1.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.
2.The efficacy of individualized endovascular interventional treatment for extracranial carotid artery pseudoaneurysms
Tengfei LI ; Chengcheng SHI ; Sun YU ; Ji MA ; Ling WANG ; Quanhui ZHANG ; Xinwei HAN
Chinese Journal of Radiology 2025;59(6):712-718
Objective:To investigate the efficacy and safety of different endovascular interventional treatments for extracranial carotid artery pseudoaneurysms.Method:The clinical data of 48 patients with extracranial carotid artery pseudoaneurysms treated with multiple endovascular procedures were retrospectively analyzed in the First Affiliated Hospital of Zhengzhou University from February 2012 to February 2024. The patients presented with a total of 48 extracranial carotid pseudoaneurysms, ranging in diameter from 2.5 to 34.2 mm [mean (12.0±9.6) mm]. The lesions were distributed as follows: 25 in the internal carotid artery, 16 in the external carotid artery and 7 in the common carotid artery. The selection of endovascular interventional techniques was tailored to individual cases according to the pseudoaneurysm size, anatomical location, morphological configuration, and specific features of the parent artery. Perioperative adverse events were monitored, and the efficacy of individualized endovascular interventional therapy was evaluated based on immediate postoperative and 6-month follow-up digital subtraction angiography (DSA) findings, including aneurysm occlusion and in-stent patency.Result:The treatment modalities included parent artery coiling occlusion ( n=16), overlapping braided carotid stent implantation ( n=3), covered stent placement ( n=23), combined implantation of covered stent and braided carotid stent implantation ( n=4) and flow-diverting stent implantation ( n=2) based on the characteristics of the pseudoaneurysms. Endovascular interventional procedures were successfully completed in 47 patients (technical success rate: 97.9%). Immediate postoperative DSA revealed residual pseudoaneurysm at the distal end of the stent in 1 case. Among the remaining cases, complete aneurysm obliteration or faint opacification was observed, with stent lumen patency confirmed in 31 cases and complete parent artery occlusion achieved in 16 cases. In one case involving a patient who underwent flow-diverting stent implantation, a pulsatile vascular murmur reappeared in the neck one week postoperatively. Follow-up DSA revealed stent migration into the aneurysm sac. After retrieval and removal of the displaced stent, combined implantation of a covered stent and a braided carotid artery stent was performed. Postoperative angiography confirmed complete aneurysm occlusion and patent parent artery blood flow. No severe perioperative adverse events (e.g., aneurysm rupture) were observed. During a postoperative follow-up period of 6.2-24.2 months, DSA at 6 months revealed mild in-stent or distal segment stenosis in 2 patients who underwent covered stent implantation. The remaining 46 patients exhibited complete aneurysm occlusion with no significant stenosis observed within the stent lumen. At the final follow-up, all patients demonstrated resolution or significant alleviation of clinical symptoms. Conclusion:Individualized endovascular interventional therapy demonstrates favorable safety and efficacy profiles in managing extracranial carotid artery pseudoaneurysms.
