1.Expert Consensus on Clinical Diseases Responding Specifically to Traditional Chinese Medicine: Threatened Abortion
Xinchun YANG ; Shuyu WANG ; Huilan DU ; Songping LUO ; Zhe JIN ; Rong LI ; Xiangyan RUAN ; Qin ZHANG ; Xiaoling FENG ; Shicai CHEN ; Fengjie HE ; Shaobin WEI ; Qun LU ; Yanqin WANG ; Yang LIU ; Qingwei MENG ; Zengping HAO ; Ying LI ; Mei MO ; Xiaoxiao ZHANG ; Ruihua ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(7):241-246
		                        		
		                        			
		                        			Threatened abortion is a common disease of obstetrics and gynecology and one of the diseases responding specifically to traditional Chinese medicine (TCM). The China Association of Chinese Medicine organized experts in TCM obstetrics and gynecology, Western medicine obstetrics and gynecology, and pharmacology to deeply discuss the advantages of TCM and integrated Chinese and Western medicine treatment as well as the medication plans for threatened abortion. After discussion, the experts concluded that chromosome, endocrine, and immune abnormalities were the key factors for the occurrence of threatened abortion, and the Qi and blood disorders in thoroughfare and conception vessels were the core pathogenesis. In the treatment of threatened abortion, TCM has advantages in preventing miscarriages, alleviating clinical symptoms and TCM syndromes, relieving anxiety, regulating reproductive endocrine and immune abnormalities, personalized and diversified treatment, enhancing efficiency and reducing toxicity, and preventing the disease before occurrence. The difficulty in diagnosis and treatment of threatened abortion with traditional Chinese and Western medicine lies in identifying the predictors of abortion caused by maternal factors and the treatment of thrombophilia. Recurrent abortion is the breakthrough point of treatment with integrated traditional Chinese and Western medicine. It is urgent to carry out high-quality evidence-based medicine research in the future to improve the modern diagnosis and treatment of threatened abortion with TCM. 
		                        		
		                        		
		                        		
		                        	
2.Network meta-analysis of the modeling effects of different factors on rabbit models of steroid-induced osteonecrosis of femoral head
Zhixing HU ; Qun LI ; Chao YANG ; Xiaoxiao WANG ; Luochangting FANG ; Wuqiong HOU ; Na LIN ; Weiheng CHEN ; Chunfang LIU ; Ya LIN
Chinese Journal of Tissue Engineering Research 2024;28(6):976-984
		                        		
