1.The study of the correlation between serum C-reactive proteln,tumor necrosis factor-alpha and atherosclerosis in patients with subclinnical hypothyroidism
Jianping GONG ; Shuping FU ; Juan YIN ; Jin XIE ; Xue LIAN ; Yaqing GUO
Chinese Journal of Postgraduates of Medicine 2012;35(16):14-17
ObjectiveTo investigate the correlation between serum C-reactive protein(CRP),tumor necrosis factor-alpha(TNF-α) and atherosclerosis in patients with subclinical hypothyroidism.Methods Seventy-five patients with subclinical hypothyroidism were divided into two groups according to the level of TSH:mild group (42 cases) with TSH between 5.5 and 10.0 mU/L and severe group (33 cases) with TSH >10.0 mU/L.Thirty healthy physical examinees or volunteers were selected as control group.TG,LDL-C,HDL-C,TSH,FT4,FT3,CRP,TNF-α and carotid intima-media thickness(CIMT) of all cases were detected and measured.ResultsCIMT of mild,severe and control groups were(0.88 ± 0.20),( 1.12 ±0.21 ),(0.62 ±0.21 ) mm,respectively.CIMT of mild and severe groups were both significantly higher than that of control group,CIMT of severe group was significantly higher than that of mild group (P < 0.01 ).There were no significant differences of CRP and TNF- α among three groups (P > 0.05 ).LDL-C of mild and severe groups were significantly higher than that of control group [ (3.22 ± 0.37),(3.49 ± 0.38 ) mmol/L vs.(2.48 ±0.41 )mmol/L,P <0.01 ].HDL-C and TG between mild and control groups had no statistical significance (P >0.05).Compared with control and mild groups,HDL-C of severe group decreased [(0.92 ±0.10)mmol/L vs.( 1.21 ± 0.14),( 1.17 ± 0.11 ) mmol/L] and TG increased [ ( 1.50 ± 0.49) mmol/L vs.( 1.11 ±0.53),(1.27 ±0.47) mmol/L](P< 0.01 or < 0.05).The levels of TG,LDL-C,TSH,CRP and TNF-α had positive correlation with CIMT (r =0.52,0.37,0.48,0.39,0.45;P <0.05 or <0.01 ).FT4 was negatively correlated with CIMT (r=-0.24,P<0.05).HDL-C had no relation with CIMT (r =0.06,P >0.05).ConclusionsThere are many abnormalities such as metabolism disorder of blood lipid in patients with subclinical hypothyroidism.The risk of atherosclerosis increases in these patients and inflammatory reaction may not be an important factor of subclinical hypothyroidism complicated with atherosclerosis.
2.PCR-HRM Analysis for Gene Mutation Screening in a Child with Osteogenesis Imperfecta
Xue BAI ; Keqiu LI ; Xiuzhi REN ; Xiaobo HE ; Yi WANG ; Shizhen GUAN ; Yaqing JING ; Guang LI
Tianjin Medical Journal 2014;(7):693-696
Objective To investigate COL1A1 gene mutation by PCR-high resolution melting (PCR-HRM) and an-alyze the correlation between genotype and clinical phenotype in a child (proband) with osteogenesis imperfecta (OI). Methods The family history of OI pedigree along with the clinical data was collected. Blood samples from the proband and his family members, as well as 50 normal controls, were collected. The mutation of COL1A1 gene was screened using PCR-HRM and validated by the gene sequence. Results The detection of PCR-HRM showed the abnormal result of COL1A1 17 exon in proband with a lower melting temperature (Tm) value than that of normal controls by 0.4℃. There were signifi-cant differences in the standardization melting curve and the different melting curve between the proband and the normal controls. The sequencing result was c.1138G>A, which meant that cDNA of 1138 base G mutation into A. The mutations transformed the amino acid glycine into a serine at amino acid 380(P. Gly 380 Ser), which resulted in missense mutations. The proband’s father and grandmother had the same mutation of COL1A1 gene. The mutation was not found in the proband’s mother and normal controls. There was no report for such mutation in Chinese population. Pedigree analysis showed the fami-ly genetic characteristics of autosomal dominant inheritance. The proband was clinically diagnosed as OI type Ⅳwith more severe clinical phenotype. Conclusion PCR-HRM analysis is a new effective method for genetic screening of OI. COL1A1 mutation of c.1138G>A is a newly discovered mutation in Chinese population. Gly replaced inαhelical domain may lead to a more severe clinical phenotype.
