1.Protective Mechanism of Paeoniflorin on Mice with Ulcerative Colitis Based on AMPK/mTOR Autophagy Pathway
Xin DAI ; Rou LI ; Yang HU ; Yuhang WANG ; Ruizhu ZHAO ; Jiaxuan FENG ; Shilei LOU ; Cong SUN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(3):45-53
ObjectiveTo explore the protective mechanism of paeoniflorin on mice with ulcerative colitis (UC) through the adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) autophagy pathway. MethodUC mouse model was established by allowing mice freely drink 4% DSS, and 56 BALB/c male mice were randomly divided into model group, AMPK inhibitor group (20 mg·kg-1), paeoniflorin (50 mg·kg-1) + inhibitor (20 mg·kg-1) group, and high dose (50 mg·kg-1), medium dose (25 mg·kg-1), and low dose (12.5 mg·kg-1) paeoniflorin groups. After seven days of drug intervention, the protective effect of paeoniflorin on mice with UC was determined by comparing the body weight, disease activity index (DAI) changes, and Hematoxylin-eosin (HE) staining results. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the serum of mice in each group, and immunofluorescence was utilized to detect microtubule-associated protein 1 light chain 3 (LC3) content in the colon, AMPK, mTOR proteins, and their phosphorylated proteins including p-AMPK and p-mTOR in the colon tissue were detected by Western blot, and the mRNA expression levels of AMPK, mTOR, Beclin1, LC3, and p62 were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultCompared with the blank group, the model group showed a decrease in body mass, an increase in DAI score, and severe pathological damage to the colon. The levels of inflammatory factors including TNF-α and IL-6 increased in serum (P<0.01), while the protein levels of LC3 and p-AMPK/AMPK were down-regulated in colon tissue, and those of p-mTOR/mTOR were up-regulated (P<0.01). The mRNA expression levels of AMPK and LC3 were down-regulated, while the mRNA expression levels of mTOR and p62 were up-regulated (P<0.01). Compared with the model group and the paeoniflorin + inhibitor group, the mice treated with paeoniflorin showed an increase in body mass, a decrease in DAI score, a reduction in pathological damage to colon tissue, and a reduction in the levels of inflammatory factors of TNF-α and IL-6 in serum (P<0.05). The protein levels of LC3 and p-AMPK/AMPK in colon tissue were up-regulated, while the protein levels of p-mTOR/mTOR were down-regulated (P<0.01). The mRNA expression levels of AMPK, Beclin1, and LC3 were up-regulated, while the mRNA expression of mTOR and p62 were down-regulated (P<0.01). The colon tissue of the inhibitor group was severely damaged, and the trend of various indicators was completely opposite to that of the high dose paeoniflorin group. ConclusionPaeoniflorin can enhance autophagy and reduce inflammatory damage in mice with UC by activating the AMPK/mTOR signaling pathway and thus play a protective role.
2.Mechanism of Huanglian Jiedutang in Inhibiting Activation of NLRP3 Inflammasomes and Ameliorating Acute Liver Injury in Septic Mice
Ruizhu ZHAO ; Zhengyang HUA ; Yuhang WANG ; Xinyue REN ; Dingxing FAN ; Shilei LOU ; Hui YAN ; Cong SUN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(22):27-34
ObjectiveTo explore the mechanism of Huanglian Jiedutang in inhibiting the pyroptosis mediated by NOD-like receptor protein 3 (NLRP3) inflammasomes and alleviating the acute liver injury (ALI) induced by lipopolysaccharide (LPS) in the mouse model of sepsis. MethodFifty-four male C57BL/6 mice were randomized into blank, model, low- (3.08 g·kg-1), medium- (6.15 g·kg-1), and high-dose (12.30 g·kg-1) Huanglian Jiedutang, and positive control (dexamethasone) groups (n=9). The mice were administrated with Huanglian Jiedutang at different doses by gavage for 7 days, and then LPS (15 mg·kg-1) was injected intraperitoneally for the modeling of sepsis. In the positive control group, dexamethasone (0.05 g·kg-1) was injected intraperitoneally 1.5 h after modeling, and the mouse sepsis score (MSS) was recorded 12 h after modeling. The mice were sacrificed for the collection of blood and liver tissue samples. The levels of alanine transaminase (ALT) and aspartate transaminase (AST) were measured by a biochemical analyzer. