1.Mechanism of Naoxintong Capsules Against Ischemia-reperfusion Injury in Rats via Inhibiting Pericyte Contraction Based on RHOA/ROCK1 Pathway
Yinlian WEN ; Jinfeng SHANG ; Bohong WANG ; Wanting WEI ; Xiaolu ZHANG ; Guijinfeng HUANG ; Xin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):159-167
ObjectiveTo investigate the mechanism of Naoxintong capsules on ischemia-reperfusion (I/R) injury in rats based on the changes of pericytes mediated by Ras homolog family member A (RHOA)/Rho-associated coiled-coil containing protein kinase 1 (ROCK1) pathway. MethodsNinety rats (15 rats for each group) were randomly divided into a sham operation group, a model group, a positive control group receiving Ginkgo biloba extract (21.6 mg·kg-1), and groups receiving Naoxintong capsules at low, medium, and high doses of 55, 110, and 220 mg·kg-1 (NXT-L, NXT-M, and NXT-H groups), respectively. Except for those in the sham operation group, all rats were subjected to transient middle cerebral artery occlusion (tMCAO) to establish the experiment model. Nerve function was assessed using a neurological function score. Cerebral blood flow was detected using a laser speckle contrast imager, and the cerebral infarction rate was calculated using 2,3,5-Triphenyl tetrazolium chloride (TTC) staining. Pathological changes were observed by hematoxylin-eosin (HE) staining and Nissl staining, while pericyte morphology was observed via transmission electron microscopy. Blood-brain barrier destruction was observed by Evans blue staining. Albumin and ischemia-modified albumin levels were measured using assay kits. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to detect the mRNA and protein expression levels of RHOA, ROCK1, platelet-derived growth factor receptor β (PDGFRB), α-smooth muscle actin (α-SMA), tight junction protein (ZO-1), matrix metalloproteinase-2 (MMP-2), and matrix metalloproteinase-9 (MMP-9). ResultsCompared with the sham operation group, the model group exhibited decreased neurological function scores, higher percentage reduction in blood flow, and increased cerebral infarction rates (P<0.01). Additionally, cortical neuronal nucleus shrinkage, edema, a decreased number of Nissl bodies, reduced pericyte area, elevated albumin content in the cortex (P<0.05), and increased ischemic modified albumin levels (P<0.01) were observed. The mRNA and protein expression levels of RHOA, ROCK1, PDGFRB, α-SMA, MMP-2, and MMP-9 were increased (P<0.01), while those of ZO-1 were decreased. Compared with the model group, all treatment groups showed improved neurological function scores, lower percentage reduction in blood flow, reduced cerebral infarction rates (P<0.01), alleviated cortical histological changes, increased number of Nissl bodies, expanded pericyte area, decreased albumin content in the cortex, and reduced ischemia-modified albumin levels (P<0.01). The mRNA and protein expression levels of RHOA, ROCK1, PDGFRB, α-SMA, MMP-2, and MMP-9 were decreased (P<0.01), while those of ZO-1 were increased. Among the treatment groups, the NXT-M group showed the most pronounced improvement in cerebral I/R injury. ConclusionNaoxintong capsules can restore cerebral blood supply, reduce microcirculation disturbance, and protect blood-brain barrier in rats with I/R injury. Its mechanism of action may be related to the inhibition of the RHOA/ROCK1 signaling pathway and reduced pericyte contraction.
