Exploring the impact of initial moisture content on microbial community and flavor generation in Xiaoqu baijiu fermentation.

Moisture is essential in microbiota succession and flavor formation during baijiu fermentation. However, it remains unknown how moisture content affects microbiota, metabolism, and their relationship. Here, we compared the difference in volatiles, microbiota characteristics, and potential functions with different initial moisture contents (50 %, 55 %, 60 %, 65 %, 70 %). Results showed that the ratio of ethyl acetate to ethyl lactate and total volatile compounds content increased as the moisture content was elevated from 50 % to 70 %. As increasing moisture content, fermentation system microbiota dominated by Lactobacillus was formed more rapidly. Lactobacillus, Dekkera, and Pediococcus were positively correlated with moisture, promoting the production of propanol, acetic acid, butyric acid, and 2-butanol. The complexity and stability of ecological networks enhanced as moisture content increased (R2 = 0.94, P = 0.004). Our study revealed that moisture-drive microbiota was a critical contributor to flavor formation, providing the theoretical basis for moisture control to regulate flavor compounds.

Oenological characteristics of two indigenous Starmerella bacillaris strains isolated from Chinese wine regions.

To broaden knowledge about the oenological characteristics of Starmerella bacillaris, the influence of two Chinese indigenous S. bacillaris strains on the conventional enological parameters and volatile compounds of Cabernet Sauvignon wines were investigated under different inoculation protocols (single inoculation and simultaneous/sequential inoculation with the commercial Saccharomyces cerevisiae EC1118). The results showed that the two S. bacillaris strains could complete alcohol fermentation alone under high sugar concentrations while increasing the content of glycerol and decreasing the content of acetic acid. Compared with wines fermented by EC1118 single inoculation, S. bacillaris single inoculation and S. bacillaris/EC1118 sequential inoculation increased the contents of isobutanol, ethyl isobutanoate, terpenes, and ketones and decreased the contents of isopentanol, phenylethyl alcohol, fatty acids, acetate esters, and total ethyl esters. Furthermore, for S. bacillaris/EC1118 simultaneous inoculation, the concentrations of ethyl esters were increased, contributing to a higher score of "floral" and "fruity" notes in agreement with sensory analysis. KEY POINTS: • S. bacillaris single and simultaneous/sequential inoculation. • Conventional enological parameters and volatile compounds were investigated. • S. bacillaris/EC1118 simultaneous fermentation increased ethyl esters.

Application Potential of Baijiu Non-Saccharomyces Yeast in Winemaking Through Sequential Fermentation With Saccharomyces cerevisiae.

To explore the potential application of non-Saccharomyces yeasts screened from Baijiu fermentation environment in winemaking, the effect of four Baijiu non-Saccharomyces yeasts (two Zygosaccharomyces bailii and two Pichia kudriavzevii) sequentially fermented with Saccharomyces cerevisiae on the physicochemical parameters and volatile compounds of wine was analyzed. The results indicated that there was no obvious antagonism between S. cerevisiae and Z. bailli or P. kudriavzevii in sequential fermentations, and all strains could be detected at the end of alcoholic fermentation. Compare with S. cerevisiae pure fermentation, Z. bailii/S. cerevisiae sequential fermentations significantly reduced higher alcohols, fatty acids, and ethyl esters and increased acetate esters; P. kudriavzevii/S. cerevisiae sequential fermentations reduced the contents of C6 alcohols, total higher alcohols, fatty acids, and ethyl esters and significantly increased the contents of acetate esters (especially ethyl acetate and 3-methylbutyl acetate). Sequential fermentation of Baijiu non-Saccharomyces yeast and S. cerevisiae improved the flavor and quality of wine due to the higher ester content and lower concentration of higher alcohols and fatty acids, non-Saccharomyces yeasts selected from Baijiu fermentation environment have potential applications in winemaking, which could provide a new strategy to improve wine flavor and quality.

Dumbbell structure probe-triggered rolling circle amplification (RCA)-based detection scaffold for sensitive and specific neonatal infection-related small extracellular vesicle (sEV) detection.

