Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*

The study of atrial fibrillation (AF) has been known as a hot topic of clinical concern. Body surface potential mapping (BSPM), a noninvasive electrical mapping technology, has been widely used in the study of AF. This study adopted 10 AF patients' preoperative and postoperative BSPM data (each patient's data contained 128 channels), and applied the autocorrelation function method to obtain the activation interval of the BSPM signals. The activation interval results were compared with that of manual counting method and the applicability of the autocorrelation function method was verified. Furthermore, we compared the autocorrelation function method with the commonly used fast Fourier transform (FFT) method. It was found that the autocorrelation function method was more accurate. Finally, to find a simple rule to predict the recurrence of atrial fibrillation, the autocorrelation function method was used to analyze the preoperative BSPM signals of 10 patients with persistent AF. Consequently, we found that if the patient's proportion of channels with dominant frequency larger than 2.5 Hz in the anterior left region is greater than the other three regions (the anterior right region, the posterior left region, and the posterior right region), he or she might have a higher possibility of AF recurrence. This study verified the rationality of the autocorrelation function method for rhythm analysis and concluded a simple rule of AF recurrence prediction based on this method.

BACKGROUND: Both aldehyde dehydrogenase 2 (ALDH2) rs671 polymorphism and lifestyle behaviors are involved in coronary artery disease (CAD), while the interaction between them is currently unknown. METHODS: A nested case-control study was conducted in 161 patients with CAD and 495 controls in dyslipidemia population in Yinzhou District, Ningbo, Zhejiang Province, China, in August 2013. Anthropometric data and blood samples were collected, demographic characteristics and lifestyle behaviors information were obtained by a face-to-face interview, dietary intake was assessed by a food frequency questionnaire, and genomic DNA was genotyped. RESULTS: Carriers with increasing number of A alleles had an elevated CAD risk compared with G allele carriers (adjusted OR = 1.483, 95% CI = 1.114-1.974). Carriers of rs671 A/G and A/A genotypes had a higher CAD risk than carriers of G/G genotype (adjusted OR = 1.492, 95% CI = 1.036-2.148). Similarly, individuals with rs671 A/A genotype had a higher CAD risk than individuals with A/G and G/G genotypes (adjusted OR = 2.161, 95% CI = 1.139-4.101). We found a borderline additive interaction between regular fried food intake and A/A and A/G genotypes, and a significantly additive interaction between sedentary/light physical activity and A/A and A/G genotypes. CONCLUSIONS Individuals with A/A or A/G genotypes of rs671 have a higher CAD risk, if they lack physical activity and take fried food regularly, than individuals with G/G genotypes. These findings can help to provide a guide to targeted heart health management.
Adult ; Aged ; Aged, 80 and over ; Aldehyde Dehydrogenase, Mitochondrial ; genetics ; Alleles ; Case-Control Studies ; China ; Coronary Artery Disease ; blood ; genetics ; Dyslipidemias ; blood ; genetics ; Female ; Genetic Association Studies ; Genetic Predisposition to Disease ; Genotype ; Humans ; Life Style ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Risk Factors

