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@#【Objective】To evaluate the safety and effects of transcatheter arterial chemoembolization using Drug-eluted beads（DEB-TACE）plus apatinib in patients with hepatocellular carcinoma（HCC）.【Methods】A retrospective analysis was performed，which included 11 HCC patients treated with DEB- TACE and followed by a target therapy of apatinib（500 mg QD）in our clinical research center. Radiograph evaluation and tumor biomarker，alpha- fetoprotein（AFP）， were recorded before the procedure and during the follow-up of the first cycle after 4~8 weeks. Adverse events induced by apatinib were recorded. 【Results】 According to the response evaluation criteria in solid tumors （RECIST），modified response evaluation criteria in solid tumors （mRECIST） and European Association for the study of the liver （EASL） criteria，the objective response rate （ORR） was 36.4% ，63.6% ，72.7% respectively，and the disease control rate（DCR）was 90.7% ，72.7% ，81.8% respectively. AFP levels were 44 251.7 μg/L which significantly decreased after the procedure ，compared to 366 336 μ g/L ，the levels before the treatment. Four reversible grade III adverse events were recorded and no grade IV adverse events found in these cases. 【Conclusion】According to the short-term treatment response and safety，DEB- TACE combined with apatinib could be considered as a promising treatment for intermediate and advanced stage hepatoma.
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@#【Objective】To explore the correlation between the expression levels of copper chaperone for superoxide dismutase（CCS）and the malignancy related biomarkers in hepatocellular carcinoma（HCC）.【Methods】From January to December 2018，we obtained fresh samples of surgically dissected HCC paired with para-carcinoma normal tissues from 10 HCC patients and collected their clinical and pathological data. Western blotting（WB）was performed to examine the expression of CCS in HCC tissues and adjacent noncancerous tissues. Immunohistochemical staining（IHC）was employed to detect the expression of Ki67，CD34，vimentin and glypican-3（GPC3）. The correlation between the expression levels of CCS and biomarkers was analyzed by using Wilcoxon rank sum test. The association between CCS expression and clinical pathological characteristics of HCC patients was investigated by using Fisher′s exact probability test.【Results】In 7 of the 10 HCC cases，the expression level of CCS in HCC tissue samples was lower than that in adjacent noncancerous tissues，and in other 3 HCC cases，CCS expression higher. In the group with low CCS expression，compared with those in the group with high CCS expression，the expression levels of Ki67，vimentin and GPC3 were higher （Z=- 2.400，P=0.016；Z=- 2.423，P=0.015；Z=- 2.400，P=0.016），while the expression level of CD34 lower（Z=- 2.423，P=0.015）. There was no statistically significant difference in clinical and pathological variables including gender ，age ，hepatitis B virus infection，liver cirrhosis，preoperative serum AFP level，tumor size，Edmondson-Steiner grade and microvascular invasion between two groups with high and low CCS expression.【Conclusions】The results revealed that in most of HCC patients，the expression level of CCS in HCC tissues was lower than that in adjacent noncancerous tissues. Additionally ， higher expression levels of Ki67，vimentin and GPC3 in HCC tissues with low CCS expression indicated that low expression level of CCS correlated with malignant biological behaviors such as HCC proliferation ，invasion and metastasis. The mechanism that the expression level of CD34 appeared lower in HCC tissues with low CCS expression，however，needs further study. These findings suggest that compared with that in normal liver tissues，CCS expression is decreased in a majority of the cases，and it may serve as a promising therapeutic and prognostic biomarker for HCC.
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<p><b>OBJECTIVE</b>This study is to examine the influence of familiarity on energy intake, eating behavior, and concentration of the plasma gut hormones in lean and overweight young male subjects.</p><p><b>METHODS</b>Twenty-eight lean and twenty-eight overweight participants were recruited. Their food consumption was documented and analyzed when they had a test meal while they were paired with friends or strangers at the same weight stature. Their eating behavior was recorded with cameras hidden in the carton, and postprandial plasma gut hormone concentration were measured.</p><p><b>RESULTS</b>Compared with overweight strangers (OS), overweight friends (OF) had increased food consumption, prolonged and decreased number of chews per 10 g food. Compared with OS, postprandial plasma concentration of cholecystokinin-8 was significantly lower in OF group at 30, 60, and 90 min, whereas the concentration of glucagon-like peptide 1 was significantly lower at 60 and 90 min. Plasma ghrelin concentration was significantly higher in the OF group than that in the OS group at 90 and 120 min. No significant differences in gut hormone concentration were observed between lean strangers (LS) and lean friends (LF) groups at all time points.</p><p><b>CONCLUSION</b>Familiarity plays an important role in increasing energy intake and in changing of postprandial gut hormone concentration in overweight individuals.</p>
