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Background@#Tuberculosis is a leading cause of morbidity and mortality in humans worldwide. There is an urgent need for new and effective drugs to treat tuberculosis and shorten the duration of tuberculosis therapy. 1, 25-dihydroxy vitamin D3 (1,25 (OH)2D3) has been reported to have a synergistic effect with pyrazinamide (PZA) in killing tubercle bacilli in vitro. The addition of 1,25 (OH)2D3 to standard tuberculosis treatment should benefit patients if the adjunctive drug has a synergistic effect in vivo. Thus, in this study, calcitriol (bioactive 1,25 (OH)2D3) was administered to mice undergoing treatment for Mycobacterium tuberculosis (M.tb) infection with PZA, a first-line anti-tuberculosis drug, to determine whether vitamin D3 enhances the therapeutic effect.@*Methods@#C57BL/6 female mice were infected with the M.tb H37Rv strain through aerosol exposure. Calcitriol and PZA, either alone or in combination, were orally administered to the M.tb infected mice. The effect of calcitriol on PZA activity was determined by evaluating the bacterial burden and analyzing the histopathological lesions in the lungs and spleen. To investigate the expression of inflammatory cytokines and anti-microbial peptide genes, we determined the transcriptional levels of interferon-γ (IFN-γ), interleukin-4 (IL-4), mouse β-defensin-2 (mBD2), and cathelicidin LL-37 through real-time quantitative polymerase chain reaction. The protein levels of IFN-γ were detected by enzyme-linked immunosorbent assay. Differences between groups were analyzed with independent samples t-test or one-way analysis of variance.@*Results@#Calcitriol alone had little effect on tuberculosis infection, whereas PZA, compared with saline control treatment, decreased the bacterial burden (spleens: PZA vs. saline, 4.82 ± 0.22 vs. 5.22 ± 0.40 Log10 colony-forming units [CFU]/gram, t = 2.13, P < 0.05; lungs: PZA vs. saline, 5.55 ± 0.15 vs. 6.83 ± 0.46 Log10 CFU/gram, t = 6.56, P < 0.01) and pathological lesions in the lungs. Simultaneous administration of calcitriol with PZA, compared with PZA alone, decreased the bacterial load (spleen: calcitriol + PZA vs. PZA, 4.37 ± 0.13 vs. 4.82 ± 0.22 Log10 CFU/gram, t = 4.36, P < 0.01; lung: calcitriol + PZA vs. PZA, 5.03 ± 0.32 vs. 5.55 ± 0.15 Log10 CFU/gram, t = 3.58, P < 0.01) and attenuated the lung lesions (gross pathological score: calcitriol + PZA vs. PZA, 3.25 ± 0.50 vs. 2.50 ± 0.58, t = 1.96, P < 0.05; affected area of total lung area: calcitriol + PZA vs. PZA, 30.75% ± 6.50% vs. 21.55% ± 2.99%, t = 2.66, P < 0.05). Further studies demonstrated calcitriol significantly increased the expression of anti-inflammatory cytokine IL-4 but suppressed production of the pro-inflammatory cytokine IFN-γ (IL-4: calcitriol vs. saline, 5.69 ± 0.50 vs. 2.80 ± 0.56 fold of control, t= 6.74, P < 0.01; IFN-γ: calcitriol vs. saline, 1.36 ± 0.11 vs. 4.13 ± 0.83 fold of control, t= 5.77, P < 0.01). In addition, calcitriol alone or in combination with PZA significantly enhanced the transcriptional level of anti-microbial peptides (cathelicidin LL-37: calcitriol vs. saline, 10.59 ± 1.03 vs. 2.80 ± 0.90 fold of control, t = 9.85, P < 0.01; mBD2: calcitriol vs. saline, 7.92 ± 0.62 vs. 1.79 ± 0.45 fold of control, t = 13.82, P < 0.01), whereas PZA exerted a negative effect on anti-microbial peptide gene expression.@*Conclusions@#Calcitriol as adjunctive treatment can result in beneficial treatment outcomes in M.tb infection by suppressing the inflammatory response and up-regulating the expression of anti-microbial peptides. These results indicate the feasibility of using calcitriol adjunctively with standard chemotherapy for the treatment of M.tb infection.
