BACKGROUND:Intervertebral disc degeneration is clinically considered to be the main cause of low back pain,but due to the unclear pathogenesis of intervertebral disc degeneration,there is still a lack of effective means to delay the progression of the disease.Single-cell RNA sequencing technology can amplify and sequence mRNA at the single-cell level,reveal the gene expression intensity of a single cell,discover different cell subsets in tissues according to the heterogeneity of cells,study the pathogenesis of intervertebral disc degeneration at the molecular level,and provide a new theoretical basis for its early diagnosis and treatment. OBJECTIVE:To introduce the basic principles of single-cell RNA sequencing technology and review the research progress of single-cell RNA sequencing technology in intervertebral disc degeneration in recent years. METHODS:A computer was used to search PubMed,Web of Science,CNKI and WanFang databases for the literature published from 2012 to 2022.Key words were"single-cell RNA sequencing,intervertebral disc degeneration,sequencing Technology"in Chinese and English.Duplicate,poor-quality and irrelevant articles were excluded;a total of 70 articles were eventually included. RESULTS AND CONCLUSION:(1)We identified new cell subsets such as homeostatic chondrocytes,hypertrophy chondrocyte-like nucleus pulposus cells and fibrous nucleus pulposus cells,identified the marker genes and transcription factors of these cell subsets,and described the functions,differentiation paths and cell fate of these cell subsets during the development and progression of intervertebral disc degeneration,and proposed the concept of progenitor nucleus pulposus cells.A cell subpopulation with progenitor nucleus pulposus cells properties was identified and its effectiveness in treating intervertebral disc degeneration was verified in mice.(2)Fibro chondrocyte-like annulus fibrosus cells and annulus fibrosus stem cells with both cartilage and fiber properties were identified,and a new type of composite hydrogel was prepared by combining fibrous cartilage inducers silk fibroin and hyaluronic acid in vitro.Experiments in mice demonstrated that this hydrogel could repair both annulus fibrosus tissue and cartilage matrix,and was remarkably effective in the treatment of intervertebral disc degeneration.(3)Regulatory chondrocytes were found in endplate cartilage.Two distinct fates in the progression of intervertebral disc degeneration were analyzed and the differential genes in the two fates were identified.Intercellular communication analysis indicated that regulatory chondrocytes interact with endothelial cells to promote angiogenesis.(4)Immune cells such as macrophages,T cells,myeloid progenitor cells and neutrophils were identified in the degenerated intervertebral disc tissues,demonstrating the existence of immune response during intervertebral disc degeneration.It was found that apolipoprotein induced the polarization of macrophages M1 and M2 subtypes,and this polarization process affected the activity of progenitor nucleus pulposus cells by amplifying the inflammatory response through the MIF signaling pathway.