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<p><b>OBJECTIVE</b>To investigate the effect of P2X7R antagonist brilliant blue G (BBG) on aGVDH of mice after allo-HSCT.</p><p><b>METHODS</b>aGVHD mouse model after HSCT was established and treated with the P2X7R antagonist BBG of different dosages (50 mg/kg and 75 mg/kg). After treatment, the survival, body weight, pathological and liver function of aGVDH mice were abserved, and the expression levels of P2X7, NLRP3, caspase-1, IL-1β, IL-18 mRNA and protein were evaluated by real-time PCR and Western blot.</p><p><b>RESULTS</b>The allo-HSCT aGVHD mouse model was successfully established, the intraperitoneal injection of BBG alleviated the aGVHD clinical manifestations including roachback, ruffled fur, skin peeling and weight loss of recipient mice, decreased P2X7R and IL-1β expression and reduced the mRNA levels of P2X7R, NLRP3, Caspase-1, IL-1β and IL-18. Furthermore, GVHD group receiving 75 mg/kg BBG showed most significant difference of these indexes.</p><p><b>CONCLUSION</b>BBG alleviates liver inflammatory damage induced by aGVHD after allo-HSCT, and decreases the expression of proinflammatory cytokines. Moreover, the protective effect of that of BBG 75 mg/kg group is better than that of BBG 50 mg/kg group.</p>
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Objective To evaluate clinical significance of direct antiglobulin testing(DAT)in anemia in patients with severe chronic hepatitis B(CHB).Methods Red blood cell(RBC)-related parameters detection and DAT were performed among 30 healthy persons,30 CHB patients,and 50 severe CHB patients,clinical factors related to posi-tive DAT were analyzed.Results RBC count,hemoglobin (Hb)concentration,and hematocrit(HCT)level in severe CHB patients were all lower than CHB patients and healthy group(P <0.05),RBC distribution width(RDW)in severe CHB patients were all higher than CHB patients and healthy group(P<0.05);the positive rate of DAT in patients with se-vere CHB,CHB,and healthy group were 62.82%,13.33% and 0 respectively.RBC count,Hb concentration,and HCT level in severe CHB patients with positive DAT were all lower than severe CHB patients with negative DAT (all P <0.05),while RDW was higher than the latter (P=0.001);after RBC was separated through capillary,positive intensity of DAT of aged RBCs was higher than young RBCs in severe CHB patients (P <0.001);among severe CHB patients, DAT-positive and-negative patients differed in gender,age,alanine aminotransferase,total bilirubin,complement C3, C-reactive protein,and complication of diabetes(all P≤0.05).Conclusion Anemia in severe CHB patients may be re-lated to immune hemolysis of aged RBCs induced by antibody adsorption.
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Objective:To preliminarily verify the cross talk between tissue factor/active coagulation factor Ⅶ (TF/FⅦa) and epidermal growth factor receptor (EGFR) pathways in human colon cancer cells in culture.Methods:FⅦa was treated to HT-29 (KRAS-wild type) and LoVo (KRAS-mutant) colon cancer cells to activate TF/F Ⅶa pathway,qRT-PCR and Western blot were used to detect the expressions of amphiregulin (AREG) and epiregulin (EREG),ligands of EGFR on mRNA and protein levels,respectively.After knocking down expression of TF by TF-targeted siRNA transfection,FⅦa was treated and mRNA expressions of AREG and EREG were detected to see whether the FⅦa-induced effects were dependent on TF.Expressions of mRNA of TF and FⅦwere detected by qRT-PCR following the activation of EGFR pathway by treatment with epidermal growth factor (EGF) to HT-29 and LoVo cells.Results:After TF/FⅦa pathway was activated,for HT-29 cells,expressions of AREG (on mRNA level) and EREG (both on mRNA and protein level) were significantly down-regulated versus those of control group,gene expressions of AREG and EREG were 0.55 ± 0.09 vs.0.99 ± 0.09,0.67 ± 0.10 vs.1.02 ± 0.02,protein expressions of EREG were 0.54 ± 0.09 vs.1.04 ± 0.13,all P ＜ 0.05.For LoVo cells,expressions of AREG (both on mRNA and protein level) and EREG (on protein level) were significantly up-regulated versus those of control group,gene expression of AREG were 1.87 ± 0.39 vs.0.93 ± 0.23,protein expressions of AREG and EREG were 3.09 ±0.73 vs.1.11 ±0.21,1.53 ±0.19 vs.0.97 ± 0.23,all P ＜0.05.The regulating effect of AREG and EREG mRNA expression by FⅦa in HT-29 and LoVo cells could both be partly blocked by knocking down TF expression.For HT-29 cells,activation of EGFR pathway induced no significant TF mRNA expression,F Ⅶ mRNA expression was not detected.However,for LoVo cells,activation of EGFR pathway induced significantly higher mRNA expressions of both TF and FⅦ,expressions were 1.53 ± 0.23 vs.1.00 ± 0.23,53.20 ± 6.08 vs.1.00 ± 0.15,all P ＜0.05.Conclusion:In colon cancer cell LoVo,when activated,TF/FⅦa pathway and EGFR pathway could interact through upregulating the other pathway's effectors,and mutant KRAS might play a critical role in the two pathways'cross talk.
