1.Exploration and Practice of Safe Access System Construction for Barrier Environment Facilities of Laboratory Animals: A Case Study on Xianlin Campus of Nanjing University
Dongxia HOU ; Zuoxiu TIE ; Yong LU ; Panpan NAN ; Jie BAO
Laboratory Animal and Comparative Medicine 2025;45(1):96-100
Laboratory animals are essential in scientific research and experimental teaching in colleges and universities. Disciplines such as life sciences, medicine, pharmacy, chemistry, and biomedical engineering heavily rely on animal experiments. The standardized barrier environmental facility for laboratory animals provides a fundamental platform for stable, scientific, and reliable animal experiment results. Rigorous access management for such facilities is a vital safeguard for maintaining standardized operations of facilities, controlling the quality and stability of laboratory animals, mitigating pathogen contamination risks among animals and laboratory staff, and preventing biosecurity incidents such as zoonotic disease outbreaks. Taking the small-scale barrier facilities for laboratory rats and mice at Nanjing University's Xianlin Campus, operational since 2019, as an example, this study focuses on the safety access management system of these facilities. Based on five years of operational data and accumulated experience in studying and optimizing the access management system, this study, from the perspectives of management system development and the formulation and implementation of standard operating procedures, reviews five aspects of access management: personnel access, animals access, material access, equipment access, and air circulation control. Furthermore, these aspects are systematically analyzed and summarized to serve as a reference for the construction and management of the laboratory animal facilities in universities, while also contributing to scientific research, public health security, and the well-being of experimental personnel.
2.Effects of supernatant of BV-2 cells induced by LPS on inflammatory response and apoptosis in HT22 neurons
Li-ya WU ; Xin-ru WANG ; Yu-jie WU ; Wei-yi ZHANG ; Nan LI ; Yong-hui WANG ; Li GAO ; Le ZHAO
Chinese Pharmacological Bulletin 2025;41(7):1324-1331
Aim To observe the effect of lipopolysac-charide(LPS)induced supernatant of BV-2 cells on the inflammatory response and apoptosis of HT22 neu-rons.Methods After the concentration and time of LPS were determined by CCK-8 method,BV-2 cells were cultured with medium without LPS and medium containing LPS,the morphological changes of BV-2 microglia were observed by inverted microscope,and the CD86/CD206 ratio of BV-2 microglia was detected by immunofluorescence.Subsequently,BV-2 cell cul-ture supernatants were isolated and added to HT22 neuronal culture to observe the effect on the inflamma-tory response of HT22 neurons.The proliferation of HT22 neurons was detected by CCK-8 method and EdU method.The structural changes of HT22 neurons were observed under the microscope and examined by urani-um-lead staining.The levels of cytokines interleukin-1β(IL-1β),interleukin-10(IL-10),nuclear factor kappa-B(NF-κB)and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent as-say(Elisa).Neuronal apoptosis was detected by the TUNEL method.The protein expressions of Bax,Bcl-2 and inflammatory factors were detected by Western blot.Results After induction with 1 mg·L-1 LPS,BV-2 cells exhibited increased cell body size,thicker protrusions on both side,and some cells showed de-formed protrusions,the CD86/CD206 ratio in BV-2 cells decreased,promoting the transformation of BV-2 cells from M2 type to M1 type.After treating with the culture supernatant of BV-2 cells,HT22 neuronal cell activity and proliferation were reduced,axons short-ened,and the number of cells decreased.Neuronal cell bodies were enlarged and some cells were de-formed,with damaged cell membranes,round cell nu-clei but displaced nucleoli from the normal position,swollen mitochondria with vacuoles,reduced internal ridge structures,and increased levels of inflammatory factors NF-κB,IL-1 β,and TNF-α(P<0.05 or P<0.01),while the anti-inflammatory factor IL-10 de-creased(P<0.05),protein expression of the pro-apoptotic indicator Bax increased(P<0.01),and the protein expression of the anti-apoptotic indicator Bcl-2 decreased(P<0.05).Conclusion After induction of BV-2 cell polarization by LPS,the supernatant could inhibit HT22 neuronal cell viability,upregulate inflam-matory factor expression and promote apoptosis.
