ENPP2 inhibitor improves proliferation in AOM/DSS-induced colorectal cancer mice via remodeling the gut barrier function and gut microbiota composition.

In our previous multicenter study, we delineated the inherent metabolic features of colorectal cancer (CRC). Therein, we identified a member of the ectonucleotide pyrophosphatase/ phosphodiesterase family (ENPP2) as a significant differential metabolite of CRC. In this study, the role of ENPP2 in CRC has been demonstrated using established in vitro and in vivo models including ENPP2 gene knockdown, and use of the ENPP2 inhibitor, GLPG1690. We found that CRC proliferation was decreased after either ENPP2 gene knockdown or use of ENPP2 inhibitors. We further evaluated the role of GLPG1690 in AOM/DSS-induced CRC mice via intestinal barrier function, macrophage polarization, inflammatory response and microbial homeostasis. Results of immunofluorescence staining and Western blotting showed that GLPG1690 can restore gut-barrier function by increasing the expression of tight junction proteins, claudin-1, occludin and ZO-1. M2 tumor-associated macrophage polarization and colonic inflammation were attenuated after treatment with GLPG1690 using the Azoxymethane/Dextran Sodium Sulfate (AOM/DSS) model. Moreover, 16 S rDNA pyrosequencing and metagenomic analysis showed that GLPG1690 could alleviate gut dysbiosis in mice. Furthermore, administration of GLPG1690 with antibiotics as well as fecal microbiota transplantation assays demonstrated a close link between the efficacy of GLPG1690 and the gut microbiota composition. Finally, results of metabolomic analysis implicated mainly the gut microbiota-derived metabolites of aromatic amino acids in CRC progression. These findings may provide novel insights into the development of small-molecule ENPP2 inhibitors for the treatment of CRC.

Factors Associated With Neurosyphilis in Patients With Syphilis Treatment Failure: A Retrospective Study of 165 HIV-Negative Patients.

Background: In recent years, the incidence of syphilis has increased year by year. Our study is to explore the risk factors for the development of neurosyphilis in patients who failed syphilis treatment. Methods: A total number of 165 patients with complete medical records and who agreed to undergo lumbar puncture were divided into 47 neurosyphilis cases and 118 non-neurosyphilis cases according to the diagnostic criteria of neurosyphilis, and the differences in clinical characteristics and laboratory features between the two groups were analyzed. Significant variables were entered into multivariable logistic regression models. Results: (1) There were statistical differences (p < 0.05) between the neurosyphilis (NS) group and the non-neurosyphilis (NNS) group in terms of the higher proportion of male and serum rapid plasma reagin (RPR) > 1:32 and the elevated cerebrospinal fluid white blood cell (CSF WBC) and CSF protein in the neurosyphilis group compared with the non-neurosyphilis group. (2) Male gender, serum RPR titers >1:32 at lumbar puncture, CSF WBC >8 × 106/L were significantly associated with neurosyphilis. Conclusion: For patients who have failed syphilis treatment, lumbar puncture should be performed to exclude neurosyphilis, to enable early diagnosis and treatment, and to prevent irreversible damage of neurosyphilis, especially if the patient is male and has a serum RPR>1:32 and elevated CSF WBC at lumbar puncture, which are risk factors for neurosyphilis.

Extraction, isolation and identification of four phenolic compounds from Pleioblastus amarus shoots and their antioxidant and anti-inflammatory properties in vitro.

Pleioblastus amarus (P. amarus) shoots, belong to the grass family Gramineae, a traditional green vegetable in China, are rich in nutritional properties, and can provide various health benefits. This study isolated four compounds, namely (1-4), 3-O-coumaroylquinic acid (1), 3-O-feruloylquinic acid (2), 4-O-feruloylquinic acid (3), and 5-O-feruloylquinic acid (4) from Pleioblastus amarus shoots for the first time. The structures of the extracted compounds were determined using detailed spectroscopic (1D/2D NMR), high resolution electrospray ionization mass spectrometry (HR-ESI-MS), and infrared (IR) spectroscopy. The antioxidant capacity of 3-O-feruloylquinic acid (2) was stronger than that of the other compounds, while it also exhibited anti-inflammatory activity, significantly restricting the release of nitric oxide (NO) by lipopolysaccharide (LPS)-induced RAW 264.7 cells, displaying an inhibitory rate of 60.92 percent at a concentration of 400 µg/mL. Furthermore, 3-O-feruloylquinic acid (2) inhibited interleukin-1ß (IL-1ß), interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and nuclear factor-κB (NF-κB) expression and may be useful for developing novel antioxidant and anti-inflammatory substances.

