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*

Objective:To explore the disease spectrum and corresponding clinical indicators of infantile cholestasis so as to provide a basis for the diagnosis of this type of disease at an early stage.Methods:The clinical data was collected from 203 hospitalized children diagnosed with infantile cholestasis at the Department of Gastroenterology of Maternal and Child Health Care, Guiyang City, from January 2018 to March 2023, including 130 males and 73 females. Patients general condition, personal history, and blood biochemical test indicators, including liver and coagulation function, blood ammonia, blood lipid profile, blood sugar, TORCH, thyroid function, and others, were retrospectively analyzed after admission. Cholangiography and high-throughput gene sequencing were performed in certain patients. The etiology of the enrolled cases were analyzed. Children's clinical data were compared with distinct inherited metabolic liver diseases (Group A) and biliary atresia (Group B). The statistical analysis was conducted using the t-test, Mann-Whitney test, Kruskal-Wallis test, or χ2 test, according to different data. Results:In 33 cases, infectious factors—primarily CMV infection—were the etiology of cholestasis. Forty cases had aberrant bile duct development, primarily biliary atresia, choledochal cysts, and intrahepatic bile duct dysplasia. In 26 cases, genetic metabolic factors mainly included citrin protein deficiency, sodium-taurocholate co-transporting polypeptide deficiency, and Alagille syndrome. 11 cases had drug/poisoning factors (parenteral nutrition-associated cholestasis). 19 cases had idiopathic infantile cholestasis. Three cases had other factors; however, all of them had Kawasaki disease. 71 cases had an unclear diagnosis. There was no statistically significant difference in terms of gender and age between groups A and B ( P>0.05). The alkaline phosphatase (ALP) and bile acid levels were significantly higher in Group A than Group B, with a P<0.05, while the gamma glutamyltransferase (GGT), direct bilirubin (DBil), and albumin levels were lower than those in Group B, with a P<0.05. The cytomegalovirus infection rate was higher in Group B (62.50%) than Group A (34.62%), and the difference between the two groups was statistically significant ( χ2=3.89, P<0.05). The alanine aminotransferase, aspartate aminotransferase, GGT, DBil, and albumin were significantly lower in patients with citrin protein deficiency than those in patients with biliary atresia, while ALP, bile acid, and blood ammonia were higher than those in patients with biliary atresia. Patients with sodium-taurocholate co-transporting polypeptide deficiency had higher bile acid than patients with biliary atresia, while the DBil was lower than that in patients with biliary atresia, and the difference was statistically significant ( P<0.05). Conclusion:Infantile cholestasis etiology is diverse. ALP, bile acids, GGT, DBil, and albumin levels can serve as simple indicators for early-stage differentiation between inherited metabolic liver disease and biliary atresia. The cholestasis etiology should be determined as early as possible following biliary atresia exclusion by actively completing genetic metabolic gene detection.

Objective: Based on the information-motivation-behavioral skills (IMB) model theory, the structural equation model(SEM)was used to analyze the effect of using new media to carry out AIDS health education among young students. Methods: Stratified random cluster sampling method was used to select 2 561 students from four colleges in Yantai to conduct a questionnaire survey, and to carry out various forms of new media AIDS health education for the intervention group. Data were analyzed with structural equation model. Results: After intervention，the awareness rate of AIDS were 85.9% and 94.0% respectively(χ2=46.67，P<0.01). The structural equation model fits well, and the fitted indices were: RMSEA=0.04, GFI=0.97, AGFI=0.96, IFI=0.97, CFI=0.98. The direct effects of new media interventions on prevention information and prevention motivation were 0.30 and 0.65 respectively. The influence of information and motivation on preventive behavior was mainly reflected through the indirect effect of behavioral skills. The overall effects of standardization were 0.09 and 0.29 respectively. The overall effect of behavioral skills on preventive behavior was 0.80, which was the main factor affecting HIV prevention behavior. Conclusion The use of new media to carry out AIDS health education among young students had a significant effect. Motivation promotion should be used as a new breakthrough point for future work, and information, motivation, and behavioral skills should be used to promote the prevention of AIDS prevention among young students.

