Establishment of a hydrodynamic delivery system in ducks.

Chronic hepatitis B virus (HBV) poses a significant global health challenge as it can lead to acute or chronic liver disease and hepatocellular carcinoma (HCC). To establish a safety experimental model, a homolog of HBV-duck HBV (DHBV) is often used for HBV research. Hydrodynamic-based gene delivery (HGD) is an efficient method to introduce exogenous genes into the liver, making it suitable for basic research. In this study, a duck HGD system was first constructed by injecting the reporter plasmid pLIVE-SEAP via the ankle vein. The highest expression of SEAP occurred when ducks were injected with 5 µg/mL plasmid pLIVE-SEAP in 10% bodyweight volume of physiological saline for 6 s. To verify the distribution and expression of exogenous genes in multiple tissues, the relative level of foreign gene DNA and ß-galactosidase staining of LacZ were evaluated, which showed the plasmids and their products were located mainly in the liver. Additionally, ß-galactosidase staining and fluorescence imaging indicated the delivered exogenous genes could be expressed in a short time. Further, the application of the duck HGD model on DHBV treatment was investigated by transferring representative anti-HBV genes IFNα and IFNγ into DHBV-infected ducks. Delivery of plasmids expressing IFNα and IFNγ inhibited DHBV infection and we established a novel efficient HGD method in ducks, which could be useful for drug screening of new genes, mRNAs and proteins for anti-HBV treatment.

Pulley Effect in the Capture of DNA Translocation through Solid-State Nanopores.

Nanopores are powerful single-molecule sensors for analyzing biomolecules such as DNA and proteins. Understanding the dynamics of DNA capture and translocation through nanopores is essential for optimizing their performance. In this study, we examine the effects of applied voltage and pore diameter on current blockage, translocation time, collision, and capture location by translocating λ-DNA through 5.7 and 16 nm solid-state nanopores. Ionic current changes are used to infer DNA conformations during translocation. We find that translocation time increases with pore diameter, which can be attributed to the decrease of the stall force. Linear and exponential decreases of collision frequency with voltage are observed in the 16 and 5.7 nm pores, respectively, indicating a free energy barrier in the small pore. Moreover, the results reveal a voltage-dependent bias in the capture location toward the DNA ends, which is explained by a "pulley effect" deforming the DNA as it approaches the pore. This study provides insights into the physics governing DNA capture and translocation, which can be useful for promoting single-file translocation to enhance nanopore sensing.

Metagenomic next-generation sequencing for the identification of infections caused by Gram-negative pathogens and the prediction of antimicrobial resistance.

OBJECTIVE: The aim of this study was to evaluate the efficacy of metagenomic next-generation sequencing (mNGS) for the identification of Gram-negative bacteria (GNB) infections and the prediction of antimicrobial resistance. METHODS: A retrospective analysis was conducted on 182 patients with diagnosis of GNB infections who underwent mNGS and conventional microbiological tests (CMTs). RESULTS: The detection rate of mNGS was 96.15%, higher than CMTs (45.05%) with a significant difference (χ 2 = 114.46, P < .01). The pathogen spectrum identified by mNGS was significantly wider than CMTs. Interestingly, the detection rate of mNGS was substantially higher than that of CMTs (70.33% vs 23.08%, P < .01) in patients with but not without antibiotic exposure. There was a significant positive correlation between mapped reads and pro-inflammatory cytokines (interleukin-6 and interleukin-8). However, mNGS failed to predict antimicrobial resistance in 5 of 12 patients compared to phenotype antimicrobial susceptibility testing results. CONCLUSIONS: Metagenomic next-generation sequencing has a higher detection rate, a wider pathogen spectrum, and is less affected by prior antibiotic exposure than CMTs in identifying Gram-negative pathogens. The mapped reads may reflect a pro-inflammatory state in GNB-infected patients. Inferring actual resistance phenotypes from metagenomic data remains a great challenge.

Association between PD-L1 and Ki-67 expression and clinicopathologic features in NSCLC patients.

