1.Study on quality evaluation of Buddleja officinalis from different habitats based on HPLC fingerprint and content determination
Cuijie WEI ; Xiaoxia LIU ; Zhiwen DUAN ; Yongwei FENG ; Xiaozhou JIA ; Yueyi LIANG ; Xiangdong CHEN ; Dongmei SUN ; Zhenyu LI
International Journal of Traditional Chinese Medicine 2024;46(2):215-221
Objective:To establish UPLC fingerprint method and 2 contents determination methods of Buddleja officinalis; To provide a reference for improving the quality control standard and evaluation of Buddleja officinalis from different habitats.Methods:UPLC method was used to establish the fingerprints of 17 batches of Buddleja officinalis. The similarity evaluation, clustering analysis, principal component analysis and orthogonal partial least squares discriminant analysis were used to compare the quality differences of Buddleja officinalis from different habitats. The contents of acteoside and linarin in Buddleja officinalis were determined.Results:There were 12 common peaks in UPLC fingerprints of Buddleja officinalis, six of which were identified as echinacoside, acteoside, cynaroside, isoacteoside, linarin, and apigenin. The fingerprint similarity of 17 batches of Buddleja officinalis was more than 0.9; Buddleja officinalis from different habitats were classified into 2 groups. Five differential markers were determined by OPLS-DA analysis. The order of significance was acteoside > peak 3 > echinacoside > isoacteoside > linarin. Edgeworthia chrysantha was identified by the method of fingerprint as counterfeit. The results of content determination showed that the content of Buddleja officinalis in Hubei and Sichuan was the high and stable.Conclusion:The method can effectively analyze the differences of Buddleja officinalis from different habitats, and provide reference for the quality control of Buddleja officinalis.
2.Bibliometrics analysis of the application of diabetic retinopathy model
Zhuoyu HU ; Xiangdong CHEN ; Qi HU ; Zhimin LIU
International Eye Science 2024;24(12):1900-1907
AIM: To analyze the modeling characteristics and validation indexes of diabetic retinopathy model, analyze the shortcomings of the present animal experimental modeling, and provide a reference basis for the establishment of the standardization of the diabetic retinopathy model.METHODS: Literatures related to animal experiments on diabetic retinopathy were searched through the databases of CNKI, Wanfang, VIP and PubMed, and the experimental animal species, grade, gender, age, modeling method, modeling period, validation indexes, and other indexes were summarized and analyzed.RESULTS: The 275 papers that met the criteria were included. The animal models of diabetic retinopathy were mainly SD rats and Wistar rats, the sex of the experimental animals was mainly male, and the animal breeds were mostly of the SPF class. The age of most of the animals used was in the range of 6-8 weeks old; the modeling was based on those who established the type 1 diabetes model, mainly using STZ as the induction model. While the type 2 diabetes model was based on the high-fat, high-sugar diet combined with STZ. The modeling criteria were verified by detecting retinal morphology and structure, retinal vascularization, retinal function, and retinal cell apoptosis. In addition, the model was evaluated by detecting abnormal vascular proliferation, oxidative stress indicators, and inflammatory factor levels in retinal tissues, as well as abnormal vascular proliferation and inflammatory factor levels in aqueous humour, vitreous humor, and blood serum.CONCLUSION: Although the animal model of diabetic retinopathy has become a hotspot, the existing review is not comprehensive. Therefore, we summarized and analyzed the elements of the animal model through literature collation, including its characteristics and limitations, and providing methodological references for the establishment of the model, with a view to laying a solid foundation for the subsequent clinical and basic research of traditional Chinese medicine.