3.Cloning,bioinformatics analysis,expression and localization of APOD in bactrian camel epididymis
Aili CUI ; Wenjing WANG ; Xue HUANG ; Qiu YAN ; Tianan LI ; Jinghong NAN ; Yong ZHANG ; Xingxu ZHAO ; Qi WANG
Chinese Journal of Veterinary Science 2025;45(4):752-759
Apolipoprotein D(APOD)is a protein that is widely present in animal tissues and is in-volved in the reproductive regulation of the body.In order to investigate the expression regularity of APOD in bactrian camel epididymis and its regulation effect on sperm maturation,this study took the epididymis of bactrian camel during estrus and anestrus as materials,and first cloned the complete sequence of APOD coding sequence(CDS)region.The physicochemical properties of AP-OD were analyzed by ProParam,SOPMA,SWISS-MODEL and MEGA7.0 software.Meanwhile,the expression and distribution of APOD in epididymis were detected by qRT-PCR,Western blot and IHC.The cloning results showed that:the length of the CDS region of APOD gene was 624 bp,encoding 207 amino acids.The APOD sequence of Bactrian camel was highly conserved with the nucleotide and amino acid sequence of alpaca,and the homology of APOD sequence with elk was the lowest.The results of qRT-PCR showed that the mRNA levels of APOD in the head,body and tail of epididymis in estrus were significantly higher than those in estrus(P<0.01).Western blot results showed that the APOD protein expression and mRNA expression trend was similar in the head and body of the epididymis during anestrus,but the APOD expression level in the tail of the epididymis during anestrus was opposite to the mRNA expression level(P<0.05).The results of H&E and IHC showed that there were significant differences in epididymal tissue between estrus and anestrus.In addition,APOD showed positive reactions in epididymal epithelial cells,smooth muscle cells,sperm and connective tissue to varying degrees,suggesting that APOD may be in-volved in the maturation of sperm during estrus and anestrus,providing evidence for further explo-ring the regulatory mechanism of APOD's involvement in seasonal estrus.
4.Analysis of the impact of phase Ⅱ cardiac rehabilitation on the rehabilitation needs and physical activity of patients with coronary heart disease after interventional surgery
Ying ZHANG ; Xiwei ZHANG ; Jiahui WU ; Wenxian LIU ; Yan FENG ; Nan LI ; Yong SHAO ; Lixin ZHANG ; Yuan TIAN ; Zhiying LI ; Yin LI
Chongqing Medicine 2025;54(5):1080-1084
Objective To investigate the impact of phase Ⅱ cardiac rehabilitation on the rehabilitation needs and physical activity status of patients after coronary heart disease intervention.Methods A total of 90 patients with coronary heart disease who underwent percutaneous coronary intervention(PCI)in the Coronary Heart Disease Center of the hospital from August 2023 to August 2024 were selected as the research subjects.They were subjected to a 12-week standardized phase Ⅱ cardiac rehabilitation training.General data survey forms,cardiac rehabilitation scales,and the International Physical Activity Questionnaire were used for scale surveys to understand the patients' needs before and after rehabilitation and their weekly physical activity en-ergy expenditure.The cardiopulmonary exercise test gold standard,which reflects exercise capacity through three indicators-maximum oxygen uptake(VO2 max),anaerobic threshold(AT),and metabolic equivalents(MET),were used to compare the physical activity status before and after cardiac rehabilitation.Results Compared with before the implementation,after the implementation of cardiac rehabilitation,the autonomy score in-creased(21.36±1.85 vs.16.73±3.28),the process anxiety(12.60±3.87 vs.14.27±2.12)and outcome anxiety scores(2.31±1.76 vs.4.56±3.56)decreased,the level of low-intensity physical activity decreased[(2 711.62±1 487.09)min/week vs.(3 845.97±2 083.71)min/week],the levels of moderate-intensity[(1 314.67±783.54)min/week vs.(686.22±126.79)min/week],high-intensity[(1 861.33±798.27)min/week vs.(112.00±40.77)min/week],and total physical activity increased[(5 887.62±2 843.54)min/week vs.(4 644.19±2 287.16)min/week].The levels of VO2 max[(28.11±14.28)mL·min-1·kg-1 vs.(23.82±12.34)mL·min-1·kg-1],AT[(16.06±5.41)mL·min-1·kg-1 vs.(13.53±4.56)mL·min-1·kg-1],and MET[(6.89±1.59)mL·min-1·kg-1 vs.(5.78±1.21)mL·min-1·kg-1]all in-creased,with statistically significant differences(P<0.05).Conclusion Phase Ⅱ rehabilitation after PCI can effectively improve patients' physical activity levels.
