1. Mechanism of ginsenoside CK inhibiting proliferation of human colon cancer SW480 cells
Chinese Traditional and Herbal Drugs 2020;51(6):1567-1574
Objective: To study the effects of ginsenoside CK on proliferation and apoptosis of human colon cancer cell line SW480, and further explore the mechanism. Methods: Cell viability was measured by CCK-8 assay. Cell cycle, apoptosis, reactive oxygen species (ROS) levels and changes in mitochondrial membrane potential were measured by flow cytometry. Hoechst staining further detected apoptosis. Western blotting was used to detect the release of cytochrome C and the expression of apoptosis-related proteins such as Bcl-2, Bax and cleaved Caspase-3. Results: Ginsenoside CK had a significant inhibitory effect on the proliferation of human colon cancer cell line SW480. Ginsenoside CK induced SW480 cells arrest in G0/G1 phase, promoted early apoptotic cells, significantly increased intracellular ROS levels and reduced the MMP level. Ginsenoside CK promoted the expression of Bax and cleaved-Caspase-3 and inhibited the expression of Bcl-2. In addition, ginsenoside CK released a large amount of cytochrome C in SW480 cells. Conclusion: Ginsenoside CK has a significant inhibitory effect on the proliferation of human colon cancer cell line SW480. The mechanism may be through the promotion of mitochondrial superoxide elevation, resulting in a significant increase in intracellular ROS levels and a significant decrease in MMP level, further leading to the release of cytochrome C, the up-regulated expression of Bax, the down-regulated expression of Bcl-2, and ultimately leading to apoptosis of cells.
2.Relationship of endothelial nitric oxide synthase gene polymorphism, the 27-bp repeat in intron 4 with essential hypertension of the northern Han nationality in China
Xiaoyun ZHAO ; Xue GUO ; Changchun QIU ; Donghui ZHANG ; Yepu SU
Chinese Journal of Rehabilitation Theory and Practice 2005;11(6):422-424
ObjectiveTo investigate the relationship of the polymorphism of endothelial nitric oxide synthase (eNOS), the 27-bp variable number of tandem repeats (VNTR) in intron 4 with essential hypertension (EH) of the northern Han nationality in China.MethodsGenotypes, the level of plasma nitric oxide metabolites (NOx) and the activity of nitric oxide synthase (NOS) of 207 EH subjects and 231 healthy subjects were measured by polymerase chain-reaction (PCR).ResultsThe frequencies of ecNOS4a/a,ecNOS4b/a, and ecNOS4 b/b in the healthy group were 0.43%, 13.42% and 86.15% respectively. The frequency of the b allele was 92.86%, and the frequency of the a allele was 7.14%. While the frequencies of ecNOS4 a/a, ecNOS4 a/b,and ecNOS4 b/b in the EH group were 0.49%, 19.32% and 80.19% respectively. The frequency of the a allele in EH group (n=42, 10.15%) was significantly higher than that in the healthy group (n=33, 7.14%)(P<0.05). The plasma NOx level of the EH group was 70.04±14.68 mol/L, and significantly lower than that 84.09±27.27 mol/L in the healthy group (P<0.05). Similarly, both the plasma TNOS and iNOS activities of the EH group were 35.49±12.8 U/ml and 14.92±7.93 U/ml, and markedly lower than that 41.47±13.2 U/ml and 10.11±6.21U/ml in the healthy group (P<0.05). But the activities of eNOS in the EH group and healthy group were not significantly different (P>0.05).ConclusionThe variations of ecNOS4 gene locus may be responsible for the decrement of plasma NOx, both plasma NOx level and activity of NOS decreases in EH patients, so it may be a genetic susceptibility marker for EH of the Han nationality in China.
3.Effects of Exogenous Fusarium oxysporum and Trichoderma viride on Soil Fungal Community of Panax ginseng
Jing FANG ; Li-li WENG ; Chun-ping XIAO ; Di LI ; Xue-qing YANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(6):114-123
ObjectiveTo investigate the effects of exogenous Fusarium oxysporum and Trichoderma viride on the diversity of soil fungal community and carbon metabolic function of cultivated Panax ginseng. MethodIllumina HiSeq 2500 high-throughput sequencing combined with Biolog-ECO was used to analyze the species diversity and functional diversity of soil fungal communities in P. ginseng soil under different exogenous treatments. ResultThe results of high-throughput sequencing showed that the number and species of microorganisms in the soil were significantly changed after exogenous microorganisms were added. The soil fungi with relative abundance greater than 1% included Mortierella sp.,Fusarium sp.,Humicola sp.,and Simplicillium sp. Mortierella sp. in each treatment group significantly increased. Humicola sp. and Simplicillium sp. could be induced to increase by exogenous addition of F. oxysporum,while T. viride at a high concentration could significantly inhibit the growth of F. oxysporum. As revealed by Biolog and principal component analysis (PCA),the average well color development (AWCD) in the high-dose T. viride group (MG) was significantly higher than that in the control group (QS)and the low-dose F. oxysporum group(LD). The utilization abilities for amino acids,carboxylic acids,polymers, and amines were enhanced in the MG group,but the microbial metabolic activity was reduced in the high-dose F. oxysporum group (LG). There was no significant increase in the utilization of phenolic acids by soil microorganisms in both groups. ConclusionExogenous addition of F. oxysporum can lead to the growth and reproduction of other pathogenic fungi. Exogenous addition of T. viride can enhance the soil fungal community structure and metabolic diversity,inhibit the proliferation of F. oxysporum,and improve the soil microbial environment of cultivated P. ginseng.
