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Objective To determine the association of one single nucleotide polymorphism loci G 398C in PRM2 with male in‐fertility in Chinese Han population .Methods A total of 386 infertile men were recruited as the observation group and 255 fertile men were recruited as the control group .Routine semen analysis as well as sperm functional parameters such as DNA integrity and nucleoprotein maturity rates were analyzed .Direct sequencing of G398C in PRM2 gene of infertile and fertile men was also conduc‐ted to evaluate the association of G398C SNP loci with male infertility .Results Statistical analysis showed that the frequencies of CC genotype of PRM2 G398C was significantly different between the infertile (11 .92% ) and fertile men (6 .67% ) and it was asso‐ciated with increased risk of male infertility (OR= 2 .002 ,95% CI= 1 .097 -3 .653 ,P< 0 .05) .Moreover ,it was discovered that sperm DNA integrity as well as nucleoprotein maturity rate of CC genotype were dramatically decreased compared with other geno ‐types (P<0 .05) ,which would probably lead to infertility .Conclusion Our results gave the first evidence that PRM 2 G398C poly‐morphism was associated with male infertility in Chinese Han population .
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Objective To analyse clinical application of hepatitis B virus core antibody(HBcAb)detected by using the chemilu-minescence microparticle immunoassay.Methods A total of 1 6 830 specimen with positive HBcAb detected by using the two pairs of semi-hepatitis test from January 2012 to November 2014 were collected,and divided into three groups according to the cut off in-dex(COI)of detection results of HBcAb,including group 1.0-0.05).The detec-tion rate of HBsAg(+)HBsAb(-)of group 9.0-<1 1.0 was significantly lower than that of the other two groups(P <0.05).A total of 304 specimen were HBsAg(-)HBsAb(-)and COI≥1 1,among them 64 specimen were HBV DNA postive and the posi-tive rate was 21.0%.Conclusion In the detection of HBcAb,COI≥1 1 and 1.0-<9.0 could be reference indicators for diagnosiing current and past HBV infection respectively,which should be combined with other laboratory indicators of HBV clinical data for comprehensive analysis.
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Objective: To investigate the effect of the gene interfering technology on fatty acid synthase (FAS) gene silencing for lipid contents in human hepatic cell line HepG2 and to study the lipid metabolism related gene expression in HepG2 cells. Methods: A total of 3 pairs of small interfering RNA (siRNA) targeting different sequences of FAS mRNA were synthesized as FAS-siRNA-1, FAS-siRNA-2 and FAS-siRNA-3, meanwhile, 2 controls were established as Blank control group, in which HepG2 cells were not treated, and Negative control group, in which HepG2 cells were transfected by non-effective siRNA. The mRNA, and protein expression levels of FAS in HepG2 cells were examined by real-time lfuorescence quantitative RCR and Western blot analysis to screen the most effective pair of siRNA for FAS gene silencing; and that speciifc siRNA was transtected to HepG2 cells for 48 hours to detect the intra-/extra-cellular TG, TC levels and the mRNA expression related to lipid metabolism in HepG2 cells. Results: The screening experiment indicated that FAS-siRNA-3 was most effective for FAS gene silencing. Compared with Blank control group, the mRNA and protein expressions in FAS-siRNA-3 transfected HepG2 cells (Transfected group)decreased to (52.33 ± 3.07) % and (51.57 ± 3.14) % respectively. Compared with Blank control group, Transfected group had the reduced intra-/extra-cellular TG levels and reduced extracellular TC level; while increased mRNA expression of hepatic lipase,P<0.0001 and decreased mRNA expression of TG transfer protein in HepG2 microsome,P<0.05. Conclusion: FAS gene silencing could signiifcantly decrease the intra-/extra- cellular TG level and extracellular TC level in HepG2 cells, those ifndings need to be conifrmed by furtherin vivo andin vitro studies.
