Objective To investigate the relationship between the microsatellite polymorphism of cytotoxic T lymphocyte associated antigen-4 (CTLA-4) gene and inflammatory bowel disease(IBD) patients in Zhejiang province. Methods One hundred and eighteen IBD patients without consanguinity (99 patients with ulcerative colitis, 19 patients with Crohn's disease) and 140 healthy controls were studied. The (AT) repeats in the 3' untranslated region of exon 4 of CTLA-4 gene were amplified by allele-specific PCR. The amplified products were electrophoresed with 12% polyacrylamide gel, followed by silver staining. Results Twenty alleles were found in IBD patients and healthy controls. The frequency of 122 bp allele was higher in patients with ulcerative colitis (P=0.0001/Pc=0.0025, OR= 11.393, 95% CI:2.574-50.429) and Crohn's disease (P=0.0003/Pc=0.0050, OR=21.061, 95% CI:3.927-112.94) than that in healthy controls. Conclusion CTLA-4 gene microsatellite polymorphism was obviously associated with IBD in Zhejiang province.

Adult ; Agriculture ; Antibody Formation ; Female ; Humans ; Immunity, Cellular ; Male ; Middle Aged ; Occupational Exposure

Objective To study the distribution of IL-10 and TNF gene polymorphisms in patients with gastroduodenal diseases in Hubei Han ethnic and their association with Helicobacter pylori (Hp) infection. Methods Six hundred and five patients with gastroduedenal diseases (196 chronic gastritis, 189 gastroduodenal ulcer and 220 gastric cancer) as well as 624 healthy controls were genotyped with PCR-RFLP method for IL-10-1082,-819,-592 and TNFα-308, lymphotoxin-α (LTα) Nco Ⅰ and AspH Ⅰ gene polymorphisms. Hp infection status was determined with ELLS& Results (1) There was significant difference of IL-10-1082 AG + GG genotype among the gastric cancer group with the non-malignant gastric diseases groups and healthy control group (P <0. 05). There was no significant difference of IL-10-592 and -819 gene polymorphisms among gastric cancer patients,non-malignant gastric disease patients and healthy controls (P>0. 05). The genotype frequencies of IL-10-819 were the same as those of IL-10-592. (2) Frequency of IL-10-1082 AG + GG genotype in gastric cancer patients with positive Hp was significantly higher than that in the other three groups (P < 0. 05). (3) Frequency of LTα Nco I AG genotype in gastric cancer patients with Hp infection was signiilcandy higher than that in Hp positive healthy controls (P < 0. 05). There were no other associations between TNFα-308, LTα Nco Ⅰ and AspH Ⅰ gene polymorphisms and Hp infection in gastroduodenal diseases. Conclusions (1) Allele AG + GG of IL-10-1082 was associated with gastric cancer in Han nationality of Hubei province. (2) IL-10-1082 AG + GG,LTct Nco ⅠAG heterozygous genotype may be associated with Hp infection in patients with gastric cancer in Han nationality of Hubei province.

Salvia miltiorrhiza is a traditional Chinese medicine (TCM) and is widely used as a clinically medication for its efficiency in treating cardiovascular disease. Due to TCM is a comprehensive system, the mechanism of S. miltiorrhiza treating cardiovascular disease through integrated multiple pathways are still unclear in some aspects. With the rapid progress of bioinformatics and systems biology, network pharmacology is considered as a promising approach toward reveal the underlying complex relationship between an herb and the disease. In order to discover the mechanism of S. miltiorrhiza treating cardiovascular disease systematically, we use the auxiliary mechanism elucidation system for Chinese medicine, built up a molecule interaction network on the active component targets of S. miltiorrhiza and the therapeutic targets of cardiovascular disease to offer an opportunity for deep understanding the mechanism of S. miltiorrhiza treating cardiovascular disease from the perspective of network pharmacology. The results showed that S. miltiorrhiza treating cardiovascular disease through ten pathways as follows: improve lipid metabolism, anti-inflammation, regulate blood pressure, negatively regulates blood coagulation factor and antithrombotic, regulate cell proliferation, anti-stress injury, promoting angiogenesis, inhibited apoptosis, adjust vascular systolic and diastolic, promoting wound repair. The results of this paper provide theoretical guidance for the development of new drugs to treat cardiovascular disease and the discovery of new drugs through component compatibility.
Animals ; Cardiovascular Diseases ; drug therapy ; genetics ; metabolism ; Databases, Factual ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Gene Regulatory Networks ; drug effects ; Humans ; Salvia miltiorrhiza ; chemistry ; Signal Transduction

OBJECTIVETo establish the steriod-induced osteoporosis model with the type of Kidney-Yin deficiency.

METHODSTotally 45 female Kunming mice were randomly divided into normal group,model group and Liuwei Dihuang pills(Chinese character: see text)group. The model was established by intramuscular injecting of Dexamethasone. Liuwei Dihuang pills (Chinese character: see text) group was administered orally with Liuwei Dihuang pills (Chinese character: see text). The signs and symptoms of mice were observed dynamically. All the animals were sacrificed at the end of the 6th weeks. The level of ACTH, cAMP, cGMP, TSH and E2 in serum were detected to evaluate deficiency of Kidney-Yin. Morphological changes and bone density were observed to evaluate osteoporosis.

