Objective To use superparamagnetic iron oxide nanoparticles(SPIONs)to label chimeric antigen receptor(CAR)T cells targeting carcinoembryonic antigen(CEA),and perform magnetic resonance imaging(MRI)to real time trace CEA CAR-T cells in vivo.Methods Appropriate amount of ferumoxytol,heparin sodium and protamine sulfate were mixed at high(ferumoxytol 100 μg/mL,heparin sodium 4 IU/mL,protamine sulfate 120 μg/mL),medium(ferumoxytol 50 μg/mL,heparin sodium 2 IU/mL,protamine sulfate 60 μg/mL),and low(ferumoxytol 25 μg/mL,heparin sodium 1 IU/mL,protamine sulfate 30 μg/mL)concentrations to form a SPIONs complex ferumoxytol/heparin/protamine(FHP),and then co-incubated with CEA CAR-T cells for cell labeling.The biocompatibility of FHP was detected by CCK-8 assay,EdU assay and flow cytometry.The uptake of FHP was detected by Prussian blue staining,and SPIONs content in the cells was quantitatively detected by inductively coupled plasma-mass spectrometry(ICP-MS).Flow cytometry was used to detect the lytic effect of FHP-labeled CEA CAR-T cells on tumor cells,and MRI was employed to scan FHP-labeled CEA CAR-T cells.Results FHP at high,medium,and low concentrations had no significant effect on the activity of CEA CAR-T cells,with cell activity above 100％determined by CCK-8 assay.DNA proliferation was above 94.3％in EdU assays.Prussian blue staining showed that CEA CAR-T cells could take FHP up,with the uptake increased with the increment of FHP concentration.ICP-MS showed that the intracellular Fe content was 440.23±189.36 ng/mL.Tumor cell killing experiment showed that FHP-labeled CEA CAR-T cells had excellent killing capability against tumor cells.MRI scans indicated that T2WI signals of FHP-labeled CEA CAR-T cells were significantly reduced with increasing FHP concentration(P＜0.01).Conclusion SPIONs complex FHP shows good biocompatibility and can effectively label CEA CAR-T cells.SPIONs complex FHP can be used as a magnetic marker for CEA CAR-T cells and a feasible MRI tracer for clinical application.

In clinic, intima and media thickness are the main indicators for evaluating the development of atherosclerosis. At present, these indicators are measured by professional doctors manually marking the boundaries of the inner and media on B-mode images, which is complicated, time-consuming and affected by many artificial factors. A grayscale threshold method based on Gaussian Mixture Model (GMM) clustering is therefore proposed to detect the intima and media thickness in carotid arteries from B-mode images in this paper. Firstly, the B-mode images are clustered based on the GMM, and the boundary between the intima and media of the vessel wall is then detected by the gray threshold method, and finally the thickness of the two is measured. Compared with the measurement technique using the gray threshold method directly, the clustering of B-mode images of carotid artery solves the problem of gray boundary blurring of inner and middle membrane, thereby improving the stability and detection accuracy of the gray threshold method. In the clinical trials of 120 healthy carotid arteries, means of 4 manual measurements obtained by two experts are used as reference values. Experimental results show that the normalized root mean square errors (NRMSEs) of the estimated intima and media thickness after GMM clustering were 0.104 7 ± 0.076 2 and 0.097 4 ± 0.068 3, respectively. Compared with the results of the direct gray threshold estimation, means of NRMSEs are reduced by 19.6% and 22.4%, respectively, which indicates that the proposed method has higher measurement accuracy. The standard deviations are reduced by 17.0% and 21.7%, respectively, which indicates that the proposed method has better stability. In summary, this method is helpful for early diagnosis and monitoring of vascular diseases, such as atherosclerosis.
Carotid Arteries/diagnostic imaging* ; Carotid Intima-Media Thickness ; Normal Distribution ; Ultrasonography

OBJECTIVE: To detect potential variant in an ethical Han Chinese pedigree affected with breast cancer. METHODS: The proband and her relatives were subjected to next-generation sequencing using a target capture sequencing kit containing 121 cancer-related genes. Candidate variants were selected by analysis of their type, frequency in population, and segregation with the phenotype. Candidate variant was verified by Sanger sequencing and TA cloning. RESULTS: A c.2013_2014ins GT variant was detected in the BRCA1 gene among all breast cancer patients from this pedigree but not among healthy females. The variant was not recorded in the 1000 Genome Project database or the Exome Aggregation Consortium (ExAC) database. The frameshifting insertion was predicted to form an premature stop codon in gene transcript and can give rise to a truncated protein. CONCLUSION The BRCA1 c.2013_2014ins GT variant probably underlies the pathogenesis of breast cancer in this Chinese pedigree.
Asian Continental Ancestry Group ; BRCA1 Protein ; genetics ; Breast Neoplasms ; genetics ; Exome ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Pedigree ; Phenotype

