Patient-derived melanoma organoid models facilitate the assessment of immunotherapies.

BACKGROUND: Only a minority of melanoma patients experience durable responses to immunotherapies due to inter- and intra-tumoral heterogeneity in melanoma. As a result, there is a pressing need for suitable preclinical models to investigate resistance mechanisms and enhance treatment efficacy. METHODS: Here, we report two different methods for generating melanoma patient-derived organoids (MPDOs), one is embedded in collagen gel, and the other is inlaid in Matrigel. MPDOs in Matrigel are used for assessing the therapeutic effects of anti-PD-1 antibodies (αPD-1), autochthonous tumor infiltrating lymphocytes (TILs), and small molecule compounds. MPDOs in collagen gel are used for evaluating the chemotaxis and migratory capacity of TILs. FINDING: The MPDOs in collagen gel and Matrigel have similar morphology and immune cell composition to their parental melanoma tissues. MPDOs show inter- and intra-tumoral heterogeneity and contain diverse immune cells such as CD4+, CD8+ T, Treg, CD14+ monocytic, CD15+, and CD11b+ myeloid cells. The tumor microenvironment (TME) in MPDOs is highly immunosuppressive, and the lymphoid and myeloid lineages express similar levels of PD-1, PD-L1, and CTLA-4 as their parental melanoma tissues. Anti-PD-1 antibodies (αPD-1) reinvigorate CD8+ T cells and induce melanoma cell death in the MPDOs. TILs expanded by IL-2 and αPD-1 show significantly lower expression of TIM-3, better migratory capacity and infiltration of autochthonous MPDOs, and more effective killing of melanoma cells than TILs expanded by IL-2 alone or IL-2 with αCD3. A small molecule screen discovers that Navitoclax increases the cytotoxicity of TIL therapy. INTERPRETATION: MPDOs may be used to test immune checkpoint inhibitors and cellular and targeted therapies. FUNDING: This work was supported by the NIH grants CA114046, CA261608, CA258113, and the Tara Miller Melanoma Foundation.

Efficacy and safety of Tuina for chronic nonspecific low back pain: A PRISMA-compliant systematic review and meta-analysis.

OBJECTIVE: Chronic nonspecific low back pain (CNLBP) is a serious medical and social problem resulting in functional decline and decreased work ability. Tuina, a form of manual therapy, has been sparsely used to treat patients with CNLBP. To systematically assess the efficacy and safety of Tuina for patients with CNLBP. METHODS: Multiple English and Chinese literature databases were searched until September 2022 for randomized controlled trials (RCTs) of Tuina in the treatment of CNLBP. The methodological quality was assessed using the Cochrane Collaboration's tool, and certainty of the evidence was determined with the online Grading of Recommendations, Assessment, Development and Evaluation tool. RESULTS: Fifteen RCTs with 1390 patients were included. Tuina demonstrated a significant effect on pain (SMD: -0.82; 95% CI -1.12 to -0.53; P < .001; I2 = 81%) and physical function (SMD: -0.91; 95% CI -1.55 to -0.27; P = .005; I2 = 90%) when compared to control. However, Tuina resulted in no significant improvement for quality of life (QoL) (SMD: 0.58; 95% CI -0.04 to 1.21; P = .07; I2 = 73%;) compared to control. The Grading of Recommendations, Assessment, Development and Evaluation evidence quality was determined to be low level for pain relief, physical function, and QoL measurements. Only six studies reported adverse events; none were serious. CONCLUSION: Tuina might be an effective and safe strategy for treating CNLBP in terms of pain and physical function, but not for QoL. The study results should be interpreted with caution for their low-level evidence. More multicenter, large-scale RCTs with a rigorous design are required to further confirm our findings.

Effect of high-frequency repetitive transcranial magnetic stimulation over M1 for consciousness recovery after traumatic brain injury.

