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 clinical application effects of a domestic bone-level implant system for restoring single tooth loss, and provide clinical evidence for the promotion and application of domestic implants.Methods:A prospective, multicenter clinical trial was conducted from April 2018 to January 2020 in three institutions: Department of Oral Implantology, School & Hospital of Stomatology, Wuhan University, Department of Stomatology, The First Affiliated Hospital of Zhejiang University School of Medicine, and Department of Stomatology, The Third Hospital of Hebei Medical University. The trial planned to include 100 patients for single tooth implantation and restoration, followed up for 1 year, to evaluate the implantation success rate and other related outcomes.Results:This study screened a total of 142 patients and ultimately included 100, comprising 43 males and 57 females with age of (47.0±12.2) years. Ninety-eight out of 100 patients completed a one-year follow-up (98.0%), while 2 patients terminated the trial early due to implant loosening (2.0%). After a one-year follow-up, the implants of the 98 patients were all functioning successfully, with a success rate of 98.0% (98/100). The patients were satisfied with the overall restoration effect.Conclusions:This study indicates that the domestic bone-level implant system has achieved favorable short-term clinical outcomes for single-tooth implantation and restoration.

Objective:To compare the outcomes of robot-assisted laparoscopic partial nephrectomy (RAPN) and laparoscopic partial nephrectomy (LPN) in the treatment of tumors in isolated kidney, and analyze the factors influencing postoperative renal function and long-term survival in patients.Methods:A retrospective analysis was conducted on clinical data of 67 patients with tumors in isolated kidney who underwent surgery at the Chinese PLA General Hospital from November 2010 to January 2022. There were 48 males and 19 females, with an average age of (58.6±10.1) years old. The patients were divided into RAPN group (43 cases) and LPN group (24 cases) based on the surgical approach. The RAPN group had a higher R.E.N.A.L. score than the LPN group [(8.7±1.5) vs. (7.9±1.7), P=0.042]. There were no statistically significant differences between the two groups in terms of age [(57.4±10.2) years old vs. (60.9±9.8) years old, P=0.185], body mass index (BMI) [(25.7±3.5) kg/m 2 vs. (25.1±3.6) kg/m 2, P=0.518], and preoperative serum creatinine [(102.9±31.6) μmol/L vs. (102.3±22.4) μmol/L, P=0.930]. Twelve cases underwent hypothermic treatment during surgery, with 9 cases(20.9%) in the RAPN group and 3 cases(12.5%) in the LPN group( P=0.596). Surgical time, intraoperative warm ischemia time, intraoperative blood loss, postoperative fasting time, perioperative complication rate, postoperative serum creatinine, and other indicators were compared between the two groups. Multiple linear regression analysis was used to identify factors affecting postoperative serum creatinine. Kaplan-Meier curves were employed to analyze patient prognosis, and log-rank tests were performed to compare the differences between the two groups. Multiple Cox regression analysis was used to identify factors influencing patient prognosis. Results:All surgeries were completed successfully with negative pathological margins. There were no statistically significant differences between the RAPN and LPN groups in terms of surgical time [(136.6±47.6) min vs. (125.3±34.4) min, P=0.311], intraoperative ischemia time [23.0 (16.0, 30.0) min vs. 19.0 (13.5, 27.5) min, P =0.260], intraoperative blood loss [50.0 (50.0, 100.0) ml vs. 50.0 (22.5, 100.0) ml, P=0.247], postoperative hospital stay [(6.6±3.5) days vs. (7.7±4.2) days, P=0.244], time to drain removal [4(3, 5) days vs. 5(3, 6) days, P =0.175], postoperative fasting time [(2.1±0.7) days vs. (2.2±1.0) days, P=0.729], perioperative complication rate [18.6% (8/43) vs. 16.7% (4/24), P=1.000], postoperative serum creatinine [145.2 (128.3, 191.3) μmol/L vs. 157.8 (136.2, 196.3) μmol/L, P =0.229], and pathological staging [T 1a/T 1b/T 2a/T 3a/T 4 stage: 32/7/1/3/0 case vs. 17/5/0/1/1 case, P=0.804]. Kaplan-Meier survival curves showed that the total survival rates at 1, 3, and 5 years after surgery were 94.7%, 84.9%, and 84.9% for the RAPN group, and 100.0%, 95.5%, and 95.5% for the LPN group, with no statistically significant difference in the log-rank test ( P=0.116). Excluding 10 patients with preoperative tumor metastasis (7 in the RAPN group and 3 in the LPN group), the progression-free survival rates at 1, 3, and 5 years after surgery were 84.8%, 81.1%, and 81.1% for the RAPN group, and 100.0%, 95.0%, and 90.0% for the LPN group, with no statistically significant difference in the log-rank test ( P =0.142). Multiple linear regression analysis showed that the use of hypothermic treatment during surgery significantly