1.Experimental study on effect of Weijinkang Oral Liquid on rats'models with pulmonary fibrosis caused by pneumonoconiosis
Tianxu GAO ; Dawen WEI ; Jiangyan XU ; Yuyao ZHAO ; Lei DU
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(03):-
Objective:To observe the effect of Weijinkang Oral Liquid on rats'models with siliconic nodules and the diffuse fibrosis of the lungs.Methods:45 rats were randomly divided into the blank group,dust-affected group,and treatment group.The rats in the blank group were fed in normal environment.The rats in the dust-affected group were given regular feed.The dust-affected rats in the treatment group were administrated 1.0mL/100g of Weijinkang condensed liquid in 30% by stomach perfusion,two times daily(33 times of human's dosage).Five of each group were killed and anatomized in turn on the 15th day,the 30th and the 90th day.Results:On the 90th day the pathological test showed massive cartilaginous changes projecting to the pulmonary surface in the dust-affected group.There was no abnormality of lungs in both treatment group and blank group.After receiving Weijinkang Oral Liquid the rats in the treatment group showed greater weights than those in the dust-affected group,and the weights of fresh and dehydrated lungs were lighter than those in dust-affected group.Conclusion:Weijinkang Oral Liquid had certain effect on siliconic nodules and the diffuse fibrosis of the lungs caused by pneumonoconiosis.
2.A novel 5' splice site mutation in ACTC1 gene may play an important role in congenital ventricular septal defect
Hang LI ; Binbin WANG ; Bingren GAO ; Jiangyan LIU ; Qiming ZHAO ; Wensheng CHEN ; Wei WANG ; Kun YANG
Chinese Journal of Tissue Engineering Research 2012;16(11):2087-2090
BACKGROUND: As a candidate gene of congenital heart disease, ACTC1 gene is related to congenital atrial septal defect inhumans.OBJECTIVE: To perform a mutation screen of ACTC1 gene in 110 nuclear families of congenital heart disease.METHODS: A case-control study was conducted based on 110 nuclear families of congenital heart disease and 300 normalhuman beings with no reported cardiac malformation. Six fragments in the coding region of ACTC1 gene was amplified by PCR invitro using five primers pairs. PCR products were screened for gene mutations.RESULTS AND CONCLUSION: A novel G-to-A variant was found at the third nucleotide of the intron downstream from exon 5.This mutation existed in a 5-year-old female with an isolated ventricular septal defect and her 30-year-old father, who had noreported cardiac anomalies. This mutation was not detected in 300 normal controls. These findings indicate that the mutation maybe related with congenital ventricular septal defects in humans.
3.Expression of interleukin-36 and its regulatory effect on monocyte function in patients with diabetic kidney disease
Jiangyan LI ; Yakun KONG ; Jianlin ZHAO ; Lichao ZHAO ; Chengyun XIONG ; Yanhong ZHOU
Chinese Journal of Endocrinology and Metabolism 2024;40(1):30-36
Objective:To analyze the level of interleukin-36(IL-36) family cytokines in peripheral blood, and explore the regulatory role of recombinant human IL-36α in monocyte function in patients with diabetic kidney disease(DKD).Methods:A total of 41 type 2 diabetes mellitus(T2DM) patients, 36 DKD patients, and 20 controls were consecutively enrolled. Plasma and peripheral blood mononuclear cells(PBMCs) were isolated. Enzyme-linked immunosorbent assay(ELISA) was used to measure plasma levels of IL-36α, IL-36β, IL-36γ, and IL-36 receptor antagonist(IL-36Ra). PBMCs were sorted, and real-time quantitative PCR was performed to detect the mRNA expression of IL-36 receptor subunits in monocytes. Monocytes were stimulated with recombinant IL-36α, and levels of cytotoxic molecules and cytokines in the culture supernatant were measured. Flow cytometry was used to assess the expressions of programmed death receptor-1(PD-1) and cytotoxic T-lymphocyte-associated protein 4(CTLA-4). Co-culture of monocytes with Vero cells was performed to evaluate monocyte cytotoxicity.Results:Plasma levels of IL-36α and IL-36β in the T2DM and DKD groups were significantly higher than those in the control group. The DKD group also showed higher plasma levels of IL-36α compared to the T2DM group( P<0.05). There were no significant differences in IL-36γ and IL-36Ra levels among the three groups( P>0.05). The mRNA expression of IL-36 receptor subunits in monocytes was comparable among the three groups( P>0.05). The DKD group had higher level of tumor necrosis factor-alpha(TNF-α) compared to the control and T2DM groups( P<0.05). The levels of PD-1 and CTLA-4 were lower in the DKD group than those in the control and T2DM groups( P<0.05). The proportion of monocyte-induced Vero cell death was significantly higher in the DKD group compared to the control and T2DM groups( P<0.05). After stimulation with recombinant human IL-36α, monocytes from DKD patients showed a significant increase in the secretion of granzyme B and TNF-α( P<0.05), as well as an increased proportion of monocyte-induced Vero cell death( P=0.024). Conclusion:In DKD patients, elevated IL-36α and granzyme B levels in monocytes enhance monocyte function.
