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This study aimed to investigate the effects of probiotic Lactobacillus paracasei NL41 on inflammation and the gut microbiota of type 2 diabetic (T2D) rats induced by high-fat diet (HFD) and low-dose streptozotocin (STZ). A T2D rat model was established by inducing Sprague-Dawley rats with HFD/STZ, followed by 12-weeks L. paracasei NL41 gavage. The blood, colonic tissues, and feces samples of these rats were collected for inflammation, histology, and intestinal microbiota profiling. L. paracasei NL41 treatment induced remarkable improvement in the inflammatory status by decreasing the levels of serum lipopolysaccharides (LPS), free fatty acids (FFA), tumor necrosis factor-α (TNF-α), interleukin (IL)-6, and IL-8 and increasing the level of IL-10. Gut barrier function was significantly protected in NL41-treated rats. Moreover, the strain NL41 induced changes in the microbiota structure and influenced the relative abundance of the key species. Specifically, Bacteroides, Clostridia (specifically, Ruminococcus torques), and Parasutterella were significantly reduced, while some beneficial microorganisms (Bacteroidales_S24-7_group and the families Lachnospiraceae and Ruminococcaceae) were enriched by NL41. The correlational analyses indicated that L. paracasei NL41 ameliorating inflammation was closely related to the key species of the gut microbiota. The present study indicates that probiotic L. paracasei NL41 decreases LPS-induced inflammation by improving the gut microbiota and preserving intestinal integrity.

ObjectivePatients with renal failure suffer from symptoms caused by uraemic toxins, possibly of gut microbial origin, as deduced from studies in animals. The aim of the study is to characterise relationships between the intestinal microbiome composition, uraemic toxins and renal failure symptoms in human end-stage renal disease (ESRD).DesignCharacterisation of gut microbiome, serum and faecal metabolome and human phenotypes in a cohort of 223 patients with ESRD and 69 healthy controls. Multidimensional data integration to reveal links between these datasets and the use of chronic kidney disease (CKD) rodent models to test the effects of intestinal microbiome on toxin accumulation and disease severity.ResultsA group of microbial species enriched in ESRD correlates tightly to patient clinical variables and encode functions involved in toxin and secondary bile acids synthesis; the relative abundance of the microbial functions correlates with the serum or faecal concentrations of these metabolites. Microbiota from patients transplanted to renal injured germ-free mice or antibiotic-treated rats induce higher production of serum uraemic toxins and aggravated renal fibrosis and oxidative stress more than microbiota from controls. Two of the species, Eggerthella lenta and Fusobacterium nucleatum, increase uraemic toxins production and promote renal disease development in a CKD rat model. A probiotic Bifidobacterium animalis decreases abundance of these species, reduces levels of toxins and the severity of the disease in rats.ConclusionAberrant gut microbiota in patients with ESRD sculpts a detrimental metabolome aggravating clinical outcomes, suggesting that the gut microbiota will be a promising target for diminishing uraemic toxicity in those patients.Trial registration numberThis study was registered at ClinicalTrials.gov (NCT03010696).

N6 -methyladenosine (m6 A) is a chemical modification present in multiple RNA species and is most abundant in mRNAs. Studies on m6 A reveal its comprehensive roles in almost every aspect of mRNA metabolism, as well as in a variety of physiological processes. Although some recent discoveries indicate that m6 A can affect the life cycles of numerous viruses as well as the cellular antiviral immune response, the roles of m6 A modification in type I interferon (IFN-I) signaling are still largely unknown. Here, we reveal that WT1-associated protein (WTAP), one of the m6 A "writers", is degraded via the ubiquitination-proteasome pathway upon activation of IFN-I signaling. With the degradation of WTAP, the m6 A levels of IFN-regulatory factor 3 (IRF3) and interferon alpha/beta receptor subunit 1 (IFNAR1) mRNAs are reduced, leading to translational suppression of IRF3 and instability of IFNAR1 mRNA. Thus, the WTAP-IRF3/IFNAR1 axis may serve as negative feedback pathway to fine-tune the activation of IFN-I signaling, which highlights the roles of m6 A in the antiviral response by dictating the fate of mRNAs associated with IFN-I signaling.

Cervical cancer is the fourth most common malignant tumor in women worldwide. The persistent infection of high-risk Human Papillomavirus (hrHPV) is considered to be the primary cause of this disease. As an innate immune receptor, the nucleotide-binding oligomerization domain protein-1 (NOD1) recognizes the pathogen-associated molecular pattern (PAMP), subsequently initiating immune responses. NOD1 is also involved in the apoptotic signaling pathway and mutates in many cancer cells. In the study, we revealed that NOD1 expression decreased during the progression of cervical intraepithelial neoplasia to cervical cancer and that HPV16 E6/E7 oncoproteins induced down-regulation of NOD1. Moreover, the activation of NOD1 promoted the apoptosis of HPV16-positive cervical cancer cells. The data indicated that the dysregulation of NOD1-mediated inflammation and apoptosis may contribute to cervical intraepithelial neoplasia progression and cervical cancer.

In the past two decades, emerging studies have suggested that DExD/H box helicases belonging to helicase superfamily 2 (SF2) play essential roles in antiviral innate immunity. However, the antiviral functions of helicase SF1, which shares a conserved helicase core with SF2, are little understood. Here we demonstrate that zinc finger NFX1-type containing 1 (ZNFX1), a helicase SF1, is an interferon (IFN)-stimulated, mitochondrial-localised dsRNA sensor that specifically restricts the replication of RNA viruses. Upon virus infection, ZNFX1 immediately recognizes viral RNA through its Armadillo-type fold and P-loop domain and then interacts with mitochondrial antiviral signalling protein to initiate the type I IFN response without depending on retinoic acid-inducible gene I-like receptors (RLRs). In short, as is the case with interferon-stimulated genes (ISGs) alone, ZNFX1 can induce IFN and ISG expression at an early stage of RNA virus infection to form a positively regulated loop of the well-known RLR signalling. This provides another layer of understanding of the complexity of antiviral immunity.

Background & Aims Human induced pluripotent stem cell (hiPSC)-derived liver modeling systems have the potential to overcome the shortage of donors for clinical application and become a model for drug development. Although several strategies are available to generate hepatic micro-tissues, few have succeeded in generating a liver organoid with hepatobiliary structure from hiPSCs. Methods At differentiation stages I and II (day 1–15), 25% of mTeSR™ culture medium was added to hepatic differentiation medium to induce endodermal and mesodermal commitment and thereafter hepatic and biliary co-differentiation. At stage III (day 15–45), 10% cholesterol+ MIX was added to the maturation medium to promote the formation and maturation of the hepatobiliary organoids. Phenotypes and functions of organoids were determined by specific markers and multiple functional assays both in vitro and in vivo. Results In this system, hiPSCs were induced to form 3D hepatobiliary organoids and to some extent recapitulated key aspects of early hepatogenesis in a parallel fashion. The organoids displayed a series of functional attributes. Specifically, the induced hepatocyte-like cells could take up indocyanine green, accumulate lipid and glycogen, and displayed appropriate secretion ability (albumin and urea) and drug metabolic ability (CYP3A4 activity and inducibility); the biliary structures in the system showed gamma glutamyltransferase activity and the ability to efflux rhodamine and store bile acids. Furthermore, after transplantation into the immune-deficient mice, the organoids survived for more than 8 weeks. Conclusion This is the first time that functional hepatobiliary organoids have been generated from hiPSCs. The organoid model will be useful for in vitro studies of the molecular mechanisms of liver development and has important potential in the therapy of liver diseases. Lay summary Herein, we established a system to generate human induced pluripotent stem cell-derived functional hepatobiliary organoids in vitro, without any exogenous cells or genetic manipulation. To some extent this model was able to recapitulate several key aspects of hepatobiliary organogenesis in a parallel fashion, holding great promise for drug development and liver transplantation.