4.Wutou Chishizhi Wan Regulates Autophagy and PI3K/Akt/GSK-3β Signaling Pathway in Rats with Myocardial Ischemia-reperfusion Injury
Yu-xue XIAO ; Xiao-mei SHI ; Lu-lu XIE ; Xin-he WANG ; Song WANG ; Zhao-peng ZHANG ; Chun-yu LIAN ; Jun-peng GUO ; Hong-yan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(11):26-32
ObjectiveTo explore the mechanism of Wutou Chishizhi Wan in regulating autophagy and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) signaling pathway in rats with myocardial ischemia-reperfusion injury (MIRI). MethodSixty male SD rats were randomly assigned into the normal group (normal saline), model group (normal saline), positive control (trimetazidine, 5.4 mg·kg-1) group, and low-, medium-, and high-dose (1.63, 4.9, 14.7 g·kg-1, respectively) Wutou Chishizhi Wan groups, with 10 rats in each group. The rats in other groups except the normal group underwent left anterior descending coronary artery ligation for modeling. Electrocardiogram was employed to detect the ST-segment elevation to evaluate the modeling. Hematoxylin-eosin (HE) staining was performed to reveal the damage of myocardial tissue. The levels of aspartate aminotransferase (AST) and creatine kinase (CK) were determined by colorimetry, and those of cardiac troponin T (cTnT) and myoglobin (MYO) by enzyme-linked immunosorbent assay (ELISA). Western blot was carried out to determine the protein levels of microtubule-associated proteins 1 light chain 3 (LC3), autophagy-related gene Beclin-1, PI3K, Akt, GSK-3β, p-GSK-3β, and p-Akt. ResultCompared with the normal group, the modeling elevated the serum levels of AST, CK, cTnT, and MYO (P<0.01), destroyed the arrangement of myocardial cells abd nucle, twisted and broken myocardial fibers, up-regulated the protein levels of LC3Ⅱ/Ⅰ and Beclin-1 (P<0.01), and down-regulated the protein levels of PI3K, p-Akt, and p-GSK-3β (P<0.01). Compared with the model group, trimetazidine and Wutou Chishizhi Wan (all the doses) lowered the levels of AST, CK, cTnT, and MYO in serum (P<0.01), restored the arrangement of myocardial cells and muscle fibers, reduced necrosis, down-regulated the protein level of Beclin-1 (P<0.01), and up-regulated the protein levels of PI3K, p-Akt, and p-GSK-3β (P<0.01). Additionally, Wutou Chishizhi Wan (all the doses) down-regulated the protein level of LC3Ⅱ/Ⅰ (P<0.05, P<0.01). Compared with those in the trimetazidine group, the serum AST level rose in the low-dose Wutou Chishizhi Wan group (P<0.05) and declined in the high-dose group (P<0.01), and the protein level of Beclin-1 was down-regulated in the medium-dose group (P<0.01). Additionally, the trimetazidine group had higher protein level of LC3Ⅱ/Ⅰ than medium- and high-dose Wutou Chishizhi Wan groups (P<0.05, P<0.01), higher protein level of PI3K than low-, medium-, and high-dose groups (P<0.01), lower protein level of p-Akt than low- and medium-dose groups (P<0.01), and higher p-GSK-3β protein level than the medium-dose group (P<0.01). ConclusionDifferent doses of Wutou Chishizhi Wan can ameliorate MIRI, and the high dose has the best effect. Wutou Chishizhi Wan can reduce the activity of myocardial injury markers AST, CK, cTnT, and MYO, and alleviate the pathological damage of myocardial tissue. It can down-regulate the protein levels of beclin-1, LC3Ⅱ/Ⅰ, and up-regulate those of PI3K, p-Akt, and p-GSK-3β. In summary, Wutou Chishizhi Wan may inhibit excessive autophagy and regulate the PI3K/Akt/GSK-3β signaling pathway to exert protective effect on MIRI rats.
5.The necessity of post-maneuver postural restriction in treating posterior canal benign paroxysmal positional vertigo: a Meta-analytic study.
