Cross­talk between lymphangiogenesis and malignant melanoma cells: New opinions on tumour drainage and immunization (Review).

Malignant melanoma (MM) is a highly aggressive tumour that can easily metastasize through the lymphatic system at the early stages. Lymph node (LN) involvement and lymphatic vessel (LV) density (LVD) represent a harbinger of an adverse prognosis, indicating a strong link between the state of the lymphatic system and the advancement of MM. Permeable capillary lymphatic vessels are the optimal conduits for melanoma cell (MMC) invasion, and lymphatic endothelial cells (LECs) can also release a variety of chemokines that actively attract MMCs expressing chemokine ligands through a gradient orientation. Moreover, due to the lower oxidative stress environment in the lymph compared with the blood circulation, MMCs are more likely to survive and colonize. The number of LVs surrounding MM is associated with tumour-infiltrating lymphocytes (TILs), which is crucial for the effectiveness of immunotherapy. On the other hand, MMCs can release various endothelial growth factors such as VEGF-C/D-VEGFR3 to mediate LN education and promote lymphangiogenesis. Tumour-derived extracellular vesicles are also used to promote lymphangiogenesis and create a microenvironment that is more conducive to tumour progression. MM is surrounded by a large number of lymphocytes. However, both LECs and MMCs are highly plastic, playing multiple roles in evading immune surveillance. They achieve this by expressing inhibitory ligands or reducing antigen recognition. In recent years, tertiary lymphoid structures have been shown to be associated with response to anti-immune checkpoint therapy, which is often a positive prognostic feature in MM. The present review discusses the interaction between lymphangiogenesis and MM metastasis, and it was concluded that the relationship between LVD and TILs and patient prognosis is analogous to a dynamically tilted scale.

Risk factor mining and prediction of urine protein progression in chronic kidney disease: a machine learning- based study.

BACKGROUND: Chronic kidney disease (CKD) is a global public health concern. Therefore, to provide timely intervention for non-hospitalized high-risk patients and rationally allocate limited clinical resources is important to mine the key factors when designing a CKD prediction model. METHODS: This study included data from 1,358 patients with CKD pathologically confirmed during the period from December 2017 to September 2020 at Zhongshan Hospital. A CKD prediction interpretation framework based on machine learning was proposed. From among 100 variables, 17 were selected for the model construction through a recursive feature elimination with logistic regression feature screening. Several machine learning classifiers, including extreme gradient boosting, gaussian-based naive bayes, a neural network, ridge regression, and linear model logistic regression (LR), were trained, and an ensemble model was developed to predict 24-hour urine protein. The detailed relationship between the risk of CKD progression and these predictors was determined using a global interpretation. A patient-specific analysis was conducted using a local interpretation. RESULTS: The results showed that LR achieved the best performance, with an area under the curve (AUC) of 0.850 in a single machine learning model. The ensemble model constructed using the voting integration method further improved the AUC to 0.856. The major predictors of moderate-to-severe severity included lower levels of 25-OH-vitamin, albumin, transferrin in males, and higher levels of cystatin C. CONCLUSIONS: Compared with the clinical single kidney function evaluation indicators (eGFR, Scr), the machine learning model proposed in this study improved the prediction accuracy of CKD progression by 17.6% and 24.6%, respectively, and the AUC was improved by 0.250 and 0.236, respectively. Our framework can achieve a good predictive interpretation and provide effective clinical decision support.

Angiopoietin-2 expression and its relationship with lymphangiogenesis and clinicopathological characteristics in cutaneous malignant melanoma.

