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.

BACKGROUND:Establishing a nomogram prediction model for postoperative pulmonary infection in hip fractures and taking early intervention measures is crucial for improving patients'quality of life and reducing medical costs. OBJECTIVE:To construct a nomogram risk prediction model of postoperative pulmonary infection in elderly patients with hip fracture,and provide theoretical basis for feasible prevention and early intervention. METHODS:Case data of 305 elderly patients with hip fractures who underwent surgical treatment at Wuxi Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine between January and October 2020(training set)were retrospectively analyzed.Using univariate and multivariate logistic regression analysis and Hosmer-Lemeshow goodness of fit test,receiver operating characteristic curve was utilized to analyze the diagnostic predictive efficacy of independent risk factors and joint models for postoperative pulmonary infections.Tools glmnet,pROC,and rms in R Studio software were applied to construct a nomogram model for predicting the risk of postoperative pulmonary infection in elderly patients with hip fractures,and calibration curves were further drawn to verify the predictive ability of the nomogram model.Receiver operating characteristic curves,calibration curves,and decision curves were analyzed for 133 elderly patients with hip fractures(validation set)receiving surgery at the same hospital from November 2022 to March 2023 to further predict the predictive ability of the nomogram model. RESULTS AND CONCLUSION:(1)The postoperative pulmonary infection rate in elderly patients with hip fractures in this group was 9.18%(28/305).(2)Single factor and multivariate analysis,as well as forest plots,showed that preoperative hospitalization days,leukocyte count,hypersensitive C-reactive protein,and serum sodium levels were independent risk factors(P<0.05).The Hosmer-Lemeshow goodness of fit test showed good fit(χ2=4.57,P=0.803).Receiver operating characteristic curve analysis was conducted on the independent risk factors and their joint models mentioned above,and the differentiation of each independent risk factor and joint model was good,with statistical significance(P<0.05).(3)The graphical calibration method,C-index,and decision curve were used to validate the nomogram prediction model.The predicted calibration curve was located between the standard curve and the acceptable line,and the predicted risk of the nomogram model was consistent with the actual risk.(4)The validation set used receiver operating characteristic curve,graphic calibration method,and decision curve to validate the prediction model.The results showed good consistency with clinical practice,indicating that the model had a good fit.The nomogram risk prediction model constructed for postoperative pulmonary infection in elderly patients with hip fractures has good predictive performance.The use of the nomogram risk prediction model can screen high-risk populations and provide a theoretical basis for early intervention.

Objective:To study the feasibility of a modified pancreatogastric anastomosis in laparoscopic duodenum preserving pancreatic head resection (LDPPHR).Methods:The clinical data of 25 patients with benign or low-grade malignant tumors of pancreatic head undergoing LDPPHR at the Second Hospital of Hebei Medical University from January 2019 to May 2023 were retrospectively analyzed, including 7 males and 18 females, aged (44.9±6.2) years old. According to the methods of pancreatic digestive reconstruction, patients were divided into the observation group ( n=10), who underwent the modified pancreatogastric anastomosis, and the control group ( n=15) who underwent conventional pancreaticojejunal anastomosis and jejuno-jejunal anastomosis. The general data, intraoperative pancreatic digestive reconstruction time, maximum levels of amylase in abdominal drainage within three days postoperatively, postoperative complications, and hospital stay were compared between the groups. Results:All procedures were performed successfully. The intraoperative pancreatic digestive reconstruction time was shorter in the observation group [(27.8±2.4) min vs. (45.8±3.6) min, P=0.010]. The intraoperative blood loss were comparable between the groups [(140.5±14.8) ml vs. (145.2±9.7) ml, P=0.843]. The maximum level of amylase in abdominal drainage within three days postoperatively was lower in the observation group [(809.1±185.5) U/L vs. (1 385.4±481.1) U/L, P=0.031]. No grade C pancreatic fistula or postoperative hemorrhage occurred in either group, and the incidence of grade B pancreatic fistula was lower in the observation group [20.0% (2/10) vs. 60.0% (9/15), P=0.048], with a shorter postoperative hospital stay [(7.9±1.3) d vs. (10.3±2.7) d, P=0.017]. No decrease of life quality or reoperation due to pancreatic fistula, hemorrhage or digestive tract malfunction occurred in either group within a median follow-up of 15.6 months. Conclusion:In LDPPHR, the modified pancreatogastric anastomosis could help shorten the pancreatic digestive reconstruction and lower the risk of postoperative pancreatic fistula.

