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ObjectiveTo investigate the role and mechanism of hyodeoxycholic acid （HDCA） in the progression of metabolic associated fatty liver disease （MAFLD）， and to provide a new theoretical basis for further clarifying the pathogenesis of MAFLD. MethodsL02 hepatocytes were used as experimental cells， and palmitic acid was used to induce steatosis in L02 cells. The farnesoid X receptor （FXR） siRNA interference chain technique was used to construct a hepatocyte cell line with low FXR expression. CCK8 assay was used to observe the effect of HDCA on L02 steatosis hepatocytes at different concentrations （0， 100， 200， 300， and 400 μmol/L） and time points （12， 24， 36， and 48 hours）. The method of qRT-PCR was used to measure the mRNA expression levels of FXR， proliferating cell nuclear antigen （PCNA）， Cyclin D1， phosphatidylinositol 3-kinase （PI3K）， and protein kinase-B （AKT）， and Western blot was used to measure the protein expression levels of FXR， Cyclin D1， PCNA， PI3K， phosphorylated PI3K （p-PI3K）， AKT， and phosphorylated （p-AKT）. A one-way analysis of variance was used for comparison of normally distributed continuous data with homogeneity of variance between multiple groups， and the Tukey HSD test was used for further comparison between two groups； the Welch analysis of variance was used for comparison of normally distributed continuous data with heterogeneity of variance between multiple groups， and the Games-Howell test was used for further comparison between two groups. The independent-samples t test was used for comparison between two groups. ResultsCCK8 assay showed a significant reduction in the viability of L02 cells and steatosis hepatocytes treated by 300 μmol/L HDCA （P<0.05）， and qRT-PCR showed a significant increase in the mRNA expression level of FXR and significant reductions in the mRNA expression levels of PCNA， Cyclin D1， PI3K， and AKT （all P<0.05）. Western blot showed a significant increase in the protein expression level of FRX （P<0.05）， and after interference of FXR expression in L02 cells， there were significant increases in the protein expression levels of PCNA， PI3K， p-PI3K， AKT， and p-AKT （all P<0.05）. ConclusionHDCA inhibits the PI3K/AKT signaling pathway by upregulating FXR expression， thereby inducing a reduction in the viability of steatosis hepatocytes.
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Objective:To summarize the clinical characteristics of neonatal cerebral infarction and its risk factors, so as to provide a reference for clinical diagnosis and early prevention of the disease.Methods:This study retrospectively analyzed the demographic data, clinical manifestations and brain imaging features of neonates with cerebral infarction ( n=45) admitted to the Department of Neonatal Critical Care Medicine of the Affiliated Children's Hospital of Xi'an Jiaotong University from June 2012 to July 2020. Ninety newborns without cerebrovascular disease matched for date of birth and gestational age were selected as the control. Two independent sample t-test, rank-sum test, Chi-square or corrected Chi-square test were used for univariate analysis and binary logistic regression were applied for analyzing the risk factors for neonatal cerebral infarction. Results:A total of 45 infants with clinically diagnosed neonatal cerebral infarction were enrolled, including eight small for gestational age and three macrosomia infants. The median age at disease onset was 1 d (1-2 d). There were 71% (32/45) presenting with convulsions as the first symptom, 4% (2/45) admitted with apnea and respiratory distress as the chief complaints, respectively,11% (5/45) having poor response and 9% (4/45) showing no obvious clinical manifestations. Cranial MRI and magnetic resonance angiography identified left hemisphere lesion in 25 cases (56%), right hemisphere lesion in 16 (36%) and both in four (9%). Thalamus and basal ganglia were involved in 11 cases. The lesions were supplied by middle cerebral artery [38% (17/45)], anterior cerebral artery ( n=1), posterior cerebral artery ( n=4), anterior and middle cerebral arteries ( n=4), middle and posterior cerebral arteries ( n=16), or anterior, middle and posterior cerebral arteries ( n=3). Univariate analysis showed that the proportions of small for gestational age [18% (8/45) vs 6% (5/90), χ 2=5.15], cesarean section after failure of trial of labor [18% (8/45) vs 1% (1/90), χ 2=10.85], meconium stained amniotic fluid [33% (15/45) vs 9% (8/90), χ 2=12.68], fetal distress [20% (9/45) vs 3% (3/90), χ 2=8.34] and neonatal asphyxia [16% (7/45) vs 2% (2/90), χ 2=6.56] were all higher in the infarcted infants than those in the control (all P<0.05). Binary logistic regression analysis revealed that small for gestational age ( OR=3.981, 95% CI: 1.075-14.742, P=0.039), cesarean section after failure of trial of labor ( OR=17.959, 95% CI: 2.032-158.698, P=0.009) and fetal distress ( OR=5.756, 95% CI: 1.129-29.331, P=0.035) were independent risk factors for neonatal cerebral infarction. Conclusions:Most neonates with cerebral infarction would have convulsions initially, while some are asymptomatic. Middle cerebral arteries are often involved in the lesion. The risk of this disease may be increased in small for gestational age infants, cesarean section after failure of trial of labor and fetal distressed cases.
