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*

Objectives:To evaluate the role of pancreas multidisciplinary team(MDT) clinic in the diagnosis of pancreatic diseases,patient compliance with MDT advice,and the impact of MDT on the postoperative survival of patients with pancreatic cancer.Methods:The study included 927 patients(554 males,373 females,aged (58.1±13.3)years (range: 15 to 89 years)) that had visited the pancreas MDT clinic of Zhongshan Hospital from May 2015 to December 2021,and 677 patients(396 males, 281 females, aged (63.6±8.9)years(range: 32 to 95 years)) who underwent radical surgery and with pathologically confirmed pancreatic adenocarcinoma from January 2012 to December 2020,of whom 79 patients had attended the pancreas MDT. The clinical and pathological data were collected and analyzed retrospectively. Diseases were classified in accordance with 2010 WHO classification of tumors of the digestive system and usual clinical practices. The Kaplan-Meier method was used for drawing the survival curve and calculating the survival rate. The univariate analysis was done by Log-rank test and the multivariate analysis was done by COX proportional hazards model. Survival rates were compared using χ 2 test. Results:Among the 927 patients that had visited the MDT clinic,233 patients(25.1%) were referred due to undetermined diagnosis. A direct diagnosis was made in 109 cases (46.8%,109/233) by the MDT clinic, of which 98 were consistent with the final diagnosis,resulting in an accuracy of 89.9%(98/109). The direct diagnosis rate in the recent years(36.6%(41/112),from June 2019 to December 2021) decreased compared to that in the previous years(56.2%(68/121),from May 2015 to May 2019),yet the accuracy in the recent years(90.2%,37/41) was basically the same as before (89.7%,61/68). The rate of compliance of the entire cohort was 71.5%(663/927), with the compliance rate in the recent two and a half years(81.4%,338/415) remarkably higher than that in the previous four years(63.4%,325/512). Patients with pancreatic cancer that attended the MDT exhibited a trend toward longer median postoperative survival than patients that did not attend the MDT,but the difference was not statistically significant(35.2 months vs.30.2 months, P>0.05). The 1-year and 3-year survival rates of patients that attended the MDT were significanly higher than patients that did not attend the MDT(88.6% vs. 78.4%, P<0.05;32.9% vs. 21.9%, P<0.05,respectively),but the 5-year survival rate was not statistically different(7.6% vs. 4.8%, P>0.05). Conclusions:The pancreas MDT clinic is an accurate and convenient way to diagnose intractable pancreatic diseases,and in the recent years the patients′ compliance rate with MDT advice has increased. Pancreatic cancer patients that have attended the MDT have higher 1-year and 3-year postoperative survival rates,but the long-term survival benefits of MDT still needs to be proved by clinical studies on a larger scale.

Objective:To explore the occurrence and characteristics of adverse reactions related to bevacizumab (Bev) in treatment of non-small cell lung cancer (NSCLC).Methods:The medical records of NSCLC patients treated with Bev in the Respiratory Department of Bozhou Hospital of Traditional Chinese Medicine from May 2019 to March 2021 were collected. Patients with Bev-related adverse reactions, which were judged by Naranjo scoring method, were selected. The relevant information in these patients was extracted from their medical records, and the occurrence and outcomes of the adverse reactions (incidence, time of occurrence, clinical manifestation, severity, etc.) were analyzed retrospectively. The dosing regimen of Bev was IV infusion of 7.5 mg/kg once per 21 days (1 cycle).Results:A total of 142 patients were included in the analysis, and 17 (12.0%), 20 (14.1%), 51 (35.9%), 35 (24.6%), and 19 (13.4%) patients received 1, 2, 3, 4, and 5 cycles of Bev treatment, respectively. All patients were treated with combination regimen with traditional chemotherapy drugs, including pemetrexed in 10 patients (7.0%), pemetrexed+carboplatin in 41 patients (28.9%), and pemetrexed+cisplatin in 91 (64.1%) patients, respectively. Among the 142 patients, 49 (34.5%) had adverse reactions, which were classified as grade 1, 2, 3, and 4 in severity in 18, 19, 6, 6 patients respectively, and the incidence of serious adverse reactions (≥ grade 3) was 8.5% (12/142). The clinical manifestations of adverse reactions included hematologic injury in 12 patients (8.5%; grade 1 in 4 and grade 2 in 8 patients), hypertension in 11 patients (7.7%; grade 1 in 4, grade 2 in 3, grade 3 in 2, and grade 4 in 2 patients), skin injury in 8 patients (5.6%; grade 2 in 5 and grade 3 in 3 patients), bleeding events in 6 patients (4.2%; grade 1 in 3, grade 3 in 1, and grade 4 in 2 patients), gastrointestinal reaction in 6 patients (4.2%; grade 1 in 3 and grade 2 in 3 patients), proteinuria in 3 patients (2.1%, grade 1), pulmonary embolism in 1 patient (0.7%, grade 4), gastric perforation in 1 patient (0.7%, grade 4), and alopecia in 1 patient (0.7%, grade 1). Bev was not discontinued in 37 patients who developed grade 1-2 adverse events, and 25 of them were given symptomatic treatments. Patients with grade ≥3 adverse reactions stopped Bev and received symptomatic treatments. All patients recovered or were improved.Conclusions:The common adverse reactions in Bev treatment for NSCLC include hematologic injury, hypertension, skin injury, bleeding events, gastrointestinal reaction, and proteinuria, most of which are of grade 1-2 in severity. Severe adverse effects such as pulmonary embolism and gastric perforation occurred occasionally. Patients with grade ≥ 3 adverse reactions have a good prognosis when Bev is discontinued and symptomatic treatments are given.

