Background Environmental metal exposure is closely associated with the onset and progression of mild cognitive impairment (MCI) in the elderly. Effectively identifying hazardous metal exposure and assessing their interaction effects have significant public health implications. Objective To explore the relationship between urinary single metal and metal mixture exposure and MCI in elderly compound residents. Methods This study included 391 elderly individuals aged 60 and above from residential compounds in Zhongshan City, Guangdong Province. Concentrations of iron (Fe), copper (Cu), selenium (Se), arsenic (As), cadmium (Cd), manganese (Mn), chromium (Cr), nickel (Ni), vanadium (V), cobalt (Co), antimony (Sb), thallium (Tl), zinc (Zn), calcium (Ca), and magnesium (Mg) in urine were measured using inductively coupled plasma mass spectrometry. Cognitive function in the elderly was assessed using the Chinese version of the Mini-Mental State Examination (MMSE). Logistic regression was used to explore the relationship between single metal exposure level and MCI. LASSO regression and multi-metal logistic regression models were used to identify key metal ions associated with MCI. Bayesian kernel machine regression (BKMR) was employed to analyze the relationship between key metal ion mixtures and MCI, as well as the interactions between metals. Age, gender, education level, occupation, and body mass index were adjusted as covariates. Results A total of 78 among the 391 elderly individuals surveyed (19.94%) were diagnosed with MCI (MCI group), and the other 313 individuals were controls. The levels of Se, Cd, Mn, and As in the urine of the MCI group were significantly higher than those in the control group (P < 0.05). In the single-metal model, after adjusting for covariates and using the first quartile (Q₁) of each metal concentration as the reference, the OR for MCI in the elderly in the Q₄ group of Se was 2.190 (95%CI: 1.017, 4.716); for Cd, the OR was 2.345 (95%CI: 1.041, 5.283) in the Q₃ group and 2.371 (95%CI: 1.043, 5.393) in the Q₄ group; for Mn, the OR was 2.355 (95%CI: 1.038, 5.344) in the Q₂ group; for As, the OR was 3.377 (95%CI: 1.442, 7.908) in the Q₃ group and 2.886 (95%CI: 1.227, 6.788) in the Q₄ group; for Sb, the OR was 2.779 (95%CI: 1.234, 6.257) in the Q₂ group. When urinary metal concentrations were ln-transformed and included as continuous variables in the single-metal model, Cd concentration was positively correlated with MCI (OR=1.377; 95%CI: 1.008, 1.882; P=0.044). Cd, Se, Mg, Ca, Mn, As, Cr, Co, Tl, and Sb were selected by the LASSO regression model and included in the multi-metal model. In the multi-metal model, compared with Q₁, the OR for MCI in the elderly was 0.395 (95%CI: 0.164, 0.953) in the Q₂ group of Co and 0.390(95%CI: 0.167，0.911) in the Q₃ group of Co; for Mn, the OR in the Q₂ group was 2.636 (95%CI: 1.053, 6.596); for Sb, the OR in the Q₂ group was 2.640 (95%CI: 1.047, 6.658). As continuous variables, Mg (OR=0.472; 95%CI: 0.248, 0.899; P=0.022) and Co (OR=0.857; 95%CI: 0.737, 0.996; P=0.044) concentrations were negatively correlated with MCI. The BKMR mixture analysis suggested that Mg and Co exhibited a synergistic negative correlation with MCI, while Mn and Sb exhibited a synergistic positive correlation with MCI. Mg and Co attenuated the positive correlation of Mn and Sb with MCI, whereas Mn weakened the protective effects of Mg and Co. Conclusion Elevated levels of Se, Cd, As, Mn, and Sb in urine may increase the risk of MCI in the elderly, while Mg and Co have protective effects. Potential synergistic or antagonistic interactions may be found among Mn, Sb, Mg, and Co, which should not be overlooked in terms of their impact on the cognitive function of the elderly.

