Adult ; Arterio-Arterial Fistula ; pathology ; Coronary Vessels ; pathology ; Female ; Humans ; Pulmonary Artery ; pathology

OBJECTlVE To investigate the inhibition of Radix lsatidis and its major constituents indigo and indirubin on three principal subtypes of organic anion transporters ( OATs) , Oat1, Oat2 and Oat3 in vivo in mice. METHODS Granules of Radix lsatidis ( GRl) 0.615 and 2.46 g·kg-1 , decoction of Radix lsatidis ( DRl) 1.6 and 6.4 g·kg-1 , indigo 0.008 and 0.64 mg·kg-1 and indirubin 0.0192 and 1.536 mg·kg-1 were ig given to the NlH mice (60 mice per group), twice a day, for 5 d while four control groups were set up, including vehicle of water, 0.5%sodium carboxymethyl cellulose ( CMC) , positive control probe-necid (0.05 g·kg-1) and additives of sucrose plus dextrin (1.5 g·kg-1 each) groups. After the last dosing of the test samples, para-aminohippuric acid ( PAH) clearance test was conducted. All the mice were iv given PAH 0.03 g·kg-1 and 1, 2.5, 5, 7.5, 10 and 20 min later before 10 mice per group were euthanized to collect whole blood and the kidneys were quickly removed. Each right kidney was homoge-nized to analyze the PAH accumulations and each left kidney to extract total mRNA for analysis of Oat1, Oat2 and Oat3 gene expressions using quantitative real-time PCR. The concentrations of PAH in sera and in kidney homogenates were determined by the method of Kiguchi. Major pharmacokinetic parame-ters of PAH in sera were calculated by pharmacokinetic software ( DAS2.0) . PAH uptake test for kidney slices was performed on another group of NlH mice according to the method of Nakakariya. RESULTS There was no significant difference between water control group and 0.5%CMC group in all the examined items. Compared with the vehicle control groups ( water and 0. 5%CMC group ) , elimination half time ( t1/2β) of PAH in GRl 2.46 g·kg-1 ,indigo 0.64 mg·kg-1 and indirubin 1.536 mg·kg-1 groups was signifi-cantly prolonged (P<0.05), the total clearance (Cl) and volume of distribution (Vd) were obviously reduced ( P<0.01) and the area under the curve ( AUC0-20 min ) of PAH in all the tested groups was signifi-cantly increased ( P<0.01) . AUC0-20 min obtained from renal PAH accumulations within the checked time was significantly higher ( P<0.05, P<0.01) than in the vehicle control group. But there was in no signifi-cant difference between all the study groups in kidney-to-plasma AUC ratios. PAH uptake results by kidney slices were significantly lower ( P<0. 05, P<0. 01 ) than in vehicle control group in every two dosages of all the four samples tested. Compared with vehicle control group, the mRNA expressions of Oat1, Oat2 and Oat3 were obviously ( P<0.05, P<0.01) and abnormally regulated in the groups of GRl 2.46 g·kg-1, DRl 6.4 g·kg-1, indigo 0.64 mg·kg-1 and indirubin 1.536 mg·kg-1. CONCLUSlON The renal Oat1, Oat2 and Oat3 of mice are significantly inhibited by GRl, DRl, indigo and indirubin. The inhibitory function of Radix lsatidis probably stems from indigo and indirubin contained in it.

Objective To investigate the effect and mechanism of anti-tumor necrosis factor (TNF)-α on the intestinal mucosal permeability in dextran sulfate sodium (DSS) induced colitis mice.Methods Eighteen C57BL/6J mice were evenly divided into healthy control group,model group and anti-TNF-α treated group.The mice of model group and anti-TNF-α treated group were fed with 5％DSS solution for 7 days.The mice of anti-TNF-α treated group were injected anti-TNF-α (5 mg/kg)intraperitoneally on the first and fourth day; control group and model group were substituted with equal volume saline injection.The mice were sacrificed at 7 days after modeling.The disease activity index (DAI) score was evaluated everyday.The intestinal permeability was examined by Evan′s blue (EB) method and FITC-dextran (FITC-D) method.The colon tissue was collected for observation under microscope and histological index (HI).The small intestinal tissues were examined under electron microscope.The 10％ homogenate of colon and intestinal mucosa was prepared,the activity of myeloperoxidase (MPO),the content of TNF-α and epithelial myosin light chain kinase (MLCK) concentration were determined with kits respectively.The expression of MLCK in intestinal mucosa was tested by Western blot assay.Single factor of variance between groups were analyzed.Results Compared with control group,the DAI of model group increased daily.Compared with model group,the DAI of anti-TNF-α treated group improved.In model group,mice intestinal epithelial cells junctional complex shortened and widened and the cell gap expanded.In anti-TNF-α treated group,the connection structure of mice intestinal epithelial cells was tighter.The activity of HI and MPO and the content of TNF-α of model group were higher than those of control group (P ＝ 0.008,0.006 and 0.001,respectively),all of those of anti-TNF-α treated group were lower than those of model group (P=0.004,0.008 and 0.005,respectively).The F value of three groups was 131.98,218.28 and 58.93,respectively.The contents of EB in mice intestinal wall and serum FITC-D of model group were higher than those of control group (P=0.003 and 0.010),and those of anti-TNF-α treated group were lower model group (P＝0.001 and 0.009).The F value of three groups was 69.36 and 17.96.The MLCK concentration in mice intestinal mucosa of model group [(71.10± 7.52) ng/g] was higher than that of control group [(18.56±9.92) ng/g,P＜0.01],that of anti-TNF-α treated group [(37.56±15.84) ng/g] was lower than model group (P=0.008),and the difference among these three groups was statistically significant (F＝ 17.23).The Western blot results indicated the expression of MLCK in intestinal mucosa of model group was higher than that of control group,and that of anti-TNF-α treated group was lower than model group.Conclusions Anti-TNF-α play an important role in improving colitis,and the intestinal mucosal permeability.The mechanism may be related with the regulation of MLCK expression.

