Autoimmunity to urothelial antigen causes bladder inflammation, pelvic pain, and voiding dysfunction: a novel animal model for Hunner-type interstitial cystitis.

Recent evidence revealed that Hunner-type interstitial cystitis (HIC) is a robust inflammatory disease potentially associated with enhanced immune responses and histologically characterized by epithelial denudation and lymphoplasmacytic infiltration with frequent clonal expansion of infiltrating B cells. To date, few animal models that reproduce the histological and clinical correlates of HIC have yet been established. In the present study, we aimed to develop a novel animal model for HIC via autoimmunity to the bladder urothelium using the transgenic mouse model (URO-OVA) that expresses the membrane form of the model antigen ovalbumin (OVA) as a self-antigen on the bladder urothelium. OVA-specific lymphocytes (splenocytes) were generated by immunization of C57BL/6 mice with OVA protein and injected intravenously into URO-OVA mice. The splenocytes from OVA-immunized C57BL/6 mice showed increased interferon (IFN)-γ production in response to OVA stimulation in vitro. URO-OVA mice adoptively transferred with OVA-primed splenocytes developed cystitis exhibiting histological chronic inflammatory changes such as remarkable mononuclear cell infiltration predominantly composed of T and B lymphocytes, increased vascularity, and mucosal hyperemia in the bladder at days 7-28 with a peak at day 21 tested. No systemic inflammation was found in cystitis-induced URO-OVA mice, nor was any inflammation found in wild-type C57BL/6 mice adoptively transferred with OVA-primed splenocytes. Along with bladder inflammation, URO-OVA mice demonstrated significantly increased pelvic nociceptive responses, voiding dysfunction, and upregulated mRNA expression levels for IFN-γ, tumor necrosis factor-α (TNF-α), and substance P precursor in the bladder. This model reproduces the histological and clinical features of human HIC, providing a novel model for HIC research.

[A preliminary study of the difference in composition of intestinal bifidobacteria between healthy infants and infants with allergic diseases].

OBJECTIVE: To investigate the association between intestinal bifidobacteria and allergic diseases in infants by comparing the composition of intestinal bifidobacteria between healthy infants and infants with allergic diseases. METHODS: A total of 48 infants were enrolled, and fecal samples were collected on days 0, 2, 7, and 15 and at months 1, 6, and 12 after birth. Among these infants, 22 who experienced allergic diseases before the age of 1 year were enrolled as allergic group and 26 healthy infants were enrolled as healthy group. Quantitative real-time PCR was used for the qualitative and quantitative analyses of Bifidobacterium and 8 species of bifidobacteria in fecal samples. RESULTS: There was a difference in the composition of intestinal bifidobacteria between the two groups within 1 month after birth: the healthy group showed a reduction in bifidobacteria on day 2, while this feature was not observed in the allergic group. Compared with the healthy group, the allergic group had a significantly lower detection count of Bifidobacterium at month 1 (P<0.05) and a significantly lower detection rate of B.breve on day 15 (P<0.05), with delayed colonization of B.infantis. CONCLUSIONS: Intestinal bifidobacteria and their composition within 1 month after birth may be associated with the development of allergic diseases, and this period of time may be a critical period for the prevention and treatment of allergic diseases in infants.

[A preliminary analysis of changes in composition of intestinal microbiota during infancy using polymerase chain reaction-denaturing gradient gel electrophoresis].

OBJECTIVE: To investigate the composition of bacteria in the stools of infants and the colonization of intestinal microbiota during infancy. METHODS: Fresh stools were collected from 15 healthy infants at 0, 2, 4, 7, 10, 14, and 28 days and 3, 6, and 12 months after birth. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the composition of intestinal microbiota, perform sequencing of dominant bacteria, and to analyze the changes in the composition of intestinal microbiota during infancy. RESULTS: DGGE fingerprint showed that the composition of intestinal microbiota during infancy changed significantly over time after birth. The cloning and sequencing results indicated that Proteobacteria colonized the earliest, mainly the obligate aerobes Enterobacter and Pseudomonas, followed by the obligate anaerobes (Clostridium hathewayi and Veillonella parvula) and the facultative anaerobe Clostridium ramosum in Firmicutes, and Verrucomicrobia. Actinobacteria colonized the latest, mainly Bifidobacterium, and gradually became dominant bacteria. CONCLUSIONS: During infancy, obligate aerobes colonize the intestinal tract the earliest, followed by obligate anaerobes and facultative anaerobes. Proteobacteria colonizes the earliest, followed by Firmicutes and Verrucomicrobia, and Actinobacteria, mainly Bifidobacterium, colonizes the latest.

[Analyzing Colonization of Bifidobacteria in Infants with Real-time Fluorescent Quantitative PCR].