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Objective:A clinical prediction model was constructed based on the related factors affecting neonatal early-onset sepsis (EOS).Methods:A retrospective study was conducted. The patients with EOS amditted to the neonatal intensive care unit of Children′s Hospital Affiliated to Xi′an Jiaotong University from April 2015 to April 2020 were enrolled. The demographic data and the clinical indicators within 8 hours after admission were collected. The death 7 days after admission was taken as the end event. The differences of various indexes between the survival group and the death group were compared. After univariate analysis of the indexes that may have an impact on the prognosis, binary logistic regression analysis was performed; The predictive model was established for the factors that may affect the prognosis; the predictive value of the relevant models was analyzed by recevier operating characteristic (ROC) curve, and the model was verified by independent clinical medical records.Results:A total of 139 children were enrolled, and 41 died within 7 days, with a fatality rate of 29.50%. Compared with the survival group, the dead group had higher white blood cells (WBC), serum procalcitonin (PCT), lactic acid (Lac), creatinine (Scr), D-dimer and Paediatric Risk of Mortality (PRISM) score {WBC(×10 9/L): 24.15[4.36, 29.36] vs 21.21[19.14, 28.36], PCT: (67.32±40.36)ng/L vs (37.76±25.11)ng/L, Lac: (8.69±6.17)mmol/L vs (2.34±1.11)mmol/L, Scr: (239.99±68.46)μmol/L vs (65.31±34.34)μmol/L, D-dimer(mg/L): 5.21[2.06, 21.49] vs 0.34[0.26, 0.45], PRISM Ⅲ: (19.52±6.25)s vs (10.63±2.05)s, all P<0.05}, and lower fibrinogen (Fib), platelet count (PLT) and hemoglobin concentration (Hb) [Fib: (1.48±1.19)g/L vs (2.44±0.83)g/L, PLT: (154±58)×10 9/L vs (189±29)×10 9/L, Hb: (169±49)g/L vs (182±52)g/L, all P<0.05]. The incidence of placental/umbilical cord lesions, amniotic fluid pollution, asphyxia, premature delivery, premature rupture of membranes, positive etiology and maternal infection in the death group were higher than those in the survival group, while the gestational age and weight were lower than those in the survival group (all P<0.05); Binary logistic regression analysis showed that Lac, PCT and premature rupture of membranes were independent risk factors for the prognosis of EOS [odds ratio ( OR) and 95% confidence interval (95% CI): Lac was 1.23(1.00-2.05), PCT was 1.05(1.03-1.85), premature rupture of membranes was 2.59(1.89-3.32), all P<0.05]; ROC curve analysis showed that the area under the curve (AUC) of the prediction model was 0.967; the predicted sensitivity was 88.70%; and the specificity was 78.20% respectively. Conclusions:PCT, Lac and premature rupture of membranes are independent risk factors affecting the prognosis of EOS. The clinical prognosis prediction model constructed by combining PLT, gestational age and weight has good prediction efficiency.
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Objective:To explore the effectiveness and safety of plasma exchange in the treatment of neonatal extremely severe hyperbilirubinemia.Methods:A retrospective analysis was performed on the data of 18 cases, who were all newborns with extremely severe hyperbilirubinemia and treated with plasma exchange in the NICU at Xi ′an Children′s Hospital from April 2019 to December 2019.The changes of serum total bilirubin, indirect bilirubin, albumin, white blood cells, red blood cells, platelets, hematocrit, hemoglobin, serum sodium, serum potassium, serum calcium, blood glucose, blood coagulation and mean arterial pressure were compared before and after plasma exchange.Results:A total of 18 eligible children were included.The peak value of total bilirubin was (571.2±113.3) μmol/L before treatment, and the value after treatment was (235.8±66.7) μmol/L, whose difference was statistically significant ( P<0.05). The exchange rate of bilirubin was (58.5±8.4)%.There were no statistically significant differences in the changes of white blood cells, platelets, hemoglobin, hematocrit, serum sodium, serum potassium, serum chloride, serum calcium, serum glucose, and albumin before and after plasma exchange (all P>0.05). There were no allergic reactions, hypotension, plasma separator or pipeline coagulation and other adverse reactions during plasma exchange. Conclusion:Plasma exchange therapy can remove serum bilirubin quickly, effectively and safely, and may be a new treatment for neonatal hyperbilirubinemia.