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Objective To investigate the effect of fructose-1,6-diphosphate (FDP) pretreatment on myocardial connexin 43 (Cx43) in a rat model of acute myocardial ischemia.Methods Thirty-six male Sprague Dawley rats (aged 8-12 weeks and weighing 220-280 g) were randomly divided into three groups (n =12 each):sham operation group (group S),ischemia group (group Ⅰ) and FDP + ischemia group (group F).The animals were anesthetized with intraperitoneal injection of 10％ chloral hydrate 40 mg/100 g,then tracheostomized and mechanically ventilated.Acute myocardial ischemia was induced by occlusion of the left anterior descending coronary artery for 30 minutes.Myocardial ischemia was verified by elevation of the S-T segment on echocardiogram (EGG).In group F,FDP 100 mg/kg was injected intravenously 10 minutes before ischemia.The hearts were removed after 30 minutes of myocardial ischemia.The myocardial infarct size (IS) and area at risk (AAR) were measured and the IS/AAR ratio was calculated.The expression of myocardial Cx43 protein was determined by immunohistochemestry and analysis of mean optical density.Results The severities of arrhythmia were significantly higher in groups F and I than in group S,while lower in group F than in group Ⅰ (P＜ 0.05).The IS/AAR ratio was significantly lower in group F than in group Ⅰ.The myocardial Cx43 protein expression was down-regulated in group Ⅰ and group F as compared with group S,and was significantly lower in group Ⅰ than in group F.Conclusion FDP pretreatment can protect myocardium against acute ischemia by up-regulation of myocardial Cx43 expression.
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Objective To investigate the effect of sevoflurane anaesthesia on the cognitive function and phosphorylation of tau protein in hippocampal neurons in amyloid precursor protein (APP) transgenic mice.Methods Male APP gene mutation mice,weighing 18-22 g and aged 8-12 weeks,were used in this study.Forty-four APP positive mice were randomly divided into two groups:sevoflurane group (group AS,n =28) and control group (group AC,n =16).And other forty-four APP negative mice were randomly divided into two groups:sevoflurane group (group S,n =28) and control group (group C,n =16).The animals in groups S and AS inhaled 3％ sevoflurane for 4 hours.While in groups C and AC,the animals inhaled pure oxygen for 4 hours.Morris water maze was performed 24 hours after sevoflurane or pure oxygen inhalation.The phosphorylation of tau protein at Ser262 and Ser396 sites was detected by Western blotting on 1 day after pure oxygen inhalation in groups AC and C,and on 1,3 and 7 days after sevoflurane inhalation in groups AS and S.Results Compared with group C,the escape latency was significantly prolonged and the duration of staying at the original platform quadrant was shortened in groups S and AC,and the phosphorylation of tau protein at Ser262 site in group S and phosphorylation of tau protein at Ser262 and Ser396 sites in group AS were increased (P ＜ 0.05).Compared with group S,the escape latency was significantly prolonged,the duration of staying at the original platform quadrant was shortened,and the phosphorylation of tau protein at Ser262 and Ser396 sites was increased in group AS (P ＜ 0.05).Compared with group AC,the escape latency was significantly prolonged,the duration of staying at the original platform quadrant was shortened,and the phosphorylation of tau protein at Ser262 and Ser396 sites was increased in group AS (P＜0.05).Conclusion Sevoflurane anesthesia can aggravate the impairment of cognitive function in APP positive mice and the increase in the phosphorylation of tau protein at Ser262 and Ser396 sites is involved in the mechanism.
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Objective To evaluate the effects of sulforaphane preconditioning on cognitive dysfunction induced by sevoflurane anesthesia in aged rats.Methods Thirty Sprague-Dawley rats,aged 22 months,weighing 380-560 g,were randomly divided into 3 groups (n =10 each) using a random number table:control group,sevoflurane group (Sev group),and sulforaphane group (Sul group).In group Sul,sulforaphane25 mg/kg was administered by oral gavage once a day for 7 consecutive days,while the equal volume of distilled water was given instead of sulforaphane in Sev and C groups.Groups Sev and Sul inhaled 3％ sevoflurane in oxygen (2.5 L/min) and group C inhaled air (100 min/d) for 5 consecutive days starting from the end of gavage.At 24 h after sevoflurane inhalation,cognitive function was detected using Morris water maze and open field tests.The escape latency,frequency of crossing the original platform,the number of crossing the grid,the number of standing on the back legs and the time the animals spent in the central square were recorded.Results Compared with group C,the frequency of crossing the original platform,the number of crossing the grid,and the number of standing on the back legs were significantly reduced,the time the animals spent in the central square and escape latency on 1st day were prolonged and no significant changes were found in the escape latency on 2nd-4th days in group Sev.Compared with group Sev,the frequency of crossing the original platform,the number of crossing the grid,and the number of standing on the back legs were significantly increased,and the time the animals spent in the central square and escape latency on 1st day were shortened.Conclusion Sulforaphane preconditioning can improve the cognitive dysfunction induced by sevoflurane anesthesia in aged rats.
