Objective To observe the effect of magnesium isoglycyrrhizinate on the expressions of UGT1A, MRP2 protein and mRNA of L-02 cells damaged by triptolide, and to investigate hepatoprotective mechanism of magnesium isoglycyrrhizinate in terms of drug metabolism. Methods L-02 cells were divided into 4 groups:normal group, triptolide group, magnesium isoglycyrrhizinate group and rifampicin group. Magnesium isoglycyrrhizinate group and rifampicin group were pretreated by magnesium isoglycyrrhizinate and rifampicin for 24 h and the remaining two groups added medium. Triptolide were added for 18 h except normal group. Cell survival rate was tested by MTT. The expression levels of UGT1A, MRP2 protein and mRNA were detected by Western blot assay and RT-PCR. Results Compared with triptolide group, cell survival rate was significantly higher in magnesium isoglycyrrhizinate group (P<0.05). Meanwhile, the expression levels of UGT1A, MRP2 protein and mRNA were significantly lower in triptolide group compared with those of control group (P<0.05). The expression levels of UGT1A, MRP2 protein and mRNA were significantly up-regulated in magnesium isoglycyrrhizinate pretreatment group than those of triptolide group (P<0.05). The UGT1A protein and mRNA expressions were significantly decreased in rifampicin pretreatment group than those of magnesium isoglycyrrhizinate group ( P<0.05), but there were no significant differences in MRP2 protein and mRNA expressions between the two groups. Conclusion Magnesium isoglycyrrhizinate shows protective effects on triptolide induced L-02 cell injury, which may be involved with the activation of UGT1A and MRP2.

Objective To observe the sleeping time and analyze its impact on the psychological state and quality of life of patients with type 2 diabetes .Methods Convenience sampling was used to recruit 365 patients with type 2 diabetes in Jiangsu Province Hospital on Integration of Chinese and Western Medicine Affiliated to Nanjing University of Chinese Medicine between April 2014 and April 2015 .All the patients were divided into two groups according to the six-hour cut-off point of sleeping time .They were investigated by means of World Health Organization Quality of Life-BREF ( WHOQOL-BREF), Diabetes Distress Scale ( DDS) and General Self-Efficacy Scale ( GSES).Data were analyzed by t-test, chi-square test, Mann-Whitney U test and multivariate regression analysis .Results The average sleeping time was 7.03 h, and the sleeping time of 109 patients was less than 6 h.Patients with less sleeping time had lower quality of life and self-efficacy scores as well as higher diabetes distress scores than those with sleeping time more than 6 h.The differences were statistically significant (P<0.05).Regression analysis showed that sleeping time was the factor of quality of life (β=0.117, P=0.047), self-efficacy (β=0.136, P=0.024) and diabetes distress(β=-0.118, P=0.046).Conclusion Sleeping time affects the psychological state and quality of life of patients with type 2 diabetes.The medical should pay more attention to the quality of sleep of such patients .

OBJECTIVE: In order to investigate the interaction between the cytokines and keratinocyte and determine the role of cytokines in hyperproliferative of chronic otitis media with cholesteatoma, we observe the expression of matrix metalloproteinase 9 (MMP9), vascular endothelial growth factor (VEGF), keratinocyte growth factor (KGF) and its receptor (KGFR) in middle ear cholesteatoma. METHOD: We examined the expression of MMP9, VEGF, KGF, KGFR and Ki-67 by immunohistochemistry in 50 specimens from chronic otitis media with cholesteatoma and 15 specimens from the normal skin of external auditory meatus. Ki-67 as an evaluation of cholesteatoma proliferation markers were used to detect the keratinocyte proliferative activity. RESULT: (1) The expression of VEGF and MMP9 in cholesteatoma specimens was higher than normal skin, and the difference was statistically significant (t = 4.914, P < 0.01; t = 3.284, P < 0.01). (2) The expression of KGF and KGFR in middle ear tissues was higher than normal skin, and the difference was statistically significant (t = 4.814, P < 0.01; t = 3.104, P < 0.01); The expression of KGF and KGFR increased, and the expression of Ki-67 also correspondly increased in the cholesteatoma. (3) In the tissue MMP9 and VEGF were positive. Mean optical density increased as well. KGF expression also increased accordingly. CONCLUSION MMP9, VEGF, KGF and KGFR proteins played an important role in hyperproliferation of cholesteatoma tissues. VEGF, MMP9 and KGF had a synergistic effect in hyperproliferation of cholesteatoma tissues.
Cholesteatoma, Middle Ear ; pathology ; Cytokines ; metabolism ; Ear Canal ; metabolism ; Ear, Middle ; metabolism ; Fibroblast Growth Factor 7 ; metabolism ; Humans ; Immunohistochemistry ; Keratinocytes ; cytology ; Ki-67 Antigen ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Otitis Media ; pathology ; Receptor, Fibroblast Growth Factor, Type 2 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

