Novel Near-Infrared Fluorescent Probe for Hepatocyte Growth Factor in Vivo Imaging in Surgical Navigation of Colorectal Cancer.

Despite recent advancements in colorectal cancer (CRC) treatment, the prognosis remains unfavorable primarily due to high recurrence and liver metastasis rates. Fluorescence molecular imaging technologies, combined with specific probes, have gained prominence in facilitating real-time tumor resection guided by fluorescence. Hepatocyte growth factor (HGF) is overexpressed in CRC, but the advancement of HGF fluorescent probes has been impeded by the absence of effective HGF-targeting small-molecular ligands. Herein, we present the targeted capabilities of the novel V-1-GGGK-MPA probe labeled with a near-infrared fluorescent dye, which targets HGF in CRC. The V-1-GGGK peptide exhibits high specificity and selectivity for HGF-positive in vitro tumor cells and in vivo tumors. Biodistribution analysis of V-1-GGGK-MPA revealed tumor-specific accumulation with low background uptake, yielding signal-to-noise ratio (SNR) values of tumor-to-colorectal >6 in multiple subcutaneous CRC models 12 h postinjection. Quantitative analysis confirmed the probe's high uptake in SW480 and HT29 orthotopic and liver metastatic models, with SNR values of tumor-to-colorectal and -liver being 5.6 ± 0.4, 4.6 ± 0.5, and 2.1 ± 0.3, 2.0 ± 0.5, respectively, enabling precise tumor visualization for surgical navigation. Pathological analysis demonstrated the excellent tumor boundaries discrimination capacity of the V-1-GGGK-MPA probe at the molecular level. With its rapid tumor targeting, sustained tumor retention, and precise tumor boundary delineation, V-1-GGGK-MPA merges as a promising HGF imaging agent, enriching the toolbox of intraoperative navigational fluorescent probes for CRC.

Effects of proprioceptive neuromuscular facilitation combined with threshold inspiratory muscle training on respiratory function in neurocritical patients with weaning failure: a randomized controlled trial.

The purpose of this study was to determine the effects of combining proprioceptive neuromuscular facilitation (PNF) with threshold inspiratory muscle training (TIMT), compared with TIMT alone, on respiratory function in neurocritical patients who experienced a weaning failure. Forty-seven participants (mostly after a stroke), were randomly divided into the experimental group (n = 24) and the control group (n = 23). The control group received usual care and TIMT, whereas the experimental group, in addition, underwent four 90-s periods of manual PNF. Both groups performed training in the ICU twice a day for 5 consecutive days. The main outcome measures included maximum inspiratory pressure, diaphragmatic excursions, diaphragm thickening fraction, oxygenation index, and forced expiratory volume in 1 s/forced vital capacity. The results showed a significant group-by-time interaction effect for maximum inspiratory pressure [F (1, 45) = 17.84, η2 = 0.328, P < 0.001] and oxygenation index [F [1, 45) = 5.58, η2 = 0.11, P = 0.023]. When compared with the control group, the experimental group showed overall significantly higher maximum inspiratory pressure [mean difference = 4.37 cm H2O, 95% confidence interval (CI) 0.25-8.50, P = 0.038]. No other significant group differences were found. Combining PNF with TIMT may improve respiratory function in neurocritical patients with weaning failure. This combination approach may increase the likelihood of survival of neurocritical patients in the ICU.

Development and validation of a nomogram model for predicting unfavorable functional outcomes in ischemic stroke patients after acute phase.

