Optimization of the Preparation Process of Glucuronomannan Oligosaccharides and Their Effects on the Gut Microbiota in MPTP-Induced PD Model Mice.

Parkinson's disease (PD) is a prevalent neurodegenerative disorder, and accumulating evidence suggests a link between dysbiosis of the gut microbiota and the onset and progression of PD. In our previous investigations, we discovered that intraperitoneal administration of glucuronomannan oligosaccharides (GMn) derived from Saccharina japonica exhibited neuroprotective effects in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mouse model. However, the complicated preparation process, difficulties in isolation, and remarkably low yield have constrained further exploration of GMn. In this study, we optimized the degradation conditions in the preparation process of GMn through orthogonal experiments. Subsequently, an MPTP-induced PD model was established, followed by oral administration of GMn. Through a stepwise optimization, we successfully increased the yield of GMn, separated from crude fucoidan, from 1~2/10,000 to 4~8/1000 and indicated the effects on the amelioration of MPTP-induced motor deficits, preservation of dopamine neurons, and elevation in striatal neurotransmitter levels. Importantly, GMn mitigated gut microbiota dysbiosis induced by MPTP in mice. In particular, GM2 significantly reduced the levels of Akkermansia, Verrucomicrobiota, and Lactobacillus, while promoting the abundance of Roseburia and Prevotella compared to the model group. These findings suggest that GM2 can potentially suppress PD by modulating the gut microbiota, providing a foundation for the development of a novel and effective anti-PD marine drug.

Fucoidan from the cell wall of Silvetia siliquosa with immunomodulatory effect on RAW 264.7 cells.

Silvetia siliquosa, the only species of the family Fucaceae in China, is used as a medicine food homology. Fucoidan from S. siliquosa was extracted by hot water twice thoroughly (13 % of total yield), and a purified fucoidan SSF with a molecular weight of 93 kD was obtained. Chemical composition analysis demonstrated that SSF was primarily composed of sulfate (21.68 wt%) and fucose (84 % of all neutral monosaccharides). IR, methylation analysis, NMR and ESI-MS results indicated SSF had the backbone of mainly (1 â†’ 3)-α-L-fucopyranose and minor (1 â†’ 4)-α-L-fucopyranose, with little 1,3 and 1,4 branched ß-D-Xylp and ß-D-Galp. The in vitro immunomodulatory test on RAW 264.7 cells showed that SSF could up-regulate the expression of immune related factors and proteins in a concentration-dependent manner, but the immunomodulatory effect disappeared from desulfated SSF. This research indicated that highly sulfated fucan possessed immunomodulatory effect and the importance of sulfate groups in the activity of SSF.

Low molecular weight fucoidan LF2 improves the immunosuppressive tumor microenvironment and enhances the anti-pancreatic cancer activity of oxaliplatin.

Chemotherapy remains the cornerstone of pancreatic cancer treatment. However, the dense interstitial and immunosuppressive microenvironment frequently render the ineffective anti-tumor activity of chemotherapeutic agents. Macrophages play a key role in the tumor immunomodulation. In this study, we found that low molecular weight of fucoidan (LF2) directly regulated the differentiation of mononuclear macrophages into the CD86+ M1 phenotype. LF2 significantly upregulated the expressions of M1 macrophage-specific cytokines, including iNOS, IL-6, TNFα and IL-12. LF2 modulated macrophage phenotypic transformation through activation of TLR4-NFκB pathway. Furthermore, we observed that LF2 enhanced the pro-apoptotic activity of oxaliplatin (OXA) in vitro by converting macrophages to a tumoricidal M1 phenotype. Meanwhile, LF2 increased intratumoral M1 macrophage infiltration and ameliorated the immunosuppressed tumor microenvironment, which in turn enhanced the anti-pancreatic ductal adenocarcinoma (PDAC) activity of OXA in vivo. Taken together, our results suggested that LF2 could act as a TLR4 agonist targeting macrophages and has a synergistic effect against PDAC when combined with OXA.

Low molecular weight fucoidan restores diabetic endothelial glycocalyx by targeting neuraminidase2: A new therapy target in glycocalyx shedding.

