1.Intervention of Huayu Qutan Recipe on liver SREBP-2 signal pathway of hyperlipidemia rats of pi deficiency syndrome.
Xue LENG ; Lian-Qun JIA ; Guan-Lin YANG ; Jun-Yan WANG ; Mei-Lin ZHU ; Ying WANG ; Ying XU
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(3):320-326
OBJECTIVETo explore the intervention of Huayu Qutan Recipe (HQR) on liver SREBP-2 signal pathway of hyperlipidemia rats of Pi deficiency syndrome (PDS).
METHODSTotally 100 SPF grade SD rats were randomly divided into the blank control group, the hyperlipidemia group, the hyperlipidemia treatment group, the PDS hyperlipidemia group, and the PDS hyperlipidemia treatment group, 20 in each group. Common granular forage was fed to rats in the blank control group. High fat forage was fed to rats in the hyperlipidemia group and the hyperlipidemia treatment group. Rats in the PDS hyperlipidemia group and the PDS hyperlipidemia treatment group were treated with excessive labor and improper diet for modeling. They were administered refined lard by gastrogavage (3 mL each time, twice per day) and fed with high fat forage on the odd days, and fed with wild cabbage freely on even days. The modeling lasted for 30 days. Rats in the hyperlipidemia treatment group and PDS hyperlipidemia treatment group were administered with Huayu Qutan Recipe (20 mL/kg) by gastrogavage, once a day, for 30 successive days. Levels of serum cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), and serum amylase (AMY) were detected by automatic biochemical analyzer. D-xylose excretion rate was determined using phloroglucinol method. Morphological changes of liver and the lipid deposition in liver were observed using HE stain and oil red O stain respectively, mRNA and protein expression levels of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR), cholesterol 7α-hydroxylase 1 (CYP7A1), LDL-R, and sterol regulatory element binding protein-2 (SREBP-2) were detected using real time RT-PCR and Western blotting.
RESULTSCompared with the blank control group, serum levels of TC (1.84 ± 0.19 mmol/L, 2.23 ± 0.43 mmol/L) and LDL-C (0.99 ± 0.24 mmol/L, 1.13 ± 0.56 mmol/L) were higher in the hyperlipidemia group and the PDS hyperlipidemia group, serum levels of HDL-C (0.41 ± 0.66 mmol/L, 0.41 ± 0.11 mmol/L) and AMY activities (351 ± 45 mmol/L, 153 ± 30 mmol/L) were lower, and urinary D-xylose excretion rates were lower (26.9 ± 2.1 ng/mL, 15.0 ± 1.7 ng/mL) (all P < 0.05). Lipid deposition occurred in liver cells. Much fat vacuoles occurred in the cytoplasm. Expression levels of HMGCR, CYP7A1, LDL-R, and SREBP-2 mRNA and proteins in liver significantly decreased (P < 0.01). Compared with the hyperlipidemia group, serum levels of TC and LDL-C significantly increased (P < 0. 05), AMY activities and urinary D-xylose excre- tion rates significantly decreased in the PDS hyperlipidemia group (P < 0.01). A large amount of lipid deposition occurred in liver. The atrophy of liver cells was obviously seen. Expression levels of CYP7A1, LDL-R, and SREBP-2 mRNA and proteins in liver were significantly lower (P < 0.01, P < 0.05). Serum levels of TC and LDL-C significantly decreased (P < 0.05), AMY activities and urinary D-xylose excretion rates significantly increased in the hyperlipidemia treatment group (P < 0.01). Expression levels of CYP7A1, LDL-R, and SREBP-2 mRNA and proteins in liver were significantly increased (P < 0.01, P < 0.05). Compared with the PDS hyperlipidemia group, serum level of TC significantly decreased (P < 0.05), HDL-C levels, AMY activities and urinary D-xylose excretion rates significantly increased in the PDS hyperlipidemia treatment group (P < 0.01),expression levels of CYP7A1, LDL-R, and SREBP-2 mRNA and proteins in liver were significantly increased (P < 0.01). Similar changes occurred in the two treatment groups.
