1.Analysis of recurrent cervical abscess: a case report.
Junxin WANG ; Hui WANG ; Maogang SUN ; Endong ZHANG ; Zhonghua ZHANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2025;39(6):574-577
This report presents a case of esophageal fistula with recurrent cervical abscess as the initial symptom. The patient underwent anterior cervical fracture reduction and internal fixation in 2021 due to trauma. In 2023, the patient received radiofrequency ablation for bilateral thyroid lobe lesions, after which recurrent cervical abscesses occurred over an 8-month period. Fiberoptic laryngoscopy with the biopsy channel insufflation method revealed a large esophageal fistula at the esophageal inlet. After removing the internal fixation plate, the patient received daily wound care, and serial fiberoptic laryngoscopy revealed spontaneous healing of the esophageal fistula. During the 1-year follow-up, no recurrence of cervical abscess was observed, and the esophageal fistula healed well.
Humans
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Abscess
;
Recurrence
;
Neck
;
Female
;
Adult
;
Fracture Fixation, Internal
2.Comparative study of different treatment methods for peroneus longus tendon stump in anterior cruciate ligament reconstruction.
Peng WANG ; Zhiwei LIU ; Zhonghua SHI ; Fan ZHAO ; Jiawen WANG ; Huan LI
Chinese Journal of Reparative and Reconstructive Surgery 2025;39(12):1526-1531
OBJECTIVE:
To compare the impact of different peroneus longus tendon (PLT) stump management techniques on ankle function following arthroscopic anterior cruciate ligament (ACL) reconstruction with autologous PLT grafts.
METHODS:
A retrospective analysis was conducted on 60 patients with ACL rupture who met the inclusion criteria between August 2020 and July 2024. All patients underwent arthroscopic ACL reconstruction using the autologous PLT grafts. Patients were assigned to group A [PLT stump sutured to peroneus brevis tendon (PBT), n=30] or group B (no stump intervention, n=30). The two groups showed no significant difference ( P>0.05) in baseline data, including gender, age, body mass index, injury mechanism, affected side, preoperative American Orthopaedic Foot and Ankle Society (AOFAS) score, ankle muscle strength, ankle range of motion, and arch-related angles. Postoperative outcomes were assessed using the AOFAS score, ankle muscle strength (eversion and first-ray plantar flexion), arch-related angles (medial/lateral longitudinal and anterior/posterior arch), ankle range of motion (eversion, inversion, dorsiflexion, plantarflexion), and limb symmetry index (LSI). Change values in muscle strength, arch-related angles, and range of motion from preoperative to 12 months postoperatively were calculated for intergroup comparison.
RESULTS:
Groups A and B had comparable PLT graft lengths and diameters ( P>0.05). All patients were followed up 13-16 months (mean, 14.5 months). Postoperative complications included 1 case of incision infection, 1 case of deep vein thrombosis, and 1 case of knee stiffness in group A, 1 case of knee stiffness in group B. There was no significant difference in the overall complication incidences between groups ( P>0.05). No significant difference was found in the AOFAS scores between different time points and between groups ( P>0.05). At 12 months after operation, neither group showed significant changes from preoperative baseline in ankle strength, range of motion, or arch-related angles, and there was no significant difference in these change values between groups ( P>0.05). There was no significant difference in LSI between the two groups at 6 or 12 months postoperatively ( P>0.05).
CONCLUSION
Both suturing and leaving the PLT stump untreated during arthroscopic ACL reconstruction provided comparable ankle outcomes and well-preserved foot and ankle function.
Humans
;
Anterior Cruciate Ligament Reconstruction/methods*
;
Male
;
Retrospective Studies
;
Female
;
Adult
;
Anterior Cruciate Ligament Injuries/surgery*
;
Tendons/transplantation*
;
Range of Motion, Articular
;
Arthroscopy/methods*
;
Young Adult
;
Treatment Outcome
;
Muscle Strength
;
Transplantation, Autologous
;
Ankle Joint/surgery*
;
Middle Aged
;
Adolescent
3.Current situation investigation and analysis of influencing factors on the long-term quality of life of cured and discharged patients with severe acute pancreatitis.