reduced postoperative serum creatinine ( B=-72.191, P=0.048). Multiple Cox regression analysis revealed that BMI ( HR=0.743, P=0.044), pathological T stage ( HR=4.235, P=0.018), and preoperative metastasis ( HR=18.829, P=0.035) were independent factors affecting patient overall survival time. A smaller BMI, higher pathological stage, and preoperative metastasis were associated with poorer prognosis. Conclusions:Despite the higher R. E.N.A.L. score and greater surgical difficulty in the RAPN group, RAPN achieved similar perioperative and prognostic results as the LPN, indicating RAPN advantages in treating tumors in isolated kidney. Appropriate intraoperative hypothermic treatment can better protect postoperative renal function. BMI, pathological T stage, and preoperative metastasis are independent factors affecting overall survival time.

Objective:To investigate the incidence of postoperative complications in Chinese patients with gastric or colorectal cancer, and to evaluate the risk factors for postoperative complications.Methods:This was a national, multicenter, prospective, registry-based, cohort study of data obtained from the database of the Prevalence of Abdominal Complications After Gastro- enterological Surgery (PACAGE) study sponsored by the China Gastrointestinal Cancer Surgical Union. The PACAGE database prospectively collected general demographic characteristics, protocols for perioperative treatment, and variables associated with postoperative complications in patients treated for gastric or colorectal cancer in 20 medical centers from December 2018 to December 2020. The patients were grouped according to the presence or absence of postoperative complications. Postoperative complications were categorized and graded in accordance with the expert consensus on postoperative complications in gastrointestinal oncology surgery and Clavien-Dindo grading criteria. The incidence of postoperative complications of different grades are presented as bar charts. Independent risk factors for occurrence of postoperative complications were identified by multifactorial unconditional logistic regression.Results:The study cohort comprised 3926 patients with gastric or colorectal cancer, 657 (16.7%) of whom had a total of 876 postoperative complications. Serious complications (Grade III and above) occurred in 4.0% of patients (156/3926). The rate of Grade V complications was 0.2% (7/3926). The cohort included 2271 patients with gastric cancer with a postoperative complication rate of 18.1% (412/2271) and serious complication rate of 4.7% (106/2271); and 1655 with colorectal cancer, with a postoperative complication rate of 14.8% (245/1655) and serious complication rate of 3.0% (50/1655). The incidences of anastomotic leakage in patients with gastric and colorectal cancer were 3.3% (74/2271) and 3.4% (56/1655), respectively. Abdominal infection was the most frequently occurring complication, accounting for 28.7% (164/572) and 39.5% (120/304) of postoperative complications in patients with gastric and colorectal cancer, respectively. The most frequently occurring grade of postoperative complication was Grade II, accounting for 65.4% (374/572) and 56.6% (172/304) of complications in patients with gastric and colorectal cancers, respectively. Multifactorial analysis identified (1) the following independent risk factors for postoperative complications in patients in the gastric cancer group: preoperative comorbidities (OR=2.54, 95%CI: 1.51-4.28, P<0.001), neoadjuvant therapy (OR=1.42, 95%CI:1.06-1.89, P=0.020), high American Society of Anesthesiologists (ASA) scores (ASA score 2 points:OR=1.60, 95% CI: 1.23-2.07, P<0.001, ASA score ≥3 points:OR=0.43, 95% CI: 0.25-0.73, P=0.002), operative time >180 minutes (OR=1.81, 95% CI: 1.42-2.31, P<0.001), intraoperative bleeding >50 mL (OR=1.29,95%CI: 1.01-1.63, P=0.038), and distal gastrectomy compared with total gastrectomy (OR=0.65,95%CI: 0.51-0.83, P<0.001); and (2) the following independent risk factors for postoperative complications in patients in the colorectal cancer group: female (OR=0.60, 95%CI: 0.44-0.80, P<0.001), preoperative comorbidities (OR=2.73, 95%CI: 1.25-5.99, P=0.030), neoadjuvant therapy (OR=1.83, 95%CI:1.23-2.72, P=0.008), laparoscopic surgery (OR=0.47, 95%CI: 0.30-0.72, P=0.022), and abdominoperineal resection compared with low anterior resection (OR=2.74, 95%CI: 1.71-4.41, P<0.001). Conclusion:Postoperative complications associated with various types of infection were the most frequent complications in patients with gastric or colorectal cancer. Although the risk factors for postoperative complications differed between patients with gastric cancer and those with colorectal cancer, the presence of preoperative comorbidities, administration of neoadjuvant therapy, and extent of surgical resection, were the commonest factors associated with postoperative complications in patients of both categories.