Introduction: Prediction of post-stroke functional outcome is important for personalized rehabilitation treatment, we aimed to develop an effective nomogram for predicting long-term unfavorable functional outcomes in ischemic stroke patients after acute phase. Methods: We retrospectively analyzed clinical data, rehabilitation data, and longitudinal follow-up data from ischemic stroke patients who underwent early rehabilitation at multiple centers in China. An unfavorable functional outcome was defined as a modified Rankin Scale (mRS) score of 3-6 at 90 days after onset. Patients were randomly allocated to either a training or test cohort in a ratio of 4:1. Univariate and multivariate logistic regression analyses were used to identify the predictors for the development of a predictive nomogram. The area under the receiver operating characteristic curve (AUC) was used to evaluate predictive ability in both the training and test cohorts. Results: A total of 856 patients (training cohort: n = 684; test cohort: n = 172) were included in this study. Among them, 518 patients experienced unfavorable outcomes 90 days after ischemic stroke. Trial of ORG 10172 in Acute Stroke Treatment classification (p = 0.024), antihypertensive agents use [odds ratio (OR) = 1.86; p = 0.041], 15-day Barthel Index score (OR = 0.930; p < 0.001) and 15-day mRS score (OR = 13.494; p < 0.001) were selected as predictors for the unfavorable outcome nomogram. The nomogram model showed good predictive performance in both the training (AUC = 0.950) and test cohorts (AUC = 0.942). Conclusion: The constructed nomogram model could be a practical tool for predicting unfavorable functional outcomes in ischemic stroke patients underwent early rehabilitation after acute phase.

Analysis of Resting Energy Consumption and Its Influencing Factors in Stroke Patients with Severe Neurological Diseases: A Retrospective Clinical Study.

BACKGROUND: This study aimed to observe the changes of resting energy metabolism in patients with severe neurological diseases, and to explore the effects of tracheostomy status, stroke severity, and complications on resting energy expenditure (REE) and respiratory quotient (RQ). METHODS: A retrospective study was conducted in 105 patients with neurological rehabilitation who were hospitalized in the Rehabilitation Department of the Affiliated Jiangning Hospital of Nanjing Medical University from August 2018 to October 2021. REE was measured by Italian Cosmed k4b2 indirectly, and white blood cell count and C-reactive protein (CRP) were collected. RESULTS: Among the 105 patients, there were 18 cases of mild stroke, 45 cases of moderate stroke, and 42 cases of severe stroke. The difference between predicted REE and actual REE among different degrees of stroke patients was statistically significant (p < 0.05); there was no significant difference in RQ values among different degrees of stroke patients (p > 0.05). Hemoglobin, albumin, and body mass index were significantly and positively correlated with predicted REE and actual REE, while CRP was significantly negatively correlated with predicted REE and actual REE. There was no significant difference in predicted REE, actual REE, and RQ between renal insufficiency, type 2 diabetes mellitus, and chronic obstructive pulmonary disease (p > 0.05). The CRP level could affect the REE of stroke patients. CONCLUSION: Metabolic vehicle assay has a certain clinical value in accurately evaluating the metabolic needs and feeding level of patients.

ALICE: a hybrid AI paradigm with enhanced connectivity and cybersecurity for a serendipitous encounter with circulating hybrid cells.

A fully automated and accurate assay of rare cell phenotypes in densely-packed fluorescently-labeled liquid biopsy images remains elusive. Methods: Employing a hybrid artificial intelligence (AI) paradigm that combines traditional rule-based morphological manipulations with modern statistical machine learning, we deployed a next generation software, ALICE (Automated Liquid Biopsy Cell Enumerator) to identify and enumerate minute amounts of tumor cell phenotypes bestrewed in massive populations of leukocytes. As a code designed for futurity, ALICE is armed with internet of things (IOT) connectivity to promote pedagogy and continuing education and also, an advanced cybersecurity system to safeguard against digital attacks from malicious data tampering. Results: By combining robust principal component analysis, random forest classifier and cubic support vector machine, ALICE was able to detect synthetic, anomalous and tampered input images with an average recall and precision of 0.840 and 0.752, respectively. In terms of phenotyping enumeration, ALICE was able to enumerate various circulating tumor cell (CTC) phenotypes with a reliability ranging from 0.725 (substantial agreement) to 0.961 (almost perfect) as compared to human analysts. Further, two subpopulations of circulating hybrid cells (CHCs) were serendipitously discovered and labeled as CHC-1 (DAPI+/CD45+/E-cadherin+/vimentin-) and CHC-2 (DAPI+ /CD45+/E-cadherin+/vimentin+) in the peripheral blood of pancreatic cancer patients. CHC-1 was found to correlate with nodal staging and was able to classify lymph node metastasis with a sensitivity of 0.615 (95% CI: 0.374-0.898) and specificity of 1.000 (95% CI: 1.000-1.000). Conclusion: This study presented a machine-learning-augmented rule-based hybrid AI algorithm with enhanced cybersecurity and connectivity for the automatic and flexibly-adapting enumeration of cellular liquid biopsies. ALICE has the potential to be used in a clinical setting for an accurate and reliable enumeration of CTC phenotypes.

