As a heritable regulation , epigenetics can regulate gene expression by other ways without changing the DNA sequence ,and change cell or individual phenotypes .DNA methylation is an issue in the field of epigenetics research.Recently,many studies have been demonstrated that the methylation of repetitive DNA ,spe-cific gene and CpG island and loss of imprinting play an important role in tumor occurrence .As the development of technological approaches to DNA methylation ,we have a more comprehensive understanding on methylation pat-terns.As specific markers,abnormal methylation sites in the genome can be used in the diagnosis ,treatment and prognosis predictor of disease .For tumor development caused by DNA methylation ,the application of demethylat-ing drugs have achieved good effect in clinical treatment .

Objective:To investigate the efficacy and safety of perioperative aspirin use in elderly patients with osteoporotic vertebral fractures(OVF)undergoing vertebroplasty(VP).Methods:This was a retrospective cohort study.Clinical data of 136 OVF patients treated with VP in our department from Jan.2016 to Dec.2020 were analyzed.Differences in clinical data, treatment efficacy, intraoperative and postoperative complications and hematomas were compared between the aspirin group(n=71, receiving aspirin100 mg/d before VP and not taking other anticoagulant drugs)and the control group(n=65, not taking aspirin).Results:There was no significant difference in the analgesic score or physical activity scale score between the two groups before, 1 week after surgery and at the last follow-up( P>0.05). There were significant differences in the visual analog score(VAS), the analgesic score and the physical activity scale score before, 1 week after surgery and at the last follow-up within the aspirin group(7.12±1.33, 2.37±1.01 vs.2.63±1.04, 3.01±0.95, 1.56±0.65 vs.1.61±0.57, 2.75±0.53, 1.32±0.63 vs.1.44±0.52, P<0.01). No surgical site infection, injury of large vessels, intraspinal hematoma or pulmonary embolism was found in the aspirin group or the control group during the follow-up period.There was no difference in intraoperative or postoperative blood loss(12±3.5 ml vs.11.0±3.6ml, t=1.60), cement injection volume for a single vertebral body(4.5±1.9 ml vs.4.0±1.7 ml, t=1.40), cement spillage(14 cases or 19.7% vs.9 cases or 17.0%, χ2=0.15), nerve root irritation(3 cases or 4.2% vs.1 case 1.9%, χ2=0.43), re-fractures(5 cases 7.0 vs.3 cases 5.7%, χ2=0.10)or spinal epidural hematoma(8 cases or 11.3% vs.5 cases or 9.4%, χ2=0.11)between the aspirin group and the control group(all P>0.05). Conclusions:In OVF patients taking perioperative aspirin, the postoperative pain score, pain medication administration and mobility are significantly improved after VP, compared with pre-treatment.Surgical efficacy and safety show no significant difference between patients with and without aspirin administration.

Objective To evaluate efficacy,safety and complication of percutaneous kyphoplasty (PKP) for osteoporotic vertebral compression fracture (OVCF) in≥90 years patients.Methods Clinical data of 56 cases aged ≥ 90 years with osteoporotic vertebral compression fracture undergoing percutaneous kyphoplasty were retrospectively analyzed.Visual analog scale (VAS) score,analgesics administration score,locomotor activity score,bone cement leakage and incidence of refracture were evaluated before and 3 days after treatment,and at the last follow-up.Results The mean follow-up was 18.6 months (6-32 months) in all patients.The mean VAS score was (7.1 ±2.1) before treatment,(2.6±1.1) at 3 days after the procedure,and (1.8±0.7) at last follow-up,respectively (F=455.794,P＜0.001).Analgesics administration score were (2.0±1.7),(1.4±0.5) and (1.1±0.7) respectively before and 3 days after treatment,and at the last follow-up (F=9.631,P＜0.001).Locomotor activity score were (2.5±0.6),(1.2±0.5) and (1.0±0.3)before and 3 days after treatment,and at the last follow-up (F=63.254,P＜ 0.001) respectively.Bone cement leakage occurred in 10 cases(17.9％),recurrent fracture in 6 cases(10.7％),cerebrospinal leak in 3 cases (5.3％),and nerve root stimulation in2 cases(3.6％).Total complication rate was 33.9％(19/56),and all complications were transient and well tolerated.Conclusions Kyphoplasty for osteoporotic vertebral compression fracture in the very elderly is effective and safe.It alleviates fracture-induced pain,reduces analgesic drug use and improves spinal activity,and provides a better choice for minimal invasive treatment for nonagenarian OVCF patients.