OBJECTIVE: To investigate the relationship between PD-L1, Ki-67 and the association between PD-L1, Ki-67, and clinicopathologic features and laboratory parameters in the peripheral blood of non-small cell lung cancer (NSCLC) patients. METHODS: The clinical records of 213 NSCLC patients were retrospectively reviewed. The patients were divided into high or low expression groups by cut-off values of Ki-67 and PD-L1. Correlation of PD-L1 and Ki-67 expression were analyzed by linear regression analysis. The data were tested by Mann-Whitney test. Relationship of PD-L1 and Ki-67 was tested by chi-square test. RESULTS: Linear regression analysis revealed a positive association between the expression of PD-L1 and Ki-67 (R2=0.26, P<0.001). The clinicopathologic features and laboratory parameters such as gender, smoking history, histological types, TNM stage, tumor size, ALB, and FIB were all significantly associated with PD-L1 and Ki-67 expression (all P<0.05). CONCLUSIONS: The expression of PD-L1 and Ki-67 is related to some clinicopathologic features and inflammatory factors, which brings new sight for exploiting combination biomarkers and therapeutic strategies.

An integrated mRNA-lncRNA signature for overall survival prediction in cholangiocarcinoma.

The combination of mRNA and lncRNA profiles for establishing an integrated mRNA-lncRNA prognostic signature has remained unexplored in cholangiocarcinoma (CCA) patients. We utilized a training dataset of 36 samples from The Cancer Genome Atlas dataset and a validation cohort (GSE107943) of 30 samples from Gene Expression Omnibus. Two mRNAs (CFHR3 and PIWIL4) and 2 lncRNAs (AC007285.1 and AC134682.1) were identified to construct the integrated signature through a univariate Cox regression (P-value = 1.35E-02) and a multivariable Cox analysis (P-value = 3.07E-02). Kaplan-Meier curve showed that patients with low risk scores had notably prolonged overall survival than those with high risk scores (P-value = 4.61E-03). Subsequently, the signature was validated in GSE107943 cohort with an area under the curve of 0.750 at 1-year and 0.729 at 3-year. The signature was not only independent from diverse clinical features (P-value = 3.07E-02), but also surpassed other clinical characteristics as prognostic biomarkers with area under the curve of 0.781 at 3-year. Moreover, the weighted gene co-expression network analysis and gene enrichment analyses found that the integrated signature were associated with metabolic-related biological process and lipid metabolism pathway, which has been implicated in the pathogenesis of CCA. Taken together, we developed an integrated mRNA-lncRNA signature that had an independent prognostic value in the risk stratification of patients with CCA.

Application of the oblique lateral interbody fusion technique in salvage surgery: technical note and case series.

Objective: The oblique lateral interbody fusion (OLIF) technique is a promising interbody fusion technique. This study summarizes the technical aspects of OLIF as a salvage surgery and the preliminary outcomes of a series of cases. Patients and methods: A retrospective review of patients with leg or back pain induced by pseudoarthrosis or adjacent segment disease after posterior lumbar interbody fusion/transforaminal lumbar interbody fusion was done. These patients underwent salvage OLIF surgeries in our institution from January 2021 to March 2022. Variables such as the demographic, clinical, surgical, and radiological characteristics of the enrolled patients were recorded and analyzed. Results: Eight patients (five females and three males; mean age 69.1 ± 5.7 years, range 63-80 years) were enrolled in this study. The mean operative time was 286.25 min (range: 230-440 min), and the estimated blood loss was 90 ml (range: 50-150 ml). Only one of the eight patients experienced a complication of lower limb motor weakness, which disappeared within 5 days after surgery. The latest data showed that the mean intervertebral space height increased from 8.36 mm preoperatively to 12.70 mm and the mean segmentary lordosis increased from 8.92° preoperatively to 15.05°. Bone fusion was achieved in all but one patient, who was followed up for only 3 months. The JOA scores Japanese Orthopaedic Association (JOA) Scores for low back pain of all patients significantly improved at the final follow-up. Conclusion: OLIF provides a safe and effective salvage strategy for patients with failed posterior intervertebral fusion surgery. Patients effectively recovered intervertebral and foraminal height with no additional posterior direct decompression.