BACKGROUND AND PURPOSE: Diabetic vascular complication is a leading cause of disability and mortality in diabetes patients. Low molecular weight fucoidan (LMWF) is a promising drug candidate for vascular complications. Glycocalyx injury predates the occurrence of diabetes vascular complications. Protecting glycocalyx from degradation relieves diabetic vascular complications. LMWF has the potential to protect the diabetes endothelial glycocalyx from shedding. EXPERIMENTAL APPROACH: The protective effect of LMWF on diabetic glycocalyx damage was investigated in db/db mice and Human Umbilical Vein Endothelial Cells (HUVEC) through transmission electron microscopy and WGA labelling. The effect of LMWF on glycocalyx degrading enzymes expression was investigated. Neuraminidase2 (NEU2) overexpression/knockdown was performed in HUVECs to verify the important role of NEU2 in glycocalyx homeostasis. The interaction between NEU2 and LMWF was detected by ELISA and surface plasmon resonance analysis (SPR). KEY RESULTS: LMWF normalizes blood indexes including insulin, triglyceride, uric acid and reduces diabetes complications adverse events. LMWF alleviates diabetic endothelial glycocalyx damage in db/db mice kidney/aorta and high concentration glucose treated HUVECs. NEU2 is up-regulated in db/db mice and HUVECs with high concentration glucose. Overexpression/knockdown NEU2 results in glycocalyx shedding in HUVEC. Down-regulation and interaction of LMWF with NEU2 is a new therapy target in glycocalyx homeostasis. NEU2 was positively correlated with phosphorylated IR-ß. CONCLUSION AND IMPLICATIONS: NEU2 is an effective target for glycocalyx homeostasis and LMWF is a promising drug to alleviate vascular complications in diabetes by protecting endothelial glycocalyx.

Preparation of Multifunctional Seaweed Polysaccharides Derivatives Composite Hydrogel to Protect Ultraviolet B-Induced Photoaging In Vitro and In Vivo.

Seaweed polysaccharides can be used for protective skin photoaging which is caused by long-term exposure to ultraviolet B (UVB). In this study, a multifunctional composite hydrogel (FACP5) is prepared using sulfated galactofucan polysaccharides, alginate oligosaccharides as active ingredients, and polyacrylonitrile modified κ-Carrageenan as substrate. The properties of FACP5 show that it has good water retention, spreadability, and adhesion. The antiphotoaging activity is evaluated in vitro and in vivo. In vitro experiments demonstrate that the components of FACP5 exhibit good biocompatibility, antioxidant, and anti-tyrosinase activities, and could reduce the cell death rate induced by UVB. In vivo experiments demonstrate that, compared with the mice skin in model group, the skin water content treated with FACP5 increases by 29.80%; the thicknesses of epidermis and dermis decrease by 53.56% and 43.98%, respectively; the activities of catalase and superoxide dismutase increase by 1.59 and 0.72 times, respectively; the contents of interleukin-6 and tumor necrosis factor-α decrease by 19.21% and 17.85%, respectively; hydroxyproline content increases by 32.42%; the expression level of matrix metalloproteinase-3 downregulates by 42.80%. These results indicate that FACP5 has skin barrier repairing, antioxidant, anti-inflammatory, and inhibiting collagen degradation activies, FACP5 can be used as a skin protection remedy for photoaging.

Fucoidan may treat jellyfish dermatitis by inhibiting the inflammatory effect of jellyfish venom.

Jellyfish dermatitis is a common medical problem caused by jellyfish stings. However, there are no targeted and effective medications for their treatment. Here, the biological activity of fucoidan for treatment of jellyfish dermatitis was investigated for the first time. 3 mg/mL Fucoidan attenuated the inflammatory effects of Nemopilema nomurai nematocyst venom (NnNV), including dermal toxicity and myotoxicity. Fucoidan may decrease the inflammatory effects of NnNV by downregulating MAPK and NF-κB pathways. This may be attributed to the inhibitory effect of fucoidan on metalloproteinases and phospholipase A2 (PLA2) in NnNV. 3 mg/mL fucoidan reduced the metalloproteinase activity in NnNV from 316.33 ± 20.84 U/mg to 177.33 ± 25.36 U/mg, while the inhibition of PLA2 activity in NnNV by 1 mg/mL fucoidan could reach 37.67 ± 3.42 %. Besides, external application of 3 mg/mL fucoidan can effectively alleviate the symptoms of jellyfish dermatitis. These observations suggest that fucoidan has considerable potential for treatment of jellyfish dermatitis and could be regarded as a novel medicine for jellyfish envenomation. This study provides new ideas for treatment of jellyfish envenomation and suggests evidence for the use of fucoidan in the treatment of jellyfish dermatitis as well as broadens the potential application of fucoidan in clinical practice.

