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1.
Acta Pharmaceutica Sinica ; (12): 783-792, 2022.
Article in Chinese | WPRIM | ID: wpr-922891

ABSTRACT

Molecular mass distribution of Astragalus polysaccharides is wide. Astragalus polysaccharides prepared by conventional water extraction and alcohol precipitation are mostly mixture of macromolecules. Although studies have shown that Astragalus polysaccharides have two-sided immunomodulation, the relationship between anti-inflammatory components and molecular mass distribution of Astragalus polysaccharides is not clear. Therefore, Astragalus polysaccharides were extracted by water extraction and alcohol precipitation. The relative molecular weight of them was determined by high performance gel permeation chromatography (HPGPC). Astragalus polysaccharides with different molecular weights were separated and prepared by membrane separation. RAW 264.7 cells were induced by lipopolysaccharide (LPS) to establish an inflammatory cell model in vitro and the anti-inflammatory polysaccharide were screened. The anti-inflammatory regulation mechanism of Astragalus polysaccharides was analyzed by the LC-MS/MS metabonomics technology. The results showed that APS was composed of APS-Ⅰ ( > 2 000 kDa) and APS-Ⅱ (10 kDa). APS-Ⅰ was composed of mannose, rhamnose, galacturonic acid, glucose, galactose, arabinose and the molar ratios of these monosaccharide of APS-I were 0.54∶0.26∶12.24∶17.24∶8.46∶1. APS-II was composed of rhamnose, galacturonic acid, glucose, galactose, arabinose and the molar ratios of these monosaccharide of APS-II were 0.26∶0.14∶24.04∶0.62∶1. APS-Ⅰ and APS-Ⅱ had no cell toxicity to RAW 264.7 macrophage in the range of 0-100 μg·mL-1. Compared with the model group, APS-I at a concentration of 0-100 μg·mL-1could significantly inhibit the secretion of NO and TNF-α by RAW 264.7, and can significantly promote the secretion of IL-10. APS-I had better anti-inflammatory activity than APS-II in vitro. The metabolomics results showed that 32 different metabolites were found between the model group and blank group; APS-I group can significantly callback 18 different metabolites; mainly related to arginine biosynthesis, arginine and proline metabolism, pyrimidine metabolism, citric acid cycle (TCA cycle), cysteine and methionine acid metabolism, tryptophan metabolism. This study found that APS-I had better anti-inflammatory activity than APS-II in vitro, and its mechanism may be closely related to amino acid metabolism and energy metabolism, which indicated the direction for further clarifying the pharmacodynamic material basis of Astragalus polysaccharides.

2.
Journal of Experimental Hematology ; (6): 1203-1208, 2021.
Article in Chinese | WPRIM | ID: wpr-888539

ABSTRACT

OBJECTIVE@#To investigate the relationship between the levels of ferritin, C-reactive protein (CRP), lactate dehydrogenase (LDH) and interleukin-6 (IL-6) in peripheral serum and cytokine release syndrome (CRS) in patients with relapse and/or refractory multiple myeloma (R/R MM) after receiving chimeric antigen receptor T cells (CAR-T) immunotherapy.@*METHODS@#Twenty-eight patients with R/R MM were treated with 1×10@*RESULTS@#Among the 28 patients, 27 cases (96.4%) developed CRS, 24 cases (85.7%) in 1-2 grade CRS and 3 cases (10.7%) in 3-5 grade. The severity grade of CRS of 27 patients was positively correlated with the peak values of ferritin, CRP, LDH, and IL-6 in peripheral blood (r@*CONCLUSION@#After receiving CAR-T cellular immunotherapy, the incidence of CRS in patients with R/R MM is higher, but most of them are in grade 1 or 2. The severity of CRS is positively correlated with the levels of ferritin, CRP, LDH and IL-6 in peripheral blood.


Subject(s)
Animals , Antigens, CD19 , Cytokine Release Syndrome , Humans , Immunotherapy, Adoptive , Mice , Multiple Myeloma/therapy , Neoplasm Recurrence, Local , Receptors, Chimeric Antigen
3.
Acta Pharmaceutica Sinica ; (12): 2266-2275, 2021.
Article in Chinese | WPRIM | ID: wpr-887054

