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*

Osteoporotic vertebral compression fracture (OVCF) can lead to lower back pain and may be even accompanied by scoliosis, neurological dysfunction and other complications, which will affect the daily activities and life quality of patients. Vertebral augmentation is an effective treatment method for OVCF, but it cannot correct unbalance of bone metabolism or improve the osteoporotic status, causing complications like lower back pain, limited spinal activities and vertebral refracture. The post-operative systematic and standardized rehabilitation treatments can improve curative effect and therapeutic efficacy of anti-osteoporosis, reduce risk of vertebral refracture, increase patient compliance and improve quality of life. Since there still lack relevant clinical treatment guidelines for postoperative rehabilitation treatments following vertebral augmentation for OVCF, the current treatments are varied with uneven therapeutic effect. In order to standardize the postoperative rehabilitation treatment, the Spine Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized relevant experts to refer to relevant literature and develop the "Guideline for postoperative rehabilitation treatment following vertebral augmentation for osteoporotic vertebral compression fracture (2022 version)" based on the clinical guidelines published by the American Academy of Orthopedic Surgeons (AAOS) as well as on the principles of scientificity, practicality and advancement. The guideline provided evidence-based recommendations on 10 important issues related to postoperative rehabilitation treatments of OVCF.

Objective:To investigate the surgical strategy of posterior correction of cervicothoracic scoliosis in children and adolescents, and to analyze the curative effect of surgical correction.Methods:A retrospective study was conducted on 14 patients with cervicothoracic scoliosis who underwent surgical treatment in the department of spine surgery of our hospital from January 2014 to June 2020, including 9 female and 5 male patients. 8 patients were treated with Halo traction before surgery.Among them, 7 patients were treated by posterior column osteotomy and fusion surgery, 7 patients were treated byposterior approach hemivertebra osteotomy. The scoliosis Cobb angle, T 1 tilt angle, clavicle angle, neck tilt angle, shoulder height difference, sagittal balance distance, coronal balance distance and local kyphosis angle were measured compared among before operation, after operation, at 1 year follow-up and at the last follow-up to evaluate the effect of surgical treatment and the correction loss at follow-up. Intraoperative and postoperative complications were recorded, and the Scoliosis Research Society question naires-22 (SRS-22) questionnaire was completed preoperatively and at 24-month follow-up to evaluate the functional status and treatment effect. Results:All 14 patients successfully completed the operation, the operation time was 6.85±1.79 h (range, 5-11 h); the intraoperative blood loss was 685.71±265.61 ml (range, 400-1 200 ml), and the follow-up time was 37.28±13.75 months (range, 24-72 months). The Cobb angle of the main curve was 50.20°±15.19° preoperatively, 10.91°±6.46° postoperatively , 10.53°±6.42° at 1-year follow-up, and 10.14°±5.95° at the last follow-up, and the difference was statistically significant ( F=45.55, P<0.001), the preoperative and postoperative difference was statistically significant ( t=10.62, P<0.001) with a correction rate of 78.32%±11.41%. The T 1 inclination angle was 16.08°±8.06° before operation, 3.71°±2.40° after operation, 4.05°±1.94° at 1-year follow-up, and 3.97°±2.04° at the last follow-up, and the difference was statistically significant ( F=10.55, P=0.001), the preoperative and postoperative difference was statistically significant ( t=6.37, P<0.001) with a correction rate of 69.56%±25.86%. The neck tilt angle was 7.45°±3.72° before operation, 2.45°±1.12° after operation, 2.75°±0.89° at 1-year follow-up, and 3.10°±2.01° at the last follow-up, and the difference was statistically significant ( F=6.65, P=0.008), in which postoperative correction rate was 57.92%±25.41%, and the difference was statistically significant ( t=4.69, P<0.001). The data of shoulder height difference before operation did not conform to normal distribution (Shapiro-Wilk test, P=0.017), it was 0.97 (0.54, 1.32) cm before operation and 0.53±0.40 cm after operation, and the postoperative correction rate was 50.17%±27.38%, the difference was statistically significant ( Z=3.18, P=0.001). The total score of SRS-22 questionnaire was increased from 4.21±0.29 preoperatively to 4.81±0.17 at 24-month follow-up ( t=7.35, P<0.001). Except for one patient with transient upper limb numbness, the other 13 patients showed no obvious intraoperative or postoperative complications. Conclusion:Both posterior column osteotomy with fusion and posterior hemivertebra osteotomy are effective in the treatment of cervicothoracic scoliosis, and the surgeon can make individual treatment plans according to different conditions.