The conventional planar culture of adherent cells is inefficient for large‐scale manufacturing of cell and gene therapy products. We developed a facile and efficient bead‐to‐bead cell‐transfer method for serial subculture and large‐scale expansion of human mesenchymal stem cells (hMSCs) with microcarriers in bioreactors. We first compared culture medium with and without nucleosides and found the former maintained the expression of surface markers of hMSCs during their prolonged culture and enabled faster cell proliferation. Subsequently, we developed our bead‐to‐bead cell transfer method to subculture hMSCs and found that intermittent agitation after adding fresh microcarriers to cell‐populated microcarriers could promote spontaneous cell migration to fresh microcarriers, reduce microcarrier aggregation, and improve cell yield. This method enabled serial subculture of hMSCs in spinner flasks from passage 4 to passage 9 without using proteolytic enzymes, which showed faster cell proliferation than the serial planar cultures undergoing multiple enzyme treatment. Finally, we used the medium containing nucleosides and our bead‐to‐bead cell transfer method for cell culture scale‐up from 4‐ to 50‐L cultures in single‐use bioreactors. We achieved a 242‐fold increase in the number of cells to 1.45 × 1010 after 27‐day culture and found that the cells harvested from the bioreactors maintained proliferation ability, expression of their surface markers, tri‐lineage differentiation potential and immunomodulatory property. This study shows the promotive effect of nucleosides on hMSC expansion and the potential of using our bead‐to‐bead transfer method for larger‐scale manufacturing of hMSCs for cell therapy., The expansion of human mesenchymal stem cells (hMSCs) was promoted using a culture medium with nucleosides. Intermittent agitation was performed to facilitate bead‐to‐bead cell transfer to expand hMSCs in bioreactors without proteolytic enzymes. By using this culture medium and our bead‐to‐bead cell‐transfer method, hMSC cultures were scaled up from 4‐ to 50‐L in bioreactors, resulting in a 242‐fold increase in cell number to 1.45 × 1010.

Background: Most tumors have an enhanced glycolysis flux, even when oxygen is available, called the aerobic glycolysis or the Warburg effect. Metabolic reprogramming promotes cancer progression, and is even related to the tumorigenesis. However, it is not clear whether the observed metabolic changes act as a driver or a bystander in cancer development. Methods: In this study, the metabolic characteristics of oral precancerous cells and cervical precancerous lesions were analyzed by metabolomics, and the expression of glycolytic enzymes in cervical precancerous lesions was evaluated by RT-PCR and Western blot analysis. Results: In total, 115 and 23 metabolites with reliable signals were identified in oral cells and cervical tissues, respectively. Based on the metabolome, oral precancerous cell DOK could be clearly separated from normal human oral epithelial cells (HOEC) and oral cancer cells. Four critical differential metabolites (pyruvate, glutamine, methionine and lysine) were identified between DOK and HOEC. Metabolic profiles could clearly distinguish cervical precancerous lesions from normal cervical epithelium and cervical cancer. Compared with normal cervical epithelium, the glucose consumption and lactate production increased in cervical precancerous lesions. The expression of glycolytic enzymes LDHA, HK II and PKM2 showed an increased tendency in cervical precancerous lesions compared with normal cervical epithelium. Conclusions: Our findings suggest that cell metabolism may be reprogrammed at the early stage of tumorigenesis, implying the contribution of metabolic reprogramming to the development of tumor.

Cytology-based Papanicolaou test on and primary HPV screening have been widely used in the identification of cervical cancer and precancerous lesions, which is of great significance for the prevention and treatment of cervical cancer. Patients diagnosed as ASCUS/LSIL usually need follow-up because some of them may develop into CIN2+. The consequences of women positive for HPV vary from person to person; some of them may progress into cervical dysplasia, reversible forms of precancerous lesions, and eventually invasive cervical cancer. Therefore, it is necessary to establish an effective biomarker to triage different patients according to the preliminary screening results. p16 acts as a cell cycle regulatory protein that induces cell cycle arrest, and Ki-67 is a cell proliferation marker. Under physiological conditions, they could not co-express in the same cervical epithelial cells. The co-expression of these two molecules suggests a deregulation of the cell cycle mediated by HR-HPV infection and predicts the presence of high-grade cervical epithelial lesions. There is increasing evidence that p16/Ki-67 dual-staining cytology can be used as an alternative biomarker, showing overall high sensitivity and specificity for identifying high-grade CIN and cervical cancer. In this review, we discuss the significance of p16/Ki-67 dual-staining and summarize its application in the screening and triaging of cervical cancer and precancerous lesions.

Bifidobacterium longum strain BBMN68 is sensitive to low concentrations of oxygen. A transcriptomic study was performed to identify candidate genes for B. longum BBMN68’s response to oxygen treatment (3%, v/v). Expression of genes and pathways of B. longum BBMN68 involved in nucleotide metabolism, amino acid transport, protein turnover and chaperones increased, and that of carbohydrate metabolism, translation and biogenesis decreased to adapt to the oxidative stress. Notably, expression of two classes of ribonucleotide reductase (RNR), which are important for deoxyribonucleotide biosynthesis, was rapidly and persistently induced. First, the class Ib RNR NrdHIEF was immediately upregulated after 5 min oxygen exposure, followed by the class III RNR NrdDG, which was upregulated after 20 min of exposure. The upregulated expression of branched-chain amino acids and tetrahydrofolate biosynthesis-related genes occurred in bifidobacteria in response to oxidative stress. These change toward to compensate for DNA and protein damaged by reactive oxygen species (ROS). In addition, oxidative stress resulted in improved B. longum BBMN68 cell hydrophobicity and autoaggregation. These results provide a rich resource for our understanding of the response mechanisms to oxidative stress in bifidobacteria.

Reconstruction of large bone defects remains a clinical challenge because current approaches involving surgery and bone grafting often do not yield satisfactory outcomes. For artificial bone substitutes, angiogenesis plays a pivotal role to achieve the final success of newly regenerated bone. In this study, dexamethasone-loaded biphasic calcium phosphate nanoparticles/collagen composite scaffolds with several types of concave microgrooves were prepared for simultaneous promotion of angiogenesis and osteogenesis. Microgrooves in the scaffolds were supposed to guide the assembly of human umbilical vascular endothelial cells (HUVECs) into well aligned tubular structures, thus promoting rapid angiogenesis. The scaffolds were used for co-culture of HUVECs and human bone marrow-derived mesenchymal stem cells. Subcutaneous implantation in mice showed that more blood vessels and newly formed bone were observed in the microgrooved composite scaffolds than in the control scaffold. Scaffold bearing parallel microgrooves with a concave width of 290 µm and a convex ridge width of 352 µm showed the highest promotion effect on angiogenesis and osteogenesis among the parallelly microgrooved composite scaffolds. The scaffolds bearing a grid network had further superior promotion effect to the scaffolds bearing parallel microgrooves. The results indicated that microgrooves in the composite scaffolds facilitated angiogenesis and stimulated new bone formation. The microgrooved composite scaffolds should be useful for repairing of large bone defects.

