BACKGROUND:Bone marrow mesenchymal stem cells play a crucial role in treatment of diseases,such as femoral head necrosis,and the therapeutic efficacy is closely related to the quality of the cells.The empowerment of cells has become a research focus.OBJECTIVE:To investigate the effects of hypoxia mimetic dimethylglyoxal glycine pretreatment on mitophagy and differentiation capacity of human bone marrow mesenchymal stem cells.METHODS:Bone marrow mesenchymal stem cells were extracted from the bone marrow of patients' iliac crest and cultured in vitro to the third passage.The cells were treated with dimethylglyoxal glycine at 0,10,50,and 100 μmol/Lfor 24 hours,followed by the replacement with an osteogenic induction differentiation medium,which constituted the pretreatment group.The continuous treatment group was cultured directly in osteogenic induction medium containing 0,10,50,and 100 μmol/L dimethylglyoxal glycine after cell adhesion.After 7 days of induction,alkaline phosphatase staining was performed to select the most favorable conditions for osteogenic differentiation for subsequent experiments,with normally cultured bone marrow mesenchymal stem cells serving as the control group.Alkaline phosphatase staining,alkaline phosphatase activity,oil red O staining,and related RT-qPCR were used to evaluate the osteogenic and adipogenic differentiation differences of bone marrow mesenchymal stem cells between the two groups.MitoSox staining was used to detect mitochondrial reactive oxygen species levels.Mito-tracker and Lyso-tracker staining were used to detect the co-localization of mitochondria and lysosomes.The fluorescent probe JC-1 was used to measure mitochondrial membrane potential.RESULTS AND CONCLUSION:Alkaline phosphatase staining indicated that the most beneficial treatment for bone marrow mesenchymal stem cell osteogenesis was pretreatment with 10 μmol/L dimethylglyoxal glycine for 24 hours.Compared with the control group,the experimental group showed enhanced alkaline phosphatase staining expression,increased alkaline phosphatase activity and osteogenic gene expression,reduced lipid droplet formation and adipogenic gene expression as indicated by oil red O staining,decreased mitochondrial reactive oxygen species production,increased co-localization of mitochondria and lysosomes,and elevated mitochondrial membrane potential.The results suggest that 10 μmol/L dimethylglyoxal glycine pretreatment can promote osteogenic differentiation of bone marrow mesenchymal stem cells,inhibit adipogenic differentiation,and enhance mitophagy.

BACKGROUND:Bone marrow mesenchymal stem cells play a crucial role in treatment of diseases,such as femoral head necrosis,and the therapeutic efficacy is closely related to the quality of the cells.The empowerment of cells has become a research focus.OBJECTIVE:To investigate the effects of hypoxia mimetic dimethylglyoxal glycine pretreatment on mitophagy and differentiation capacity of human bone marrow mesenchymal stem cells.METHODS:Bone marrow mesenchymal stem cells were extracted from the bone marrow of patients' iliac crest and cultured in vitro to the third passage.The cells were treated with dimethylglyoxal glycine at 0,10,50,and 100 μmol/Lfor 24 hours,followed by the replacement with an osteogenic induction differentiation medium,which constituted the pretreatment group.The continuous treatment group was cultured directly in osteogenic induction medium containing 0,10,50,and 100 μmol/L dimethylglyoxal glycine after cell adhesion.After 7 days of induction,alkaline phosphatase staining was performed to select the most favorable conditions for osteogenic differentiation for subsequent experiments,with normally cultured bone marrow mesenchymal stem cells serving as the control group.Alkaline phosphatase staining,alkaline phosphatase activity,oil red O staining,and related RT-qPCR were used to evaluate the osteogenic and adipogenic differentiation differences of bone marrow mesenchymal stem cells between the two groups.MitoSox staining was used to detect mitochondrial reactive oxygen species levels.Mito-tracker and Lyso-tracker staining were used to detect the co-localization of mitochondria and lysosomes.The fluorescent probe JC-1 was used to measure mitochondrial membrane potential.RESULTS AND CONCLUSION:Alkaline phosphatase staining indicated that the most beneficial treatment for bone marrow mesenchymal stem cell osteogenesis was pretreatment with 10 μmol/L dimethylglyoxal glycine for 24 hours.Compared with the control group,the experimental group showed enhanced alkaline phosphatase staining expression,increased alkaline phosphatase activity and osteogenic gene expression,reduced lipid droplet formation and adipogenic gene expression as indicated by oil red O staining,decreased mitochondrial reactive oxygen species production,increased co-localization of mitochondria and lysosomes,and elevated mitochondrial membrane potential.The results suggest that 10 μmol/L dimethylglyoxal glycine pretreatment can promote osteogenic differentiation of bone marrow mesenchymal stem cells,inhibit adipogenic differentiation,and enhance mitophagy.

