Ruxolitinib improves the inflammatory microenvironment,restores glutamate homeostasis,and promotes functional recovery after spinal cord injury

The inflammatory microenvironment and neurotoxicity can hinder neuronal regeneration and functional recovery after spinal cord injury.Ruxolitinib,a JAK-STAT inhibitor,exhibits effectiveness in autoimmune diseases,arthritis,and managing inflammatory cytokine storms.Although studies have shown the neuroprotective potential of ruxolitinib in neurological trauma,the exact mechanism by which it enhances functional recovery after spinal cord injury,particularly its effect on astrocytes,remains unclear.To address this gap,we established a mouse model of T10 spinal cord contusion and found that ruxolitinib effectively improved hindlimb motor function and reduced the area of spinal cord injury.Transcriptome sequencing analysis showed that ruxolitinib alleviated inflammation and immune response after spinal cord injury,restored EAAT2 expression,reduced glutamate levels,and alleviated excitatory toxicity.Furthermore,ruxolitinib inhibited the phosphorylation of JAK2 and STAT3 in the injured spinal cord and decreased the phosphorylation level of nuclear factor kappa-B and the expression of inflammatory factors interleukin-1β,interleukin-6,and tumor necrosis factor-α.Additionally,in glutamate-induced excitotoxicity astrocytes,ruxolitinib restored EAAT2 expression and increased glutamate uptake by inhibiting the activation of STAT3,thereby reducing glutamate-induced neurotoxicity,calcium influx,oxidative stress,and cell apoptosis,and increasing the complexity of dendritic branching.Collectively,these results indicate that ruxolitinib restores glutamate homeostasis by rescuing the expression of EAAT2 in astrocytes,reduces neurotoxicity,and effectively alleviates inflammatory and immune responses after spinal cord injury,thereby promoting functional recovery after spinal cord injury.

Extradural contralateral S1 nerve root transfer for spastic lower limb paralysis

The current study aims to ascertain the anatomical feasibility of transferring the contralateral S1 ventral root(VR)to the ipsilateral L5 VR for treating unilateral spastic lower limb paralysis.Six formalin-fixed(three males and three females)cadavers were used.The VR of the contralateral S1 was transferred to the VR of the ipsilateral L5.The sural nerve was selected as a bridge between the donor and recipient nerve.The number of axons,the cross-sectional areas and the pertinent distances between the donor and recipient nerves were measured.The extradural S1 VR and L5 VR could be separated based on anatomical markers of the dorsal root ganglion.The gross distance between the S1 nerve root and L5 nerve root was 31.31(±3.23)mm in the six cadavers,while that on the diffusion tensor imaging was 47.51(±3.23)mm in 60 patients without spinal diseases,and both distances were seperately greater than that between the outlet of S1 from the spinal cord and the ganglion.The numbers of axons in the S1 VRs and L5 VRs were 13414.20(±2890.30)and 10613.20(±2135.58),respectively.The cross-sectional areas of the S1 VR and L5 VR were 1.68(±0.26)mm2 and 1.08(±0.26)mm2,respectively.In conclusion,transfer of the contralateral S1 VR to the ipsilateral L5 VR may be an anatomically feasible treatment option for unilateral spastic lower limb paralysis.

Involvement of 2'-5'oligoadenylate synthetase-like proteinin the survivalof Mycobacterium tuberculosis avirulent strain in macrophages

Mycobacterium tuberculosis(M.tuberculosis)can replicate in the macrophage by interfering with many host protein functions.While it is far from known these host proteins for controlling M.tuberculosis infection.Herein,we infected macrophages including THP-1 and Raw264.7 cells with M.tuberculosis and identified the differentially expressed genes(DEGs)in the interferon signaling pathway.Among them,2'-5'oligoadenylate synthetase-like(OASL)underwent the greatest upregulation in M.tuberculosis-infected macrophages.Knockdown of the expression of OASL attenuated M.tuberculosis survival in macrophages.Further,bioinformatics analysis revealed the potential interaction axis of OASL-TAB3-RvO127,which was further validated by the yeast-two-hybrid(Y2H)assay and Co-IP.This interaction axis might regulate the M.tuberculosis survival and proliferation in macrophages.The study reveals a possible role of OASL during M.tuberculosis infection as a target to control its propagation.

