Construction of Switch Modules for CAR-T Cell Treatment Using a Site-Specific Conjugation System.

Chimeric antigen receptor T-cell (CAR-T cell) therapy has become a promising treatment option for B-cell hematological tumors. However, few optional target antigens and disease relapse due to loss of target antigens limit the broad clinical applicability of CAR-T cells. Here, we conjugated an antibody (Ab) fusion protein, consisting of an Ab domain and a SpyCatcher domain, with the FITC-SpyTag (FITC-ST) peptide to form a bispecific safety switch module using a site-specific conjugation system. We applied the safety switch module to target CD19, PDL1, or Her2-expressing tumor cells by constructing FMC63 (anti-CD19), antiPDL1, or ZHER (anti-Her2)-FITC-ST, respectively. Those switch modules significantly improved the cytotoxic effects of anti-FITC CAR-T cells on tumor cells. Additionally, we obtained the purified CD8+ T cells by optimizing a shorter version of the CD8-binding aptamer to generate anti-FITC CD8-CAR-T cells, which combined with the CD4-FITC-ST switch module (anti-CD4) to eliminate the CD4-positive tumor cells in vitro and in vivo. Overall, we established a novel safety switch module by site-specific conjugation to enhance the antitumor function of universal CAR-T cells, thereby expanding the application scope of CAR-T therapy and improving its safety and efficacy.

Bilateral fronto-orbital advancement combined with cranial vault release using a free-floating bone flap technique for nonsyndromic unilateral coronal synostosis.

Background: The goals of operative treatment for unilateral coronal synostosis (UCS) are to improve appearance and allow unrestricted brain growth. However, for severe unilateral premature closure of the coronal suture, existing methods do not address the compression of the brain or expand the volume of the skull cavity. We report our retrospective experience with bilateral fronto-orbital advancement combined with cranial vault release using a free-floating bone flap (CVR + FFBF) technique and the resulting changes in the anterior cranial vault asymmetry index (ACVAI) and intracranial volume. Methods: Twenty patients with UCS who underwent bilateral fronto-orbital advancement combined with CVR + FFBF technique from April 2014 to May 2019 were included. Surgical efficacy was evaluated by the ACVAI and intracranial volume before the operation, 1 week after the operation, and at the last follow-up (average 19.8 months; range, 12 to 40 months). The measurement data are presented as the mean ± standard deviation and were statistically analyzed by t-test. Results: The ACVAI was 9.07%±3.55% before the operation, 3.56%±3.42% 1 week after the operation, and 3.13%±2.41% at the last follow-up. The ACVAI 1 week after the operation was significantly lower than that before the operation (t=4.827, P<0.001). There was no significant difference between the ACVAI 1 week after the operation and at the last follow-up (t=0.660, P=0.517). The intracranial volume was 1,027.85±112.25 mL in patients before the operation and 1,131.92±161.71 mL in the normal control group, which was a statistically significant difference (t=2.364, P=0.023). The intracranial volume significantly increased 1 week after surgery: 1,081.62±111.10 mL (t=8.703, P<0.001), and this trend continued at the last follow-up (1,386.90±119.30 mL) similarly to the normal control group (1,438.22±89.28 mL). At the last follow-up, there was no significant difference between the two groups (t=1.540, P=0.132). Conclusions: For the treatment of UCS, bilateral fronto-orbital advancement combined with CVR + FFBF technique offers functional and cosmetic outcomes in terms of intracranial volume expansion and fronto-orbital symmetry.

Insight into the uptake, accumulation, and metabolism of the fungicide phenamacril in lettuce (Lactuca sativa L.) and radish (Raphanus sativus L.).

The fungal species Fusarium can cause devastating disease in agricultural crops. Phenamacril is an extremely specific cyanoacrylate fungicide and a strobilurine analog that has excellent efficacy against Fusarium. To date, information on the mechanisms involved in the uptake, accumulation, and metabolism of phenamacril in plants is scarce. In this study, lettuce and radish were chosen as model plants for a comparative analysis of the absorption, accumulation, and metabolic characteristics of phenamacril from a polluted environment. We determined the total amount of phenamacril in the plant-water system by measuring the concentrations in the solution and plant tissues at frequent intervals over the exposure period. Phenamacril was readily taken up by the plant roots with average root concentration factor ranges of 60.8-172.7 and 16.4-26.9 mL/g for lettuce and radish, respectively. However, it showed limited root-to-shoot translocation. The lettuce roots had a 2.8-12.4-fold higher phenamacril content than the shoots; whereas the radish plants demonstrated the opposite, with the shoots having 1.5 to 10.0 times more phenamacril than the roots. By the end of the exposure period, the mass losses from the plant-water systems reached 72.0% and 66.3% for phenamacril in lettuce and radish, respectively, suggesting evidence of phenamacril biotransformation. Further analysis confirmed that phenamacril was metabolized via hydroxylation, hydrolysis of esters, demethylation, and desaturation reactions, and formed multiple transformation products. This study furthers our understanding of the fate of phenamacril when it passes from the environment to plants and provides an important reference for its scientific use and risk assessment.