Objective:To investigate the clinical effect of drugs combined with hyperbaric oxygen（HBO）on nervous system damage caused by nitrous oxide（N 2O）poisoning. Methods:The clinical data of 12 patients with nerve damage caused by N 2O poisoning admitted to the Department of Neurology，Hyperbaric Oxygen of a certain PLA hospital from February 2020 to April 2021 were reviewed. The changes of neuroelectrophysiological and neurological functions of patients before and after treatment were analyzed using electroneurogram detection，Berg balance scale，and Holden’s functional ambulation categories（FAC）as observation indicators. Results:After drug treatment combined with HBO，the motor conduction velocity，sensory conduction velocity，motor latency，motor evoked potential，and sensory evoked potential of electroneurogram of 12 patients were significantly improved，and the differences were statistically significant（ P < 0.05）.Berg balance scale scores were significantly improved compared with those before treatment，and the differences were statistically significant（ P < 0.05）.FAC scale was improved compared with before treatment，but the difference was not statistically significant（ P > 0.05）. Conclusion:HBO combined with folic acid，vitamin B12 and other neurotrophic drugs has a certain clinical effect in the treatment of nerve damage caused by N 2O poisoning，which is worthy of further study and improvement.

Objectives:To evaluate the role of pancreas multidisciplinary team(MDT) clinic in the diagnosis of pancreatic diseases,patient compliance with MDT advice,and the impact of MDT on the postoperative survival of patients with pancreatic cancer.Methods:The study included 927 patients(554 males,373 females,aged (58.1±13.3)years (range: 15 to 89 years)) that had visited the pancreas MDT clinic of Zhongshan Hospital from May 2015 to December 2021,and 677 patients(396 males, 281 females, aged (63.6±8.9)years(range: 32 to 95 years)) who underwent radical surgery and with pathologically confirmed pancreatic adenocarcinoma from January 2012 to December 2020,of whom 79 patients had attended the pancreas MDT. The clinical and pathological data were collected and analyzed retrospectively. Diseases were classified in accordance with 2010 WHO classification of tumors of the digestive system and usual clinical practices. The Kaplan-Meier method was used for drawing the survival curve and calculating the survival rate. The univariate analysis was done by Log-rank test and the multivariate analysis was done by COX proportional hazards model. Survival rates were compared using χ 2 test. Results:Among the 927 patients that had visited the MDT clinic,233 patients(25.1%) were referred due to undetermined diagnosis. A direct diagnosis was made in 109 cases (46.8%,109/233) by the MDT clinic, of which 98 were consistent with the final diagnosis,resulting in an accuracy of 89.9%(98/109). The direct diagnosis rate in the recent years(36.6%(41/112),from June 2019 to December 2021) decreased compared to that in the previous years(56.2%(68/121),from May 2015 to May 2019),yet the accuracy in the recent years(90.2%,37/41) was basically the same as before (89.7%,61/68). The rate of compliance of the entire cohort was 71.5%(663/927), with the compliance rate in the recent two and a half years(81.4%,338/415) remarkably higher than that in the previous four years(63.4%,325/512). Patients with pancreatic cancer that attended the MDT exhibited a trend toward longer median postoperative survival than patients that did not attend the MDT,but the difference was not statistically significant(35.2 months vs.30.2 months, P>0.05). The 1-year and 3-year survival rates of patients that attended the MDT were significanly higher than patients that did not attend the MDT(88.6% vs. 78.4%, P<0.05;32.9% vs. 21.9%, P<0.05,respectively),but the 5-year survival rate was not statistically different(7.6% vs. 4.8%, P>0.05). Conclusions:The pancreas MDT clinic is an accurate and convenient way to diagnose intractable pancreatic diseases,and in the recent years the patients′ compliance rate with MDT advice has increased. Pancreatic cancer patients that have attended the MDT have higher 1-year and 3-year postoperative survival rates,but the long-term survival benefits of MDT still needs to be proved by clinical studies on a larger scale.

