Antibody Recognition of Human Epidermal Growth Factor Receptor-2 (HER2) Juxtamembrane Domain Enhances Anti-Tumor Response of Chimeric Antigen Receptor (CAR)-T Cells.

Chimeric antigen receptor (CAR) T cell therapy shows promise in treating malignant tumors. However, the use of human epidermal growth factor receptor-2 (HER2) CAR-T cells carries the risk of severe toxicity, including cytokine release syndrome, due to their "on-target off-tumor" recognition of HER2. Enhancing the quality and functionality of HER2 CARs could greatly improve the therapeutic potential of CAR-T cells. In this study, we developed a novel anti-HER2 monoclonal antibody, Ab8, which targets domain III of HER2, distinct from the domain IV recognition of trastuzumab. Although two anti-HER2 mAbs induced similar levels of antibody-dependent cellular cytotoxicity, trastuzumab-based CAR-T cells exhibited potent antitumor activity against HER2-positive cancer cells. In conclusion, our findings provide scientific evidence that antibody recognition of the membrane-proximal domain promotes the anti-tumor response of HER2-specific CAR-T cells.

Serum 25-hydroxyvitamin D is associated with homocysteine in infertile patients with polycystic ovary syndrome (PCOS).

OBJECTIVES: The aim of the study was to investigate whether there is a relationship between serum 25-hydroxyvitamin D and homocysteine in infertile-related PCOS. MATERIAL AND METHODS: We retrospectively reviewed 208 participants (86 PCOS and 122 non-PCOS) who met the inclusion and exclusion criteria from March 2020 to October 2021 at the Department of Obstetrics and Gynecology of the Second affiliated hospital of Xi'an Jiaotong University. Methods of Pearson correlation and linear regression were used to evaluate the associations between serum levels of 25-hydroxyvitamin D and homocysteine in infertile-related PCOS, and a smooth curve fitting were used to address potential nonlinearity. RESULTS: An inverse association between serum 25-hydroxyvitamin D and homocysteine was observed (r = -0.392, p < 0.001) in PCOS groups. Multiple linear regression analysis showed serum 25-hydroxyvitamin D was independently negatively associated with homocysteine levels after controlling for confounding factors (ß = -0.316, p = 0.006). Age, BMI-stratified multivariate linear regression showed that serum 25-hydroxyvitamin D were independently associated with hyperhomocysteine especially in PCOS women aged 30 years or younger after adjusting age, BMI, and AMH. CONCLUSIONS: Herein, the current findings suggest that 25-hydroxyvitamin D levels was negatively associated with serum homocysteine in women with infertility-related PCOS.

Severe major vessel injury during peadicle screw removal: a case report.

Introduction: Pedicle screw fixation (PSF) has been the standard therapy for the treatment of various spinal diseases. Although complications are identified regularly, iatrogenic vascular injury is one of the rare but life-threatening complications. In this literature, we describe the first case of inferior vena cava (IVC) injury during pedicle screw removal. Case description: A 31-year-old man was treated by percutaneous pedicle screw fixation for an L1 compression fracture. After a year, the fracture healed well and hardware removal surgery was performed. During the procedure, the hardware on the right was removed unremarkably except for the L2 pedicle screw which slipped into the retroperitoneum because of the improper technique. The CT angiogram revealed the screw had breached the anterior cortex of the L2 vertebral body and penetrated the IVC. After multidisciplinary cooperation, the defect of IVC was reconstructed and the L2 screw was removed from the posterior approach in the end. Result: The patient recovered well and was discharged after 3 weeks without further events. The removal of the contralateral implants was unremarkable at 7 months postoperatively. At the 3-year follow-up, the patient returned to his normal daily activity without any complaints. Conclusion: Although pedicle screw removal is a rather simple procedure, severe complications may have occurred from this procedure. Surgeons should keep vigilant to avoid the complication noted in this case.

Association between the serum vitamin D level and prevalence of obesity/abdominal obesity in women with infertility: a cross-sectional study of the National Health and Nutrition Examination Survey data.

AIMS: To explore the relationship between vitamin D and obesity and abdominal obesity in women with infertility. MATERIAL AND METHODS: We screened the data from National Health and Nutrition Examination Survey (NHANES) 2013-2016. A total of 201 infertile women between the ages of 20 and 40 years were included in our study. To estimate the independent association of vitamin D with obesity and abdominal obesity, we used weighted multivariate logistic regression models and cubic spline analyses. RESULTS: Among infertile women in the NHANES 2013-2016 database, serum vitamin D levels were significantly and negatively associated with body mass index (ß= -0.96, 95% CI: -1.40, -0.51, p < 0.001)and waist circumference (ß= -0.40, 95% CI: -0.59, -0.22, p < 0.001), respectively. After multivariable adjustment, lower vitamin D levels were found to be associated with a higher prevalence of obesity (OR: 8.290, 95% CI: 2.451-28.039, p for trend = 0.001) and abdominal obesity (OR: 4.820, 95%CI: 1.351-17.194, p for trend =0.037). Spline regression showed linearity of the associations between vitamin D and obesity/abdominal obesity (p for nonlinearity > 0.05). CONCLUSION: Our findings suggested that a decreased vitamin D might correspond to a higher prevalence of obesity in infertile women, which reminded us to pay more attention to the supplement of vitamin D in obese infertile women.

