1.Exploring Mechanism of Different Processed Products of Baiyaojian and Its Formula in Treating Ulcerative Colitis Based on Serum Metabolomics and Flow Cytometry
Ruisheng WANG ; Yafang HOU ; Wenwen CAO ; Yahong ZHAO ; Zhenling ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):172-181
ObjectiveTo explore the potential mechanism of different processed products of Baiyaojian and its compound Xiangmei pills in rats with ulcerative colitis(UC) by comparing the pharmacodynamic and metabolomic differences. MethodEighty SD rats were acclimatized and kept for 3 d, and randomly divided into 8 groups[blank group, model group, mesalazine group(0.4 g·kg-1), Baiyaojian group(1.89 g·kg-1), stir-fried Baiyaojian group(1.89 g·kg-1), carbonized Baiyaojian group(1.89 g·kg-1), and Xiangmei pills low and high dose groups(1.89, 5.67 g·kg-1)], with 10 rats in each group. Rats in the blank group were administered physiological saline by gavage, and rats in the remaining 7 groups were orally administered 5% dextran sodium sulfate(DSS) solution daily for 8 consecutive days to induce UC model. After successful modeling, each dosing group was given the corresponding dose of drug solution by gavage, and the blank and model groups were given equal amounts of saline by gavage, and the drug was administered continuously for 8 d. Then serum levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-6, IL-10 and IL-1β were measured by enzyme-linked immunosorbent assay(ELISA), hematoxylin-eosin(HE) staining was used to observe the histopathological changes of colon tissue, the proportion of T helper 17 cells(Th17) and regulatory T cells(Treg) in the peripheral blood of rats in each group was detected by flow cytometry. The endogenous metabolites in serum of rats were detected by ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS), and the differential metabolites were characterized by combining principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA), and were analyzed according to the variable importance in the projection(VIP) value>1.0 and P<0.05, and potential metabolic pathways were analyzed according to Human Metabolome Database(HMDB). ResultCompared with the blank group, the colon tissue of the model group was congested and the mucosa was ulcerated, the colon length was significantly reduced(P<0.01) and the quality was significantly increased(P<0.05), the proportion of Th17/Treg in the peripheral blood and the serum levels of TNF-α, IL-6 and IL-1β were significantly increased, while the IL-10 expression wassignificantly reduced(P<0.05, P<0.01). Compared with the model group, the colon tissue of UC rats in each treatment group was improved with scattered ulcers, reduced inflammatory cell infiltration, significantly increased colon length, and significantly decreased mass(P<0.05), the proportion of Th17/Treg in the peripheral blood decreased, the expression of TNF-α,IL-6 and IL-1β was significantly reduced(P<0.05, P<0.01), while the IL-10 expression was significantly increased(P<0.01). The therapeutic effect of different administration groups on UC was in the order of high dose group of Xiangmei pills>low dose group of Xiangmei pills>carbonized Baiyaojian group>stir-fried Baiyaojian group>Baiyaojian group. And a total of 26 differential metabolites were screened in the metabolomics results. Compared with the blank group, 14 differential metabolites were up-regulated and 5 metabolites were down-regulated in the model group, and 14, 9, 14, 12 and 17 metabolites could be recalled in the Baiyaojian group, stir-fried Baiyaojian group, carbonized Baiyaojian group, Xiangmei pills low and high dose groups. The main metabolic pathways involved were citrate cycle pathway, pantothenic acid and coenzyme A biosynthesis pathway, aromatic hydrocarbon receptor(AhR) signaling pathway, glycolysis/gluconeogenesis pathway. ConclusionThe therapeutic effect of Baiyaojian on UC is significantly improved after charcoal stir-frying, and the efficacy is more prominent when combined with Angelicae Dahuricae Radix and Mume Fructus Carbonisata, which can provide a basis for the development of Baiyaojian compound preparations.
2.Identification and optimization of peptide inhibitors to block VISTA/PSGL-1 interaction for cancer immunotherapy.