		                        			
		                        			OBJECTIVE:The rabbit model of steroid-induced osteonecrosis of femoral head is the most commonly used animal model of femoral head necrosis.The pathological changes of the femoral head are close to clinical practice,however,the conditions,methods and evaluation standards of animal models reported in and outside China are not uniform,which leads to the low scientific value of animal models and is difficult to popularize.This study aimed to clarify the influence of different mold-making conditions on the establishment of steroid-induced osteonecrosis of femoral head rabbit model and analyze the appropriate conditions for the successful model establishment. METHODS:We searched the CNKI,WanFang,VIP,CBM,WoS,PubMed and EMbsae databases for the literature on the modeling of steroid-induced osteonecrosis of femoral head rabbits up to April 1,2022,completed the screening of the literature according to the inclusion and exclusion criteria and literature quality evaluation,and extracted the outcome index data in the literature.RevMan Stata and ADDIS statistical software were used to conduct a meta-analysis of the included data. RESULTS:(1)A total of 82 articles with 1 366 rabbits were included in the study.The steroid-induced osteonecrosis of femoral head modeling methods were divided into three types:steroid-alone method,steroid combined lipopolysaccharide method and steroid combined serum method.Among these,33 articles used steroid-alone method;20 articles used steroid combined lipopolysaccharide method;29 articles used steroid combined serum method.(2)Meta-analysis results showed that the three modeling methods significantly increased the rate of empty bone lacunae in the femoral head of steroid-induced osteonecrosis of femoral head rabbits(P<0.001),and significantly decreased the ratio of the trabecular bone area in the femoral head of steroid-induced osteonecrosis of femoral head rabbits(P<0.001).The order of empty bone lacunae rate of each modeling method was:steroid combined with lipopolysaccharide method>steroid-alone method>steroid combined with serum method>normal group,and the order of trabecular bone area rate of each modeling method was:normal group>steroid combined with serum method>steroid-alone method>steroid combined with lipopolysaccharide method.(3)The results of subgroup analysis suggested that the rate of empty bone lacunae in the rabbit model induced by steroid alone might be related to the rabbit variety and the type of steroid used for modeling(difference between groups P<0.05),in which the combined effect amount of New Zealand white rabbits was higher than that of Chinese white rabbits(P<0.05)and Japanese white rabbits,and the combined effect amount of dexamethasone was higher than that of other steroids.The rate of empty bone lacunae induced by steroid combined with lipopolysaccharide was related to the administration mode of lipopolysaccharide and the type of steroid(P<0.05),among which the combined effect of methylprednisolone sodium succinate was significantly higher than that of other steroids(P<0.05),and the combined effect of prednisolone was significantly lower than that of other steroids(P<0.05).The combined effect of lipopolysaccharide 100 μg/kg×twice was significantly lower than 10 μg/kg×twice and 50 μg/kg×twice(P<0.05).The rate of empty bone lacunae in the model induced by steroid combined with serum was related to serum dose and steroid type(P<0.05),among which the combined effect amount of dexamethasone sodium phosphate was significantly higher than other steroid types(P<0.05),and the combined effect amount of dexamethasone was significantly lower than other steroid types(P<0.05);the combined effect amount of serum"10 mL/kg+6 mL/kg"combined dose was lower than other serum doses(P<0.05). CONCLUSION:(1)With the rate of empty bone lacunae and the ratio of trabecular bone area as the judgment standard for the successful establishment of the model,the three modeling methods can successfully construct the rabbit steroid-induced osteonecrosis of femoral head model,of which the steroid combined with lipopolysaccharide method is the best.(2)New Zealand white rabbits and dexamethasone are recommended when selecting the steroid-alone method.Methylprednisolone sodium succinate and low-dose lipopolysaccharide are recommended when selecting the steroid combined with lipopolysaccharide method.Dexamethasone sodium phosphate is recommended when selecting the steroid combined with serum modeling method.
		                        		
		                        		
		                        		
		                        	
3.Mesenchymal stem cell-derived exosomes as a new drug carrier for the treatment of spinal cord injury: A review
Lin-Fei CHENG ; Chao-Qun YOU ; Cheng PENG ; Jia-Ji REN ; Kai GUO ; Tie-Long LIU
Chinese Journal of Traumatology 2024;27(3):134-146
		                        		
		                        			
		                        			Spinal cord injury (SCI) is a devastating traumatic disease seriously impairing the quality of life in patients. Expectations to allow the hopeless central nervous system to repair itself after injury are unfeasible. Developing new approaches to regenerate the central nervous system is still the priority. Exosomes derived from mesenchymal stem cells (MSC-Exo) have been proven to robustly quench the inflammatory response or oxidative stress and curb neuronal apoptosis and autophagy following SCI, which are the key processes to rescue damaged spinal cord neurons and restore their functions. Nonetheless, MSC-Exo in SCI received scant attention. In this review, we reviewed our previous work and other studies to summarize the roles of MSC-Exo in SCI and its underlying mechanisms. Furthermore, we also focus on the application of exosomes as drug carrier in SCI. In particular, it combs the advantages of exosomes as a drug carrier for SCI, imaging advantages, drug types, loading methods, etc., which provides the latest progress for exosomes in the treatment of SCI, especially drug carrier.
		                        		
