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 investigate the factors contributing to in-hospital mortality among elderly patients aged 80 and above with gastrointestinal bleeding(GIB).Additionally, it seeks to assess the predictive ability of the electronic frailty index(eFI)in determining the risk of in-hospital mortality in GIB patients.Methods:A retrospective analysis was performed among 624 patients aged 80 and above with GIB who were admitted to Beijing Hospital between July 2013 and September 2019.The patients were categorized into two groups based on their discharge outcomes: those who survived and those who did not.The eFI was developed using a cumulative deficit model utilizing data from the hospital's electronic medical records.The study examined the clinical features and risk factors associated with in-hospital mortality among these elderly patients.The effectiveness of eFI in predicting in-hospital mortality in elderly patients with gastrointestinal bleeding was evaluated by calculating the area under the curve(AUC)of the receiver operating characteristic(ROC)curve.Results:Among a total of 624 patients aged between 80 and 102 years, the average age was(83.0±6.4)years, with 339 being male.A majority of the patients, 581 cases(93.1%), had an eFI ≥ 0.15.A comparison between the survival group(380 cases)and the death group(244 cases)revealed that the latter had higher eFI values(0.39±0.09 vs.0.29±0.11, t=-11.452, P<0.001), along with higher rates of heart failure, chronic kidney disease, and malignant tumors, as well as lower body mass index, hemoglobin, albumin, and total cholesterol levels, and higher alanine aminotransferase and D-dimer levels(all P<0.05).Logistic regression analysis indicated that eFI( OR=2.322, 95% CI: 1.840-2.929, P<0.001), malignant tumor( OR=1.833, 95% CI: 1.141-2.860, P<0.001), and albumin<35 g/L( OR=1.826, 95% CI: 1.200-2.777, P<0.001)were independent risk factors for in-hospital death in elderly patients aged 80 and over with gastrointestinal bleeding.With every 0.1 increase in eFI, the risk of in-hospital death rose by 1.322 times.The AUC of eFI for predicting in-hospital mortality was 0.751(95% CI: 0.713-0.789, P<0.001).An eFI of ≥0.33 demonstrated a sensitivity of 77.9% and a specificity of 60.3% in predicting in-hospital mortality in elderly patients aged 80 and over with GIB. Conclusions:The eFI serves as an important independent risk factor for in-hospital mortality among patients aged 80 and above who experience GIB.It can effectively assess the prognosis of elderly individuals facing GIB.

Background::We previously reported that activation of the cell cycle in human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) enhances their remuscularization capacity after human cardiac muscle patch transplantation in infarcted mouse hearts. Herein, we sought to identify the effect of magnesium lithospermate B (MLB) on hiPSC-CMs during myocardial repair using a myocardial infarction (MI) mouse model.Methods::In C57BL/6 mice, MI was surgically induced by ligating the left anterior descending coronary artery. The mice were randomly divided into five groups ( n = 10 per group); a MI group (treated with phosphate-buffered saline only), a hiPSC-CMs group, a MLB group, a hiPSC-CMs + MLB group, and a Sham operation group. Cardiac function and MLB therapeutic efficacy were evaluated by echocardiography and histochemical staining 4 weeks after surgery. To identify the associated mechanism, nuclear factor (NF)-κB p65 and intercellular cell adhesion molecule-1 (ICAM1) signals, cell adhesion ability, generation of reactive oxygen species, and rates of apoptosis were detected in human umbilical vein endothelial cells (HUVECs) and hiPSC-CMs. Results::After 4 weeks of transplantation, the number of cells that engrafted in the hiPSC-CMs + MLB group was about five times higher than those in the hiPSC-CMs group. Additionally, MLB treatment significantly reduced tohoku hospital pediatrics-1 (THP-1) cell adhesion, ICAM1 expression, NF-κB nuclear translocation, reactive oxygen species production, NF-κB p65 phosphorylation, and cell apoptosis in HUVECs cultured under hypoxia. Similarly, treatment with MLB significantly inhibited the apoptosis of hiPSC-CMs via enhancing signal transducer and activator of transcription 3 (STAT3) phosphorylation and B-cell lymphoma-2 (BCL2) expression, promoting STAT3 nuclear translocation, and downregulating BCL2-Associated X, dual specificity phosphatase 2 (DUSP2), and cleaved-caspase-3 expression under hypoxia. Furthermore, MLB significantly suppressed the production of malondialdehyde and lactate dehydrogenase and the reduction in glutathione content induced by hypoxia in both HUVECs and hiPSC-CMs in vitro. Conclusions::MLB significantly enhanced the potential of hiPSC-CMs in repairing injured myocardium by improving endothelial cell function via the NF-κB/ICAM1 pathway and inhibiting hiPSC-CMs apoptosis via the DUSP2/STAT3 pathway.

