1.Psychological stress-activated NR3C1/NUPR1 axis promotes ovarian tumor metastasis.
Bin LIU ; Wen-Zhe DENG ; Wen-Hua HU ; Rong-Xi LU ; Qing-Yu ZHANG ; Chen-Feng GAO ; Xiao-Jie HUANG ; Wei-Guo LIAO ; Jin GAO ; Yang LIU ; Hiroshi KURIHARA ; Yi-Fang LI ; Xu-Hui ZHANG ; Yan-Ping WU ; Lei LIANG ; Rong-Rong HE
Acta Pharmaceutica Sinica B 2025;15(6):3149-3162
Ovarian tumor (OT) is the most lethal form of gynecologic malignancy, with minimal improvements in patient outcomes over the past several decades. Metastasis is the leading cause of ovarian cancer-related deaths, yet the underlying mechanisms remain poorly understood. Psychological stress is known to activate the glucocorticoid receptor (NR3C1), a factor associated with poor prognosis in OT patients. However, the precise mechanisms linking NR3C1 signaling and metastasis have yet to be fully elucidated. In this study, we demonstrate that chronic restraint stress accelerates epithelial-mesenchymal transition (EMT) and metastasis in OT through an NR3C1-dependent mechanism involving nuclear protein 1 (NUPR1). Mechanistically, NR3C1 directly regulates the transcription of NUPR1, which in turn increases the expression of snail family transcriptional repressor 2 (SNAI2), a key driver of EMT. Clinically, elevated NR3C1 positively correlates with NUPR1 expression in OT patients, and both are positively associated with poorer prognosis. Overall, our study identified the NR3C1/NUPR1 axis as a critical regulatory pathway in psychological stress-induced OT metastasis, suggesting a potential therapeutic target for intervention in OT metastasis.
2.Diagnosis of coronary artery lesions in children based on Z-score regression model.
Yong WANG ; Jia-Ying JIANG ; Yan DENG ; Bo LI ; Ping SHUAI ; Xiao-Ping HU ; Yin-Yan ZHANG ; Han WU ; Lu-Wei YE ; Qian PENG
Chinese Journal of Contemporary Pediatrics 2025;27(2):176-183
OBJECTIVES:
To construct a Z-score regression model for coronary artery diameter based on echocardiographic data from children in Sichuan Province and to establish a Z-score calculation formula.
METHODS:
A total of 744 healthy children who underwent physical examinations at Sichuan Provincial People's Hospital from January 2020 to December 2022 were selected as the modeling group, while 251 children diagnosed with Kawasaki disease at the same hospital from January 2018 to December 2022 were selected as the validation group. Pearson correlation analysis was conducted to analyze the relationships between coronary artery diameter values and age, height, weight, and body surface area. A regression model was constructed using function transformation to identify the optimal regression model and establish the Z-score calculation formula, which was then validated.
RESULTS:
The Pearson correlation analysis showed that the correlation coefficients for the diameters of the left main coronary artery, left anterior descending artery, left circumflex artery, and right coronary artery with body surface area were 0.815, 0.793, 0.704, and 0.802, respectively (P<0.05). Among the constructed regression models, the power function regression model demonstrated the best performance and was therefore chosen as the optimal model for establishing the Z-score calculation formula. Based on this Z-score calculation formula, the detection rate of coronary artery lesions was found to be 21.5% (54/251), which was higher than the detection rate based on absolute values of coronary artery diameter. Notably, in the left anterior descending and left circumflex arteries, the detection rate of coronary artery lesions using this Z-score calculation formula was higher than that of previous classic Z-score calculation formulas.
CONCLUSIONS
The Z-score calculation formula established based on the power function regression model has a higher detection rate for coronary artery lesions, providing a strong reference for clinicians, particularly in assessing coronary artery lesions in children with Kawasaki disease.
