Objective To explore the association between the sortilin-related receptor,L (DLR class) A repeats containing (SORL1) gene single nucleotide polymorphisms (SNPs) and sporadic Alzheimer's disease (SAD) in Han Chinese populations.Methods Five candicate SNPs,including rs985421,rs12364988,rs4598682,rs3781834,and rs3781836,within the SORL1 gene were genotyped by LDR-PCR methods,and the association of these SNPs with SAD risk were also analyzed in a case-control study with 179 SAD and 106 healthy controls.Results The results indicated that the A allele frequency of the SORL1 SNP rs985421 was significant difference between SAD and healthy controls (Trend test:P =0.0044,P adj =0.022; Allelic:P=0.0023,P adj =0.012).In subjects without APOE s4 allele,higher frequency of the A allele of rs985421 was observed in SAD patients compared with control subjects by the stratified analysis (OR=2.260,95％ CI =1.269-4.024,P=0.0048).In addition,logistic regression analysis further revealed that the A allele of SORL1 SNP rs985421 was significantly associated with SAD risk (OR=1.968,95％ CI =1.273-3.042,P=0.002).However,no positive signals in other four SNPs within the SORL1 gene were observed (all P＞ 0.05).Conclusions The A allele of SORL1 SNP rs985421 may be an APOE ε4-independent factor associated with SAD risk.

Objective To evaluate the feasibility of Mycobacteria growth indicator tube ( MGIT ) liquid culture combined with rifampin resistance test real-time PCR ( Xpert MTB/RIF test) in the diagnosis of tuberculosis.Methods 652 cases of suspected pulmonary tuberculosis from October 2014 to March 2015 in Quzhou People′s Hospital were enrolled. The morning sputum samples were collected for acid-fast staining, L?wenstein-Jensen ( L-J) culture, BACTEC MGIT culture and Xpert MTB/RIF test.Samples with positive results of MGIT liquid culture were subjected to strain identification and drug sensitivity test with fluid method.Methodological comparison between four methods was made and the performance of MGIT liquid culture combined with Xpert MTB/RIF test in the rapid diagnosis and drug resistance detection in Mycobacterium tuberculosis was evaluated by chi-square test. Results Among the samples from 399 confirmed tuberculosis patients, the diagnostic sensitivity of the 4 methods ( acid-fast staining, L-J culture, MGIT culture and Xpert MTB/RIF test) were 17.0%(68/399), 23.8%(95/399), 37.8% (151/399) and 37.3%(149/399) respectively.The sensitivity and specificity of MGIT culture combined with Xpert MTB/RIF test was 39.8%(159/399) and 94.8%(240/253).The sensitivity of MGIT culture combined with Xpert MTB/RIF test was significantly higher than acid-fast staining (χ2 =50.9, P<0.01 ) and L-J culture (χ2 =23.7,P<0.01).The average detection time of MGIT culture was 7.5 days ( smear positive and Xpert MTB/RIF positive), 13.4 days (smear negative and Xpert MTB/RIF positive) and 16.9 days ( smear negative and Xpert MTB/RIF negative) .The sensitivity and specificity of Xpert MTB/RIF test in rifampin resistance detection were 9/9 and 97.3% ( 129/132 ) respectively.The average detection time of fluid method for drug sensitivity test was 8.3 days.Conclusions MGIT liquid culture combined with Xpert MTB/RIF test can detect Mycobacterium tuberculosis and the drug sensitivity rapidly.The method is highly sensitive and specific.It is important for clinical diagnosis and treatment of tuberculosis.

Objective To evaluate the nutritional status of iodine of pregnant and lactating women in coastal and inland areas of Zhejiang province,and to provide scientific basis for iodine supplementation.Methods In 2009,five coastal cities,five coastal counties and three inland areas from Zhejiang province were selected by the cluster sampling method,and one sub-district(township) was randomly selected in each city (county),then one community (village) was randomly selected in each sub-district (township).Thirty pregnant women,thirty lactating women and fifty children aged 8 - 10,were randomly selected in each community(village) as the investigation objects to collect instant urine samples to detect urine iodine,and to collect thirty resident edible salt and some water samples to detect salt iodine and water iodine,if the numbers of water source were more than 5 then 1 water sample was collected by their location of east,south,west,north and center in each community(village),otherwise,the numbers of water samples collected were equal to the number of community (village).Results The medians of salt iodine of coastal city,coastal county and inland area were 28.42,25.29,and 33.24 mg/kg,respectively,and the medians of water iodine correspondingly were 5.41,11.52,and 2.80 μg/L,respectively.The medians of urinary iodine of pregnant and lactating women in Zhejiang province were 148.72 and 161.34 μg/L,respectively,which were less than that of children aged 8 - 10(231.02 μg/L,Z =8.57,8.24,all P ＜ 0.017).The medians of urinary iodine in pregnant women from different areas were ranked in order as inland areas(211.42 μg/L),coastal counties (150.51 μg/L) and coastal cities (123.33 μg/L,Z =2.80,5.31,2.42,all P ＜ 0.017),and in lactating women,the medians of urinary iodine in inland areas( 197.43 μg/L) was higher than that in coastal cities( 139.64 μg/L,Z =4.03,P ＜ 0.017).Conclusions The overall levels of iodine nutrition in lactating and pregnant women in Zhejiang province are in the appropriate level,but that of the pregnant women in the coastal cities is inadequate.

