Objective:To analyze the methylation characteristics of the lymphocyte-specific protein-tyrosine kinase (LCK) promoter region in the peripheral blood circulation of rheumatoid arthritis (RA) patients and its correlation with clinical indicators.Methods:Targeted methylation sequencing was used to compare the methylation levels of 7 CpG sites in the LCK promoter region in the peripheral blood of RA patients with healthy controls (HC) and osteoarthritis (OA) patients. Correlation analysis and ROC curve construction were performed with clinical information.Results:Non-parametric tests revealed that compared with HC [0.53(0.50, 0.57)] and OA patients [0.59(0.54, 0.62), H=47.17, P<0.001], RA patients [0.63(0.59, 0.68)] exhibited an overall increase in methylation levels. Simultaneously, when compared with the HC group [0.38(0.35, 0.41), 0.59(0.55, 0.63), 0.60(0.55, 0.64), 0.59(0.55, 0.63), 0.58(0.53, 0.62), 0.45(0.43, 0.49), 0.57(0.54, 0.61)], the RA group [0.46(0.42, 0.49), 0.70(0.65, 0.75), 0.70(0.66, 0.76), 0.70(0.65, 0.75), 0.69(0.64, 0.74), 0.55(0.51, 0.59), 0.68(0.63, 0.73)] showed a significant elevation in methylation levels at CpG sites cg05350315_60, cg05350315_80, cg05350315_95, cg05350315_101, cg05350315_104, cg05350315_128, and cg05350315_142, with statistically significant differences ( Z=-5.63, -5.89, -5.91, -5.89, -5.98, -5.95, -5.95, all P<0.001). Compared with the OA group [0.65(0.59, 0.69), 0.65(0.60, 0.69), 0.64(0.58, 0.68), 0.50(0.45, 0.54), 0.63(0.58, 0.67)], the RA group [0.70(0.66, 0.76), 0.70(0.65, 0.75), 0.69(0.64, 0.74), 0.55(0.51, 0.59), 0.68(0.63, 0.73)] exhibited a significant increase in methylation levels at CpG sites cg05350315_95, cg05350315_101, cg05350315_104, cg05350315_128, and cg05350315_142, with statistically significant differences ( Z=-3.56, -3.52, -3.60, -3.67, -3.62; P=0.036, 0.042, 0.031, 0.030, 0.030). Furthermore, Pearson correlation coefficient analysis revealed a positive correlation between the overall methylation level in this region and C-reactive protein (CRP) ( r=0.19, P=0.004) and erythrocyte sedimentation rate ( r=0.14, P=0.035). The overall methylation level of the LCK promoter region in the CRP (low) group [0.63 (0.58, 0.68)] was higher than that in the CRP (high) group [0.65(0.61, 0.70)], with statistically significant differences ( Z=2.60, P=0.009). Finally, by constru-cting a ROC curve, the discriminatory efficacy of peripheral blood LCK promoter region methylation levels for identifying RA patients, especially seronegative RA patients, from HC and OA groups was validated, with an AUC value of 0.78 (95% CI: 0.63, 0.93). Conclusion:This study provides insights into the methylation status and methylation haplotype patterns of the LCK promoter region in the peripheral blood of RA patients. The overall methylation level in this region is positively correlated with the level of inflammation and can be used to differentiate seronegative RA patients from the HC and OA patients.