Small extracellular vesicles (sEVs) have been reported to play important roles in cell-to-cell communication and are promising biomarkers for the early diagnosis of infections. Therefore, it is in high demand to develop a method that can integrate easy-to-operate sEV isolation and sensitive quantification. We herein propose a novel detection scaffold for sEV isolation via low-speed centrifugation and the quantification of sEVs through DNAzyme-based signal amplification. The detection scaffold is established through dumbbell probe-based RCA (rolling circle amplification), containing repeated CD63 aptamer sections and DNAzyme sections. The original state of the DNAzyme section is locked in a hairpin structure in the detection scaffold. In the presence of sEVs, the CD63 aptamer recognizes and binds with sEVs, leading to the aggregation of sEVs, which can be isolated by low-speed centrifugation and the exposure of the DNAzyme section. After the catalytic fluorescence signal generation from the DNAzyme-based molecular beacon (MB) cleavage, the method exhibited a detection range of 102 to 106 particles per µL. Considering the high sensitivity and wash-free and easy-to-operate features, the strategy reported herein paves a new avenue for the effective determination of sEVs and other membrane biomolecules in fundamental and applied research.

Construction of Dual-Target Recognition-Based Specific MicroRNA Detection Method for Acute Pancreatitis Analysis.

MicroRNAs (miRNAs) play crucial roles in regulating various biological processes and are considered promising biomarkers for clinical diagnosis and therapy of acute pancreatitis. Herein, we present a duplex-specific nuclease (DSN enzyme) and DNAzyme-assisted fluorescent miRNA detections assay that can provide improved detection specificity due to a design of dual-target recognition and a comparable sensitivity. The dual-target recognitions are composed of (i) miRNA unfold hairpin structure toehold to form DNA-RNA duplex, among which the DNA section will be digested by DSN enzyme, releasing miRNA to participant in a next recycle. (ii) After DNAzyme-based nicking site formation in loop section of molecular beacon (MB), miRNA can bind with the loop section of MB and gradually unfold MB probe, generating fluorescence signals. With this general principle, distinct discrimination capability towards even one base pair mismatch of homogenous miRNA is obtained, showing a promising prospect in clinical diagnosis and therapy of acute pancreatitis.

Hepatoprotective mechanism of Silybum marianum on nonalcoholic fatty liver disease based on network pharmacology and experimental verification.

The study aimed to identify the key active components in Silybum marianum (S. marianum) and determine how they protect against nonalcoholic fatty liver disease (NAFLD). TCMSP, DisGeNET, UniProt databases, and Venny 2.1 software were used to identify 11 primary active components, 92 candidate gene targets, and 30 core hepatoprotective gene targets in this investigation, respectively. The PPI network was built using a string database and Cytoscape 3.7.2. The KEGG pathway and GO biological process enrichment, biological annotation, as well as the identified hepatoprotective core gene targets were analyzed using the Metascape database. The effect of silymarin on NAFLD was determined using H&E on pathological alterations in liver tissues. The levels of liver function were assessed using biochemical tests. Western blot experiments were used to observe the proteins that were expressed in the associated signaling pathways on the hepatoprotective effect, which the previous network pharmacology predicted. According to the KEGG enrichment study, there are 35 hepatoprotective signaling pathways. GO enrichment analysis revealed that 61 biological processes related to the hepatoprotective effect of S. marianum were identified, which mainly involved in response to regulation of biological process and immune system process. Silymarin was the major ingredient derived from S. marianum, which exhibited the hepatoprotective effect by reducing the levels of ALT, AST, TC, TG, HDL-C, LDL-C, decreasing protein expressions of IL-6, MAPK1, Caspase 3, p53, VEGFA, increasing protein expression of AKT1. The present study provided new sights and a possible explanation for the molecular mechanisms of S. marianum against NAFLD.

[Analysis of the Risk Factors for Hemorrhagic Cystitis after Hematopoietic Stem Cell Transplantation].