Humans
;
Male
;
Female
;
Child, Preschool
;
Child
;
Coronary Artery Disease/diagnostic imaging*
;
Infant
;
Mucocutaneous Lymph Node Syndrome
;
Regression Analysis
;
Coronary Vessels/diagnostic imaging*
;
Echocardiography
;
Adolescent
3.Comprehensive Analysis of Oncogenic, Prognostic, and Immunological Roles of FANCD2 in Hepatocellular Carcinoma: A Potential Predictor for Survival and Immunotherapy.
Meng Jiao XU ; Wen DENG ; Ting Ting JIANG ; Shi Yu WANG ; Ru Yu LIU ; Min CHANG ; Shu Ling WU ; Ge SHEN ; Xiao Xue CHEN ; Yuan Jiao GAO ; Hongxiao HAO ; Lei Ping HU ; Lu ZHANG ; Yao LU ; Wei YI ; Yao XIE ; Ming Hui LI
Biomedical and Environmental Sciences 2025;38(3):313-327
OBJECTIVE:
Hepatocellular carcinoma (HCC) is sensitive to ferroptosis, a new form of programmed cell death that occurs in most tumor types. However, the mechanism through which ferroptosis modulates HCC remains unclear. This study aimed to investigate the oncogenic role and prognostic value of FANCD2 and provide novel insights into the prognostic assessment and prediction of immunotherapy.
METHODS:
Using clinicopathological parameters and bioinformatic techniques, we comprehensively examined the expression of FANCD2 macroscopically and microcosmically. We conducted univariate and multivariate Cox regression analyses to identify the prognostic value of FANCD2 in HCC and elucidated the detailed molecular mechanisms underlying the involvement of FANCD2 in oncogenesis by promoting iron-related death.
RESULTS:
FANCD2 was significantly upregulated in digestive system cancers with abundant immune infiltration. As an independent risk factor for HCC, a high FANCD2 expression level was associated with poor clinical outcomes and response to immune checkpoint blockade. Gene set enrichment analysis revealed that FANCD2 was mainly involved in the cell cycle and CYP450 metabolism.
CONCLUSION
To the best of our knowledge, this is the first study to comprehensively elucidate the oncogenic role of FANCD2. FANCD2 has a tumor-promoting aspect in the digestive system and acts as an independent risk factor in HCC; hence, it has recognized value for predicting tumor aggressiveness and prognosis and may be a potential biomarker for poor responsiveness to immunotherapy.
Humans
;
Carcinoma, Hepatocellular/diagnosis*
;
Liver Neoplasms/diagnosis*
;
Immunotherapy
;
Fanconi Anemia Complementation Group D2 Protein/metabolism*
;
Prognosis
;
Male
;
Female
;
Middle Aged
;
Biomarkers, Tumor/metabolism*
4.Single-stage treatment of upper limb lymphedema following breast cancer surgery using superficial circumflex iliac artery perforator-based vascularized lymph node transfer combined with lymphaticovenular anastomosis and liposuction.
Zongcan CHEN ; Junzhe CHEN ; Yuanyuan WANG ; Lingli JIANG ; Xiangkui WU ; Hai LI ; Shune XIAO ; Chengliang DENG
Chinese Journal of Reparative and Reconstructive Surgery 2025;39(9):1114-1121
OBJECTIVE:
To compare the effectiveness of single-stage vascularized lymph node transfer (VLNT) combined with lymphaticovenular anastomosis (LVA) and liposuction (LS) (3L) versus LVA combined with LS (2L) for the treatment of moderate-to-late stage upper limb lymphedema following breast cancer surgery.
