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1.
Yonsei Medical Journal ; : 262-273, 2021.
Article in English | WPRIM | ID: wpr-875610

ABSTRACT

Purpose@#This study aimed to explore the role of the long non-coding RNA (lncRNA) RNA component of mitochondrial RNAase P (RMRP) in sepsis-induced acute kidney injury (AKI). @*Materials and Methods@#Venous blood was collected from septic patients and healthy people. C57BL/6 mice who underwent cecal ligation and puncture (CLP) were used as in vivo models of septic AKI. Lipopolysaccharide (LPS)-induced HK-2 cells were employed as in vitro models of AKI. Flow cytometry analysis was conducted to detect cell apoptosis. Enzyme-linked immunosorbent assay and Western blot assays were used to detect levels of pro-inflammatory cytokines. @*Results@#RMRP was upregulated in sera from patients with AKI and in LPS-induced cells. Knockdown of RMRP inhibited cell apoptosis and reduced production of inflammatory factors in LPS-induced cells, as well as alleviated AKI in CLP mice. RMRP facilitated inflammation by activating NACHT, LRR, and PYD domains-containing protein 3 (NLRP3) inflammasome. We found that microRNA 206 (miR-206) binds with and is negatively regulated by RMRP: miR-206 directly targets the 3’ untranslated region of DEAD-box helicase 5 (DDX5) and negatively regulates DDX5 expression. By binding with miR-206, RMRP upregulated DDX5 expression. Rescue assays revealed that overexpression of DDX5 counteracted the effect of RMRP inhibition on cell apoptosis and inflammatory response in LPS-induced cells. @*Conclusion@#The lncRNA RMRP contributes to sepsis-induced AKI through upregulation of DDX5 in a miR-206 dependent manner and through activation of NLRP3 inflammasome. This novel discovery may provide a potential strategy for treating AKI.

2.
Article in Chinese | WPRIM | ID: wpr-879544

ABSTRACT

OBJECTIVE@#To explore the genetic basis for Chinese pedigree affected with tuberous sclerosis complex (TSC).@*METHODS@#The proband and his family members were subjected to Sanger sequencing for variants of the TSC1 and TSC2 genes.@*RESULTS@#The proband was found to harbor a c.2837+1dupG splicing variant at a donor site of the TSC2 gene. The same variant was not found among his family members and the fetus during his mother's subsequent pregnancy.@*CONCLUSION@#The c.2837+1dupG splicing variant of the TSC2 gene has probably predisposed to the TSC in this pedigree. Above finding has enriched the spectrum of pathogenic variants associated with this disease.


Subject(s)
Female , Genetic Testing , Humans , Male , Mutation , Pedigree , Pregnancy , Prenatal Diagnosis , Tuberous Sclerosis/genetics , Tuberous Sclerosis Complex 2 Protein/genetics
3.
Article in Chinese | WPRIM | ID: wpr-883966

ABSTRACT

Objective:To explore the correlation between hypersomnia and anhedonia in patients with major depressive disorder.Methods:From November 2018 to May 2019, patients hospitalized with major depressive disorder who met the ICD-10 diagnostic criteria were selected.According to the Epworth Sleepiness Scale (ESS), 46 patients were divided into daytime sleepiness group with ESS ≥ 7, and 171 patients were divided into non-sleepiness group with ESS < 7.The Chinese Revised Social Anhedonia Scale (RSAS) and the Chinese Revised Physical Anhedonia Scale (RPAS) were used to evaluate the patients' anhedonia symptoms.Two-way ANOVA and Pearson correlation analysis were used for data processing.Results:(1)There was no interaction between the hypersomnia and gender on the score of physical anhedonia ( F=0.274, P=0.601). The main effect analysis showed that there was significant difference in the influence of gender on physical anhedonia ( F=10.948, P<0.05). (2)There was interaction between the hypersomnia and age on the score of physical anhedonia ( F=4.396, P=0.013). Further simple effect analysis showed that the score of physical anhedonia in 40-49 age(21.54±12.37) was lower than that in 50-64 age(34.13±12.53) in daytime sleepiness group( P<0.05). (3) There was interaction between hypersomnia and sitting and lying on the score of social anhedonia ( F=4.247, P=0.041). Further simple effect analysis showed that the score of social anhedonia in patients with sitting and lying time less than 2 hours (13.71±5.18) was lower than that in patients with sitting and lying time more than 2 hours (19.75±6.39) in daytime sleepiness group( P<0.05). (4)Pearson correlation analysis showed that the total sleepiness score of depression patients was positively correlated with the social anhedonia score ( r=0.206, P<0.01). After adjusting for gender, age and sitting and lying time, the total sleepiness score was still positively correlated with the social anhedonia score( r=0.225, P<0.01). Conclusion:Hypersomnia may be associated with anhedonia in patients with major depressive disorder.