AIM:To investigate the effect of angiotensin II type 1 receptor (AT1R)-calcineurin (CaN) signa-ling pathway on the expression of sodium current channel Nav 1.5 at mRNA and protein levels in the hypertrophic ventricu-lar myocytes from neonatal rats .METHODS:The ventricular myocytes were isolated from the ventricles of 1-day-old neo-natal Sprague-Dawley rats and were divided into 4 groups according to different drug intervention as control group , pheny-lephrine (PE) group, losartan (Los)+PE group and cyclosporin A (CsA)+PE group.The method of RNA interference mediated by adenovirus carrying short hairpin RNA ( shRNA) was used to knock down the gene which encodes the beta subtype of CaN A subunit (CnAβ) and the cells were divided into 4 groups as Ad-Null group, Ad-Null+PE group, Ad-CnAβshRNA1 group and Ad-CnAβshRNA1+PE group.The mRNA expression of brain natriuretic peptide ( BNP) ,β-my-osin heavy chain (β-MHC) and Nav1.5 was detected by RT-qPCR.The protein levels of CnAβand Nav1.5 in the whole-cell extracts were determined by Western blot analysis .RESULTS:Treatment of the neonatal rat ventricular myocytes with PE for 24 h increased the protein-to-DNA ratio and the mRNA expression of BNP and β-MHC.The size of the cell surface was also increased after PE treatment .Treatment of the cells with PE increased the protein expression of CnAβ, and re-duced the protein expression of Nav 1.5.Both Los and CsA prevented those effects of PE .The mRNA expression of Nav1.5 was reduced by PE , and no significant difference of Nav 1.5 mRNA expression among PE group , Los+PE group and CsA+PE group was observed .Silencing of CnAβin the neonatal rat ventricular myocytes using Ad-CnAβshRNA1 inhibited the ability of PE to increase the mRNA expression of BNP , and diminished the ability of PE to reduce the protein expression of Nav1.5.CONCLUSION:AT1 R-CaN signaling pathway participates in regulating protein expression of Nav 1.5 in the hy-pertrophic ventricular myocytes from neonatal rats .

Objective: To explore the role of angiotensin receptor type I (AT1)-calcineurin (CaN) signaling pathway in transient outward potassium ion channel (Ito) remolding in hypertrophic atrial myocytes of neonatal rats. Methods: 1 day old neonatal rats’ atrial myocytes were isolated and the cells were divided into 4 groups:①Control group, normal cells were cultured for 24 h,②Stretching group, the cells were cultured for 24 h with mechanical stretching to induce hypertrophy,③Telmisartan group, the cells were treated by telmisartan at 1 μmol/L for 1 h, then cultured for 24 h and ④Cyclosporin-A (CsA) group, the cells were treated by CsA at 0.25 μg/ml for 1 h, then cultured for 24 h. The ratios of protein/DNA in myocytes were compared between Control group and Stretching group, cell hypertrophy was deifned by mRNA expression of atrial natriuretic peptide (ANP). Ito changes were detected by whole-cell patch clamping technique, proteins expressions of Kv4.3 and CaN A subunit were examined by Western blot analysis. Results: Compared with Control group, Stretching group showed obviously decreased Ito density and Kv4.3 protein expression, while increased CaN A protein expression; Compared with Stretching group, the above effects were reduced in Telmisartan group and CsA group. Conclusion: AT1-CaN signaling pathway was involved in the regulation of Ito channel remodeling in hypertrophic atrial myocytes of neonatal rats.

OBJECTIVETo explore the relationship between expression of coiled-coil domain-containing protein 80(CCDC80) and obesity in serum and adipose tissues.

METHODSA cross-sectional survey was conducted in a hospital in Tangshan in September 2010. 100 people including 78 healthy people and 22 with type-2 diabetes were recruited. Another 36 female patients with benign tumor of Obstetrics and Gynecology were also recruited. Demographic characteristics and serum samples were collected from all subjects, basic biochemical indicators were tested. All subjects were divided into 'Normal Weight' and 'Overweight and Obese' according to their BMI (BMI <24.0 kg/m(2); BMI≥24 kg/m(2)). Serum CCDC80 of the 100 subjects was detected by ELISA. Visceral and subcutaneous fat were derived from the other 36 female subjects, and Real-time PCR was used to detect CCDC80 mRNA expression in adipose tissues. Pearson correlation and multiple linear regression were used to analyze the correlation between serum or adipose CCDC80 expression and waist, BMI, and other biochemical indicators.