Xiaoshuang NIU ; Menghan WU ; Guodong LI ; Xiuman ZHOU ; Wenpeng CAO ; Wenjie ZHAI ; Aijun WU ; Xiaowen ZHOU ; Shengzhe JIN ; Guanyu CHEN ; Yanying LI ; Jiangfeng DU ; Yahong WU ; Lu QIU ; Wenshan ZHAO ; Yanfeng GAO
Acta Pharmaceutica Sinica B 2023;13(11):4511-4522
Developing new therapeutic agents for cancer immunotherapy is highly demanding due to the low response ratio of PD-1/PD-L1 blockade in cancer patients. Here, we discovered that the novel immune checkpoint VISTA is highly expressed on a variety of tumor-infiltrating immune cells, especially myeloid derived suppressor cells (MDSCs) and CD8+ T cells. Then, peptide C1 with binding affinity to VISTA was developed by phage displayed bio-panning technique, and its mutant peptide VS3 was obtained by molecular docking based mutation. Peptide VS3 could bind VISTA with high affinity and block its interaction with ligand PSGL-1 under acidic condition, and elicit anti-tumor activity in vivo. The peptide DVS3-Pal was further designed by d-amino acid substitution and fatty acid modification, which exhibited strong proteolytic stability and significant anti-tumor activity through enhancing CD8+ T cell function and decreasing MDSCs infiltration. This is the first study to develop peptides to block VISTA/PSGL-1 interaction, which could act as promising candidates for cancer immunotherapy.
3.Gabapentin alleviates myocardial ischemia-reperfusion injury through the PI3K-AKT signaling pathway
Bin WU ; Yahong CAO ; Rui LI
Chinese Journal of Emergency Medicine 2022;31(3):344-348
Objective:To evaluate the effect of gabapentin on myocardial ischemia-reperfusion injury and its mechanism.Methods:Sixty male clean SD rats, aged 10 weeks and weighing 250 g~300 g, were divided into 5 groups ( n=12) with 12 rats in each group by random number table method: Sham group, myocardial ischemia reperfusion group (group I/R), gabapentin group (group Gap), LY294002 group (group LY) and gabapentin +LY294002 group (group Gap +LY). The model of myocardial ischemia reperfusion injury was established by ligation of the left anterior descending coronary artery for 30 min and reperfusion for 120 min. Heart rate (HR), mean arterial pressure (MAP) and the rate pressure product (RPP) were recorded at baseline before ischemia (T 0) for 30 min (T 1) and reperfusion for 120 min (T 2) to evaluate hemodynamic changes during ischemia and reperfusion; The frequency of PVCs and VT/VF were recorded to evaluate the occurrence of arrhythmias during ischemia/reperfusion. TTC staining was used to detect myocardial infarction area. And the protein expression levels of PI3K and Akt in myocardial tissue were detected by Western blotting. Results:Compared with group I/R, the myocardial infarction area, PVCs and VT/VF times, and the protein expression levels of PI3K and p-Akt were significantly increased in group Gap ( P<0.05). Compared with group Gap, the area of myocardial infarction, the number of PVCs and VT/VF, and the protein expression of PI3K and p-Akt were significantly decreased in the group Gap +LY ( P<0.05). Conclusions:Gabapentin can alleviate myocardial ischemia-reperfusion injury, and its mechanism is related to the activation of PI3K-AKT signaling pathway.
4.Application of lncRNA AK057037 in tissues and blood samples of patient with hepatocellular cancer
Yinchao FAN ; Yanan CAO ; Yahong WU ; Xiaosong BAI
International Journal of Laboratory Medicine 2018;39(6):664-667
Objective To investigate the clinical value of tissue and blood long noncoding RNA(lncRNA) detection in the patients with hepatocellular cancer(HCC).Methods The HCC tissue sample,blood sample, suspected HCC puncture sample and blood sample from the persons undergoing healthy physical examination were selected for conducting the study.The six lncRNAs which were more than 5-fold expression amount were picked out in the 338 transcripts with abnormal high expression.The expression amount of lncRNA AK057037 in HCC tissue and serm tissue was detected by using the real time quantitative polymerase chain reaction and in situ hybridization.Then the results were statistically analyzed.Results The six lncRNAs were highly ex-pressed in the tissues and blood of the liver cancer,and the difference was statistically significant(P<0.05). Compared with healthy people,the relative expression of AK057037 in the tissues and blood of the liver cancer increased significantly,and the relative expression level in tumor and paracancerous tissues was statistically significant(P<0.05).As the tumor progresses,the relative expression of AK057037 was further enhanced. There was a higher relative expression in advanced liver cancer especially,and the difference was statistically significant(P<0.05).The results of the receiver operating characteristic curve showed that the area under curve of AK057037 was 0.834(95% CI 0.705-0.918)in the blood specimen,and its diagnostic efficiency was better than that of commonly used nucleic acid marker miR-29a.Conclusion lncRNA AK057037 is highly ex-pressed in HCC sample and blood,and can be used for clinical detection of HCC.