4.
Article in Chinese | WPRIM | ID: wpr-883640

ABSTRACT

In order to explore the application of the teaching mode based on WeChat official platform in the teaching of experimental pathology for international students, the students are required to browse the teaching objectives and the general pictures and microscopic pictures of experimental specimens on the public platform before the experimental class, and get in touch with the preliminary diagnosis of pathology at an early stage, contact with the preliminary diagnosis of pathology early, and consolidate and expand their knowledge after class by using the platform. The research shows that the mixed teaching mode based on the official platform can improve the diagnostic thinking of foreign students in pathology, deepen their understanding of pathology, increase the interaction between teachers and students and the supervision of teachers on students, and enhance their interest in learning.

5.
Article in Chinese | WPRIM | ID: wpr-883517

ABSTRACT

Three hundred and twenty endophytic actinobacterial strains were isolated from psammophytes collected from Taklamakan Desert and identified.Among them,three strains already had been identified as new species of two genera and sixteen isolates showed relatively low 16S rRNA similarities<98.6%to validly described species.Seventy-five of the isolates were selected as representative strains to screen antibacterial activity and mechanism.Forty-seven strains showed antagonistic activity against at least one of the indicator bacteria.Two Streptomyces strains produced bioactive compounds inducing DNA damage,and two Streptomyces strains produced bioactive compounds with inhibitory activity on protein biosynthesis.Notably,the strain Streptomyces sp.8P21H-1 that demonstrated both strong antibacterial activity and inhibitory activity on protein biosynthesis was prioritized for exploring new antibiotics.Under the strategy of integrating genetics-based discovery program and MS/MS-based molecular networking,two new streptogramin-type antibiotics,i.e.,acetyl-griseoviridin and desulphurizing gri-seoviridin,along with known griseoviridin,were isolated from the culture broth of strain 8P21H-1.Their chemical structures were determined by HR-MS,and 1D and 2D NMR.Desulphurizing griseoviridin and griseoviridin exhibited antibacterial activities by inhibiting translation.

7.
Article in Chinese | WPRIM | ID: wpr-879892

ABSTRACT

OBJECTIVE@#To study the association between maternal reduced folate carrier (@*METHODS@#A hospital-based case-control study was conducted. The mothers of 683 infants with CHD who attended the Department of Cardiothoracic Surgery, Hunan Children's Hospital, from November 2017 to March 2020 were enrolled as the case group. The mothers of 740 healthy infants without any deformity who attended the hospital during the same period of time were enrolled as the control group. A questionnaire survey was performed to collect the exposure data of subjects. Venous blood samples of 5 mL were collected from the mothers for genetic polymorphism detection. A multivariate logistic regression analysis was used to evaluate the association of @*RESULTS@#After control for confounding factors, the multivariate logistic regression analysis showed that maternal @*CONCLUSIONS@#Maternal


Subject(s)
Case-Control Studies , Child , Female , Genetic Predisposition to Disease , Genotype , Heart Defects, Congenital/genetics , Humans , Infant , Polymorphism, Single Nucleotide , Reduced Folate Carrier Protein/genetics , Risk Factors
8.
Article in Chinese | WPRIM | ID: wpr-879198

ABSTRACT

Drug combination is a common clinical phenomenon. However, the scientific implementation of drug combination is li-mited by the weak rational evaluation that reflects its clinical characteristics. In order to break through the limitations of existing evaluation tools, examining drug-to-drug and drug-to-target action characteristics is proposed from the physical, chemical and biological perspectives, combining clinical multicenter case resources, domestic and international drug interaction public facilities with the aim of discovering the common rules of drug combination. Machine learning technology is employed to build a system for evaluating and predicting the rationality of clinical drug combinations based on "drug characteristics-repository information-artificial intelligence" strategy, which will be debugged and validated in multi-center clinical practice, with a view to providing new ideas and technical references for the safety and efficacy of clinical drug use.