Dipeptidyl peptidase-IV (DPP-IV) is a serine exo-peptidase that can inactivate incretins by removing N-terminal dipeptides. Currently, inhibiting the DPP-IV activity is a common treatment for type 2 diabetes (T2D). The goal of this study is to investigate whether IADHFL, a novel DPP-IV inhibitory peptide identified from bighead carp (Hypophthalmichthys nobilis), has the potential to modulate T2D. IADHFL remained stable after simulated gastrointestinal digestion and significantly decreased the activity and expression of both soluble and membrane-bound DPP-IV after 24 h and 48 h of treatment. Intact peptide absorption was observed, but a percentage of the peptide was degraded while passing through a monolayer of Caco-2 cells. In addition, a double-layered cell model showed that the peptide could increase insulin secretion from INS-1 cells after glucose treatments of 2.8 mM and 16.7 mM. Finally, IADHFL could regulate the expression levels of genes associated with insulin secretion and T2D in INS-1 cells.

Enhanced glycolysis under normoxic conditions is known as aerobic glycolysis or the Warburg effect and is a hallmark of many tumors. Viral infection may also induce aerobic glycolysis as it is required for replication and survival. Tumor viruses inducing aerobic glycolysis and lactate production during latent infection suggest a potential role of virus-induced glycolysis in tumorigenesis. Virus or virus-encoded proteins regulate glucose uptake and lactate export, increase the activity of glycolytic enzymes, and modulate glucose metabolic signals. Accumulating evidence suggests that virus-induced glycolysis may facilitate cell growth, transformation, migration, and invasion, but its significance in tumorigenesis remains unclear. We summarize the effects of oncogenic viruses on the metabolic shift to aerobic glycolysis and discuss the possible association of this metabolic reprogramming with tumor development and progression.

T cells are activated and differentiated into Th cells depending on the rapid and accurate changes in the cell transcriptome. In addition to changes in mRNA expression, the sequences of many transcripts are altered by alternative splicing and alternative polyadenylation (APA). We profiled the APA sites of human CD4+ T cell subsets with high-throughput sequencing and found that Th1 cells harbored more genes with shorter tandem 3′ untranslated regions (UTRs) than did naive T cells. We observed that STAT5B, a key regulator of Th1 differentiation, possessed three major APA sites and preferred shorter 3′ UTRs in Th1 cells. In addition, small nuclear ribonucleoprotein polypeptide A (SNRPA) was found to bind directly to STAT5B 3′ UTR and facilitate its APA switching. We also found that p65 activation triggered by TCR signaling could promote SNRPA transcription and 3′ UTR shortening of STAT5B. Thus we propose that the APA switching of STAT5B induced by TCR activation is mediated by SNRPA.

Reprogramming of cellular metabolism is a hallmark of cancers. Cancer cells more readily use glycolysis, an inefficient metabolic pathway for energy metabolism, even when sufficient oxygen is available. This reliance on aerobic glycolysis is called the Warburg effect, and promotes tumorigenesis and malignancy progression. The mechanisms of the glycolytic shift in tumors are not fully understood. Growing evidence demonstrates that many signal molecules, including oncogenes and tumor suppressors, are involved in the process, but how oncogenic signals attenuate mitochondrial function and promote the switch to glycolysis remains unclear. Here, we summarize the current information on several main mediators and discuss their possible mechanisms for triggering the Warburg effect.

Objectives To evaluate the diagnostic values of urine liquid-based cytology (LBC), fluorescence in situ hybridization (FISH), and p16/Ki-67 dual immunostaining in the detection of upper tract urothelial carcinomas (UTUCs). Methods Sixty-one patients with UTUCs were retrospectively analyzed. All patients were diagnosed both by urine cytology and by FISH, and histologically confirmed as UTUCs. Thirty-two patients had been stained with p16/Ki-67 dual labeling. Results The sensitivities for low-grade UTUCs (LGUTUCs) and high-grade UTUCs (HGUTUCs) were 33.3% and 67.4% by LBC, 60% and 69.6% by FISH, and 12.5% and 66.7% by p16/Ki-67 dual labeling, respectively. Conclusions Our results indicated that LBC was more suitable to identify HGUTUCs, FISH was highly valuable for predicting LGUTUCs, and p16/Ki-67 dual labeling was useful for distinguishing HGUTUCs from LGUTUCs. The combined application of these methods may improve the sensitivity or accuracy in the detection or diagnosis of UTUCs.

We investigated bioactive peptides obtained from muscle protein hydrolysate of bighead carp by evaluating in vitro dipeptidyl peptidase IV (DPP-IV) and angiotensin-converting enzyme (ACE) inhibitory capacities and antioxidant activity. Peptide sequences were identified from pepsin hydrolysates. Met-Lys-Ala-Val-Cys-Phe-Ser-Leu was one of the most effective sequences with 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (EC 50 = 4.58 ± 0.15 μM) and relatively high ACE inhibitory capacity (IC 50 = 3.68 ± 0.13 μM). Tyr-Asn-Leu-Lys-Glu-Arg-Tyr-Ala-Ala-Trp (IC 50 = 1.35 ± 0.23 μM) and Tyr-Asn-Arg-Leu-Pro-Glu-Leu (IC 50 = 3.42 ± 0.39 μM) exhibited the most potent ACE inhibitory activity. Ile-Ala-Asp-His-Phe-Leu showed the highest DPP-IV inhibitory activity with an IC 50 value of 610.1 ± 82.6 μM. All selected peptides were non-toxic, and most were non-allergenic according to in silico predictions. These results indicate the promising potential of bighead carp muscle hydrolysates as functional additives in foods and pharmaceuticals.

Scope The present study aims to assess the antidiabetic effect of Lactobacillus paracasei strain NL41 and its potential mechanisms in rats with type 2 diabetes mellitus (T2DM) induced by a high-fat diet and low-dose streptozotocin administration (HFD/STZ). Methods and results Eighteen Sprague-Dawley (SD) rats are randomly assigned to three groups: one control, one HFD/STZ model, and one HFD/STZ-Lactobacillus protection group with administration of strain NL41 for 12 weeks. Blood is collected for biochemical parameters analysis and tissue samples for histological analysis. Treatment with strain NL41 results in excellent blood glucose regulation and significantly decreases insulin resistance, and HbA1c, glucagon, and leptin levels, accompanied by remarkable improvement of dyslipidemia and oxidative stress status in the animals. Islets of Langerhans, liver, and kidney are significantly protected in the NL41-treated rats compared to the HFD/STZ-T2DM model rats. Histochemistry shows that strain NL41 inhibits beta-cell loss and alpha-cell expansion, indicating pancreatic islets as the targeted tissues for the primary ameliorative effect of the probiotic strain on HFD/STZ-T2DM rats. Crosstalk between the gut-liver and liver-pancreas endocrine axes is discussed. Conclusion Probiotic strain NL41 prevents HFD/STZ-T2DM by decreasing insulin resistance and oxidative stress status, and protecting beta-cell function.