BACKGROUND: The brain area stimulated during repetitive transcranial magnetic stimulation (rTMS) treatment is important in altered states of consciousness. However, the functional contribution of the M1 region during the treatment of high-frequency rTMS remains unclear. OBJECTIVE: The aim of this study was to examine the clinical [the Glasgow coma scale (GCS) and the coma recovery scale-revised (CRS-R)] and neurophysiological (EEG reactivity and SSEP) responses in vegetative state (VS) patients following traumatic brain injury (TBI) before and after a protocol of high-frequency rTMS over the M1 region. METHODS: Ninety-nine patients in a VS following TBI were recruited so that their clinical and neurophysiological responses could be evaluated in this study. These patients were randomly allocated into three experimental groups: rTMS over the M1 region (test group; n = 33), rTMS over the left dorsolateral prefrontal cortex (DLPFC) (control group; n = 33) and placebo rTMS over the M1 region (placebo group; n = 33). Each rTMS treatment lasted 20 min and was carried out once a day. The duration of this protocol was a month with 20 treatments (5 times per week) occurring with that time. RESULTS: We found that the clinical and neurophysiological responses improved after treatment in the test, control, and placebo groups; the improvement was highest in the test group compared to that in the control and placebo groups. CONCLUSIONS: Our results demonstrate an effective method of high-frequency rTMS over the M1 region for consciousness recovery after severe brain injury.

Plantar pressure and falling risk in older individuals: a cross-sectional study.

BACKGROUND: Falls are commonplace among elderly people. It is urgent to prevent falls. Previous studies have confirmed that there is a difference in plantar pressure between falls and non-falls in elderly people, but the relationship between fall risk and foot pressure has not been studied. In this study, the differences in dynamic plantar pressure between elderly people with high and low fall risk were preliminarily discussed, and the characteristic parameters of plantar pressure were determined. METHODS: Twenty four high-fall-risk elderly individuals (HR) and 24 low-fall-risk elderly individuals (LR) were selected using the Berg Balance Scale 40 score. They wore wearable foot pressure devices to walk along a 20-m-long corridor. The peak pressure (PP), pressure time integral (PTI), pressure gradient (maximum pressure gradient (MaxPG), minimum pressure gradient (MinPG), full width at half maximum (FWHM)) and average pressure (AP) of their feet were measured for inter-group and intra-group analysis. RESULTS: The foot pressure difference comparing the high fall risk with low fall risk groups was manifested in PP and MaxPG, concentrated in the midfoot and heel (p < 0.05), while the only time parameter, FWHM, was manifested in the whole foot (p < 0.05). The differences between the left and right foot were reflected in all parameters. The differences between the left and right foot in LR were mainly reflected in the heel (p < 0.05), while it in the HR was mainly reflected in the forefoot (p < 0.05). CONCLUSIONS: The differences comparing the high fall risk with low fall risk groups were mostly reflected in the midfoot and heel. The HR may have been more cautious when landing. In the intra-group comparison, the difference between the right and left foot of the LR was mainly reflected during heel striking, while it was mainly reflected during pedalling in the HR. The sensitivity of PP, PTI and AP was lower and the newly introduced pressure gradient could better reflect the difference in foot pressure between the two groups. The pressure gradient can be used as a new foot pressure parameter in scientific research.

Novel Approach to Transfer of Coordination Skills to the Paretic Hand by Improving the Efficiency of Cortical Network.