[Objective]To investigate primary and secondary transfer of exfoliated cells from human hands on non-porous substrates such as plastic steering wheel or computer mouse.[Methods]DNA detection sensitivity and detection limit for mixed DNA profiling were examined to understand our laboratory's ability to test for trace DNA.Forensic swabs were used to collect samples from volunteers'one-hour-long unwashed hands,substrates touched by volunteers'immediately or 30 min following shaking hands,and individual A's daily-use substrates touched by individual B and then by individual A again.Simulations were conducted to assess the potential for introduction of another person's exfoliated cells from hands into routine casework samples.[Results]Our laboratory can obtain a full DNA profile from as little as 0.020 ng of DNA and detect minor components in a 1:9 mixed DNA sample.85%of samples from unwashed hands yielded a full DNA profile.Primary transfer of a full DNA profile was found in 77%of substrates touched by volunteers'dominant hand 30 min after hand washing,allowing differentiation between good and poor shedders,with no significant difference in genders and substrate types.75%of substrates touched 30 min after hand washing and then immediately following handshaking yielded the other individual's DNA profile(secondary transfer),with the number of short tandem repeat(STR)loci detected ranging from 0 to 23;the percentage and number decreased substantially when the substrates were touched 30 minutes later.No foreign DNA was detected in routine casework samples with introduced exfoliated cells from hands.When two individuals took turns touching items with their hands,the major contributor to the DNA profile was not always the individual who made the last contact.[Conclusions]Primary and secondary DNA transfer can be detected on non-porous substrates,and based on the deposit of hand exfoliated cells,individuals can be categorized as good or poor shedders,which is an important factor affecting detection of DNA transfer.Besides considering the laboratory's DNA detection sensitivity,if DNA is detected on substrates by hand contact,we need to take into account the potential for secondary transfer at different levels of activity when interpreting the results.

BACKGROUND: The derivation of salivary gland (SG) progenitors from pluripotent stem cells (PSCs) presents significant potential for developmental biology and regenerative medicine. However, the existing protocols for inducing SG include limited factors, making it challenging to mimic the in vivo microenvironment of embryonic SGs. METHODS: We reported a cocktail factor approach to promote the differentiation of mouse embryonic stem cell (mESC)-derived oral epithelium (OE) into SG progenitors through a three-dimensional co-culture method. Upon confirming that the embryonic SG can promote the differentiation of mESC-derived OE, we performed RNA sequence analysis to identify factors involved in the differentiation of SG progenitors. RESULTS: Our findings highlight several efficient pathways related to SG development, with frequent appearances of four factors: IFN-c, TGF-b2, EGF, and IGF-1. The combined treatment using these cocktail factors increased the expression of key SG progenitor markers, including Sox9, Sox10, Krt5, and Krt14. However, absence of any one of these cocktail factors did not facilitate differentiation. Notably, aggregates treated with the cocktail factor formed SG epitheliallike structures and pre-bud-like structures on the surface. CONCLUSION In conclusion, this study offers a novel approach to developing a differentiation protocol that closely mimics the in vivo microenvironment of embryonic SGs. This provides a foundation for generating PSC-derived organoids with near-physiological cell behaviors and structures.

Objective To evaluate the clinical efficacy and safety of CT-guided percutaneous osteoplasty(POP)in the treatment of osteolytic metastases of the pelvis.Methods The clinical data of a total of 40 patients with pelvic osteolytic metastases,who received CT-guided POP at the Affiliated Zhongda Hospital of Southeast University between October 2011 and December 2021,were collected.Visual analogue scale(VAS)score was used to evaluate the clinical pain relief degree at one week,one month,3 months,6 months and 12 months after POP,and the joint function and the used dose of analgesic drugs were recorded.The preoperative and the postoperative 3-month,6-month and 12-month extents of the pelvic tumor destruction were compared.Based on the progression of local lesions within 12 months of follow-up,the patients were divided into controlled group and progression group.The proportion of using systemic anti-tumor therapy,the size of lesion,the amount of bone cement injected,and the cement filling ratio were compared between the two groups.Results Successful surgical procedure was accomplished for 57 lesions in 40 patients.The mean amount of bone cement injected was(4.56±2.25)mUpoint.In the 40 patients,the preoperative and the postoperative one-week,one-month and 3-month VAS score were(8.00±0.85)points,(2.05±0.96)points,(2.08±0.94)points and(2.18±0.84)points respectively,the difference in VAS score between preoperative value and postoperative one-week value was statistically significant(P＜0.01).In 37 patients,the postoperative 6-month VAS score was(2.35±0.54)points;and in 28 patients,the postoperative 12-month VAS score was(2.43±0.79)points.The differences in VAS score between postoperative one-week value and postoperative one-month,3-month,6-month,and 12-month values were not statistically significant(all P＞0.05),while the differences in VAS score between preoperative value and postoperative values were statistically significant(F=316.3,P＜0.01).The postoperative 3-month,6-month,and 12-month local control rates were 96.49％,85.19％,and 78.12％respectively,the differences between each other among the above three values were statistically significant(P=0.026).No statistically significant differences in the proportion of using systemic anti-tumor therapy,the lesion size and the amount of bone cement injected existed between the controlled group and the progression group(all P＞0.05).The cement filling ratio in the controlled group and the progression group was(81.26±9.17)％and(68.40±12.98)％respectively,and the difference between the two groups was statistically significant(P＜0.01).Conclusion For the treatment of pelvic metastases,CT-guided POP is clinically safe and effective.The injected bone cement can control the progression of local lesions for a longer time.(J Intervent Radiol,2023,32:1197-1201)