BackgroundDementia seriously affects the quality of life and lifespan of elderly people， with Alzheimer's disease （AD） being the most common type of dementia. Previous studies have suggested that gout may reduce the risk of developing AD， but the causal relationship between the two still requires further research. ObjectiveTo investigate the potential causal relationship between gout and AD through a two-sample Mendelian randomization （MR） analysis， so as to provide references for the prevention and treatment of AD. MethodsData from Genome-Wide Association Studies （GWAS） extracted in 2024 were analyzed， using pooled data on gout （6 810 cases in the case group and 477 788 cases in the control group） published by UK Biobank in 2021 as the exposure variable， and data on AD （3 899 cases in the case group and 214 893 cases in the control group） published by FinnGen in the same year as the outcome variable. The inverse-variance weighted， MR-Egger regression， weighted median estimation， simple model and weighted model were used to analyze the potential causal relationship between gout and AD. Pleiotropic effects were assessed using MR-Egger regression. Heterogeneity assessment was conducted using Cochran's Q test. The leave-one-out analysis was carried out for sensitivity analysis. And a funnel plot was drawn to detect potential publication bias. ResultsThe inverse-variance weighted analysis demonstrated a negative causal relationship between gout and AD （OR=0.004， 95% CI： 0~0.700， P<0.05）. The plot resembled a symmetrical inversed funnel， indicating the absence of publication bias. No heterogeneity was detected by Cochran's Q test. The MR-Egger regression indicated no significant horizontal pleiotropy. Concerning the reverse directions， no significant associations between AD and gout were noted. ConclusionThere is a negative causal relationship between gout and AD， with gout potentially reducing the risk of developing AD. ［Funded by The Third Batch of Social Welfare and Basic Research Projects （Medical and Health） of Zhongshan City in 2022 （number， 2022B3017）］

Objective Vasculogenic mimicry(VM)is a novel vasculogenic process integral to glioma stem cells(GSCs)in glioblastoma(GBM).However,the relationship between VM and ataxia-telangiectasia mutated(ATM)serine/threonine kinase activation,which confers chemoradiotherapy resistance,remains unclear. Methods We investigated VM formation and phosphorylated ATM(pATM)levels by CD31/GFAP-periodic acid-Schiff dual staining and immunohistochemical staining in 145 GBM specimens.Glioma stem-like cells(GSLCs)derived from the formatted spheres of U87 and U251 cell lines and their pATM level and VM formation ability were examined using western blot and three-dimensional culture.For the examination of the function of pATM in VM formation by GSLCs,ATM knockdown by shRNAs and deactivated via ATM phosphorylation inhibitor KU55933 were studied. Results VM and high pATM expression occurred in 38.5%and 41.8%of tumors,respectively,and were significantly associated with reduced progression-free and overall survival.Patients with VM-positive GBMs exhibited higher pATM levels(rs=0.425,P=0.01).The multivariate analysis established VM as an independent negative prognostic factor(P=0.002).Furthermore,GSLCs expressed high levels of pATM and formed vascular-like networks in vitro.ATM inactivation or knockdown hindered VM-like network formation concomitant with the downregulation of pVEGFR-2,VE-cadherin,and laminin B2. Conclusion VM may predict a poor GBM prognosis and is associated with pATM expression.We propose that pATM promotes VM through extracellular matrix modulation and VE-Cadherin/pVEGFR-2 activation,thereby highlighting ATM activation as a potential target for enhancing anti-angiogenesis therapies for GBM.