Reliability and Validity of A Novel Device for Evaluating the Cervical Proprioception.

INTRODUCTION: In clinical practice, cervical proprioception is usually evaluated by calculating the cervical joint position error (JPE) with laser pointer devices (LPD) or cervical range-of-motion (CROM) instruments. As technology continues to improve, more and more advanced tools are used to evaluate cervical proprioception. The purpose of this study was to analyze the reliability and validity of the WitMotion sensor (WS) device in evaluating cervical proprioception, and to explore a cheaper, more convenient, and more practical testing tool. METHODS: Twenty-eight healthy participants (16 women, 12 men; age 25-66 years) were recruited and evaluated for cervical joint position error with a WS and LPD by two independent observers. All participants repositioned their head to the target position and the deviation of repositioning was calculated using these two instruments. The intra- and inter-rater reliability of the instrument were determined by calculating the intraclass correlation coefficients (ICC), and the validity was analyzed by calculating the ICC and the Spearman's correlation. RESULTS: The intra-rater reliability of the WS (ICCs = 0.682-0.774) was higher than that of the LPD (ICCs = 0.512-0.719) for measuring JPE of cervical flexion, right lateral flexion, and left rotation. However, the LPD (ICCs = 0.767-0.796) outperformed the WS (ICCs = 0.507-0.661) in cervical extension, left lateral flexion, and right rotation. For the inter-rater reliability, the ICC values obtained by the WS and the LPD were above 0.70 for all cervical movements except cervical extension and left lateral flexion (ICCs = 0.580-0.679). For the validity, the ICC values were moderate to good (ICCs > 0.614) for measuring JPE in all movements with the WS and the LPD. CONCLUSIONS: Based on the high ICC values of reliability and validity, the novel device can be an alternative tool to evaluate cervical proprioception in clinical practice. TRIAL REGISTRATION: This study was registered in the Chinese Clinical Trial Registry (ChiCTR2100047228).

Overexpressed ski efficiently promotes neurorestoration, increases neuronal regeneration, and reduces astrogliosis after traumatic brain injury.

Traumatic brain injury (TBI) survivors suffer from long-term disability and neuropsychiatric sequelae due to irreparable brain tissue destruction. However, there are still few efficient therapies to promote neurorestoration in damaged brain tissue. This study aimed to investigate whether the pro-oncogenic gene ski can promote neurorestoration after TBI. We established a ski-overexpressing experimental TBI mouse model using adenovirus-mediated overexpression through immediate injection after injury. Hematoxylin-eosin staining, MRI-based 3D lesion volume reconstruction, neurobehavioral tests, and analyses of neuronal regeneration and astrogliosis were used to assess neurorestorative efficiency. The effects of ski overexpression on the proliferation of cultured immature neurons and astrocytes were evaluated using imaging flow cytometry. The Ski protein level increased in the perilesional region at 3 days post injury. ski overexpression further elevated Ski protein levels up to 14 days post injury. Lesion volume was attenuated by approximately 36-55% after ski overexpression, with better neurobehavioral recovery, more newborn immature and mature neurons, and less astrogliosis in the perilesional region. Imaging flow cytometry results showed that ski overexpression elevated the proliferation rate of immature neurons and reduced the proliferation rate of astrocytes. These results show that ski can be considered a novel neurorestoration-related gene that effectively promotes neurorestoration, facilitates neuronal regeneration, and reduces astrogliosis after TBI.

Remodelling of tumour microenvironment by microwave ablation potentiates immunotherapy of AXL-specific CAR T cells against non-small cell lung cancer.