Objective To explore the clinical effect of neoadjuvant chemotherapy combined with tumor cell subtraction in the treatment of advanced epithelial ovarian cancer,as well as the effects on serum epididymal secretory protein 4 ( Human Epididymis protein 4,HE4) ,and glucose polypeptide antigen 125 ( cancer antigen 125,CA125 ) . Methods From January 2010 to January 2014, patients with advanced ovarian cancer from Quanzhou First Hospital Affiliated to Fujian Medical University were selected. According to the difference of clinical treatment plan,128 patients with advanced epithelial ovarian cancer were divided into the observation group ( 68 cases ) and the control group ( 60 cases ) . The patients in the observation group were given neoadjuvant chemotherapy combined with tumor cytoreductivesurgery, and the control group was treated with tumor cytoreductivesurgery and conventional chemotherapy. The clinical efficacy, operation time, blood loss volume, ascites volume, complication, hospitalization time, HE4and CA125 were compared between the two groups. Also 1 year,3 year survival rate,HE4 and CA125 levels of the two groups were analyzed. Results The number of satisfactory tumor reduction cases in observation group was significantly higher than that in control group( 73. 53％( 50/68) ,51. 67％( 31/60) ,χ2=6. 56,P<0. 05) . The short-term effect of observation group was significantly better than that in control group( Z=5. 79,P<0. 05) . The operation time,blood loss volume,ascites volume,complication and hospitalization time of observation group were significantly lower than those in control group (operation time:(119. 6±39. 1) min vs. (177. 3±45. 6) min,t=7. 71,P<0. 05;blood loss:(378. 9 ±88.4) ml vs. (616.3±110.8) ml,t=13.47,P<0.05;ascites volume:(678.5±205.1) ml vs. (1372.4 ±405. 8) ml,t=12. 42, P<0. 05;complication: 13. 2％( 9/68) vs. 31. 7％( 19/60),χ2 = 6. 34, P<0. 05;hospitalization time:(10. 4±3. 2)d vs. (15. 3±3. 1)d,t=8. 77,P<0. 05). There was no significant difference in HE4 and CA125 between the two groups before treatment ( P>0. 05) . After chemotherapy,the level of HE4 and CA125 decreased significantly in the two groups after chemotherapy,and the observation group was significantly lower than the control group (HE4: (98. 3±28. 9) pmol/L vs. (153. 2±44. 1) pmol/L,t=8. 42,P<0. 05;CA125:(35. 3±14. 8) vs. (48. 3±14. 2) ) kU/L,t=5. 05,P<0. 05). The myelosuppression and digestive tract reaction in the observation group were more serious than those in control group (χ2=4. 09,4. 87,P<0. 05) . There was no significant difference in the 1 year survival rate between the two groups ( 86. 76％ vs. 81. 67％,χ2=0. 63, P>0. 05 ) . After 1 years of follow-up, the levels of HE4 and CAl25 in the observation group were significantly lower than those in the control group (HE4:(112. 2±33. 7) pmol/L vs. (189. 4±39. 6) pmol/L,t=10. 95,P<0. 05;CA125:(51. 2±14. 2) kU/L vs. (59. 7±18. 6) kU/L,t=2. 69,P<0. 05). 3 year survival rate in observation group was significantly higher than that in control group ( 55. 88％ vs. 38. 33％,χ2 =3. 94, P<0. 05). The levels of HE4 and CAl25 were significantly lower than those in control group(HE4:(166. 5±45. 5) pmol/L vs. (245. 7±51. 8) pmol/L,t=6. 25,P<0. 05;CA125:(77. 4±18. 5) kU/L vs. (94. 4±16. 7) kU/L,t=3. 61,P<0. 05) . Conclusion Neoadjuvant chemotherapy combined with cytoreductive surgery can effectively improve the clinical efficiency and improve the prognosis of patients with advanced epithelial ovarian cancer.

