1.Prevalence and associated factors of work-related musculoskeletal disorders among workers in a manganese enterprise
Tianzi SHAN ; Junxiang MA ; Tian CHEN ; Kang NONG ; Yucheng SUN ; Xueting WANG ; Gaoman ZHANG ; Teng MA ; Zhuoran XIA ; Fengtao CUI ; Li CHEN ; Yanyan ZHENG ; Piye NIU
Journal of Environmental and Occupational Medicine 2026;43(3):333-340
Background Work-related musculoskeletal disorders (WMSDs) are a major occupational health concern, particularly among workers exposed to adverse ergonomic conditions. Manganese production involves heavy physical demands, yet research on WMSDs among manganese workers remains limited. Objective To investigate the prevalence and influencing factors of WMSDs among manganese workers in a manganese enterprise in Guangxi. Methods A cross-sectional survey was conducted from May to June 2024 on workers at a manganese factory in Guangxi. The Chinese Musculoskeletal Disorders Questionnaire was used to collect information on demographic characteristics, distribution of musculoskeletal symptoms, and work-related exposures. χ2 test was applied to compare differences in positive WMSDs rates across groups, and logistic regression analysis was performed to identify associated factors. Results A total of 1476 workers were enrolled in the study after pre-determined inclusion and exclusion criteria. The overall prevalence of WMSDs was 34.15%. The most commonly affected body regions were the lower back (17.28%), neck (16.67%), and shoulders (13.82%). The results of logistic regression analysis indicated that female, older age, and education level of college or above were associated with a higher risk of WMSDs (P<0.05). Awkward working postures were significantly associated with WMSDs in corresponding body regions; in particular, awkward postures of the neck, upper limbs, trunk, and lower limbs were related to an increased risk of WMSDs in multiple body sites (P<0.05). In addition, poor lighting conditions, high workplace temperature, frequent or sustained arm support during work, and high job demands were associated with an increased risk of overall or site-specific WMSDs (P<0.05). Conclusion The high prevalence of WMSDs among manganese workers is closely associated with demographic characteristics, working postures, and work environment and organizational factors. Targeted ergonomic interventions focusing on high-risk body regions and key ergonomic exposures are warranted to reduce the risk of WMSDs among manganese workers.
2.Effect of virtual reality biofeedback training combined with oral positioning therapy on dysphagia after oral cancer surgery
Mingxia XU ; Hui ZHU ; Piaopiao CHEN ; Kexin MENG ; Jie CHEN ; Jing CHEN ; Huifang SUN ; Yanyan SUN
Chinese Journal of Rehabilitation Theory and Practice 2026;32(4):445-452
ObjectiveTo explore the application of virtual reality biofeedback training combined with oral localization therapy in dysphagia after oral cancer surgery. MethodsFrom May, 2023 to July, 2024, 86 patients with dysphagia after oral cancer surgery in Zhejiang Provincial People's Hospital were randomly divided into control group (n = 43) and experimental group (n = 43). The control group received conventional swallowing function training, while the experimental group added virtual reality biofeedback training combined with oral positioning therapy, for four weeks. The Standardized Swallowing Function Assessment Scale (SSA), Functional Oral Intake Scale (FOIS) and M.D.Anderson Dysphagia Inventory (MDADI) were used for evaluation before intervention, and two weeks, four weeks and eight weeks after intervention. ResultsFor scores of SSA , the main effects of group (F = 150.190, P < 0.001, η2p = 0.641) and time (F = 230.870, P < 0.001, η2p = 0.733), as well as the interaction effect (F = 16.910, P < 0.001, η2p = 0.168) were all significant. For scores of FOIS, the main effects of group (F = 59.601, P < 0.001, η2p = 0.415) and time (F = 89.464, P < 0.001, η2p = 0.516), as well as the interaction effect (F = 7.990, P < 0.001, η2p = 0.087) were all significant. For scores of MDADI, the main effects of group (F = 33.133, P < 0.001, η2p = 0.283) and time (F = 49.650, P < 0.001, η2p = 0.371), as well as the interaction effect (F = 3.224, P = 0.023, η2p = 0.037) were all significant. ConclusionVirtual reality biofeedback training combined with oral localization therapy could improve the swallowing function, oral feeding ability and overall quality of life of patients with dysphagia after oral cancer surgery.