The complex immunosuppressive tumour microenvironment (TME) and lack of tumour-specific targets hinder the application of chimeric antigen receptor (CAR) T cells in the treatment of solid tumours. Combining local treatment with CAR T cell immunotherapy may regulate the TME and enhance the killing potency of CAR T cells in solid tumours. Here, we show that AXL, which is highly expressed in non-small cell lung cancer (NSCLC) but not in normal tissues, might be a target for CAR T cell therapy. AXL-CAR T cells alone cause moderate tumour regression in subcutaneous and pulmonary metastatic lung cancer cell-derived xenograft models. Combination of microwave ablation (MWA) and AXL-CAR T cells have superior antitumour efficacy. MWA enhances the activation, infiltration, persistence and tumour suppressive properties of AXL-CAR T cells in AXL-positive NSCLC patient-derived xenograft tumours via TME remodelling. The combination therapy increases the mitochondrial oxidative metabolism of tumour-infiltrating CAR T cells. Combination treatment induces significant tumour suppression without observed toxicities in humanized immunocompetent mice. The synergistic therapeutic effect of MWA and AXL-CAR T cells may be valuable for NSCLC treatment.

Ideal entry point and trajectory for C2 pedicle screw placement in children: a 3D computed tomography study.

PURPOSE: To identify the ideal entry point for pediatric C2 pedicle screw and to obtain parameters of it for the indication of pediatric atlantoaxial fusion arthrodesis. METHODS: The pediatric cervical CT images were reconstructed into the 3D digital models and the C2 vertebrae were separated. The location of ideal entry point and screw placement related linear and angular parameters were assessed on the 3D digital models. RESULTS: A total of 214 pedicles from 107 C2 digital models were analyzed. The average entry point for C2 was 3.80 ± 2.78 mm medial to the lateral notch (LN) and 2.57 ± 1.70 mm superior to the LN. The average pedicle diameter (PD) was 6.02 ± 1.31 mm, and the average pedicle screw length (PSL) was 25.63 ± 3.46 mm. Statistical differences were found between different sex for PD and PSL (P < 0.05). As patient age increases, using the most lateral and inferior edge of the lateral mass as a reference marker, the entry point tends to move medial and cephalad, when using the LN as a reference marker, the entry point tends to move medial and slightly caudad. Univariate linear regression analysis suggested that these linear parameters were associated with age (P < 0.01). CONCLUSION: In this study, we found that the measurement results of C2 pedicle screw varied based on sex, laterality, and ages for children younger than 18 years. The entry point of the screws facilitating ideal trajectory tends to change in a linear way as a function of age. This information helps the surgeon to establish the specific anatomy related to C2 pedicle screw placement to facilitate fixation in the pediatric patients.

Quantitation and Identification of Therapeutic Anti-CD22 Monoclonal Antibodies in a Cell-Based ELISA Method.

Since they lack native soluble membrane antigens, the analysis and selection of antigen-specific antibodies are commonly performed on whole live cells. Here, we have developed a simple and convenient enzyme-linked immunosorbent assay (ELISA) based on cell membrane antigens. Soluble cell membrane proteins isolated from Raji cells were immobilized on the polystyrene microplate, which permitted the assessment of a therapeutic anti-CD22 monoclonal antibody. The experiments showed less variability in the intra-assay. Compared to the living cell ELISAs, the advantage of the assay is avoiding cell losses and high variation of optical density (OD) readings. We provide a quantitative and reproducible ELISA that can be potentially applied to the development of specific antibodies against cell surface antigens.

2022 update on the scientific premise and clinical trials for IL-15 agonists as cancer immunotherapy.

Diverse cytokines and their receptors on immune cells constitute a highly complex network in the immune system. Some therapeutic cytokines and their derivatives have been approved for cancer treatment. IL-15 is an immune-regulating cytokine with multiple functions, among which the function of activating the immunity of cancer patients has great potential in cancer immunotherapy. In this review, we introduce the functions of IL-15 and discuss its role in regulating the immune system in different immune cells. Meanwhile, we will address the applications of IL-15 agonists in cancer immunotherapy and provide prospects for the next generation of therapeutic designs. Although many challenges remain, IL-15 agonists offer a new therapeutic option in the future direction of cancer immunotherapy.

Application of high precision wavelength calibration method for plasmas rotation measurement based on Fabry-Pérot etalon on experimental advanced superconducting tokamak.

Analyzing the radiation spectra of impurity ions is a widely applied diagnostic scheme for plasma ion temperature and rotation measurements on tokamaks. High precision wavelength calibration is a prerequisite for the accurate measurement of plasma parameters, especially for plasma rotation. Furthermore, the sparseness or absence of the standard spectral lines brings calibration challenges due to the narrow wavelength range. A precise wavelength calibration method is demonstrated in which the comb-like spectra generated by the Fabry-Pérot etalon can lock a series of fixed peaks as reference points in a wide wavelength range. The equal frequency intervals of the comb-like spectra are further corrected using several characteristic neon lines of known wavelengths. The experimental results indicate that the wavelength accuracy obtained by this calibration method is less than 0.005 nm, which corresponds to a rotation speed of 2.3 km/s in the toroidal direction for the beam emission spectroscopy spectrometer installed on the experimental advanced superconducting tokamak. Taking the O V(650.024 nm, n = 4 → 3) line as an example, the maximum difference in the oxygen ion rotation velocity is 3.8 km/s for the absolute rotation of ∼25 km/s, when compared with the calibration results of a standard lamp.

