1.Mechanism of Buyang Huanwutang in Inhibiting Ferroptosis and Enhancing Neurological Function Recovery After Spinal Cord Injury via GPX4-ACSL4 Axis
Luchun XU ; Guozheng JIANG ; Yukun MA ; Jiawei SONG ; Yushan GAO ; Guanlong WANG ; Jiaojiao FAN ; Yongdong YANG ; Xing YU ; Xiangsheng TANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):20-30
ObjectiveTo explore the mechanism by which Buyang Huanwutang regulates the glutathione peroxidase 4 (GPX4)-acyl-CoA synthetase long-chain family member 4 (ACSL4) axis to inhibit ferroptosis and promote neurological functional recovery after spinal cord injury (SCI). MethodsNinety rats were randomly divided into five groups: sham operation group, model group, low-dose Buyang Huanwutang group (12.5 g·kg-1), high-dose Buyang Huanwutang group (25 g·kg-1), and Buyang Huanwutang + inhibitor group (25 g·kg-1 + 5 g·kg-1 RSL3). The SCI model was established by using the allen method. Tissue was collected on the 7th and 28th days after operation. Motor function was assessed by using the Basso-Beattie-Bresnahan (BBB) scale. Hematoxylin-eosin (HE), Nissl, and Luxol fast blue (LFB) staining were performed to observe spinal cord histopathology. Transmission electron microscopy was used to examine mitochondrial ultrastructure. Immunofluorescence staining was used to detect the number of NeuN-positive cells and the fluorescence intensity of myelin basic protein (MBP), GPX4, and ACSL4. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) was used to analyze the mRNA expression of GPX4 and ACSL4. Enzyme linked immunosorbent assay (ELISA) was performed to measure the levels of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD). Colorimetric assays were used to determine the iron content in spinal cord tissue. ResultsCompared to the sham operation group, the model group exhibited significantly reduced BBB scores (P<0.01), severe pathological damage in spinal cord tissue, and marked mitochondrial ultrastructural disruption. In addition, the model group showed a decrease in the number of NeuN-positive cells (P<0.01), reduced fluorescence intensity of MBP and GPX4 (P<0.01), lower levels of GSH and SOD (P<0.01), and downregulated mRNA expression of GPX4 (P<0.01). Moreover, compared to the sham operation group, the model group had elevated levels of ROS, MDA, and tissue iron content (P<0.01), along with increased fluorescence intensity and mRNA expression of ACSL4 (P<0.01). Compared with the model group and Buyang Huanwutang + inhibitor group, the Buyang Huanwutang group showed significantly improved BBB scores (P<0.05, P<0.01) and exhibited less severe spinal cord tissue damage, reduced edema and inflammatory cell infiltration, increased neuronal survival, and more intact myelin structures. Additionally, mitochondrial ultrastructure was significantly improved in the Buyang Huanwutang group. Compared to the model group and Buyang Huanwutang + inhibitor group, the Buyang Huanwutang group significantly increased the number of NeuN-positive cells and the fluorescence intensity of MBP (P<0.05, P<0.01). Furthermore, Buyang Huanwutang significantly increased the fluorescence intensity and mRNA expression of GPX4 (P<0.01) and decreased the fluorescence intensity and mRNA expression of ACSL4 (P<0.01) compared to the model group and Buyang Huanwutang + inhibitor group. Finally, the Buyang Huanwutang group significantly decreased ROS, MDA, and tissue iron content (P<0.01) and significantly increased GSH and SOD levels (P<0.01) compared to the model group and Buyang Huanwutang + inhibitor group. ConclusionBuyang Huanwutang inhibits ferroptosis through the GPX4/ACSL4 axis, reduces secondary neuronal and myelin injury and oxidative stress, and ultimately promotes the recovery of neurological function.
2.Buyang Huanwu decoction promotes angiogenesis and improves hemorheological parameters after cervical spinal cord injury
Luchun Xu ; Yongdong Yang ; Guozheng Jiang ; Yushan Gao ; Jiawei Song ; Yukun Ma ; Jiaojiao Fan ; Guanlong Wang ; Xing Yu ; Xiangsheng Tang
Journal of Traditional Chinese Medical Sciences 2024;11(4):456-465
Objective:
To explore the effects of Buyang Huanwu decoction (BYHWD) on vascular neogenesis and hemorheological parameters following cervical spinal cord injury (SCI).