Clinical Efficacy and Diagnostic Value of Metagenomic Next-Generation Sequencing for Pathogen Detection in Patients with Suspected Infectious Diseases: A Retrospective Study from a Large Tertiary Hospital.

Purpose: Metagenomic next-generation sequencing (mNGS) is a powerful yet unbiased method to identify pathogens in suspected infections. However, little is known about its clinical effectiveness. The present study aimed to assess the efficacy of mNGS in routine clinical practice. Patients and Methods: In this single-center retrospective cohort study, 518 patients with suspected infectious diseases were assessed for inclusion. Among them, each patient had undergone mNGS testing; 407 patients had undergone both microbial culture and mNGS testing. The result of mNGS testing was compared to microbial culture performed concurrently. The diagnostic performance of mNGS was evaluated using the comprehensive clinical diagnosis as the reference standard. Results: There was a significant difference in the positive detection rates of pathogens between mNGS and culture (331/407, 81.3% vs 79/407, 19.4%, P < 0.001). The sensitivity of mNGS was much higher than the culture method (79.5% vs 21.3%, P < 0.001), especially in sample types of sputum and bronchoalveolar lavage fluid (BALF). Notably, the sensitivity of blood mNGS was relatively lower than other sample types (67.4% vs 88.9-93.8%). Pathogen cfDNA load based on standardized stringently mapped read number at the species level of microorganisms (SDSMRN) was significantly lower in blood than in other sample types from the same patient (P = 0.0003). Importantly, mNGS directly led to a change of treatment regimen in 142 (27.4%) cases, including antibiotic escalation (15.3%), antibiotic de-escalation (9.1%), and early definitive diagnosis to initiate appropriate treatment (3.1%). Conclusion: Our in-house mNGS platform significantly improved the sensitivity for the diagnosis of infectious diseases. mNGS has the potential to improve clinical outcomes by optimizing antimicrobial therapy.

Comparison of Lumbar Fusion Surgical Outcomes Between Patients with Lumbar Spinal Stenosis ≥80 Versus 65-79 Years Old.

BACKGROUND The efficacy of lumbar fusion surgery in patients age 80 years and older with lumbar spinal stenosis (LSS) is still controversial. This retrospective study aimed to evaluate the surgical outcomes of LSS patients ³80 vs 65-79 years. MATERIAL AND METHODS The study included 66 patients diagnosed with LSS from 2014 to 2020; 33 patients were ³80 years and 33 patients were 65-79 years. The 2 groups were matched for sex and surgical segment. All patients underwent posterior lumbar decompression, fixation, and fusion surgery. The Numerical Rating Scale (NRS) of leg and back pain, Oswestry Disability Index (ODI), Short-Form Health Survey (SF-36), and radiographic data were collected before surgery and at 3, 6, and 12 months postoperatively. RESULTS NRS (back) in the ≥80 years group was significantly higher than in the 65-79 years group at 3 months [2 (0-3) vs 1 (0-3), P=0.001]. Improvement of SF-36 (3 months: 15.7±4.9 vs 27.6±5.4, P<0.001; 6 months: 27.3±6.8 vs 31.5±5.6, P=0.011) and Physical Component Score (PCS) (3 months: 6.5±2.5 vs 17.0±3.6, P<0.001; 6 months: 15.9±3.4 vs 20.1±3.1, P<0.001) at 3 and 6 months in the ≥80 years group were significantly smaller than in 65-79 years group. There was a difference of Pfirrmann index of adjacent segment disc between the 2 groups at 12 months [≥80 vs 65-79 years group: 5.5 (4-8) vs 5 (3-8), P=0.003]. CONCLUSIONS Lumbar fusion surgery in patients ≥80 years with LSS can provide comparable improvements in clinical and radiographic outcomes compared with younger patients. Postoperative physiological function recovery was slower in patients ≥80 years.

