1.Guideline for the diagnosis and treatment of vertebral refracture after percutaneous vertebral augmentation in elderly patients with osteoporotic thoracolumbar compression fractures (version 2025)
Yong YANG ; Xiaoguang ZHOU ; Qixin CHEN ; Jian CHEN ; Jian DONG ; Liangjie DU ; Shunwu FAN ; Jin FAN ; Zhong FANG ; Haoyu FENG ; Shiqing FENG ; Haishan GUAN ; Aiguo GAO ; Yanzheng GAO ; Yong HAI ; Da HE ; Dengwei HE ; Haiyi HE ; Dianming JIANG ; Xuewen KANG ; Bin LIN ; Baoge LIU ; Changqing LI ; Fang LI ; Li LI ; Fangcai LI ; Weishi LI ; Xiaoguang LIU ; Hongjian LIU ; Xinyu LIU ; Yong LIU ; Zhongjun LIU ; Shibao LU ; Xuhua LU ; Fei LUO ; Yuhai MA ; Keya MAO ; Xuexiao MA ; Bin MENG ; Xu NING ; Limin RONG ; Hongxun SANG ; Jun SHU ; Tiansheng SUN ; Dasheng TIAN ; Zheng WANG ; Bing WANG ; Linfeng WANG ; Qingde WANG ; Qinghe WANG ; Lan WEI ; Jigong WU ; Baoshan XU ; Youjia XU ; Guoyong YIN ; Jinglong YAN ; Feng YAN ; Cao YANG ; Huilin YANG ; Qiang YANG ; Bin ZHAO ; Jie ZHAO ; Yue ZHU ; Jianguo ZHANG ; Wenzhi ZHANG ; Zhongmin ZHANG ; Zhaomin ZHENG ; Yan ZENG ; Baorong HE ; Wei MEI
Chinese Journal of Trauma 2025;41(7):613-626
Vertebral refracture following percutaneous vertebral augmentation (PVA) is commonly seen in elderly patients with osteoporotic thoracolumbar compression fractures (OTLCF). It can lead to recurrent pain, loss of vertebral height, progression of kyphosis, and even neurological dysfunction, significantly impairing patients′ quality of life. Current diagnosis and treatment face multiple challenges, including high misdiagnosis rate, difficulty in choosing between surgical and non-surgical treatment options, lack of standardized surgical protocols, interference from intralesional bone cement during procedures, inadequate stability of internal fixation in osteoporotic bone, and suboptimal compliance of anti-osteoporotic therapy. Establishing a standardized diagnostic and therapeutic framework is urgently needed. To standardize the management process and improve outcomes for vertebral refractures after PVA in elderly OTLCF patients, Spinal Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized experts in the field to develop Guideline for the diagnosis and treatment of vertebral refracture after percutaneous vertebral augmentation in elderly patients with osteoporotic thoracolumbar compression fractures ( version 2025), based on current literature and clinical experience, and adhering to principles of scientific rigor and clinical applicability. A total of 11 recommendations were proposed, encompassing diagnosis, treatment, and rehabilitation of vertebral refracture after PVA in elderly patients with OTLCF, aiming to provide a foundation for a standardized management.