Methods:
An acute cervical SCI model was established using 84 female Sprague–Dawley rats. Functional recovery of the rats was evaluated using the forelimb locomotor scale score, forelimb grip strength test, and Basso-Beattie-Bresnahan score. The animals were subsequently euthanized at days 7 and 28 postoperatively. The gross morphology, neuronal survival, and myelin sheath in the injured area were evaluated using hematoxylin and eosin (HE), Nissl, and luxol fast blue (LFB) staining, respectively. Immunofluorescence staining was used to observe CD31 expression 7 days post-injury. Furthermore, the expression of CD31, neuronal nuclear protein (NeuN), and myelin basic protein (MBP) were evaluated 28 days post-injury. Additionally, vascular endothelial growth factor A (VEGFA) and VEGF receptor-2 (VEGFR-2) expression was evaluated using western blotting. Whole-blood viscosity, plasma viscosity, and red blood cell aggregation were measured using a hemorheometer.
Results:
From postoperative days 3–28, motor function in the BYHWD group began to recover considerably compared to the SCI group. BYHWD effectively restored spinal cord histopathology. In addition, the number of NeuN-positive cells, and fluorescence intensity of CD31at 7 and 28 days and MBP significantly increased in the BYHWD group compared with the SCI group (all P < .05). Moreover, this decoction significantly upregulated the expression of VEGFA and VEGFR-2 (all P < .05). BYHWD improved the hemorheology results (i.e., except erythrocyte aggregation index in the low-dose group), revealing statistically significant differences compared with the SCI group (all P < .05).
Conclusion
BYHWD effectively promoted angiogenesis, improved hemorheological parameters, and protected neurons and myelin sheaths, ultimately promoting the recovery of neurological function after cervical SCI in rats. These findings suggest that BYHWD promotes vascular neogenesis through the VEGFA/VEGFR-2 pathway.
3.Effects of Zhu Lian stimulant Type I acupuncture on the expression of brain-derived neurotrophic factor and its receptor TrkB and tissue homogenate cyclic adenosine phosphate in rats with diabetic bladder
Ningjing QIN ; Yushan FAN ; Yujun HE ; Rui LIN ; Hui ZHANG ; Jingwen HUANG
International Journal of Traditional Chinese Medicine 2024;46(1):49-55
Objective:To observe the effects of Zhulian stimulant type Ⅰ acupuncture on the expression of brain-derived neurotrophic factor (BDNF) and its receptor TrkB and tissue homogenate cyclic adenosine phosphate (cAMP) in rats with diabetic bladder (DCP); To explore the mechanism of Zhulian stimulant type Ⅰacupuncture on DCP.Methods:Totally 50 SD rats were divided into control group, model group, Western medicine group, ordinary acupuncture group, Zhulian stimulant type Ⅰ acupuncture treatment group (acupuncture treatment group) according to random number table method, with 10 rats in each group. DCP rat model was prepared by intraperitoneal injection of streptozotocin (STZ), except for the control group. The Western medicine group was given mecobalamine for gavage; acupoints of "Zhongji", "Sanyinjiao", "Liechou" and "Taichong" were selected. The ordinary acupuncture group was treated with ordinary acupuncture technique, and the acupuncture treatment group was treated with Zhulian stimulant type Ⅰ acupuncture, 1 time/d, 30 minutes/time. Samples were taken after 4 weeks of treatment. The maximum bladder volume, residual urine volume and wet weight of the bladder were detected. The morphology of rat bladder was observed by HE staining. The expression level of BDNF was detected by immunohistochemistry. The expression of cAMP was detected by Western blot. The level of TrkB was determined by ELISA. Real-time fluorescence quantitative PCR (RT-PCR) was used to detect the mRNA expressions of BDNF and cAMP.Results:Compared with model group, maximum bladder volume, residual urine volume and wet weight of bladder in Western medicine group, ordinary acupuncture group and acupuncture treatment group decreased ( P<0.01), and those in Western medicine group and acupuncture treatment group were lower than those in ordinary acupuncture group ( P<0.01). The expressions of BDNF mRNA and protein, cAMP mRNA and protein in Western medicine group, ordinary acupuncture group and acupuncture treatment group increased ( P<0.05), and the level of TrkB increased, and the Western medicine group and acupuncture treatment group were higher than that in ordinary acupuncture group ( P<0.05). Conclusions:Zhuliping stimulant type Ⅰ acupuncture has a protective effect on the bladder function of diabetic rats. The mechanism may be related to the up-regulation of BDNF and mRNA, TrkB, cAMP and mRNA expressions.
