1.Inhibitory effects of special siRNA targeting TLR4 gene on the TNF-α expression of BV-2 cells induced by hypoxia-reoxygenation
Peng SUN ; Jiyuan HAN ; Qing ZHANG ; Jinghui ZHANG ; Yuan TIAN
Chinese Journal of Emergency Medicine 2009;18(3):270-273
Objective To study the inhibition of tumor necrosis factor-alpha(TNF-α)cytokine expression of BV-2 cells induced by hypoxia-reoxygenation injury by siRNA targeting TLR4 gene via the RNAi mechanisms.Method BV-2 mouse microglial cell line was cultured in six-well plates and randomly divided into group N(nor-mal group),group H(hypoxia-reoxygenation),group T(hypoxia-reoxygenation+TLR4-siRNA transfected group),group C(hypoxia-reoxygenation+pEGFP-H1/control-siRNA transfected group)and group B(hypox-ia-reoxygenation+pEGFP-H1 transfected group).Group H,group T,group C and group B were cultured in hy-poxia condition for 3 h followed by reoxygenation for 24 h.The plasma was transfected into BV-2 cells mediated by lipofectamine 2000.The efficiency of transfection were detected by flow cytometry to observe the expression of EGFP.RT-PCR method was used to detect the level of mRNA of TLR4 or NF-кB p65.Westem blot methed was used to test the expression of TLR4 protein.and ELISA was used to test the level of TNF-α in the supernatants.Analysis of variances was used for statistical analysis.Results The expression of EGFP gene waa;(67.58±7.16)% after transfection by flow cytometry analysis.Compared to group N,the TLR4 mRNA,NF-кB p65 mR-NA,TLR4 protein level and the TNF-α quantity in group H,group T,group C and group B increased after the hy-poxia-reoxygenation treatment(P<0.01).While the expression of the TLR4 mRNA,NF-кB p65 mRNA,TLR4 protein level and the TNF-α quantity in the group T down-regulated compared to group H,group C and group B(P<0.01).And there were no changes in group C,group B and group H about observation index(P>0.05).Conclusions The siRNA targeting TLR4 mRNA could inhibit the inflammatory reaction released by BV-2 cells in-duced by hypoxia-reoxygenation stimulation.
2.Effects of ggpS over-expression on glycosylglycerol and glycerol biosynthesis of Synechocystis sp. PCC 6803.
Peizhen MA ; Xiaoming TAN ; Xuefeng LÜ ; Jiyuan TIAN
Chinese Journal of Biotechnology 2016;32(3):347-354
To study the roles of glucosylglycerol phosphate synthase (Ggps) in glucosylglycerol (GG) and glycerol biosynthesis, we over-expressed Ggps from either Synechocystis sp. PCC 6803 or Synechococcus sp. PCC 7002 in a Synechocystis strain with a high GG titer, and determined the GG and glycerol accumulation in the resultant mutants grown under different NaCl-stress conditions. Ion chromatography results revealed that GG yield was not improved, but glycerol production was significantly enhanced by over-expression of Ggps from Synechocystis sp. PCC 6803 (6803ggpS). In addition, increasing the NaCl concentration of medium from 600 to 900 mmol/L led to a further 75% increase of glycerol accumulation in the mutant strain with 6803ggpS over-expression. These findings show the role of ggpS in driving the carbon flux to the glycerol biosynthesis pathway, and will be helpful for further improvement of GG and glycerol production in Synechocystis.