3.Screening and analysis of a new mutation of COL1A1 gene in a family with osteogenesis imperfecta.
Xue BAI ; Keqiu LI ; Xiuzhi REN ; Xiaobo HE ; Yi WANG ; Shizhen GUAN ; Yaqing JING ; Guang LI
Chinese Journal of Medical Genetics 2014;31(3):344-347
OBJECTIVETo investigate mutation of COL1A1 gene and analyze the relationship between genotype and clinical phenotype in a family with osteogenesis imperfecta (OI).
METHODSThe family history of an OI pedigree, along with clinical data, was collected. Blood samples from the proband and his families, as well as 50 normal controls, were collected. Mutation of COL1A1 gene was screened using PCR-high resolution melting (PCR-HRM) and validated by sequencing.
RESULTSPCR HRM method showed an abnormal result in proband COL1A133_34 exons, which Tm was 87.7℃, in contrast to the normal control (wt) Tm of 87.9±0.06℃. There was a significant difference between the proband and the normal control with the standardization curve and the difference curves. DNA sequencing showed that Y9COL1A1 gene exons 33_34 has lost a C base (c.2321delC), which resulted in a frameshift mutation and caused an premature termination codon (UAA) at amino acid 334, i.e., p.Pro774LeufsX334 The father and grandfather of the proband, both suffered from OI, were verified to be heterozygous for the same mutation. The same mutation was not found in 50 normal controls. Database search confirmed this to be a novel mutation. Pedigree analysis suggested that it has an autosomal dominant inheritance. The proband and patients from the family were clinically diagnosed as OI type I.
CONCLUSIONThe study has identified a novel mutation of COL1A1 gene, c.2321delC. This frameshift mutation has caused a premature stop codon and reduced collagen type synthesis, characterized by a lighter OI clinical phenotype.
Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child ; Child, Preschool ; China ; Collagen Type I ; genetics ; Female ; Frameshift Mutation ; Humans ; Male ; Molecular Sequence Data ; Osteogenesis Imperfecta ; genetics ; Pedigree
4.Research status and evolution of health management in China from 2011 to 2020
Chichen ZHANG ; Xiao ZHENG ; Yaqing XUE ; Lei SHI ; Yi QIAN ; Ping OUYANG ; Hong ZHU ; Minsheng CHEN
Chinese Journal of Health Management 2021;15(6):567-573
Objective:To analyze research status and development trends in the field of health management in China from 2011 to 2020.Methods:“CNKI” was chosen as the data source, and “health management(precise)” was used as the search term, and a total of 13, 686 valid data were finally obtained. Frequency counts were used to tabulate the number of articles published in the field of health management from 2011 to 2020. CiteSpace software was used to analyze the cooperation of institutions, and to explore the research hotspots and development trends in the field of health management by institutions co-occurrence, keyword co-occurrence and clustering timeline map. Bicomb software and SPSS 26.0 software were used for multi-dimensional scale analysis of keywords to comprehensively reflect the core degree and maturity of research topics.Results:The amount of domestic health management research literature had shown an increasing trend from 2011 (804) to 2020 (2 044). The top 5 keywords in terms of frequency were “hypertension(611)” “diabetes(577)” “health education(485)” “community(460)” and “chronic diseases(457)”. “Elderly” “Traditional Chinese Medicine(TCM health management)” and “Health management model” were the hot keywords and research trends of health management. There were 7 themes in the field of health management, namely “Construction and application of chronic diseases health management model” “Community health service and health management” “Health management in essential public health service” “Health management of the elderly” “Health management of Traditional Chinese Medicine” “Health examination and health management organization” “Health management based on big data and modern information technology”.Conclusions:A relatively close network of cooperation has been formed in the field of health management research and the number of articles has increased. The elderly, chronic disease and Traditional Chinese Medicine health management are the research trend. The construction and implementation of health management models, the integration of artificial intelligence and health management are the development trends in this field.