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BACKGROUND@#Tuberculosis is a leading cause of morbidity and mortality in humans worldwide. There is an urgent need for new and effective drugs to treat tuberculosis and shorten the duration of tuberculosis therapy. 1, 25-dihydroxy vitamin D3 (1,25 (OH)2D3) has been reported to have a synergistic effect with pyrazinamide (PZA) in killing tubercle bacilli in vitro. The addition of 1,25 (OH)2D3 to standard tuberculosis treatment should benefit patients if the adjunctive drug has a synergistic effect in vivo. Thus, in this study, calcitriol (bioactive 1,25 (OH)2D3) was administered to mice undergoing treatment for Mycobacterium tuberculosis (M.tb) infection with PZA, a first-line anti-tuberculosis drug, to determine whether vitamin D3 enhances the therapeutic effect.@*METHODS@#C57BL/6 female mice were infected with the M.tb H37Rv strain through aerosol exposure. Calcitriol and PZA, either alone or in combination, were orally administered to the M.tb infected mice. The effect of calcitriol on PZA activity was determined by evaluating the bacterial burden and analyzing the histopathological lesions in the lungs and spleen. To investigate the expression of inflammatory cytokines and anti-microbial peptide genes, we determined the transcriptional levels of interferon-γ (IFN-γ), interleukin-4 (IL-4), mouse β-defensin-2 (mBD2), and cathelicidin LL-37 through real-time quantitative polymerase chain reaction. The protein levels of IFN-γ were detected by enzyme-linked immunosorbent assay. Differences between groups were analyzed with independent samples t-test or one-way analysis of variance.@*RESULTS@#Calcitriol alone had little effect on tuberculosis infection, whereas PZA, compared with saline control treatment, decreased the bacterial burden (spleens: PZA vs. saline, 4.82 ± 0.22 vs. 5.22 ± 0.40 Log10 colony-forming units [CFU]/gram, t = 2.13, P < 0.05; lungs: PZA vs. saline, 5.55 ± 0.15 vs. 6.83 ± 0.46 Log10 CFU/gram, t = 6.56, P < 0.01) and pathological lesions in the lungs. Simultaneous administration of calcitriol with PZA, compared with PZA alone, decreased the bacterial load (spleen: calcitriol + PZA vs. PZA, 4.37 ± 0.13 vs. 4.82 ± 0.22 Log10 CFU/gram, t = 4.36, P < 0.01; lung: calcitriol + PZA vs. PZA, 5.03 ± 0.32 vs. 5.55 ± 0.15 Log10 CFU/gram, t = 3.58, P < 0.01) and attenuated the lung lesions (gross pathological score: calcitriol + PZA vs. PZA, 3.25 ± 0.50 vs. 2.50 ± 0.58, t = 1.96, P < 0.05; affected area of total lung area: calcitriol + PZA vs. PZA, 30.75% ± 6.50% vs. 21.55% ± 2.99%, t = 2.66, P < 0.05). Further studies demonstrated calcitriol significantly increased the expression of anti-inflammatory cytokine IL-4 but suppressed production of the pro-inflammatory cytokine IFN-γ (IL-4: calcitriol vs. saline, 5.69 ± 0.50 vs. 2.80 ± 0.56 fold of control, t = 6.74, P < 0.01; IFN-γ: calcitriol vs. saline, 1.36 ± 0.11 vs. 4.13 ± 0.83 fold of control, t = 5.77, P < 0.01). In addition, calcitriol alone or in combination with PZA significantly enhanced the transcriptional level of anti-microbial peptides (cathelicidin LL-37: calcitriol vs. saline, 10.59 ± 1.03 vs. 2.80 ± 0.90 fold of control, t = 9.85, P < 0.01; mBD2: calcitriol vs. saline, 7.92 ± 0.62 vs. 1.79 ± 0.45 fold of control, t = 13.82, P < 0.01), whereas PZA exerted a negative effect on anti-microbial peptide gene expression.@*CONCLUSIONS@#Calcitriol as adjunctive treatment can result in beneficial treatment outcomes in M.tb infection by suppressing the inflammatory response and up-regulating the expression of anti-microbial peptides. These results indicate the feasibility of using calcitriol adjunctively with standard chemotherapy for the treatment of M.tb infection.