Objective:To compare the clinical outcomes between pedicle screw internal fixation via the Wiltse approach and conservative treatment in young patients with thoracolumbar fracture with Thoracolumbar Injury Classification and Severity score (TLICS) ≤ 4 points.Methods:This retrospective study included 219 young patients with thoracolumbar fracture with TLICS score ≤ 4 points who had been treated from January 2014 to December 2018 at Department of Orthopaedics, The Second Hospital of Shanxi Medical University and obtained full follow-up. They were assigned into a surgery group of 126 patients subjected to pedicle screw internal fixation via the Wiltse approach and a conservative group of 93 patients subjected to conservative treatment. The surgery group included 65 males and 61 females, aged from 18 to 37 years, with a TLICS score of 1 point in 38 cases and of 2 to 4 points in 88 ones; the conservative group included 48 males and 45 females, aged from 19 to 38 years, with a TLICS score of 1 point in 29 cases and of 2 to 4 points in 64 ones. Patients in both groups underwent thoracolumbar X-ray, CT and MRI before treatment and regular thoracolumbar X-ray reexamination after treatment. Improvements in visual analog scale (VAS) for low back pain were compared between pre- and post-treatment. The 2 groups were compared in terms of VAS, anterior height of the injured vertebra and kyphosis cobb angle between pre-treatment, one month post-treatment and the last follow-up.Results:The 2 groups were comparable due to insignificant differences between them in the pre-treatment general data ( P>0.05). In the surgery group, patients were followed up for 24 to 72 months, the average VAS scores at one month post-treatment (2.5±1.2) and the last follow-up (2.3±0.8) were significantly improved compared to the pre-treatment value (6.8±2.1) ( P<0.05), and no serious surgical complications occurred. In the conservative group, patients were followed up for 30 to 65 months, the average VAS scores at one month post-treatment (3.9±1.9) and the last follow-up (3.5±0.9) were significantly improved compared to the pre-treatment value (6.2±2.0) ( P<0.05), and the rate of complications was 11.8% (11/93, including 3 cases of neural symptoms of the lower limb, 4 cases of bedsore and 4 cases of pulmonary infection). The VAS, anterior height of the injured vertebra and kyphosis cobb angle at one month post-treatment and the last follow-up in the surgery group were all significantly better than in the conservative group ( P<0.05). Conclusion:In young patients with thoracolumbar fracture with TLICS ≤ 4 points, pedicle screw internal fixation via the Wiltse approach can lead to better therapeutic outcomes than conservative treatment, especially in relief of postoperative low back pain.