Objective: To investigate the incidence of complications following minimal invasive subcutaneous internal fixator (INFIX) for the treatment of anterior pelvic ring instability, and to analyze control measures. Methods: Data of 42 patients with anterior pelvic ring instability who were treated by anterior subcutaneous internal fixation in our hospital from January 2016 to December 2017 were retrospectively analyzed. There were 15 females and 27 males with an average age of 45.4 years (range, 18-67 years). There were 26 traffic injuries, 10 falling injuries, 3 crush injuries and 3 low energy injuries. According to Tile classification, 24 cases of B2 type, 16 cases of B3 type, 1 case of C1 type and 1 case of C2 type. According to Young-Burgess classification, there were 20 cases of LCI, 18 cases of LCII, 1 case of LCIII, 1 case of APCI, 1 case of APCII, and 1 case of VS. Fourteen cases were combined with limb fractures, and 11 cases were combined with thoracic or abdominal injuries requiring surgical treatment, while 16 cases were with brain injuries. For type APC2, LC1, and some LC2 fractures, fixation for the anterior pelvic ring is enough with INFIX technique alone. For some LC2, LC3, or VS fractures, the anterior and posterior ring were both fixed. Postoperative reduction was evaluated by Matta radiological criteria. Lateral thigh numbness and pain, quadriceps muscle power were recorded during follow-up, and clinical efficacy was evaluated by Majeed score at 6 months after operation. Results: All the 42 patients were followed up for 6 to 12 months, with an average of 9.3 months. Matta standard evaluation for fracture reduction showed that 28 cases were excellent, 14 cases good, and the excellent and good rate was 100%. The early complication rate (within 3 month after operation) was 23.8% (10/42) including 6 cases of lateral femoral cutaneous nerve injury and 2 cases of femoral nerve injury. The symptoms were obviously relieved after the treatment of nutrient nerve and hyperbaric oxygen; 1 case had incision infection which was healed after anti-infection therapy and internal plant removal; 1 case suffered from superior gluteal artery injury which was controlled by pressure hemostasis. Six months after surgery, no case had lost reduction. At 6 months follow-up, the Majeed score was 72-96, with an average of 84.96, of which 32 were excellent and 10 were good, thus the excellent and good rate was 100% (42/42). Conclusion INFIX is an effective internal fixation method for the treatment of unstable anterior pelvic ring injury. However, it has a high rate of early complications, among which nerve injury has the highest incidence.

Objective:To compare the difference of serum levels of M-phospholipaseA2 receptor ( PLA2R) antibodies in patients with idiopathic membranous nephropathy(IMN) detected by two different methods and evaluate the diagnostic value of two methods. Methods:Patients diagnosed as membranous nephropathy and other diseases with biopsy-proven from december 2014 to october 2015 in Shengjing Hospital of China Medical University were enrolled and devided into IMN group and non-IMN group. The serum levels of anti-PLA2R antibody were detected by both indirect immunofluorescence assay(IFA) and enzyme linked immunosorbent assay(ELISA). Results:The sensitivity of IFA and ELISA in IMN were 71. 3% and 68. 5%,and the specificities of two methods were the same as 100%. The area under ROC curves of anti-PLA2R antibody for IMN diagnosis were 0. 860 and 0. 839. The diagnostic value of IFA and ELISA was no statistically significant differences in IMN ( P>0. 05 ) , and the consistency of two methods was better (κ=0. 876). The IMN patients of positive anti-PLA2R antibody be susceptible to the low level of serum albumin (P<0. 05). The higher levels of PLA2R antibody were linked with the worse hypoproteinemia and the higher rate of nephrotic-range proteinuria in IMN patients. Conclusion:Two methods of detecting sera PLA2R antibody have higher sensitivity and specificity,so the sera anti-PLA2R antibody was a better biomarker in the diagnosis of idiopathic membranous nephropathy.