2.Construction and performance evaluation of a predictive model for post-traumatic hydrocephalus in patients with severe traumatic brain injury
Bin XU ; Xin WANG ; Jiahao LIAO ; Yuhai WANG ; Jinxu ZHOU
Chinese Journal of Trauma 2025;41(11):1059-1069
Objective:To develop a predictive model for the risk of post-traumatic hydrocephalus (PTH) in patients with severe traumatic brain injury (sTBI) and validate its predictive performance.Methods:A retrospective case control study was conducted to analyze the clinical data of 580 sTBI patients admitted to the 904th Hospital of the Joint Logistics Support Force of the PLA between January 2016 and December 2023, including 413 males and 167 females, aged 18-88 years [(54.3±14.6)years]. Patients were stratified into PTH group ( n=195) and non-PTH group ( n=385), based on the presence of PTH within 6 months after injury. Data collected from the two groups such as general baseline indicators, TBI-related clinical indicators (including surgical data), laboratory findings, and radiological features. Except for the data collected during the operation, all the above data are the results of the first examination at admission. Univariate analysis and Lasso regression analysis were used to screen predictors for the risk of PTH in sTBI patients. Subsequent multivariate Logistic regression was employed to identify predictors and construct a regression equation. Based on this equation, a nomogram prediction model was developed using the R language. Model discrimination was estimated through the receiver operating characteristic (ROC) curve, and calibration performance via the Hosmer-Lemeshow (H-L) goodness-of-fit test and calibration curve. Moreover, decision curve analysis (DCA) and clinical impact curve (CIC) were used for evaluating the clinical utility of the model. Results:Univariate analysis revealed statistically significant differences in 37 variables between the two groups, including age, age group, heart rate, oxygen saturation, Glasgow coma scale (GCS) score, left pupil size, right pupil size, pupillary light reflex, intracranial pressure (ICP) monitoring, type of decompressive craniectomy, neutrophil count, lymphocyte count, monocyte count, red blood cell count, platelet count, neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), neutrophil-lymphocyte-platelet ratio (N/LP), lymphocyte-to-monocyte ratio (LMR), systemic inflammation response index (SIRI), systemic immune-inflammation index (SII), fibrinogen (FIB), D-dimer, D-dimer-to-fibrinogen ratio (DFR), serum albumin, prognostic nutritional index, blood glucose, status of basal cisterns, midline shift, degree of midline shift, cerebral herniation, epidural hematoma (EDH), subdural hematoma (SDH), intraventricular hemorrhage (IVH), subarachnoid hemorrhage (SAH), modified Fisher grade, and skull fracture ( P<0.05). Lasso regression analysis identified 24 potential predictors for PTH, including age, GCS score, pupillary light reflex, type of decompressive craniectomy, monocyte count, platelet count, NLR, PLR, N/LP, LMR, SII, D-dimer, DFR, serum albumin, prognostic nutritional index, blood glucose, status of basal cisterns, degree of midline shift, cerebral herniation, EDH, SDH, IVH, modified Fisher grade and skull fracture. Multivariate Logistic regression analysis demonstrated that age, unilateral pupillary light reflex, absent pupillary light reflex, bilateral decompressive craniectomy, monocyte count, PLR, cerebral herniation, SDH, IVH, linear skull fracture and depressed skull fracture were independent risk factors for PTH. In contrast, serum albumin was identified as an independent protective factor for PTH ( P<0.05). The regression equation derived from these factors was: Logit[ P/(1- P)]=0.05×"age"+1.65×"unilateral pupillary light reflex"+2.79×"absent pupillary light reflex"+1.60×"bilateral decompressive craniectomy"+1.90×"monocyte count"+0.02×"PLR"-0.12×"serum albumin"+2.07×"cerebral herniation"+2.59×"SDH"+2.23×"IVH"+1.24×"linear skull fracture"+ 1.66×"depressed skull fracture"-22.61. The prediction model built upon this equation achieved an area under the ROC curve (AUC) of 0.95(95% CI 0.93, 0.97), with a sensitivity of 91.79%, specificity of 85.97%, and Youden′s index of 0.78. The H-L goodness-of-fit test indicated good calibration ( χ2=7.90, P=0.545). DCA results showed that the bias-corrected curve closely aligned with the actual curve and approximated the ideal curve, indicating a high clinical net benefit. Furthermore, CIC results demonstrated that with threshold probabilities greater than 60%, the number of patients identified as high-risk by the model highly corresponded with the actual number of patients who developed PTH. Conclusion:The prediction model incorporating age, unilateral pupillary light reflex, absent pupillary light reflex, bilateral decompressive craniectomy, monocyte count, PLR, serum albumin, cerebral herniation, SDH, IVH, linear skull fracture and depressed skull fracture exhibits robust predictive performance for PTH in sTBI patients.