OBJECTIVE: To analyze the risk factors affecting hemorrhagic cystitis(HC) after allogeneic hematopoietic stem cell transplantation(allo-HSCT). METHODS: The clinical data of 153 patients underwent allogeneic hematopoietic stem cell transplantation in the First Affiliated Hospital of Xi'an Jiaotong University from January 2010 to December 2018 were selected and retrospectively analyzed. The incidence, median time and treatment outcome of HC should be observed. Multivariate analysis was used to observe the risk factors of HC in patients, including sex, age, diagnosis, disease status before transplantation, transplantation type, ATG and CTX in the pretreatment scheme, stem cell source, neutrophil and platelet implantation time; CMV, EBV and BKV infection, and acute graft-versus-host disease(aGVHD). RESULTS: Among 153 patients underwent allogeneic hematopoietic stem cell transplantation, 25 (16.34%) patients had HC, the median occurance time was 31 days, all patients achieved complete remission after treatment, no bladder irritation and bladder contracture were left. The results of univariate and multivariate Logistic regression analysis showed that the type of transplantation, ATG, CMV viremia before treatment, aGVHD (r=1.036, 3.234, 3.298 and 2.817, respectively) were the independent risk factors of HC. CONCLUSION: The urinary BKV detections in the patients with HC are positive, mainly occured during the period from day +13 to days +56. HLA haplotype, pretreatment including ATG, and CMV viremia, and aGVHD are the independent risk factors for HC after allo-HSCT.

High proportions of CD3+ T cells in grafts delayed lymphocyte recovery and reduced overall survival in haploidentical peripheral blood stem cell transplantation.

T cells in grafts serve an important role in the pathogenesis of graft versus host disease (GVHD) and immune recovery during HLA matched allogeneic stem cell transplantation. However, the role of T cells in the haploidentical peripheral blood stem cell transplantation (Haplo-PBSCT) is yet to be determined. In the present study, the role of CD3+ T cells in grafts and impact on hematopoietic and immune recovery, cytomegalovirus (CMV) reactivation, GVHD, relapse, progress free survival and overall survival (OS) were evaluated and analyzed. A total of 30 patients who underwent haplo-PBSCT were included in the present study. CD3+ T cells accounted for a median of 23.1% (range 8-47.4%) with a median dose of 299.7x106/kg (range 104-623.4). Patients were divided into two groups according to the CD3+ T cell count: Above the median (high T cell group) and below the median CD3+ T cell (low T cell group). No significant difference was identified between neutrophil and platelet recovery time between two groups (P>0.05). The mean lymphocyte recovery time of high T cell group and low T cell group were 107.07 days (95% CI 79.88-134.25), and 50.4 days (95% CI 41.42-59.38), respectively. The lymphocyte recovery time of high T cell group was higher that of low T cell group (P<0.05). No significant difference between CMV reactivation, chronic GVHD and primary disease relapse rates was observed between two groups (P>0.05). The cumulative incidence of grade II or above acute GVHD was higher in the high T groups compared with low T groups (P<0.05). The overall survival and progress free survival rates were higher in the low T cell group compared with the high T cell group (P<0.05). In conclusion, high levels of CD3+ T cells in the grafts were associated with delayed lymphocyte recovery and an increased risk of acute GVHD and decreased overall survival.

[Expression of DNMT3b Gene in Myeloma RPMI 8226 Cells and Its Biological Significance].

OBJECTIVE: To study the expression of DNMT3b gene in myeloma RPMI8226 cells and its biological significance. METHODS: The activity of DNA methyltransferase was detected by ELISA, and the expression of DNMT3b in RPMI8226 cells was analyzed by semi-quantitative RT-PCR and real-time fluorescent quantitative PCR. The proliferation and expression of DNMT3b gene in RPMI8226 cells intervened with capecitabine for 24 hours were detected. RESULTS: The activity of DNMT and expression of DNMT3b in RPMI 8226 cells increased. The proliferation of RPMI8226 cells was inhibited, and the apoptosis occurred in RPMI 8226 cells intervened with capecitabine for 24 hours. The expression level of DNMT3b gene was decreased after being intervened with capecitabine for 24 hours. CONCLUSION: The expression level of DNMT3b in myeloma RPMI 8226 cells increase, and capecitabine can inhibit the proliferation of RPMI 8226 and induce apoptosis by inhibiting the expression of DNMT3b gene. Therefore, DNMT3b is expected to be a new target for myeloma therapy.