5.Research and Application Progress of Polymer Magnetic Microspheres in the Fields of Biomedicine and Medical Diagnosis
Yong-Shuai TIAN ; Han WANG ; Fan YANG ; Jia-Nan LIU ; Huan-Lei WANG
Chinese Journal of Analytical Chemistry 2025;53(5):717-728
Polymer magnetic microspheres are a new type of composite functional materials,which have the advantages of stable structure,diverse modification methods,and magnetic response.These characteristics make polymer magnetic microspheres have the advantages of high sensitivity,high-specificity and fast response when detecting logo carriers or modified materials,and shows great application prospects in the fields of biomedicine and medical diagnosis.In this review,the basic properties of polymer magnetic microspheres were summarized,the research progresses of magnetic microspheres in the fields of biomedicine and medical diagnosis were emphatically introduced,and the improvement of preparation methods and development trends in the future were prospected.
6.Multi-center clinical study on the efficacy and safety of combined lienal polypeptide injection therapy in children with Mycoplasma pneumoniae pneumonia
Qi CHENG ; Yunxiao SHANG ; Han ZHANG ; Jiujun LI ; Ning CHEN ; Lishen SHAN ; Nan YANG ; Lihua NING ; Xuemei BAI ; Jianhua LIU ; Yuling HAN ; Jichun WANG ; Jing LI ; Yong FENG ; Liyun LIU ; Li CHEN ; Si LIU ; Qinzhen ZHANG ; Jia ZHENG ; Fengchao LI ; Sukun LU ; Yun ZHANG ; Xiaoyi CHAI
International Journal of Pediatrics 2025;52(3):204-210
Objective:To observe the efficacy and safety of combined lienal polypeptide injection therapy in the treatment of Mycoplasma pneumoniae pneumonia(MPP)in children aged 3 to 14 years old in multiple clinical centers.Methods:A randomized,controlled,multi-center clinical study design was adopted.A total of 240 hospitalized children aged 3 to 14 years old with MPP from 7 hospitals from September 1,2023 to January 31,2024 were included.According to the severity of pneumonia,they were divided into the mild MPP group with 80 cases and the severe MPP/refractory MPP(SMPP/RMPP)group with 160 cases,and then randomly divided into the control group and the experimental group at a ratio of 1 ∶1,using the random number table method.After screening,subjects entered a treatment period of 5 to 7 days.The control group was treated with azithromycin,while the experimental group was treated with azithromycin plus lienal polypeptide injection .The recovery of lung CT,length of hospital stay,duration of fever,cough score,whether mild cases developed into severe or refractory cases,duration of hormone use,use of intravenous immunoglobulin(IVIG),bronchoscopy treatment,and immune function were observed between the two groups to evaluate the efficacy of lienal polypeptide injection.Adverse events after medication,vital signs,blood routine,urine routine,liver function,myocardial enzymes,renal function,and electrocardiogram were observed to evaluate the safety. Results:A total of 231 subjects have completed the trial in the 7 hospitals,including 118 cases in the experimental group and 113 cases in the control group.Main observation index:the rate of lung CT aggravation in the experimental group was lower than that in the control group(2.6% vs 15.3%, P<0.01),and the difference was statistically significant.Secondary indexes:there were no statistically significant differences in the length of hospital stay,duration of fever,cough score,duration of hormone use,whether IVIG treatment was used,the number of bronchoscopy treatment cases,and immunoglobulin between the two groups(all P>0.05).However,the rate of cases of plastic bronchitis(PB)found under bronchoscopy in the experimental group was lower than that in the control group(0 vs 18.8%, P=0.03),and the difference was statistically significant.Among the mild MPP(72 cases),there were no statistically significant differences in the length of hospital stay,duration of fever,cough score,duration of hormone use,whether IVIG treatment was used,the number of bronchoscopy treatment cases,and the improvement rate of lung CT between the two groups(all P>0.05).However,compared with the control group,the rate of cases developing into SMPP/RMPP in the experimental group was less(24.3% vs 48.6%, P=0.03),and the difference in IgG before and after treatment was small[0.53(-0.04,1.18)g/L vs 1.33(0.48,2.25)g/L, P=0.01].Among the SMPP/RMPP cases(159 cases),the rate of cases of PB found