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Objective:To explore the expression and clinical significance of IL-1β and IL-1β receptor antagonist(IL-1ra)in persistent pulmonary hypertension of the newborn(PPHN)secondary to sepsis.Methods:The newborns with sepsis were enrolled in the Department of Neonatal Intensive Care Unit(NICU)of Xi′an Children′s Hospital from January 2018 to November 2020.The newborns with sepsis were divided into two groups: the newborns without PPHN( n=108)were the control group and the newborns with PPHN( n=44)were the experimental group.Clinical data, laboratory examination and bedside echocardiography of all the newborns were collected to analyze the differences between the two groups.The expression levels of IL-1β and IL-1ra in neonatal plasma of the two groups were detected by enzym-linked immunosorbination(ELISA), and their roles in neonatal sepsis with PPHN were further analyzed.The risk factors of neonatal sepsis with PPHN were analyzed by multivariate Logistic regression, and the early prediction value of the risk factors for neonatal sepsis with PPHN were evaluated by the receiver operating characteristic(ROC)curve. Results:There were no significant differences in gestational age[(39.11±0.55)w vs(38.85±0.72)w], birth weight[(3.30±0.49)kg vs(3.24±0.55)kg]and proportions of males[60(55.6%)vs 30(68.2%)]between the two groups( P>0.05). The right ventricular diameter[(9.57±0.35)mm], pulmonary artery pressure[(51.36±5.91)mmHg]and the level of N-terminal brain natriuretic peptide(NT-proBNP)[(25436.83±12343.18)ng/L)]significantly increased in the experimental group than those in the control group[(8.77±0.41)mm, (31.24±5.11)mmHg, (11267.09±4405.48)ng/L, respectively, P<0.05]. Before treatment, the expression levels of plasma IL-1β[(31.24±5.25)ng/L]and IL-1ra[(41.94±10.13)ng/L]in the experimental group were significantly higher than those in the control group[(18.27±4.47)ng/L, (21.47±8.76)ng/L, respectively, P<0.05]. The expression levels of plasma IL-1β[(10.46±3.17)ng/L]and IL-1ra[(10.58±2.94)ng/L]in the experimental group after treatment were significantly lower than those before treatment[(31.24±5.25)ng/L, (41.94±10.13)ng/L , respectively, P<0.05]. Multivariate Logistic regression analysis showed that IL-1β and NT-proBNP were the independent risk factors for neonatal sepsis with PPHN( P<0.05). ROC curve analysis showed that IL-1β and NT-proBNP had the good predictive value for the occurrence of neonatal sepsis with PPHN( P<0.05). IL-1β combined with NT-proBNP has the better predictive value for neonatal sepsis with PPHN. Conclusion:IL-1β combined with NT-proBNP have the high predictive value for PPHN of the newborns secondary to sepsis.
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ObjectiveTo obtain HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect by transfecting HSC-T6 cells with CRISPR/Cas9 lentiviral vector, and to provide a good method for further functional research and new strategies for the clinical treatment of liver fibrosis. MethodsThe COX-2 gene-specific sgRNAs (COX-2-sgRNA-1, COX-2-sgRNA-2, COX-2-sgRNA-3) were designed, synthesized, and connected to the GV371 vector, and the recombinant plasmid and the packaging plasmid were transfected into 293T cells to form lentivirus particles; the fluorescence method was used to measure virus titer. The most appropriate amount of the virus was calculated based on MOI. Lenti-Cas9-puro was transfected into HSC-T6 cells, and HSC-T6-Cas9 cells were screened out by puromycin; Lenti-COX-2-sgRNA-EGFP was transfected into HSC-T6-Cas9 cells to obtain HSC-T6-COX-2-/- cells. Cruiser enzyme digestion and Western blot were used to verify gene knockout at the gene and protein levels. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsSequencing verified that the COX-2-sgRNA expression vector was constructed successfully. Recombinant expression plasmids and packaging plasmids were transfected into 293T cells to form lentivirus particles, and the fluorescence method showed a virus titer of ＞1×108. HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect were successfully constructed. The HSC-T6-Cas9 group had significantly higher relative mRNA expression of LV-Cas9-Puro than the CON group (541.93±105.76 vs 1.00±0.02, t=12.995, P＜0.01). Cruiser enzyme digestion and Western blot showed that the CRISPR/Cas9 lentivirus expression vectors played a role in the target, among which COX-2-sgRNA-2 knockout had the most significant effect, and this group had a significant reduction in the protein expression level of COX-2 compared with the CON group and the NC group (both P＜0.05), suggesting that COX-2-sgRNA was active. ConclusionA CRISPR/Cas9 lentivirus vector is successfully constructed for COX-2 target gene, and HSC-T6-COX-2-/- cells with stable COX-2 gene knockout are obtained.
