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Objective@#To construct 3β-HSD gene shRNA lentivirus interference vecto, then transfect into human MCF-7 cells, and construct cell line with 3β-HSD gene silencing, finally to study the effects of 3β-HSD on apoptosis induced by di- (2-ethylhexyl) phthalate (DEHP) .@*Methods@#According to the mRNA sequence of 3β-HSD gene provided by GenBank, three interference sequences were designed and connected to PLVX-shRNA2-puro after annealing. The recombinant lentivirus vector was transfected into 293FT cells, the virus supernatants were collected and infected with MCF-7 cells. After puromycin screening, MCF-7 cells with 3β-HSD gene silencing were constructed. The cells with 3β-HSD gene silencing were identified by real-time quantitative PCR and western blot. Then the 3β-HSD gene silencing cells and MCF-7 cells were treated at various doses of DEHP for 24 hours to detect the gene expression and protein expression of apoptosis genes including Bax, Caspase-3 and Caspase-8.@*Results@#The interference sequence of 3β-HSD gene inserted into lentivirus vector PLVX-shRNA2-puro is consistent with the designed sequence. 3β-HSD gene expression level in MCF-7 cells with 3β-HSD gene silencing was 77% lower than than that of control MCF-7 cells. 3β-HSD protein level in MCF-7 cells with 3β-HSD gene silencing was 74% lower than that of control MCF-7 cells. After DEHP treatment in MCF-7 cells with 3β-HSD gene silencing and control MCF-7 cells, qRT-PCR results showed that Bax gene expression levels increased by 28%-54%, Caspase-3 gene increased by 13%-49%, Caspase-8 gene increased by 21%-70% in MCF-7 cells when compared with the control group. Additionally, in the 3β-HSD gene silencing cells, Bax gene expression level decreased by 11%-28%, Caspase-3 gene expression decreased by 12%-23%, Caspase-8 gene expression decreased by 11%-34%, compared with the same treatment group of MCF-7 cells. Western blot results showed that Bax protein expression level increased by 28%-61%, Caspase-3 protein expression level increased by 40%-48%, Caspase-8 protein increased by 31%-84% in MCF-7 cells when compared with the control group. In 3β-HSD gene silencing cells, Bax protein expression level increased by 11%-27%, Caspase-3 protein increased by 21%-40%, Caspase-8 protein increased by 12%-25%, compared with the same treatment group of MCF-7 cells.@*Conclusion@#The stable 3β-HSD gene silencing cell line are successfully constructed in this study. DEHP can induce increased expression of apoptotic gene and protein. Silencing of 3β-HSD gene can inhibit the activation of apoptotic gene by DEHP in a certain degree.
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Objective To evaluate the safety of Propofol combined with Fentanyl in intravenous anesthesia for endoscopic retrograde cholangiopancreatography(ERCP)in elderly patients.Methods The clinical data of 63 elderly patients aged ≥80 years with common bile duct stones who underwent ERCP in digestive endoscopy center of our hospital were retrospectively analyzed.All the patients were divided into Propofol combined with Fentanyl anesthesia group(group A,n =41)and midazolam combined with Fentanyl anesthesia group(group B,n=22).The changes of blood pressure,heart rate and blood oxygen saturation(SpO2),and postoperative complications were compared between the two groups.Results The incidence of hypotension was higher in Propofol combined with Fentanyl anesthesia group than in midazolam combined with Fentanyl anesthesia group(24.4 ％ vs.0.0％,x2=4.683,P =0.030),but the incidence of SpO2 ＜ 90 ％ in midazolam combined with fentanyl anesthesia group was significantly increased(22.7％ vs.0.0％,x2 =7.250,P =0.007).There was no statistically significant difference in the incidence of anesthesia-related complications (39.0％ vs.36.4％,x2 =0.043,P =0.836) and postoperative complications (4.9 ％ vs.4.5 ％,x2 =0.003,P =0.953) between the two groups.Conclusions For elderly patients with common bile duct stones,intravenous anesthesia with Propofol and Fentanyl during ERCP is safe,and blood pressure changes need to be closely monitored.