Objective:To observe and evaluate the clinical efficacy and safety of albumin-bound paclitaxel as first-line treatment for patients with advanced pancreatic cancer in China, and to explore the prognosis-related molecules in pancreatic cancer based on next-generation sequencing (NGS) of tumor tissues.Methods:From December 2018 to December 2020, patients with locally advanced or metastatic pancreatic cancer were recruited to accept albumin-bound paclitaxel as first-line treatment in the oncology departments of 24 hospitals in East China. The primary endpoints were overall survival (OS) and treatment related adverse events, and the secondary endpoint was progression-free survival (PFS). Adverse effects were graded using Common Terminology Criteria for Adverse Events 5.0 (CTCAE 5.0). NGS sequencing on the primary or metastatic tissue samples of pancreatic cancer obtained through surgical resection or biopsy was performed.Results:This study recruited 229 patients, including 70 patients with locally advanced pancreatic cancer (LAPC) and 159 patients with metastatic pancreatic cancer (mPC). The disease control rate was 79.9% and the objective response rate is 36.3%.The common adverse effects during treatment were anaemia (159 cases), leucopenia (170 cases), neutropenia (169 cases), increased aminotransferases (110 cases), and thrombocytopenia (95 cases), and the incidence of grade 3-4 neutropenia is 12.2% (28/229). The median follow-up time was 21.2 months (95% CI: 18.5-23.1 months). The median PFS (mPFS) was 5.3 months (95% CI: 4.37-4.07 months) and the median OS (mOS) was 11.2 months (95% CI: 9.5-12.9 months). The mPFS of patients with LAPC was 7.4 months (95% CI: 6.6-11.2 months), and their mOS was 15.5 months (95% CI: 12.6-NA months). The mPFS of patients with mPC was 3.9 months (95% CI: 3.4-5.1 months), and their mOS was 9.3 months (95% CI: 8.0-10.8 months). Multivariate Cox regression analysis showed that clinical stage ( HR=1.47, 95% CI: 1.06-2.04), primary tumor site ( HR=0.64, 95% CI: 0.48-0.86), Eastern Cooperative Oncology Group Performance Status (ECOG PS) score ( HR=2.66, 95% CI: 1.53-4.65), and whether to combine radiotherapy ( HR=0.65, 95% CI: 0.42-1.00) were independent influencing factors for the PFS of these patients. The primary tumor site ( HR=0.68, 95% CI: 0.48-0.95), ECOG score ( HR=5.82, 95% CI: 3.14-10.82), and whether to combine radiotherapy ( HR=0.58, 95% CI: 0.35-0.96) were independent influencing factors of the OS of these patients. The most frequent gene mutations in these advanced stage pancreatic patients were KRAS (89.66%), TP53 (77.01%), CDKN2A (32.18%), and SMAD4 (21.84%) by NGS of tumor tissues from 87 pancreatic cancer patients with sufficient specimens. Further analysis revealed that mutations in CDKN2B, PTEN, FGF6, and RBBP8 genes were significantly associated with an increased risk of death ( P＜0.05). Conclusion:Albumin-bound paclitaxel as first-line treatment demonstrated feasible anti-tumor efficacy and manageable safety for patients with advanced pancreatic cancer in China.

