1.Effect of South African Herb Hoodia gordonii on Liver Glucolipid Metabolism and PI3K/Akt/FoxO1 Signalling Pathway in db/db Mice
Chengfei ZHANG ; Qiue ZHANG ; Linging QIN ; Wei LIU ; Guangyuan XU ; Xuesheng MA ; Lili WU ; Tonghua LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):57-64
ObjectiveTo observe the effects of the South African herb Hoodia gordonii (HG) on glucolipid metabolism in diabetic db/db mice and explore the possible mechanisms of HG on the liver of db/db mice based on the phosphoinositide-3 kinase (PI3K)/protein kinase B (Akt)/factor forkhead protein O1 (FoxO1) signaling pathway. MethodA total of 30 db/db mice were randomly divided into five groups according to fasting blood glucose: model group, metformin group (0.195 g·kg-1), and low dose (0.39 g·kg-1), medium dose (0.78 g·kg-1), and high dose (1.56 g·kg-1) HG groups, with six m/m mice in each group, and another six m/m mice were set as normal group. The mice in the normal and model groups were given saline of 9 mL·kg-1 by gavage. Body weight, water intake, and fasting blood glucose of the mice in each group were measured weekly. After six weeks of continuous administration, serum insulin (FINS), low-density lipoprotein cholesterol (LDL), total cholesterol (TC), triglyceride (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea, and creatinine (CREA) were measured, and liver sections were embedded and stained with hematoxylin-eosin (HE), periodic acid-Schiff (PAS), and oil red O. Protein expression of PI3K p85, p-Akt, and p-FoxO1 in liver was detected by immunohistochemistry. The mRNA expression of PI3K, Akt, and FoxO1 in liver tissue was detected by real-time polymerase chain reaction (Real-time PCR). ResultAfter six weeks of administration intervention, it was found that fasting blood glucose was significantly downregulated in mice in the three HG groups (P<0.05). The level of islet resistance index was significantly reduced in both the low and medium dose HG groups (P<0.05). The expression levels of TC, TG, and LDL were reduced in all HG groups (P<0.05, P<0.01). Pathologically, HG could alleviate hepatocyte steatosis, reduce the volume and content of lipid droplets in liver, and increase the distribution of glycogen granules in liver to some extent in mice. Immunohistochemical assays revealed that PI3K p85 protein expression was significantly increased in the low, medium, and high dose HG groups compared with the model group (P<0.01). p-Akt protein expression was significantly increased in the medium and high dose HG groups (P<0.05, P<0.01). p-FoxO1 protein expression was significantly increased in the low, medium, and high dose HG groups (P<0.05, P<0.01). Compared with the model group, PI3K mRNA was increased in low dose, medium dose, and high dose HG groups (P<0.05), and Akt mRNA was increased in high dose HG group (P<0.05). FoxO1 mRNA was decreased in low dose, medium dose, and high dose HG groups (P<0.05). ConclusionHG can ameliorate the disorder of glucolipid metabolism in db/db mice, which may be related to its activation of the hepatic PI3K/Akt/FoxO1 signaling pathway.
2.Mechanism of Flavonoids from Mulberry Leaves on Insulin Resistance and Inflammatory Response in Diabetic Mice
Guangyuan XU ; Xiaoming ZHANG ; Chunling JIA
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(16):52-57
ObjectiveTo observe the glucose-lowering, insulin resistance-improving, and anti-inflammatory effects of flavonoids from mulberry leaves (FML) and explore their underlying mechanism. MethodMale db/db mice aged 6-7 weeks were randomly divided into a model group, a high-dose FML group (1.00 g·kg·d-1), and a low-dose FML group (0.50 g·kg-1·d-1). C57BL mice of the same age were assigned to the normal group. After six weeks of intervention, fasting blood glucose (FBG), serum fasting insulin levels (Fins), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), free fatty acid (FFA), blood creatinine (SCr), blood urea nitrogen (BUN), and aspartate aminotransferase (AST) levels were measured, and the homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase activities in the liver were measured. Morphological changes in the liver were assessed by hematoxylin-eosin (HE) staining. The protein expression of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and nuclear factor-κB (NF-κB) in the liver was detected by Western blot. ResultCompared with the model group, the high-dose and low-dose FML groups showed significant reductions in FBG, Fins, HOMA-IR, IL-6, TNF-α, and FFA levels (P<0.05, P<0.01), and increased levels of SOD, GSH-Px, and catalase in the liver (P<0.05, P<0.01). HE staining of the liver in the FML groups showed improved arrangement of hepatocytes, reduced inflammatory cell infiltration, and alleviated cellular steatosis compared with the model group. The protein expression of COX-2, iNOS, and NF-κB in the liver significantly decreased in the FML groups as compared with that in the model group (P<0.05, P<0.01). ConclusionFML have glucose-lowering and insulin resistance-improving effect, which may be attributed to their regulation of the NF-κB pathway in the liver of diabetic mice, leading to the suppression of the release of COX-2, iNOS, and inflammatory cytokines, thereby improving the inflammatory state.