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Objective@#To study the oxidative damage of di- (2-ethylhexyl) phthalate (DEHP) on MCF-7 cells, and to investigate the effects of 3β-hydroxysteroid dehydrogenase (3β-HSD) gene silence or overexpression on DEHP-induced oxidative damage.@*Methods@#MCF-7 cells, 3β-HSD gene silencing cells and 3β-HSD gene overexpression cells were treated with different doses of DEHP (0,0.05,0.1,0.2,0.4,0.8 mmol/L) for 24h, then intracellular oxidative damage index such as MDA, SOD, GSH, GSH-PX were detected, DNA repair gene hOGG1, hMTH1 mRNA expression were tested by Q-PCR, hOGG1, hMTH1 protein expression were detected by western blot.@*Results@#After MCF-7 cells were treated by DEHP, MDA levels increased; SOD activity, GSH content, GSH-PX activity decreased, hOGG1 and hMTH1 mRNA expression levels increased, hOGG1 and hMTH1 protein expression levels increased, the differences were statistically significant when compared with control (P<0.05 or P<0.01) . In 3β-HSD gene silencing cells which were treated by DEHP, when compared with the same dose group of MCF-7 cells, MDA content increased, SOD activity, GSH content, GSH-PX activity decreased, hOGG1 and hMTH1 mRNA expression levels decreased, hOGG1 and hMTH1 protein expression levels decreased, the difference were statistically significant (P<0.05 or P<0.01) . In 3β-HSD gene overexpression cells which were treated by DEHP, when compared with the same dose group of MCF-7 cells, MDA content decreased; SOD activity, GSH content, GSH-PX activity increased, of hOGG1 and hMTH1 mRNA expression levels increased, hOGG1 and hMTH1 protein expression levels increased, the difference were statistically significant (P<0.05 or P<0.01) .@*Conclusion@#DEHP could cause oxidative damage in MCF-7 cells, induce the changes of related genes and proteins, 3β-HSD plays an antioxidant role in the process of DEHP ox-idative damage.
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Objective To discuss the clinical aesthetic effect of vermilion repair on cleft lips after using double-line reconstruction.Methods 68 children with cleft lip were repaired according to the principle of double-line reconstruction,and two contents including 7 projects,such as shapes of upper lips,vermilion borders of lips,vermilion shapes and lips,etc,were evaluated comprehensively within one month after operation for assessing the postoperative effect.Results 68 cases were followed up,with the follow-up rate of 100％,and 62 cases (91.2％) were satisfied with the vermilion repair.Six cases (8.8％) had a common effect of vermilion shapes after repair.In the 7 contents evaluated,satisfactory results were achieved in the projects except the relatively poor symmetry vermilion borders of lips.Conclusions The double-line reconstruction helps to improve the aesthetic effect of vermilion repair on cleft lips.
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<p><b>OBJECTIVE</b>To investigate the effects of trichloroethylene (TCE) toxicity on the normal human liver cells (L02 cells) and hepatocytes with CYP2E1 gene overexpression which was constructed through molecular cloning technology in our laboratory, then to explore the roles of CYP2E1 gene in TCE toxicity.</p><p><b>METHODS</b>L02 cells and hepatocytes with CYP2E1 overexpression were treated with various doses of TCE (0,0.25, 0.5, 1.0, 2.0, 4.0 mmol/L) for 12h, the expression of apoptosis genes (Bcl-2, Caspase-3, Caspase-8, Caspase-9) and oncogenes (c-fos, c-myc, k-ras, p53) were determined by real-time fluorescent PCR.</p><p><b>RESULTS</b>Bcl-2 mRNA expression levels increased significantly in normal liver cells and CYP2E1-overexpressing cells after TCE treatment, Bcl-2 levels were 20%∼50%higher in CYP2E1-overexpressing cells than in L02 liver cells at doses of 0.25∼2.0 mmol/L TCE. Caspase-3, Caspase-8 and caspase-9 mRNA expression increased by 30%∼600% in CYP2E1-overexpressing cells at doses of 0.5∼4.0 mmol/L TCE when compared with L02 cells (P < 0.01). Additionally, c-fos, k-ras and c-myc mRNA expression levels were 25%∼120% higher in CYP2E1-overexpressing cells than in L02 cells (P < 0.01), p53 mRNA expression levels were lower 10%∼50% in CYP2E1-overexpressing cells than in L02 cells (P < 0.05 or P < 0.01).</p><p><b>CONCLUSIONS</b>There were significant differences for apoptosis gene and oncogene expression levels between normal liver cells and CYP2E1-overexpressing cells after they were treated with TCE, these findings indicated that CYP2E1 might play an important role in TCE metabolism in vivo.</p>
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Humanos , Proteínas Reguladoras de la Apoptosis , Caspasa 3 , Caspasa 8 , Caspasa 9 , Citocromo P-450 CYP2E1 , Genética , Expresión Génica , Hepatocitos , Hígado , Proto-Oncogenes , Genética , ARN Mensajero , Tricloroetileno , ToxicidadRESUMEN