Objective: The aim of this study was to investigate angiopoietin-2 (Ang-2/ANGPT2) expression and its relationship with lymphangiogenesis and clinicopathological characteristics in cutaneous malignant melanoma (CMM). Methods: Gene expression differences between metastatic melanoma and melanoma in situ in 472 patients from the TCGA database were analyzed. The target gene Ang-2 was screened. A clinical study was conducted to analyze the correlation between Ang-2 expression in CMM and tumor-associated lymphangiogenesis. A total of 42 patients with primary CMM who underwent extended tumor resection procedures at the Affiliated Hospital of Jiangsu University were included in this study. Clinical data (gender, age, lymph node metastasis, Breslow thickness, and clinical stage) were collected. The expression levels of both Ang-2 and lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) proteins were detected by immunohistochemistry (IHC). Lymphatic vascular density (LVD) was counted by using LYVE-1 to label lymphatic endothelial cells (LECs) in peritumoral and intratumoral areas per high-magnification field of view. Statistical analysis was performed using the Pearson correlation test and Student's t-test. Results: Using the TCGA database, it was found that the gene expression level of Ang-2 in 368 cases of metastatic melanoma was significantly higher than that in 104 cases of melanoma in situ. Correlation analysis showed a significant relationship between Ang-2 and LYVE-1, and vascular endothelial growth factor receptor 3(VEGFR3) expression, respectively, in CMM. Moreover, the optimal cutoff value of survival analysis showed that high Ang-2 expression in CMM had a worse prognosis, based on data from the TCGA database. Our research showed that Ang-2 was more highly expressed in the group of patients with lymph node metastasis and in the group of stage 3C-4 patients than in the group of patients with no lymph node metastasis and in the group of stage 0-3B patients. Our research also showed that LVD in the group of patients with lymph node metastasis and in the group of stage 3C-4 patients was significantly higher than that in the group of no lymph node metastasis and in the group of stage 0-3B patients, respectively. Breslow thickness also correlated with Ang-2 expression and LVD. Ang-2 expression was not related to sex or age. Ang-2 expression was obviously correlated with LVD. Conclusion: An evaluation of Ang-2 expression and LVD can be used to predict the risk of tumor lymphatic metastasis and determine the prognosis of CMM. These results may also provide a new clinical treatment strategy for CMM.

Acid-sensing ion channel 1a exacerbates renal ischemia-reperfusion injury through the NF-κB/NLRP3 inflammasome pathway.

Ischemia-reperfusion injury (IRI) is the main cause of acute kidney injury (AKI), and there is no effective therapy. Microenvironmental acidification is generally observed in ischemic tissues. Acid-sensing ion channel 1a (ASIC1a) can be activated by a decrease in extracellular pH which mediates neuronal IRI. Our previous study demonstrated that, ASIC1a inhibition alleviates renal IRI. However, the underlying mechanisms have not been fully elucidated. In this study, we determined that renal tubule-specific deletion of ASIC1a in mice (ASIC1afl/fl/CDH16cre) attenuated renal IRI, and reduced the expression of NLRP3, ASC, cleaved-caspase-1, GSDMD-N, and IL-1ß. Consistent with these in vivo results, inhibition of ASIC1a by the specific inhibitor PcTx-1 protected HK-2 cells from hypoxia/reoxygenation (H/R) injury, and suppressed H/R-induced NLRP3 inflammasome activation. Mechanistically, the activation of ASIC1a by either IRI or H/R induced the phosphorylation of NF-κB p65, which translocates to the nucleus and promotes the transcription of NLRP3 and pro-IL-1ß. Blocking NF-κB by treatment with BAY 11-7082 validated the roles of H/R and acidosis in NLRP3 inflammasome activation. This further confirmed that ASIC1a promotes NLRP3 inflammasome activation, which requires the NF-κB pathway. In conclusion, our study suggests that ASIC1a contributes to renal IRI by affecting the NF-κB/NLRP3 inflammasome pathway. Therefore, ASIC1a may be a potential therapeutic target for AKI. KEY MESSAGES: Knockout of ASIC1a attenuated renal ischemia-reperfusion injury. ASIC1a promoted the NF-κB pathway and NLRP3 inflammasome activation. Inhibition of the NF-κB mitigated the NLRP3 inflammasome activation induced by ASIC1a.

A dual-task-embedded virtual reality system for intelligent quantitative assessment of cognitive processing speed.