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Objective To evaluate clinical significance of direct antiglobulin testing(DAT)in anemia in patients with severe chronic hepatitis B(CHB).Methods Red blood cell(RBC)-related parameters detection and DAT were performed among 30 healthy persons,30 CHB patients,and 50 severe CHB patients,clinical factors related to posi-tive DAT were analyzed.Results RBC count,hemoglobin (Hb)concentration,and hematocrit(HCT)level in severe CHB patients were all lower than CHB patients and healthy group(P <0.05),RBC distribution width(RDW)in severe CHB patients were all higher than CHB patients and healthy group(P<0.05);the positive rate of DAT in patients with se-vere CHB,CHB,and healthy group were 62.82%,13.33% and 0 respectively.RBC count,Hb concentration,and HCT level in severe CHB patients with positive DAT were all lower than severe CHB patients with negative DAT (all P <0.05),while RDW was higher than the latter (P=0.001);after RBC was separated through capillary,positive intensity of DAT of aged RBCs was higher than young RBCs in severe CHB patients (P <0.001);among severe CHB patients, DAT-positive and-negative patients differed in gender,age,alanine aminotransferase,total bilirubin,complement C3, C-reactive protein,and complication of diabetes(all P≤0.05).Conclusion Anemia in severe CHB patients may be re-lated to immune hemolysis of aged RBCs induced by antibody adsorption.
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<p><b>OBJECTIVE</b>To investigate the protective effects of polydatin on sepsis-induced acute liver injury (ALI) in mice, and to preliminarily study its mechanisms.</p><p><b>METHODS</b>The sepsis model was established using cecal ligation and puncture (CLP).A sham-operation control group was also set up. Polydatin (50, 100, and 300 mg/kg, respectively) was administrated to mice 1 h before CLP. The survival and liver injury were evaluated subsequently per 6 h after CLP. The survived mice were scarified 24 h later. The serum and the liver tissue sample were collected. The serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected by colorimetric method. The content of tumor necrosis factor-alpha (TNF-alpha) was assayed by ELISA. The cyclooxygenase-2 (COX-2) expression in the liver tissue was detected by Western blot. The pathological changes of the hepatic tissue were analyzed by hematoxylin and eosin stain.</p><p><b>RESULTS</b>The mortality of mice reached as high as 50% at 24 h after CLP. The biochemical indices and the pathological changes of the liver tissue showed obvious lesion. The success rate of modeling was 90%. Compared with the sham-operation control group, the serum ALT,AST activity, the TNF-alpha content, and the hepatic COX-2 protein expression markedly increased in the CLP group (P < 0.01). Polydatin improved the sepsis-induced mortality dose-dependently, inhibited increased ALT, AST activity and TNF-alpha, decreased the hepatic COX-2 protein expression, and attenuated the pathological injury of the liver (P < 0.05).</p><p><b>CONCLUSION</b>Polydatin could effectively protect sepsis-induced ALI, which might be achieved possibly through inhibiting serum TNF-alpha production and hepatic COX-2 expression.</p>
Subject(s)
Animals , Mice , Alanine Transaminase , Blood , Aspartate Aminotransferases , Blood , Cyclooxygenase 2 , Metabolism , Disease Models, Animal , Glucosides , Pharmacology , Liver , Mice, Inbred Strains , Sepsis , Metabolism , Pathology , Stilbenes , Pharmacology , Tumor Necrosis Factor-alpha , BloodABSTRACT
This study was aimed to explore the contribution of autophagy associated gene Beclin1 in the prognosis of myelodysplastic syndrome (MDS) by detecting the expression level of Beclin1 in bone marrow mononuclear cells (BMNC) from 40 MDS patients, 14 non-malignant anemia patients and 25 AML patients. The expression of Beclin1 mRNA was detected by real-time quantitative polymerase chain reaction (qRT-PCR). At the same time, the Western blot was used to analyze the expression of Beclin1 proteins. The results showed that the expression of Beclin1 in low risk MDS patients and non-malignant anemia patients was both significantly higher than that in acute myeloid leukemia patients (P < 0.01). And more interestingly, the Beclin1 mRNA expression in MDS group was negatively correlated with World Health Organization classification-based prognostic system (WPSS) score (r = -0.495). It is concluded that the expression of Beclin1 in the patients with MDS is higher than that in AML patients, and negatively correlated with WPSS scores. Beclin1 is a potential biomarker for predicting prognosis of the patients with MDS.