2.Mechanism of Naoxintong Capsules Against Ischemia-reperfusion Injury in Rats via Inhibiting Pericyte Contraction Based on RHOA/ROCK1 Pathway
Yinlian WEN ; Jinfeng SHANG ; Bohong WANG ; Wanting WEI ; Xiaolu ZHANG ; Guijinfeng HUANG ; Xin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):159-167
ObjectiveTo investigate the mechanism of Naoxintong capsules on ischemia-reperfusion (I/R) injury in rats based on the changes of pericytes mediated by Ras homolog family member A (RHOA)/Rho-associated coiled-coil containing protein kinase 1 (ROCK1) pathway. MethodsNinety rats (15 rats for each group) were randomly divided into a sham operation group, a model group, a positive control group receiving Ginkgo biloba extract (21.6 mg·kg-1), and groups receiving Naoxintong capsules at low, medium, and high doses of 55, 110, and 220 mg·kg-1 (NXT-L, NXT-M, and NXT-H groups), respectively. Except for those in the sham operation group, all rats were subjected to transient middle cerebral artery occlusion (tMCAO) to establish the experiment model. Nerve function was assessed using a neurological function score. Cerebral blood flow was detected using a laser speckle contrast imager, and the cerebral infarction rate was calculated using 2,3,5-Triphenyl tetrazolium chloride (TTC) staining. Pathological changes were observed by hematoxylin-eosin (HE) staining and Nissl staining, while pericyte morphology was observed via transmission electron microscopy. Blood-brain barrier destruction was observed by Evans blue staining. Albumin and ischemia-modified albumin levels were measured using assay kits. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to detect the mRNA and protein expression levels of RHOA, ROCK1, platelet-derived growth factor receptor β (PDGFRB), α-smooth muscle actin (α-SMA), tight junction protein (ZO-1), matrix metalloproteinase-2 (MMP-2), and matrix metalloproteinase-9 (MMP-9). ResultsCompared with the sham operation group, the model group exhibited decreased neurological function scores, higher percentage reduction in blood flow, and increased cerebral infarction rates (P<0.01). Additionally, cortical neuronal nucleus shrinkage, edema, a decreased number of Nissl bodies, reduced pericyte area, elevated albumin content in the cortex (P<0.05), and increased ischemic modified albumin levels (P<0.01) were observed. The mRNA and protein expression levels of RHOA, ROCK1, PDGFRB, α-SMA, MMP-2, and MMP-9 were increased (P<0.01), while those of ZO-1 were decreased. Compared with the model group, all treatment groups showed improved neurological function scores, lower percentage reduction in blood flow, reduced cerebral infarction rates (P<0.01), alleviated cortical histological changes, increased number of Nissl bodies, expanded pericyte area, decreased albumin content in the cortex, and reduced ischemia-modified albumin levels (P<0.01). The mRNA and protein expression levels of RHOA, ROCK1, PDGFRB, α-SMA, MMP-2, and MMP-9 were decreased (P<0.01), while those of ZO-1 were increased. Among the treatment groups, the NXT-M group showed the most pronounced improvement in cerebral I/R injury. ConclusionNaoxintong capsules can restore cerebral blood supply, reduce microcirculation disturbance, and protect blood-brain barrier in rats with I/R injury. Its mechanism of action may be related to the inhibition of the RHOA/ROCK1 signaling pathway and reduced pericyte contraction.
3.Screening of anti-COPD quality biomarkers of Gnaphalium affine
Wanting LIU ; Rong XIE ; Dahuai LIN ; Xiangli YE ; Guohong YAN ; Huang LI
China Pharmacy 2024;35(6):683-688
OBJECTIVE To screen the quality biomarkers of Gnaphalium affine with anti-chronic obstructive pulmonary disease (COPD) effect and determine their contents. METHODS The effective components and targets of “G. affine” with anti- COPD effect were predicted by using network pharmacology as a search criterion. HPLC fingerprints for 10 batches of G. affine were established by using Similarity Evaluation System of TCM Chromatographic Fingerprint (2012 edition); common peak identification and similarity evaluation were conducted; cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) were performed to screen differential components as quality maker that affected the quality of G. affine using variable importance projection (VIP)>1 as the standard. The same HPLC method was adopted to determine the contents of the differential components in 10 batches of samples. RESULTS A total of 10 flavonoids (such as quercetin, luteolin, and chlorogenic acid) and organic acid components, were identified through network pharmacology search, with 91 targets closely related to anti-COPD. A total of 9 common peaks were identified in 10 batches of samples, with similarity greater than 0.90. Among them, the differential components included chlorogenic acid, caffeic acid, 1,3-O- dicaffeoylquinic acid and apigenin 7-O-β-D-glucopyranoside; S3, S4, S6, S7 and S10 were clustered into one category, S2, S5, S8 and S9 clustered into one category, and S1 clustered into one category. The contents of chlorogenic acid, caffeic acid, 1,3-O- dicaffeoylquinic acid, and apigenin 7-O-β-D-glucopyranoside in 10 batches of G. affine ranged 0.070-7.653, 0.010-0.097, 0.001- 0.036, 0.508-6.627 mg/g, respectively. CONCLUSIONS Chlorogenic acid, caffeic acid, 1,3-O-dicaffeoylquinic acid, apigenin 7- O-β-D-glucopyranoside can serve as the potential quality marker for the anti-COPD effect of G. affine, with the highest content of chlorogenic acid in G. affine produced in Ji’an, Jiangxi province, and the highest content of caffeic acid in G. affine produced in Ji’an, Jiangxi province and Sanming, Fujian province. The contents of the last two components are highest in G. affine produced in Chaoshan, Guangdong province.