		                        		
		                        		
		                        	
4.Surveillance of bacterial resistance in tertiary hospitals across China:results of CHINET Antimicrobial Resistance Surveillance Program in 2022
Yan GUO ; Fupin HU ; Demei ZHU ; Fu WANG ; Xiaofei JIANG ; Yingchun XU ; Xiaojiang ZHANG ; Fengbo ZHANG ; Ping JI ; Yi XIE ; Yuling XIAO ; Chuanqing WANG ; Pan FU ; Yuanhong XU ; Ying HUANG ; Ziyong SUN ; Zhongju CHEN ; Jingyong SUN ; Qing CHEN ; Yunzhuo CHU ; Sufei TIAN ; Zhidong HU ; Jin LI ; Yunsong YU ; Jie LIN ; Bin SHAN ; Yunmin XU ; Sufang GUO ; Yanyan WANG ; Lianhua WEI ; Keke LI ; Hong ZHANG ; Fen PAN ; Yunjian HU ; Xiaoman AI ; Chao ZHUO ; Danhong SU ; Dawen GUO ; Jinying ZHAO ; Hua YU ; Xiangning HUANG ; Wen'en LIU ; Yanming LI ; Yan JIN ; Chunhong SHAO ; Xuesong XU ; Wei LI ; Shanmei WANG ; Yafei CHU ; Lixia ZHANG ; Juan MA ; Shuping ZHOU ; Yan ZHOU ; Lei ZHU ; Jinhua MENG ; Fang DONG ; Zhiyong LÜ ; Fangfang HU ; Han SHEN ; Wanqing ZHOU ; Wei JIA ; Gang LI ; Jinsong WU ; Yuemei LU ; Jihong LI ; Qian SUN ; Jinju DUAN ; Jianbang KANG ; Xiaobo MA ; Yanqing ZHENG ; Ruyi GUO ; Yan ZHU ; Yunsheng CHEN ; Qing MENG ; Shifu WANG ; Xuefei HU ; Wenhui HUANG ; Juan LI ; Quangui SHI ; Juan YANG ; Abulimiti REZIWAGULI ; Lili HUANG ; Xuejun SHAO ; Xiaoyan REN ; Dong LI ; Qun ZHANG ; Xue CHEN ; Rihai LI ; Jieli XU ; Kaijie GAO ; Lu XU ; Lin LIN ; Zhuo ZHANG ; Jianlong LIU ; Min FU ; Yinghui GUO ; Wenchao ZHANG ; Zengguo WANG ; Kai JIA ; Yun XIA ; Shan SUN ; Huimin YANG ; Yan MIAO ; Mingming ZHOU ; Shihai ZHANG ; Hongjuan LIU ; Nan CHEN ; Chan LI ; Jilu SHEN ; Wanqi MEN ; Peng WANG ; Xiaowei ZHANG ; Yanyan LIU ; Yong AN
Chinese Journal of Infection and Chemotherapy 2024;24(3):277-286
		                        		
		                        			
		                        			Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5%were gram-positive and 70.5%were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3%,76.7%and 77.9%,respectively.Overall,94.0%of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8%of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2%in the isolates from children and 95.7%in the isolates from adults.The resistance rate to carbapenems was lower than 13.1%in most Enterobacterales species except for Klebsiella,21.7%-23.1%of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1%to 13.3%.The prevalence of meropenem-resistant strains decreased from 23.5%in 2019 to 18.0%in 2022 in Pseudomonas aeruginosa,and decreased from 79.0%in 2019 to 72.5%in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.
		                        		
		                        		
		                        		
		                        	
5.Repair Mechanism of Jianpi Huogu Prescription on Vascular Injury in Alcohol-induced Osteonecrosis of Femoral Head Based on VEGF/VEGFR2/PI3K/Akt Signaling Pathway
Changting FANG-LUO ; Qianqian WANG ; Congcong SUN ; Qun LI ; Chao YANG ; Xiaoxiao WANG ; Zhixing HU ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(8):186-194
		                        		
