1.Protective effect of gastrodin on A β 25-35-induced HT22 cell injury by improving glycolytic function through activating PI3K/Akt/BACH1 signaling axis
Tingting LIU ; Xinhao JI ; Aihong WEI ; Huiping LEI ; Lingnan WU ; Shaoyu ZHOU ; Jingshan SHI ; Feng JIN
Chinese Journal of Pharmacology and Toxicology 2023;37(7):505-505
OBJECTIVE To investigate whether gas-trodin(GAS)plays a neuroprotective role by activating PI3K/Akt/BACH1 signaling axis to improve glycolytic func-tion.METHODS HT22 cells were treated with Aβ25-35 for 24 h to establish cell damage model.GAS pretreated HT22 cells for 2 h,and Akt agonist SC79,Akt inhibitor MK2206,PI3K inhibitor LY294002 were added 0.5 h before GAS treatment to detect their protective mecha-nisms.Pharmacodynamic research of GAS in this model were divided into six groups:control group,GAS group(GAS 10 μmol·L-1),model group(Aβ25-35 20 μmol·L-1),model +GAS 2.5,5 and 10 μ mol·L-1 group).Mecha-nism research of GAS in this model was divided into 6 groups:control group,Aβ25-35 20 μmol·L-1 group,Aβ25-35 20 μmol·L-1 + GAS 10 μmol·L-1 group,Aβ25-35 + SC79 group(Aβ25-35 20 μmol·L-1 +SC79 10 μmol·L-1),Aβ25-35+MK2206+GAS group(A β 25-35 20 μ mol·L-1 +MK2206 10 μmol·L-1+GAS 10 μmol·L-1),Aβ25-35+LY294002+GAS group(Aβ25-35 20 μmol·L-1+LY294002 10 μmol·L-1+GAS 10 μmol·L-1).Cell viability was detected by MTT,mor-phological changes of cells were observed by micro-scope,ATP content was detected by chemilumines-cence,and pyruvate(PA)content was detected by colo-rimetry.Western blotting was used to detect the protein levels of transcription factor BACH1,key glycolysis enzyme hexokinase(HK1)and PI3K/Akt signaling path-way related proteins PI3K,p-PI3K,Akt and p-Akt.RESULTS The results showed that compared with the control group,the cell morphology of HT22 cells damaged by Aβ25-35 was damaged,the number of cells decreased,the cell body became smaller,the number of dead cells increased,the cell survival rate,ATP and PA contents decreased significantly,and the protein expressions of p-PI3K,p-Akt,BACH1 and HK1 were significantly down-regulated.GAS treatmentcansignificantlyimprovethemor-phology of HT22 cells damaged by Aβ25-35,increase cell survival rate,ATP and PA contents,and up-regulate the expression of p-PI3K,p-Akt,BACH1 and HK1 proteins.SC79 also significantly increased cell survival rate,ATP content,protein expression of BACH1 and HK1.However,the above ameliorative effect of GAS on HT22 cell dam-age induced by Aβ25-35 was antagonized by LY294002 and MK2206.CONCLUSION GAS exerts a neuroprotec-tive effect on Aβ25-35-induced HT22 cell injury by improv-ing glycolytic function through activating PI3K/Akt/BACH1 signaling axis.
2.Analysis of the salt-stress responsive element of the promoter of peanut small GTP binding protein gene AhRabG3f.
Guoning DU ; Jie XIANG ; Shunyu LIN ; Xiangyuan KONG ; Xiuling WU ; Xuedong GUAN ; Hong ZHU ; Jingshan WANG ; Lixian QIAO ; Jiongming SUI ; Chunmei ZHAO
Chinese Journal of Biotechnology 2022;38(8):2989-2998
To study the molecular mechanism of salt stress response of peanut small GTP binding protein gene AhRabG3f, a 1 914 bp promoter fragment upstream of the start codon of AhRabG3f gene (3f-P) from peanut was cloned. Subsequently, five truncated fragments (3f-P1-3f-P5) with lengths of 1 729, 1 379, 666, 510 and 179 bp were obtained through deletion at the 5' end, respectively. Plant expression vectors where these six promoter fragments were fused with the gus gene were constructed and transformed into tobacco by Agrobacterium-mediated method, respectively. GUS expression in transgenic tobacco and activity analysis were conducted. The gus gene expression can be detected in the transgenic tobacco harboring each promoter segment, among which the driving activity of the full-length promoter 3f-P was the weakest, while the driving activity of the promoter segment 3f-P3 was the strongest. Upon exposure of the transgenic tobacco to salt stress, the GUS activity driven by 3f-P, 3f-P1, 3f-P2 and 3f-P3 was 3.3, 1.2, 1.9 and 1.2 times compared to that of the transgenic plants without salt treatment. This suggests that the AhRabG3f promoter was salt-inducible and there might be positive regulatory elements between 3f-P and 3f-P3 in response to salt stress. The results of GUS activity driven by promoter fragments after salt treatment showed that elements included MYB and GT1 between 1 930 bp and 1 745 bp. Moreover, a TC-rich repeat between 682 bp and 526 bp might be positive cis-elements responsible for salt stress, and an MYC element between 1 395 bp and 682 bp might be a negative cis-element responsible for salt stress. This study may facilitate using the induced promoter to regulate the salt resistance of peanut.
