1.Establishment and evaluation of a mouse model of acute exacerbation of interstitial pulmonary fibrosis
Wenjing WU ; Siyuan ZHU ; Xingjie SANG ; Qin ZHANG ; Peng ZHAO ; Hailong ZHANG ; Yunping BAI
Chinese Journal of Pathophysiology 2025;41(4):816-824
AIM:A mouse model of acute exacerbation of idiopathic pulmonary fibrosis(AE-IPF)was estab-lished.METHODS:One hundred and twenty male C57BL/6 mice were randomly divided into a negative control group,an IPF group,and an acute exacerbation of interstitial fibrosis(AE-IPF)group.The IPF group received a low dose(3 mg/kg)of bleomycin(BLM)by endotracheal drip on days 0,14,and 28.The AE-IPF group received a high dose(5 mg/kg)of BLM by endotracheal drip on day 56.The control group received an equal volume of saline at different time points.The AE-IPF group was injected with a high dose(5 mg/kg)of BLM via tracheal drip on day 56 on top of the initial IPF induction,while the control group received equal amounts of saline at different time points.Experiments were con-ducted on the 57th,59th,63rd,and 70th days after the initial modeling.Mice were observed for general conditions,CT imaging changes,HE,and Masson staining to assess the degree of alveolitis and fibrosis in lung tissues.Lung function,hydroxyproline(HYP)content in lung tissues,and interleukin-6(IL-6)content in bronchoalveolar lavage fluid(BALF)were also measured.RESULTS:Mice in the AE-IPF group exhibited wheezing,shortness of breath,dyspnea,and weight loss.CT imaging revealed that IPF group mice showed patchy,subpleural reticular fuzzy shadows with irregular thickening of interlobular septa and intralobular linear shadows,along with tractional bronchiectasis.In the AE-IPF group,new ground-glass shadows and solid shadows appeared in addition to the IPF features.AE-IPF group mice demon-strated decreased lung function,elevated lung index,and acute pulmonary edema.HE and Masson staining of AE-IPF group mice showed consistent pathological manifestations of AE-IPF.HYP content in lung tissues,total cell count in BALF,and IL-6 concentration were significantly higher in the AE-IPF group compared to the control group(P<0.05).CONCLUSION:The use of multiple tracheal drip administrations of bleomycin successfully established an AE-IPF ani-mal model in mice.The 63rd day of the experiment was identified as the optimal observation point,as it exhibited the most significant pathological features and clinical symptoms.This model provides ideal conditions for studying AE-IPF patho-genesis and evaluating therapeutic efficacy.
2.Research on The Construction and Application of Multiple Fluorescence Amplification System for Three Kinds of Stains
Yi-Fan BAI ; He-Miao ZHAO ; Jing CHEN ; Hong-Di LIU ; Rui-Qin YANG ; Chong WANG
Progress in Biochemistry and Biophysics 2025;52(4):982-994
ObjectiveA multiplex amplification system was constructed based on the capillary electrophoresis platform for simultaneous detection of saliva, semen, and vaginal secretions using tissue-specific RNA markers. The aim of this study is to identify the tissue origin of suspicious body fluid stains found at crime scenes and determine whether the body fluid stains at the crime scene are one or several types among saliva, semen, and vaginal secretions. MethodsThirty saliva samples, forty semen samples, and forty vaginal secretion samples (half from 2015 and half from 2024) were collected from healthy adult volunteers. Through primer designing, system formulation, and PCR condition optimization, a multiplex fluorescent amplification system was constructed. The specificity, sensitivity, and detection ability for mixed samples of this system were investigated, and it was tested using real crime scene materials. In the primer design stage, to reduce the requirements for RNA template quality, the amplification products were set within 80-300 bp. In the system formulation stage, dominant and subordinate primers were mainly considered. By reducing the concentration of dominant primers and increasing that of subordinate primers, a capillary electrophoresis spectrum with an appropriate peak height ratio was finally obtained. Additionally, gradient experiments were designed to adjust the concentrations of PCR reagents and PCR amplification conditions, and multiple versions of DNA amplification enzymes were optimized to achieve the best experimental results. ResultsThrough statistical analysis, there was no significant difference in the capillary electrophoresis of the 3 types of body fluid samples from the two years (2015 and 2024), demonstrating that the sample preservation method in this study can preserve samples for a relatively long time. The composite amplification system constructed in this study exhibited high specificity for all 3 types of body fluid, with no cross-reactions between the markers of each type of body fluid. The minimum detection thresholds for the 3 types of body fluid reached 0.002 9, 0.001 5, and 0.42 mg/L, respectively. This system also had a high degree of discrimination for mixed samples, especially for semen-saliva mixtures, where each body fluid marker could still be successfully detected when the concentration ratio of semen to saliva was 100:1. Meanwhile, in the two actual cases presented in this article, the application of this composite amplification system performed outstandingly. ConclusionThe composite amplification detection system constructed in this study can achieve the correct screening of saliva, semen, and vaginal secretions, overcoming the problems such as low specificity and sensitivity of marker tests and unbalanced RFU values of each marker in previous studies. The specificity and sensitivity meet the practical work requirements, and the operation is simple. It provides an analytical and identification method for body fluid stains in actual case and is applicable to the identification of the tissue origin of biological evidence at crime scenes involving sexual assault, indecent assault, and other criminal acts. In the future, more types of body fluid markers will be screened to expand the types of body fluids detected by the system, and body fluid-specific cSNP and cInDel genetic markers will be introduced to infer the sources (individuals and types) of mixed and complex stains more accurately.