Jing Jing WANG ; Yong TANG ; Cheng YU ; Xi Long GUAN ; Yong Sheng LIU ; Yi Wen XUE
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2021;56(6):603-612
Objective: We conducted a Meta-analysis to investigate the necessity of postural restrictions after manual reduction in the treatment of posterior canal benign paroxysmal positional vertigo (PC-BPPV). Methods: We searched PubMed, EBSCO, Proquest, Web of Science databases, Ovid, and screened eligible studies that investigated the effect of post-maneuver postural restriction in treating patients with PC-BPPV. Outcomes included the efficacy of treatment and recurrence. Meta-analysis was performed using Stata 15.0 software. Results: Studies of the single visit efficiency included 11 references, with a sample size of 1 733 cases. The Meta-analysis results showed that the difference in the efficacy between the postural restricted group and the non-postural restricted group in PC-BPPV patients was statistically significant(RR=1.12, 95%CI=1.07-1.18, P<0.001). There were 12 references included in the study on the total efficiency, with a cumulative sample size of 1763 cases. There was no statistically significant difference between the effect of postural restriction after manipulative reduction and that of simple manipulative reduction (RR=1.03, 95%CI=0.99-1.08, P=0.118). There were 5 references included in the study of recurrence rate, and the cumulative sample size was 659 cases. There was no statistically significant difference in the recurrence rate between the postural restricted group and the non-postural restricted group(RR=0.98, 95%CI=0.62-1.54, P=0.937). Conclusions: In comparison with non-postural restriction group, post-maneuver postural restriction after a single visit can improve the treatment effective rate of PC-BPPV and contribute to the improvement in the symptoms of patients in a short term. However, postural restrictions has no significant effect on the final prognosis of PC-BPPV, and it also has no significant effect on the recurrence.
Benign Paroxysmal Positional Vertigo/therapy*
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Environment
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Humans
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Patient Positioning
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Recurrence
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Semicircular Canals
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Treatment Outcome
6.Biomass carbohydrates assimilation and lipid accumulation by Mortierella isabellina.
Dahui XING ; Anlong PAN ; Donghua XUE ; Mingyu FANG ; Ranfeng GU
Chinese Journal of Biotechnology 2010;26(2):189-193
Biomass carbohydrates assimilation and lipid accumulation by Mortierella isabellina M2 strain were investigated. Corn fiber hydrolysate was specially studied. The results showed M. isabellina M2 strain achieved growth and lipid accumulation while glucose, xylose, mannose and arabinose were introduced as single carbon source, respectively. When M. isabellina M2 strain was cultivated on corn fiber hydrolysate with 6% sugars concentration, the biomass reached 18.2 g/L, the lipid content of dry mycelia was 45.7%, and the lipid yield achieved 8.3 g/L. It provided a promising perspective for microbial oils production with biomass hydrolysates.
Biomass
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Carbohydrate Metabolism
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Carbon
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metabolism
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Fermentation
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Industrial Microbiology
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methods
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Lipids
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biosynthesis
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Mortierella
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metabolism
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Zea mays
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metabolism
7.Effect of culture conditions on cell growth and lipid accumulation of oleaginous microorganism.
Lijuan MA ; Dahui XING ; Honglei WANG ; Xiaojun WANG ; Donghua XUE
Chinese Journal of Biotechnology 2009;25(1):55-59
We used four strains (Y1, Y2, M1 and M2) to screen out the high lipid production strains. We first adopted cell morphology and cytochemical methods (Sudan III dyeing technique) to observe intracellular characteristics. Observation results indicated that M2 strain had the potential lipid accumulation capacity. To prove this, lipid content of these strains was determined by soxhlet extraction. One strain (M2) was found to produce lipids up to 53.09%. In order to increase the production of oleaginous microorganism, the effects of hydrolysate concentration, nitrogen source, pH, fermentation temperature and time on cell growth and lipid accumulation were studied. The optimal fermentation conditions were obtained as follows: corn starch byproduct hydrolysate concentration at 10(o) Bx as carbon source; NaNO3 as nitrogen source at 0.2%; initial pH of 6.0; temperature at 28 degrees C, cultivated for 6 d. Under these conditions, M2 strain accumulated lipids up to 75.21% on a cellular biomass basis with biomass yield of 30.40 g/L, and the corresponding lipid production reached 22.86 g/L. GC analysis demonstrated that the fatty acid composition of the lipid was similar to that of vegetable oil, which mainly contained 16-and 18-carbon fatty acids. Thus, microbial lipid is a promising material for biodiesel production, and its unsaturated fatty acid content reached around 68%. These unsaturated fatty acids show great potential applications in food, medicine and cosmetics industries.