4.Effects of recombinant human syntaxin-4(STX4) on lipopolysaccharide-induced islet β-cell INS-1 injury and underlying mechanism
Jianlin ZHAO ; Yufeng CHEN ; Suyu WU ; Jiangyan LI ; Chengyun XIONG ; Ying ZHANG ; Xiuqin GENG
Chinese Journal of Endocrinology and Metabolism 2021;37(8):738-744
Objective:To explore the effect of recombinant human syntaxin-4(STX4) on lipopolysaccharide(LPS)-induced injury in islet β-cells(INS-1).Methods:Pancreatic islet β-cells(INS-1) were divided into Control (blank control), LPS (LPS treatment), LPS+ NC (transfection of negative control vector, LPS treatment), and LPS+ STX4 (transfection of pcDNA-STX4, LPS treatment) groups. RT-qPCR and Western blot were used to detect STX4 mRNA and protein expression, flow cytometry to detect apoptosis, DCFHDA method to detect reactive oxygen species(ROS) level, xanthine oxidation method to detect superoxide orgotein dismutase(SOD) level, colorimetric method to detect glutathione peroxidase(GSH-Px) level, ammonium molybdate colorimetric method to detect catalase(CAT) level, thiobarbituric acid method to detect malonaldehyde(MDA) level, ELISA method to detect the level of interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and insulin secretion levels under glucose conditions secreted by cells, Western blot method to detect Cleared Caspase-3, Bcl-2 Associated X Protein(Bax), p65 protein expression. After treatment with NF-κB signaling pathway activator, STX4 up-regulated islet β-cell INS-1 was given LPS stimulation, and the same method was used to measure apoptosis, ROS, SOD, GSH-Px, CAT, MDA levels and secreted IL-1β, TNF-α, insulin levels and Cleaved Caspase-3, Bax, p65 protein levels.Results:Compared with the Control group, the expression of STX4 mRNA and protein in islet β cells of the LPS group decreased, the apoptosis rate, ROS level, and MDA levels increased, and the levels of SOD, GSH-Px, and CAT decreased, the levels of IL-1β, TNF-α increased, the level of insulin secreted by the cells decreased, and the expression levels of Cleaved Caspase-3, Bax, and p65 also increased. NF-κB pathway activator treatment reversed the effect of up-regulated STX4 on islet β-cell apoptosis, ROS, SOD, GSH-Px, CAT, MDA levels and secreted IL-1β, TNF-α levels, and Cleaved Caspase-3 , Bax and p65 protein levels.Conclusion:Up-regulation of STX4 alleviated LPS-induced islet β cell oxidative damage, apoptosis and inflammatory factor release. The underlying mechanism might be related to the inhibition of activated NF-κB signaling pathway.
5.Medication law of Traditional Chinese Medicine patent compounds in the treatment of Alzheimer disease based on data mining
Shanshan LI ; Min ZHAO ; Jiangyan SUN ; Jie ZHANG ; Yuanyuan YAN ; Zhihui CUI ; Menglong SHI
International Journal of Traditional Chinese Medicine 2023;45(2):220-226
Objective:To analyze the medication law of Traditional Chinese Medicine (TCM) patent compounds for Alzheimer disease (AD) by using data mining method.Methods:The TCM compounds for the treatment of AD in the patent database were screened, and the frequency, clustering and association analysis were carried out with the help of TCM inheritance calculation platform, SPSS Statistics 21.0 and SPSS Modeler 18.0 software. The medication law was analyzed.Results:A total of 220 patent compounds were included, involving 361 kinds of Chinese materia medica; the top 10 high-frequency drugs were Acori Tatarinowii Rhizoma, Polygalae Radix, Ginseng Radix et Rhizoma, Chuanxiong Rhizoma, Astragali Radix, Lycii Fructus, Poria, Rehmanniae Radix PraeparataAngelicae Sinensis Radix, Salviae Miltiorrhizae Radix et Rhizoma; the most frequently used drugs were drugs for tonifying deficiency and promoting blood circulation to remove blood stasis; most of their properties belonged to warm, mild and cold; the tastes were mainly sweet, bitter and pungent; the meridians belonged to the five internal organs. 16 items of association data (4 combinations of two items and 12 combinations of three items) were obtained by association rule analysis, and the strongest correlation group was " Acori Tatarinowii Rhizoma- Polygalae Radix" and " Acori Tatarinowii Rhizoma- Chuanxiong Rhizoma- Polygalae Radix". Cluster analysis showed four prescription combinations and three pairs of drug compatibility, including the addition and subtraction structure of Kaixin Powder, Zuogui Pill, Bazhen decoction and so on. Conclusion:The core treatment principle of TCM patent compound treatment of AD is regulating and tonifying the five internal organs to treat its root, resolving phlegm and removing blood stasis to treat the symptoms, which accords with the theoretical basis of TCM in the treatment of AD, and can provide reference for clinical practice and new drug research and development.