CONCLUSIONSPi deficiency exacerbates abnormal serum TC level and the lipid deposition in liver. These might be related to regulating expression levels of LDL-R, HMGCR, and CYP7A1 genes in the SREBP-2 signal pathway. HQR could regulate this pathway to intervene abnormal metabolism of TC.
Animals ; Cholesterol, HDL ; Cholesterol, LDL ; Drugs, Chinese Herbal ; therapeutic use ; Hyperlipidemias ; drug therapy ; Liver ; Male ; Medicine, Chinese Traditional ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Sterol Regulatory Element Binding Protein 2 ; metabolism ; Triglycerides
2.Effect and mechanism of total flavonoids of bugloss on rats with myocardial ischemia and reperfusion injury.
Xiao-Na XU ; Zi-Ran NIU ; Shou-Bao WANG ; Yu-Cai CHEN ; Li GAO ; Lian-Hu FANG ; Guan-Hua DU
Acta Pharmaceutica Sinica 2014;49(6):875-881
This study is to investigate the effect of total flavonoids of Uygur medicine bugloss (BTF) on rats with myocardial ischemia/reperfusion injury, and to explore the mechanisms by which it acts. Left anterior descending (LAD) coronary artery in rats was occluded for 30 min followed by 4 h reperfusion. Meanwhile, BTF dissolved in saline was administered intraperitoneally at dosage of 10, 30 and 50 mg x kg(-1). Electrocardiograph, infarction index, serum myocardial enzymes and heart function were determined to evaluate the effect of BTF. Some other observations were carried out to explore whether inhibiting inflammation and apoptosis is involved in the mechanisms underlying BTF. Our results showed that in ischemia/reperfusion injured rats BTF could dose-dependently reduce myocardial infarction index and myocardial enzyme leakage, and enhance heart function, indicating that it possesses significant cardio protection. ELISA analysis showed that BTF could decrease the content of myocardial inflammatory cytokines such as IL-1beta, IL-6 and TNF-alpha. Western-blotting confirmed that BTF could increase the expression of anti-apoptotic protein Bcl-2 and reduce the expression of proapoptosis protein Bax. Further more, the phosphorylation level of PI3K and Akt was upregulated by BTF treatment. BTF can protect rat against myocardial ischemia/reperfusion injury. Anti-inflammation and inhibition of apoptosis through upregulating PI3K/Akt signal pathway may contribute to the protective effect of BTF.
Animals
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Apoptosis
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Apoptosis Regulatory Proteins
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Boraginaceae
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chemistry
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Flavonoids
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pharmacology
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Heart
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Interleukin-6
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Myocardial Infarction
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Myocardial Reperfusion Injury
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drug therapy
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Myocardium
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Phosphatidylinositol 3-Kinases
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Phosphorylation
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Protective Agents
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Proto-Oncogene Proteins c-akt
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Rats
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Signal Transduction
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Tumor Necrosis Factor-alpha
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bcl-2-Associated X Protein
3.Construction of eukaryotic expression vector of E4F1 and interactions between E4F1 and p53
Panfeng LIAN ; Long CHENG ; Xin GUAN ; Dayang ZOU ; Ling MEI ; Yuan SHEN ; Wei REN ; Juhui ZHANG ; Qinong YE ; Enqun WANG
Military Medical Sciences 2014;(1):53-56
Objective To construct eukaryotic expression vector of wild type E 4F1 and the mutant deleting amino acid region 32-81, and to detect the interaction between wild type or mutant E 4F1 and p53 and to study the effect of E4F1 on the expression level of p21.Methods Wild type and mutant sequences of E 4F1 were amplified from the mammary library using standard PCR and recombinant PCR .The sequences were cloned into pXJ 40-MYC vector to generate the MYC-E4F1 and MYC-E4F1(Δ32-81) recombinant plasmids that were transfected into 293T cells and identified by Western blotting . FLAG-p53 and MYC-E4F1 or MYC-E4F1(Δ32-81) were co-transfected into 293T cells and immunoprecipitation assay was performed to detect the interaction of wild type or mutant E 4F1 with p53.Wild type and mutant E4F1 expressing vec-tors were co-transfected into osteosarcoma U2OS cells and the expression of p21was detected.Results Recombinant plas-mids of MYC-E4F1 and MYC-E4F1(Δ32-81) were successfully constructed.Both wild type and mutant E4F1 interacted with p53.Deletion of amino acid region 32-81 of E4F1 increased the interaction .The expression level of p21 was in-creased by wild-type E4F1, but not by mutant E4F1.Conclusion The eukaryotic expression vector of wild type E4F1 and its deletion mutant is successfully constructed .Both of them interact with p53.Deletion of amino acid region 32-81 of E4F1 increases the interaction .This study contributes to further studies on the regulation and mechanism of E 4F1 on p53.