Somatic mutation is associated with the occurrence, development and recurrence of oral cancer. Oral squamous cell carcinoma cell lines have been widely used in oral cancer research. So it's important to understand their genetic background, especially the mutation profile. Using the data of The Catalogue Of Somatic Mutations In Cancer (COSMIC), we analyzed the mutation characteristics of six tongue squamous cell carcinoma cell lines, identified the most common mutation cancer genes in oral squamous cell carcinoma, and compared these genes with those in colon cancer and lung cancer. Six tongue cancer cells have similar mutation spectrum in the coding region, of which C > T transition is the most common mutation, mainly located in the context of CpG dinucleotides. Profiles of mutation genes vary in different tongue cancer cell lines and TP53 is the most common mutation gene, followed by CDKN2A, KMT2D, and LRP1B. The 20 most common mutation genes in oral cancer are different from those in colon cancer and lung cancer, although TP53 is the most common mutation gene in these three cancers. The mutation characteristics of tongue cancer cell lines and mutation gene profiles of oral cancer provide valuable information for the study of oral cancers.

// Fenfang Wu 1, * , Di Wu 1, * , Yong Ren 1 , Chongyang Duan 2 , Shangwu Chen 1 , Anlong Xu 1, 3 1 Guangdong Province Key Laboratory for Pharmaceutical Functional Genes, College of Life Sciences, Sun Yat-Sen University, Guangzhou, Guangdong, 510006, People’s Republic of China 2 Department of Biostatistics, Southern Medical University, Guangzhou, Guangdong, 510515, People’s Republic of China 3 Beijing University of Chinese Medicine, Beijing, 100029, People’s Republic of China * These authors contributed equally to this work Correspondence to: Anlong Xu, email: lssxal@mail.sysu.edu.cn Keywords: acute promyelocytic leukaemia, Bayesian network meta-analysis, arsenic trioxide, all-trans retinoic acid, chemotherapy Received: March 07, 2016 Accepted: May 20, 2016 Published: June 17, 2016 ABSTRACT Acute promyelocytic leukemia (APL) is a curable subtype of acute myeloid leukemia. The optimum regimen for newly diagnosed APL remains inconclusive. In this Bayesian network meta-analysis, we compared the effectiveness of five regimens-arsenic trioxide (ATO) + all-trans retinoic acid (ATRA), realgar-indigo naturalis formula (RIF) which contains arsenic tetrasulfide + ATRA, ATRA + anthracycline-based chemotherapy (CT), ATO alone and ATRA alone, based on fourteen randomized controlled trials (RCTs), which included 1407 newly diagnosed APL patients. According to the results, the ranking efficacy of the treatment, including early death and complete remission in the induction stage, was the following: 1. ATO/RIF + ATRA; 2. ATRA + CT; 3. ATO, and 4. ATRA. For long-term benefit, ATO/RIF + ATRA significantly improved overall survival (OS) (hazard ratio = 0.35, 95%CI 0.15–0.82, p = 0.02) and event-free survival (EFS) (hazard ratio = 0.32, 95%CI 0.16–0.61, p = 0.001) over ATRA + CT regimen for the low-to-intermediate-risk patients. Thus, ATO + ATRA and RIF + ATRA might be considered the optimum treatments for the newly diagnosed APL and should be recommended as the standard care for frontline therapy.

Estrogen signaling influences the development and progression of ovarian tumors, but the underlying mechanisms are not well understood. In a previous study we demonstrated that impairment of estrogen receptor alpha (ERα)-mediated olfactomedin 4 (OLFM4) expression promotes the malignant progression of endometrioid adenocarcinoma, and we identified OLFM4 as a potential target of miR-486-5p. In this study we investigated the role of OLFM4 in ovarian serous adenocarcinoma. Ovarian serous adenocarcinoma tissues had reduced OLFM4 expression. Expression of OLFM4 was positively correlated with ERα expression, and estrogen (E2) treatment in ovarian cancer cells induced OLFM4 expression in an ERα-dependent manner. In contrast to ERα, miR-486-5p levels were inversely correlated with OLFM4 expression in ovarian serous adenocarcinoma. Ovarian cancer cells transfected with miR-486-5p mimics showed decreased OLFM4 mRNA expression, and ovarian cancer cells treated with E2 showed reduced cellular miR-486-5p levels. OLFM4 knockdown enhanced proliferation, migration, and invasion by ovarian cancer cells. Low expression of OLFM4 was also associated with high tumor FIGO stage and poor tumor differentiation. These results suggest OLFM4 is downregulated by miR-486-5p, which contributes to ovarian cancer tumorigenesis. Conversely, estrogen receptor signaling downregulates miR-486-5p and upregulates OLFM4 expression, slowing the development and progression of ovarian cancer.

The different potential for release of dipeptidyl peptidase IV (DPP-IV)-inhibitory peptides from bovine and caprine milk casein was assessed by in silico analysis and in vitro proteolytic assays. The former predicted a weakly higher potency for caprine casein than bovine as a precursor of DPP-IV-inhibitory peptides, although with markedly diverse potential for individual caseins. This was verified by protease hydrolysis; trypsin-treated casein hydrolysates (TCAHs) displayed the strongest bioactivity. Fractionation of caprine TCAHs revealed slightly, but significantly, higher inhibitory activity in peptides 5 kDa fraction, than their bovine counterparts. Through in silico trypsin hydrolysis and peptides synthesis, four novel caprine casein-derived DPP-IV-inhibitory peptides, including GPFPILV and HPINHR (half maximal inhibitory concentration = 163.7 ± 1.33 and 452.2 ± 7.15 μ m , respectively), were found. This study corroborates the weak superiority of caprine casein over bovine in release of DPP-IV-inhibitory peptides, and identifies several novel caprine casein-derived DPP-IV-inhibitory peptides.

Various nanomaterials have been extensively investigated for photothermal ablation of cancer cells because of their high photothermal conversion efficiency. However, the poor targeting specificity and low repeated heating efficiency of nanomaterials restrict their applications in the clinic. In this work, porous gelatin/iron oxide (Gel/Fe3O4) composite scaffolds were prepared by a facile ice particulate templating method for efficient and repeatable cancer cell ablation. Gel/Fe3O4 composite scaffolds showed controlled porous structure consisting of large pores and interconnecting small pores. The strong absorption in the near-infrared (NIR) region enabled the Gel/Fe3O4 composite scaffolds to elevate local temperature quickly under NIR laser irradiation. The composite scaffolds allowed cell adhesion and proliferation showing good biocompatibility. Cancer cells entrapped in the scaffolds could be efficiently killed during laser irradiation. Moreover, the therapeutic efficacy of Gel/Fe3O4 composite scaffolds could be enhanced by repeated laser irradiation treatment, which is important for clinical application because of the resistant and recurrent nature of cancer. The results indicated that the porous Gel/Fe3O4 composite scaffolds had good biocompatibility and excellent cancer cell ablation efficacy, which may provide an attractive way to use porous scaffolds for cancer therapy application.