5.Current status of allergy diagnostic testing in vitro of type Ⅰ allergy
Lei SHI ; Yahong CAO ; Jue ZHANG
Chinese Journal of Laboratory Medicine 2018;41(11):889-892
Type Ⅰallergy is one type of allergic reaction mediated by IgE antibody .Simple, rapid and accurate allergen detection method can be used to identify the allergen of patients , which is of great significance for the prevention , diagnosis and treatment of allergic diseases . In this article , different techniques for in vitro diagnosis of allergen specific IgE antibodies in type Ⅰ allergic reactions were described , important factors affecting in vitro diagnosis of allergens were analyzed , and the future development trend of in vitro diagnosis of allergen specific IgE antibodies were prospected .In order to help clinicians to select more suitable in vitro detection methods in the process of in vitro diagnosis of type Ⅰallergy and to better understand the internal relationship between the allergen specific IgE detection report and clinical symptoms.
6.Therapeutic effect of nasogastric tube and nasointestinal tube enteral nutrition for severe traumatic brain injury
Jing NIE ; Haihua CAO ; Yahong HOU ; Sai ZHANG
Chinese Journal of Trauma 2017;33(6):551-554
Objective To investigate the therapeutic effect of enteral nutrition in patients with severe traumatic brain injury (sTBI) by nasogastric tube and nasointestinal tube.Methods A retrospective case control study was made on 64 patients with sTBI hospitalized from October 2013 to December 2015.The patients were assigned to nasogastric tube group (n=32) and nasointestinal tube group (n=32) according to the random number table, and the same nutrient solution was given to all patients.Eighteen male and 14 female patients aged (37.8±8.7)years were identified in nasogastric tube group, and the Glasgow coma score (GCS) was (4.7±1.3)points.Twenty males and 14 females aged (39.7±6.5)years were identified in nasointestinal tube group, and the GCS was (4.1±1.2)points.Triceps skinfold thickness, body mass index, hemoglobin, level of serum albumin and level of prealbumin were measured and compared between groups before and after operation.Incidence of complications was recorded as well.Results All the above-mentioned indexes of the two groups did not differ significantly before the therapy (P>0.05), but showed significant differences 7 and 15 days after therapy (P<0.05).Incidence of complications in nasointestinal tube group (17 lung infection, nine gastrointestinal bleeding, 11 electrolyte imbalance) was lower than that in nasogastric tube group (seven lung infection, two gastrointestinal bleeding, one reflux esophagitis, four electrolyte imbalance) (P<0.05).Conclusion Nasointestinal tube enteral nutrition is better than nasogastric tube enteral nutrition in improving nutritional status and reducing complications in patients with sTBI.
7.G1986A and G1899A in the pre-C region of HBV promote the serological conversion of e antigen
Huawen YANG ; Yan ZHU ; Yachao YAO ; Yahong LI ; Nan LI ; Donglin CAO ; Zhi ZHANG ; Liangshan HU
The Journal of Practical Medicine 2017;33(6):990-993
Objective To investigate the correlation of the 1896 and 1899 mutations of hepatitis B virus (HBV)with the conversion of e antigen in serum and the progression of the disease. Methods 238 serum samples from the patients with HBsAg positive for over six months and HBV-DNA copy number > 5.0 × 102 IU/mL were collected,and the sequence analysis was used to analyze the nucleotide sequences of the 1896 and 1899 sites in the pre-C region of HBV. At the same time,the relevant clinical data and the expressions of HBeAg were collected,followed by Spearman correlation analysis and chi square test with SPSS 20.0. Results Both 1896 and 1899 sites in the pre-C region of HBV were mutated,and the base G was A,which was closely related to the expression of e antigen(P<0.05). Both G1896A and G1899A promoted the e antigen serological conversion ,and the e antigen serological conversion of G1899A was higher than that of G1896A. G1899A was associated with HBV related disease progression (correlation coefficient 0.280,P < 0.05),especially with the incurrence of HCC. Conclusions G1896A and G1899A in the pre-C region of HBV can promote the serological conversion of e antigen.
8.Efficacy analysis of acupuncture with biofeedback in the treatment of patients with functional anorectal pain.
Ling ZHENG ; Shuqing DING ; Yijiang DING ; Yahong XUE ; Huifen ZHOU ; Min LI ; Jianbao CAO ; Jing WANG
Chinese Journal of Gastrointestinal Surgery 2016;19(12):1375-1378
OBJECTIVETo observe the short- and long-term efficacy of acupuncture combined with biofeedback in the treatment of functional anorectal pain (FARP).
