1.Ferroptosis and osteoporosis
Cheng YANG ; Weimin LI ; Dongcheng RAN ; Jiamu XU ; Wangxiang WU ; Jiafu XU ; Jingjing CHEN ; Guangfu JIANG ; Chunqing WANG
Chinese Journal of Tissue Engineering Research 2025;29(3):554-562
BACKGROUND:It has also been confirmed that ferroptosis is closely related to a variety of musculoskeletal diseases,such as rheumatoid arthritis,osteosarcoma,and osteoporosis.The pathophysiological mechanisms of ferroptosis and osteoporosis need to be further studied and elucidated to broaden our understanding of iron metabolism and osteoporosis.It will provide research ideas for the future elucidation of new mechanisms of osteoporosis and the development of new technologies and drugs for the treatment of osteoporosis. OBJECTIVE:To provide an overview of the current status of research on ferroptosis in osteoporosis,to provide a new direction for future research on the specific molecular mechanisms of osteoporosis,and to provide more effective and better options for osteoporosis treatment strategies. METHODS:The first author used the computer to search the literature published from 2000 to 2024 in CNKI,WanFang,VIP,and PubMed databases with search terms"ferroptosis,iron metabolism,osteoporosis,osteoblast,osteoclast,bone metabolism,signal pathway,musculoskeletal,review"in Chinese and English.A total of 68 articles were finally included according to the selection criteria. RESULTS AND CONCLUSION:(1)Ferroptosis is a new type of cell death discovered in recent years,which is usually accompanied by a large amount of iron accumulation and lipid peroxidation during cell death,and its occurrence is iron-dependent.This is distinctly different from several types of cell death that are currently being hotly studied(e.g.,cellular pyroptosis,necrotic apoptosis,cuproptosis,and autophagy).(2)Intracellular iron homeostasis is manifested as a balance between iron uptake,export,utilization,and storage.The body's iron regulatory system includes systemic and intracellular regulation.The main factor of systemic regulation is hepcidin produced by hepatic secretion,and cellular regulation depends on the iron regulatory protein/iron response element system.Of course,intracellular iron homeostasis can be controlled by other factors,such as hypoxia,cytokines,and hormones.(3)Lipid peroxidation causes oxidative damage to biological membranes(plasma membrane and internal organelle membranes),lipoproteins,and other lipid-containing molecules.Polyunsaturated fatty acid-containing phospholipids are important targets of lipid peroxidation.Free polyunsaturated fatty acid is an important substrate for lipid oxidation and can bind to the phospholipid bilayer,leading to over-oxidation and thus triggering lipid apoptosis.(4)Several studies have shown that osteoblasts are overloaded with iron in different ways,resulting in the accumulation of unstable ferrous iron and the generation of reactive oxygen species and lipid peroxides,causing ferroptosis of osteoblasts and ultimately a decrease in bone formation,affecting bone homeostasis and the development of osteoporosis.(5)Osteoclasts are large multinucleated cells formed by the fusion of mononuclear macrophage cell lines or bone marrow mesenchymal stem cells induced by nuclear factor-κB ligand receptor activator,and they have the function of bone resorption.Iron ions can promote osteoclast differentiation and bone resorption through the production of intracellular lipid reactive oxygen species,while iron chelators can inhibit osteoclast formation in vitro and thus affect the occurrence and development of osteoporosis.
