1.Diagnosis and differential diagnosis of mucin-rich salivary gland tumors
GUAN Weihang ; LIU Cangwei ; GUO Hao ; LI Jinwei ; WANG Dandan ; QIAO Chunyan ; NIE Mengdong ; QU Ming ; SHI Ce
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(6):606-619
This paper systematically elaborates on the key points of diagnosis and differential diagnosis of salivary gland tumors characterized by a substantial amount of extracellular mucus as a main or prominent feature, and clarifies the core differential features. The term "mucus-rich" specifically denotes that mucus is a major component of the tumor, rather than a focal or minor one. This phenomenon is associated with distinct histogenetic mechanisms: it may result from specific genetic mutations (e.g., AKT1 E17K in mucinous adenocarcinoma) that drive ductal epithelial differentiation into mucus-secreting cells, or from myoepithelial cells secreting glycosaminoglycans that form a myxoid stroma. Salivary gland tumors with abundant extracellular mucus include mucinous cystadenoma, sialadenoma papilliferum-like intraductal papillary tumors, mucinous myoepithelioma, pleomorphic adenoma with mucin-rich stroma, mucinous adenocarcinoma, low-grade mucoepidermoid carcinoma, mucin-rich salivary duct carcinoma and intestinal-type adenocarcinoma. The diagnosis of these tumors is complicated by the dual nature of extracellular mucus: while it is a defining feature of some entities, it can also obscure key diagnostic architectural features in others, leading to histological overlap and inconspicuous diagnostic areas. Given the frequent histological morphological overlap among these tumors, immunohistochemical findings and molecular characteristics have emerged as crucial differential diagnostic criteria. Core differential diagnostic points include the following: histologically, there must be meticulous identification of typical structures obscured by mucin (such as squamoid cells in mucoepidermoid carcinoma and apocrine features in salivary duct carcinoma); in immunohistochemical staining, CK20 is useful for distinguishing intestinal-type adenocarcinoma (positive) from mucinous adenocarcinoma (negative), while androgen receptor aids in differentiating salivary duct carcinoma (positive) from mucoepidermoid carcinoma (negative); and molecular testing plays a critical role in definitive diagnosis (e.g., the AKT1 E17K mutation for mucinous adenocarcinoma, MAML2 rearrangement for mucoepidermoid carcinoma, and MEF2C::SS18 fusion for microsecretory adenocarcinoma). This paper systematically summarizes the core pathological features and differential diagnostic points of mucin-rich salivary gland tumors, aiming to provide a practical reference for clinical pathological diagnosis.
2.The Specificity of Electroacupuncture at Different Acupoints in Promoting Cerebrospinal Fluid Flow in Mice
Yu SHI ; Qian HUA ; Tian-Tian PENG ; Yu-Xin NIE ; Zhao-Heng LIU ; Chen-Geng DENG ; Xu WANG
Progress in Biochemistry and Biophysics 2026;53(5):1154-1164
ObjectiveCerebrospinal fluid (CSF) plays a crucial role in maintaining the homeostasis of the central nervous system (CNS). CSF rapidly exchanges with interstitial fluid (ISF) via the glymphatic system within the brain parenchyma. CSF-ISF circulation and its associated mechanisms are often referred to as the brain lymphatic system. This system is connected directly to meningeal lymphatic vessels (mLVs), jointly performing the function of clearing metabolic waste from the CNS. Emerging evidence indicates that this system is closely associated with the onset and progression of neurodegenerative diseases (NDs) such as Alzheimer’s disease (AD). Importantly, abnormal CSF circulation is not only a downstream consequence of AD pathology, but also a risk factor. In AD, the dynamics of CSF flow within the CNS are diminished, immune dysregulation occurs, and this may increase the risk of AD by exacerbating the burden of amyloid β-protein (Aβ). In the mouse model of AD, impaired CSF flow compromises this clearance function, leading to cognitive deficits. Clinically, acupuncture at cognition-related acupoints is commonly used for the prevention and treatment of AD. However, whether its therapeutic effects are mediated through the modulation of CSF dynamics remains unclear. This study aimed to evaluate the impact of acupuncture on CSF flow and investigate its acupoint specificity. MethodsMice were randomly assigned to experimental groups for the different electroacupuncture