Viscoelasticity in trapezius myofascial pain syndrome: quantitative assessment using Real-Time Shear-Wave Elastography.

OBJECTIVE: To investigate the differences in the viscoelastic properties between normal trapezius muscles and those in patients with trapezius myofascial pain syndrome (MPS) using real-time shear-wave elastography (SWE). MATERIALS AND METHODS: This study included 31 patients with trapezius MPS and 31 volunteers. Sixty-one trapezius muscles (41 and 20 on the affected and non-affected side, respectively) of patients with MPS and 62 normal trapezius muscles in volunteers were assessed. Conventional ultrasonic parameters, including skeletal muscle thickness, resistance index (RI), and mean shear wave velocity (SWVmean) of trapezius muscles, were obtained in the seated position with the shoulders and neck relaxed. The daily neck leaning time (unit:hours) of all participants was obtained using a questionnaire. RESULTS: Ultrasound showed no statistically significant differences in thickness or RI of the trapezius muscles of the affected and non-affected sides in MPS patients versus normal trapezius muscles (p = 0.976 and 0.106, respectively). In contrast, the SWVmean of trapezius muscles in patients with MPS was significantly higher than that of normal trapezius muscles in both the affected and non-affected sides (4.41 ± 1.02 m/s vs. 3.35 ± 0.79 m/s, p < 0.001; 4.05 ± 0.63 m/s vs. 3.35 ± 0.79 m/s, p = 0.002). There was no significant difference between the SWVmean of the trapezius muscles on the affected and non-affected sides in patients with MPS (4.41 ± 1.02 m/s vs. 4.05 ± 0.63 m/s, p = 0.225). Correlation analysis showed that daily neck forward time was positively correlated with the SWVmean of the trapezius muscles on the affected and non-affected sides in patients with MPS (r = 0.635, p < 0.001; r = 0.576, p = 0.008). CONCLUSION: SWE can quantitatively evaluate stiffness of trapezius muscles in patients with trapezius MPS. The stiffness of both affected and non-affected trapezius muscles increased in patients with trapezius MPS, and the degree of increase positively correlated with the time of cervical forward leaning.

Preparation of a Dual-Functional Sulfated Galactofucan Polysaccharide/Poly(vinyl alcohol) Hydrogel to Promote Macrophage Recruitment and Angiogenic Potential in Diabetic Wound Healing.

A diabetic foot ulcer is a common high-risk complication in diabetic patients, but there is still no universal dressing for clinical treatment. In this study, a novel dual-functional sulfated galactofucan polysaccharide/poly(vinyl alcohol) hydrogel (DPH20) is developed during freeze-thaw cycles. Experimental results indicated that DPH20 had a high specific surface area, a dense porous structure, and a good swelling property, which could effectively adsorb the exudates and keep the wound moist. Furthermore, DPH20 exhibited remarkably recruited macrophage capability and accelerated the inflammation stage by improving the expression of the mRNA of CCL2, CCR2, and CCL22 in macrophages. DPH20 could promote cell migration and growth factor release to accelerate tube formation under hyperglycemic conditions in cell models of L929s and HUEVCs, respectively. Significantly, DPH20 accelerates the reconstruction of the full-thickness skin wound by accelerating the recruitment of macrophages, promoting angiogenesis, and releasing the growth factor in the diabetic mouse model. Collectively, DPH20 is a promising multifunctional dressing to reshape the damaged tissue environment and accelerate wound healing. This study provides an efficient strategy to repair and regenerate diabetic skin ulcers.

Upper trapezius muscle elasticity in cervical myofascial pain syndrome measured using real-time ultrasound shear-wave elastography.