The genus Tetrastigma belongs to the Vitaceae family and contains over 100 species. This paper reviewed folk uses, chemical constituents, pharmacological activities, and clinical applications of the medicinal plants in the genus Tetrastigma. In addition, the paper also discussed the current problems for the further studies. Up to now, more than 240 compounds were reported from the genus Tetrastigma, covering 74 flavonoids, 14 terpenoids, 19 steroids, 21 phenylpropanoids, 14 alkaloids and others constituents. Among them, flavonoids are the major and the characteristic chemical constituents in this genus. Modern pharmacological studies and clinical practice showed that the extracts and chemical constituents of Tetrastigma species possessed wide pharmacological activities including antitumor, antioxidative, hepatoprotective, antiviral, anti-inflammatory, and analgesic activities. The information summarized in this paper provides valuable clues for new drug discovery and an incentive to expand the research of genus Tetrastigma.

Objective:To investigate the differential expression of microRNA (miR)-214-3p in plasma exosomes in different types of uveal melanoma (UM) and evaluate whether miR-214-3p is a potential molecular biomarker for the diagnosis and prognosis of UM.Methods:Twenty-five UM in situ patients who received the enucleation of eyeball were enrolled at Beijing Tongren Hospital from December 2015 to October 2019, including 10 with epithelioid cell melanoma and 10 with spindle cell melanoma as well as 5 metastatic UM patients (1 with spindle cell-like melanoma and 4 with epithelioid cell-like melanoma) and 10 healthy subjects were enrolled during the same period.Blood sample was collected from all the subjects for the isolation of plasma exosomes.The morphology of exosomes was examined under the electron microscope.The exsomal marker proteins were identified by Western blot.The expression level of miR-214-3p in plasma exosomes was detected by real-time fluorescence quantitative PCR.The differential expression of miR-214-3p among different types of UM patients and healthy controls was compared.The diagnostic and classification performance of exosomal miR-214-3p was evaluated using receiver operating characteristic curve.Written informed consent was obtained from each subject prior to entering the study cohort.This study protocol was approved by an Ethics Committee of Capital Medical Univeristy (No.TRECKY2018-056).Results:The isolated exosomes were hemispherical in shape with a concavity on one side.The diameter of the exosomes was about 100 nm and the particle diameter of vesicles from samples was (82.0±2.7) nm.TSG101 protein was detectable and Calnexin protein was not found in the exosomes.The relative expression levels of plasma exosomal miR-214-3p in healthy control group, in situ UM group and metastatic UM group were 0.86(0.57, 1.49), 0.24(0.10, 0.67), and 0.43(0.23, 0.56), respectively.The miR-214-3p relative expression level in plasma exosomes of in situ UM patients and metastatic UM patients was significantly lower than that of healthy controls, and the differences were statistically significant ( Z=2.62, P<0.01; Z=2.08, P<0.05). The relative expression levels of exosomal miR-214-3p in spindle cell-like UM group and epithelioid cell-like UM group were 0.11(0.07, 0.64) and 0.46(0.14, 0.91), respectively, and no significant difference was found in the expression level of plasma exosomal miR-214-3p among different types of UM (all at P>0.05). The area under the curve of plasma exosomal miR-214-3p for UM diagnosis was 0.795. Conclusions:Plasma exosomal miR-214-3p level is significantly reduced in both in situ UM patients and metastatic UM patients.Plasma exosomal miR-214-3p is a new potential diagnostic biomarker for UM, but the exosomal miR-214-3p appears to not be able to distinguish the types of UM.