4.Puerarin reduces oxidative damage to vascular endothelial cells by improving mitochondrial respiratory function
Shu-chan SUN ; Di-fei GONG ; Tian-yi YUAN ; Shou-bao WANG ; Lian-hua FANG ; Guan-hua DU
Acta Pharmaceutica Sinica 2022;57(5):1352-1360
This study investigated the effect of puerarin on human umbilical vein endothelial cells (HUVEC) injured with hydrogen peroxide (H2O2). HUVEC were divided into three groups: a control group, a model group (H2O2 400 μmol·L-1) and a puerarin-treated group (3, 10, 30 and 100 μmol·L-1). HUVEC were cultured with varied concentration of puerarin for 2 h and treated with H2O2 for another 24 h. Cell proliferation was detected by a CCK-8 assay. The mitochondrial membrane potential was measured by a JC-1 fluorescent probe. A transwell chamber assay was adopted to observe cell migration ability. Mitochondrial respiratory function was measured in a two-chamber titration injection respirometer (Oxygraph-2k). The expression of interleukin-1
5.Protective effect of sodium pyruvate on ischemia/reperfusion injury of rats subjected to hemorrhagic shock.
Li-Dong GUAN ; Zi-Ling WANG ; Lian ZHAO ; Bo WANG ; Guang-Yi WANG ; Guang-Zhi WEI ; Hong ZHOU
Chinese Journal of Applied Physiology 2007;23(3):264-268
AIMTo study the protective effect of sodium pyruvate on ischemia/reperfusion injury following hemorrhagic shock.
METHODSRat models of hemorrhagic shock were built up. When the shed blood was infused, the rats were also randomly provided by one of normal saline, glutathione and sodium pyruvate. Rats were killed 3 hours after the reperfusion, the activity of plasma lactate dehydrogenase (LDH) and glutamic-oxaloacetic transaminase (GOT), the level of tissue malondialdehyde (MDA) and the activity of tissue myeloperoxidase (MPO) were detected. Biopsy specimens were obtained to investigate morphological changes of the myocardial, hepatic, lung and renal tissue.
RESULTSThe activity of plasma LDH and GOT, the level of MDA of hepatic, lung and renal tissue and the activity of MPO of myocardial, lung and renal tissue decreased remarkably in group given sodium pyruvate compared with group given normal saline, and the effect of group given sodium pyruvate was more remarkable than group given glutathione.
CONCLUSIONThese data support the view that sodium pyruvate shows protective effect on ischemia/reperfusion injury following hemorrhagic shock. It is possibly relevant to scavenging of oxygen free radicals, reduction of neutrophil, and anti-inflammatory response.