5.Optimization of the azomethane oxide and dextran sodium sulfate model of colitis-associated colon cancer and changes in the intestinal microbiota
Dunfang WANG ; Lin ZHU ; Xue FENG ; Caijuan ZHANG ; Haifan LIU ; Yaqing LIU ; Bin LIU ; Li LIU ; Weipeng YANG
Acta Laboratorium Animalis Scientia Sinica 2024;32(2):151-160
Objective To optimize the method of combining azomethane oxide(AOM)and dextran sodium sulfate(DSS)to create a colitis-associated colon cancer(CAC)model,and to explore the pathogenesis of the intestinal flora in CAC.Methods Model groups A and B were established by one and two injections of AOM,respectively,combined with free drinking of DSS,and a normal control group was injected intraperitoneally with normal saline combined with purified water(n=10 mice per group).The better modeling scheme was selected by comprehensive evaluation of the disease activity index score,colon length,tumor rate,and mortality.Serum levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and tumor markers CA199,CEA,and CA724 were detected by enzyme-linked immunosorbent assay.Colon lesions were evaluated by hematoxylin and eosin(HE)staining.Changes in the intestinal microbiota in CAC mice were detected by 16S rDNA high-throughput gene sequencing analysis of mouse feces.Results Both single and enhanced AOM injections combined with DSS induced CAC mice;however,colon growths were larger,more closely arranged,and their morphological size was more consistent in group B compared with group A,with a tumor-formation rate of 100%.IL-6 levels were increased in the model group compared with the normal group(P<0.05).TNF-α levels were increased in the model group compared with the normal group(P>0.05).The CA199 and CEA levels were also significantly increased(P<0.05),but CA724 levels were not.Infiltration of inflammatory cells in the colon detected by HE pathology was accompanied by high-grade intraepithelial tumor-like changes on the surface of the lumen.The diversity and abundance of intestinal bacteria were decreased in CAC mice compared with normal mice:phyla Verrucomicrobiota and Actinobacteriota were significantly increased(P<0.05),Bacteroidota and Campilobacterota were significantly decreased(P<0.05).Akkermansia,Prevotellaceae,Ruminococcus,and Bifidobacterium were significantly increased(P<0.05),and Roseburia,Rikenellaceae_RC9_gut_group,Anaeroplasma,and Muribaculaceae were significantly decreased(P<0.05).Conclusions Two injections of AOM combined with 1.5%(1.5 g/100 mL)DSS induced CAC model mice with a high colon-tumorigenesis rate,uniform tumor morphology,and low mortality,and may thus be the preferred modeling scheme for pharmacodynamic experiments.Disorders or dysfunction of the intestinal flora may lead to increased permeability,loss of intestinal mucosal barrier function,and the release of enterogenic endotoxins,Resultsing in a sustained inflammatory response,as an indirect or direct cause of CAC pathogenesis.
6.Mechanism of Huangqintang in Treatment of Ulcerative Colitis and Related Colon Cancer: A Review
Xue FENG ; Yaqing LIU ; Bin LIU ; Xuran MA ; Dunfang WANG ; Weipeng YANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(7):1-10
Ulcerative colitis (UC) is a chronic intestinal disease with unknown etiology, with main symptoms of abdominal pain, diarrhea, mucus, pus, and blood in the stool. It can be accompanied by various complications and has a high risk of developing to colon cancer. In recent years, the incidence of UC and related colon cancer has been increasing, which seriously affects human health and quality of life. The operation, immunosuppressant, etc. are the main approaches in the modern clinical treatment of UC and related colon cancer, but these methods all have different toxic and side effects, and the therapeutic effect is not ideal. For many years, traditional Chinese medicine (TCM) has attracted much attention in the treatment of UC and related colon cancer due to its slightly toxic side effects and remarkable curative efficacy. Huangqintang, derived from the Shang Han Lun (伤寒论), is composed of Scutellariae Radix, Paeoniae Radix Alba, Glycyrrhizae Radix et Rhizoma, and Jujubae Fructus with the functions of clearing heat, checking diarrhea, harmonizing the middle, and relieving pain, and has a significant effect on the treatment of UC. Huangqintang has complex compositions and plays roles with multiple targets and pathways. According to the literature and the research results of this research group for many years, it was found that the mechanism of Huangqintang in the treatment of UC and related colon cancer was presumably related to the protection of the intestinal mucosal barrier, inhibition of inflammatory response, promotion of mitophagy, inhibition of oxidative stress, regulation of intestinal flora, cell cycle, and gene expression, suppression of cell proliferation, and promotion of apoptosis. To provide theoretical references for an in-depth study of the mechanism and clinical use of Huangqintang, this paper reviewed the research advances in recent years.