Introduction: Processing speed is defined as the ability to quickly process information, which is generally considered as one of the affected cognitive functions of multiple sclerosis and schizophrenia. Paper-pencil type tests are traditionally used in the assessment of processing speed. However, these tests generally need to be conducted under the guidance of clinicians in a specific environment, which limits their application in cognitive assessment or training in daily life. Therefore, this paper proposed an intelligent evaluation method of processing speed to assist clinicians in diagnosis. Methods: We created an immersive virtual street embedded with Stroop task (VR-Street). The behavior and performance information was obtained by performing the dual-task of street-crossing and Stroop, and a 50-participant dataset was established with the label of standard scale. Utilizing Pearson correlation coefficient to find the relationship between the dual-task features and the cognitive test results, and an intelligent evaluation model was developed using machine learning. Results: Statistical analysis showed that all Stroop task features were correlated with cognitive test results, and some behavior features also showed correlation. The estimated results showed that the proposed method can estimate the processing speed score with an adequate accuracy (mean absolute error of 0.800, relative accuracy of 0.916 and correlation coefficient of 0.804). The combination of Stroop features and behavior features showed better performance than single task features. Discussion: The results of this work indicates that the dual-task design in this study better mobilizes participants' attention and cognitive resources, and more fully reflects participants' cognitive processing speed. The proposed method provides a new opportunity for accurate quantitative evaluation of cognitive function through virtual reality.

Pan-cancer analysis and experimental validation of DTL as a potential diagnosis, prognosis and immunotherapy biomarker.

BACKGROUND: DTL has been found to be related with multiple cancers. However, comprehensive analyses, which identify the prediction value of DTL in diagnosis, prognosis, immune infiltration and treatment, have rarely been reported so far. METHODS: Combined with the data online databases, the gene expression, gene mutation, function enrichment and the correlations with the immunity status and clinical indexes of DTL were analyzed. Expression of DTL and the degree of immune cell infiltration were examined by immunofluorescence (IF) and immunohistochemistry (IHC) and analyzed by statistical analysis. Furthermore, the influences of DTL on the cell cycle, cell proliferation and apoptosis were detected by live cell imaging, IF and flow cytometric (FC) analysis. Genomic stability assays were conducted by chromosome slide preparation. RESULTS: DTL was widely expressed in various cells and tissues, while it was overexpressed in tumor tissues except acute myeloid leukemia (LAML). Pan-cancer bioinformatics analysis showed that the expression of DTL was correlated with the prognosis, immunotherapy, and clinical indexes in various cancers. In addition, gene set enrichment analysis (GSEA) uncovered that DTL was enriched in oocyte meiosis, pyrimidine metabolism, the cell cycle, the G2M checkpoint, mTORC1 signaling and E2F targets. Furthermore, the overexpression of DTL, and its association with immune cell infiltration and clinical indexes in liver hepatocellular carcinoma (LIHC), bladder urothelial carcinoma (BLCA) and stomach adenocarcinoma (STAD) were verified in our study. It was also verified that overexpression of DTL could regulate the cell cycle, promote cell proliferation and cause genomic instability in cultured cells, which may be the reason why DTL plays a role in the occurrence, progression and treatment of cancer. CONCLUSIONS: Collectively, this study suggested that DTL is of clinical value in the diagnosis, prognosis and treatment of various cancers, and may be a potential biomarker in certain cancers.

Sebaceous Carcinoma of the Submandibular Gland a Case Report and Review of the Literature.

Objective: Sebaceous carcinoma (SC) of the submandibular gland is extremely rare. Owing to the low morbidity and nonspecific clinical manifestations, diagnosis is commonly delayed, which increases metastasis and mortality. To date, there have been five reported cases of SC of the submandibular gland. Here, we present a new case and review the relevant literature. Methods and Results: A 36-year-old woman presented with an enlarged left submandibular gland. Clinical features included a non-tender solitary nodular mass with normal overlying skin. There were no special findings on computed tomography or ultrasound examination except for a swollen mass in the left submandibular gland. The patient underwent surgical resection. Pathological examination confirmed the diagnosis of SC with nerve infiltration. Immunohistochemical examination of this case showed positive staining for P63, P40, CK7, CK8/18, MLH1, MSH2, MSH6, and PMS2. The specimen was negative for androgen receptor, CEA, S-100, CK5/6, SOX-10, SOX-11, SMA, and GCDFP-15. The KI-67 labeling index was determined to be 15%. PAS and anti-epithelial membrane antigen were positive in partial area. The patient is still undergoing follow-up, and no metastasis or recurrence has been observed for 2 months. Conclusion: This case highlighted the fact that despite its rarity, SC should be considered as a differential diagnosis for masses located in the head and face. Early and accurate diagnosis, followed by wide surgical excision, has a favorable prognosis. Therefore, clinicians should be familiar with the clinical and pathological features of this disease.