Comparative transcriptomic analysis of Lactiplantibacillus plantarum RS66CD biofilm in high-salt conditions and planktonic cells.

BACKGROUND: Lactiplantibacillus plantarum (L. plantarum), a dominant strain in traditional fermented foods, is widely used in fermentation industry because of its fast acid production. However, L. plantarum is easily inactivated due to acidity, high temperature and other factors. The formation of biofilm by bacteria can effectively increase environmental tolerance. Therefore, it is important to improve the environmental tolerance of L. plantarum by studying its biofilm formation conditions and regulatory mechanisms. METHODS: After determining a suitable NaCl concentration for promoting biofilm formation, L. plantarum was grown with 48 g L-1 NaCl. Differential gene expressions in L. plantarum biofilm vs. planktonic cells were analyzed using RNA sequencing and validated using qPCR. RESULT: L. plantarum RS66CD biofilm formation formed highest amount of when grown at 48 g L-1 NaCl. Altogether 447 genes were up-regulated and 426 genes were down-regulated in the biofilm. KEGG pathway analysis showed that genes coding for D-Alanine metabolism, peptidoglycan biosynthesis, two-component system, carbon metabolism, bacterial secretion system, lysine biosynthesis and fatty acid metabolism were crucial for biofilm formation. In addition, eight other genes related to biofilm formation were differentially expressed. Our results provide insights into the differential gene expression involved in biofilm formation, which can help to reveal gene regulation during L. plantarum biofilm formation.

Isolation and identification of the spoilage microorganisms in Sichuan homemade Paocai and their impact on quality and safety.

The spoilage microbiology in Paocai (fermented vegetables) affects not only the quality of this popular traditional Chinese food but also its safety. This study aimed to isolate and identify the microorganisms commonly leading to spoilage in homemade Paocai from the Sichuan region and, further, to scientifically assess the impact of these microorganisms on product quality and safety. Seven putrid Paocai samples were collected from 7 families in different Sichuan cities. From these, 45 strains were isolated by means of a nutrient agar medium and rose bengal agar. All of the 22 strains (5 fungi and 17 bacteria) with different colonial morphologies and corruption phenomenon in Paocai were determined 16S rDNA/18S rDNA gene sequences. Bacteria were identified as Bacillus spp. (16 strains) and Staphylococcus saprophyticus (1 strain), while the 5 fungi were identified as Candida spp. (3 strains), Kodamaea ohmeri (1 strain), and Geotrichum candidum (1 strain). Based on the results of identification, 7 representative strains of different species were determined as the spoilage characteristics in paocai. All the representative strains metabolized to produce nitrite. Strain SPF21 (K. ohmeri) has a particularly serious impact on the crunchiness of Paocai; however, strains SPF19 (Bacillus subtilis) and SPF21 have the greatest influence on its color. All five of the fungi were seen to form a film on the surface of the Paocai, with SPF5 (G. candidum) exerting the most extreme influence. The growth characteristics in the broth showed that all the representative strains investigated metabolized most of the carbohydrates and were able to tolerate the salinity and acidity of the ordinary homemade Paocai. Moreover, these strains were found to have obvious impacts on the volatile components of Paocai, especially SPF2 and SPF8, with higher dimethyl disulfide and dimethyl trisulfide, which were found to have a pungent odor when highly concentrated.

[Variation of BDNF mRNA on focalcerebral ischemia reperfusion injury in rats with notogisenoside-Rg1].