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β, and IL-18 in the serum were measured by enzyme-linked immunosorbent assay kits. Hematoxylin-eosin staining was used to observe the pathological changes in the liver tissue. The content of NLRP3 was observed by the immunofluorescence assay. The expression of apoptosis-associated speck-like protein containing CARD (ASC) was detected by immunohistochemistry. The protein levels of NLRP3, ASC, Caspase-1, and gasdermin D (GSDMD) in the liver tissue were determined by Western blot. Real-time quantitative polymerase chain reaction(Real-time PCR) was employed to determine the mRNA levels of GSDMD, Caspase-1, IL-1β, and IL-18. ResultCompared with the blank group, the model group showed elevated levels of ALT and AST (P<0.01) and risen levels of inflammatory cytokines in the serum (P<0.01). In addition, the modeling resulted in edema and necrosis in the liver, and up-regulated the protein levels of GSDMD, NLRP3, ASC, and Caspase-1 (P<0.01) and the mRNA levels of GSDMD, Caspase-1, IL-1β, and IL-18 (P<0.01). Compared with the model group, the drug intervention groups showed reduced content of inflammatory cytokines (P<0.01), alleviated pathological damage in the liver tissue, and down-regulated protein levels of GSDMD, NLRP3, ASC, and Caspase-1 (P<0.05,P<0.01) and mRNA levels of GSDMD, Caspase-1, IL-1β, and IL-18 (P<0.05,P<0.01) in the liver tissue. ConclusionHuanglian Jiedutang can inhibit pyroptosis and reduce inflammation by inhibiting the activation of NLRP3 inflammasomes, thus demonstrating a therapeutic effect on acute liver injury in the mouse model of sepsis induced by LPS.
3.The evaluation value of ultrasound measurement of optic disc combined with optic nerve sheath in patients with severe high intracranial pressure
Zeyang YU ; Xiaonan SONG ; Cong LI
Journal of Apoplexy and Nervous Diseases 2021;38(11):976-979
To explore whether ultrasound measurement of optic disc height can evaluate intracranial hypertension and the value of ultrasound measurement of optic nerve sheath diameter combined with optic disc height in the diagnosis of elevated intracranial hypertension. Methods Enrolled patients who needed lumbar puncture in the Department of Neurology of the First Hospital of Jilin University between July 2019 and November 2020.Complete ultrasound examination before lumbar puncture,measure ONSD and ODH. According to the lumbar puncture pressure,the patients were divided into normal intracranial pressure group and intracranial hypertension group. Singlefactor and multifactor analysis were performed to obtain the relationship between ICP and each ultrasound data. Analyze the correlation between ICP and ONSD and ODH. Generate ROC curve,evaluate and compare the value of ONSD,ODH and ONSD combined with ODH in diagnosing intracranial hypertension. Results A total of 78 patients,39 cases in each group. There was no statistical difference in baseline data between the two groups. (1) ONSD[(5.04±0.34) mm]in intracranial hypertension group is higher than normal intracranial pressure group[(4.17±0.38) mm](P<0.001);ODH[0.81 (0.59-1) mm]in intracranial hypertension group is higher than normal intracranial pressure group[0.39 (0-0.59) mm](P<0.001);(2)Correlation analysis showed that ICP was positively correlated with ONSD (r=0.779,P<0.001)and ODH(r=0.572,P<0.001);(3) The cutoff value of ODH for diagnosing elevated ICP is 0.54 mm,the sensitivity and specificity are 85% and 72%,respectively;the cutoff value of ONSD for diagnosing elevated ICP is 4.68 mm,the sensitivity and specificity are 87% and 95%,respectively;The sensitivity and specificity of ONSD combined with ODH in the diagnosis of elevated ICP are 95% and 95%,respectively;(4)Compare the value of the three in diagnosing elevated ICP,ONSD is greater than ODH (P=0.036),ONSD combined with ODH is greater than ONSD(P=0.043)or ODH (P=0.001) alone. Conclusion Ultrasound measurement of ODH can be used to diagnose intracranial hypertension.ONSD combined with ODH can improve the sensitivity of diagnosing elevated ICP.
4.A case of type 2 diabetes with glycogenic hepatopathy
Renshu MA ; Tong WU ; Ge YU ; Ying CONG ; Guijie XIN
Journal of Clinical Hepatology 2021;37(9):2183-2184
6.Clinicopathological features and prognostic analysis of 151 patients with primary extra-nodal non-Hodgkin's lymphoma.