Ankylosing spondylitis (AS) combined with lower cervical fracture is often categorized into unstable fracture, with a high incidence of neurological injury and a high rate of disability and morbidity. As factors such as shoulder occlusion may affect the accuracy of X-ray imaging diagnosis, it is often easily misdiagnosed at the primary diagnosis. Non-operative treatment has complications such as bone nonunion and the possibility of secondary neurological damage, while the timing, access and choice of surgical treatment are still controversial. Currently, there are no clinical practice guidelines for the treatment of AS combined with lower cervical fracture with or without dislocation. To this end, the Spinal Trauma Group of Orthopedics Branch of Chinese Medical Doctor Association organized experts to formulate Clinical guidelines for the treatment of ankylosing spondylitis combined with lower cervical fracture in adults ( version 2024) in accordance with the principles of evidence-based medicine, scientificity and practicality, in which 11 recommendations were put forward in terms of the diagnosis, imaging evaluation, typing and treatment, etc, to provide guidance for the diagnosis and treatment of AS combined with lower cervical fracture.

Objective:To investigate the efficacy and influencing factors of selective ligation of the middle meningeal artery after drilling and drainage surgery for chronic subdural hematoma.Methods:The clinical data of patients with chronic subdural hemorrhage who underwent routine drilling and drainage surgery in Beijing Geriatric Hospital from October 2018 to January 2021 were taken as the control group; The clinical data of patients with chronic subdural hemorrhage who underwent routine drilling and drainage surgery and ligation of the middle meningeal artery in the same hospital from February 2021 to January 2023 was taken as the observation group.Results:Comparison of CSDH Markwalder grading scale and Glasgow coma scale (MGS-GCS) neurosurgical dysfunction grading before and after surgery between the control group and the observation group showed statistically significant differences ( χ2 = 34.56 and 63.39, P<0.01). There was statistically significant difference in the grading of postoperative MGS-GCS neurosurgical dysfunction between the control group and the observation group ( χ2 = 4.34, P = 0.037); the recurrence rate of the observation group was lower than that of the control group: 10.6% (5/47) vs. 28.0% (14/50), with statistical differences ( χ2 = 4.63, P = 0.031). The factors that reduce recurrence in the middle meningeal artery ligation group included Alzheimer disease coronary heart disease, males, oral antipsychotic and oral anticoagulant ( χ2 = 15.25, 4.36, 6.09, 12.63 and 8.68; P<0.05). Conclusions:Preliminary exploration has found that selective ligation of the middle meningeal artery during the drilling and drainage surgery for chronic subdural hematoma can reduce postoperative recurrence, especially suitable for patients with Alzheimer disease, coronary heart disease, males, oral antipsychotic and oral anticoagulant. After preoperative CT skull reconstruction and pruning to obtain the sulcus of the middle meningeal artery, the body surface localization of the ligation of the middle meningeal artery can be obtained. This surgery is safe and there is no additional damage to the cranial nerve. The efficacy of this surgical technique is expected to be further validated through large-scale, multicenter, randomized controlled studies.

Due to the non-targeted release and low solubility of anti-gastric cancer agent,apatinib(Apa),a first-line drug with long-term usage in a high dosage often induces multi-drug resistance and causes serious side effects.In order to avoid these drawbacks,lipid-film-coated Prussian blue nanoparticles(PB NPs)with hyaluronan(HA)modification was used for Apa loading to improve its solubility and targeting ability.Furthermore,anti-tumor compound of gamabufotalin(CS-6)was selected as a partner of Apa with reducing dosage for combinational gastric therapy.Thus,HA-Apa-Lip@PB-CS-6 NPs were constructed to synchro-nously transport the two drugs into tumor tissue.In vitro assay indicated that HA-Apa-Lip@PB-CS-6 NPs can synergistically inhibit proliferation and invasion/metastasis of BGC-823 cells via downregulating vascular endothelial growth factor receptor(VEGFR)and matrix metalloproteinase-9(MMP-9).In vivo assay demonstrated strongest anti-tumor growth and liver metastasis of HA-Apa-Lip@PB-CS-6 NPs adminis-tration in BGC-823 cells-bearing mice compared with other groups due to the excellent penetration in tumor tissues and outstanding synergistic effects.In summary,we have successfully developed a new nanocomplexes for synchronous Apa/CS-6 delivery and synergistic gastric cancer(GC)therapy.