Surgical outcomes of mini-open Wiltse approach and conventional open approach in patients with single-segment thoracolumbar fractures without neurologic injury

This study aimed to introduce a novel mini-open pedicle screw fixation technique via Wiltse approach, and com- pared it with the traditional posterior open method. A total of 72 cases of single-segment thoracolumbar fractures without neurologic injury underwent pedicle screw fixation via two different approaches. Among them, 37 patients were treated using posterior open surgery, and 35 patients received mini-open operation via Wiltse approach. Crew placement accuracy rate, operative time, blood loss, postoperative drainage, postoperative hospitalization time, radiation exposure time, postoperative improvement in R value, Cobb＇s angle and visual analog scale （VAS） scores of the two methods were compared. There were no significant differences in the accuracy rate of pedicle screw placement, radiation exposure and postoperative R value and Cobb＇s angle improvement between the two groups. However, the mini-open method had obvious advantages over the conventional open method in operative time, blood loss, postoperative drainage, postoperative hospitalization time, and postoperative improvement in VAS. The mini-open pedicle screw technique could be applied in treatment of single-segment thoracolumbar fracture without neurologic injury and had advantages of less tissue trauma, short operative and rehabilitative time on the premise of guaranteed accuracy rate and no increased radiation exposure.

Position and complications of pedicle screw insertion with or without image-navigation techniques in the thoracolumbar spine:a meta-analysis of comparative studies

Computer-navigated pedicle screw insertion is applied to the thoracic and lumbar spine to attain high insertion accuracy and a low rate of screw-related complications.However,some in vivo and in vitro studies have shown that no advantages are gained with the use of navigation techniques compared to conventional techniques.Additionally,inconsistent conclusions have been drawn in various studies due to different population characteristics and methods used to assess the accuracy of screw placement.Moreover,it is not clear whether pedicle screw insertion with navigation techniques decreases the incidence of screw-related complications.Therefore,this study was sought to perform a meta-analysis of all available prospective evidence regarding pedicle screw insertion with or without navigation techniques in human thoracic and lumbar spine.We considered in vivo comparative studies that assessed the results of pedicle screw placement with or without navigation techniques.PubMed,Ovid MEDLINE and EMBASE databases were searched.Three published randomized controlled trials（RCTs） and nine retrospective comparative studies met the inclusion criteria.These studies included a total of 732 patients in whom 4,953 screws were inserted.In conclusion,accuracy of the position of grade I,II,III and IV screws and complication rate related to pedicle screw placement were significantly increased when navigation techniques were used in comparison to conventional techniques.Future research in this area should include RCTs with well-planned methodology to limit bias and report on validated,patient-based outcome measures.

A new free-hand pedicle screw placement technique with reference to the supraspinal ligament

We sought to compare the safety and accuracy of a new free-hand pedicle screw placement technique to that of the conventional technique. One hundred fifty-three consecutive adult patients with simple fracture in the tho- racic or/and lumbar spine were alternately assigned to either the new free-hand or the conventional group. In the new free-hand technique group, preoperative computerized tomography （CT） images were used to calculate the targeted medial-lateral angle of each pedicle trajectory and the pedicle screw was inserted perpendicular to the corresponding supraspinal ligament. In the conventional technique group, the medial-lateral and cranial-caudal angle of each pedicle trajectory was determined by intraoperatively under fluoroscopic guidance. The accuracy rate of pedicle screw placement, the time of intraoperative fluoroscopy, the operating time and the amount of blood loss during operation were respectively compared. All screws were analyzed by using intraoperative ra- diographs, intraoperative triggered electromyography （EMG） monitoring data, postoperative CT data and clinical outcomes. The accuracy rate of pedicle screw placement in the new free-hand technique group and the conven- tional technique group was 96.3% and 94.2% （P 〈 0.05）, respectively. The intraoperative fluoroscopy time of the new technique group was less than that of the conventional technique group （5.37 seconds vs. 8.79 seconds, P 〈 0.05）. However, there was no statistical difference in the operating time and the amount of blood loss during op- eration （P 〉 0.05）. Pedicle screw placement with the free-hand technique which keeps the screw perpendicular to the supraspinal ligament is an accurate, reliable and safe technique to treat simple fracture in the thoracic or lum- bar spine.

Comparison of the modified Wiltse’s approach with spinal minimally invasive system and traditional approach for the therapy of thoracolumbar fracture

Thoracolumbar fractures are usually treated by open posterior pedicle screw fixation.However,this procedure involves massive paraspinal muscle stripping,inflicting surgical trauma,and prolonged X-ray exposure.In this study,we observed 127 patients with single-segment injury thoracolumbar fractures.Thirty-six patients were treated by the modified Wiltse’s paraspinal approach with minimally invasive channel system,while 91 patients were treated via traditional posterior approach.Operation time,intraoperative blood loss,intraoperative fluoroscopy frequency,screw placement accuracy,visual analogue scale score,and Cobb’s angle of two groups were compared.The X-ray exposure times were notably reduced(4.2±1.6) in the new approach group(P<0.05).The pedicle screw placement accuracy and Cobb’s angle after surgery were similar in the two groups.We conclude that modified Wiltse’s paraspinal approach w ith spinal minimally invasive channel system surgery can significantly reduce the X-ray exposure times and is an alternative therapy for the thoracolumbar fracture.

