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ObjectiveTo investigate the relationship between tumor necrosis factor (TNF) α308,TNF-β252 genotypes and serum TNF-α and TNF-β levels in patients with gastric cancer (GC).MethodsA total of 57 pathological diagnosed GC patients were collected,of which 49 cases were from Zhongnan Hospital of Wuhan University and 8 cases were from Tumor Hospital of Hubei Province.Another 18 age and sex matched healthy controls were enrolled at the same time.The TNF-α308 and TNF -β252 polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).The serum TNF-α and TNF-β levels of 57 GC patients and 18 healthy controls were measured by ELISA.The difference of TNF serum level in different TNF genotypes of GC and the difference between GC patients and the healthy controls was analyzed.Its relationship with clinical pathological characters was also analyzed.Results With TNF-α308 genotype,GA were 6 cases and GG were 51 cases.With TNF-β252 genotype,GG were 17 cases,GA and AA each were 20 cases.The serum TNF-α level of GC patients was significant higher than that of healthy controls (median 445 × 10-3 μg/L vs 5 × 10-3 μg/L,P＜0.05),and the serum level of each TNF-α308 andTNF-β 252 genotypes was significant higher than that of healthy controls (P＜0.05).However there was no statistical significance in TNF -β level compared with healthy controls (P＞0.05).In addition,the serum TNF-α levels of the TNF-α308G/TNF-β252G and TNF-α308G/TNF β252A haplotypes in GC patients were significant higher than those of the healthy controls (P＜0.05),and the increase like serum TNF-α level was associated with the patients'age and lymph node metastasis (P＜0.05).The TNF-β level in patients with TNF-α308A and TNF-β252G high-risk haplotypes showed a significant relation with smoking history (P＜ 0.05). Conclusions Serum TNF-α level of GC patients was significantly higher,however there was no significant association between the increase and TNF-α308 and TNF-β252 genotypes.The serum TNF-α levels of TNF-α308G/TNF-β252G and TNF-α308G/TNF-β252A haplotypes in GC patients were significant higher,and associated with the patients'age and lympb node metastasis.It was indicated that TNF haplotypes may have certain impact on the TNF expression and clinical subtypes in GC.
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Objective To investigate the association of the major histocompatibility complex class Ⅰ chain-related antigens A (MICA)-129 gene polymorphism and soluble MICA (sMICA) levels with ulcerative colitis (UC) in Hubei Han nationality. Methods The genetic polymorphism of MICA-129 was examined using a polymerase chain reaction-sequence based test (PCR-SBT) in 256 UC patients and 460 healthy controls. From the above subjects, 80 patients and 90 healthy individuals were randomly selected for determining serum sMICA concentrations by ELISA. Results The frequencies of variant allele (G) and genotype (GG) in MICA-129 gene were significantly higher in the UC patients than in the controls(76. 8%vs 72. 2%, P =0. 060; 55.9% vs 46. 3% ,P =0. 016). Serum sMICA levels were significantly elevated in the patients compared to the controls[(576. 47 ±279. 02) ng/L vs( 182. 17 ±73. 11 ) ng/L,P ＜0. 001]. In addition, the sMICA levels were higher in the patients carrying MICA-129 GG genotypes than in those carrying ( GA + AA) genotypes [( 638. 87 ± 347. 15 ) ng/L vs ( 507. 51 ± 152. 87 ) ng/L, P = 0. 035].Conclusions The genetic polymorphism of MICA-129 and sMICA levels are correlated with the UC patients in Hubei Han nationality. Our findings demonstrate that MICA-129 gene may contribute to the pathogenesis of UC.