[Risk Factors and Therapeutic Outcome of Intestinal Acute Graft Versus Host Disease in Acute Leukemia Patients after Haploidentical Peripheral Hematopoietic Stem Cell Transplantation].

OBJECTIVE: To investigate the risk factors and therapeutic outcome of acute graft versus host disease (aGVHD) in patients with acute leukemia after haploidentical peripheral hematopoietic stem cell transplantation. METHODS: The clinical data of 19 cases of acute leukemia underwent haploidentical hematopoietic stem cell transplanttion during January 2010 and December 2010 were retrospectively analyzed. The effects of patients sex, donor-recipient sex difference, donor age, conditioning regimen, dosage of anti-thymocyte globulin(ATG), mononuclear cell and CD34+ cell counts on the intestinal aGVHD were analyzed by Logistic regression. RESULTS: Intestinal aGVHD occurred in 5 cases with 1 case at stage II 3 cases at stage III and 1 case at stage IV on the 7th, 22th, 27th, 70th and 154th day after transplantation, respectively. Single factor analysis showed that the patient's sex, donor-recipient sex difference, donor age, dosage of ATG, mononuclear cell and CD34+ cell counts were not related with the occurrence of the intestinal aGVHD, and the conditoning regimen was the risk factor for the intestinal aGVHD. 2 cases among 5 cases with intestinal aGVHD were treated with methylprednisolone at dosage of 1 mg/kg per day, 1 case was treated with methylprednisolone therapy combined with tacrolimus. 2 cases of methylprednisolone-resistance were treated with CD25 monoclonal antibody. Intestinal aGVHD of all patients was improved after the above-mentioned treatment. CONCLUSION: Conditioning regimen of haploidentical peipheral hematopoieitc stem cell transplantaion has effects on the intestinal aGVHD, which needs to be confirmed by further research.

Therapeutic experience of vincristine/cyclophosphamide/melphalan or mitoxantrone/prednisone combination therapy plus thalidomide as first-line induction therapy for newly diagnosed multiple myeloma in a single institution of China.

INTRODUCTION: To investigate the safety and efficacy of the combination regimen vincristine, cyclophosphamide, melphalan or mitoxantrone and prednisone (VCMP) plus thalidomide as first-line induction therapy for newly diagnosed multiple myeloma (MM). METHODS: Three hundred and ninety-six symptomatic, newly diagnosed MM patients were treated with VCMP plus thalidomide in our hospital for the past 11 years, and clinical data of these patients were retrospectively analyzed. RESULTS: Of the 396 patients enrolled, the total response rate was 77.3%. Forty-three patients relapsed after sCR and CR. Mean cycles to first response were six cycles (range 1-16 cycles). A total of 53% of patients achieved at least a PR within the first cycle of therapy. The actuarial 1-year, 3-year and 5-year overall survival of all patients were 89.4%, 29.5% and 10.6%, respectively. The probabilities of 1-year, 3-year and 5-year progression-free survival of all patients were 84.0%, 23.1% and 8.4%, respectively. The major adverse events were gastrointestinal symptoms, electrolytes and glucose metabolism disorders, hypertension, infection, peripheral nerve disease and hematological adverse events, which were mostly below grade 3 and could be alleviated by symptomatic treatment. CONCLUSION: We concluded that VCMP plus thalidomide is an effective regimen with manageable side effects in the treatment of symptomatic, newly diagnosed MM including elderly patients and patients with renal failure.

[Effect of Different Hemapheresis Procedures on Components of Hematopoietic Stem Cells Collected from Healthy Donors].

OBJECTIVE: To investigate the effects of different hemapheresis procedures on the components of hematopoietic stem cells(HSCs) collected from helathy donors. METHODS: twelve donors were underwent stem cell collection from January 2015 to August 2016, and the stem cells were randomly colleted by AutoPBSC procedure of COBE spectra and MNC procedure of the Spectra Optia blood cell separator, the mononuclear cells, CD34+ cells, granulocytes, lymphocytes and platelets in the collections were compared. RESULTS: The circulating blood volume, the acquisition time and dosage of anticoagulants were not significantly different between two procedures. The volume and the mononuclear cell count collected by AutoPBSC procedure were lower than those by the MNC procedure, while the CD34+ cell count by AutoPBSC procedure was higher than that by the MNC procedure. More lymphocytes and platelets were collected by AutoPBSC procedure as compared with that by the MNC procedure (P<0.05). CONCLUSION: Compared with MNC procedure of the Spectra Optia blood cell separator, the number of collected stem cells, lymphocytes and platelets are higher by using AutoPBSC procedure of the COBE spectra blood cell separator.