Fabrication and characterization of high molecular keratin based nanofibrous membranes for wound healing.

Keratin is widely used in the biomaterial application, but the keratin prepared by the physical or chemical approach has relatively low molecular weight and mechanical properties. Here we report the preparation of high molecular keratin (HMK) with molecular weight of 120 kDa via multi-enzyme cascade pathway and its application in wound healing. Briefly, we prepared the soluble keratin from wool by keratinase and improved the molecular weight of keratin by transglutaminase (TGase). The HMK was coelectrospun with poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) and the prepared nanofibrous mats demonstrated improved mechanical properties. Ag nanoparticles (AgNPs) were synthesized on the nanofibers via in situ bioreduction, using the above-mentioned keratinase as the reducing agent. It is demonstrated that the PHBV/HMK/AgNPs nanofibrous mats possess favorable antibacterial properties and good biocompatibility. Moreover, in vivo wound healing assessment, the PHBV/HMK/AgNPs membrane displayed better wound healing ability than the control group. These results indicate that PHBV/HMK/AgNPs mats exhibit significant potential in tissue engineering.

Osw2 is required for proper assembly of glucan and/or mannan layers of the yeast spore wall.

OSW2 is a meiotically-induced gene required for spore wall formation. osw2Δ spores are sensitive to ether treatment. Except for this phenotype, the mutants do not show obvious sporulation defects; thus, its function remains elusive. We found that deletion of both OSW2 and CHS3 results in a synthetic sporulation defect. The spore wall is composed of four layers, and chs3Δ spores lack the outer two (chitosan and dityrosine) layers. Thus, Osw2 is involved in the assembly of the inner (glucan and mannan) layers. In agreement with this notion, a glycosylphosphatidylinositol-anchored protein reporter mislocalizes in osw2Δ spores. The osw2Δ mutation also exhibited a severe synthetic sporulation defect when combined with the deletion of a ß-1,6-glucan synthesis-related gene, BIG1. Osw2 is localized to the prospore membrane during sporulation. However, it disappears in mature spores, indicating that it is not a structural component of the spore wall. Given that Osw2 contains a probable 2-dehydropantoate 2-reductase domain, it may mediate an enzymatic reaction. Osw2 shows a weak similarity to other 2-dehydropantoate 2-reductase domain-containing proteins, Svl3 and Pam1. A pam1Δsvl3Δ mutant exhibits vegetative cell and spore wall defects. Thus, the 2-dehydropantoate 2-reductase domain-containing proteins may have a similar function in glucan and/or mannan layer assembly.

ß-1,6-glucan synthesis-associated genes are required for proper spore wall formation in Saccharomyces cerevisiae.

The yeast spore wall is an excellent model to study the assembly of an extracellular macromolecule structure. In the present study, mutants defective in ß-1,6-glucan synthesis, including kre1∆, kre6∆, kre9∆ and big1∆, were sporulated to analyse the effect of ß-1,6-glucan defects on the spore wall. Except for kre6∆, these mutant spores were sensitive to treatment with ether, suggesting that the mutations perturb the integrity of the spore wall. Morphologically, the mutant spores were indistinguishable from wild-type spores. They lacked significant sporulation defects partly because the chitosan layer, which covers the glucan layer, compensated for the damage. The proof for this model was obtained from the effect of the additional deletion of CHS3 that resulted in the absence of the chitosan layer. Among the double mutants, the most severe spore wall deficiency was observed in big1∆ spores. The majority of the big1∆chs3∆ mutants failed to form visible spores at a higher temperature. Given that the big1∆ mutation caused a failure to attach a GPI-anchored reporter, Cwp2-GFP, to the spore wall, ß-1,6-glucan is involved in tethering of GPI-anchored proteins in the spore wall as well as in the vegetative cell wall. Thus, ß-1,6-glucan is required for proper organization of the spore wall. Copyright © 2017 John Wiley & Sons, Ltd.