Objective:To establish a disease risk prediction model for the newborn screening system of inherited metabolic diseases by artificial intelligence technology.Methods:This was a retrospectively study. Newborn screening data ( n=5 907 547) from February 2010 to May 2019 from 31 hospitals in China and verified data ( n=3 028) from 34 hospitals of the same period were collected to establish the artificial intelligence model for the prediction of inherited metabolic diseases in neonates. The validity of the artificial intelligence disease risk prediction model was verified by 360 814 newborns ' screening data from January 2018 to September 2018 through a single-blind experiment. The effectiveness of the artificial intelligence disease risk prediction model was verified by comparing the detection rate of clinically confirmed cases, the positive rate of initial screening and the positive predictive value between the clinicians and the artificial intelligence prediction model of inherited metabolic diseases. Results:A total of 3 665 697 newborns ' screening data were collected including 3 019 cases ' positive data to establish the 16 artificial intelligence models for 32 inherited metabolic diseases. The single-blind experiment ( n=360 814) showed that 45 clinically diagnosed infants were detected by both artificial intelligence model and clinicians. A total of 2 684 cases were positive in tandem mass spectrometry screening and 1 694 cases were with high risk in artificial intelligence prediction model of inherited metabolic diseases, with the positive rates of tandem 0.74% (2 684/360 814)and 0.46% (1 694/360 814), respectively. Compared to clinicians, the positive rate of newborns was reduced by 36.89% (990/2 684) after the application of the artificial intelligence model, and the positive predictive values of clinicians and artificial intelligence prediction model of inherited metabolic diseases were 1.68% (45/2 684) and 2.66% (45/1 694) respectively. Conclusion:An accurate, fast, and the lower false positive rate auxiliary diagnosis system for neonatal inherited metabolic diseases by artificial intelligence technology has been established, which may have an important clinical value.

Current evidence shows that subcutaneous injection of methotrexate results in more rapid and better absorption, higher blood drug levels and less variable exposure than oral dosing. Among patients with rheumatoid arthritis or psoriasis, subcutaneous methotrexate is more effective than oral methotrexate, and has a lower risk of gastrointestinal complaints. For patients in whom oral methotrexate is proved to be inadequate or intolerant, subcutaneous methotrexate also shows good efficacy. Subcutaneous methotrexate can serve as a treatment option before the use of biological agents, and in this way, large amounts of money can be saved. Subcutaneous methotrexate has good application prospects for clinical practice.

Objective:To investigate the expression of mucin-4 (MUC4) in meningiomas.Methods:Totally 258 cases of meningiomas and 165 cases of other brain tumors were collected from the First Affiliated Hospital of China University of Science and Technology (Anhui Provincial Hospital) from 2011 to 2017. MUC4, EMA, PR, SSTR-2 protein expression was detected by immunohistochemistry, and their expression in meningiomas and other tumor tissue was compared.Results:The 258 patients with meningioma included 85 males and 173 females, with a mean age of 69 years. Among the meningiomas, 192, 54 and 12 were WHO grades Ⅰ, Ⅱ and Ⅲ respectively. The overall expression rate of MUC4 in meningiomas was 67.8% (175/258), including 46/46 (100.0%) in meningothelial meningiomas, 3/3 in secretory meningiomas, 44/45 (97.8%) in angiomatous meningiomas, 37/41 (90.2%) in atypical meningiomas, 3/4 in metaplastic meningiomas, 2/3 in microcystic meningiomas, 7/11 in psammomatous meningiomas, 7/11 in chordoid meningiomas, 14/28 (50.0%) in transitional meningiomas, 1/2 in clear cell meningiomas, 1/2 in papillary meningiomas, 4/9 in anaplastic meningiomas, 7/52 (13.5%) in fibrous meningiomas, and 0/1 in rhabdoid meningiomas. In addition, MUC4 was expressed in 44 EMA negative meningiomas and in four SSTR-2 negative meningiomas. PR, EMA, SSTR-2 were expressed in 149 cases (57.7%), 173 cases (67.1%), 235 cases (91.1%) of meningiomas, respectively. MUC4 was not expressed in other tumors in the central nervous system, including schwannomas, neurofibromas, solitary fibrous tumors/hemangiopericytoma (SFT/HPC), hemangioblastoma, gliomas and ependymomas.Conclusion:MUC4 is widely expressed in meningiomas and has great value in distinguishing meningiomas from other non-meningeal epithelial tumorsof the central nervous system.

