Dual targeting chimeric antigen receptor cells enhance antitumour activity by overcoming T cell exhaustion in pancreatic cancer.

BACKGROUND AND PURPOSE: Although our previous data indicated that claudin 18 isoform 2 (CLDN18.2)-targeted chimeric antigen receptor (CAR) T cells displayed remarkable clinical efficacy in CLDN18.2-positive gastric cancer, their efficacy is limited in pancreatic ductal adenocarcinoma (PDAC). The tumour microenvironment (TME) is one of the main obstacles to the efficacy of CAR-T and remodelling the TME may be a possible way to overcome this obstacle. The TME of PDAC is characterized by abundant cancer-related fibroblasts (CAFs), which hinder the infiltration and function of CLDN18.2-targeted CAR-T cells. The expression of fibroblast activation protein alpha (FAP) is an important feature of active CAFs, providing potential targets for eliminating CAFs. EXPERIMENTAL APPROACH: In this study, we generated 10 FAP/CLDN 18.2 dual-targeted CAR-T cells and evaluated their anti-tumour ability in vitro and in vivo. KEY RESULTS: Compared with conventional CAR-T cells, some dual-targeted CAR-T cells showed improved therapeutic effects in mouse pancreatic cancers. Further, dual-targeted CAR-T cells with better anti-tumour effect could suppress the recruitment of myeloid-derived suppressor cells (MDSCs) to improve the immunosuppressive TME, which contributes to the survival of CD8+ T cells. Moreover, dual-targeted CAR-T cells reduced the exhaustion of T cells in transforming TGF-ß dependent manner. CONCLUSION AND IMPLICATIONS: The dual-targeted CAR-T cells obtained enhancement of T effector function, inhibition of T cell exhaustion, and improvement of tumour microenvironment. Our findings provide a theoretical rationale for dual-targeted FAP/CLDN 18.2 CAR-T cells therapy in PDAC.

Closed Bipolar Nanoelectrode Array for Ultra-Sensitive Detection of Alkaline Phosphatase and Two-Dimensional Imaging of Epidermal Growth Factor Receptors.

The combination of closed bipolar electrodes (cBPE) with electrochemiluminescence (ECL) imaging has demonstrated remarkable capabilities in the field of bioanalysis. Here, we established a cBPE-ECL platform for ultrasensitive detection of alkaline phosphatase (ALP) and two-dimensional imaging of epidermal growth factor receptor (EGFR). This cBPE-ECL system consists of a high-density gold nanowire array in anodic aluminum oxide (AAO) membrane as the cBPE coupled with ECL of highly luminescent cadmium selenide quantum dots (CdSe QDs) luminophores to achieve cathodic electro-optical conversion. When an enzyme-catalyzed amplification effect of ALP with 4-aminophenyl phosphate monosodium salt hydrate (p-APP) as the substrate and 4-aminophenol (p-AP) as the electroactive probe is introduced, a significant improvement of sensing sensitivity with a detection limit as low as 0.5 fM for ALP on the cBPE-ECL platform can be obtained. In addition, the cBPE-ECL sensing system can also be used to detect cancer cells with an impressive detection limit of 50 cells/mL by labeling ALP onto the EGFR protein on A431 human epidermal cancer cell membranes. Thus, two-dimensional (2D) imaging of the EGFR proteins on the cell surface can be achieved, demonstrating that the established cBPE-ECL sensing system is of high resolution for spatiotemporal cell imaging.

Reverse traction with Kirschner wires and bilateral external fixation device combined with minimally invasive plate oseoynthesis technique for tibial plateau fractures of type Schatzker V and VI.