groups with the following acupoints: Baihui point (GV 20), Ear point, Neiguan point (PC 6), and Tianshu point (ST 25). Wild-type mice on a C57BL/6J background were used as controls. Fluorescent tracer was injected into the cisterna magna to label CSF flow. Fluorescence imaging was employed to assess the distribution of CSF within the brain before and after acupuncture stimulation. ResultsFollowing tracer injection into the cisterna magna, fluorescence signals rapidly reached the cerebellum and medulla—the regions closest to the injection site. Fluorescence intensity was higher in ventral brain regions compared to dorsal regions, likely due to greater vascular density in ventral areas facilitating CSF-ISF exchange. Electroacupuncture at the GV 20 produced the most pronounced enhancement of CSF across the whole brain, while stimulation at the ST 25 primarily augmented flow within subcortical regions. In contrast, electroacupuncture at the Ear point or the PC 6 had no observable effect on CSF in mice. ConclusionElectroacupuncture promotes CSF flow into the brain parenchyma in an acupoint-specific manner, with GV 20 exhibiting the most pronounced enhancement of CSF dynamics. These findings suggest that acupuncture-mediated facilitation of CSF flow may represent a potential therapeutic strategy for preventing or delaying age-related cognitive decline.
3.The Specificity of Electroacupuncture at Different Acupoints in Promoting Cerebrospinal Fluid Flow in Mice
Yu SHI ; Qian HUA ; Tian-Tian PENG ; Yu-Xin NIE ; Zhao-Heng LIU ; Chen-Geng DENG ; Xu WANG
Progress in Biochemistry and Biophysics 2026;53(5):1154-1164
ObjectiveCerebrospinal fluid (CSF) plays a crucial role in maintaining the homeostasis of the central nervous system (CNS). CSF rapidly exchanges with interstitial fluid (ISF) via the glymphatic system within the brain parenchyma. CSF-ISF circulation and its associated mechanisms are often referred to as the brain lymphatic system. This system is connected directly to meningeal lymphatic vessels (mLVs), jointly performing the function of clearing metabolic waste from the CNS. Emerging evidence indicates that this system is closely associated with the onset and progression of neurodegenerative diseases (NDs) such as Alzheimer’s disease (AD). Importantly, abnormal CSF circulation is not only a downstream consequence of AD pathology, but also a risk factor. In AD, the dynamics of CSF flow within the CNS are diminished, immune dysregulation occurs, and this may increase the risk of AD by exacerbating the burden of amyloid β-protein (Aβ). In the mouse model of AD, impaired CSF flow compromises this clearance function, leading to cognitive deficits. Clinically, acupuncture at cognition-related acupoints is commonly used for the prevention and treatment of AD. However, whether its therapeutic effects are mediated through the modulation of CSF dynamics remains unclear. This study aimed to evaluate the impact of acupuncture on CSF flow and investigate its acupoint specificity. MethodsMice were randomly assigned to experimental groups for the different electroacupuncture groups with the following acupoints: Baihui point (GV 20), Ear point, Neiguan point (PC 6), and Tianshu point (ST 25). Wild-type mice on a C57BL/6J background were used as controls. Fluorescent tracer was injected into the cisterna magna to label CSF flow. Fluorescence imaging was employed to assess the distribution of CSF within the brain before and after acupuncture stimulation. ResultsFollowing tracer injection into the cisterna magna, fluorescence signals rapidly reached the cerebellum and medulla—the regions closest to the injection site. Fluorescence intensity was higher in ventral brain regions compared to dorsal regions, likely due to greater vascular density in ventral areas facilitating CSF-ISF exchange. Electroacupuncture at the GV 20 produced the most pronounced enhancement of CSF across the whole brain, while stimulation at the ST 25 primarily augmented flow within subcortical regions. In contrast, electroacupuncture at the Ear point or the PC 6 had no observable effect on CSF in mice. ConclusionElectroacupuncture promotes CSF flow into the brain parenchyma in an acupoint-specific manner, with GV 20 exhibiting the most pronounced enhancement of CSF dynamics. These findings suggest that acupuncture-mediated facilitation of CSF flow may represent a potential therapeutic strategy for preventing or delaying age-related cognitive decline.