[Effects of Hepatovirus B Infection on the Hematopoietic Stem Cell Collection and Implantment in Lymphoma Patients Received Autologous Peripheral Blood Hematopoietic Stem Cell Transplantation].

OBJECTIVE: To investigate the effect of hepatovirus B(HBV) infection on the hematopoietic stem cell collection and implantment in lymphoma patients received autologous peripheral hematopoietic blood stem cells transplantation. METHODS: Clinical data of 40 lymphoma patients who received autologous peripheral hematopoietic blood stem cell transplantation between January 2006 and October 2014 was analyzed retrospectively. Among 40 patients with lymphoma 8 patients combined with HBV infection were prophylacticly given nucleoside analogues and 32 patients without HBV infection. The counts of mononuclear cells(MNC) and CD34 positive cells were collected and the hematopoietic reconstitution as well as overall survival rates and progress-free survival rates were detected and counted between patients with or without HBV infection. RESULTS: The counts of MNC and CD34 positive cells in all patients were standard, and there was no significant difference between patients with or without HBV infection. HBV wasn't reactivated among the 8 patients with HBV infection. The 1, 3 and 5 years' overall survival rates and progress-free survival rates of patients with HBV infection were 100%, 85.7%, 57.1% and 100%, 80%, 53%, respectively and the 1,3 and 5 years' overall survival rates and progress-free survival rates of patients without HBV infection were 100%, 88.9%, 82.1% and 90%, 90%, 90%, respectively. CONCLUSION: HBV infection may have no effect on the collection of stem cells and hematopoietic reconstitution. Prophylactic use of nucleoside analogues can effectively prevent the hepatitis B virus reactivation, moreover had no effect on the collection and hematopoietic reconstitution.

[Effect of Different Procedures from Blood Cell Separators on Autologous PBSC Collection Efficiency].

OBJECTIVE: To observe the efficacy and adverse reactions of autologous PBSC collection when the autoPBSC procedure and MNC procedure of COBE Spectra cell separator and the MNC procedure of Spectra Optia cell separator were used. METHODS: The autologous perepheral blood hematopoietic stem cells from 41 patients were collected by using autoPBSC procedure and MNC procedure of COBE Spectra blood cell separator and MNC procedure of Spectra Optia blood cell separator. The numbers of MNC and CD34+ cells collected by 3 collected procedure, the difference of hemoglobin (Hb) drop and platelet decrease, and the adverse reaction of patients were observed. RESULTS: When the whole blood processing and the collection time were basically same among these 3 groups, the MNC counts collected by MNC procedure of COBE Spectra and Spectra Optia were higher than that of AutoPBSC procedure of COBE Spctra, but the CD34+ cell count was lower than that collected by AutoPBSC procedure (P< 0.05). The final product volume collected by MNC procedure of COBE Spectra and Spectra Optia was bigger than that collected by AutoPBSC procedure. In comprission with MNC procedure of COBE Spectra cell seperator, the CD34+ count collected by MNC procedure of Spectra Optia Seperator did not show significant difference, but the CD34+ cell count collected by MNC procedure of Spectra Optia was higher than that collected by MNC procedure of COBE Spectra cell separator (P<0.05). The platelet count and hemoglobin level collected by MNC procedure of Spectra Optia were lower than those before collection. The adverse reactions in the 3 procedures were similar, and the patients could tolerate them. CONCLUSION: The AutoPBSC procedure of COBE Spectra and MNC procedure of Spectra Optia are better than MNC procedure of COBE Spectra for autologous peripheral blood hematopoietic stem cells collection. The loss of blood platelet and hemoglobin after collection is lowest in MNC procedure of Spectra Optia.

Dendritic cell-activated cytokine-induced killer cell-mediated immunotherapy is safe and effective for cancer patients >65 years old.