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Objective:To investigate the clinical characteristics of neonatal congenital tongue base cyst.Methods:This retrospective study involved 35 neonates with congenital tongue base cyst diagnosed in the neonatal intensive care unit (NICU) of Xi'an Children's Hospital from June 2013 to December 2019. General information, clinical manifestations, supplementary results, treatment and prognosis of these babies were described.Results:(1) The median age at the onset of the disease was 12.5 (0~28) d and the median age at admission was 15 (0~28) d for these babies. The main clinical manifestations were laryngeal stridor (28/35, 80.0%), inspiratory dyspnea and crying, especially when feeding (26/35, 74.3%) and choking and spitting with feeding (23/35, 65.7%). (2) Among the 35 cases, 15 (42.9%) required emergency endotracheal intubation due to significant dyspnea when were admitted to the NICU and five out of them were considered for having tongue base mass under laryngoscopy, while the other 10 cases underwent bedside electronic laryngoscopy after endotracheal intubation, in which space-occupying lesions were found. Tongue base cyst was considered in seven cases with laryngeal stridor complicated by protracted pneumonia using fiberoptic bronchoscopy. The other 13 cases were examined by electronic laryngoscope and considered as tongue base cyst. Thirty-five cases underwent cervical ultrasound and only five of them were considered as tongue base tumor. Thirty-two cases underwent cervical CT scan and only two of them were normal. Three cases were found to have tongue base cyst by cranial MRI. (3) Thirty-four cases were treated by radiofrequency ablation assisted with self-retaining microlaryngoscope and general anesthesia, while the other one firstly received puncture and drainage under direct laryngoscope due to the difficult intubation because of the huge tongue base cyst and then underwent surgery when stable. Only one case (2.9%) relapsed after surgical treatment during regular follow-up.Conclusions:Neonatal congenital tongue base cyst has an early onset and atypical clinical manifestations. Electronic laryngoscopy/fiberoptic bronchoscopy combined with neck CT or MRI examination should be performed promptly in patients with laryngeal stridor and inspiratory dyspnea to facilitate the accurate diagnosis and timely surgery is required for.
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Objective:To analyze the clinical features of moderate or severe neonatal hemophilia, and improve the understanding of this disease.Methods:Eleven cases of neonates with moderate or severe hemophilia admitted to our NICU from January 2012 to June 2019 were enrolled.The clinical features, laboratory data, treatments and prognosis of these 11 neonates were retrospectively analyzed.Results:All the neonates were male, and were diagnosed hemophilia A. Seven neonates presented with intracranial hemorrhage including one case complicated with cerebral hernia.Only two of these neonates with intracranial hemorrhage had neurological abnormalities.One case presented with right adrenal hematoma, and one case presented with retroperitoneal hematoma.Jaundice was observed in nine cases, and seven cases, jaundice appeared within two days after birth, whose earliest was 12 hours after birth, and the highest total bilirubin was 388 μmol/L.All cases had prolonged activated partial thromboplastin time.All neonates had decreased activity of coagulation factor Ⅷ including eight moderate and three severe neonatal hemophilia A. Four cases had genetic testing.Three cases infused with fresh frozen plasma and cryoprecipitate and the rest treated with coagulation factorⅧ infusion.Ten cases improved after treatment, and one case abandoned treatment.Conclusion:Moderate or severe hemophilia is often complicated with intracranial hemorrhage or abdominal hemorrhage, often accompanied with hyperbilirubinemia.Imaging examination should be performed to exclude occult bleeding, and coagulation function and coagulation factor activity should be detected as soon as possible.Blood coagulation factors are infused according to the different expected factors of different bleeding sites.