3.Mechanism of Tibetan Medicine Sanwei Doukoutang to Improve Cognitive Dysfunction in 5×FAD Mice Based on Wnt/β-catenin Signaling Pathway
Shuran LI ; Yaxin WANG ; Jing SUN ; Lei BAO ; Zihan GENG ; Dan XIE ; Ronghua ZHAO ; Yanyan BAO ; Qiyue SUN ; Jingsheng ZHANG ; Xinwei WANG ; Xinying LI ; Xihe CUI ; Xiaowei YANG ; LIUXIAN ; Mengyao CUI ; Qingshan LIU ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):54-60
ObjectiveTo explore the effects of the Tibetan medicine Sanwei Doukoutang (SWDK) on cognitive dysfunction in mice suffering from Alzheimer's disease (AD) and its related mechanism. MethodsFifty SPF 5 × FAD mice were randomly divided into model group, total ginsenoside group(0.04 g·kg-1), high-, medium-, and low-dose groups of SWDK (32.60, 16.30, 8.15 g·kg-1), with 10 mice in each group, and ten wild-type mice of the same age were used as the normal group, male and female in 1∶1. Gavage administration was performed once daily for 8 weeks. The Morris water maze test and contextual fear memory experiment were used to observe learning and memory function. Hematoxylin-eosin (HE) staining was utilized to observe the changes in the pathomorphology of brain tissue in mice. The levels of synaptophysin (SYP) and postsynaptic dense substance 95 (PSD95) in mice serum were detected by enzyme-linked immunosorbent assay (ELISA). The positive expression of brain-derived neurotrophic factor(BDNF) in the dentate gyrus (DG) region of mouse brain tissue was observed by immunohistochemistry (IHC). The protein levels of BDNF, Wnt family member 3A(Wnt3a), and β-catenin were detected in the hippocampus of mice by Western blot. ResultsCompared with the normal group of mice, the model group of mice had significantly more complex swimming routes and lower swimming speed (P<0.01), significantly lower percentage of time spent in the target quadrant (P<0.01), and a significantly lower percentage of freezing time (P<0.05). The number of neurons in the hippocampal region of mice was obviously reduced and unevenly arranged. The levels of SYP and PSD95(P<0.01) in the serum of mice were reduced, and the positive expression of BDNF in the DG region of the brain tissue of mice was reduced. The levels of hippocampal BDNF, Wnt3a, and β-catenin proteins in the hippocampus of mice were obviously reduced (P<0.05, P<0.01). Compared with the model group, the mice in the SWDK group and the total ginsenoside group had significantly shorter swimming routes, the high- and medium- dose SWDK groups significantly higher swimming speeds (P<0.01), significantly higher percentage of time spent in the target quadrant (P<0.01), obviously higher percentage of Freezing time (P<0.05), and obviously more neurons in the hippocampal region of the mice with tighter arrangement. The mice had elevated levels of serum SYP (P<0.05, P<0.01), PSD95 (P<0.01), increased BDNF-positive cells in the DG region of brain tissue, and obviously elevated levels of BDNF, Wnt3a, and β-catenin proteins in the hippocampus of mice (P<0.05, P<0.01). ConclusionSWDK can significantly improve the cognitive dysfunction of AD mice, and its mechanism may be related to regulating the Wnt/β-catenin signaling pathway, which promotes BDNF expression and thereby enhances synaptic plasticity, allowing neuronal signaling to be restored.