3.Association between dietary diversity and cognitive function among the elderly based on group-based trajectory model
DONG Chunling ; WANG Chong ; GUAN Tianyuan ; LIU Yue ; LI Xueying ; ZHANG Yuhai
Journal of Preventive Medicine 2025;37(9):865-869
Objective:
To analyze the association between dietary diversity and cognitive function among the elderly using group-based trajectory model, so as to provide the basis for formulating dietary intervention strategies to prevent cognitive impairment.
Methods:
Based on the Chinese Longitudinal Healthy Longevity Survey (CLHLS) project, demographic information, lifestyle behaviors, psychological well-being, and activities of daily living of elderly individuals aged ≥65 years from 2008 to 2018 were collected. Dietary diversity was assessed using a food frequency questionnaire, with a score of <7 points defined as low dietary diversity. Cognitive function was evaluated using the Chinese version of the Mini-Mental State Examination (MMSE). A group-based trajectory model was established based on cognitive function scores from 2008 to 2018 to analyze the trajectories of cognitive function change. The association between dietary diversity and cognitive function was analyzed using a multinomial logistic regression model.
Results:
A total of 1 613 individuals were collected, with a median age was 72.00 (interquartile range, 10.00) years. There were 810 males (50.22%) and 803 females (49.78%). The group-based trajectory model analysis categorized the participants into three groups: the low-level normal group, the high-level normal group, and the slow-then-rapid decline group, comprising 796 (49.35%), 585 (36.27%), and 232 (14.38%) individuals, respectively. Among these groups, the numbers of individuals with low dietary diversity were 497 (62.44%), 311 (53.16%), and 166 (71.55%), respectively, with a statistically significant difference (P<0.05). Multinomial logistic regression analysis showed that after adjusting for demographic information, lifestyle behaviors, psychological well-being, and activities of daily living, compared with the high-level normal group, low dietary diversity was statistically associated with cognitive function in the slow-then-rapid decline group (OR=1.622, 95%CI: 1.103-2.384).
Conclusion
Low dietary diversity may increase the risk of cognitive impairment among the elderly.
4.Image-aware generative medical visual question answering based on image caption prompts.
Rui WANG ; Jiana MENG ; Yuhai YU ; Siwei HAN ; Xinghao LI
Journal of Biomedical Engineering 2025;42(3):560-566
Medical visual question answering (MVQA) plays a crucial role in the fields of computer-aided diagnosis and telemedicine. Due to the limited size and uneven annotation quality of the MVQA datasets, most existing methods rely on additional datasets for pre-training and use discriminant formulas to predict answers from a predefined set of labels. This approach makes the model prone to overfitting in low resource domains. To cope with the above problems, we propose an image-aware generative MVQA method based on image caption prompts. Firstly, we combine a dual visual feature extractor with a progressive bilinear attention interaction module to extract multi-level image features. Secondly, we propose an image caption prompt method to guide the model to better understand the image information. Finally, the image-aware generative model is used to generate answers. Experimental results show that our proposed method outperforms existing models on the MVQA task, realizing efficient visual feature extraction, as well as flexible and accurate answer outputs with small computational costs in low-resource domains. It is of great significance for achieving personalized precision medicine, reducing medical burden, and improving medical diagnosis efficiency.
Humans
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Image Processing, Computer-Assisted/methods*
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Diagnosis, Computer-Assisted/methods*
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Algorithms
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Telemedicine
5.Experimental study on the treatment of postmenopausal osteoporosis with low-frequency pulsed electromagnetic fields.