Neuroanatomy and clinical analysis of the cervical sympathetic trunk and longus colli

Anterior cervical surgery is commonly used for cervical vertebral body lesions. However, the structure of blood vessels and nerve tissues along the route of anterior cervical surgery is complex. We aimed to measure the data of the longus colli, the sympathetic trunk and the cervical sympathetic trunk （CST） ganglia in Chinese cadaver speci- mens. A total of 32 adult cadavers were studied. We delineated the surgical anatomy of the CST. The superior and inferior/cervicothoracic ganglia of the sympathetic trunk consistently appeared. The middle ganglion was observed in 28.1% of the specimens and there were 2 cases of unilateral double middle cervical ganglia. The inferior ganglion was observed in 25.0% of the specimens and the cervicothoracic ganglion was observed in the remaining speci- mens. The distance between the CST gradually decreased from the top to the bottom, and the distance between the medial edges of the longus colli gradually broadened from the top down. The average angle between the bilat- eral CST and the midline of the vertebra was 11.2°± 1.8° on the left side and 10.3°± 1.4° on the right side. The average angle between the medial margins of longus colli of both sides was 11.1°± 1.9°. The CST is at high risk when LC muscle is cut transversely or is dragged heavily, especially at the levels of C6 and C7. Awareness of the regional anatomy of the CST could help surgeons to identify and preserve it during anterior cervical surgeries.

A comparison of robot-assisted and fluoroscopy-assisted kyphoplasty in the treatment of multi-segmental osteoporotic vertebral compression fractures

Osteoporotic vertebral compression fracture(OVCF)has become a major public health issue that becomes more pressing with increasing global aging.Percutaneous kyphoplasty(PKP)is an effective treatment for OVCF.Robot-assisted PKP has been utilized in recent years to improve accuracy and reduce complications.However,the effectiveness of robot-assisted PKP in the treatment of multi-segmental OVCF has yet to be proved.This study was designed to compare the efficacy of robot-assisted and conventional fluoroscopy-assisted multi-segmental PKP.A total of 30 cases with multi-segmental OVCF between April 2019 and April 2021 were included in this study.Fifteen cases were assigned to the robot-assisted PKP group(robot group)and 15 cases to the conventional fluoroscopy-assisted PKP group(conventional fluoroscopy group).The number of fluoroscopic exposures,fluoroscopic dose,operation time,cement leakage rate,visual analog scale(VAS)score,vertebral kyphosis angle(VKA),and height of fractured vertebral body(HFV)were compared between the 2 groups.The number of fluoroscopic exposures,fluoroscopic doses,and cement leakage rates in the robot group were lower than in the conventional fluoroscopy group(P<0.05)while the operative time in the robot group was longer than in the conventional fluoroscopy group(P<0.05).VAS score and VKA were decreased and HFV was increased after surgery in both groups(P<0.05).Therefore,robot-assisted PKP for the treatment of multi-segmental OVCF can reduce the number of fluoroscopic exposures,fluoroscopic doses,and cement leakage compared to conventional treatment.As such,robot-assisted PKP has good application prospects and is potentially more effective in the treatment of multi-segmental OVCF.

Tacrolimus reduces scar formation and promotes sciatic nerve regeneration

A sciatic nerve transection and repair model was established in Sprague-Dawley rats by transecting the tendon of obturator internus muscle in the greater sciatic foramen and suturing with nylon sutures. The models were treated with tacrolimus gavage （4 mg/kg per day） for 0, 2, 4 and 6 weeks. Specimens were harvested at 6 weeks of intragastric administration. Masson staining revealed that the collagen fiber content and scar area in the nerve anastomosis of the sciatic nerve injury rats were significantly reduced after tacrolimus administration. Hematoxylin-eosin staining showed that tacrolimus significantly increased myelinated nerve fiber density, average axon diameter and myelin sheath thickness. Intragastric administration of tacrolimus also led to a significant increase in the recovery rate of gastrocnemius muscle wet weight and the sciatic functional index after sciatic nerve injury. The above indices were most significantly improved at 6 weeks after of tacrolimus gavage. The myelinated nerve fiber density in the nerve anastomosis and the sciatic nerve functions had a significant negative correlation with the scar area, as detected by Spearman＇s rank correlation analysis. These findings indicate that tacrolimus can promote peripheral nerve regeneration and accelerate the recovery of neurological function through the reduction of scar formation.