Wenjun ZHOU ; Pinjie ZHANG ; Weili YU ; Zhonghua LU ; Mingjuan LI ; Lijun CAO ; Lu FU ; Shaokang WANG ; Yun SUN
Chinese Critical Care Medicine 2025;37(2):146-152
OBJECTIVE:
To investigate the current status of long-term quality of life in patients with severe acute pancreatitis (SAP) who have been cured and discharged, and to analyze the influencing factors affecting long-term quality of life in SAP cured patients after discharge.
METHODS:
A retrospective collection was conducted. Patients who were received standardized treatment before being cured and discharged from the hospital admitted to the first department of critical care medcine of the Second Affiliated Hospital of Anhui Medical University from January 2017 to December 2023 were enrolled. According to the 36-item short form health survey scale (SF-36) score, patients were divided into high score group (high quality of life, the top 50% of patients with total SF-36 score) and low score group (low quality of life, the bottom 50% of patients with total SF-36 score). The gender, age, history of hypertension and diabetes, etiology of pancreatitis, acute physiology and chronic health evaluation II (APACHE II), sequential organ failure assessment (SOFA), CT severity index (CTSI), laboratory indicators such as C-reactive protein (CRP), procalcitonin (PCT), blood glucose, and triglycerides upon admission, use of vasoactive drugs, non-invasive/high-flow ventilation, invasive ventilation, retroperitoneal puncture and drainage, open pancreatic surgery treatment and secondary infection during hospitalization were collected, as well as the retention of abdominal drainage tubes at discharge from hospital. Distribute follow-up questionnaires or telephone follow-up surveys through WeChat and Question Star programs to investigate the pancreatic secretion function, chronic abdominal pain, and recurrence of pancreatitis of patients after discharge. Multivariable Logistic regression was used to analyze the relevant factors affecting the long-term quality of life of cured patients with SAP.
RESULTS:
A total of 86 patients were ultimately enrolled. There were 43 patients in both the high and low score groups. Among 86 patients, 20 experienced acute pancreatitis recurrence, with a recurrence rate of 23.26%. Twenty-two (25.58%) experienced chronic abdominal pain after discharge, and 5 patients (5.81%) needed medication to relieve pain. Thirty-three patients (38.37%) had pancreatic exocrine dysfunction after discharge, characterized by abdominal distension, constipation or diarrhea. Twenty-two patients (25.58%) suffered from pancreatic endocrine dysfunction, and were diagnosed with diabetes. Univariate analysis showed that compared with the high score group, the low score group had more patients with hypertension, initial renal dysfunction, initial severe metabolic acidosis, initial serum calcium < 2.0 mmol/L, blood glucose > 11.1 mmol/L and cultured Gram positive bacteria (from blood/body fluid/pancreatic necrotic tissue) during treatment (48.84% vs. 16.28%, 60.47% vs. 32.56%, 18.60% vs. 4.65%, 88.37% vs. 62.79%, 55.81% vs. 30.23%, 34.88% vs. 13.95%), had higher CTSI score (6.60±1.61 vs. 5.77±1.32), lower hemoglobin level at discharge (g/L: 102.30±18.78 vs. 110.72±16.68), and a lower proportion of etiological interventions after discharge (34.88% vs. 67.44%), the differences were statistically significant (all P < 0.05). Multivariate Logistic regression analysis showed that hypertension [odds ratio (OR) = 4.814, 95% confidence interval (95%CI) was 1.196-19.378], initial serum calcium < 2.0 mmol/L (OR = 6.688, 95%CI was 1.321-33.873) and initial blood glucose > 11.1 mmol/L (OR = 6.473, 95%CI was 1.399-29.950) were risk factors for long-term quality of life in cured SAP patients (all P < 0.05), while post discharge prophylactic intervention was a protective factor for long-term quality of life (OR = 0.092, 95%CI was 0.020-0.425, P < 0.01).
CONCLUSIONS
Cured SAP patients have varying degrees of impaired secretion function and the possibility of recurrence of acute pancreatitis. Hypertension, initial serum calcium < 2.0 mmol/L and blood glucose > 11.1 mmol/L are independent influencing factors for low long-term quality of life in cured SAP patients. Prevention and intervention targeting the etiology of pancreatitis after discharge can improve the long-term quality of life of cured SAP patients.