Zidong AN ; Liqiang WANG ; Yi WU ; Yongjie PANG ; Keming CHEN ; Yuhai GAO
Journal of Biomedical Engineering 2025;42(5):1054-1061
This study aims to investigate the therapeutic efficacy of 50 Hz-0.6 mT low-frequency pulsed electromagnetic field (PEMF) on postmenopausal osteoporosis in ovariectomized rats. Thirty 3-month-old female SD rats were selected and divided into a sham operation group (Sham), an ovariectomized model group (OVX), and a low-frequency pulsed electromagnetic field (PEMF) treatment group, with 10 rats in each group. After 8 weeks, the whole-body bone mineral density (BMD) of each group of rats was measured. The treatment group began to receive PEMF stimulation for 90 minutes daily, while the OVX group only received a simulated placement without electricity. After 6 weeks of intervention, all rats were sacrificed and tested for in vitro BMD, micro-CT, biomechanics, serum biochemical indicators, and bone tissue-related proteins. The results showed that the BMD of the OVX group was significantly lower than that of the Sham group 8 weeks after surgery, indicating successful modeling. After 6 weeks of treatment, compared with the OVX group, the PEMF group exhibited significantly increased BMD in the whole body, femur, and vertebral bodies. Micro-CT analysis results showed improved bone microstructure, significantly increased maximum load and bending strength of the femur, elevated levels of serum bone formation markers, and increased expression of osteogenic-related proteins. In conclusion, this study demonstrates that daily 90-minute exposure to 50 Hz-0.6 mT PEMF effectively enhances BMD, improves bone biomechanical properties, optimizes bone microstructure, stimulates bone formation, and inhibits bone resorption in ovariectomized rats, highlighting its therapeutic potential for postmenopausal osteoporosis.
Female
;
Animals
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Rats, Sprague-Dawley
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Osteoporosis, Postmenopausal/therapy*
;
Rats
;
Bone Density
;
Ovariectomy
;
Magnetic Field Therapy/methods*
;
Electromagnetic Fields
6.Prediction of early treatment outcomes in patients with traumatic brain injury using transcranial Doppler ultrasound combined with invasive intracranial pressure
Meng'ai ZHAI ; Li ZHANG ; Yang YANG ; Yuhai WANG ; Likun YANG ; Yang YANG
Journal of Army Medical University 2025;47(22):2783-2791
Objective To investigate the predictive role of transcranial Doppler(TCD)ultrasound combined with invasive intracranial pressure(ICP)monitoring in the early outcomes for patients with moderate-to-severe traumatic brain injury(msTBI)after ICP probe insertion surgery.Methods A retrospective case-control analysis was conducted on the clinical data of 65 msTBI patients who underwent invasive ICP probe insertion surgery within 24 h after injury and routine TCD examination postoperatively in No.904 Hospital of Joint Logistic Support Force of Chinese PLA between January 2021 and March 2025.Based on whether disease progression met the indications for craniotomy,they were divided into a failed conservative treatm ent group(n=28)and a successful conservative treatment group(n=37).Baseline clinical data,and parameters of TCD and ICP monitoring were collected in both groups.Univariate analysis was used for comparing differences in various parameters between the groups.The variables related to ICP and TCD were denoted as Model 1,and variables that significantly affected the outcome were incorporated as potential confounding factors into Model 2 based on clinical experience.Multivariate logistic regression analysis was performed to screen for independent predictors of conservative treatment failure.Receiver operating characteristic(ROC)curves were plotted to evaluate the diagnostic efficacy of each predictive factor.Results Statistical differences were observed between the 2 groups in terms of Glasgow Coma Scale(GCS)score at admission,postoperative mean intracranial pressure(ICPm),postoperative mean cerebral perfusion pressure(CPPm),and pulsatility index(PI)of the ipsilateral internal carotid artery extracranial segment(ICAt),and mean flow velocity(Vm)and pulsatility index(PI)of the ipsilateral middle cerebral artery(MCA)(all P<0.01).After adjusting for age,GCS score at admission,hematoma volum,and mean postoperative brain temperature,multivariate logistic regression analysis revealed that ICPm(OR=1.248,95%CI:1.044~1.491,P=0.015),ipsilateral MCA.Vm(OR=0.964,95%CI:0.933~0.997,P=0.034),and ipsilateral MCA.PI(OR=1.612,95%CI:1.026~2.533,P=0.038)were independent predictors of failure in conservative treatment in msTBI patients.After adjustment,ROC curve analysis demonstrated that the combined model of ipsilateral MCA.PI,ipsilateral MCA.Vm,and ICPm achieved a predictive efficacy of 0.931(95%CI:0.873~0.988),which was obviously higher than that of the single model of ICPm(0.875,95%CI:0.791~0.959).Conclusion Combined monitoring of TCD and ICP can provide a more accurate prediction of the early outcomes for msTBI patients after ICP probe insertion,providing a more effective assessment for clinical diagnosis and treatment.