4.Treatment Modalities and Long-Term Outcomes in Unruptured Vertebrobasilar Fusiform Aneurysms: A Nationwide Observational Cohort Study
Linggen DONG ; Dachao WEI ; Xiheng CHEN ; Mingtao LI ; Yang ZHAO ; Yong SUN ; Qingbin NIE ; Jun FENG ; Guomin XIAO ; Jinghua ZHOU ; Shengli HU ; Lifei FENG ; Lifeng QI ; Hongen LIU ; Geng GUO ; Yufang LI ; Renfu TIAN ; Jianghua YU ; Dianshi JIN ; Liang HAO ; Tian TIAN ; Shizhong ZHANG ; Yang WANG ; Liping LIU ; Ming LV
Journal of Stroke 2026;28(2):250-262
Background:
and Purpose Vertebrobasilar fusiform aneurysms (VBFAs) carry substantial morbidity and mortality, but optimal management for unruptured VBFAs remains unclear. We compared the safety and efficacy of conservative management (CM), stent-assisted coiling (SAC), and flow diverters (FDs) in patients with unruptured VBFAs, focusing on long-term prognosis.
Methods:
This study included data from a nationwide Chinese cohort of patients with vertebrobasilar dissecting aneurysms. Inverse probability of treatment weighting (IPTW) balanced confounders across groups. The primary outcome was poor prognosis (modified Rankin Scale score >2). Secondary outcomes included aneurysm rupture, ischemic stroke, compression symptoms, and VBFA-related deaths. Logistic regression estimated odds ratios (ORs) and 95% confidence intervals (CIs). Subgroup and sensitivity analyses were performed.
Results:
Among 1,115 patients with unruptured VBFAs, 838 (median age, 54 years; 655 men) were included. After IPTW, baseline characteristics were balanced. Median follow-up was 54 months. FD was associated with a lower risk of poor prognosis than CM (OR, 0.48 [95% CI, 0.30 to 0.77]; p=0.002), with no difference between CM and SAC. FD also reduced aneurysm rupture (OR, 0.20 [95% CI, 0.07 to 0.60]; p=0.004) and compression symptoms (OR, 0.30 [95% CI, 0.13 to 0.68]; p=0.004) versus CM. Time-to-event analyses further revealed significant differences in vertebral artery lesions and Type I–II VBFAs, whereas no significant differences were observed in basilar or vertebrobasilar junction lesions or in Type III–IV VBFAs.
Conclusions
Compared with CM, FD was associated with improved long-term outcomes in unruptured VBFAs, particularly in vertebral artery lesions and Type I–II VBFAs, although residual confounding cannot be excluded.