Dendritic cells (DCs) serve as the primary antigen-presenting cells in autoimmune diseases, like rheumatoid arthritis (RA), and exhibit distinct signaling profiles due to antigenic diversity. Type II collagen (CII) has been recognized as an RA-specific antigen; however, little is known about CII-stimulated DCs, limiting the development of RA-specific therapeutic interventions. In this study, we show that CII-stimulated DCs display a preferential gene expression profile associated with migration, offering a new perspective for targeting DC migration in RA treatment. Then, saikosaponin D (SSD) was identified as a compound capable of blocking CII-induced DC migration and effectively ameliorating arthritis. Optineurin (OPTN) is further revealed as a potential SSD target, with Optn deletion impairing CII-pulsed DC migration without affecting maturation. Function analyses uncover that OPTN prevents the proteasomal transport and ubiquitin-dependent degradation of C-C chemokine receptor 7 (CCR7), a pivotal chemokine receptor in DC migration. Optn-deficient DCs exhibit reduced CCR7 expression, leading to slower migration in CII-surrounded environment, thus alleviating arthritis progression. Our findings underscore the significance of antigen-specific DC activation in RA and suggest OPTN is a crucial regulator of CII-specific DC migration. OPTN emerges as a promising drug target for RA, potentially offering significant value for the therapeutic management of RA.

OBJECTIVE: To explore the integrated traditional Chinese and Western medicine treatment plan for ankylosing spondylitis complicated with lower cervical spine fracture and dislocation, adopt the treatment plan of preoperative continuous traction, intraoperative prizing reduction combined with anterior long-segment plate-screw and posterior short-segment pedicle screw-rod system internal fixation, and evaluate its surgical efficacy and clinical application value. METHODS: From June 2018 to September 2022, 7 male patients with ankylosing spondylitis complicated with lower cervical spine fractures were admitted, aged 43 to 65 years old. Among them, there was 1 case of C_3,4 fracture, 1 case of C_4,5 fracture, 1 case of C_6,7 fracture, and 4 cases of C_5,6 fracture, all of which were fracture and dislocation. All patients received preoperative continuous skull traction, and intraoperative prizing reduction combined with anterior long-segment plate-screw and posterior short-segment pedicle screw-rod system internal fixation. The Neck Disability Index (NDI), Japanese Orthopedic Association (JOA) score, and Frankel scale were used to evaluate the neurological function and quality of life before and after surgery. The visual analogue scale (VAS) was used to evaluate neck and limb pain. The operation time, blood loss, hospital stay, and surgery-related complications were recorded. RESULTS: All 7 patients were followed up for 6 to 24 months after surgery. The operation time of the 7 patients ranged from 300 to 480 minutes, the blood loss ranged from 300 to 1000 ml, and the hospital stay ranged from 8 to 25 days. The preoperative NDI of the 7 patients ranged from 25% to 42%, which decreased to 12% to 30% at 1 week after surgery and 5% to 25% at the last follow-up. The preoperative JOA score ranged from 8 to 13 points, which increased to 12 to 15 points at 1 week after surgery and 13 to 16 points at the last follow-up. The preoperative VAS ranged from 6 to 8 points, which decreased to 2 to 4 points at 1 week after surgery and 0 to 3 points at the last follow-up. Regarding the Frankel grade of neurological function, 2 patients were grade C before surgery and recovered to grade D at the last follow-up after surgery, and the remaining patients recovered to grade E at the last follow-up after surgery. There were 3 cases of pressure ulcers, including 1 case of intraoperative pressure ulcer, 1 case of cervical cerebrospinal fluid leakage, 1 case of screw loosening, and 1 case of aggravated fracture dislocation due to preoperative traction. CONCLUSION Preoperative cervical traction combined with intraoperative prizing reduction and anterior long-segment plate combined with posterior short-segment pedicle screw internal fixation provides a safe and effective surgical option for ankylosing spondylitis complicated with lower cervical spine fracture and dislocation, which can minimize surgical trauma and improve clinical efficacy. However, this study has a small sample size and a short follow-up time for some patients, so further verification with large-sample and long-term follow-up data is still needed.
Humans ; Male ; Adult ; Middle Aged ; Fracture Fixation, Internal/methods* ; Spondylitis, Ankylosing/surgery* ; Cervical Vertebrae/surgery* ; Spinal Fractures/surgery* ; Traction ; Aged ; Joint Dislocations/surgery*