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ObjectiveTo investigate the effect and significance of cyclooxygenase-2 (COX-2) inhibitors on the expression of the Acsl gene family in the ileum of rats with nonalcoholic fatty liver disease (NAFLD). MethodsA total of 45 Sprague-Dawley rats were randomly divided into normal control group (15 rats given normal diet), NAFLD model group (15 rats given high-fat diet), and nimesulide group (15 rats given high-fat diet and nimesulide). All rats were sacrificed after 12 weeks of feeding, and then blood samples were collected from the inferior vena cava to measure total cholesterol (TC) and triglyceride (TG). HE staining and oil red O staining were performed for the liver to evaluate the degree of hepatic steatosis in each group, and quantitative real-time PCR was used to measure the mRNA expression of the Acsl family genes in the ileum. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the normal control group, the NAFLD model group had significant increases in serum TC and TG and marked hepatic steatosis (all P＜0.05); compared with the NAFLD model group, the nimesulide group had significant reductions in serum TC and TG and degree of hepatic steatosis (all P＜0.05). Compared with the normal control group, the NAFLD model group had a significant increase in the expression of COX-2 in the ileum (P＜0.05), and compared with the NAFLD model group, the nimesulide group had a significant reduction in the expression of COX-2 in the ileum (P＜005). Compared with the normal control group, the NAFLD model group had significant increases in the mRNA expression of Acsl3 and Acsl5 in the ileum (both P＜0.05), and compared with the NAFLD model group, the nimesulide group had significant reductions in the mRNA expression of Acsl3 and Acsl5 (both P＜0.05). ConclusionThe COX-2 inhibitor nimesulide can regulate the expression of the Acsl gene family in the ileum of rats with NAFLD, suggesting that COX-2 inhibitors may inhibit the progression of NAFLD through the Acsl gene.
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Cell senescence is a basic cell response triggered by various stimuli and is a basic characteristic of living organism. Liver fibrosis refers to the pathological process of diffuse excessive deposition of extracellular matrix in the liver caused by abnormal proliferation of connec-tive tissue due to various pathogenic factors and can progress to liver cirrhosis and liver cancer. Studies have shown that cell senescence is closely associated with the progression of liver fibrosis. This article reviews the regulatory effect of different types of cell senescence on liver fibrosis.
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Objective To evaluate the clinical effect and prognostic factors of patients with liver metastases treated with three dimensional conformal radiation therapy (3-DCRT) with boost radiotherapy.Methods From Jan 2010 to Jan 2016,120 patients of liver metastases treated in the Third Rradiotherapy Department of the Third Affiliated Hospital of Harbin Medical University (all patients were treated for the first time and were not suitable for surgical treatment) were restrospectively analyzed.Results 1,2,3-year survival rates were 74.7％,20.2,13.5％,median survival was 19 months.For univariate analysis,those with less number of extrahepatic organ metastasis,fewer liver metastases,smaller size of liver metastases,higher radiation dose,no chemotherapy before radiation and Karnofsky score ≥80 points were with higher 1-,2-and 3-year survival rate and longer median survival time (P ＜ 0.05).Multivariate analysis showed that the number of involved extrahepatic organs,the maximum diameter of liver metastases,numbers of metastases and radiation dose are independent factors to affect the survival of patients with liver metastases (P ＜ 0.05).Conclusions Patients with liver metastases treated with 3D-CRT and boost radiotherapy have satisfactory local control and 1,2,3-year survival rate as well as lower incidence of toxicity damage to the liver.
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Objective To investigate the safety and effectiveness of a postoperative intravenous injection of Dezocine on the recovery from emergency agitation after Sevoflurane based anesthesia in patients receiving radical resection of rectal carcinoma.Methods A total of 66 patients receiving Sevoflurane Fentanyl anesthesia during radical resection of rectal cancer in our hospital from February 2015 to January 2017 were enrolled.They were randomly divided into a control group (normal saline,n=33) and an observation group (Dezocine,n 33).At 15 minutes before the end of operation,the control group received an intravenously administered normal saline,and the Dezocine group received 0.1 mg/kg dezocine in normal saline.The emergency agitation (EA),incidence of adverse reactions,and hemodynamic indexes were collected in both groups.Results The mean values of arterial blood pressure (MAP) and heart rate (HR) were significantly lower in the observation group than in control group at all four observation time points (tMAP =3.228,3.603,5.431,5.568;tHR =3.447,3.739,4.425,4.476;all P ＜0.05).The incidence of EA was significantly lower in the observation group (6.1％) than in the control group (24.2％) (x2=4.242,P=0.039).The stay time in post anesthesia care unit (PACU) was significantly shorter in the observation group [(54 ± 11) min] than in the control group [(72± 12) min] (t =9.317,P =0.000).In addition,the observation group had lower agitation scores and lower Ramsay sedation scores compared with the control group at all time points during extubation (tagitation =2.862,3.348,3.411,3.159;tRamsay =3.508,3.617,3.207,2.931;all P＜0.05).Conclusions The use of Dezocine during anesthesia recovery period in patients undergoing radical resection of rectal cancer has an analgesic effect.h can effectively reduce the incidence of EA and maintain hemodynamic stability.