		                        			
		                        			MethodIn the experiment, 46% vol Red Star Erguotou (10 mL·kg·d-1) was used to establish the AONFH rat model, and the intervention effect of JPHGP at different doses (2.5, 5.0, 10.0 g·kg-1) was observed. Jiangusheng pill (JGS, 1.53 g·kg-1) was selected as the positive control. After 8 weeks of administration, the bone histomorphometry of the femoral head was analyzed by Micro-CT imaging, and the area of medullary microvessels in the femoral head was detected by ink perfusion. The pathological change was observed by hematoxylin and eosin (HE) staining. The protein expressions of Platelet endothelial cell adhesion molecule-1 (CD31), VEGF, VEGFR2, PI3K, phosphor-Akt (p-Akt) and phosphatase and Tensin homologue deleted on chromosome 10 (PTEN) in the femoral head were determined by immunohistochemistry and Western blot. ResultCompared with normal group, the model group presented the fracture and thinning of trabeculae in the femoral head, increased empty bone lacunae, and elevated number and diameter of adipocytes (P<0.01). Micro-CT imaging revealed a decrease in bone mineral density (BMD), bone volume fraction (BV/TV), trabecular thickness (Tb.Th) and trabecular number (Tb.N) (P<0.05, P<0.01) while an increase in bone surface-to-volume ratio (BS/BV) and trabecular separation (Tb.Sp) (P<0.01). The results of ink perfusion showed that the area of medullary microvessels in the femoral head was reduced (P<0.01). Compared with model group, JPHGP lowered the empty bone lacunae rate as well as the number and diameter of adipocytes in the femoral head of AONFH rats. Micro-CT imaging indicated that JPHGP low-dose group had elevated BV/TV, Tb.Th and Tb.N (P<0.05, P<0.01) while decreased BS/BV (P<0.01), and there was an upward trend in BMD while a downward trend in Tb.Sp, but without statistical difference. In addition, JPHGP medium- and high-dose groups had a rise in BMD, BV/TV, Tb.Th and Tb.N (P<0.05, P<0.01), a decrease in BS/BV and Tb.Sp (P<0.05, P<0.01) and enlarged area of medullary microvessels in the femoral head (P<0.05, P<0.01). The expressions of CD31, VEGF, VEGFR2, PI3K, p-Akt in the model group were lower than those in the normal group (P<0.01), and after medium and high doses of JPHGP treatment, the expressions of CD31, PI3K and p-Akt in the femoral head of rats were up-regulated (P<0.01) while the protein expression of PTEN was down-regulated (P<0.01). Moreover, JPHGP up-regulated the expressions of VEGF and VEGFR2 (P<0.05, P<0.01). ConclusionJPHGP can repair the vascular injury in AONFH, and its mechanism may be related to the activation of VEGF/VEGFR2/PI3K/Akt signaling pathway. This study provides certain scientific basis and reference for the clinical application of JPHGP. ObjecctiveTo observe the repair effect of Jianpi Huogu prescription (JPHGP) on vascular injury in experimental alcohol-induced osteonecrosis of femoral head (AONFH), and to explore its mechanism based on vascular endothelial growth factor (VEGF)/VEGFR2/phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. 
		                        		
		                        		
		                        		
		                        	
6.Comparative study on pathological characteristics of four different antigen-induced rheumatoid arthritis mouse models
Chao YANG ; Zhi-xing HU ; Shuang-rong GAO ; Ze-ran YAN ; Luo-chang-ting FANG ; Xiao-xiao WANG ; Qun LI ; Qing-wen TAO ; Chun-fang LIU ; Na LIN
Acta Pharmaceutica Sinica 2023;58(6):1586-1595
		                        		