Arachis/genetics*
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Fabaceae/genetics*
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GTP-Binding Proteins/metabolism*
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Gene Expression Regulation, Plant
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Glucuronidase/metabolism*
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Plant Proteins/metabolism*
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Plants, Genetically Modified/genetics*
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Salt Stress
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Stress, Physiological/genetics*
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Tobacco/genetics*
4.Value of soluble triggering receptor expression on myeloid cells-1 level of alveolar fluid in early diagnosis of ventilator-associated pneumonia: a Meta-analysis
Jiayu TAN ; Xiang HUANG ; Minzhi WU ; Bo YANG ; Zhengchao LUO ; Jingshan LI ; Yuanfang ZHONG ; Zhenpeng LIANG
Chinese Critical Care Medicine 2020;32(9):1067-1073
Objective:Serum soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) is a useful biomarker of bacterial infection. However, the diagnostic value of sTREM-1 of alveolar fluid in pulmonary infection is still unclear. This article aimed to explore the value of sTREM-1 of alveolar fluid in the early diagnosis of ventilator-associated pneumonia (VAP) by systematic review of relevant literatures.Methods:CNKI, Wanfang, VIP, PubMed/Medline and Embase databases were retrieved. Articles on diagnosis of VAP by sTREM-1 before June 30, 2019 were collected. QUADAS-2 scale provided by Cochrane Collaboration Network was used to evaluate the quality of diagnostic experiments. RevMan 5.3 and Stata 13.0 software were used to complete Meta-analysis. The levels of sTREM-1 between VAP and non-VAP patients were analyzed by Meta-analysis, and then diagnostic test Meta-analysis was conducted. Heterogeneity, sensitivity and publication bias were analyzed.Results:A total of 24 articles were enrolled. QUADAS-2 scale indicated that the selected literature had low bias and high clinical adaptability. ① In Meta-analysis of sTREM-1 level in alveolar fluid, 20 articles were selected and found to have high heterogeneity ( I2 = 94.4%, P = 0.000). The random effects models were used for Meta-analysis. It was indicated that the sTREM-1 level in alveolar fluid of VAP group was significantly higher than that of non-VAP group with significant difference [standardized mean difference ( SMD) was 1.47, 95% confidence interval (95% CI) was 1.00-1.95, Z = 6.14, P = 0.000]. By subgroup analysis and Meta-regression analysis, no source of heterogeneity was found. Sensitivity analysis suggested that the results of this Meta-analysis were robust and credible, and Begg funnel plot analysis showed that there was no significant publication bias ( Z = 1.46, P = 0.143). ② A total of 18 articles were included in the Meta-analysis of diagnostic experiments. Deek funnel plot showed publication bias ( P = 0.012). The combined sensitivity was 0.87 (95% CI was 0.81-0.91), specificity was 0.80 (95% CI was 0.73-0.86), and diagnostic odds ratio ( DOR) was 26 (95% CI was 13-50). Subgroup analysis of three different sources of alveolar fluid (bronchoalveolar lavage fluid, endotracheal aspiration fluid and exhaled ventilator condensate) showed that STREM-1 had a certain value in early diagnosis of VAP. The I2 of combined DOR was 35.4%, and I2 of sensitivity was 79.46%, I2 of specificity was 77.61%, suggesting heterogeneity in the selected literature. Subgroup analysis found that nationality, subject design, sample source, sample size and diagnostic "gold criteria" were related to heterogeneity, but not age. The area under synthetic receiver operating characteristic (SROC) curve (AUC) was 0.90 (95% CI was 0.87-0.92). Conclusions:The detection of sTREM-1 level in alveolar fluid can be used for the early diagnosis of VAP with high sensitivity and specificity. If combined with other biomarkers, it may have more diagnostic value.
5.Breeding on a new peanut variety Yuhua91 with high oleic acid content.