3.Effect of Modified Dahuang Huanglian Xiexintang on Mitochondrial Autophagy and Browning of Visceral Fat in Obese Type 2 Diabetes Mellitus Rats
Dong AN ; Yonglin LIANG ; Yankui GAO ; Fengzhe YAN ; Sichen ZHAO ; Zhongtang LIU ; Chengjun MA ; Xiangdong ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(24):131-140
ObjectiveTo observe the effect of modified Dahuang Huanglian Xiexintang on mitochondrial autophagy and browning of visceral adipose tissue in obese type 2 diabetes mellitus (T2DM) model ZDF rats. MethodForty ZDF rats were induced with a high-fat diet to establish an obese T2DM model. The rats were randomly divided into five groups: Model group, metformin group (0.18 g·kg-1), and high, medium, and low dose groups of modified Dahuang Huanglian Xiexintang (2.16, 1.08, 0.54 g·kg-1), with eight rats in each group. Additionally, eight ZDF (fa/+) rats were assigned to the normal group. All groups received an intragastric volume of 10 mL·kg-1, with the model and normal groups receiving the same volume of purified water once daily for 12 weeks. Fasting blood glucose (FBG) was regularly measured. After 12 weeks of intervention, the body weight, epididymal fat weight, and serum levels of glucose (GLU), glycated serum protein (GSP), triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were measured. Hematoxylin-eosin (HE) staining was used to observe pathological changes in epididymal fat tissue. Transmission electron microscopy (TEM) was employed to observe mitochondrial autophagy in adipocytes. Real-time PCR was used to detect the mRNA expression of hypoxia-inducible factor-1α (HIF-1α), Bcl-2/adenovirus E1B 19 kDa interacting protein 3 (BNIP3), microtubule-associated protein 1 light chain 3B (LC3B), p62/SQSTM1, uncoupling protein 1 (UCP1), iodothyronine deiodinase 2 (Dio2), and PR domain containing 16 (Prdm16) in epididymal fat. Western blot was used to detect the protein expression of HIF-1α, BNIP3, LC3B, p62, and UCP1 in epididymal fat. ResultCompared with the normal group, the model group showed pathological changes in epididymal fat, with adipocyte mitochondrial condensation and numerous autophagosomes indicating mitochondrial autophagy. The model group also exhibited significantly increased body weight, epididymal fat weight, FBG, GLU, GSP, TC, TG, and LDL-C levels (P<0.01), significantly decreased HDL-C levels (P<0.01), significantly elevated mRNA and protein expression of HIF-1α, BNIP3, and LC3B (P<0.01), significantly reduced mRNA and protein expression of p62 and UCP1 (P<0.01), and significantly reduced mRNA expression of Dio2 and Prdm16 (P<0.01). Compared with the model group, all intervention groups showed varying degrees of improvement in epididymal fat pathology. The metformin group and high-dose modified Dahuang Huanglian Xiexintang group displayed intact mitochondrial morphology, clear cristae, uniform matrix, and few autophagosomes and autophagosomes in the adipocyte cytoplasm. The metformin group and high- and medium-dose groups of modified Dahuang Huanglian Xiexintang showed significantly reduced body weight and epididymal fat weight (P<0.01). The epididymal fat index was reduced in all intervention groups (P<0.05), and FBG was lowered in all intervention groups (P<0.01).Serum GSP, GLU, TG, and LDL-C levels were reduced in the metformin group and the high- and medium-dose groups of modified Dahuang Huanglian Xiexintang (P<0.05, P<0.01). The serum TC level was significantly reduced in the metformin group and high-dose group of modified Dahuang Huanglian Xiexintang (P<0.01), and HDL-C levels were significantly increased in all intervention groups (P<0.05, P<0.01). The mRNA and protein expression of HIF-1α, BNIP3, and LC3B were significantly reduced, and UCP1 protein expression was significantly increased in the metformin group and high- and medium-dose groups of modified Dahuang Huanglian Xiexintang (P<0.05, P<0.01). The mRNA and protein expression of p62, Dio2, and Prdm16 were significantly increased in the metformin group and high-dose group of modified Dahuang Huanglian Xiexintang (P<0.05, P<0.01). ConclusionModified Dahuang Huanglian Xiexintang may inhibit mitochondrial autophagy and promote the browning of visceral adipose tissue through the HIF-1α/BNIP3/LC3B pathway, thereby improving glucose and lipid metabolism in obese T2DM rats.