5.Generation of Cynomolgus and Rhesus Monkey Models Carrying the BMPR1BFecB Variant
Chunyang LI ; Xingchen LIU ; Yuzhuo LI ; Yong LU ; Yan WANG ; Changshan GAO ; Yanhong NIE ; Xinyan BIAN ; Chao HU ; Jiaqiao ZHU ; Qiang SUN
Laboratory Animal and Comparative Medicine 2026;46(4):467-475
ObjectiveTo generate cynomolgus and rhesus monkey models carrying the Booroola fecundity (FecB) variant of bone morphogenetic protein receptor type 1B (BMPR1B) gene (designated BMPR1BFecB) using adenine base edit technology and to explore genetic improvement strategies for enhancing the reproductive performance of laboratory macaques. MethodsFour single-guide RNAs (sgRNAs) were designed to target the BMPR1B c.746A>G locus in cynomolgus and rhesus monkeys. The sgRNAs were combined with the ABE8e-SpRY base editor, and the resulting editing complexes were microinjected into zygotes of both macaque species. Following in vitro culture of the injected embryos, nested PCR amplification was performed to detect the base-editing efficiency at the target site to screen for highly efficient sgRNAs. Subsequently, normally developing gene-edited embryos were transferred into the oviducts of surrogates. Seven days after birth, ear skin or peripheral blood samples were collected from the newborn monkeys for genotyping and mosaicism analysis, and the growth, development, and health status of the gene-edited monkeys were monitored. ResultsAmong the four candidate sgRNAs, sgRNA4 was identified as achieving 100% editing efficiency at the c.746A>G site. A total of 17 live monkey offspring were obtained via embryo transfer, including 8 cynomolgus monkeys and 9 rhesus monkeys. Genotyping results showed that all 8 newborn cynomolgus monkeys carried the c.746A>G point mutation, with an editing efficiency of 100% (8/8), of which 37.50% (3/8) were mosaics. Among the 9 newborn rhesus monkeys, 8 individuals carried the c.746A>G mutation, with an editing efficiency of 88.89% (8/9), of which 75.00% (6/8) were mosaics. ConclusionThe ABE8e-SpRY base-editing system enables precise editing at the c.746A>G (p.Q249R) locus of the BMPR1B gene in both cynomolgus and rhesus monkey embryos, resulting in the successful establishment of laboratory monkey models carrying the BMPR1BFecB variant. These models provide a valuable foundation for future studies of the regulatory role of the BMPR1BFecB variant in reproductive performance at the non‑human primate level, as well as for exploring genetic improvement strategies to enhance reproductive performance in laboratory monkeys.
6.Effects of Electrothermal Acupuncture on the HIF-1α/S1P/S1PR3 Pathway and Macrophage Polarization in Epididymal White Adipose Tissue of Obese Model Mice
Xiaobo GE ; Yuanyuan LI ; Jiangyan WEI ; Limin NIE ; Xin YANG ; Quan MIAO ; Mina WANG ; Yawei XI ; Luopeng ZHAO ; Yine SONG ; Xinru SUN ; Lu LIU ; Bin LI
Journal of Traditional Chinese Medicine 2026;67(20):2227-2235
ObjectiveTo investigate the mechanism underlying the weight loss effect of electrothermal acupuncture (ETA) in diet-induced obese (DIO) model mice based on the hypoxia-inducible factor-1α (HIF-1α)/sphingosine-1-phosphate (S1P)sphingosine-1-phosphate receptor 3 (S1PR3) signaling pathway-mediated macrophage polarization. MethodsAn obesity mouse model was established by continuously feeding at a high-fat diet (HFD) for 8 weeks. Twenty successfully modeled obese mice were randomly divided into a model group and an electrothermal acupuncture (ETA) group with 10 mice per group, both of which continued to receive the HFD till week 12. Another 10 mice were assigned to the normal group and fed a standard chow diet continuously for 12 weeks. Starting from week 9 of the experiment, mice in the ETA group received electrothermal acupuncture stimulation at Zhongwan (CV12), bilateral Tianshu (ST25), and bilateral Zusanli (ST36) for 30 minutes per session, with the instrument current set at (40±0.5)mA and a constant temperature of (40±0.5)℃. Mice in the normal group and the model group were restrained in the same manner for 30 minutes without acupuncture. All interventions were performed once every other day for