Objective: To develop the Chinese primipara social capital scale (C-PSCS), and to evaluate its validity and reliability. Methods: The items of C-PSCS were developed based on Social Capital Scale by the World Bank and Social Network Scale. This scale was modified according to the characteristics of primiparas. We selected 10 experts who specialized in related field, and two rounds of seminars about content, cultural compatibility, primiparas' characteristics, and practicability. The finally C-PSCS included four dimensions: social trust, social reciprocity, social network, and social participation. Using purposive sampling to select 1 100 primiparas in their third trimesters (gestational weeks from 30 to 36 weeks). The validity analyses included content validity, construct validity and discriminant validity. The reliability analyses included Cronbach' α coefficient, and split-half reliability. Results: 1 035 questionnaires (94.09%) were qualified and the completion time was (13.23 ± 2.53) minutes. The total score of scale was 195.38 ± 45.98, and scores for social trust, social reciprocity, social network, and social participation were 30.26 ± 4.25, 22.84 ± 4.21, 34.23 ± 7.47, and 108.05 ± 41.96, respectively. The common factor cumulative variance contribution rates of each dimension were from 52.92% to 69.37%, which achieved more than 50% of the approved standard, and all the items held factor loading >0.5 in its relevant common factor, it had good construct validity. The scale had a good content validity, the r values between each dimensions and total scale were range from 0.57 to 0.81, P<0.01. For the high-score group, scores for social trust, social reciprocity, social network, and social participation dimensions were 32.89±3.19, 25.65±3.48, 38.27±6.59, 119.94±36.61, respectively, which were higher than low-score group (27.77±3.58, 20.18±2.91, 30.40±6.13, 96.76 ± 43.60), t-values were -24.23, -27.46, -19.90, and -9.24, respectively, P<0.001. It had good discriminant validity. The Cronbach'α coefficient for the total scale and four dimensions were from 0.76 to 0.86, and whose split-half reliability were from 0.68 to 0.84. Conclusion The validity and reliability of C-PSCS were excellent, and could be used to evaluate the social capital situation on the Chinese primiparas.

Objective: To investigate the situation of social support (SS), and explore its relationship with antenatal depression (AD) among Zhejiang primiparas in their third trimesters. Methods: From March to August 2016, a cross-sectional study was conducted and the questionnaire was used at the outpatient consulting room of one maternity hospital in Hangzhou. Inclusion criteria includes the primiparas over 18 years old, gestation from 30 to 36 weeks, been able to understand and complete the questionnaires independently, no family history and history of mental disorders and no use of psychotropic drugs, without serious illness and so on. Exclusion criteria was that the primiparas were unable to complete all the contents of the questionnaire and of poor compliance. 1 150 questionnaires were actually given out, and 1 075 questionnaires were valid, so the valid rate was 93.48%. AD was evaluated by the Edinburgh Postnatal Depression Scale (EPDS) and SS was evaluated by the Perceived Social Support Scale (PSSS). SS between the non-AD group and the AD group was compared. The correlation between SS and AD was analyzed. Binary logistic regression model was used to assess the relationship between SS and AD. The level of SS was divided by average scores, groups lower than the average score was defined as the low-score group, groups higher than the average score was defined as the high-score group. Results: The prevalence of AD (score≥9) was 27.3% (293/1 075) among Zhejiang primiparas in their third trimesters. The scores of family support, friend support and other support, and the total score of SS among the non-AD group were 24.80±2.83, 23.40±3.00, 21.91±3.54 and 70.11±7.92, respectively, which were higher than those in the AD group (22.71±3.88, 21.45±3.59, 19.95±3.97, 64.10±10.01), (t values were 8.43, 8.29, 7.83 and 9.25, respectively, P<0.001 for all). The scores of family support, friend support and other support, and the total score of SS were negatively correlated with AD (rs values were-0.26,-0.25,-0.22 and-0.28, respectively, P<0.001 for all). Compared with low-score group, the scores of family support, friend support and other support, and the total score of SS among the high-score group had a lower risk of antenata depression among primiparas in their third trimesters, OR(95%CI) values were 0.56 (0.41-0.77), 0.66(0.47-0.92), 0.57(0.41-0.79) and 0.36(0.27-0.48), respectively. Conclusion The prevalence of AD among Zhejiang primiparas was relatively high, and AD was negatively associated with SS level. We suggest adding SS in community pregnancy health management service in the future.

OBJECTIVETo establish a two-step pretargeting approach to lymphoma radioimmunoimaging in mice using biotinynaled CD45 monoclonal antibody (McAb) and (188)Re-avidin in a tumor-bearing mouse model.

METHODSSix Nod-Scid mice bearing lymphoma cell xenograft were randomized to receive either an intravenous injection of 50 µg/200 µL biotinyled CD45 McAb followed 24 h later by an intraperitoneal injection of 3.7 MBq (50 µg/100 µL) (188)Re-avidin (two-step pretargeting group), or a single intravenous injection of 3.7 MBq (100 µg/100 µL) (188)Re-CD45 McAb (control group). SPECT was performed at 0.5, 1, 6 and 23 h post-injection to characterize (188)Re isotope biodistribution. At 24 h pos-injection, the mice were sacrificed for measurement of radioactivity uptake in the tumor and normal tissues and calculation of the tumor-to-non-tumor (T/NT) ratios.