Individuals >65 years old account for a large proportion of cancer patients, and usually have poor prognoses due to relative weaker physiological function and lower drug tolerance. To characterize the efficacy and safety of dendritic cell (DC)-activated cytokine-induced killer cell (CIK)-mediated treatment, and develop an adoptive immunotherapy for cancer patients >65 years old, a retrospective study was performed in 58 cancer sufferers who received 1-4 cycles of DC-activated CIK (DC-CIK) treatment and evaluated the response (tumor remission rate) and toxicity (side effects to the treatment). The present results showed that DCs and CIKs could be expanded rapidly in vitro, and following co-culture with DCs, the population of cluster of differentiation (CD) 3+, CD3+CD4+, CD3+CD8+ and CD3+CD56+ CIKs was significantly increased compared to CIKs without DC activation (P=0.044). In addition, DC-CIK infusion produced marked clinical outcomes, resulting in an objective remission rate, overall clinical benefit rate and Karnofsky performance status of 44.83, 75.86 and 87.28±5.46%, respectively, which was significantly improved compared with prior to treatment (P<0.05). Additionally, subsequent to two cycles of this immunotherapy, several tumor marker expression levels declined, returning to the normal range. The proportion of CD3+CD4+ (P=0.017) and CD3+CD8+ (P=0.023) lymphocytes, and the population of CD4/CD8 cells (P=0.024) were also increased. In conclusion, the present study suggests that the immunotherapy mediated by DC-CIK is safe and effective for cancer patients aged >65 years.

[Relation of reinfused hematopoietic stem cell volume and recipient's leukocyte count at reinfusion with prognosis of disease in allo-hematopoietic stem cell transplantation].

This study was aimed to investigate the relation of reinfused hematopoietic stem cell volume and recipient's leukocyte count at reinfusion with prognosis of disease in allo-hematopoietic stem cell transplantation (allo-HSCT). The clinical data of 37 patients received allo-HSCT in our hospital were analyzed retrospectively. The 37 patients were divided into agranulocytosis and non-agranulocytosis groups according to the recipient's leukocyte count at reinfusion, and were divided into the high dose and low dose groups according to the median number of reinfused mononuclear cells (MNC) and CD34(+) cells. Then, hematopoietic reconstructions,GVHD, relapse and death rates of patients were compared. The results showed that the hematopoietic reconstruction of patients in non-agranulocytosis group and high dose MNC group were earlier than that in agranulocytosis group and low dose MNC group. There was no significant difference of hematopoietic reconstruction between the groups of high dose CD34(+) cells and low dose CD34(+) cells. The GVHD incidence was higher in high dose MNC group and non-agranulocytosis group than that in low dose MNC group and agranulocytosis group (P < 0.05). There were no statistical differences of relapsed and death rates between different reinfused number of HSC and recipient's leukocyte count at reinfusion.It is concluded that the infused MNC number and the recipient's leukocyte count at reinfusion in allo-HSCT correlated with hematopoietic reconstruction, the GVHD incidence is high in high dose MNC and non-agranulocytosis groups, the reinfused HSC volume and the recipient's leukocyte count at reinfusion not significantly relate with relapse and death rates.

Overexpression of hypoxia-inducible factor-1α exacerbates endothelial barrier dysfunction induced by hypoxia.

