Gastric bacteria as potential biomarkers for the diagnosis of atrophic gastritis.

BACKGROUND: Identification of the risk factors for atrophic gastritis (AG) and prevention of further deterioration of the gastritis are effective approaches to reduce the incidence of gastric cancer. Previous studies found that dysbiosis has been implicated in a wide range of diseases, while the role of gastric bacteria as a biomarker for AG has not been explored. METHODS AND RESULTS: Gastric juices from cases with non-atrophic gastritis (NAG) and AG were collected for investigation of bacterial composition and function. The ß-diversity of microbiota exhibited a significant reduction in AG samples compared with that in NAG samples. Differential abundance analysis revealed that a total of 23 predicted species changed their distributions; meanwhile, all obligate anaerobic bacteria with a relatively high abundance lowered their contents in AG samples. Additionally, the correlation analysis indicated a clear shift in bacterial correlation pattern between the two groups. Functional interrogation of the gastric microbiota showed that bacterial metabolisms associated with enzyme families, digestive system, and endocrine system were downregulated in AG samples. The compositional dissection of "core microbiota" exhibited that oral pathogens, including Porphyromonas gingivalis, Campylobacter gracilis, and Granulicatella elegans, were magnified in AG samples, suggesting that oral diseases may be a trigger factor for early exacerbation of gastritis. Then, the differentially expressed bacteria were used as diagnostic biomarkers for the random forest classifier model for group prediction. CONCLUSIONS: The results showed that bacterial biomarkers could distinguish AG patients from NAG cases with an accuracy of 90% at the genus level.

Habitat use of the white-headed langurs in limestone forest of Southwest Guangxi, China: Seasonality and group size effects.

Understanding how animals cope with habitat-specific environmental factors can assist in species conservation management. We studied the habitat use of four groups (two large and two small groups) of white-headed langurs (Trachypithecus leucocephalus) living in the forest of southwest Guangxi, China between September 2016 and February 2017 via instantaneous scan sampling. Our results showed that the langurs primarily used hillsides (55.91% ± 6.47%), followed by cliffs (29.70% ± 5.48%), hilltops (7.26% ± 3.55%), flat zones (6.99% ± 6.58%), and farmlands (0.14% ± 0.28%). The langurs moved most frequently on hillsides (49.35% ± 6.97%) and cliffs (35.60% ± 9.17%). The hillsides were more frequently used (66.94% ± 7.86%) during feeding, and the langurs increased the use of hilltops during the rainy season, and the use of cliffs in the dry season. The langurs frequently rested on hillsides (49.75% ± 8.16%) and cliffs (38.93% ± 8.02%). The larger langur group used cliffs more frequently when moving and resting, whereas the small langur group used hillsides more frequently while resting. Langurs in all groups avoided the flat zones for feeding. Their use of habitat reflected the balancing of foraging needs, thermoregulation, and predator avoidance. We conclude that the ecological factors are determinants of habitat use for white-headed langurs. Our findings suggest that conservation efforts should focus on protecting the vegetation on the hillsides and restoring the vegetation on the flat zones.

The toxicity of perfluorodecanoic acid is mainly manifested as a deflected immune function.

BACKGROUND: Perfluorodecanoic acid (PFDA) is a type of perfluoroalkyl acid (PFAA). PFDA has toxicity similar to dioxin; its effect on the body is not through a single target or a single pathway. However, the mechanism at the global level is still unclear. METHODS AND RESULTS: We treated mice with PFDA and characterized the global changes in gene expression in the liver using microarray analyses. The enriched KEGG pathways and GO analyses revealed that PFDA greatly affected the immune response, which was different from the response of gastric cells previously studied. As a proof of principle, the expressions of IL-1ß and IL-18 were both decreased after PFDA treatment, and qRT-PCR and ELISAs verified the reduction of IL-1ß and IL-18 in liver tissues. Mechanistic investigations indicated that PFDA inhibited caspase-1 activation, and decreased the mRNA levels of NLRP1, NLRP3, and NLRC4; thus, suggesting that inflammasome assemblies were suppressed. Further microarray data revealed that cIAP2 and its binding proteins, which are critical for regulating inflammasome assembly, were also repressed by PFDA. In addition, flow cytometry results revealed a significant inhibition of Th1 cell differentiation in the livers of PFDA-treated mice. CONCLUSIONS: The results of this study suggested that one of the main toxic effects of PFDA on livers was the inhibition of immune response.