RESULTS(1) Compared with control group, mice in model group appeared obvious Kidney-Yin deficiency symptoms, including hair dry, restlessness, excitability, hard stool, and yellow. (2) Compared with control group,the weight of mice in model group gained slower (P<0.01); the index of adrenal gland,liver and spleen decreased (P<0.01, P<0.01 ,P<0.01); the level of ACTH and TSH increased (P<0.01 ,P<0.01), the level of E2 decreased (P<0.01) and the ratio of cAMP/cGMP increased (P< 0.05). (3)Compared with control group,the bone density of lumbar vertebra and femur in model group were significantly decreased (P<0.01, P<0.05); HE staining revealed osteoporosis in model group mice. (4)However, the Liuwei Dihuang pills (Chinese character: see text) group can partly antagonize the inhibition of the HPA axis, alter the disordered sex hormone and the ratio of cAMP/cGMP, and reverse the osteoporosis partly.

CONCLUSIONthe model of osteoporosis with type of Kidney-Yin deficiency could be established by Dexamethasone intramuscular injection. With less interference, it wight be a stable and reliable modeling method for integration of disease and syndrome in TCM.

Animals ; Bone Density ; Dexamethasone ; toxicity ; Disease Models, Animal ; Female ; Kidney Diseases ; etiology ; Medicine, Chinese Traditional ; Mice ; Osteoporosis ; chemically induced ; Yin Deficiency ; complications

Objective To investigate the expression of serum soluble cytotoxic T lymphocyte associated antigen 4 (sCTLA4), the association of sCTLA4 level with erythrocyte sedimentation rate (ESR) and C reactive protein (CRP), as well as its role in patients with Crohn's disease (CD). The relationship-1661A/G and -1722T/C polymorphisms of CTLA4 gene and between disease susceptibility and phenotype of CD was analyzed. Methods A total of 126 CD patients and 300 healthy controls were enrolled in the study. Serum sCTLA4 level was determined by enzyme-linked immunosorbent assay. The concentrations of ESR and CRP were analyzed by automatic ESR Analyzer SRS 100/Ⅱ and rate nephelometry, respectively. The polymorphisms of CTLA4-1661A/G and -1722 T/C were genotyped by DNA sequencing. Results Serum sCTLA4 level was higher in CD patients than in healthy controls [(18. 70±3. 72) ng/ml vs (1.72±0. 32) ng/ml, P＜0. 01)]. Among CD patients, sCTLA4 level was higher in patients with active disease when compared to those with inactive disease [(19.83±4.35) ng/ml vs (18. 02±3.14) ng/ml, P=0. 015)]. sCTLA4 level was positively correlated with ESR and CRP levels (r=0. 267, P=0. 003; r=0. 524 P ＜0.01, respectively). In CD patients, serum sCTLA4 level was significantly higher in those with stricturing disease behavior than that in those without stricturing and penetrating or with penetrating disease behavior (P= 0.021; P=0. 015, respectively). Detection of CTLA4 -1661A/G and -1722T/C polymorphisms showed no significant difference between CD patients and healthy controls. Conclusions The high level of serum sCTLA4 in CD patients is correlated with disease activity, CRP levels and disease behavior. It suggests that sCTLA4 may play an important role in pathogenesis of CD.

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.

Objective To investigate the correlation between genetic polymorphisms of glutathione S-transferases (GSTs) and ulcerative colitis (UC) in Hubei Han population. Methods Genetic polymorphisms of GSTM1 and GSTT1 of 270 patients with UC (UC group) who were admitted to the Zhongnan Hospital, People's Hospital of Wuhan University, Tongji Hospital and Union Hospital of Huazhong University of Science and Technology from August 2002 to December 2009 and 623 healthy people ( control group) were detected by restriction fragment length polymorphism-polymerase chain reaction. All UC patients were allocated to distal UC group (n= 229) and extensive UC group (n =41 ) according to the location of the lesions; and all UC patients were also allocated to mild-moderate group (n = 237) and severe group (n = 33 ). The genetic polymorphisms of GSTP1 of these patients and healthy people were detected by polymerase chain reaction. The genotypes of GSTM1, GSTT1 and GSTP1 were also detected. GSTM1 and GSTT1 containing small DNA segments ( 157 bp and 480 bp) were defined as GSTM1 (+) and GSTT1 (+), otherwise, GSTM(-) and GSTT1 (-), respectively. All data were analyzed by chisquare test. Results The frequencies of GSTM1(-), GSTT1(-) and GSTP1 (Val/Val) were 70.7% (191/270),64.8% (175/270) and 48.9% (132/270) in the UC group, and 41.7% (260/623), 47.2% ( 294/623 ) and 34.3% (214/623) in the control group, with a significant difference between the two groups (x2 = 63. 404,22. 320, 25. 384, P ＜0.05 ). The frequencies of GSTT1 (-) and GSTP1 (Val/Val) were 71.6% (164/229) and 57.6% (132/229) in the distal UC group, which were significantly higher than 31.7% (13/41) and 29.3%( 12/41 ) in the extensive UC group ( x2 = 24.528, 9.609, P ＜ 0.05 ). The frequencies of GSTM1 (-) were 65.1%(149/229) in the distal UC group and 56.1% (23/41) in the extensive UC group, with no significant difference between the two groups ( x2 = 1. 210, P ＞ 0.05 ). The frequencies of GSTT1 (-) and GSTP1 ( Val/Val ) were 71.6%(164/229), 31.7% ( 13/41 ) in the distal UC group and 57.6% ( 132/229), 29.3% ( 12/41 ) in the extensive UC group, with a significant difference between the two groups ( x2 = 24. 528, 9. 609, P ＜ 0. 05 ). There was no significant difference in the frequencies of GSTM1 (-), GSTT1 (-), GSTP1 (Val/Val) in the mild-moderate group and the severe group( x2 = 0. 623, 1. 884, 3. 403, P ＞ 0. 05 ). Conclusions Variant genotypes of GSTs are significantly correlated with UC in Hubei Han population. The severity of UC may not be correlated with variant genotypes of GSTs.