		                        			
		                        			 Rheumatoid arthritis (RA) is an autoimmune disease driven by antigens and mediated by T cells. Collagen II (CII) and fibrinogen (Fib) are the two main antigens in the pathogenesis of RA. The antigen produced after citrulline modification (Cit) is also one of the inducements to induce the body to produce a pathogenic anti-citrulline protein antibody (ACPA). To provide a reference for RA-related research, this study intends to establish an RA animal model by using CII, Cit-CII, Fib, and Cit-Fib antigens, emulsification with complete Freund's adjuvant and immunization with DBA/1 mice, respectively, to compare the pathological characteristics of RA models induced by different antigens from the aspects of pathology, imaging and serum biochemistry. Animal welfare and experimental process are in accordance with the regulations of the Experimental Animal Ethics Committee of the China Academy of Chinese Medical Sciences. The results showed that the CII, Cit-CII, and Cit-Fib induced mice all had symptoms such as joint redness and swelling, and toe deformation and the clinical score and incidence rate were higher than those of the normal group. The CII group had the most serious lesions, with a incidence rate of 100%, and the Cit-CII and Cit-Fib groups had mild symptoms, with a incidence rate of 25% and 37.5%, respectively; pathological and imaging examination results showed that the joints of mice in CII-induced group showed severe synovial inflammation, cartilage and bone destruction, while those in Cit-CII and Cit-Fib group showed only slight inflammatory infiltration, joint cavity stenosis and bone destruction; the results of serum antibody detection showed that CII, Cit-CII and Cit-Fib groups all produced high levels of anti-cyclic citrullinated peptide (CCP) antibodies, among which, Cit-Fib group > Cit-CII group > CII group > Fib group, and both Cit-CII and Cit-Fib groups produced high levels of citrullinated epitope-specific antibodies, while the total IgG level was the highest in CII group; serum ELISA and RT-PCR analysis of joint tissue showed that the expression of pro-inflammatory factors and bone destruction-related molecules increased most significantly in the CII-induced group, followed by Cit-Fib and Cit-CII. The above results showed that among the four different antigens, the symptoms and conditions of arthritis in RA mice induced by CII were the most serious, and IgG instead of anti-CCP antibody was its typical immunological feature, and CII could be the first choice for the model of RA mice; Cit-Fib has certain immunogenicity, can partially induce the symptoms and conditions of RA arthritis in mice, and produce high-level anti-CCP antibody and anti-Cit-Fib antibody, which is more suitable for the study of citrulline-related RA; although Cit-CII has certain immunogenicity, the incidence, and severity of RA arthritis induced by Cit-CII in mice are low. 
		                        		
		                        		
		                        		
		                        	
7.Protective Effect of Jianpi Huogu Prescription on Functional Injury of Vascular Endothelial Cells Caused by Alcohol Based on Akt/JNK/p38 MAPK Signaling Pathway
Xiaoxiao WANG ; Lianhua HE ; Changting FANG-LUO ; Qun LI ; Chao YANG ; Zhixing HU ; Weiheng CHEN ; Chunfang LIU ; Lan HAN ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(11):64-71
		                        		
		                        			
		                        			ObjectiveTo investigate the protective effect of Jianpi Huogu prescription (JPHGP) on the functional injury of vascular endothelial cells caused by alcohol and explore its mechanism based on protein kinase B/c-Jun amino-terminal kinase/p38 MAPK (Akt/JNK/p38 MAPK) signaling pathway. MethodThrough chick embryo allantoic membrane, thoracic aortic ring, and migration, invasion, adhesion, and lumen formation of human umbilical vein endothelial cells (HUVEC), the effect of JPHGP with different concentrations (8, 16 and 32 μg·L-1) on angiogenesis was observed in the presence or absence of alcohol. The expression levels of phosphorylation of Akt, JNK, and p38 MAPK were determined by Western blot. ResultAs compared with the normal group, the number and length of capillaries around the arterial ring in the model group were decreased, and the migration, invasion, and lumen formation capacity of HUVEC were decreased (P<0.05, P<0.01). After treatment with 16 and 32 μg·L-1 JPHGP, the length of neovascularization in chick embryo allantoic membrane was significantly increased (P<0.05, P<0.01). Compared with the model group, the 8, 16, and 32 μg·L-1 JPHGP groups increased the number of capillaries around the thoracic aortic ring in a concentration-dependent manner (P<0.05, P<0.01), and the 32 μg·L-1 JPHGP group increased the length of capillaries around the thoracic aortic ring (P<0.05). The 16 and 32 μg·L-1 JPHGP groups enhanced the migration, invasion, and lumen formation capacity of HUVEC. The results of Western blot showed that, as compared with the normal group, the protein expression levels of p-JNK/JNK, p-p38 MAPK/p38 MAPK, and p-Akt/Akt were significantly decreased in the model group (P<0.01), and as compared with the model group, the protein expression levels of p-p38 MAPK/p38 MAPK and p-Akt/Akt were significantly increased in the 8, 16, and 32 μg·L-1 JPHGP groups (P<0.01) and the protein expression level of p-JNK/JNK was increased significantly in the 16 and 32 μg·L-1 JPHGP groups (P<0.01). ConclusionJPHGP has a protective effect on the functional injury of vascular endothelial cells caused by alcohol, and its mechanism may be related to the activation of Akt/JNK/p38 MAPK signaling pathway. Relevant research results will provide certain scientific basis for clarifying the effect of JPHGP on 'invigorating spleen and promoting blood circulation'. 
		                        		