OBJECTIVE: To study the effect of notoginsenoside-Rg, on expression of BDNF mRNA (brain-derived neurotrophic factor messenger ribonucleic acid) in cerebrum cortex after MCAO/R (middle cerebral artery occlusion reperfusion) injury in rat by real-time quantitative polymerase chain reaction. METHODS: 36 SD male rats were randomly divided into MCAO/R model group, notogisenoside-Rg1 therapy group (100 mg/kg) and the positive medicine control group (nimodipine 1 mg/kg). The MCAO/R model were made by thread-occluded method. The four rats were randomly taken from each groups and were killed to be breaken out brain tissues as specimens after which were treated with medicine in 1,3, 5 days. Total RNA was isolated from cerebrum cortex. Specific oligonucleotide primers of BDNF mRNA gene fragments were amplificated by RT-PCR to construct recombinant plasmid and sequence. To dilute recombinant plasmid didploidly and a quantitative standard curve was completed. Taqman fluorogenic quantitative RT-PCR (FQ-PCR) was set up to detect the BDNF mRNA variety of cerebrum cortex. RESULTS: Compared with the model group and the postive control group, notogisenoside-Rg1 treated groups could obviously improve some nervous deficit symptoms in the cerebral ischemia and increase BDNF mRNA amount that in the cerebrum cortex at different period after rat MCAO/R injury. CONCLUSION: Notoginsenoside-Rg1 can promote to generating internal BDNF protein in brain by up-regulation the expression of BDNF mRNA amount in the cerebrum cortex. BDNF bind in TrkB, which can give rise to protective effects for ischemic neurons by generating corresponding domino effect molecules. Accordingly it can be as a therapy method in cerebral ischemia injury.

Syphilis Infection Differentially Regulates the Phenotype and Function of γδ T Cells in HIV-1-Infected Patients Depends on the HIV-1 Disease Stage.

A rapidly escalating outbreak of syphilis infection has been affected men who have sex with men, particularly those with HIV-1 infection. γδ T cells are unconventional immune cells with two main subsets, Vδ1 T cells and Vδ2 T cells, which possess a combination of innate and adaptive immune features allowing them against HIV-1. However, whether syphilis infection affects the phenotype and function of γδ T cells in HIV-1-infected patients remains unclear, especially in acute HIV-1 infection (AHI). In this study, we enrolled 57 HIV-1-infected patients (24 with HIV-1 infection only and 33 coinfected with syphilis) from an acute HIV-1-infected cohort in Beijing (PRIMO). A comprehensive analysis of γδ T-cell phenotype and function was performed by flow cytometry. We found syphilis coinfection could reverse the imbalance of Vδ1/Vδ2 ratio in AHI. Syphilis infection results in decreased γδ T-cell activation in AHI, but increased γδ T-cell activation in chronic HIV-1 infection (CHI). Moreover, patients with CHI had larger numbers of IL-17-producing γδ T cells than those with AHI, regardless of syphilis status. Thus, syphilis affected the γδ T-cell immune response differently in patients depending on the stages of HIV-1 disease. In addition, the percentage of IL-17-producing γδ T cells was positively correlated with the percentage of neutrophils. These results suggest that the γδ T-cell/IL-17/neutrophil axis is involved in HIV-1 pathogenesis and disease progression. Taken together, our observations provide new insight into the roles of γδ T cells in immunopathogenesis of syphilis and HIV-1 coinfection, particularly during AHI, and our findings may be helpful for the prevention of syphilis and other sexually transmitted infections and highlight the great significance on the remedy of patients coinfected with HIV-1.

The expansion of autologous adipose-derived stem cells in vitro for the functional reconstruction of nasal mucosal tissue.