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Objectives The present study aimed to investigate the associations between genetic polymorphism of methylenetetrahydrofolate reductase ( MTHFR) G1793A, plasma homocysteine (Hcy) levels, vitamin status and ulcerative colitis ( UC) in a cohort of patients in Hubei Han nationality. Methods Two hundred and ninty-nine UC patients and 764 age- and sex-matched healthy controls were recruited in this study. Polymorphism of MTHFR G1793A was examined using a PCR-RELP method.Plasma levels of Hcy, folate and vitamin B12 were determined by enzymatic cycling assay and corpuscle immune chemiluminescence assay, respectively.Results Both variant allele and genotype frequencies in MTHFR G1793A gene were significantly higher in the UC patients compared to the controls (22.24% vs 14.20% , P<0.001 ;42.81% vs 26.97%, P < 0.001, respectively).Plasma Hcy levels were increased in UC patients compared to the controls [(20.67 ±6.42)mmol/L vs (13.21 ±5.11)mmol/L, P <0.001] while folate and vitamin B12 concentrations were significantly decreased [(11.37±6.34) nmol/L vs (14.89±7.21) nmol/L, P < 0.001; (324.15±127.53 ) pmol/L vs (421.54±128.45 ) pmol/L, P < 0.001, respectively].Furthermore, hyperhomocysteinaemia (HHcy) and folate deficiency were also more prevalent in the UC patients (32.44% vs 25.78% , P = 0.029; 23.41% vs 17.01%, P =0.016, respectively).Conclusions Genetic polymorphism of MTHFR G1793A Wag strongly associated with UC.HHcy,folate deficiency and low vitamin B12 concentration were common phenomena in the UC patients of Hubei Han nationality.Our findings demonstrate that the genes relmed to Hey metabolism may play an important role in the pathogenesis of UC.
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Objective To evaluate association of plasma levels of homocysteine, folate and vitamin B12 as well as genetic polymorphisms of homocysteine with ulcerative colitis (UC). Methods Three hundred and ten consecutive patients with UC and 936 healthy controls were recruited.Polymorphisms of methylenetetrahyrdofolate reductase (MTHFR, C677T and A1298C), methionine synthase (MTR) A2756G and methionine synthase reductase (MTRR) A66G were genotyped using PCR-RELP methods. Eighty eight patients and one hundred healthy controls were randomly selected for determination of plasma levels of homocysteine by enzymatic cycling assay, and concentrations of folate and vitamin B12 were measured by corpuscle immune chemiluminescence assay. Results The variant allele and genotype frequencies of MTHFR 1298C, MTR 2756G and MTRR 66G were significantly higher in UC patients than in the healthy controls (P＜0. 01). Moreover, plasma homocysteine level was obviously higher in UC patients than in controls [(21.73±6.59) mmol/L vs(12.47±5.01)mmol/L,P<0.01).Whereas both folate F(11.25±6.19)nmol/L] and vitamin B12 [(322.81±128.47)pmol/L] concentrations were significantly lower in UC patients than in controls [(15.28±7.72)nmol/L and (422.59±129.36)pmol/L,respectively,P<0.01].Logistic analysis revealed that abnormal levels of homocysteine,folate and vitamin B12 were independent risk factors for UC(P<0.01).Conclusions Plasma levels of homocysteine,folate and vitamin B12 as well as the related genetic polymorphisms of homocystein are correlated with UC,which provides a theoretical basis for supplement of folate and vitamin B12 in treatment of UC patients.
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Aim To study the effect of PLC on rabbit and human platelet actin polymerization, and then to explore the mechanism of PLC anti-aggregation to platelet. Methods Platelets of rabbit and human were treated with PSS, ASA and different doses of PLC respectively and then were extracted by Triton abstraction. The relative concentration of actin of differently treated platelets induced by ADP was determined by SDS-PAGE and spectrophotometre. Results For rabbit platelets were treated with PSS, the relative concentration of actin determined at static state was 1.682?0.319; when the platelets were treated with ASA 668 ?mol?L -1,PLC 5,10,15,20 and 25 U?ml -1, the relative concentration of actin determined at activated state induced by ADP was 2.450?0.562,1.089?0.322,1.727?0.442,1.450?0.324,1.161?0.306, 0.857?0.242 and 0.692?0.187 respectively. Compared with PSS, inhibition rates (%) of ASA 668 ?mol?L -1, PLC 5, 10, 15, 20, 25 U?ml -1 to the relative concentration of actin were 55.55,29.51, 40.82,52.61, 65.02,71.76 respectively.For human platelets were treated with PSS, the relative concentration of actin determined at static state was 1.358?0.376; when the platelets were treated with ASA 668 ?mol?L -1,PLC 5,10,15,20 and 25 U?ml -1, the relative concentration of actin determined at activated state induced by ADP was 2.445?0.750, 1.096?0.344, 1.705?0.507,1.437?0.416, 1.165?0.355, 0.845?0.257 and 0.679?0.198 respectively. Compared with PSS, inhibition rates (%) of ASA 668 ?mol?L -1, PLC 5, 10, 15, 20, 25 U?ml -1 to the relative concentration of actin were 55.17,30.27, 41.23,52.35, 65.44, 72.23 respectively. Conclusion PLC has significant effects on actin polymerization of rabbit and healthy human platelets (P