		                        		
		                        		
		                        	
8.Mechanism of Yanghe Decoction against subcutaneous tumor in pulmonary metastasis from breast cancer through HIF-1α signaling pathway regulating glycolysis:based on network pharmacology and animal experiment.
Yang-Jing LIU ; Xiao-Liu LI ; Chao-Qun MA ; De-Xuan CHEN ; Gao-Yuan WANG ; Tai-Yang ZHU
China Journal of Chinese Materia Medica 2023;48(9):2352-2359
		                        		
		                        			
		                        			This study aims to explore the mechanism of Yanghe Decoction(YHD) against subcutaneous tumor in pulmonary metastasis from breast cancer, which is expected to lay a basis for the treatment of breast carcinoma with YHD. The chemical components of medicinals in YHD, and the targets of the components were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and SwissTargetPrediction. The disease-related targets were searched from GeneCards and Online Mendelian Inheritance in Man(OMIM). Excel was employed to screen the common targets and plot the Venn diagram. The protein-protein interaction network was constructed. R language was used for Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment. A total of 53 female SPF Bablc/6 mice were randomized into normal group(same volume of normal saline, ig), model group(same volume of normal saline, ig), and low-dose and high-dose YHD groups(YHD, ig, 30 days), with 8 mice in normal group and 15 mice in each of the other groups. Body weight and tumor size was measured every day. Curves for body weight variation and growth of tumor in situ were plotted. In the end, the subcutaneous tumor sample was collected and observed based on hematoxylin and eosin(HE) staining. The mRNA and protein levels of hypoxia inducible factor-1α(HIF-1α), pyruvate kinase M2(PKM2), lactate dehydrogenase A(LDHA), and glucose transporter type 1(GLUT1) were detected by PCR and Western blot. A total of 213 active components of YHD and 185 targets against the disease were screened out. The hypothesis that YHD may regulate glycolysis through HIF-1α signaling pathway to intervene in breast cancer was proposed. Animal experiment confirmed that the mRNA and protein levels of HIF-1α, PKM2, LDHA, and GLUT1 in the high-and low-dose YHD groups were lower than those in the model group. YHD has certain inhibitory effect on subcutaneous tumor in pulmonary metastasis from breast cancer in the early stage, which may intervene pulmonary metastasis from breast cancer by regulating glycolysis through HIF-1α signaling pathway.
		                        		
		                        		
		                        		
		                        			Female
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		                        			Mice
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		                        			Animals
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		                        			Glucose Transporter Type 1/genetics*
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		                        			Network Pharmacology
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		                        			Animal Experimentation
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		                        			Saline Solution
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		                        			Drugs, Chinese Herbal/therapeutic use*
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		                        			Medicine, Chinese Traditional
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		                        			Signal Transduction
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		                        			Glycolysis
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		                        			RNA, Messenger
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		                        			Neoplasms/drug therapy*
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		                        			Molecular Docking Simulation
		                        			
		                        		
		                        	