Subject(s)
Artificial Intelligence , Drug Combinations , Machine Learning , Technology
9.
Article in Chinese | WPRIM | ID: wpr-879188

ABSTRACT

The metabolites of salvianolic acid A and salvianolic acid B in rats were analyzed and compared by ultra-high-perfor-mance liquid chromatography with linear ion trap-orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS). After the rats were administrated by gavage, plasma at different time points and urine within 24 hours were collected to be treated by solid phase extraction(SPE), then they were gradient eluted by Acquity UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) and 0.1% formic acid solution(A)-acetonitrile(B) mobile phase system, and finally all biological samples of rats were analyzed under negative ion scanning mode. By obtaining the accurate relative molecular mass and multi-level mass spectrometry information of metabolites, combined with the characteristic cleavage law of the reference standard and literature reports, a total of 30 metabolites, including salvianolic acid A and B, were identified. Among them, there were 24 metabolites derived from salvianolic acid A, with the main metabolic pathways including ester bond cleavage, dehydroxylation, decarboxylation, hydrogenation, methylation, hydroxylation, sulfonation, glucuronidation, and their multiple reactions. There were 15 metabolites of salvianolic acid B, and the main biotransformation pathways were five-membered ring cracking, ester bond cleavage, decarboxylation, dehydroxylation, hydrogenation, methylation, sulfonation, glucuronidation, and their compound reactions. In this study, the cross-metabolic profile of salvianolic acid A and B was elucidated completely, which would provide reference for further studies on the basis of pharmacodynamic substances and the exploration of pharmacological mechanism.


Subject(s)
Animals , Benzofurans , Caffeic Acids , Chromatography, High Pressure Liquid , Lactates , Mass Spectrometry , Rats , Technology
10.
Article in Chinese | WPRIM | ID: wpr-879186

ABSTRACT

Non-alcoholic steatohepatitis(NASH) was induced by high-sugar and high-fat diet in mice to investigate the intervention effect of total saponins from Panax japonicus(TSPJ) and explore its possible mechanism. Mice were fed with high-sugar and high-fat diet to establish NASH model, and intervened with different doses of TSPJ(15, 45 mg·kg~(-1)). The animals were fed for 26 weeks. The histomorphology and pathological changes of liver tissues were observed by HE staining. The transcriptional expression levels of miR-199 a-5 p, autophagy related gene 5(ATG5) and inflammatory cytokines interleukin-6(IL-6), interleukin-1β(IL-1β) and tumor necrosis factor α(TNF-α) in mouse liver were measured by quantitative Real-time polymerase chain reaction(qRT-PCR). Western blot was used to detect the expression of autophagy-related proteins ATG5, P62/SQSTM1(P62), and microtubule-associated protein light chain 3(LC3)-I/Ⅱ proteins in mouse liver. The expression of P62 protein was detected by immunofluorescence staining. In order to verify the targeting regulation relationship between miR-199 a-5 p and ATG5, miR mimic/inhibitor NC and miR-199 a-5 p mimic/inhibitor were transfected into Hepa 1-6 cells, and the expression of ATG5 mRNA and protein was detected. pMIR-reportor ATG5-3'UTR luciferase reporter gene plasmid was constructed and co-transfected with miR mimic/inhibitor NC and miR-199 a-5 p mimic/inhibitor into Hepa 1-6 cells to detect luciferase activity. In vivo, HE staining in the model group showed typical fatty degeneration and inflammatory infiltration, with increased expression of miR-199 a-5 p and decreased expression of ATG5 mRNA and protein. The expression of autophagy-associated protein P62 increased significantly, the ratio of LC3Ⅱ/Ⅰ decreased, and the transcriptional expression of inflammatory factors increased significantly. After the intervention by TSPJ, the pathological performance of liver tissue was significantly improved, the expression of miR-199 a-5 p decreased and the expression of ATG5 mRNA and protein increased, the expression of autophagy-associated protein P62 decreased significantly, the ratio of LC3Ⅱ/Ⅰ increased, and the transcriptional expression of inflammatory cytokines IL-6, IL-1β and TNF-α decreased significantly. In vitro, it was found that the expression of ATG5 mRNA and protein and luciferase activity decreased significantly in miR-199 a-5 p overexpression cells, while after inhibition of miR-199 a-5 p expression, the expression level of ATG5 mRNA and protein and luciferase activity increased. The results showed that TSPJ can improve NASH in mice fed with high-sugar and high-fat diet, and its mechanism may be related to the regulation of miR-199 a-5 p/ATG5 signal pathway, the regulation of autophagy activity and the improvement of inflammatory response of NASH.