BACKGROUND: Cervical intervertebral foramen stenosis induced by cervical spondylosis of nerve root type usual y requires surgical treatment. The ways mainly include anterior cervical discectomy and fusion and cervical posterior intervertebral foramen decompression. Which is the best way is stil inconclusive. With innovation, anterior cervical discectomy and fusion for cervical spondylosis of nerve root type has become the mainstream in the current treatment. OBJECTIVE: To study the relationship between curative effects and intervertebral foramen-associated parameter changes in patients with cervical spondylosis of nerve root type after anterior cervical discectomy and fusion. METHODS: From March 2011 to April 2013, 132 patients with cervical spondylosis of nerve root type were treated with anterior cervical discectomy and fusion in the Changzheng Hospital Affiliated to the Second Military Medical University. Neck pain and arm pain visual analogue score, neck disability index score and imaging changes were evaluated before and after treatment. RESULTS AND CONCLUSION: 132 patients were fol owed up for 25(4-28) months. Significant differences in neck pain visual analogue scale, anterior intervertebral disc height, posterior intervertebral disc height, intervertebral foramen height, anterior and posterior diameters of the intervertebral foramen, the area of the intervertebral foramen, and the Cobb angle of the fused segment were detected in al patients before and after treatment (P < 0.05). Posterior intervertebral disc height was positively correlated with intervertebral foramen area (r=0.427, P=0.000). The increased Cobb angle of the fused segment was negatively associated with the size of intervertebral foramen (r=-0.273, P=0.003). Intervertebral foramen area was negatively associated with arm pain visual analogue score (r=-0.502, P=0.000). These results indicated that anterior cervical discectomy and fusion with an interbody fusion cage can obviously enlarge intervertebral foramen in patients with cervical spondylosis of nerve root type, and obtain good curative effect. The size of the intervertebral foramen is negatively related to the axial pain. The reconstruction of the intervertebral disc height is necessary to expand the intervertebral foramen. However, the increase of the curvature fusion segments is not helping to expand the intervertebral foramen.

BACKGROUND:Intervertebral disc degeneration is one of the ancient and common clinical diseases. Its complex pathogenesis affected by various factors, such as environment and genes, is stil in debate. Because of the technical limitations, there is stil no deep understanding on the molecular mechanism of intervertebral disc degeneration. However, its molecular mechanism in recent years has made considerable development. OBJECTIVE: To summarize and discuss the molecular mechanism of intervertebral disc degeneration, thereby providing the basis for the effective treatment. METHODS: CNKI and Medline databases were retrieved by the first author using computer to search relevant articles published from 2005 to 2015. The key words were “intervertebral disc degeneration, molecular mechanism, environmental factors, genes, matrix, degradation enzyme,inflammatory factor, biological environment, treatment” in Chinese and English, respectively. Mechanisms of intervertebral disc degeneration, involving genes, cel senescence and apoptosis, degradation enzyme and substrate, inflammatory cytokines, were summarized to explore the pathogenesis and possible effective treatment of intervertebral disc degeneration. RESULTS AND CONCLUSION:Totaly 153 articles were initialy retrieved and finaly 52 articles were included in result analysis according to inclusive and exclusive criteria. Unique structure and biochemical properties of the intervertebral disc are easy to cause intervertebral disc degeneration. Traditionaly, environmental factors, such as occupation and smoking, are considered as the main factors inducing intervertebral disc degeneration; however, more and more studies have shown that genes have the most important influence on intervertebral disc degeneration. Declined extracelular matrix, increased degradation enzymes, and overexpression of inflammatory factors can al destroy the entire structure of intervertebral disc, and accelerate the process of intervertebral disc degeneration. Effective treatment for intervertebral disc degeneration can be formulated depending on the deep understanding on its molecular mechanisms. Although there is a further understanding on the molecular medium of intervertebral disc degeneration, the complex biochemical environment within the intervertebral disc is stil a great chalenge to the treatment of intervertebral disc degeneration.

OBJECTIVETo elucidate the role of transforming growth factor-beta1(TGF-β1) in epithelial-mesenchymal transition of mesothelial cells and peritoneal metastasis of gastric cancer.