Objective:To explore the occurrence and characteristics of adverse reactions related to bevacizumab (Bev) in treatment of non-small cell lung cancer (NSCLC).Methods:The medical records of NSCLC patients treated with Bev in the Respiratory Department of Bozhou Hospital of Traditional Chinese Medicine from May 2019 to March 2021 were collected. Patients with Bev-related adverse reactions, which were judged by Naranjo scoring method, were selected. The relevant information in these patients was extracted from their medical records, and the occurrence and outcomes of the adverse reactions (incidence, time of occurrence, clinical manifestation, severity, etc.) were analyzed retrospectively. The dosing regimen of Bev was IV infusion of 7.5 mg/kg once per 21 days (1 cycle).Results:A total of 142 patients were included in the analysis, and 17 (12.0%), 20 (14.1%), 51 (35.9%), 35 (24.6%), and 19 (13.4%) patients received 1, 2, 3, 4, and 5 cycles of Bev treatment, respectively. All patients were treated with combination regimen with traditional chemotherapy drugs, including pemetrexed in 10 patients (7.0%), pemetrexed+carboplatin in 41 patients (28.9%), and pemetrexed+cisplatin in 91 (64.1%) patients, respectively. Among the 142 patients, 49 (34.5%) had adverse reactions, which were classified as grade 1, 2, 3, and 4 in severity in 18, 19, 6, 6 patients respectively, and the incidence of serious adverse reactions (≥ grade 3) was 8.5% (12/142). The clinical manifestations of adverse reactions included hematologic injury in 12 patients (8.5%; grade 1 in 4 and grade 2 in 8 patients), hypertension in 11 patients (7.7%; grade 1 in 4, grade 2 in 3, grade 3 in 2, and grade 4 in 2 patients), skin injury in 8 patients (5.6%; grade 2 in 5 and grade 3 in 3 patients), bleeding events in 6 patients (4.2%; grade 1 in 3, grade 3 in 1, and grade 4 in 2 patients), gastrointestinal reaction in 6 patients (4.2%; grade 1 in 3 and grade 2 in 3 patients), proteinuria in 3 patients (2.1%, grade 1), pulmonary embolism in 1 patient (0.7%, grade 4), gastric perforation in 1 patient (0.7%, grade 4), and alopecia in 1 patient (0.7%, grade 1). Bev was not discontinued in 37 patients who developed grade 1-2 adverse events, and 25 of them were given symptomatic treatments. Patients with grade ≥3 adverse reactions stopped Bev and received symptomatic treatments. All patients recovered or were improved.Conclusions:The common adverse reactions in Bev treatment for NSCLC include hematologic injury, hypertension, skin injury, bleeding events, gastrointestinal reaction, and proteinuria, most of which are of grade 1-2 in severity. Severe adverse effects such as pulmonary embolism and gastric perforation occurred occasionally. Patients with grade ≥ 3 adverse reactions have a good prognosis when Bev is discontinued and symptomatic treatments are given.

Objective:To investigate the clinical effect of drugs combined with hyperbaric oxygen（HBO）on nervous system damage caused by nitrous oxide（N 2O）poisoning. Methods:The clinical data of 12 patients with nerve damage caused by N 2O poisoning admitted to the Department of Neurology，Hyperbaric Oxygen of a certain PLA hospital from February 2020 to April 2021 were reviewed. The changes of neuroelectrophysiological and neurological functions of patients before and after treatment were analyzed using electroneurogram detection，Berg balance scale，and Holden’s functional ambulation categories（FAC）as observation indicators. Results:After drug treatment combined with HBO，the motor conduction velocity，sensory conduction velocity，motor latency，motor evoked potential，and sensory evoked potential of electroneurogram of 12 patients were significantly improved，and the differences were statistically significant（ P < 0.05）.Berg balance scale scores were significantly improved compared with those before treatment，and the differences were statistically significant（ P < 0.05）.FAC scale was improved compared with before treatment，but the difference was not statistically significant（ P > 0.05）. Conclusion:HBO combined with folic acid，vitamin B12 and other neurotrophic drugs has a certain clinical effect in the treatment of nerve damage caused by N 2O poisoning，which is worthy of further study and improvement.

