1.Shaoyaotang Alleviates Damage of Tight Junction Proteins in Caco-2 Cell Model of Inflammation by Regulating RhoA/ROCK Pathway
Nianjia XIE ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Yuting YANG ; Bo ZOU ; Da ZHAO ; Yi LU ; Mingsheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):70-77
		                        		
		                        			
		                        			ObjectiveTo investigate the protective effect and mechanism of Shaoyaotang (SYD) on the lipopolysaccharide (LPS)-induced damage of tight junction proteins in the human colorectal adenocarcinoma (Caco-2) cell model of inflammation via the Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil forming protein kinase (ROCK) pathway. MethodsCaco-2 cells were grouped as follows: Blank, model (LPS, 10 mg·L-1), SYD-containing serum (10%, 15%, and 20%), and inhibitor (Fasudil, 25 μmol·L-1). After 24 hours of intervention, the cell viability in each group was examined by the cell-counting kit 8 (CCK-8) method. Enzyme-linked immunosorbent assay was employed to determine the levels of endothelin-1 (ET-1), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were employed to determine the mRNA and protein levels, respectively, of RhoA, ROCK2, claudin-5, and zonula occludens-1 (ZO-1) in cells of each group. ResultsCompared with the blank group, the model group showcased a marked reduction in the cell viability (P<0.01), elevations in the levels of ET-1, TNF-α, IL-1β, and IL-6 (P<0.01), declines in both mRNA and protein levels of ZO-1 and claudin-5 (P<0.01), and rises in mRNA and protein levels of RhoA and ROCK2 (P<0.01). Compared with the model group, the Shaoyaotang-containing serum (10%, 15%, and 20%) groups had enhanced cell viability (P<0.01), lowered levels of ET-1, TNF-α, IL-1β, and IL-6 (P<0.01), up-regulated mRNA and protein levels of ZO-1 and claudin-5 (P<0.05, P<0.01), and down-regulated mRNA and protein levels of RhoA and ROCK2 (P<0.01). Moreover, the inhibitor group and the 15% and 20% Shaoyaotang-containing serum groups had lower levels of ET-1, TNF-α, IL-1β, and IL-6 (P<0.05, P<0.01), higher mRNA and protein levels of ZO-1 and claudin-5 (P<0.05, P<0.01), and lower mRNA and protein levels of RhoA and ROCK2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can lower the levels of LPS-induced increases in levels of inflammatory cytokines and endothelin to ameliorate the damage of tight junction proteins of the Caco-2 cell model of inflammation by regulating the expression of proteins in the RhoA/ROCK pathway. 
		                        		
		                        		
		                        		
		                        	
2.Shaoyaotang Alleviates Damage of Tight Junction Proteins in Caco-2 Cell Model of Inflammation by Regulating RhoA/ROCK Pathway
Nianjia XIE ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Yuting YANG ; Bo ZOU ; Da ZHAO ; Yi LU ; Mingsheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):70-77
		                        		