RESULTSThe age of 100 subjects was (52.9±8.4) years old. 44% of them were women (44 cases) and 56% of them were men (56 cases). After dividing them into three groups according to their BMI, covariance analysis were conducted, and age and gender were adjusted. The HDL-C level was significantly different among three groups (F = 10.73, P < 0.001): 'Overweight and obese combined with diabetes' group ((0.90±0.06) mmol/L)< 'Overweight and obese' group ((1.14±0.04) mmol/L) < 'Normal weight' group ((1.28±0.05) mmol/L). The adjusted expression of serum CCDC80 of the 100 subjects was (5.84±0.16) pg/ml, (5.81±0.98) pg/ml among men and (5.97±0.89) pg/ml among women, and there was no significant difference (t = -0.812, P = 0.419) between genders. ANOVA revealed that there was no significant differences of the expression of serum CCDC80 among three groups (F = 1.06, P = 0.351), 'Normal weight' group was (5.84±0.16) pg/ml, 'overweight and obese' group was (6.11±0.14) pg/ml, and 'Overweight and obese combined with diabetes' group was (5.84±0.19) pg/ml. The analysis showed that FBG had a negative correlation with serum CCDC80 (R(b) = -0.223, P = 0.026). Multivariate linear regression had a similar result, with 1 mmol/L increase of serum FBG, serum CCDC80 decreases for 0.24 pg/ml (β = -0.24, 95% CI: -0.21--0.02). There was also a negative correlation between serum CCDC80 and FBG in overweight and obese people (R(a) = -0.368, P = 0.013). Besides, CCDC80 mRNA was detected in both subcutaneous and visceral adipose tissue of 36 cases, the expression level was 0.06±0.02 for subcutaneous fat, was 0.05±0.04 for visceral fat, and the expression in visceral fat was lower (0.05±0.03) than that in subcutaneous fat (0.06±0.03) (t = 2.50, P = 0.025) among overweight and obese group. There was a negative correlation between waist and visceral CCDC80 mRNA expression (r = -0.472, P = 0.035).

CONCLUSIONThis study suggested that CCDC80 may be involved in energy and insulin metabolism, and plays a protective role in obesity and diabetes.

Adipose Tissue ; Cross-Sectional Studies ; Diabetes Mellitus, Type 2 ; Female ; Glycoproteins ; Humans ; Intercellular Signaling Peptides and Proteins ; Intra-Abdominal Fat ; Male ; Middle Aged ; Obesity ; Overweight ; Real-Time Polymerase Chain Reaction ; Serum ; Subcutaneous Fat

Objective To investigate the risk factors for castrate-resistant prostate cancer (CRPC) after prostate cancer treated with androgen deprivation therapy (ADT) within 1 year.Methods One hundred and thirty-one prostate cancer patients treated with ADT in our institute between Jan.2008 and Jan.2011 were selected for this study.Patients were followed up by telephone or in clinic,including serum testosterone,serum PSA,clinical symptoms,imaging studies,digital rectal examination (DRE),survival data,PSA nadir,time to PSA nadir and et al.We mainly studied the CRPC after prostate cancer treated with ADT within 1 year.In the 131 patients,the median age was 70 (ranged from 44-89) years.There were 13 patients (9.9％) less than 60 years,43 patients (32.8％) between 60 and 69 years,62 patients (47.3％) between 70 and 79 years,13 patients (9.9％) more than 80 years.The average body mass index (BMI) was 23.0 (ranged from 14.4-34.4) kg/m2.There were 10 patients less than 18.5 kg/m2,77 patients between 18.5 and 24.0 kg/m2,34 patients between 24.1 and 28.0 kg/m2,and 10 patients more than 28.0 kg/m2.The initial PSA was between 0.3 and 4 707.0 μg/L,there were 19 patients (14.5％) less than 20 μg/L,45 patients (34.4％) between 20 and 99 μg/L,67 patients (51.1％) more than 100 μg/L.One patient (0.7％) was in T1,39 patients (29.8％) in T2,59 patients (45.0％) in T3,32 patients (24.4％) in T4.5 patients (3.8％) were with Gleason score 4,13 patients (9.9％) were with Gleason score 5,24 patients (18.3％) were with Gleason score 6,51 patients (38.9％) were with Gleason score 7,26 patients (19.8％) were with Gleason score 8,9 patients (6.9％) were with Gleason score 9,3 patients (2.3％) were with Gleason score 10.Results There were 32 of 131 patients (24.4％) progressed to CRPC after treated with ADT within 1 year.In the CRPC group,there were 3 patients less than 60 years,15 patients between 60 and 69 years,12 patients between 70 and 79 years,2 patients more than 79 years; 3 patients were less than 18.5 kg/m2,19 patients were between 18.5 and 24.0 kg/m2,7 patients were between 24.0 and 28.0 kg/m2,3 patients were more than 28.0 kg/m2 ; 4 patients were less than 20 μg/L,6 patients were between 20 and 100 μg/L,22 patients were more than 100 μg/L; 4 patients were in T2,13 patients were in T3,15 patients were in T4; 2 patients were with Gleason score 6,11 patients were with Gleason score 7,11 patients were with Gleason score 8,6 patients were with Gleason score 9,2 patients were with Gleason score 10; 29 patients were with metastasis,3 patients without metastasis.Clinical stage (P =0.001),Gleason score (P＜0.001) and metastasis (P=0.011) were statistically significant between the CRPC within 1 year group and the rest group.Conclusions The clinical stage and Gleason score are the risk factors of CRPC treated with ADT within 1 year.The higher of the clinical stage and Gleason score,the greater risk to be the CRPC within 1 year.