ABSTRACT

We previously reported that active Astragalus polysaccharides APS-Ⅱ generate strong immune activity. Here we establish the optimal method for APS-II acid degradation. After preliminary structural studies and separation and preparation of the degradation products, the oligosaccharide active center with the strongest immune activity was identified by in vitro immune cell culture experiments. The optimum acid degradation conditions for APS-II were determined by a single factor experiment and an orthogonal experiment. Astragalus oligosaccharides prepared under the optimal conditions were subjected to structural analysis by hydrophilic interaction chromatography - electrospray ionization source - high resolution time-of-flight mass spectrometry. The products were separated and oligosaccharide fragments with different degrees of polymerization were isolated by preparative purification chromatography. Finally, fragments of the immunologically active centers were identified by in vitro immune cell cultures from multiple perspectives. The results show that the optimal acid hydrolysis conditions for APS-Ⅱ are hydrolysis temperature 80 ℃, trifluoroacetic acid concentration 1.0 mol·L-1, hydrolysis time 1 h. The degradation conditions have good repeatability. The degradation product is a six-carbon aldehyde glycan structure with the main chain 1→4 connected. The immune activity screening experiment for six oligosaccharide fragments showed that larger molecular weight oligosaccharides have stronger immune-promoting effects. It is speculated that the immunologically active center of Astragalus oligosaccharide is located in the sugar chain of DP9-DP19. The animal welfare and the experimental process in this study follow the requirements of the Animal Ethics Committee of Shanxi University. This result suggests a foundation for the structural characterization and structure-activity relationship research of Astragalus oligosaccharides, and may promote the development of Astragalus oligosaccharide drugs.

4.
Acta Pharmaceutica Sinica ; (12): 1936-1944, 2021.
Article in Chinese | WPRIM | ID: wpr-887009

ABSTRACT

italic>Astragalus polysaccharides are the main immunomodulatory substances in Astragali Radix. The structure of polysaccharides is difficult to accurately determine, which limits the in-depth study of the molecular mechanism of Astragalus polysaccharides in vivo. "Polysaccharide receptor theory" believes that there are one or more oligosaccharide fragment "active centers" in immunologically active polysaccharide molecules. Therefore, the degradation of Astragalus polysaccharides into oligosaccharides and the study of the active centers of polysaccharides at the oligosaccharide level provide new ideas in the study of the structure and mechanism of Astragalus polysaccharides. This article adopts endo-α-1,4-glucanase enzymatic hydrolysis, and determines the best degradation conditions through single factor test and orthogonal test to degrade the immunologically active polysaccharide APS-Ⅱ (10 kDa component) into oligomers with different degrees of polymerization. Then through the preparation of polyacrylamide gel chromatography and specific immune and non-specific immune cell tests, the immune activity screening of different oligosaccharide components is carried out. The animal welfare and the experimental process in this study follow the requirements of the Animal Ethics Committee of Shanxi University. The results showed that compared with the immunologically active polysaccharide APS-Ⅱ, different oligosaccharide components have obvious differences in different immunological activities. This paper studies the different immunological activities of Astragalus polysaccharides at the level of oligosaccharides, laying a foundation for further elucidating the structure and function of Astragalus polysaccharides, enriching the theory of polysaccharide receptors, and providing new ideas for the development of Astragalus polysaccharides.

5.
Article in Chinese | WPRIM | ID: wpr-880150

ABSTRACT

OBJECTIVE@#To investigate the effects of autophagy inhibitor ROC-325 and its combination with bortezomib on the proliferation, apoptosis and autophagy of multiple myeloma cell lines.@*METHODS@#Multiple myeloma cells were treated with ROC-325 at different concentration. The cell proliferation was detected by CCK-8. Apoptosis was determined by Caspase-3/7 and Caspase-9 activity assays. Autophagy was detected by monodansylcadaverine staining. The apoptosis-related proteins (PARP and Caspase-3) and autophagy-related proteins (P62, Beclin-1, and LC3A/B) were analyzed by Western blot. The combined effect with bortezomib on bortezomib-resistant cell line was detected by CCK-8.@*RESULTS@#ROC-325 inhibited the proliferation of RPMI 8226, RPMI 8226-BTZ100, U266 and IM9 cells in a dose-dependent manner (r=-0.8275, r=-0.9079, r=-0.9422, r=-0.9305), the 72 h IC@*CONCLUSION@#ROC-325 can inhibit the proliferation, induce the apoptosis of myeloma cells through the mitochondrial pathway, inhibit the autophagy of myeloma cells by affecting the fusion of autophagosomes and lysosomes, and overcome bortezomib resistance by the combination of ROC-325 with bortezomib.