4.Clinical Efficacy of TCM Multi-channel Intervention Plus Antagonist Regimen on Ovarian Function,TCM Syndromes and Natural Conception in Poor Ovarian Responders Undergoing IVF-ET
Yunji XU ; Wanting XIA ; Xuan ZHANG ; Qianhong MA ; Jinzhu HUANG ; Qian ZENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):93-100
ObjectiveTo objectively analyze the effects of traditional Chinese Medicine (TCM) multi-channel intervention on the ovarian function,TCM syndromes and natural conception of poor ovarian responders(kidney-Yin deficiency,liver depression and blood stasis pattern) who planned to receive another in vitro fertilization embryo transfer(IVF-ET)antagonist regimen. MethodThe 128 low-prognosis patients (kidney Yin deficiency,liver depression and blood stasis pattern) who attended the West China Second University Hospital, Sichuan University and the Hospital of Chengdu University of Traditional Chinese Medicine from August 2020 to February 2023 and met the inclusion criteria were selected,and then divided into the treatment group and the control group according to the random number table,with 64 patients in each group. The control group was treated with oral dehydroepiandrosterone(DHEA),while the treatment group was treated with multi-channel TCM(oral TCM decoction + auricular point sticking + Bushen Huoxue prescription through retention enema). After 3 menstrual cycles,the relevant indicators for ovarian function evaluation,TCM syndrome scores and natural conception were collected from both groups. ResultCompared with the situation before treatment,the basal follicle stimulating hormone(bFSH),bFSH/basal luteinizing hormone(bLH),basal estradiol(bE2),antral follicle count(AFC),the number of oocytes obtained,the number of normal fertilization,the number of superior embryos and TCM syndrome scores in the treatment group were improved after treatment(P<0.05,P<0.01). For the control group, the bFSH/bLH and TCM syndrome scores were increased after treatment(P<0.05), while the bFSH,bFSH/bLH,bE2,AFC,the number of oocytes obtained,the number of normal fertilization,and the number of superior embryos showed no significant difference after treatment. Compared with the control group after treatment,bFSH,bFSH/bLH,bE2,AFC,the number of normal fertilization,the number of superior embryos and TCM syndrome scores in the treatment group were better (P<0.05,P<0.01),while there was no significant difference in the number of oocytes obtained. After treatment,there were 3 cases of natural conception in the treatment group,while there were no natural conception in the control group. ConclusionFor patients with poor ovarian response and kidney Yin deficiency,liver depression and blood stasis pattern,multi-channel intervention of TCM plus the antagonist regimen can reduce bFSH,bFSH/bLH values,improve the levels of bE2,increase AFC,the number of oocytes obtained,the number of normal fertilization and the number of superior embryos,improve ovarian function,menstruation and TCM syndromes,improve their quality of life,and even enable some patients to get pregnant naturally before re-progression and improve their pregnancy outcome.