Objective:To investigate the clinical safety and efficacy of tirofiban in the treatment of hemiplegic stroke warning syndrome.Methods:Patients with hemiplegic stroke warning syndrome admitted to Jining First People's Hospital without receiving intravenous thrombolysis from January 2018 to May 2020 were enrolled retrospectively. Some patients were given tirofiban intravenous infusion for at least 24 h in acute phase, then received oral antiplatelet therapy (tirofiban group); some only received aspirin+ clopidogrel dual antiplatelet therapy (control group). The primary endpoint was muscle strength at the paralytic side and National Institutes of Health Stroke Scale (NIHSS) score at day 7 after onset. The secondary endpoint was the modified Rankin Scale (mRS) score at 3 months after onset, and ≤2 was defined as good clinical outcome. The safety endpoint was the bleeding events during treatment. Multivariate logistic regression analysis was used to determine the independent influencing factors of clinical outcome. Results:A total of 30 patients with hemiplegic stroke warning syndrome were enrolled, including 19 (63.3%) in the tirofiban group and 11 (36.7%) in the control group. There was no significant difference in baseline clinical data between the two groups, and no drug-related bleeding complications occurred during treatment. The muscle strength at paralytic side and NIHSS score at day 7 after onset, NIHSS score at discharge and good clinical outcome rate at 3 months in the tirofiban group were significantly better than those in the control group, and the differences were statistically significant (all P<0.05). Multivariate logistic regression analysis showed that tirofiban was an independent protective factor for good outcome after adjusting the NIHSS score at the beginning of treatment (odds ratio 0.040, 95% confidence interval 0.040-0.449; P=0.009). Conclusions:Tirofiban is safe and effective in the treatment of patients with hemiplegic stroke warning syndrome in acute phase. It can effectively block the progress of the disease, improve the outcome of patients, and will not increase the risk of bleeding.

The mammalian central nervous system (CNS) is considered an immune privileged system as it is separated from the periphery by the blood brain barrier (BBB). Yet, immune functions have been postulated to heavily influence the functional state of the CNS, especially after injury or during neurodegeneration. There is controversy regarding whether adaptive immune responses are beneficial or detrimental to CNS injury repair. In this study, we utilized immunocompromised SCID mice and subjected them to spinal cord injury (SCI). We analyzed motor function, electrophysiology, histochemistry, and performed unbiased RNA-sequencing. SCID mice displayed improved CNS functional recovery compared to WT mice after SCI. Weighted gene-coexpression network analysis (WGCNA) of spinal cord transcriptomes revealed that SCID mice had reduced expression of immune function-related genes and heightened expression of neural transmission-related genes after SCI, which was confirmed by immunohistochemical analysis and was consistent with better functional recovery. Transcriptomic analyses also indicated heightened expression of neurotransmission-related genes before injury in SCID mice, suggesting that a steady state of immune-deficiency potentially led to CNS hyper-connectivity. Consequently, SCID mice without injury demonstrated worse performance in Morris water maze test. Taken together, not only reduced inflammation after injury but also dampened steady-state immune function without injury heightened the neurotransmission program, resulting in better or worse behavioral outcomes respectively. This study revealed the intricate relationship between immune and nervous systems, raising the possibility for therapeutic manipulation of neural function via immune modulation.

Objective To investigate and analyze the clinical effect of laryngeal mask airway inhalation anesthe-sia with sevoflurane on pediatric colic surgery.Methods From July 2014 to November 2016,92 cases with pediatric hernia in Jiaxing Hospital of Zhejiang Corps of Armed Police Force were randomly divided into study group and control group according to the digital table ,with 46 patients in each group.All patients were selected according to the specific circumstances given hernia surgery ,sevoflurane inhalation anesthesia with laryngeal mask airway in the study group in the operation,the control group was given tracheal intubation anesthesia.The changes of vital signs ,the anesthesia effect and treatment satisfaction etc.were observed and analyzed.Results All the patients were treated by surgery , the heart rate (HR) and mean arterial pressure(MAP) changes in the study group were significantly better than those in the control group(all P＜0.05).The eye opening time and awake time in the study group were (7.28 ±1.07)min, (12.96 ±1.38 ) min, respectively, which were significantly better than those in the control group [ ( 13.02 ± 1.75)min,(21.13 ±2.82)min] (t＝18.979,P＝0.001;t＝17.649,P＝0.001).The total satisfaction rate of the study group was 97.83%,which was significantly higher than 82.61%of the control group (χ2＝13.126＝0.002). Conclusion The application of laryngeal surgery in children with hernia in mask ventilation of sevoflurane anesthe -sia,has good anesthetic effect ,it has less influence on hemodynamics in children ,and rapid postoperative recovery.