Objective To compare the minimum 5?year follow?up outcomes of surgical management by posterior only, anterior only and combined posterior and anterior approaches for lumbar tuberculosis in adults, evaluate the mid?term follow?up results of the surgery for the treatment of lumbar tuberculosis and explore its advantages and indications. Methods From Jun 2004 to Jan 2010, 311 adult patients with lumbar tuberculosis were treated surgically. The clinical data of 137 cases that met the enrolled criteria and had integrity following?up data was analyzed retrospectively. It included the patients who had the surgical indication of the posterior only surgery but underwent the anterior only or the combined posterior and anterior ap?proaches before 2008. There were 83 cases of male and 54 cases of female. The age ranged from 20 to 75 years, with a mean of 65.6 years. Among these patients, 63 cases were treated with single?stage posterior debridement, interbody fusion and instru?mentation (the posterior group); 42 cases were treated with posterior instrumentation, and anterior debridement and bone graft in a single or two?stage procedures (the combined group) and 32 cases were treated with anterior debridement and strut graft?ing with instrumentation (the anterior group). Trauma index (the operation time, blood loss, the length of hospital stay, compli?cations);imaging parameters (Segment kyphotic angle, corrective rate, loss angle, bone fusion time) and the quality?of?life indi?cators (Oswestry Disability Index、Frankle grade、visual analogue scale、Macnab score) were compared among three groups. Re?sults The mean operation time, mean blood loss and the complications rate were (207.9 ± 30.9) min, (409.5 ± 107.9) ml and 12.95%in the posterior group;(270.7±32.0) min, (649.0±120.0) ml and 30.95%in the anterior group;(349.7±38.9) min, (840.0± 168.7) ml and 25%in the combined group. The operation time, blood loss and the complications rate of the posterior group were less than the anterior group and the combined group, and the difference was significant;The combined group consumed the longest operation time, associated with the most intraoperative blood loss, the highest complication rate and the longest hospital stay among the three groups, and the difference was significant. The correction rate of kyphosis achieved of the anterior group ( 52%± 5.45%) was significantly inferior to the posterior group (74%±5.04%) and the combined group (69%±7.95%), while the loss of cor?rection in the anterior group (2.5°) was higher than both the posterior group (0.8°) and the combined group (1.1°), and the differ?ence was significant. The average follow?up was(6.5±1.96)years (range, 5-11). The mean bone fusion time of the posterior group, the anterior group and the combined group were (6.0±1.5) months, (6.2±1.3) months and (6.5±1.6) months respectively, and there was no statistic difference. After the surgery, the quality of life was improved obviously in all patients. At the time of the latest fol?low?up, the improvement rate of the ODI,VAS and the excellent and good rate according to the Macnab score were 80.6%±2.1%, 81.7%± 1.6%and 95.24%in the posterior group;79.8%± 1.5%, 79.7%± 2.0%and 92.95%in the anterior group;81.3%± 1.1%, 79.9%±0.8%and 90.63%in the combined group. There was no significant difference among the groups in the improvement rates of the ODI, VAS, Frankel grade and the excellent and good rate of the Macnab score. Conclusion The Mid?term follow?up of the different surgical procedures for the treatment of the lumbar tuberculosis in adults were basically satisfactory. Compared with the traditional surgery, the posterior?only surgery is a safe, minimally invasive and effective method in the management of monoseg?ment lumbar tuberculosis in adults.