9.Characteristic changes of blood stasis syndrome in rat model of steroid-induced femoral head necrosis based on the combination of disease, syndrome, and symptom.
Zhi-Xing HU ; Chao YANG ; Luo-Chang-Ting FANG ; Xiao-Xiao WANG ; Qun LI ; Wei-Heng CHEN ; Yan-Qiong ZHANG ; Ya LIN ; Chun-Fang LIU ; Na LIN
China Journal of Chinese Materia Medica 2023;48(22):6128-6141
		                        		
		                        			
		                        			The approach combining disease, syndrome, and symptom was employed to investigate the characteristic changes of blood stasis syndrome in a rat model of steroid-induced osteonecrosis of the femoral head(SONFH) during disease onset and progression. Seventy-two male SD rats were randomized into a healthy control group and a model group. The rat model of SONFH was established by injection of lipopolysaccharide(LPS) in the tail vein at a dose of 20 μg·kg~(-1)·d~(-1) on days 1 and 2 and gluteal intramuscular injection of methylprednisolone sodium succinate(MPS) at a dose of 40 mg·kg~(-1)·d~(-1) on days 3-5, while the healthy control group received an equal volume of saline. The mechanical pain test, tongue color RGB technique, gait detection, open field test, and inclined plane test were employed to assess hip pain, tongue color, limping, joint activity, and lower limb strength, respectively, at different time points within 21 weeks of modeling. At weeks 2, 4, 8, 12, 16, and 21 after modeling, histopathological changes of the femoral head were observed by hematoxylin-eosin(HE) staining and micro-CT scanning; four coagulation items were measured by rotational thromboelastometry; and enzyme-linked immunosorbent assay(ELISA) was employed to determine the levels of six blood lipids, vascular endothelial growth factor(VEGF), endothelin-1(ET-1), nitric oxide(NO), tissue-type plasminogen activator(t-PA), plasminogen activator inhibitor factor-1(PAI-1), bone gla protein(BGP), alkaline phosphatase(ALP), receptor activator of nuclear factor-κB(RANKL), osteoprotegerin(OPG), and tartrate-resistant acid phosphatase 5b(TRAP5b) in the serum, as well as the levels of 6-keto-prostaglandin 1α(6-keto-PGF1α) and thromboxane B2(TXB2) in the plasma. The results demonstrated that the pathological alterations in the SONFH rats were severer over time. The bone trabecular area ratio, adipocyte number, empty lacuna rate, bone mineral density(BMD), bone volume/tissue volume(BV/TV), trabecular thickness(Tb.Th), trabecular number(Tb.N), bone surface area/bone volume(BS/BV), and trabecular separation(Tb.Sp) all significantly increased or decreased over the modeling time after week 4. Compared with the healthy control group, the mechanical pain threshold, gait swing speed, stride, standing time, and walking cycle of SONFH rats changed significantly within 21 weeks after modeling, with the greatest difference observed 12 weeks after modeling. The time spent in the central zone, rearing score, and maximum tilt angle in the open field test of SONFH rats also changed significantly over the modeling time. Compared with the healthy control group, the R, G, and B values of the tongue color of the model rats decreased significantly, with the greatest difference observed 11 weeks after modeling. The levels of total cholesterol(TC), total triglycerides(TG), low-density lipoprotein-cholesterol(LDL-C), and apoprotein B(ApoB) in the SONFH rats changed significantly 4 and 8 weeks after modeling. The levels of VEGF, ET-1, NO, t-PA, PAI-1, 6-keto-PGF1α, TXB2, four coagulation items, and TXB2/6-keto-PGF1α ratio in the serum of SONFH rats changed significantly 4-16 weeks after modeling, with the greatest differences observed 12 weeks after modeling. The levels of BGP, TRAP5b, RANKL, OPG, and RANKL/OPG ratio in the serum of SONFH rats changed significantly 8-21 weeks after modeling. During the entire onset and progression of SONFH in rats, the blood stasis syndrome characteristics such as hyperalgesia, tongue color darkening, gait abnormalities, platelet, vascular, and coagulation dysfunctions were observed, which gradually worsened and then gradually alleviated in the disease course(2-21 weeks), with the most notable differences occurred around 12 weeks after modeling.
		                        		