Background:CpG island methylator phenotype(CIMP)involving tumor suppressor gene( TSG)on short arm of chromosome 3(chromosome 3p)has been found in various types of cancers. However,its correlation with gastric cancer has not been clarified. Aims:To study the clinical significance of CIMP involving TSG on chromosome 3p in gastric cancer. Methods:Methylation specific PCR(MSP)was used to examine methylation profiles for hOGG1,VHL,RAR-B, hMLH1,SEMA3B,RASSF1A,BLU and FHIT harbored in chromosome 3p in 100 gastric cancer and paired paracancerous tissues. High CIMP( CIMP-H)was referred for those samples having four or more synchronously methylated genes. Relationship between CIMP-H and clinicopathological characteristics in gastric cancer was analyzed. Results:Positive methylation rates of VHL(P = 0. 030),hMLH1(P < 0. 001),SEMA3B(P = 0. 003),RASSF1A(P < 0. 001)and FHIT(P < 0. 001)were significantly higher in gastric cancer tissue than in paracancerous tissue. Incidence of CIMP-H rate in gastric cancer tissue was significantly higher than that in paracancerous tissue(44. 0% vs. 4. 0% ,P < 0. 001). CIMP-H was negatively correlated with degree of tumor differentiation(P = 0. 004),and positively correlated with lymph node metastasis(P = 0. 005),but not correlated with gender,age,tumor location,tumor size,depth of infiltration and TNM staging(P > 0. 05). Conclusions:CIMP on chromosome 3p may occur in early stage of oncogenesis of gastric cancer,and influencing tumor differentiation and lymph node metastasis.

Objective To observe the morphological changes of macular capillary in type 2 diabetic mellitus (DM) patients without clinical features of diabetic retinopathy (DR) by optical coherence tomography angiography (OCTA). Methods This is a prospective clinical case-control study. Forty-three eyes of 22 patients with DM without clinical features of DR (case group) and 40 control eyes of 20 age-and sex-matched healthy physical examination subjects (control group) were enrolled in this study. All subjects underwent OCTA examination with mode of retinal blood flow imaging, macular 3 mm×3 mm and 6 mm×6 mm area, signal strength>45. Foveal avascular zone (FAZ) area, foveal capillary density, parafovea capillary non-perfusion, and micro-aneurysm in shallow capillary vessel layer were evaluated. Results In case group, the mean FAZ area was (0.397±0.141) mm2 and the mean foveal capillary density was (44.6±0.62)%. In control group, the mean FAZ area was (0.253±0.112) mm2 and the mean foveal capillary density was (48.6±0.58)%. FAZ area of eyes in case group was larger than that in control group (t=1.017, P<0.05). There was no difference of foveal capillary density between two groups (t=1.499, P>0.05). The spider web-like FAZ and normal foveolar avascular zone were observed in eyes of control group. The parafovea capillary non-perfusion, abnormal foveolar avascular zone, micro-aneurysm and tortuosity of vessels were observed in eyes of case group. Parafovea capillary non-perfusion (χ2=4.542), micro-aneurysms (χ2=5.183) were seen more often in case group than control group (P<0.05). Conclusion Type 2 DM patients have abnormal retinal vascular microcirculation before DR using OCTA, including larger FAZ area, parafovea capillary non-perfusion, abnormal foveolar avascular zone, micro-aneurysm and tortuosity of vessels.

Clinical trial protocol is the document that illustrates the background of a clinical trial, theoretic basis, objective, design, methods, and organization, as well as statistical calculating, implement, and conditions for completion. Clinical trial protocol is the basic measure for ensuring the validity of scientific results and reducing bias. In order to optimize the design of clinical trial protocol, we generalize main problems in Chinese medicine clinical trials, key points of clinical trial protocol, as well as report standards.
Clinical Trials as Topic ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; Research Design ; standards