OBJECTIVE: To investigate the effects of histone deacetylase (HDAC) levels on the proliferation and apoptosis of Burkitt lymphoma cells, and the changes in related signaling molecules in the PI3K/AKT/mTOR signaling pathway, so as to explore the pathogenesis of Burkitt lymphoma. METHODS: HDAC levels in Burkitt lymphoma were detected by RT-PCR and Western blot. CA46 and RAJI cells were treated with the HDAC selective inhibitor VPA. CCK8 assay was used to detect the proliferation ability of cells. Western Blot was used to measure the expression of apoptosis-related proteins, PI3K/AKT/mTOR signaling pathway proteins and their phosphorylation levels. RESULTS: The expression levels of classⅠ HDAC in Burkitt lymphoma were higher than those in normal cells, and the HDAC1 inhibitor VPA could inhibit the proliferation of CA46 and RAJI cells. VPA decreased HDAC expression in CA46 and RAJI cells, inhibited the phosphorylation of PI3K/AKT/mTOR pathway molecules AKT and p70S6K, increased the expression of apoptotic proteins Cleaved Caspase-3, Cleaved Caspase-8, Cleaved Caspase-9 and Bax, and decreased the expression of anti-apoptotic proteins Bcl-2 and PARP. CONCLUSION Inhibition of HDAC activity can Attenuate the proliferation of Burkitt lymphoma cells and induce apoptosis by inhibiting the PI3K/AKT/mTOR signaling pathway activity.
Humans ; Burkitt Lymphoma/pathology* ; Apoptosis ; Cell Proliferation ; Signal Transduction ; Proto-Oncogene Proteins c-akt/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Cell Line, Tumor ; Histone Deacetylases/metabolism* ; TOR Serine-Threonine Kinases/metabolism* ; Histone Deacetylase Inhibitors/pharmacology* ; Phosphorylation

Objective:To investigate the dynamic characteristics of human papillomavirus (HPV) genomic integration during cervical lesion progression and the clinical value of HPV integration detection in stratify HPV-positive women, and to explore its molecular mechanisms in cervical carcinogenesis.Methods:A prospective cohort study was designed to enroll high-risk HPV (HR-HPV) positive women who underwent cervical cancer screening in Drum Tower Hospital Affiliated to Nanjing University Medical School and Nanjing Maternity and Child Health Care Hospital from July 2022 to July 2024. Cervical exfoliated cells samples were collected, and HPV whole genome targeted capture and high-throughput sequencing technology were used. The HPV integration patterns, host gene functional region distribution and pathway enrichment characteristics of 157 samples with different cervical lesions grades were analyzed, including 31 cases of normal cervix, 40 cases of cervical intraepithelial neoplasia (CIN) Ⅰ, 32 cases of CIN Ⅱ, 42 cases of CIN Ⅲ, and 12 cases of cervical cancer.Results:HR-HPV integration was detected in 80.2% (126/157) of the 157 HR-HPV positive samples. The incidence of HR-HPV integration in cervical cancer patients was 12/12, which was higher than that in normal women (77%, 24/31). The incidence of HPV16 integration was significantly higher in high-grade lesions, and the incidence of HPV16 integration was 43% (18/42) in CIN Ⅲ patients and 8/12 in cervical cancer patients ( P<0.001). A total of 14 438 integration events were detected in 126 samples with HPV integration. The integration sites were mainly distributed in the host intergenic region (51.0%, 7 359/14 438) and intronic region (38.1%, 5 494/14 438), and the integration frequency of viral L1 gene was the highest (28.4%, 4 498/16 781). Functional enrichment analysis showed that HPV integration-related host genes were significantly enriched in transport of small molecules,cyclic guanosine monophosphate (cGMP)-protein kinase G (PKG) signaling pathway, and purine ribonucleotide biosynthetic process, which synergistically drove carcinogenesis through multiple mechanisms. Conclusions:HPV integration events are significantly associated with the progression of cervical lesions. HPV integrated detection based on cervical exfoliated cells is expected to optimize the current screening strategy, reduce excessive intervention of HPV positive women and facilitate their accurate triage management.