Subject(s)
Apoptosis , Autophagy , Bortezomib/pharmacology , Cell Line, Tumor , Cell Proliferation , Humans , Hydroxychloroquine/analogs & derivatives , Multiple Myeloma
6.
Article in Chinese | WPRIM | ID: wpr-906441

ABSTRACT

Objective:To elucidate the potential molecular markers and drug-compound-target mechanism of Epimedii Folium intervention on breast cancer stem cells(BCSCs) through chip analysis combined with network pharmacology and experimental validation. Method:Relevant drug information was retrieved in Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) to obtain the active components and potential targets of Epimedii Folium. "Breast Cancer Stem Cells" were searched in Gene Expression Omnibus(GEO)database,and GSE98239 chip data were obtained through analysis and screening. Then GEO2R online analysis tool was used to obtain the differential genes to draw differential gene heat map and volcano map. The differential gene network map of Epimedii Folium intervention for breast cancer stem cells was constructed by Cytoscape 3.8.0,and Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis of drug and disease genes were performed. Human breast cancer MDA-MB-231 cells were divided into 20%,40%,60% Epimedii Folium drug-containing serum group and control group. Cell counting kit-8(CCK-8),and Western blot were used to detect the effect of Epimedii Folium drug-containing serum intervention on cell activity and target protein expression in breast cancer cells. Result:Twenty-three active components including flavones,sterols,alkaloids and sesquiterpenoids were obtained from Epimedii Folium. It was found that Epimedii Folium interacted with B-cell lymphoma-2-like protein 1(BCL2L1),matrix metallopeptidase 2(MMP2),prostaglandin-endoperoxide synthase 2(PTGS2),vascular endothelial growth factor A(VEGFA),transforming growth factor beta receptor 1(TGFBR1) and other pivotal genes in breast cancer stem cells,participated in the induction of new angiogenesis and cell migration,enabled the continuous self-renewal of BCSCs,decreased apoptosis and cell migration,thus promoting the recurrence and metastasis of breast cancer. KEGG results showed that Epimedii Folium intervened in multiple differential expressed genes(DEGs)of transforming growth factor-<italic>β</italic>(TGF-<italic>β</italic>),vascular endothelial growth factor(VEGF),phosphoinositide 3kinase/protein kenase B(PI3K/Akt),mitogen-activated protein kinese(MAPK)and mammalian target of rapamycin(mTOR)subpathways in cancer signaling pathways to exert its efficacy in intervening breast cancer stem cells. Experiments showed that the survival rate of breast cancer cells was significantly reduced and the expression levels of TGFBR1 and Smad2 in breast cancer cells significantly decreased after the intervention of Epimedii Folium drug-containing serum(<italic>P</italic><0.01). Conclusion:Several components in different concentrations of drug-containing serum of Epimedii Folium can synergistically act on target differentially expressed genes of breast cancer stem cells,and inhibit the proliferation of breast cancer cells by down-regulating the expression levels of TGFBR1,a key molecule in the TGF-<italic>β</italic> pathway,and Smad2,a downstream signal.

7.
Article in Chinese | WPRIM | ID: wpr-921754

ABSTRACT

To investigate the potential molecular markers and drug-compound-target mechanism of Mahuang Shengma Decoction(MHSM) in the intervention of acute lung injury(ALI) by network pharmacology and experimental verification. Databases such as TCMSP, TCMIO, and STITCH were used to predict the possible targets of MHSM components and OMIM and Gene Cards were employed to obtain ALI targets. The common differentially expressed genes(DEGs) were therefore obtained. The network diagram of DEGs of MHSM intervention in ALI was constructed by Cytoscape 3. 8. 0, followed by Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses of target genes. The ALI model was induced by abdominal injection of lipopolysaccharide(LPS) in mice. Bronchoalveolar lavage fluid(BALF) was collected for the detection of inflammatory factors. Pathological sectioning and RT-PCR experiments were performed to verify the therapeutic efficacy of MHSM on ALI. A total of 494 common targets of MHSM and ALI were obtained. Among the top 20 key active compounds of MHSM, 14 from Ephedrae Herba were found to be reacted with pivotal genes of ALI [such as tumor necrosis factor(TNF), tumor protein 53(TP53), interleukin 6(IL6), Toll-like receptor 4(TLR4), and nuclear factor-κB(NF-κB)/p65(RELA)], causing an uncontrolled inflammatory response with activated cascade amplification. Pathway analysis revealed that the mechanism of MHSM in the treatment of ALI mainly involved AGE-RAGE, cancer pathways, PI3 K-AKT signaling pathway, and NF-κB signaling pathway. The findings demonstrated that MHSM could dwindle the content of s RAGE, IL-6, and TNF-α in the BALF of ALI mice, relieve the infiltration of inflammatory cells in the lungs, inhibit alveolar wall thickening, reduce the acute inflammation-induced pulmonary congestion and hemorrhage, and counteract transcriptional activities of Ager-RAGE and NF-κB p65. MHSM could also synergically act on the target DEGs of ALI and alleviate pulmonary pathological injury and inflammatory response, which might be achieved by inhibiting the expression of the key gene Ager-RAGE in RAGE/NF-κB signaling pathway and downstream signal NF-κB p65.