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Objective To investigate the effect of sevoflurane anesthesia on the cognitive function and phosphorylation of tau in hippocampal neurons in amyloid precursor protein (APP) transgenic mice.Methods Male APP gene mutation mice,weighing 18-22 g,aged 8-12 weeks,were used in the study.Forty-four APP positive mice were randomly divided into 2 groups (n =10 each):sevoflurane group (group AS,n =28) and control group (group AC,n =16).Forty-four APP negative mice were randomly divided into 2 groups:sevoflurane group (group S,n =28) and control group (group C,n =16).Animals in groups S and AS inhaled 3％ sevoflurane for 4 h.While in groups C and AC,animals inhaled pure oxygen for 4 h.Morris water maze was performed 24 h after sevoflurane or pure oxygen inhalation.The phosphorylation of tau at Ser262 and Ser396 was detected by Western blot on 1 day after pure oxygen inhalation (T1) in groups AC and C,and on 1,3 and 7 days after sevoflurane inhalation in groups AS and S.Results Compared with group C,the escape latency was significantly prolonged and the duration of staying at the original platform quadrant was shortened in groups S and AC,and the phosphorylation of tau at Ser262 in group S and phosphorylation of tau at Ser262 and Ser396 in group AS were increased (P ＜0.05).Compared with group S,the escape latency was significantly prolonged,the duration of staying at the original platform quadrant was shortened and the phosphorylation of tau at Ser262 and Ser396 was increased in group AS (P ＜ 0.05).Compared with group AC,the escape latency was significantly prolonged,the duration of staying at the original platform quadrant was shortened and the phosphorylation of tau at Ser262 and Ser396 was increased in group AS (P ＜ 0.05).Conclusion Sevoflurane anesthesia can aggravate the impairment of cognitive function in APP positive mice and the increase in the phosphorylation of tau at Ser262 and Ser396 is involved in the mechanism.
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ObjectiveTo investigate the effect of sevoflurane preconditioning on heme oxygenase-1 (HO-1) expression in lung tissues during one lung ventilation (OLV) in rats.MethodsTwenty-four male SD rats weighing 220-250 g were randomly divided into 4 groups ( n =6 each):control group (group C) ; two lung ventilation group (group T); OLV group (group O) and sevoflurane preconditioning+ OLV group (group SO).The animals were anesthetized with intraperitoneal pentobarbital sodium 40 mg/kg.In group T,the animals were tracheal intubated and bilateral lungs were ventilated for 1 h (VT 10 ml/kg,RR 60 bpt/min,I∶E 1∶2) and PETCO2 was maintained at 35-50 mm Hg.In groups O and SO,the animals were tracheal intubated and OLV was performed for 1 h (VT 5 ml/kg,RR 80 bpt/min,I:E 1:2) and PETCO2 was maintained at 35-50 mm Hg.In group SO,2.4％sevoflurane was inhaled for 30 min before OLV and then washed out by inhalation of oxygen for 15 min.The left lung tissues were removed in groups C and T,and the bilateral lung tissues were removed in groups.O and SO for microscopic examination and determination of wet/dry lung weight ratio (W/D ratio) and expression of HO-1 (by Western blot).Results Compared with group C,W/D ratio was significantly increased and the expression of HO-1 was up-regulated in left lung tissues in group T and in bilateral lung tissues in groups O and SO ( P ＜0.05).Compared with group T,W/D ratio was significantly increased and the expression of HO-1 was up-regulated in bilateral lung tissues in group O and in right lung tissues in group SO ( P ＜ 0.05).Compared with group O,W/D ratio was significantly decreased,the expression of HO-1 was up-regulated (P ＜ 0.05) and the pathological changes were reduced in bilateral lung tissues in group SO.ConclusionThe mechanism by which sevoflurane preconditioning reduces OLV-induced lung injury is related to up-regulation of HO-1 expression.