Occurrence and dietary risk assessment of 37 pesticides in wheat fields in the suburbs of Beijing, China.

The co-occurrence of multiple pesticides in wheat fields adversely affects human health and the environment. Herein, 206 pairs of wheat and soil samples were collected from wheat fields in Beijing, China from 2018 to 2020. One or multiple pesticide residues were detected, and carbendazim (maximum: 38511.5 µg/kg) and tebuconazole (maximum: 45.4 µg/kg) had heavy occurrence in the wheat samples. Carbendazim, triazoles, and neonicotinoids were frequently detected in the soil samples. HCHs and DDTs were detected, with p,p'-DDE in 100.0% of the soil samples at a maximum concentration of 546.0 µg/kg in 2020. Concentrations of carbendazim, tebuconazole, hexaconazole, and cyhalothrin in the paired soil and wheat samples exhibited significant positive correlations. Pesticides that exceeded the maximum residue limits do not pose non-carcinogenic risks, with one exception. The results provide important references towards risk monitoring and control in wheat fields, as well as facilitating the scientific and reasonable use of these pesticides.

Three-dimensional reconstruction and morphological characterization of pituitary macroadenomas.

INTRODUCTION: The aim was to investigate the relationship between the tumor (clinicopathologic and radiological) characteristics and the morphological parameters of pituitary macroadenoma or giant adenoma patients using a three-dimensional (3D) reconstructed model. MATERIAL AND METHODS: Magnetic resoanance imaging (MRI) was performed preoperatively; tumor grade was determined by the Knosp-Steiner classification and tumor morphology by the SIPAP classification. Pituitary adenomas and adjacent structures were reconstructed three-dimensionally by volume rendering. RESULTS: Fifty-two and 6 patients underwent surgery via the transnasal transsphenoidal or pterional approach, respectively. Knosp-Steiner grades I to IV adenomas were observed in 5.2%, 25.9%, 22.4% and 46.6% of the patients, respectively. The 3D model was reconstructed in all cases with superb delineation of tumor morphology and the spatial relationship between the tumor and adjacent tissues. Pituitary adenomas were categorized into intrasellar (13.8%), suprasellar (20.7%), infrasellar (17.2%), and lobulated adenomas (48.3%). Suprasellar adenomas had the smallest (2.27 ±3.22 cm(3)) and lobulated adenomas the largest volume (24.61 ±30.50 cm(3)). Intrasellar adenomas were all functioning, while 75%, 60% and 60.7%, respectively, of suprasellar, infrasellar and lobulated adenomas were nonfunctioning, with a significant association between tumor morphology and secretory function (p = 0.005). CONCLUSIONS: Three-dimensional reconstruction of pituitary macroadenomas offers a simplified morphological classification of pituitary adenomas and may be helpful for neurosurgeons to categorize and characterize pituitary adenomas.

Lateral Craniopharyngeal Canal: An Anatomical Cause of Spontaneous Rhinorrhea.

The aim of this report is to investigate the pathogenesis and surgical treatment of encephalocele located in the sphenoid sinus and presented with spontaneous rhinorrhea. The data of a patient with sphenoid sinus encephalocele was analyzed in association with the review of literature. The patient admitted to our clinic with rhinorrhea from the left nasal cavity. Cranial magnetic resonance imaging revealed bone defects in the left sphenoid sinus wall, and the presence of brain tissue and cerebrospinal fluid (CSF) in the sphenoid sinus. The patient underwent dural repair via left pterional approach. Rhinorrhea disappeared after surgery. The pathogenesis of the encephalocele in the sphenoid sinus is not clear. It may be related to the presence of lateral craniopharyngeal canal, extra-gasification in the sphenoid sinus, high intracranial pressure or other factors. It is very important to diagnose accurately the source of CSF fistula in preoperative period, choose the specific operation technique, and follow-up the patient for a long time period.