Identification of novel FUS and TARDBP gene mutations in Chinese amyotrophic lateral sclerosis patients with HRM analysis.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by progressive loss of motor neurons. More than 30 genes have been linked to ALS to date, including FUS and TARDBP, which exhibit similar roles in RNA metabolism. This study explored the use of high-resolution melting (HRM) analysis to screen for FUS and TARDBP mutation hotspot regions in 146 Chinese ALS patients, which achieved 100% detection. Two FUS mutations were observed in two different familial ALS probands, a missense mutation (p.R521H) and a novel splicing mutation (c.1541+1G>A). Five TARDBP mutations were identified in six ALS patients, including a novel 3'UTR mutation (c.*731A>G) and four missense mutations (p.G294V, p.M337V, p.G348V, and p.I383V). We found that FUS mutations were present in 1.4% of Chinese ALS patients, whereas TARDBP mutations were responsible for 4.1% of Chinese ALS cases. Here, we describe the accuracy of using highly sensitive HRM analysis to identify two novel FUS and TARDBP mutations in Chinese sporadic and familial ALS cases. Our study contributes to the further understanding of the genetic and phenotypic diversity of ALS.

PKC Mediates LPS-Induced IL-1ß Expression and Participates in the Pro-inflammatory Effect of A2AR Under High Glutamate Concentrations in Mouse Microglia.

Pathogens such as bacterial lipopolysaccharide (LPS) play an important role in promoting the production of the inflammatory cytokines interleukin-1 beta (IL-1ß) and tumour necrosis factor-α (TNF-α) in response to infection or damage in microglia. However, whether different signalling pathways regulate these two inflammatory factors remains unclear. The protein kinase C (PKC) family is involved in the regulation of inflammation, and our previous research showed that the activation of the PKC pathway played a key role in the LPS-induced transformation of the adenosine A2A receptor (A2AR) from anti-inflammatory activity to pro-inflammatory activity under high glutamate concentrations. Therefore, in the current study, we investigated the role of PKC in the LPS-induced production of these inflammatory cytokines in mouse primary microglia. GF109203X, a specific PKC inhibitor, inhibited the LPS-induced expression of IL-1ß messenger ribonucleic acid and intracellular protein in a dose-dependent manner. Moreover, 5 µM GF109203X prevented LPS-induced IL-1ß expression but did not significantly affect LPS-induced TNF-α expression. PKC promoted IL-1ß expression by regulating the activity of NF-κB but did not significantly impact the activity of ERK1/2. A2AR activation by CGS21680, an A2AR agonist, facilitated LPS-induced IL-1ß expression through the PKC pathway at high glutamate concentrations but did not significantly affect LPS-induced TNF-α expression. Taken together, these results suggest a new direction for specific intervention with LPS-induced inflammatory factors in response to specific signalling pathways and provide a mechanism for A2AR targeting, especially after brain injury, to influence inflammation by interfering with A2AR.

Circulating anti-mullerian hormone as predictor of ovarian response to clomiphene citrate in women with polycystic ovary syndrome.

BACKGROUND: To investigate the impact of high circulating AMH on the outcome of CC ovulation induction in women with PCOS. METHODS: This prospective cohort observational study included 81 anovulatory women with PCOS who underwent 213 cycles of CC ovarian stimulation. Serum AMH concentrations were measured on cycle day 3 before the commencement of CC in the first cycle, which were compared between responders and CC-resistant anovulation (CRA). Logistic regression analysis was applied to study the value of serum AMH for the prediction of ovarian responsiveness to CC stimulation. The receiver-operating characteristic (ROC) curve was used to evaluate the prognostic value of circulating AMH. MAIN OUTCOME MEASURES: Serum AMH levels. RESULTS: Women who ovulated after CC therapy had a significantly lower AMH compared with the CRA (5.34 ± 1.97 vs.7.81 ± 3.49, P < 0.001). There was a significant gradient increase of serum AMH levels with the increasing dose of CC required to achieve ovulation (P < 0.05). In multivariate logistic regression analysis, AMH was an independent predictor of ovulation induction by CC in PCOS patients. ROC curve analysis showed AMH to be a useful predictor of ovulation induction by CC in PCOS patients, having 92 % specificity and 65 % sensitivity when the threshold AMH concentration was 7.77 ng/ml. CONCLUSION: Serum AMH may be clinically useful to predict which PCOS women are more likely to respond to CC treatment and thus to direct the selection of protocols of ovulation induction.