RESULTSSPECT showed that the two-step pretargeting method resulted in a low radioactivity in the blood pool during the imaging and a concentrated radioactivity in the liver and spleen. The transplanted tumor began to be displayed at 1 h post-injection and was clearly displayed at 1-6 h; the images were clear even at 23 h. With the two-step pretargeting method, the radioactive uptake at 24 h post-injection were (1.34∓0.52)%, (6.77∓2.32)%, and (2.81∓1.25)% in the tumor, kidney and liver, respectively, with low radioactivity levels in other organs and high tumor/blood and tumor/muscle ratios (4.28∓0.82 and 8.00∓0.88, respectively). In the control group, SPECT revealed intense radioactivity in the liver, spleen, and kidneys with obscure display of the tumor; at 20 h, the radioactivity in the blood pool remained high but that in the tumor was low, and the tumor/blood and tumor/muscle ratios at 24 h were only 0.58∓0.06 and 3.21∓0.24, respectively.

CONCLUSIONCompared with (188)Re-CD45 McAb, the two-step pretargeting approach exhibits a good specificity in targeting lymphoma with an increased T/NT ratio in mice and allows early tumor display at 1 h post-injection.

Animals ; Antibodies, Monoclonal ; Avidin ; Disease Models, Animal ; Lymphoma ; diagnosis ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Radioimmunodetection ; Tissue Distribution ; Tomography, Emission-Computed, Single-Photon

Objective To study the effect of two-step pretargeting radioimmunotherapy of CD45 McAb and 188Re-Avidin on lymphoma Raji cell line.Methods The CD45 McAb and Avidin were directly labeled with 188Re,and the labeling efficiency and radiochemical purity were measured by the paper chromatography.The specific binding test and competition binding test between 188 Re-CD45 McAb and Raji cells in vitro were also performed.CCK-8 assay was used to determine the inhibition effect on Raji cell proliferation in the pretargeted group,188Re-CD45 McAb,188Re-Avidin and 188ReO4 groups,then the cell survival and proliferation inhibition rate were calculated.Results The specific cell binding rate of 188Re-CD45 McAb with Raji cells was (70.92 ± 1.91) ％,in the competition group,the binging rate of 188Re-CD45 McAb with Raji cells was only (7.96 ± 0.87)％.The Raji cells proliferation was inhibited in all groups with 188Re radiolabel,and the inhibition rate was positively correlated with the radioactivity dose (r=0.907-0.992,P ＜0.05).However,at the same dose,the inhibition effect in the group of two-step pretargeting at each time point were all stronger than those of 188Re-CD45 McAb,188Re-Avidin and 188 ReO4-alone (t =124.76-607.98,P ＜ 0.05).But there was no significantly statistical difference in the inhibitory effect between the groups of 188Re-Avidin and 188ReO4-(P ＞ 0.05).Conclusions It is confirmed that 188Re-CD45 McAb could be specifically bound to Raji cells,and the two-step pretargeting of CD45 McAb and 188Re-Avidin has obvious inhibitory effect on the Raji cell proliferation.