Objective To compare the minimum 5?year follow?up outcomes of surgical management by posterior only ap?proaches, anterior only approaches and combined posterior and anterior approaches for thoracic tuberculosis in adults, and evalu?ate the mid term follow?up results of posterior only approaches. Methods All of 184 patients with monosegment thoracic tubercu?losis between January 2003 and November 2010 were studied retrospectively. Among these patients, 62 cases were treated with posterior debridement combine with interbody fusion (PO group), 65 cases were treated by posterior instrumentation, anterior de?bridement and bone graft in one or two?stage procedures (AP Group ), and 57 cases were treated by anterior only approach (AO Group). The operation time, blood loss, Visual Analogue Scale, complications, recovery of neurological function, kyphosis angle, correction rate and loss angle were respectively compared between each group. Results Comparison of postoperative curative ef?fects showed:mean operation time and blood loss:PO group (260.05±30.75 min,735.95±161.43 ml) was better than AP group (411.65 ± 55.61 min, 1178.65 ± 184.50 ml)and AO group (343.65 ± 24.74 min, 965.35 ± 122.59 ml);corrective angle and correction rate:PO group (6.78°±1.13°, 72.48%±12.97%) and AP group (6.97°±1.05°, 73.10%±11.42%) were better than AO group (13.98°± 1.73°, 44.95%±16.84%);bed time:PO group and AO group were shorter than AP group. Mid term follow?up outcomes showed:ky?phosis angle and loss angle:PO group (8.56°±1.09°, 1.89°±1.41°) and AP group (8.55°±1.65°, 1.63°±1.11°) were better than AO group (16.39°±1.59°, 2.80°±1.29°);bone fusion time, VAS and recovery of neurological function:there were no statistically differ?ence in all groups. Conclusion The mid term follow?up outcomes of posterior debridement combined with interbody fusion is sat?isfied in the management of monosegment thoracic tuberculosis. It is a safe and effective method.

Objective: To determine the clinical efficacy and feasibility of multiple special formed titanium mesh cages (TMCs) to treat lumbo-sacral spinal tuberculosis via posterior approach. Methods: From July, 2007 to June, 2013, 25 patients with lumbo-sacral spinal tuberculosis underwent one-stage posterior debridement, internal if xation, and interbody fusion using multiple special formed titanium meshes. We compared the parameters as follow: the pre- and post-operative American Spinal Injury Association (ASIA) score, lumbo-sacral angle, the height of intervertebral space, visual analogue scale (VAS), and erythrocyte sedimentation rate (ESR), and observedoperation time, intraoperative blood loss, and time of bone gratf fusion. Results: Operation time ranged from 90 to 180 min, (128±24) min in average. Blood loss in the operation ranged from 100 to 800 mL, (310±125) mL in average. hTe patients were followed up for 24 to 59 months, (43±7) months in average. One patient delayed healing of wound. ASIA score was improved in a certain degree in patients with neurological dysfunctions. hTe lumbo-sacral angle and the height of intervertebral space in the post-operation were signiifcantly higher than those in the pre-operation (P<0.001). VAS was reduced obviously atfer 2 weeks of operation. hTe ESR recovered to the normal level 6 months atfer operation in all the patients. Solid fusion was achieved within 4 to 8 months, 6 months in average. No sinus tract, cerebrospinal meningitis, tuberculosis recurrence and titanium mesh subsidence were found. Conclusion: For lumbo-sacral tuberculosis, multiple special formed titanium mesh cages via posterior approach is safe and effective, which is good to the stability in spine reconstruction.

Objective To evaluate the clinical effect of one-stage posterior-anterior approach surgery for patients with cervical fracture and dislocation combined with locked facet.Methods A retrospective review was conducted on 21 cases of cervical dislocation and fracture combined with locked facet treated by one-stage posterior-anterior approach surgery between April 2011 and December 2012.There were 16 males and 5 females at age ranging from 23 to 61 years (mean,38.3 years).Posterior unlocking reduction by partial facetectomy and lateral mass screw fixation was performed,followed by anterior decompression,internal fixation and interbody fusion by titanium meshes.Outpatient or telephone follow-up was performed to evaluate bone fusion and recovery of neurologic function.Results Mean operation time was 140 minutes (130-210 minutes) and mean blood loss was 340 ml (range,150-600 ml).All incisions got primary healing with no operation-correlated complications.Five patients complicated with severe lung infection after surgery and one died of respiratory failure two week later.Titanium meshes achieved bone fusion within 3-9 months (mean,6 months) after surgery.At a mean follow-up of 17 months (range,12-30 months),there was no implant breakage and mesh displacement or collapse.According to the American Spinal Injury Association (ASIA) score,preoperative neurologic deficit restored by mean one grade at final follow-up.Conclusion One-stage posterior-anterior approach surgery is an ideal choice for cervical fracture and dislocation combined with locked facet,for it provides unlocking reduction,canal decompression,and rigid reconstruction of the anterior-posterior column.