PURPOSE: Complex bicondylar tibial plateau fracture (TPF) has always been a tricky problem for surgeons. We created a novel external device used intraoperatively consisting of Kirschner wires, and combined with minimally invasive plate oseoynthesis (MIPO) technique to treat complex bicondylar TPFs, and the clinical effect and feasibility were further evaluated. METHODS: From March 2016 to February 2021, 49 cases (29 males and 20 females) were identified as bicondylar TPF, the mean age 47.2 (27-69). All patients adopted the device and MIPO technique. A series of score, complications, and radiographs in the follow-up period, from three months, six months, one year, and two years and the last follow-up, were recorded, from visual analogue score (VAS), hospital for special surgery (HSS), and Short-Form 36 (SF-36), containing physical (PCS) and mental (MCS), and Rasmussen score. RESULTS: Forty-seven patients showed good functional recovery. No patients were lost, mean follow-up time was 28.17 ± 2.81 (24.2-35.4) months. Operation time was 89.80 ± 13.46 (58-110) min. At the last follow-up, VAS was 1.3 ± 0.92 (0-4), HHS was 93.10 ± 2.63 (89-99), PCS was 49.20 ± 7.40 (38-65), and MCS was 50.08 ± 4.77 (43-62). Complications were as follows: cutaneous necrosis (3, 6%), asymptomatic arthritis (3, 6%), symptomatic arthritis (1, 2%), and deep venous thrombosis (1, 2%). Mean fracture healing time was 11.82 ± 1.5 (10-15.4) weeks. All patients got recovery without extra surgery and removed the implants at 12.85 ± 0.76 (11.2-15.4) months. CONCLUSION: Temporary traction device of bilateral external fixator combined with MIPO technique was simple and convenient, with a smaller soft-tissue damage, an easier operational approach, and its worth being promoted.

Mid-Term Follow-Up Results After Hemiarthroplasty Using Long Femoral Stem Prosthesis (Peerless-160) for Intertrochanteric Fractures in Octogenarians.

Purpose: To analyze the clinical effect and imaging data of cementless bipolar hemiarthroplasty employing a long femoral stem (peerless-160) and 2 reconstructed femoral titanium wires to repair intertrochanteric fractures among the octogenarians. Methods: Between June 2014 and August 2016, 58 octogenarians with femoral intertrochanteric fractures underwent the long femoral stem (peerless-160) cementless bipolar hemiarthroplasty by the same surgeon. We assessed clinical and radiological outcomes such as operative time, bleeding volume, blood transfusion volume, length of hospital stay, full weight-bearing walking time, walking ability calculated by Koval classification and Harris Hip Score (HHS), including fracture healing and greater trochanter fragments subsidence. Result: The surgery was successfully performed in all patients. The average operation time was 72.8 ± 13.2 min, the average blood loss during surgery was 225.0 ± 91.4 mL, 200 mL of blood was transfused, the mean duration of hospitalization was 11.9 ± 4.0 days, the mean time of full weight bearing was 12.5 ± 3.8 days. Patients were followed up for 24-68 months, averagely 49.4 ± 10.3 months. During follow-up, 4 (6.9%) patients died, and 1 (1.7%) was completely lost to ask about the recent situation. The average Harris Hip Score at the last follow-up was 87.8 ± 6.1, most of the patients recovered walking ability, under radiological examination, the prosthesis showed no signs of loosening. All trochanteric fractures gradually healed, the clinical and radiographic signs of healing occurred at average of 4.0 ± 1.1 months postoperatively. Conclusion: For osteoporotic unstable intertrochanteric fractures in octogenarians, this study confirmed that the Cementless Bipolar Hemiarthroplasty Using a Long Femoral stem (peerless-160) with double cross binding technique is a satisfactory and safe choice for the octogenarians.

Expressing IL-15/IL-18 and CXCR2 improve infiltration and survival of EGFRvIII-targeting CAR-T cells in breast cancer.

Previously, we have generated EGFRvIII-targeting CAR-T cells and brought hope for treating advanced breast cancer. However, EGFRvIII-targeting CAR-T cells were defined limited anti-tumor efficacy, which might be due to reduced accumulation, persistence of therapeutic T cells in tumor site of breast cancer. CXCLs were highly expressed in tumor environment of breast cancer and CXCR2 is the main receptor for CXCLs. Here, CXCR2 could significantly improve the trafficking and tumor specific accumulation of CAR-T cells both in vivo and in vitro. However, the anti-tumor effect of CXCR2 CAR-T cells were weaken which might be results of the apoptosis of T cells. Cytokines could stimulate Tcell proliferation, such as interleukin (IL)-15 and IL-18. Then, we generated CXCR2 CAR with synthetic IL-15 or IL-18 production. Co-expressing IL-15 or IL-18 could significantly suppress the exhaustion and apoptosis of T cells and enhanced the anti-tumor activity of CXCR2 CAR-T cells in vivo. Further, coexpression IL-15 or IL-18 in CXCR2 CAR-T cells did not cause toxicity. These findings provide a potential therapy strategy of co-expression IL-15 or IL-18 in CXCR2 CAR-T cells for the treatment of advancing breast cancer in the future.