METHODS:
A retrospective analysis was conducted on the clinical data of 16 patients with moderate-to-late stage upper limb lymphedema after breast cancer surgery, treated between June 2022 and June 2024, who met the selection criteria. Patients were divided into 3L group (n=7) and 2L group (n=9) based on the surgical approach. There was no significant difference (P>0.05) in baseline data between the groups, including age, body mass index, duration of edema, volume of liposuction, International Society of Lymphology (ISL) stage, preoperative affected limb volume, preoperative circumferences of the affected limb at 12 levels (from 4 cm distal to the wrist to 42 cm proximal to the wrist), preoperative Lymphoedema Quality of Life (LYMQoL) score, and frequency of cellulitis episodes. The 2L group underwent LS on the upper arm and proximal forearm and LVA on the middle and distal forearm. The 3L group received additional VLNT in the axilla, with the groin serving as the donor site. Outcomes were assessed included the change in affected limb volume at 12 months postoperatively, and comparisons of limb circumferences, LYMQoL score, and frequency of cellulitis episodes between preoperative and 12-month postoperative. Ultrasound evaluation was performed at 12 months in the 3L group to assess lymph node viability.
RESULTS:
Both groups were followed up 12-20 months, with an average of 15.13 months. There was no significant difference in the follow-up time between the groups (t=-1.115, P=0.284). All surgical incisions healed by first intention. No adverse events, such as flap infection or necrosis, occurred in the 3L group. At 12 months after operation, ultrasound confirmed good viability of the transferred lymph nodes in the 3L group. Palpation revealed significant improvement in skin fibrosis and improved skin softness in both groups. Affected limb volume significantly decreased in both groups postoperatively (P<0.05). The reduction in limb volume significantly greater in the 3L group compared to the 2L group (P<0.05). Circumferences at all 12 measured levels significantly decreased in both groups compared to preoperative values (P<0.05). The reduction in circumference at all 12 levels was better in the 3L group than in the 2L group, with significant differences observed at 7 levels (8, 12, 16, 30, 34, 38, and 42 cm) proximal to the wrist (P<0.05). Both groups showed significant improvement in the frequency of cellulitis episodes and LYMQoL scores postoperatively (P<0.05). While the improvement in LYMQoL scores at 12 months did not differ significantly between groups (P>0.05), the reduction in cellulitis episodes was significantly greater in the 3L group compared to the 2L group (P<0.05).
CONCLUSION
The combination of VLNT+LVA+LS provides more durable and comprehensive outcomes for moderate-to-late stage upper limb lymphedema after breast cancer surgery compared to LVA+LS, offering an improved therapeutic solution for patients.
Humans
;
Female
;
Lipectomy/methods*
;
Retrospective Studies
;
Anastomosis, Surgical/methods*
;
Lymphedema/etiology*
;
Middle Aged
;
Upper Extremity/surgery*
;
Breast Neoplasms/surgery*
;
Lymph Nodes/blood supply*
;
Adult
;
Lymphatic Vessels/surgery*
;
Iliac Artery/surgery*
;
Postoperative Complications/surgery*
;
Perforator Flap/blood supply*
;
Treatment Outcome
;
Mastectomy/adverse effects*
;
Quality of Life
;
Aged
5.Chimeric flap with a perforator of superficial circumflex iliac artery in treatment of complex tissue defects in Achilles tendon region: 9 cases report
Hai LI ; Zairong WEI ; Shun'e XIAO ; Xiangkui WU ; Bihua WU ; Chengliang DENG
Chinese Journal of Microsurgery 2025;48(1):25-30
Objective:To investigate the clinical effect of chimeric flap with perforating branch of superficial circumflex iliac artery in reconstruction of complex tissue defect in Achilles tendon region.Methods:A retrospective analysis was conducted on 9 patients who underwent reconstruction of composite tissue defects in Achilles tendon region with the perforator flap of lateral femoral circumflex artery, from January 2017 to January 2023 in the Department of Burns and Plastic Surgery, Affiliated Hospital of Zunyi Medical University. The patients were 7 males and 2 females and at 16 to 54 years, with 36.6 years in average. Causes of injury were: 5 patients by traffic accident, 2 by machine strangulation and 1 by heavy object, also 1 defect was caused by infection. The soft tissue defects were 4.5 cm×6.0 cm to 8.0 cm×12.0 cm in size, and the defects of Achilles tendon were 4.5 cm to 11.5 cm, with an average of 6.8 cm in length. The wounds were managed with VSD after primary surgery, and then stage Ⅰ surgery were followed to reconstruct the defects in Achilles tendon region with perforator flap of lateral femoral circumflex artery. The sizes of chimeric flaps were 6.5 cm×5.0 cm to 18.0 cm×6.5 cm, including 3 lobuated flaps. The fascia