		                        			
		                        			ObjectiveTo investigate the protective effect and mechanism of Shaoyaotang (SYD) on the lipopolysaccharide (LPS)-induced damage of tight junction proteins in the human colorectal adenocarcinoma (Caco-2) cell model of inflammation via the Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil forming protein kinase (ROCK) pathway. MethodsCaco-2 cells were grouped as follows: Blank, model (LPS, 10 mg·L-1), SYD-containing serum (10%, 15%, and 20%), and inhibitor (Fasudil, 25 μmol·L-1). After 24 hours of intervention, the cell viability in each group was examined by the cell-counting kit 8 (CCK-8) method. Enzyme-linked immunosorbent assay was employed to determine the levels of endothelin-1 (ET-1), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were employed to determine the mRNA and protein levels, respectively, of RhoA, ROCK2, claudin-5, and zonula occludens-1 (ZO-1) in cells of each group. ResultsCompared with the blank group, the model group showcased a marked reduction in the cell viability (P<0.01), elevations in the levels of ET-1, TNF-α, IL-1β, and IL-6 (P<0.01), declines in both mRNA and protein levels of ZO-1 and claudin-5 (P<0.01), and rises in mRNA and protein levels of RhoA and ROCK2 (P<0.01). Compared with the model group, the Shaoyaotang-containing serum (10%, 15%, and 20%) groups had enhanced cell viability (P<0.01), lowered levels of ET-1, TNF-α, IL-1β, and IL-6 (P<0.01), up-regulated mRNA and protein levels of ZO-1 and claudin-5 (P<0.05, P<0.01), and down-regulated mRNA and protein levels of RhoA and ROCK2 (P<0.01). Moreover, the inhibitor group and the 15% and 20% Shaoyaotang-containing serum groups had lower levels of ET-1, TNF-α, IL-1β, and IL-6 (P<0.05, P<0.01), higher mRNA and protein levels of ZO-1 and claudin-5 (P<0.05, P<0.01), and lower mRNA and protein levels of RhoA and ROCK2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can lower the levels of LPS-induced increases in levels of inflammatory cytokines and endothelin to ameliorate the damage of tight junction proteins of the Caco-2 cell model of inflammation by regulating the expression of proteins in the RhoA/ROCK pathway. 
		                        		
		                        		
		                        		
		                        	
3.Discussion of factors influencing preclinical studies based on fecal bacteria transplantation in mice
Shiqi SUN ; Lu LIU ; Shuangyuan HU ; Yuyan WANG ; Mingsheng SUN ; Ling ZHAO
Acta Laboratorium Animalis Scientia Sinica 2024;32(1):100-117
		                        		
		                        			
		                        			Fecal microbiota transplantation(FMT)is a therapeutic approach that targets intestinal microorganisms by transplanting fecal microorganisms from healthy individuals into the gastrointestinal tract of diseased individuals,thus restoring the recipient's disordered gastrointestinal microbiota by restructuring the intestinal flora.However,the mechanism of action and adverse effects of FMT in different diseases have not yet been clarified,thus limiting its wide clinical application.Its use still relies on in-depth preclinical studies;however,highly inconsistent or incomplete experimental details provided in current reports,coupled with a lack of authoritative standards and recommendations,seriously affect the interpretation of the study findings and replication of the experimental procedures,as well as hindering the clinical translation of the result.We therefore review and discuss the key steps of recipient selection and graft sample collection,storage,graft material preparation,and grafting route,with the aim of improving the utilization of experimental animals,consumables,and labor,and providing method ological recommendations and references to achieve replicability and standardization of preclinical FMT studies.
		                        		
		                        		
		                        		
		                        	
4.A Case Report of Multidisciplinary Management of a Patient with Schimke Immuno-Osseous Dysplasia
Juan DING ; Wei WANG ; Juan XIAO ; Yan ZHANG ; Huijuan ZHU ; Wen ZHANG ; Peng GAO ; Limeng CHEN ; Wei LYU ; Xuan ZOU ; Xiaoyi ZHAO ; Hongmei SONG ; Mingsheng MA
JOURNAL OF RARE DISEASES 2024;3(4):465-470
Schimke immuno-osseous dysplasia (SIOD)caused by 
5.The Study of Anti-inflammatory Effect of Acupuncture in Rats with Chronic Migraine Based on the α7nAChRs
Yuqi ZHENG ; Mingsheng SUN ; Yi LIU ; Mengdi ZHOU ; Leimiao YIN ; Ling ZHAO ; Dingjun CAI
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(4):1050-1058
		                        		