Objective:To discuss the effect of modified ultrafiltration combined with sequential infusion of blood products, such as platelets and cryoprecipitation, on perioperative coagulation function in neonates undergoing cardiovascular surgery under extracorporeal circulation.Methods:A retrospective analysis was performed on 83 neonates who underwent cardiovascular surgery from January 2017 to December 2019. They were divided into the control group (conventional ultrafiltration, 51 cases) and the treatment group (modified ultrafiltration + sequential infusion of blood products, 32 cases).Results:The age of treatment group was significantly younger than that of the control group ( P<0.05). The extracorporeal circulation time of the treatment group was significantly longer than that of the control group. Compared with the preoperative data, post-APTT, post-PT and post-INR were increased significantly in the control group, platelet count and FIB were decreased significantly ( P<0.01), while there was no significant difference in the treatment group. Compared with the treatment groups, post-APTT, post-PT and post-INR were significantly increased in the control group, and postoperative platelet count and FIB were significantly decreased ( P<0.05). The drainage of control group was significantly higher than that of the treatment group at 12 h and 24 h after surgery ( P<0.01). Conclusion:The application of modified ultrafiltration combined with sequential infusion of blood products can significantly improve perioperative coagulation and reduce mediastinal bleeding in neonatal after cardiovascular surgery.

To alleviate the shortage of excellent talents of traditional Chinese medicine (TCM), the State Administration of Traditional Chinese Medicine has decided to cultivate a group of talents with strong theoretical foundation, rich practical experience and exquisite skills who would like to devote themselves to TCM technology research. The national talent training program for inheritance of characteristic TCM technology is a major breakthrough in the history of new China in the cultivation of high-level TCM technology talents. A retrospective analysis was performed on the national talent training program for inheritance of characteristic TCM technology hold in training bases from Beijing, Shanghai, Guangdong Province, Jiangsu Province, Anhui Province, Xinjiang Uygur Autonomous Region, Inner Mongolia Autonomous Region, Heilongjiang Province, Yunnan Province, Sichuan Province, Shaanxi Province and Fujian Province. Based on the completion assessment work of 2014 and 2015 national talent training program for inheritance of characteristic TCM technology, and after knowledge exchange and discussion with the training subjects from 30 provinces and some teaching experts in the field of characteristic TCM technology, this paper summarizes the work of the national talent training program for inheritance of characteristic TCM technology. After trained in bases in China, training subjects' theoretical knowledge and practical skills as well as their comprehensive capacity have been greatly promoted. At the same time, there are also some problems and shortages. For example, characteristic TCM technology in some training bases is not prominent enough, some characteristic TCM technologies are not well inherited, and the affiliations of some training subjects do not pay enough attention to it. All of these problems should be solved and improved in future work.

Objective:To review and analyze the clinical experience of children with aortic valve stenosis and/or insufficiency treated with autologous pulmonary valve for aortic valve replacement procedure(Ross operation) with ePTFE artificial valve for right ventricular outflow tract reconstruction.Methods:From 2015 to 2020, 8 cases of aortic stenosis and/or aortic insufficiency treated by Ross operation in our center were collected, with an age of 0.5-13.2 years old. 4 cases of aortic stenosis were diagnosed preoperatively, 3 cases of aortic stenosis with aortic insufficiency, and 1 case of infective endocarditis involving the aortic valve. The operation was carried out in three steps: Harvest autologous pulmonary valve; the diseased aortic valve was resected and autologous pulmonary valve was transplanted to the aorta by aortic root transplantation; the right ventricular outflow tract was reconstructed by a handmade ePTFE artificial flap blood vessel.Results:In 6 cases, the right ventricular outflow tract was reconstructed by hand-sewn ePTFE trileaflets, and artificial univalve in 2 cases, no death occurred during operation; all patients were cured and discharged. The patients were followed up for 1 to 36 months, with mean of(12.63±12.19) months. There was no long-term death or valvular complications. During follow-up echocardiography indicated 1 case of moderate aortic regurgitation, 1 case of mild-moderate regurgitation, and moderate regurgitation was found in 2 patients with artificial single pulmonary valve. For the remaining patients, they were mild aortic regurgitation, and a trivial or mild pulmonary artery regurgitation with hand-sewn three-leaflets ePTFE artificial vessel; All patients were followed up at the last time with a peak pressure of(6.63±3.46) mmHg(1 mmHg=0.133 kPa) across the aortic valve. The left ventricular outflow tract and aortic annulus shrank slightly after surgery(the diameter of one patient with Ross-Konno operation increased), but the annulus diameter increased with age. There was no need for further intervention.Conclusion:The Ross operation is safe for the treatment of aortic valve disease, it has good hemodynamic effect, and the autologous pulmonary artery has growth potential, especially suitable for children and young patients. Hand-sewn ePTFE with trileaflet vessels for reconstruction of right ventricular outflow tract performs well in anti-regurgitation function in the short term or may be used as a replacement material for the homograft/heterograft vessel, but longer follow-up and more cases are needed.