Objective To investigate the clinical significance of the expression of antioxidant 1 copper chaperone protein(ATOX1)in hepatocellular carcinoma(HCC)and its relationship with tumor proliferation,migration and invasion.Methods The expression of ATOX1 mRNA in HCC cancer tissue and normal liver tissue was analyzed using the Human Genome Atlas database.Immunohistochemical experiment was used to detect the expression of ATOX1 in 15 cases of HCC cancer tissue and adjacent tissue.Human HCC cell lines Hep3B and HepG2 were divided into the control group(NC),the ATOX1 knockdown group 1(si-ATOX1#1)and the ATOX1 knockdown group 2(si-ATOX1#2).The effects of ATOX1 knockdown on the malignant biological behavior of HCC cells were observed through CCK-8 cell proliferation experiment,scratch experiment and Transwell invasion experiments.A nude mouse xenograft tumor model was constructed to analyze the effect of ATOX1 knockdown on the quality and volume of transplanted tumors.Western blot assay was used to detect the relationship between ATOX1 and JAK2/STAT3 pathway protein expression.Results Bioinformatics analysis showed that expression of ATOX1 mRNA in HCC cancer tissue was higher than that in adjacent normal tissue(P＜0.05).The immunohistochemical staining results showed that the positive rate of ATOX1 protein was higher in HCC cancer tissue than that in adjacent tissue(93.33%vs.13.33%,P＜0.01).In vitro experimental results showed that siRNA knockdown of ATOX1 protein expression in Hep3B and HepG2 cells significantly reduced the proliferation,migration and invasion abilities of cancer cells(P＜0.05).In vivo experiments in mice showed that the volume and weight of subcutaneous xenograft tumors were significantly smaller in the sh-ATOX1 group than those in the sh-con group(P＜0.05).The expression levels of JAK2/STAT3 pathway-related proteins p-JAK2,p-STAT3,CyclinD1 and MMP2 were significantly lower in the subcutaneous transplanted tumor tissue of the sh-ATOX1 group than that of the sh-con group(P＜0.05).Conclusion ATOX1 can promote the proliferation,migration and invasion of HCC through JAK2/STAT3 pathway,which can potentially become a potential tumor marker and therapeutic target.

Objective To investigate the role of necroptosis key genes in spinal cord injury using bioinformatics methods to provide new targets for the diagnosis and treatment of spinal cord injury.Methods The peripheral blood transcriptome data of spinal cord injury samples(n=38)and healthy control samples(n=10)were obtained from GSE151371 data set in Gene Expression Omnibus(GEO)database.R software was used to identify differentially expressed genes and perform functional enrichment analysis.Machine learning algorithms(random forest and LASSO)and protein-protein interaction(PPI)networks are used to screen for necroptosis key genes and construct a diagnostic nomogram for spinal cord injury.Establish a rat spinal cord injury model to further verify the expression of necroptosis key genes by Western blotting and immunofluorescence staining.Results A total of 2050 differentially expressed genes were identified in the two groups.KEGG pathway enrichment analysis showed that the differentially expressed genes were involved in the nucleotide-binding oligomerization domain(NOD)-like receptor signaling pathway,hematopoietic cell lineage,and necroptosis;GO enrichment analysis showed that the differentially expressed genes were involved in the activation of leukocytes,tertiary granulation,and regulation of the defense response,and so on.Intersection analysis screened 15 necroptosis differentially expressed genes.KEGG pathway enrichment analysis showed that necroptosis differentially expressed genes were involved in necroptosis,influenza,and NOD-like receptor signaling pathways;GO enrichment analysis showed that necroptosis differentially expressed genes were significantly enriched in the cellular response to cytokine stimulation,cytokine-mediated signaling pathways,and response to cytokines.Integration of two machine learning algorithms and PPI analysis further screened two necroptosis key genes(IL1B and PLA2G4A).The nomogram established using IL1B and PLA2G4A can be used for early prediction of the occurrence of spinal cord injury.The validation results of the rat spinal cord injury model showed that the protein expression of IL-1β and PLA2G4A in the spinal cord injury group were significantly higher than those in the sham group(P<0.05).Conclusions IL1B and PLA2G4A as key genes of necroptosis involved in the development of spinal cord injury,can be used to predict the development of spinal cord injury with the promise of being new targets for the prevention and treatment of spinal cord injury.