grafts were 5.5 cm to 12.0 cm, with an average of 7.0 cm. Scheduled postoperative follow-ups were conducted in outpatient clinic and via telephone or WeChat interviews, together with the advices for rehabilitation training. Postoperative functions of the Achilles tendon and ankle function were evaluated using Arner-Lindholm criteria and the ankle and hindfoot functional score of American Orthopaedic Foot and Ankle Society (AOFAS).Results:All of the 9 chimeric flaps survived and all the donor sites had primary healing. All patients were entered into the postoperative follow-up for 10 to 66 months, with 14.6 months in average. One chimeric flap was found bulky and a flap thinning surgery was further carried out in 3 months under the request of patient. The rest of patients were satisfied with the appearance and texture of the flaps. The range of ankle plantar flexions was found at 42.9°±1.8° and the dorsal flexions was at 16.2°±2.5°. There was no significant decrease in circumference and no discomfort or muscle re-rupture in the affected legs. Thompson test was found negative. There was no wound related complications during follow-up. Reconstructive surgery were found effective based on Arner-Lindholm criteria, with excellent in 6 patients, good in 2 and 1 in fair. The ankle and subtalar joint function was evaluated according to the ankle-hindfoot function score of AOFAS, with 7 patients scored 90-100 point and 2 scored 75-89 point, and 7 patients were in excellent and 2 in good.Conclusion:The chimeric perforator flap of lateral femoral circumflex artery is safe and reliable in reconstruction of composite tissue defects in Achilles tendon region. It can effectively restore the function and appearance of ankle and hindfoot in patients with composite tissue defects in the Achilles tendon region.
6.Thin perforator flap of superficial circumflex iliac artery with venous superdrainage in reconstruction of the soft tissue defect in extremities: a report of 20 cases
Hai LI ; Cheng ZHANG ; Chengliang DENG ; Shun'e XIAO ; Xiangkui WU ; Lingli JIANG ; Zairong WEI
Chinese Journal of Microsurgery 2025;48(4):382-387
Objective:To explore the clinical effect of thin superficial circumflex iliac artery perforator flap with venous superdrainage in treatment of wound in extremities.Methods:Clinical data of 20 patients who were treated from January 2018 to January 2024 in the Department of Burns and Plastic Surgery, Affiliated Hospital of Zunyi Medical University were retrospectively analysed. The soft tissue defects in extremity were reconstructed with thin perforator flaps of superficial iliac circumflex artery with venous superdrainage. There were 13 patients with upper limb defects and 7 with ankle defects. Of the defects, 12 were of trauma, 2 of tumour resection, 3 of scar release surgery and 3 of donor flat harvesting. The defects ranged from 5.0 cm×4.0 cm to 18.0 cm×7.0 cm in size. During the surgery, portable Doppler was used to detect the perforating branches of the superficial rotational iliac artery and design flaps. The flaps were 5.0 cm×5.0 cm to 20.0 cm×7.0 cm in size, including 16 single-lobed flaps, 3 double-lobed flaps and 1 triple-lobed flaps. The flaps were 2-7 mm in thickness, with an average of 4 mm. Based on the distribution of perforators, the flaps carried the superficial helioiliac artery and the accompanying vein, as well as the superficial helioiliac vein. The superficial helioiliac artery and the accompanying vein were anastomosed during the surgery, and the superficial helioiliac vein was anastomosed to the superficial or deep vein of the recipient site for superreflux. The donor sites were all directly sutured and closed. Postoperative follow-up was conducted by visits of outpatient clinic, and via telephone and WeChat interviews. The survival and appearance of the flaps and complications were observed.Results:A total of 22 arteries and 42 veins of the 20 flaps were anastomosed in surgery. All flaps survived. The donor sites were closed in the first stage. No vascular compromise occurred. One patient had early exudate under the flap on the exposed wound of interphalangeal joint, which healed after routine dressing change and drainage. All patients were included in the postoperative follow-up, with a peroid over 5 to 40 months, at 10.8 months in average. The flaps were thin and soft, with good wear resistance and without pigmentation. The healing of donor sites was good except 1 donore site that had early lymphatic leakage, which was cured after compression and drainage. A linear scar left at the donor sites and it was acceptable to the patients.Conclusion:The treatment of defective soft tissue wounds in extremities using thin perforator flap with venous superdrainage of the superficial circumflex iliac artery is safe and feasible. No further flap thinning surgery is required, and there is a reliable clinical effect.