under bronchoscopy in the experimental group was less than that in the control group(0 vs 20%, P=0.04),and the rate of cases with aggravated lung CT in the experimental group was less than that in the control group(1.3% vs 19.5%, P<0.01),and the improvement rate of lung CT in the experimental group was higher than that in the control group(88.8% vs 75.3%, P=0.03),with statistically significant differences.There were no statistically significant differences in the length of hospital stay,duration of fever,cough score,duration of hormone use,whether IVIG treatment was used,the number of bronchoscopy treatment cases,and immunoglobulin between the two groups(all P>0.05).Two cases in the experimental group developed rashes,which improved after the drug was discontinued.There were no serious adverse reactions such as abnormal vital signs like dyspnea and cyanosis due to the use of lienal polypeptide injection.There were no obvious changes in blood routine,liver function,myocardial enzymes,renal function,electrocardiogram,and urine routine values before and after medication compared with the baseline. Conclusion:The combined use of lienal polypeptide injection in the treatment of MPP in children can reduce the probability of the transformation from mild cases to SMPP/RMPP,reduce the rate of aggravation of the image findings,promote the absorption of lung inflammation,reduce the rate of PB found under bronchoscopy,and has good safety.
7.Observation of patient clinical outcomes based on the new subclassification of zone Ⅲ open-globe injury
Peng GU ; Sha LI ; Dianjun JIANG ; Yong LIU ; Nan WU
Chinese Journal of Ocular Fundus Diseases 2025;41(4):271-276
Objective:To compare and observe the visual acuity and ocular anatomical outcome of different subtypes in open-globe injury (OGI) Ⅲ.Methods:A retrospective study. A total of 187 eyes of 187 patients with OGI involving zone Ⅲ who were admitted to the Department of Ophthalmology of The First Affiliated Hospital of Army Medical University from January 2020 to December 2023 were included in the study. According to the 2022 International Globe and Adnexal Trauma Epidemiology Study groups consensus, zone Ⅲ was further divided into Ⅲa zone (5-8 mm posterior to the limbus) and Ⅲb zone (>8 mm posterior to the limbus), with 58 eyes (31%, 58/187) in group Ⅲa and 129 eyes (69%, 129/187) in group Ⅲb. Best corrected visual acuity (BCVA) was examined using the international standard decimal visual acuity chart, converted into the logarithm of the minimum angle of resolution (logMAR) visual acuity when recorded. The injured zone, initial visual acuity, final visual acuity, retinal detachment (RD), uveal prolapse, and proliferative vitreoretinopathy (PVR) were collected. The follow-up time after surgery ≥ 6 months. The final visual acuity and anatomical prognosis of the two groups were observed. Silicone oil dependence, phthisis, and enucleation were defined as poor anatomical outcomes. Multiple linear regression analysis was performed to analyze the impact of zone Ⅲb of OGI on the final visual acuity.Results:At the 6-month follow-up, the logMAR BCVA of group Ⅲa and group Ⅲb was 1.49±1.0 and 2.51±0.85; there was a statistically significant difference in the logMAR BCVA between the two groups ( t=-2.736, P<0.05). Compared with group Ⅲa, the proportion with light perception in group Ⅲb was higher, and the proportions with visual acuity of hand movement, counting fingers, and >0.01 were lower, and the differences were all statistically significant ( P<0.05). Compared with group Ⅲa, RD and PVR were more likely to occur in group Ⅲb, and the differences were all statistically significant ( χ2= 16.696, 8.697; P<0.05). Among the affected eyes in group Ⅲa and group Ⅲb, there were 14 eyes (24.1%, 14/58) and 95 eyes (73.6%, 95/129) with poor final anatomical outcomes respectively; the incidence of poor final anatomical outcomes in group Ⅲb was higher, and the difference was statistically significant ( χ2= 40.332, P<0.01). The results of multiple linear regression analysis showed that initial visual acuity, RD, and uveal prolapse were independent risk factors affecting the final visual acuity (odds ratio=2.407, 4.162, 3.413; P<0.05). Conclusions:Patients with OGI in zone Ⅲb have a worse visual prognosis and a higher incidence of poor anatomical outcomes. The subclassification of zone Ⅲ is helpful for better predicting the prognosis of OGI clinically.