Decreased MiR-155 Level in the Peripheral Blood of Non-Alcoholic Fatty Liver Disease Patients may Serve as a Biomarker and may Influence LXR Activity.

BACKGROUND: Obesity is now a common risk factor for non-alcoholic fatty liver disease (NAFLD). Thus, it is important to explore its underlying mechanisms. METHODS: Total RNA was extracted from peripheral whole blood samples from 50 NAFLD patients and 50 healthy controls. In addition, human liver specimens were obtained through liver biopsies from NAFLD patients and healthy controls. The level of miRNA was studied using real-time PCR. The expression of lipogenic genes was analyzed using western blot, and a dual luciferase reporter assay was conducted to identify the possible target gene. Adenovirus vectors were injected into the tail vein of the high fat diet (HFD)-fed mice to study the role of miR-155 on lipid accumulation in vivo. RESULTS: The level of miR-155 was markedly reduced in the livers and peripheral blood of NAFLD patients compared with healthy controls. Upregulation of miR-155 decreased intracellular lipid content and the SREBP1 and FAS protein levels, while inhibition of miR-155 enhanced the intracellular lipid content. The dual luciferase reporter assay showed that Liver X receptor (LXR)α was the target gene of miR-155, and silencing miR-155 reduced the expression of SREBP1 and FAS. An in vivo study showed that upregulation of miR-155 decreased the hepatic lipid accumulation mainly by suppressing the LXRα-dependent lipogenic signaling pathway. CONCLUSIONS: In summary, decreased expression of miR-155 in the peripheral blood may be utilized as a potential novel biomarker for NAFLD screening mainly by targeting LXRα.

Pseudogene PTENP1 Functions as a Competing Endogenous RNA (ceRNA) to Regulate PTEN Expression by Sponging miR-499-5p.

Increasing evidence has shown that pseudogenes can widely regulate gene expression. However, little is known about the specific role of PTENP1 and miR-499-5p in insulin resistance. The relative transcription level of PTENP1 was examined in db/db mice and high fat diet (HFD)-fed mice by real-time PCR. To explore the effect of PTENP1 on insulin resistance, adenovirus overexpressing or inhibiting vectors were injected through the tail vein. Bioinformatics predictions and a luciferase reporter assay were used to explore the interaction between PTENP1 and miR-499-5p. The relative transcription level of PTENP1 was largely enhanced in db/db mice and HFD-fed mice. Furthermore, the overexpression of PTENP1 resulted in impaired Akt/GSK activation as well as glycogen synthesis, while PTENP1 inhibition led to the improved activation of Akt/GSK and enhanced glycogen contents. More importantly, PTENP1 could directly bind miR-499-5p, thereby becoming a sink for miR-499-5p. PTENP1 overexpression results in the impairment of the insulin-signaling pathway and may function as a competing endogenous RNA for miR-499-5p, thereby contributing to insulin resistance.

[Moxibustion Improved Transcutaneous Oxygen Tension and Exercise Capacity in Lower Limbs of Peripheral Arterial Disease].