Bacterial Proteins
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metabolism
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Culture Media
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Glucosides
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biosynthesis
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Glucosyltransferases
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metabolism
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Glycerol
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metabolism
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Industrial Microbiology
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Sodium Chloride
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Synechococcus
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enzymology
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Synechocystis
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enzymology
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metabolism
3.Expression of somatostatin receptor subtypes and epidermal growth factor receptor in non-small cell lung cancer and their clinical significance
Xin LIU ; Huifang TIAN ; Xiuwen WANG ; Jiyuan DING
Chinese Journal of Cancer Biotherapy 1994;0(01):-
0.05).There was a negative correlation between the EGFR expression with the expression of SSTR2A and SSTR5 in NSCLC tissues.The 3-year survival rates were 64.5% and 65.9% in patients positive of SSTR5 and SSTR2A,respectively;and were 45.2% and 22.2% for those negative of SSTR5 and SSTR2A,respectively(P
4.Clinical Observation of Fire-needle Acupuncture for Sub-acute and Chronic Eczema
Yang CHENG ; Xiaoyong ZHOU ; Xianyu ZENG ; Jiyuan WU ; Wen LIU ; Liming TIAN ; Kai LI ; Qingjun TIAN ; Shijie BAO
Shanghai Journal of Acupuncture and Moxibustion 2014;(10):903-905
Objective To compare the therapeutic efficacies of fire-needle acupuncture and Halometasone cream in treating sub-acute and chronic eczema. Method Totally 114 patients were divided into a treatment group and a control group. The treatment group was intervened by using fire-needle acupuncture, and the control group was by Halometasone cream. Itchy score and Eczema Area and Severity Index (EASI) were used for comparison before and after intervention. Result There were significant differences in comparing the markedly-effective rate and total effective rate between the fire-needle acupuncture group and the Halometasone control group according to the itchy score (P<0.01). There were no significant differences in comparing the markedly-effective rate and total effective rate between the two groups according to EASI (P>0.05). Conclusion Fire-needle acupuncture is superior to Halometasone cream for acute and chronic eczema in relieving itch, action time and effect-sustaining duration.
5.Expression and their clinical significance of SSTR2A, SSTR5 and EGFR in non-small cell lung cancer.
Xin LIU ; Huifang TIAN ; Xiuwen WANG ; Jiyuan DING
Chinese Journal of Lung Cancer 2007;10(1):14-19
BACKGROUNDSomatostatin receptor (SSTR), as a marker gene in tumors, is being valued gradually. With five subtypes have been identified, many researches are carried out to explore the amino acid sequence of SSTR family members, the molecular biological characteristics, the distribution and expression in the normal tissue and the tumor, and their specific ligands. In this study, the expression and significance of SSTR (SSTR2A, SSTR5) and epidermal growth factor receptor (EGFR) in human non-small cell lung cancer (NSCLC) were investigated, and the relationship among them were evaluated.
METHODSThe expressions of SSTR2A, SSTR5 and EGFR in 62 NSCLC tissues and 7 lung tissues adjacent to the cancer tissues were detected by immunohistochemical method (SP method). All cases were followed up.
RESULTSIn 62 cases of NSCLC, the positive rate of SSTR2A and SSTR5 expression was 48.4% (30/62) and 71.0% (44/62) respectively. The positive rate of SSTR2A and SSTR5 was closely related to TNM stage (P < 0.05), but not to other clinical characteristics of NSCLC (P > 0.05). The positive rate of EGFR expression was 56.5% (35/62), but 0 in 7 lung tissues adjacent to the cancer tissues. The positive rate of EGFR was not related to the age, sex, smoking or not, tumors histological type, tumor size, TNM stage, differentiation classification and the lymph node metastasis (P > 0.05). There was negative relation between the expression of SSTR2A, SSTR5 and EGFR in NSCLC. The 3-year survival rate of patients with SSTR2A and SSTR5 expression was 64.52% and 65.91% respectively, 45.16% and 22.22% for those without expression (P < 0.05); The 3-year survival rate of patients with EGFR expression was 30.77% and 69.44% for those without expression (P < 0.05 ).
CONCLUSIONSThe expression of SSTR and EGFR is significantly upregulated in NSCLC and a negative relation exists between their expressions. Detection of expression of SSTR2A, SSTR5 and EGFR might be helpful to evaluate lymph node metastasis, pathological stages and prognosis of NSCLC.