Objective To develop a nylon membrane chip for rapid and systematic detection of the diabetes-associated 45 mutant loci in mitochondrial DNA(mtDNA).Methods The mutant-and wild-type probes were designed for detection of 45 mutant loci in mtDNA with Primer Premier 5.0 and NCBI BLAST softwares and the 90 probes with 8 poly T were immobilized on the Hybond N~+ nylon membranes which were treated with 5?SSC Buffer by UV-crosslinking;Then asymmetric PCR was employed to obtain the target single strand DNA(ssDNA).The PCR products were labeled with biotin after purification.NBT/BCIP was used as substrate that yields a very intense purple signal followed by AP-avidin,and the signals were observed in 24 samples with known sequences to evaluate the chips,each sample was repeatedly measured three times.Results The specific target fragments of 45 loci can be amplified under the same condition with nine sets of primers.The annealing temperatures of the wild-type [(59.01?1.42)℃] and mutant-type [(59.34?1.29)℃ ] probes are so close(t=1.046,P =0.301)that hybridization can be performed at the same temperature.The spots on the membrane chip are distinct,regular and well-distributed.The results of positive-and negative-control are perfect.The signals of negative probes and the background are similar.The results of chip were nearly concordant with that of DNA sequences(?~2=113.132,Kappa value =0.888,P = 0.000)except for T16189C mutant.Conclusions We have successfully developed a nylon membrane chip for rapid and systematic detection of the diabetes-associated 44 mutant loci in mtDNA.It could be used for screening for diabetic patients and high-risk people.

Background: Acute graft?versus?host disease (aGVHD) is a common and severe complication of allogeneic hematopoietic stem cell transplantation (allo?HSCT). Some studies have found that the presence of certain specific human leukocyte antigen (HLA) loci could affect the occurrence of aGVHD. Meanwhile, the impact of HLA haplotypes on aGVHD has been rarely studied. This study aimed to investigate the effects of HLA loci and haplotypes on intestinal aGVHD. Methods: Totally, 345 consecutive patients undergoing first HLA?matched sibling peripheral blood stem cell transplantation (PBSCT) from February 2004 to June 2013 at Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, were enrolled in this study. HLA loci and haplotypes of recipients with frequency over 5% were searched and their effects on intestinal aGVHD were investigated. Other important factors including donor age, recipient age, donor?recipient sex combinations, and conditioning regimens were also evaluated using logistic regression. Pure upper gastrointestinal tract aGVHD without diarrhea was excluded because the histological proof was unavailable. The follow?up end?point was 6 months after HSCT. Results:The cumulative incidence of intestinal aGVHD was 19.4%,with 18.0%of the patients classified as classic aGVHD and 1.4% aspersistent, recurrent,or late aGVHD.Multivariate analysis showed that HLA?A31 locus(odds ratio[OR]2.893,95% confidence interval[CI][1.054,7.935], P = 0.039), HLAB40?DR15 (OR 3.133, 95% CI [1.250, 7.857], P = 0.015), and HLAB46?DR9 haplotypes (OR 2.580, 95% CI [1.070, 6.220], P = 0.035), female donor for male recipient (OR 2.434, 95% CI [1.319, 4.493], P = 0.004) were risk factors for intestinal aGVHD. Conclusion: The presence of certain HLA loci and haplotypes may influence the occurrence of intestinal aGVHD in PBSCT with HLA?identical sibling donors.

BACKGROUND:Increasing evidence has shown that insulin-like growth factor-1 (IGF-1) plays a regulatory role in bone metabolism and increases bone formation,stimulates osteoblast number and activity,as well as reduces osteoclast differentiation and bone resorption.The mechanism of IGF-1 is an issue of concern.OBJECTIVE:To review the research advance in IGF-1 and the bone metabolism-related indexes in postmenopausal osteoporosis.METHODS:The first author searched the PubMed and CNKI databases between January 2012 and July 2016 using the keywords of "postmenopausal osteoporosis,insulin-like growth factor,bone metabolism,biochemical markers" in English and Chinese,respectively.The repetitive articles were excluded,and 36 eligible articles were enrolled for overview.RESULTS AND CONCLUSION:Bone resorption is increased in postmenopausal woman through the regulation of a variety of cytokines,in which IGF-1 in the blood is combined with IGF binding protein 3,making growth hormone play its biological role.In addition,the growth hormone in the blood directly acts on the adipose tissue,and muscle and bones.Growth hormone exerts a direct effect on bone tissue,promotes osteoblast maturation and differentiation,and enhances the expression of collagen and non-collagen through IGF-1-mediated indirect effect,thus promoting bone formation.The process of bone metabolism is able to reflect the activities of osteoblasts and osteoclasts and the changes of bone matrix and bone mineralization.In vitro experiments show that IGF-1 stimulates the proliferation of osteoblast precursors and differentiate into osteoblasts in a dose-dependent manner,promotes the expression of osteogenic markers such as alkaline phosphatase,type Ⅰ collagen,and osteocalcin,and also stimulates the activity and number of osteoclasts.However,there are few clinical reports and few observation indicators,resulting in a lack of reference range for the detection and treatment of osteoporosis,which needs further exploration.

12E7 Antigen ; Antigens, CD ; metabolism ; Bone Neoplasms ; metabolism ; pathology ; surgery ; Cell Adhesion Molecules ; metabolism ; Chondrosarcoma, Mesenchymal ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Hemangiopericytoma ; pathology ; Humans ; Lung Neoplasms ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Pneumonectomy ; methods