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Objective To analyze the differences of immune responses against Mycobacterium tu-berculosis antigens induced in two different nonhuman primates and to provide rationales for the selection of suitable animal models for vaccine efficacy evaluation.Methods Expression of functional surface markers including CD69 and HLA-DR, the activation markers on CD4+and CD8+T cells from in rhesus macaques and cynomolgus monkeys were measured by flow cytometry analysis.PBMCs were isolated from rhesus ma-caques and cynomolgus monkeys with Mycobacterium tuberculosis infection and stimulated with PPD and pep-tide pools ( ESAT-6/CFP-10) .Enzyme-linked imunospot ( ELISPOT) assay was performed to detect IFN-γproducing lymphocytes.Results The CD4+and CD8+T cells isolated from rhesus macaques without Myco-bacterium tuberculosis infection expressed higher levels of CD69 and HLA-DR than those from healthy cyno-molgus monkeys (P<0.01).The numbers of IFN-γspot forming cells/106 PBMCs in rhesus macaques with Mycobacterium tuberculosis infection for 10 and 11 months were respectively 3 and 3.5 times higher than that of cynomolgus monkeys upon after the stimulation of PBMCs with PPD.The levels of IFN-γproduction by the cells from rhesus macaque group were also higher than those from cynomolgus monkey group upon after the stimulation of PBMCs with ESAT-6 or CFP-10 peptide pools.Conclusion More IFN-γproducing cells were induced in rhesus macaques than that in cynimolgus monkeys after stimulation with Mycobacterium tu-berculosis antigens.Therefore, the rhesus macaques might be a better animal model for evaluating immune responses induced by Mycobacterium tuberculosis vaccines.
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The effect of the complement C1q expression on total hepatic ischemia-reperfusion (I/R) injury in rats was investigated. Sixty healthy male Sprague Dawley (SD) rats weighing 180-200 g were randomly divided into 5 groups: sham-operation group (S group, n=12); group of I/R for 1 h (I/R 1 h group, n=12); group of I/R for 3 h (I/R 3 h group, n=12); group of I/R for 6 h (I/R 6 h group, n=12); group of I/R for 24 h (I/R 24 h group, n=12). The hepatic I/R model of rats was established, and liver tissues were obtained 1 h, 3 h, 6 h and 24 h after hepatic I/R, respectively. Furthermore, the tissues were stained using hematoxylin-eosin, and the liver injuries of rats were observed using a microscope. The malondialdehyde (MDA) level and superoxide dismutase (SOD) activity in liver tissue were determined. Real-time polymerase chain reaction (PCR) and Western blotting were used to detect the expression levels of C1q mRNA and protein, respectively. As compared with the S group, the histopathological changes in I/R 1 h-24 h groups were gradually aggravated with the extension of I/R time. As compared with the S group, SOD activity and MDA content in the I/R groups were reduced and increased respectively with the extension of I/R time (P<0.01). Furthermore, the C1q expression at mRNA and protein levels in the I/R groups (especially in the I/R 3 h group) was significantly higher than that in the S group (P<0.05). It is suggested that C1q expression may play a principal role in hepatic I/R injury, particularly at the early stage of perfusion.
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The effect of the complement C1q expression on total hepatic ischemia-reperfusion (I/R) injury in rats was investigated. Sixty healthy male Sprague Dawley (SD) rats weighing 180-200 g were randomly divided into 5 groups: sham-operation group (S group, n=12); group of I/R for 1 h (I/R 1 h group, n=12); group of I/R for 3 h (I/R 3 h group, n=12); group of I/R for 6 h (I/R 6 h group, n=12); group of I/R for 24 h (I/R 24 h group, n=12). The hepatic I/R model of rats was established, and liver tissues were obtained 1 h, 3 h, 6 h and 24 h after hepatic I/R, respectively. Furthermore, the tissues were stained using hematoxylin-eosin, and the liver injuries of rats were observed using a microscope. The malondialdehyde (MDA) level and superoxide dismutase (SOD) activity in liver tissue were determined. Real-time polymerase chain reaction (PCR) and Western blotting were used to detect the expression levels of C1q mRNA and protein, respectively. As compared with the S group, the histopathological changes in I/R 1 h-24 h groups were gradually aggravated with the extension of I/R time. As compared with the S group, SOD activity and MDA content in the I/R groups were reduced and increased respectively with the extension of I/R time (P<0.01). Furthermore, the C1q expression at mRNA and protein levels in the I/R groups (especially in the I/R 3 h group) was significantly higher than that in the S group (P<0.05). It is suggested that C1q expression may play a principal role in hepatic I/R injury, particularly at the early stage of perfusion.