XRCC4, which is inhibited by PFDA, regulates DNA damage repair and cell chemosensitivity.

The mechanism of environmental pollution promoting gastric cancer incidence and difficulty of treatment is not fully understood. In the present article, perfluorodecanoic acid (PFDA), a common persistent environmental pollutant, was used to treat the gastric cell lines and mice to test its genotoxicity. The γ-H2AX immunoblot and plasmid fragment PCR results showed that PFDA had a promotion effect on the DNA double-strand breaks (DSBs) in human and mouse cells. Subsequent results showed that PFDA significantly altered the sensitivity of cells to chemotherapy. Microarray data showed that the expressions of some important DNA repair genes were changed. Further investigation discovered that PFDA inhibition of DNA repair was mediated by X-ray repair cross complementing 4 (XRCC4). The cells deficient in XRCC4 generally exhibited reduced proliferation and premature aging in culture; however, our results indicated that PFDA induced p53 inhibition rescued cells from the apoptosis that was triggered by nonhomologous end-joining (NHEJ) inactivation, and overexpression of p53 expression in PFDA-treated cells enhanced their apoptosis. Finally, T-cell specific factor 4 was suggested by the results as an upstream regulator of XRCC4. This article revealed for the first time that perfluorinated chemicals affect chemotherapeutic sensitivity and the NHEJ pathway, and p53 reduction rescues cells from death.

HOX transcript antisense RNA is elevated in gastric carcinogenesis and regulated by the NF-κB pathway.

The expression pattern of HOX transcript antisense RNA (HOTAIR) in the progression of gastric cancer and the regulation of its expression are still unclear. In the current study, HOTAIR expressions in gastric tissues collected from patients with superficial gastritis, atrophic gastritis, atypical hyperplasia, and gastric cancer as well as normal controls was quantitatively examined. The results showed that the expression of HOTAIR was higher in gastric cancer than in normal tissues, but reached the highest level in atrophic gastritis, suggesting that HOTAIR may be involved in the molecular process of nonresolving inflammation. Then tumor necrosis factor-α-induced protein-8 like-2 (TIPE2), a known gene associated with nonresolving inflammation, was overexpressed and the results showed that the promotion in TIPE2 expression triggered HOTAIR reduction, this result was further verified by microarray analysis and TIPE2 knockout mice. Subsequently, the data obtained from HOTAIR knockdown experiment showed that it significantly enhanced colony forming capability and inhibited p27 expression in AGS cells. Furthermore, deletion constructs and luciferase-based activity assays indicated that the -475 to -443bp region of HOTAIR promoter contained a crucial regulatory element. Transcription factor prediction with software TRANSFAC revealed that nuclear factor-κB signaling protein p65 had a binding site in this region and might have roles in HOTAIR expression. The binding of phosphor-p65 to HOTAIR promoter was verified by chromatin immunoprecipitation, and succeeding experiment results demonstrated that p65 reduction by p65 small interfering RNA and TIPE2 overexpression also decreased HOTAIR expression. Conclusively, our results suggest that HOTAIR was associated with nonresolving inflammation, and its expression is regulated by p65.

[Clinical features and risk factors of pertussis in children].