Humans
;
Quality of Life
;
Retrospective Studies
;
Pancreatitis/therapy*
;
Patient Discharge
;
Male
;
Female
;
Middle Aged
;
APACHE
;
Adult
;
Acute Disease
;
Aged
4.Effects of sauchinone regulating KLF5-EphA2 pathway on the growth and immune escape of hepatocellular carcinoma cells
Shan WANG ; Jinliang WAN ; Zhonghua ZHAO ; Jing YANG ; Jie ZHAO ; Yanzhang HAO
The Journal of Practical Medicine 2025;41(10):1480-1486
Objective To investigate the effects of sauchinone(Sch)on the growth and immune escape of liver cancer cells by regulating the Krüppel like factor 5(KLF5)-erythropoietin-producing hepatocellular receptor A2(EphA2)pathway.Methods Huh-7 cells were grouped into liver cancer group,Sch low,medium,and high-dose groups(Sch-L group,Sch-M group,Sch-H group),Sch-H+KLF5 activator negative control group(OE-NC),and Sch-H+KLF5 activator(OE-KLF5)group.5-bromo-2-deoxyuracil(EdU)staining,CCK-8,scratch assay,and Transwll were used to detect the proliferation,migration,and invasion of Huh-7 cells,respectively.Western blot was applied to detect proliferative cell nuclear antigen(PCNA),migration invasion enhancer(MIEN1),matrix metalloproteinase-2(MMP-2),programmed death receptor ligand 1(PD-L1),KLF5,and EphA2 proteins in Huh-7 cells.The Huh-7 cells in above 6 groups were co cultured with activated CD8+T cells in a 96 well plate and named as liver cancer co culture group,Sch-L co culture group,Sch-M co culture group,Sch-H co culture group,Sch-H+OE-NC co culture group,and Sch-H+OE-KLF5 co culture group.The killing rate of CD8+T cells in the co culture system and the levels of interferon-γ(IFN-γ),interleukin-4(IL-4),and tumor necrosis factor-α(TNF-α)in the supernatant were detected.Results Compared with the liver cancer group,the EdU positive rate,OD450 value,cell invasion number,and PCNA,MIEN1,MMP-2,PD-L1,KLF5,and EphA2 proteins in the Sch-L,Sch-M,and Sch-H groups reduced,the migration distance shorten(P<0.05).Compared with the Sch-H group and Sch-H+OE-NC group,the EdU positive rate,OD450 value,cell invasion number,and PCNA,MIEN1,MMP-2,PD-L1,KLF5,and EphA2 proteins in the Sch-H+OE-KLF5 group increased,the migration distance prolonged(P<0.05).Compared with the liver cancer co culture group,the killing rate of CD8+T cells on Huh-7 cells and the levels of IFN-γ,TNF-α,and IL-4 in the supernatant in the Sch-L co culture group,Sch-M co culture group,and Sch-H co culture group increased(P<0.05).Compared with the Sch-H co culture group and the Sch-H+OE-NC co culture group,the killing rate of CD8+T cells on Huh-7 cells and the levels of IFN-γ,TNF-α,and IL-4 in the supernatant in the Sch-H+OE-KLF5 co culture group decreased(P<0.05).Conclusion Sch may inhibit growth and immune escape of liver cancer cells by inhibiting the KLF5-EphA2 pathway.