7.Guideline for the diagnosis and treatment of vertebral refracture after percutaneous vertebral augmentation in elderly patients with osteoporotic thoracolumbar compression fractures (version 2025)
Yong YANG ; Xiaoguang ZHOU ; Qixin CHEN ; Jian CHEN ; Jian DONG ; Liangjie DU ; Shunwu FAN ; Jin FAN ; Zhong FANG ; Haoyu FENG ; Shiqing FENG ; Haishan GUAN ; Aiguo GAO ; Yanzheng GAO ; Yong HAI ; Da HE ; Dengwei HE ; Haiyi HE ; Dianming JIANG ; Xuewen KANG ; Bin LIN ; Baoge LIU ; Changqing LI ; Fang LI ; Li LI ; Fangcai LI ; Weishi LI ; Xiaoguang LIU ; Hongjian LIU ; Xinyu LIU ; Yong LIU ; Zhongjun LIU ; Shibao LU ; Xuhua LU ; Fei LUO ; Yuhai MA ; Keya MAO ; Xuexiao MA ; Bin MENG ; Xu NING ; Limin RONG ; Hongxun SANG ; Jun SHU ; Tiansheng SUN ; Dasheng TIAN ; Zheng WANG ; Bing WANG ; Linfeng WANG ; Qingde WANG ; Qinghe WANG ; Lan WEI ; Jigong WU ; Baoshan XU ; Youjia XU ; Guoyong YIN ; Jinglong YAN ; Feng YAN ; Cao YANG ; Huilin YANG ; Qiang YANG ; Bin ZHAO ; Jie ZHAO ; Yue ZHU ; Jianguo ZHANG ; Wenzhi ZHANG ; Zhongmin ZHANG ; Zhaomin ZHENG ; Yan ZENG ; Baorong HE ; Wei MEI
Chinese Journal of Trauma 2025;41(7):613-626
Vertebral refracture following percutaneous vertebral augmentation (PVA) is commonly seen in elderly patients with osteoporotic thoracolumbar compression fractures (OTLCF). It can lead to recurrent pain, loss of vertebral height, progression of kyphosis, and even neurological dysfunction, significantly impairing patients′ quality of life. Current diagnosis and treatment face multiple challenges, including high misdiagnosis rate, difficulty in choosing between surgical and non-surgical treatment options, lack of standardized surgical protocols, interference from intralesional bone cement during procedures, inadequate stability of internal fixation in osteoporotic bone, and suboptimal compliance of anti-osteoporotic therapy. Establishing a standardized diagnostic and therapeutic framework is urgently needed. To standardize the management process and improve outcomes for vertebral refractures after PVA in elderly OTLCF patients, Spinal Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized experts in the field to develop Guideline for the diagnosis and treatment of vertebral refracture after percutaneous vertebral augmentation in elderly patients with osteoporotic thoracolumbar compression fractures ( version 2025), based on current literature and clinical experience, and adhering to principles of scientific rigor and clinical applicability. A total of 11 recommendations were proposed, encompassing diagnosis, treatment, and rehabilitation of vertebral refracture after PVA in elderly patients with OTLCF, aiming to provide a foundation for a standardized management.