OBJECTIVE: To study the clinical features and risk factors of pertussis in children. METHODS: A retrospective analysis was performed for the clinical data and laboratory markers for immune function of 253 hospitalized children with pertussis. A total of 314 hospitalized children with cough were used as the control group. Quantitative real-time PCR was used to detect Bordetella pertussis DNA. The clinical data of both groups were collected to analyze the risk factors for pertussis. RESULTS: A total of 23 typical clinical parameters were compared between the pertussis and control groups, and there were significant differences in only 10 clinical parameters between the two groups (P<0.01). As for the complications observed in the two groups, the pertussis group had a significantly lower incidence rate of myocarditis than the control group (P<0.05). The pertussis group had significantly lower levels of serum globulin and IgM than the control group (P<0.05). Compared with the control group, the pertussis group had a significantly higher proportion of children with a lack of diphtheria-pertussis-tetanus immunization or timely immunization and a contact history of suspected pertussis patients (P<0.05). A lack of vaccine immunization or timely immunization and a contact history of suspected pertussis patients were risk factors for pertussis (P<0.05). CONCLUSIONS: The clinical features are not typical in children with pertussis. Quantitative real-time PCR for detecting Bordetella pertussis DNA helps with the early diagnosis of atypical pertussis. Infants/toddlers should be immunized in time and be isolated from suspected pertussis patients to reduce the incidence of pertussis.

Perfluorodecanoic acid (PFDA) promotes gastric cell proliferation via sPLA2-IIA.

The association of perfluorodecanoicacid (PFDA) with tumor promotion and associated effects is not clear. Given that PDFA is mostly consumed with food and drinking water, we evaluated the effects of PFDA on a gastric cell line. When added to cell cultures, PFDA significantly increased growth rate and colony forming ability compared with control treatment. We found that suppression of cell senescence, but not apoptosis or autophagy was associated with PFDA-induced promotion of cell amount. To determine the molecular mechanism that was involved, DNA microarray assays was used to analyze changes in gene expression in response to PFDA treatment. Data analysis demonstrated that the vascular endothelial growth factor signaling pathway had the lowest p-value, with sPLA2-IIA (pla2g2a) exhibits the most altered expression pattern within the pathway. Moreover, sPLA2-IIA and its transcription factor TCF4, known as a direct target and a binding partner of Wnt/ß-catenin signaling in gastric cells respectively, were the third and second most varied genes globally. Cells transfected with expression plasmids pENTER-tcf4 and pENTER-pla2g2a show reduced cell proliferation by more than 60% and 30% respectively. Knockdown with sPLA2-IIA siRNA provided additional evidence that sPLA2-IIA was a mediator of PFDA-induced cell senescence suppression. The results suggest for the first time that PFDA induced suppression of cell senescence through inhibition of sPLA2-IIA protein expression and might increased the proliferative capacity of an existing tumor.

Alteration of stomach microbiota compositions in the progression of gastritis induces nitric oxide in gastric cell.

Atrophic gastritis is considered to be an antecedent to intestinal metaplasia and gastric cancer. A previous study identified that Helicobacter pylori was absent at the severe atrophic gastritis stage, and alterations in the gastric microbial composition resembled those in gastric cancer. To explore the role of the bacteria absence of H. pylori in gastric carcinogenesis, in the current study, we compared the microbiota of clinically collected H. pylori-free gastric fluids from 30 patients with non-atrophic gastritis (N) and 22 patients with severe atrophic gastritis (S). We estimated the bacterial loads in the N and S groups by colony counting in culture agar as well as by measuring the concentration of the extracted DNA. The results showed a significant increase in bacterial load in patients with atrophic gastritis in comparison to non-atrophic gastritis. Then, we analyzed the microbial communities of the gastric fluids from all 52 patients using high-throughput sequencing of 16S rRNA amplicons. The Chao 1, Shannon and Simpson diversity indexes demonstrated that the bacterial richness and diversity were not significantly different between the N and S groups. Moreover, principal component analysis illustrated that the microbiomes from the S group were more scattered. Microbiota composition analysis showed that the entire dataset was clustered into 27 phyla, 61 classes, 106 orders, 177 families, 292 genera and 121 species. At the genus level, only the abundance of Prevotella was significantly different between the N and S groups. Further analysis showed that all the higher taxonomic categories were significantly different between the N and S groups. To assess the effects of the metabolic products of Prevotella spp. on gastric cell physiology, we treated the human gastric epithelial cell line AGS with acetic acid and monitored nitric oxide (NO) production. The results showed that acetic acid at low concentrations (0.5 and 5 µM) significantly inhibited AGS cells to secrete NO compared to phosphate buffer saline-treated control cells. These results suggest that the microbiota in non-atrophic gastritis may influence gastric epithelial cell physiology.