5.Effects of sauchinone regulating KLF5-EphA2 pathway on the growth and immune escape of hepatocellular carcinoma cells
Shan WANG ; Jinliang WAN ; Zhonghua ZHAO ; Jing YANG ; Jie ZHAO ; Yanzhang HAO
The Journal of Practical Medicine 2025;41(10):1480-1486
Objective To investigate the effects of sauchinone(Sch)on the growth and immune escape of liver cancer cells by regulating the Krüppel like factor 5(KLF5)-erythropoietin-producing hepatocellular receptor A2(EphA2)pathway.Methods Huh-7 cells were grouped into liver cancer group,Sch low,medium,and high-dose groups(Sch-L group,Sch-M group,Sch-H group),Sch-H+KLF5 activator negative control group(OE-NC),and Sch-H+KLF5 activator(OE-KLF5)group.5-bromo-2-deoxyuracil(EdU)staining,CCK-8,scratch assay,and Transwll were used to detect the proliferation,migration,and invasion of Huh-7 cells,respectively.Western blot was applied to detect proliferative cell nuclear antigen(PCNA),migration invasion enhancer(MIEN1),matrix metalloproteinase-2(MMP-2),programmed death receptor ligand 1(PD-L1),KLF5,and EphA2 proteins in Huh-7 cells.The Huh-7 cells in above 6 groups were co cultured with activated CD8+T cells in a 96 well plate and named as liver cancer co culture group,Sch-L co culture group,Sch-M co culture group,Sch-H co culture group,Sch-H+OE-NC co culture group,and Sch-H+OE-KLF5 co culture group.The killing rate of CD8+T cells in the co culture system and the levels of interferon-γ(IFN-γ),interleukin-4(IL-4),and tumor necrosis factor-α(TNF-α)in the supernatant were detected.Results Compared with the liver cancer group,the EdU positive rate,OD450 value,cell invasion number,and PCNA,MIEN1,MMP-2,PD-L1,KLF5,and EphA2 proteins in the Sch-L,Sch-M,and Sch-H groups reduced,the migration distance shorten(P<0.05).Compared with the Sch-H group and Sch-H+OE-NC group,the EdU positive rate,OD450 value,cell invasion number,and PCNA,MIEN1,MMP-2,PD-L1,KLF5,and EphA2 proteins in the Sch-H+OE-KLF5 group increased,the migration distance prolonged(P<0.05).Compared with the liver cancer co culture group,the killing rate of CD8+T cells on Huh-7 cells and the levels of IFN-γ,TNF-α,and IL-4 in the supernatant in the Sch-L co culture group,Sch-M co culture group,and Sch-H co culture group increased(P<0.05).Compared with the Sch-H co culture group and the Sch-H+OE-NC co culture group,the killing rate of CD8+T cells on Huh-7 cells and the levels of IFN-γ,TNF-α,and IL-4 in the supernatant in the Sch-H+OE-KLF5 co culture group decreased(P<0.05).Conclusion Sch may inhibit growth and immune escape of liver cancer cells by inhibiting the KLF5-EphA2 pathway.
6.Role of lateral habenula and its associated neural circuitry projections in pain regulation
Yanjuan REN ; Dongxu WANG ; Ya CAO ; Yuxuan ZHANG ; Lu QIAN ; Danru WU ; Zhonghua LI ; Ling ZHANG ; Yu SHEN ; He LIU
Chinese Journal of Behavioral Medicine and Brain Science 2025;34(5):465-469
Pain modulation encompasses a complex neurobiological process, in which the lateral habenula (LHb) plays a crucial role in integrating, regulating and modulating pain signals. It is also involved in pain-related memory functions associated with perception, transmission and regulation of pain. Furthermore, the LHb collaborates with structures such as the spinal dorsal horn, forebrain, and amygdala to form an essential neural circuit that contributes to sensitization, development of tolerance, and adaptation processes related to pain. However, there remains limited understanding regarding the specific roles and interactions among different neuron subtypes within the LHb concerning pain regulation. Additionally, further investigation is warranted to explore functional changes and plasticity within both the LHb and its associated neural circuits in chronic pain models. Future research endeavors should utilize advanced neuroimaging techniques alongside optogenetics and gene editing technologies to elucidate intricate neural circuits, cellular architecture, and molecular mechanisms governing LHb function in pain regulation. In conclusion, this paper aims to comprehensively review existing literature on the involvement of the LHb and its neural circuits in modulating pain, thereby enhancing our understanding of their neurobiological mechanisms while providing novel targets for precise therapeutic strategies aimed at alleviating pain.