4.Mechanism of Tibetan Medicine Sanwei Doukoutang to Improve Cognitive Dysfunction in 5×FAD Mice Based on Wnt/β-catenin Signaling Pathway
Shuran LI ; Yaxin WANG ; Jing SUN ; Lei BAO ; Zihan GENG ; Dan XIE ; Ronghua ZHAO ; Yanyan BAO ; Qiyue SUN ; Jingsheng ZHANG ; Xinwei WANG ; Xinying LI ; Xihe CUI ; Xiaowei YANG ; LIUXIAN ; Mengyao CUI ; Qingshan LIU ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):54-60
ObjectiveTo explore the effects of the Tibetan medicine Sanwei Doukoutang (SWDK) on cognitive dysfunction in mice suffering from Alzheimer's disease (AD) and its related mechanism. MethodsFifty SPF 5 × FAD mice were randomly divided into model group, total ginsenoside group(0.04 g·kg-1), high-, medium-, and low-dose groups of SWDK (32.60, 16.30, 8.15 g·kg-1), with 10 mice in each group, and ten wild-type mice of the same age were used as the normal group, male and female in 1∶1. Gavage administration was performed once daily for 8 weeks. The Morris water maze test and contextual fear memory experiment were used to observe learning and memory function. Hematoxylin-eosin (HE) staining was utilized to observe the changes in the pathomorphology of brain tissue in mice. The levels of synaptophysin (SYP) and postsynaptic dense substance 95 (PSD95) in mice serum were detected by enzyme-linked immunosorbent assay (ELISA). The positive expression of brain-derived neurotrophic factor(BDNF) in the dentate gyrus (DG) region of mouse brain tissue was observed by immunohistochemistry (IHC). The protein levels of BDNF, Wnt family member 3A(Wnt3a), and β-catenin were detected in the hippocampus of mice by Western blot. ResultsCompared with the normal group of mice, the model group of mice had significantly more complex swimming routes and lower swimming speed (P<0.01), significantly lower percentage of time spent in the target quadrant (P<0.01), and a significantly lower percentage of freezing time (P<0.05). The number of neurons in the hippocampal region of mice was obviously reduced and unevenly arranged. The levels of SYP and PSD95(P<0.01) in the serum of mice were reduced, and the positive expression of BDNF in the DG region of the brain tissue of mice was reduced. The levels of hippocampal BDNF, Wnt3a, and β-catenin proteins in the hippocampus of mice were obviously reduced (P<0.05, P<0.01). Compared with the model group, the mice in the SWDK group and the total ginsenoside group had significantly shorter swimming routes, the high- and medium- dose SWDK groups significantly higher swimming speeds (P<0.01), significantly higher percentage of time spent in the target quadrant (P<0.01), obviously higher percentage of Freezing time (P<0.05), and obviously more neurons in the hippocampal region of the mice with tighter arrangement. The mice had elevated levels of serum SYP (P<0.05, P<0.01), PSD95 (P<0.01), increased BDNF-positive cells in the DG region of brain tissue, and obviously elevated levels of BDNF, Wnt3a, and β-catenin proteins in the hippocampus of mice (P<0.05, P<0.01). ConclusionSWDK can significantly improve the cognitive dysfunction of AD mice, and its mechanism may be related to regulating the Wnt/β-catenin signaling pathway, which promotes BDNF expression and thereby enhances synaptic plasticity, allowing neuronal signaling to be restored.
5.Huaihuasan Alleviates Inflammatory Responses in Rat Model of Ulcerative Colitis Due to Dampness-heat in Large Intestine by Regulating TLR4/NF-κB Signaling Pathway
Yanyan WANG ; Weiyi SUN ; Zuowu XI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(24):1-10
ObjectiveThis study aims to systematically evaluate the therapeutic effects of Huaihuasan on ulcerative colitis (UC) in the rats with the syndrome of dampness-heat in the large intestine and explore the potential mechanisms of action. MethodsA rat model of UC due to dampness-heat in the large intestine was successfully established by combining traditional Chinese medicine etiology with trinitrobenzenesulfonic acid (TNBS)/ethanol solution. Rats were randomly allocated into a normal group, a model group, a high-dose (10.58 g·kg-1·d-1) Huaihuasan group, a medium-dose (5.29 g·kg-1·d-1) Huaihuasan group, a low-dose (2.65 g·kg-1·d-1) Huaihuasan group, and a sulfasalazine (0.35 g·kg-1·d-1) group. The body weight changes and the diarrhea and bloody stool scores of the rats in each group were recorded. In addition, pathological changes of the colon tissue were observed to evaluate the therapeutic effects on the intestinal damage and inflammatory infiltration. Immunohistochemistry and real-time fluorescent quantitative PCR were employed to determine the expression levels of zonula occludens-1 (ZO-1) and Occludin in the colon tissue, the serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), the expression of Toll-like receptor 4 (TLR4), myeloid differentiation primary response 88 (MyD88), and nuclear factor-κB p65 subunit (NF-κB p65) in the colorectum, and the mRNA levels of TLR4, MyD88, NF-κB p65, and IL-6 in the intestinal tissue. ResultsCompared with the model group, Huaihuasan effectively slowed down the trend of body weight loss (P<0.01), mitigated diarrhea and bloody stool symptoms, reduce the disease activity index (DAI) (P<0.01), restored the length of the colon (P<0.01), and promoted the recovery of the colon tissue. The immunohistochemistry results showed that compared with the model group, Huaihuasan intervention promoted the expression of ZO-1 and Occludin (P<0.05), reduced the serum levels of IL-6 and TNF-α (P<0.05, P<0.01), and down-regulated the protein levels of TLR4, MyD88, and NF-κB p65 in the colon tissue (P<0.05, P<0.01) and the mRNA levels of TLR4, MyD88, NF-κB p65, and downstream IL-6 (P<0.05). ConclusionHuaihuasan exerts significant therapeutic effects on UC due to dampness-heat in the large intestine by modulating the TLR4/NF-κB signaling pathway, reducing inflammatory mediator levels, and enhancing the intestinal barrier function. These findings provide theoretical support for the clinical application and mechanism study of Huaihuasan in the treatment of UC.