Animals ; Aspartate Aminotransferases ; blood ; Disease Models, Animal ; Kidney ; metabolism ; pathology ; L-Lactate Dehydrogenase ; blood ; Liver ; metabolism ; pathology ; Lung ; metabolism ; pathology ; Male ; Malondialdehyde ; analysis ; Peroxidase ; analysis ; Protective Agents ; pharmacology ; Pyruvic Acid ; pharmacology ; Rats ; Rats, Wistar ; Reperfusion Injury ; blood ; pathology ; prevention & control ; Shock, Hemorrhagic ; blood ; pathology
6.Reflectance confocal microscopy features of several common diseases manifesting as papules in children
Lixin CHEN ; Haihui SU ; Ying WANG ; Ji WANG ; Zekun GONG ; Zhiwei GUAN ; Jia LIAN ; Xiaoyan FENG ; Yangyang LIN ; Xibo GAO ; Xinxin LIU ; Tiantian BI ; Qinfeng LI
Chinese Journal of Dermatology 2016;49(11):817-820
Objective To evaluate the clinical application value of reflectance confocal microscopy(RCM) in the diagnosis of several common diseases manifesting as papules in children, including lichen nitidus, verruca planae, lichen striatus, milium, molluscum contagiosum and lichen pilaris. Methods A total of 579 children clinically characterized by papules were recruited into this study. RCM was used to observe lesions and perilesional normal skin. The RCM features of 6 diseases manifesting as papules were analyzed and compared. Results Based on RCM images, 236 patients were diagnosed with lichen nitidus, 70 with verruca planae, 123 with lichen striatus, 40 with milium, 53 with molluscum contagiosum and 57 with lichen pilaris. All the 6 diseases had typical RCM features. Concretely speaking, RCM images of lichen nitidus lesions showed infiltration of dense inflammatory cells and melanophages in enlarged dermal papillae. In RCM images of verruca planae lesions, cells in the granular and spinous layers were arranged in concentric circles, giving a rose cluster?like appearance. RCM images of lichen striatus lesions revealed focal swelling of stratum spinosum, absent or local liquifaction degeneration of basal cells, and clustering of a moderate number of inflammatory cells in the superficial dermis. In RCM images of milium lesions, well?circumscribed round or oval structures containing highly but nonuniformly refractive materials could be seen in the dermis. RCM images of molluscum contagiosum lesions showed intact cystoid structures containing highly refractive molluscum bodies. Lowly to moderately refractive cutin ? like materials were observed along with the dilation of hair follicle infundibula in RCM images of lichen pilaris lesions. In RCM images, the 6 diseases were distinguished mainly based on structural features(patterns and refractivity)of skin lesions shown by continuous vertical scanning. Conclusion RCM is of great value to the diagnosis of diseases manifesting as papules in children.
7.Puerarin protects against myocardial ischemia/reperfusion injury by suppressing NLRP3 inflammasome activation via TLR4/Myd88/NF-κ B pathway in rats
Dan-shu WANG ; Liu-yan YAN ; Shu-chan SUN ; Yu JIANG ; Su-yue YIN ; Shou-bao WANG ; Lian-hua FANG ; Guan-hua DU
Acta Pharmaceutica Sinica 2021;56(5):1343-1351
This study was to investigate the protective effects of puerarin on myocardial ischemia/reperfusion (MI/R) injury and the underlying mechanism. The MI/R-model was established by ligating the left anterior descending artery (LAD) for 60 min followed by 24 h reperfusion, puerarin (10, 30, and 100 mg·kg-1) was orally administered 20 min before reperfusion. Cardiac function, myocardial infarct index, cardiac damage markers, inflammatory cytokines, and apoptosis index were measured to evaluate the protective effects of puerarin on MI/R injury. The activation of Nod-like receptor protein 3 (NLRP3) inflammasome and Toll like receptor 4 (TLR4)/myeloid differentiation factor 88 (Myd88)/nuclear factor kappa B (NF-
8.Vasorelaxant effect and mechanisms of compound reserpine and triamterene tablets on the isolated thoracic aorta rings
Di-fei GONG ; Ran-ran WANG ; Tian-yi YUAN ; Shou-bao WANG ; Jun-ke SONG ; Lian-hua FANG ; Guan-hua DU
Acta Pharmaceutica Sinica 2022;57(11):3339-3344
This study aimed to evaluate the vasorelaxant effect and mechanisms of compound reserpine and triamterene tablets (CRTTs) and its component triamterene on isolated rat thoracic aorta rings. Isolated rat thoracic aorta rings pre-contracted by high potassium or norepinephrine (NE) were used to evaluate the vasodilatory effect of CRTTs and its component triamterene. The mechanisms concerning endothelium, potassium channels and calcium channels were studied through the interventions of several tool drugs. Animal welfare and experimental procedures followed the requirements of the Laboratory Animal Management and Animal Welfare Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College. The results showed that both CRTTs and triamterene had potent relaxant effect on KCl and NE pre-contracted vessels. Triamterene showed partial endothelium dependency, and
9.The efficacy and safety of intravenous bisphosphonates in the treatment of primary hyperparathyroidism complicated by hyperculcemia crisis
Guiyan HAN ; Ou WANG ; Xiaoping XING ; Xunwu MENG ; Xiaolan LIAN ; Heng GUAN ; Wei YE ; Weibo XIA ; Mei LI ; Yan JIANG ; Yingying HU ; Huaicheng LIU ; Quancai CUI
Chinese Journal of Internal Medicine 2009;48(9):729-733
creatinine concentration. Conclusion Bisphosphonates can decrease serum total calcium levels in hypercalcemia crisis caused by PHPT effectivelywith mild adverse events.