5.Combined Therapies of Traditional Chinese Medicine Treat Low-prognosis Patients Undergoing IVF-ET Again
Xuan ZHANG ; Wanting XIA ; Jinzhu HUANG ; Qianhong MA ; Yunji XU ; Qian ZENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(1):160-169
ObjectiveTo objectively evaluate the clinical efficacy of multiple therapies of traditional Chinese medicine (TCM) in low-prognosis patients who received antagonist protocol for in vitro fertilization and embryo transfer (IVF-ET) again. MethodA total of 128 patients with kidney Yin deficiency, liver depression, and blood stasis who planned to receive antagonist protocol for IVF-ET in the West China Second Hospital of Sichuan University were enrolled and assigned into two groups by random number table method. The observation group (64 casces) was treated by oral administration of Chinese medicine decoction + enema of kidney-tonifying and blood-activating method + auricular point sticking + oral administration of dehydroepiandrosterone (DHEA), while the control group (64 casces) was treated by only oral administration of DHEA. After treatment for three menstrual cycles, both groups received the antagonist protocol for IVF-ET. The TCM syndrome scores, basic sex hormone levels, antral follicle count (AFC), the usage of gonadotropin (Gn), endometrial receptivity indicators, embryo quality indicators, and pregnancy outcomes were compared between the two groups. ResultAfter treatment, the observation group showed decreased follicle-stimulating hormone (FSH)/luteinizing hormone (LH) ratio, lowered level of estradiol (E2), increased AFC, decreased amount and days of Gn usage, improved endometrial receptivity indicators (endometrial thickness on trigger and ET days, proportion of endometrial type A in endometrial types and the level of E2 on trigger day) and embryo quality indicators (the rates of mature follicles, fertilization, normal fertilization, and premium embryos), and decreased TCM syndrome scores (P<0.05, P<0.01). Moreover, the observation group had lower FSH/LH ratio, E2 level, and amount of Gn usage, higher AFC, poorer endometrial receptivity and embryo quality indicators, and lower TCM syndrome scores than the control group after treatment (P<0.05, P<0.01). In addition, except for 3 cases of natural pregnancy, the observation group outperformed the control group in terms of improving the clinical pregnancy rates during initiation cycle and transplantation cycle and clinical pregnancy rate and decreasing biochemical pregnancy rate and early abortion rate (P<0.05). ConclusionCombined therapies of TCM can alleviate the clinical symptoms, reduce TCM syndrome scores, reduce the Gn usage amount, improve the number and quality of embryos and endometrial receptivity, and coordinate the synchronous development of endometrium and embryo. In this way, they can increase the clinical pregnancy rate and reduce biochemical pregnancy rate and early abortion rate in the low prognosis patients with kidney yin deficiency, liver depression, and blood stasis who are undergoing IVF-ET again.