6.Effect of Shufeng Jiedu Capsules on Relieving Influenza Virus Pneumonia by Suppressing TLR/NF-κB Pathway in Respiratory Epithelial Cells
Zihan GENG ; Lei BAO ; Shan CAO ; Qiang ZHU ; Jun PAN ; Shuran LI ; Ronghua ZHAO ; Jing SUN ; Yanyan BAO ; Shaoqiu MU ; Xiaolan CUI ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(24):61-68
ObjectiveTo investigate the possible mechanism of Shufeng Jiedu capsules (SFJD) in alleviating influenza A (H1N1) virus pneumonia and focus on its effect on Toll-like receptor (TLR) signaling pathway in respiratory epithelial cells. MethodsA mouse model of viral pneumonia was established via the A/PR/8/34 (PR8) strain of influenza A virus. Mice were randomly divided into a normal group, a PR8 infection (PR8) group, and an SFJD group (8.4 g·kg-1), with 10 mice in each group. The day of infection was designated as day 1. The SFJD group was administered intragastrically at a volume of 20 mL·kg-1 daily, while the normal and PR8 groups were given an equal volume of deionized water. Micro-computed tomography (Micro-CT) was performed on day 5, and the mice were dissected to collect their lungs, after which the lung index was calculated to verify the therapeutic effect of SFJD. Single-cell sequencing was used to analyze the differentially expressed genes in respiratory epithelial cells. Multiplex fluorescence immunohistochemistry was employed to detect the expression of TLR, tumor necrosis factor receptor-associated factor 6 (TRAF6), and myeloid differentiation factor 88 (MyD88) proteins in epithelial cell adhesion molecule (EpCAM)-positive cells, and the proportion of respiratory epithelial cells expressing TLR pathway proteins was calculated. Respiratory epithelial cells were then sorted by flow cytometry, and Western blot was used to detect the expression of TLR, MyD88, TRAF6, Toll-interleukin receptor domain-containing adaptor inducing interferon-β (TRIF), inhibitor of κB kinase α (IKKα), and nuclear factor-κB (NF-κB) in the sorted epithelial cells. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in lung tissue. ResultsAt the transcriptional level, SFJD reversed the expression of TLR signaling pathway genes in respiratory epithelial cells, downregulating multiple TLR signaling pathway-related genes (P<0.01). At the protein level, SFJD significantly reduced the proportion of respiratory epithelial cells expressing TLR3 (P<0.05), the expression levels of TLR2, TLR3, TLR4, TRIF, TRAF6, IKKα, and NF-κB in epithelial cells(P<0.05, P<0.01), as well as the levels of pro-inflammatory cytokines IL-1β and TNF-α in lung tissue (P<0.01). ConclusionSFJD may alleviate viral pneumonia by suppressing the expression of TLR in respiratory epithelial cells and their subsequent signaling cascades.
7.Application research of immersive virtual reality technology in pediatric patients undergoing elective surgery under general anesthesia
Ju ZHANG ; Lili WEI ; Chen XIN ; Jing WANG ; Yan HAN ; Yanyan YANG ; Mengzhu SUN
Chinese Journal of Nursing 2025;60(6):671-676
Objective To explore the application effect of immersive virtual reality(IVR)technology and different viewing content in pediatric patients with general anesthesia during elective surgery,and to provide references for clinical implementation.Methods A total of 180 pediatric patients who underwent elective surgery under general anesthesia in a tertiary hospital in Qingdao from February to October 2023 were selected as study population using convenient sampling method.According to the operation time,60 pediatric patients who underwent surgery from May to July 2023 were included in the immersive panoramic surgical education group,and they could watch the panoramic surgical education video immersively on the basis of routine care.A total of 60 pediatric patients who underwent surgery from August to October 2023 were included in the immersive animation group to watch cartoons immersively on the basis of routine care.A total of 60 pediatric patients who underwent surgery from February to April 2023 were included in the control group and routine preoperative care was implemented.The preoperative anxiety levels,anesthesia induction compliance,and incidence of emergence agitation were compared in the 3 groups by the modified Yale Preoperative Anxiety Scale-Short Form(mYPAS-SF),anesthesia induction cooperation grade,and the Pediatric Anesthesia Emergence Delirium scale(PAED).Results There were statistically significant differences in preoperative anxiety level,anesthesia induced compliance and incidence of emergence agitation during awakening between the 3 groups(P<0.001).Among them,the preoperative anxiety level of the immersive panoramic surgical education group and the immersive animation group was lower than that of the control group,and the difference was statistically significant(P<0.001).The anesthesia induced compliance degree of the immersive panoramic surgical education group and the immersive animation group was better than that of the control group,and the difference was statistically significant(P<0.017),and the incidence of emergence agitation in the immersive panoramic surgical education group was lower than that of the control group,and the difference was statistically significant(P=0.004).Conclusion The use of IVR technology to watch panoramic surgical education videos and cartoons can help reduce the preoperative anxiety level and improve anesthesia induction cooperation degree of pediatric patients with general anesthesia during elective surgery,but the intervention effect of panoramic surgical education videos is better in improving the emergence agitation.