Background: Myofascial pain syndrome (MPS) is a common cause of neck pain, which is a global public health problem. Because MPS does not present morphological changes within lesioned muscles, there are no imaging diagnostic criteria for this condition. In this study, we evaluate elasticity changes in upper trapezius muscles most frequently involved in cervical MPS using real-time ultrasound shear-wave elastography, and we examine their potential diagnostic value. Methods: We consecutively enrolled 109 right posterior neck pain patients for this prospective study. Of these, 51 were diagnosed with MPS and 58 with non-MPS in the right side of their neck. Among MPS patients, 19 fell into the 1-3 range (mild pain) for pain scores on the visual analog scale (VAS), 25 fell into the 4-6 range (moderate pain), and 7 into the 7-10 range (severe pain). MPS was diagnosed by two independent clinicians using the diagnostic criteria proposed by Simons et al. Using real-time ultrasound shear-wave elastography, we measured right trapezius mean shear-wave velocity (SWVmean). The midpoint of the line between the foramen magnum and the end of the right acromion served as measuring point. Regions of interest were scaled to span 0-8.0 m/s. Results: Trapezius SWVmean was significantly higher in MPS patients compared with non-MPS patients (P<0.001). Stratified analysis of MPS patients according to pain severity revealed similar trapezius SWVmean between mild pain and non-MPS patients (P=0.324), however SWVmean was higher in moderate and severe pain MPS patients compared with non-MPS patients (P<0.001). The area under the curve (AUC) value for upper trapezius SWVmean in MPS patients was 0.791 (95% CI: 0.703-0.863). Corresponding sensitivity and specificity values were 86.27% (95% CI: 73.7-94.3%) and 62.07% (95% CI: 48.4-74.5%). Stratified analysis of MPS patients by pain severity produced the following AUC values for trapezius SWVmean in MPS patients with mild, moderate, and severe pain: 0.578 (95% CI: 0.460-0.690), 0.899 (95% CI: 0.814-0.955), and 0.983 (95% CI: 0.914-0.999), respectively. Conclusions: Elasticity changes and increased stiffness in the trapezius occur in cervical MPS patients with moderate and severe pain. The SWVmean parameter reflecting trapezius muscle elasticity may be valuable for successful screening of cervical MPS, especially in patients with moderate and severe pain.

A single-center retrospective study of the COCO technique in the treatment of chronic internal carotid artery occlusion.

OBJECTIVE: To introduce a novel endovascular recanalization method and to investigate its success rate, periprocedural complications, and early outcomes in patients with chronic internal carotid artery occlusion (CICAO). As this novel technique was designed to treat CICAO with a full coaxial system, we named it the COCO technique. METHODS: Data from consecutive patients with symptomatic CICAO who underwent endovascular recanalization in our institution were retrospectively reviewed. The COCO technique allows extracranial angioplasty and stenting with occasional intracranial angioplasty and stenting as needed to be performed in a coaxial fashion. Patients' demographic and clinical information, morphologic characteristics, procedural results, complications, and follow-up outcomes were recorded. RESULTS: Forty-nine patients were enrolled in this study. The technical success rate was 89.8% (44/49). Four patients experienced intraoperative complications, two patients had a slight subarachnoid hemorrhage, and two patients had asymptomatic dissection. Distal embolization or carotid-cavernous arteriovenous fistula was not detected. In addition, three patients developed hemorrhagic complications and three developed postoperative ischemic complications. All these patients improved after conservative treatment and subsequent rehabilitation. During the median 6 (3-6) months of follow-up, one patient died of severe pneumonia and two patients experienced recurrent ischemic events. In patients with successful recanalization, modified Rankin Scale scores were lower at the 3-month follow-up than at baseline (1 (0-2) vs 2 (1-2), P=0.04). Restenosis was observed in six (15.8%) patients. CONCLUSIONS: Our study showed that the COCO technique is effective and safe for endovascular recanalization in patients with CICAO and has low periprocedural complications and favorable functional outcomes.

Isolation, Characterization, and Anti-Idiopathic Pulmonary Fibrosis Activity of a Fucoidan from Costaria costata.