Biomass
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Culture Media
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Fatty Acids
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analysis
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biosynthesis
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Fermentation
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Hydrogen-Ion Concentration
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Industrial Microbiology
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Starch
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metabolism
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Temperature
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Yeasts
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growth & development
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metabolism
8.Study on Synthesis and Identification of Rotundic Acid Esters Derivatives and in vitro Antitumor Activity
Minlun NAN ; Yufang HE ; Xueling SI ; Yuwei ZHAO ; Xue WANG ; Xue BAI ; Chuanjing LI
China Pharmacy 2019;30(5):591-595
OBJECTIVE: To synthesize and identify rotundic acid esters derivatives, and to evaluate in vitro antitumor activity. METHODS: Using rotundic acid as raw material, rotundic acid esters derivatives were synthesized by 28-esterification, 3β and 23-position hydroxyl group combined with acid anhydride. The structure of rotundic acid esters derivatives were identified by HRMS, 1H-NMR, 13C-NMR and physicochemical properties. The in vitro antitumor activities were evaluated by MTT assay against paclitaxel (positive control), rotundic acid and rotundic acid esters derivatives with human cervical cancer cells HeLa, human malignant melanoma cells A375, human lung adenocarcinoma cells SPC-A1 and human liver cancer cells HepG2 (take IC50 as an indicator). RESULTS: Totally 6 kinds of rotundic acid esters derivatives were synthesized, i.e. rotundic acid methyl ester(compound 1), 3, 23-O-di(acetyl) rotundic acid methyl ester(compound 2), 3, 23-O-di(propionyl) rotundic acid methyl ester(compound 3), 3, 23-O-di(butyryl) rotundic acid methyl ester(compound 4), 3, 23-O-di(phthaloyl) rotundic acid methyl ester(compound 5) and 3, 23-O-di(succinyl) rotundic acid methyl ester(compound 6). Compared with paclitaxel, the antitumor activity of compound 5 and compound 6 to Hela, A375, HepG2 and SPC-A1 was similar to it, and there was no significant difference between them (P>0.05); the IC50 of rotuntic acid and other compounds decreased with statistical significance (P<0.05). CONCLUSIONS: In the study, 6 kinds of rotundic acid esters derivatives are synthesized and compound 5 and compound 6 have significant antitumor activity in vitro, which can provide reference for further study.
9.Application of Westgard sigma rule in quality control laboratory of blood station
Shanshan LIU ; Xiaotang YU ; Xue JIANG ; Di CHI
Chinese Journal of Blood Transfusion 2022;35(1):93-94
【Objective】 To help quality control laboratory of blood station select suitable internal quality control rules using Westgard sigma rules. 【Methods】 The accumulated coefficient of variation of internal quality control was used as the measurement imprecision in quality control laboratory of blood station, and the bias of the results of EQA in 2020 was regarded as Bias. The allowable total error (TEa) of WS/T406-2012 was used as the evaluation index to calculate the σvalue of laboratory blood testing items to select reasonable and feasible quality control rules using the Westgard sigma rule. 【Results】 The average σvalues of total protein in the three national EQAS were 14.2, 8.7 and 9.6, respectively. The average σvalues of fibrinogen in two national EQAS were 3.6 and 4.1. The average σvalues of Plt counts and MCHC were <4 and >3, and those of other items were more than 6 in two national EQAS. 【Conclusion】 The rule of 1
10. Synthesis of Echinocystic Acid Derivatives and Inhibitory Effect on Lipase
Xue WANG ; Min-lun NAN ; Xue BAI ; Yu-fang HE ; Yu-wei ZHAO ; Chuan-jing LI ; Zhong-mei HE
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(20):125-130
Objective:Echinocystic acid(EA)is a kind of oleanolic pentacyclic triterpenoid compound,due to its main structural features of stability and less active sites,the structures of EA were modified in this paper to synthesize a series of EA derivatives, improve their bioavailability, and investigate their inhibitory effect on lipase. Method:In this study,EA derivatives were designed and synthesized from EA,which is a natural lipase inhibitor. Their inhibitory effects on lipase were tested by using 2,4-dinitrophenyl butanoate(PNPB) method. Result:Nine compounds were synthesized,and their structures were characterized by infrared spectrum (IR), ultraviolet spectrum (UV), mass spectrum (MS), nuclear magnetic resonance spectrum (1H-NMR and 13 C-NMR),all of which were identified as new compounds. Further experiments on the inhibitory effect on lipase showed that compounds 1-9 had higher inhibitory effects than EA,IC50=7.03,2.05,2.14,3.65,3.24,0.28,0.34,0.46,and 0.39 g·L-1. Compounds 6-9 had higher inhibitory effect than Orlistat(IC50=0.53 g·L-1). Inhibition rates were as follows:6 > 7 > 9 > 8 > Orlistat> 2 > 3 > 5 > 4 > 1 >EA. Conclusion:It is feasible to design and synthesize derivatives with EA as the lead compound to improve the inhibitory effect on lipase.