Objective To investigate therapeutic effects of β-TCP/PLGA scaffolds seeded bone marrow mesenchymal stem cells (BMSCs) combing with hyperbaric oxygen (HBO) on bone defect with seawater immersing in rabbit radius.Methods BMSCs were seeded into the β-TCP/PLGA scaffolds to construct tissue engineering bone.60 New Zealand rabbits were created 1.5 cm bone defect in bilateral radius,and then bilateral wound limbs were immersed in seawater for 3 h.After debridement,all rabbits were randomly divided into 4 groups.In group A nothing was implanted into the bone defect.Only BMSCs was implanted in group B.BMSCs + HBO was given in group C.β-TCP/PLGA BMSCs + HBO was given in group D.At postoperative 4,8,12 weeks rabbits were sacrificed after radiography radius.X-ray radiography,HE staining,immunohistochemical observation were used to evaluate repairing effect of bone defect with seawater immersing.Results Radiographic analysis demonstrated that the bone defects were completely repaired with recanalization of the medullary cavity in group D,bone defects were partially repaired with partal recanalization of the medullary cavity in group C,bone defects were incompletely repaired in group B;bone defect repair unfinished and the broken ends were sclerous in group A.Each time point group D＞group C＞group B＞group A in callus grey value (P＜0.05).HE staining indicated after 12 weeks,a small amount of lainellar bone formated in group A;a small amount of lamellar bone formated in group B;a large amount of lamellar bone formated in group C;a large amount of lamellar bone formated in group D.Immunohistoehemical result indicated at postoperative 4 weeks,the expressions of osteocalcin(OCN) in each group were at high levels,the expressions decreased significantly at Postoperative 8 weeks,the expressions were at a low level at postoperative 12 weeks.At postoperative 4 and 8 weeks,the expression levels of OCN were group D＞ group C＞ group B＞group A (P＜0.05),there was no significant difference at postoperative 12 weeks between the 4 groups (P＞0.05).Conelusion the β-TCP/PLGA composite BMSCs combined hyperbaric oxygen is the effective method to repair seawater immersed bone defect in rabbit.

Objective To investigate the effects of HBO on open tibial fracture coupled with seawater immersion in rabbits and the possible mechanism involved.Methods Forty-eight New Zealand rabbits were selected and randomly divided into the HBO group and the control group,each consisting of 24 animals.The animals of both groups were inflicted with unilateral open tibial fracture.The injured limbs were immersed in artificial seawater for 3 hours,and then had wound suture and internal fracture fixation after debridement.The animals in the HBO group was immediately treated with HBO,once a day for a succession of 2 weeks,whereas the animals in the Control group were left there untreated.At week 4,8 and 12 after injury,the animals were sacrificed and samples (8 limbs from each group) were collected for further study.The repair of the fractured tissues of both groups was compared by radiography,histology and immunohistochemistry.Results (1)Histopathology was performed by HE staining.At week 12 after injury,for the animals in the HBO group,osseous callus could universally be seen in the fracture zone,cell components were basically mature bone cells,and the infiltration of inflammatory cells could scarcely be seen,while for the animals in the Control group,mixed bone callus could mainly be noted in the fracture zone,but cartilage cells were still visible,and a small scale of inflammatory cell infiltration could also be observed.(2) Immunohistochemistry revealed that vascular endothelial growth factor (VEGF) was mainly expressed in the cytoplasm of osteoblast,chondrocytes and osteocyte,and significant changes could be seen at different time points.At week 4 after injury,the expression levels of the two groups were the highest.At week 8 and 12 after injury,the expression levels all gradually dropped to a relatively low level.At week 4 and 8 after injury,the expression level of the HBO group was obviously stronger than that of the Control group(P ＜0.01).However,at week 12 after injury,no significant differences could be noted in the expression levels between the 2 groups (P ＞ 0.05).(3) Radiographic imaging was performed for the detection of callus grayscale in the fracture zone.At week 12 after injury,the average value of callus grayscale in the fracture zone for the animals of the HBO group was (190.58 ± 7.52),while that of the Control group was (144 ± 8.11).Statistically significance was shown,when comparisons were made between the 2 groups (P ＜0.01).(4)At week 4 after injury,the content of malondialdehyde (MDA) in the soft tissue around the fracture zone in the animals of the HBO group was significantly lower than that of the Control group,and the activity of superoxide dismutase (SOD) in the animals of the HBO group was higher than that of the Control group.Statistical significance could be seen,when comparisons were made between the 2 groups (P ＜ 0.01).However,at week 8 and 12 after injury,there were no significant differences in the content of MDA and activity of SOD between the 2 groups (P ＞ 0.05).Conclusions In the treatment of open bone fracture coupled with seawater immersion,HBO could alleviate inflammatory reaction and reperfusion injury,enhance the proliferation of tissue cells,and was beneficial to the healing of bone fracture.For this reason,this regimen was a practical method for the repair of open bone fracture coupled with seawater immersion.