Effects of parasagittal meningiomas on intracranial venous circulation assessed by the virtual reality technology.

OBJECTIVE: This study is to investigate the compensatory intracranial venous pathways in parasagittal meningiomas (PSM) patients by virtual reality technology. METHODS: A total of 48 PSM patients (tumor group) and 20 patients with trigeminal neuralgia and hemifacial spasm but without intracranial venous diseases (control group) were enrolled. All patients underwent 3D CE-MRV examination. The 3D reconstructed images by virtual reality technology were used for assessment of diameter and number of intracranial veins, tumor location, venous sinus invasion degree and collateral circulation formation. RESULTS: Diameter of bridging veins in posterior 1/3 superior sagittal sinus (SSS) in tumor group was significantly smaller than that of the control group (P < 0.05). For tumors located in mid 1/3 SSS, diameter of bridging veins and vein of Labbé (VL) in posterior 1/3 SSS decreased significantly (P < 0.05). For tumors located in posterior 1/3 SSS, bridging vein number and transverse sinus (TS) diameter significantly decreased while superficial Sylvian vein (SSV) diameter increased significantly (P < 0.05). Compared with tumor in posterior 1/3 SSS subgroup, number of bridging veins in the tumor in mid 1/3 SSS subgroup increased significantly (P < 0.05). Compared with control group, only the bridging vein number in anterior 1/3 SSS segment in invasion Type 3-4 tumor subgroup decreased significantly (P < 0.05). Diameter of TS and bridging veins in posterior 1/3 SSS segment in sinus invasion Type 5-6 tumor subgroup decreased significantly (P < 0.05). Compared with control group, only the diameter of VL and TS of collateral circulation Grade 1 tumor subgroup decreased significantly (P < 0.05) while in Grade 3 tumor subgroup, TS diameter decreased and SSV diameter increased significantly (P < 0.05). CONCLUSIONS: The intracranial blood flow is mainly drained through SSV drainage after SSS occlusion by PSM.

MRI research of diaphragma sellae in patients with pituitary adenoma.

This study is to investigate the clinical significance of diaphragma sellae in patients with pituitary adenoma by MR images. A total of 47 cases of pituitary adenoma patients were enrolled in this study. Preoperative and postoperative MR scanning together with preoperative 3D-GE sequential scanning were performed. A series of parameters of diaphragma sellae were measured and compared. Tumor height was greater in patients with convex diaphragma sellae than that in patients with concave diaphragma sellae. The width and height of diaphragmal opening were positively related to tumor height. Diaphragmal opening width in the invasive group was greater than that of the non invasive group. Diaphragmal opening width in the non total resection group was significantly greater than that in the total resection group. Tumor resection rate was negatively correlated to diaphragmal opening width in the non total resection group. Lift angle of bilateral epidural around diaphragmal opening was positively related to tumor height. Enhanced 3D-GE images can perfectly display diaphragma sellae and parameters of tumor height and tumor invasiveness are related to diaphragmal opening diameter.

Lentivirus-Mediated knockdown of tectonic family member 1 inhibits medulloblastoma cell proliferation.

Tectonic family member 1 (TCTN1) encodes a member of the tectonic family which are evolutionarily conserved secreted and transmembrane proteins, involving in a diverse variety of developmental processes. It has been demonstrated that tectonics expressed in regions that participate in Hedgehog (Hh) signaling during mouse embryonic development and was imperative for Hh-mediated patterning of the ventral neural tube. However, the expression and regulation of tectonics in human tumor is still not clear. In this study, shRNA-expressing lentivirus was constructed to knockdown TCTN1 in medulloblastoma cell line Daoy. The results showed that knockdown of TCTN1 inhibited cell proliferation and colony formation in Daoy cell line, also caused cell cycle arrest at the G2/M boundary. Taken all together, our data suggest that TCTN1 might play an important role in the progression of medulloblastoma.

Guidance value of intracranial venous circulation evaluation to parasagittal meningioma operation.