INTRODUCTION: Obstructive sleep apnoea (OSA) is common in Singapore, with moderate to severe OSA affecting around 30% of residents. These consensus statements aim to provide scientifically grounded recommendations for the management of OSA, standar-dise the management of OSA in Singapore and promote multidisciplinary collaboration. METHOD: An expert panel, which was convened in 2024, identified several areas of OSA management that require guidance. The expert panel reviewed the current literature and developed consensus statements, which were later independently voted on using a 3-point Likert scale (agree, neutral or disagree). Consensus (total ratings of agree and neutral) was set a priori at ≥80% agreement. Any statement not reaching consensus was excluded. RESULTS: The final consensus included 49 statements that provide guidance on the screening, diagnosis and management of adults with OSA. Additionally, 23 statements on the screening, diagnosis and management of paediatric OSA achieved consensus. These 72 consensus statements considered not only the latest clinical evidence but also the benefits and harms, resource implications, feasibility, acceptability and equity impact of the recommendations. CONCLUSION The statements presented in this paper aim to guide clinicians based on the most updated evidence and collective expert opinion from sleep specialists in Singapore. These recommendations should augment clinical judgement rather than replace it. Management decisions should be individualised, taking into account the patient's clinical characteristics, as well as patient and caregiver concerns and preferences.
Humans ; Sleep Apnea, Obstructive/diagnosis* ; Singapore ; Consensus ; Adult

AIM:To study the protective effect of transient receptor potential vanilic acid subtype 1(TRPV1)agonist capsaicin(CAP)on traumatic blood loss shock rats,and to further explore its possible mechanism by network pharmacology.METHODS:Forty-five SD rats were divided into 5 groups by random number table method:normal group,shock group,lactated Ringer's solution(LR)group,CAP pretreatment(single administration before shock)group,CAP pre-final administration(twice administration before and after shock)group,with 9 rats in each group for survival observation.Then 32 SD rats were divided into 4 groups according to the results of survival experiment:normal group,shock group,LR group,CAP pre-final administration group,with 8 rats in each group for blood pressure,hemodynamics,arterial blood gas,vascular reactivi-ty and hepaticand renal blood flow.At the same time,the potential mechanism of CAP in the treat-ment of traumatic hemorrhagic shock was investi-gated by network pharmacology.Furthermore,ap-ply the dataset to validate and analyse the diagnos-tic value of the hub genes.RESULTS:Rats in shock group died within hours of the completion of the shock model,and the mean survival time was 1.25(0.42,6.21)h.LR resuscitation could improve the survival of rats to some extent.The survival rate and survival time of rats in the CAP pretreatment group were slightly increased as compared with the LR group,while twice administration of CAP be-fore and after shock(CAP pre-final administration)resulted in better outcomes than LR resuscitation alone.Further results indicated that CAP pre-final administration significantly reduced the blood lac-tic acid level,improved the vasoconstrictive and di-astolic reactivity,and increased the liver and kidney blood flow of shock rats as compared with LR group.The improvement of hemodynamics and blood gas indexes in CAP group was slightly higher than LR group,but there was no statistical signifi-cance.A total of 37 genes related to CAP anti-trau-matic hemorrhage shock were obtained by net-work pharmacology.KEGG enrichment analysis showed that the Ca ion signaling pathway and Ras signaling pathway were significantly enriched.Vali-dation of the dataset showed that the expression levels of CXCR4,NF-kB1,GFPA and NTF3 hub gene were significantly different in the normal and shock groups,and that CXCR4 has a high diagnostic value for traumatic haemorrhagic shock.CONCLUSIONS:CAP,the TRPV1 agonist,significantly improved vas-cular function,increased organ blood flow,and cor-rected the lactic acidosis in rats with traumatic hemorrhagic shock,thus markedly improved the survival outcomes.The mechanism may be related to Ca ion signal pathway and Ras signal pathway.CXCR4,NF-kB1,GFPA and NTF3 may be having an important role in it.