Rapid identification of spinal ventral and dorsal roots using a quartz crystal microbalance

The fast and accurate identification of nerve tracts is critical for successful nerve anastomosis. Taking advantage of differences in acetylcholinesterase content between the spinal ventral and dorsal roots, we developed a novel quartz crystal microbalance method to distinguish between these nerves based on acetylcholinesterase antibody reactivity. The acetylcholinesterase antibody was immobilized on the electrode surface of a quartz crystal microbalance and reacted with the acetylcholinesterase in sample solution. The formed antigen and antibody complexes added to the mass of the electrode inducing a change in frequency of the electrode. The spinal ventral and dorsal roots were distinguished by the change in frequency. The ventral and dorsal roots were cut into 1 to 2-mm long segments and then soaked in 250 pL PBS. Acetylcholinesterase antibody was immobilized on the quartz crystal microbalance gold electrode surface. The results revealed that in 10 minutes, both spinal ventral and dorsal roots induced a frequency change; however, the frequency change induced by the ventral roots was notably higher than that induced by the dorsal roots. No change was induced by bovine serum albumin or PBS. These results clearly demonstrate that a quartz crystal microbalance sensor can be used as a rapid, highly sensitive and accurate detection tool for the quick identification of spinal nerve roots intraoperatively.

Potential risk of mitomycin C at high concentrations on peripheral nerve structure

Although the local application of mitomycin C may prevent epidural adhesion after laminectomy, mitomycin C can induce neurotoxicity in optic and acoustic nerves at high concentrations. To determine the safe concentration range for mitomycin C, cotton pads soaked with mitomycin C at different concentrations （0.1, 0.3, 0.5, and 0.7 mg/mL） were immediately applied for 5 minutes to the operation area of rats that had undergone laminectomy at L1. Rat sciatic nerves, instead of dorsal nerves, were used in this study. The results showed that mitomycin C at 0.1-0.5 mg/mL did not damage the structure and function of the sciatic nerve, while at 0.7 mg/mL, mitomycin C signiifcantly reduced the thickness of the sciatic nerve myelin sheath compared with lower concen-trations, though no functional change was found. These experimental ifndings indicate that the local application of mitomycin C at low concentrations is safe to prevent scar adhesion following laminectomy, but that mitomycin C at high concentrations （＆gt;0.7 mg/mL） has potential safety risks to peripheral nerve structures.

A novel entry point for pedicle screw placement in the thoracic spine

This study was aimed to introduce a novel entry point for pedicle screw fixation in the thoracic spine and compare it with the traditional entry point. A novel entry point was found with the aim of improving accuracy, safety and stability of pedicle screw technique based on anatomical structures of the spine. A total of 76 pieces of normal thoracic CT images at the transverse plane and the thoracic pedicle anatomy of 6 cadaveric specimens were recruited.Transverse pedicle angle（TPA）, screw length, screw placement accuracy rate and axial pullout strength of the two different entry point groups were compared. There were significant differences in the TPA, screw length, and the screw placement accuracy rate between the two groups（P〈0.05）. The maximum axial pullout strength of the novel entry point group was slightly larger than that of the traditional group. However, the difference was not significant（P 〉 0.05）. The novel entry point significantly improved the accuracy, stability and safety of pedicle screw placement.With reference to the advantages above, the new entry point can be used for spinal internal fixations in the thoracic spine.

Effects of leptin on femoral fracture in rats

In this study, our objective was to evaluate effects of leptin on fracture healing in rats. Seventy two male SpragueDawley（SD） rats were randomized into 3 groups. Standardized femoral fractures were created in all the rats. Group A was treated with 1 mL normal saline（NS）, group B with 0.3μg/kg leptin in 1 mL NS, and group C with 0.5μg/kg leptin in 1 mL NS for 2 weeks intraperitoneally. Each group was divided into three subgroups including 8 rats for evaluation at 2, 4 and 8 weeks. Radiological evaluation showed that callus formation of group B and C was all significantly higher than group A at 8 weeks（P=0.04 and P = 0.013, respectively）. There was no statistically significant difference in fracture healing between group B and group C at 8 weeks（P = 0.197）. Histological evaluation revealed fracture healing of group B and C was better than group A at 4 weeks（P=0.01 and P= 0.002,respectively） and 8 weeks（P = 0.008 and P= 0.003, respectively）. Micro-computed tomography（Micro-CT） analysis demonstrated that greater amounts of bony callus and evidence of bone fusion were observed in group B and C at 4 weeks（P = 0.02 and P = 0.04, respectively） and 8 weeks（P= 0.005 and P = 0.001, respectively） compared to group A. Group C also had better fracture healing than group B at 8 weeks（P = 0.01）. In conclusion, leptin has a positive effect on rat femoral fracture healing.