Objective: To investigate the relationship between the melatonin and the development of testicle in rabbits. Methods: Thirty male rabbits (unreach puberty) were randomly divided into 2 groups:15 were actively immunized with melatonin and complete antigen, which was successfully synthesized through Mannich reaction by combining melatonin with BSA, the other 15 were taken as controls.The specific antibodies of rabbits were detected by ELISA and RIA.The serum testosterone(T) and luteinizing hormone(LH) were detected by RIA, the mass of testicle or epididymis and the number of sperm were measured. Results: The serum T and LH concentration in experiment group were significantly lower than that in the control group.The mass of testicle or epididymis and the number of sperm in experiment group were also significantly lower than that in the control group. Conclusion: The results suggest that the immunization of melatonin can block the development of testicle in male rabbits.

Objective:To investigate the impact of osteoporosis on clinical outcomes in elderly patients treated with short segment transforaminal lumbar interbody fusion.Methods:From May 2016 to May 2018, elderly patients who had undergone transforaminal lumbar interbody fusion(TLIF)in our hospital were retrospectively analyzed.Based on bone mineral density(BMD), patients were divided into the osteoporosis group(the OP group, n=75, T≤-2.5 in BMD)and the control group(the CO group, n=103, T>-1.0 in BMD). General patient information, clinical data and postoperative follow-up clinical results were compared between the two groups.Results:Eventually 178 cases were enrolled, including 68 with lumbar disc herniation and 110 with lumbar spinal stenosis.Preoperative Visual Analogue Scale(VAS)scores were 7.35±1.30 in the lower back and 7.32±1.30 in the leg for the OP group and 7.35±1.33 and 7.22±1.40, respectively, for the CO group, and there was no significant difference between the two groups( t=0.140 and 0.468, P=0.989 and 0.640). The proportions of cage collapse and internal fixation loosening were 70.7%(53/75)and 37.3%(28/75)in the OP group, which were higher than 22.3%(23/103)and 14.6%(15/103)in the CO group( χ2=41.440 and 12.280, both P=0.000). However, there was no significant difference between the OP group and the CO group in 1-and 2-year postoperative interbody fusion rates(postoperative 1-year rate: 89.3% or 67/75 vs.91.3% or 94/103, χ2=0.187, P=0.666; postoperative 2 year rate: 94.6% or 71/75 vs.95.1% or 98/103), χ2=0.021, P=0.885). There was no significant difference in VAS score and Oswestry disability index(ODI) between the OP group and the CO group at 6 months, 1 year and 2 years after surgery(all P>0.05). Conclusions:Although there are some osteoporosis-related complications such as cage subsidence and screw loosening, short-segment TLIF can still achieve good clinical results in elderly patients with osteoporosis.

Objective To investigate the effectiveness of open-door laminoplasty with lateral mass screw fusion in the treatment of multisegmental cervical spondylotic myelopathy (CSM)with segmental instability.Methods Forty-one patients with multisegmental cervical spondylotic myelopathy combined with instability underwent open-door laminoplasty with lateral mass screw fusion.The JOA(Japanese Orthopaedic Association)scoring system and Neck Disability Index(NDI) were applied to evaluate neurological function and axial neck/shoulder pain before and after surgery,respectively.In addition,the Ishihara method was used to measure the cervical curvature index(CCI),and spinal cord expansion was calculated with MRI.Results After 43 months of postoperative follow-up,there were significant differences in JOA scores[(7.12±0.91) vs.(14.73± 1.12),t =35.28,P＜0.001],areas of the dural sac at the level of maximum stenosis in MRI[(0.22±0.05)]cm2 vs.(0.62±0.09)cm2,t=14.15,P＜0.001],and NDI scores[(24.46±6.61) vs.(12.90±3.46),t=15.59,P ＜ 0.001],compared with those before treatment.However,there were no significant differences in pre-and post-operation CCI[(10.36±1.69) vs.(10.87±2.05),t=1.11,P＞0.05].Conclusions Open-door laminoplasty with lateral mass screw fusion can improve neurological function and maintain the normal cervical curvature.Therefore,it is effective in the treatment of multisegmental cervical spondylotic myelopathy with segmental instability.