3.A multicenter study evaluating the efficacy of bronchial artery chemoembolization combined with anlotinib for advanced non-small cell lung cancer
Chao LIANG ; Hao LI ; Donglin KUANG ; Daqian HAN ; Jiacheng WANG ; Yanji ZHANG ; Yifan ZHAI ; Mengkun LIU ; Huibin LU ; Dechao JIAO ; Jianzhuang REN ; Shenghai LIANG ; Chenguang PANG ; Shiqi ZHOU ; Yanliang LI ; Xinwei HAN ; Yong WANG ; Xuhua DUAN
Chinese Journal of Radiology 2025;59(11):1293-1301
Objective:To compare the clinical efficacy and safety of bronchial artery chemoembolization (BACE) combined with anlotinib (BACE+A) versus BACE alone in patients with stage III-IV non-small cell lung cancer (NSCLC).Methods:A total of 94 patients with advanced NSCLC treated at six interventional centers between November 2020 and November 2021 were retrospectively enrolled. Patients were divided into the BACE+A group ( n=46) and the BACE alone group ( n=48) based on treatment regimen. Baseline and perioperative clinical data were collected and compared between the two groups. Treatment response was evaluated using the modified Response Evaluation Criteria in Solid Tumors (mRECIST) at 1, 6, and 12 months after the first BACE procedure. Objective response rate (ORR), disease control rate (DCR), progression-free survival (PFS), overall survival (OS), and treatment-related adverse events (AEs) were recorded. Kaplan-Meier survival curves were plotted to compare median OS and PFS between groups. Cox proportional hazards regression analysis was used to identify factors influencing OS and PFS. Results:The Kaplan-Meier analysis showed that the median OS was significantly longer in the BACE+A group (18.8 months, 95% CI 16.3-21.3) than in the BACE group (13.4 months, 95% CI 11.6-15.2) ( P=0.001). The median PFS was also significantly longer in the BACE+A group (9.0 months, 95% CI 7.3-10.7) compared to the BACE group (6.1 months, 95% CI 4.9-7.3) ( P=0.001). At 6 and 12 months post-first BACE, the ORR (43.5%, 40.0%) and DCR (89.1%, 83.3%) were significantly higher in the BACE+A group than in the BACE group (ORR: 20.8%, 14.8%; DCR: 66.7%, 59.3%) (all P<0.05). Multivariate Cox regression identified treatment with BACE+A ( HR=0.42, 95% CI 0.27-0.72, P=0.002), tumor stage ( HR=1.80, 95% CI 1.05-3.07, P=0.031), presence of pre-existing complications requiring intervention ( HR=2.72, 95% CI 1.65-4.50, P<0.001), and >2 BACE procedures ( HR=0.32, 95% CI 0.15-0.68, P=0.003) as independent factors influencing OS. Treatment with BACE+A ( HR=0.49, 95% CI 0.32-0.76, P=0.001), tumor stage ( HR=1.72, 95% CI 1.07-2.77, P=0.025), multi-arterial tumor blood supply ( HR=2.76, 95% CI 1.76-4.31, P<0.001), and>2 BACE procedures ( HR=0.40, 95% CI 0.22-0.71, P=0.002) were independent factors influencing PFS. There was no significant difference in BACE-related adverse events between the two groups (all P>0.05). Hypertension, fatigue, hand-foot syndrome, and anorexia were common anlotinib-specific adverse reactions in the combination group, but no grade 4 or higher adverse reactions were observed. Conclusions:BACE combined with anlotinib demonstrates superior efficacy compared to BACE alone in treating advanced NSCLC, significantly prolonging OS and PFS. The safety profile is manageable, with adverse events remaining within tolerable limits.
4.Analysis of short-term efficacy of drug-coated balloon in the treatment of atherosclerotic renal artery stenosis
Haozhe ZHANG ; Qiongfang CHANG ; Juanfang LIU ; Xinwei HAN ; Jianhao ZHANG
Journal of Practical Radiology 2025;41(9):1541-1544
Objective To explore the feasibility,safety and efficacy of paclitaxel drug-coated balloon(DCB)in the treatment of atherosclerotic renal artery stenosis(ARAS).Methods A total of 28 patients with ARAS were selected.Balloon angioplasty was performed using a paclitaxel DCB at the site of renal artery stenosis(RAS)in these patients.Subsequently,a follow-up study was conducted to monitor various parameters of the patients,including vascular restenosis,blood pressure,the types of antihypertensive medications and renal function.The feasibility,efficacy and safety of balloon angioplasty using DCB in the treatment of ARAS were analyzed.Results Twenty-eight patients underwent 33 DCB balloon angioplasty.In one patient,there was no significant improve-ment in the degree of RAS,and thus further treatment with renal artery stenting was administered.The remaining patients all achieved both anatomical and hemodynamic success,with the degree of vascular stenosis at the lesion site decreasing from(79.74±5.13)%to(8.32±4.67)%,and the surgical success rate was 96.97%.The systolic/diastolic blood pressure of the patients was(179.16±30.65)mmHg/(108.26±20.93)mmHg(1 mmHg=0.133 kPa)at 24 hours postoperatively,(131.11±12.99)mmHg/(80.11±7.12)mmHg at 3 months