METHODSClinical data of 142 patients who met the functional gastrointestinal disorders and functional anorectal pain based on criteria of Rome III( undergoing acupuncture with biofeedback therapy from August 2010 to November 2015 in Pelvic Floor Center of The Third Affiliated Hospital of Nanjing University of Chinese Medicine were retrospectively analyzed. Telephone and outpatient clinic recheck were used as standard follow-up. The clinical effect of short-term and long-term data collected from the disease-based database was evaluated with visual analogue pain scale (VAS) (0-10 points), short form health survey questionnaire (SF-36) (0-148 points). The overall satisfaction and effectiveness (VAS was >30%) were evaluated at the end of treatment (short-term) and during follow-up (long-term).
RESULTSThe effective follow-up data were obtained from 71.1%(101/142) of patients and the median follow-up time was 28(3-67) months. The VAS of 101 cases was 6.09±1.78, 1.99±1.89 and 3.55±2.60 before treatment, at the end of treatment and during follow-up respectively. Though the VAS during follow-up was higher than that at the end of treatment, but still significantly lower than that before treatment(P<0.05). The SF-36 score of 31 patients was 82.0±16.9, 94.0±15.1 and 88.1±15.3 before treatment, at the end of treatment and during follow-up respectively. Though the SF-36 score during follow-up was lower compared to at the end of treatment, but still significantly higher compared to before treatment (P<0.05). The effective rates were 85.9%(122/142) at the end of treatment and 75.2%(76/101) during follow-up, and the satisfactory rates were 92.3%(131/142) and 84.2%(85/101), respectively.
CONCLUSIONAcupuncture with biofeedback has significant short-term and long-term efficacy in treating functional anorectal pain, and its degree of satisfaction is high.
9.Effect of Acupuncture on HDAC2 Activity in Peripheral Blood of COPD Rats
Jie ZHANG ; Yahong CAO ; Xiaobo ZHU ; Mingming ZHOU ; Xinhui ZHU ; Wei LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;17(4):861-864
This article was aimed to explore the effect of acupuncture on HDAC2 activity in peripheral blood of chronic obstructive pulmonary disease (COPD) rats. The COPD rat model was established by cigarette smoking. Acupuncture was given for 3 consecutive weeks. The lung ventilation of rat was detected by equipment. Peripheral blood of rat was taken after 3 weeks. The HDAC2 enzyme activity was detected by fluorescence technology. ELISA was used in the detection of IL-10, IL-8 and TNF-αexpression in peripheral blood. The results showed that after acupuncture treatment, the lung ventilation was obviously increased compared with the model group (P < 0.05). Meanwhile, the HDAC2 activity in the peripheral blood of rat was obviously increased (P < 0.05). However, the expression of inflammatory cytokines IL-10, IL-8 and TNF-α was obviously reduced (P < 0.05). It was concluded that acupuncture treatment had a good efficacy on COPD rat. It indicated that acupuncture treatment might regulate HDAC2 activity to reduce inflammation.
10.The diagnostic value of pelvic floor sEMG in pelvic floor dyssynergiaby using receiver operating characteristic curve (ROC curve)
Yahong XUE ; Shuqing DING ; Yijiang DING ; Jing WANG ; Min LI ; Jianbao CAO ; Min NI
The Journal of Practical Medicine 2014;(22):3586-3588
Objective To evaluate the clinical diagnostic value of pelvic floor sEMG in pelvic floor dyssynergia (PFD) by using receiver operating characteristic curve (ROC curve). Methods The pelvic floor sEMG of 90 patients with PFD and 101normal controls were determined by the Glazer protocol.Parameters including amplitude (AVG),coefficient of variance (CV),onset time and median frequency (MF),and the ROC curve werealso investigated. Results Compared to the control group,the PFD group had a higher AVG of pre-baseline (P < 0.05), a lower AVG during Flick and Tonic steps(P < 0.05), and ahigher CV duringTonic and Endurance steps(P < 0.05).The area under curve(AUC) of CV duringthe tonic step was 0.883 withthe best cut-off of 0.355, and with sensitivity of 88.4%and the specificity of 71.1%, respectively; The AUC of CV duringEndurance step was 0.825 withthe best cut-off of 0.305, and with the sensitivity of 84.9%and the specificity of 67.8% , respectively. Conclusion The CVs of the tonic and the endrnace phases can be used as valuable clinical values in diagnosis of PFD.

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