A rare discal cyst with concurrent double-level isthmic lumbar spondylolisthesis: a case report and literature review.

We presented a 47-year old male diagnosed as a discal cyst with concurrent isthmic lumbar spondylolisthesis, which was not reported in the literature previously. A cystectomy with two-segmental transforaminal lumbar interbody fusion and instruments was performed. The association between concurrent discal cyst and isthmic spondylolisthesis may be illuminated by excessive strain and focal degeneration of the disc. Preoperative discography is probably unnecessary when surgical resection and histopathology are indicated for the patient.

Surgical Precision and Efficiency of a Novel Electromagnetic System Compared to a Robot-Assisted System in Percutaneous Pedicle Screw Placement of Endo-LIF.

STUDY DESIGN: Retrospective database study. OBJECTIVES: To compare the accuracy and safety of 2 types of a computer-assisted navigation system for percutaneous pedicle screw placement during endoscopic lumbar interbody fusion. METHODS: From May 2019 to January 2020, data of 56 patients who underwent Endo-LIF with a robot-assisted system and with an electromagnetic navigation system were compared. The pedicles in all patients were subjected to postoperative CT scan to assess screw correction by measuring the perpendicular distance between the pedicle cortical wall and the screw surface. The registration and matching time, guide-wire insertion time, the entire surgery time, and X-ray exposure time were recorded. RESULTS: In the robot-assisted group, 25 cases with 100 percutaneous pedicle screws were included, and the excellent and good rate was 95%. In the electromagnetic navigation group, 31 cases with 124 screws were included, and the excellent rate was 97.6%. There was no statistical difference between the two groups (P > 0.05). The registration time and the total time for the surgery also showed no statistical differences (P > 0.05). The main difference between the two groups was the guide-wire insertion time and the X-ray exposure time (P < 0.05). CONCLUSIONS: Both electromagnetic navigation and robot-assisted are safe and efficient for percutaneous pedicle screw placement. Electromagnetic navigation system has obvious advantages over robot-assisted in terms of faster guide-wire placement and less X-ray exposure. Robot-assisted for percutaneous pedicle screw placement offers a preoperative planning system and a stable registration system, with obvious drawbacks of a strict training curve.

Fully Endoscopic 360° Decompression for Central Lumbar Spinal Stenosis Combined with Disc Herniation: Technical Note and Preliminary Outcomes of 39 Cases.

Purpose: To evaluate the outcomes, feasibility, and safety of endoscopic unilateral laminectomy, bilateral decompression and discectomy (Endo-ULBDD) for central lumbar spinal stenosis (CLSS) combined with disc herniation (DH). Methods: This study includes 39 patients diagnosed with CLSS combined with DH who met the inclusion criteria and underwent surgery for Endo-ULBDD from April 2020 to March 2021. The mean age of the patients, operation time, hospitalization time, time in bed, and complications were recorded. Patients were followed up for at least 12 months. Visual analog scale (VAS) scores for low-back and lower-limb pain and Oswestry Disability Index (ODI) scores were evaluated preoperatively, before discharge, and at 3, 6, and 12 months postoperatively. To evaluate clinical effectiveness 12 months postoperatively, the modified MacNab criteria were used. Results: The mean age of the patients was 59.9 years, the mean operation time was 82.1 minutes, the mean hospitalization time was 3.7 days, and the mean time in bed was 20.9 hours. The mean VAS scores of low-back and lower-limb pain improved from 5.9 and 7.2 to 2.0 and 1.6, respectively (P < 0.05). The ODI score improved from 56.0 to 16.7 (P < 0.05). The overall excellent-good rate of the modified MacNab criteria was 89.7%. Two kinds of complications occurred in 4 patients (10.3%), including 1 patient whose inferior articular process was excessively removed and 3 patients who suffered from postoperative dysesthesia. No other severe complications were noted. Conclusion: Endo-ULBDD is a safe, feasible, efficient, and minimally invasive approach to treating CLSS combined with DH.

Expression of Potato StDRO1 in Arabidopsis Alters Root Architecture and Drought Tolerance.