BACKGROUND: I t is established that adipose-derived stem cells (ADSCs) produce and secrete cytokines/growth factors that antagonize mucosal injury. However, the exact molecular basis underlying the treatment effects exerted by ADSCs is ill understood, and whether ADSCs cooperate with adipose tissue particles to improve mucosal function in patients with empty nose syndrome (ENS) has not been explored. We investigated the impact of ADSCs on nasal mucosa, the associated mechanisms, and their use in the treatment of patients with ENS. RESULTS: The nasal endoscope and mucociliary clearance assessments were significantly improved (P < 0.05) in patients with (n = 28) and without (n = 2) a rudimentary turbinate that received ADSCs combined with fat granules transplantation. Patients experienced a significant improvement in nasal obstruction and nasal mucociliary clearance after nasal turbinate angioplasty (P < 0.05). H&E staining, Masson's staining, and AB-PAS staining confirmed that inflammation was significantly reduced, collagenous fibers became aligned, fewer deposits were observed, and the mucosal proteins generated from caliciform cells increased following treatment. After a 14-day incubation period, ADSCs developed a polygonal cobblestone shape characteristic of human epithelial cells. Furthermore, immunohistochemical analysis revealed the presence of epithelial markers such as cytokeratin-7, and cytokeratin-19. Western blot analysis showed the presence of specific epithelial cell markers including cytokeratin-7, cytokeratin-14 and cytokeratin-19 in these epithelial like cells (ELC); these markers had low expression levels of ADSCs. CONCLUSIONS: The reconstruction of mucosal function by nasal turbinate angioplasty combined with ADSCs and autologous adipose tissue particle transplantation significantly improved the symptoms of patients with ENS. This is a new procedure that will improve mucosal restoration treatment options in patients with ENS. Furthermore, we undertook preliminary explorations of the underlying mechanisms involved, and found that transplantation of ADSCs could induce epithelial cells to improve mucosa function in patients with ENS in the micro-environment of injection areas.

Protective effect of crocetin on hemorrhagic shock-induced acute renal failure in rats.

Multiple organ failure is a common outcome of hemorrhagic shock followed by resuscitation, and the kidney is one of the prime target organs involved. The main objective of the study was to evaluate whether crocetin, a natural product from Gardenia jasminoides Ellis, has beneficial effects on renal dysfunction caused by hemorrhagic shock and resuscitation in rats. Anesthetized rats were bled to reduce mean arterial blood pressure to 35 (SD, 5) mmHg for 60 min and then were resuscitated with their withdrawn shed blood and normal saline. Crocetin was administered via the duodenum at a dose of 50 mg/kg 40 min after hemorrhage. The increase in creatinine and blood urea nitrogen was significantly reduced at 2 h after hemorrhage and resuscitation in crocetin-treated rats. The increases in renal nitric oxide, tumor necrosis factor α, and interleukin 6 were also attenuated by crocetin. Hemorrhagic shock resulted in a significant elevation in malondialdehyde production and was accompanied by a reduction in total superoxide dismutase activity, activation of nuclear factor κB, and overexpression of inducible nitric oxide synthase. These changes were significantly attenuated by crocetin at 2 h after resuscitation. These results suggested that crocetin blocks inflammatory cascades by inhibiting production of reactive oxygen species and restoring superoxide dismutase activity to ameliorate renal dysfunction caused by hemorrhage shock and resuscitation.

Effect of crocetin on blood pressure restoration and synthesis of inflammatory mediators in heart after hemorrhagic shock in anesthetized rats.

Crocetin, a constituent of saffron, has been shown not only to prevent reactive oxygen species-induced hepatotoxicity and genotoxicity but also to increase whole-body oxygen consumption and survival. The present study was to determine whether crocetin has beneficial effects on cardiac injury caused by hemorrhagic shock and resuscitation in rats. Anesthetized rats were bled to reduce mean arterial pressure (MAP) to 35 +/- 5 mmHg for 60 min and then resuscitated with their withdrawn shed blood and isotonic sodium chloride solution. Crocetin was administered via the duodenum at 50 mg/kg 40 min after bleeding. We investigated MAP, serum creatine kinase activity, the activity of nuclear factor-kappaB, iNOS, and total superoxide dismutase (T-SOD), as well as levels of NO, malondialdehyde, TNF-alpha, and IL-6 in the heart at 2 h postresuscitation. Compared with control group, crocetin significantly increased MAP from 10 min after administration to the end of the protocol except the period between 75 and 90 min after initial bleeding, whereas serum creatine kinase activity was dramatically decreased at 2 h postresuscitation. Myocardial nuclear factor-kappaB activity, iNOS activity, NO, malondialdehyde, TNF-alpha, and IL-6 were significantly elevated, whereas T-SOD activity was suppressed in the control group if compared with those of sham animals. These parameters tended to be normalized in rats administered crocetin. These results suggest that crocetin blocks inflammatory cascades by inhibiting reactive oxygen species production and preserving T-SOD activity to ameliorate the cardiac injury caused by hemorrhage/resuscitation.