Aiming at the problems of tongue diagnosis instruments relying on manual operation,uneven illumination,color distortion and lack of dynamic information,an automated operation black box tongue diagnosis instrument based on machine vision is proposed.This instrument utilizes machine vision technology and image processing algorithms to achieve automatic positioning and automatic acquisition of tongue manifestations.High-resolution camera technology enables the acquisition of 3-second dynamic videos and high-quality static images.The performance test results show that,based on the dual-camera geometric projection calibration algorithm,the average time for tongue recognition is less than 0.15 seconds.The diffuse reflection light source improves the gray uniformity to 76.74%,and the maximum color difference in color reproduction degree is 19,which is better than the industry standard value of 25.The developed tongue diagnosis instrument will promote the objective,efficient and precise acquisition of tongue diagnosis in traditional Chinese medicine.

BACKGROUND:The development of calcium phosphate bone cement is limited due to its poor mechanical properties and weak osteogenic ability.Silicate bioactive glass is highly favored due to its excellent biological activity and osteogenic ability.Simultaneously,fiber structures can enhance the mechanical strength of materials.OBJECTIVE:To investigate the mechanical properties,biocompatibility,and osteogenic effect of silicate bioactive glass fiber composite calcium phosphate bone cement.METHODS:Different mass percentages(0％,10％,and 20％)of silicate bioactive glass fiber were added to the solid phase of calcium phosphate bone cement,mixed with the liquid phase and cured for 48 hours to obtain silicate bioactive glass fiber composite calcium phosphate bone cement.The mechanical properties,setting time,and ion precipitation of the cement were characterized.The three groups of bone cement extracts were co-cultured with MC3T3-E1 cells.The cell compatibility of the materials was evaluated by CCK-8 assay,live/dead staining,and phalloidin staining.After osteogenic induction,the osteogenic induction ability of the materials was evaluated by alkaline phosphatase staining,alizarin red staining,RUNX2 immunofluorescence staining,and RT-PCR.RESULTS AND CONCLUSION:(1)With the increase of silicate bioactive glass fiber content,the compressive strength and flexural strength of bone cement increased,and the setting time was prolonged.When bone cement was immersed in simulated body fluid,the precipitation of silicon ions,calcium ions,and phosphorus ions could be detected.Moreover,with the increase of silicate bioactive glass fiber content,the mass concentration of silicon ions and phosphorus ions released by bone cement increased,and the mass concentration of calcium ions decreased.(2)Live/dead staining and phalloidin staining results exhibited that silicate bioactive glass fiber composite calcium phosphate bone cement had no toxic effect on MC3T3-E1 cells.CCK-8 assay results showed that silicate bioactive glass fiber composite calcium phosphate bone cement could promote the proliferation of MC3T3-E1 cells.(3)With the increase of silicate bioactive glass fiber content in bone cement,the alkaline phosphatase activity and extracellular calcium deposition of MC3T3-E1 cells increased,the expression of RUNX2 protein increased,and the expression of alkaline phosphatase,osteocalcin,osteopontin,and RUNX2 mRNA expression increased.(4)The results indicate that silicate bioactive glass fibers can enhance the mechanical properties and osteogenic induction ability of calcium phosphate bone cement,among which 20％silicate bioactive glass fibers have a more obvious effect.