a total of 4 weeks. Ten hours after the final intervention, the body weight, waist circumference, and body length of mice in each group were measured, and Lee's index was calculated. Serum biochemical analysis was performed to assess fasting blood glucose (FBG), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (Cr). Serum leptin (LEP) and adiponectin (ADPN) levels, as well as sphingosine-1-phosphate (S1P) levels in epididymal white adipose tissue (eWAT), were detected by ELISA. Histomorphological changes in eWAT were monitored via HE staining. Flow cytometry was used to analyze macrophage subtype-related indexes in eWAT, including the total proportion of macrophages, the proportion of M1 macrophages, the proportion of M2 macrophages, and the M1/M2 ratio. Antibody array technology was used to determine the levels of differentially expressed inflammatory factors in eWAT between the model and blank groups, as well as between the ETA and model groups. Western blot and qPCR were used to detect the protein and mRNA expression levels of HIF-1α and S1PR3 in the eWAT, respectively. ResultsCompared to the normal group, the model group showed significantly increased body weight, Lee's index, waist circumference, and FBG, TC, TG, HDL-C, LDL-C, AST, and ALT levels, upgraded LEP and S1P levels, decreased ADPN level and ADPN/LEP ratio, elevated proportion of M1 macrophages and the M1/M2 ratio, reduced proportion of M2 macrophages, and upregulated protein and mRNA expression levels of HIF-1α and S1PR3 (P<0.05 or P<0.01). With the exception of HDL-C, all the abovementioned indicators in the ETA group were significantly improved compared to the model group (P<0.05 or P<0.01). Analysis of differentially expressed inflammatory cytokines revealed that, compared to the normal group, the levels of interleukin-1β (IL-1β), interferon-γ (IFN-γ), interleukin-6 (IL-6), and interleukin-12p70 (IL-12p70) in the eWAT of the model group were elevated; conversely, the levels of interleukin-17 (IL-17), interleukin-5 (IL-5), IL-6, and interleukin-23 (IL-23) in the eWAT of the ETA group decreased compared to the model group (P<0.05 or P<0.01). ConclusionETA exerts a weight-reducing effect in obese model mice, and the mechanism may involve inhibition of the HIF-1α/S1P/S1PR3 signaling pathway activation, which shifts macrophage polarization from the M1 to M2 phenotype, thereby alleviating chronic inflammation.
7.The preventive and therapeutic effects of three ethanol extracts derived from three sources of Stellera chamaejasme L. on imiquimod-induced psoriasis in mice
Ying PENG ; Xin LIU ; Yiwen NIE ; Xinhe WANG ; Hua NIAN ; Jianyong ZHU
Journal of Pharmaceutical Practice and Service 2026;44(10):502-507
Objective To investigate the preventive and therapeutic effects of ethanol extracts derived from three sources of traditional Chinese medicine: Stellera chamaejasme L., Euphorbia fischeriana Steud. and Euphorbia kansuensis Prokh., on imiquimod (IMQ)-induced psoriasis in mice. Methods Thirty-six male BALB/c mice were randomly divided into the following 6 groups with 6 mice in each group: blank control, model, Stellera chamaejasme, Euphorbia fischeriana, Euphorbia kansuensis, and calcipotriol. Psoriasis area and severity index(PASI) scores were used to record the changes of skin lesions in every group; Hematoxylin-eosin(HE) staining was used to observe the pathological morphology of skin and measure the thickness of the epidermis. Immunohistochemistry was used to detect the expression of nuclear antigen Ki67 in the skin tissues of mice. Results Compared with the model group, the three kinds of ethanol extracts could reduce the PASI score, inhibit epidermal thickening, and decrease expression of Ki67 in the psoriasis mice. Among them, the therapeutic effect of Stellera chamaejasme was the most significant and it was better than the commonly used topical drug calcipotriol. Conclusion The ethanol extract of Stellera chamaejasme had good anti-psoriatic activity, which could inhibit the abnormal proliferation of keratinocytes, reduce the expression of Ki67, and significantly improve psoriasis-like skin lesions.