postoperatively,(134.16±11.37)mmHg/(78.68±4.79)mmHg at 6 months postoperatively,and(133.37±12.71)mmHg/(80.11±4.84)mmHg at 12 months postoperatively.In comparison with the preoperative blood pressure of(184.63±27.64)mmHg/(109.11±22.26)mmHg,there was no significant decrease in blood pressure at 24 hours postoperatively,and the difference was not statistically significant.However,at 3,6,and 12 months postoperatively,the patients'blood pressure was signif-icantly lower than that before the operation,and all the differences were statistically significant.The glomerular filtration rate(GFR)was(36.19±18.32)mL/min at 24 hours postoperatively,(35.96±18.51)mL/min at 3 months postoperatively,(36.23±19.30)mL/min at 6 months postoperatively,and(35.59±18.26)mL//min at 12 months postoperatively,which all elevated compared with the preoperative GFR of(28.31±14.67)mL/min,and the differences were statistically significant(P<0.05).At 3,6,and 12 months postoperatively,the vascular patency rate was 100%as indicated by multifunctional color Doppler ultrasound examination or renal artery computed tomography angiography(CTA).No relevant com-plications and postoperative adverse events,such as renal artery rupture or dissection,renal artery thrombosis and acute renal insuffi-ciency,occurred in all 28 patients.Conclusion The paclitaxel DCB is safe and reliable for the treatment of ARAS and has remarkable curative effects,and it can be used as an effective vascular treatment regimen for ARAS.
5.Mechanism of Naringenin in amelioraing glucocorticoid-induced osteonecrosis of the femoral head by regulating HO-1/HIF-1α/VEGF axis
Xinwei ZHANG ; Ming SONG ; Hongxun ZHU ; Shouping DAI ; Yusong ZHANG ; Biaofang WEI
Immunological Journal 2025;41(11):769-779,792
Objective To investigate the mechanism of Naringenin(NGN)in the treatment of steroid(glucosteroid)-induced osteonecrosis of the femoral head(SONFH).Methods A SONFH rat model was established using Methylprednisolone(MPS)treatment,followed by intervention with NGN and zinc protoporphyrin(ZnPP).Micro-CT was used to analyze the morphological changes in femoral head tissues,and the levels of osteocalcin(OCN)in rat serum as well as heme oxygenase-1(HO-1)and hypoxia-inducible factor-1α(HIF-1α)in femoral bone tissue were measured.A cellular model was constructed by treating MC3T3-E1 cells with Dexamethasone(DEX),followed by NGN intervention.Bioinformatics analysis combined with molecular docking technology was used to predict the target of NGN,and the Pulldown experiment was performed for validation.The expression of HO-1 was knocked down through cell transfection,to analyze the viability,proliferation,apoptosis,and migration of MC3T3-E1 cells,and angiogenesis assays were conducted to evaluate the angiogenic potential of human umbilical vein endothelial cells(HUVECs).Results Micro-CT analysis revealed that,compared with the control group,the trabecular thickness and trabecular number were significantly reduced in the MPS group,while the bone surface area/bone volume ratio and trabecular separation were significantly increased(P<0.001).In vitro experimental results indicated that DEX inhibited the proliferation of MC3T3-E1 cells,promoted cell apoptosis,and increased reactive oxygen species generation(P<0.01),and that DEX suppressed the formation of mineralized nodules,a key indicator of osteogenic differentiation,and downregulated the expression of osteogenesis-related genes(Runt-related transcription factor 2,osteopontin,osteocalcin)(P<0.01).However,NGN treatment partially reversed these effects.DEX significantly inhibited the migration of HUVECs,angiogenesis,and the expression of angiogenesis-related markers(platelet endothelial cell adhesion molecule-1,vascular endothelial growth factor,and von Willebrand factor)(P<0.01).In contrast,NGN treatment did not significantly affect the aforementioned effects,but the treatment with NGN conditioned medium[CM(NGN)]partially reversed these effects(P<0.01).Bioinformatics analysis combined with Pulldown assay results indicated that HO-1 was the target of NGN.DEX treatment significantly downregulated the expression of HO-1,while NGN intervention partially counteracted the inhibitory effect induced by DEX(P<0.01);knockdown of HO-1 negated the therapeutic effects of NGN(P<0.01).Compared with MPS administration alone,the combined administration of NGN and MPS upregulated the expression of HO-1 and HIF-1α in rat femoral head tissues.However,the HO-1 inhibitor ZnPP further upregulated the expression of HO-1 but downregulated the protein level of hypoxia-inducible factor-α(HIF-1α)(P<0.01).Conclusion NGN exerts its therapeutic effects on SONFH by activating the expression and activity of HO-1,which regulates the HIF-1α/VEGF pathway to promote osteoblast differentiation,bone formation,and angiogenesis.
