KRAS mutation represents a formidable challenge in cancer treatment. Here, we explored the CasRx-editing approach on lung adenocarcinoma driven by KRAS activation and Tp53 loss in vivo, a scenario closely resembling clinical conditions. We screened seven sgRNAs in vitro and identified CasRx + sgRNA5 effectively suppressed Kras-G12D transcript levels, leading to attenuation of cell growth. Subsequently, we administered CasRx + sgRNA5 via in vivo Adeno-associated viruses (AAV) delivery at the early tumorigenesis stage, resulting in inhibition of tumor initiation and progression, ultimately enhancing the survival of the mice. Mechanistically, the treatment directly inhibited KRAS signal pathways and regulated a potential protein network comprising KRAS, DUSP8, NR4A1, DUSP1, and EGR1. Encouragingly, the CasRx + sgRNA5 treatment in mice bearing the induced adenocarcinoma demonstrated a positive therapeutic outcome, characterized by seemingly reduced mutation burden, diminished tumor size, and trend of improved overall survival. Remarkably, no obvious off-target events were detected, highlighting the specificity and safety of CasRx therapy. This study establishes the feasibility of CasRx-based-Kras-G12D editing for in vivo tumor inhibition and provides insights into the potential role of the KRAS-associated protein network in early development of lung cancer. These findings hold promise for development of CasRx therapies for this aggressive malignancy.

Lung cancer is the leading cause of cancer-related deaths worldwide, synthesizing and screening of novel anti-cancer drugs provides an alternative therapeutic strategy for renewal of the chemotherapy regimens against lung cancer. To this end, several compounds were synthesized based on the modification of the original myricetin, and their anti-tumor activity against the human non-small cell lung cancer (NSCLC) A549 cells were measured. Among the myricetin derivatives, S4-10 has displayed the highest antitumor efficacy in dose-dependent manner. The proliferation of A549 cells were significantly attenuated by given 6 µM of S4-10 both in vitro and in vivo. Further, the treatment of S4-10 also results in the inhibition of cell migration and invasiveness and the induction of cell apoptosis and G2 cycle arrest of A549 cells. Moreover, we found that S4-10 inhibits the progression of A549 cells through the sterol biosynthetic-cell apoptosis axis. These findings shed the light of developing S4-10 as a promising treatment agent for NSCLC.

The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has resulted in catastrophic damage worldwide. Accordingly, the development of powerful, safe, easily accessible vaccines with long-term effectiveness is understood as an urgently needed countermeasure against this ongoing pandemic. Guided by this strong promise of using AAVs, we here designed, optimized, and developed an AAV-based vaccines (including AAV-RBD(max), AAV-RBD(wt), AAV-2xRBD, and AAV-3xRBD) that elicit strong immune responses against the RBD domain of the SARS-CoV-2 S protein. These immunogenic responses have proven long-lived, with near peak levels for at least six months in mice. Notably, the sera immunized with AAV-3xRBD vaccine contains powerful neutralizing antibodies against the SARS-CoV-2 pseudovirus. Further evidence proven that potent specific antibodies could also be elicited in canines after vaccination with AAV-3xRBD vaccine.

Brachial plexus (BP) root injury often results in disability of the upper extremities. Improvements in high-frequency ultrasonography have enabled the visualization of BP nerve roots. This study was aimed at quantifying the diagnostic accuracy of ultrasonography in BP root injury at different stages post-trauma. A consecutive series of 170 patients with BP root injury between 2015 and 2019 were studied retrospectively and divided into three groups on the basis of time between injury and ultrasound examination (<= 1 mo, 1-3 mo, >3 mo). Diagnosis of complete BP root injury under ultrasound was determined using a pre-defined criterion, including pseudomeningocele, retraction and rupture. Diagnostic accuracy was calculated based on surgical findings and intra-operative electrophysiological tests. Rates of detection of the cervical (C5-C8) and thoracic (T1) nerve roots under ultrasound were 99.4%, 99.4%, 99.4%, 95.9% and 79.4%, respectively. The sensitivity for complete BP root injury was 0.74, and the specificity was 0.91. No significant differences in sensitivity or specificity were observed across time stages. Ultrasound exhibited substantial consistency with surgical findings (k = 0.70) for complete BP root injury at any stage post-injury. Ultrasound can be an optional method of diagnosis of complete BP root injury at an early stage post-injury. (C) 2022 World Federation for Ultrasound in Medicine & Biology. All rights reserved.