Objective:To analyze the incidence and epidemiological characteristics of traumatic spinal cord injury in China in 2018.Methods:Multi-stage stratified cluster sampling was used to randomly select hospitals capable of treating patients with spinal cord injury from 3 regions，9 provinces and 27 cities in China to retrospectively investigate eligible patients with traumatic spinal cord injury admitted in 2018. National and regional incidence rates were calculated. The data of cause of injury，injury level，severity of injury，segment and type of fracture，complications，death and other data were collected by medical record questionnaire，and analyzed according to geographical region，age and gender.Results:Medical records of 4，134 patients were included in this study，with a male-to-female ratio of 2.99∶1. The incidence of traumatic spinal cord injury in China in 2018 was 50.484 / 1 million （95% CI 50.122-50.846）. The highest incidence in the Eastern region was 53.791 / 1 million （95% CI 53.217-54.365）. In the whole country，the main causes of injury were high falls （29.58%），as well as in the Western region （40.68%），while the main causes of injury in the Eastern and Central regions were traffic injuries （31.22%，30.10%）. The main injury level was cervical spinal cord in the whole country （64.49%），and the proportion of cervical spinal cord injury in the Central region was the highest （74.68%），and the proportion of lumbosacral spinal cord injury in the Western region was the highest （32.30%）. The highest proportion of degree of injury was incomplete quadriplegia （55.20%），and the distribution pattern was the same in each region. A total of 65.87% of the patients were complicated with fracture or dislocation，77.95% in the Western region and only 54.77% in the Central region. In the whole country，the head was the main combined injury （37.87%），as well as in the Eastern and Central regions，while the proportion of chest combined injury in the Western region was the highest （38.57%）. A total of 32.90% of the patients were complicated with respiratory complications. There were 23 patients （0.56%） died in hospital，of which 17（73.91%） died of respiratory dysfunction. Conclusions:The Eastern region of China has a high incidence of traumatic spinal cord injury. Other epidemiological features include high fall as the main cause of injury cervical spinal cord injury as the main injury level，incomplete quadriplegia as the main degree of injury，head as the main combined injury，and respiratory complications as the main complication.

Objective To assess the quantitative impact of the application of high-throughput sequencing worldwide on clinical virology.Methods The following records from 2000 to 2018 were collected from National Center for Biotechnology Information:The number of complete genome of virus records per year;the number of virus discovery articles per year;the number of virus quasi-species articles per year;the number of viral infection gene signature articles per year.The method of interrupted time series was applied to analyze the trends of those records.Results Since the sequencing centers worldwide began to transit from the Sanger sequencing to the high-throughput sequencing technology in 2008,the annual value of records mentioned above increased by 3.755,2.760,6.195 and 3.885 times respectively.The long-term trends of change of each record are as follows:the number of complete genome of virus records increased by 1639.991 per year (P＜0.001);the number of virus discovery articles increased by 83.091 per year (P＜0.001);the number of virus quasi-species articles increased by 2.509 per year (P＜ 0.001);the number of viral infection gene signature articles increased by 30.836 per year (P＜ 0.001).Conclusions In 2008,the sequencing centers worldwide began to apply high-throughput sequencing technology which led to continuous decrease of the cost for sequencing.It result ed in not only increasingly enriched applications in clinical virology but also a long-term positive impact on the advance of clinical virology.