Objective: To investigate the therapeutic effects and pharmacological mechanisms of Yishen Xingyang capsule (YXC) in oligoasthenospermia (OA) rats.Methods: Forty-eight male Sprague-Dawley rats were randomly divided into eight groups of six rats each:normal control (NC);model control (MC);three different positive drug (PD);and low-, medium-, and high-dose YXC groups. A rat model of OA was established by administering glucosides of Triptery-gium wilfordii Hook. F (GTW). After YXC administration, penile erectile function was observed. The epididymis, blood, and testes of the rats were harvested for analysis of sperm quality, sex hormone levels, mitochondrial membrane potential, and the transforming growth factor (TGF)-β1/Smad signaling pathway. Results: Compared with that in the MC group, penile erectile function in the YXC groups and three PD groups increased (all P<.01). Moreover, sperm quality in the YXC groups and three PD groups improved (all P < .001). The levels of testosterone, follicle stimulating hormone, and luteinizing hormone in the three PD and YXC groups increased (all P<.05). The mitochondrial membrane potential in the three PD and YXC groups significantly improved (all P<.001). Furthermore, the YXC and three PD groups showed decreased TGF-β1 expression (all P< .05) compared with the MC group. The high-dose YXC group and three PD groups improved Smad2 and Smad4 expression (all P<.05). Conclusion: YXC improved penile erectile function and sperm quality in OA rats, and the underlying mechanism included increase in sex hormones, inhibition of sperm apoptosis, and regulation of the TGF-β1/Smad signaling pathway. Meanwhile, this study provides a new effective drug option for the treat-ment of OA, which is beneficial to male reproductive health and social harmony.

0bjective: To explore the feasibility and clinical efficacy of cervical vertebral dome expansion laminoplasty.. Methods: Our hospital from February 2017 to 2018 Sep 16 cases with cervical spinal canal dome of cervical spinal stenosis angioplasty in treatment of cervical spinal cord due to the medical records of patients, including 14 males and 2 females; Aged 49- 76 years old, average age 57.3±1.7 years old. The course of disease was 7-48 months, with an average disease duration 17.75±1.90 months. Of the 16 patients, 5 had multi-segment cervical disc herniation and 11 had long segmental ossification of the posterior longitudinal ligament. All the 16 patients underwent cervical C_3-7 dome-type spinal canal enlargement; the position, stability and spinal cord compression of the internal fixation were evaluated according to the patient's imaging data (X-ray, CT and MRI). The neck and upper extremity pain was evaluated by visual analogue scale (VAS) before and after operation. The cervical spinal cord function was evaluated by the Japanese Orthopaedic Association (JOA) spinal cord function score, and the rate of improvement of neurological function was calculated. The Frankel grading was used to evaluate the neurological function of patients before and after surgery. Results: Cervical X-ray, CT and MRI were performed in all patients before and after surgery. Operation time 55-110 min, mean 65±12 min, bleeding 100-220 ml, mean 110±20 ml. The cross-sectional area of the spinal canal and the median sagittal diameter of the spinal canal were significantly increased compared with the preoperative. All patients were followed up for an average of 10.9±1.4 months (3-20 months). Imaging examination showed that 16 patients had no loosening and fracture after internal fixation, and no re-closure occurred. MRI T2WI images showed continuous recovery of cerebrospinal fluid signal in the spinal cord of C_3-7 range. The preoperative VAS score was 7.3±0.9 points, the average VAS score at the last follow-up was 1.6±0.4 points, the preoperative JOA score was 6.9±1.1 points, and the last follow-up JOA score was 13.4±1.3 points. The improvement rate was 87.23%±3.81%; Frankel grade D before surgery, and Frankel grade E after surgery. Conclusion Cervical spinal canal domed simple angioplasty operation, spinal canal full, satisfactory clinical efficacy, and can effectively reduce the incidence of related complications, it is a safe and feasible method for the treatment of cervical spinal stenosis.

Objective To prepare 3D printed porous tracheal graft fabricated by PCL and to select the appropriate pore size and surface modification techniques,in order to explore its effect on cell behavior.Methods The PCL porous tracheal graft was prepared by 3D printing technology and biomechanical properties of the graft were measured by means of longitudinal tension,radial compression and three-point bending test.The porous grafts were surface-modified through hydrolysis,amination and nanocrystallization treatment and then characterized by energy dispersive spectroscopy(EDS).The effect of different pore sizes and surface modifications on the cell proliferation behavior was evaluated by CCK-8 and scanning electron microscopy (SEM).Results The 3 D printed porous tracheal graft had similar morphology with the native tracheas(P ＞ 0.05) and better biomechanical properties(P ＜0.05).It was more suitable for cell adhesion and proliferation when the pore size is 200 μm (P ＜ 0.05).Compared to hydrolysis and amination,nanocrystallization treatment successfully improved the cytotropism of the 3D printed tracheal graft(P ＜ 0.05).Conclusion 3 D printed porous tracheal graft shows favorable biomechanical properties.The appropriate pore size of the 3D printed porous tracheal graft is 200 μm and the appropriate surface modification techniques is nanocrystallization.