The incidence of hand diseases is increasing.The most common hand dysfunction is the stiffness of the second to fifth metacarpophalangeal joints.We designed a hand functional rehabilitation therapy instrument for passive metacarpophalangeal joint exercise.The instrument was controlled by parameters such as joint motion times.Monte-Carlo method was used to design the hand rehabilitation therapy instrument parameters.K-means clustering algorithm was used to cluster the patient's charac-teristics for parameters collection.Initial parameter values were then assigned to new patients according to the cluster analysis results.

Background . Pancreatic cancer is associated with high death rates and limited therapeutic options, with no effective predictive factors being available for prognosis at present. In this study, we evaluate the value of using blood test results for pancreatic cancer prognosis. Method . The records of 214 pancreatic cancer patients were reviewed. Blood test results for white blood cell (WBC), granulocyte, neutrophil, lymphocyte, platelet count, neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR) were dichotomized on the basis of median values. This was followed by univariate and multivariate analyses between groups. Results . Patients with pretreatment values in the rangeWBC≥5.8×109/L,granulocyte≥3.7×109/L,neutrophil≥3.9×109/L,lymphocyte<1.4×109/L, andNLR≥2.8showed significant correlations pointing to poorer overall survival. Multivariate analysis indicated thatWBC≥5.8×109/L(HR=1.808; 95%CI=1.055–3.096;p=0.031) andgranulocyte≥3.7×109/L(HR=7.346; 95% CI = 1.275–42.321;p=0.026) can be taken to be independent prognostic factors for overall survival in pancreatic patients. Conclusion . Pretreatment values of WBC and granulocyte count were independent factors with poor prognosis ability with respect to pancreatic cancer.

PURPOSE:To identify the pretreatment values of bilirubin and albumin and other serum biomarkers in predicting the prognosis for advanced pancreatic cancer. METHODS:A total of 201 consecutive patients pathologically diagnosed as advanced pancreatic cancer were retrospectively reviewed. Tumor location, TNM classification, the level of baseline total bilirubin (TBIT), direct bilirubin (DBIT), albumin (ALB), globulin (GLOB), total protein (TP), ALB to GLOB ratio (AGR), CA19-9, CA242, and CA50 were collected. The values of CA19-9, CA242, and CA50 were divided into two groups according to the upper limit value which were 1000 U/mL, 150 IU/mL, and 500 IU/mL, respectively. The values of TBIL, DBIL, IBIL, ALB, GLOB, TP, ALB, and GLOB were divided into low and high groups according to the median. To investigate if the median was an effective discriminator in dividing these markers, the patients were divided into a test set (n = 100) and a validation set (n = 101). Kaplan-Meier (K-M) survival analysis and Cox regression analysis were performed to explore the potential relationship between them and overall survival (OS). RESULTS:A K-M survival analysis revealed that the investigated markers in test set, including TBIL, DBIL, IBIL, ALB, GLOB, TP, ALB, and GLOB, were not associated with the OS. The findings from the validation set were consistent with those in the test set. Factors with P value smaller than 0.1 in the univariate analysis along with the tumor location, CA19-9, CA242, CA50, were entered into the multivariate analysis. A Cox regression analysis suggested that the cancerization at head of pancreas (P = 0.01) and a high level of CA19-9 (P = 0.02) were independent prognostic indicators for poor OS of pancreatic cancer. CONCLUSIONS:Baseline bilirubin and serum proteins were not associated with the prognosis of advanced pancreatic cancer. Tumor location and level of CA19-9 may serve as significant indicators for poor prognosis in those patients.

This study aimed to observe the morphological characteristics of a PGLA [poly(glycolide-co-L-lactide)] nerve conduit and regenerated nerve bundle in the human body using high-frequency ultrasound and examine functional recovery of the regenerated nerve using functional magnetic resonance imaging (fMRI) after neural prosthesis with a PGLA nerve conduit. Thirty-nine patients underwent high-frequency ultrasound, and one patient with superficial radial nerve injury (27-mm defect) underwent fMRI at one, three, and six postoperative months. The fMRI examination results were compared with sensory detection and high-frequency ultrasound results during the same follow-up window period. The normal and regenerated nerve bundles had similar ultrasonic imaging features. At one postoperative month, fMRI displayed activeness of the normal cortex in the brain region corresponding to the contralateral superficial radial nerve, while no activeness was observed on the ipsilateral side. From three to six postoperative months, fMRI revealed gradually increasing activeness in the brain region corresponding to the ipsilateral superficial radial nerve, but the activation area on the ipsilateral side was smaller than that on the contralateral side. Combining morphological detection of the regenerated nerve using high-frequency ultrasound and functional detection of the regenerated nerve using fMRI may be a valuable method for evaluating repair of peripheral nerve injury.