7.Clinical efficacy of modified superior gluteal artery perforator flap with V-Y advancement for the reconstruction of sacrococcygeal pilonidal sinus
Hai LI ; Xiangkui WU ; Chengliang DENG ; Shun’e XIAO ; Zairong WEI
Chinese Journal of Plastic Surgery 2025;41(11):1175-1182
Objective:To investigate the clinical efficacy of the modified superior gluteal artery perforator (SGAP) flap V-Y advancement technique in the repair of sacrococcygeal pilonidal sinus.Methods:A retrospective analysis was conducted on the clinical data of patients who underwent modified SGAP flap V-Y advancement for the reconstruction of sacrococcygeal pilonidal sinus wounds at the Department of Burns and Plastic Surgery, Affiliated Hospital of Zunyi Medical University, between January 2019 and January 2025. The surgical procedure involved two stages. During the first stage, the sinus tract was preoperatively marked with methylene blue, followed by complete excision of the sinus and surrounding involved tissues. The resultant wound was then treated with vacuum sealing drainage for 3 to 5 days. The second stage consisted of flap repair. A portable Doppler ultrasound was used to identify and mark the superior gluteal artery perforator on one side of the wound. A V-shaped flap was designed transversely, with the base corresponding to the length of the subcutaneous tissue defect on the wound side and the superior and inferior borders extending laterally along the wound margins, ensuring the 1-2 SGAP was included within the flap. Intraoperatively, the wound was re-debrided. The epidermis was removed from the base of the V-shaped flap, and the dermis and subcutaneous tissue were dissected. The dermis and superficial subcutaneous tissue of the contralateral wound edge were also dissected. The flap was then advanced in a V-Y fashion to cover the wound. The de-epithelialized portion of the flap was imbricated and sutured to the contralateral skin edge near the midline. The donor site at the apex of the V-shaped flap was closed directly after subcutaneous tissue tension relief, resulting in a Y-shaped closure. Postoperative flap survival, incision healing, and complications were observed. At the final follow-up, flap appearance, texture, surgical site scarring, and sinus recurrence were assessed, and patient satisfaction with the surgical outcome was investigated.Results:A total of eight patients were included in the study, comprising six males and two females, with ages ranging from 14 to 41 years (mean 22.4 years). Two patients presented with multiple sinus tracts, two with chronic sinus tracts and recurrent inflammation with scar formation, and four with postoperative recurrence. The area of the wound defect after debridement ranged from 4.5 cm×5.0 cm to 6.5 cm×8.0 cm. The flap dimensions ranged from 4.5 cm × 6.0 cm to 6.5 cm × 12.5 cm, with a mean V-shaped flap apex angle of 35° ± 2° (range 20° to 60°). In one patient, slight exudate was observed at the base of the flap, which healed after 12 days of dressing changes and drainage. The remaining flaps survived successfully, and primary closure was achieved in both the donor and recipient sites. The patients were followed for a period of 3 to 64 months, with a mean follow-up duration of 10.7 months. In all patients, the sacrococcygeal flap showed no evidence of bulkiness, exhibiting a color similar to the surrounding skin and a soft texture. Linear scars remained at the surgical site, with no complaints of itching, pain, or other discomfort affecting daily life. No recurrence of pilonidal sinus was observed during the follow-up period. Patients reported satisfaction with the appearance of their buttocks.Conclusion:The modified SGAP flap V-Y advancement technique for the repair of sacrococcygeal pilonidal sinus effectively elevates the gluteal cleft, facilitates off-midline closure, demonstrates reliable result, and provides a high level of patient satisfaction.