6.Transcriptome Sequencing on Treatment of Kidney Deficiency and Blood Stasis-thin Endometrium in Rats with Bushen Huoxue Prescription Through Enema
Xuan ZHANG ; Wanting XIA ; Zhixing YIN ; Nana HAN ; Jinzhu HUANG ; Hang ZHOU ; Yi WANG ; Juan LI ; Qian ZENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(2):79-90
ObjectiveTo explore the mechanism of Bushen Huoxue enema in treating the rat model of kidney deficiency and blood stasis-thin endometrium (KDBS-TE) by transcriptome sequencing. MethodThe rat model of KDBS-TE was established by administration of tripterygium polyglycosides tablets combined with subcutaneous injection of adrenaline. The pathological changes of rat endometrium in each group were then observed. Three uterine tissue specimens from each of the blank group, model group, and Bushen Huoxue enema group were randomly selected for transcriptome sequencing. The differentially expressed circRNAs, lncRNAs, and miRNAs were screened, and the disease-related specific competitive endogenous RNA (ceRNA) regulatory network was constructed. Furthermore, the gene ontology (GO) functional annotation and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were performed for the mRNAs in the network. ResultCompared with the blank group, the model group showed endometrial dysplasia, decreased endometrial thickness and endometrial/total uterine wall thickness ratio (P<0.01), and differential expression of 18 circRNAs, 410 lncRNAs, and 7 miRNAs. Compared with the model group, the enema and estradiol valerate groups showed improved endometrial morphology and increased endometrial thickness and ratio of endometrial to total uterine wall thickness (P<0.05). In addition, 21 circRNAs, 518 lncRNAs, and 17 miRNAs were differentially expressed in the enema group. The disease-related specific circRNA-miRNA-mRNA regulatory network composed of 629 nodes and 664 edges contained 2 circRNAs, 34 miRNAs, and 593 mRNAs. The lncRNA-miRNA-mRNA regulatory network composed of 180 nodes and 212 edges contained 5 lncRNAs, 10 miRNAs, and 164 mRNAs. The mNRAs were mainly enriched in Hippo signaling pathway, autophagy-animal, axon guidance, etc. ConclusionBushen Huoxue enema can treat KDBS-TE in rats by regulating specific circRNAs, lncRNAs, and miRNAs in the uterus and the ceRNA network.
7.Discussion on the Idea of Moxibustion in the Treatment of Alzheimer's Disease Based on Thought of Paying Importance to Yang
Yuanyuan TONG ; Wanting ZHENG ; Shanrong HUANG ; Ling ZHU
International Journal of Traditional Chinese Medicine 2024;46(12):1537-1542
Alzheimer's disease (AD), a chronic degenerative disease of the central nervous system, is more prevalent in the elderly population and its incidence is increasing every year. It is important to pay attention to new methods of AD prevention and treatment for clinical treatment of AD, improving the quality of life of patients . This article analyzed the association between the theory and the etiology and pathogenesis of AD and the research basis of moxibustion for the prevention and treatment of AD based on the theory of "thought of emphasizing yang", in order to provide a broader idea for the TCM treatment of AD.
8.Clinical characteristics of 37 antiphospholipid syndrome patients complicated by autoimmune hemolytic anemia
Jintao ZHANG ; Wanting QI ; Yangzhong ZHOU ; Can HUANG ; Jiuliang ZHAO ; Mengtao LI ; Xiaofeng ZENG
Chinese Journal of Internal Medicine 2023;62(2):147-155
Objective:We sought to investigate the clinical characteristics and risk factors of antiphospholipid syndrome (APS) complicated by autoimmune hemolytic anemia (AIHA).Methods:Retrospective anaysis.Three hundred fifteen consecutive patients with APS were enrolled at the Department of Rheumatology of Peking Union Medical College Hospital between May 2017 to May 2021, and their clinical manifestations[including initial symptoms, time interval between APS onset and diagnosis, systemic lupus erythematosus(SLE), thrombotic events, obstetric morbidity, and extra-criteria manifestations] and laboratory test results[including blood routine, antiphospholipid antibodies(aPLs), blood lipid profile, homocysteine, anti-nuclear antibody profile, immunoglobulin levels, and complement levels] were collected. Then, univariate and multivariate logistic regression analyses were performed. Clinical features and risk factors were analyzed using univariable and multivariable logistic regression analysis.Results:Among 315 APS patients, 37 cases (11.7%) were complicated by AIHA, and AIHA was the first manifestation or co-occurrence. The median time interval between APS onset and diagnosis was 12 months. The proportion of SLE in APS patients combined with AIHA was higher than that in APS patients without AIHA[62.2%(23/37) vs. 19.4%(54/278), P<0.001]. There was no significant difference in the proportions of thrombosis and pregnancy morbidity between the two groups. In terms of extra-criteria manifestations, APS patients with AIHA had a significantly ( P<0.05) greater risk of thrombocytopenia ( OR=6.19, 95% CI 2.81-13.65) and higher proportions of hypocomplementemia, a positive lupus anticoagulant (LA) result, double aPLs positivity[i.e., any two of the following antibodies were positive: LA, anticardilolipin antibody(aCL), and anti-β2 glycoprotein Ⅰ(β2GPⅠ)], and triple aPLs positivity (i.e., LA, aCL, and anti-β2GPⅠ antibodies were all positive). Multivariate logistic regression analysis showed that SLE ( OR=3.46,95% CI 1.60-7.48), thrombocytopenia ( OR=2.56,95% CI 1.15-5.67), and hypocomplementemia ( OR=4.29,95% CI 2.03-9.04) were independent risk factors for the complication of APS. In the primary APS subgroup, multivariate logistic regression analysis showed that livedo reticularis ( OR=10.51,95%CI 1.06-103.78), thrombocytopenia ( OR=3.77, 95% CI 1.23-11.57), and hypocomplementemia ( OR=5.92,95% CI 1.95-17.95) were independent risk factors for the complication of APS. Conclusions:AIHA is not rare in APS patients; moreover, it occurs more frequently in APS secondary to SLE and is more likely to present with a variety of extra-criteria manifestations. Patients with AIHA should be promptly tested for antiphospholipid antibody profiles and alerted to the possibility of thrombotic events.