4.Dahuang Huanglian Xiexintang and Its Modified Prescription Improve Type 2 Diabetes Mellitus: A Review
Dong AN ; Yanhui ZHAI ; Yankui GAO ; Rong LIU ; Qi ZHOU ; Xiangdong ZHU ; Yonglin LIANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(24):141-151
Type 2 diabetes mellitus (T2DM) is based on insulin resistance (IR) and insulin secretion deficiency, with the specific mechanisms still unclear. Current research involves mechanisms such as glycolipid toxicity, inflammatory response, oxidative stress damage, and mitochondrial dysfunction. Modern traditional Chinese medicine (TCM) scholars have named it "blood glucose collateral disease" based on the clinical characteristics and natural progression of T2DM. This condition is primarily manifested as abnormal blood sugar levels in the early stages, and as the disease progresses, it gradually causes widespread damage to the body's veins and collaterals, ultimately leading to lesions in vessels and collaterals. Among these, "spleen heat" (obesity type) is the most common clinical type of T2DM. The concept of "internal heat-induced elimination" runs through both the onset and complications of T2DM, with internal heat being a key factor in its pathogenesis. The clinical application of Dahuang Huanglian Xiexintang and its modifications has achieved significant therapeutic effects. This paper reviews the origins and treatment characteristics of Dahuang Huanglian Xiexintang, along with clinical application research and experimental studies related to T2DM treatment, involving mechanisms for regulating glucose and lipid metabolism disorders, improving IR, modulating inflammatory responses, combating oxidative stress damage, regulating autophagy-related signaling pathways, modulating intestinal flora, inhibiting pyroptosis, and alleviating endoplasmic reticulum stress, with the purpose to provide direction for further research on the prevention and treatment of T2DM and its related complications, to offer reference for developing Dahuang Huanglian Xiexintang as a rapid hypoglycemic Chinese patent medicine for obese T2DM, and to better guide the clinical promotion of this drug.
5.The effects of repeated high acceleration on implant osseointegration in SD rats
Xiaoni ZHOU ; Xiangdong LIU ; Yingbo JI ; Shuai HUANG ; Yingliang SONG ; Wei MA
Journal of Practical Stomatology 2024;40(2):204-209
Objective:To investigate the effects of repeated high acceleration(+Gz)on implant osseointegration in SD rats.Methods:18 SD rats were divided into+Gz and control groups randomly(n=9),and 1 implant was placed in each femur of the rat's lower limb.24 hours postoperatively,the experimental rats were exposed to+Gz of 4 to 9 G with 1 G/s environment 3 times a week,while the con-trol rats were fed normally.3 rats from each of the 2 groups were sacrificed at 2,4 and 8 weeks after implantation.Micro-CT,sequential fluorescence double labeling,and histological examination were perfomed for the analysis of implant osseointegration.Results:The bone volume fraction(BV/TV),trabecular number(Tb.N),mineral apposition rate(MAR),implant-bone contact rate(BIC)and bone area in implant thread(BA)of the+Gz group were significantly lower than those of the control group at 2 weeks(P<0.05),and so as to MAR,BA at 4 weeks(P<0.05),while there was no significant difference of the parameters at 8 weeks after implantation.Conclusion:In SD rats early exposure to+Gz environment postoperatively may have a negative effect on initial osseointegration by slowing bone forma-tion.However,it will not lead to poorer bone mass when sustained over a long period.
6.Mechanism Exploration of TG Regulating PI3K/Akt to Improve Insulin Resistance in Liver of T2DM Rats Based on Transcriptomics
Qin LI ; Yonglin LIANG ; Xiaowei SHI ; Xuan LIU ; Xiangdong ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(2):99-109