Efficacy of dexmedetomidine versus midazolam when combined with butorphanol for sedation and analgesia during burn dressing changes: A randomized clinical trial.

Objective: The aim of this study was to compare dexmedetomidine-butorphanol (DB) and midazolam-butorphanol (MB) combinations for sedation, and analgesia in burn patients undergoing dressing changes. Methods: A total of 56 ASA I-II burn patients were included in this single-center randomized clinical trial. The ages of these patients were between 20 and 60 years. TBSA ranged from 10% to 50%. They were randomized to group DB and group MB during dressing change. In the DB group, each patient received a bolus dose of dexmedetomidine (0.5 µg kg-1) and intermittent boluses of butorphanol (20 µg kg-1). In the MB group, each patient received a bolus dose of midazolam (0.05 mg kg-1) and intermittent boluses of butorphanol (20 µg kg-1). The primary outcomes were sedation scores and pain scores. The second outcomes were vital signs, side effects, and butorphanol consumption. Results: The sedation scores of these two groups did not differ significantly (p > 0.05), and the pain scores of these groups were not significantly different (p > 0.05). More patients had hypotension in the DB group than in the MB group (6 versus 0, p = 0.01), but the number of patients who had respiratory depression was higher in the MB group compared with the DB group (4 versus 0, p = 0.038). Butorphanol consumption in the MB group was higher than in the DB group (p = 0.025). Conclusion: Dexmedetomidine is comparable to midazolam when combined with butorphanol in burn patients during dressing change. Compared with midazolam, it has the advantage of opioid-sparing effect. Clinical Trial Registration: [http://www.chictr.org.cn/showproj.aspx&proj=130622], identifier [ChiCTR2100049325].

Clinical Monitoring of Serum Levels of Vitamins A, D and E in Children with Recurrent Respiratory Tract Infections of Different Ages: A Clinical Controlled Trial.

Objective: To study serum levels of vitamins A, D and E in children with recurrent respiratory tract infections of different ages and the correlation. Methods: The clinical data of two groups of children of different ages were collected. The serum levels and deficiencies of vitamins A, D and E in children were statistically analyzed. Results: The proportions of premature infants, low body weight infants, special physique, hospitalization history, hypocalcemia, living in a bungalow, and daily outdoor activities in less than 30 minutes in the case group were higher than those in the control group (χ 2=4.507, 5.165, 7.040, 14.907, 4.267, 33.800, 4.507, 8.571, P < 0.05). The serum levels of vitamins A, D and E of children aged 0-1, 2-5, and 6-12 in the case group were lower than those in the control group (P < 0.05). Compared with the control group, the serum vitamin A level of children in the case group was lower (t = 2.631, P < 0.05), and the deficiency rate was higher (χ 2=24.200, P < 0.05). Conclusion: Serum levels of vitamins A, D and E, which are related to birth mode, physical fitness, hospitalization history, hypocalcemia, vitamin deficiency, living environment, and daily outdoor activity time, vary in children with recurrent respiratory tract infections of different ages, and are lower in children with recurrent respiratory tract infections than in healthy children.

Dynamic monitoring and a clinical correlation analysis of the serum vitamin A, D, and E levels in children with recurrent respiratory tract infections.

OBJECTIVE: To investigate the correlation of the serum vitamin A, D, and E (VA, VD, and VE) levels with the occurrence and development of recurrent respiratory tract infections (RRTIs). METHODS: A total of 129 children with respiratory tract infections (RTIs) treated in our hospital from January 2018 to February 2020 (the RTIs group) and 50 healthy children undergoing physical examinations (the control group) in our hospital were recruited as the study cohort. The serum VA, VD, and VE levels were measured upon admission (the active phase) and at two weeks after discharge (the stable phase). The serum VA, VD, and VE levels in the children with RRTIs were compared with the levels in the control group, and the correlation between these three vitamins and the occurrence and development of RRTIs was analyzed. RESULTS: The RRTIs group and the RTIs group witnessed markedly lower serum VA, VD, VE, and humoral immunity index levels, including IgG, IgA, and IgM, compared to the control group, with an apparent lower outcome in the RRTIs group than in the RTIs group. The serum levels of the above indexes in the RRTIs children were reduced in the active phase compared with the stable phase. A Pearson correlation analysis showed a positive correlation between VA and IgA. A multivariate logistic regression analysis revealed that a low BMI (Body mass index), prematurity, VA deficiency, VD deficiency, and VE deficiency were the risk factors for RRTIs in children, and outdoor activity was the protective factor. CONCLUSION: The VA, VD, and VE levels are closely related to RRTIs in children. It is important to determine and supplement the VA, VD, and VE levels to prevent RTIs in children.