Objective To investigate the effects of HBO on open tibial fracture coupled with seawater immersion in rabbits and the possible mechanism involved.Methods Forty-eight New Zealand rabbits were selected and randomly divided into the HBO group and the control group,each consisting of 24 animals.The animals of both groups were inflicted with unilateral open tibial fracture.The injured limbs were immersed in artificial seawater for 3 hours,and then had wound suture and internal fracture fixation after debridement.The animals in the HBO group was immediately treated with HBO,once a day for a succession of 2 weeks,whereas the animals in the Control group were left there untreated.At week 4,8 and 12 after injury,the animals were sacrificed and samples (8 limbs from each group) were collected for further study.The repair of the fractured tissues of both groups was compared by radiography,histology and immunohistochemistry.Results (1)Histopathology was performed by HE staining.At week 12 after injury,for the animals in the HBO group,osseous callus could universally be seen in the fracture zone,cell components were basically mature bone cells,and the infiltration of inflammatory cells could scarcely be seen,while for the animals in the Control group,mixed bone callus could mainly be noted in the fracture zone,but cartilage cells were still visible,and a small scale of inflammatory cell infiltration could also be observed.(2) Immunohistochemistry revealed that vascular endothelial growth factor (VEGF) was mainly expressed in the cytoplasm of osteoblast,chondrocytes and osteocyte,and significant changes could be seen at different time points.At week 4 after injury,the expression levels of the two groups were the highest.At week 8 and 12 after injury,the expression levels all gradually dropped to a relatively low level.At week 4 and 8 after injury,the expression level of the HBO group was obviously stronger than that of the Control group(P ＜0.01).However,at week 12 after injury,no significant differences could be noted in the expression levels between the 2 groups (P ＞ 0.05).(3) Radiographic imaging was performed for the detection of callus grayscale in the fracture zone.At week 12 after injury,the average value of callus grayscale in the fracture zone for the animals of the HBO group was (190.58 ± 7.52),while that of the Control group was (144 ± 8.11).Statistically significance was shown,when comparisons were made between the 2 groups (P ＜0.01).(4)At week 4 after injury,the content of malondialdehyde (MDA) in the soft tissue around the fracture zone in the animals of the HBO group was significantly lower than that of the Control group,and the activity of superoxide dismutase (SOD) in the animals of the HBO group was higher than that of the Control group.Statistical significance could be seen,when comparisons were made between the 2 groups (P ＜ 0.01).However,at week 8 and 12 after injury,there were no significant differences in the content of MDA and activity of SOD between the 2 groups (P ＞ 0.05).Conclusions In the treatment of open bone fracture coupled with seawater immersion,HBO could alleviate inflammatory reaction and reperfusion injury,enhance the proliferation of tissue cells,and was beneficial to the healing of bone fracture.For this reason,this regimen was a practical method for the repair of open bone fracture coupled with seawater immersion.