2.Improvement effects and mechanism of urinastatin on cisplatin-induced myocardial injury
Jiafu REN ; Pengfei CHEN ; Rong A ; Jing LI
China Pharmacy 2025;36(8):920-925
OBJECTIVE To investigate the improvement effects and potential mechanism of ulinastatin (UTI) on cisplatin (CP)-induced myocardial injury. METHODS Rabbits were used as study subjects and randomly divided into blank group (normal saline 2 mL/d, for consecutive 7 d), CP group (150 mg/m2, on the 1st and 4th day), UTI low-dose/high-dose group (UTIL/UTIH group, UTL 25 000 or 50 000 IU/kg, once a day, for consecutive 7 d), c-Jun N-terminal kinase (JNK) inhibitor group (JNKi group, SP600125 15 mg/kg, on the 1st and 4th day+CP 150 mg/m2, on the 1st and 4th day+normal saline 2 mL/d, on the other 5 days). Except for the blank group, rabbits in the other groups were injected with relevant medicine and/or normal saline via a marginal ear vein on one side. The serum level of cardiac troponin Ⅰ (cTnⅠ) in rabbits on the 1st and 8th days of the detection experiment, as well as the levels of superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) in myocardial tissue on the 8th day of the experiment, were all measured in each group. The pathological changes and myocardial cell apoptosis in myocardial tissue were observed, and the protein expressions of B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), JNK, phosphorylated JNK (P-JNK), mitochondrial fission factor (Mff), and dynamin-related protein 1 (Drp1) in myocardial tissue were all determined. RESULTS Compared with the blank group, the CP group showed significant myocardial injury. The cell apoptotic rate, the levels of cTnⅠ in serum and MDA in myocardial tissue, as well as protein expressions of Bax, JNK, p-JNK, Mff, and Drp1, were all significantly increased or up-regulated, SOD level and protein expression of Bcl-2 significantly reduced or down+regulated(P<0.05). Compared with the CP group, the myocardial injury in rabbits from each drug treatment group showed improvement. The cell apoptosis index, the levels of cTnⅠ in serum and MDA in myocardial tissue, as well as protein expressions of Bax, JNK, p-JNK, Mff (except for UTIL group), and Drp1, were all significantly reduced or down-regulated, while SOD, GSH level and protein expression of Bcl-2 significantly increased or up-regulated (P<0.05); moreover, the improvement in some indicators was significantly better in the UTIH group and the JNKi group compared to the UTIL group (P<0.05). CONCLUSIONS UTI may ameliorate CP-induced myocardial injury, the potential mechanism of which may be associated with antagonizing oxidative stress and inhibiting the JNK/Mff signaling pathway.
3.Restoration of a dental defect in a patient with ectodermal dysplasia: a case report and literature review
LV Hong ; LIU Qinghui ; LI Jiafu ; CAI Hui ; BU Honghu ; PU Yiming ; GUO Jincai
Journal of Prevention and Treatment for Stomatological Diseases 2024;32(4):287-295
Objective:
To explore the treatment options for congenitally missing teeth in patients with ectodermal dysplasia and provide a clinical reference.
Methods:
A patient with ectodermal dysplasia with a concave midface, anterior protrusion of the chin, and underdevelopment of the lower third of the face presented with congenital loss of multiple maxillary teeth, malocclusion of the remaining teeth, congenital loss of mandibular dentition, small dental arches, and upper and lower alveolar bone hypoplasia. The patient was treated by means of a removable partial maxillary prosthesis, implants in the anterior region of the lower mandible designed with the assistance of digital guides, and bar-clamped implant-overlay prostheses. A literature review of the protocol for the treatment of this condition was also conducted.
Results:
In addition to good retention and stability after denture wear, an excellent occlusal relationship, improvement of the patient's facial appearance, including upper and lower lip fullness, more equal balancing of the lower and middle 1/3 of the face, and improved masticatory function were achieved. The results of the literature review showed that patients with ectodermal dysplasia who are congenitally edentulous usually have a complex intraoral situation that makes restoration difficult, and common restorative modalities for these patients include fixed bridges, removable partial dentures, complete dentures, overdentures, and implant prostheses, which need to be selected according to the actual intraoral situation of each patient. Currently, there is no consensus on the treatment of congenitally missing teeth in patients with ectodermal dysplasia, and some scholars have suggested that fixed restorations be recommended for patients with fewer missing teeth, while the option of removable or implant-covered denture restorations should be given to patients with more missing teeth, with removeable prostheses for underage patients that are replaced with permanent fixed prostheses when the jaws have stabilized.
Conclusion
In patients with ectodermal dysplasia with congenital tooth loss, all factors should be taken into account, and an individualized restorative plan should be developed.
