Sialic acid-based probiotic intervention in lactating mothers improves the neonatal gut microbiota and immune responses by regulating sialylated milk oligosaccharide synthesis via the gut-breast axis.

Human milk oligosaccharides (HMOs) are vital milk carbohydrates that help promote the microbiota-dependent growth and immunity of infants. Sialic acid (SA) is a crucial component of sialylated milk oligosaccharides (S-MOs); however, the effects of SA supplementation in lactating mothers on S-MO biosynthesis and their breastfed infants are unknown. Probiotic intervention during pregnancy or lactation demonstrates promise for modulating the milk glycobiome. Here, we evaluated whether SA and a probiotic (Pro) mixture could increase S-MO synthesis in lactating mothers and promote the microbiota development of their breastfed neonates. The results showed that SA+Pro intervention modulated the gut microbiota and 6'-SL contents in milk of maternal rats more than the SA intervention, which promoted Lactobacillus reuteri colonization in neonates and immune development. Deficient 6'-SL in the maternal rat milk of St6gal1 knockouts (St6gal1-/-) disturbed intestinal microbial structures in their offspring, thereby impeding immune tolerance development. SA+Pro intervention in lactating St6gal1± rats compromised the allergic responses of neonates by promoting 6'-SL synthesis and the neonatal gut microbiota. Our findings from human mammary epithelial cells (MCF-10A) indicated that the GPR41-PI3K-Akt-PPAR pathway helped regulate 6'-SL synthesis in mammary glands after SA+Pro intervention through the gut - breast axis. We further validated our findings using a human-cohort study, confirming that providing SA+Pro to lactating Chinese mothers increased S-MO contents in their breast milk and promoted gut Bifidobacterium spp. and Lactobacillus spp. colonization in infants, which may help enhance immune responses. Collectively, our findings may help alter the routine supplementation practices of lactating mothers to modulate milk HMOs and promote the development of early-life gut microbiota and immunity.

Alterations of milk oligosaccharides in mothers with gestational diabetes mellitus impede colonization of beneficial bacteria and development of RORγt+ Treg cell-mediated immune tolerance in neonates.

Gestational diabetes mellitus (GDM) is an increasing public health concern that significantly increases the risk of early childhood allergic diseases. Altered maternal milk glycobiome may strongly affect gut microbiota and enteric-specific Treg cell-mediated development of immune tolerance in GDM infants. In this study, we found that, compared with healthy Chinese mothers, mothers with GDM had significantly lower levels of total and specific human milk oligosaccharides (HMOs) in their colostrum that subsequently increased with extension of lactation. This alteration in HMO profiles significantly delayed colonization of Lactobacillus and Bifidobacterium spp. in their breast-fed infants, resulting in a distinct gut microbial structure and metabolome. Further experiments in GDM mouse models indicated that decreased contents of milk oligosaccharides, mainly 3'-sialyllactose (3'-SL), in GDM maternal mice reduced colonization of bacteria, such as L. reuteri and L. johnsonii, in the neonatal gut, which impeded development of RORγt+ regulatory T (Treg) cell-mediated immune tolerance. Treatment of GDM neonates with 3'-SL, Lactobacillus reuteri (L. reuteri) and L. johnsonii promoted the proliferation of enteric Treg cells and expression of transcription factor RORγt, which may have contributed to compromising ovalbumin (OVA)-induced allergic responses. In vitro experiments showed that 3'-SL, metabolites of L. johnsonii, and lysates of L. reuteri stimulated differentiation of mouse RORγt+ Treg cells through multiple regulatory effects on Toll-like receptor, MAPK, p53, and NOD-like receptor signaling pathways. This study provides new ideas for the development of gut microbiota and immune tolerance in GDM newborns.

Prediction of a miRNA-mRNA functional synergistic network for cervical squamous cell carcinoma.

Cervical squamous cell carcinoma (CSCC) accounts for a significant proportion of cervical cancer; thus, there is a need for novel and noninvasive diagnostic biomarkers for this malignancy. In this study, we performed integrated analysis of a dataset from the Gene Expression Omnibus database to identify differentially expressed genes (DEGs) and differentially expressed miRNAs (DEmiRNAs) between CSCC, cervical intraepithelial neoplasia (CIN) and healthy control subjects. We further established protein-protein interaction and DEmiRNA-target gene interaction networks, and performed functional annotation of the target genes of DEmiRNAs. In total, we identified 1375 DEGs and 19 DEmiRNAs in CIN versus normal control, and 2235 DEGs and 33 DEmiRNAs in CSCC versus CIN by integrated analysis. Our protein-protein interaction network indicates that the common DEGs, Cyclin B/cyclin-dependent kinase 1 (CDK1), CCND1, ESR1 and Aurora kinase A (AURKA), are the top four hub genes. P53 and prostate cancer were identified as significantly enriched signaling pathways of common DEGs and DEmiRNA targets, respectively. We validated that expression levels of three DEGs (TYMS, SASH1 and CDK1) and one DEmiRNA of hsa-miR-99a were altered in blood samples of patients with CSCC. In conclusion, a total of four DEGs (TYMS, SASH1, CDK1 and AURKA) and two DEmiRNAs (hsa-miR-21 and hsa-miR-99a) may be involved in the pathogenesis of CIN and the progression of CIN into CSCC. Of these, TYMS is predicted to be regulated by hsa-miR-99a and SASH1 to be regulated by hsa-miR-21.