7.Effect of Huangqintang on Intestinal Flora in Mice with Ulcerative Colitis and Its Protective Mechanism on Intestinal Mucosal Barrier
Yaqing LIU ; Hangyu XU ; Dunfang WANG ; Bin LIU ; Xuran MA ; Xue FENG ; Weipeng YANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(7):11-19
ObjectiveTo evaluate the pharmacodynamic effect of Huangqintang (HQT) on ulcerative colitis (UC) model mice and investigate its protective effect against UC by regulating intestinal flora. MethodMale Balb/c mice were randomly divided into control group,model group, high-, medium-, and low-dose HQT groups (20, 10, 5 g·kg-1), flora interference group, flora interference model group, and flora interference-drug treatment group (HQT, 20 g·kg-1). The flora interference model was constructed through intragastric administration of antibiotics (200 mg·kg-1 bacitracin and 200 mg·kg-1 vancomycin) for 8 d, and the UC model was constructed by allowing mice with free access to 3% dextran sulfate sodium (DSS) solution for 7 d. HQT was administered for 7 d. After the experiments, the mice were sacrificed, and blood, colon, and feces were collected. Hematoxylin-eosin (HE) staining was performed to observe the colonic lesions. The serum levels of interleukin (IL)-4, IL-6, IL-10, and tumor necrosis factor (TNF)-α were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein expression of Claudin1, MUC1, Occludin, and zonula occludens-1(ZO-1) in colon tissues was detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. The fecal DNA of mice was extracted and analyzed by high-throughput sequencing. ResultCompared with the normal group, the model group showed increased serum content of IL-4, IL-6, and TNF-α (P<0.05, P<0.01) and decreased IL-10 (P<0.05). Compared with the model group, the HQT groups displayed decreased serum levels of IL-4, IL-6, and TNF-α (P<0.05, P<0.01), increased IL-10 content (P<0.01), increased mRNA and protein expression levels of Claudin1, MUC1, Occludin, and ZO-1 (P<0.05, P<0.01). After flora interference, the diversity and abundance of intestinal bacteria decreased. To be specific, Proteobacteria increased (P<0.01), and Firmicutes and Bacteroidetes decreased (P<0.01). After UC induction by DSS, Bacteroidetes and Tenericutes decreased (P<0.05). The high-, medium-, and low-dose HQT groups showed increased Bacteroidetes and Tenericutes (P<0.05, P<0.01) and decreased Firmicutes (P<0.05). Additionally, the abundance of Lactobacillus, Lachnospiraceae NK4A136 group, Escherichia-Shigella, and Helicobacteris was positively proportional to the dose of HQT. ConclusionHQT can inhibit the inflammatory response of UC mice, restore the imbalance of intestinal flora, and repair the damaged intestinal mucosal barrier.