Leilei PAN ; Yanan JIANG ; Wenjie ZHOU ; Pingping JIANG ; Lanrong WU ; Ao CHEN ; Hong ZHU ; Jiongming SUI ; Jingshan WANG ; Lixian QIAO
Chinese Journal of Biotechnology 2019;35(9):1698-1706
Yuhua91 is a new peanut variety with high oleic acid content bred by Qingdao Agricultural University. The crossing was conducted with Luhua11 as female parent and with Kainong1715, an F435-type variety with high oleic acid content as male parent. The real F1 hybrids were screened by sequencing on PCR amplification products, and those homozygotes with bb genotype in F2 populations were screened by the same sequencing method as above. The content of oleic and linoleic acid was measured on the kernels harvested from F2 single plants by near infrared ray method, and those kernels whose content of oleic was above 80%, oleic and linoleic acid ratio was above 10.0 were obtained and planted into a row, with pedigree method for subsequent selection breeding. Yuhua91 has some characters of small pod, light and obvious pod texture, 148.06 g per 100 pods, 63.31 g per 100 kernels, 75.15% shelling percentage, long elliptic seed kernel, pink seed coat, without crack, white endotesta. Its content of protein, oil, oleic acid, linoleic acid and palmitic acid was 26.57%, 52.72%, 80.40%, 2.50% and 5.57% respectively. Yuhua91 has other characters of strong seedlings, compact pod areas, and moderate resistance to leaf spot disease and bacterial wilt. Average pod yield is 215.79 kg per Mu, 15.27% higher than the control variety Huayu20. Average seed kernels yield is 157.33 kg per Mu, 21.64% higher than the control variety Huayu20. Yuhua 91 has been registered on department of agriculture in 2018, and the registration No. is GPD peanut (2018) 370210, fit for growing in Shandong Province.
Arachis
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Oleic Acid
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Plant Breeding
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Seeds
6.Effect of early intervention of vacuum sealing drainage under laparoscopy on inflammatory mediators in patients with severe acute pancreatitis
Zhihao ZHUANG ; Shiming TAO ; Jingshan HUO ; Minhan WU ; Yanchuan LU ; Rizhao WU ; Xinfeng HAN
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2018;25(2):157-160
Objective To investigate the effect of section of pancreatic envelope combined with vacuum sealing drainage under laparoscopy on inflammatory mediators of patients with early severe acute pancreatitis (SAP). Methods Forty-two SAP patients were admitted to Foshan Hospital of Traditional Chinese Medicine in Guangdong Province from January 2008 to December 2016. That 22 patients underwent pancreatic membrane incision and vacuum sealing drainage under laparoscopy was in the experimental group, and that 20 patients underwent the routine pancreatic membrane incision and double tube drainage was in the control group. The venous blood was collected, the levels of C-reactive protein (CRP), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured before and after operation for 1, 3, 7 and 14 days, and the clinical therapeutic effects were observed in the two groups. Results With the prolongation of therapy, the levels of CRP, IL-6 and TNF-α were decreased continuously in both groups, the degrees of decrease of above indexes in experimental group were more significant than those in the control group, and the differences in these indexes between the two groups were statistically significant [CRP (mg/L): 1 day was 203.80±25.12 vs. 271.79±60.41, 3 day was 117.26±19.70 vs. 174.53±42.37; IL-6 (ng/L): 1 day was 40.63±3.38 vs. 57.10±11.46, 3 days was 23.14±3.51 vs. 46.87±10.69; TNF-α (ng/L): 1 day was 23.91±10.42 vs. 36.73±15.90, 3 days was 19.13±8.34 vs. 32.58±15.81, all P < 0.05]. There were no statistical significant differences in the levels of above indexes on 7 days and 14 days after treatment between the two groups (all P > 0.05). The therapeutic efficacy of the experimental group was significantly higher than that of the control group [95.45% (21/22) vs. 90.0% (18/20), P < 0.05]. Conclusion Under laparoscopy, pancreatic envelope incision combined with vacuum sealing drainage performed for early SAP patients can control the body inflammation more rapidly, reduce complications and shorten the disease course.
