Fecal microbiota transplantation improves metabolism and gut microbiome composition in db/db mice.

Fecal microbiota transplantation (FMT) has become an effective strategy to treat metabolic diseases, including type 2 diabetes mellitus (T2DM). We previously reported that the intestinal microbiome had significant difference between individuals with normal glucose tolerance and T2DM in Chinese Kazak ethnic group. In this study, we investigated the effects of transplanted fecal bacteria from Kazaks with normal glucose tolerance (KNGT) in db/db mice. The mice were treated with 0.2 mL of fecal bacteria solution from KNGT daily for 10 weeks. We showed that the fecal bacteria from KNGT successfully colonized in the intestinal tract of db/db mice detected on day 14. In the FMT-treated db/db mice, the levels of fasting blood glucose, postprandial glucose, total cholesterol, triglyceride, and low-density lipoprotein-cholesterol were significantly downregulated, whereas high-density lipoprotein-cholesterol levels were upregulated. In the FMT-treated db/db mice, Desulfovibrio and Clostridium coccoides levels in gut were significantly decreased, but the fecal levels of Akkermansia muciniphila and colon histone deacetylase-3 (HDAC3) protein expression were increased. At 8 weeks, both intestinal target bacteria and HDAC3 were correlated with glycolipid levels; Akkermansia muciniphila level was positively correlated with HDAC3 protein expression (r = +0.620, P = 0.037). Our results suggest that fecal bacteria from KNGT could potentially be used to treat diabetic patients.

Characterization of a novel Xenopus SH3 domain binding protein 5 like (xSH3BP5L) gene.

SH3 domain binding protein 5 like (xSH3BP5L) gene encodes a protein that is a new found member of SH3 domain binding protein family which has been implicated at multiple levels of biological functions. Here, we have characterized Xenopus SH3 domain binding protein 5 like (xSH3BP5L) gene in the development of Xenopus laevis. Transcripts of xSH3BP5L were detected at all stages of development and in numerous adult tissues. Whole-mount in situ hybridization demonstrated that xSH3BP5L is expressed at the animal pole from stage-2 onward. Interestingly, translational inhibition of xSH3BP5L using antisense morpholino oligonucleotides (MOs) and overexpression of xSH3BP5L in Xenopus embryos resulted in failed or delayed blastopore closure. Taken together, these data suggested that xSH3BP5L is required for normal embryogenesis of blastopore closure in X. laevis.

[Sequencing the Rhesus boxes and determining the homozygosity of RHD gene in Han Chinese with RhD negative].

OBJECTIVE: To analyze the sequences of Rhesus boxes of RhD gene, and explore the genetic mechanism of RhD negative phenotype in Chinese Han population. Meanwhile the PCR product of Rhesus boxes is analyzed for determining RHD gene homozygosity. METHODS: DNA of 74 RhD negative samples were firstly analyzed with multiplex PCR-sequence specific primer(SSP). The further analysis was given to Rhesus boxes specific sequencing and RHD gene homozygosity determined by PCR-restriction fragment length polymorphism(RFLP) analysis to Rhesus boxes. RESULTS: In DNA samples of 74 RhD negative individuals, 46 samples(62%) showed the absence and homozygous negative of RHD gene; 22 samples(30%) all showed the existence of RHD specific exons, of which 19 were RHD gene heterozygous and 3 were homozygous; regardless of PCR-RFLP analysis showing no RHD specific exons, but further analysis of RHD specific PCR revealed one RHD gene, at least RHD gene exon 1 and 10 existing in 5 DNA samples(7%); 1 sample(1%) was lacking RHD exon 6 although the multiplex PCR showed the RHD gene to be positive. Analyzing the hybrid Rhesus box of 27 RhD negative samples revealed the Han Chinese population to have the same DNA sequence of hybrid Rhesus box as Caucasians. CONCLUSION: The RHD gene deletion is the main molecular mechanism of causing RhD negative formed in Han Chinese population, who have had the RHD gene deletion taken place within the defined breakpoint region as Caucasians.

Expression of Pkd2l2 in testis is implicated in spermatogenesis.

Pkd2l2 is a novel member of the polycystic kidney disease (PKD) gene family in mammals. Prominently expressed in testis, this gene is still poorly understood. In this study, reverse transcription polymerase chain reaction (RT-PCR) results showed a time-dependent expression pattern of Pkd2l2 in postnatal mouse testis. Immunohistochemical analysis revealed that Pkd2l2 encoded a protein, polycystin-L2, which was predominantly detectable in the plasma membrane of spermatocytes and round spermatids, as well as in the head and tail of elongating spermatids within seminiferous tubules in mouse testis tissue sections of postnatal day 14 and adult mice. A green fluorescent fusion protein of Pkd2l2 resided in the plasma membrane of HEK 293 and MDCK cells, suggesting that it functions as a plasma membrane protein. Overexpression of Pkd2l2 increased the intracellular calcium concentration of MDCK cells, as detected by flow cytometry. Collectively, these data indicated that Pkd2l2 may be involved in the mid-late stage of spermatogenesis through modulation of the intracellular calcium concentration.

TC1 (C8orf4) is involved in ERK1/2 pathway-regulated G(1)- to S-phase transition.

Although previous studies have implicated a role for TC1 (C8orf4) in cancer cell proliferation, the molecular mechanism of its action is still largely unclear. In this study, we showed, for the first time, that the mRNA levels of TC1 were upregulated by mitogens (FBS/thrombin) and at least partially, through the ERK1/2 signaling pathway. Interestingly, the over-expression of TC1 promoted the G(1)- to S-phase transition of the cell cycle, which was delayed by the deficiency of ERK1/2 signaling in fibroblast cells. Furthermore, the luciferase reporter assay indicated that the over-expression of TC1 significantly increased Cyclin D1 promoter-driven luciferase activity. Taken together, our findings revealed that TC1 was involved in the mitogen-activated ERK1/2 signaling pathway and positively regulated G(1)- to S-phase transition of the cell cycle. Our results may provide a novel mechanism of the role of TC1 in the regulation of cell proliferation.