Subject(s)
Humans , Anemia , Metabolism , Apoptosis Regulatory Proteins , Genetics , Metabolism , Autophagy , Beclin-1 , Biomarkers, Tumor , Metabolism , Bone Marrow Cells , Metabolism , Leukemia, Myeloid, Acute , Metabolism , Membrane Proteins , Genetics , Metabolism , Myelodysplastic Syndromes , Metabolism , Pathology , PrognosisABSTRACT
Objective To observe the protein and mRNA expression of thyroid-stimulating hormone receptor (TSHR) in mammary gland tissue of lactating rats,and to explore iodine uptake mechanism.Methods Eighty adult Wistar rats (60 female and 20 male),weighting 210-250 g were selected.All female Wistar rats were randomly divided into 6 groups according to their body mass:normal non-pregnant group,lactating for 5-,10-,15-and 20-day groups and weaning for 5 days group,10 rats in each group.All rats were fed with conventional fodder and tap water freely.In addition to the normal non-pregnant group,other five groups of female and male rats were mated at 3 ∶ 1,respectively.Then the rats in all groups were killed on the 5th,10th,15th and 20th day after lactation and on the 5th day after weaning to get the mammary gland tissue.The protein and mRNA expression of TSHR were determined by immunohistochemical staining and real-time quantitative PCR.Results TSHR protein was expressed in mammary acinar and ductal epithelial cytoplasm.The expression of TSHR in mammary gland showed significant differences between groups (x2 =14.612,P ＜ 0.05),the staining intensity of mammary gland tissue in normal non-pregnant rats(weak,n =4; moderate,n =6) was weaker than that of lactating for 5 days(weak,n =2; moderate,n =3; strong,n =5) and 10 days groups(barely detectable,n =1;moderate,n =4; strong,n =5; x2 =4.113,5.250,all P＜ 0.05).The expression of TSHR mRNA in mammary gland showed significant differences between groups(F=20.488,P ＜ 0.05); the expression of TSHR mRNA in lactating for 10 days group(0.31 ± 0.06) was higher than that of lactating for 5 days group(0.22 ± 0.04,P ＜ 0.01),and the expression of lactating for 15 days group (0.16 ± 0.08) was significantly lower than that of lactating for 5 days group (P ＜ 0.05).Conclusions TSHR is widely expressed in mammary gland of lactating rats.The iodine uptake of mammary gland is enhanced in early lactation period when the body may be more susceptible to iodine deficiency,therefore iodine should be supplemented reasonably.
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Objective To observe the effect of different levels of iodine nutrition on rat maternal thyroid function during pregnancy.Methods A total of 225 Wistar rats one month after weaning were involved in the study(female 165,male 60,body mass 80 to 100 g).Female rats were randomly divided into six groups by body mass:control group(NI group),iodine deficiency 1 and 2 groups(LI1,LI2 groups),iodine excess 1 and 2 groups (HI1,HI2 groups),and the control of not pregnant group(NNI group).There were 30 rats in 1-5 groups and 15 rats in group 6.LI1,LI2 groups:low iodine diet + deionized water of no iodine or iodine-containing 5 μg/L; HI1,HI2 groups:normal diet + deionized water of iodine 3000,10 000 μg/L; NI,NNI groups:normal diet + deionized water of iodine-containing 50 μg/L.After 12 weeks,the females(except group 6) mated the male by 2 ∶ 1,and then each pregnant female rat was fed in a single cage.The female mice were sacrificed in the first(5 ± 2)d,the second (12 ± 2)d and the third trimesters of pregnancy (17 ± 2)d,respectively,and there blood samples and thyroid were obtained.Serum total thyroxine(TT4),free thyroxine(FT4),total triiodothyronine (TT3),free triiodothyronine (FT3) and thyroid stimulating hormone(TSH) were determined by radioimmunoassay and serum thyroglobulin(TG) and thyroid-binding globulin (TBG) were determined by enzyme-linked immunosorbent assay.Results ①Thyroid absolute quality and relative quality was compared among groups,and the differences were statistically significant (F =16.55,24.25,F ＜ 0.01 or ＜ 0.05).②At the first,the second and the third trimesters of pregnancy,the differences of maternal serum TT4 and FT4 between groups were statistically significant(F =5.02,13.41,17.39,41.89,23.72,48.64,P ＜ 0.01 or ＜ 0.05).Female rats in NI,HI1 and HI2 groups in different pregnant periods among inner groups were compared,and the differences of serum TT4 and FT4 were statistically significant(F=3.27,6.98,8.22,8.65,29.68,7.90,P ＜ 0.01 or ＜ 0.05).③ In the first and the third trimesters of pregnancy,maternal serum TT3 was compared among groups,and the differences were statistically significant(F=3.59,8.22,P ＜ 0.05 or ＜ 0.01) ; in the second and the third trimesters of pregnancy,maternal serum FT3 was compared among groups,and the difference was statistically significant(F =3.86,4.26,P ＜ 0.05 or ＜ 0.01).Female rats in NI,LI1 and HI1 groups in different pregnant periods among inner groups were compared,and the differences of maternal serum TT3 were statistically significant(F =8.77,7.11,6.28,P ＜ 0.01 or ＜ 0.05).④At the first,the second and the third trimesters of pregnancy,the differences of maternal serum TG and TBG were compared in groups,and the differences were statistically significant(F =5.47,3.62,9.35,4.15,13.16,22.78,P ＜ 0.01 or ＜ 0.05).The differences of maternal serum TG of HI1 group and of serum TBG of NI group in different pregnant periods among inner groups were statistically significant (F =3.18,7.94,P ＜ 0.05).⑤At the first,the second and the third trimesters of pregnancy,the differences of maternal serum TSH in groups were statistically significant(F =4.83,7.08,6.52,P ＜ 0.01); the differences of maternal serum TSH of all the 5 groups in different pregnant periods among inner groups were statistically significant (F =3.26,8.89,11.45,4.04,3.78,P ＜ 0.05).Conclusions Different levels of iodine nutrition can cause changes in thyroid function in rats maternal thyroid function during pregnancy; serum TT4,FT4 level decreases when iodine deficiency,and increase with iodine excess.Serum TT3,FT3 level of does not changed significantly due to compensatory regulation of the body.