Potato (Solanum tuberosum L) is the third important crop for providing calories to a large human population, and is considered sensitive to moderately sensitive to drought stress conditions. The development of drought-tolerant, elite varieties of potato is a challenging task, which can be achieved through molecular breeding. Recently, the DEEPER ROOTING 1 (DRO1) gene has been identified in rice, which influences plant root system and regulates grain yield under drought stress conditions. The potato StDRO1 protein is mainly localized in the plasma membrane of tobacco leaf cells, and overexpression analysis of StDRO1 in Arabidopsis resulted in an increased lateral root number, but decreased lateral root angle, lateral branch angle, and silique angle. Additionally, the drought treatment analysis indicated that StDRO1 regulated drought tolerance and rescued the defective root architecture and drought-tolerant phenotypes of Atdro1, an Arabidopsis AtDRO1 null mutant. Furthermore, StDRO1 expression was significantly higher in the drought-tolerant potato cultivar "Unica" compared to the drought-sensitive cultivar "Atlantic." The transcriptional response of StDRO1 under drought stress occurred significantly earlier in Unica than in Atlantic. Collectively, the outcome of the present investigation elucidated the role of DRO1 function in the alternation of root architecture, which potentially acts as a key gene in the development of a drought stress-tolerant cultivar. Furthermore, these findings will provide the theoretical basis for molecular breeding of drought-tolerant potato cultivars for the farming community.

Comparison of electromagnetic and optical navigation assisted Endo-TLIF in the treatment of lumbar spondylolisthesis.

Uniportal full endoscopic posterolateral transforaminal lumbar interbody fusion (Endo-TLIF) with percutaneous pedicle screw fixation is a promising, minimally invasive method for the treatment of lumbar spondylolisthesis. However, repeated radiation exposure from X-rays and the steep learning curve remain to be improved.

Application of electromagnetic navigation in endoscopic transforaminal lumbar interbody fusion: a cohort study.

STUDY DESIGN: Clinical retrospective cohort study. OBJECTIVES: To explore the application of the electromagnetic navigation system in Endo-TLIF. MATERIALS AND METHODS: From May 2019 to March 2020, 76 patients with single-segment lumbar spondylolisthesis treated by electromagnetic navigation-assisted Endo-TLIF (NE group) and conventional Endo-TLIF (CE group) were enrolled in the study. Time of pedicle screw implantation, entire operation time, the number of intraoperative X-ray fluoroscopy exposures, total blood loss, incision length, ambulation time, accuracy of pedicle screws, complications, visual analog scale for back and leg pain, Oswestry Disability Index, Japanese Orthopedic Association score and postoperative fusion rates were recorded, respectively. RESULTS: There were no significant differences in preoperative demographics between the NE and CE groups (P > 0.05). The mean number of intraoperative X-ray fluoroscopy exposures, guidewires insertion, entire operation time, total blood loss and adjustment rate of screws in the NE group were significantly less compared with the CE group (P < 0.05, respectively). There were no significant differences in clinical parameters between the two groups at different time points in the follow-up period (P > 0.05). There was no statistical difference in fusion rates between the two groups. In addition, one case of cage subsidence was observed after surgery in the CE group. CONCLUSION: Electromagnetic navigation systems could be applied throughout the entire surgical course and ameliorate the shortcomings of the conventional Endo-TLIF technique to reduce radiation exposure, improve accuracy, avoid repetitive operations and shorten surgery time and the required learning curve of the procedure. LEVEL OF EVIDENCE I: Diagnostic: individual cross-sectional studies with consistently applied reference standard and blinding.

Identification and Expression Analysis of Calcium-Dependent Protein Kinases Gene Family in Potato Under Drought Stress.