Objective To investigate the feasibility and primary clinical outcomes of a kind of independent research and development customized 3D printed external fixator in the treatment of tibial fracture.Methods Data of 7 tibial fracture patients who were treated using computer-assisted reduction and 3D printing technique including a new kind of customized external fixator with automatical reduction function from December 2013 to November 2015 were retrospectively analyzed.There were 5 males and 2 females in this cohort.The age of the patients ranged from 25 to 49 years old,with an average age of 38.1 years old.The procedure was as follows.At first,two or three pins were placed separately into the distal and proximal tibia away from fracture site.Then,the CT scanning was performed on the fractured tibia.According to the CT scanning data,the external fixator was designed by computer software and manufactured by the 3D printing technique.The reduction of tibial fracture was achieved automatically after the 3D printed external fixator was connected to the inserted pins.Finally,the accuracy of reduction was assessed by postoperative X-ray image.During the different fracture healing course,the stress environment could be dynamically adjusted by screwing or unscrewing the nuts to fulfill rigid fixation,compression fixation or elastic fixation for the bone healing.Results All the 7 cases had obtained successful reduction by only one time operation with the 3D printed external fixators.The average lateral angulation was 1.42°± 1.13°,and the average anteroposterior angulation was 1.65°± 1.36°,while the average lateral displacement was 1.38± 1.44 mm,and the average anteroposterior displacement was 1.83± 1.30 mm,which were measured from postoperative X-ray image.The individual durations of the operation were 9 min,8 min,9 min,1.5 min,1.4 min,1.4 min and 2.4 min,respectively.All fractures had achieved bony union in 19-25 weeks after operation,with an average time of 21.4±1.6 weeks.All patients were followed up for 6 months when removal of the external fixator,and no refracture occurred.Conclusion The customized 3D printed external fixator has the function of automatical reduction.In addition,it can provide three kinds of fixation modes,which are rigid fixation,compression fixation and elastic fixation.It has the advantages of simple operation,accurate reduction,reasonable fixation,etc.

BACKGROUND: Bone tissue engineering based on pluripotent stem cells (PSCs) is a new approach to deal with bone defects. Protocols have been developed to generate osteoblasts from PSCs. However, the low efficiency of this process is still an important issue that needs to be resolved. Many studies have aimed to improve efficiency, but developing accurate methods to determine efficacy is also critical. Studies using pluripotency to estimate efficacy are rare. Telomerase is highly associated with pluripotency. METHODS: We have described a quantitative method to measure telomerase activity, telomeric repeat elongation assay based on quartz crystal microbalance (QCM). To investigate whether this method could be used to determine the efficiency of in vitro osteogenic differentiation based on pluripotency, we measured the pluripotency pattern of cultures through stemness gene expression, proliferation ability and telomerase activity, measured by QCM. RESULTS: We showed that the pluripotency pattern determined by QCM was similar to the patterns of proliferation ability and gene expression, which showed a slight upregulation at the late stages, within the context of the general downregulation tendency during differentiation. Additionally, a comprehensive gene expression pattern covering nearly every stage of differentiation was identified. CONCLUSION: Therefore, this assay may be powerful tools for determining the efficiency of differentiation systems based on pluripotency. In this study, we not only introduce a new method for determining efficiency based on pluripotency, but also provide more information about the characteristics of osteogenic differentiation which help facilitate future development of more efficient protocols.
Bone and Bones ; Down-Regulation ; Gene Expression ; In Vitro Techniques ; Methods ; Mouse Embryonic Stem Cells ; Osteoblasts ; Pluripotent Stem Cells* ; Quartz Crystal Microbalance Techniques ; Telomerase* ; Up-Regulation