Pulmonary fibrosis is a chronic, progressive, and fatal disease of the interstitial lung. There is currently a lack of efficient therapy to reverse the prognosis of patients. In this study, a fucoidan from Costaria costata was isolated, and its anti-idiopathic fibrosis activity was investigated both in vitro and in vivo. The chemical composition analysis showed that C. costata polysaccharide (CCP) consists of galactose and fucose as the main monosaccharides with a sulfate group content of 18.54%. Further study found that CCP could resist TGF-ß1-induced epithelial-mesenchymal transition (EMT) in A549 cells by inhibiting the TGF-ß/Smad and PI3K/AKT/mTOR signaling pathways. Moreover, in vivo study found that CCP treatment alleviated bleomycin (BLM)-stimulated fibrosis and inflammation in mice lung tissue. In conclusion, the present study suggests that CCP could protect the lung from fibrosis by relieving the EMT process and inflammation in lung cells.

Effects of different dehydration methods on the preservation of aortic and renal glycocalyx structures in mice.

Glycocalyx is located outside the vascular endothelial cells playing an important role in vascular homeostasis. However, lacking efficient detection methods is one of the biggest obstacles to study the glycocalyx. In this study, three dehydration methods were used to compare the preservation of HUVEC, aorta and kidney glycocalyx by transmission electron microscope. The chemical pre-fixation was performed by lanthanum nitrate staining, and the mice aorta and renal glycocalyx were prepared by different dehydration methods such as ethanol gradient, acetone gradient and low temperature dehydration. HUVEC glycocalyx was prepared by acetone gradient and low temperature dehydration. Low temperature dehydration method preserves HUVEC and mice aortic glycocalyx completely, which had a certain thickness and presented a needle-like structure. But for mice kidney, the acetone gradient dehydration preparation method could better preserve the glycocalyx integrity than other two methods. In conclusion, low temperature dehydration method is suitable for HUVEC and aortic glycocalyx preservation, acetone gradient dehydration method is more suitable for kidney glycocalyx preservation.

Inhibitory activity of a sulfated oligo-porphyran from Pyropia yezoensis against SARS-CoV-2.

COVID-19 caused by SARS-CoV-2 has spread around the world at an unprecedented rate. A more homogeneous oligo-porphyran with mean molecular weight of 2.1 kD, named OP145, was separated from Pyropia yezoensis. NMR analysis showed OP145 was mainly composed of →3)-ß-d-Gal-(1 â†’ 4)-α-l-Gal (6S) repeating units with few replacement of 3,6-anhydride, and the molar ratio was 1:0.85:0.11. MALDI-TOF MS revealed OP145 contained mainly tetrasulfate-oligogalactan with Dp range from 4 to 10 and with no more than two 3,6-anhydro-α-l-Gal replacement. The inhibitory activity of OP145 against SARS-CoV-2 was investigated in vitro and in silico. OP145 could bind to Spike glycoprotein (S-protein) through SPR result, and pseudovirus tests confirmed that OP145 could inhibite the infection with an EC50 of 37.52 µg/mL. Molecular docking simulated the interaction between the main component of OP145 and S-protein. All the results indicated that OP145 had the potency to treat and prevent COVID-19.

Critical Evaluation of Secondary Cancer Risk After Breast Radiation Therapy with Hybrid Radiotherapy Techniques.

Background: As hybrid radiotherapy technique can effectively balance dose distribution between targets and organs, it is necessary to evaluate the late effects related to radiotherapy. The aim of the study was to calculate and provide individual estimates of the risks for hybrid radiotherapy techniques in breast cancer patients. Methods: Whole-breast irradiation was performed in 43 breast cancer patients by using 3D conformal, intensity-modulated and hybrid techniques. The excess absolute risk (EAR), lifetime attributable risk (LAR) and normal tissue complication probability (NTCP) were calculated to estimate risks in organs. The risk variability in contralateral breast was assessed by using the patient's anatomic parameters. Results: Compared with IMRT and FinF, hybrid techniques achieved satisfactory dose distribution and comparable or lower estimated risks in organs. The LAR was estimated to be up to 0.549% for contralateral lung with advantages of tangential techniques over H-VMAT. For ipsilateral lung, the LAR was estimated to be up to 9.021%, but lower in H-VMAT and FinF without significant difference. The risk of thyroid was negligible in overall estimation. For contralateral breast, the LAR was estimated to be up to 0.865% with advantages of MH-IMRT and H-VMAT over TF-IMRT. The fraction of individual variability could be explained by using anatomic parameters of minimum breast distance (MBD) and minimum target concave angle (θMTCA). NTCP for all analyzed endpoints was significantly higher in TF-IMRT relative to FinF and hybrid techniques, while TH-IMRT and H-VMAT were presenting lower toxicity risk. However, MH-IMRT presented a higher probability of toxicity in lung. For most cases, H-VMAT demonstrated a benefit for contralateral breast, heart and lung sparing. Conclusion: The optimal treatment should be performed individually according to anatomic parameters and balances between EAR and NTCP. Individual assessment may assist in achieving optimal balances between targets and organs as well as supporting clinical decision-making processes.