Probiotics combined with zinc and selenium preparation in the treatment of child rotavirus enteritis.

OBJECTIVE: To investigate clinical effect of probiotics combined with zinc and selenium preparation in the treatment of child rotavirus enteritis. METHODS: In this retrospective study, the patients were divided into two groups based on treatment method. The control group (n=42) received probiotic therapy, while the experimental group (n=43) received probiotics combined with zinc and selenium preparation. Clinical efficacy, stool frequency and incidence of adverse reactions after treatment were compared to assess the clinical effect. RESULTS: The clinical effect was improved after intervention (P<0.05), and the total effective rate of two groups was 88.4% (38/43), 50% (21/42), respectively. Time to symptom disappearance was significantly decreased in the experimental group as compared to the control group. The myocardial zymogram indices (CK, CK-MB and AST) were decreased after treatment, and the levels in the experimental group were significantly lower than in the control group (P<0.05). Moreover, we observed that the levels of inflammatory factors (IL-6, IL-8 and hsCRP) in the experimental group were significantly lower than those of the control group after intervention (all P<0.05). CONCLUSION: Probiotics combined with zinc and selenium preparation can not only significantly improve the clinical effect, but also shorten the course of disease.

Circadian rhythm disorders elevate macrophages cytokines release and promote multiple tissues/organs dysfunction in mice.

BACKGROUND: Circadian rhythm disorders are severe threats to human health. The negative impact of circadian rhythm disorders on tissues/organs has not been systematically analyzed. Therefore, there is an urgent need to evaluate the damage caused by circadian rhythm disorders and explore the possible mechanisms. METHODS: Six-week-old male mice were divided into the control (Con) group (normal circadian rhythm), L24 group (constant light), D12L12 group (weekly shift light/dark cycle), and D24 group (constant dark). Body weight was recorded every 10 days. Ninety days after model construction, the serum lipid and cytokine level, liver function, fat accumulation, carotid artery stenosis, and cardiomyopathological changes were detected in mice. Macrophages in the liver, subscapular fat, and heart tissues were labeled with immunofluorescence staining. Mouse peritoneal macrophages were then isolated. Inflammatory cytokine levels were measured in the macrophage supernatant. The ability of macrophages to form foam cells was also tested. The supernatant from macrophages in different groups was added to AML12 (hepatocytes), 3T3-L1 (preadipocytes), or HL-1 (cardiomyocytes). Effects of conditioned media on recipient cells were determined. RESULTS: Body weight, serum lipids and cytokines, subscapular fat accumulation, liver enzymes, carotid artery stenosis, and myocardial fibrosis levels of the L24, D12L12, and D24 groups mice were significantly higher than those in the Con group. Macrophages were significantly increased in the liver, heart, and subscapular fat of mice with circadian rhythmdisorders. Cytokine secretion by peritoneal macrophages was enhanced in the L24, D12L12, and D24 groups. Under oxidized low density lipoprotein (oxLDL) stimulation, macrophages with circadian rhythm disorders are more likely to form foam cells. Conditioned media from the L24, D12L12, and D24 groups significantly promoted AML12 apoptosis and lipid intake, accelerated the adipogenic differentiation of 3T3-L1, and up-regulated collagen I in HL-1. CONCLUSION: These findings reveal that macrophages are increased in the tissues/organs under circadian rhythm disorders, and these macrophages could aggravate obesity, promote liver disease, accelerate atherosclerosis, and increase myocardial fibrosis through the paracrine effect.