A novel extradural nerve transfer technique by coaptation of C4 to C5 and C7 to C6 for treating isolated upper trunk avulsion of the brachial plexus

The study aimed to demonstrate the feasibility of an extradural nerve anastomosis technique for the restoration of a C5 and C6 avulsion of the brachial plexus.Nine fresh frozen human cadavers were used.The diameters,sizes,and locations of the extradural spinal nerve roots were observed.The lengths of the extradural spinal nerve roots and the distance between the neighboring nerve root outlets were measured and compared in the cervical segments.In the spinal canal,the ventral and dorsal roots were separated by the dura and arachnoid.The ventral and dorsal roots of C7 had sufficient lengths to anastomose those of C6.The ventral and dorsal of C4 had enough length to be transferred to those of C5,respectively.The feasibility of this extradural nerve anastomosis technique for restoring C5 and C6 avulsion of the brachial plexus in human cadavers was demonstrated in our anatomical study.

DIC imaging for identification of motor and sensory nerves

Identification of motor and sensory nerves is important in applications such as nerve injury repair.Conventional practice relies on time consuming staining methods for this purpose.Here,we use laser scanning infrared diferential interference contrast(IR-DIC)microscopy for label-free observation of the two types of nerve.Ventral and dorsal nerve roots of adult beagle dogs were collected and sections of different thicknesses were imaged with an IR-DIC microscope.Different texture patterns of the IR-DIC images of the motor and sensory nerve can be distinguished when the section thickness increases to 40 pm.This suggests that nerve fibers in motor and sensory nerves have different distribution patterns.The result hints a potential new way for more rapid identification of nerve type in peripheral nerve repair surgery.

Perturbation Solutions for Annular Flow of Small Gap

The perturbation method is used to solve the control equations of a three-dimensional annular flow inside a small gap. The nonlinear equations are separated into zeroth-order and first-order perturbation equations. The velocity and pressure distributions are solved successively by different numerical methods with the zeroth-order and first-order equation. Agreement in results is found with the present method and software ANSYS-CFX, which illustrates the applicability of perturbation method in solving complicated flow field inside small gaps.

Single-cell transcriptome landscape and antigen receptor dynamic during SARS-CoV-2 vaccination

Vaccination by inactivated vaccine is an effective strategy to prevent the COVID-19pandemic.However,the detailed molecular immune response at single-cell level is poorly un-derstood.In this study,we systematically delineated the landscape of the pre-and post-vaccination single-cell transcriptome,TCR(T cell antigen receptor)and BCR(B cell antigen re-ceptor)expression profile of vaccinated candidates.The bulk TCR sequencing analysis of COV-ID-19 patients was also performed.Enrichment of a clonal CD8+T cell cluster expressingspecific TCR was identified in both vaccination candidates and COVID-19 patients.These clonalCD8+T cells showed high expression of cytotoxicity,phagosome and antigen presentationrelated genes.The cellecell interaction analysis revealed that monocytes and dendritic cellscould interact with these cells and initiate phagocytosis via ICAM1-ITGAM and ITGB2 signaling.Together,our study systematically deciphered the detailed immunological response duringSARS-CoV-2 vaccination and infection.It may facilitate understanding the immune responseand the T-cell therapy against COVID-19.

Reprogramming Mycobacterium tuberculosis CRISPR System for Gene Editing and Genomewide RNA Interference Screening

Mycobacterium tuberculosis is the causative agent of tuberculosis(TB), which is still the leading cause of mortality from a single infectious disease worldwide. The development of novel anti-TB drugs and vaccines is severely hampered by the complicated and time-consuming genetic manipulation techniques for M. tuberculosis. Here, we harnessed an endogenous type Ⅲ-A CRISPR/Cas10 system of M. tuberculosis for efficient gene editing and RNA interference(RNAi).This simple and easy method only needs to transform a single mini-CRISPR array plasmid, thus avoiding the introduction of exogenous protein and minimizing proteotoxicity. We demonstrated that M. tuberculosis genes can be efficiently and specifically knocked in/out by this system as confirmed by DNA high-throughput sequencing. This system was further applied to single-and multiple-gene RNAi. Moreover, we successfully performed genome-wide RNAi screening to identify M. tuberculosis genes regulating in vitro and intracellular growth. This system can be extensively used for exploring the functional genomics of M. tuberculosis and facilitate the development of novel anti-TB drugs and vaccines.