OBJECTIVE: This study is to investigate the value of intracranial venous circulation evaluation in guiding the surgery for parasagittal meningioma. METHODS: A total of 44 parasagittal meningioma (PSM) patients received 3D CE-MRV scanning. The obtained images were reconstructed by virtual reality (VR) technology. Venous collateral circulation was evaluated. Postoperative follow-up was carried out. RESULTS: Among 44 PSM patients, 41 cases were with resection of Simpson grade I/II (93.18%) and 6 cases were with permanent neurological dysfunction (13.64%). Venous sinus thrombosis occurred in the remaining 3 patients (6.82%), with 2 cases cured and 1 case died. The mortality rate was 2.27%. Recurrence occurred in one case (2.27%) after discharge. In 9 cases, tumor adjacent SSS was transected and the poor prognosis rate was 33.33%. SSS was opened in 3 cases and after removal of the tumor the broken end was sutured directly. The poor prognosis occurred in one case (33.33%). Electrocoagulation was carried out to the SSS wall in 32 cases without opening venous sinus and 2 cases were with poor prognosis (6.25%). The three different venous sinus treatment methods had no significantly difference on the prognosis of patients (P > 0.05). CONCLUSIONS: Preoperative evaluation of intracranial venous circulation with VR technology and 3D CE-MRV may help making individual surgical plans, reduce venous injury and improve the prognosis of PSM patients.

Effect of small nuclear ribonucleoprotein-associated polypeptide N on the proliferation of medulloblastoma cells.

Spliceosome mutations have been reported in various types of cancer and a number of antitumor drugs have been observed to tightly bind to spliceosome components. Small nuclear ribonucleoprotein­associated polypeptide N (SNRPN) is a small ribonuclear protein and is a key spliceosome constituent. However, the role of SNRPN in human medulloblastoma remains unknown. In the present study, the effect of SNRPN on cell growth was investigated in vitro using the Daoy human medulloblastoma cell line. Lentivirus (Lv)-mediated short hairpin (sh) RNA was used to silence SNRPN expression, which was verified by reverse transcription­quantitative polymerase chain reaction and western blotting. Cell proliferation was examined by MTT and colony formation assays. Knockdown of SNRPN markedly reduced the proliferation and colony formation ability of Daoy medulloblastoma cells. In addition, flow cytometric analysis revealed that the cell cycle distribution was altered when the Daoy cells were infected with Lv­shSNRPN. To the best of our knowledge, this is the first study to investigate the effect of SNRPN on cell proliferation in medulloblastoma. The results indicate that SNRPN may be a potential novel target for the development of pharmacological therapeutics in human medulloblastoma.

Inhibition of GPR137 suppresses proliferation of medulloblastoma cells in vitro.

Medulloblastoma is the most common malignant pediatric brain tumor in children. GPR137 is a ubiquitously expressed gene in the central nervous system. It has been reported that GPR137 modulates malignant proliferation of glioma cells. However, the relationship between GPR137 and medulloblastoma is still unknown. In this study, we knocked down GPR137 in the medulloblastoma cell line Daoy via a lentivirus-based RNA interference system to explore its role in medulloblastoma. Functional analyses showed that cell proliferation and colony formation were obviously restrained in Daoy cells after GPR137 knockdown. Furthermore, knockdown of GPR137 in Daoy cells led to a significant increase in cell percentage in the G0/G1 phase but a decrease in the S phase. Additionally, the cell population in the sub-G1 phase, which represents apoptotic cells, was remarkably increased in GPR137 knockdown cells. GPR137 inhibition induced a strong proapoptotic effect in Daoy cells, as confirmed by annexin V-APC/7-AAD double staining. In conclusion, GPR137 knockdown inhibited growth of Daoy medulloblastoma cells via disturbing cell cycle progression and inducing apoptosis. Our investigation suggested that GPR137 could be a potential oncogene in medulloblastoma cells and might serve as a target for the treatment of medulloblastoma.

A virtual reality model of the clivus and surgical simulation via transoral or transnasal route.

BACKGROUND: Neurosurgery in areas with restricted space and complicated anatomy can be greatly aided by the virtual reality (VR) technique. The clivus represents one of such challenging surgical areas, but its VR has not been established. The present study aimed to document a VR model of clival anatomy that may be useful in clival surgery. METHODS: High resolution CT angiography and MRI were used. The study included a total of 20 patients who did not have any obvious abnormalities detected in the oral, nasal, and clival areas. The images were fused with a Dextroscope. RESULTS: In the VR model, the key structures such as the clival bone, basilar artery, brainstem, pituitary gland, and paranasal sinuses were clearly observed. The morphology of the clivus and its spatial relationships with the neighboring structures were also illustrated. Visualization of the clival model can be made flexible from various planes, angles, or orientations. In addition, surgical access to the clivus via the transoral route or transnasal route was simulated in detail. CONCLUSION: The simulation of the VR model offers a straightforward, three-dimensional, interactive understanding of the size and shape of the clivus, and its relationships with the surrounding blood vessels and bones. It also demonstrates simulated operational procedures such as opening the surgical window, measuring the exposure distance and angles, and determining the critical boundaries in relation to key structures such as the brainstem and arteries. Digitalized VR modeling appears to be helpful for understanding the anatomy of the clivus and its surgical approaches.