3.Mechanism of Momordica charantia Extract in Regulating Gluconeogenesis in ZDF Diabetes Rats
Guangyuan XU ; Zhuo ZHANG ; Xiaoming ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(16):65-71
ObjectiveTo observe the effect of Momordica charantia extract (MCE) on the gluconeogenesis signaling pathway in diabetes rats. MethodMale Zucker Diabetic Fatty (ZDF) rats aged 5-6 weeks were randomly divided into a model group and an MCE group (administered MCE at a dose of 0.40 g·kg-1 by gavage). Additionally, seven healthy male ZDF (fa/+) rats were assigned to the normal group and received administration once daily for six consecutive weeks. During the experiment, the general condition of the rats was observed, and body weight was recorded. Fasting blood glucose and random blood glucose levels were measured in the 1st, 3rd, and 5th weeks. In the 6th week, an oral glucose tolerance test (OGTT) was conducted, and serum levels of triglycerides (TG), free fatty acid (FFA), total cholesterol (TC), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were measured. Hematoxylin-eosin (HE) staining was performed to examine liver morphology, periodic acid-Schiff (PAS) staining was used to assess hepatic glycogen storage, and Real-time polymerase chain reaction (PCR) was employed to measure the mRNA expression of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) in the liver. Western blot analysis was conducted to measure the phosphorylation level of forkhead box protein O1 (FoxO1) and the protein expression of PEPCK and G6Pase in the liver. ResultCompared with the model group, the MCE group showed significant improvements in body weight, fasting blood glucose, random blood glucose, and glucose tolerance (P<0.05, P<0.01) and reduced serum levels of FFA, TC, and TG (P<0.05, P<0.01). There were no significant differences in ALT and AST between the two groups. In the MCE group, the HE staining revealed more orderly liver cell arrangement and reduced hepatic steatosis and the PAS staining showed increased hepatic glycogen storage. The protein expression of p-FoxO1 in the liver was significantly elevated (P<0.01), while there was no significant difference in FoxO1 protein expression. The mRNA and protein expression of PEPCK and G6Pase significantly decreased (P<0.05). ConclusionMCE exhibits glucose-lowering and lipid-lowering effects, improves glucose tolerance, and enhances hepatic glycogen storage. These effects may be attributed to the upregulation of p-FoxO1, leading to the inhibition of PEPCK and G6Pase expression and the regulation of gluconeogenesis-related processes.
4.Effects of chitinase-3-like protein 1 on the expression of inflammatory damage-related molecules in mouse skeletal muscle satellite cells induced by lipopolysaccharide
Fuxing LI ; Zhiyong SHENG ; Yihua ZHOU ; Jingying LI ; Jianning XU
Chinese Critical Care Medicine 2021;33(12):1428-1433
Objective:To explore the potential mechanism of chitinase-3-like protein 1 (CHI3L1) involved in skeletal muscle stem cell injury induced by sepsis.Methods:Six different concentrations of lipopolysaccharide (LPS) were used to stimulate mouse skeletal muscle satellite cells cultured in vitro. Enzyme linked immunosorbent assay (ELISA) and cell counting kit-8 (CCK-8) were used to determine the optimal concentration. The overexpression and interference vectors of CHI3L1 were constructed to transfect skeletal muscle satellite cells, and the transfection efficiency was verified by polymerase chain reaction (PCR) and Western blotting. The cells were randomly divided into blank control group (cells without any intervention), model group (LPS-stimulated untransfected cells), overexpressing CHI3L1 group (LPS-stimulated cells transfected with CHI3L1 plasmid), overexpressing CHI3L1 control group [LPS-stimulated cells transfected with negative control (NC) plasmid], CHI3L1 interference group [LPS-stimulated cells transfected with CHI3L1 small interfering RNA (siRNA)], CHI3L1 interference control group (LPS-stimulated cells transfected with CHI3L1-siRNA NC). The levels of extracellular caspase-1 and interleukin-1β (IL-1β) were detected by ELISA. The protein expressions of intracellular IL-1β, signal transducer and activator of transcription 3 (STAT3), protein