8.Construction and practice of "three-stage" teaching empowers the ideological and political teaching model in the introduction to nursing course
Ying ZHU ; Yuhui ZHU ; Zhenli WANG ; Fangfang LIU ; Qingqing NIE ; Yating WANG ; Leshan ZHOU
Chinese Journal of Practical Nursing 2025;41(13):1025-1029
Objective:To explore the ideological and political education elements and methods in the introduction to nursing course, so as to enhance its ideological educational effectiveness. Methods:In February 2024, based on a "three-stage" teaching model (pre-class, in-class, and post-class), this study reformed the ideological and political education plan for the introduction to nursing course by integrating online and offline case studies and resources, expanding educational materials, and organizing teaching assessments and effectiveness surveys. A semester-long ideological and political education practice was implemented for 237 first-year nursing students from four classes in Medical College of Xinjiang University of Science and Technology. The effectiveness was evaluated by exam scores and a self-designed Introduction to Nursing Teaching Effectiveness Evaluation Scale under the ideological and political education framework. Results:Among the 237 students, there were 107 males and 130 females, aged (18.50 ± 1.12) years old, 63 achieved a passing grade, 149 achieved a good grade, and 18 achieved an excellent grade, with a pass rate of 97.05% (230/237). The scores of the six dimensions of interest stimulation in teaching methods, acceptance of teaching methods, knowledge review and application, knowledge comprehension and retention, self-challenge and improvement, and satisfaction with learning outcomes within 1 week after the end of the course scores were (3.92 ± 1.11), (4.21 ± 0.82), (4.05 ± 0.97), (4.05 ± 0.89), (4.11 ± 0.94), (4.03 ± 0.90) points, which were significantly higher than 1 week before the start of the course (3.43 ± 1.00), (3.54 ± 1.03), (3.48 ± 1.08), (3.47 ± 1.04), (3.52 ± 1.05), (3.47 ± 1.01) points. The differences were statistically significant ( t values were -19.38- -13.22, all P < 0.01). Conclusions:The "three-stage" teaching model in the introduction to nursing course effectively integrates professional education with ideological and political education, improves students′ academic performance, enhances learning effectiveness in nursing practice, and strengthens the overall impact of ideological and political education.
9.Comparison of bladder volume measurement accuracy between two-dimensional ultrasound with three-dimensional reconstruction and conventional two-dimensional ultrasound
Kaixuan ZHANG ; Ying CAO ; Lijing ZUO ; Zhen WANG ; Wensheng NIE ; Yongli SONG ; Xing LIU ; Mingjian SUN ; Yuan TANG ; Yueping LIU
Chinese Journal of Radiation Oncology 2025;34(12):1238-1244
Objective:To compare the accuracy of two-dimensional (2D) ultrasound with three-dimensional (3D) reconstruction and conventional 2D ultrasound in measuring bladder volume in pelvic tumor patients, using computed tomography (CT) as the reference.Methods:A set of bladder phantoms were constructed to compare CT and ultrasound measurements with actual injected volumes. Clinical data of 104 pelvic tumor patients who received radiotherapy at the Cancer Hospital, Chinese Academy of Medical Sciences between August and December 2023 were retrospectively analyzed. Portable transabdominal ultrasound was used to obtain the largest bladder cross-section, and the maximum diameters in the left-right (LR), anterior-posterior (AP), and superior-inferior (SI) directions (D LR, D AP, D SI) were measured. The 2D ultrasound volume was calculated as V=0.523 × D LR × D AP × D SI. Full-bladder transverse videos were recorded and processed in Matlab R2016a through frame extraction(60 images), followed by contrast enhancement, edge detection segmentation, cubic