		                        			
		                        			Objective To investigate the effect of acupuncture on the inflammatory changes of α7 nicotinic acetylcholine receptors(α7 nicotinic acetylcholine receptors,α7nAChRs)in rats with chronic migraine(CM).Methods Adult male SD rats were randomly divided into control group(VEH group),model group(NTG group),acupuncture group(TA group),α7nAChRs antagonist group(MLA group)and α7nAChRs agonist group(TA group)according to basic pain threshold.The rat model of CM was established by repeated subcutaneous injection of nitroglycerin(NTG)at the neck and back every other day.TA group and MLA group received acupuncture treatment one hour before NTG injection,20 minutes every day for 9 consecutive days.The MLA group was intraperitoneally injected with α7nAChRs antagonist MLA half an hours before acupuncture,and the PNU group was intraperitoneally injected with α7nAChRs agonist PNU-282987 half an hours before NTG injection,both groups were injected continuously for 9 days.The changes of paw withdrawal mechanical threshold(Paw Withdrawal Mechanical Threshold,PWMT)and tail-flick latency(Tail-Flick Latency,TFL)were detected by Von Frey and thermal radiation pain meter.The contents of inflammatory factors IL-1β,TNF-α and TGF-β in TNC and serum were determined by ELISA.The relative fluorescence intensity and co-expression of GFAP and α7nAChRs in TNC were detected by immunofluorescence dual labeling method.Results Compared with VEH group,PWMT and TFL in NTG group were significantly decreased(P<0.05 or P<0.01),the contents of IL-1β(P<0.01)and TNF-α(P<0.01,P<0.05)in serum and TNC were significantly increased;and in TNC,the astrocytes were activated significantly(P<0.01),while the relative fluorescence intensity of α7nAChRs and the co-expression of GFAP and α7nAChRs were significantly decreased(P<0.05,P<0.01).Compared with NTG group,PWMT and TFL in TA group were increased(P<0.05 or P<0.01),the contents of IL-1β(P<0.01)and TNF-α(P<0.01,P<0.05)in serum and TNC were increased;and in TNC,the relative fluorescence intensity of GFAP was significantly decreased(P<0.01),however the relative fluorescence intensity of α7nAChRs and the co-expression of GFAP and α7nAChRs were significantly increased(P<0.01).Compared with MLA group,PWMT and TFL in TA groups and PNU groups were significantly increased(P<0.05 or P<0.01).The contents of IL-1β(P<0.05)and TNF-α(P<0.01)in serum and TNC of TA group were increased,while the contents of TGF-β(P<0.05)were decreased.The contents of IL-1β in serum and TNC of PNU group were decreased(P<0.01,P<0.05),while the contents of TGF-β(P<0.01)were significantly increased.Conclusions Acupuncture can effectively relieve CM inflammatory response and hypersensitivity to pain,and its anti-inflammatory and analgesic effects may be related to the up-regulation of α7nAChRs expression.
		                        		
		                        		
		                        		
		                        	
6.Microbial Diversity in Rhizosphere Soil of Gastrodia elata with Different Yields
Yingzi LUO ; Mingjin HUANG ; Dachang WANG ; Cheng LI ; Gang GUO ; Hongchang LIU ; Mingsheng ZHANG ; Zhi ZHAO ; Songlin RUAN ; Tingchi WEN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(1):133-140
		                        		
		                        			
		                        			ObjectiveTo analyze the microbial diversity in the rhizosphere soil of Gastrodia elata with different yields and explore the influence of soil microorganisms on the yield of G. elata. MethodThe experiment adopted the 16S DNA and ITS high-throughput sequencing technologies to study the diversity of the bacterial and fungal community in the rhizosphere soil of G. elata with high yield (GC) and low yield (DC). ResultProteobacteria, Firmicutes, and other unidentified Bacteria were dominant in the rhizosphere soil of G. elata. The dominant rhizosphere fungi were Ascomycota, Basidiomycota, and Mortierellomycota. There was no significant difference in microbial community abundance in the high-yield and low-yield rhizosphere soil of G. elata, but there was a significant difference in species composition. Thirty-eight microbes such as Bradyrhizobium, Schleiferilactobacillus, and Archaeorhizomyces were gathered in large numbers in the high-yield rhizosphere soil, and thirty microbes such as Fusarium, Coprinellus, and Nitrosotalea were gathered in large numbers in the low-yield rhizosphere soil. At the level of genus and species, there were six different species in the high-yield and low-yield rhizosphere soil of G. elata, among which Russula mariae, Archeaeorhizomyces, and Ilyonectria were gathered in the high-yield rhizosphere soil of G. elata, while Nitrosotalea, Coprinellus disserminatus, and Fusarium were gathered in the low-yield rhizosphere soil of G. elata. ConclusionThere are different microorganisms in the rhizosphere soil of G. elata with different yields, and it is speculated that these microorganisms are related to the yields of G. elata. The research results are expected to provide a vital theoretical basis for the follow-up study of the high yield of G. elata. 
		                        		