Objective:To evaluate the short-term efficacy and safety of 3D printing patient-matched artificial vertebral body in clinical research and application.Methods:A total of 12 patients with spinal tumors were enrolled 7 males (58.33%) and 5 females (41.67%), aged from 18 to 65 years old in The First Affiliated Hospital of Air Force military Medical University (hereinafter referred to as Xijing Hospital) and Peking University people's Hospital from September 2021 to July 2022. The spinal vertebra defect were restored by using 3D printing patient-matched artificial vertebral body after tumor resection. All patients who accepted TES and 3D printing patient-matched artificial vertebral body implantation were included according to the inclusion and exclusion criteria. The bone interface fusion was evaluated by the imaging fusion criteria of Brantigan and Steffee at 3 and 6 months after operation, the curative effect was evaluated by comparing Japanese Orthopaedic Association (JOA) score at 3 and 6 months after operation, visual analogue scale (VAS) 3 months after operation and intervertebral height at 3 and 6 months after operation with those before operation, and the safety was evaluated by adverse event recording.Results:All 12 patients completed the operation successfully, and the operation sites were thoracic vertebrae in 6 cases (50%), thoracolumbar in 3 cases (25%) and lumbar vertebrae in 3 cases (25%). All patients were followed up. The mean follow-up time was 23.92±3.23 months (range, 19-29 months). No tumor recurrence or metastasis was observed during this period. All patients were followed up at 15 days, 3 months and 6 months after operation. During the 6-month follow-up, X ray results showed that interface of bone and the vertebral body were fused in all of the 12 patients, and the effective rate of fusion was 100%. The 95% confidence interval is calculated to be (75.6%-100%). Six months after operation, the improvement rate of JOA score was excellent in 10 cases, good in 1 case, poor in 1 case, and the excellent and good rate was 91.66%. The preoperative VAS score was 4.08 ±2.47, and during the 3-month follow-up, the VAS score was improved to 1.83 ±1.59. Compared with the preoperative VAS score, the difference was statistically significant ( t=2.635, P=0.023). The intervertebral height before operation, 15 days after operation, 3 months after operation and 6 months after operation were 32.75 (25.94, 68.20), 41.09 (30.55, 70.20), 40.70 (30.23, 67.83) and 40.74 (30.23, 67.08), respectively, and there was no statistically significant difference (χ 2=0.768, P=0.857). No implant-related adverse events occurred after operation. Conclusion:The 3D printing patient-matched artificial vertebral body used in this study has satisfactory short-term efficacy and safety in the reconstruction of spinal stability after spinal tumor resection.

AIM: To analyze the modeling characteristics and validation indexes of diabetic retinopathy model, analyze the shortcomings of the present animal experimental modeling, and provide a reference basis for the establishment of the standardization of the diabetic retinopathy model.METHODS: Literatures related to animal experiments on diabetic retinopathy were searched through the databases of CNKI, Wanfang, VIP and PubMed, and the experimental animal species, grade, gender, age, modeling method, modeling period, validation indexes, and other indexes were summarized and analyzed.RESULTS: The 275 papers that met the criteria were included. The animal models of diabetic retinopathy were mainly SD rats and Wistar rats, the sex of the experimental animals was mainly male, and the animal breeds were mostly of the SPF class. The age of most of the animals used was in the range of 6-8 weeks old; the modeling was based on those who established the type 1 diabetes model, mainly using STZ as the induction model. While the type 2 diabetes model was based on the high-fat, high-sugar diet combined with STZ. The modeling criteria were verified by detecting retinal morphology and structure, retinal vascularization, retinal function, and retinal cell apoptosis. In addition, the model was evaluated by detecting abnormal vascular proliferation, oxidative stress indicators, and inflammatory factor levels in retinal tissues, as well as abnormal vascular proliferation and inflammatory factor levels in aqueous humour, vitreous humor, and blood serum.CONCLUSION: Although the animal model of diabetic retinopathy has become a hotspot, the existing review is not comprehensive. Therefore, we summarized and analyzed the elements of the animal model through literature collation, including its characteristics and limitations, and providing methodological references for the establishment of the model, with a view to laying a solid foundation for the subsequent clinical and basic research of traditional Chinese medicine.