6.Predictive value of lactate concentration combined with lactate clearance rate in the prognosis of neonatal septic shock
Yi WANG ; Jiahao TIAN ; Xuefeng YANG ; Sixiu LI ; Jiyuan GUO
Chinese Journal of Pediatrics 2021;59(6):489-494
Objective:To investigate the predictive value of lactate concentration within 1 h after admission combined with lactate clearance rate (LC) at 6 h after fluid resuscitation in prognosis of neonatal septic shock.Methods:In this retrospective study, 58 newborns with septic shock admitted to the Neonatal Intensive Care Unit of Xi′an Children′s Hospital,Xi′an Jiao Tong University from June 2016 to March 2020 were enrolled. According to the mortality within 60 days after admission,which was also set as the end point, the patients were divided into death group and survival group. The general demographic data and clinical variables including blood cell counts, procalcitonin, C-reactive protein, D-dimer, serum creatinine, and lactate concentration within 1 h after admission (Lac1) and at 6 h after fluid resuscitation (Lac2) were collected. The differences in the clinical variables between the survival and death group were compared by independent sample t test or Rank-Sum test, and the risk factors of poor prognosis were analyzed by binary Logistic regression. The predictive values of these risk factors were tested by receiver operating characteristic (ROC) curve. Furthermore, the cut-off of the risk factors were used to analyze the accumulative survival rate by Kaplan-Meier curve. Results:A total of 58 neonates were enrolled, among whom 24 survived and 34 died within 60 days after admission. The rate of premature rupture of membranes in the death group was higher than that in the survival group (41% (14/34) vs.13%(3/24), P=0.021). There were also significant differences in infection site, pathogenic characteristics, total fluid volume of resuscitation, vasoactive drug index, rate and complications of mechanical ventilation between the two groups (all P<0.05). The levels of Lac1, Lac2, procalcitonin, D-dimer and serum creatinine in the death group were higher than those in the survival group ((12±6) vs. (7±4) mmol/L, (14±6) vs. (4±2) mmol/L, (59±23) vs.(24±14) ng/L, (24±11) vs.(11±6) mg/L, (167±31) vs.(92±23) μmol/L, t=3.549, 3.112, 3.859, 4.499, 3.288, all P<0.05). While the blood pressure and LC at 6 h after fluid resuscitation were lower than those in the survival group ((41±12) vs. (52±5) mmHg (1 mmHg=0.133 kPa), t =4.230;-16 (-40, 20) % vs. 40 (18, 70) %, Z= 3.558, all P<0.05). Binary Logistic regression analysis showed that LC was negatively associated with the risk of death in neonates with septic shock (odds ratio ( OR) and 95% confidence interval ( CI): 0.679 (0.662-0.999), P<0.05), while Lac1 was the risk factor and positively associated with the risk of death ( OR and 95% CI: 1.203 (0.965-1.500), P<0.05). Furthermore, the predictive values of LC, Lac1 and the combination of these two variables in the prognosis of neonatal septic shock were analyzed by ROC curve analysis, and the area under the curve (AUC) were 0.699, 0.875, 0.965, respectively, with the sensitivity of 83.32%, 89.65% and 94.31%, and the specificity of 72.52%, 77.18% and 88.76%, respectively. According to the cut-off value of Lac1, the newborns with Lac1>4 mmol/L had significantly lower accumulative survival rate than those with Lac1≤4 mmol/L by Kaplan-Meier analysis (21% (8/38) vs. 80% (16/20), χ 2=54.520, P<0.05). According to the cut-off value of LC, the newborns with LC ≤ 10% had significantly lower accumulative survival rate than those with LC>10% by Kaplan-Meier analysis (19% (6/32) vs. 69% (18/26), χ 2=14.140, P<0.05). Conclusion:The combination of lactate concentration and lactate clearance rate have an optimal predictive value in the prognosis of neonatal septic shock.