		                        		
		                        		
		                        	
7.Treatment of Allergic Rhinitis with Traditional Chinese Medicine: A Review
Linjing ZHANG ; Yingying ZHAO ; Lili LIU ; Zheng GONG ; Mingsheng ZHANG ; Jianhua LIU ; Zijun DONG ; Jingyi HU ; Xiaoxiao ZHANG ; Zhanfeng YAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(9):268-273
		                        		
		                        			
		                        			This article reviews the clinical studies about the treatment of allergic rhinitis (AR) with traditional Chinese medicine (TCM) in recent years. AR is a common and frequently occurring disease in the department of otolaryngology. The common manifestations of AR include nasal congestion, sneezing, runny nose, and nasal itching. AR, belonging to the category of rhinitis, is a dominant disease in the TCM treatment of otorhinolaryngologic diseases and head and neck diseases. The ancient literature has laid a solid theoretical basis on the etiology and pathogenesis of AR. According to the ancient literature, the theoretical basis, and their own experience, modern doctors classified the causes of AR into the disorders in Zang-fu organs and six meridians. Most of the explanations focus on the disorders in Zang-fu organs, especially the lung, spleen, and kidney. The clinical studies in this field mainly involve TCM treatment alone, integrated TCM and Western medicine treatment, and TCM external treatment. These therapies have good efficacy in the clinical treatment of AR. Among them, TCM treatment alone has significant advantages in alleviating the symptoms and nasal signs, declining the scores of related scales, and reducing the eosinophil count (EOS) of nasal secretions, with definite long-term efficacy. Integrated TCM and Western medicine treatment can complement with each other. The TCM external treatment methods include acupuncture, moxibustion, acupoint catgut embedding, acupoint application, and nasal irrigation with TCM, which are safe, simple, acceptable by patients and have good therapeutic effect. Finally, the clinical research status of TCM treatment of AR was summarized, and suggestions were put forward from three aspects: standardizing the clinical research protocol of TCM, encouraging pure TCM research, and exploring the mechanism of TCM treatment on the basis of frontier research achievements. This review aims to provide higher-level evidence for subsequent clinical research and promote the research on dominant AR diseases. 
		                        		
		                        		
		                        		
		                        	
8.A Case Report of Blau Syndrome
Guozhuang LI ; Kexin XU ; Sen ZHAO ; Jianguo ZHANG ; Guixing QIU ; Ruifang SUI ; Tao WANG ; Min SHEN ; Xuejun ZENG ; Wei WANG ; Mingsheng MA ; Min WEI ; Xiao LONG ; Ke LYU ; Li HUO ; Lei XUAN ; Nan WU
JOURNAL OF RARE DISEASES 2023;2(4):547-553
		                        		
		                        			
		                        			Blau syndrome is a rare genetic disorder characterized by the a mix of granulomatous arthritis, uveitis, and dermatitis. Patients typically manifest multisystem involvement, including ocular, skin, and skeletal abnormalities. Blau syndrome is extremely rare, with a global incidence of less than one in a million among children. In this multidisciplinary consultation, we present a case of a 21-year-old young female patient having multisystemic involvement since early childhood. She was presented with multiple joint swelling, skin lesions, increased eye discharge, and accompanied by hypertension and arterial abnormalities, and received a diagnosis of uveitis. The patient had been receiving steroid treatment since the age of 6 and has tried various medications, with some improvement in joint swelling and ocular symptoms. Through this rare disease multidisciplinary consultation, we aim to provide guidance in the molecular diagnosis of the patient, multisystem assessment, and the selection and formulation of treatment plans. Additionally, we hope that by reporting this case, clinical physicians can gain a better understanding of the diagnosis and comprehensive treatment strategies for Blau syndrome, thereby improving the management and treatment of rare diseases.
		                        		