ObjectiveTo make ultrasound anatomy teaching samples of odynopoeia fetus heart with complex abnormity consistent with echocardiographic view,and utilize them in echo teaching.MethodsTen odynopoeia foetus hearts were cutted comparing to different echocardiographic imaging respectively.Results Dissections of fetus heart were obtained including 5 cases of single cardiac ventricle(2 case with interruption of aortic arch),4 cases of double outlet of right ventricular,and 1 case of hypoplastic left heart syndrome with serious coarctation of the aorta.The apical long axis view,the three blood vessel view,as well as the long axis view of the aortic arch were typical views of ultrasound diagnosis and teaching.ConclusionsIt is profit to raise and accurate rate of fetus echocardiography diagnosis by studying dissection of fetus heart anatomic structure with complex abnormity,which would make ultrasound imaging visualization.

Objective To evaluate the accuracy, of different threshold segmentation of 18FDG PET for target volume delineation of non-small cell lung cancer(NSCLC) and the potential influence on radiotherapy treatment planning. Methods Eight NSCLC patients who had tumor with clear margin on CT scan and the amplitude of tumor movements not more than 5 mm were enrolled. PET scans were carried out at 1 h after intravenous injection of 18FDG with CT image for attenuation revisement. Gross target volume (GTV) delineated on CT image ( GTVCT ) was used as the standard. Then, GTVs were delineated on PET image with three different threshold segmentation of 42% Imax(total) (42% of maximum voxel intensity within the tumor) ,Iback + 20% Imax-back(max) (mean background intensity + 20% of normalized background-subtracted maximum voxel intensity within the tumor) and Iback -20% Imax-back(slice) (mean background intensity + 20% of normalized background-subtracted maximum voxel intensity of each slice within the tumor) ,the corresponding GTV was named as GTV42%, GTV20%max and GTV20%slice. Both the size of GTV42%, GTV20%max, GTV20%slice and GTVCT,and the coverage over GTVCT for each GTV were compared. A three dimensional margin of 1 cm were added to GTVCT, GTV42%, GTV20%max and GTV20%slice to form corresponding PTVCT, PTV42%, PTV20%max and PTV20%slic e. Three dimensional conformal radiotherapy treatment plans were designed based on PTVCT,PTV42% , PTV20%max and PTV20%slice respectively for each patient. The prescription dose of all PTVs was 66 Gy in 33 fractions in 6.6 weeks. Both the volume accepting dose less than 95% of prescription dose within PTVCT ( VPTV ) and the lung V20 were compared among the four plans based on different PTVs. Tumor control probability(TCP) as well as lung normal tissue complication probability (NTCP) were also compared. Resuits Eight patients were enrolled in this study. Median deviation of volume between GTVPET and GTVCT were -54.1% , -21.5 % and 5.3 % for GTV42% , GTV20%max and GTV20%slice, respectively. Median coverage over GTVCT of GTV42% , GTV20%max and GTV20%slice was 45.9% ,78.0% and 95.3% respectively( F = 57.50,P＜0.01). Median 7.5% of VPTV was observed for radiotherapy treatment plan based on PTV42% ,which meant that it might induce median 1% decrease of TCP comparing with that of radiotherapy treatment plan based on PTVCT. Whereas,there were only 1.3% and 0.0% of VPTV for treatment plans based on PTV20%max and PTV20%slice respectively. As far as TCP was concemed, both PTV20%max group and PTV20%slice group were superior to PTV42% group,there was no significant difference among PTV20%max group, PTV20%slice group and PTVCT group. Lung V20 and lung NTCP showed no significant difference among all groups. Conclusions The threshold segmentation of Iback + 20% Imax-back(slice) , being slice specialized, might be an optimal threshold segmentation for target volume delineation of lung caner. Independent of information of target volume provided by CT scan in advance,it is recommended to use for the target volume delineation of NSCLC with atelectasis.