7.Intervention Effect of Suanzaoren Tang on Depression Model Rats Based on JNK/c-Myc/p53 Pathway
Shuailin DU ; Zhicheng HAO ; Ce ZHANG ; Jiyuan GUO ; Xusheng TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):12-19
ObjectiveTo investigate the intervention effects of Suanzaoren Tang on depression model rats induced by isolation combined with chronic unpredictable mild stress (CUMS), and to examine its influence on the c-Jun N-terminal kinase (JNK)/proto-oncogene protein (c-Myc)/tumor suppressor protein 53 (p53) signaling pathway, thereby revealing its potential functional mechanism. MethodsA total of 72 male SD rats were randomly divided into six groups using a strict random number table: blank group, model group, fluoxetine group (3.6 mg·kg-1), and high-, medium-, and low-dose Suanzaoren Tang groups (10, 5, 2.5 g·kg-1),with 12 rats in each group. A depression model was established using isolation combined with CUMS. Fluoxetine and different doses of Suanzaoren Tang were administered continuously for 28 days. Behavioral indicators such as sucrose water consumption and open field test scores were recorded. Western blot and immunohistochemistry (IHC) were employed to analyze the expression of key proteins in the JNK/c-Myc/p53 signaling pathway, and the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was used to evaluate the number of apoptotic cells in the hippocampus. ResultsCompared with the blank group, the model group exhibited a significantly reduced sucrose preference index (P<0.01), a lower total score of horizontal and vertical movements in the open field test (P<0.01), significantly increased expression of JNK, c-Myc, and p53 proteins in the hippocampus (P<0.01), and a higher number of TUNEL-positive cells in the hippocampus (P<0.01). Compared with the model group, the sucrose preference index and the total score of horizontal and vertical movements in the open field test significantly increased in the high- and medium-dose Suanzaoren Tang groups and the fluoxetine group (P<0.05, P<0.01). The expression of JNK, c-Myc, and p53 proteins significantly decreased in all Suanzaoren Tang groups (high, medium, and low doses) and the fluoxetine group (P<0.05, P<0.01). The number of TUNEL-positive cells in the hippocampus also significantly decreased in these groups (P<0.01). ConclusionSuanzaoren Tang can regulate the expression of JNK/c-Myc/p53 proteins in the hippocampus of depression model rats, and its antidepressant mechanism may be related to its protective effect on hippocampal neurons.
8.Development and validation of a nomogram for predicting survival of breast cancer patients with ipsilateral supraclavicular lymph node metastasis.
Min-Hao LYU ; You-Zhao MA ; Pei-Qi TIAN ; Hui-Hui GUO ; Chao WANG ; Zhen-Zhen LIU ; Xiu-Chun CHEN
Chinese Medical Journal 2021;134(22):2692-2699
BACKGROUND:
Breast cancer patients with ipsilateral supraclavicular lymph node metastasis (ISLNM) but without distant metastasis are considered to have a poor prognosis. This study aimed to develop a nomogram to predict the overall survival (OS) of breast cancer patients with ISLNM but without distant metastasis.
METHODS:
Medical records of breast cancer patients who received surgical treatment at the Affiliated Cancer Hospital of Zhengzhou University, Jiyuan People's Hospital and Huaxian People's Hospital between December 21, 2012 and June 30, 2020 were reviewed retrospectively. Overall, 345 patients with pathologically confirmed ISLNM and without evidence of distant metastasis were identified. They were further randomized 2:1 and divided into training (n = 231) and validation (n = 114) cohorts. A nomogram to predict the probability of OS was constructed based on clinicopathologic variables identified by the univariable and multivariable analyses. The predictive accuracy and discriminative ability were measured by calibration plots, concordance index (C-index), and risk group stratification.
RESULTS:
Univariable analysis showed that estrogen receptor-positive (ER+), progesterone receptor-positive (PR+), human epidermal growth factor receptor 2-positive (HER2+) with Herceptin treatment, and a low axillary lymph node ratio (ALNR) were prognostic factors for better OS. PR+, HER2+ with Herceptin treatment, and a low ALNR remained independent prognostic factors for better OS on multivariable analysis. These variables were incorporated into a nomogram to predict the 1-, 3-, and 5-year OS of breast cancer patients with ISLNM. The C-indexes of the nomogram were 0.737 (95% confidence interval [CI]: 0.660-0.813) and 0.759 (95% CI: 0.636-0.881) for the training and the validation cohorts, respectively. The calibration plots presented excellent agreement between the nomogram prediction and actual observation for 3 and 5 years, but not 1 year, OS in both the cohorts. The nomogram was also able to stratify patients into different risk groups.
CONCLUSIONS
In this study, we established and validated a novel nomogram for predicting survival of patients with ISLNM. This nomogram may, to some extent, allow clinicians to more accurately estimate prognosis and to make personalized therapeutic decisions for individual patients with ISLNM.
Breast Neoplasms
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Female
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Humans
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Lymph Nodes
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Lymphatic Metastasis
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Nomograms
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Retrospective Studies