3.Integrated evidence chain-based effectiveness evaluation of traditional Chinese medicines (Eff-iEC): A demonstration study.
Ye LUO ; Xu ZHAO ; Ruilin WANG ; Xiaoyan ZHAN ; Tianyi ZHANG ; Tingting HE ; Jing JING ; Jianyu LI ; Fengyi LI ; Ping ZHANG ; Junling CAO ; Jinfa TANG ; Zhijie MA ; Tingming SHEN ; Shuanglin QIN ; Ming YANG ; Jun ZHAO ; Zhaofang BAI ; Jiabo WANG ; Aiguo DAI ; Xiangmei CHEN ; Xiaohe XIAO
Acta Pharmaceutica Sinica B 2025;15(2):909-918
Addressing the enduring challenge of evaluating traditional Chinese medicines (TCMs), the integrated evidence chain-based effectiveness evaluation of TCMs (Eff-iEC) has emerged. This paper explored its capacity through a demonstration study that evaluated the effectiveness evidence of six commonly used anti-hepatic fibrosis Chinese patent medicines (CPMs), including Biejiajian Pill (BP), Dahuang Zhechong Pill (DZP), Biejia Ruangan Compound (BRC), Fuzheng Huayu Capsule (FHC), Anluo Huaxian Pill (AHP), and Heluo Shugan Capsule (HSC), using both Eff-iEC and the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) system. The recognition of these CPMs within the TCM academic community was also assessed through their inclusion in relevant medical documents. Results showed that the evidence of BRC and FHC received higher assessments in both Eff-iEC and GRADE system, while the assessments for others varied. Analysis of community recognition revealed that Eff-iEC more accurately reflects the clinical value of these CPMs, exhibiting superior evaluative capabilities. By breaking through the conventional pattern of TCMs effectiveness evaluation, Eff-iEC offers a novel epistemology that better aligns with the clinical realities and reasoning of TCMs, providing a coherent methodology for clinical decision-making, new drug evaluations, and health policy formulation.
4.Ultrasound-guided percutaneous catheterization and drainage combined with polidocanol sclerosis therapy in treatment of thyroid cysts
Anyang LIU ; Yizhou BAI ; Qi QIN ; Xuewei WANG ; Peiliang ZHAO ; Jinyi TIAN ; Dongfang HUO ; Bin LUO
Chinese Journal of General Surgery 2025;40(10):802-805
Objective:To investigate the therapeutic effectiveness of ultrasound-guided percutaneous catheterization for continuous negative pressure drainage combined with polidocanol in treating large thyroid cysts.Method:Clinical data of 38 patients with large thyroid cysts who were treated consecutively with catheter drainage combined with polidocanol sclerotherapy by the same doctor at Beijing Tsinghua Changgung Hospital from Jan 2021 to May 2024 were retrospectively analyzed. The effectiveness and safety were statistically evaluated, and the relationship between drainage volume and cyst volume was analyzed.Results:Among the 38 patients with thyroid cysts who completed the treatment, the median follow-up was 9 months (range: 3-24 months). The effectiveness rate was 92% (35/38), of which 32 cases (84%) met the cure standard. The maximum diameter of the cysts before treatment was (4.8±1.0) cm, and the maximum diameter of the residual nodules after treatment was (1.5±1.1) cm, the difference was statistically significant ( t=17.389, P<0.01). The amount of drainage exudate is related to the volume of the cyst and the maximum diameter before treatment ( t=-3.149, P=0.003; t=-3.057, P<0.005). 19% of patients showed transient low fever after the injection of polidocanol, with no other complications. Conclusion:For large thyroid cysts, ultrasound-guided percutaneous catheterization for continuous negative pressure drainage combined with polidocanol sclerotherapy is a safe and effective method.