Subject(s)
Animals , Autophagy , Autophagy-Related Protein 5 , Mice , MicroRNAs/genetics , Non-alcoholic Fatty Liver Disease/genetics , Panax , Saponins/pharmacology
11.
Article in Chinese | WPRIM | ID: wpr-879142

ABSTRACT

To probe the potential hepatotoxic components of Epimedii Folium and investigate its mechanism based on network toxicology and cell experimental validation. According to the previous results of component measurement and cytotoxicity evaluation, 11 active compounds related to hepatotoxicity in Epimedii Folium were chosen as research object in this study. Through SwissTargetPrediction database and GeneCards database, the potentially hepatotoxic targets of Epimedii Folium were obtained. Subsequently, the protein-target interaction network and active compounds-hepatotoxic targets network were established to analyze the core targets and screen the key hepatotoxic compounds in Epimedii Folium. Meanwhile, the signaling pathways and molecular mechanisms were inferred with GO functional enrichment analysis and KEGG pathway enrichment analysis on the core targets. At last, the effect of icaritin as the chief hepatotoxic compound on the indexes related to hepatotoxicity in HL-7702 cells and HepG2 cells was investigated to validate the hepatotoxicity mechanism of Epimedii Folium. Through the network toxicology analysis, 190 action targets and 991 hepatotoxic targets were collected, then 64 potentially hepatotoxic targets of Epimedii Folium including AKT1, EGFR, MAPK3, TNF and so on were obtained, and icaritin was screened as the key hepatotoxic compound. GO functional enrichment analysis indicated 160 biological process terms such as protein phosphorylation and negative regulation of apoptotic process, 41 molecular function terms such as protein binding and ATP binding, and 32 cellular component terms such as cytosol and cell surface. KEGG pathway enrichment analysis inferred 75 signaling pathways involving PI3 K-Akt and HIF-1. After comprehensive analysis, it was inferred that the hepatotoxicity mechanism of Epimedii Folium was related with regulating oxidative stress and apoptosis. The results of cell biology experiments showed that icaritin could significantly increase the level of aspartate aminotransferase and lactate dehydrogenase, reduce the level of glutathione, improve the quality of reactive oxygen species and reduce mitochondrial membrane potential, indicating that it could cause hepatotoxicity by destroying cell membrane structure, inhibiting antioxidant enzyme activity, activating oxidative stress and inducing apoptosis. These results proved the reliability of results of network pharmacology. This study preliminarily clarified the material base and the mechanism of potential hepatotoxicity of Epimedii Folium, which provided important information for further research and safe application.


Subject(s)
Drugs, Chinese Herbal/toxicity , Plant Leaves , Protein Interaction Maps , Reproducibility of Results
12.
Article in Chinese | WPRIM | ID: wpr-879075