Objective To investigate whether initial intervention based on WBC measured within 2 hours postoperatively can reverse the uroseptic shock induced by UUTEL.Methods From May,2015 to July 2015,24 female New Zealand rabbits,weighing 2.0-2.5 kg,born 2-3 months,used as uroseptic shock model.Their ureters were ligated and followed by injection of Escherichia coli solution into the renal pelvis.Then,those animals were randomly assigned into control group(the first group)and experimental groups (the second group,the third group).Each group had 8 rabbits initially.The first group did not receive sensitive antibiotic or fluid resuscitation.The second group received imipenem and cilastatin sodium 15mg/kg and normal saline 5 ml/kg 2 hours postoperatively.The third group received sensitive antibiotic and fluid resuscitation the same dosage as the second group 6 hours postoperatively.Mean arterial pressure(MAP) was recorded for 10 hours and survival rate of all groups for 72h postoperatively was recorded..The clinical data of 46 patients whose WBC count less than 2.85 × 109/L within two hours after UUTEL were analyzed retrospectively.These patients were divided into two groups based on the time of intervention.Group A including 19 patients received routine antibiotic,fluid therapy,low-dose corticosteroids when there was symptom of shock.Group B including 27 patients immediately received resuscitation bundle protocol when there was a drastic decrease in WBC.The incidence of septic shock,the rate of intubation,length of stay in ICU,length of stay in hospital postoperatively,hospitalization cost and survival rate of these two groups were compared.Results All 8 rabbits of the first group died within 72 h,postoperatively and the median time of survival was 11 h.None of the second group rabbits developed shock and all rabbits survived 72 h after operation.6/8 rabbits of the third group survived 72 h after operation.15 patients in Group A and 3 patients in Group B experienced acute uroseptic shock (P ＜ 0.05).11 patients in Group A and one patients in Group B underwent tracheal intubation (P ＜ 0.05).18 patients in Group A and 6 patients in Group B were transferred to ICU(P ＜ 0.05),and their length of stay in ICU was (10.8 ± 5.4) d and (7.5 ± 2.8) d,respectively(P ＞ 0.05).The length of stay in hospital and hospitalization cost of Group A and B were (19 ± 9.8)d vs.(7 ±4.7)d(P ＜0.05),(94 583 ±51 623) RMB vs.(35 389 ± 16 342) RMB respectively (P ＜ 0.05).One patient in Group A died due to acute uroseptic shock and none of Group B died.Conclusions Our animal model and clinical cohort study showed that initial intervention based on WBC mneasured within 2 hours postoperatively can reverse the uroseptic shock induced by UUTEL and improve the prognosis.

Objective To preliminary evaluate the mid-term clinical effect of endovascular repair in treating spontaneous extracranial internal carotid artery (ICA) dissection,and to observe the patency of stent.Methods The clinical data and imaging materials of 6 patients with spontaneous extracranial ICA dissection,who were treated with endovascular repair during the period from March 2012 to December 2012,were retrospectively analyzed.The U.S.National Institute of Heahh Stroke Scale (NIHSS) scores were determined before and after endovascular repair,and the postoperative stent patency condition was assessed,the results were analyzed.Results A total of 6 patients,including 4 males and 2 females with a median age of 50 years old (40.75-54.75 years old),received endovascular repair therapy.The median interval from the onset of disease to accept endovascular treatment was 10 days (one week-3 months).After the implantation of stent,the blood flow in the true lumen returned to normal immediately,although part of the false lumen was still filled with contrast agent.Embolism of retinal artery occurred in one patient during the operation,no death occurred.The median follow-up time was 54.4 months (49.7-57.9 months).The NIHSS score determined at the last follow-up visit was not significantly different from the preoperative one (P=0.102).Imaging reexamination revealed that the false lumen at the ICA stent segment disappeared in all 6 patients,and no obvious in-stent stenotic changes were observed.Conclusion Endovascular therapy of selected symptomatic extracranial carotid artery dissection with bare stents can effectively prevent the recurrence of clinical symptoms and promote ICA remodeling with excellent mid-term patency.