We have previously shown that healthy subjects can transfer coordination skills to the unpracticed hand by performing a unimanual task with the other hand and visualizing a bimanual action using a game-like interactive system. However, whether this system could be used to transfer coordination skills to the paretic hand after stroke and its underlying neural mechanism remain unknown. Here, using a game-like interactive system for visualization during physical practice in an immersive virtual reality environment, we examined coordination skill improvement in the unpracticed/paretic hand after training in 10 healthy subjects and 13 chronic and sub-acute stroke patients. The bimanual movement task was defined as simultaneously drawing non-symmetric three-sided squares (e.g., U and C), while the training strategy was performing a unimanual task with the right/nonparetic hand and visualizing a bimanual action. We found large decreases in the intra-hand temporal and spatial measures for movement in the unpracticed/paretic hand after training. Furthermore, a substantial reduction in the inter-hand temporal and spatial interference was observed after training. Additionally, we examined the related cortical network evolution using EEG in both the healthy subjects and stroke patients. Our studies show that the cortical network became more efficient after training in the healthy subjects and stroke patients. These results demonstrate that our proposed method could contribute to the transference of coordination skill to the paretic/unpracticed hand by promoting the efficiency of cortical networks.

A Systematic Review and Meta-Analysis Protocol on How Best to Use Non-Pharmacologic Therapies to Manage Chronic Low Back Pain and Associated Depression.

Purpose: To evaluate the efficacy of different non-pharmacologic therapies (NPTs) on relieving depressive symptoms and pain intensity in individuals living with chronic low back pain (LBP) and associated depression. Methods: A comprehensive search of seven English databases and two Chinese databases from inception to the search date will be undertaken. The reference lists of previously published relevant reviews and included trials will also be searched. Only peer-reviewed and published moderate-to-high quality randomized controlled trials (RCTs) for chronic LBP and associated depression treated with NPTs will be considered. Two independent reviewers will identify studies, extract data, assess risk of bias, and evaluate the strength of evidence using the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) approach. Meta-analysis will be conducted to estimate the treatment effect of various NPTs. Heterogeneity will be assessed using Cochrane's Q and the I-squared statistics. Subgroup and sensitivity analyses will be performed to assess the robustness of findings. A funnel plot will be developed to evaluate reporting bias, and Begg's and Egger's tests will be used to assess funnel plot symmetries. Results: This protocol outlines the planned scope and methodology for an upcoming systematic review and meta-analysis, which will provide up-to-date evidence on 1) which NPTs are associated with improvements in depressive symptoms and pain intensity and 2) whether the effects of NPTs on chronic LBP and associated depression vary according to clinical condition, participant, and treatment characteristics. Conclusion: Our meta-analyses of moderate-to-high quality RCTs will help to develop specific recommendations on prescribing NPTs in patients with chronic LBP and associated depression. Study Registration: This protocol is registered on the International Platform of Registered Systematic Review and Meta-analysis (INPLASY) protocols platform as record No. INPLASY202260055.

The Use of Antibiotics for Ventilator-Associated Pneumonia in the MIMIC-IV Database.

Purpose: By analyzing the clinical characteristics, etiological characteristics and commonly used antibiotics of patients with ventilator-associated pneumonia (VAP) in intensive care units (ICUs) in the intensive care database. This study aims to provide guidance information for the clinical rational use of drugs for patients with VAP. Method: Patients with VAP information were collected from the Medical Information Mart for Intensive Care IV (MIMIC-IV) database, including their sociodemographic characteristics, vital signs, laboratory measurements, complications, microbiology, and antibiotic use. After data processing, the characteristics of the medications used by patients with VAP in ICUs were described using statistical graphs and tables, and experiences were summarized and the reasons were analyzed. Results: This study included 2,068 patients with VAP. Forty-eight patient characteristics, including demographic indicators, vital signs, biochemical indicators, scores, and comorbidities, were compared between the survival and death groups of VAP patients. Cephalosporins and vancomycin were the most commonly used. Among them, fourth-generation cephalosporin (ForGC) combined with vancomycin was used the most, by 540 patients. First-generati49n cephalosporin (FirGC) combined with vancomycin was associated with the highest survival rate (86.7%). More than 55% of patients were infected with Gram-negative bacteria. However, patients with VAP had fewer resistant strains (<25%). FirGC or ForGC combined with vancomycin had many inflammation-related features that differed significantly from those in patients who did not receive medication. Conclusion: Understanding antibiotic use, pathogenic bacteria compositions, and the drug resistance rates of patients with VAP can help prevent the occurrence of diseases, contain infections as soon as possible, and promote the recovery of patients.