7.Construction of a risk prediction model for failure of proximal femoral nail antirotation fixation in intertrochanteric fractures
Zesong TU ; Daxing XU ; Hongbin LUO ; Yusheng WANG ; Xinglun FENG ; Zhonghua PENG ; Shaolong DU
Chinese Journal of Tissue Engineering Research 2025;29(27):5845-5853
BACKGROUND:Intertrochanteric femoral fractures are the main type of fragility fracture in the elderly,and proximal femoral nail antirotation is the preferred surgical option,but the factors associated with postoperative internal fixation failure are controversial.OBJECTIVE:A new"three-column"classification of intertrochanteric femoral fractures was proposed by evaluating patients'imaging data preoperatively and analyzing its interaction with postoperative internal fixation failure.A risk prediction model was developed and validated by using numerical algorithms,which facilitates clinicians to identify and intervene in high-risk patients preoperatively.METHODS:Patients with intertrochanteric femur fractures in Sanshui Branch of Foshan Hospital of Traditional Chinese Medicine between June 2012 and June 2022 were selected.The patients were divided into the internal fixation failure group and the internal fixation maintenance group according to whether they had internal fixation failure after surgery.According to the preoperative radiographs,the proximal femur was divided into three columns:the medial column,the lateral column,and the middle column.Each column had different subgroups.The relationship between the morphological characteristics of the"three columns"and the failure of proximal femoral nail antirotation internal fixation was analyzed,and the independent risk factors for internal fixation failure were screened out by single and then multifactorial logistic regression analyses.A risk prediction model was constructed according to the independent risk factors using R language software.The Bootstrap method was used to resample 1 000 times.The area under the curve,calibration curve,and clinical decision curve were used to evaluate the differentiation,calibration ability,and clinical application value of the model.The Youden index was used to determine the optimal risk cut-off value of the prediction model,according to which the patients were divided into high and low risk groups.The stability and extensibility of the model were evaluated according to the accuracy of its risk prediction ability.RESULTS AND CONCLUSION:(1)The four independent risk factors for postoperative internal fixation failure after surgery were predicted using the"three-column"typing system:medial column(comminuted fracture of the lesser trochanter and femoral talar)[odds ratio=5.385,95%CI(1.961,14.782),P=0.001],medial column(chimney type)[odds ratio=2.893,95%CI(1.167,7.173),P=0.022],lateral column(lateral wall thickness<20.5 mm)[odds ratio=2.804,95%CI(1.078,7.297),P=0.035]and lateral column(lateral wall fracture)[odds ratio=4.278,95%CI(1.670,10.959),P=0.012].(2)The constructed risk prediction model showed good discrimination and accuracy[area under the receiver operating characteristic curve=0.852,95%CI(0.837,0.922)].The calibration curve showed good agreement between the model-predicted risk and the actual risk of occurrence.(3)The clinical decision curve suggested that the model had good clinical applicability when the risk threshold probability was in the range of 0.2-0.82.The risk probability of 28%was the optimal threshold for risk stratification of the model,and the predictive performance of the model was better in patients with different risk groups.(4)The"three-column"typing system constructs a predictive model to calculate the risk probability of postoperative internal fixation failure in patients with intertrochanteric femoral fractures.This method is accurate,simple,and easy to apply clinically,and can be used as a digital tool to guide personalized clinical treatment.
8.Impacts of paeoniflorin on inflammation and NF-κB/NLRP3 signal pathway in knee osteoarthritis model rats
Zhongbo ZHANG ; Fuzeng ZHENG ; Shangzeng WANG ; Dongliang SHI ; Zhonghua GUO ; Yunfei WANG ; Zhentao WANG
Chinese Journal of Immunology 2025;41(8):1826-1832
Objective:To investigate impacts of paeoniflorin on inflammation and nuclear transcription factor-κB(NF-κB)/nucleotide-binding oligomeric domain-like receptor protein 3(NLRP3)signal pathway in knee osteoarthritis(KOA)model rats.Meth-ods:SD rats were randomly grouped into model group,glucosamine group,paeoniflorin low-dose group,paeoniflorin high-dose group,paeoniflorin high-dose+phorbol ester(PMA)(NF-κB activator)group,with 10 rats in each group,another 10 rats were regarded as control group,and only knee joint capsule was cut,after treatment with glucosamine,paeoniflorin and PMA,joint pain symptoms of rats were detected by mechanical stimulation method and thermal radiation method;knee joint width,joint swelling and synovial thick-ness were measured;HE staining was applied to detect joint tissue structure and morphology of rats in each group;levels of inflamma-tory factors IL-1β,monocyte chemoattractant protein 1(MCP-1)and IL-9 in joint fluid and serum of rats were detected by ELISA;and Western blot was applied to detect expressions of NF-κB/NLRP3 signal pathway related proteins in rat joint tissue.Results:Com-pared with control group,joint tissue structure of model group was significantly damaged,mechanical pain threshold and thermal sen-sitivity pain threshold were significantly lower(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expression of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were higher(P<0.05).Compared with model group,joint tissue damage of rats in glucosamine group,paeoniflorin low-dose group and paeoniflorin high-dose group was reduced,mechanical pain threshold and thermal sensitivity pain threshold were higher(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expressions of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were lower(P<0.05);joint tissue injury of rats in paeoniflorin high-dose group was further reduced compared with paeoniflorin low-dose group,mechanical pain threshold and thermal sensitivity pain threshold were further higher(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expression of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were further lower(P<0.05).Compared with paeoniflorin high-dose group,joint tissue damage of rats in paeoniflorin high-dose+PMA group was increased,mechanical pain threshold and thermal sensitivity pain threshold were lower(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expression of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were higher(P<0.05);there was no significant change in all indexes of rats in glucosamine group(P>0.05).Conclusion:Paeoniflorin can play an anti-inflammatory role by down-regulating the NF-κB/NLRP3 signal pathway,thus alleviating joint injury and joint pain symptoms of KOA rats.