Impact of improved stroke green channel process on the delay of intravenous thrombolysis in patients with acute cerebral infarction during the COVID-19 pandemic: An observational study.

Objective: This study analyzed the impact of the improved stroke green channel process on the delay of intravenous thrombolysis in patients with acute cerebral infarction under coronavirus disease 2019 (COVID-19) prevention and control measures. Methods: We included 57 patients from the stroke center of the Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine before the improvement of the stroke green channel process (March-July 2019), as well as 94 patients during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak (March-July 2020) and 68 patients during the Omicron variant outbreak (March-July 2022) after the improvement of stroke green channel process. The door-to-needle time (DNT), door-to-imaging time (DIT), and door-to-test completion time were compared among the three groups. We analyzed the impact of this process improvement in the emergency green channel during the pandemic on the delay of intravenous thrombolysis. Results: This study included a total of 229 patients with acute cerebral infarction who went through the green channel for intravenous thrombolysis (57 in the pre-pandemic group, 94 in the SARS-CoV-2 outbreak group, and 68 in the Omicron outbreak group). The percentages of patients undergoing intravenous thrombolysis in the pre-pandemic, SARS-CoV-2 outbreak, and Omicron outbreak groups differed significantly (19.32%, 22.27%, and 28.94%, respectively, P = 0.029). Compared to the pre-pandemic group, the National Institutes of Health Stroke Scale (NIHSS) score at admission was significantly higher in the Omicron outbreak group (7.71 ± 7.36 in the Omicron outbreak group vs. 5.00 ± 4.52 in the pre-pandemic group) (P = 0.026) but not in the SARS-CoV-2 outbreak group (4.79 ± 5.94 in the SARS-CoV-2 outbreak group vs. 5.00 ± 4.52 in the pre-pandemic group, P = 0.970). Significantly higher proportions of patients undergoing emergency intravenous thrombolysis came to the hospital by ambulance in the SARS-CoV-2 and Omicron outbreak groups compared to the pre-pandemic group (38.6% in the pre-pandemic group, 51.1% in the SARS-CoV-2 outbreak group, and 82.4% in the Omicron outbreak group, P < 0.001). Compared to the pre-pandemic group, the DIT was significantly higher in the SARS-CoV-2 outbreak group (22.42 ± 7.62 min in the SARS-CoV-2 outbreak group vs. 18.91 ± 8.23 min in the pre-pandemic group, P =0.031) but not the Omicron outbreak group (20.35 ± 10.38 min in the Omicron outbreak group vs. 18.91 ± 8.23 min in the pre-pandemic group, P = 0.543). The door-to-test completion time was significantly longer in the SARS-CoV-2 and Omicron outbreak groups compared to that in the pre-pandemic group (78.37 ± 25.17 min in the SARS-CoV-2 outbreak group, 92.60 ± 25.82 min in the Omicron outbreak group vs. 65.11 ± 22.35 min in the pre-pandemic group, P < 0.001); however, the DNT in the SARS-CoV-2 and Omicron outbreak groups did not differ significantly from those in the pre-pandemic group (both P > 0.05). Conclusion: During the two periods of the COVID-19 outbreak (SARS-CoV-2 and Omicron), after the improvement of the green channel for intravenous thrombolysis, there might be some delay in in-hospital DIT during the SARS-CoV-2 outbreak, however, the in-hospital delay indicator DNT for intravenous thrombolysis were not affected.

Fucoidan, as an immunostimulator promotes M1 macrophage differentiation and enhances the chemotherapeutic sensitivity of capecitabine in colon cancer.