OBJECTIVETo investigate the effects of Mycobacterium tuberculosis on apoptosis of mouse dendritic cells (DC 2. 4) and the activation of caspase-3, caspase-8.

METHODSMycobacterium tuberculosis H37Rv strain was co-cultured with DC 2. 4 cells. The morphological changes of DC 2. 4 cells were observed with fluorescence microscope after DAPI staining and transmission electron microscope. The apoptosis of DC 2. 4 cells were examined by DNA agarose gel electrophoresis. The activities of caspase-3 and caspase-8 were detected by colorimetric assay.

RESULTSBacterial invasion was observed while DC 2. 4 cells and H37Rv were co-cultured for 2 h; and the rates of invasion were (16.1 ± 4.3)%, (35.8 ± 5.1)%, (50.2 ± 5.7)%, (58.3 ± 6.2)% and(65.9 ± 6.9)% at 4, 6, 8,10, 12 h, respectively. The phenomenon of nuclear condensation and marginalization were shown by DAPI staining and transmission electron microscope in DC 2. 4 cells at 6 h of co-cultivation with H37Rv. The characteristic bands of apoptosis by DNA electrophoresis were detected. The activities of caspase-3 and caspase-8 were increased in a time-dependent manner. The rates of DC 2. 4 cell apoptosis were (6.4 ± 2.5)%, (11.8 ± 5.3)% and (31.1 ± 8.7)% at 6 h,12 h and 24 h after co-cultivation with H37Rv, respectively. The maximal activities of intracellular caspase-3 and caspase-8 at 10 h and 6 h were (2.01 ± 0.09) U/μg and (2.40 ± 0.07)U/μg, respectively, which was significantly different compared with the control groups(P<0.05). The activation of caspase-8 was earlier than that of caspase-3.

CONCLUSIONMycobacterium tuberculosis can induce the apoptosis of DC 2. 4 cells, which is associated with the activation of intracellular caspase-3 and caspase-8.

Animals ; Apoptosis ; physiology ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Cells, Cultured ; Dendritic Cells ; metabolism ; pathology ; Mice ; Mycobacterium tuberculosis

Objective To evaluate the dietary iodine intake level of population in coastal and inland areas of Zhejiang province.Methods The cluster sampling method was applied to select Zhoushan,Ningbo and Taizhou cities from Zhejiang province as coastal areas,to select Jinhua,Quzhou and Lishui cities from Zhejiang province as inland areas,and two counties(districts) were randomly selected in each chosen city,three sub-districts(towns) were randomly selected in each chosen counties (districts),then one community (village) was randomly selected in each chosen sub-districts(towns).Adult residents aged greater than 18 were selected as investigation subjects,their dietary iodine intake in the past 24 hours was investigated,and the Food composition table” was inquired to get the data of dietary iodine intake.One hundred copies of residential edible salt samples and drinking water samples in each of the selected community (village) were collected to detect salt iodine and water iodine by direct titrimetric and spectrophotometric method,respectively.Results The mean of adult residents' dietary iodine intake in Zhejiang coastal area was 267.76 μg/d,which was less than that in inland area (429.05 μg/d,t =- 6.90,P ＜0.05),and the means of adult residents' dietary iodine intake from drinking water,laver and fish(5.75,69.72,5.61 μg/d,respectively) in coastal area were higher than those in inland areas(3.25,35.27,3.43 μg/d,respectively,t =21.73,3.92,4.08,all P ＜ 0.05),however,the means of adult residents' dietary iodine intake from salt,kelp and other food (166.81,3.04,16.82 μg/d,respectively) in coastal areas were less than those in inland areas (355.15,6.14,25.81 μg/d,respectively,t =- 8.76,- 5.49,- 18.56,all P ＜ 0.05).In coastal areas,the proportion which was less than estimated average requirement of iodine(EAR,120 μg/d) was 46.48％ (1029/2214),the ratio which was higher than the maximum tolerable intake of iodine(UL,1000 μg/d) was 3.34％ (74/2214),and the average contribution rate of dietary intake of iodine in salt was 62.30％(166.81/267.76).While in inland areas,the corresponding proportions were 7.61％ (171/2246),2.80％ (63/2246) and 82.78％ (355.15/429.05),respectively.Conclusions The dietary iodine intake in Zhejiang inland areas has reached the recommended nutrient intake levels of the Chinese Nutrition Society,but there is a certain degree of insufficient iodine intake in population of the coastal areas.