Objective To investigate the effects of tripterygium glycosides combined with methotrexate in treatment of rheumatoid arthritis (RA) curative effect and peripheral blood of patients with IL-6, effects of IL-10 and TNF- alpha. Methods 80 patients with RA from February 2014 to August 2016 in our hospital were retrospectively analyzed. According to the different treatment methods by for the observation group, the control group (40 cases). To compare and analyze the curative effect before and after short-term treatment and treatment of patients in the 2 groups were the main clinical symptoms and signs of change and ESR, RF, CRP, IL-6, TNF- alpha, IL-10 index. Results The total effective rate of observation group (92.50％) was significantly higher than the control group (77.50％), the difference was statistically significant (P<0.05), 2 groups of patients before treatment of main clinical symptoms and signs, there was no significant difference, after treatment, clinical symptoms and signs of 2 groups of patients were compared with those before treatment was statistically significant to alleviate the differences (P<0.05), and the observation group of main clinical symptoms and signs were lower than the control group and the remission (down), the difference was statistically significant (P<0.05).Before ESR, the treatment of 2 patients with RF, CRP, TNF- IL-6, alpha, IL-10 index level, no statistical significance, the 2 groups after treatment in patients with ESR, RF, CRP, IL-6, TNF- alpha, IL-10 index level than before treatment with significant difference (P<0.05), and the observation group ESR, RF, CRP, TNF- alpha, IL-6 average index of water compared with the control group decreased significantly, IL-10 index increased significantly than the control group. The difference was statistically significant (P<0.05). Conclusion Tripterygium glycosides combined with methotrexate in treatment of rheumatoid arthritis And it can effectively inhibit the expression of peripheral serum L-6 and TNF- alpha, promote the expression of IL-10 and alleviate the inflammatory reaction

AIM:Atherosclerotic calcification is highly linked with plaque instability and cardiovascular events .Adenosine monophosphate-activated protein kinase ( AMPK) has been involved in the pathogenesis of various cardiovascular disease .The contributions of AMPKαsubunits to the development of atherosclerotic calcification in vivo remained unknown .We hypothesized that AMPKαsubunits may play a role in the development of atherosclerotic calcification .METHODS: Atherosclerotic calcification was generated by 24-week fed of western diet in ApoE-/-background mice .Calcification was evaluated in aortic roots and innominate arteries of ApoE-/-mice or in mice with dual deficiencies of ApoE and AMPKαsubunits globally ( AMPKα1 and AMPKα2 ) , or vascular smooth muscle cell ( VSMC)-specific or macrophage-specific knockout of AMPKα1 with atherosclerotic calcification pone diet . The mechanism of AMPKα1 in regulating Runx2 was further explored in human aortic VSMC .RESULTS: Ablation of AMPKα1 but not AMPKα2 in ApoE-/-background promoted atherosclerotic calcification with increased Runt -related transcription factor ( Runx2 ) expression in VSMC compared with ApoE-/-mice.Conversely, chronic administration of metformin, which activated AMPK, markedly reduced ath-erosclerotic calcification and Runx2 expression in ApoE-/-mice but had less effects in ApoE-/-/AMPKα1 -/-mice.Furthermore, VSMC-but not macrophage-specific deficiency of AMPKα1 in ApoE-/-background promoted atherosclerotic calcification in vivo com-pared with the controls .AMPKα1 silencing in human aortic VSMC prevented Runx 2 from proteasome degradation to trigger osteoblastic differentiation of VSMC .Conversely , activation of AMPK led to Runx 2 instability by inducing its small ubiquitin-like modifier modifi-cation (SUMOylation).Protein inhibitor of activated STAT-1 (PIAS1), the SUMO E3-ligase of Runx2, was directly phosphorylated by
AMPKα1 at serine 510, to enhance its SUMO E3-ligase activity.Ablation of PIAS1 serine 510 phosphorylation inhibited metformin-in-duced Runx2 SUMOylation, and subsequently prevented the effect of metformin on reducing oxLDL-triggered Runx2 expression in hu-man aortic VSMC.CONCLUSION:Deficiency of AMPKα1 in VSMC increases Runx2 expression and promotes atherosclerotic calcifi-cation in vivo.AMPKα1 phosphorylates PIAS1 to enhance Runx2 SUMOyalation and subsequent degradation .