Chemotherapy resistance is one of the most critical challenges in colorectal cancer (CRC) treatment. The occurrence and development of chemotherapy resistance closely related to the tumor immune microenvironment (TIME). As the most important immunosuppressive immune cells infiltrating into the TIME, macrophages are essential for chemotherapy resistance in CRC treatment. In this study, we found that a kind of fucoidan (FPS1M) induced macrophages differentiation to the M1 phenotype, and this transformation promoted cancer cells apoptosis both in vitro and in vivo. TNFα is a key mediator of FPS1M-induced tumorcidal activity of macrophages. Mechanistically, as a stimulator of TLR4, FPS1M enhanced macrophages glycolysis and regulated macrophages differentiation to the M1 phenotype by the activation of TLR4 mediated PI3K/AKT/mTOR signaling axis. In addition, FPS1M improved the immunosuppressed tumor microenvironment by increasing the infiltration of M1 macrophages in tumor tissue, which was conducive to improving the sensitivity of tumor to chemotherapy. Collectively, our findings demonstrated that FPS1M has the great potential to be used in tumor immunotherapy. The results also suggested that the combination of FPS1M with capecitabine is an alternative therapy method for colon cancer.

Isolation, identification and in vivo antihypertensive effect of novel angiotensin I-converting enzyme (ACE) inhibitory peptides from Spirulina protein hydrolysate.

The side effects of traditional antihypertensive drugs have driven people's interest in the discovery of novel angiotensin-I converting enzyme (ACE) inhibitory peptides with efficiency and safety. Spirulina possesses abundant proteins and is considered as a good source of bioactive peptides. To identify ACE inhibitory peptides, Spirulina protein was hydrolyzed by protease K, and the hydrolysate was separated by gel permeation chromatography and reverse phase HPLC. Thirty peptide sequences were identified from the ACE inhibitory fractions by LC-MS/MS, and 15 potential ACE inhibitory peptides were further selected by combined virtual screening and experimental investigation. Among the identified Spirulina peptides, TVLYEH (SpH-6, IC50 = 2.88 µM) and LQAGGLF (SpH-7, IC50 = 66.83 µM) were found to possess potent ACE inhibitory activity. The inhibition patterns of SpH-6 and SpH-7 were characterized as competitive inhibition. Molecular docking indicated that SpH-6 and SpH-7 could bind to the active pockets of ACE by forming hydrogen bonds, salt bridges and pi-pi stacking. Both SpH-6 and SpH-7 exhibited stable ACE inhibitory activity under different pH (2, 4, 6, 8, 10, 12) and temperature (0, 20, 40, 60, 80, 100 °C) conditions. The ACE inhibitory activity of SpH-6 decreased slightly after simulated gastrointestinal digestion, and SpH-7 was unstable against digestive enzymes. The in vivo antihypertensive effect demonstrated that oral administration of SpH-6 significantly reduced the systolic blood pressure and diastolic blood pressure of spontaneously hypertensive rats. The present study revealed the potential antihypertensive effects of SpH-6 and indicated that SpH-6 and Spirulina protein hydrolysate could be applied as nutritional supplements for blood pressure control.

Discovery of a novel jellyfish venom metalloproteinase inhibitor from secondary metabolites isolated from jellyfish-derived fungus Aspergillus versicolor SmT07.

The major jellyfish stings that occur in China are caused by scyphozoan Nemopilema nomurai, whose venom exhibits significant metalloproteinase activity that contributes to the toxic effects of jellyfish envenomation. Researching effective inhibitors suppressing the metalloproteinase activity of jellyfish venom represents a new attempt to cure jellyfish envenomations. In the present study, secondary metabolites produced by the jellyfish-associated fungus Aspergillus versicolor SmT07 were isolated and evaluated for their anti-proteolytic activities. Two xanthones, sterigmatocystin (JC-01) and oxisterigmatocystin C (JC-06), and four alkaloids, cottoquinazoline A (JC-02), phenazine-1-carboxylic acid (JC-03), viridicatin (JC-04) and viridicatol (JC-05), were isolated and identified. Only phenazine-1-carboxylic acid (PCA) showed significant anti-proteolytic activity of jellyfish venom assayed on azocasein, and the IC50 value was 2.16 mM. PCA also significantly inhibited fibrinogenolytic activity, protecting the Bß chain of fibrinogen from degradation when preincubated with jellyfish venom at a ratio of >1:0.6 (PCA:venom, w/w). Molecular docking with several well-characterized snake venom metalloproteinases suggested the venom metalloproteinases inhibitory property of PCA by forming complex interactions with the active site via hydrogen bonds, π-π stacking and salt bridges, which was distinct from the binding mode of batimastat. The present study represents the first study identifying natural jellyfish venom metalloproteinase inhibitors from marine natural products, which may provide an alternative to develop therapeutic agents for treating jellyfish envenomations.