7.Meta-analysis of Shenling Baizhu Powder Combined with Aminosalicylic Acid Preparation in the Treatment of Ulcerative Colitis
Kerui WU ; Jingshan LUO ; Jiangfeng WU ; Tianming HE ; Lixin TAN ; Xiantao LI
China Pharmacy 2017;28(36):5119-5122
OBJECTIVE:To evaluate clinical efficacy of Shenling baizhu powder combined with aminosalicylic acid in the treatment of ulcerative colitis (UC),and to provide evidence-based reference in clinic.METHODS:Retrieved from PubMed,Cochrane library,CJFD,VIP,Wanfang database and CBM,randomized controlled trials (RCTs) about Shenling baizhu powder combined with aminosalicylic acid preparations (trial group) vs.aminosalicylic acid preparations alone (control group) in the treatment of UC were collected.Meta-analysis was performed by using Rev Man 5.2 statistical software afder data extraction and quality evaluation with Cochrane systematic evaluator manual 5.1.0.RESULTS:A total of 14 RCTs were included,involving 1 177 patients.Meta-analysis showed that total response rate of trial group was significantly higher than that of control group [OR=3.40,95% CI (2.44,4.74),P<0.001],while Sutherland DAI score [MD=-1.31,95% CI (-2.07,-0.56),P<0.001],the levels of TNF-α [SMD=-1.90,95%CI(-2.90,-0.90),P<0.001],IL-17[MD=-85.18,95%CI(-100.51,-69.85),P<0.001] and IL-23 [MD=-113.27,95%CI(-132.34,-94.21),P<0.001] in trial groups were significantly lower than control group,with statistical significance.CONCLUSIONS:Therapeutic efficacy of Shenling baizhu powder combined with aminosalicylic acid preparation is better than aminosalicylic acid preparation alone in the treatment of UC,and can significantly reduce Suthedand DAI and the levels of serum cytokines.
8.The preliminary research of T2 mapping at 3.0T MR in lumbar intervertebral disc degeneration
Jingshan TAO ; Xianhua WU ; Feng HAN ; Juan ZHANG ; Yingdi XIE
Journal of Practical Radiology 2016;(2):288-291
Objective The purpose of our research was to quantitative study the degeneration of lumbar intervertebral discs by using T2 mapping technology at 3.0T MR and research the correlation of biomechanics and lumbar disc degeneration.Methods 34 patients with low back pain or sciatica were enrolled.The correlation of T2 values of lumbar intervertebral discs and Pfirrmann grading,disc location,patient ages were analyzed.T2 values before and after loading were also analyzed.Results T2 values of nucleus pulposus were correlation with Pfirrmann scoring,disc location and,patient ages.Conclusion (1 )T2 mapping provides a non-invasive and quantitative way to diagnose lumbar discs degeneration.(2)The change of external pressure load can affect the T2 values of the intervertebral disc.
9.Protective effects of NDP on LPS induced neuron injuries in rat mixed cultures
Mu LIN ; Qihai GONG ; Qin WU ; Feng ZHANG ; Jingshan SHI
Chinese Pharmacological Bulletin 2016;32(8):1144-1147,1148
Aim To investigate the protective effect of noble dendrobium polysaccharides ( NDP ) on lipopo-lysaccharide ( LPS)-induced neuron injuries in newborn rat cerebral cortex glial cells and neuron mixed cul-tures.Methods The primary cultures of newborn rat cortical neurons and glial cells were established and the existence of the neurons , astrocytes and microglia was verified respectively .NDP was given to LPS-induced mixed cultures , the mRNA levels of IL-1β, TNF-αand COX-2 were assayed by real time PCR .Results NDP reduced the glial cell activation and neuron dam-age after it was given to LPS-induced mixed cultures . The mRNA levels of IL-1β, TNF-α, COX-2 were re-duced .Conclusion NDP protects against LPS-in-duced neuron-inflammation in neurons and glial cells cultures.
10.Analysis of Piper puberulum (Benth.) Maxim.extracts preparation and the preliminary qualitative chemical composition
Yan YANG ; Qin WU ; Qihai GONG ; Shibing XU ; Jingshan SHI
Chinese Journal of Biochemical Pharmaceutics 2015;37(5):169-173
Objective To investigate optimal extraction process of Piper puberulum ( Benth.) Maxim.and qualitative analyze the chemical component of the extracts.Methods Method of solvent heating reflux was used for extraction.On the basis of single factor experiment, L9 (34 ) orthogonal experiment was designed with the variants of extraction frequency, time, material-liquid ratio, and immersion time.Extraction rate as index, extraction processes were optimized to achieve best extraction.The extracts, including total extract, water elution, and ethanol elution, were physiochemically analysed to achieve an initial qualitative result.Results The optimal extraction process was: extractions 3 times for 2 hours, with an 1︰30 material -liquid ratio and 2 hours of immersion, Initial qualitative analyzed the total extracts containing amino acids, polypeptides, proteins, alkaloids, steroids or triterpenes, flavones, saponins, polysaccharides, reducing sugars or glucosides, cumarins, terpene lactones, phenols, and tannins.The water elution containing: amino acids, polypeptides, proteins, saponins, polysaccharides, reducing sugars or glucosides, cumarins, and terpene lactones.The ethanol elution containing: amino acids, polypeptides, proteins, alkaloids, steroids or triterpenes, flavones, polysaccharides, reducing sugars or glucosides, phenols, and tanins.Conclusion The experiments show that optimal extraction process can achieve high extraction yield, stable and practical.

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