4.Prognosis and influencing factors analysis of patients with initially resectable gastric cancer liver metastasis who were treated by different modalities: a nationwide, multicenter clinical study
Li LI ; Yunhe GAO ; Liang SHANG ; Zhaoqing TANG ; Kan XUE ; Jiang YU ; Yanrui LIANG ; Zirui HE ; Bin KE ; Hualong ZHENG ; Hua HUANG ; Jianping XIONG ; Zhongyuan HE ; Jiyang LI ; Tingting LU ; Qiying SONG ; Shihe LIU ; Hongqing XI ; Yun TANG ; Zhi QIAO ; Han LIANG ; Jiafu JI ; Lin CHEN
Chinese Journal of Digestive Surgery 2024;23(1):114-124
Objective:To investigate the prognosis of patients with initially resectable gastric cancer liver metastasis (GCLM) who were treated by different modalities, and analyze the influencing factors for prognosis of patients.Methods:The retrospective cohort study was conducted. The clinicopathological data of 327 patients with initially resectable GCLM who were included in the database of a nationwide multicenter retrospective cohort study on GCLM based on real-world data from January 2010 to December 2019 were collected. There were 267 males and 60 females, aged 61(54,68)years. According to the specific situations of patients, treatment modalities included radical surgery combined with systemic treatment, palliative surgery combined with systemic treatment, and systemic treatment alone. Observation indicators: (1) clinical characteristics of patients who were treated by different modalities; (2) prognostic outcomes of patients who were treated by different modalities; (3) analysis of influencing factors for prognosis of patients with initially resectable GCLM; (4) screening of potential beneficiaries in patients who were treated by radical surgery plus systemic treatment and patients who were treated by palliative surgery plus systemic treatment. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the rank sum test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. The Kaplan-Meier method was used to calculate survival rate and draw survival curve, and Log-Rank test was used for survival analysis. Univariate and multivariate analyses were conducted using the COX proportional hazard regression model. The propensity score matching was employed by the 1:1 nearest neighbor matching method with a caliper value of 0.1. The forest plots were utilized to evaluate potential benefits of diverse surgical combined with systemic treatments within the population. Results:(1) Clinical characteristics of patients who were treated by different modalities. Of 327 patients, there were 118 cases undergoing radical surgery plus systemic treatment, 164 cases undergoing palliative surgery plus systemic treatment, and 45 cases undergoing systemic treatment alone. There were significant differences in smoking, drinking, site of primary gastric tumor, diameter of primary gastric tumor, site of liver metastasis, and metastatic interval among the three groups of patients ( P<0.05). (2) Prognostic outcomes of patients who were treated by different modalities. The median overall survival time of the 327 pati-ents was 19.9 months (95% confidence interval as 14.9-24.9 months), with 1-, 3-year overall survival rate of 61.3%, 32.7%, respectively. The 1-year overall survival rates of patients undergoing radical surgery plus systemic treatment, palliative surgery plus systemic treatment and systemic treatment alone were 68.3%, 63.1%, 30.6%, and the 3-year overall survival rates were 41.1%, 29.9%, 11.9%, showing a significant difference in overall survival rate among the three groups of patients ( χ2=19.46, P<0.05). Results of further analysis showed that there was a significant difference in overall survival rate between patients undergoing radical surgery plus systemic treatment and patients undergoing systemic treatment alone ( hazard ratio=0.40, 95% confidence interval as 0.26-0.61, P<0.05), between patients undergoing palliative surgery plus systemic treatment and patients under-going systemic treatment alone ( hazard ratio=0.47, 95% confidence interval as 0.32-0.71, P<0.05). (3) Analysis of influencing factors for prognosis of patients with initially resectable GCLM. Results of multivariate analysis showed that the larger primary gastric tumor, poorly differentiated tumor, larger liver metastasis, multiple hepatic metastases were independent risk factors for prognosis of patients with initially resectable GCLM ( hazard ratio=1.20, 1.70, 1.20, 2.06, 95% confidence interval as 1.14-1.27, 1.25-2.31, 1.04-1.42, 1.45-2.92, P<0.05) and immunotherapy or targeted therapy, the treatment modality of radical or palliative surgery plus systemic therapy were independent protective factors for prognosis of patients with initially resectable GCLM ( hazard ratio=0.60, 0.39, 0.46, 95% confidence interval as 0.42-0.87, 0.25-0.60, 0.30-0.70, P<0.05). (4) Screening of potentinal beneficiaries in patients who were treated by radical surgery plus systemic treatment and patients who were treated by palliative surgery plus systemic treatment. Results of forest plots analysis showed that for patients with high-moderate differentiated GCLM and patients with liver metastasis located in the left liver, the overall survival rate of patients undergoing radical surgery plus systemic treatment was better than patients undergoing palliative surgery plus systemic treatment ( hazard ratio=0.21, 0.42, 95% confidence interval as 0.09-0.48, 0.23-0.78, P<0.05). Conclusions:Compared to systemic therapy alone, both radical and palliative surgery plus systemic therapy can improve the pro-gnosis of patients with initially resectable GCLM. The larger primary gastric tumor, poorly differen-tiated tumor, larger liver metastasis, multiple hepatic metastases are independent risk factors for prognosis of patients with initial resectable GCLM and immunotherapy or targeted therapy, the treatment modality of radical or palliative surgery plus systemic therapy are independent protective factors for prognosis of patients with initially resectable GCLM.