ABSTRACT

Dendrobii officinalis, with a definite effect of nourishing Yin and clearing heat, has been a folk habit for drinking after being mixed with water. Because its superfine powder has the advantages of high dissolution and convenient drinking, we observed the effect of D. officinalis superfine powder on metabolic hypertension model rats and its possible mechanism in this experiment, which can be used as a reference for its clinical application for hypertension. The overeating greasy-induced metabolic hypertension model was established with high-fat, high-sugar and high-purine diet. These rats were orally administered with 400 mg·kg~(-1) and 200 mg·kg~(-1) of D. officinalis superfine powder for 20 consecutive weeks. During this period, blood pressure, blood lipid, blood glucose, insulin and other related indexes of glucose and lipid metabolism were monitored; the levels of lipopolysaccharide(LPS), C-reactive protein(CRP), interleukin 6(IL-6) and other inflammatory mediators were measured; the levels of nitric oxide(NO) and endothelin-1(ET-1) were detected, and the histomorphological and ultrastructural changes of aorta were observed. In addition, the expression of LPS/TLR4 pathway-related molecules in aorta was determined. The results showed that long-term administration of D. officinalis superfine powder significantly reduced the levels of systolic blood pressure(SBP), diastolic blood pressure(DBP) and mean arterial pressure(MBP) in metabolic hypertension model rats, decreased the levels of total cholesterol(TC), triglyceride(TG), low density lipoprotein cholesterol(LDL-c), glucose(Glu), and insulin(INS) levels in blood, increased the contents of high density lipoprotein cholesterol(HDL-c),decreased the LPS, CRP, IL-6 and ET-1 levels in blood and increased NO content. Furthermore, it improved the abnormality of aortic histomorphology and endothelial ultrastructure, and inhibited the protein expression of TLR4, myeloid differentiation factor(MyD88), IL-6, interleukin-1 β(IL-1β), and tumor necrosis factor-α(TNF-α) as well as mRNA expression of TNF-α and IL-1β in aorta. In conclusion, D. officinalis superfine powder may improve the abnormal function and structure of blood vessels by inhibiting the activation of LPS/TLR4 pathway, thus playing a role against metabolic hypertension.


Subject(s)
Animals , Dendrobium/chemistry , Drugs, Chinese Herbal/pharmacology , Hyperphagia , Hypertension/drug therapy , Interleukin-6 , Powders , Rats , Tumor Necrosis Factor-alpha
13.
Article in Chinese | WPRIM | ID: wpr-878912

ABSTRACT

Chinese medicinal material is the foundation of traditional Chinese medicine(TCM) industry. Its quality is not only closely related to the health of residents but also the key to the development of the TCM industry. Pesticide residues, heavy metals and mycotoxins are the major pollutants of Chinese medicinal materials. In recent years, quite a number of rapid detection methods for pollutants have been constructed. Among them, surface-enhanced Raman scattering(SERS), which has been widely used in food chemistry, environmental analysis, and other fields because of its speediness and non-destructiveness, shows its great potential in the pollutant detection in Chinese medicinal material. This paper firstly reviews the application of SERS for the detection of common pollutants in Chinese medicinal material. We then discussed the characteristics and advantages of SERS technique for pesticide detection, including the principle, SERS substrate design, specific recognition, etc. Finally, simultaneous detection of multiple pesticide residues in Chinese medicinal material was explored.


Subject(s)
China , Environmental Pollutants , Medicine, Chinese Traditional , Pesticide Residues/analysis , Spectrum Analysis, Raman
14.
Article in English | WPRIM | ID: wpr-877728

ABSTRACT

INTRODUCTION@#Associations of variations in PLA2R1 and HLA-DQA1 genes with susceptibility to idiopathic membranous nephropathy (IMN) have been well documented. Association with spontaneous remission, however, is poorly defined in the Chinese Han population.@*METHODS@#A Chinese cohort of 117 IMN patients and 138 healthy controls were recruited between July 2009 and November 2019. Case-control studies for single-nucleotide polymorphisms (SNPs) within HLA-DQA1 (rs2187668) and PLA2R1 (rs35771982, rs4664308, rs3749117, rs3749119) genes were performed. The contributions of these polymorphisms to predict susceptibility, titre of autoantibodies against the M-type phospholipase A2 receptor (anti-PLA2R1), glomerular PLA2R1 expression, and spontaneous remission were analysed.@*RESULTS@#We found that variations in PLA2R1 (SNPs rs35771982, rs4664308, rs3749117) were strongly associated with IMN susceptibility, while SNP (rs2187668) within HLA-DQA1 did not increase the risk of IMN. All SNPs in PLA2R1 and HLA-DQA1 were not statistically associated with anti-PLA2R1 titre, glomerular PLA2R1 expression and spontaneous remission after Bonferroni correction (@*CONCLUSION@#This study confirms that variations in PLA2R1 (SNPs rs35771982, rs4664308, rs3749117) are risk factors for IMN. We found excellent association of serum albumin level, anti-PLA2R1 titre and glomerular PLA2R1 positivity with non-spontaneous remission in IMN.