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Animals , Male , Rats , Blotting, Western , Complement C1q , Genetics , Metabolism , Gene Expression , Liver , Metabolism , Malondialdehyde , Metabolism , Random Allocation , Rats, Sprague-Dawley , Reperfusion Injury , Reverse Transcriptase Polymerase Chain Reaction , Superoxide Dismutase , Metabolism , Time FactorsABSTRACT
Objective To investigate the value of ultrasound-guided liver biopsy for infantile hepatitis syndrome regarding diagnosis,treatment and prognosis.Methods Fifty children with infantile hepatitis syndrome hospitalized in Guiyang Maternal and Child Hospital during Aug.2010 to May 2012 were involved in this study.Ultrasoundguided liver biopsies were performed to evaluate the inflammation grade and fibrosis stage.Immunohistochemical staining was used for pathogen diagnosis.The clinical outcomes were followed-up.Results Thirty-four cases (68％)were CMV infection,6 cases(12％) were vanishing bile duct syndrome,4 cases(8％) were chronic intrauterine infection,4 cases(8％) were congenital anomaly of bilirubin metabolism,and 2 cases (4％) were obstructive cholangitis.All 50 cases showed mild inflammation at portal area(G1-G2 grade).All 50 cases exhibited liver fibrosis.Sixteen cases were S1 stage,20 cases were S2 stage,8 cases were S3 stage and 6 cases were S4 stage.Pathogen analysis:all 50 cases showed intrahepatic cholestasis:38 cases were diffuse cholestasis,and 12 cases were moderate cholestasis.Treatment:all cases were treated using 2-week heteropathy; antivirus was used for CMV infected cases,thus 39 cases were finally cured,9 cases were relieved,and 2 cases were ineffective.Conclusion Liver biopsy is valuable for diagnosis,treatment and prognosis infantile hepatitis syndrome.
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Objective To evaluate the role of the complement 1 q (C1 q) in hepatic ischemia-reperfusion (I/R) injury in rats.Methods Sixty healthy male Sprague-Dawley rats,aged 3-4 months,weighing 180-200 g,were randomized into 2 groups:sham operation group (S group,n =12) and hepatic I/R group (I/R group,n =48).Hepatic specimens were obtained at 1,3,6 and 24 h of reperfusion and were then cut and stained with haematoxylin and eosin for examination of histological changes of the liver (with light microscope) and for determination of superoxide dismutase (SOD) activity and malondialdehyde (MDA) content (by colorimetric method),expression of Clq mRNA (using real-time PCR) and expression of Clq (by using Western blot).Results Compared with S group,the activity of SOD was gradually decreased,the content of MDA was gradually increased,and the expression of Clq and Clq mRNA was gradually up-regulated and peaked at 3 h of reperfusion with the prolongation of reperfusion time in I/R group (P ＜ 0.05).The pathological changes of the liver were aggravated with the prolongation of reperfusion time in I/R group.Conclusion Activation of C1 q is involved in hepatic I/R injury in rats.