6.Clinical features of 59 patients with scrub typhus in Kunming City
Qin PENG ; Wu LI ; Yanghong ZOU ; Shenjun ZHAO ; Xiuling ZHANG ; Chunyan MOU ; Jiangyan ZHANG
Chinese Journal of Endemiology 2018;37(8):664-667
Objective To study the clinical features of patients with scrub typhus and provide scientific basis for clinical diagnosis and treatment. Methods Clinical data of patients with scrub typhus in the First Affiliated Hospital of Kunming Medical University from January 2016 to December 2016 were collected. Epidemiological data, clinical manifestations, laboratory findings, image examination results, treatment and outcome were retrospectively analyzed. Results The clinical manifestations included 59 cases (100.0%) with fever, 44 cases (74.6%) with headache, 39 cases (66.1%) with chills, 34 cases (57.6%) with muscle and joint pain, 29 cases (49.2%) with prostration, 49 cases (83.1%) with eschar or ulcer, 42 cases (71.2%) with lymphadenectasis, 23 cases (39.0%) with hepatosplenomegaly. Laboratory test results: 51 cases (86.4%) had normal or elevated white blood cell count, 50 cases of eosinophil reduced (84.7%), 27 cases of blood platelet (PLT) reduced (45.8%), 33 cases of albumin reduced (55.9%), 50 cases of alanine aminotransferase (ALT) increased (84.7%), 48 cases of aspartate aminotransferase (AST) increased (81.4%), and 56.1% (23/41) of the patients with triiodothyronine (T3), thyroxine (T4), free triiodothyronine (FT3) and free thyroxine (FT4) levels significantly lowered, with predominantly free FT3 reduction (82.6%,19/23); C reactive protein (CRP), procalcitonin (PCT), erythrocyte sedimentation rate (ESR) and ferritin were increased in 93.9%(46/49), 35.4% (17/48), 64.9% (24/37), and 83.8% (31/37) of the patients, and 95.6%(43/45) was accompanied with chest radiographic abnormalities. Tetracycline, doxycycline and azithromycin treatment were all effective. Conclusions The clinical manifestations of patients with scrub typhus, involving multisystem, are diverse and thyroid hormones decrease is observed. Early diagnosis and treatment is the key to improve the prognosis of patients with scrub typhus.
7. Research progress of dynamic regulation of lysine acetylation in heart disease
Yuan ZHAO ; Qianwen NIE ; Jiangyan YIN ; Xuan HUAN ; Meixiang TIAN ; Zhengyi ZHANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2021;26(10):1186-1192
Reversible post-translational modification of proteins is an important process in the physiological regulation of all tissues, including the heart. Lysine acetylation occurs in all organisms, including prokaryotes, and is regulated by a balance between lysine acetyltransferase (adding acetyl to the ε-amino group of lysine) and deacetylase (acetyl group that removes lysine ε-amino group). The heart is an organ rich in acetylated lysine, but the role of acetylated lysine in the heart remains to be elucidated. Therefore, in this paper, we systematically reviewed the gene list of acetyltransferase and deacetylase in mammalian genome and indicated their mRNA expression. The purpose of this study is to discover the research progress of dynamic regulation of lysine acetylation in heart disease and to provide a theoretical basis for the discovery of molecular targets.
8.Characterization of a monoclonal antibody against the hemagglutinin stem of H7N9 subtype avian influenza virus.