15.
Acta Pharmaceutica Sinica ; (12): 723-733, 2021.
Article in Chinese | WPRIM | ID: wpr-876513

ABSTRACT

Indoleamine 2,3-dioxygenase 1 (IDO1) is the rate-limiting enzyme in the degradation of tryptophan to kynurenine. IDO1 is highly expressed in some tumor tissues. IDO1 can deplete tryptophan in tumor microenvironment, inhibit T cell function, and mediate the immune escape of tumor cells. Thus, IDO1 is considered a potential target of tumor immunotherapy. Currently, there are several IDO1 inhibitors in clinical research studies. The mechanism of IDO1-mediated tumor immune escape and the structure of IDO1 inhibitors are summarized in this review.

16.
Article in Chinese | WPRIM | ID: wpr-873233

ABSTRACT

Objective:To prepare Periplaneta americana thermosensitive hydrogel and investigate its effect on wound healing in diabetic rats. Method:Taking N-isopropylacrylamide (NIPAM) and acrylic acid (AAc) as monomers, thermosensitive poly(NIPAM-co-AAc) [P(NIPAM-co-AAc)] polymeric material was prepared by free radical polymerization, then thermoresponsive copolymer P(NIPAM-co-AAc)-g-HA was synthesized by conjugating P(NIPAM-co-AAc) to hyaluronic acid (HA). The structure and lower critical solution temperature (LCST) of the graft copolymer were characterized by proton nuclear magnetic resonance spectroscopy (1H-NMR) and ultraviolet spectrophotometry (UV). P. americana thermosensitive hydrogel was prepared by dialysis method, and it was characterized by scanning electron microscope (SEM), rotation rheometer and thermogravimetric analyzer to observe section structure, rheological properties and thermal stability. Differential scanning calorimetry, X-ray diffraction and Fourier transform infrared spectroscopy were employed to identify the inclusion of P(NIPAM-co-AAc)-g-HA temperature sensitive material for P. americana extract, and to investigate the effect of P. americana thermosensitive hydrogel on wound healing in diabetic rats, and the rate of wound healing was calculated by Image-Pro Plus 6.0 software. Hematoxylin-eosin (HE) and Masson staining were used to observe the pathological changes of the wounds of rats in each group. Result:P(NIPAM-co-AAc)-g-HA temperature sensitive material was successfully synthesized, its LCST was between 29 ℃ and 31 ℃, it had a dense and uniform porous structure and could uniformly include P. americana extract. Pharmacodynamic studies showed that P. americana thermosensitive hydrogel group had the best effect on promoting wound healing, its infiltration degree of inflammatory cells was significantly reduced, collagen and fibroblasts arranged neatly and compactly, and the density of neovascularization was significantly increased by comparing with the model group. Conclusion:P. americana thermosensitive hydrogel can effectively promote wound healing of diabetic rats and overcome the shortage of marketed P. americana liquid preparations, this paper can provide a reference for the development of P. americana extract preparations to promote wound healing in diabetic patients.

17.
Article in Chinese | WPRIM | ID: wpr-873192

ABSTRACT

Objective:The SD rat model of hyperplasia of mammary gland(HMG) and the ultrahigh-performance liquid chromatography and mass spectrometry (UHPLC-LTQ-Orbitrap MS) technology were used to explore the pharmacological material basis of Shuangjin Sanjie granules (SJSJG) for the treatment on HMG.Method:SD rat models of HMG were administered in groups, and the nipple height and the diameter were measured; the levels of estradiol (E2), progesterone (P) and prolactin (PRL) in serum were detected, pathological examination was conducted for the hyperplasia of breast tissue. Histochemical methods were used to detect the expressions of estrogen receptor α (ERα), androgen receptor (AR), progesterone receptor (PR), tumor necrosis factor-α (TNF-α) proteins. Finally, UHPLC-LTQ-Orbitrap MS technology was used to detect the main chemical constituents of SJSJG, and the pharmacodynamic substance basis was analyzed based on the pharmacological effect.Result:The results of animal experiments showed that compared with the normal group, nipple height and diameter of the model group increased remarkably (P<0.01), serum E2 significantly increased (P<0.01). Pathological examination showed abnormal hyperplasia of breast tissue, expressions of ERα, AR, PR and TNF-α increased, compared with the model group, the nipple height and diameter of the SJSJG group decreased remarkably (P<0.01), serum E2 was decreased significantly (P<0.01), pathological examination showed weakened abnormal hyperplasia of breast tissue, ERα, AR, PR and TNF-α protein expressions were significant decreased (P<0.01). The results of basic material study showed that 85 chemical components were identified from SJSJG, including 16 alkaloids, 7 flavonoids, 15 terpenes, 9 phenolic acid compounds, 3 coumarin compounds, 10 esters and lactone compounds, 7 fatty acids compounds, 4 amino acids compounds, and 14 other types of ingredients, among them, alkaloids and terpenoids chemical drug substances were closely related.Conclusion:SJSJG can effectively improve the condition of breast hyperplasia, and its medicinal substance basis may include saikosaponin A, Saikosaponin D, verticinone, peimine.