OBJECTIVE: To observe the effects of moxibustion and treadmill exercise on transcutaneous oxygen tension and exercise capacity in lower limbs of peripheral arterial disease (PAD). METHODS: Totally 58 mild-to-moderate PAD patients were assigned to the control group (18 cases), the moxibustion group (20 cases), and the treadmill exercise group (20 cases) by random digit table. Patients in the control group received conventional drug therapy for 12 weeks. Patients in the moxibustion group and the treadmill exercise group additionally received moxibustion [at Zusanli (ST36), Sanyinjiao (SP6), Yongquan (KI1)] and treadmill exercise respectively, once per day, 5 times per week for 12 weeks in total. Ankle-Brachial Index (ABI) , transcutaneous oxygen tension (TcPO2), 6-min walking test (6MWT), and walking impairment questionnaire (WIQ) were assessed before and after treatment. RESULTS: Compared with the control group and the same group before treatment, there was no statistical difference in ABI in the moxibustion group and the treadmill exercise group (P > 0.05). But TcPO2, 6MWT, and WIQ were obviously elevated (P < 0.01). Besides, 6MWT and WIQ assessment of the treadmill exercise group were better than that of the moxibustion group (P < 0.01) after intervention. CONCLUSION: Moxibustion and treadmill exercise could improve the exercise capacity and TcPO2of lower limbers in PAD patients.

MiR-499-5p Contributes to Hepatic Insulin Resistance by Suppressing PTEN.

BACKGROUND: Type 2 diabetes afflicts 95% of diabetes patients. Recent data suggest that miRNAs play a key role in insulin production, secretion and function. This study aims to explore the specific role of miR-499-5p in hepatic insulin resistance. METHODS: The miRNA expression levels in the livers of db/db mice were analyzed using miRNA chips and were verified by real-time PCR. miR-499-5p mimics and an inhibitor were transfected into NCTC1469 cells. Then, the PI3K/AKT signaling pathway and glycogen level were determined. The target genes of miR-499-5p were predicted by bioinformatics and then confirmed by dual luciferase reporter assay and Western blot. To establish an insulin resistance (IR) animal model, C57BL/6 mice were fed a high-fat diet (HFD). The level of miR-499-5p in the livers of HFD-fed mice was upregulated through tail vein injection of adenovirus vectors (ad) containing miR-499-5p mimics. The glucose tolerance test (GTT) and insulin tolerance test (ITT) were used to determine glucose tolerance and insulin tolerance, respectively. RESULTS: MicroRNA chips and qPCR showed that miR-499-5p was significantly decreased in the livers of db/db mice. Downregulation of miR-499-5p impaired the insulin signaling pathway and glycogen synthesis, whereas upregulation of miR-499-5p promoted the insulin signaling pathway and glycogen synthesis in NCTC1469 cells. The dual luciferase reporter assay and Western blot demonstrated that PTEN was the target gene of miR-499-5p. Compared with the control group, miR-499-5p was increased 2.1-fold in the livers of HFD-fed mice. By tail vein injection of adenovirus vector containing miR-499-5p mimics, GTT and ITT were improved in HFD-fed mice. CONCLUSION: Downregulation of the miR-499-5p level impaired the PI3K/AKT/GSK signaling pathway and glycogen synthesis by targeting PTEN.

[Effects of unblocking meridians and Du-channel massage combined with rehabilitation training on the motor function improvement of cerebral ischemic stroke patients].

OBJECTIVE: To study effects of unblocking meridians and Du-channel massage combined with rehabilitation training on the motor function improvement of cerebral ischemic stroke (CIS) patients. METHODS: Eighty CIS patients were assigned to the treatment group (40 cases) and the control group (40 cases) using random digit table method. Bobath technique was mainly carried out in patients in the control group. On the basis of routine rehabilitation training, unblocking meridians and Du-channel massage was additionally given to patients in the treatment group. The therapeutic course for all patients was 8 weeks. The changes of Fugl-Meyer Assessment-L (FMA-L), 10 meter maximal walking speed (MWS), and gait space and time parameters were compared before and after treatment. RESULTS: Before treatment there was no significant difference in each index between the two groups (P > 0.05). After treatment there was significant difference in these indices between the two groups (P < 0.05), with better results obtained in the treatment group (P < 0.01, P < 0.05). CONCLUSION: The unblocking meridians and Du-channel massage could improve the motor function and walking capability of CIS patients, which was worthy of further summaries.