Increased expression of human macrophage metalloelastase (MMP-12) is associated with the invasion of endometrial adenocarcinoma.

To evaluate the association between the expression of human macrophage metalloelastase (matrix metalloproteinase-12, MMP-12) with cancer invasion and differentiation of endometrial adenocarcinoma, specimens from endometrial adenocarcinoma (n=61) of diverse stages and histologic types were collected from patients having undergone hysterectomy, and specimens from normal endometrium (n=38) were obtained from patients with benign diseases. The expression of MMP-12 was analyzed immunohistochemically, by Western blot, and RT-PCR. The positive rate of MMP-12 was significantly increased in endometrial adenocarcinoma (81.97%) as compared with that in normal endometrium (13.16%). The results showed that expression of MMP-12 correlated with stage (p=0.022) and grade (p=0.018) of endometrial cancer. MMP-12 immunoreactive proteins were found mainly on the glandular epithelial cells of endometrial adenocarcinoma. The macrophage infiltration detected by CD68 immunohistochemical staining in endometrial adenocarcinoma was also higher than that in normal endometrium. In this study, we show that in addition to macrophages, endometrial adenocarcinoma cells are able to express MMP-12. Increased MMP-12 expression tended to be associated with the extent of adenocarcinoma invasion accompanied by marked macrophage infiltration. Our results suggest that MMP-12 is an important oncogene in high-stage and high-grade endometrial adenocarcinoma.

Synchronous leiomyosarcoma and fibroma in a single ovary: A case report and review of the literature.

Primary ovarian leiomyosarcoma (POLMS) is a rare disease. To the best of our knowledge, only 72 cases, including the present case, have been reported in the English literature, while synchronous POLMS and fibroma in a single ovary have not previously been reported at all. In the present study, a 46-year-old premenopausal woman was diagnosed with a mass in the left ovary in 2005. A total of 5 years after the diagnosis of this mass, the patient was admitted to hospital exhibiting lower abdominal pain, and two masses were observed in the left ovary. An exploratory laparoscopy was performed. Frozen section analysis led to a diagnosis of fibroma. Furthermore, the observed second mass was hypothesized to be a malignant form of the original fibroma. A hysterectomy and bilateral salpingo-oophorectomy were performed. Pathological reports following surgery revealed concurrent stage Ic POLMS and fibroma in the left ovary. A total of 13 months after the initial surgery, recurrent leiomyosarcoma was detected. Although the patient underwent multiple cytoreductive surgeries and chemotherapy cycles, as well as interstitial brachytherapy and conventional therapy, a poor state of health ensued. Due to the rarity of POLMS, particularly in combination with ovarian fibroma, the current report presents a detailed overview of the literature and discusses a number of histogenetic and clinical issues.

[Transfection and expression of hRI gene on human umbilical blood stem cells and gene therapy for mouse melanoma].

In order to explore the transfection and expression of hRI gene on human umbilical blood stem cells, and observe it's effect on the tumor growth. After enriching human umbilical cord blood CD34+ cells with a high-gradient magnetic cell sorting system (MACS), transfected them with supernatant of retrovirus containing human Ribonuclease inhibitor (hRI) cDNA. Hematopoietic progenitor clonogenic assay and PCR were used to evaluate transfection efficiency, and Western-blot and immune fluorescence were used to evaluate the expression quantity of hRI gene after transfection. Observe the effect of RI on the growth of melonoma in B16C57BL mice. The results showed that human umbilical blood CD34+ cells were highly purified by MACS, which made the purity of human umbilical blood CD34+ cells average 96.15%. hRI can be transfected on umbilical blood CD34+ cells, and the transfection efficiency was 35%. The positive expression of hRI gene on transfected CD34+ cells is identified by Western-blot and immune fluorescence assay. Mice injected with transfected CD34+ cells show a significant restraint of the tumor growth, a lower efficiency of tumor formation, a lower weight of the tumor and a longer incubation period of tumor formation with respect to the control groups. The results demonstrated the capacity of RI to inhibited the tumor growth by blocking the vasculature in tumor.