Low-molecular-weight heparin protects kidney through an anti-apoptotic mechanism in a rat pre-eclamptic model.

OBJECTIVE: To investigate the effect of low-molecular-weight heparin (LMWH) for the treatment of pre-eclampsia (PE) and the underlying mechanism. STUDY DESIGN: PE was established in a rat model using l-NG-nitroarginine methyl ester (l-NAME). The effect of LMWH was examined by measuring various physiological parameters known to be associated with PE. RESULTS: Blood pressure, urinary protein, blood urea nitrogen and serum creatinine were higher in l-NAME-treated rats compared with control rats. In addition, the number of fetuses, and the weight of fetuses and placentas was lower in l-NAME-treated rats, indicating the establishment of PE conditions. Apoptosis was found in kidney tissue in l-NAME-treated rats. LMWH treatment restored the PE-associated disorders, and inhibited apoptosis in kidney tissue in these rats. CONCLUSIONS: LMWH can control PE conditions, protect renal function and improve fetal health. The mechanism of renal protection is likely to be related to the inhibitory effect of LMWH on apoptosis in kidney tissue. This study provides evidence that LMWH can potentially be used as a safe and effective anti-PE drug to improve PE conditions and protect renal function by inhibiting PE-induced apoptosis.

Corn peptides protect against thioacetamide-induced hepatic fibrosis in rats.

Certain bioactive peptides are reported to be able to alleviate hepatic fibrosis. Our previous work has confirmed the hepatoprotective effect of corn peptides (CPs) that are prepared from a high protein by-product, corn gluten meal, on acute liver injury in an animal model. However, the antifibrotic activity of CPs remained to be elucidated. In this study, the hepatoprotective effect of CPs on thioacetamide (TAA)-induced liver fibrosis was tested. Results showed that CPs (100 mg/kg body weight) significantly decreased the levels of alanine transaminase/aspartate transaminase, laminin, type IV collagen, and type III collagen in serum and increased the serum albumin levels and total antioxidant capacity. Additionally, with CP treatment (100 mg/kg body weight), a significant decrease was observed in the levels of malondialdehyde, nitric oxide, hydroxyproline, transforming growth factor ß1, and lactate dehydrogenase activity as well as the liver index, while the activity of superoxidedismutase was significantly increased in livers. The histological and morphological analysis showed that the hepatocyte structure in CP-treated rats was superior to that of TAA-injured rats, and inflammation and fibrosis were also ameliorated. Therefore, CPs can be used as an option for prevention and adjuvant therapy of liver fibrosis.

[Low molecular weight heparin inhibits cell apoptosis in the placenta of rats with preeclampsia-like symptoms].

OBJECTIVE: To investigate the anti-apoptotic effect of low molecular weight heparin (LMWH) in the placenta of rats with preeclampsia-like symptoms. METHODS: Thirty pregnant Wistar rats were randomized equally into 3 groups and received subcutaneous saline injection (control group), 200 mg/kg L-NAME injection to induce preeclampsia symptoms (PE group), or L-NAME with 40 µl/kg LMWH injections (LMWH group). The blood pressure, urine protein, the number of pups and the weight of the fetuses and placenta were measured, and the placental apoptotic index was measured by TUNEL assay. The expression of cleaved caspase-3, Bcl-2 and Bax in the placenta were examined by Western blotting. RESULTS: Compared with the control group, L-NAME injections caused significantly increased blood pressure and urine protein (P<0.05), which were significantly lowered by LWMH (P<0.05). The number and weight of normal pups were significantly lower in PE group than in the control group (P<0.05) and LMWH group (P<0.05); in LMWH group, the weight of pups was still lower than that of the control group (P<0.05) but the number of normal pups was comparable (P>0.05). The LMWH group showed a significantly lower placental apoptosis index than the PE group (P<0.05) with also significantly lower cleaved caspase-3 and Bax and higher Bcl-2 expressions (P<0.05). The apoptosis index and expressions of cleaved caspase-3, Bcl-2 and Bax protein were similar between LMWH group and normal pregnant group (P>0.05). CONCLUSION: LMWH can alleviate preeclampsia-like symptoms and decrease the apoptosis in the placenta of rats possibly by enhancing Bcl-2 and suppressing Bax expressions.