4.Analysis on Acupoint Selection Rules of Acupuncture in Treating Perimenopausal Syndrome Based on Data Mining
Fangzhi ZHANG ; Yushan FAN ; Furui MIAO ; Yujun HE
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(6):24-31
Objective To explore the acupoint selection rules in the treatment of perimenopausal syndrome(PMS)with acupuncture by using data mining technology.Methods The clinical research literature on acupuncture treatment of PMS was retrieved from CNKI,Wanfang Data,VIP,CBM,Cochrane Library,Embase,PubMed and ScienceDirect from the establishment of the databases to August 16,2023.Excel 2021 was used to establish a prescription database for acupuncture treatment of PMS,and frequency statistics and classification of acupoints were conducted.SPSS Modeler 18.0 and SPSS Statistics 26.0 software were used to conduct association rules,clustering analysis and decision tree analysis,to explore the characteristics and compatibility rules of acupoint selection in acupuncture and moxibustion treatment of PMS.Results Totally 114 articles were included,including 115 acupoint prescriptions,and 108 acupoints with a total frequency of 787.The high-frequency acupoints were Sanyinjiao,Guanyuan,Shenshu,etc.The commonly used meridians were the bladder meridian and Conception Vessel.The majority specific acupoints were intersection acupoints,wu-shu points and back-shu points.The involved acupoints were mostly located in the lower limbs,abdomen and back.Acupuncture techniques often used the method of flat reinforcement and reduction,with the most commonly used waveform of electroacupuncture being the dilatational wave and the duration of needle retention being 10-30 minutes.The decision tree selected three simplified selection paths based on the dependent variable of Guanyuan.The core acupoints group was analyzed,and 14 groups of association rules and 7 effective cluster groups were obtained.Conclusion Acupuncture treatment of PMS follows the therapeutic principles of tonifying the kidney and harmonizing yin and yang.The acupoint combinations and core acupoint groups obtained from data mining can provide reference for clinical and scientific research.
5.Exploration on the Acupoint Selection Law of Acupuncture in Treating Chronic Prostatitis/Chronic Pelvic Pain Syndrome Based on Data Mining
Fangzhi ZHANG ; Yushan FAN ; Furui MIAO ; Yujun HE
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(10):46-53
Objective To explore the clinical acupoint selection law of acupuncture in the treatment of chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS)by data mining.Methods The clinical research literature on acupuncture treatment of CP/CPPS was retrieved from CNKI,VIP,Wanfang Data,China Biology Medicine Database,Embase,PubMed,Cochrane Library and Web of Science from the establishment of the databases to November 17,2023.The prescription database was established using Excel 2021 to perform frequency statistics of acupoints.SPSS Modeler 18.0 and SPSS Statistics 26.0 were used to conduct association rules,factor analysis and clustering analysis,to explore the characteristics and acupoint selection and compatibility law in acupuncture treatment of CP/CPPS.Results A total of 105 articles was included,and 105 acupoint prescriptions were extracted,involving a total of 75 acupoints,with a total frequency of 611 times.The high-frequency acupoints were Zhongji,Sanyinjiao,Guanyuan,etc.The commonly used meridians were Conception Vessel and the bladder meridian.Jiaohui acupoints,Mu acupoints and Wushu acupoints accounted for the vast majority.The involved acupoints were mostly located in the abdomen,lower limbs,and lumbosacral region.The core acupoints was analyzed,and 19 groups of association rules,8 factors and 8 effective cluster groups were obtained.Conclusion Acupuncture treatment of CP/CPPS follows the therapeutic principles of tonifying the kidney,removing dampness,soothing the liver and removing blood stasis.The core acupoints group is Guanyuan-Zhongji-Sanyinjiao,with differentiation of syndromes and compatibility with other acupoints.