Association between blood pressure variability and clinical outcomes after successful recanalization in patients with large vessel occlusion stroke after mechanical thrombectomy.

Objective: Nearly half of patients who undergo mechanical thrombectomy (MT) do not experience a favorable outcome. The association between blood pressure fluctuation and clinical outcomes after successful MT is controversial. We evaluated the influence of blood pressure variability (BPV) on the clinical outcomes of stroke patients with large vessel occlusion (LVO) who underwent successful recanalization after MT. Methods: Patients with anterior circulation LVO stroke who underwent successful emergency MT (modified Thrombolysis in Cerebral Infarction, mTICI ≥ 2b) at the Shanghai Tenth People's Hospital of Tongji University from 2017 to 2021 were enrolled. Multivariate logistic models were used to investigate the association between BPV (mean arterial pressure [MAP] assessed using the standard deviation [SD]) and clinical outcomes. The primary outcome was 90-day modified Rankin Scale scores (mRS), and the secondary outcomes were 30-day mortality and symptomatic intracranial hemorrhage (sICH). Results: A total of 458 patients (56.8% men), with a mean age of 72 ± 1 years, were enrolled. Among them, 207 (45.2%) patients had unfavorable functional outcomes (mRS score 3-6) at 90 days, 61 (13.3%) patients died within 30 days, and 20 (4.4%) patients had sICH. In a fully adjusted model, BPV was associated with a higher risk of a 90-day mRS score of 3-6 (P = 0.04), 30-day mortality (P < 0.01), and sICH (P < 0.01). A significant interaction between MAP SD and rescue futile recanalization treatment was observed (P < 0.01). Conclusions: Among patients with LVO stroke who underwent successful recanalization, higher BPV was associated with worse functional outcomes, especially in those who underwent rescue treatment.

Functional Liver Imaging in Radiotherapy for Liver Cancer: A Systematic Review and Meta-Analysis.

Backgrounds: Functional liver imaging can identify functional liver distribution heterogeneity and integrate it into radiotherapy planning. The feasibility and clinical benefit of functional liver-sparing radiotherapy planning are currently unknown. Methods: A comprehensive search of several primary databases was performed to identify studies that met the inclusion criteria. The primary objective of this study was to evaluate the dosimetric and clinical benefits of functional liver-sparing planning radiotherapy. Secondary objectives were to assess the ability of functional imaging to predict the risk of radiation-induced liver toxicity (RILT), and the dose-response relationship after radiotherapy. Results: A total of 20 publications were enrolled in descriptive tables and meta-analysis. The meta-analysis found that mean functional liver dose (f-MLD) was reduced by 1.0 Gy [95%CI: (-0.13, 2.13)], standard mean differences (SMD) of functional liver volume receiving ≥20 Gy (fV20) decreased by 0.25 [95%CI: (-0.14, 0.65)] when planning was optimized to sparing functional liver (P >0.05). Seven clinical prospective studies reported functional liver-sparing planning-guided radiotherapy leads to a low incidence of RILD, and the single rate meta-analysis showed that the RILD (defined as CTP score increase ≥2) incidence was 0.04 [95%CI: (0.00, 0.11), P <0.05]. Four studies showed that functional liver imaging had a higher value to predict RILT than conventional anatomical CT. Four studies established dose-response relationships in functional liver imaging after radiotherapy. Conclusion: Although functional imaging modalities and definitions are heterogeneous between studies, but incorporation into radiotherapy procedures for liver cancer patients may provide clinical benefits. Further validation in randomized clinical trials will be required in the future.