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Objective To study the effects of different iodine intakes on rat iodine metabolism during pregnancy.Methods One hundred and fifty female Wistar rats (body weight 80-100 g) were randomly divided into five groups:control group(NI),lower iodine 1 and 2 groups(LI1 and LI2),High iodine 1 and 2 groups(HI1 and HI2) by weight,30 rats in each group.These rats were given deionized water containing different concentrations of iodine,50(NI),0 (LI1),5(LI2),3000(HI1) and 10000 μg/L(HI2),respectively.After 12 weeks,urine samples were collected before copulation.The rats were sacrificed at the first(6-7 days),second (12-13 days) and third trimesters(19-20 days),respectively,serum and amniotic fluid samples were collected.Urinary iodine and iodine level in the fetal amniotic fluid were measured by As3+-Ce4+ catalytic spectrophotometry.Serum iodine was measured by mild acid digestion method.Results The baseline medians of urinary iodine of LI1 and LI2 groups(5.96,15.92 μg/L) were significantly lower than that of the NI group(43.75 μg/L,all P ＜ 0.01),and the values of HI and HI2 groups(5263.96,20389.64 μg/L) were significantly higher than that of the NI group (all P ＜ 0.01).The median of urinary iodine during pregnancy was significantly lower than that of the baseline of no pregnancy(all P ＜ 0.01).The medians of urinary iodine of the NI group at the first and the second trimesters (28.97,34.34 μg/L) were significantly lower than that of the third trimester(42.31 μg/L,all P ＜ 0.01).The means of serum iodine of LI1 and LI2 groups[(3.68 ± 1.69),(10.45 ± 4.16) μg/L] were significantly lower than that of the NI group [(23.68 ± 3.85)μg/L,all P ＜ 0.05],and the means of serum iodine of HI1 and HT2 groups [(502.67 ± 97.03),(822.15 ± 139.45)μg/L] were significantly higher than that of the NI group (all P ＜ 0.01).Although the mean of serum iodine of HI group gradually decreased with the progression of gestation,the difference was not statistically significant(all P ＞ 0.05).The iodine levels in amniotic fluid of fetal rats at the second and the third trimesters in LI1 group(0.85,3.00 μg/L) were significantly lower than that of the NI group(3.56,7.91 μg/L,all P ＜ 0.01),but the difference was not statistically significant between the iodine level in amniotic fluid of fetal rats of the LI2 and the NI groups at the second and the third trimesters(all P ＞ 0.05).The iodine levels in amniotic fluid of fetal rats at the second and the third trimesters in the HI1 group(49.59,171.21 μg/L) were significantly higher than that of the NI group(all P ＜ 0.01).The iodine levels in amniotic fluid of fetal rats at the second and the third trimesters in HI2 group (98.76,544.77 μg/L) were significantly higher than that of the NI group(all P ＜ 0.01).The iodine level in amniotic fluid of fetal rats in the third trimester was significantly higher than that of the second trimester in all the groups (all P ＜ 0.01).The ratios of serum iodine and urinary iodine of the LI1 and the LI2 groups (1.29 ± 1.14,1.70 ± 1.01) were significantly higher than that of the NI group(0.51 ± 0.37,all P ＜0.01),and that of the HI1 and the HI2 groups(0.21 ± 0.07,0.11 ± 0.07) were significantly lower than that of the NI group (all P ＜ 0.01).The ratios of amniotic fluid iodine and serum iodine of the LI and the LI2 groups (0.19 ± 0.15,0.32 ± 0.17) were significantly higher than that of the NI group(0.13 ± 0.05,P ＜ 0.01),but the difference was not statistically significant between HI1 and HI2 groups(0.09 ± 0.03,0.11 ± 0.04) and NI group(all P ＞ 0.05).The ratio of amniotic fluid iodine and serum iodine of the third trimester was significantly higher than that of the second trimester(all P ＜ 0.05).Conclusions Different iodine intake leads to changes in the levels of maternal iodine metabolism in rats during pregnancy.There probably is a protection mechanism in the mother's body,which protects the mother and the fetal from injury by iodine excess or iodine deficiency.