Objective:To examine the correlation between intraoperative pain scores during puncturing and postoperative complications in elderly patients with osteoporotic vertebral fractures(OVF)treated with vertebroplasty(VP).Methods:In a retrospective case-control study, clinical data of 326 patients with single-segment OVF treated with VP, including 42 patients(12.9%)(the complication group)with complications within 1 month of surgery and 284 patients(87.1%)without complications(the control group), were compared.Changes in patient numerical evaluation scale(NRS)scores were recorded and compared for the complication group and the control group at different time points, which concluded preoperative(T0), intraoperative puncturing of soft tissues(T1), bone puncturing(T2), bone cement injection into the vertebral body(T3), 24 hours(T4), 1 month(T5)and 3 months(T6)after surgery.Results:NRS scores for patients in the complication group vs.those in the control group at different phase were(5.78±2.11 vs.6.10±2.21)points at T0, (7.59±1.46 vs.4.63±0.86)points at T1, (7.30±1.35 vs.5.14±1.07)points at T2, (6.97±1.24 vs.6.11±1.58)points at T3, (4.09±0.82 vs.2.19±0.87)points at T4, (2.07±0.80 vs.1.93±0.78)points at T5, and(1.83±0.72 vs.1.74±0.65)points at T6, but there was no significant difference between the two groups at T0( P>0.05).The complication group had higher NRS scores than the control group at T1, T2, T3, and T4(all P<0.05).For intra-group comparisons, both the complication group and the control group showed statistically significant differences between T0 and T4, between T0 and T5, and between T4 and T5(all P<0.05). Conclusions:Elderly OVF patients who are treated with VP and exhibit post-surgery complications often experienced severe pain during surgery, and an NRS score greater than 7 may be an independent risk factor for postoperative complications of VP.Effort should be made to avoid or reduce complications related to surgery, reduce pain and improve treatment outcomes of VP for elderly patients.

Objective:To compare the minimally invasive transforaminal lumbar interbody fusion (MTLIF) with open transforaminal lumbar interbody fusion (OTLIF) in treatment of lumbar degenerative disease.Methods:Clinical data of 63 patients with single segment lumbar degenerative disease treated in Department of Orthopedics of Beijing Hospital from November 2015 to September 2016 were retrospectively analyzed, among whom 30 cases received MTLIF and 33 cases received OTLIF. The operative time, intraoperative X-ray exposure times, intraoperative blood lose, postoperative drainage,perioperative fever, adjacent segment degeneration, loosening of internal fixation and cage collapse were observed in two groups 4 years after operation, and the visual analog scale (VAS) score of the lower back and the leg, the Oswestry disability index (ODI) score were compared between two groups.Results:The operation time [(191.6±50.5) min] and radiation exposure times [(15.5±6.4) times] in MTLIF group were significantly more than those in OTLIF group [(105.8±23.1) min, (7.2±1.4)times, t=17.210, t=10.850,all P<0.01]. The intraoperative blood loss [(150.4±70.4) ml], postoperative drainage [(90.4±30.7)ml], VAS score (2.4±0.7) and ODI score (24.5±3.7) 2 weeks after surgery in MTLIF group were significantly lower than those in OTLIF group [(250.7±43.9)ml,(216.3±67.8)ml,(4.5±1.6),(30.6±4.6), t=-12.830, t=-14.070, t=-6.890, t=-5.805,all P<0.01]. There were no significant differences in the incidence of fever [1 case(3.3%) vs. 4 cases(12.1%),χ2=-1.661, P=0.20], VAS score[(1.2±0.7) vs. (1.3±0.6), t=-0.628, P=0.53], ODI score[(14.2±2.7) vs. (14.7±2.5), t=-0.756, P=0.45], fusion rate of Bridwell grade Ⅰ [86.7%(26/30) vs. 84.8%(28/33),χ2=0.042, P=0.84] 1 year after surgery; and the adjacent segment degeneration [0 case(0) vs. 1 case(3.0%),χ2=0.924, P=0.34], internal fixation loosening [1 case(3.3%) vs. 1 case(3.0%),χ2=0.005, P= 0.95] and cage collapse 4 years after surgery [1 case(3.3%) vs. 1case(3.0%),χ2=0.005, P=0.95] between MTLIF group and OTLIF group. Conclusion:Compared with OTLIF, MTLIF has longer operation time and more radiation exposure, but it can achieve full decompression, the same fusion rate, less bleeding, less trauma, faster recovery, fewer complications and satisfactory long-term effect.