		                        		
		                        		
		                        			Rats
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		                        			Male
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		                        			Animals
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		                        			Femur Head/pathology*
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		                        			Plasminogen Activator Inhibitor 1/adverse effects*
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		                        			Vascular Endothelial Growth Factor A
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		                        			Femur Head Necrosis/pathology*
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		                        			Rats, Sprague-Dawley
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		                        			Steroids
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		                        			Pain
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		                        			Cholesterol
		                        			
		                        		
		                        	
10.Intervention of Osteoking in Rats with Myofascial Pain Syndrome
Xiaoxiao WANG ; Qun LI ; Zhixing HU ; Changting FANGLUO ; Ruirui MING ; Tengteng XU ; Chao YANG ; Wuqiong HOU ; Lili WANG ; Yini JIANG ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(24):19-29
		                        		
		                        			
		                        			ObjectiveTo clarify the intervention effect of Osteoking (OK) in rats with myofascial pain syndrome (MPS) and preliminarily explore the pharmacological mechanism of OK in relieving chronic pain from the perspective of anti-inflammatory disease. MethodThe 60 SD rats were divided into normal group, model group, low, medium, and high dose OK groups (0.66, 1.31, 2.63 mL·kg-1), and positive celecoxib group (21 mg·kg-1). The MPS rat model was established by beating combined with the centrifugal exercise method, and the OK and celecoxib were given at the same time. SMALGO paw pressure pain manometer detected the shock pain point tenderness threshold of rats, and the Von-Frey needle and acetone stimulation method detected the mechanical hyperalgesia threshold and cold hyperalgesia stimulation response respectively. Eight weeks and 10 weeks after modeling, the spontaneous discharge state and convulsion response of MPS rats were determined by electromyograph (EMG) instrument. The gait changes of MPS rats were detected using a CatWalk gait analyzer. The expression levels of interleukin-1 β (IL-1β), tumor necrosis factor-α (TNF-α), substance P (SP), and bradykinin (BK) were measured by enzyme-linked immunosorbent assay (ELISA). The protein expression levels of nuclear transcription factor-κB (NF-κB) inhibiting protein α (IκBα), phosphorylates (p)- IκBα, NF-κB p65, and p-NF-κB p65 were detected in MPS rats by Western blot. The positive expression of p-NF-κB p65 was detected by immunofluorescence. ResultCompared with the normal group, the model group shows 100% positive rates for EMG signal and local convulsions response at both the 8th and 10th weeks. The tenderness threshold and mechanical hyperalgesia threshold are significantly reduced. Cold hyperalgesia score is significantly increased, and gait is abnormal. The expression levels of serum and trigger points IL-1β, TNF-α, SP, BK, p-IκBα, and p-NF-κB p65, as well as the positive expression intensity of p-NF-κB p65 are significantly increased (P<0.01). Compared with the model group, the positive rate of EMG detection and local convulsion response is significantly reduced in the medium and high dose OK groups (P<0.05). The tenderness threshold and mechanical hyperalgesia threshold increase significantly in the medium and high dose OK groups, and the cold hyperalgesia score is significantly reduced in the high dose OK group (P<0.01). The standing time, swing time, and walking period are significantly increased. The swing speed, maximum contact area, and maximum contact intensity are significantly decreased in the high dose OK group (P<0.05). Moreover, the protein expression levels of p-IκBα/IκBα and p-NF-κB p65/NF-κB p65 are significantly reduced in the medium and high dose OK groups (P<0.05,P<0.01). The positive expression intensity of p-NF-κB p65 is significantly decreased in the high dose OK group (P<0.01). ConclusionThe mechanism of OK in relieving the pain in trigger points of MPS and improving gait abnormalities is related to the downregulation of the NF-κB p65 inflammatory signaling pathway to reduce the expression of inflammatory factors and pain mediators in blood and trigger point tissue. 
		                        		
		                        		
		                        		
		                        	
            
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