ObjectiveThis study aimed to evaluate the clinical efficacy of Ruyi Zhenbaowan（RYZBW）in the treatment of initial and early knee osteoarthritis （KOA） through a prospective multicenter，randomized，double-blind，and placebo-parallel controlled trial. MethodFrom October 13^th， 2021 to December 25^th， 2021， 240 KOA subjects meeting the acceptance criteria were enrolled in 15 sub-centers including Wangjing Hospital， Chinese Academy of Chinese Medical Sciences， and they were randomly divided into observation group and control group， with 120 cases in each group. The intervention measures for the observation group were RYZBW + health education， and the intervention measures for the control group were RYZBW placebo + health education. The intervention period in both groups was four weeks， and they were followed up for four weeks after the intervention. The primary outcome measure was the total score of Western Ontario and McMaster University Osteoarthritis Index score （WOMAC score）， and the secondary outcome measures were the response rate of visual scale （VAS） pain score， WOMAC sub item scores （joint pain， joint stiffness， and joint function）， quality of life （SF-12） score， and traditional Chinese medicine （TCM） syndrome score. Result（1） Efficacy evaluation. The marginal model results showed that the observation group was better than the control group in improving the WOMAC total score and WOMAC pain score in the treatment of KOA with RYZBW， and the difference was statistically significant （P<0.05）. There was no significant difference between the two groups in improving VAS score response rate， WOMAC function score， WOMAC stiffness score， SF12-PCS （quality of life-physical health） score， SF12-MCS （quality of life-mental health） score， and TCM syndrome score. （2） Subgroup analysis. ① In terms of VAS score response rate， the response rate of the observation group was higher than that of the control group for subjects with baseline VAS score of （4， 5］， and the difference was statistically significant （P<0.05）. ② In terms of TCM syndrome score， for subjects aged ［56， 60］ and ［61， 65］， the decrease in total TCM syndrome score in the observation group was better than that in the control group， and the difference was statistically significant （P<0.05）. ConclusionTibetan medicine RYZBW has good clinical efficacy in improving WOMAC total score， VAS score response rate， WOMAC pain score， WOMAC function score， and TCM syndrome score for patients with initial and early KOA， which can fill the lack of Tibetan medicine RYZBW in the treatment of KOA and make a demonstration study for the inheritance and development of ethnic medicine.

Type 2 diabetes mellitus （T2DM） is based on insulin resistance （IR） and insulin secretion deficiency， with the specific mechanisms still unclear. Current research involves mechanisms such as glycolipid toxicity， inflammatory response， oxidative stress damage， and mitochondrial dysfunction. Modern traditional Chinese medicine （TCM） scholars have named it "blood glucose collateral disease" based on the clinical characteristics and natural progression of T2DM. This condition is primarily manifested as abnormal blood sugar levels in the early stages， and as the disease progresses， it gradually causes widespread damage to the body's veins and collaterals， ultimately leading to lesions in vessels and collaterals. Among these， "spleen heat" （obesity type） is the most common clinical type of T2DM. The concept of "internal heat-induced elimination" runs through both the onset and complications of T2DM， with internal heat being a key factor in its pathogenesis. The clinical application of Dahuang Huanglian Xiexintang and its modifications has achieved significant therapeutic effects. This paper reviews the origins and treatment characteristics of Dahuang Huanglian Xiexintang， along with clinical application research and experimental studies related to T2DM treatment， involving mechanisms for regulating glucose and lipid metabolism disorders， improving IR， modulating inflammatory responses， combating oxidative stress damage， regulating autophagy-related signaling pathways， modulating intestinal flora， inhibiting pyroptosis， and alleviating endoplasmic reticulum stress， with the purpose to provide direction for further research on the prevention and treatment of T2DM and its related complications， to offer reference for developing Dahuang Huanglian Xiexintang as a rapid hypoglycemic Chinese patent medicine for obese T2DM， and to better guide the clinical promotion of this drug.