ObjectiveTo investigate the influence of cobalt chloride ( CoCl2 )-mimetic hypoxia on theproliferation,apoptosis and migration of human pancreatic cancer cell fine PANC1.MethodsPANC1 cells were treated with 0(control),100,200,400,800 μmol/L CoCl2 respectively for 24 h.Real-time RT-PCR and Western blot were used to determine hypoxia induced factor ( HIF)-1o mRNA and protein expression respectively,and cell counting kit-8(CCK-8) assays,flow cytometry and cell scratch test were used to examine the proliferation,apoptosis and migration of PANC1 cells,respectively.ResultsIn the control group and 100,200,400 and 800 μmol/L CoCl-2 groups,the expressions of HIF-1t mRNA were 1,1.08 ±0.12,1.12 ± 0.09,1.04±0.11,0.66 ±0.07,and the expressions of VEGF mRNA were 1,2.69±0.35,4.81 ±0.54,2.19 ± 0.21,0.79 ± 0.08,while the expressions of HIF-1 α protein were 0.23 ± 0.03,0.36 ± 0.04,1.15 ± 0.11,1.08 ± 0.09,0.44 ± 0.04; and the expressions of VEGF protein were 0.14 ± 0.02,0.12 ± 0.01,0.95 ±0.09,0.87 ±0.09,0.55 ±0.06; and cell viability rates were 100％,(98.43 ±2.88)％,(76.15 ± 0.70)％,(53.87 ±0.77)％,(35.23 ±0.67)％ ; while cell apoptotic rates were (5.2 ±1.12)％,(5.74 ± 1.07)％,(6.82 ± 1.85)％,(12.09 ±3.53)％,(31.88 ±6.95)％ ; the cell migration distance of PANC1 cells were (43.24 ±3.67)％,(59.46 ±5.39)％,(80.56 ±8.05)％,(63.89 ±5.96)％,(9.09 ± 1.59 ) ％.Compared with those of control group,the expressions of VEGF mRNA,VEGF and HIF-1 α protein,cell migration distance showed a two-way variation ( ascending first and descending later) (P ＜0.05 ),and the expression of HIF-1α mRNA and cell proliferation rate was decreased in a dose-dependent manner,while the cell apoptosis was increased in a dose-dependent manner.Conclusions CoCl2 significantly inhibits the proliferation and promotes apoptosis of PANC1 cells at certain level.CoCl2 has a two-way effect on the migration of PANC1 cells,and it may be related to the direct injury of high concentration of CoCl2 on cells.

OBJECTIVE:To investigate the effects of Glycyrrhiza uralensis extract (GE) on the expression of uridine diphosphate glucuronyltransferase 1A (UGT1A) and multidrug resistance associated protein 2 (MRP2) in human liver L-02 cells damaged by triptolide (TP),and to study attenuated mechanism of G.uralensison for TP.METHODS:The survival rates of L-02 cells were determined by MTT assay after cultured with 0 (blank control),40,80,160 nmol/L TP for 12,18,24 h.L-02 cells were divided into blank control group (blank culture medium),model control group (80 nmol/L TP) and GE pretreatment group (adding 80 nmol/ L TP after pretreated with 30,60,90 mg/L GE for 24 h);after cultured for 18 h,survival rates of L-02 cells were determined by MTT assay.Rifampin (RIF) group (positive control,adding 80 nmoi/L TP after pretreated with 10 μmol/L RIF for 24 h) was added on the basis of the above grouping (GE concentration of 60 mg/L in GE pretreatment group).After cultured for 24 h,the protein expressions of UGT1A and MRP2 were detected.RESULTS:The inhibition effect of TP on cell proliferation was positively correlated with the concentration and the time.Compared with blank control group,cell survival rate of model control group was decreased significantly (P＜0.05),and the protein expression of MRP2 was decreased significantly (P＜0.01).Compared with model control group,cell survival rates of 30,60,90 mg/L GE pretreatment groups were all increased significantly (P＜0.01).The protein expressions of UGT1A and MRP2 were increased significantly in 60 mg/L GE pretreatment group (P＜0.01).CONCLUSIONS:GE pretreatment can relieve TP-induced human liver L-02 cell damage,and its attenuated mechanism may be associated with the increase the expression of UGT1A and MRP2.