18.
Article in Chinese | WPRIM | ID: wpr-873079

ABSTRACT

Objective::To develop high performance liquid chromatography-diode array detector (HPLC-DAD) wavelength switching for simultaneously determining the contents of inosine, loganic acid, chlorogenic acid, amygdalin, hydroxysafflor yellow A, gentiopicroside, ferulic acid and liquiritin in 15 batches of material benchmarks of Shentong Zhuyutang. Method::The quantitative analysis was carried out on a Thermo Hypersil GOLD C18 column (4.6 mm×250 mm, 5 μm) with mobile phase of acetonitrile-0.1%phosphoric acid aqueous solution for gradient elution, the flow rate was 1.0 mL·min-1, the detection wavelengths were set as 248 nm (0-11 min, inosine), 235 nm (11-14 min, loganic acid), 324 nm (14-16 min, chlorogenic acid), 220 nm (16-19 min, amygdalin and hydroxysafflor yellow A), 274 nm (19-26 min, gentiopicroside), 247 nm (26-54 min, ferulic acid and liquiritin), the column temperature was maintained at 25 ℃. According to the contents of eight active components in 15 batches of material benchmarks, orthogonal partial least squares discriminant analysis (OPLS-DA) in SIMCA 14.1 was used to evaluate the quality difference of each batch of samples. Result::Each component had good separations, the linear ranges of the above 8 components were 2.1-67.2, 1.812 5-58, 1.937 5-62, 5.212 5-166.8, 8.45-270.4, 7.075-226.4, 1.775-56.8, 3.875-124 mg·L-1, respectively (r≥0.999 6). The average recoveries of them were 99.23%, 100.09%, 99.33%, 98.85%, 99.15%, 98.75%, 99.42%, 98.96%, respectively (RSD<2%). The contents of the above eight components in 15 batches of material benchmarks were 0.183 5-0.250 3, 0.173 1-0.265 3, 0.069 5-0.169 8, 0.959 2-1.458 2, 1.905 4-2.553 3, 0.933 3-1.997 5, 0.084 6-0.143 4, 0.212 5-0.704 3 mg·g-1, respectively. Liquiritin, ferulic acid, gentiopicroside and hydroxysafflor yellow A were determined to have significant impact on the quality of different batches of material benchmarks of Shentong Zhuyutang through OPLS-DA. Conclusion::The established method for simultaneous determination of multi-components is reliable, simple and in line with the requirements of methodological verification. It is suitable for the quality control of research and development of compound preparations of Shentong Zhuyutang.