kinase B (Akt) and phosphorylated Akt (p-Akt) were detected by Western blotting. Results:According to the results of CCK-8 and ELISA, the best concentration of 5 mg/L LPS was selected for the subsequent experiment. The transfection was validated by PCR and Western blotting. Compared with the blank control group, the levels of extracellular IL-1β, caspase-1 and the protein expressions of intracellular Akt, p-Akt, and IL-1β were significantly increased in the model group [IL-1β (ng/L): 11.22±0.55 vs. 8.63±0.63, caspase-1 (pmol/L): 9.47±0.22 vs. 8.65±0.15, Akt/GAPDH: 1.36±0.12 vs. 1.06±0.15, p-Akt/GAPDH: 0.78±0.07 vs. 0.09±0.01, IL-1β/GAPDH: 1.38±0.12 vs. 0.18±0.03, all P < 0.05]. Compared with the model group and the overexpressing CHI3L1 control group, the levels of extracellular IL-1β, caspase-1 and the protein expressions of intracellular p-Akt and IL-1β were significantly increased in the overexpressing CHI3L1 group [IL-1β(ng/L): 14.93±0.97 vs. 11.22±0.55, 9.38±0.40, caspase-1 (pmol/L): 10.35±0.03 vs. 9.47±0.22, 8.46±0.24, p-Akt/GAPDH: 1.21±0.04 vs. 0.78±0.07, 0.63±0.04, IL-1β/GAPDH: 1.87±0.08 vs. 1.38±0.12, 1.51±0.17, all P < 0.05]. Compared with the model group and the CHI3L1 interference control group, the levels of extracellular IL-1β, caspase-1 and the protein expressions of intracellular p-Akt and IL-1β were significantly decreased in the CHI3L1 interference group [IL-1β(ng/L): 8.98±0.73 vs. 11.22±0.55, 10.44±0.65, caspase-1 (pmol/L): 7.61±0.63 vs. 9.47±0.22, 8.37±0.38, p-Akt/GAPDH: 0.50±0.04 vs. 0.78±0.07, 0.94±0.06, IL-1β/GAPDH: 0.77±0.02 vs. 1.38±0.12, 1.13±0.07, all P < 0.05]. Conclusions:CHI3L1 may mediate the damage of skeletal muscle stem cells in sepsis by increasing the expression of caspase-1 and IL-1β. CHI3L1 may be involved in the regulation of Akt signaling pathway in skeletal muscle stem cells, but has no significant effect on STAT3 signaling pathway.
5. Clinical Efficacy of Yinchenhao Tang Combined with Sinisan in Treating Nonalcoholic Fatty Liver Disease with Hyperhomocysteinemia
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(16):61-66
Objective: To investigate the clinical efficacy of Yinchenhao Tang combined with Sinisan(YCSN) on non-alcoholic fatty liver disease (NAFLD) with hyperhomocysteinemia (HHcy). Method: Totally 126 cases of NAFLD with HHcy in Beijing Shichahai Community Health Service Center were randomly divided into NAFLD control group, traditional Chinese medicine(TCM) group, and modern medicine group, with 42 cases in each group. The TCM group was treated with YCSN orally, the modern medicine group was treated with polyene phosphatidylcholine capsules and folic acid tablets, and the NAFLD control group was treated with placebo. The course of treatment was 3 months. The clinical efficiency, liver function, Hcy, blood lipid, oxidative inflammation index and symptom score in two groups were observed before and after treatment. Result: The total effective rate of the TCM group was 92.86%, while that of the modern medicine group was 88.10%. There was no significant difference in the clinical efficiency between the two groups. There was no difference in the indexes of groups before treatment. After treatment,alanine aminotransferase (ALT), aspartate aminotransferase (AST), Hcy, total cholesterol (TC), total triglyceride (TG), low-density lipoprotein (LDL), malondialdehyde (MDA), high-sensitive C reactive protein (hs-CRP), serum creatinine (SCr), blood urea nitrogen (BUN), and symptom scores in the TCM group and the modern medicine group were significantly lower than those before the treatment (P<0.05), and lower than those in the NAFLD control group (P<0.05); and high-density lipoprotein (HDL) and superoxidase dismutase (SOD) in the TCM group and the modern medicine group significantly increased (P<0.05), and higher than those of NAFLD control group (P<0.05). Compared with the modern medicine group, ALT, Hcy, TC, TG, MDA, hs-CRP, and symptom scores in TCM group were significantly lower than those before the treatment (P<0.05), while SOD in the TCM group was significantly increased (P<0.05). Conclusion: YCSN can improve hyperhomocysteinemia, liver function, blood lipid, oxidation and inflammation in patients with NAFLD and HHcy.