Subject(s)
Acute Lung Injury/genetics , Animals , Drugs, Chinese Herbal/pharmacology , Lipopolysaccharides , Lung/metabolism , Mice , NF-kappa B/metabolism , Receptor for Advanced Glycation End Products/metabolism , Signal Transduction
8.
Acta Pharmaceutica Sinica ; (12): 843-853, 2020.
Article in Chinese | WPRIM | ID: wpr-821678

ABSTRACT

Oligosaccharides play important roles in many biological processes and have great medicinal potential. The structure-activity study of oligosaccharides is complicated by the diversity and complexity of their structures. This review summarizes the state of oligosaccharide structural analysis in recent years. The ionization methods and dissociation rule of acid oligosaccharides, neutral oligosaccharides and glycopeptides in mass spectrometry are discussed and the methods of extraction and purification of oligosaccharides, the suitable mass spectrometry models for different types of oligosaccharides, and the advantages and disadvantages of different mass spectrometry models are introduced. We believe that this review will be helpful for the further investigation of these important biological substances.

9.
Acta Pharmaceutica Sinica ; (12): 979-986, 2020.
Article in Chinese | WPRIM | ID: wpr-821674

ABSTRACT

"Beany flavor" is one of the important authentic characteristics of Astragali Radix. It is important to clarify the material basis of "Beany flavor" in the characterization of authentic medicinal materials and establishing quality control methods for authentic medicinal materials. At present, the analysis of volatile components in different habitats has been reported, but systematic comprehensive research has not been conducted. The SPME-GC-MS technique was used to analyze the volatile components of Astragali Radix from three producing areas: Shanxi (wild-simulated Astragali Radix), Gansu (cultivated Astragali Radix) and Inner Mongolia (cultivated Astragali Radix). Combined with the method of multivariate statistical analysis, the difference of volatile components of Astragali Radix from the three different producing areas is discussed. Multivariate statistical analysis and evaluation was conducted through principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). Seventy-six volatile components of Astragali Radix from the three producing areas were identified by SPME-GC-MS. PCA model results showed that the volatile components correlated with their producing areas. PLS-DA demonstrated that the volatile components of Astragali Radix samples from different producing areas were significantly different. Six volatile substances including hexanoic acid, hexanal, 2-butyl-2-octenal, 1-hexanol, benzyl alcohol and 2-butylfuran were quantified. The ratio of hexanal to hexanoic acid in Astragali Radix from different areas and cultivation methods was significantly different: the ratio of hexanal: hexanoic acid in wild-simulated Astragali Radix of Shanxi was 7.8∶1, while the ratio of hexanal: hexanoic acid in cultivated Astragali Radix of Inner Mongolia and Gansu was 2.3∶1 and 0.96∶1, respectively. The volatile components identified in this study provide a foundation for continued research of and quality control of Astragali Radix.

10.
Acta Pharmaceutica Sinica ; (12): 930-940, 2020.
Article in Chinese | WPRIM | ID: wpr-821666

ABSTRACT

The aim of this study was to analyze the anti-cancer effect and mechanism of action of the flavonoids of Astragalus membranaceus (TFA) when combined with cisplatin on Lewis lung carcinoma-bearing mice. This animal experiment was approved by the Committee of the Ethics of Animal Experiment of Shanxi University (SXULL2018012). Pharmacological indices such as tumor weight, tumor volume growth curves, inhibition rate and organ indices showed that the TFA could reduce toxicity and enhance the efficacy of cisplatin. The target of TFA was predicted by network pharmacology analysis and the result showed that calycosin-7-O-β-D-glucoside might be the main active compound responsible for the anticancer effect of TFA. TRP53 (cellular tumor antigen p53), RAC1 (Ras-related C3 botulinum toxin substrate 1), ERBB2 (receptor tyrosine-protein kinase erbB-2), VEGFA (vascular endothelial growth factor A) and STAT3 (signal transducer and activator of transcription 3) may be associated with TFA in enhancing efficacy and reducing the toxicity of cisplatin. The IL-6 content in serum and expression levels of STAT3 and p53 in tumor tissues suggested that TFA may inhibit tumor growth through the IL-6/STAT3 pathway; UPLC-MS-based serum metabolomic analysis suggested that the metabolic pathways related to lung cancer include sphingolipid metabolism, retinol metabolism, glycerophospholipid metabolism, primary bile acid biosynthesis, and the TFA-regulated corresponding pathway of bile acid biosynthesis. In this study, the anti-cancer effect and mechanism of action of TFA combined with cisplatin on Lewis lung carcinoma-bearing mice was analyzed by the combination of various techniques, which lay a foundation for further development of anticancer drugs.