Background and objective Hypoxia is an important biological characteristics of solid tumor, it is not sensitive to radiotherapy and chemotherapy for which is the presence of hypoxic cell, thus increasing their resistance to con-ventional radiotherapy and chemotherapy, therefore, the detection of hypoxia degree of tumor tissue is of great signiifcance. hTe hypoxia imaging of nuclear medicine can relfect the degree of tissue hypoxia, which can selectively retained on the hypoxic cells or tissues, including nitroimidazole and non nitroimidazole; the nitroimidazole is widely and deeply researched as hypoxic celles developer in China and abroad at present. hTe research about application of radionuclide labelled technique has clini-cal application value to develop the hypoxia imaging agent EDTA-MN complexes which was labeled. To study the feasibility of99mTc by direct labeling method, the radiochemical properties evaluation of99mTc-EDTA-MN, and observe the distribution characteristics of99mTc radiolabeled EDTA-MN in the xenogratf lung cancer nude mice bearing non-small cell lung cancer cell (A549), and provide experimental evidence for its further research and application.MethodshTe radiolabeling of EDTA-MN with99mTc was performed with direct labeling method, respectively, on the reaction dosage (10 mg, 5 mg, 2 mg), stannous chlo-ride dosage (8 mg/mL, 4 mg/mL, 2 mg/mL), mark system pH (2, 4, 5, 6) one by one test, using orthogonal design analysis, to ifnd the optimal labeling conditions. Labelling rate, radiochemical purity, lipid-water partition coeffcient and in vitro stability in normal saline (NS) were determined by TLC and HPLC, and the preliminary study on the distribution of99mTc-EDTA-MN in nude mice.Results hTe labeling rate of99mTc-EDTA-MN with the best labeling conditions was (84.11±2.83)%, and the ra-diochemical purity was higher than 90% by HPLC puriifcation, without any notable decomposition at room temperature over a period of 12 h. hTe partition coeffcient was lgP=-3.05, indicated that this complex was hydrophilic. At 3 h post-injection, the imaging of99mTc-EDTA-MN in nude mice bearing non-small cell lung cancer cell showed that more radioactive gathered in bladder at0.5 h, the transplanted tumor was clearly imaged at 1 h post-injection, during whole imaging radioactive in other tissues and organs was low. hTe radioactivity of tumor uptake by using of ROI technology were (88.14±11.59), (123.17±9.06), (98.08±14.40) and (79.87±10.57) at 0.5, 1, 2, 3 h post-injection, and the ratio of T/NT of tumor and liver area were (1.95± 0.19), (3.58±0.78), (3.95±0.39) and (5.01±0.28), respectively.99mTc-EDTA-MN could be quickly cleared from the blood in mice primarily through the kidneys, and the radioactivity in other tissues and organs remained low.Conclusion99mTc-EDTA-MN can be easily prepared and labeled compound with high labeling rate and stability, it appears to be suitable for further experiments requirementin vivo andin vitroapplication.

OBJECTIVETo develop a method for (99m)Tc radiolabeling of a small molecular peptide targeting lung carcinoma and observe the biokinetics and biodistribution of the labeled peptide in normal mice and rabbits.

METHODSMAG3-peptide (cNGQGEQc) was labeled with (99m)Tc and the labeling rate was determined with paper chromatography. In vitro stability test, cysteine challenge test and serum incubation test were performed for radiochemical evaluation of the labeled peptide. Blood (99m)Tc-peptide clearance in rabbits was evaluated by determining blood radioactive concentrations at different time points after injection of 37 MBq (99m)Tc-peptide, and its dynamic distribution was investigated by SPECT imaging. The percent injected dose per gram of tissue was calculated for each organ of mice injected intravenously with 7.4 MBq (99m)Tc-peptide based on gamma counter readings.

RESULTSThe labeling rate of (99m)Tc-peptide exceeded 90%, and the radiochemical purity was 91% after placing for 12 h at room temperature and 85% after incubation at 37 degrees celsius; with human serum. The cysteine replacement rate was less than 7%, and the binding rate of (99m)Tc-peptide with serum proteins was below 5%. SPECT imaging showed that the labeled peptide could be quickly cleared from the blood in normal animals primarily through the kidneys, and the radioactivity in other tissues and organs remained low.

CONCLUSION(99m)Tc-peptide can be easily prepared with a high labeling yield. With good stability both in vitro and in vivo, (99m)Tc-peptide can be quickly cleared from the blood and excreted though the kidney with ideal biodistribution and biokinetics in vivo.

Animals ; Humans ; Male ; Mice ; Molecular Probes ; Organotechnetium Compounds ; blood ; Rabbits ; Tomography, Emission-Computed, Single-Photon ; methods