Cong WANG ; Changji YUAN ; Hua HE ; Ou BAI
Chinese Journal of Oncology 2014;36(11):858-862
OBJECTIVETo evaluate the epidermiology, clinicopathological features and prognosis of primary extra-nodal non-Hodgkin's lymphoma (PE-NHL).
METHODSThe clinicopathological data of 151 patients diagnosed as PE-NHL in the First Affiliated Hospital of Jilin University between January 2007 and December 2011 were reviewed and analyzed.
RESULTSThe proportion of PE-NHL patients was 58.8% among all the non-Hodgkin's lymphoma cases, with the average age of 52 years, and the male/female ratio was 1.16:1. The most frequently involved site was the stomach (30.5%) followed by nose and sinuses (19.9%) and Waldeyer's ring (17.2%). The most common subtype was diffuse large B-cell lymphoma (DLBCL) (55.0%), followed by NK/T (18.5%) and extra-nodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) ( 13.2%). The distribution of clinical stages was as follows: stage I 20.5%, II 29.8%, III 29.1%, and IV 20.5%. Most nasal PE-NHL is in early stages, with a proportion of 76.7% in stages I & II. The 3-year overall survival rate was 73.2% and 3-year progression free survival rate was 46.6% in the PE-DLBCL patients. The International Prognosis Index (IPI) could be used to estimate the prognosis of PE-DLBCL well. Multivariate analysis showed that ESR and curative effect of the first treatment were independent prognostic factors for PE-DLBCL patients.
CONCLUSIONSThe incidence of PE-NHL is quite high, and the most common primary extra-nodal site is the gastro-intestinal tract and the most common subtype is diffuse large B-cell lymphoma. Risk groups based on IPI can indicate the prognosis of PE-DLBCL to some extent, but only the ESR and curative effect of the first treatment are confirmed to be independent risk factors.
Adult ; Disease-Free Survival ; Female ; Humans ; Lymphoma, B-Cell ; Lymphoma, B-Cell, Marginal Zone ; Lymphoma, Large B-Cell, Diffuse ; Lymphoma, Non-Hodgkin ; diagnosis ; pathology ; Male ; Multivariate Analysis ; Prognosis ; Retrospective Studies ; Survival Rate
7.Studies on antigencity of human immunodeficiency virus type 1 (HIV-1) external glycoprotein as well as its expression in Pichia pastoris.
Li-Hui ZHAO ; Xiang-Hui YU ; Chun-Lai JIANG ; Yong-Ge WU ; Jia-Cong SHEN ; Wei KONG
Chinese Journal of Biotechnology 2007;23(3):457-461
Based on the computer simulation, we analyzed hydrophobicity, potential epitope of recombined subtypes HIV-1 Env protein (851 amino acids) from Guangxi in China. Compared with conservative peptides of other subtypes in env protein, three sequences (469-511aa, 538-674aa, 700-734aa) were selected to recombine into a chimeric gene that codes three conservative epitope peptides with stronger antigencity, and was constructed in the yeast expression plasmid pPICZB. Chimeric proteins were expressed in Pichia pastoris under the induction of methanol, and were analyzed by SDS-PAGE and Westernblot. The results showed that fusion proteins of three-segment antigen were expressed in Pichia pastoris and that specific protein band at the site of 40kD was target protein, which is interacted with HIV-1 serum. The target proteins were purified by metal Ni-sepharose 4B, and were demonstrated to possess good antigenic specificity from the data of ELISA. This chimeric antigen may be used as research and developed into HIV diagnostic reagents.
Blotting, Western
;
Enzyme-Linked Immunosorbent Assay
;
Gene Expression Regulation, Viral
;
HIV Seronegativity
;
immunology
;
HIV Seropositivity
;
immunology
;
HIV-1
;
genetics
;
immunology
;
metabolism
;
Humans
;
Pichia
;
genetics
;
Recombinant Fusion Proteins
;
genetics
;
immunology
;
metabolism
;
env Gene Products, Human Immunodeficiency Virus
;
genetics
;
immunology
;
metabolism
8.The effect of basic fibroblast growth factor on the gene expression of decorin by periodontal ligament fibroblasts in culture.
Si-cong WANG ; Chong-tao LIN ; Dai-bang NIE ; Hong-sheng OUYANG
West China Journal of Stomatology 2008;26(4):352-354
OBJECTIVETo study the effect of basic fibroblast growth factor (bFGF) on the gene expression of decorin by periodontal ligament fibroblasts (PLFs) in culture, and discuss the effect of bFGF in periodontal regeneration.