11.
Acta Pharmaceutica Sinica ; (12): 2968-2975, 2020.
Article in Chinese | WPRIM | ID: wpr-862275

ABSTRACT

Characterization of the polysaccharides and monosaccharides of Bupleurum chinense was undertaken to identify differences in the Bupleurum chinense's sugar profiles, so as to provide a basis for the identification of different varieties. High performance liquid chromatography (HPLC) was used to generate chromatograms of the total polysaccharides of Bupleurum using an Evaporation Light Detector (ELSD), and a monosaccharide chromatogram was generated using a UV-detector (UV) following polysaccharide derivatization. The data were analyzed using SIMCA software and SPSS software to distinguish different varieties of Bupleurum. The results show that the yield of polysaccharides from Bupleurum falcatum is the highest, while the yield of polysaccharides from Bupleurum chinense is the lowest. The polysaccharide spectrum shows that the molecular weights of the polysaccharides in different Bupleurum differ, and their percentages of the total peak area are also different. The four Bupleurum polysaccharides are composed of mannose, glucuronic acid, rhamnose, galacturonic acid, glucose, galactose, and arabinose, but differ in length. The ratio of glucose to arabinose in Bupleurum chinense, Bupleurum scorzonerifolium, Bupleurum falcatum and Bupleurum marginatum var. stenophyllum is: 3.0-4.0, 5.5-7.0, 12.0-17.0, 9.0-12.0. In this study, a sugar profile technique was developed to provide a new method for the identification of different varieties of Bupleurum.

12.
Article in Chinese | WPRIM | ID: wpr-872926

ABSTRACT

Objective:The chemical differences of Lycii Fructus samples from Qinghai, Ningxia, Gansu, Xinjiang and Inner Mongolia provinces were compared based on proton nuclear magnetic resonance (1H-NMR) plant metabolomics. Method:A total of 97 Lycii Fructus samples from five provinces were collected, including 61 samples in Qinghai, and extracted by 50% methanol for detecting. 1H-NMR spectra were obtained and compared by multivariate statistical analysis for investigating the chemical differences of samples from Qinghai and other production areas. And the content of Lycii Fructus polysaccharides in all samples was determined with the wavelength of 490 nm (calculated by anhydrous glucose). Result:A total of 32 chemical components were detected in the Lycii Fructus extract by 1H-NMR. The multivariate statistical analysis revealed that there was no significant difference among the samples from five provinces. The difference between Lycii Fructus from Qinghai and Ningxia, as well as the samples among the six regions of Qinghai province were relatively small. The similarity values of the majority of samples were >0.85. Univariate analysis showed that no significant difference was observed for the most metabolites in Lycii Fructus collected from five provinces, except for sucrose, glucose, proline and so on. There was no significant difference in the content of Lycii Fructus polysaccharides between Qinghai and other provinces. And the correlation coefficient between the content of Lycii Fructus polysaccharides and the small molecular compounds identified by 1H-NMR was -0.2-0.4. Conclusion:In this study, chemical characteristics of Lycii Fructus in Qinghai province are analyzed from the holistic view by 1H-NMR plant metabolomics, in combination of polysaccharide determination, and the results show that there is no significant difference between samples from Qinghai and other four provinces. The quality evaluation method based on 1H-NMR established in this study can provide scientific basis for improving quality control level and selecting planting areas of Lycii Fructus.

13.
Acta Pharmaceutica Sinica ; (12): 2414-2420, 2020.
Article in Chinese | WPRIM | ID: wpr-829376

ABSTRACT

Asthma is a chronic inflammatory disease involving eosinophils, mast cells, lymphocytes, neutrophils and other inflammatory cells and immune components. Chinese medicine has unique advantages in the treatment of asthma. In this study an ovalbumin (OVA)-induced asthma model in rats was used to verify the effect of the total sesquiterpenoids of Flos Farfarae (F.F.S) on asthma. The results show that the F.F.S has anti-asthmatic effects. Metabolomic analysis showed that 25 metabolites were changed in asthmatic rats, and 18 of them could be attributed to F.F.S. F.F.S. can block the over-activation of p65 NF-κB, preventing the transmission of pro-inflammatory factor signals to cells. F.F.S. thus could reduce the over-activation of inflammatory cells, inhibiting the secretion of inflammatory factors and further alleviating asthma inflammation. This study lays a foundation for further new drug research. The mechanism of F.F.S. on asthma needs further investigation.