10.Clinical characteristics and microbiological tests analysis of 18 patients with Talaromyces marneffei infection
ZOU Sheng-hua ; FANG Mei-lan ; LIN Zhen-li ; CHEN Xin-chao ; HUANG Ming-xiang ; GUAN Hong-lian ; WANG Xin-hang
China Tropical Medicine 2023;23(2):139-
Abstract:Objective To investigate the clinical characteristics and early diagnostic methods of patients with Talaromyces marneffei infection, so as to reduce the mortality of patients. Methods The clinical characteristics and microbiological analysis data including fungal culture, smear examination and mass spectrometry were collected from 18 patients with Talaromyces marneffei infection in the Department of Respiratory Medicine, Department of Tuberculosis, and Department of Critical Respiratory Medicine in Fuzhou Pulmonary Hospital from January 2017 to December 2021, and descriptive analysis was conducted. Results All the 18 patients were confirmed to be infected with Talaromyces marneffei by conventional culture and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS). The main infection sites of 18 patients with Talaromyces marneffei infection were lungs and lymph nodes, and the patients were accompanied by clinical manifestations such as cough, sputum and fever. The imaging features such as patchy shadows, mediastinal lymph node shadows and nodular shadows were common. Microbiological testing showed a statistically significant difference between smear and culture with a higher positive culture rate (χ2=13.74, P<0.05). The positive rate of blood culture in microbiological test was 60.0% (9/15), the positive rate of bronchial lavage fluid culture was 26.7% (4/15), the positive rate of sputum culture was 5.6% (1/18), one case each of pus, bone marrow, pleural fluid and cerebrospinal fluid was positive for culture and the other cases were negative, one case of sputum and one case of pus were positive for smear and the rest were negative. Colony characteristics showed that the colony morphology was mycelial phase at 25 ℃, producing red pigment, and the branching pattern of the penicillus was seen microscopically as monoverticillate or biverticillate; At 35 ℃, the yeast phase appeared at the initial stage, and then the mycelium phase changed after 5-6 days; the yeast phase was observed at 37 ℃, and yeast-like cells were seen under the microscope. All 18 patients with Talaromyces marneffei infection got better after using antifungal drugs. Compared with non-HIV patients with Talaromyces marneffei infection, leukopenia and anemia were common in HIV patients with Talaromyces marneffei infection, and the differences were statistically significant (P<0.05). Conclusions The infection of Talaromyces marneffei can be divided into localized type and disseminated type, which usually invade the lungs, skin, lymph nodes and other places. The main manifestations of patients are fever, cough, phlegm and other atypical symptoms. At present, the diagnosis of Talaromyces marneffei infection is mostly based on the fungal culture test, and the application of MALDI-TOF MS method can effectively shorten the diagnosis time of Talaromycosis marneffei. Clinical characteristics combined with microbiological analysis provide an objective basis for early diagnosis of patients with Talaromyces marneffei infection, and timely use of antifungal therapy can improve the prognosis of patients.