19.
Article in Chinese | WPRIM | ID: wpr-872866

ABSTRACT

Objective:To explore the efficacy and molecular mechanism of Zhizhuwan decoction and its ingredient-contained serums on the proliferation and apoptosis of rat colon interstitial cells of cajal (ICC), and make a molecule-level analysis of the possible mechanism of traditional Chinese medicine (TCM) purgation-tonifying therapy in treating slow transit constipation (STC). Method:A total of 40 rats were divided into Atractylodis Macrocephalae Rhizoma(AMR) group, Aurantii Fructus Immaturus(AFI) group, Zhizhuwan group and blank serum group on random basis, with 10 in each group. Baizhu group was given 17.7 g·kg-1·d-1 of AMR decoction by gavage, AFI group was given 8.9 g·kg-1·d-1 AFI decoction by gavage, Zhizhuwan group was given 26.4 g·kg-1·d-1 Zhizhuwan decoction by gavage, and blank serum group was given 3 mL sterile distilled water for 7 consecutive days, once a day. Drug-contained serums and blank serum were collected from blood of the above groups and diluted to 5%, 10%, 15% and 20% concentrations. Each concentration was intervened for 24 h and 48 h, and the amount and status of ICC were observed. The best intervening concentration and time for each group with cell counting kit-8 (CCK-8) were determined. Rat colon ICC was divided into blank control group, blank serum group, AMR group, AFI group and Zhizhuwan group. ICC proliferation for each group was detected with EdU, ICC apoptosis for each group was detected by flow cytometry, and expressions of X-linked inhibitor of apoptosis protein (XIAP) and proliferating cell nuclear antigen (PCNA) were detected by Western blot. Result:Compared with the normal group, the best intervention concentration for blank serum group, AMR group and AFI group was 10%, while that for Zhizhuwan group was 5%. The best intervention times for the above groups were all 24 h. No distinct difference between the effect of blank control group and blank serum group on the proliferation and apoptosis of ICC was observed. In comparison with blank control group and blank serum group, AMR group, AFI group and Zhizhuwan group showed significant changes in ICC proliferation rate (P<0.05,P<0.01). There was a greater increase in ICC proliferation rate of Zhizhuwan group than that of AMR group and Zhizhu group (P<0.05,P<0.01), with no distinct difference between the changes of ICC proliferation rates in AMR group and AFI group. There was no significant difference between the changes of ICC apoptosis rates in AMR group, AFI group and Zhizhuwan group than in blank control group and blank serum group. There were significant increases in the expressions of XIAP and PCNA in AMR group, AFI group and Zhizhuwan group than in blank control group and blank serum group (P<0.05,P<0.01), but with little difference among the three groups. Conclusion:At certain concentrations, Zhizhuwan, AFI and AMR all have the effect in improving ICC proliferation by increasing XIAP and PCNA expressions, with no evident effect on the apoptosis of ICC, based on TCM purgation-tonifying therapy, Zhizhuwan has the effect in improving ICC proliferation, with a better effect than single administration with AFI or AMR.

20.
Article in Chinese | WPRIM | ID: wpr-872726

ABSTRACT

Objective:To explore the effect of Shenqi compound on islet β-cell function in type 2 diabetic GK rats. The whole genome expression profile chip technology is used to explore the molecular mechanism of Shenqi compound regulating pancreatic islet cell function and provide theoretical basis for the prevention and treatment of type 2 diabetes with traditional Chinese medicine. Method:GK rats were fed with high-fat diet daily for 4 weeks. Rats were randomly selected from GK rats to detect random blood glucose and verified the success of type 2 diabetes model. Rats were divided into 4 groups, Wistar group, model group, Shenqi compound(1.44 g∙kg-1) group and west glenn(16 mg∙kg-1) group. After 8 weeks of gavage, the serum insulin(INS) levels were detected by enzyme-linked immunosorbent assay(ELISA). The apoptosis of islet β cells was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL)fluorescence method. Differential gene detection uses whole-genome expression profiling chip technology in each group of rat pancreatic tissues, the mRNA transcription level of key differential genes is detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR). Result:Compared with blank group, before gavage, 4 weeks, 8 weeks, GK rats have higher blood sugar in each group (P<0.01).Gavage for 4 weeks and gavage for 8 weeks, compared with model group, the blood sugar of rats in each drug intervention group was lower (P<0.01). Gavage for 8 weeks, compared with blank group, the INS level of model group was lower (P<0.01). Compared with model group, the Shenqi compound group had a higher INS level, and the sitagliptin group had a higher INS level (P<0.01). After gavage for 8 weeks, compared with the blank group, the number of pancreatic islet β-cell apoptosis in the model group was higher (P<0.05). Compared with model group, the number of pancreatic islet β cell apoptosis in the Shenqi compound group and sitagliptin group was lower (P<0.05,P<0.01). Gene chip and Real-time PCR tests both showed that phosphatidylinositol 3-kinase receptor 1(PIK3R1) was up-regulated in the Shenqi compound group/model group, and down-regulated in the sitagliptin group/model group, model group/blank group. Protein kinase B1(Akt1) was expressed in the Shenqi compound group/model The expression was up-regulated in the group, sitagliptin group/model group, and down-regulated in the model group/blank group. Conclusion:Shenqi compound which has the function of supplenmenting Qi and Yin and promoting the blood circulation, can inhibit the islet β cell apoptosis, improve islet β cell function, regulate insulin secretion, and prevent T2DM by up-regulating the expression of genes PIK3R1 and Akt1.

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