Lactobacillus strains were screened for inhibition of dipeptidyl peptidase IV (DPP-IV) and α-glucosidase. Cell-free excretory supernatants (CFS) and cell-free extracts (CFE) of 21 strains were prepared, and most of them showed DPP-IV inhibition in a concentration-dependent manner, which started at the exponential phase and peaked at the stationary phase of the Lactobacillus . The CFS bioactivity was heat-resistant, stable to acid–alkali and glucosidase treatment. Trypsin treatment specifically elevated CFS's DPP-IV inhibition. Lactobacillus plantarum strains CFS displayed different protein patterns and trypsin sensitive bands on SDS-PAGE, demonstrating specific peptide-cleavage contributes to higher inhibition. Seven strains with highest DPP-IV inhibition also displayed α-glucosidase inhibitory activity, of which, three strains of L. plantarum and one of Lactobacillus brevis showed tolerance to simulated gastrointestinal tract conditions and high HT-29 cell adhesion. This initial report of lactobacilli secreting strong DPP-IV and α-glucosidase inhibition components will be beneficial for selection and application of anti-diabetic probiotics.

Probiotics are live microorganisms that when administered in adequate amounts, confer health benefits on the host. In this study, 13 strains of Bifidobacterium were isolated from three samples of breast-fed infant feces. The isolates were identified based on conservative gene sequencing and phylogenetic analysis. In vitro tests included survival under simulated gastrointestinal tract conditions, aggregation, hydrophobicity, intestinal epithelial cell adhesion, antimicrobial activity, and antibiotic resistance according to international guidelines for probiotics. The results suggest that B. bifidum, B. adolescentis, and B. breve had high adhesive ability compared with B. longum and B. catenulatum/B. pseudocatenulatum group strains. In particular, B. bifidum IF3-211 has a highest adhesion index (8273 ± 247 and 18,009 ± 1476 adhering bacteria per 100 HT-29 and Caco-2 cells, respectively), far higher than the two reference strains, B. lactis Bb12 and B. longum BBMN68. B. adolescentis IF1-11 showed highest autoaggregation (82.52 ± 0.24 %) and coaggregation (45.59 ± 4.16 %) with L. monocytogenes among isolates. In conclusion, B. bifidum IF3-211 and B. adolescentis IF1-11 showed promising characteristics as probiotic candidates that have good potential for application in food industry.

New dipeptidyl peptidase IV (DPP-IV)-inhibitory peptides from trypsin/chymotrypsin-treated goat milk casein hydrolysates were isolated and identified by two-dimensional silica thin-layer chromatography (2D-TLC) combined to nano LC-MS/MS. 2D-TLC with chloroform/methanol/25% ammonia (2:2:1) and n-butanol/acetic acid/water (4:1:1) as the first- and second-dimension eluents, respectively, in analytical and semipreparative scales, was set up and verified by reversed-phase high-performance liquid chromatography (RP-HPLC) to be feasible and efficient to separate the hydrolysates. Five new DPP-IV-inhibitory peptides, four relatively large oligopeptides (MHQPPQPL, SPTVMFPPQSVL, VMFPPQSVL, and INNQFLPYPY), and AWPQYL were identified, and INNQFLPYPY showed a notable IC50 value of 40.08 μM as an uncompetitive inhibitor. Interactive effects on DPP-IV inhibition were also observed among separated fractions and pure synthetic peptide mixtures with concentration-dependent activity. The study gives new insights into goat casein hydrolysates with identified DPP-IV-inhibitory peptides efficiently isolated by 2D-TLC, which provides a simple and cost-efficient separation process and is compatible with liquid chromatography-tandem mass spectrometry (LC-MS/MS) identification.

Engineering cartilage tissue by culturing chondrocytes in porous scaffolds is one promising method to repair or restore the functions of diseased cartilage. Hyaluronic aid (HA) was used in porous scaffolds or hydrogels to promote the proliferation of chondrocytes and synthesis of cartilage extracellular matrix (ECM). However, whether HA in scaffolds has a beneficial effect on chondrocytes remains uncertain, possibly due to the uncontrolled pore structure and inhomogeneous HA in scaffolds. In this study, homogeneous collagen/HA scaffolds with well-controlled and interconnected pore structure were prepared by suppression of polyion complex formation between collagen and HA and using ice particulates as porogen. The pore structure and mechanical property of collagen/HA scaffolds and collagen scaffolds could be well controlled. High molecular weight HA in collagen/HA scaffolds inhibited the cellular proliferation, synthesis of sulfated glycosaminoglycan and cartilage ECM, compared with the results of collagen scaffolds. This study should provide additional information on the effects of HA in porous scaffolds on the chondrocyte behaviour in 3D culture.

Highlights • Chloramphenicol treatment induced immunosuppression and severe tissue damage in amphioxus. • KEGG clustering showed that chloramphenicol and ampicillin treatment resulted in immunostimulation. • Chloramphenicol treatment induced a ∼3-fold decrease of eicosanoid levels. • Chloramphenicol and ampicillin treatment resulted in a 1.7-fold increase of eicosanoid levels. • Eicosanoids derived from arachidonic acid provide insights into the effect of chloramphenicol treatment., Chloramphenicol (Chl) is an effective antimicrobial agent widely used in veterinary medicine and commonly used in fish. Its use is restricted in the clinic because of adverse effects on the immune system and oxidative stress in mammals. However, the effects of Chl treatment on invertebrates remain unclear. Amphioxus, a basal chordate, is an ideal model to study the origin and evolution of the vertebrate immune system as it has a primary vertebrate-like arachidonic acid (AA) metabolic system. Here, we combined transcriptomic and lipidomic approaches to investigate the immune system and observe the oxygenated metabolites of AA to address the antibiotic effects on amphioxus. Tissue necrosis of the gill slits occurred in the Chl-treated amphioxus, but fewer epithelial cells were lost when treated with both Chl and ampicillin (Amp). The immune related pathways were dysregulated in both of the antibiotic treatment groups. The Chl alone treatment resulted in immunosuppression with down-regulation of the innate immune genes. In contrast, the Chl + Amp treatment resulted in immunostimulation to some extent, as shown by KEGG clustering. Furthermore, Chl induced a 3-fold reduction in the level of the eicosanoids, while the Chl + Amp treatment resulted in 1.7-fold increase of eicosanoid level. Thus in amphioxus, Amp might relieve the effects of the Chl-induced immune suppression and increase the level of eicosanoids from AA. Finally, the oxygenated metabolites from AA might be crucial to evaluate the effects of Chl treatment in animals.