OBJECTIVES: In order to know how intestinal Bifidobacteria community could be built in the infants and whether the environmental factors could affect them, the present study was conducted to characterizethe species composition and trace the quantitative changes of intestinal Bifidobacteria of the infants in their early stages with non-culture dependent molecular method. The possible association of Bifidobacteria community of the infants with their health was also discussed. METHODS: Total 16 of full-term newborn infants born between March and April 2013 were recruited for the present study. Fecal samples were collected from them at 1 day, 2 days, 4 days, 7 days, 10 days, 14 days, 28 days, 3 months, 6 months and 1 year after birth. Real-time fluorescent quantitative PCR with genus and species specific premiers was used to detect Bifidobacteria and 8 predominate species in human intestine qualitatively and quantitatively present in these collected fecal samples. RESULTS: Total 136 fecal sample were collected and Bifidobacteria were detected from 93.4% (127/136) of them with the concentration of 1.0×10 5 to 1.0×10 11 CFU/g. Bifidobacteria were found in 83.3% of the fecal samples collected from the first day after birth with more than about 10 5 CFU/g. However, Bifidobacteria were detected relative low until 14 days and were taxonomically belonged only to one or two species. Bifidobacteria were found in almost 100% of the fecal samples collected after birth 28 days with more than 108 CFU/g, and the detected species of Bifidobacteria was increased to 3 species after 28 days to 6 months. All of the fecal samples collected from one year had more than 3 species of Bifidobacteria with high cell counts. Among the detected Bifidobacteria were B.breve 92.1%, B.infantis 66.1%, B. catenulatum 59.8%, B. bifidum 25.2%, B. longum 24.4%, B.dentium 13.4%, B.angulatum 5.5% and B.adolescentis 1.6%, respectively. CONCLUSIONS: The detected Bifidobacteria greatly varied qualitatively and quantitatively after birth to one year which could be considered as the important and sensitive period for Bifidobacteria to colonize and built its communityin the infants. Different from previous studies, the colonization of Bifidobacteria in the tested infants was found delayed and the composition and diversity of Bifidobacteria species was different from other studies. These might result from different deliveryway, feeding pattern and other environmental factors related to the tested infants.

[Colonization and development of intestinal bifidobacteria in early infancy].

OBJECTIVE: To study the characteristics of the colonization of 8 species of bifidobacteria by systematically profiling fecal bifidobacterial community in the early life of infants. METHODS: Fresh fecal samples including meconium samples were collected for culture and isolation of fecal bifidobacteria from 16 cases of full-term newborn infants born between March and April 2013 at their life of 2, 4, 7, 10, 14, 28, and 90 days. The isolated fecal bifidobacteria were taxonomically identified to genus and 8 species with PCR analysis. RESULTS: One hundred and fifty-two predominant bifidobacteria strains were detected in the fecal samples, the detection rate of B. breve (22.4%) were the highest. Bifidobacteria were found in the feces of 8% infants 4 days after birth. The colonization rates increased to 54% and 60% at 28 days and 3 months respectively, significantly exceeding the colonization rate at 4 days after birth (P<0.05). Adult-type bifidobacteria B. catenulatum were found in the infants 10 days after birth, and infant-type bifidobacteria B. infantis were found at 14 days after birth, but infant-type bifidobacteria B. infantis were detected at a high level until 3 months after birth. The most tested infants had 2 species or less of bifidobacteria. CONCLUSIONS: Intestinal bifidobacteria in infants might have less diversity in early infancy. Infant-type bifidobacteria appear late, while adult-type bifidobacteria colonize earlier.

Programming of growth, insulin resistance and vascular dysfunction in offspring of late gestation diabetic rats.

ODM (offspring of diabetic mothers) have an increased risk of developing metabolic and cardiovascular dysfunction; however, few studies have focused on the susceptibility to disease in offspring of mothers developing diabetes during pregnancy. We developed an animal model of late gestation diabetic pregnancy and characterized metabolic and vascular function in the offspring. Diabetes was induced by streptozotocin (50 mg/kg of body weight, intraperitoneally) in pregnant rats on gestational day 13 and was partially controlled by twice-daily injections of insulin. At 2 months of age, ODM had slightly better glucose tolerance than controls (P<0.05); however, by 6 months of age this trend had reversed. A euglycaemic-hyperinsulinamic clamp revealed insulin resistance in male ODM (P<0.05). In 6-8-month-old female ODM, aortas had significantly enhanced contractility in response to KCl, ET-1 (endothelin-1) and NA (noradrenaline). No differences in responses to ET-1 and NA were apparent with co-administration of L-NNA (NG-nitro-L-arginine). Relaxation in response to ACh (acetylcholine), but not SNP (sodium nitroprusside), was significantly impaired in female ODM. In contrast, males had no between-group differences in response to vasoconstrictors, whereas relaxation to SNP and ACh was greater in ODM compared with control animals. Thus the development of diabetes during pregnancy programmes gender-specific insulin resistance and vascular dysfunction in adult offspring.

Polyunsaturated fatty acid composition and childhood adversity: Independent correlates of depressive symptom persistence.

Childhood experiences, personality, and polyunsaturated essential fatty acid (PUFA) composition have all been shown to affect the likelihood of depressive symptoms. Few studies have addressed relationships between these factors in their influence on the occurrence or course of depressive symptoms. The following analysis was designed to do so. Subjects, 15-20 years old, had either begun antidepressant treatment within the preceding month (n = 88), or had never taken psychiatric medications (n = 92). Baseline assessments included a structured diagnostic interview, the self-completed Multiphasic Personality Questionnaire, and a determination of plasma PUFA phospholipid composition. Depressive symptom levels were assessed at baseline and again at 4, 8 and 12 months. Omega-3 composition and general childhood trauma scores were unrelated to each other but both correlated, in predicted directions, with negative emotionality. Low omega-3 composition and history of childhood trauma were associated with persistence of depressive symptoms during follow-up, largely through their effects on negative emotionality. Negative emotionality appears to comprise a final common pathway to depressive disorder through which the diverse risk factors of childhood adversity and low omega-3 composition are expressed.