The correlation between the vitamin A, D, and E levels and recurrent respiratory tract infections in children of different ages.

OBJECTIVE: To investigate the correlation between the vitamin A, D, and E levels and recurrent respiratory tract infections (RRTIs) in children of different ages. METHODS: A total of 150 RRTI patients were divided into three groups: the 0 to 2 year-old group, the 3-5 year-old group, and the 6-14 year-old group. Collectively, we refer to the three groups as the RRTI group. The serum vitamin A, D and E levels were measured in the three groups. Healthy children without RRTIs were recruited as a control group. The correlations between the changes in the vitamin A, D, and E levels and the RRTIs were analyzed. RESULTS: The vitamin A, D, and E levels decreased significantly in the children with RRTIs, but only the vitamin A and D levels were negatively correlated with the incidence of RRTIs, while the vitamin E levels were not significantly correlated with the incidence of RRTIs. The follow-up results showed that the serum vitamin A, D, and E levels in the RRTI group were significantly increased after the treatment, and the WBC and CRP levels were remarkably reduced. CONCLUSION: Monitoring the serum vitamin A and D levels helps determine the disease severity, and the supplementation of adequate vitamin A and D through diet or drugs is of great help in treating RRTIs.

Butorphanol in combination with dexmedetomidine provides efficient pain management in adult burn patients.

OBJECTIVE: This study aimed to compare the sedation and analgesic effects of butorphanol alone and butorphanol in combination with dexmedetomidine on dressing changes in adult burn patients. METHOD: From June 2016 to May 2019, 44 adult burn patients from our department were enrolled in this prospective, double-blinded study. Their total burn surface area (TBSA) varied from 10% to 30%; and the depth of burn injury ranged from second degree to third degree. The patients were randomized into two groups. In the control group, butorphanol combined with saline was injected into the body via venous route during dressing change. In the observation group, butorphanol in combination with dexmedetomidine was injected. The variation in mean blood pressure, heart rate, respiratory rate, and peripheral oxygen saturation were recorded at various time-points of the procedure. Visual Analogue Scale (VAS) of pain and Ramsay Sedation Scores (RSS) were also recorded at different time points. Consumption of butorphanol and adverse events in these two groups were compared. RESULTS: The mean blood pressure and heart rate were significantly decreased in the observation group before butorphanol injection (P < 0.05) and before the dressing change (P < 0.05). The respiratory rates and peripheral oxygen saturation of these two groups showed no significant differences at all time points (P > 0.05). Patients in the observation group had lower VAS scores during dressing change (P < 0.05). The RSS Scores in the observation group were higher than those in the control group during (P < 0.05) and after the dressing change (P < 0.05). The consumption of butorphanol was more in the control group (P < 0.05), and the adverse events recorded in the control group were higher (P < 0.05). CONCLUSION: Butorphanol combined with dexmedetomidine can reduce analgesic use of butorphanol during dressing change. This combination resulted in a higher sedation score and fewer adverse effects.

Exosomes Derived from Human Umbilical Cord Mesenchymal Stem Cells Accelerate Cutaneous Wound Healing by Enhancing Angiogenesis through Delivering Angiopoietin-2.

The underlying mechanisms of human umbilical cord mesenchymal stem cells (hucMSCs) and their exosomes (hucMSC-Exs), which play significant roles in skin wound healing, remain poorly understood. By using a rat model of deep second-degree burn injury, the roles of hucMSC-Exs in angiogenesis and cutaneous wound healing in vivo were investigated. We found that hucMSC-Exs accelerated skin wound healing and angiogenesis, inducing a higher wound-closure rate and increased expression of CD31 in vivo. We also discovered that hucMSC-Exs contained angiopoietin-2 (Ang-2), and treatment with hucMSC-Exs enhanced the expression of the Ang-2 protein in the wound area and human umbilical vein endothelial cells (HUVECs) through exosomal-mediated Ang-2 transfer. Moreover, hucMSC-Exs promoted the proliferative, migratory, and tube-forming ability of HUVECs. Furthermore, overexpression of Ang-2 in hucMSC-Exs further enhanced HUVEC migration and tube formation and exerted therapeutic and proangiogenic effects in cutaneous wounds in rats, whereas knockdown of Ang-2 in hucMSC-Exs abrogated these therapeutic and proangiogenic effects. Taken together, our results indicated that hucMSC-Ex-derived Ang-2 plays a significant role in tube formation of HUVECs and promotion of angiogenesis, and further suggested that hucMSC-Ex-based therapy may serve as a promising therapeutic approach for promoting cutaneous wound healing.