		                        		
		                        		
		                        	
9.Effects of Acupuncture on Gut Microbiota and Central Iinflammation in Rat Model of Migraine based on"Gut-Brain"Theory
Mingsheng SUN ; Xingyu CHEN ; Mengdi ZHOU ; Yi LIU ; Yuqi ZHENG ; Siying HUANG ; Dingjun CAI ; Ling ZHAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(8):2726-2734
		                        		
		                        			
		                        			Objective Based on the theory of"gut-brain",this study explored the effect of acupuncture on the gut microbiota and central inflammation in migraine model rats,in order to explore the mechanism of acupuncture in the treatment of migraine from the perspective of"gut-brain".Methods The migraine rat model was established by subcutaneous injection of nitroglycerin.They were randomly divided into a model group and an acupuncture group,with 6 rats in each group,and a control group with 6 rats for conventional binding and fixation.Before modeling and on the 1st,5th,and 9th days after modeling,each group used electronic VonFrey to measure the plantar mechanical pain threshold of rats.After the experiment,Elisa was used to detect the expression levels of inflammatory factors IL-6 and TNF-α in the central trigeminal spinal nucleus of the rats in each group.Three-generation Pacbio full-length microbial diversity sequencing was used to perform 16S full-length rDNA sequencing on each group of fecal samples to detect the operational taxonomic unit(OTU)clustering and its abundance,Alpha diversity index,Beta diversity index,species among the samples in each group.differences in abundance.Results In migraine model rats,plantar mechanical pain threshold was significantly decreased(P<0.01),central IL-6 and TNF-α contents were significantly increased(P<0.01),and the structure and abundance of gut microbiota were abnormal.change(P<0.01).Continuous acupuncture treatment can significantly increase the plantar mechanical pain threshold in migraine rats(P<0.01),regulate the diversity of gut microbiota in migraine rats,increase Lactobacillus murine,and reduce the abundance of Lactobacillus enterobacteriaceae.degree(P<0.05),and decreased the levels of IL-6 and TNF-α in the central nervous system of migraine model rats(P<0.01).Conclusion Acupuncture can exert the"gut-brain"anti-inflammatory and analgesic effect of acupuncture in the treatment of migraine by regulating the gut microbiota structure and the expression of central IL-6 and TNF-α inflammatory factors in migraine model rats.
		                        		
		                        		
		                        		
		                        	
10.An Adolescent with Recurrent Intracranial Hemorrhage, and Skin Lesion
Yuehui HONG ; Min SHEN ; Tao WANG ; Mingsheng MA ; Sen ZHAO ; Feng FENG ; Dachun ZHAO ; Wen ZHANG ; Xuejun ZENG ; Lei XUAN ; Ming YAO ; Yicheng ZHU
JOURNAL OF RARE DISEASES 2022;1(2):151-157
		                        		
		                        			
		                        			We presented an adolescent with recurrent intracranial hemorrhage and skin lesion. The diagnosis was unclear and the treatment was difficult. Through a multidisciplinary effort type Ⅰ interferon disease was suspected and later, an interferon-stimulated gene was further detected. Considering the high morbidity and fatality rate of recurrent intracranial hemorrhage, tofacitinib and hydroxychloroquine were administered. After treatment, the livedo reticularis was significantly regressed. Unfortunately, the intracranial hemorrhage recurred due to a pre-existing cerebral aneurysm, leading to death of the patient. The diagnosis and treatment of this case highlight the importance of multidisciplinary collaboration in the diagnosis and treatment of difficult and rare diseases.
		                        		
		                        		
		                        		
		                        	
            
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