ObjectiveTo investigate the effect of Tangzhi pills on the improvement of insulin resistance (IR) in the liver with type 2 diabetes (T2DM) by regulating phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway based on differential genes and its possible molecular mechanism. MethodT2DM rat models were prepared by high fat (HFD) diet combined with streptozotocin (STZ) intraperitoneal injection. The experiment was divided into blank group, model group, metformin hydrochloride group (0.18 g·kg-1), Tangzhi pills high (1.08 g·kg-1), medium (0.54 g·kg-1) and low (0.27 g·kg-1) dose groups. Rat serum, liver, and pancreatic tissue were collected, and the pathological tissue of the liver and pancreas was observed using hematoxylin-eosin (HE) staining. The fasting blood glucose level (FBG) was detected, and oral glucose tolerance (OGTT) tests were conducted. Enzyme-linked immunosorbent assay (ELISA) was used to detect fasting serum insulin (FINS) and glycated hemoglobin (GHb) levels in rats. IR homeostasis model index (HOMA-IR), β cellular homeostasis index (HOMA-β), and insulin sensitivity index (ISI) were calculated. Biochemical methods were used to determine the levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL-C), and high-density lipoprotein (HDL-C) in rat serum. Transcriptomics obtained differentially expressed mRNA from liver tissue and enriched differentially expressed pathways. Real-time reverse transcriptase polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of cyclic adenylate responsive element binding protein 3-like protein 2 antibody (CREB3l2), B-lymphocyte tumor 2 (Bcl-2), Toll-like receptor 2 (TLR2), cyclin-dependent kinase inhibitor 1A (CDNK1A), and DNA damage induced transcription factor 4-like protein (DDIT4) in liver tissue. Western blot was used to detect the protein expression of phosphorylated phosphatidylinositol 3-kinase (p-PI3K), phosphorylated protein kinase B (p-Akt), glucose transporter 4 (GLUT4), insulin receptor (INSR), and insulin receptor substrate 2 (IRS2). ResultThe pharmacodynamic experiment results showed that compared with model group, Tangzhi pills groups repaired liver and pancreatic tissue to varying degrees, reduced blood sugar (P<0.01), and promoted a decrease in serum FINS, GHb, and HOMA-IR (P<0.05, P<0.01). In addition, HOMA-β and ISI increased (P<0.05, P<0.01). The levels of TC, TG, and LDL-C decreased (P<0.05, P<0.01), while the levels of HDL-C increased (P<0.05, P<0.01). The transcriptomics experimental results confirmed that the PI3K/Akt signaling pathway was significantly expressed in both the blank group and model group, as well as in the high-dose Tangzhi pills group and model group. CDNK1A, DDIT4, CREB3l2, Bcl-2, and TLR2 were significantly differentially expressed mRNA during TG intervention in T2DM. Compared with the model group, the protein expression of p-PI3K, p-Akt, GLUT4, INSR, and IRS2 increased in all Tangzhi pills groups (P<0.01). The mRNA expression of CREB3l2, Bcl-2, and TLR2 increased (P<0.01), while that of CDNK1A and DDIT4 decreased (P<0.01). ConclusionTangzhi pills may regulate the PI3K/Akt signaling pathway based on the differential mRNA expression of CREB3l2, Bcl-2, TLR2, CDNK1A, and DDIT4, thereby improving IR in the liver with T2DM.
7.Analysis of DMD gene variants in a single center
Chunxiao HUA ; Lina LIU ; Shaozhe YANG ; Shuwen XIN ; Xiangdong KONG
Chinese Journal of Pediatrics 2024;62(2):153-158
Objective:To investigate the DMD genetic variants of the Chinese population with Duchenne (DMD) and Becker muscular dystrophies (BMD).Methods:A cross-sectional study was conducted on 2 690 unrelated patients with DMD and BMD aged 0-18 who visited the Genetic and Prenatal Diagnosis Center of the First Affiliated Hospital of Zhengzhou University from January 2005 to February 2022. The clinical data, such as gender, age, clinical manifestations, and address, were collected. Multiplex ligation-dependent probe amplification, next generation sequencing panel, Sanger sequencing, and PCR amplification were used to detect the variants of the DMD gene in the patients, whose clinical information and gene detection results were descriptively analyzed.Results:The 2 690 patients included 2 648 males and 42 females, with an age of 6.0 (4.0, 9.0) years. The serum creatine kinase increased in all patients. Pathogenic DMD gene variants were detected in the 2 618 patients, including 1 875 cases (71.6%) large deletions, 231 cases (8.8%) duplications, and 512 cases (19.6%) small variants. Among the deletion variants, the deletion of 3 exons was the most common, accounting for 15.4% (288/1 875); and hotspot deletion involved exons 45 to 50, accounting for 6.3% (119/1 875). Exon 2 was the most common type duplication region, accounting for 13.0% (30/231). Small variants were distributed in all 79 exons of the DMD gene, with no hotspots. In addition, the 46 small variants were previously unreported.Conclusion:Exon deletion is the most common type of DMD gene variant, followed by small variants and exon duplication.