8.Repair effect of oleuropein on OGD/R-induced neuronal damage by regulating Hippo-YAP signaling pathway
Yanling DENG ; Zhigang WU ; Qian XIAO ; Ke ZHANG
Chinese Journal of Immunology 2025;41(11):2607-2612,2618
Objective:To investigate the effects of oleuropein(OE)on oxygen glucose deprivation/reperfusion(OGD/R)in-duced neuronal damage and Hippo-Yes associated protein(YAP)signaling pathway.Methods:Neurons were grouped into Control group,OGD/R group,OE-L group,OE-M group,OE-H group and OE-H+YAP inhibitor(VP)group;detection of cell activity;cell proliferation and apoptosis were detected in each group;the levels of inflammatory factors and oxidative stress were detected;the ex-pression levels of Cyclin D1,cell cycle negative regulator(P21),B cell lymphoma 2(Bcl-2),Bcl-2 associated X protein(Bax),YAP and transcription coactivator with PDZ-binding motif(TAZ)were detected.Results:In OGD/R group,deep staining of neuronal cell nuclei,EdU positive rate,Cyclin D1,Bcl-2,YAP and TAZ expressions were decreased,P21,TNF-α,IL-6,MDA,SOD,apop-tosis rate and Bax expression were increased(P<0.05).The cell nucleus was gradually restored to normal by OE,the blue staining was gradually lightest,the EdU positive rate,Cyclin D1,Bcl-2,YAP and TAZ levels were increased,and the expressions of P21,TNF-α,IL-6,MDA,SOD,apoptosis rate and Bax were decreased(P<0.05);VP reverse protective effect of OE on OGD/R-induced neuronal injury.Conclusion:OE can improve OGD/R-induced neuronal damage by activating Hippo/YAP signaling pathway.
9.Prognostic value of single PET-CT after chemotherapy combined with immunotherapy in patients with non-small cell lung cancer treated with radiotherapy
Zhenghui MA ; Yuqi WU ; Guangqian JI ; Zongmei ZHOU ; Qinfu FENG ; Zefen XIAO ; Jima LYU ; Xin WANG ; Jianyang WANG ; Wenyang LIU ; Lei DENG ; Wenqing WANG ; Nan BI ; Junlin YI ; Tao ZHANG
Chinese Journal of Radiation Oncology 2025;34(11):1111-1116
Objective:To evaluate the role of a single PET-CT scan in predicting survival and prognosis in patients with non-small cell lung cancer (NSCLC) who did not undergo surgery but received radiotherapy after neoadjuvant chemotherapy combined with immunotherapy.Methods:A retrospective analysis was conducted on the data of 23 NSCLC patients treated at the Cancer Hospital of the Chinese Academy of Medical Sciences from May 2022 to June 2024. All patients were pathologically confirmed, received neoadjuvant chemotherapy combined with immunotherapy, did not undergo surgery for various reasons, and instead received radiotherapy. Each patient underwent only one PET-CT scan after neoadjuvant chemotherapy combined with immunotherapy and before radiotherapy. According to the maximum standardized uptake value (SUV max) on PET-CT, patients were divided into the low-uptake group (SUV max < 8, n=12) and high-uptake group (SUV max ≥ 8, n=11). Survival analysis was performed using the Kaplan-Meier method with survival curves plotted. Univariate analysis of influencing factors of survival was conducted using the Cox proportional hazards regression model. Clinical characteristics and survival outcomes of the two groups were compared, including progression-free survival (PFS) and overall survival (OS). Results:The 1-year PFS rates were 100% in the low-uptake group, 54.5% in the high-uptake group. This difference was statistically significant ( P=0.007). The 1-year and 2-year OS rates were both 100% in the low-uptake group, the 1-year and 2-year OS rates were both 90.9% in the high-uptake group, with no statistically significant difference ( P=0.394). Univariate Cox analysis identified age as an independent factor affecting PFS. Conclusions:For NSCLC patients who did not undergo surgical resection but received radiotherapy after neoadjuvant chemotherapy combined with immunotherapy, a single PET-CT scan before radiotherapy has potential value in predicting PFS. However, clinical studies with larger sample size and longer follow-up are required to evaluate its predictive value for OS.