In recent years, therapeutic peptides have garnered great interest in the pharmaceutical industry for the treatment of diabetes. Lactic acid bacteria (LAB) are an appealing vehicle for safe and convenient oral delivery of bioactive peptide and protein drugs. Exendin-4 (Exd4) is a glucagon-like protein-1 (GLP-1) receptor agonist that is considered an excellent therapeutic peptide drug for type 2 diabetes due to its longer-lasting bioactivity, resulting from resistance to dipeptidyl peptidase 4. We explored Lactococcus lactis with the nisin-controlled gene expression (NICE) system as an oral delivery system for recombinant (r) Exd4 peptide in situ. Heterologous expression and secretion of rExd4 by L. lactis NZ9000/pNZ8048-rExd4 were successful and efficient under the NICE system. In vitro treatment with rExd4 significantly enhanced insulin secretion of INS-1 cells and activated the PI3-K/AKT signal pathway with protein levels of AKT and p-AKT increasing 1.6- to 1.8-fold compared to negative controls, similar to the positive GLP-1 controls. INS-1 cells treated with rExd4 also showed enhanced proliferation and inhibited apoptosis, corresponding with the effects of the standard Exd4 and GLP-1 treatments. Our data suggest that the rExd4 secreted by L. lactis is a bioactive insulinotropic peptide and functional GLP-1 receptor agonist that enhances glucose-dependent insulin secretion and activates the PI3-K/AKT signal pathway; furthermore, it may be involved in improving proliferation and inhibiting apoptosis of INS-1 cells in in vitro treatments. Therefore, L. lactis producing rExd4 may potentially serve as a novel strategy for oral treatment of diabetes.

Porous scaffolds can be used to engineer three-dimensional (3D) tissues for tissue repair. Coculture of vascular endothelial cells and the cells of target tissue in porous scaffolds is promising to engineer vascularized tissue. However, it is difficult to induce regeneration of anisotropic tissues such as muscle and nerve that have well-aligned cells, blood vessels, and extracellular matrix (ECM) by using conventional porous scaffolds with homogeneous pore structures. In this study, we developed collagen porous scaffolds with parallel and concave microgrooves by using micropatterned ice template and freeze drying. Vascular endothelial cells and skeletal muscle myoblasts were cocultured on the microgrooved collagen scaffolds and control collagen scaffolds without micropattern structure. When the two types of cells were seeded at a proper ratio and concentration, the 3D microgrooved collagen scaffolds triggered spontaneous cell assembly into anisotropic muscle bundles with well-aligned tubule-like structure. Muscle cells were highly aligned in the tissue bundles, showing high expression of myosin heavy chain and incorporating the aligned tubule structure of vascular endothelial cells. Micropatterned porous scaffolds with microgrooves enabled engineering of anisotropic tissue with well-ordered tubules and could provide a platform for the study of cell assembly in micropatterned material environment.

Bifidobacteria are generally sensitive to oxidative stress caused by reactive oxygen species (ROS). To improve oxidative-stress tolerance, the superoxide dismutase (SOD) gene from Streptococcus thermophilus (StSodA) and the heme-dependent catalase (KAT) gene from Lactobacillus plantarum (LpKatL) were heterologously expressed in Bifidobacterium longum strain NCC2705. Three types of strain NCC2705 transformants were obtained: with transgenic SOD expression, with transgenic KAT expression, and with coexpression of the two genes. Intracellular expression of the genes and their functional role in oxidative-stress resistance were evaluated. In response to oxidative stress, B. longum NCC2705/pDP401-LpKatL (expressing LpKatL) and NCC2705/pDP-Kat-Sod (coexpressing LpKatL and StSodA) rapidly degraded exogenous H2O2 and the peroxides generated as a byproduct of aerobic cultivation, preventing oxidative damage to DNA and RNA. Individual expression of StSodA or LpKatL both improved B. longum NCC2705 cell viability. Survival rate of strain NCC2705 was further improved by combining SOD and KAT expression. The two enzymes played complementary roles in ROS-scavenging pathways, and coexpression led to a synergistic beneficial effect under conditions of intensified oxidative stress. Our results illustrate that heterogeneous expression of heme-dependent KAT and Mn(2+)-dependent SOD is functional in the B. longum oxidative-stress response, and synergistic protection is achieved when their expressions are combined.

Intestinal bifidobacteria benefit human health by promoting and modulating the gut flora, and boosting therapeutic efficiency for chronic metabolic diseases and cancer. Recently, Bifidobacterium adolescentis strains with high adhesion to intestinal epithelial cells were associated with induction of T-helper 17 (Th17) cells in humans and rodents. Here, two B. adolescentis strains with similar adhesive ability but different aggregation properties were investigated for specific immunoregulatory effects, including the underlying cellular pathway, on macrophage and T-regulatory (Treg)/Th17 axis activation in vitro and in the colon of dextran sodium sulfate (DSS)-colitis mice in vivo. In-vitro, the auto-aggregative B. adolescentis strain IF1-11 induced significantly higher IL-6 and lower IL-10 secretion from immune cells, and it induced abundant Th17 cells. The non-aggregating strain IF1-03 induced significantly higher IL-10, less IL-6 and a high proportion of Treg/Th17 cells compared to total T cells. In vivo, orally administered IF1-03 protected DSS-colitis mice via activation of dendritic cells or macrophages and skewing of Treg/Th17 cells, consistent with Treg cell induction in vitro. IF1-03 exopolysaccharides showed a functional recognition pattern similar to IF1-03 for IL-10 cytokine secretion and Treg cell-differentiation induction, both dependent on the toll-like receptor 2&ndash, ERK/p38 MAPK-signaling cascade for macrophage activation. We suggest that B. adolescentis exopolysaccharide-associated enterocyte adhesion/aggregation phenotypes determine strain-specific adaptive immune responses in the gut via the macrophage-regulated Treg/Th17 axis.

Controlled and local release of bioactive growth factors from porous scaffolds can provide an efficient strategy to control the complex and dynamic regulation of cellular processes in a three-dimensional microenvironment. In this study, hybrid scaffolds of collagen and poly(lactic- co-glycolic acid) microbeads were prepared by introducing insulin-releasing poly(lactic- co-glycolic acid) microbeads into collagen porous scaffolds. Insulin-incorporated poly(lactic- co-glycolic acid) microbeads of two distinct sizes, 19.4 ± 1.6 and 4.4 ± 0.9 µm, were used to prepare the hybrid scaffolds. The scaffolds had controlled pore structure, and the poly(lactic- co-glycolic acid) microbeads were well distributed on the pore walls of the scaffolds. The scaffolds had a lower initial burst and a more stable insulin release than did the free microbeads. Culture of human dermal fibroblasts in the hybrid scaffolds were affected the bioactivity of released insulin. The hybrid scaffold prepared with 19.4 ± 1.6 µm microbeads had a more linear release of insulin and a higher promotion effect on cell proliferation than did the other hybrid group and control scaffolds. The hybrid scaffold should be useful for skin tissue engineering.