8.Clinical guideline for diagnosis and treatment of nonunion of osteoporotic vertebral fractures (version 2025)
Haipeng SI ; Le LI ; Junjie NIU ; Wencan ZHANG ; Fuxin WEI ; Jinqiu YUAN ; Qiang YANG ; Hongli WANG ; Guangchao WANG ; Shihong CHEN ; Yunzhen CHEN ; Xiaoguang CHENG ; Jianwen DONG ; Shiqing FENG ; Rui GU ; Yong HAI ; Tianyong HOU ; Bo HUANG ; Xiaobing JIANG ; Lei ZANG ; Chunhai LI ; Nianhu LI ; Hua LIN ; Hongjian LIU ; Peng LIU ; Xinyu LIU ; Sheng LU ; Shibao LU ; Chunshan LUO ; Lvy CHAOLIANG ; Lvy WEIJIA ; Xuexiao MA ; Wei MEI ; Chunyang MENG ; Cailiang SHEN ; Chunli SONG ; Ruoxian SONG ; Jiacan SU ; Honglin TENG ; Hui SHENG ; Beiyu WANG ; Bingwu WANG ; Liang WANG ; Xiangyang WANG ; Nan WU ; Guohua XU ; Yayi XIA ; Jin XU ; Youjia XU ; Jianzhong XU ; Cao YANG ; Maowei YANG ; Zibin YANG ; Xiaojian YE ; Hailong YU ; Xijie YU ; Hua YUE ; Zhili ZENG ; Xinli ZHAN ; Hui ZHANG ; Peixun ZHANG ; Wei ZHANG ; Zhenlin ZHANG ; Jianguo ZHANG ; Tengyue ZHU ; Qiang LIU ; Huilin YANG
Chinese Journal of Trauma 2025;41(10):932-945
Nonunion of osteoporotic vertebral fractures (OVF), predominantly affecting the elderly, can lead to intractable pain, vertebral collapse, progressive kyphotic deformity, and neurological impairment, significantly compromising patients′ quality of life. There exists considerable debate on diagnosis and management of OVF, encompassing key issues such as clinical diagnosis and staging criteria for nonunion, surgical indications and procedure selection, and postoperative rehabilitation planning. Currently, there lacks standardized clinical guideline and expert consensus on the diagnosis and management of OVF nonunion in China. To address this gap, Minimally Invasive Surgery Group of Chinese Orthopedic Association, Osteoporosis Committee of Chinese Association of Orthopedic Surgeons, Prevention and Rehabilitation Committee for Osteoporosis of Chinese Association of Rehabilitation Medicine and Minimally Invasive Orthopedic Surgery Branch of China Association for Geriatric Care jointly organized domestic experts in spinal surgery, endocrinology, and rehabilitation to formulate the Clinical guideline for the diagnosis and treatment for nonunion of osteoporotic vertebral fractures ( version 2025), based on existing literature and clinical experience and adhering to principles of scientific rigor and practicality. The guideline provided 13 evidence-based recommendations encompassing diagnosis and treatment of OVF nonunion, aiming to standardize its clinical management.