Calcium-dependent protein kinases (CDPKs) are a class of serine/threonine protein kinases encoded by several gene families that play key roles in stress response and plant growth and development. In this study, the BLAST method was used to search for protein sequences of the potato Calcium-dependent protein kinase gene family. The chromosome location, phylogeny, gene structures, gene duplication, cis-acting elements, protein-protein interaction, and expression profiles were analyzed. Twenty-five CDPK genes in the potato genome were identified based on RNA-seq data and were clustered into four groups (I-IV) based on their structural features and phylogenetic analysis. The result showed the composition of the promoter region of the StCDPKs gene, including light-responsive elements such as Box4, hormone-responsive elements such as ABRE, and stress-responsive elements such as MBS. Four pairs of segmental duplications were found in StCDPKs genes and the Ka/Ks ratios were below 1, indicating a purifying selection of the genes. The protein-protein interaction network revealed defense-related proteins such as; respiratory burst oxidase homologs (RBOHs) interacting with potato CDPKs. Transcript abundance was measured via RT-PCR between the two cultivars and their relative expression of CDPK genes was analyzed after 15, 20, and 25 days of drought. There were varied expression patterns of StCDPK3/13/21 and 23, between the two potato cultivars under mannitol induced-drought conditions. Correlation analysis showed that StCDPK21/22 and StCDPK3 may be the major differentially expressed genes involved in the regulation of malondialdehyde (MDA) and proline content in response to drought stress, opening a new research direction for genetic improvement of drought resistance in potato.

Relationship between hinge fracture and postoperative axial symptoms after cervical laminoplasty.

OBJECTIVE: To identify the clinical characteristics of fractured hinges after open-door cervical laminoplasty for cervical canal stenosis and explore the relationship between hinge fractures and axial symptoms. METHODS: This was a retrospective study of patients with cervical myelopathy who underwent open-door laminoplasty between November 2014 and November 2016 at the Affiliated Hospital of Qingdao University. Cervical CT scans were performed after surgery and the Takeuchi criteria were applied to evaluate the postoperative axial symptoms. RESULTS: Of 223 opened laminae in 67 patients, 67 laminae (30.0%) in 30 patients (44.8%) showed fracture. The frequency of hinge fractures was higher at C6 (53.7%). Forty-nine fractured laminae (73.13%) were non-displaced and 18 were displaced. At 3 months, 33 fractured laminae (49.3%) showed bony union on CT, and union rates were 86.6% and 91.0% at 6 and 12 months, respectively, indicating that the union rate was lower for displaced fractures than for non-displaced fractures. Among the 67 patients, 14 had axial symptoms: three of 37 (8.1%) patients without hinge fractures and 11 of 30 (36.7%) patients with hinge fractures. One year later, the hinge fractures were healed in 24/30 patients. Among the six unhealed patients, five still suffered from axial symptoms. The frequency of axial symptoms was higher in the patients with three or more hinge fractures (66.7%) than in the patients with only one (16.7%) or two (46.7%) hinge fractures. CONCLUSIONS: Patients with hinge fractures may have an increased risk for axial symptoms after open-door cervical laminoplasty. The frequency of axial symptoms decreases with fracture healing.

Hsa_circ_0134111 promotes intervertebral disc degeneration via sponging miR-578.

Intervertebral disc degeneration (IDD) is a chronic degenerative and age-dependent process characterized by aberrant apoptosis, proliferation, synthesis, and catabolism of the extracellular matrix of the nucleus pulposus (NP) cells. Recently, studies showed that circular RNAs play important roles in the development of many diseases. However, the role of circRNAs in IDD development remains unknown. We showed that circ_0134111 level was overexpressed in IDD tissue samples as compar-ed to control tissues. The upregulation of circ_0134111 was more drastic in the moderate and severe IDD cases than in those with mild IDD. In addition, we showed that interleukin-1ß and tumor necrosis factor-α exposure significantly enhanced circ_0134111 expression in NP cells. Furthermore, ectopic expression of circ_0134111 induced proliferation, pro-inflammatory cytokine secretion, and ECM degradation in the NP cells. We also showed that circ_0134111 directly interacted with microRNA (miR)-578 in NP cells where elevated expression of circ_0134111 enhanced the ADAMTS-5 and MMP-9 expression. Moreover, miR-578 expression was significantly decreased in IDD patients and the miR-578 expression was negatively correlated with circ_0134111 expression in the IDD samples. Interleukin-1ß and tumor necrosis factor-α exposure significantly decreased miR-578 levels in NP cells, in which ectopic miR-578 expression inhibited cell growth, pro-inflammatory cytokine expression, and ECM degradation. Finally, we showed that circ_0134111 overexpression induced the IDD-related phenotypic changes through inhibiting miR-578. These data suggested that circ_0134111 could promote the progression of IDD through enhancing aberrant NP cell growth, inflammation, and ECM degradation partly via regulating miR-578.