Deep transfer learning to quantify pleural effusion severity in chest X-rays.

PURPOSE: The detection of pleural effusion in chest radiography is crucial for doctors to make timely treatment decisions for patients with chronic obstructive pulmonary disease. We used the MIMIC-CXR database to develop a deep learning model to quantify pleural effusion severity in chest radiographs. METHODS: The Medical Information Mart for Intensive Care Chest X-ray (MIMIC-CXR) dataset was divided into patients 'with' or 'without' chronic obstructive pulmonary disease (COPD). The label of pleural effusion severity was obtained from the extracted COPD radiology reports and classified into four categories: no effusion, small effusion, moderate effusion, and large effusion. A total of 200 datasets were randomly sampled to manually check each item and determine whether the tags are correct. A professional doctor re-tagged these items as a verification cohort without knowing their previous tags. The learning models include eight common network structures including Resnet, DenseNet, and GoogleNET. Three data processing methods (no sampling, downsampling, and upsampling) and two loss algorithms (focal loss and cross-entropy loss) were used for unbalanced data. The Neural Network Intelligence tool was applied to train the model. Receiver operating characteristic curves, Area under the curve, and confusion matrix were employed to evaluate the model results. Grad-CAM was used for model interpretation. RESULTS: Among the 8533 patients, 15,620 chest X-rays with clearly marked pleural effusion severity were obtained (no effusion, 5685; small effusion, 4877; moderate effusion, 3657; and large effusion, 1401). The error rate of the manual check label was 6.5%, and the error rate of the doctor's relabeling was 11.0%. The highest accuracy rate of the optimized model was 73.07. The micro-average AUCs of the testing and validation cohorts was 0.89 and 0.90, respectively, and their macro-average AUCs were 0.86 and 0.89, respectively. The AUC of the distinguishing results of each class and the other three classes were 0.95 and 0.94, 0.76 and 0.83, 0.85 and 0.83, and 0.87 and 0.93. CONCLUSION: The deep transfer learning model can grade the severity of pleural effusion.

The effectiveness of Tuina in managing chronic non-specific low back pain: A protocol of a multicenter international randomized controlled trial.

BACKGROUND: Chronic non-specific low back pain (CNLBP) is a common complaint about medical care and carries a heavy social burden. The efficacy of Tuina (TN) or physiotherapy (PT) for CNLBP has been evaluated in previous systematic reviews. However, there is no high-quality evidence to support the efficacy of Tuina. Therefore, this study aims to conduct a large-scale, multicenter, high-quality clinical trial to provide evidence for Tuina to treat CNLBP. METHODS: This is a multicenter, assessor-, and analyst-blinded, randomized controlled trial with 3 parallel arms: TN, PT, and TN combined with PT (Tuina combined with physiotherapy) group. Six hundred twelve eligible CNLBP patients will be randomly assigned to the groups in a 1:1:1 ratio in 3 centers. The TN intervention includes 9-step routine techniques, while the PT intervention includes a physiotherapy treatment plan based on a patient's symptoms. The interventions for both groups will last for 30 minutes and will be carried out for 6 sessions in 8 weeks. The primary outcome will be the visual analog scale pain score. And the secondary outcomes will include the Oswestry Disability Index, spinal range of motion, 36-item short-form health survey. Safety evaluation will be recorded during the whole study. All data in this randomized controlled trial will be analyzed by SAS 9.4. DISCUSSION: The results of this trial will provide evidence to evaluate the efficacy of Tuina's value as a treatment for CNLBP. TRIAL REGISTRATION: Chinese Clinical Trial Registry (ChiCTR2000040288, November 27, 2020).

A novel upper-limb tracking system in a virtual environment for stroke rehabilitation.