6.Mechanism of Naringenin in amelioraing glucocorticoid-induced osteonecrosis of the femoral head by regulating HO-1/HIF-1α/VEGF axis
Xinwei ZHANG ; Ming SONG ; Hongxun ZHU ; Shouping DAI ; Yusong ZHANG ; Biaofang WEI
Immunological Journal 2025;41(11):769-779,792
Objective To investigate the mechanism of Naringenin(NGN)in the treatment of steroid(glucosteroid)-induced osteonecrosis of the femoral head(SONFH).Methods A SONFH rat model was established using Methylprednisolone(MPS)treatment,followed by intervention with NGN and zinc protoporphyrin(ZnPP).Micro-CT was used to analyze the morphological changes in femoral head tissues,and the levels of osteocalcin(OCN)in rat serum as well as heme oxygenase-1(HO-1)and hypoxia-inducible factor-1α(HIF-1α)in femoral bone tissue were measured.A cellular model was constructed by treating MC3T3-E1 cells with Dexamethasone(DEX),followed by NGN intervention.Bioinformatics analysis combined with molecular docking technology was used to predict the target of NGN,and the Pulldown experiment was performed for validation.The expression of HO-1 was knocked down through cell transfection,to analyze the viability,proliferation,apoptosis,and migration of MC3T3-E1 cells,and angiogenesis assays were conducted to evaluate the angiogenic potential of human umbilical vein endothelial cells(HUVECs).Results Micro-CT analysis revealed that,compared with the control group,the trabecular thickness and trabecular number were significantly reduced in the MPS group,while the bone surface area/bone volume ratio and trabecular separation were significantly increased(P<0.001).In vitro experimental results indicated that DEX inhibited the proliferation of MC3T3-E1 cells,promoted cell apoptosis,and increased reactive oxygen species generation(P<0.01),and that DEX suppressed the formation of mineralized nodules,a key indicator of osteogenic differentiation,and downregulated the expression of osteogenesis-related genes(Runt-related transcription factor 2,osteopontin,osteocalcin)(P<0.01).However,NGN treatment partially reversed these effects.DEX significantly inhibited the migration of HUVECs,angiogenesis,and the expression of angiogenesis-related markers(platelet endothelial cell adhesion molecule-1,vascular endothelial growth factor,and von Willebrand factor)(P<0.01).In contrast,NGN treatment did not significantly affect the aforementioned effects,but the treatment with NGN conditioned medium[CM(NGN)]partially reversed these effects(P<0.01).Bioinformatics analysis combined with Pulldown assay results indicated that HO-1 was the target of NGN.DEX treatment significantly downregulated the expression of HO-1,while NGN intervention partially counteracted the inhibitory effect induced by DEX(P<0.01);knockdown of HO-1 negated the therapeutic effects of NGN(P<0.01).Compared with MPS administration alone,the combined administration of NGN and MPS upregulated the expression of HO-1 and HIF-1α in rat femoral head tissues.However,the HO-1 inhibitor ZnPP further upregulated the expression of HO-1 but downregulated the protein level of hypoxia-inducible factor-α(HIF-1α)(P<0.01).Conclusion NGN exerts its therapeutic effects on SONFH by activating the expression and activity of HO-1,which regulates the HIF-1α/VEGF pathway to promote osteoblast differentiation,bone formation,and angiogenesis.