14.
Article in Chinese | WPRIM | ID: wpr-829048

ABSTRACT

OBJECTIVE@#To study the expression of multiple negative costimulatory molecules on peripheral blood T cells in patients with acute myeloid leukemia (AML) and its affection on prognosis.@*METHODS@#The peripheral blood samples from patients with newly diagnosed AML, complete remission (CR), and no-remission (NR) were collected, the expression levels PD-1、VISTA and TIM-3 in CD4 and CD8 T cells were detected by flow cytometry , and the clinical data of patients were analyzed.@*RESULTS@#The expression levels of PD-1、VISTA and TIM-3 of CD4 and CD8 T cells in the newly diagnosed AML patients were significantly higher than those in control group (P<0.05). The expression levels of PD-1、TIM-3 and VISTA of CD4 and CD8 T cells in the CR group were significantly lower than those in newly diagnosed and the NR group (P<0.05). The TIM-3 expression level positively correlated with VISTA expression level of CD4 and CD8 T cells in newly diagnosed AML patients (r=0.85 and 0.73). The VISTA and PD-1 expression level of CD4 T cells in newly diagnosed AML, NR after first induction chemotherapy and high risk patients significantly increased (P<0.05), the TIM-3 expression level of CD8 T cells in high risk group significantly increased (P<0.05), and the VISTA expression level of CD8 T cells in CBFβ-MYH11 mutation-positive group significantly decreased (P<0.05).@*CONCLUSION@#The expression of PD-1、TIM-3 and VISTA in AML peripheral blood T cells may be involved in the immune escape of AML and can be the targets of treatment for acute myeloid leukemia patients.


Subject(s)
B7 Antigens , CD8-Positive T-Lymphocytes , Flow Cytometry , Hepatitis A Virus Cellular Receptor 2 , Humans , Leukemia, Myeloid, Acute , Programmed Cell Death 1 Receptor
15.
Article in Chinese | WPRIM | ID: wpr-828682

ABSTRACT

OBJECTIVE@#To study the reference ranges of platelet and related parameters within 24 hours after birth in preterm infants with different gestational ages.@*METHODS@#According to the inclusion and exclusion criteria, a retrospective analysis was performed for the chart review data of 1 070 preterm infants with a gestational age of 23-36 weeks who were admitted to the neonatal intensive care unit from January to December in 2018. The reference ranges of platelet parameters were calculated for the preterm infants within 24 hours after birth.@*RESULTS@#There were no significant differences in platelet count (PLT) and plateletcrit (PCT) among the preterm infants with different gestational ages (P>0.05). The late preterm infants (34-36 weeks; n=667) had significantly lower mean platelet volume (MPV) and platelet distribution width (PDW) than the extremely preterm infants (23-27 weeks; n=36) and the early preterm infants (28-33 weeks; n=367) (P0.05). The reference ranges of platelet parameters in preterm infants were calculated based on gestational age. The reference ranges of PLT and PCT were (92-376)×10/L and 0.1%-0.394% respectively, for the preterm infants with a gestational age of 23-36 weeks. The reference ranges of MPV and PDW were 9.208-12.172 fl and 8.390%-16.407% respectively, for the preterm infants with a gestational age of 23-36 weeks; the reference ranges of MPV and PDW were 9.19-11.95 fl and 9.046%-15.116% respectively, for the preterm infants with a gestational age of 34-36 weeks.@*CONCLUSIONS@#The MPV and PDW of preterm infants with different gestational age are different within 24 hours after birth, and it is more helpful for clinical practice to formulate the reference range of MPV and PDW according to gestational age.