ObjectiveTo investigate the clinical value of serum cystatin C(CysC) and urinary microalbumin(MA) in the diagnosis of early renal function injury in patients with essential hypertension.MethodsOne hundred and twenty patients with essential hypertension(hypertension group) were divided into three groups by the results of blood pressure:grade 1 with 48 cases,grade 2 with 47 cases,grade 3 with 25 cases.Thirty healthy subjects were selected as control group.Serum CysC,urinary MA and serum creatinine (Cr) were detected in all subjects.ResultsThe serum CysC and urinary MA in grade 1,2 and 3 hypertension group [(1.57 ±0.48),(2.12 ±0.72),(2.91 ± 1.09) mg/L and(18.12 ±5.43),(29.01 ±8.07),(46.06 ± 13.21 ) mg/L] were obviously higher than those in control group [ (0.71 ± 0.23 ),(9.35 ±5.17)mg/L](P＜ 0.05).The serum Cr had no significant difference between grade 1,2,3 hypertension group and control group (P ＞ 0.05 ).Serum CysC was positively correlated with urinary MA in hypertension group (r =0.613,P ＜ 0.05),serum CysC and urinary MA were both negatively correlated with estimated glomenlar filtration rate (eGFR)(r=-0.635,-0.563,P＜0.05).ConclusionsSerum CysC and urinary MA are sensitive indexes of early renal function injury in patients with hypertension.The combined determination of serum CysC and urinary MA can improve the detection rate of early renal function injury.
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Objective To observe the effect of chewing gum on EGG,blood motilin and anus exhausting after laparoscopic abdominal surgery. Methods 200 patients after laparoscopic cholecystectomy were randomly arranged into chewing gum group (Group G): to start chewing gum for 30 min at breakfast time on first day after operation, and also at lunch and dinner time, and stopped until the first anus exhausting, and the control group(Group C): no chewing gum. The time between endpoint of operation to first anus exhausting was recorded; blood motilin and electrogastrography (EGG)on the morning of 1,2,3 days postoperation after chewing gum was detected. Results The time between endpoint of operation to first anus exhausting was not significantly different between two groups, compared with the control group, blood motilin significantly improved on the first day and the second day,but there was no difference between two groups on 3rd day; EGG was no different in all three days between two groups. Conclusions Chewing gum doesn't improve bowel function after laparoscopic abdominal surgery.
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Objective To provide an ideal tissular flap for the soft tissue defects of the oral commissure. Methods The observation and measurement of the source of blood supply and nerve innervation to the great/second toe web was carried out on 40 sides of adult specimens lower limbs perfused with red latex in artery. Results The first dorsal metatarsal artery which originated from the dorsal pedal artery was the chief artery to this region. Its external diameter at the beginning was (l. 6 ? 0. 3) mm and was (4. 6?0. 6) cm in length. Sensory nerve of the flap is deep peroneal nerve Conclusion The great/second toe web flap was an ideal tissue defects of the oral commissure.
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Objective To explore the relationship between dopaminergic axon terminal and GABAergic neurons and explore the neuroanatomic mechanism of their effects in schizophrenia. Methods The co-location and the association of dopaminergic axon terminal and GABAergic neurons in the basolateral nucleus (BL) of rat amygdala were examined by using double labeling immunoelectron microscopic techniques. Dopaminergic axon terminal and GABAergic neurons were labeled with the antidopamine (anti-DA) and the anti-glutamic acid decarboxylase (anti-GAD) antibodies respectively. Results 43% of the DA-input synapses was observed to relate directly or indirectly to GAD-immunoreactive(IR) dendritic structures(DA/GAD), which includes single (38%), convergent (30%), serial (20%), and axoaxonic (12%) contact types.And 57% of the DA-input synapses was found to associate with unlabeled elements (DA/UE), which includes unlabeled perikarya (10%), dendrites (82%) and axons (8%). All of the synapses that show DA-IR terminals profiles were found to be symmetric (inhibitory) synapses.Conclusion These results suggest that the dopaminergic system in the BL of rat amygdala controls the mediations of the GABAergic interneurons via symmetric synapses. In addition, the dopaminergic axon terminal associates with the glutamatergic projection neurons and exerts influence on its activity.