Jiangyan ZHAO ; Yanxiao ZHU ; Jiao HU ; Zenglei HU ; Xiufan LIU
Chinese Journal of Biotechnology 2022;38(1):160-173
The conserved hemagglutinin (HA) stem region of avian influenza virus (AIV) is an important target for designing broad-spectrum vaccines, therapeutic antibodies and diagnostic reagents. Previously, we obtained a monoclonal antibody (mAb) (5D3-1B5) which was reactive with the HA stem epitope (aa 428-452) of H7N9 subtype AIV. To systematically characterize the mAb, we determined the antibody titers, including the HA-binding IgG, hemagglutination-inhibition (HI) and virus neutralizing (VN) titers. In addition, the antigenic epitope recognized by the antibody as well as the sequence and structure of the antibody variable region (VR) were also determined. Moreover, we evaluated the cross-reactivity of the antibody with influenza virus strains of different subtypes. The results showed that the 5D3-1B5 antibody had undetectable HI and VN activities against H7N9 virus, whereas it exhibited strong reactivity with the HA protein. Using the peptide-based enzyme-linked immunosorbent assay and biopanning with a phage-displayed random peptide library, a motif with the core sequence (431W-433Y-437L) in the C-helix domain in the HA stem was identified as the epitope recognized by 5D3-1B5. Moreover, the mAb failed to react with the mutant H7N9 virus which contains mutations in the epitope. The VR of the antibody was sequenced and the complementarity determining regions in the VR of the light and heavy chains were determined. Structural modeling and molecular docking analysis of the VR verified specific binding between the antibody and the C-helix domain of the HA stem. Notably, 5D3-1B5 showed a broad cross-reactivity with influenza virus strains of different subtypes belonging to groups 1 and 2. In conclusion, 5D3-1B5 antibody is a promising candidate in terms of the development of broad-spectrum virus diagnostic reagents and therapeutic antibodies. Our findings also provided new information for understanding the epitope characteristics of the HA protein of H7N9 subtype AIV.
Animals
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Antibodies, Monoclonal
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Antibodies, Viral
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Hemagglutinin Glycoproteins, Influenza Virus/genetics*
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Hemagglutinins
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Influenza A Virus, H7N9 Subtype
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Influenza in Birds
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Molecular Docking Simulation
9.Shugan Jianpi Yangxin Prescription Regulates Orexin-A and Its Receptors in Mouse Model of Insomnia
Zhen WANG ; Jie ZHANG ; Feixiang LIU ; Jiangyan SUN ; Yahui GAO ; Yuxin ZHENG ; Min ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):54-61
ObjectiveTo investigate the interventional effects of Shugan Jianpi Yangxin prescription on the expression of orexin-A (OXA), orexin-1 receptor (OX1R), and orexin-2 receptor (OX2R) in the mouse model of insomnia. MethodThe mouse model of insomnia was established by intraperitoneal injection of DL-4-chlorophenylalanine (PCPA). Fifty BALB/c mice were randomized into a blank group, a model group, an eszopiclone (0.13 mg·kg-1) group, and low- and high-dose (8.4 and 33.6 g·kg-1, respectively) Shugan Jianpi Yangxin prescription groups and treated with the corresponding drugs for 14 consecutive days. The weight changes of mice were monitored, and Morris water maze and pentobarbital-induced sleep tests were conducted. Immunohistochemistry (IHC) was employed to examine the expression of OXA in the hypothalamus. Enzyme-linked immunosorbent assay was used to measure the levels of OXA and 5-hydroxytryptamine (5-HT) in the hypothalamus, serum, and spleen. Real-time fluorescence quantitative polymerase chain reaction was employed to determine the mRNA levels of OXA, OX1R, and OX2R in the hypothalamus. ResultCompared with the blank group, the model group had decreased body weight (P<0.01), increased escape latency (P<0.01), increased sleep latency (P<0.01), shortened sleep duration (P<0.01), elevated OXA level and lowered 5-HT level in the hypothalamus, serum, and spleen (P<0.05), and up-regulated mRNA levels of OXA, OX1R, and OX2R in the hypothalamus (P<0.01). Compared with the model group, the low- and high-dose groups of Shugan Jianpi Yangxin prescription showed increased body weight (P<0.05, P<0.01), shortened escape latency (P<0.05), shortened sleep latency and prolonged sleep duration (P<0.01), and lowered OXA level and elevated 5-HT level in the hypothalamus, serum, and spleen (P<0.05, P<0.01). Moreover, the two doses of Shugan Jianpi Yangxin prescription down-regulated the mRNA levels of OXA, OX1R, and OX2R in the hypothalamus (P<0.01). ConclusionShugan Jianpi Yangxin prescription exerts sedative and hypnotic effects in mice by increasing the content of 5-HT in the brain and inhibiting the expression of OXA and its receptors in the hypothalamus.