Objective To explore the compensatory mechanisms after severing the C7 nerve root by studying the morphological changes of motor end-plate in target muscles.Methods Forty-two male SD rats were randomly divided into 7 groups,with 6 each.C7 nerve root on the right side was transected while that on the left side was left intact.At 1,2,4,6,8,and 12 weeks after C7 nerve root transection,the triceps brachii,latissimus dorsi and extensor digitorum communis muscles of the experimental side and the normal side were harvested and subjected to acetylcholinesterase staining,silver-acetylcholinesterase staining,immunofluorescence staining and electron microscopy.The morphological changes of the motor end-plate in these muscles were recorded and analyzed.SPSS 17.0 software was used to calculate the mean,standard deviation and analysis of variance.Results Within 1 to 3 weeks of C7 transection,there were no significant morphological changes of the motor end-plate in target muscles.Between 4 to 8 weeks post transection,a small number of abnormal motor end-plates with features of larger area,lighter acetylcholinesterase staining and severe ultrastructural degeneration under electron microscopy emerged sparsely in some spots in the target muscles.The morphology of motor endplate returned to normal status 12 weeks following C7 transection.Multiple small specialized postsynaptic membrane aggregations appeared in the neighborhood of the original nerve terminal to form new motor end-plates.Conclusion Morphological changes of the motor end-plate happened in target muscles after C7 nerve root transection.These changes returned to normal in 12 weeks.Axonal sprouting derived from adjacent healthy nerve fibers could be the main mechanism of compensation in C7 nerve transfer.

Objective To study the method and effect of using self-made facile fixative bracket combined with Kirschner wire in the treatment of comminuted middle and proximal phalanx fractures. Methods From June 2009 to August 2013, we had treated 26 patients with comminuted middle and proximal phalanx fractures (32 fingers) using self-made facile fixative bracket combined with Kirschner wire and absorbable suture, and all these cases were followed-up and observed regularly. Results All these cases achieved bone healing without osteomyelitis, malunion and fracture displacement after operation, external bone callus formation were found in 9 cases during bone healing process. We evaluated all the cases according to the Tatol Active Motion (TAM) rating system and Tatol Active Flexion (TAF) rating system, excellent cases were 5 (5 lesions), good were 16 (17 lesions), fair in 3 (6 lesions), poor in 2 (4 lesions), overall excellent and good rate were 80.8% (excellent rate of fracture part was 68. 8%). Conclusion Using self-made facile fixative bracket combined with limited internal fixation can provide an effective immobilization, less invasion and early exercises to restore the function of the fingers for comminuted fracture of proximal and middle phalanx.

Injury to peripheral nerves always results in progressive skeletal muscle atrophy and poor functional recovery. Previous studies have demonstrated that transplanting neural stem cells (NSCs) into peripheral nerve can differentiate into neurons and delay muscle atrophy. However, the mechanism was not very clear. In this study, we transplanted the fetal NSCs to the injured nerve and examined new formed neuromuscular junctions (NMJs) in the denervated muscle and arrest of muscle atrophy. In our study, two pregnant Fischer rats were used to harvest fetal NSCs, 70 rats were randomly divided into NSC-transplanted and control groups, five rats without surgery were used as the normal control. A volume of 5 microl culture media with or without fetal NSCs (5 x 10(6)) were transplanted into distal tibial nerve stump after the nerve was transected in two groups, respectively. Three, five, and seven months after denervation, the dry weight of gastrocnemius muscle was found significant heavier, and the fiber area was more retained in NSC-transplanted group comparing to the control group (P < 0.05). Neurons were found in the distal tibial nerves even 7 months after fetal NSCs transplantation. Newly formed NMJs were detected by immunohistochemistry. In addition, the results of electrophysiological analysis and retrograde tracing manifested that the neural pathway between muscle and differentiated neurons was integrity. In conclusions, our study demonstrated that fetal NSCs transplanted into peripheral nerves could differentiate into neurons and form functional NMJs with denervated muscle, which may be beneficial for the treatment of muscle atrophy after peripheral nerve injury.