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ObjectiveTo investigate the effect of flurbiprofen axetil on lung ischemia-reperfusion(I/R) injury in rats.MethodsSixty healthy male SD rats weighing 250-300 g were randomly divided into 3 groups( n =20 each): group sham operation(group S) ;group I/R and group flurbiprofen axetil (group FA).The animals were anesthetized with intraperitoneal 2％ pentobarbital 50 mg/kg and tracheostomized and mechanically ventilated.Lung I/R was induced by 60 min occlusion of left hilus pulmonis followed by 120 min reperfusion.In FA group flurbiprofen axetil 10 mg/kg was injected iv at 15 min before occlusion of left hilus pulmonis.The rats were sacrificed at 120 min of reperfusion and then the lungs were removed for measurement of lung wet/dry weight ratio,apoptosis index,NF-κB activity,Bcl-2 and Bax protien expression,and microscopic examination.Bcl-2/Bax ratio was caculated.ResultsI/R significantly increased lung wet/dry weight ratio,apoptosis index,NF-κB activity,Bcl-2 and Bax protien expression,and decreased Bcl-2/Bax ratio.Flurbiprofen axetil preconditioning significantly attenuated the I/R-induced changes in lung wet/dry weight ratio,apoptosis index,NF-κB activity,Bcl-2 and Bax protien expression,and Bcl-2/Bax ratio in group FA as compared with group I/R.Flurbiprofen axetil preconditioning also ameliorated I/R-induced lung damage.ConclusionFlurbiprofen axetil can attenuate lung I/R injury in rats by inhibiting NF-κB activity,up-regulating Bcl-2 expression and down-regulating Bax expression and inhibiting apoptosis.
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Objective To investigate the protective effects of heme oxygenase-1 (HO-1) mediated by cell penetrating peptide PEP-1 on myocardium against ischemia/reperfusion (IR) injury in isolated rat hearts. Methods Healthy male SD rats weighing 220-280 g were anesthetized with intraperitoneal pentobarbital. Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5% CO2 at 37 ℃. Eighteen isolated rat hearts were randomly divided into 3 groups ( n = 6 each): Ⅰ group sham operation (group S);Ⅱ group IR and Ⅲ group PEP-1/HO-1 + IR (group HO-1). The isolated rat hearts were perfused with an oxygena-ted (95% O2-5% CO2 ) K-H solution at 37 ℃ in a Langendorff apparatus and were subjected to 40 min of global ischemia followed by 50 min of reperfusion after 30 min of stabilization. In group Ⅲ (group HO- 1 ) the isolated hearts were perfused with 50 μmol/L PEP-1/HO-1 for 15 min before ischemia. After 50 min of reperfusion, HO-1expression, MDA content and SOD activity in myocardial tissues were determined. The activities of creatine kinase (CK) and lactic dehydrogenase (LDH) in coronary effluent fluid were measured. Results The HO- 1 expression was significanfly higher in HO-1 group than in group IR. IR induced significant increase in MDA content and decrease in SOD activity in myocardium and CK and LDH activities in coronary effluent in group Ⅱ compared with group S. PEP-1/HO-1 significantly attenuated IR-induced changes. Conciusion HO-1 mediated by PEP-1 has protective effects on myocardium ngainst IR injury in rats.
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Objective To investigate the effects of adenosine postconditioning (AP) on serum IL-10 and TNF-α concentrations following myocardial ischemia-reperfusion(VR)in rats.Methods Twenty-four SD ratsweighing 180-250 g were randomly divided into 4 groups(n=6 each):group I sham operation (group S);group Ⅱ myocardial I/R;group Ⅲ ischemic postconditioning(group IP)and group Ⅳ AP.Myocardial I/R was induced by 30 rain occlusion of anterior descending branch of left coronary artery followed by 120 min reperfnsion.IP was induced by 3 cycles of 30 s myocardial ischemia followed by 30 s reperfusion at the end of ischemia.In AP group adenosine 1.5 mg/kg was infused at 40μg·kg-1·min-1 before the onset of reperfusion.SP,DP and HR were recorded before ischemia (baseline) at 30 min of ischemia and 30 and 120 min of reperfusion.Arterial bloodsarnples were collected at 120 min of repednsion for determination of serum TNF-α and IL-10 concentrations.Theanimals were then killed.Their hearts were removed for microscopic examination.Myocardial infarct size wasmeasured and myocardial MDA content was determined.Results BP and HR were signilicandy decreased duringreperfusion while myocardial infarct size.MDA content and serum concentrations of IL-10 and TNF-α weresignificantly increased in I/R group compared with group S.Ischemic and adenosine postconditioning significantlyattenuated hypotension,reduced infarct size,myocardial MDA content and serum TNF-α concentration and increased serum IL-10 concentration in group AP and IP as compared with I/R group.There was no significant difference in the above changes between group AP and IP. Myocardial injury was ameliorated in group AP and IP as compared with I/R group. Conclusion Adenosine postconditioning can protect myocardium from I/R injury by increasing IL-10 production and inhibiting TNF-a release.