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ObjectiveTo study the mRNA expression of rat Insulin-like growth factors- Ⅰ (IGF- Ⅰ ) and Transforming growth factor-β1 (TGF-β1) in thyroid and placenta with different iodine intakes during pregnancy.MethodsOne hundred and fifty female Wistar rats,weighting 80 - 100 g,were randomly divided into five groups according to body weight,30 rats in each group.Each group was given deionized water containing different concentrations of iodine,50 μg/L(control group,NI),0 μg/L(iodine deficiency 1 group,LI1 ),5 μg/L(iodine deficiency 2 group,LI2),3000 μg/L(iodine excess 1 group,HI1 ),and 10 000 μg/L(iodine excess 2 group,HI2),respectively.After feeding for 12 weeks,the female rats were mated with male rats.The female rats were sacrificed at first(6,7 days),trimester( 12,13 days),and third trimesters( 19,20 days),respectively,then their thyroid and placenta were collected.The mRNA expressions of IGF- Ⅰ and TGF-1 in thyroid and placenta were detected by real-time quantitative PCR.Results①The actual thyroid weights of LI1 and LI2 groups[ (12.17 ± 5.41 ) × 10-2 g,(3.54 ± 1.21) × 10-2 g] were significantly higher than that of NI group[ (2.05 ± 0.50) × 10-2 g,all P ＜ 0.05] ;actual weights of HI1 and HI 2 groups[ (1.64 ± 0.27) × 10-2 g,(1.66 ± 0.29) × 10-2 g] were compared with that of NI group,the difference was not statistically significant(all P ＞ 0.05).②The mRNA expression of IGF- Ⅰ: at the first trimester,LI1 and LI2 groups(l.98 ± 0.35,1.47 ± 0.22) were all higher than that of NI group(1.01 ± 0.18,all P＜ 0.01 ),HI1 and HI2 groups(0.68 ± 0.16,0.75 ± 0.09) were lower than that of NI group(all P ＜ 0.01 );at the second trimester,HI2 group( 1.14 ± 0.17) was lower than that of NI group( 1.58 ± 0.33,P ＜ 0.01 ) ; at the third trimester,LI2 and HI2 groups(1.47 ± 0.20,1.45 ± 0.35) were lower than that of NI group(2.20 ± 0.37,all P＜0.01).The mRNA expression of IGF- I level in NI group at the first,second,and third trimesters(1.01 ±0.18,1.58 ±0.33,2.20 ± 0.37) was up regulated gradually,pairwise comparisons were statistically significant(all P ＜ 0,01 ).③The mRNA expression of TGF-β1: at the first trimester,LI1 group (1.37 ± 0.13) was higher than NI group (1.05 ±0.18,P ＜ 0.01 ),HI1 and HI2 groups(0.50 ± 0.09,0.44 ± 0.11) were lower than NI group(all P＜ 0.01); at the second trimester,LI1 and HI2 groups(1.39 ± 0.28,1.17 ± 0.12) were higher than NI group(0.63 ± 0.22,all P ＜0.01 ) ; at the third trimester,LI1 and LI2 groups ( 1.57 ± 0.30,1.23 ± 0.20) were higher than NI group ( 0.68 ± 0.17,all P＜ 0.01).TGF-β1 mRNA expressions of NI group at the second (0.63 ± 0.22) and third trimesters(0.68 ± 0.17) were lower than that of the first trimester (1.05 ± 0.18,all P ＜ 0.01).④ Rats' IGF-Ⅰ mRNA expression in placental: at the second trimester HI1 group,HI2 group( 1.48 ± 0.16,1.45 ± 0.25) were all higher than the NI group ( 1.00 ± 0.10,all P ＜ 0.01 ) ; at third trimester,HI1 group ( 1.75 ± 0.15 ) were higher than the NI group ( 1.54 ± 0.29,P＜ 0.05),HI2 group(l.94 ± 0.31) were higher than the NI group(P ＜ 0.01 ).IGF- Ⅰ mRNA expression in placental of NI group at the third trimester was higher than the second trimester(P＜ 0.01).⑤ Rats' TGF-β1 mRNA expression in the placenta: at the second trimester and the third trimester of pregnancy there were no significant difference between the five groups(all P ＞ 0.05) ; NI group at the third trimester(0.83 ± 0.16) was lower than the second trimester(0.98 ± 0.20,P ＜ 0.05).Conclusions During pregnancy,IGF- I mRNA expression increases in thyroid under the conditions of iodine deficiency,and this effect is particularly significant in the first trimester; at the same time,TGF-β1 mRNA expression is increased,and this inhibition becomes clear with the deepening of iodine deficiency.Under the condition of iodine excess,the functions of IGF- Ⅰ and TGF-β1 in thyroid above-mentioned were relatively weak.With the development of gestational period,promoting tissues growth and differentiation effect of placenta's IGF- Ⅰ was more significant gradually,but,inhibited effect of TGF-β1 was weaken.