ObjectiveTo investigate the mechanism of Atractylodis Macrocephalae Rhizoma（AMR） in the treatment of slow-transmission constipation（STC） by observing the effects of AMR on short-chain fatty acids and intestinal barries in STC mice. MethodForty-eight male KM mice were randomly divided into blank group， model group， AMR low-， medium-， high-dose groups（2.5， 5， 10 g·kg^-1） and mosapride group（2.5 mg·kg^-1）. Except for the blank group， all groups were gavaged with loperamide suspension（5 mg·kg^-1） twice daily for 14 d to construct the STC mouse model. At the same time， each drug administration group was given the corresponding drug by gavage for consecutive 14 d， the blank and model groups were gavaged with equal volume of distilled water. The effects of the treatment of AMR on body mass， defecation frequency， fecal water content and intestinal propulsion rate of mice were observed， the pathological changes of mouse colon were observed by hematoxylin-eosin（HE） staining and periodic acid-Schiff（PAS） staining， the levels of gastrin（GAS） and motilin（MTL） in serum were detected by enzyme-linked immunosorbent assay（ELISA）， gas chromatography-mass spectrometry（GC-MS） was used to detect the contents of short-chain fatty acids（SCFAs） in mouse feces， real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） and Western blot were used to determine the mRNA and protein expression levels of zonula occludens-1（ZO-1）， Occludin， and Claudin-1 in the colon of mice. ResultCompared with the blank group， the body mass， defecation frequency， fecal water content and intestinal propulsion rate of mice in the model group were significantly decreased（P<0.05， P<0.01）， the arrangement of colonic tissues was disordered， and the number of goblet cells was reduced， the levels of GAS and MTL in serum were significantly decreased（P<0.01）， and the levels of SCFAs in the feces were on a decreasing trend， with the contents of acetic acid， propionic acid， butyric acid， isobutyric acid and valeric acid were significantly decreased（P<0.05， P<0.01）， the mRNA and protein expression levels of ZO-1， Occludin and Claudin-1 in the colonic tissues were significantly decreased（P<0.01）. The above results suggested that STC mouse model was successfully constructed. Compared with the model group， the body mass， defecation frequency， fecal water content and intestinal propulsion rate of mice in AMP administration groups all increased significantly（P<0.05， P<0.01）， the mucosal layer of the colonic tissues was structurally intact without obvious damage， and the number of goblet cells increased， serum levels of GAS and MTL were significantly increased（P<0.01）， the contents of SCFAs in the feces were all on a rising trend， with the contents of acetic， propionic， butyric and isobutyric acids rising significantly（P<0.05， P<0.01）， the mRNA and protein expression levels of ZO-1， Occludin and Claudin-1 in the colonic tissues were significantly increased（P<0.05， P<0.01）. ConclusionAMR is able to improve the constipation symptoms in STC mice， and its mechanism may be related to increasing the contents of SCFAs in the intestine as well as promoting the mRNA and protein expression levels of ZO-1， Occludin and Claudin-1 in the colon.

Whether direct manipulation of Parkinson’s disease (PD) risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue. Here, we used an adeno-associated virus serotype 9 (AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras (SNs) of two monkey groups: an old group and a middle-aged group. After the operation, the old group exhibited all the classic PD symptoms, including bradykinesia, tremor, and postural instability, accompanied by key pathological hallmarks of PD, such as severe nigral dopaminergic neuron loss (>64%) and evident α-synuclein pathology in the gene-edited SN. In contrast, the phenotype of their middle-aged counterparts, which also showed clear PD symptoms and pathological hallmarks, were less severe. In addition to the higher final total PD scores and more severe pathological changes, the old group were also more susceptible to gene editing by showing a faster process of PD progression. These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys. Taken together, this system can effectively develop a large number of genetically-edited PD monkeys in a short time (6–10 months), and thus provides a practical transgenic monkey model for future PD studies.