5.Application of Forensic Transcriptomics in the Identification of Tissue Origin of Body Fluid Stains
Yi-Fan BAI ; He-Miao ZHAO ; Jing CHEN ; Hong-Di LIU ; Rui-Qin YANG ; Chong WANG
Journal of Forensic Medicine 2025;41(3):260-266
The inference of tissue origin of body fluid stains is crucial for case investigation and court proceedings.However,traditional methods for identification of body fluid stains,such as morpho-logical,chemical,and immunoassay identifications have certain limitations,and there is an urgent need for more efficient methods for confirmatory experiments.In recent years,the rapid development of tran-scriptomics technology has provided new means for the identification of tissue origin of body fluid stains.Different types of RNA in the transcriptome have their own advantages.This paper elaborates in detail on the application of different types of RNA,such as mRNA,miRNA,circRNA,lncRNA,piRNA and microbial transcriptomics in body fluid identification,and summarizes their respective ad-vantages and limitations,in order to provide a reference for related research.
6.Establishment and evaluation of a mouse model of acute exacerbation of interstitial pulmonary fibrosis
Wenjing WU ; Siyuan ZHU ; Xingjie SANG ; Qin ZHANG ; Peng ZHAO ; Hailong ZHANG ; Yunping BAI
Chinese Journal of Pathophysiology 2025;41(4):816-824
AIM:A mouse model of acute exacerbation of idiopathic pulmonary fibrosis(AE-IPF)was estab-lished.METHODS:One hundred and twenty male C57BL/6 mice were randomly divided into a negative control group,an IPF group,and an acute exacerbation of interstitial fibrosis(AE-IPF)group.The IPF group received a low dose(3 mg/kg)of bleomycin(BLM)by endotracheal drip on days 0,14,and 28.The AE-IPF group received a high dose(5 mg/kg)of BLM by endotracheal drip on day 56.The control group received an equal volume of saline at different time points.The AE-IPF group was injected with a high dose(5 mg/kg)of BLM via tracheal drip on day 56 on top of the initial IPF induction,while the control group received equal amounts of saline at different time points.Experiments were con-ducted on the 57th,59th,63rd,and 70th days after the initial modeling.Mice were observed for general conditions,CT imaging changes,HE,and Masson staining to assess the degree of alveolitis and fibrosis in lung tissues.Lung function,hydroxyproline(HYP)content in lung tissues,and interleukin-6(IL-6)content in bronchoalveolar lavage fluid(BALF)were also measured.RESULTS:Mice in the AE-IPF group exhibited wheezing,shortness of breath,dyspnea,and weight loss.CT imaging revealed that IPF group mice showed patchy,subpleural reticular fuzzy shadows with irregular thickening of interlobular septa and intralobular linear shadows,along with tractional bronchiectasis.In the AE-IPF group,new ground-glass shadows and solid shadows appeared in addition to the IPF features.AE-IPF group mice demon-strated decreased lung function,elevated lung index,and acute pulmonary edema.HE and Masson staining of AE-IPF group mice showed consistent pathological manifestations of AE-IPF.HYP content in lung tissues,total cell count in BALF,and IL-6 concentration were significantly higher in the AE-IPF group compared to the control group(P<0.05).CONCLUSION:The use of multiple tracheal drip administrations of bleomycin successfully established an AE-IPF ani-mal model in mice.The 63rd day of the experiment was identified as the optimal observation point,as it exhibited the most significant pathological features and clinical symptoms.This model provides ideal conditions for studying AE-IPF patho-genesis and evaluating therapeutic efficacy.
7.Ultrasound-guided percutaneous catheterization and drainage combined with polidocanol sclerosis therapy in treatment of thyroid cysts
Anyang LIU ; Yizhou BAI ; Qi QIN ; Xuewei WANG ; Peiliang ZHAO ; Jinyi TIAN ; Dongfang HUO ; Bin LUO
Chinese Journal of General Surgery 2025;40(10):802-805
Objective:To investigate the therapeutic effectiveness of ultrasound-guided percutaneous catheterization for continuous negative pressure drainage combined with polidocanol in treating large thyroid cysts.Method:Clinical data of 38 patients with large thyroid cysts who were treated consecutively with catheter drainage combined with polidocanol sclerotherapy by the same doctor at Beijing Tsinghua Changgung Hospital from Jan 2021 to May 2024 were retrospectively analyzed. The effectiveness and safety were statistically evaluated, and the relationship between drainage volume and cyst volume was analyzed.Results:Among the 38 patients with thyroid cysts who completed the treatment, the median follow-up was 9 months (range: 3-24 months). The effectiveness rate was 92% (35/38), of which 32 cases (84%) met the cure standard. The maximum diameter of the cysts before treatment was (4.8±1.0) cm, and the maximum diameter of the residual nodules after treatment was (1.5±1.1) cm, the difference was statistically significant ( t=17.389, P<0.01). The amount of drainage exudate is related to the volume of the cyst and the maximum diameter before treatment ( t=-3.149, P=0.003; t=-3.057, P<0.005). 19% of patients showed transient low fever after the injection of polidocanol, with no other complications. Conclusion:For large thyroid cysts, ultrasound-guided percutaneous catheterization for continuous negative pressure drainage combined with polidocanol sclerotherapy is a safe and effective method.