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<p><b>OBJECTIVE</b>To investigate whether Resolvin E1 (RvE1) could protect against ox-LDL-induced injury on human vein vascular endothelial cells and reveal related molecular mechanisms.</p><p><b>METHODS</b>Human vein vascular endothelial cells were randomly assigned to six groups, which were treated with saline, RvE1, wortmanin, ox-LDL, ox-LDL and RvE1, ox-LDL and RvE1 and wortmanin, respectively. After 48 h, survival rates were determined by MTT, apoptosis rate of cells were determined by flow cytometry, TNF-α contents were assayed by ELISA, caspase 3 and 9 activities were measured by microplate reader, and the expression of p-AKT and LOX-1 were determined by Western blot.</p><p><b>RESULT</b>Compared with normal saline group, survival rate was markedly decreased and apoptosis rate, TNF-α content, caspase 3 and 9 activities, and the expression of LOX-1 were significantly increased in ox-LDL group (P < 0.01). Survival rate was significantly increased and apoptosis rate, TNF-α content, caspase 3 and 9 activities, and the expression of LOX-1 were significantly decreased in ox-LDL + RvE1 group compared to ox-LDL group (P < 0.01), these beneficial effects of RvE1 could be blocked by PI3K inhibitor wortmanin (P < 0.05).</p><p><b>CONCLUSION</b>The present data showed that RvE1 could effectively protect against ox-LDL-induced endothelial cell injury, which might be mediated by PI3K-AKT signaling pathway.</p>
Subject(s)
Humans , Apoptosis , Cells, Cultured , Eicosapentaenoic Acid , Pharmacology , Endothelial Cells , Metabolism , Lipoproteins, LDL , Signal Transduction , Umbilical Veins , Cell BiologyABSTRACT
Objective To study the effect of different levels of iodine nutrition on secretion of placental hormone in pregnant rats.Methods Two hundred and twenty five Wistar rats (165 female,60 male),weighing about 80 - 100 g were used in the study.Female rats were randomly divided into five groups according to their body weights:low iodine group Ⅰ(LⅠ),low iodine group Ⅱ (LⅡ),adequate iodine(control) group(Al),high iodine group Ⅰ ( HⅠ ),and high iodine group Ⅱ (H Ⅱ ),and 33 rats in each group.Animals in the low iodine groups were fed low-iodine diet,the iodine content was 13.46 μg/kg,in addition,these rats drank deionized water which containing potassium iodated,the dose was 0 and 5 μg/L,respectively.The rats of adequate and the two high iodine groups were fed normal diet,the iodine content was 22.00 μg/kg,they also drank deionized water,containing potassium iodated 50,3000,and 10000 μg/L,respectively.The rats mated after 3 months of feeding,and were respectively sacrificed at early pregnancy(5 ± 2)d,second trimester( 12 ± 2)d,and third trimester of pregnancy(17 ± 2)d,and then their serum was taken.Serum human chorionic gonadotropin(HCG),human chorionic thyrotropin(HCT),and progesterone were measured by enzyme-linked immunosorbent assay (ELISA).Results In the third trimester,the serum levels of rat HCG were significantly different between groups(F =4.16,P ＜ 0.05).The means of rats serum HCG of the two low iodine groups [ (16.08 ± 4.45),(17.43 ± 2.70)U/L] were significantly higher compared with that of AI group[ (13.68 ± 3.52)U/L] in the third trimester(all P ＜ 0.01 ).In the second and third trimester,the levels of rats serum HCT were significantly different between groups(F =3.59,3.40,all P ＜ 0.05).The means of rats serum HCT of HI group [(70.11 ± 10.97)μU/L] in the second trimester and HII group[(74.93 ± 13.22)μU/L] in the third trimester were higher than those of AI group[ (57.14 ± 12.56),(58.17 ± 8.54)μU/L] significantly(all P ＜ 0.01 ).There were statistical differences of the means of serum progesterone among trimester of pregnancy(F =4.06,4.43,all P ＜ 0.05).The level of serum progesterone of the third trimester[ ( 1462.80 ± 286.48 )pmoL/L] compared to those of the first[ (1929.93 ± 158.37) pmol/L] and the second trimester[ (1856.44 ± 542.08)pmol/L] was decreased significantly(all P ＜ 0.05) in LI group.In the control group,the level of serum progesterone of the second trimester [ (2046.45 ± 475.67)pmol/L ] was significantly higher than the first trimester[ (1714.39 ± 461.71 )pmol/L,P ＜ 0.05 ].Conclusions During pregnancy,placenta could promote HCG secretion under iodine-deficient conditions.In addition,the placenta increases the secretion of HCT under conditions of excess iodine.In the condition of severe iodine deficiency,the secretion of serum progesterone decreases,and further decreases with prolongation of pregnancy,but it is opposite to the change of HCG during pregnancy.This phenomenon could lead to harmful pregnant outcomes easily.
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<p><b>BACKGROUND</b>Lipoxins (LXs), endogenous anti-inflammatory and pro-resolving eicosanoids generated during various inflammatory conditions, have novel immunomodulatory properties. Because dendritic cells (DCs) play crucial roles in the initiation and maintenance of immune response, we determined whether LXs could modulate the maturation process of DCs and investigated the effects of lipoxin A(4) (LXA(4)) on lipopolysaccharide (LPS)-induced differentiation of RAW264.7 cells into dendritic-like cells.</p><p><b>METHODS</b>RAW264.7 cells were cultured in vitro with 1 microg/ml LPS in the absence or presence of LXA(4) for 24 hours, and cellular surface markers (MHC-II, CD80 (B7-1), CD86 (B7-2)) were measured by flow cytometry (FCM). Mixed lymphocyte reaction was performed to evaluate the allostimulatory activity. Cytoplastic IkappaB degradation and nuclear factor kappa B (NF-kappaB) translocation were detected by Western blotting. Luciferase reporter plasmid was transiently transfected into RAW264.7 cells, and luciferase activity was determined to measure the transcriptional activity of NF-kappaB.</p><p><b>RESULTS</b>LXA(4) reduced the ratio of LPS-treated RAW264.7 cells to DCs with morphological characteristics and inhibited the expression of MHC II. LPS-induced up-regulation of CD86 was moderately suppressed by LXA(4) but no obvious change of CD80 was observed. Moreover, LXA(4) weakened the allostimulatory activity of LPS-treated RAW264.7 cells. These alterations of LPS+LXA(4)-treated cells were associated with a marked inhibition of IkappaB degradation, NF-kappaB translocation and then the transcriptional activity of NF-kappaB.</p><p><b>CONCLUSIONS</b>LXA(4) negatively regulates LPS-induced differentiation of RAW264.7 cells into dendritic-like cells. This activity reveals an undescribed mechanism of LXA(4) to prevent excessive and sustained immune reaction by regulating maturation of DCs.</p>