9.Research Advance in Primary Liver Cancer of qi Stagnation and Blood Stasis Pattern
Wanting HUANG ; Tao GUO ; Jing ZHANG ; Yong XIA ; Feng SHEN
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(11):3724-3727
Primary liver cancer(PLC)is one of the most common malignant tumors of digestive system in clinic.TCM plays an important role in the treatment of PLC.After sorting out the medical records and literatures related to the syndrome types of primary liver cancer in recent years,it was found that the syndrome type of qi stagnation and blood stasis was one of the important syndrome types of primary liver cancer.This article discusses and supplements the syndrome type of qi stagnation and blood stasis from the aspects of etiology and pathogenesis,proportion of syndrome types,staging of liver cancer,objective indicators of liver cancer,etc.It provides a new understanding for the treatment of primary liver cancer based on syndrome differentiation,so as to improve the clinical cure rate of liver cancer,alleviate the clinical symptoms of patients,improve the quality of life,and prolong the survival period.
10.Management of anticoagulant therapy in a patient with complex venous thromboembolism complicated with acute renal insufficiency
Yongjia HUANG ; Dequan JIANG ; Ling DING ; Jie LIU ; Xiaoyan HE ; Xiaoya LI ; Wanting HUANG ; Jindou LI
China Pharmacy 2023;34(18):2274-2277
OBJECTIVE To provide a reference for the safe use of drugs in patients with complex venous thromboembolism (VTE) and acute renal insufficiency. METHODS Clinical pharmacists participated in the management of anticoagulant therapy for a patient with complex VTE complicated with acute renal insufficiency, and evaluated the patient as high-risk thrombosis and bleeding based on their medical history, laboratory test results, etc.; combined with the complexity of thrombosis and renal insufficiency, clinical pharmacists suggested that enoxaparin sodium should be used in the acute stage of thrombosis (5 to 21 days after onset), and then warfarin should be adopted for oral anticoagulation treatment. Because the patient’s anticoagulation was not up to the standard (the target range of the international normalized ratio was 2-3), clinical pharmacists suggested increasing the warfarin dose, detecting the warfarin metabolism genotype, and adjusting the warfarin dose according to the genotype; at the same time, clinical pharmacists developed an anticoagulation monitoring plan to ensure the safety of anticoagulation treatment. RESULTS Doctors had adopted all the recommendations of clinical pharmacists. The patient did not experience adverse events such as bleeding or worsening of thromboembolism during anticoagulation in the hospital. When the anticoagulation met the standards, the patient was allowed to be discharged with medication. CONCLUSIONS By participating in the anticoagulation treatment management of patients with complex VTE and acute renal insufficiency, clinical pharmacists have assisted doctors in formulating personalized anticoagulation plans to promote the compliance with the anticoagulation treatment standard and ensure the safety and effectiveness of medication for patients.

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