Correlation of CT-derived pectoralis muscle status and COVID-19 induced lung injury in elderly patients.

OBJECTIVES: To explore the association between CT-derived pectoralis muscle index (PMI) and COVID-19 induced lung injury. METHODS: We enrolled 116 elderly COVID-19 patients linked to the COVID-19 outbreak in Nanjing Lukou international airport. We extracted three sessions of their CT data, including one upon admission (T1), one during the first 2 weeks when lung injury peaked (T2) and one on day 14 ± 2 (T3). Lung injury was assessed by CT severity score (CTSS) and pulmonary opacity score (POS). Pneumonia evolution was evaluated by changes of CT scores at T2 from T1(Δ). RESULTS: The maximum CT scores in low PMI patients were higher than those of normal PMI patients, including CTSS1 (7, IQR 6-10 vs. 5, IQR 3-6, p < 0.001), CTSS2 (8, IQR 7-11 vs. 5, IQR 4-7, p < 0.001) and POS (2, IQR 1-2.5 vs. 1, IQR 1-2, p < 0.001). Comorbidity (OR = 6.15, p = 0.023) and the presence of low PMI (OR = 5.43, p = 0.001) were predictors of lung injury aggravation with ΔCTSS1 > 4. The presence of low PMI (OR = 5.98, p < 0.001) was the predictor of lung injury aggravation with ΔCTSS2 > 4. Meanwhile, presence of low PMI (OR = 2.82, p = 0.042) and incrementally increasing D-dimer (OR = 0.088, p = 0.024) were predictors of lung injury aggravation with ΔPOS = 2. CONCLUSIONS: PMI can be easily assessed on chest CT images and can potentially be used as one of the markers to predict the severity of lung injury in elderly COVID-19 patients.

A tunable full-color lanthanide noncovalent polymer based on cucurbituril-mediated supramolecular dimerization.

The construction of lanthanide multicolor luminescent materials with tunable photoluminescence properties has been developed as one of the increasingly significant topics and shown inventive applications in miscellaneous fields. However, fabricating such materials based on synergistically assembly-induced emission rather than simple blending of different fluorescent dyes together still remains a challenge. Herein, we report a europium-based noncovalent polymer with tunable full-color emission, which is constructed from the 2,6-pyridinedicarboxylic acid-bearing bromophenylpyridinium salt. This rationally designed bifunctional component can concurrently serve as a guest molecule and a chelating ligand to associate with cucurbit[8]uril and europium ions, thus leading to the formation of a trichromatic (red-green-blue, RGB) photoluminescent polypseudorotaxane-type noncovalent polymer in aqueous solution. Meanwhile, the full-color emission enclosed within the RGB color triangle could be readily produced by simply tuning the molar ratio of cucurbit[8]uril and europium ions. The lanthanide supramolecular polymer featuring tricolor emission, long lifetime, high photoluminescence efficiency and low cytotoxicity could be further applied in multicolor imaging in a cellular environment. These results provide a new and feasible strategy for the construction of full-color single lanthanide self-assembled nanoconstructs.

Effect of 3D Printing Technology in Proximal Femoral Osteotomy in Children with Developmental Dysplasia of the Hip.