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Objective To investigate the effect of fructose-1,6-diphosphate (FDP) pretreatment on myocardial connexin43 (Cx43) in a rat model of acute myocardial ischemia.Methods Thirty-six male 8-12 week old SD rata weighing 220-280 g were randomly divided into 3 groups (n=12 each):group Ⅰ sham operation (group S);group Ⅱ ischemia(group Ⅰ)and group Ⅲ FDP+ischemia(group F).The animals were anesthetized with intraperitoneal 10%chloral hydrate 40 mg/100 g,tracheostomized and mechanically ventilated.Acute myocardial ischemia was induced by occlusion of left anterior descending coronary artery for 30 min.Myocardial ischemia was;verified by elevation of S-T segment on ECG.In group F FDP 100 mg/kg was injected iv at 10 min before ischemia.Arrhythmia was recorded within 30 min after occlusion and the severity of arrbythmia was aggesged.The hearts were removed after 30 min myocardial ischemia.The Left ventricle area (LVA),myocardial infarct area (IA) and area at risk (AAR) were measured and AAR/LVA and IA/AAR ratios were calculated.The expression of myocardial Cx43 protein was determined by immuno-histochemestry and analysis of mean optical density.Results The severity of arrhythmia was significantly higher in group F and I than in gropu S.while lower in group F than in group I(P<0.05).The IA,IA/AAR ratio was significantly lower in group F than in group I.The myocardial Cx43 protein expression was down-regulated in group I and F as compared with group S.and was significantly lower in group I than in group F.Conclusion FDP pretreatment can protect myocardium against acute ischemia by up-regulation of myocardial Cx43 expression.
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Objective To compare the effects of flurbiprofen axetil and fentanyl of postoperative analgesia on T-lymphocytes subtypes in patients undergoing esophagectomy.Methods Forty patients undergoing esophagectomy were randomly divided into two groups(20 cases each):group C (group fentanyl) was given fentanyl 20μg/kg plus tropisetron 5 mg diluted to 100 ml via PCIA after surgery,group F (group flurbiprofen-fentanyl) was administrated flurbiprofen axetil 50 mg at 30 min before the end of surgery,fentanyl 10μg/kg and flurbiprofen axetil 100 mg plus tropisetron 5 mg diluted 100 ml was administrated via PCIA after surgery.The PCIA rate was 2 ml/h,bolus 0.5ml,lock time 15 min.The VAS score was recorded at 12,24,48 h after surgery.Blood samples 2 ml were obtained from peripheral vein for determination of CD3+,CD4+,CD8+ and CD4+/CD8+ at 30 min before surgery(T0),24 h(T1)and 72 h(T2)after surgery.Results Patients in two groups did not show any significant difference in the VAS scores(P＞0.05).At T1 CD3+,CD4+ T-lymphoeytes were significantly lower than those at T0 in two groups(P＜0.05).At T2,CD3+,CD4+ T-lymphocytes in group F were significantly higher than those in group C(P＜0.05).In group C,CD3+,CD4+ T-lymphocytes at T2 were significantly decreased than those at T0(P＜0.05).CD8+ T-lymphocytes Was no significantly changed in two groups and at each time point determined (P＞0.05).Conclusion Postoperative analgesia by using flurbiprofen axetil and fentanyl can diminish the using dose of postoperative opioid drug,and it can'improve patient's cellular immune function.