Objective To investigate the effect of microRNA-1183 on proliferation and metastasis on gastric cancer cells and to explore the role of microRNA-1183 and CBL-B signaling pathways in this process. Methods MGC803 cells were transfected with a microRNA-1183 mimic. Real-time PCR detected the expression of microRNA-1183 in gastric cancer cell line MGC803. MTT detected the proliferative effect of microRNA-1183 on MGC803 gastric cancer cells. A Transwell assay detected the effect of microRNA-1183 on the metastasis of MGC803 gastric cancer cells. A dual luciferase reporter assay detected the binding ability between microRNA-1183 and CBL-B. The expression of the protein was tested by Western blotting. Results MTT assay results showed that microRNA-1183 promoted the proliferation of MGC803 cells. Transwell assay results revealed that microRNA-1183 promoted the metastasis of MGC803 cells. The results of BLAST contrast analysis show that CBL-B is one of the target genes of microRNA-1183. Western blotting analysis showed that the mimic microRNA-1183 inhibited the expression of CBL-B. A dual luciferase reporter assay showed that CBL-B was the target gene of microRNA-1183. A CBL-B knockdown promoted the proliferation and metastasis of MGC803 cells. microRNA-1183 promoted the proliferation and metastasis of MGC803 cells by inhibiting the expression of CBL-B. Conclusion microRNA-1183 can inhibit the proliferation and metastasis of gastric cancer cell lines by inhibiting the expression of CBL-B.

Objective To investigate the effect of Exosomes derived from lung cancer cells on the migration of secretory cells and homologous tumor cells and to explore the role of PI3K/Akt and SRC signaling pathways in this process.Methods Exosomes were isolated from the supematant post density gradient centrifugation of A549,lung cancer cells.Morphology of the Exosomes was studied using transmission electron microscopy.Protein expression was analyzed using Western blotting.Cell migration was analyzed by a transwell assay.Results The double-membrane-bound Exosomes appeared as discal-shaped structures,30-100 nm in diameter.Western blotting showed that CD9 was abundant in the Exosomes.The Exosomes promoted the migration of A549 cells and their homologous tumor cells,HCC827 in a dose-dependent manner,accompanied by the activation of Akt and SRC.Conclusion The Exosomes derived from A549 (lung cancer) cells promote the migration of the secreting cells and the homologous tumor cells.The mechanism may be correlated with the activation of Akt and SRC.

Objective:To establish a mouse model of gastric cancer by inoculating MKN45 cells into mice with normal immune function utilizing microcarrier technology. Methods:A total of 60 male C57BL/6 mice were randomly divided into three groups, namely, 2D, con-trol, and 3D groups, according to the coculture system of MKN45 and microcarrier. The mouse models of gastric carcinoma were estab-lished by hypodermic injection. The time of tumorigenesis, rate of tumor formation, and pathological features were observed in each group. Results:In the 3D group, the time of tumor formation was short, whereas the rate of tumor formation was high (80%). No de-tectable tumor formations were observed in the 2D and control groups. HE and immunohistochemical staining of the transplantation tumor model showed evident characteristics of human gastric cancer. Conclusion:A human gastric cancer model in normal immune mice was successfully established. The onset and development mechanism of gastric cancer could be more effectively investigated in mice with normal immune function through this model. Moreover, a more valuable and new animal model for the research and devel-opment of anticancer drug was established.