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Objective To investigate the acute toxicity of intravenous isoflurane in Beagles.Methods Six healthy adult Beagles of both sexes aged 6-8 months weighing 6-8 kg were used in this study.Isoflurane injectio (120 mg/ml) in 30% hpid emulsion was injected intravenously. Femoral artery was cannulated for direct BP monitoring.ECG was continuously monitored.The maximal tolerance dose (MTD) and approximate lethal dose (ALD) were determined by up-and-down technique. The initial dose was 3.0 ml/kg. The dose was decreased/increased by 0.3 ml/kg if the previous animal died/survived.The survived dogs were observed for 2 weeks.Autopsy and histopathological examination were performed on all dead Beagles.Results The ALD and MTD of intravenous isoflurane were 252 and 216 mg/kg. Autopsy and histopathological examination did not show any abnormality.Conclusion Cardiopulmonary depression is the main manifestation of the acute toxicity of intravenous isoflurane in Beagles.
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Objective To investigate the effect of PEP-1-heme oxygenase-1 (PEP-1-HO-1) fusion protein transduction on hypoxia-reoxygenation (H/R) injury in rat H9c2 cells. Methods After construction of the prokaryotic expression plasmid pET15b-PEP-1-hHO-1 containing the human heme oxygenase-1 gene, it was then transformed to make PEP-1-HO-1 fusion protein express. The H9c2 cells were cultured in high-glucose Dulbecco's modified Eagle's medium (DMEM) supplemented with 15% fetal bovine serum and randomly divided into 4 groups (n = 4 each): control group (group C), H/R group, low-concentration fusion protein group (group L-HO), and high-concentration fusion protein group (group H-HO). The cells were exposed to 22 h of hypoxia followed by 8 h of reoxygenation. PEP-1-HO-1 fusion protein was added to the culture medium with a final concentration of 1.0 μ mol/L (group L-HO) or 2.0 μmol/L (group H-HO) before hypoxia. The cells and supernatant of the culture medium were collected after reoxygenation to determine the activity of lactate dehydrogenase (LDH) in the supernatant and the content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) in the cells. Results The SOD activity was significantly lower, while the MDA content and LDH activity were significantly higher in group H/R, L-HO and H-HO than in group C (P ＜0.05). The SOD activity was significantly higher, while MDA content and LDH activity were significantly lower in group L-HO and H-HO than in group H/R, and in group H-HO than in group L-HO ( P ＜ 0.05). Conclusion PEP-1-HO-1 fusion protein transdution can protect H9c2 cells against H/R injury in rats.
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Objective To investigate the effects of emulsified isoflurane preconditioning on myocardial NF-κB activity during ischemia-reperfusion(I/R)in rats.Methods Forty-eight healthy male SD rats weighing 230-280 g were randomly divided into 4 groups with 12 animals in each group: sham operation group(group S),I/R group,lipid emulsion + I/R group(group L)and emulsified isoflurane + I/R group(group EI).In group I/R,EI and L,myocardial I/R was produced by occlusion of left coronary anterior descending artery(LAD)for 30 min followed by 120 min reperfusion.In group L,30% lipid emulsion 4 ml·kg-1 ·h-1 was infused intravenously for 30 min before myocardial I/R.In group EI,emulsified isoflurane 4 ml· kg- 1 · h- 1 was infused intravenously for 30 min followed by 15 min washout before myocardial I/R.In group S and I/R,normal saline was given instead.Blood samples were taken from femoral artery at the end of 120 min reperfusion for determination of serum cTnI and IL-6 concentrations and CK-MB activity by ELISA.The rats were then killed and the myocardial tissues were taken for determination of NF-κB activity by Western blot and observation of the ultrastructure by electron microscopy.Results The NF-κB activity,serum cTnI and IL-6 concentrations and CK-MB activity were significantly higher in group I/R,EI and L than in group S(P ＜ 0.05 or 0.01),while lower in group EI than in group I/R(P ＜ 0.05).Microscopic examination showed that emulsified isoflurane significantly attenuated the histopathological changes in group EI.Conclusion Emulsified isoflurane pretreatment can attenuate myocardial I/R injury through decreasing the NF-κB activity and inhibiting inflammatory response in rats.