Human Umbilical Cord MSC-Derived Exosomes Suppress the Development of CCl4-Induced Liver Injury through Antioxidant Effect.

Mesenchymal stem cells (MSCs) have been increasingly applied into clinical therapy. Exosomes are small (30-100 nm in diameter) membrane vesicles released by different cell types and possess the similar functions with their derived cells. Human umbilical cord MSC-derived exosomes (hucMSC-Ex) play important roles in liver repair. However, the effects and mechanisms of hucMSC-Ex on liver injury development remain elusive. Mouse models of acute and chronic liver injury and liver tumor were induced by carbon tetrachloride (CCl4) injection, followed by administration of hucMSC-Ex via the tail vein. Alleviation of liver injury by hucMSC-Ex was determined. We further explored the production of oxidative stress and apoptosis in the development of liver injury and compared the antioxidant effects of hucMSC-Ex with frequently used hepatic protectant, bifendate (DDB) in liver injury. hucMSC-Ex alleviated CCl4-induced acute liver injury and liver fibrosis and restrained the growth of liver tumors. Decreased oxidative stress and apoptosis were found in hucMSC-Ex-treated mouse models and liver cells. Compared to bifendate (DDB) treatment, hucMSC-Ex presented more distinct antioxidant and hepatoprotective effects. hucMSC-Ex may suppress CCl4-induced liver injury development via antioxidant potentials and could be a more effective antioxidant than DDB in CCl4-induced liver tumor development.

Brain-derived neurotrophic factor preserves intestinal mucosal barrier function and alters gut microbiota in mice.

The intestinal mucosal barrier (IMB) enables the intestine to provide adequate containment of luminal microorganisms and molecules while preserving the ability to absorb nutrients. In this study, we explored the effect of brain-derived neurotrophic factor (BDNF) on IMB function and gut microbiota in mice. BDNF gene knock-out mice (the BDNF+/- group) and wild-type mice (the BDNF+/+ group) were selected. The gut microbiota of these mice was analyzed by denaturing gradient gel electrophoresis (DGGE) assay. The ultrastructure of the ileum and the colonic epithelium obtained from decapitated mice were observed by transmission electron microscopy. The protein expression of epithelial tight junction proteins, zonula occludens-1 (ZO-1) and occludin was detected by immunohistochemistry staining. The protein expression of claudin-1 and claudin-2 was determined by Western blotting. The DGGE band patterns of gut microbiota in the BDNF+/- group were significantly different from that in the BDNF+/+ group, which indicated that the BDNF expression alters the gut microbiota in mice. Compared with the BDNF+/+ group, the BDNF+/- group presented no significant difference in the ultrastructure of ileal epithelium; however, a significant difference was observed in the colonic epithelial barrier, manifested by decreased microvilli, widening intercellular space and bacterial invasion. Compared with the BDNF+/+ group, the expression of ZO-1 and occludin in the BDNF+/- group was significantly decreased. The expression of claudin-1 in the BDNF+/- group was significantly reduced, while the expression of claudin-2 was elevated. These findings indicate that BDNF preserves IMB function and modulates gut microbiota in mice.

Elevated Lipoprotein (a) and Risk of Poor Functional Outcome in Chinese Patients with Ischemic Stroke and Type 2 Diabetes.