Objective:To investigate the diagnostic value of CT-guided puncture biopsy combined with serum gamma-glutamyltransferase (GGT) and abnormal prothrombin (PIVKA-Ⅱ) in serum alpha-fetoprotein(AFP) negative primary liver cancer (PHC).Methods:Eighty patients with AFP negative PHC treatment in Fuyang Women and Children′s Hospital from January 2018 to March 2021 were selected as AFP negative PHC group, and another 85 patients diagnosed with benign liver tumor during the same period were selected as the control group retrospectively. The patients of the two groups underwent CT-guided biopsy and the levels of GGT and PIVKA-Ⅱ were detected. The single diagnostic value and combined diagnostic value of AFP negative PHC were analyzed by receiver operating characteristic (ROC) curve.Results:Seventy-five of the 80 patients in the AFP negative PHC group were confirmed as liver malignant lesions by biopsy, with a coincidence of 93.75%, and 84 of the 85 patients in the control group were confirmed as liver benign lesions by biopsy, with a coincidence of 98.82%. The levels of AFP, GGT and PIVKA-Ⅱ in AFP negative PHC group were significantly higher than those in the control group: (175.67 ± 39.58) μg/L vs. (18.74 ± 7.42) μg/L, (1 245.37 ± 255.41) U/L vs. (486.63 ± 89.05) U/L, (385.49 ± 30.27) AU/L vs. (25.84 ± 7.66) AU/L, there were statistical differences ( P<0.05). Spearman correlation analysis showed that serum AFP was positively correlated with GGT and PIVKA-Ⅱ ( r = 0.858 and 0.429, P<0.05). The results of ROC curve showed that the area under curve of CT-guided biopsy combined with GGT and PIVKA-Ⅱ in the diagnosis of AFP negative PHC was 0.877, the sensitivity was 91.19%, the specificity was 87.34%. Conclusions:CT-guided biopsy combined with GGT and PIVKA-Ⅱ detection of AFP negative PHC can effectively improve the diagnostic value.

Objective:To explore the diagnostic value of contrast-enhanced mammography (CEM) and MRI in differentiating benign and malignant breast lesions based on the 2013 breast imaging reporting and data system (BI-RADS) lexicon and the supplement on CEM.Methods:The clinical and imaging data of 83 patients with breast lesions from March 2019 to April 2022 in the Third Affiliated Hospital of Soochow University were retrospectively analyzed. Totally 100 breast lesions from 83 female patients aged 28 to 78 (49±14) years, were divided into benign lesions (50 lesions) and malignant lesions (50 lesions) according to the pathological results. The t-test, χ 2 test and Fisher′s exact test were used to compare the differences of clinical and imaging features between benign and malignant lesions, and these imaging features which had statistical differences were established CEM and MRI models by multivariate logistic regression analysis respectively. The receiver operating characteristic curves and the area under the curve (AUC) were used to assess the diagnostic efficacy of two models in differentiating benign and malignant breast lesions. Using the DeLong test compared the AUC. Results:Multivariate logistic regression analysis showed that associated features (OR=9.075,95%CI 1.430-57.570, P=0.019), lesion conspicuity (OR=6.180,95%CI 2.608-14.646, P<0.001), mass margin (OR=2.193,95%CI 1.405-3.422, P=0.001) and calcification distribution (OR=2.147,95%CI 1.157-3.986, P=0.015) were independent predictors of differentiating benign and malignant breast lesions in CEM, and then the predictive model of CEM was constructed. Time-signal intensity curve (OR=9.230, 95%CI 3.178-26.805, P<0.001), associated features (OR=5.289,95%CI 1.343-20.831, P=0.017) and mass margin (OR=2.192,95%CI 1.336-3.597, P=0.002) were independent predictors of differentiating benign and malignant breast lesions in MRI, and the predictive model of MRI was constructed. The AUC of CEM and MRI models for differentiating benign and malignant breast lesions were 0.947 and 0.930 respectively, and two models were no significant difference ( Z=0.68, P=0.494). Conclusion:The diagnostic efficacy of CEM and MRI in differentiating benign and malignant breast lesions is comparable based on the 2013 BI-RADS lexicon and the supplement on CEM.

Objective To summarize the clinical characteristics of Beh?et's disease (BD) in children with gastrointestinal involvement. Methods We retrospectively analyze the children BD with gastrointestinal involvement who were diagnosed in our hospital in recent 10 years. Results Twenty-two children were identified. The average age of onset was(6.1±4.0) years. The time from disease onset to clinical diagnosis was (1.2±2.1) years on average. Fifteen children had abdominal pain, diarrhea and hematochezia. Seven cases had positive endoscopic findings without any gastrointestinal symptoms. Twenty cases received corticosteroids therapy, 13 cases of them were treated with Cyclophosphamide/Methotrexate (CTX/MTX), 3 refractory cases were treated with biologics. Patients were followed up for (28±32) month on average. Eight patients' condition was stable, 7 patients were refractory, 3 patients died, 4 patients were lost to follow-up. At the same term, 5 patients without gastrointestinal involvement who received corticosteroids and CTX/MTX therapy were stable. Conclusion It is difficult to diagnose children BD at early stage. Gastrointestinal involvement may not be found, while the gastrointestinal endoscopy is of great importance in the diagnosis of the disease. Gluco-corticoid combined with immunosuppressive agents are effective. As to refractory patients, biological agent might be used although the recurrence is common. Compared with BD without gastrointestinal involvement, children BD with gastrointestinal involvement have serious condition and poor prognosis.