OBJECTIVE: To investigate the effect and safety of 3D printing technology in proximal femoral osteotomy in children with developmental dysplasia of the hip. METHODS: 40 cases of children with developmental dysplasia of the hip treated by pelvic osteotomy combined with proximal femoral osteotomy at Ningbo No. 6 Hospital from January 2017 to December 2019 were retrieved and retrospectively analyzed. Among them, 20 cases received preoperative measurement and design assisted by 3D printing technology (the 3D printing group), and 20 cases received conventional preoperative measurement and design (the conventional group). RESULTS: All patients were followed up for an average of 25 (12~36) months. During the follow-up, there were no complications such as infection, fracture of internal fixation, or malunion of osteotomy. Compared with the conventional group, the 3D printing group had a shorter operation time, less intraoperative blood loss, and fewer intraoperative X-ray fluoroscopies (all p < 0.05). In the last follow-up, the clinical efficacy was evaluated by the McKay standard: in the 3D printing group, 14 cases were excellent, 5 cases were good, and 1 case was fair. In the conventional group, 10 cases were excellent, 9 cases were good, and 1 case was fair (Z = -0.382, p > 0.05). CONCLUSION: Preoperative 3D printing of bilateral femur and other large physical models is accurate, which is ideal for the development of individual preoperative planning. Proximal femoral osteotomy using preoperative measurements and simulated surgical data improves the safety of the operation.

[Application of aided three-dimension CT navigation system in craniocervical junction malformation disease surgery].

OBJECTIVE: To investigate the related problems of three-dimension CT navigation system applied to craniocervical junction malformation diseases surgery. METHODS: The clinical data of 61 patients with craniocervical junction malformation disease who underwent surgical treatment from July 2015 to March 2018 was retrospectively analyzed. There were 39 males and 22 females aged from 25 to 73 years old with a mean of (46.20±12.09) years. According to the different ways of screw placements, these patients were divided into navigation group (30 cases) and non navigation group (31 cases). The screws insertion in the navigation group were guided under three-dimension CT computer-aided navigation system, while the screws insertion were guided by C-arm fluoroscopy and intraoperative anatomical landmarks in non-navigation group. All surgeries were performed by the same group of surgeons. All cases were scanned by CT after operation to assess the accuracy of the screw insertion. RESULTS: A total of 122 screws were inserted in the study, including 60 screws in navigation group and 62 screws in non-navigation group. The screw positions in navigation group were all satisfactory, and there were no screw-related complications. The position of 3 screws (4.83%) in non-navigation group was not satisfactory, and there was no obvious neurological symptoms after operation. The operation time of navigation group was from 87.5 to 112.5 min with a mean of (99.6± 12.0) min;and non-navigation group was from 87.5 to 107.5 min, with a mean of (97.5±10.5) min. There was no statistically significant difference in the operation time between two groups (P>0.05). The screw insertion time in navigation group was from 1.50 to 4.20 min, with a mean of (2.51±1.02) min;and non navigation group was from 1.80 to 5.10 min, with a mean of (3.25±1.05) min. The screw insertion time in navigation group was shorter than that in non-navigation group (P<0.05). The intraoperative blood loss in navigation group was from 122 to 153 ml, with a mean of (137.57±9.48) ml, which in non-navigation group wasfrom 121 to 158 ml, with a mean of (138.75±8.56) ml, there was no statistically significant difference between two groups (P>0.05). CONCLUSION: The three-dimensional CT computer-aided navigation system has the characteristics of clear imaging and accurate positioning, which can help improve the accuracy and safety of the screw insertion in craniocervical junction malformation disease surgery.

Supramolecular Assembly with Near-Infrared Emission for Two-Photon Mitochondrial Targeted Imaging.

Two-photon supramolecular assembly with near-infrared (NIR) fluorescence emission is constructed from tetraphenylethene derivative possessing methoxyl and vinyl pyridine salt (TPE-2SP), cucurbit[8]uril (CB[8]), and ß-cyclodextrin modified hyaluronic acid (HA-CD). The obtained experimental results indicate that the TPE-2SP exhibits a very weak fluorescence emission at 650 nm, and then complexes with cucurbit[7]uril (CB[7]) to form 1:2 supramolecular pseudorotaxane with an enhanced NIR fluorescence emission at 660 nm. Compared with CB[7], CB[8] can assemble with TPE-2SP to be two-axial netlike pseudopolyrotaxane, resulting in close packing to increase TPE-2SP fluorescence emission with a redshift of 30 nm. Interestingly, TPE-2SP/CB[8] continues to assemble with cancer cell targeting agent HA-CD into nanoparticles, leading to assembling-induced further enhancement of NIR emission. Surprisingly, supramolecular nanoparticles have the two-photon character, and are successfully applied to mitochondrial targeting imaging. This supramolecular assembly system, with two-photon absorption and assembly-induced enhanced NIR luminescence properties, opens new way for biological targeted imaging.