Sequential endoscopic and robot-assisted surgical solutions for a rare fungal spondylodiscitis, secondary lumbar spinal stenosis, and subsequent discal pseudocyst causing acute cauda equina syndrome: a case report.

BACKGROUND: Fungal spondylodiscitis is a rare infectious disease. The secondary lumbar spinal stenosis and postoperative discal pseudocyst were even rarer. The surgical interventions were disputed, yet endoscopic and robot-assisted techniques may be helpful under different circumstances. CASE PRESENTATION: A 62-year-old female was diagnosed as infectious spondylodiscitis at the L4/5 level and a posterolateral endoscopic debridement was performed after invalid conservative therapy. Causative organism culture revealed a rare fungus, Candida tropicalis. A secondary spinal stenosis with refractory radiculopathy occurred almost 3 years after the first surgery and a successful endoscopic surgery was implemented aiming to decompress the nerve in a minimally invasive way. However, 2 months later, the patient manifested severe acute cauda equina syndrome and radiological examinations suggested a rare postoperative discal pseudocyst. A laminectomy followed by a pseudocystectomy was applied to achieve thorough decompression. An innovative double trajectory system (simultaneous traditional pedicle screw and cortical bone trajectory screw) accompanied by posterolateral fusion was designed and executed by the professional robot-assisted system. CONCLUSION: Endoscopic and robot-assisted techniques may provide alternative solutions for fungal spondylodiscitis and accompanied sequelae.

Inhibition of RhoA/MRTF-A signaling alleviates nucleus pulposus fibrosis induced by mechanical stress overload.

PURPOSE/AIM: : Intervertebral disc degeneration (IDD) is the leading cause of lower back pain, and clinically useful drugs for IDD are unavailable. Mechanical stress overload-induced fibrosis plays a critical role in IDD. RhoA/MRTF-A signaling is known to regulate tissue fibrosis; however, the effect of RhoA/MRTF-A on the development of IDD is unclear. MATERIALS AND METHODS: : The expression of aggrecan, collagen I, collagen II, MMP-12, CTGF, and MRTF-A in nucleus pulposus (NP) samples from IDD patients and controls was detected by immunohistochemical staining. Primary nucleus pulposus cells (NPCs) were isolated and cultured to establish an overload strain model treated with or without CCG-1423. The protein levels of RhoA, ROCK2, MRTF-A, CTGF, and MMP-12 as well as fibrosis-associated proteins were detected by western blotting and immunofluorescence. RESULTS: : Collagen I, MMP-12, and CTGF were significantly upregulated, and aggrecan and collagen II were significantly downregulated in the IDD samples. The cellular localization of MRTF-A was associated with intervertebral disc (IVD) degeneration. Overloaded strain enhanced the nuclear translocation of MRTF-A and changed the NPC morphology from spindle-shaped to long strips. Additional experiments showed that RhoA, ROCK2, MRTF-A, SRF, MMP-12, and CTGF were upregulated; however, aggrecan and collagen II were downregulated in NPCs under overload strain. CCG-1423, a RhoA/MRTF-A pathway inhibitor, reversed strain-induced fibrosis. CONCLUSION: : Mechanical stress activates RhoA/MRTF-A signaling to promote extracellular matrix (ECM) degeneration in the NP, which is associated with the development of IDD. Our findings suggest that the RhoA/MRTF-A inhibitor CCG-1423 can alleviate NPC degeneration caused by overload stress and has potential as a therapeutic agent for IDD.