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<p><b>OBJECTIVE</b>To study the BRCA1 mutations in patients with early-onset breast cancer and their affected relatives in Guangdong province and explore the relationship between BRCA1 mutation and the expressions of estrogen receptor(ER), progesterone receptor(PR), HER2 and ALN.</p><p><b>METHODS</b>From 58 patients with early-onset breast cancer and their affected relatives, the genomic DNA was extracted from the peripheral blood mononuclear cells and the coding regions of the BRCA1 gene was amplified using polymerase chain reaction. BRCA1 gene mutations were screened by denaturing high performance liquid chromatography (DHPLC) and subsequent direct DNA sequencing. The expression of ER, PR, HER2 and ALN were detected with immunohistochemistry and their relations with the gene mutation were analyzed.</p><p><b>RESULTS</b>Disease-related BRCA1 mutations were detected in 2 of the 58 patients, who were younger than 35 years old, including 1 with a novel splice-site mutation (IVS5-1 G-->A). No association was found between this novel mutation and the expressions of ER, PR, HER2 and ALN.</p><p><b>CONCLUSION</b>The incidence of BRCA1 mutation is significantly lower in patients with early-onset breast cancer and their affected relatives in Guangdong province than in the Western populations. The novel mutation identified in BRCA1 gene may represent a mutation characteristic of the patients in Guangdong province. BRCA1 gene mutations may not have any relation with the expression of ER, PR, HER2 and ALN.</p>
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Adult , Female , Humans , Age of Onset , Base Sequence , Breast Neoplasms , Genetics , China , DNA Mutational Analysis , Genes, BRCA1 , Genotype , Molecular Sequence Data , Mutation , Receptor, ErbB-2 , Genetics , Receptors, Estrogen , Genetics , Receptors, Progesterone , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To evaluate the protective effects of 8% emulsified isoflurane after myocardial ischemia-reperfusion injury and its mechanism in rabbits.</p><p><b>METHODS</b>Twenty-four male adult New Zealand white rabbits were anesthetized with intravenous injection of 30 mg/kg pentobarbital followed by 5 mg x kg(-1) x h(-1) infusion. All rabbits were subjected to 30 minutes of left anterior descending coronary artery (LAD) occlusion and 3 hours of subsequent reperfusion. Before LAD occlusion, the rabbits were randomly allocated into three groups for preconditioning treatment (eight for each group). The control group (C group) received intravenously 0.9% NaCl for 30 minutes. The emulsified isoflurane group (EI group) received 8% emulsified isoflurane intravenously till 0.64% end-tidal concentration for 30 minutes that was followed by a 15-minute washout period. The Intralipid group (IN group) received 30% Intralipid for 30 minutes. The infarcted area, plasma malondialdehyde (MDA) content, superoxide dismutase activity (SOD) and nitrite concentration after 3-hour myocardial perfusion were recorded simultaneously.</p><p><b>RESULTS</b>For the myocardial ischemia-reperfusion injury animals, the infarcted size in the EI group was significantly reduced (91.9% +/- 8%) as compared with control group (39% +/- 6%, t=5.19, P<0.01). The plasma SOD activity and nitrite concentration in EI group were significantly higher than those in control group (t=2.82, t=8.46, P<0.05), but MDA content was lower in EI group than that in control group (t=2.56, P<0.05).</p><p><b>CONCLUSIONS</b>The results indicate that emulsified isoflurane has a cardioprotection effect against ischemia-reperfusion injury. This beneficial effect of emulsified isoflurane is probably through NO release and consequently by increase in antioxidation of myocardium.</p>
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Animals , Male , Rabbits , Emulsions , Isoflurane , Pharmacology , Lipid Peroxidation , Myocardial Infarction , Drug Therapy , Pathology , Myocardial Reperfusion Injury , Nitric Oxide , Blood , Superoxide Dismutase , MetabolismABSTRACT