10.Comparison of the toxicity and safety of protein derivatives from novel fusion strains of Mycobacterium tuberculosis
Hao-qi XU ; Jiang-tao DONG ; Jie ZHANG ; Fang WU ; Su LIANG ; Xiao-ling LIU ; Lan-ru GAO ; Ju WANG ; Hui ZHANG ; Jiang-dong WU ; Le ZHANG ; Xi-ling DENG ; Wan-jiang ZHANG
Chinese Journal of Zoonoses 2025;41(4):376-384
The objective of this study was to evaluate the toxicity and safety of novel Mycobacterium tuberculosis fusion strain protein derivatives,referred to as B/R strain active proteins.In cellular experiments,RAW264.7 cells were treated with each vaccine preparation,and apoptosis rates were measured.In subsequent animal experiments,C57BL/6 mice were immunized via subcutaneous injection,and their survival and body weight changes were monitored and recorded at 2,4,8,12,and 16 weeks.The lungs and spleens were harvested to calculate organ coefficients,and pathological examinations were conducted.At the eighth week of immunization,the mice were infected with high concentrations of BCG,and pathological changes in the lungs and spleens were observed 4 weeks post-infection.The apoptosis rate at 6 hours was significantly higher in the experimental group than the PBS group(P<0.05).At 12 and 24 hours,the apoptosis rate in the experimental group remained higher than that in the PBS group,although this difference was not statistically significant.After immunization,mice in all four groups exhibited normal growth patterns,as indicated by stable body weight changes.At 4 and 12 weeks post-immunization,the lung coefficients in the protein group were significantly higher than those in the PBS group at the same time points.Additionally,the lung coefficients in the BCG group were significantly elevated across all time periods(P<0.05).The spleen coefficients in the protein and BCG groups were significantly higher than those in the PBS group at 2,4,8,12,and 16 weeks,whereas the ICD B/R group showed higher spleen coefficients than the PBS group only at week 8(P<0.05).Pathological examination revealed normal lung and spleen tissues in the PBS group.However,during the 2-8 weeks immunization period,lung and spleen tissues in all experimental groups exhibited varying degrees of damage,which gradually diminished by 12-16 weeks.Notably,no tuberculosis nodules were observed in any experimental group.After infection with high concentrations of BCG,no overt pathological changes were observed on the surfaces of the lungs and spleens in any group.Microscopic examination revealed less severe pathological changes in the lungs and spleens of mice in the experimental groups than the PBS group.Furthermore,no statistically significant differences were observed between the protein group and the BCG group.Our findings suggested that the B/R strain active proteins'toxicity and safety profiles were comparable to those of BCG,and showed immunoprotective effects.This study provides an experimental foundation for the development of a novel tuberculosis vaccine.

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