Objective To study the range and change of stress in normal radiocarpal joint. Methods Eight fresh-frozen forearm specimens were used in this investigation.The specimens were fixed in a special jig.The load of 150 N was distributed to tendons passing through the wrist in a certain proportion.Placed and maintained the wrist in different positions and inserted encapsulated prescale film into the radiocarpal joint,the messages-including the stress,contact area and mean pressure in the film-were analyzed by Photoshop 7.0 and Image J. Results The stress,contact area and mean pressure in the scaphoid and lunate fossa were different and changed according to the position of the wrist. Conclusions The stress,contact area and mean pressure in the scaphoid and lunate fossa are different and will change according to the position of the wrist,which have important physiological functions,including protect the articular cartilage and adapt to the hand motion.

Objective To study the effect and mechanism of transplanting spinal fetal neural stem cells (NSCs) into the peripheral nerve for delaying muscle denervation atrophy. Methods Spinal. fetal NSCs were separated from spinal cord of enceinte 10 to 12 days SD rats, cultured and purified. After three passages, the formed NSC spheres were blew into single cell suspension ( 106/μl×5 μl) and transplanted into the distal part of the transected tibial nerve. 5 μl of cell culture medium was injected into the distal tibial nerve in the control group. Three and 5 months after the transplantation, the distal part of the tibial nerve and the triceps suraes were harvested and identified with specific markers, by means of indirect immunofluorescent staining to evaluate survival and differentiation of transplanted NSCs in the nerve, and to observe the neuromuscular junctions.Results Compare to the control group, atrophy of the triceps suraes muscle was less severe 3 and 5 months after NSCs transplantation. Postsynaptic membrane was also better preserved in NSCs transplanted group. Five months after NSCs transplantation, new synapses (neuromuscular junction) formed in the denervated muscle.Conclusion NSCs transplantation can delay atrophy denervated muscles. NSCs transplantation can not only maintain the structure of postsynaptic membrane, but also form new synapse with the denervated muscle.

Objective To compare the stress,contact area and pressure change between pre-and post four-comer atthrodesls.To study the effect of four-comer arthrodesis to the stress in raaiocarpal joint.Methods Eight fresh-frozen forearm specimens were used in this investigation.The specimen were placed in a specialjig,150 N load was distributed to tendons passing through the wrist according to a certain proportion.Place and maintain the wrist in different positions,the prescale film was encapsulated and inserted into the radiocarpal joint.Then performed a simulated four-comer arthrodesis.did the same process and gained the corresponding results.Compare the three indexes between pre-and post four-comer arthrodesis.Results In norlnal radioearpal joint,the contact areais 3%to 43%in scaphoid fossa and 6%to 30%inlunatefossa,afterfour-comer arthrodesis,it change to 0%to 30%in seaphoid fossa and 7%to 32%in lunate fossa,all the three indexes were changed according to the position of the wrist.In the normal joint,the stress change mild and gradually,post four-comer arthrodesis,the stress change is intense and rigid.Conclusion The stress,contact area and pressure in the seaphoid and lunate fossa are different and change according to the position of the wrist.The change in the maiocarpal joint after four-comer arthrodesis is acute and hard,which will perhaps interfere the cartilage in the long run.