8.Effect of macrophages polarization on proliferation,migration and osteogenic differentiation of periodontal ligament stem cells
Kepeng LI ; Zhenguo SHEN ; Xiangdong LIU ; Tiantian CHENG ; Yuanyin WANG
Acta Universitatis Medicinalis Anhui 2024;59(8):1392-1398
Objective To explore the effects of different phenotypes macrophages(Mφs)on the proliferation,mi-gration and osteogenic differentiation of periodontal ligament stem cells(PDLSCs).Methods PDLSCs were isola-ted and cultured by tissue block method.Tohoku Hospital Pediatrics-1(THP-1)cell line was stimulated to activate into unpolarized Mφs(M0),then induced to polarize into type Ⅰ Mφs(M1)and type Ⅱ Mφs(M2).Quantitative real-time PCR(qPCR)detected the inflammatory factors tumor necrosis factor α(TNF-α),interleukin(IL)-1 β,IL-6,IL-10 and transforming growth factor-β(TGF-β)mRNA expression level.After collecting culture superna-tants with different phenotypes,PDLSCs were stimulated,native control(NC)group did not receive the culture su-pernatant of Mφs.The effects of PDLSCs proliferation were assessed via Methylthiazolyldiphenyl-tetrazolium bro-mide(MTT)assay,while scratch assays were employed to evaluate their migration.Western blot was utilized to analyze the protein expression of Runt-related transcription factor 2(RUNX2)and alkaline phosphatase(ALP).Additionally,Alizarin Red staining was performed to investigate the deposition of calcified nodules in PDLSCs.Re-sults qPCR showed the relative expression of TNF-α,IL-1 β and IL-6 in M1 Mφs were higher than those in M0 and M2 Mφs(P<0.05),and the relative expression of IL-10 and TGF-β in M2 Mφs were higher than those in M0 and M1 Mφs(P<0.05);Western blot showed the expression of RUNX2 and ALP proteins in PDLSCs in M0 and M2 groups was higher than those in the NC group(P<0.05),Alizarin Red staining showed increased calcified nodule deposition in PDLSCs in M0,M1 and M2 groups compared to the NC group;MTT assay showed the prolifer-ation of PDLSCs in the M0 and M1 groups was suppressed compared to the NC group(P<0.05);and scratch ex-periment showed the migratory capacity of PDLSCs in the M1 and M2 groups was stronger than that in the NC group.Conclusion M0 and M1 Mφs inhibit PDLSCs proliferation,M1 and M2 Mφs promote PDLSCs migration,and all types of Mφs promote osteogenic differentiation of PDLSCs.
9.Research progress of family resilience in patients with malignant tumors
Tingting LIU ; Xiangdong ZHOU ; Jing DENG
Chongqing Medicine 2024;53(22):3485-3489
Family resilience refers to the positive response of families in the face of adversity in order to promote family recovery and maintain family function and structural stability.This paper combs the related researches on family resilience in the patients with malignant tumors at home and abroad,introduces the con-cept of family resilience,evaluation tools,research status quo and influencing factors,and provides the corre-sponding intervention strategy for family resilience in the patients with malignant tumors in order to provide reference for optimizing more comprehensive and more effective service of the managers or medical institu-tions.
10.Sufu mutation promotes malignant progression of lung adenocarcinoma by regulating Gli1
Jihong LIU ; Hu LUO ; Xinglin YI ; Xiangdong ZHOU
Journal of Army Medical University 2024;46(10):1142-1149
Objective To explore the effect of Sufu on lung adenocarcinoma by observing its mutation and the changes in Gli1 transcription after its mutation.Methods cBioPortal for Cancer Genomics was used to obtain the mutation status of Sufu in lung cancer.Four Sufu single base mutation vectors were constructed and transfected into lung adenocarcinoma cell lines A549 and H1975 to establish cells with Sufu overexpression and knockout.Wounding healing assay was employed to determine the effect of Sufu on cell migration,and RT-qPCR,immunofluorescence assay and dual luciferase reporter gene assay were utilized to explore the changes in Gli1 transcription after Sufu mutation.Results cBioPortal data showed that the mutation rate of Sufu in lung cancer was 11.76%in squamous cell carcinoma(2/17),0.96%in adenocarcinoma(6/625),1.01%in small cell lung cancer(4/396),and 0.69%in non-small cell lung cancer(77/11 227).The median survival of lung cancer patients was significantly decreased in those with Sufu mutation than those without(38.00 vs 54.34 months,HR=1.943,P<0.05).A549 cells were sensitive to Sufu knockout,and it resulted in significant increase of the transcription level of Gli1(P<0.05).Overexpression of Sufu inhibited cell migration ability in both A549 and H1975 cells(P<0.05).Sufu mutation had no significant influence on the location of Gli1 in the cells.The Sufu-T411M mutation up-regulated the the transcription level of Gli1(P<0.05).Conclusion Sufu has an inhibitory effect on lung adenocarcinoma metastasis,and can be regarded as a potential target for lung adenocarcinoma metastasis.


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