Objective:To investigate regulatory effect and mechanism of protein kinase C(PKC)δ/θ-dual functional oxidase 1(Duox1)-reactive oxygen species(ROS)signaling pathway on human airway mucin(MUC)5AC,to provide a new target for treatment of high secretion of airway mucus.Methods:Human airway epithelial cells 16HBE were pretreated with PKC and its subunit PKCδ/θ inhibitor,Duox1 inhibitor or free radical scavenger DMTU,respectively,and then human neutrophil elastase(HNE)stimulation to establish an in vitro airway inflammatory cell model.Generation level of ROS in each group of cells was determined by kit,mRNA levels of Duox1 and MUC5AC were detected by real-time fluorescent quantitative PCR,influence of interfering factors of each group of cells on Duox1 protein level was determined by Western blot,and protein expression of MUC5AC in each group of cells was detected by ELISA and immunofluorescence.Results:Compared with control group,ROS production in HNE group was increased significantly,expressions of Duox1 and MUC5AC mRNA and protein were also increased(P<0.05).After administration of Duox1 inhibitors,free radical scavengers or PKC inhibitors and PKCδ/θ inhibitors,ROS production was significantly inhibited,Duox1 and MUC5AC mRNA and protein expressions were decreased(P<0.05),while after giving PKCα/β,ROS generation,Duox1 and MUC5AC mRNA and pro-tein expressions were not significantly changed compared with HNE group(P>0.05).Conclusion:HNE can mediate high expression of MUC5AC through PKCδ/θ-Duox1-ROS,which plays an important role in development of high secretion of airway mucus in vitro cell model experiment.

Objective:Three-dimensional reconstruction and finite element analysis were used to investigate the outcomes of different fixation methods on horizontal osteotomy genioplasty and to identify the fixation method with best biomechanics.Methods:The skull spiral CT data of a patient with chin deformity were collected. The mandible three-dimensional model and internal fixation devices were reconstructed by Mimics, Geomagic Studio, Solidworks and Ansys Workbench software. The surgical osteotomy and four internal fixation methods (simple H-shaped titanium plate; 3 lag screws; H-shaped titanium plate combined with bilateral straight titanium plates; H-shaped titanium plate combined with bilateral lag screws) were simulated. The difference of deformation and stress distribution in each fixation under the same load were tested by finite element analysis.Results:The results of H-shaped titanium plate combined with bilateral lag screws fixation were the most balanced, the overall displacement was minimal (0.033 2 mm), the stress concentration of fixation material and free bone was 59.79 MPa and 4.69 MPa, respectively, and the regional stress was small[comparison of the equivalent stress of fixation material between groups, except that it was similar to the three lag screw fixation methods ( P>0.05), it was significantly smaller than the other two fixation methods ( P<0.01); comparison of the equivalent stress of free bone segment between groups, except that it was similar to the simple H-shaped titanium plate fixation method ( P>0.05), it was significantly smaller than the other two fixation methods ( P<0.05)]. Conclusion:The method of H-shaped titanium plate combined with bilateral lag screws fixation in horizontal osteotomy genioplasty has small deformation, which is not easy to cause deformation and fracture of fixation materials, and has superior mechanical properties. It is an appropriate method for clinical application.

Objective:To investigate the cosmetic effects of botulinum toxin type A(BTX-A) in the prevention and treatment of cicatrix on split-thickness skin graft donor site.Methods:A prospective randomized controlled study was commenced to recruit patients with functional burns and chronic wounds who were repaired with split-thickness skin graft in the Department of Burn and Plastic Surgery of Zhongshan People’s Hospital from September 2021 to September 2022. Ten days after wound healing in the donor area of the thigh (about 4 weeks after surgery), the patients were randomly divided into two groups: the experimental group and the control group. The experimental group was injected with BTX-A solution at the skin donor site by equal interval injection method 1 and 3 months after surgery, 0.1 ml (1 U) was injected at each point, and the interval of each point was 1 cm. The control group was injected with equal amount of normal saline at the skin donor area by equal spacing method 1 and 3 months after operation. The adverse reactions was observed. Follow-up was performed six months after treatment. Evaluation indicators included the doctor’s Vancouver scar scale (VSS) score (including color, vascular distribution, softness and thickness), the visual analogue scale (VAS) score of the patient’s scar pain and itching degree. Both VSS scores and VAS scores were in line with normal distribution, expressed as Mean±SD, and analyzed by independent sample t-test. Results:A total of 60 subjects were recruited, 30 in the experimental group and 30 in the control group. In the experimental group, there were 16 males and 14 females, aged (32.7±5.4) years (18-60 years). In the control group, there were 13 males and 17 females, aged (31.4±4.8) years (18-55 years). There were no significant differences in gender composition and age between the two groups (all P > 0.05). No serious adverse reactions occurred in the experimental group during and after treatment. At the follow-up six months after treatment, the total score of VSS in the experimental group [(3.57±0.60) points vs. (8.52±0.84) points] and the scores of scar color, vascular distribution, flexibility and thickness[(0.88±0.22) points vs. (2.30±0.52) points; (0.73±0.27) points vs. (2.16±0.40) points; (1.29±0.39) points vs. (2.49±0.39) points; (0.66±0.23) points vs. (1.56±0.34) points] were significantly lower than those in the control group, and the differences were statistically significant ( P < 0.01). The VAS score of scar pain and itching degree in the experimental group was also significantly lower than that in the control group [(1.06±0.34) points vs. (2.92±0.63) points], and the difference was statistically significant ( P < 0.01). Conclusion:BTX-A injection is safe and effective in preventing and treating cicatrix in the donor area of split-thickness skin graft, and also has a good effect on relieving the pain and itching symptoms of cicatrix patients.