Type III Monteggia Injury With Ipsilateral Distal Forearm Fracture in a Child: A Case Report.

Monteggia fracture refers to breakage of the upper third of the ulna combined with dislocation of the radial head. It often occurs in children and adolescents and represents a combined injury. Fracture of the distal forearm is among the most common trauma suffered by children. However, distal forearm fractures have rarely been reported as having an association with Monteggia fractures. We report on a 9-year-old boy diagnosed with a type III Monteggia fracture combined with a distal forearm fracture. He underwent surgery and received rehabilitation training 1 month later. He was followed-up for 1 year. The affected limb functioned well with no sign of radial head dislocation.

Bi-needle technique versus transforaminal endoscopic spine system technique for percutaneous endoscopic lumbar discectomy in treating intervertebral disc calcification: a propensity score matched cohort analysis.

OBJECTIVE: The aim of this study was to evaluate the clinical results of a Bi-needle technique and conventional transforaminal endoscopic spine system (TESSYS) technique for percutaneous endoscopic lumbar discectomy (PELD) in treating patients with intervertebral disc calcification (IDC). BACKGROUND: PELD has gained acceptance for treating patients with IDC. The Bi-needle technique was designed to improve the efficiency and safety of PELD. METHOD: Bi-needle and TESSYS group within each cohort were balanced using 1:1 propensity score matching. Finally, 32 patients with IDC treated by Bi-needle technique from December 2015 to September 2017 were enrolled and 25 patients treated by TESSYS technique from the same spine surgery center between January 2013 and October 2017 were enrolled as controls. RESULTS: Propensity score matching generated 22 Bi-needle and 22 TESSYS patients. There were no significant differences in visual analog scale and lumbar Japanese Orthopaedic Association scores between Bi-needle and TESSYS group. Operative time and rate of complications in the Bi-needle was significantly better than the TESSYS group (p < 0.01). CONCLUSIONS: Both surgical methods achieved good clinical outcomes. However, compared with the TESSSY technique, operative time of the Bi-needle technique is shorter, and rate of complications is lower.

[Identification of quality markers for Pudilan Xiaoyan Oral Liquid].

On the basis of anti-inflammatory activity, combined with multiple indicators, the quality markers of Pudilan Xiaoyan Oral Liquid were screened and determined for quality control. Lipopolysaccharide(LPS) was used to induce normal human bronchial epithelial cell(NHBE) inflammation model. The anti-inflammatory effects of the main chemical components in Pudilan Xiaoyan Oral Liquid were examined one by one, and the pharmacodynamic basis for the overall anti-inflammatory efficacy of Pudilan Xiaoyan Oral Liquid was clarified to identify the quality markers of Pudilan Xiaoyan Oral Liquid and the contents of the quality markers were determined by high performance liquid chromatography(HPLC). The results showed that adenosine, epigoitrin, chlorogenic acid, caffeic acid, cichoric acid, corynoline, baicalin, wogonoside, wogonin and oroxylin A had a certain regulatory effect on inflammatory factor tumor necrosis factor(TNF-α), interleukin(IL-1ß) and IL-6 at specific concentrations in a dose-dependent manner. Considering the factors such as the IC_(50) value of each monomer component and the comprehensiveness of the quality control components, we proposed to use adenosine, cichoric acid, corynoline, baicalin and wogonin as quality markers of Pudilan Xiaoyan Oral Liquid. The contents of the five components were determined by HPLC, and the results showed that they were relatively stable in three batches of Pudilan Xiaoyan Oral Liquid. In this study, the quality control components selected by the anti-inflammatory activity test have a clear material basis, covering all four active pharmaceutical ingredients, which can fully reflect the quality of Pudilan Xiaoyan Oral Liquid, and effectively improve the quality control standard of Pudilan Xiaoyan Oral Liquid.

LEF1-AS1 is implicated in the malignant development of glioblastoma via sponging miR-543 to upregulate EN2.