Aberrant expression of high mobility group box 1 (HMGB1) is associated with tumor development and progression. The current study was conducted to evaluate the significance of HMGB1 immunostaining on cell block (CB) preparations in the diagnosis of neoplastic and preneoplastic lesions of the cervix. The CBs were prepared from 157 residual liquid-based gynecologic cytology specimens which were collected from women whose cervical lesions had been confirmed by histopathology. The expression of HMGB1 and p16INK4A (p16) was visualized by immunocytochemistry on the CB preparations, and the association of their expression patterns was correlated with the severity of cervical lesions. HeLa cells were used as positive control. HMGB1 expression was observed in dysplastic and neoplastic cells and increased along with the progression of cervical neoplasia. The rates of positive staining for HMGB1 in cervical intraepithelial neoplasia 1 (CIN-1), CIN-2, CIN-3, and invasive squamous cell carcinomas (ISCCs) were 69.4, 96.9, 100.0, and 100.0%, respectively. The differences between positive rates of patients with chronic cervicitis and various CINs as well as ISCCs were significant (P

As a major class of pattern-recognition receptors, Toll-like receptors (TLRs) play a critical role in defense against invading pathogens. Increasing evidence demonstrates that, in addition to infection, TLRs are involved in other important pathological processes, such as tumorigenesis. Activation of TLRs results in opposing outcomes, pro-tumorigenic effects and anti-tumor functions. TLR signaling can inhibit apoptosis and promote chronic inflammation-induced tumorigenesis. TLR activation in tumor cells and immune cells can induce production of cytokines, increase tumor cell proliferation and apoptosis resistance, promote invasion and metastasis, and inhibit immune cell activity resulting in tumor immune escape. In contrast, the engagement of other TLRs directly induces growth inhibition and apoptosis of tumor cells and triggers activation of immune cells enhancing anti-tumor immune responses. Thus, the interpretation of the precise function of each TLR in tumors is very important for targeting TLRs and using TLR agonists in tumor therapy. We review the role of TLR signaling in tumors and discuss the factors that affect outcomes of TLR activation.

Background Proteinaceous bioactive substances and pharmaceuticals are most conveniently administered orally. However, the facing problems are the side effects of proteolytic degradation and denaturation in the gastrointestinal tract. In recent years, lactic acid bacteria (LAB) have been verified to be a promising delivery vector for susceptible functional proteins and drugs. KiSS-1 peptide, a cancer suppressor, plays a critical role in inhibiting cancer metastasis and its activity has been confirmed by direct administration. However, whether this peptide can be functionally expressed in LAB and exert activity on cancer cells, thus providing a potential alternative administration manner in the future, has not been demonstrated. Results A recombinant Lactococcus lactis strain NZ9000-401-kiss1 harboring a plasmid containing the gene of the tumor metastasis-inhibiting peptide KiSS1 was constructed. After optimization of the nisin induction conditions, the recombinant strain efficiently secreted KiSS1 with a maximum detectable amount of 27.9 μg/ml in Dulbecco’s Modified Eagle medium. The 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide and would healing assays, respectively, indicated that the secreted KiSS1 peptide remarkably inhibited HT-29 cell proliferation and migration. Furthermore, the expressed KiSS1 was shown to induce HT-29 cell morphological changes, apoptosis and reduce the expression of matrix metalloproteinase 9 (MMP-9) at both mRNA and protein levels. Conclusions A recombinant L. lactis NZ9000-401-kiss1 successfully expressing the human kiss1 was constructed. The secreted KiSS1 peptide inhibited human HT-29 cells’ proliferation and migration probably by invoking, or mediating the cell-apoptosis pathway and by down regulating MMP-9 expression, respectively. Our results suggest that L. lactis is an ideal cell factory for secretory expression of tumor metastasis-inhibiting peptide KiSS1, and the KiSS1-producing L. lactis strain may serve as a new tool for cancer therapy in the future.

Bifidobacterium longumsubsp.longumBBMN68, an anaerobic probiotic isolated from healthy centenarian faeces, shows low oxygen (3 %, v/v) tolerance. To understand the effects of oxidative stress and the mechanisms protecting against it in this strain, a proteomic approach was taken to analyse changes in the cellular protein profiles of BBMN68 under the following oxygen-stress conditions. Mid-exponential phase BBMN68 cells grown in MRS broth at 37 °C were exposed to 3 % O2for 1 h (I) or 9 h (II), and stationary phase cells were subjected to 3 % O2for 1 h (III). Respective controls were grown under identical conditions but were not exposed to O2. A total of 51 spots with significant changes after exposure to oxygen were identified, including the oxidative stress-protective proteins alkyl hydroperoxide reductase C22 (AhpC) and pyridine nucleotide-disulfide reductase (PNDR), and the DNA oxidative damage-protective proteins DNA-binding ferritin-like protein (Dps), ribonucleotide reductase (NrdA) and nucleotide triphosphate (NTP) pyrophosphohydrolases (MutT1). Changes in polynucleotide phosphorylase (PNPase) plus enolase, which may play important roles in scavenging oxidatively damaged RNA, were also found. Following validation at the transcriptional level of differentially expressed proteins, the physiological and biochemical functions of BBMN68 Dps were further proven byin vitroandin vivotests under oxidative stress. Our results reveal the key oxidative stress-protective proteins and DNA oxidative damage-protective proteins involved in the defence strategy of BBMN68 against oxygen, and provide the first proteomic information toward understanding the responses ofBifidobacteriumand other anaerobes to oxygen stress.

Toll-like receptors are an important family of pattern recognition receptors. They recognize microbial conserved components and trigger protective responses to the invading pathogens, which constitute a major part of the innate immune system. Toll-like receptors are mainly expressed in immune cells. The current evidences demonstrate that Toll-like receptors are present in some epithelial cells and epithelium derived tumor cells. The expression of Toll-like receptors in these cells is related to infection and inflammation, and tumor progression as well. Genital mucosal epithelium is the first line in defense of microorganism invasion in the female reproductive tract. Toll-like receptors expressed in the genital tract have been implicated in many aspects of reproductive physiology and pathology in the female. In the current review, we will focus on the expression of Toll-like receptors in the female genital mucosa and its association with anti-infection immunity and tumorigenesis.

HKABF, Hippocampus kuda antibacterial factor, is a putative antimicrobial peptide identified from the cDNA library of the brooding pouch of H. kuda Bleeker. The putative mature peptide, fused with a 6His-tag, an anionic peptide (SAPLNPDNID) and to an enterokinase cleavage site was expressed in Pichia pastoris. The mature HKABF peptide, released by enterokinase cleavage, showed antimicrobial activity against Staphylococcus aureus and Staphylococcus saprophyticus at low concentrations (0.5–2.5 μg/ml). The recombinant mature peptide was heat-stable and had low hemolytic activity. Circular dichroism (CD) spectroscopy data indicates its classification as a CSαβ-type antimicrobial peptide. This study serves as an impetus for the production and further research of this antimicrobial peptide.

A T-1-conotoxin, lt5d, was purified and characterized from the venom of vermivorous hunting cone snails Conus litteratus . The complete amino acid sequence of lt5d (DCCPAKLLCCNP) has been determined by Edman degradation. With two disulfide bonds, the calculated average mass is 1274.57 Da, which is confirmed by MALDI-TOF mass spectrometry (average mass 1274.8778). Under whole cell patch-clamp mode, lt5d inhibits tetrodotoxin-sensitive sodium currents on adult rat dorsal root ganglion neurons, but has no effects on tetrodotoxin-resistant sodium currents. The inhibition of TTX-sensitive sodium currents by lt5d was found to be concentration-dependent with the IC 50 value of 156.16 nM. Thus, this is the first T-superfamily conotoxin identified to block TTX-sensitive sodium channels.