BACKGROUND/AIMS: The mechanisms involved in endothelial barrier dysfunction induced by hypoxia are incompletely understood. There is debate about the role of hypoxia-inducible factor-1α (HIF-1α) in endothelial barrier disruption. The aim of this study was to investigate the effect of genetic overexpression of HIF-1α on barrier function and the underlying mechanisms in hypoxic endothelial cells. METHODS: The plasmid pcDNA3.1/V5-His-HIF-1α was stably transfected into human endothelial cells. The cells were exposed to normoxia or hypoxia. The mRNA and protein expressions of HIF-1α were detected by RT-PCR and Western blot respectively. The barrier function was assessed by measuring the transendothelial electrical resistance (TER). The Western blot analysis was used to determine the protein expression of glucose transporter-1 (GLUT-1), zonular occludens-1 (ZO-1), occludin, and myosin light chain kinase (MLCK) in endothelial cells. The mRNA expression of proinflammatory cytokines was detected by qRT-PCR. RESULTS: Genetic overexpression of HIF-1α significantly increased the mRNA and protein expression of HIF-1α in endothelial cells. The overexpression of HIF-1α enhanced the hypoxia-induced increase of HIF-1α and GLUT-1 protein expression. HIF-1α overexpression not only exacerbated hypoxia-induced endothelial barrier dysfunction but also augmented hypoxia-induced up-regulation of MLCK protein expression. HIF-1α overexpression also enhanced IL-1ß, IL-6 and TNF-α mRNA expression. CONCLUSION: We provide evidence that genetic overexpression of HIF-1α aggravates the hypoxia-induced endothelial barrier dysfunction via enhancing the up-regulation of MLCK protein expression caused by hypoxia, suggesting a potential role for HIF-1α in the pathogenesis of endothelial barrier dysfunction in hypoxia.

Amelioration of IFN-γ and TNF-α-induced intestinal epithelial barrier dysfunction by berberine via suppression of MLCK-MLC phosphorylation signaling pathway.

Intestinal barrier dysfunction occurs in many intestinal diseases, in which proinflammatory cytokines play critical roles. However, researchers are still on the way to defining the underlying mechanisms and to evaluate therapeutic strategies for restoring intestinal barrier function. Berberine, a drug that has clinically been used to treat gastroenteritis and diarrhea for thousands of years, has been shown to protect barrier function in both endothelial and epithelial cells, but the mechanisms are completely unknown. In this study, we investigate the protective actions of berberine on barrier function and the underlying mechanisms in Caco-2 monolayers challenged with IFN-γ and TNF-α. Caco-2 monolayers were treated without or with simultaneous IFN-γ and TNF-α in the absence or presence of berberine. Both transepithelial electrical resistance (TER) and paracellular permeability were measured to evaluate barrier function. The expression and distribution of tight junction proteins ZO-1, occluding, and claudin-1 were respectively analyzed by immunoblot or immunofluorescence. The expressions of phosphorylated myosin light chain (pMLC), MLC kinase (MLCK) and hypoxia-inducible factor-1α (HIF-1α) were determined by immunoblot. The translocation of NF-κB p65 to nuclei was analyzed by immunofluorescence and immunoblot, respectively. The results showed that berberine significantly attenuated TER decrease and paracellular permeability increase in Caco-2 monolayers treated with IFN-γ and TNF-α. Berberine also dramatically alleviated IFN-γ and TNF-α-induced morphological alteration of tight junction proteins ZO-1, occluding, and claudin-1. The increase of both MLC phosphorylation and MLCK protein expression induced by IFN-γ and TNF-α was significantly inhibited by berberine treatment. Additionally, berberine suppressed the activation of HIF-1α, but not NF-κB. Taken together, it is suggested that berberine attenuates IFN-γ and TNF-α-induced intestinal epithelial barrier dysfunction by inhibiting the signaling pathway of MLCK-dependent MLC phosphorylation mediated by HIF-1α.

A clinical analysis of erythrocytapheresis for the treatment of polycythemia.

PURPOSE: To evaluate the efficacy and safety of erythrocytapheresis (ECP) in the treatment of polycythemia. METHODS: Patients diagnosed with polycythemia were included in this retrospective analysis and treated with ECP (n=20) or conventional treatments (exsanguination; n=20). Blood laboratory values and adverse effects were recorded. RESULTS: In ECP-treated patients mean red blood cell (RBC) collection time was 25.7±4.5min (range: 19-37min), with a mean collection volume of 773.5±129.3mL (range: 600-1002mL). From baseline, ECP reduced the mean number of RBCs (0.6×10(12)/L [7.6%]), mean hemoglobin (31.1g/L [14.8%]), and mean hematocrit (13.1% [20.2%]) (P<0.001 for each). After ECP, a marked reduction in symptoms associated with polycythemia was also observed. CONCLUSIONS: Treatment of patients with polycythemia using ECP reduces RBC count, hemoglobin, and hematocrit. The advantages associated with ECP over conventional therapy should be considered when choosing a treatment plan for patients with polycythemia.