Subject(s)
Blood Platelets , Gestational Age , Humans , Infant, Newborn , Mean Platelet Volume , Reference Values , Retrospective Studies
16.
Article in Chinese | WPRIM | ID: wpr-828425

ABSTRACT

The present study was performed to establish the UPLC fingerprints of Bolbostemmatis Rhizoma and determine the contents of three saponins by quantitative analysis of multi-components by single marker(QAMS), and provide basis for quality evaluation of Bolbostemmatis Rhizoma. The analysis was carried out on an analytical column of Waters Cortecs T3(2.1 mm×100 mm,1.6 μm)with gradient elution by acetonitrile-0.1% phosphoric acid solution, at a flow rate of 0.3 mL·min~(-1). The detection wavelength was 203 nm, the column temperature was 30 ℃ and the injection volume was 1 μL. The UPLC fingerprints of Bolbostemmatis Rhizoma were established and evaluated by similarity calculation, cluster analysis and principal component analysis. The relative calibration factors of toberoside B and toberoside C were determined with toberoside A as internal reference. The content was calculated by relative calibration factors to develop a method of QAMS. Comparing the results of QAMS with those of ESM, the accuracy and feasibility of one-eva-luation and multi-evaluation can be determined. RESULTS:: showed that the fingerprints of 19 batches of Bolbostemmatis Rhizoma have four common peaks with similarities ranging from 0.754 to 1.000. Cluster analysis and principal component analysis classified 19 batches of Bolbostemmatis Rhizoma into three categories, which was consistent with the similarity evaluation results. The relative deviation between the content of tubeicosides B and C in 19 batches of Bolbostemmatis Rhizoma determined by QAMS and ESM is less than 5.0%, indicating that there was no significant difference between the two methods. Therefore, the UPLC fingerprints combined with QAMS and similarity evaluation can be effectively used to evaluate the quality of Bolbostemmatis Rhizoma.


Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Principal Component Analysis , Quality Control , Rhizome
17.
Article in Chinese | WPRIM | ID: wpr-827196

ABSTRACT

OBJECTIVE@#To explore the expression and clinical significance of EZH2 in DLBCL patients accompanied by HBV infection.@*METHODS@#The clinicopathological data of 59 patients with DLBCL accompanied by HBV infection in our hospital from February 2015 to October 2017 were analyzed retrospectively. The patients were divided into HBV negative and HBV positive groups by serological testing before surgery. The expression of EZH2 was detected by immumohistochemical staining, and the clinicopathological characteristics and survival were analyzed and compared between these two groups.@*RESULTS@#There were 30 patients (50.8%) in the HBV negative group and 29 patients (49.2%)in the HBV positive group. The differences of age, LDH level and IPI score between two groups were statistically significant (P<0.05). The expression of EZH2 in HBV- positive group was significantly higher than that in the HBV- negative group (P<0.05), where the expression of EZH2 correlated with the expression of the BCL-6 (r=0.282, P<0.05), especially in the GCB-DLBCL (r=0.549, P<0.05). PFS was not significantly different between two groups of HBV (P>0.05), while the PFS in the R-CHOP regimen group was higher than that in the CHOP regimen group (P<0.05). COX multivariate analysis showed that both the chemotherapy regimen without R and the increased level of LDH were the risk factors affecting the prognosis of DLBCL patients (P<0.05).@*CONCLUSION@#EZH2 highly expresses in HBV positive group, suggesting that the significance of EZH2 in DLBCL with HBV infection is worth further explore.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols , Cyclophosphamide , Doxorubicin , Enhancer of Zeste Homolog 2 Protein , Genetics , Hepatitis B , Hepatitis B virus , Humans , Lymphoma, Large B-Cell, Diffuse , Genetics , Prognosis , Retrospective Studies , Rituximab , Vincristine
18.
Journal of Experimental Hematology ; (6): 1228-1233, 2020.
Article in Chinese | WPRIM | ID: wpr-827135

ABSTRACT

OBJECTIVE@#To investigate the expression and clinical significant of VCAN and its related molecules in patients with MM.@*METHODS@#Ficoll density gradient centrifugation method was used to speared the bone marrow mononuclear cell in 25 cases of MM before and after treatment, the relative mRNA expression of VCAN and their related molecules (FAK, FN, MK, and HAS) in bone marrow was detected by real-time quantitative PCR, and their protein expression was determined by Western bolt.@*RESULTS@#The expression of VCAN, FK and FN in the effective group after treatment was significantly lower than that before treatment (P<0.05), however, the expression of MK and HAS showed no statistically significantly different before and after treatment (P<0.05). The expression of VCAN of patients in non remission group was significantly higher than that in control group (P<0.05). The expression of FAK and FN of patients in no remission group was significant increased as compared with the patients in newly diagnosed group (P<0.05). The relative expression of VCAN mRNA in the patients at 3rd stage was significantly higher than those at the 1st stage (P<0.05) and control group but showed no significant difference to the patients at 2nd stage (P<0.05). The expression of VCAN and its related proteins (FAK, MK, FN) showed positively correlation in bone marrow mononuclear cells of MM patients (P<0.05). The correlation between VCAN and HAS was not statistically significant (r=0.259,P>0.05). Survival analysis showed that the relative expression of VCAN mRNA was associated with OS (P=0.049) and PFS (P=0.041) in MM patients.@*CONCLUSION@#VCAN and its related molecules are highly expressed in MM patients; VCAN may act as potential biomarker in the development of multiple myeloma.