8.Construction and performance evaluation of a predictive model for post-traumatic hydrocephalus in patients with severe traumatic brain injury
Bin XU ; Xin WANG ; Jiahao LIAO ; Yuhai WANG ; Jinxu ZHOU
Chinese Journal of Trauma 2025;41(11):1059-1069
Objective:To develop a predictive model for the risk of post-traumatic hydrocephalus (PTH) in patients with severe traumatic brain injury (sTBI) and validate its predictive performance.Methods:A retrospective case control study was conducted to analyze the clinical data of 580 sTBI patients admitted to the 904th Hospital of the Joint Logistics Support Force of the PLA between January 2016 and December 2023, including 413 males and 167 females, aged 18-88 years [(54.3±14.6)years]. Patients were stratified into PTH group ( n=195) and non-PTH group ( n=385), based on the presence of PTH within 6 months after injury. Data collected from the two groups such as general baseline indicators, TBI-related clinical indicators (including surgical data), laboratory findings, and radiological features. Except for the data collected during the operation, all the above data are the results of the first examination at admission. Univariate analysis and Lasso regression analysis were used to screen predictors for the risk of PTH in sTBI patients. Subsequent multivariate Logistic regression was employed to identify predictors and construct a regression equation. Based on this equation, a nomogram prediction model was developed using the R language. Model discrimination was estimated through the receiver operating characteristic (ROC) curve, and calibration performance via the Hosmer-Lemeshow (H-L) goodness-of-fit test and calibration curve. Moreover, decision curve analysis (DCA) and clinical impact curve (CIC) were used for evaluating the clinical utility of the model. Results:Univariate analysis revealed statistically significant differences in 37 variables between the two groups, including age, age group, heart rate, oxygen saturation, Glasgow coma scale (GCS) score, left pupil size, right pupil size, pupillary light reflex, intracranial pressure (ICP) monitoring, type of decompressive craniectomy, neutrophil count, lymphocyte count, monocyte count, red blood cell count, platelet count, neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), neutrophil-lymphocyte-platelet ratio (N/LP), lymphocyte-to-monocyte ratio (LMR), systemic inflammation response index (SIRI), systemic immune-inflammation index (SII), fibrinogen (FIB), D-dimer, D-dimer-to-fibrinogen ratio (DFR), serum albumin, prognostic nutritional index, blood glucose, status of basal cisterns, midline shift, degree of midline shift, cerebral herniation, epidural hematoma (EDH), subdural hematoma (SDH), intraventricular hemorrhage (IVH), subarachnoid hemorrhage (SAH), modified Fisher grade, and skull fracture ( P<0.05). Lasso regression analysis identified 24 potential predictors for PTH, including age, GCS score, pupillary light reflex, type of decompressive craniectomy, monocyte count, platelet count, NLR, PLR, N/LP, LMR, SII, D-dimer, DFR, serum albumin, prognostic nutritional index, blood glucose, status of basal cisterns, degree of midline shift, cerebral herniation, EDH, SDH, IVH, modified Fisher grade and skull fracture. Multivariate Logistic regression analysis demonstrated that age, unilateral pupillary light reflex, absent pupillary light reflex, bilateral decompressive craniectomy, monocyte count, PLR, cerebral herniation, SDH, IVH, linear skull fracture and depressed skull fracture were independent risk factors for PTH. In contrast, serum albumin was identified as an independent protective factor for PTH ( P<0.05). The regression equation derived from these factors was: Logit[ P/(1- P)]=0.05×"age"+1.65×"unilateral pupillary light reflex"+2.79×"absent pupillary light reflex"+1.60×"bilateral decompressive craniectomy"+1.90×"monocyte count"+0.02×"PLR"-0.12×"serum albumin"+2.07×"cerebral herniation"+2.59×"SDH"+2.23×"IVH"+1.24×"linear skull fracture"+ 1.66×"depressed skull fracture"-22.61. The prediction model built upon this equation achieved an area under the ROC curve (AUC) of 0.95(95% CI 0.93, 0.97), with a sensitivity of 91.79%, specificity of 85.97%, and Youden′s index of 0.78. The H-L goodness-of-fit test indicated good calibration ( χ2=7.90, P=0.545). DCA results showed that the bias-corrected curve closely aligned with the actual curve and approximated the ideal curve, indicating a high clinical net benefit. Furthermore, CIC results demonstrated that with threshold probabilities greater than 60%, the number of patients identified as high-risk by the model highly corresponded with the actual number of patients who developed PTH. Conclusion:The prediction model incorporating age, unilateral pupillary light reflex, absent pupillary light reflex, bilateral decompressive craniectomy, monocyte count, PLR, serum albumin, cerebral herniation, SDH, IVH, linear skull fracture and depressed skull fracture exhibits robust predictive performance for PTH in sTBI patients.