8.Elderly acute progressive cerebral infarction patients' serum PLA2,called S100B levels and the relation-ship between carotid plaques
Yanyan JIA ; Yijia SUN ; Di ZHAO ; Yan SONG
The Journal of Practical Medicine 2025;41(2):244-249
Objective To explore the relationship between serum phospholipase A2(PLA2),S100 calcium-binding protein B(S100B) and carotid plaque in elderly patients with acute progressive cerebral infarction. Methods From January 2021 to June 2022,120 elderly patients with acute progressive cerebral infarction were enrolled in the study group,while 115 patients with non-progressive cerebral infarction were classified as non-progressive group and 100 healthy people as healthy group. The study group was divided into three groups:no plaque group (34 cases),stable plaque group (45 cases) and unstable plaque group (41 cases). To analyze the relationship between serum PLA2 and S100B levels and carotid plaque. ROC analysis of their predictive value for carotid plaque. Follow-up for 1 year,the survival rate was analyzed with recurrence and death as the end points. Results The levels of serum PLA2 and S100B in stable and unstable plaque groups were higher than those in non-plaque group,and the difference was statistically significant (P<0.05). Serum PLA2 and S100B were positively correlated with carotid plaque. ROC showed that the value of joint prediction was higher (P<0.05). Conclusion The levels of serum PLA2 and S100B are increased in elderly patients with acute progressive cerebral infarction,which is closely related to carotid plaque.
9.Effects of exercise during pregnancy on renal fibrosis and Ang Ⅱ/TGF-β1/CTGF sig-naling pathway in offspring of spontaneously hypertensive rats
Shuang LIU ; Teng GAO ; Hualing SUN ; Yanyan ZHANG ; Lijun SHI
Chinese Journal of Arteriosclerosis 2025;33(1):16-23
Aim To explore the effects of exercise during pregnancy on renal structure,function and angiotensin Ⅱ(Ang Ⅱ)/transforming growth factor-β1(TGF-β1)/connective tissue growth factor(CTGF)signaling pathway in 3-month-old offspring of spontaneously hypertensive rats(SHR),the aim of this study was to provide experimental basis for early intervention of hypertension and protection of key target organs.Methods After mating SHR and WKY rats,pregnant rats were randomly divided into sedentary group(p-WKY-SED,p-SHR-SED)and exercise group(p-WKY-EX,p-SHR-EX).Blood pressure,serum urea nitrogen and creatinine were measured by caudal artery non-invasive blood pressure system and colorimetry in 3-month-old offspring rats.HE staining,Masson staining,ELISA and Western blot were used to detect the renal structure,collagen volume fraction,Ang Ⅱ concentration,renin-angiotension-aldosterone sys-tem(RAAS)and protein expression related to fibrogenic signal pathway in 3-month-old rats.Results(1)The sys-tolic blood pressure(SBP),diastolic blood pressure(DBP)and mean arterial pressure(MAP)of offspring rats in p-SHR-SED group were significantly higher than those in p-WKY-SED group.The SBP,DBP and MAP of SHR male off-spring rats were significantly decreased by exercise during pregnancy(P<0.05),but had no effect on the female offspring rats(P>0.05).(2)There was no significant difference in serum urea nitrogen and creatinine among the groups(P>0.05).(3)The glomerular volume and the collagen volume fraction in p-SHR-SED group were significantly higher than those in p-WKY-SED group(P<0.05),and the glomerular volume and the collagen volume fraction in p-SHR-EX group were significantly lower than those in p-SHR-SED group(P<0.05).(4)Renal Ang Ⅱ level of offspring rats in p-SHR-SED group was significantly higher than that in p-WKY-SED group,and renal Ang Ⅱ level of offspring rats in p-SHR-EX group was significantly lower than that in p-SHR-SED group(P<0.05).(5)The expression levels of angiotensin Ⅱ type 1 receptor(AT1R),TGF-β1 and CTGF protein in p-SHR-SED group were significantly higher than those in p-WKY-SED group(P<0.05),while the expression levels of angiotensin converting enzyme 2(ACE2),angiotensin Ⅱ type 2 receptor(AT2R)and MasR protein in p-SHR-SED group were significantly lower than those in p-WKY-SED group(P<0.05).Conclusion(1)Exercise during pregnancy can significantly decrease the blood pressure of 3-month-old male offspring rats of hypertensive rats,but has no significant effect on that of 3-month-old female offspring.(2)Exercise during preg-nancy may reduce renal fibrosis in 3-month-old female/male offspring of hypertensive rats by regulating RAAS balance and inhibiting Ang Ⅱ/TGF-β1/CTGF signaling pathway.