10.Mechanism of Yishen Tongluo Prescription in Inhibiting Endoplasmic Reticulum Stress and Improving Apoptosis of Renal Tubular Epithelial Cells Based on PERK/ATF4/CHOP
Xuan SU ; Liang ZHAO ; Mengmeng WANG ; Jing DING ; Zhenqiang ZHANG ; Xiaowei ZHANG ; Jiangyan XU ; Zhishen XIE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(2):26-36
ObjectiveTo investigate the effect of Yishen Tongluo prescription (YSTLP) on apoptosis of renal tubular epithelial cells and explore the mechanism based on endoplasmic reticulum stress pathway of protein kinase R-like endoplasmic reticulum kinase (PERK)/activating transcription factor 4 (ATF4)/transcription factor C/EBP homologous protein (CHOP). MethodThe db/db mice were randomly divided into model group, valsartan group (10 mg·kg-1), and low, middle, high-dose YSTLP groups (1, 2.5, 5 g·kg-1). Samples were collected after eight weeks of drug intervention. In addition, db/m mice in the same litter served as the control group. Human renal tubular epithelial cells (HK-2) were cultured in vitro and divided into the control group, advanced glycated end-product (AGE) group, and AGE + low, middle, and high-dose YSTLP groups (100, 200, 400 mg·L-1). TdT-mediated dUTP nick end labeling (TUNEL) staining was used to detect the apoptosis rate of HK-2 cells. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay was conducted to detect the viability of HK-2 cells. Calcium fluorescence probe staining and luciferase reporter gene method were adopted to detect the luciferase activity of folded protein response element (UPRE) and endoplasmic reticulum stress. Immunohistochemical (IHC) analysis was carried out to measure the protein expressions of phosphorylated PKR (p-PERK), CHOP, and ATF4. Real-time polymerase chain reaction (Real-time PCR) was used to measure the mRNA expression levels of CHOP and X-box binding protein 1 (XBP1) in mouse kidney and HK-2 cells. Western blot was used to detect the protein expression level of p-PERK, PERK, CHOP, ATF4, B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), and cleaved Caspase-3 in mouse kidney and HK-2 cells. ResultIn the cellular assay, HK-2 cell viability was significantly reduced, and the apoptosis rate was elevated in the AGE group compared with the control group (P<0.01). The mRNA and protein expression levels of apoptosis-related factor Bcl-2 were significantly reduced (P<0.01), and those of Bax were significantly increased (P<0.01). The protein expression level of cleaved Caspase-3 was significantly increased (P<0.01). Compared with the AGE group, YSTLP administration treatment resulted in elevated cell viability and reduced apoptosis rate (P<0.01). The mRNA and protein expression levels of Bcl-2 were significantly elevated in a time- and dose-dependent manner (P<0.01), and those of Bax were significantly reduced in a time- and dose-dependent manner. The protein expression level of cleaved Caspase-3 was significantly reduced in a time- and dose-dependent manner (P<0.01). The intracellular Ca2+ imbalance and UPRE luciferase fluorescence intensity were increased in the AGE group compared with the control group (P<0.01). The mRNA levels of endoplasmic reticulum stress-related factors CHOP and XBP1 were significantly increased (P<0.01), and the protein expression levels of p-PERK, CHOP, and ATF4 were significantly increased (P<0.05). Compared with the AGE group, YSTLP effectively improved intracellular Ca2+ imbalance in HK-2 cells and decreased UPRE luciferase fluorescence intensity in a dose-dependent manner (P<0.01). It reduced the mRNA levels of endoplasmic reticulum stress-related factors CHOP and XBP1 (P<0.01) and the protein expression levels of intracellular p-PERK, CHOP, and ATF4 in a dose- and time-dependent manner (P<0.01). In animal experiments, the protein expression level of Bcl-2 was significantly reduced(P<0.01), and that of cleaved Caspase-3 and Bax was significantly increased in the model group compared with the control group (P<0.05). The protein expression level of Bcl-2 was dose-dependently elevated, and that of cleaved Caspase-3 and Bax was dose-dependently decreased in the YSTLP groups compared with the model group (P<0.01). Compared with the control group, the mRNA expression levels of CHOP and XBP1 were significantly elevated in the model group (P<0.05, P<0.01), and the protein expression levels of p-PERK, CHOP, and ATF4 were significantly increased (P<0.05). Compared with the model group, YSTLP significantly decreased the mRNA expression levels of CHOP and XBP1 (P<0.01) and the protein expression levels of p-PERK, CHOP, and ATF4 (P<0.01). ConclusionYSTLP can effectively inhibit endoplasmic reticulum stress and improve apoptosis of renal tubular epithelial cells, and its mechanism may be related to the regulation of the PERK/AFT4/CHOP pathway.