BACKGROUND: The transfer of the behaviors of a human's upper limbs to an avatar is widely used in the field of virtual reality rehabilitation. To perform the transfer, movement tracking technology is required. Traditionally, wearable tracking devices are used for tracking; however, these devices are expensive and cumbersome. Recently, non-wearable upper-limb tracking solutions have been proposed, which are less expensive and more comfortable. However, most products cannot track the upper limbs, including the arms and all the fingers at the same time, which limits the limb parts for tracking in a virtual environment and may lead to a limited rehabilitation effect. METHODS: In this paper, a novel virtual reality rehabilitation system (VRRS) was developed for upper-limb rehabilitation. The VRRS could track the motion of both upper limbs, integrate fine finger motion and the range of motion of the entire arm and map the motion to an avatar. To test the performance of VRRS, two experiments were designed. In the first experiment, we investigated the effect of VRRS on virtual body ownership, agency and location of the body and usability in 8 healthy participants by comparing it with a partial upper-limb tracking method based on a Leap Motion controller (LP) in the same virtual environments. In the second experiment, we examined the feasibility of VRRS in upper-limb rehabilitation with 27 stroke patients. RESULTS: VRRS improved the users' senses of body ownership, agency, and location of the body. The users preferred using the VRRS to using the LP. In addition, we found that although the upper limb motor function of patients from all groups was improved, the difference between the FM scores tested on the first day and the last day of the experimental group was more significant than that of the control groups. CONCLUSIONS: A VRRS with motion tracking of the upper limbs and avatar control including the arms and all the fingers was developed. It resulted in an improved user experience of embodiment and effectively improved the effects of upper limb rehabilitation in stroke patients. TRIAL REGISTRATION: The study was registered at the First Affiliated Hospital of Jinan University Identifier: KY-2020-036; Date of registration: June 01, 2020.

Tui Na for Chronic Nonspecific Low Back Pain: Protocol for a Systematic Review and Meta-analysis.

BACKGROUND: Chronic nonspecific low back pain (CNLBP) is one of the most common complex pain conditions, and it is strongly associated with high rates of disability. Even though several studies on Tui na for CNLBP have been reported, to our knowledge there has been no systematic review of the currently available publications. OBJECTIVE: This study aims to develop a protocol for a systematic review and meta-analysis that will evaluate the effectiveness and safety of Tui na therapy for patients with CNLBP. METHODS: An electronic literature search of PubMed, Embase, MEDLINE, Cochrane Library, Springer, Scopus, World Health Organization International Clinical Trials Registry Platform, Physiotherapy Evidence Database (PEDro), Clarivate Analytics, and Chinese biomedical databases (the China National Knowledge Infrastructure, Wan-fang database, Chinese Scientific Journals Database, and Chinese Biomedical Literature Databases) will be conducted. Studies will be screened by two reviewers independently based on titles and abstracts, followed by a full-text reading with eligibility criteria. Randomized controlled trials involving Tui na for patients with CNLBP will be reviewed. The primary outcomes of the study are improvement of pain, analgesic medication reduction, improvement of functional disability, and degree of satisfaction with the intervention. A secondary outcome is any adverse event of Tui na intervention. Methodological quality and risk of bias will be assessed with the Cochrane Collaboration Risk of Bias Tool. If studies are sufficient, a meta-analysis of the effectiveness will be performed. If possible, we will evaluate publication bias using funnel plots. If substantial heterogeneity between studies is present, and there are sufficient studies, subgroup analyses will be conducted to explain the study findings. RESULTS: The review database searches will be initiated in December 2020, with findings expected by January 2021. CONCLUSIONS: This protocol will establish a framework of a high-quality literature synthesis on the impact of Tui na treatment in patients with CNLBP. The proposed review will determine whether Tui na is effective and safe for CNLBP patients. TRIAL REGISTRATION: PROSPERO CRD42020166731; https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=166731. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): PRR1-10.2196/20615.