Stereoscopic virtual reality models for planning tumor resection in the sellar region.

BACKGROUND: It is difficult for neurosurgeons to perceive the complex three-dimensional anatomical relationships in the sellar region. METHODS: To investigate the value of using a virtual reality system for planning resection of sellar region tumors. The study included 60 patients with sellar tumors. All patients underwent computed tomography angiography, MRI-T1W1, and contrast enhanced MRI-T1W1 image sequence scanning. The CT and MRI scanning data were collected and then imported into a Dextroscope imaging workstation, a virtual reality system that allows structures to be viewed stereoscopically. During preoperative assessment, typical images for each patient were chosen and printed out for use by the surgeons as references during surgery. RESULTS: All sellar tumor models clearly displayed bone, the internal carotid artery, circle of Willis and its branches, the optic nerve and chiasm, ventricular system, tumor, brain, soft tissue and adjacent structures. Depending on the location of the tumors, we simulated the transmononasal sphenoid sinus approach, transpterional approach, and other approaches. Eleven surgeons who used virtual reality models completed a survey questionnaire. Nine of the participants said that the virtual reality images were superior to other images but that other images needed to be used in combination with the virtual reality images. CONCLUSIONS: The three-dimensional virtual reality models were helpful for individualized planning of surgery in the sellar region. Virtual reality appears to be promising as a valuable tool for sellar region surgery in the future.

Diagnostic Significance of Intracystic Nodules on MRI in Rathke's Cleft Cyst.

Background and Purpose. To explore strategies for the diagnosis and treatment of Rathke's cleft cyst (RCC). Methods. The medical records of 24 patients with sellar RCC were retrospectively reviewed. Two patients had concomitant pituitary adenoma, 2 underwent transcranial surgery, and 22 underwent transsphenoidal surgery. The clinical features, especially the findings of intracystic nodules on MRI, were evaluated and compared with the pathological findings. Results. Preoperatively, only 2 patients were diagnosed with RCC or suspected RCC. Pre- and postoperative MRI images revealed 10 intracystic nodules in 9 (37.5%) patients. Two nodules had bull's eyelike changes. The signal intensity of the intracystic nodules varied on T1- and T2-weighted images. Not all nodules on T2-weighted images were visualized. Postoperative MRI revealed recurrence or residual lesion in 5 patients; none had new symptoms and a second surgery was not required. Conclusions. Identifying intracystic nodules is important in patients with sellar cystic lesions. Bull's eyelike change in an intracystic nodule on MRI, which is reported here for the first time, potentially might have value for confirming the diagnosis.

The effect of galectin-3 genetic variants on the susceptibility and prognosis of gliomas in a Chinese population.

The aim of this study is to explore the association between the polymorphisms of galectin-3 gene and clinico-pathological characteristics and prognosis of gliomas. We enrolled 190 histologically diagnosed gliomas and 210 healthy controls in this study. Two genetic variants at galectin-3 single nucleotide polymorphism (SNP) sites (galectin-3 +191 A>C and +292 A>C) were determined. We found that the A/A genotype at galectin-3 gene +292 A>C was significantly more prevalent in gliomas patient than in controls (42.1% vs. 29.0%, P=0.021); the A allele frequency was markedly higher in gliomas subjects than in controls (61.8% vs. 45.0%, P=0.008). There was a markedly higher prevalence of AA carriers in high-grade subgroup than in low-grade subgroup (50.5% vs. 31.8%, P=0.012). The Kaplan-Meier analyses showed that the gliomas patients carrying AA genotype of galectin-3 gene +292 A>C had marked shorter overall survival period than those did not (AA vs. AC+CC, 22.2±3.8 months vs. 38.3 months±7.9; P=0.04). The SNPs at +191 A>C of galectin-3 gene did not show positive association with clinico-pathological characteristics and prognosis of gliomas. The results of this study suggest the SNPs at +292 A>C, not SNPs at +191 A>C, of galectin-3 gene were associated with the tumor grade and prognosis of gliomas.