5.Pregnancy with recurrent endometrial stromal sarcoma: a case report
Jinghong JIANG ; Jing CHENG ; Xiaoyan HE ; Jiafu LI ; Kai ZHANG
Chinese Journal of Perinatal Medicine 2024;27(3):233-237
This paper reported the clinical characteristics, diagnosis, and treatment of a case of recurrent endometrial stromal sarcoma with term pregnancy. The patient had undergone laparoscopic surgery to remove hysteromyoma before conception in 2017, which was pathologically diagnosed as low-grade endometrial stromal sarcoma after surgery. Due to her strong reproductive willingness, the patient attempted to conceive in light of her stable condition and no evidence of recurrence and was closely followed up with an informed choice. She conceived successfully in 2020 and underwent regular pregnancy examinations. Ultrasound examination at 37 +4 weeks of gestation revealed a slightly hypoechoic mass of about 6.3 cm×4.5 cm size in the pelvic cavity. After admission, a pelvic MRI indicated multiple solid nodules in the right adnexa uteri and beside the iliac vessels in the left pelvic wall and anterior pelvic wall with the larger one being about 58 mm×28 mm. Diffusion-weighted imaging showed multiple pelvic nodules and masses with significant diffusion restriction. The patient was diagnosed as having multiple solid nodules and masses in the pelvic cavity, and the recurrence of sarcoma was highly suspected. Brain CT and lung CT showed no obvious metastatic lesions. A consultation involving the Department of Gynecological Tumor Chemoradiotherapy was held and the sarcoma recurrence during pregnancy was prenatally diagnosed. After ruling out the contraindications for surgery, a cesarean section was performed in the lower segment of uterus under general anesthesia and a live female baby was delivered at 38 +3 weeks. The excised mass was confirmed as recurrent uterine stromal sarcoma by rapid freezing pathology during cesarean section. A combination surgery was performed subsequently, including total extra-fascial hysterectomy, bilateral oophorectomy, bilateral salpingectomy, appendectomy, greater omentum resection, pelvic lesion resection (right side), and pelvic adhesiolysis. Recurrent low-grade uterine stromal sarcoma was reconfirmed by postoperative pathology. The patient was discharged after recovery. After two years of follow-up, no distant metastasis recurrences were found.
6.Interpretation on the report of global cancer statistics 2022
Xi ZHANG ; Lei YANG ; Shuo LIU ; Lili CAO ; Ning WANG ; Huichao LI ; Jiafu JI
Chinese Journal of Oncology 2024;46(7):710-721
In April 2024, the World Health Organization/International Agency for Research on Cancer (IARC) published the global cancer statistics 2022 in the CA: Cancer Journal for Clinicians. This report focuses on the incidence and mortality of 36 cancers in 185 countries or territories worldwide, analyzing the differences of gender, geographic region, and the Human Development Index (HDI) level. It is estimated that in the year 2022, there were 19.96 million new cancer cases and 9.74 million cancer deaths worldwide. Lung cancer (2 480 301, 12.4%) was the most frequently diagnosed cancer in 2022, followed by female breast cancer (2 295 686, 11.5%), colorectal cancer (1 926 118, 9.6%), prostate cancer (1 466 680, 7.3%), and gastric cancer (968 350, 4.9%). Lung cancer (1 817 172, 18.7%) was also the leading cause of cancer death, followed by colorectal cancer (903 859, 9.3%), liver cancer (757 948, 7.8%), female breast cancer (665 684, 6.9%), and gastric cancer (659 853, 6.8%). With demographics-based predictions indicating that the number of new cases of cancer will reach over 35 million by 2050. The Beijing Office for Cancer Prevention and Control team has collated this report and briefly interpreted it in combination with the current situation of cancer incidence and mortality in China.