Objective To investigate the efficacy and accuracy of 3D printing individualized navigation templates used in shoulder hemi-arthroplasty (HA).Methods We collected 9 adult cadaveric specimens of 18 shoulders with no obvious deformities or defects.After CT scanning was conducted on bilateral shoulders,the raw CT data in DICOM format were imported into software Mimics 10.01 and Imageware 12.0 to reconstruct 3D shoulder models.The retroversion angles and heights of bilateral humeral heads were measured to design individualized navigational templates using software Geomagic Studio 12.0.Rapid prototyping (RP) technique was used to manufacture the individualized navigation templates which were then used to assist shoulder HA in cadaveric specimens.CT scanning was performed postoperatively to measure the retroversion angle and height of the prosthesis implanted for comparison with the preoperative measurements.Results Before shoulder replacement in the 9 cadaveric specimens,respectively,the heights of left and right humeral heads averaged 300.0 ± 11.6 mm and 300.0 ± 11.4 mm,and the retroversion angles 15.6°±4.9° and 15.9°± 4.9°.After the replacement,the heights of left and right humeral heads averaged 300.0 ± 11.3 mm and 300.0 ± 11.7 mm,respectively,and the retroversion angles 15.7° ± 4.9° and 15.8° ± 5.2°,respectively.There were no significant differences between the left and right sides in the pre-or post-operative heights or retroversion angles of the humeral head (P ＞ 0.05).There were no significant differences either between preand post-operation in the heights or retroversion angles of the left and right humeral heads (P ＞ 0.05).The mean errors for the preoperative and postoperative heights and retroversion angles of the humeral heads were 1.44 ± 0.96 mm and 0.25° ± 0.15°,respectively.Conclusion Individualized navigation templates 3D printed can be used to assist effective and accurate shoulder HA because they are designed and manufactured according to accurate measurements of retroversion angles and heights of the humeral heads which are acquired by computer preoperatively.

Objective To study the clinical application of midazolam combined with remifentanil in spinal anesthesia.Methods80 cases of vertebral fractures in Jiaxing hospital, Zhejiang armed police corps from February 2014 to December 2016 were selected as the research object in this study.They were randomly divided into two groups: control group and experimental group, each group had a total of 40 cases.Patients in the control group were treated with midazolam anesthesia, the experimental group was treated with midazolam combined with remifentanil anesthesia.Comparative analysis of the experimental group and control group of patients with vital signs and anesthesia recovery time and anesthetic effect indicators.ResultsAfter the corresponding anesthesia, the respiratory frequency and heart rate of the experimental group were significantly higher than those of the control group, with statistical difference (P<0.05).There was no significant difference between the control group and the experimental group, no statistical significance.The recovery time, the time of open eyes and extubation time of the experimental group were significantly lower than those of the control group (P<0.05).The excellent rate of anesthesia in the experimental group was 95.0%, and the excellent rate of anesthesia in the control group was 75.0%.As a result, the excellent rate of anesthesia in the control group was significantly lower than that in the experimental group, the OAAS score of the experimental group was significantly higher than that of the control group, with statistical difference (P<0.05).ConclusionThe application of midazolam combined with remifentanil in anesthesia of spinal fracture surgery, anesthesia effect is good, the patient's recovery time is faster, with the clinical significance of further promotion and application.