatment methods.