Objective:To establish a nasal septum evaluation model before and after nasal septum deviation correction surgery with finite element analysis, to explore the effect of nasal septum cartilage stress reduction after nasal septum deviation correction surgery.Methods:In this study, a male adult patient treated in Zhujiang Hospital of Southern Medical University in January 2022 was selected for second-line nasal endoscopic tension-reduction septum correction. Three-dimensional reconstruction and finite element analysis of nasal septum cartilage were performed before surgery and 6 months after surgery, and DICOM format data of three-dimensional CT were collected. Mimics 21.0, Geomagic Studio 2014, and ANSYS Workbench 17.0 software were used to establish a three-dimensional finite element model of the nasal septum, and fixed constraints were set on the top and bottom of the nasal septum cartilage. A contact stress with a pressure of 10 6 Pa along the X axis was applied in the front plane of the cartilage, and then the finite element analysis of the total deformation displacement and von Mises stress (VMS) distribution of the nasal septum was performed. Results:(1) The maximum value of the total deformation displacement of the deviated nasal septum cartilage was located in the front of the cartilage, and the maximum value was 0.022 296 m before surgery and 0.108 810 m after surgery. (2) The stress distribution of deviated nasal septum cartilage was mainly concentrated in the anterior cartilage, in which VMS had a peak at the anterior end of the nasal septum, and the VMS peak was 725 000 Pa before surgery and 2 426 000 Pa after surgery. (3) The total deformation displacement and VMS value curves of the nasal septum showed a "waterfall" drop from the anterior end of the nasal septum to the posterior end. (4) The maximum distribution range of VMS in the nasal septum was 0.003 252 4 m 2 before surgery and 0.000 418 6 m 2 after surgery, indicating that the stress distribution range of the nasal septum after surgery was significantly reduced compared with that before surgery. Conclusion:The three-dimensional finite element model of nasal septum is established successfully through CT three-dimensional reconstruction and finite element analysis. By comparing the total deformation displacement and VMS curve before and after the operation, this model evidenced that the correction of nasal deviation has the effect of reducing the stress of nasal septum cartilage.

Allergic rhinitis(AR)is a typeⅠallergy of the nasal mucosa mediated by IgE.Allergen specific immunotherapy(AIT)can not only control nasal symptoms effectively and long term,but also change the natural course of disease.Due to the poor re-sponse of some patients to allergen specific immunotherapy and the long course of treatment,it is critical to identify biological indica-tors that can predict and evaluate the efficacy.This review reviews the progress in the study of biomarkers for the prediction and evalua-tion of the efficacy of sublingual immunotherapy(SLIT)for AR.The changes of various immune cells,immunoglobulin and cytokines are related to the efficacy of SLIT and are considered as candidate biomarkers,but their effectiveness and relevance remain to be fur-ther verified.