Subject(s)
Animals , Mice , Biological Transport , Cell Differentiation , Cells, Cultured , Dendritic Cells , Cell Biology , I-kappa B Kinase , Metabolism , Lipopolysaccharides , Pharmacology , Lipoxins , Pharmacology , Macrophages , Cell Biology , NF-kappa B , Metabolism , Phenotype , Transcription, GeneticABSTRACT
The study is to investigate the effect of asiaticoside on collagen-induced arthritis (CIA). The model of CIA mice was prepared and the change of secondary paw swelling and the arthritis scores were observed. In vitro proliferation of spleen cells was examined using MTT assay. The cell-free protein extracts from the arthritic joints and nonarthritic joints were used for the analysis of protein expression of cyclooxygenase-2 (COX-2). And the level of PGE2 in joints was assayed using PGE2 express EIA kit. The tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) levels in the serum were measured by ELISA. Histopathological examination was performed by hematoxylin-eosin (HE) stain method. Asiaticoside (10, 20 and 40 mg x kg(-1) x d(-1), 22 d, ig) significantly reduced paw swelling, and decreased the arthritis scores. There was a significant reduction in proliferation of spleen cells of CIA mice treated with asiaticoside as compared with that of untreated CIA mice. COX-2, PGE2, TNF-alpha and IL-6 production in CIA mice were inhibited by asiaticoside. Meanwhile, the pathological examination showed that articular cartilage degeneration with synovial hyperplasia and inflammatory cells infiltration in CIA mice was suppressed by asiaticoside. Its active mechanism may be related to inhibiting proliferation of lymphocyte and reduction of expression of COX-2 and inflammatory cytokines.
Subject(s)
Animals , Male , Mice , Ankle Joint , Metabolism , Pathology , Anti-Inflammatory Agents , Pharmacology , Arthritis, Experimental , Metabolism , Pathology , Cell Proliferation , Centella , Chemistry , Collagen Type II , Cyclooxygenase 2 , Metabolism , Cytokines , Blood , Metabolism , Dinoprostone , Metabolism , Interleukin-6 , Blood , Lymphocytes , Pathology , Mice, Inbred DBA , Plants, Medicinal , Chemistry , Spleen , Pathology , Triterpenes , Pharmacology , Tumor Necrosis Factor-alpha , BloodABSTRACT
Objective cyclin D1 gene plays a significant role in regulating cell cycle progression.Suppression of cyclin D1 protcin expression can effect on cellular proliferation,distribution of cell cycle and apoptosis.This study was to determine whether this effect also existed in chronic leukemia ceil line K562 by inhibiting the expression of cyclin D1 protein through RNA interference in vetro.Methods Plasmid vectors expressing small hairpin RNA (shRNA) targeting at cyclin D1 gene were constructed and transfected into K562 cells by chitosan,cyclin D1 protein was examined by using Western blot analysis.Inhibition of cellular proliferation was evaluated hy soft agar colony formation assay.The cell cycle and apoptosis were determined by flow cytometry.Results Expression of cyclin D1 protein was markedly down-regulated and capability of colony formation was suppressed after transfection with pshRNA-419 and pshRNA-575 at 48h.Down-regulation of cyclin D1 protein could effect on distribution of cell cycle arrested at G_0/G_1 phase and markedly induce apoptosis of K562 cells.But there had no above biological effects ob- served after transfection with blank vector and control vector of m-pshRNA-790.Conclusion Down-regulation of cyclin D1 expression can inhibit growth of K562 cells,and effect on distribution of cell cycle arrested at G_0/G_1 phase.The primary results suggest that cyclin D1 gene might serve as an effective target for the treatment of leukemia.
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<p><b>OBJECTIVE</b>To observe the effect of tetramethylpyrazine (TMP) on lipopolysaccharides (LPS) induced macrophage cyclo-oxidase-2 (COX-2) gene expression and activity in RAW264.7 mice, and to further investigate the effect and mechanism of TMP on LPS induced apoptosis of cardiac myocytes in suckling mice.</p><p><b>METHODS</b>RT-PCR and Western Blot (WB) were used to investigate the macrophage COX-2 gene expression, ELISA was used to measure its activity, fluorescence microscopy was used to determine the apoptosis of murine neonatal cardiac myocyte, and fluorescence spectrophotometry was used to detect the concentration of intracellular calcium ion (Ca2+).</p><p><b>RESULTS</b>TMP of 10(-6) mol/L could significantly reduce the COX-2 mRNA and protein expression (P < 0.05), in 10(-5) mol/L and 10(-4) mol/L could significantly decrease the COX-2 expression (P < 0.01) stimulated by LPS, but couldn't influence the activity of COX-2 by different TMP concentration. TMP in 10(-5) mol/L could significantly lower the concentration of intracellular Ca2+ in cardiac myocyte, and antagonize the LPS induced apoptosis of cardiac myocyte in suckling mice (P < 0.05).</p><p><b>CONCLUSION</b>TMP has the pharmacological effect in inhibiting LPS induced macrophage COX-2 expression and apoptosis of cardiac myocyte in suckling mice.</p>