Human leukocyte antigen (HLA) class II alleles have been associated with an increased or decreased risk of developing cervical cancer through infection with oncogenic forms of human papillomavirus (HPV). To verify whether HLA class II allelic polymorphism is related to cervical cancer in China, 133 cervical cancers and 98 healthy controls were analyzed for HLA typing. Our results showed that DPB1*1301 allele frequency was significantly higher in the cervical cancers compared with healthy controls (OR, 3.793; p = 0.002; Pc = 0.04). A significant relationship was found between DRB1*150101-DQB1*0602 haplotype (OR, 0.180; p < 0.0001; Pc < 0.003), DRB1*070101-DQB1*0201 haplotype (OR, 0.110; p = 0.001; Pc = 0.03), and decreased risk for cervical cancer. Similar tendencies were observed for DRB1*150101-DQB1*0602 haplotype with HPV16 positive cervical cancers (OR, 0.182; p = 0.001; Pc = 0.021), and for DRB1*070101-DQB1*0201 haplotype (OR, 0.144; p =0.003; Pc = 0.063). These results indicate that HLA-DPB1*1301 may confer susceptibility to cervical cancer, and the haplotypes DRB1*150101-DQB1*0602 and DRB1*070101-DQB1*0201 may contribute to the resistance to the development of cervical cancer among Chinese women. The study suggests that specific HLA class II alleles and haplotypes may influence the immune response to specific HPV-encoded epitopes and affect the risk of cervical cancer in a Chinese population from an area with a high incidence of this neoplasia.

Engineering of cartilage tissue in vitro using porous scaffolds and chondrocytes provides a promising approach for cartilage repair. However, nonuniform cell distribution and heterogeneous tissue formation together with weak mechanical property of in vitro engineered cartilage limit their clinical application. In this study, gelatin porous scaffolds with homogeneous and open pores were prepared using ice particulates and freeze-drying. The scaffolds were used to culture bovine articular chondrocytes to engineer cartilage tissue in vitro. The pore structure and mechanical property of gelatin scaffolds could be well controlled by using different ratios of ice particulates to gelatin solution and different concentrations of gelatin. Gelatin scaffolds prepared from ≥70% ice particulates enabled homogeneous seeding of bovine articular chondrocytes throughout the scaffolds and formation of homogeneous cartilage extracellular matrix. While soft scaffolds underwent cellular contraction, stiff scaffolds resisted cellular contraction and had significantly higher cell proliferation and synthesis of sulfated glycosaminoglycan. Compared with the gelatin scaffolds prepared without ice particulates, the gelatin scaffolds prepared with ice particulates facilitated formation of homogeneous cartilage tissue with significantly higher compressive modulus. The gelatin scaffolds with highly open pore structure and good mechanical property can be used to improve in vitro tissue-engineered cartilage.

Adhesion to host cells is considered important for Lactobacillus plantarum as well as other lactic acid bacteria (LAB) to persist in human gut and thus exert probiotic effects. Here, we sequenced the genome of Lt. plantarum strain NL42 originating from a traditional Chinese dairy product, performed comparative genomic analysis and characterized a novel adhesion factor. The genome of NL42 was highly divergent from its closest neighbors, especially in six large genomic regions. NL42 harbors a total of 42 genes encoding adhesion-associated proteins; among them, cwaA encodes a protein containing multiple domains, including five cell wall surface anchor repeat domains and an LPxTG-like cell wall anchor motif. Expression of cwaA in Lactococcus lactis significantly increased its autoaggregation and hydrophobicity and conferred the new ability to adhere to human colonic epithelial HT-29 cells by targeting cellular surface proteins and not carbohydrate moieties, for CwaA adhesion. In addition, the recombinant Lc. lactis inhibited adhesion of Staphylococcus aureus and Escherichia coli to HT-29 cells, mainly by exclusion. We conclude that CwaA is a novel adhesion factor in Lt. plantarum and a potential candidate for improving the adhesion ability of probiotics or other bacteria of interest.

Risk factors of the development of distant metastasis in primary node-negative breast cancer patients are heterogeneous. Identification of patients at high risk of early distant metastasis is of important clinical significance. In the current study, using the already published datasets, we develop a gene signature that can robustly predict early distant metastasis for patients with primary node-negative breast cancer. We identified a 39-gene signature, which were associated with distant metastasis and shorter distant metastasis free survival (DMFS) in node-negative breast cancers. Using the survival prediction analysis method in BRB-Array tools, this signature can stratify patients into early- and late- distant metastasis subgroups with different DMFS in VDX training dataset (AUC=0.734, P < 0.01). And we further validated the reliability of the prognostic value of this 39-gene signature in another two independent breast cancer cohorts (NKI dataset, AUC=0.642, P

Preparation of three-dimensional (3D) micropatterned porous scaffolds remains a great challenge for engineering of highly organized tissues such as skeletal muscle tissue and cardiac tissue. Two-dimensional (2D) micropatterned surfaces with periodic features (several nanometers to less than 100 μm) are commonly used to guide the alignment of muscle myoblasts and myotubes and lead to formation of pre-patterned cell sheets. However, cell sheets from 2D patterned surfaces have limited thickness, and harvesting the cell sheets for implantation is inconvenient and can lead to less alignment of myotubes. 3D micropatterned scaffolds can promote cell alignment and muscle tissue formation. In this study, we developed a novel type of 3D porous collagen scaffolds with concave microgrooves that mimic muscle basement membrane to engineer skeletal muscle tissue. Highly aligned and multi-layered muscle bundle tissues were engineered by controlling the size of microgrooves and cell seeding concentration. Myoblasts in the engineered muscle tissue were well-aligned and had high expression of myosin heavy chain and synthesis of muscle extracellular matrix. The microgrooved collagen scaffolds could be used to engineer organized multi-layered muscle tissue for implantation to repair/restore the function of diseased tissues or be used to investigate the cell-cell interaction in 3D microscale topography.

Yak milk casein derived from Qula, a traditional Tibetan acid curd cheese, was hydrolyzed by six commercially available proteases (Trypsin, Pepsin, Alcalase, Flavourzyme, Papain and Neutrase). These hydrolysates were assayed for their inhibitory activity of Angiotensin-I-converting enzyme (ACE). The hydrolysates obtained by Neutrase from Bacillus amyloliquefaciens showed the highest ACE inhibitory activity. The IC50 value of Neutrase-hydrolysate was 0·38 mg/ml. The hydrolysate obtained by Neutrase was further separated by consecutive ultra-filtration with 10 kDa and then with 6 kDa molecular weight cut-offs into different permeated parts and fractionated by gel filtration chromatography with a Sephadex G-25 column. The active fraction was subjected to RP-HPLC, in which five peaks were purified and identified. Amino acid sequence analysis confirmed that the peptides and origins were as follows: YQKFPQY (αs2-CN; f89–95), LPQNIPPL (β-CN; f70–77), SKVLPVPQK (β-CN; f168–176), LPYPYY (κ-CN; f56–61) and FLPYPYY (κ-CN; f55–61). Their amino acid sequences matched well with those of known bioactive peptides from bovine casein. The results indicated that yak milk casein could be a resource to generate antihypertensive peptides and be used as multifunctional active ingredients for many value-added functional foods as well as a traditional food protein.