OBJECTIVE: To investigate the relationship between the clinical features of carotid transient ischemic attacks (TIA) and the intracranial or extracranial angiostenosis. METHODS: Location and degree of stenosis of involved arteries were examined by the digital subtraction angiography in 52 patients with carotid TIA. RESULTS: Intracranial or extracranial vascular lesions of different degrees were revealed in 45 patients (86.5%), and 29 out of 45 (64.4%) had more than one site. Severe stenosis and occlusion occurred more frequently in TIA patients with short duration (less than 1 hour) and multiple attacks (more than twice). CONCLUSION Most patients with TIA of carotid systems have stenosis in intracranial or extracranial arteries. TIA with short duration and multiple attacks always accompany with severe stenosis or occlusion in intracranial or extracranial arteries. Digital subtraction angiography helps to identify the vascular etiology of TIA and provides the instruction of therapeutic regimen.
Adult ; Aged ; Angiography, Digital Subtraction ; Carotid Stenosis ; complications ; diagnostic imaging ; pathology ; Cerebral Angiography ; Cerebrovascular Disorders ; complications ; diagnostic imaging ; pathology ; Female ; Humans ; Ischemic Attack, Transient ; diagnostic imaging ; pathology ; Male ; Middle Aged

BACKGROUNDCarotid artery stenting (CAS) as a competing treatment modality has had to adhere to limits to gain widespread acceptance in some studies. This study analyzed the clinical data of 1700 consecutive patients after CAS to retrospectively evaluate the 30-day outcome of CAS for internal carotid artery stenosis in a Chinese population.

METHODSMedical records of 1700 patients who underwent CAS at Xuanwu Hospital affiliated to Capital Medical University between January 2001 and August 2012 were reviewed. Postoperative 30-day complication rates were analyzed and compared with those of other studies. Univariate and multivariate Logistic regression analyses were used to identify factors associated with perioperation myocardial infarction (MI), stroke, and death.

RESULTSThe overall 30-day rate of MI, stroke, and death after CAS was 2.53%. In univariate analysis, patients who were symptomatic, had a neurological deficit (modified Rankin score (mRS) ≥3; P = 0.001), and who were not taking statins experienced a significantly increased rate of MI, stroke, and death (P = 0.017). In multivariate Logistic regression analysis, the presence of symptoms (odds ratio (OR) = 2.485; 95% confidence interval (CI) = 1.267-4.876; P = 0.008) and a neurological deficit (mRS ≥3) (OR = 3.025; 95% CI = 1.353-6.763; P = 0.007) were independent risk factors for perioperative MI, stroke, and death.

CONCLUSIONSAccording to this single-center experience, CAS may effectively prevent and treat carotid artery stenosis that would otherwise lead to stroke. Being symptomatic and having a neurological deficit (mRS ≥3) increased the risk of perioperative MI, stroke, and death.

Adult ; Aged ; Aged, 80 and over ; Carotid Stenosis ; surgery ; Female ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Myocardial Infarction ; Stents ; Stroke ; surgery ; Treatment Outcome

OBJECTIVETo evaluate hemodynamic character and surgical effect of stenoses or occlusion of carotid artery system by perfusion-weighted MRI.

METHODSTwenty-one patients with stenoses or occlusion of carotid artery system underwent surgical treatment. Two patients underwent endarterectomy, extracranial-intracranial arterial bypass to 10 patients, and percutaneous stent angioplasty to 9 patients.

RESULTSPreoperative perfusion-weighted magnetic resonance (MR) revealed normal regional cerebral blood flow in all patients, with delayed time to peak in both middle cerebral artery distribution and border zone. Postoperative perfusion-weighted MR revealed normal time to peak in border zone, but abnormal in middle cerebral artery distribution. Evaluated by perfusion-weighted MR, it showed that surgical method can improve the hemodynamic disorder of this kind of disease. The early curative effect of endarterectomy and percutaneous stent angioplasty is better than extracranial-intracranial arterial bypass.

CONCLUSIONSPerfusion-weighted MR is a good method to evaluate hemodynamic character of stenoses or occlusion of carotid artery system. Surgical method is helpful to this kind of disease, and its curative effect can be evaluated impersonally and accurately by this new technique.