The aim of this study was to evaluate the short-term prognostic value of early measurement of serum lipoprotein (a) [Lp(a)] levels in Chinese patients with type 2 diabetes (T2D) and acute ischemic stroke (AIS). The study population comprised 232 consecutive patients with an AIS diagnosis complicated with T2D. Functional outcome was obtained on month 3 according to the modified Rankin Scale (mRS). Unfavorable functional outcome was defined as a mRS score of 3 to 6 points. The prognostic value of Lp(a) at admission to predict the unfavorable functional outcome 3 months after stroke onset was compared with the National Institutes of Health Stroke Scale score and other known outcome predictors. The Lp(a) levels in those patients were obtained with a median value of 16.8 mg/dl (IQR, 9.5-34.4 mg/dl). At 3-month follow-up, an unfavorable functional outcome was found in 86 patients (37.1%). In multivariate models comparing the second (Q2), third, and fourth quartiles against the first quartile of Lp(a), concentrations of Lp(a) in Q2, Q3, and Q4 were associated with unfavorable outcome, and increased risk of unfavorable outcome by 42, 131, and 211%. Interestingly, an elevated Lp(a, > 30 mg/dl) was also associated with unfavorable outcome, and with adjusted OR of 2.25 (95% CI 1.39-3.68). The AUC was significantly increased by adding Lp(a) to established risk factors (difference, 0.041 [95% CI, 0.034-0.053]; P = 0.02). The addition of Lp(a) to established risk factors significantly improved net reclassification improvement and integrated discrimination improvement. Higher Lp(a) levels at admission were associated with increased risk of unfavorable functional outcome and might be useful in identifying stroke patients with T2D at risk for unfavorable functional outcome for early prevention strategies.

Effects of SCN9A gene modification on Na+ channel and the expression of nerve growth factor in a rat model of diarrhea­predominant irritable bowel syndrome.

The aim of the present study was to identify whether the sodium voltage-gated channel alpha subunit 9 (SCN9A) gene modification is a potential treatment for diarrhea­predominant irritable bowel syndrome (D­IBS), via regulating the Na+ channel and the expression of nerve growth factor (NGF). The recombinant adenovirus vector of the SCN9A gene was established, and rat colon cells were isolated for SCN9A gene modification. All subjects were divided into four groups: i) The SCN9A­modified (D­IBS rat model implanted with SCN9A­modified colon cells), ii) negative control (NC; D­IBS rat model implanted with colon cells without SCN9A gene modification), iii) blank (D­IBS rat model without any treatment) and iv) normal (normal rats without any treatment). Western blotting and reverse transcription­quantitative polymerase chain reaction were used to detect the protein and mRNA expression levels of SCN9A, NGF and voltage gated sodium channels (Nav)1.8 and Nav1.9 in rat colon tissues. Compared with the normal group, the rats in the SCN9A, NC and blank groups had significantly elevated mRNA and protein expression levels of NGF, SCN9A, Nav1.8 and Nav1.9. The rats in the SCN9A group demonstrated significantly increased mRNA and protein expression levels of NGF, SCN9A, Nav1.8 and Nav1.9 compared with the NC group and the blank group (all P<0.05). SCN9A gene modification can promote the expression of Nav1.8 and Nav1.9 channels, in addition to NGF which may provide a novel therapeutic basis for treating of D­IBS.

Blockade of angiopoietin-2/Tie2 signaling pathway specifically promotes inflammation-induced angiogenesis in mouse cornea.

AIM: To investigate angiopoietin-2 (Ang-2)/Tie2 signaling pathway involving in inflammatory angiogenesis. METHODS: Three interrupted 11-0 nylon sutures were placed into the corneal stroma of BALB/c mice (6wk old) to induce inflammatory neovascularization. Expression of Ang-2 and Tie2 protein on neovascularization were examined by immunofluorescence. The dynamic expression of Ang-2 mRNA on neovascularization was examined by quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Finally, the mouse model of suture-induced corneal neovascularization was used to assess the role of Ang-2/Tie2 signaling pathway in inflammatory angiogenesis by systemic application of L1-10, an Ang-2 specific inhibitor. Mouse corneal hemangiogenesis were evaluated by whole mount immunofluorescence. RESULTS: Both Ang-2 and Tie2 were expressed on newly generated blood vessels in inflammatory cornea. Ang-2 expression was gradually upregulated around 2wk following injury, which was concurrent with an increased number of blood vessels. Blockade of Ang-2/Tie2 signaling pathway obviously promoted angiogenesis in inflammatory cornea. CONCLUSION: Ang-2/Tie2 signaling pathway seems to play an important role during angiogenesis in inflammatory cornea. This may open new therapeutic applications in pathological processes such as corneal graft survival, wound healing and carcinogenesis.