7.Chinese national clinical practice guidelines on the prevention, diagnosis, and treatment of early gastric cancer
Peng LI ; Ziyu LI ; Enqiang LINGHU ; Jiafu JI
Chinese Medical Journal 2024;137(8):887-908
Background::Gastric cancer is one of the most common malignant tumors in the digestive system in China. Few comprehensive practice guidelines for early gastric cancer in China are currently available. Therefore, we created the Chinese national clinical practice guideline for the prevention, diagnosis, and treatment of early gastric cancer.Methods::This clinical practice guideline (CPG) was developed in accordance with the World Health Organization’s recommended process and with the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) in assessing evidence quality. We used the Evidence to Decision framework to formulate clinical recommendations to minimize bias and increase transparency in the CPG development process. We used the Reporting Items for practice Guidelines in HealThcare (RIGHT) statement and the Appraisal of Guidelines for Research and Evaluation II (AGREE II) as reporting and conduct guidelines to ensure completeness and transparency of the CPG.Results::This CPG contains 40 recommendations regarding the prevention, screening, diagnosis, treatment, and follow-up of early gastric cancer based on available clinical studies and guidelines. We provide recommendations for the timing of Helicobacter pylori eradication, screening populations for early gastric cancer, indications for endoscopic resection and surgical gastrectomy, follow-up interval after treatment, and other recommendations. Conclusions::This CPG can lead to optimum care for patients and populations by providing up-to-date medical information. We intend this CPG for widespread adoption to increase the standard of prevention, screening, diagnosis, treatment, and follow-up of early gastric cancer; thereby, contributing to improving national health care and patient quality of life.
8.Comparison of clinical efficacies among three surgical methods in treating stage Ⅲ Kümmell disease without neurological symptoms
Xudong MENG ; Jiongbiao ZHONG ; Chao DONG ; Yin XU ; Jiafu LI
Chongqing Medicine 2024;53(16):2512-2519
Objective To compare the clinical efficacies among three surgical methods in the treatment of stage Ⅲ Kümmell's disease without neurological symptoms.Methods A total of 71 patients with stage Ⅲ Kümmell's disease without neurological symptoms admitted and treated in the orthopedics department of Af-filiated Yueyang Hospital of Hunan Normal University from September 2019 to April 2022 were selected and divided into three groups:29 cases in the percutaneous vertebroplasty (PVP) group (group A),19 cases in the percutaneous pedicle screw+lesioned vertebral PVP group (group B) and 21 cases in the percutaneous bone cement screw combined with lesioned vertebral PVP group (group C) according to the operation methods.The imaging,perioperative and follow-up indexes were compared among the three groups.Results All patients successfully completed the operation,no serious complications occurred after operation,and the follow-up time was more than 1 year.The sagittal position index (SI value) in the affected vertebra,visual analogue scale (VAS) score and Oswestry Disability Index (ODI) score at each time point (postoperative 1,3 months,1 year) after surgery in the group A were improved compared with before operation (P<0.05).The affected vertebra SI value,kyphosis Cobb angle,spinal canal stenosis rate,VAS score and ODI score at postoperative time points in the group B and C were improved compared with before operation (P<0.05).The affected ver-tebra SI value,kyphosis Cobb angle,spinal canal stenosis rate and ODI score at postoperative time points in the group B and C all were superior to those in the group A (P<0.05),while the affected vertebra SI value,kyphosis Cobb angle and spinal canal stenosis rate in postoperative 1 year in the group C were superior to those in the group B (P<0.05).The operative time,intraoperative bleeding volume,postoperative hospitaliza-tion duration and affected vertebral cement injection volume in the group A were significantly less than those in the group B and C (P<0.05).The operation time in the group B was less than that in the group C (P<0.05).Conclusion The three surgical methods for the treatment of stage Ⅲ Kümmell's disease without neu-rological symptoms all could relieve the symptoms and restore the function.The two surgical methods of pedi-cle screw reduction could more effectively restore the height of the affected vertebra,correct kyphosis malfor-mation and improve spinal stenosis than PVP alone.The pedicle screw has more advantages in maintaining mechanical support and stability after bone cement reinforcement.