Subject(s)
Bone Marrow , Humans , Multiple Myeloma , RNA, Messenger , Versicans
19.
Article in Chinese | WPRIM | ID: wpr-781461

ABSTRACT

OBJECTIVE@#To study the level and clinical value of Th17 cell subset in the peripheral blood of the patients with aplastic anemia (AA).@*METHODS@#Eighty-five patients with aplastic anemia in our hospital from May 2017 to February 2018 were selected. Among them, 51 patients with poor curative effect were enrolled in group A. and the 34 patients with good curative effect were enrolled in group B, the 35 healthy persons were selected as controls. The levels of serum IL-17, IL-6, IL-4 and IFN-β were analyzed by ELISA; the levels of peripheral blood Th17 cell subset were analyzed by flow cytometry; the expression levels of RORγt and STAT3 mRNA in lymphocytes were analyzed by RT-PCR; the expression levels of RORγt and STAT3 protein in lymphocytes were analyzed by Western blot.@*RESULTS@#There was no significant difference in sex, age, WBC count and complications between group A and group B (P>0.05). Non-severe aplastic anemia (NSAA) in group B accounted for 23 cases, and the NSAA ratio (23/34) was significantly higher than that in group A (20/51) (P<0.05). The Hb level (86.25±7.9 g/L) and Plt count (54.7 ± 6.3×10/L) in group B were significantly higher than those in group A (P<0.05). ELISA showed that the levels of IL-17 and IL-6 in group A were significantly higher than those in control group (P<0.05); the levels of IL-17 and IL-6 in group B were significantly lower than those in group A (P<0.05); the levels of IL-4 and IFN-β in group A were significantly lower than those in control group (P<0.05); the levels of IL-4 and IFN-βin group B were significantly higher than those in group A (P<0.05). Flow cytometry showed that the proportion of Th17 cells in peripheral blood of group A was higher than that of group B (P<0.05). RT-PCR showed that the levels of RORγt and STAT3 mRNA in group A were significantly higher than those in control group (P<0.05), and the mRNA levels of RORγt and STAT3 in group B were significantly lower than those in group A (P<0.05). Western blot showed that the protein levels of RORγt and STAT3 in group A were significantly higher than those in control group (P<0.05), and the protein levels of RORγt and STAT3 in group B were significantly lower than those in group A (P<0.05).@*CONCLUSION@#The Th17 cells in peripheral blood of AA patients increase, and the inflammatory reaction in the patients are enhanced. It may be related with the rise of RORγt and STAT3 gene expression, which provides a research direction for clinical treatment of AA.

20.
Article in Chinese | WPRIM | ID: wpr-771924

ABSTRACT

OBJECTIVE@#To explore the role of neutrophil-to-lymphocyte ratio(NLR) in patients with newly diagnosed essential thrombocythemia(ET) and its relationship with thrombotic events.@*METHODS@#The clinical and follow-up data of 150 ET patients were retrospectively analyzed. The risk factors of thrombotic events and role of NLR by statistical methods.@*RESULTS@#Age (P<0.01) and JAK2V617F mutation (P<0.01) were the independent risk factors for thrombotic events at diagnosis; WBC count (P<0.05), NLR (P<0.01), age (P<0.05) and thrombosis history at diagnosis (P<0.05) were independent risk factors for future thrombotic events. The ROC curve showed that NLR for prediction of future thrombotic events was suprior to other risk factors. The Kaplan-Meier analysis showed that the progress-free survival time in thrombotic events patients with higher NLR (median survival 22.3 months, 95% CI:17.8-26.8) was significantly shorter than that of patients with lower NLR (median survival 55.5 months, 95% CI:53.4-57.5) . After follow-up 60 months, the thrombosis progress-free survival in lower NLR patients reached 97.4%, while that in the patients with higher NLR was rate 46.7%.@*CONCLUSION@#NLR at diagnosis is a better predictive parameter for future thrombotic events than other clinical parameters in ET patients, but without corralations with thrombosis at diagnosis.


Subject(s)
Humans , Lymphocyte Count , Lymphocytes , Neutrophils , Prognosis , Retrospective Studies , Thrombocythemia, Essential , Thrombosis
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