7.Research progresses in coronary CT angiography of coronary atherosclerotic plaques
Mengyuan BAO ; Yongshun WU ; Xinwei ZHANG ; Linxin ZHANG ; Yan XING
Chinese Journal of Interventional Imaging and Therapy 2025;22(9):613-616
Coronary CT angiography(CCTA)plays an important role in study of coronary atherosclerotic plaques,which can be used to assess plaque composition and vulnerability,hence predicting major adverse cardiovascular events.Furthermore,CCTA combined with radiomics analysis is helpful for optimizing cardiovascular risk stratification.The research progresses in CCTA of coronary atherosclerotic plaques were reviewed in this article.
8.Diagnostic value of combined model based on clinicopathological and MRI features in BRCA-mutated ovarian cancer
Hao ZHANG ; Xiaohong CHEN ; Xinwei ZHONG ; Yi CHEN ; Bowen YUE ; Shourang CHEN ; Wenzhong HOU ; Zhiqi YANG ; Xiaofeng CHEN
Journal of Practical Radiology 2025;41(2):246-250
Objective To explore the diagnostic value of a combined model based on clinicopathological and MRI features in BRCA-mutated ovarian cancer.Methods The data of 132 patients with ovarian cancer who underwent pathology and BRCA gene tes-ting were analyzed retrospectively,including 52 cases of BRCA mutation group and 80 cases of BRCA wild group.The differences of MRI features and clinicopathological features between BRCA mutation group and BRCA wild group were compared.Binary logistic regression was used to construct a joint prediction model and analyze its diagnostic efficiency.Results There were significant differ-ences in cytokeratin 7(CK7),estrogen receptor(ER),Ki-67 and lymphovascular invasion(LVI)between the BRAC mutation group and the BRAC wild group(P<0.05).Univariate analysis showed that CK7,ER,Ki-67,LVI and the apparent diffusion coefficient(ADC)value of the cystic part of tumor were risk factors for BRCA-mutated ovarian cancer.The combined model based on CK7,ER,Ki-67,LVI,and the ADC value of the cystic part of tumor for the diagnosis of BRCA-mutated ovarian cancer had an area under the curve(AUC)of 0.765.Conclusion CK7,ER,Ki-67,LVI and the ADC value of the cystic part of tumor are risk factors for BRCA-mutated ovarian cancer.The combined model based on the above characteristics demonstrates good diagnostic efficacy for BRCA-mutated ovarian cancer.
9.A Case of Hypoparathyroidism With Hypocalcemic Heart Failure Caused by DiGeorge Syndrome
Xiru LIAN ; Liang ZHANG ; Chunfei ZHENG ; Wenping ZHAO ; Xinwei JIA ; Zhanqi WANG ; Xiangxin LI
Chinese Circulation Journal 2025;40(2):186-189
DiGeorge(DGS)syndrome is an autosomal dominant disorder caused by 22q11.2 microdeletions,most patients developed the disease in childhood.22q11.2 deletion syndrome,and the mutation types are dominated by haploid deletion of this gene.We report a young patient with hypoparathyroidism(parathyroidism)induced by DGS syndrome combined with hypocalcemic heart failure.Genetic testing revealed pathogenic copy number variants associated with the clinical phenotype of the subject.About 2 674 kb of deletion variation was detected at q11.21 position on chromosome 22,which contained the TBX1 gene and was a pathogenic mutation.This paper discusses the clinical features,pathogenesis and current treatment of DGS,and emphasizes the importance of early screening,early diagnosis and treatment,and regular follow-up of heart failure,aiming to enhance the awareness of clinicians and geneticists on DGS syndrome.
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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