8.Inhibition of Oxidative Stress of Sciatic Nerve in Diabetic Rats by Tangbikang Granules Regulating AMPK/PGC-1α/SIRT3 Signaling Pathway
Gang LIU ; Yaqi ZHANG ; Lingling QIN ; Chengfei ZHANG ; Qiue ZHANG ; Huizhong BAI ; Yi ZHAO ; Tonghua LIU ; Xiaohong MU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(6):75-82
ObjectiveTo investigate the effect of Tangbikang granules on oxidative stress of sciatic nerve in diabetic rats by regulating adenylate activated protein kinase/peroxisome proliferator-activated receptor γ coactivator-1α/mitochondrial Sirtuins 3 (AMPK/PGC-1α/SIRT3) signaling pathway. MethodThe spontaneous obesity type 2 diabetes model was established using ZDF rats. After modeling, they were randomly divided into high, medium, and low dose Tangbikang granule groups (2.5, 1.25, 0.625 g·kg-1·d-1) and lipoic acid group (0.026 8 g·kg-1·d-1), and the normal group was set up. The rats were administered continuously for 12 weeks after modeling. The blood glucose of rats was detected before intervention and at 4, 8, 12 weeks after intervention. At the 12th week, motor nerve conduction velocity (MNCV), sensory nerve conduction velocity (SNCV), nerve blood flow velocity, mechanical pain threshold, and thermal pain threshold were detected. The sciatic nerve was taken for hematoxylin-eosin (HE) staining to observe the tissue morphology. The ultrastructure of the sciatic nerve was observed by transmission electron microscope. The expression levels of superoxide dismutase (SOD), malondialdehyde (MDA), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in sciatic nerve were determined by enzyme-related immunosorbent assay (ELISA). The mRNA expressions of AMPKα, AMPKβ, PGC-1α, and SIRT3 in sciatic nerve were determined by real-time polymerase chain reaction (Real-time PCR). ResultCompared with the normal group, fasting blood glucose in the model group was increased at each time point (P<0.01). The mechanical pain threshold was decreased (P<0.05), and the incubation time of the hot plate was extended (P<0.01). MNCV, SNCV, and nerve blood flow velocity decreased (P<0.05). The expression level of SOD was decreased (P<0.01). The expression levels of MDA, IL-1β, and TNF-α were increased (P<0.01). The mRNA expression levels of AMPKα, AMPKβ, PGC-1α, and SIRT3 were decreased (P<0.01). The structure of sciatic nerve fibers in the model group was loose, and the arrangement was disordered. The demyelination change was obvious. Compared with the model group, the fasting blood glucose of rats in the high dose Tangbikang granule group was decreased after the intervention of eight weeks and 12 weeks (P<0.01). The mechanical pain threshold increased (P<0.05). The incubation time of the hot plate was shortened (P<0.01). MNCV, SNCV, and Flux increased (P<0.05). The expression level of SOD was increased (P<0.01). The expression levels of MDA, IL-1β, and TNF-α were decreased (P<0.01). The mRNA expression levels of AMPKα, AMPKβ, PGC-1α, and SIRT3 were increased (P<0.01). The sciatic nerve fibers in the high-dose Tangbikang granule group were tighter and more neatly arranged, with only a few demyelinating changes. The high, medium, and low dose Tangbikang granule groups showed a significant dose-effect trend. ConclusionTangbikang granules may improve sciatic nerve function in diabetic rats by regulating AMPK/PGC-1α/SIRT3 signaling pathway partly to inhibit oxidative stress.