METHODSHMrSV5 cells, a human peritoneal mesothelial cell line, were incubated with TGF-β1, and their morphological changes were observed by phase contrast microscopy. Expressions of α-smooth muscle actin (α-SMA), vimentin, cytokeratin, E-cadherin, phosphorylated-Smad2 and Smad2 were examined by Western blotting. After fibroblastic-like mesothelial cells were co-incubate with HSC-39 cells(gastric cancer cell line), the adhesion and invasion potential of HSC-39 were evaluated by adhesion and invasion assay in vitro.

RESULTSFew mesothelial cells converted to spindle fibroblast-like morphology for 24 h, and remarkable phenotypic changes were observed at 72 h of TGF-β1 activation. TGF-β1 could induce α-SMA and vimentin expression, and down-regulate cytokeratin and E-cadherin expression in mesothelial cells (P<0.05). TGF-β1 induced phosphorylation of Smad2 within 15 min of stimulation, reached a maximum at 30 min after treatment and remained high level during the experiment without affecting total Smad2 expression(P>0.05). The percentage of HSC-39 gastric cancer cells adhered were significantly increased as compared to the control. When the mesothelial cells were treated by TGF-β1 for 72 h, the increased adhesion percentage was(146±17)%(P<0.05). After fibroblastic-like mesothelial cells co-incubated with HSC-39 cells for 48 h, more cancer cells [(61.1±11.4) cells/view field] invaded the coated membrane as compared to the control group [(31.9±8.1) cells/view field] (P<0.05).

CONCLUSIONTGF-β1 can induce the transition of mesothelial cells into myofibroblasts and Smad2 signal pathway may play a role in this transition, which is associated with increased adhesion and invasiveness of gastric cancer cells, and provides favorable environment for the dissemination of gastric cancer.

Cadherins ; Cell Line, Tumor ; Epithelial Cells ; Epithelial-Mesenchymal Transition ; Epithelium ; Fibroblasts ; Humans ; Peritoneal Neoplasms ; Signal Transduction ; Smad2 Protein ; Stomach Neoplasms ; Transforming Growth Factor beta1 ; Vimentin

Objective To construct a recombinant Bacillus Calmette-Guerin ( BCG ) vaccine strain, rBCG::Rv3478-pMV261, expressing the Rv3478 protein of Mycobacterium tuberculosis and to inves-tigate its immunogenicity.Methods The gene fragments encoding Rv3478 antigen were amplified by PCR and then respectively cloned into pMV261 and pET-28a vectors to construct the recombinant expression plas-mids (Rv3478-pMV261 and Rv3478-pET-28a).The Rv3478-pMV261 plasmids were transformed into the BCG cells to construct the rBCG vaccine strains, while the Rv3478-pET-28a plasmids were transformed into Escherichia coli BL21 strains for the expression of Rv3478 protein.Polyclonal antibodies were induced in mice upon the immunization with Rv3478 protein.The rBCG vaccine strains overexpressing Rv3478 protein were screened out with Western blot assay.The C57BL/6 mice were divided into four groups including the PBS treated group, BCG treated group, rBCG::pMV261 ( R0) treated group and rBCG::Rv3478-pMV261 ( R3) treated group.All mice were sacrificed in 4 or 12 weeks after immunization.Enzyme-linked immunos-pot assay ( ELISPOT) , ELISA and flow cytometry analysis were performed to evaluate the induced humoral and cell-mediated immune responses in mice.Results The Rv3478 protein was successfully expressed and could induce polyclonal antibodies in mice.High levels of IFN-γand TNF-αwere detected in mice treated with R3, indicating that the immunization with R3 enhanced the cellular immunity.Moreover, the ratios of CD4+to CD8+T cells and the percentages of CD44+CD62L+T cells were increased in mice upon the immuni-zation with R3.Conclusion The recombinant BCG vaccine strain overexpressing Rv3478 protein could in-duce stronger cell-mediated immune responses in mice.It might be have a great significance as a new tuber-culosis( TB) vaccine strain against TB infection in the future.