Adult ; Aged ; Angioplasty ; Carotid Stenosis ; physiopathology ; surgery ; Cerebrovascular Circulation ; Endarterectomy, Carotid ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Angiography ; methods ; Male ; Middle Aged ; Sensitivity and Specificity ; Treatment Outcome

OBJECTIVETo investigate the inhibitory effects of an antisense PC cell derived growth factor (PCDGF) vector on proliferation and invasion of highly malignant ovarian cancer cell lines Sw626 and A2780 cells, and preliminarily explore the related mechanisms.

METHODSMTT assay and Boyden chamber in vitro invasion assay were employed to detect the changes of proliferation and invasion ability in the Sw626 and A2780 cells transfected with anti-sense PCDGF. The expression levels of cyclin D1 and CDK4 proteins before and after transfection were detected by Western blotting. The effects on the expression and activity of MMP-2 were evaluated by quantitative RT-PCR and zymography, respectively.

RESULTSComparing with the blank group, the proliferation inhibition rate of the Sw626 and A2780 cells transfected with anti-sense PCDGF was 72.9% and 70.9%, respectively, and the invasion ability was inhibited by 62.9% and 59.0%, respectively. The levels of cyclin D1 and CDK4 protein expression in antisense PCDGF transfected cells were 0.38 +/- 0.08 and 0.37 +/- 0.13, respectively, all significantly lower than 0.84 +/- 0.11 and 0.64 +/- 0.11, respectively, in the blank group (P < 0.01). The MMP-2 mRNA expression level in antisense PCDGF transfected cell group was 0.66 +/- 0.11, not significantly decreased in comparison with 0.89 +/- 0.09 in the blank group (P > 0.05), but the activity of MMP-2 was inhibited significantly.

CONCLUSIONThe antisense PCDGF vector may inhibit markedly the proliferation and invasion of highly malignant ovarian cancer cells, and partially reverses their malignant phenotype. It seems to be related with down-regulating the expression of cyclin D1 and CDK4 and inhibiting the activity of MMP-2. Our findings indicate that PCDGF may become a new target for antisense gene therapy of ovarian cancer.

Cell Adhesion ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; DNA, Antisense ; Down-Regulation ; Female ; Genetic Vectors ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Neoplasm Invasiveness ; Ovarian Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Transfection

BACKGROUNDA satisfactory animal model of breast cancer metastasizing to bone is unavailable. In this study, we used human breast cancer stem-like cells and human bone to build a novel "human-source" model of human breast cancer skeletal metastasis.

METHODSHuman breast cancer stem-like cells, the CD44+/CD24-/lower subpopulation, was separated and cultured. Before injection with the stem-like cells, mice were implanted with human bone in the right or left dorsal flanks. Animals in Groups A, B, and C were injected with 1 x 10(5), 1 x 10(6) human breast cancer stem-like cells, and 1 x 10(6) parental MDA-MB-231 cells, respectively. A positive control group (D) without implantation of human bone was also injected with 1 x 10(6) MDA-MB-231 cells. Immunohistochemistry was performed for determination of CD34, CD105, smooth muscle antibody, CD44, CD24, cytokine, CXC chemokine receptor-4 (CXCR4), and osteopontin (OPN). mRNA levels of CD44, CD24, CXCR4, and OPN in bone metastasis tissues were analyzed by real-time quantitative polymerase chain reaction (PCR).

RESULTSOur results demonstrated that cells in implanted human bones of group B, which received 1 x 10(6) cancer stem-like cells, stained strongly positive for CD44, CXCR4, and OPN, whereas those of other groups showed no or minimum staining. Moreover, group B had the highest incidence of human bone metastasis (77.8%, P = 0.0230) and no accompaniment of other tissue metastasis. The real-time PCR showed an increase of CD44, CXCR4, and OPN mRNA in metastatic bone tissues in group B compared with those of groups C and D, however the expression of CD24 mRNA in group B were the lowest.

CONCLUSIONSIn the novel "human source" model of breast cancer, breast cancer stem-like cells demonstrated a higher human bone-seeking ability. Its mechanism might be related to the higher expressions of CD44, CXCR4, and OPN, and the lower expression of CD24 in breast cancer stem-like cells.

Animals ; Bone Neoplasms ; secondary ; Breast Neoplasms ; pathology ; CD24 Antigen ; analysis ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Hepatocyte Growth Factor ; pharmacology ; Humans ; Hyaluronan Receptors ; analysis ; Immunohistochemistry ; Mice ; Neoplastic Stem Cells ; pathology ; Osteopontin ; analysis ; Phenotype ; Receptors, CXCR4 ; analysis