Objective To observe the effects of Yushen Heji on expressions of Heme Oxygenase (HO-1) and HO-2 in kidney of diabetic rats;To explore its mechanism in diabetic nephropathy. Methods SD rats were randomly divided into normal group, model group, control group and treatment group. Models were duplicated by subcutaneous injection STZ. Control group and treatment group were given gavage with related medicine, while normal group and model group were given gavage with the same amount of distilled water. Eight weeks later, the urine of rats was collected to detect the 24 h Upro and Ucr;the rats were sacrificed to detect the levels of CO, BUN and Scr;the kidneys were harvested for observing the expressions of HO-1 and HO-2 by immunohistochemistry.Results Compared with normal group, the contents of BUN, 24 h Upro and Scr all increased in model group;Ccr and plasma CO decreased;the expressions of HO-1 and HO-2 in kidney tissues were obviously reduced (P<0.01). Compared with model group, the contents of BUN, 24 h Upro and Scr all obviously decreased in treatment group and control group;Ccr and plasma CO increased;the expressions of HO-1 and HO-2 in kidney tissues obviously increased (P<0.05,P<0.01). Compared with control group, the contents of 24 h Upro decreased in treatment group;plasma CO increased;the expressions of HO-1 and HO-2 in kidney tissues obviously increased (P<0.05,P<0.01).Conclusion Yushen Heji has therapeutic effect on diabetic nephropathy rats, which may related with HO/CO system.

Objective To explore the effects of HBO combined with percutaneous allogeneic bone marrow mesenchymal stem cells (BMSCs) grafting on the repair of rabbit radius open bone defect following seawater immersion.Methods Forty-eight New Zealand rabbits were randomly divided into 4 groups:the blank control group (group A),the(BMSCs) grafting group (group B),the HBO group (group C),and the HBO combined with BMSCs group (group D).A fifteen-millimeter bone defect was made in the mid-section of radius and then was immersed in the artificial seawater for 3 hours,then the wound was closed with debridement.The animals received HBO therapy the moment after completion of debridement.BMSCs grafting was administered 72 hours after debridement.The animals were sacrificed at week 4,8 and 12 to collect the samples (there were 4 rabbits from each batch of the group,and there were altogether 8 limbs).Repairs of the defect areas were compared between groups by means of radiographic scanning,histologic and immunohistochemical analyses.Results HE staining revealed that osteogenesis in the animals of group D at week 4 and 8 was the best,and group C was superior to group B.In the animals of group A,bone repair was not detected in the defect area,which was filled with scar tissue.At week 12 after injury,osteogenesis in the defect areas for the animals of group D was the best,which was mostly mature bone tissue.(2)Immunohistochemical detection indicated that VEGF and TGF-β1 were mostly expressed in the osteoblastic cells,cartilage cells and bone cell cytoplasm,and significant changes could be noted at different time points.At week 4 after injury,the expressions of VEGF and TGF-β1 all reached peak levels.At week 8,their expressions all decreased significantly,as compared with those at week 4 and 12,their expressions all decreased further to a low level.At week 4 and 8 after injury,the expression levels of VEGF and TGF-β1 for the 4 groups were in the following order:group D ＞ group C ＞ group B ＞ group A.Statistical significance could be seen,when comparisons were made between the groups (P ＜0.05).However,at week 12,no significant differences could detected when comparisons were made between the 4 groups (P ＞ 0.05).(3) Osteotylus measurement revealed that osteotylus grayness for the animals of group D,group C,group B and group A at week 12 after injury was 211.84 ± 4.31,200.31 ± 3.08,152.79 ± 4.43 and 80.88 ± 3.31 respectively,and statistical significance could be noted,when comparisons were made between the groups (P ＜ 0.05).Conclusions The effects of simple percutaneous allogeneic bone marrow mesenchymal stem cells grafting or simple HBO therapy on the repair of open bone defect following seawater immersion were not ideal.However,combined applications of these 2 treatment methods could produce synergistic effects,which could promote osteogenetic process of the affected bone and was an applicable method for the repair of open bone defect following seawater immersion.