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Objective To investigate the effect of general anesthesia with sevoflurane or intravenous propofol on anesthesia recovery period in obstructive jaundice patients. Methods Thirty ASA Ⅰ or Ⅱ and Child A obstructive jaundice patients were randomly divided into two equal groups (n=15 each). The patients in group S received inhalation anesthesia with sevoflurane and those in group P intravenous anesthesia with propofol during operation for obstructive jaundice. The patients were premedicated with intramuscular phenobarbital 100mg and atropine 0.5mg, anesthesia was induced with midazolam 0.05mg/kg, atracurium 0.5mg/kg, propofol 1.5-2.5mg/kg and fentanyl 4μg/kg. Maintained with TCI of propofol (target plasmaconcentration was set at 3.5mg/L) or sevoflurane inhalation (end-tidal sevoflurane concentration was 2%-3%) and intermittent i. v. boluses of fentanyl. EGG, HR, MAP, SpO<,2> and end-tidal sevoflurane concentration were continuously monitored during operation. Duration of anesthesia, the volume of infusion and fentanyl were recorded, awaking time, extubation and regained consciousness after operation were recorded. Results There were no significant differences between the two groups in average age, sex, body-weight, duration of anesthesia, the parameters of MAP and HR (P>0.05). The awaking time was (7.9±1.5) minutes in group S and (26.1±8.8) minutes in group P. The extubation time was (8.5±2.5) minutes in group S and (27.8±11.2) minutes in group P. The regained consciousness time was (13.1±4.4) minutes in group S and (33.7±12.5) minutes in group P. The incidence of lethargy, fidget were higher in group P than those in group S. Conclusion Both sevoflurane and propofol can provide satisfactory anesthesia for the operation of obstructive jaundice, but the recovery of influence caused by sevoflurane is faster and more steady than that caused by propofol.
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The inhibition of metastatic progression of Somatostatin receptor type 2 (SSTR2) gene transfection mediated by adenovirus in human pancreatic carcinoma cells and the mechanisms involved in this effect were studied. The full-length human SSTR2 cDNA was introduced into the pancreatic cancer cell line BXPC-3 by adenovirus-mediated transfection. Stable expression of mRNAs and protein of SSTR2 was detected by RT-PCR and Western-blot. The Matrigel-coated Transwell was used to detect the migratory and invasive ability of SSTR2-expressing cells, Adv-GFP control cells and mock control cells. Furthermore, the expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) was detected byRT-PCR in these cells. The stable expression of SSTR2 was detected in BXPC-3 transfected by Adv-GFP-SSTR2. A dramatic decrease of BXPC-3 expressing sst2 cells migrating through a Matrigel-coated filter was observed, as compared with Adv-GFP control and mock control cells (P＜0.01). Moreover, the expression of MMP-2 mRNA was significantly reduced in the SSTR2-expressing cells and conversely the expression of TIMP-2 mRNA was significantly increased in the SSTR2-expressing cells when compared with the Adv-GFP control and mock control (P＜0.01). The expression of reintroduced human SSTR2 gene in BXPC-3 cells by Adv-GFP-SSTR2 had the anti-migratory and anti-invasive effects, and the mechanisms involved in this effect may be due to the down-regulated expression of MMP-2 and up-regulated expression of TIMP-2.
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Extracellular matrix (ECM) degradation is an essential step that allows tumor cells to penetrate a tissue barrier and become metastatic. Heparanase (HPSE) is an endoglycosidase that specifically degrades heparin sulfate proteoglycans (HSPG), a chief component of ECM. HPSE is not expressed in normal epithelial cells but can be detected in a variety of human carcinomas including pancreatic cancer. In the present study, human pancreatic cancer cell line Panc-1 was transfected with HPSE antisense oligodeoxynucleotide (AS-ODN) in vitro, then the inhibitory effect of ASODN on HPSE gene expression and invasive ability of Panc-1 cells in vitro was examined. The HPSE mRNA and protein expression of Panc-1 cells transfected with AS-ODN was significantly inhibited. However, there were no marked inhibitory effects in Panc-1 cells treated with nonsense oligodeoxynucleotide (NS-ODN). Moreover, a modified Boyden chamber assay demonstrated that transfection with HPSE AS-ODN significantly inhibited invasive potential of Panc-1 cells in vitro after AS-ODN transfection. This suggests that HPSE AS-ODN may contribute to the inhibition of HPSE mRNA and protein expression, and results in a decrease of the invasive ability of Panc-1 in vitro.