9.Isopsoralen accelerates tibia fracture healing by promoting bone formation in mice
Xi WU ; Zhongqi WANG ; Yuhai GAO ; Zhenlong WEI ; Xin BAI ; Keming CHEN
Journal of Army Medical University 2024;46(3):240-248
Objective To determine the effect of isopsoralen(ISO)on the healing of tibia fracture in mice and explore its underlying mechanism.Methods Fifty male C57BL/6 mice(2 month old,20±2 g)were randomly divided into model group and ISO treatment group,with 25 animals in each group.From the 3rd day after modeling,the mice from the ISO group were given an intragastric gavage of 40 mg/kg ISO,once per day for 28 consecutive days,while those of the model group was given same volume of normal saline in same way.On the 7th,14th,21st,and 28th day after gavage,the tibia on the surgical side was taken,and the fracture area was quantified by bone volume/total volume(BV/TV)after micro-CT scanning.The healing and shaping of the fracture end were observed through HE staining.ELISA was used to detect the serum contents of bone alkaline phosphatase(BALP)and procollagen type I N-terminal peptide(PINP)on the 14th day of gavage.Western blotting was employed to determine the expression levels of Collagen Ⅰ,Runx2,BMP2,OSX,and VEGF in the tibial callus tissue in 7 and 14 d after gavage.Vascular perfusion was applied to observe the callus microvessels in 28 d to quantitatively analyze the vascular volume fraction and vessel diameter.Immunohistochemical staining was conducted to observe the expression of VEGF in the callus in 14 d after gavage.Results HE staining displayed that the ISO group had faster healing process than the model group.Micro-CT quantification results showed that the ISO group had higher BV/TV ratio in 7 d after gavage though no statistical difference,significantly higher ratio in 14 d(P<0.05),but obviously lower ratio in 21 and 28 d after gavage(both P<0.05)when compared with the model group.The serum contents of BALP and PINP were also remarkably higher in the ISO group than the model group(P<0.05).Western blotting results indicated that the expression levels of Collagen Ⅰ,Runx2,BMP2,OSX and VEGF in the ISO group were higher than those in the model group(P<0.05).The results of angiography revealed that the vascular volume fraction and vessel diameter were notably increased in the ISO group than the model group(both P<0.05).Immunohistochemical assay showed that the expression of VEGF was higher in the ISO group than the model group(P<0.05).Conclusion ISO can improve the activity of osteoblasts,increase the expression of osteogenesis-related proteins,and accelerate the angiogenesis to promote fracture healing.
10.Study on Isolation and Culture of Primary Lung Fibroblasts from Newborn Mice by Improved Tissue Adhesion Method
Xiaodan ZHENG ; Ting WANG ; Yuhai HU
Journal of Modern Laboratory Medicine 2024;39(6):206-210
Objective To establish an improved simple and efficient method for isolation and culture of primary lung fibroblasts(LFB)from neonatal mice,and study their growth characteristics in vitro.Methods The lungs of 3-days-old mice or 8-weeks-old mice were taken under sterile conditions,and the stromal tissue was cut into 1mm3 tissue mass.High-glucose DMEM culture medium containing 10g/dl fetal bovine serum(FBS)or 20g/dl FBS was used for tissue adhesion culture.The LFB was purified by the differential time adhesion method,and the growth morphology and adherence state of the cells were observed dynamically under an inverted microscope.The primary LFB was identified by flow cytometry.The activity of the third-generation cell after culture was detected by CCK-8 assay.Results Lung tissues in mice at 3 days of age and 20g/dl FBS concentration cultured by the improved tissue mass adherent cultured method began to grow radially to the periphery on the 2nd day.On the day 7th,the cells growth density reached 90%,and the cell morphology was as a spindle.The CD140a positive and CD45 negative cells reached more than 90%after purification by differential time adhesion method,and the cells still maintain good cell activity after cultured for 3 generations.Conclusion The improved tissue adhesion method can obtain a large number of purified mice LFB with good activity simply and efficiently,which lays a foundation for the study of lung inflammation,tumors and in vitro efficacy of drugs.


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