OBJECTIVE:To recover the loss of the shoulder and elbow function after superior trunks injury of brachial plexus through multiple nerves branch transfer simultaneously near the nerve entering points of recipient nerves.METHODS:Four male patients (aged 21-39 years) with superior trunks injury of brachial plexus were treated from February to September 2007. All cases were injured in the traffic accident, left side in 1 case and right side in 3 cases, resulting in the loss of shoulder abduction, shoulder extorsion, shoulder lift and elbow flexion, and the increase of muscle strength of shoulder shrug, elbow extension and finger flexion to above or equal to 4th grade. Patients were hospitalized 3-11 months after injury. Electromyography showed that the functions of accessory nerve, ulnar nerve and the branch to long head of triceps brachii were good, but the function of median nerve was injured partially. The following multiple donor nerves transfer were performed under general anaesthesia, namely from posterior approach accessory nerve to suprascapular nerve, from triceps to axillary nerve, from the partial branch of ulnar nerve to the biceps and/or brachial is muscular branch of musculocutaneous nerve.RESULTS:All incisions healed by first intention. One case suffered postoperative numbness on the ulnar side of hand and was symptomatically relieved after expectant treatment, while 3 cases had no manifestation of the motor and sensory functional injury related to donor nerve. All patients were followed up for 7-12 months. All patients regained the shoulder abduction and the elbow flexion 3-4 months after operation and electromyography showed that there was the regenerative potential in 3 recipient muscles. The shoulder abduction, elbow flexion and the muscle strength of the patients was 30-65 degrees, 90-120 degrees and 3-4 grade, respectively, 6-7 months after operation. Twelve months after operation, the first patient's shoulder abduction, external rotation, superinduction and elbow flexion almost returned to normal, and his shape of triangular muscle and biceps muscle were nearly normal.CONCLUSION:Adopting donor nerves with similar functions to conduct the multiple donor nerves transfer in cord level has the advantages of little functional loss at the donor sites, and fast and sound functional recovery at the recipient sites. It is especially suitable for the superior trunks injury patient with delayed treatment and for the patient with the great risk in supraclavicular exploration, providing a new approach for treating superior trunks injury of brachial plexus.

Objective To compare and evaluate the differences of biomechanical characteristics of four internal fixation used in proximal interphalangeal joint reconstruction,for the purpose of proposing the experimental basis for clinical application.MethodsForty-eight paired specimens of fingers and toes obtained from the embalmed adult cadaver were randomizedly divided into four groups.The joints were fixed in each group with crossed K-wires,the cruciate wire combined with the inclining K-wire,the dorsal mini-plate,and the lateral mini-plate.Biomechanical characteristics were observed after mimic proximal interphalangeal joints movement.ResultsThe dorsal mini-plate and cruciate wires combined with the inclining K-wire provided more sufficient stability while the lateral mini-plate and crossed K-wires provided poor stability.ConclusionThe cruciate wire combined with the inclining K-wire is an optimal internal fixation method.It avoids undesirable soft tissue dissection and complicated manipulation,provides sufficient stability and is suitable for early rehabilitation.

OBJECTIVE To explore the effect of the collateral ligaments and the plantar plate on the flexion of the metatarsophalangeal (MP) joints. METHODS Twenty-four preserved human No. 2-4 digits were obtained from embalmed cadaver feet, which were divided into 2 groups at random. In group A, the bilateral collateral ligaments were cut first, and then the plantar plate was sectioned. They were cut in opposite sequence in group B. Angle of the flexion of MP joint was observed in the same load after the bilateral collateral ligaments and the plantar plate were sectioned in different sequence. From 1994 to 2000, 11 cases were used with this technic, including plantar section in 2 cases and both plantar section and bilateral collateral ligaments cut in 9 cases. RESULTS The angle of flexion of the MP joint before operation in group A is 37.30 +/- 5.42 degrees, it increased 11.29 +/- 2.36 degrees and to 48.60 +/- 2.98 degrees when the bilateral collateral ligaments were cut, and there was significant difference. Later the cut of the plantar plate increased another 5.30 +/- 1.59 degrees and to 53.35 +/- 2.76 degrees. Both have an increasing trend for the angle of flexion of the MP joint (P < 0.01). While in group B, the angle of flexion of the MP joint before operation is 34.59 +/- 5.32 degrees, it increased 6.29 +/- 2.98 degrees and to 40.89 +/- 2.36 degrees when the plantar plate were cut, later the cut of the bilateral collateral ligaments increased another 9.71 +/- 1.94 degrees and to 50.60 +/- 2.01 degrees. Both had an increasing trend for the angle of flexion of the MP joint (P < 0.01). The bilateral collateral ligaments had more influence than the plantar plate (P < 0.01). There was the same effect in different sequence (P > 0.05). In 2 cases with plantar section, the flexion angle of MP joint could achieve 15 degrees to 45 degrees in 2 months. The other 9 cases with both plantar section and bilateral collateral ligaments cut, the MP joint flexion achieved 10.3 degrees to 58.4 degrees in 26.3 months. CONCLUSION The flexion angle of the MP joint can be increased by cutting the bilateral collateral ligaments and the plantar plate.