6.Risk factors of pathological discrepancy between biopsy and excisional specimen from gastric low-grade intraepithelial neoplasia and early gastric cancer
Feng LI ; Ping XIANG ; Qi OUYANG ; Fuxing XU ; Renxiang HUANG ; Zili XIAO ; Danian JI ; Yun ZHOU ; Tao SUN
Chinese Journal of Digestive Endoscopy 2018;35(5):336-340
Objective To investigate the risk factors of pathological discrepancy between biopsy and excisional specimen from gastric low-grade intraepithelial neoplasia (LGIN) and early gastric cancer (EGC). Methods A retrospective analysis was conducted on the data of 235 patients who underwent endoscopic submucosal dissection or surgical resection and diagnosed as LGIN or EGC ( including high-grade intraepithelial neoplasia) by postoperative pathology. Patients were grouped by whether there was significant pathological discrepancy between biopsy and excisional specimen. Univariate and multivariate analyses were used to analyze the risk factors for significant pathological discrepancy. Results Significant pathological discrepancy occurred in 33 cases (14. 0%). Univariate analysis showed that protruding lesion, non-reddish surface, without erosion or ulcer, diffused pathological type and number of biopsy were related to the pathological discrepancy (all P<0. 05). Multivariate analysis suggested that small number of biopsy blocks (OR=0. 574, 95%CI: 0. 363-0. 908, P=0. 018) was an independent risk factor for significant pathological discrepancy. Conclusion The pathological discrepancy between biopsy and excisional specimen from gastric LGIN and EGC are common. Multiple biopsies can improve the accuracy of biopsy and reduce the occurrence of pathological discrepancy with excisional specimen.
7.Performance Investigation of Ion Trap with Various Collision gas and Pressures
Fuxing XU ; Qiankun DANG ; Hangyu DING ; Zhengxu HUANG ; Yuanyuan WANG ; Zhen ZHOU ; Chuanfan DING
Chinese Journal of Analytical Chemistry 2017;45(4):587-592
Ion trap performances are investigated based on digital ion trap technique with different collision gases at different pressures. Collision gases of helium (4 amu), nitrogen (28 amu) and argon (40 amu) with various pressures are adopted in ion excitation and dissociation stages to investigate the ion trap performances, including mass resolution, signal intensity, tandem mass spectrometric analysis ability and low-mass cut off (LMCO) effect. It is found that when heavy gas of argon is used, energy can be efficiently transferred and LMCO effect is decreased with higher ion capture and dissociation efficiencies but with low mass resolution. Higher mass resolution is realized with helium as collision gas. Furthermore, at the same gas pressure, heavy gas is beneficial to abundant fragment ions and structural information of precursor ion.
8.The effects of multiple intravenous tranexamic acid administrations after total knee arthroplasty on fibrinolytic activity and inflammatory response
Shaoyun ZHANG ; Jinwei XIE ; Qiang HUANG ; Bin XU ; Guorui CAO ; Fuxing PEI
Chinese Journal of Orthopaedics 2017;37(23):1483-1489
Objective To explore the effects of multiple intravenous tranexamic acid (Ⅳ-TXA) administrations after total knee arthroplasty (TKA) on fibrinolytic activity and inflammatory response in an enhanced recovery after surgery (ERAS) program and to assess the efficacy and safety of Ⅳ-TXA.Methods One hundred and forty-one severe knee osteoarthritis patients following primary unilateral TKA from January 2016 to April 2017 were investigated retrospectively.The patients were divided into following three groups based on the dosage of Ⅳ-TXA after surgery:40 patients received ≤3 g Ⅳ-TXA after surgery (group T1),50 patients received 4 g (group T2) and the other 51 patients received ≥5 g Ⅳ-TXA (group T3).The total blood loss (TBL),hidden blood loss (HBL),transfusion rate,maximum hemoglobin (Hb) drop,the incidence of intramuscular venous thrombosis,deep vein thrombosis (DVT) and pulmonary embolism (PE),fibrinolysis parameters [fibrin(-ogeu) degradation products (FDP),D-dimer],and inflammation markers [C-reactive protein (CRP),interleukin-6 (IL-6)] during perioperative period were evaluated.In addition,correlation analyses between the dosage of