9.Electroacupuncture Combined with Point Bloodletting and Cupping for Idiopathic Facial Palsy in Acute Stage of 40 Cases:A Randomized Controlled Trial
Yi'nan QIN ; Lihong YANG ; Yang BAI ; Tianyu XU ; Nana ZHAO ; Zhimei LI ; Yuanhao DU
Journal of Traditional Chinese Medicine 2024;65(14):1458-1463
ObjectiveTo explore the influence of electroacupuncture combined with point bloodletting and cupping for facial nerve function recovery in acute stage of idiopathic facial palsy (IFP). MethodsEighty patients with IFP in the acute stage were randomly divided into 40 cases each in the treatment group and the control group. In the control group, oral prednisone acetate tablets were administered during the acute stage when the disease duration was less than 10 days; and electroacupuncture and flash cupping were provided during the recovery stage when the disease duration was more than 10 days, five times a week. For treatment group in acute stage, the stellate ganglion, vagus nerve stimulation point in the auricular cavity, Yifeng (TE 17) and Tinghui (GB 2) were needled on the affected side on the basis of the treatment of control group, with Yifeng and Tinghui connecting to electroacupuncture apparatus, once a day; point bloodletting and then cupping in Yifeng 2 times a week; in recovery stage, the treatment was the same as that of the control group. Both groups were treated until the 45th day from onset. The primary outcome was the Toronto facial grading system (SFGS), and the secondary outcomes included house-brackmann (H-B) grade, facial disability index (FDI) score, and number of H-B grade-Ⅰ cases. Adverse events were recorded in both groups. ResultsThe SFGS scores of the patients in both groups were higher on the 10th, 30th and 45th days after onset of disease compared with those before the treatment (P<0.05); the H-B grade was lower on the 30th and 45th days after the onset of the disease compared with those before the treatment (P<0.05); and the facial disability index physical function (FDIP) and facial disability index social function (FDIS) scores were higher on the 30th and 45th days after onset of disease (P<0.05). SFGS scores of patients in the treatment group were significantly higher than those of the control group on the 30th and 45th days after onset (P<0.05); H-B grade was significantly lower than that of the control group on the 30th and 45th days after onset (P<0.05); and FDIP scores on the 45th day after onset, and FDIS scores on the 30th and 45th days after onset were significantly higher than those of the control group (P<0.05). At the end of treatment, 77.50% (31 cases) achieved H-B grade-Ⅰ in the treatment group, which was more than 55.00% (22 cases) in the control group (P<0.05). No adverse events occurred in either group. ConclusionElectroacupuncture combined with point bloodletting and cupping for IFP in acute stage can improve the recovery degree of facial nerve function, improve effectiveness, and show a high degree of safety.
10.Construction and Evaluation of Theranostic Near-infrared Fluorescent Probe for Targeting Inflammatory Brain Edema
Jing QIN ; Yong ZHAO ; Caiqin ZHANG ; Bing BAI ; Changhong SHI
Laboratory Animal and Comparative Medicine 2024;44(3):243-250
Objective A novel compound based on near-infrared fluorescence(NIRF)probe was prepared to achieve dynamic monitoring of an inflammatory brain edema model in mice and real-time evaluation of therapeutic effects through in vivo imaging.Methods The NIRF probe IR-783 was chemically linked with clinical brain edema therapeutic drug furosemide(FSM)to obtain the new compound,IR-783-FSM.The ultraviolet fluorescence properties of the compound were evaluated using an ultraviolet spectrophotometer.The uptake of the compound by mouse macrophage cells RAW 264.7 was detected with in vitro cellular experiments.Its cytotoxicity was evaluated through CCK8 assays.A brain edema model was established in BALB/c mice via intraperitoneal injection of lipopolysaccharide(LPS),confirmed by HE staining and dry-wet weight methods for brain tissues.The mice in the brain edema model were divided into control group,IR-783,and IR-783-FSM treatment groups,receiving intraperitoneal injections of PBS,IR-783,and IR-783-FSM,respectively.Real-time in vivo fluorescence imaging was then performed.The mice in each group were euthanized after 10 hours.Ex vivo brain imaging and dry-wet weight measurements were performed to observe the NIRF imaging characteristics and therapeutic effects of IR-783-FSM on brain edema model.Results The newly synthesized compound,IR-783-FSM,retained the excellent near-infrared fluorescence characteristics of IR-783.It could target mouse macrophages with an IC50 of 48.82 μmol/L.A brain edema model could be successfully constructed with intraperitoneal injection of LPS,with significantly higher brain tissue water content compared to the control group(P<0.01).In vivo imaging showed that IR-783-FSM had a significantly stronger fluorescence signal in the brain edema model than IR-783.Compared to the control group,the brain water content was significantly reduced in the 2,5,and 8 mmol/L IR-783-FSM treatment groups(P<0.01).Conclusion The newly synthesized NIRF probe IR-783-FSM facilitates dynamic monitoring of brain edema and real-time evaluation of therapeutic effects.

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