6.Discussion on acupoint selection law of acupuncture and moxibustion for postpartum incontinence based on data mining technology
Liangliang HUANG ; Yushan FAN ; Rui LIN ; Sen LUO ; Zhan LI
International Journal of Traditional Chinese Medicine 2024;46(7):932-936
Objective:To explore the acupoint selection law in acupuncture and moxibustion in treating postpartum incontinence by using data mining technology.Methods:The clinical research literature on acupuncture and moxibustion treatment for postpartum incontinence was retrieved from CNKI, Wanfang Data, VIP, CBM, China Medical Journal Full-text Database, PubMed, Embase, Web of Science from the establishment of the databases to February 1, 2023. The acupoint prescription database was established using Excel 2016 and conducting frequency statistics of acupoints, meridians and specific acupoints. Clustering analysis was performed using SPSS Statistics 25.0, and association rule analysis was performed using SPSS Modeler 18.0.Results:A total of 70 articles were included, involving 70 acupuncture prescriptions and including 31 acupoints with a total frequency of 384 times. The top five acupoints were Sanyinjiao (SP6, 46 times), Guanyuan (CV4, 45 times), Zusanli (ST36, 43 times), Zhongji (RN3, 41 times), and Qihai (CV6, 38 times). The commonly used meridians were Conception Vessel, bladder meridian and spleen meridian. The involved acupoints were mostly located in the lower limbs and belly, and most of them were Jiaohui, Mu and Wushu acupoints. Five effective clustering groups were obtained by clustering analysis of high frequency acupoints. The core acupoint combination of association rules was Sanyinjiao (SP6)-Zusanli (ST36)-Guanyuan (CV4)-Qihai (CV6)-Zhongji (RN3).Conclusions:The pathogenesis of postpartum urinary incontinence is mainly characterized by qi deficiency. Acupuncture and moxibustion treatment of postpartum urinary incontinence follows the principles of invigorating qi and tonifying spleen, nourishing lung and tonifying kidney, paying attention to the use of acupoints in abdomen and lower limbs, and using the methods of upper and lower acupoints, anterior and posterior acupoints.
7.Analysis of ankylosing spondylitis animal model based on clinical characteristics of traditional Chinese and Western medicine
Fangzhi ZHANG ; Furui MIAO ; Yushan FAN ; Yujun HE
Chinese Journal of Comparative Medicine 2024;34(8):128-138
Ankylosing spondylitis is an autoimmune disease with sacroiliac arthritis and spinal arthritis as the main manifestations.The disease mainly occurs in young men,has a high disability rate,and is a serious threat to the life and health of patients.Biological agents are expensive,and many adverse reactions to hormones,non-steroidal anti-inflammatory drugs,and anti-rheumatic drugs have been recorded.Traditional Chinese medicine can regulate the immunity and anti-inflammatory effects of the disease,and has good clinical effects.To promote the further study of the pathogenesis of ankylosing spondylitis and the development and screening of therapeutic drugs and therapies,in this paper,we summarize the method and mechanisms of modeling of the existing animal model of ankylosing spondylitis and analyze the advantages and disadvantages of the model.To evaluate the agreements between Chinese and Western medicine clinical characteristics,we compare the characteristics of Chinese and Western medical syndromes of the animal model.Building an animal model of ankylosing spondylitis with a higher degree of consistency between traditional Chinese and Western medicine is the key to innovative research into traditional Chinese medicine method of treating ankylosing spondylitis.To lay the foundation for research into traditional Chinese medicines and acupuncture for ankylosing spondylitis,this paper analyzes the degree of concurrence between the Chinese and Western medicine clinical characteristics of animal models.