Objective To investigate the complications of three-dimensional correction surgery for scoliosis and the corresponding prevention and treatment strategies.Methods From December 2004 to June 2011,727 cases of scoliosis were treated by three-dimensional correction system.There were 245 males and 482 females,aged from 3 to 62 years (average,18.2 years).Coronal Cobb angles ranged from 32° to 142° (average,87.6°),and sagittal Cobb angles ranged from-10° to 75° (average,45°).Results All patients were followed up for 12 to 90 months (average,62.5 months).All patients underwent surgery safely.The coronal correction rate ranged from 55％ to 98％ (average,85.2％),and the sagittal correction rate ranged from 35％ to 67％ (average,47.5％).There were no major complications such as death and nerve injury,but occurrences of other minor complications were 113 times in 102 cases.There were 26 cases of instrumentation-related complication,including 5 cases of pedicle screw loosening,5 cases of broken screw,8 cases of broken rod,3 cases of hook loosening,and 5 cases of pedicle fracture; 14 cases of them occurred early after correction surgery for adult scoliosis.There were 65 cases of correction-related complication,including 36 cases of junctional kyphosis (21 cases of proximal junctional kyphosis,11 cases of which were neuromuscular scoliosis; 15 cases of distal junctional kyphosis,including 4 cases of Marfan syndrome with scoliosis and 6 cases of neuromuscular scoliosis),22 cases of adding-on phenomenon,and 7 cases of flat back.Internal medicine complications included 6 cases of superior mesenteric artery syndrome and 7 cases of pulmonary complications.Operation-related complications included 4 cases of pressure sore and 5 cases of wound infection.Conclusion Accurate procedures of diagnosis and surgery for scoliosis are the key to decrease and prevent the complications.For adult scoliosis,enough instrumentation should be placed at the apical segments to decrease instrumentation-related complications.For neuromuscular scoliosis and Marfan syndrome with scoliosis,appropriate extension of fusion segment can effectively decrease junctional kyphosis.

BACKGROUND:Studies concerning how estrogen receptorβparticipates in bone metabolism are few now. OBJECTIVE:To investigate the effect of estrogen receptorβon the expression of osteoprotegerin and receptor activator of nuclear factor-κB ligand in human osteblast-like cells. METHODS:The retrovirus with the most effective interference sequence and non-specific short hairpin RNA was used to transfect human osteoblast-like cellMG63 in order to screen out the stable colon, and then amplified and cultured. The blank control and non-specific short hairpin RNA were used as control, and the stable inhibition rate of estrogen receptorβwas detected. The 17β-estradiol was added into the cells in three groups, that were MG63 cells, short hairpin RNA retrovirus estrogen receptorβ-mediated MG63 cells and negative control short hairpin RNA retrovirus-medicated MG63 cells, in order to detect the expressions of osteoprotegerin and receptor activator of nuclear factor-κB ligand mRNA in human osteoblast-like cells. RESULTS AND CONCLUSION: The human osteoblast-like MG63 cellline was further stably transfected with pRNAT-H1.4/Retro-estrogen receptorβshort hairpin RNA3, and then compared with the blank control and negative control, and found that estrogen receptorβcould express the stable inhibited human osteoblast-like cellline. The inhibition rate of estrogen receptorβmRNA was (88.17±1.17)%(P<0.05), and the inhibition rate of estrogen receptorβprotein was (89.01±1.22)%(P<0.05), indicating that estrogen receptorβgene knockdown human osteoblast-like cellmodels were constructed successful y. After estrogen intervention for 48 hours, the inhibition of MG63 cells with estrogen receptorβcould up-regulate the osteoprotegerin mRNA and protein expression in the blank control group and the negative control group (P<0.05), down-regulate the receptor activator of nuclear factor-κB ligand mRNA and protein expression (P<0.05), and up-regulate the osteoprotegerin receptor activator of nuclear factor-κB ligand expression. The results indicate that estrogen receptorβmay play an important role in bone metabolism through regulating osteoprotegerin/receptor activator of nuclear factor-κB ligand ratio.