Objective To explore the way and technique to gain adequate seed cells for skin tissue engineering sufficiently. Methods Human telomerase reverse transcriptase (hTERT) gene was introduced into rabbit keratinocytes by eukaryotic vector. The positive clones were selected and cultured in microcarrier-RCCS. The growth of immortalized keratinocytes was observed, and the metabolic rate and pan-cytokeratins (AE1/AE3) expression of immortalized keratinocytes in experimental groups were detected and compared with those in the control group. Results The immortalized keratinocytes in the experimental groups grew rapidly and had a high metabolic rate and shorter population doubling time (PD) (P
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<p><b>AIM</b>To study the changes of synaptic plasticity in rat model with Alzheimer disease (AD).</p><p><b>METHODS</b>AD rat model was conducted by D-galactose intraperitoneal injection combined with lesions of nucleus basalis of Meynert (nbM). Behavioral performance, LTP in dentate gyrus and synaptic morphology in hippocampal CA1 were observed.</p><p><b>RESULTS</b>(1) Escape latencies in place test in model rats were longer than that in control rats, and swimming time and distance between the two groups in platform quadrant were significant differently (P < 0.01). (2) The numerical density (Nu) and surface density (Su) of synaptic contact zones markedly decreased (P < 0.01) in model rats. (3) Augment of population spike (PS) in perforant path-dentate gyrus of model rats after high frequency stimulation was smaller than that of the control (P < 0.05).</p><p><b>CONCLUSION</b>The results suggest that the decreased synaptic plasticity in hippocampus could responsible for the impairment of spatial learning of model rats.</p>
Subject(s)
Animals , Female , Male , Rats , Alzheimer Disease , Basal Nucleus of Meynert , Pathology , Disease Models, Animal , Galactose , Pharmacology , Hippocampus , Long-Term Potentiation , Neuronal Plasticity , Rats, WistarABSTRACT
Objective To observe the effect of Bushen Yizhi Prescription(BYP) on long-term potentiation (LTP) in hippocampus of ovariectomized (OVX) rats. Methods Three-month-old female SD rats were randomized into mimic operation group, model group, low dosage BYP group, high dosage BYP group and estrogen control group. Ovariectomy was operated in the three latter groups. After operation, BYP group was treated with gastric infusion of BYP and estrogen group with subcutaneous injection of estrogen. The changes of LTP of dentate gyms in hippocampus of OVX rats were observed by electrophysiological method and the effect of BYP on LTP was also evaluated. Result & Conclusion The increased amplitude of population spike of LTP was significantly low and maintained a short time in model group as compared with that in mimic operation group. There were insignificant difference between the high-dosage BYP group and estrogen control group. It is indicated that synaptic transmission in hippocampus can be protected and sustained by BYP and the estrogen replacement.
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Objective: In order to further investigate the structural/functional relationship of TRALL. Methods: We did homology modeling for the extracellular segment of TRAIL, which is from R117 to G281, totally 165 aa residues long. The modeling software is Insight II from MSI/Biosym and the modeling work is based on the three dimensional structure of TNF-?. Results: From the modeling result, it can be seen that the modeled structure of TRAIL contains 10 ?-sheets and homologs for all these sheets could be found in TNF-?. This just confirms with the principle that the structurally con-seived regions within molecules of the same structure family should experience relatively small sequence mutations. In addition, the credibility of the modeled structure is checked by the way of inverse folding from Profile-3D. Conclusion: The result shows modeled structure is generally correct.
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Objective To determine blood/gas and tissue/gas partition coefficients of sevoflurane, isoflurane and halothane and evaluate the effects of pregnancy on them. Methods Ten 18-22 day pregnant and 10 non-pregnant SD rats were killed under pentobarbital anesthesia. The tissue specimens of heart, liver, kidney and brain were obtained and made into homogenates respectively. The blood/gas and tissue/gas partition coefficients of sevoflurane, isoflurane and halothane were determined using a method of 2-stage headspace equilibration by gas chromatography. Results Blood/gas and brain/gas partition coefficients were lower in pregnant group than in non-pregnant group (P