9.Automated fiber quantification for analyzing relationships of abnormal white matter microstructures and executive dysfunction in children with attention deficit hyperactivity disorder
Fan TAN ; Rui HU ; Yong WU ; Nan HUANG ; Ling SANG ; Wen CHEN
Chinese Journal of Medical Imaging Technology 2025;41(7):1056-1061
Objective To observe the value of automated fiber quantification(AFQ)for analyzing relationships of abnormal white matter microstructures and executive dysfunction in children with attention deficit hyperactivity disorder(ADHD).Methods Brain diffusion tensor imaging(DTI)and white matter structure AFQ analysis were prospectively performed on 83 children with ADHD(ADHD group)and 52 healthy children(healthy control[HC]group).The axial diffusivity(AD),radial diffusivity(RD),mean diffusivity(MD),and fractional anisotropy(FA)of complete fiber bundles and respective segments were obtained and compared between groups.Partial correlation analysis was conducted of fiber bundle diffusion parameters and the scores of Swanson Nolan and Pelham-Ⅳ(SNAP-Ⅳ)and behavior rating inventory of executive function(BRIEF)in ADHD group.Receiver operating characteristic curves were applied to assess the efficacy of each diffusion parameter for evaluating ADHD.Results AD and MD of right anterior thalamic radiation(ATR),and AD of right cingulum bundle(CB)in ADHD group were all higher than those in HC group(FWE corrected,all P<0.000 7).Significant differences of AD of the 54-78 segments of right ATR,MD of the 13-24 and 53-77 segments of right ATR,AD of the 24-26,53-56 and 93-97 segments of right CB,as well as MD of the 72-85 segments of left inferior longitudinal fasciculus(ILF)were found between groups(FWE corrected,all P<0.001).In ADHD group,AD of 54-78 segments of right ATR were negatively correlated with the attention deficit scores,while of 24-26 segments of right CB were negatively correlated with the emotional control scores.MD of right ATR and some segments,AD of right CB and some segments in ADHD group were positively correlated with the scores of task initiation,planning,organization,monitoring and metacognitive index(all P<0.05).The area under the curve(AUC)of segmental fiber bundle diffusion parameter and intact fiber bundle diffusion parameter for evaluating ADHD was 0.881 and 0.808,respectively,the former was higher than the latter(Z=2.435,P<0.05).Conclusion AFQ could be used to analyze relationships of abnormal white matter microstructures and executive dysfunction in ADHD,which could be effectively evaluated based on specific fiber bundle segments.
10.Network pharmacology and molecular docking analysis and animal experimental study of ligustilide regulating H-type blood vessels in prevention and treatment of osteoporosis
Kai WANG ; Hao-nan WEN ; Zhi-jing SONG ; Yong-jia SONG ; Min SONG
Chinese Pharmacological Bulletin 2025;41(3):583-591
Aim To explore the biological mechanism of ligustilide in the prevention and treatment of osteo-porosis by regulating H-type blood vessels,combined with animal experiments for verification,based on net-work pharmacology and molecular docking technology Methods The possible mechanism of ligustilide regu-lating H-type blood vessels to prevent osteoporosis was predicted by network pharmacology.Molecular docking technology was used to verify the binding ability of the core target EGFR to ligustilide.The rat model of osteo-porosis was established and divided into the sham group,model group,ligustilide high,medium and low dose(80,40,20 mg·kg-1)groups.The pathological changes of femur were observed by HE staining.The expressions of CD31,EMCN,OSX+and RUNX2+pro-tein in tibial metaphysis were detected by immunofluo-rescence.The expression of p-EGFR,p-PI3K and p-Akt protein was detected by Western blot.Results The results of network pharmacology showed that a total of 20 intersection targets were obtained.EGFR,PTGS2,ESR1 and ICAM1 were core targets,and mo-lecular docking showed that EGFR had a strong bind-ing ability with ligustilide.The signaling pathways of ligustilide in the prevention and treatment of osteoporo-sis by regulating the expression of H-type blood vessels were mainly enriched in PI3K-Akt,TNF,etc.Com-pared with the model group,ligustilide could signifi-cantly increase the number of trabecular bone and im-prove the destruction of bone microstructure.The ex-pression of CD31,EMCN,OSX+and RUNX2+signifi-cantly increased(P<0.01,P<0.05),the formation of H-type blood vessels were promoted,and the expres-sion of p-EGFR,p-PI3K and p-Akt significantly in-creased(P<0.01,P<0.05).Conclusions Ligusti-lide can increase the expression of H-type blood vessels in bone tissue of osteoporosis model rats,reduce the damage of bone trabecula and improve bone micro-structure effectively.EGFR-mediated PI3K/Akt signa-ling pathway may be the key way to exert its biological effects.

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