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Objective:To establish a method for simultaneously determining 3 components ( hesperidin, magnolol and honokiol) in Xiaoji Huachong powder by UPLC. Methods: The experiments were performed on a Thermo Accucore C18 column ( 150 mm × 4. 6 mm, 2. 6 μm). The mobile phase was methanol-0. 5% glacial acetic acid in water with gradient elution, the flow rate was 0. 7 ml ·min-1 , the column temperature was maintained at 30 ℃,the detection wavelength was set at 283 nm and the injection volume was 2μl. Results:The 3 constituents showed a good linear relationship within the range of 4. 707-28. 242 μg·ml-1(r=0. 9994) for hes-peridin, 5. 085-30. 510 μg·ml-1(r=0. 9997) for magnolol and 10. 651-63. 908 μg·ml-1(r=0. 9995) for honokiol. The average recovery was 101. 2%,104. 8% and 99. 4% with the RSDs of 1. 11%,1. 68% and 1. 49% (n=6), respectively. Conclusion: The experiment provides a simple and effective method that can be used to evaluate the quality of Xiaoji Huachong powder.
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Objective To investigate the effects of nonyphenol (NP)exposure during pregnant and lactation period on expression of cytochrome P450 2E1 (CYP2E1) mRNA and protein of hepatic tissues in offspring rats.Methods Pregnant rats were assigned to four groups:the exposure groups received gavage with NP at dose levels of 50 mg · kg-1 · d-1 (low dose group),100 mg · kg-1 · d-1 (medium dose group),200 mg · kg-1 · d-1 (high dose group) and the control group was treated with corn oil alone,NP exposure time was limited from gestational day 6 to postnatal day 21.The newborn rats were sacrificed at 90 days after birth,followed by blood collection and serum separation.Then,serum biochemical indicators of liver function and lipid levels were detected.Glutathione peroxidase (GSH-PX),superoxide dismutase (SOD) activities,malondialdehyde (MDA) level and CYP2E1 mRNA and protein levels were determined in hepatic tissues.Pathologic changes in hepatic tissues were observed with HE staining.Results Compared with the control group,the aspartate aminotransferase (ALT),alanine aminotransferase (AST) levels and AST/ALT ratio of offspring rats in exposure groups were increased in a dose-dependent manner (P＜0.05),as well as serum TG,TC and LDL-C levels were increased (P＜0.05).The liver tissue structure of the control group was normal.The hepatic sinus of the medium dose group was showed mild expansion and inflammatory cellular infiltration.Otherwise,the liver of high dose group had a large amount of lipid droplets.Compared with the control group,SOD and GSH-PX activities were obviously decreased,while MDA level was significantly decreased in each exposure group (P＜0.05).CYP2E1 mRNA and protein expression levels of medium and high dose groups were higher than those in the control group (P＜0.05).Conclusion Exposure to NP during gestation and lactation period can induce lipid metabolism disorders and inflammatory lesions in hepatic tissues of offspring rats,and it maybe associated With up-regulation of CYP2E1 expression.
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Objective To investigate the effects of nonyphenol (NP)exposure during pregnant and lactation period on expression of cytochrome P450 2E1 (CYP2E1) mRNA and protein of hepatic tissues in offspring rats.Methods Pregnant rats were assigned to four groups:the exposure groups received gavage with NP at dose levels of 50 mg · kg-1 · d-1 (low dose group),100 mg · kg-1 · d-1 (medium dose group),200 mg · kg-1 · d-1 (high dose group) and the control group was treated with corn oil alone,NP exposure time was limited from gestational day 6 to postnatal day 21.The newborn rats were sacrificed at 90 days after birth,followed by blood collection and serum separation.Then,serum biochemical indicators of liver function and lipid levels were detected.Glutathione peroxidase (GSH-PX),superoxide dismutase (SOD) activities,malondialdehyde (MDA) level and CYP2E1 mRNA and protein levels were determined in hepatic tissues.Pathologic changes in hepatic tissues were observed with HE staining.Results Compared with the control group,the aspartate aminotransferase (ALT),alanine aminotransferase (AST) levels and AST/ALT ratio of offspring rats in exposure groups were increased in a dose-dependent manner (P＜0.05),as well as serum TG,TC and LDL-C levels were increased (P＜0.05).The liver tissue structure of the control group was normal.The hepatic sinus of the medium dose group was showed mild expansion and inflammatory cellular infiltration.Otherwise,the liver of high dose group had a large amount of lipid droplets.Compared with the control group,SOD and GSH-PX activities were obviously decreased,while MDA level was significantly decreased in each exposure group (P＜0.05).CYP2E1 mRNA and protein expression levels of medium and high dose groups were higher than those in the control group (P＜0.05).Conclusion Exposure to NP during gestation and lactation period can induce lipid metabolism disorders and inflammatory lesions in hepatic tissues of offspring rats,and it maybe associated With up-regulation of CYP2E1 expression.