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Objective To explore the expression and clinical significance of EphA2 and E-cadherin in (pancreatic) cancer tissue and cells.Methods The expression of EphA2 and E-cadherin in 56 pancreatic(carcinomas) and 23 adjacent noncancer tissues were studied by immunohistochemistry,and their relationship to clinicopathological characteristics were analyzed.RT-PCR was performed to explore the expression of EphA2,BXPC-3,PC-3 and PANC-1 in pancreatic carcinoma cell lines.Results In pancreatic carcinomas tissuse showed increased EphA2 expression and reduced E-cad expression which compared with adjacent noncancer tissues.The expression level of EphA2 had a significant positive relationship with tumor differentiation degree,lymphatic metastasis and clinical stage.However,the expression level of E-cadherin had negative relationship with both the tumor clinical stage and lymphatic invasion.Furthermore,a significant negative relationship(between) the expression of EphA2 and E-cadherin was observed.The expressions of EphA2 were higher in high-invasive cell lines PXPC-3 and Panc-1 than in low-invasive cell line PC-3.Conclusions The(expression) and/or abnormal function of EphA2 and E-cadherin may together be involved in the development and progression of pancreatic cancer;the combined measurement of these two proteins may be useful for(determination) of metastatic potency of pancreatic carcinoma.
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Objective To investigate the anti-migratory and anti-invasive effect of somatostatin receptor type 2(SSTR2) gene transfection mediated by adenovirus in human pancreatic carcinoma cell and the mechanisms involved in this effect.Methods The full length human SSTR2 cDNA was introduced into pancreatic cancer cell line BXPC-3 by adenovirus-mediated transfection,and stable expression of RNA and protein of SSTR2 were detected by RT-PCR and Westen-blot.The Matrigel coated Transwell was used to detect the migratory and invasive ability of SSTR2expressing cells,Adv-GFP control cells and mock control cells.Furthermore,the expressions of matrix metalloproteinase-2(MMP-2) and tissue inhibitor of metalloproteinase-2(TIMP-2) were detected by RT-PCR method in these cells.Results The stable expression of SSTR2 was detected in BXPC-3 cells transfected by Adv-GFP-SSTR2.A dramatic decrease of BXPC-3 expressing SSTR2 cell(migrated) through a Matrigel-coated filter was observed,as compared with Adv-GFP control cells and mock control cells(P
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Objective To investigate the effect of integrin-?1 antisense oligodeoxynucleotide(ASODN) on(human) pancreatic cancinoma transplanted subcutaneously in nude mice.Methods The models of human(pancreatic) cancinoma transplanted subcutaneously were established in nude mice,then divided randomly into 3 groups and different treatment was given respectively(control group,random oligodeoxynucleotide group and ASODN group).After treatment,the weight of nude mice and tumor volume were observed,and the tumor growth inhibitory rate and the tumor response rate were calculated.The expressions of integrin-?1 mRNA and protein in tumor tissue were determined by RT-PCR and Western-blot.Results The tumor growth inhibitory rate in the random oligodexynucleotide group and the ASODN group was 4.75% and 72.70%,respectively.The tumor decrease rate of the ASODN group was 10.91%.The expression level of integrin-?1 mRNA and protein was decreased in the ASODN group compared with other 2 control groups. Conclusions Our findings suggest that integrin-?1 antisense oligodeoxynucleotides result in marked inhibition of human pancreatic(cancinoma) growth in nude mice.It may be a novel treatment approach for human pancreatic carcinoma.
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Objective To observe the effect of antisense hypoxia inducible factor-1?(HIF-1?) on (chemosensitivity) of human pancreatic cancer cell line BxPC-3 under hypoxia. Methods BxPC-3 cells were divided into 3 groups:(1)BxPC-3 cells were non-transfected with antisense HIF-1? plasmid and exposed to 0.5% O_2 for 4hr(hypoxia control);(2)normoxic BxPC-3 cells were non-transfected with antisense(HIF-1?) plasmid(normoxia control);(3)BxPC-3 cells were transfected with antisense HIF-1? plasmid and exposed to 0.5% O_2 for 4hr(experimental group).Expression of HIF-1? and survivin was detected by RT-PCR and Western Blot.Growth inhibition rates and apoptosis rates of BxPC-3 cells under different(dosages) of chemotherapeutic agents(5-fluorouracil,doxorubicin and gemcitabine) were measured by MTT(colorimetric) assay and flow cytometry (FCM).Results Expression of HIF-1? was obviously down-regulated and at the same time susvivin expression was markedly down-regulated in experimental group(P