Objective To apply the nursing intervention based on the protection motivation theory in community elderly patients with hypertension , in order to establish a health behavior and to control bloodpressure and enhance better quality of life .Methods Using the random number table method , 144 elderly patients with hypertension were randomly divided into control group and intervention group .The intervention group received comprehensive nursing intervention based on protection motivation theory .The physiological index, health behavior and the scores of protection motivation questionnaire for elderly patients with hypertension were compared between the control group and the intervention group .Results Before the intervention , the physiological index , health behavior and the scores of protection motivation questionnaire for elderly patients with hypertension between groups had no significant difference (P>0.05).For the intervention group after 6-month-intervention, the systolic blood pressure was( 136.85 ±15.32 ) mmHg, the diastolic blood pressure was (80.78 ±10.54) mmHg, the cholesterol level was (4.49 ±0.83) mmol/L, the score of self-efficacy was (18.03 ±1.75) and the score of susceptibility was (18.32 ±3.04).Those values and scores in the control group were (139.28 ±13.44)mmHg, (89.25 ±8.78) mmHg, (5.33 ±0.91) mmol/L, (7.24 ±1.04), (16.82 ±1.97) and (16.56 ±4.22).The differences between groups were statistically significant (t=1.35, 5.17, 4.98, 3.36, 3.69, 2.11, respectively;P <0.05 ).After 6 months of comprehensive nursing intervention , the physiological index , health behavior and the scores of protection motivation questionnaire for elderly patients with hypertension were statistically significant ( P <0.05).Conclusions Comprehensive nursing intervention based on protection motivation theory can promote the health behavior of community elderly hypertensive patients and can effectively control high blood pressure and improve patients ’ quality of life.

To investigate the influence of Drynaria total flavonoids on proliferation and apoptosis of osteoblasts in tumor necrosis factor-α (TNF-α)- mediated medium, so as to explore the mechanism of Drynaria total flavonoids in preventing and treating osteoporosis of rheumatoid arthritis.

Objective To investigate high risk human papillomavirus(HR-HPV)prevalence among married women in Beijing and to study the high risk flactors.nethods During March 2007 to September 2008.a total of 6185 married women sampled from 137 communities in 12 districts were screened bv HR-HPV DNA test and cytogical test.The interview was carried out with unified questionnaires.The databage was set up and twice entered in EpiDam 3.0.After checked up,the data were analyzed in SPSS 15.0.Results (1) The HR-HPV infection rate was 9.89%.The HR-HPV infection rate of the city zone,the suburb and the exurb were 9.34%,10.51% and 9.51% (P>0.05).The HR-HPV infection rate of the native and the oudander were 9.53%,11.30% (P<0.05).(2) The age distribution of HR-HPV infection was that the rate was around 10% among 25 to 44 age groups,which was the highest(11.21%) in 30 to 34 age group;then the rate was descended as the age raising,the rate of 50 to 54 age group was the lowest(7.78%).(3) Multiple logistic regression showed that the related risk factors of HR-HPV infection mainly included 1000 RMB and above of family income per person per month.possessing more than 1 sexual partner of her husband,outlander and hish levels of education.(4) The prevalence of cervical intraepithelial neoplasia(CIN)in HR-HPV positive group wag significantly higher than that in HR-HPV negative group(29.76% vs 3.32%,P<0.01).Conclusions(1)The HR-HPV infection rate among aged 25 to 54 years was 9.9% and there was no significant difference in area distribution.(2)The hish risk population which should strengthen screening was the married bearing-age women with high level of family income,outlander,high levels of education and her husband possessing more than 1 sexual partner.(3)HR-HPV infection is the main risk factor for CIN and cervical cancer.while does not provide a causal relationship with them.The high risk population should be checked regularly to understand the development of HR-HPV infection and CIN incidence.