Glioblastoma (GBM) has been regarded as the most aggressive disease in the nervous system. Accumulating literatures have illustrated the crucial role of competing endogenous RNAs (ceRNAs) network in the pathogenesis and progression of various tumors. The promoting effect of LEF1-AS1 on GBM development has been previously identified. This study attempted to explore the underlying mechanism of LEF1-AS1 in GBM. Data of clinical GBM patients was downloaded from TCGA and GEO databases. The proliferative ability, clonogenic vitality, invasive, and migratory capabilities of GBM cells were measured using Cell counting kit-8 (CCK-8), colony formation and transwell assays. Luciferase reporter gene analysis was performed to verify the correlations between LEF1-AS1/EN2 and miR-543. qRT-PCR and western blotting were implemented to evaluate the mRNA and protein levels, respectively. Our results consolidated that LEF1-AS1 was highly expressed in GBM tissue specimens and its up-regulation induced unfavorable prognosis. The loss/gain-of-function analyses verified that LEF1-AS1 promoted the GBM cell malignant behaviors. Mechanically, LEF1-AS1 acted as a ceRNA for miR-543 and positively regulated engrailed homeobox 2 (EN2) expression. Down-regulation of miR-543 elevated GBM cell malignant behaviors, which was reversed by LEF1-AS1 knockdown. Meanwhile, the LEF1-AS1 inhibition could arrest the promoting effect of high-regulated EN2 on GBM cell aggressiveness and vice versa. In conclusion, our findings identified LEF1-AS1 as a ceRNA for miR-543 and showed that EN2 was positively regulated by LEF1-AS1. The LEF1-AS1/miR-543/EN2, as a novel ceRNA network, was implicated in the progression of GBM, which provided a novel insight for GBM treatment.

Highly efficient photocontrolled targeted delivery of siRNA by a cyclodextrin-based supramolecular nanoassembly.

In this study, we have constructed a binary supramolecular nanoassembly composed of α-cyclodextrin-modified hyaluronic acid and an azobenzene-modified diphenylalanine derivative with a positively charged imidazole group. This nanoassembly can bind with siRNA through electrostatic interactions and efficiently delivered them into cancer cells and inhibited their growth.

Twisting Steel Wires with a Drill: A New Surgical Technique.

Stainless steel wires are often used to fix specific types of fractures in orthopaedic surgery using pliers. This article aims to introduce a new technique to twist steel wires by using an electric drill. The steps before twisting the wire are the same as usual. Our technique is as follows. First, tighten the two ends of wire by using pliers, then insert both the ends of wires into the drill and hold in place. Second, set the drill to reverse mode. Third, start turning the drill, then the two wires begin to intertwine and tighten. It is important to stop turning before the wire (the twisted part) begins to bend. Finally, cut the twisted part of wire in place, and bend the wire stump. This technique can achieve a better appearance while saving the strength of surgeons. It has the same clinical effect as the traditional method. This technique provides a new method for surgery and wide clinical application.

Mechanism of interaction between ocular and nasal neurogenic inflammation in allergic rhinoconjunctivitis.

PURPOSE: The mechanisms of naso-ocular interaction in allergic rhinoconjunctivitis are not well understood. Neurogenic inflammation affects both eyes and nose via the same neurogenic factors. The purpose of this study was to investigate the effects of neurogenic inflammation on conjunctival inflammation following nasal allergen provocation. METHODS: Sensitized rats were exposed to ovalbumin (OVA) via the nose. Parts of the nasal mucosa and conjunctivae were sliced and used for hematoxylin-eosin staining, immunohistochemical analysis, western blotting, and real-time polymerase chain reaction. The slides were observed under a light microscope, and the acquired images were analyzed. The levels of substance P (SP), vasoactive intestinal peptide (VIP), and nerve growth factor (NGF) were detected. RESULTS: The levels of SP, VIP, and NGF were increased in both nasal mucosa and conjunctivae 1 h and 24 h after OVA administration (p < 0.05). Higher levels of SP, VIP, and NGF expression were observed in the nasal mucosa and conjunctivae 24 h after OVA administration (p < 0.05). Following damage of the nasal sensory nerves by capsaicin, the protein and mRNA levels of SP, VIP, and NGF were reduced. CONCLUSION: In conclusion, the increased levels of VIP, SP, and NGF might be responsible for the ocular reaction following nasal challenge with allergen in rats.