Multi-species comparison of the mechanism of biotransformation of MeO-BDEs to OH-BDEs in fish.

Polybrominated diphenyl ethers (PBDEs) and their methoxylated- (MeO-) and hydroxylated- (OH-) analogs are ubiquitously distributed in the environment worldwide. The OH-BDEs have greater potency than PBDEs and can be produced from the transformation of MeO-BDEs. The objectives of the current study were to (1) identify the enzyme(s) that catalyze biotransformation of 6-MeO-BDE-47 to 6-OH-BDE-47 in livers from rainbow trout, and (2) compare biotransformation of 6-MeO-BDE-47 to 6-OH-BDE-47 among rainbow trout, white sturgeon and goldfish. Cytochrome P450 1A (CYP1A) enzymes did not catalyze the biotransformation reaction. However, biotransformation was significantly inhibited by the CYP inhibitors clotrimazole and 1-benzylimidazole but not gestodene. Therefore, the reaction is likely catalyzed by CYP2 enzymes. When biotransformation was compared among species, concentrations of 6-OH-BDE-47 were significantly 3.4- and 9.1-fold greater in microsomes from rainbow trout compared to goldfish or white sturgeon, respectively. Concentrations of 6-OH-BDE-47 in microsomes from goldfish were non-significantly 2.7-fold greater than in sturgeon. The initial rate of biotransformation in microsomes from livers of rainbow trout was significantly 2.0- and 6.2-fold greater than the initial rate of biotransformation in microsomes from livers of goldfish or sturgeon, respectively, while the initial rate in goldfish was significantly 3.1-fold greater than in sturgeon. It is hypothesized that differences in CYP-mediated biotransformation of MeO-BDEs to OH-BDEs could influence concentrations of OH-BDEs in different species of fish.

Interconversion of hydroxylated and methoxylated polybrominated diphenyl ethers in Japanese medaka.

Polybrominated diphenyl ethers (PBDEs), hydroxylated (OH) and methoxylated (MeO), have been widely detected in aquatic environments. However, relationships among these structurally related compounds in exposed organisms are unclear. To elucidate biotransformation relationships among BDE-47, 6-OH-BDE-47, and 6-MeO-BDE-47, dietary accumulation, maternal transfer, and tissue distribution of these compounds and their transformation products were investigated in sexually mature Japanese medaka (Oryzias latipes). In addition, transformation of each compound was determined in vitro using liver microsomes of medaka. OH-PBDEs and MeO-PBDEs were not detected in fish exposed to BDE-47. However, significant concentrations of 6-OH-BDE-47 were detected in medaka or microsomes exposed to 6-MeO-BDE-47. Significant concentrations of 6-MeO-BDE-47 were also measured in fish exposed to 6-OH-BDE-47, but 6-MeO-BDE-47 was not detected in microsomes exposed to 6-OH-BDE-47. Similar patterns of transformation products were observed in medaka eggs from adult fish during exposure. This study presents direct in vivo evidence of biotransformation of 6-MeO-BDE-47 to 6-OH-BDE-47. In addition, this is the first study to demonstrate biotransformation of 6-OH-BDE-47 to 6-MeO-BDE-47. Demethylation of 6-MeO-BDE-47 was the primary transformation pathway leading to formation of 6-OH-BDE-47 in medaka, while the previously hypothesized formation of OH-PBDEs from synthetic BDE-47 did not occur. Biotransformation products formed in adult female medaka were transferred to eggs.

Polybrominated diphenyl ethers and their methoxylated metabolites in anchovy (Coilia sp.) from the Yangtze River Delta, China.