8.Anti-inflammatory Effect of Huangqintang on LPS-induced RAW264.7 Inflammatory Cells
Yaqing LIU ; Bin LIU ; Xuran MA ; Dunfang WANG ; Xue FENG ; Weipeng YANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(7):20-28
ObjectiveTo explore the anti-inflammatory mechanism of Huangqintang based on the inflammation model in RAW264.7 cells. MethodHuangqintang was prepared and the safe dose to RAW264.7 cells was screened out. The RAW264.7 cells were seeded in 24-well plates and incubated with Huangqintang and lipopolysaccharide (LPS), successively. The concentrations of nitric oxide (NO), interleukin (IL)-6, tumor necrosis factor (TNF)-α, and prostaglandin E2 (PGE2) were measured by Griess assay and enzyme-linked immunosorbent assay (ELISA), respectively. Meanwhile, RAW264.7 cells were inoculated in 6-well plates, and normal group, LPS group, LPS+Huangqintang group, nuclear factor-κB (NF-κB) p65 inhibitor PDTC group, p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 group, extracellular signal-regulated kinase (ERK) inhibitor PD98059 group, c-Jun N-terminal kinase (JNK) inhibitor SP600125 group, and Janus kinase (JAK) inhibitor AG490 group were set up. After the cells were incubated with corresponding inhibitors and Huangqintang and stimulated by LPS, RNA and protein were extracted. The mRNA and protein expression levels of NF-κB p65, p38 MAPK, ERK, JNK, and JAK were detected by Real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively, to explore the anti-inflammatory mechanism of Huangqintang by regulating the NF-κB, MAPK, and JAK/signal transducer and activator of transcription protein (STAT) signaling pathways. ResultAfter stimulation with LPS, the concentrations of NO, IL-6, TNF-α, and PGE2 in the cells of the model group increased significantly(P<0.05,P<0.01). Compare with the model group, after incubation with Huangqintang, the secretion of NO, IL-6, TNF-α, and PGE2 showed a downward trend (P<0.05,P<0.01). Compared with the normal group, the model group showed increased mRNA expression of p38 MAPK, ERK, JNK, JAK, and NF-κB p65 and total protein expression in cells after stimulation with LPS (P<0.05,P<0.01). Compare with the model group,after incubation with Huangqintang, the total protein and mRNA expression of p38 MAPK, ERK, JNK, JAK, and NF-κB p65 in inflammatory cells decreased (P<0.05,P<0.01). Meanwhile, the expression of NF-κB p65 total protein and mRNA in each inhibitor group showed a downward trend (P<0.05,P<0.01). ConclusionHuangqintang can inhibit the inflammatory response through the NF-κB, MAPK, and JAK-STAT signaling pathways.
9.Anti-oxidative Stress Effect and Mechanism of Huangqintang on Caco-2 Cells Through Nrf2 Signaling Pathway
Xue FENG ; Yaqing LIU ; Bin LIU ; Xuran MA ; Dunfang WANG ; Weipeng YANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(7):29-37
ObjectiveTo verify the anti-oxidative stress effect of Huangqintang based on the nuclear factor E2-related factor 2 (Nrf2) signaling pathway by using Caco-2 cells as a carrier and RNA interference (RNAi) technology with in vitro experiments. MethodThe Caco-2 cells in the logarithmic growth phase were transfected with siRNA to construct siRNA Caco-2 cells. After normal Caco-2 cells and siRNA Caco-2 cells were incubated with Huangqintang of different doses, RNA and protein were extracted. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to detect the mRNA and protein expression of heme oxygenase-1 (HO-1), NAD(P)H quinone oxidoreductase 1 (NQO1), glutathione S-transferase (GST), Kelch-like ECH-associated protein 1 (Keap1), and Nrf2. Meanwhile, the activities of superoxide dismutase (SOD) and GSH-Px, as well as the expression levels of malondialdehyde (MDA) and reactive oxygen species (ROS), were detected by the colorimetric method and the probe method. ResultCompared with the results in the normal group, only the 400 mg·L-1 Huangqintang group and the sulforaphane (SFN) group could reduce the content of ROS and MDA in Caco-2 cells (P<0.01), while the activities of SOD and GSH-Px in the cells of the Huangqintang groups and the SFN group showed an upward trend. Furthermore, there were significant differences in the 400 mg·L-1 Huangqintang group/the SFN group and the normal group (P<0.01). Meanwhile, the protein and mRNA expression levels of HO-1, GST, Keap1, NQO1, and Nrf2 showed an upward trend in all groups (P<0.05, P<0.01). After transfection, compared with the normal group, the model group showed increased content of MDA and ROS, blunted activities of GSH-Px and SOD, and reduced protein and mRNA expression of HO-1, GST, Keap1, and NQO1 (P<0.05, P<0.01). After drug incubation, compared with the model group, the SFN group showed potentiated SOD activity, and the SFN group and the Huangqintang groups showed enhanced GSH-Px activity (P<0.01). Moreover, the activities of SOD and GSH-Px in the 400 and 200 mg·L-1 Huangqintang groups and the SFN group showed an upward trend (P<0.01), and the content of MDA in the 400 mg·L-1 Huangqintang group and the SFN group showed a downward trend. ROS decreased in all groups with drug intervention (P<0.01), and the protein and mRNA expression of HO-1, GST, Keap1, NQO1, and Nrf2 increased to varying degrees (P<0.05, P<0.01). ConclusionHuangqintang can play an anti-oxidative stress role by regulating the Nrf2 pathway.