10.Cloning and expression of PPE65 encoded by the Mycobacterium tuberculosis Rv3621c gene in Escherichia coli,and its effects on proliferation and TGF-β expression of BEAS-2B cells
Juncheng HUANG ; Xinwen BO ; Jing LI ; Xuke CHEN ; Jiaxin ZHAO ; Yanyan ZHANG ; Xinwei FENG ; Yan SUN ; Zhengrong WANG ; Meng QI
Chinese Journal of Zoonoses 2025;41(10):1025-1033
This study was aimed at preliminarily investigating the molecular biological functions of the PPE65 protein from Myco-bacterium tuberculosis,and providing foundational data for tuberculosis prevention and control.The basic biological properties of the PPE65 gene-encoded protein were predicted with bioinformatics tools.Sequence information for the Mycobacterium tuberculosis Rv3621c gene and PPE65 protein was retrieved from the NCBI database.The Rv3621c gene was amplified through PCR with the H37Rv genome as a template,then cloned into the pET22b(+)expression vector.The recombinant pET22b(+)-PPE65 plasmid was transformed into Escherichia coli BL21(DE3)competent cells for IPTG-induced expression.Solubility analysis,purification,and identification of the recombinant PPE65 protein were performed.BEAS-2B cells were treated with various concentrations of PPE65 protein for 24 h,and cell proliferation was assessed with CCK-8 assays.PPE65 was found to be composed of 413 amino acids and to have a molecular formula of C????H????N???O???S??,a relative molecular mass of 40 679.88,a theoretical isoelectric point of 4.60,an ali-phatic index of 81.94,and an average hydrophilicity value of 0.319,thus indicating a stable hydrophobic protein lacking signal pep-tides or transmembrane domains.Secondary structure analysis revealed 53.03%α-helix(Hh),2.66%β-sheet(Ee),and 44.31%ran-dom coil(Cc).Bioinformatics predictions identified 38 B-cell epitopes and 22 CTL/Th-cell epitopes.The full-length PPE65 gene(1 308 bp)was confirmed through double restriction enzyme digestion and sequencing,thereby validating the correct construction of the pET22b(+)-PPE65 recombinant plasmid.SDS-PAGE analysis demonstrated that the recombinant protein was found in inclusion bodies,and a single band at 43.7 kDa was observed after purification.Western blotting revealed specific binding to mouse-derived His monoclonal antibodies,thereby confirming successful expression of the PPE65 protein.BEAS-2B cells treated with a PPE65 protein concentration gradient(2.5-20 μg/mL)exhibited a dose-dependent increase in cell number.Compared with those in the PBS control group,TGF-β relative expression levels were significantly higher in all treatment groups(t2.5=4.893,P<0.001,t5.0=4.640,P<0.05,t10=7.535,P<0.05,t20=16.44,P<0.000 1).This study elucidated the structural characteristics of the PPE65 protein,successfully obtained the recombinant protein through prokaryotic expression and purification,and demonstrated its ability to promote BEAS-2B cell proliferation.The underlying mechanism might involve suppression of TGF-β/S mad signaling pathway activation.These findings provide a theoretical basis for understanding the role and regulatory mechanisms of PPE65 during M.tuberculosis infection.

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