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Objective To observe the effect of different doses of butorphanol and sufentanil repeatedly epidural injected on the neurobehavior function in bone cancer model rats.Methods A PE-530 catheter was inserted into the epidural space of all male Sprague-Dawley rats(not mated,weighting 150-180 g) at L1-2 level.Three days after operation,64 rats without any motor dysfunction were randomly divided into eight groups (n=8):sham operated group (group C),normal saline with bone cancer pain group (group N),butorphanol groups(group B 1,B2,B3)and sufentanil groups (group S1,S2,S3).Bone cancer pain model was constructed in group N,B and S when rats in group C were sham operated.Rats in group C and N were epidurally injected NS 30μl each,and rats in group B1,B2 and B3 were respectively epidurally injected butorphanol 25,50,100 μg (all diluted to 30 μl with NS),when rats in group S1,S2 and S3 were respectively cpidurally injected sufentanil 1,2,4 μg (all diluted to 30 μl with NS) on time per day for 10-14 days after modeling.The neurobehavior paw withdrawal threshold (MWT) of the left hind claw was recorded to observe the changes in pain behavior.The neurobehavior function of rats were recorded by BBB (BASSO,BEATTIE and BRESNAHAN) score and the inclined plane test.Results Compared with group C((67.65±9.29) g),the MWT of the model groups obviously decreased before the first time of injection (N (15.23± 2.46) g,B 1 (16.14±2.28) g,B2(15.42±3.22) g,B3(14.35±2.32) g,S1 (15.37±2.11)g,S2(15.22±2.93) g,S3(16.25± 2.36) g) (all P＜0.05)).Compared with group N((16.13±2.37) g),the MWT of group B2,B3 and S3 increased obviously after the first time of injection ((35.12±5.16) g,(35.63± 1.53) g and (34.24±5.93) g) (P＜ 0.05).At the first day of injection,there was no significant difference in the BBB scores and the inclined plane test between the model groups (P＞0.05).At 6 h after the forth injection the inclined plane test and the BBB scores of group B3 were obviously decreased compared with group N ((34.72 ± 4.56) ° and (10.64 ± 1.82) points to (43.15±4.67)° and (14.05±1.78) points (P＜0.05)).Conclusion The results provide evidence that repeatedly epidural injection of butorphanol 50 μg or 100 μg or sufentanil 4 μg can reduce the pain of the rats with bone cancer pain.But repeated epidural injection of butorphanol 100 μg can injure the neurological function.
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Objective To study the effect of ING5 gene on growth inhibition of gastric cancer. Methods ING5 expressing plasmid was transfect?ed into SGC?7901 cells. The cell viability was assessed by CCK?8,cell apoptosis and cycle was detected by flow cytometry analysis,the migration and invasion was evaluated by scratch and transwell,the expressions of mRNA and protein were determined by real?time quantitative PCR and West?ern blot respectively. Nude mice were implanted subcutaneously with SGC?7901 cells and tumor size and related protein were analyzed. Results After transfection of pEGFP?N1?ING5,the proliferation of gastric cancer SGC?7901 cells was significantly inhibited(P<0.05),the apoptosis was decreased(P<0.05),the percentage of G1 phase was increased and G2 phase was decreased(P<0.05),the ability of migration and invasion was reduced(P<0.05),the expression of NF?κB,PI3K,p?Akt1/2/3,Bcl?2,XIAP,β?catenin,c?myc mRNA,and protein levels were significantly in?creased(P<0.05),and the expression of Akt1/2/3、MMP9 mRNA and protein levels were significantly decreased(P<0.05). Conclusion The ING5gene inhibits the SGC?7901 cell proliferation,induces G1 arrest,inhibits the migration and invasion,and effectively regulates the related genes and proteins about cell proliferation,cell cycle and adhesion,but reduces apoptosis. ING5 can inhibit the evolution and development of gastric can?cer.