8.Inhibiting effects of manual acupuncture on bladder cell apoptosis in rats with diabetic neurogenic bladder
Yujun HE ; Furui MIAO ; Yushan FAN ; Rui LIN ; Ningjing QIN ; Hui ZHANG ; Jingwen HUANG ; Cai HE
Journal of Acupuncture and Tuina Science 2024;22(3):184-194
Objective:To observe the inhibiting effects of manual acupuncture(MA)on bladder cell apoptosis in rats with diabetic neurogenic bladder(DNB)based on the protein and mRNA expression of B-cell lymphoma-leukemia(Bcl)-2,Bcl-2-associated X(Bax)protein,caspase-3,and the protein expression of α-smooth muscle actin(α-SMA),transforming growth factor(TGF)-β in the bladder tissue. Methods:A DNB rat model was established via intraperitoneal injection of streptozotocin(STZ).The rats were randomly divided into a control group,a model group,and an MA group,with 10 rats in each group.For the MA group,MA was applied after modeling.The body mass,fasting blood glucose(FBG),bladder wet weight,and bladder histomorphology were observed.Protein and mRNA expression levels of Bcl-2,Bax,and caspase-3 and the protein expression of α-SMA and TGF-β in the bladder tissue were determined.The apoptotic index of bladder cells was also evaluated. Results:After STZ injection,compared with the control group,the model group and the MA group both showed higher FBG from week 3 and lower body mass from week 9(P<0.05),and had a larger bladder wet weight(P<0.05).Compared with the model group,the MA group showed a smaller bladder wet weight(P<0.05).The histopathological evaluation indicated that MA improved muscle fiber alignment and detrusor cell compensatory hypertrophy in the bladder tissue.In addition,compared with the control group,the apoptotic index increased significantly in the model group and the MA group(P<0.05);the protein and mRNA expression levels of Bax and caspase-3 and the protein expression level of TGF-β in the bladder tissue in the model group and the MA group increased significantly(P<0.05),while the protein and mRNA expression levels of Bcl-2 and the protein expression level of α-SMA in the bladder tissue decreased significantly(P<0.05).Compared with the model group,the apoptotic index of the MA group decreased significantly(P<0.05);the protein and mRNA expression levels of Bax and caspase-3 and the protein expression level of TGF-β in the bladder tissue decreased significantly(P<0.05),while the protein and mRNA expression levels of Bcl-2 and the protein expression level of α-SMA in the bladder tissue increased significantly(P<0.05). Conclusion:MA can protect the bladder by inhibiting the excessive apoptosis of bladder cells,which may be related to the down-regulation of Bax and caspase-3 proteins and mRNAs and TGF-β protein expression,and the up-regulation of Bcl-2 protein and mRNA and α-SMA protein expression.
9.Correlations of serum eukaryotic translation promoter 2α and activated transcription factor 4 levels with renal tissue injury and renal function in diabetic nephropathy patients
Caifeng MAO ; Namin WEI ; Taibin WANG ; Yushan QIAO ; Ruijie FAN ; Zefa LI
Journal of Clinical Medicine in Practice 2024;28(10):73-77
Objective To investigate the correlations of serum eukaryotic translation promoter 2α (eIF2α) and activated transcription factor 4 (ATF4) levels with the degree of renal tissue injury and renal function in patients with diabetic nephropathy (DN). Methods A total of 102 patients with DN (DN group) and 102 patients with simple diabetes (control group) were selected. According to the severe degree of renal tissue damage, the patients with DN were divided into microalbuminuria group (MG group, 35 cases) and dominant albuminuria group (PG group, 41 cases) and renal dysfunction group (RIG group, 26 cases). Serum levels of eIF2α, ATF4, urea nitrogen (BUN), creatinine (Scr), cystatin C (CysC) and estimated glomerular filtration rate (eGFR) were measured. Pearson correlation analysis was used to explore the correlations of eIF2α and ATF4 with BUN, Scr, CysC and eGFR; multivariate Logistic regression analysis was used to explore the risk factors of DN; receiver operating characteristic (ROC) curve was used to analyze the value of eIF2α and ATF4 in diagnostic of DN. Results Serum levels of eIF2α, ATF4, BUN, Scr and CysC in the DN group were higher than those in control group, eGFR was lower than that in control group (
10. Follow-up of people living with HIV/AIDS by primary health care institutions in rural area of Jiangxi province
Pengfei FAN ; Qing YANG ; Yurong MAO ; Qiang HU ; Houlin TANG ; Jian LI ; Yaling LUO ; Fen WANG ; Huanqing ZHAN ; Siming ZANG
Chinese Journal of Epidemiology 2019;40(3):346-349
Objective:
To understand the current status of follow up of people living with HIV/AIDS by health service at grass root in rural area of Jiangxi province and related factors, and provide references for the promotion of the follow up by grass root health service.
Methods:
People living HIV/AIDS aged ≥18 years and diagnosed before 31 December 2017 in 6 townships of Xinjian, Yushan counties and Guixi city were included in the study in Jiangxi province. They had been followed up for more than one time after the first epidemiologic survey. The information about their demographic characteristics and HIV infection status were collected by using self-designed questionnaire. Univariate and multivariate logistic regression analyses were conducted to identify the factors that influencing the acceptance of follow up by grass root health service.
Results:
Of the 373 surveyed HIV infected subjects aged (53.06±16.15) years, 261 were males (70.0


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