10.Effect of Huangqintang on Th17/Treg and Th1/Th2 Balance in Mice with Ulcerative Colitis
Bin LIU ; Yaqing LIU ; Hongxin SONG ; Xuran MA ; Dunfang WANG ; Xue FENG ; Weipeng YANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(22):7-15
ObjectiveTo study the effect of Huangqintang on intestinal T helper cells(Th)17/regulatory T cells(Treg) and Th1/Th2 balance in experimental ulcerative colitis (UC) mice. MethodSixty male C57BL/6J mice were randomly divided into normal group, model group, sulfasalazine group (0.2 g·kg-1), and Huangqintang low-dose, medium-dose and high-dose groups (4.55,9.1,18.2 g·kg-1, respectively) according to body weight. Except for the normal group, all mice were given 2.5% dextran sodium sulfate (DSS, added to free drinking water) for 5 days to induce UC. After 7 days of continuous intragastric administration, the mice were sacrificed and the mesenteric lymph nodes (MLNs) and colon tissues were collected. The Th17/Treg and Th1/Th2 cell ratios in MLNs of mice were measured by flow cytometry. Interleukin-2 (IL-2), interleukin-17A (IL-17A), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-10 (IL-10) and other cytokines in colon tissues were detected by flow cytometric microsphere-based assay. The colon tissues samples were sectioned, and immunohistochemical method was used to detect the expression levels of T-box transcription factor (T-bet), GATA binding protein 3 (GATA-3), IL-17A and forkhead box P3 (FoxP3) in colon tissues. The content of transforming growth factor-β (TGF-β) in colon tissue of mice was determined by enzyme-linked immunosorbent assay (ELISA). ResultCompared with the normal group, the number of Th1 and Th17 cells in the model group was significantly increased, while the number of Th2 and Treg cells was significantly decreased, and Th1/Th2 and Th17/Treg were significantly increased (P<0.05, P<0.01). The expressions of T-bet, GATA-3, IL-17A and FoxP3 in colon tissues of mice were significantly increased (P<0.05, P<0.01), and the contents of cytokines IL-2, IL-17A, IL-6, TNF-α and IFN-γ were significantly increased. The contents of IL-4, TGF-β and IL-10 were significantly decreased (P<0.05, P<0.01). Compared with the model group, the imbalance of Th1/Th2 and Th17/Treg cells was reversed in each dose group of Huangqin decoction (P<0.01). The expressions of T-bet and GATA-3 in colon of mice were significantly decreased in low-dose, medium-dose and high-dose groups of Huangqintang (P<0.05, P<0.01), and the expression of IL-17A was significantly decreased in high-dose and low-dose groups of Huangqintang (P<0.01). The contents of cytokines IL-2, IL-17A, IFN-γ, IL-6 and TNF-α were significantly decreased in the high-dose and medium-dose groups of Huangqintang, and the contents of TGF-β and IL-10 were significantly increased in the high-dose group of Huangqintang (P<0.05, P<0.01). ConclusionHuangqintang could reduce colonic inflammation and improve colonic tissue damage in UC mice, possibly by regulating Th17/Treg and Th1/Th2 balance and related cytokines levels, and the release level of pro-inflammatory cytokines.