Genome-Wide Analysis, Expression Profile, and Characterization of the Acid Invertase Gene Family in Pepper.

Catalytic decomposition of sucrose by acid invertases (AINVs) under acidic conditions plays an important role in the development of sink organs in plants. To reveal the function of AINVs in the development of pepper fruits, nine AINV genes of pepper were identified. Protein sequencing and phylogenetic analysis revealed that the CaAINV family may be divided into cell wall invertases (CaCWINV1⁻7) and vacuolar invertases (CaVINV1⁻2). CaAINVs contain conserved regions and protein structures typical of the AINVs in other plants. Gene expression profiling indicated that CaCWINV2 and CaVINV1 were highly expressed in reproductive organs but differed in expression pattern. CaCWINV2 was mainly expressed in buds and flowers, while CaVINV1 was expressed in developmental stages, such as the post-breaker stage. Furthermore, invertase activity of CaCWINV2 and CaVINV1 was identified via functional complementation in an invertase-deficient yeast. Optimum pH for CaCWINV2 and CaVINV1 was found to be 4.0 and 4.5, respectively. Gene expression and enzymatic activity of CaCWINV2 and CaVINV1 indicate that these AINV enzymes may be pivotal for sucrose hydrolysis in the reproductive organs of pepper.

Genome-Wide Identification, Expression, and Functional Analysis of the Alkaline/Neutral Invertase Gene Family in Pepper.

Alkaline/neutral invertase (NINV) proteins irreversibly cleave sucrose into fructose and glucose, and play important roles in carbohydrate metabolism and plant development. To investigate the role of NINVs in the development of pepper fruits, seven NINV genes (CaNINV1-7) were identified. Phylogenetic analysis revealed that the CaNINV family could be divided into α and ß groups. CaNINV1-6 had typical conserved regions and similar protein structures to the NINVs of other plants, while CaNINV7 lacked amino acid sequences at the C-terminus and N-terminus ends. An expression analysis of the CaNINV genes in different tissues demonstrated that CaNINV5 is the dominant NINV in all the examined tissues (root, stem, leaf, bud, flower, and developmental pepper fruits stage). Notably, the expression of CaNINV5 was found to gradually increase at the pre-breaker stages, followed by a decrease at the breaker stages, while it maintained a low level at the post-breaker stages. Furthermore, the invertase activity of CaNINV5 was identified by functional complementation of the invertase-deficient yeast strain SEY2102, and the optimum pH of CaNINV5 was found to be ~7.5. The gene expression and enzymatic activity of CaNINV5 suggest that it might be the main NINV enzyme for hydrolysis of sucrose during pepper fruit development.

SpBADH of the halophyte Sesuvium portulacastrum strongly confers drought tolerance through ROS scavenging in transgenic Arabidopsis.

Glycine betaine (GB) accumulation is involved in abiotic stress. However, it is not known whether BADH, the key enzyme of GB synthesis, utilizes the antioxidant system to confer drought stress tolerance. In this study, a novel member of the ALDH10 gene family, SpBADH, was isolated from Sesuvium portulacastrum. The expression of this gene was up-regulated by NaCl, PEG6000, H2O2, ABA and high temperature in S. portulacastrum. SpBADH overexpression in Arabidopsis resulted in higher BADH activity and GB content and might increase tolerance to drought/osmotic stresses, specifically strong tolerance to drought stress. Transgenic lines exhibited lower MDA and H2O2 contents but higher proline, POD, SOD and CAT contents than the wild type under drought and osmotic stresses. SpBADH overexpression in Arabidopsis also enhanced the expression of ROS-related genes including AtSOD, AtPOD, AtCAT, AtAPX and Atpsb under drought and osmotic stresses. Thus, SpBADH increases plant tolerance to drought or osmotic stresses by reducing H2O2, increasing proline, and activating antioxidative enzymes to improve ROS scavenging.