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Objective To establish model of the chicken embryo transplantation of human colon cancer cells ,and investigate the effect of Solanine、VEGF antibody and Solanine combined with VEGF antibody on human colon cancer cells induce tumor angio‐genesis and tumor proliferation .Methods The model of the chicken embryo transplantation of human colon cancer HT‐29 cells were divided into three experimental group and control group .We added to the chick embryo chorioallantoic membrane with Sola‐nine、VEGF antibody and Solanine+ VEGF antibody mixture ,PBS was added to the control group .Then we analysed picture through the stereomicroscope and IPP 6 .0 image analysis software ,using immunohistochemistry envision method to detect of CD34 antigen and ki‐67 antigen ,and observing effect of Solanine group ,VEGF antibody group ,Solanine+ VEGF antibody group and the effect on the tumor angiogenesis and tumor proliferation .Results The tumor angiogenesis ,CD34 antigen and ki‐67 antigen of Sola‐nine+VEGF antibody group were significantly better than those of VEGF antibody group and Solanine group(P<0 .01);VEGF antibody group had statistical significant difference with Solanine group(P<0 .01);the effect of other three groups were better than that of the control group(P<0 .01) .Conclusion Solanine、VEGF antibody and Solanine combined with VEGF antibody could in‐hibit tumor angiogenesis and tumor proliferation of human colon cancer cell line HT‐29 to induce .It provides a new way for anti‐an‐giogenes .
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Objective To explore the effects and molecular mechanisms of suberoylanilide hydroxamic acid (SAHA) on ovarian carcinoma. Methods (1)Two groups of ovarian carcinoma cell lines (SKOV3 and SKOV3/DDP, HO8910 and HO8910-PM) were exposed to SAHA (1, 3, 5 and 7μmol/L SAHA,group 1-group 4). CCK-8 method was employed to eval-uate the inhibitory effects of SAHA.(2)Ovarian cancer cell lines treated with SAHA (2 or 5μmol/L SAHA) were used as 1 and 2 groups. Flow cytometry was performed following staining with Annexin V-FITC and PI for cell cycle and apoptosis.(3) Reverse transcription polymerase chain reaction (RT-PCR) and Western blot assay were used to assess the mRNA and pro-tein expression levels of phenotypic correlation factor. Results (1)After 48 h of SAHA treatment,the OD value of SKOV3, SKOV3/DDP,and HO8910 showed a trend of gradually reduce (P<0.05).(2)The apoptotic rates were significantly higher in SAHA 1 and SAHA 2 groups than those of control group (P<0.05). Compared with control group, after 48 h of SAHA treat-ment,S phase and G2/M phase of SKOV3 and SKOV3/DDP cells increased;G0/G1 phase of HO8910 and HO8910-PM cells increased in SAHA 1 and 2 groups (P<0.05).(3)The expression levels of CyclinB1 and Cdc2 (p34) mRNA were significant-ly lower in SAHA 1 and 2 groups than those of control group,while the expression levels of Caspase-3,p21 and p53 mRNA expression were significantly higher in SAHA 1 and 2 groups than those of control group. Furthermore,the expression of Ac-Histone H3,Ac-Histone H4,p53 protein were markedly improved,and CyclinB1,Cdc2(p34) protein decreased in SAHA 1-4 groups. Conclusion SAHA may suppress cell growth, induce apoptosis and cause cycle arrest in ovarian carcinoma cells by promoting histone acetylation or modulating their phenotype-related proteins of Caspase-3, p53, CyclinB1 and Cdc2(p34).
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Objective to explore the role of Beclin 1 in gastric carcinogenesis and subsequent progression. Methods MkN28 cells were trans-fected with Beclin 1-expressing plasmid,and then the proliferation and cell cycle was measured by CCk-8 and PI staining. Beclin 1 expression was examined by immunohistochemistry and in situ hybridization on tissue microarrays containing gastric cancers,adjacent non-neoplastic mucosa,and metastatic lymph node. the correlation with the tumorgenesis,clinicopathological and prognostic parameters was analyzed. Results Beclin 1 overex-pression resulted in G2 arrest of MkN28 cells and reduced the proliferation. Beclin 1 mRNA was highly expressed in gastric cancer than matched mu-cosa by ISH(P < 0.05). Beclin 1 was highly expressed in male than female patients with gastric cancer(P < 0.05). the elder patients with gastric cancer had higher Beclin 1 expression than younger ones(P < 0.05). the diffuse-type carcinomas showed less Beclin 1 expression than intestinal and mixed type ones(P < 0.05). kaplan-Meier analysis indicated that Beclin 1 expression was positively correlated to favorable prognosis of the can-cer patients(P < 0.05). Conclusion Beclin 1 expression is closely linked to pathogenesis,metastasis and differentiation of gastric cancer. Beclin 1 might be employed to indicate the favorable prognosis of gastric cancer patients and regarded as a target of gene therapy.