METHODSHuman PLFs were cultured and stimulated by exogenous bFGF. Gene expression of decorin was assessed by semi-quantitive RT-PCR.
RESULTSThe mRNA expression of decorin was suppressed by bFGF and the effect was dose-dependent. When the dose of bFGF increased, the inhibitive effect decreased.
CONCLUSIONDecorin has many biological effects. The inhibitive effect may be one of important factors which participate in the healing process of periodontitis, and provide partly theoretical basis of bFGF in periodontal regeneration.
Decorin ; Fibroblast Growth Factor 2 ; Fibroblasts ; Humans ; Periodontal Ligament ; RNA, Messenger ; Regeneration
9.Early use of calf pulmonary surfactant in late preterm and full-term infants with respiratory distress syndrome: a randomized controlled trial.
Wen-Li ZHOU ; Qi ZHOU ; Cong LI ; Hui WU
Chinese Journal of Contemporary Pediatrics 2014;16(3):285-289
OBJECTIVETo evaluate the efficacy of calf pulmonary surfactant (PS) in the treatment of respiratory distress syndrome (RDS) in late preterm and full-term infants.
METHODSA randomized controlled trial was designed to evaluate the efficacy of calf PS intratracheally given at different times and doses in infants with RDS who had a gestational age of ≥35 weeks and an oxygenation index (OI) of 10-20. The subjects were randomly assigned to treatment group 1 (n=58), treatment group 2 (n=58), and control group (n=59). Treatment group 1 was given PS (50 mg/kg) within 6 hours after admission. Treatment group 2 was given PS (70 mg/kg) within 6 hours after admission. The control group was not given PS within 6 hours after admission and was given PS (50 mg/kg) over 6 hours after admission if having no remission by conventional therapy including mechanical ventilation. For each group, a second dose of PS (50 mg/kg) was given if no remission was observed within 12 hours after the first administration.
RESULTSThere were no significant differences in mortality between the three groups. Treatment group 2 had lower hospitalization expense and shorter duration of mechanical ventilation compared with treatment group 1, and treatment group 1 had lower hospitalization expense and shorter duration of mechanical ventilation compared with the control group. The incidence of ventilator-associated pneumonia and length of hospital stay in treatment group 2 was lower than those in treatment group 1 and control group. Compared with the control group, Treatment groups 1 and 2 showed decreases in 2 or more times of PS use, maximum OI, duration of continuous positive airway pressure treatment, and incidence of air leak syndrome and pulmonary hypertension.
CONCLUSIONSEarly use of sufficient PS in late preterm and full-term infants with RDS can reduce complications, secondary use of PS, duration of mechanical ventilation and length of hospital stay, and hospitalization expense.
Female ; Hospitalization ; economics ; Humans ; Infant, Newborn ; Infant, Premature ; Male ; Pulmonary Surfactants ; therapeutic use ; Respiratory Distress Syndrome, Newborn ; drug therapy
10.Expression of SOX4 gene and early recurrence of hepatocellular carcinoma: their relationship and the clinical significance.
Jiang-hua ZHENG ; Zhi-xiang JIAN ; Hao-sheng JIN ; Si-cong CHEN ; Guan-yi WANG
Journal of Southern Medical University 2010;30(4):818-819
OBJECTIVET To explore the relationship between the expression of SOX4 gene and early recurrence of hepatocellular carcinoma (HCC) after curative resection.
METHODSSOX4 expression was detected immunohistochemically in 60 HCC patients including 30 with and 30 without early recurrence after curative resection, with 30 normal liver specimens as the control.
RESULTSThe expression of SOX4 was significantly higher in HCC than in normal liver (41.7% vs 16.7%, P<0.05), and in HCC tissues, the expression was significantly higher in early recurrent HCC after curative resection than in HCC without early recurrence (56.7% vs 26.7%, P<0.05). SOX4 expression was inversely correlated to the patients' gender, age, tumor size, HBsAg, and Edmonson grade (P<0.05).
CONCLUSIONSOX4 is closely associated with early recurrence of HCC after curative resection, and its overexpression may contribute to early recurrence of HCC. SOX4 may serve as a new molecular indicator for evaluating the prognosis of HCC.
Adult ; Aged ; Biomarkers, Tumor ; Carcinoma, Hepatocellular ; genetics ; metabolism ; surgery ; Female ; Humans ; Liver Neoplasms ; genetics ; metabolism ; surgery ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; genetics ; Prognosis ; SOXC Transcription Factors ; genetics ; metabolism