spline interpolation, and image smoothing to achieve 3D reconstruction. Paired t-tests, intraclass correlation coefficients (ICC), and Bland-Altman analyses were performed to assess systematic bias and consistency between ultrasound methods and CT. Multivariate linear regression was applied to evaluate the effects of slice thickness, posture, age, and other factors on CT measurements. Results:In the phantom study, deviations of 2D ultrasound and CT from actual injected volumes were (0.73±3.05) ml ( t=-0.48, P=0.667) and (1.52±11.27) ml ( t=0.17, P=0.875), with ICC values>0.999. In the clinical study, mean bladder volumes measured by 3D-reconstructed ultrasound, conventional 2D ultrasound, and CT were (373.5±153.31), (314.89±135.28), (382.82±157.57) ml, respectively. The 3D-reconstructed method showed excellent agreement with CT (ICC=0.98; Bland-Altman mean bias=-9.32 ml, P=0.096), while 2D ultrasound also showed good consistency (ICC=0.91), but significantly underestimated bladder volume (mean bias=-67.93 ml, P<0.001). Subgroup analysis revealed that 2D ultrasound had the best agreement with CT in the medium-volume group (200-500 ml, ICC=0.902), whereas agreement decreased in the small-volume (<200 ml, ICC=0.884) and large-volume (>500 ml, ICC=0.840) groups (all P<0.001). The 3D-reconstructed ultrasound maintained excellent consistency with CT across all subgroups (all ICC>0.95), and the measured bladder volume was not statistically significant. Multivariate regression showed that slice thickness, posture, age, sex, and surgical status had no significant effects on CT measurements. Conclusions:Ultrasound with 3D reconstruction enables accurate bladder volume monitoring through true 3D contour reconstruction, while conventional 2D ultrasound systematically underestimates bladder volume and requires correction.
10.PE_PGRS37 protein promotes intracellular colonization of Mycobacterium tuberculosis by inhibiting macrophage autophagy flow
Mengyu LI ; Zhenjun ZHANG ; Tingting FENG ; Hui WANG ; Chanchan NIE ; Chunwen CHEN ; Yunjie GAO ; Yifan DUAN ; Ruonan GUO ; Yingying CUI ; Guanghui DANG ; Siguo LIU
Chinese Journal of Zoonoses 2025;41(10):1005-1010,1015
This study explored the effect of the Mycobacterium tuberculosis(Mtb)PE/PPE family protein PE_PGRS37 on the growth of Mycobacterium smegmatis(Ms)and macrophage autophagy during Mtb infection.The pe_pgrs37 gene was amplified from Mtb genome through PCR,and the recombinant vector pAIN-PE_PGRS37 was successfully constructed through homologous recombi-nation.pAIN-PE_PGRS37 and pAIN were integrated into Ms through electroshock to construct pAIN-PGRS37/Ms and pAIN/Ms re-combinant bacteria.Western blotting indicated that the PE_PGRS37 protein was correctly expressed in pAIN-PE_PGRS37/Ms.The re-combinant bacteria were inoculated in 7H9/7H10 medium,and their colony morphology and growth curves were observed.No signifi-cant difference in colony morphology was observed between pAIN-PE_PGRS37/Ms and pAIN/Ms.The growth rate significantly in-creased between 10 and 16 h,and a plateau was reached at 26 h.After infection of U937 cells with pAIN-PE_PGRS37/Ms and pAIN/Ms,macrophage autophagy flow was detected with western blotting and immunofluorescence.In the pAIN-PE_PGRS37/Ms-infected group,compared with the pAIN/Ms-infected group,macrophage LC3-II and p62 protein expression was significantly up-regulated(P<0.001)and inhibited autophagosome and lysosome fusion.The intracellular survival of the recombinant bacteria was detected through colony counting,and pAIN-PE_PGRS37/Ms showed significantly greater survival in macrophages at 12 h,24 h,and 48 h than pAIN/Ms(P<0.05).Our results suggested that PE_PGRS37 protein promotes Mycobacterium survival in macrophages by blocking macro-phage autophagy flow,thus inhibiting macrophage autophagy.


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