Ⅳ-TXA and fibrinolysis parameters and inflammation markers were conducted.Results The mean TBL,HBL and maximum Hb drop in group T2 (537.16±270.43 ml,431.36±271.99 ml,19.68± 10.68 g/L) and T3 (541.31±290.00 ml,439.94±290.71 ml,20.24±8.48 g/L) were lower than those in group T1 (748.22±394.34 ml,P=0.012,0.013;636.47±388.14 ml,P=0.011,0.015;25.88± 11.77 g/L,P=0.005,0.010,respectively).No patient needed transfusion in all groups.There was no statistically difference in the incidence of intramuscular venous thrombosis of lower limbs among three groups (P> 0.05).No episode of DVT or PE occurred in any group in two weeks after surgery.There were negative correlation between the dosage of Ⅳ-TXA and FDP at postoperative day 1 and day 3 (r=-0.191,P=0.025;r=-0.291,P=0.001) and D-dimer on postoperative day 3 (r=-0.176,P=0.048).Moreover,the CRP (r=-0.184,P=0.036) and IL-6 (r=-0.269,P=0.002) level in serum on postoperatire day 1 also showed a negative relationship with the dosage of Ⅳ-TXA after surgery.Conclusion The multiple Ⅳ-TXA (≥4 g) after surgery can further reduce the TBL,HBL and maximum Hb drop following primary TKA in ERAS program without increasing the risk of thrombotic events.Most importantly,the effect of anti-fibrinolysis will be enhanced and may have an anti-inflammatory effect with the dosage of Ⅳ-TXA increased.
9.The efficacy and safety of postoperative retransfusion drain following total hip arthroplasty: a Meta-analysis.
Jinwei XIE ; Bin XU ; Pengde KANG ; Zongke ZHOU ; Bin SHEN ; Jing YANG ; Fuxing PEI
Chinese Journal of Surgery 2016;54(2):108-113
OBJECTIVETo investigate the effectiveness and safety of post-operative retransfusion drain(PRD) after total hip arthroplasty.
METHODSA systematic literature review based on PubMed, EMBase, the Cochrane Library, China Biology Medicine disc, CNKI, VIP and WanFang Database in any language regarding PRD following total hip arthroplasty was performed.The data was evaluated using modified Jadad score and then analyzed using RevMan 5.2.
RESULTSNine randomized controlled trials totaling 1 824 patients, 913 patients in PRD group and 911 in control group, were eligible for data extraction and Meta-analysis.The results indicated that the use of PRD could reduce the requirement of allogeneic blood transfusion when compared with ordinary vacuum drainage (RR=0.61, 95% CI= 0.47-0.79), but the benefit was not found when compared with no drainage group(RR=1.07, 95% CI=0.67-1.71). And the postoperative hemoglobin level was higher in PRD group(MD=0.14, 95% CI=0.01-0.27, P=0.04). No significant difference was identified regarding transfusion index, length of hospital stay, the incidence of febrile reaction and wound-related complications.
CONCLUSIONSPRD in reducing requirement of blood transfusion following THA is effective and safe when compared with ordinary vacuum drainage, but the benefit is not found when compared with no drainage.And more robust evidence is needed to confirm this result.
Arthroplasty, Replacement, Hip ; Blood Transfusion ; China ; Drainage ; methods ; Humans ; Length of Stay ; Postoperative Period ; Randomized Controlled Trials as Topic
10.TWS119 upregulates CCR5 expression of γδT cells by inhibiting STAT3 phos-phorylation
Jing XU ; Leiqing SUN ; Yongqiang CHEN ; Lu ZHENG ; Xiaoting LV ; Fuxing CHEN ; Junquan LIU ; Zhonghai ZHOU
Chinese Journal of Immunology 2016;32(6):825-827,837
Objective:To investigate the mechanisms of TWS119 induced CCR5 expression in hunman γδT cells. Methods:After treatment with various concentrations of TWS119 for 48h, the expression of CCR5 in γδT cells were detected by flow cytometry. The p-STAT3 and GAPDH expression were examined by Western blot analysis. Results: TWS119 could upregulate the expression of CCR5 in dose dependent manner. Western blot analysis revealed that TWS119 inhibit phosphorylation of STAT3,but had no significant impact on GAPDH. In addition, pretreatment of γδT cells with 0. 5 μmol/L STAT3 specific phosphorylation inhibitor Stattic could upregulate the expression of CCR5 and enhance the TWS119 induced CCR5 expression. Conclusion: TWS119 could upregulate CCR5 expression of γδT cells by inhibiting STAT3 phosphorylation in vitro.

Result Analysis
Print
Save
E-mail