9.Role of lateral habenula and its associated neural circuitry projections in pain regulation
Yanjuan REN ; Dongxu WANG ; Ya CAO ; Yuxuan ZHANG ; Lu QIAN ; Danru WU ; Zhonghua LI ; Ling ZHANG ; Yu SHEN ; He LIU
Chinese Journal of Behavioral Medicine and Brain Science 2025;34(5):465-469
Pain modulation encompasses a complex neurobiological process, in which the lateral habenula (LHb) plays a crucial role in integrating, regulating and modulating pain signals. It is also involved in pain-related memory functions associated with perception, transmission and regulation of pain. Furthermore, the LHb collaborates with structures such as the spinal dorsal horn, forebrain, and amygdala to form an essential neural circuit that contributes to sensitization, development of tolerance, and adaptation processes related to pain. However, there remains limited understanding regarding the specific roles and interactions among different neuron subtypes within the LHb concerning pain regulation. Additionally, further investigation is warranted to explore functional changes and plasticity within both the LHb and its associated neural circuits in chronic pain models. Future research endeavors should utilize advanced neuroimaging techniques alongside optogenetics and gene editing technologies to elucidate intricate neural circuits, cellular architecture, and molecular mechanisms governing LHb function in pain regulation. In conclusion, this paper aims to comprehensively review existing literature on the involvement of the LHb and its neural circuits in modulating pain, thereby enhancing our understanding of their neurobiological mechanisms while providing novel targets for precise therapeutic strategies aimed at alleviating pain.
10.Forensic application evaluation of ssDNA library construction-hybridization capture DNA methylation sequencing in height inference
Zhonghua WANG ; Qian WANG ; Guangping FU ; Junyan WANG ; Lihong FU ; Bin CONG ; Shujin LI
Chinese Journal of Forensic Medicine 2025;40(1):33-39,55
Objective To evaluate the sensitivity,repeatability,anti-degradation ability,in vivo temporal stability,and tissue specificity of the DNA methylation sequencing panel constructed in our previous study for height inference,so as to provide a reference for forensic application.Methods Sensitivity:different initial template quantities(50 ng,40 ng,30 ng,20 ng)were set for sequencing.Repeatability:DNA from the same sample was sequenced three times.Anti-degradation ability:whole blood was used to make blood stains,and DNA was extracted and sequenced at 0,3,6 and 9 months,respectively.In vivo temporal stability:the blood was collected at 0,3,6,and 9 months for sequencing.Tissue specificities:published data and findings were used to analyze the tissue specificities of CpGs in the panel.Results The sensitivity test showed that the initial template quantities of 20 ng detected all the CpG sites and still obtained accurate prediction results.The results of the three repeated predictions of the same sample are stable,and the differences are mainly due to the randomness of the DNN model,indicating good detection repeatability.A complete methylation profile was obtained for the blood stains left at room temperature for 9 months,and the predicted results showed a small range of fluctuations.The three samples were predicted to fluctuate within a range of 1.5 cm or less over nine months.Tissue-specific analyses showed a high correlation between blood and saliva,but can not apply to other tissues.Conclusion The DNA methylation detection system we developed in our previous study has good sensitivity,repeatability,anti degradation ability,in vivo time stability,as well as strong tissue specificity,making it suitable for height inference of blood samples.This supports the feasibility of using targeted DNA methylation analysis on whole blood samples to infer height in the field of forensic science.

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