BACKGROUND, AIM, AND SCOPE: Polybrominated diphenyl ethers (PBDEs) and their metabolites are toxic to animals, and concentrations of the PBDEs metabolites can exceed those of the parent materials. But no information was available on concentrations of PBDEs metabolites in the lower Yangtze River in the region around Jiangsu Province of China, which is heavily urbanized and industrialized area. The aims of this study were to determine whether PBDEs and their methoxylated PBDEs (MeO-PBDEs) were accumulated in Coilia sp. in this area and to investigate the potential sources for these two kinds of brominated organic pollutants. MATERIALS AND METHODS: Samples of four species of anchovy were collected from eight sites in the lower Yangtze River, Taihu Lake, and Hongzehu Lake. Concentrations of 13 PBDEs congeners and eight methoxylated PBDEs were determined by use of organic solvent extraction, followed by gas chromatography and mass spectrometry. RESULTS AND DISCUSSION: The frequencies of detection for PBDEs and MeO-PBDEs were 92% and 53%, respectively. Concentrations of summation operatorPBDEs ranged from not detected (ND) to 77 ng/g lipids (ND-3.8 ng/g wet weight). Concentrations of summation operatorMeO-PBDEs in anchovy ranged from ND to 48 ng/g lipids (ND-8.2 ng/g wet weight). The PBDE concentrations in anchovy from the Yangtze River Delta were similar to or less than those reported for other species from other locations around the world, while the concentrations of MeO-PBDEs were comparable to or slightly less than those reported in other studies. This is the first report of MeO-PBDEs in biota of China. CONCLUSIONS: The results of this study as well as those of other studies suggest that PBDEs in anchovy are primarily of synthetic origin and released by human activities, while MeO-PBDEs in anchovy are primarily from nature as natural products from the sea instead of metabolism of PBDEs in anchovy.

Risk to humans of consuming metals in anchovy (Coilia sp.) from the Yangtze River Delta.

Concentrations of metals were determined in four species of anchovy (Coilia sp.) from the Yangtze River, Taihu Lake, and Hongze Lake in Jiangsu Province, China. Concentrations of Cr in anchovy fish muscle ranged from 2.6 × 10(-2) to 5.0 mg/kg ww, and Coilia nasus taihuensis in Jiaoshan, Taihu Lake contained the highest concentrations of Cr, which was almost 111-fold higher than the mean value at other locations. Concentrations of Pb ranged from 1.5 × 10(-2) to 1.3 × 10(-1) mg/kg ww. Comparisons of concentrations of lead (Pb) among the four species indicated that anadromous species contained higher concentrations of Pb than did freshwater species. However, concentrations of Pb in C. nasus from the Nanjing and Haimen locations in the Yangtze River were not significant higher than those of two freshwater species: C. nasus taihuensis from Taihu Lake and C. brachygnathus from Hongze Lake (Duncan's test, α = 0.05). While concentrations of Cd and Zn ranged from 7.0 × 10(-4) to 3.6 × 10(-3) mg/kg ww and 3.4 to 4.8 mg/kg ww, respectively, there were no significant differences in concentrations among the eight locations. The only concentration of the metals studied that exceeded the Chinese National Standard was Cr in Coilia from Jiaoshan, Taihu Lake, which was 2.5-fold higher than the standard. These results indicate that people who consume the genus Coilia are not at risk due to concentrations of metals, except Cr in C. nasus taihuensis from Jiaoshan in Taihu Lake. Concentrations of all of the metals studied except for Cr were similar to or less than those of metals in most other areas in the world.

Herbs for medicinal baths among the traditional Yao communities of China.

Medicinal baths are an important traditional way to prevent and cure common diseases among the traditional Yao communities of Jinping County, Yunnan Province, SW China. Approaches of anthropology, ethnobotany, and participatory rural appraisal (PRA) were used to investigate the herbs used for medicinal baths; and 110 medicinal plant species were found to be used by local people to treat a variety of diseases, such as rheumatic diseases, skin diseases, injuries from falls and gynecopathia. Of these 110 species, 6 (5%) had not been previously identified as having medicinal properties, while 87 (79%) were newly recorded for their use in medicinal baths. These new ethnobotanical and medicinal records are a rich source of further phytochemical, pharmacological, and clinical studies on folk herbs in SW China.