9.Construction and evaluation of X-ray induced senescence model of rat intervertebral disc nucleus pulposus cells in vitro
Jiongbiao ZHONG ; Jiafu LI ; Vo NAM
Chinese Journal of Spine and Spinal Cord 2024;34(10):1068-1076
Objechves:To establish an X-ray induced senescence model of rat intervertebral disc nucleus pulposus cells in vitro and to evaluate it.Methods:12 clean-grade mature SD rats were selected to take the lumbar intervertebral disc nucleus pulposus for cell culture in vitro,and the cultured nucleus pulposus cells were randomly divided into 0 Gy group(Control group),5Gy group,10Gy group and 15Gy group on the basis of different X-ray doses of irridiation.Cell morphology observation,SA-β-Gal staining,cell counting kit-8(CCK-8),flow cytometry(FCM),polymerase chain reaction(PCR),immunofluorescence(IF)technique,Western blot(WB)and 1,9-dimethyl-methylene blue(DMMB)colorimetric method were used to observe and compare the senescence and proliferation of different groups of nucleus pulposum cells.The expression of age-related genes,proteins and related factors were detected.Results:The nucleus pulposus cells gradually showed typi-cal characteristics of senescence in morphology with the increase of X-ray radiation dose,and the degree of aging was positively correlated with radiation dose.SA-β-Gal assay showed that the positive percentages of nucleus pulposus cells in 0Gy,5Gy,10Gy and 15Gy groups were 15%,41%,88%and 77%,respectively.CCK-8 experiment showed on the 5th day,the proliferation of nucleus pulposus cells in the 10Gy group re-sumed,while that in the 15Gy group still showed no proliferation.FCM confirmed that the cells were still active,and when the radiation dose was increased to 20Gy,the early apoptosis rate of nucleus pulposus cells increased sharply.Both IF and PCR showed that the expression of aging genes p16 and p21 increased with the increase of radiation dose,and the expression of aggrecan fragmentation was also increased in WB assay.The above experimental results suggested that 15Gy was the appropriate radiation dose.When the 15Gy group was compared with the control group again,CCK-8 showed no proliferation of nucleus pulposus cells in the 15Gy group on day 9,and the PCR showed significantly higher expression of aggrecan fragmentation in the 15Gy group than the control group(P<0.05).The expression of MMP-1,MMP-3 and inflammatory factors IL-6 and IL-8 in PCR and IF were also significantly higher in 15Gy group than in the control group(P<0.05).Conclusions:X-ray can induce senescence of the rat nucleus pulposus cells cultured in vitro,and the senescence degree of nucleus pulposus cells receiving 15Gy radiation is obvious and stable,which can be used as an ideal model of nucleus pulposus cell senescence.
10.Sequencing analysis of whole genome of one strain of bovine Akabane virus and es-tablishment of fluorescence quantitative PCR for virus detection
Jiafu SHANG ; Le HU ; Mingke LI ; Yujian WU ; Xingwei NI ; Xiaowei YANG ; Xia LIU ; Liwu ZHANG ; Tingting XU ; Guangwei ZHAO
Chinese Journal of Veterinary Science 2024;44(9):1872-1881
In order to understand the prevalence of Akabane disease(AKAD)in Guizhou Province and the molecular characteristics of the isolates,the whole-genome sequence of a strain of Akabane virus(AKAV)from a bovine AKAD-positive sample was determined and analyzed.The genotype and genetic variation of the strain were also explored.Based on the conserved S sequence,a fluores-cence quantitative PCR(qPCR)detection method was established and applied for the investigation of AKAV infection status in four large-scale beef cattle farms of Guizhou.Results showed that the S,M and L fragments of the bovine strain were highly homologous to the Tianjin strain(TJ2016/China/2016)and the Australian strain(JaLAB39/Australia/1959),where they were in the same evolutionary branch and belonged to genotype Ⅱ.Sensitivity assay found that the lowest detection limit was 2.5 X 101 copies/μL.Specificity assay showed the established method detected only AKAV with no amplification on bovine bluetongue virus(BLUV),Pasteurella multocida(PM),bovine infectious rhinotracheitis virus(IBRV)and bovine Mycoplasma bovis.The variation coefficients of inter-and intra batches in the repeatability test were both lower than 2.26%.These findings illus-trated that the established qPCR method had high sensitivity,good specificity and repeatability.A total of 298 serum samples from 4 large-scale beef cattle farms in Qianxi City and Huangping County of Guizhou Province were collected and tested for AKAV by the method.Out of 298 sam-ples,25 positive samples(25/298)were detected as positive with a positive rate of 8.39%.In sum-mary,this work provided the reference data for a deep understanding of the molecular prevalence of AKAV in Guizhou Province and laid foundation for the prevention and control of AKAD.


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