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AIM To investigate the antiplatelet effect of PLC by de termining the concentration of TXB 2, 6-keto-PGF 1? and bleeding time( BT). METHODS TXB 2 and 6-keto-PGF 1? were detected by ra dioimmunity kit. Bleeding time were measured by routine methods. RESULTS Six doses of PLC can prolong BT significantly(P0 05). The inhibition on TXB 2 generation, of PLC 60 0～ 1 000 U?kg -1 with ADP revulsant, PLC 800～1000 U?kg -1 with AA revulsant and PLC 1 000 U?kg -1 with Collagen revulsant, is more significant than ASA (P
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AIM To investigate the antiplatelet effects of phospholipase C (PLC) by studying the effects of PLC on platelet adhesion and aggregation. METHODS Auxiliary agent(-), aspirin (+) and six doses of PLC were administered to anaesthetized rabbit via duodenum. Blood was taken from artery carotis before administration and at 1,2,4 hours after administration. Platelet aggregation rates were determined by turbidmetry. Platelet adhesion rates were tested by glass ball method. RESULTS 100 IUPLC?L -1 and 200 IUPLC?L -1 had no evident effect on rabbit's platelet adhesion rate, 400～ 1 000 IUPLC?L -1 decreased the platelet adhesion rate significantly ( P
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AIM To investigate the effects of PLC on ultrastructure of platelets. METHODS The effects of PLC on ADP induced platelet aggre-gation were detected by tubidmetry; the ultrastru-cture changes of platelet were analyzed by electron microscope. RESULTS The rate of PLC inhibited significantly platelet aggregation by ADP induced is 86 03%?12 06% and 82 47%?5 49%. Pseudopodia can inhibit by 0 5 U PLC, at this group increase in the number of the granules associated with enhanced intensities of their electron densities and smaller dilated canalicular system. compared with normal saline group. In the 2 5 U PLC group the platelet morphology and ultrastructure approach blank group. The platelet is very smooth. PLC can inhibit producing pseudopodia-like process and the releasing of granules. Dilated canalicular system is similar to blank group, But it was different to normal group. CONCLUSION It's showed that PLC can significantly inhibit platelet aggregation and ultrastructural changes.
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AIM Aim to study the effect of PLC on human platelet cytoplasmic free calcium concentration, and then to explore the mechanism of PLC anti aggregation to platelet. METHODS Washed platelet of human was marked with fluorescence indicator Furu 2/AM, and then was treated with PSS, ASA and different dose of PLC respectively. Cytoplasmic free [Ca 2+ ] i of differently treated platelet at static state and activated sate induced by ADP was determined by double beam fluorescence spectrophotometric method. RESULTS Healthy human platelet was treated with PSS, ASA 334 ?mol?L -1 , 2 5, 3 75, 5, 10 and 20U PLC?ml -1 . Cytoplasmic [Ca 2+ ] i determined at static state was 152 55?15 07, 131 63?15 58, 140 27?12 03, 139 48?1 73, 121 11?9 58, 116 62?15 96 and 107 20?17 07 respectively. [Ca 2+ ] i determined at activated sate induced by ADP was 902 62?94 74, 687 99?62 86, 810 99?72 37, 701 73?21 37, 429 67?71 59, 342 82?44 86 and 263 27?25 46 respectively. Compared with PSS, inhibition rates ( % ) of ASA 334 ?mol?L -1 , 2 5, 3 75, 5,10, 20 U PLC?ml -1 to the increment of activated platelet cytoplasmic [Ca 2+ ] i were 25 83, 10 58, 25 04, 58 86, 69 84, 79 19 respectively. At the same condition, inhibition rate ( % ) of 10 U PLC?ml -1 to the increment of five hypertensive activated platelet cytoplasmic [Ca 2+ ] i were 74 25, 73 48, 61 32, 82 87,70 60 respectively. CONCLUSION PLC has significant effects on static cytoplasmic[Ca 2+ ] i of healthy and hypertensive human platelet ( P