Saponin-Induced Shifts in the Rumen Microbiome and Metabolome of Young Cattle.

The aim of this study was to explore the effects of saponins on the rumen microbiota and the ruminal metabolome. Alfalfa hay (AH) and soybean hulls (SH) were used as fiber sources for the control diets. The AH and SH diets were supplemented with tea saponins resulting in two additional diets named AHS and SHS, respectively. These 4 diets were fed to 24 young male Holstein cattle (n = 6 per diet). After 28 days of feeding, the rumen fluid from these cattle was collected using an oral stomach tube. Illumina MiSeq sequencing and ultrahigh-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS) were used to investigate the changes in the ruminal microbes and their metabolites. The relative abundance of Prevotellaceae_YAB2003 increased, while Ruminococcaceae_NK4A214 and Lachnospiraceae_NK3A20 decreased in SHS and AHS compared to SH and AHS, respectively. Feeding SHS resulted in higher ruminal concentrations of squalene, lanosterol, 3-phenylpropanoic acid, and citrulline compared to SH. The different microbial genes predicted by Tax4Fun were involved in amino sugar and nucleotide sugar metabolism. The pathways of arginine and proline metabolism, purine metabolism, and pyrimidine metabolism were enriched by different metabolites. Moreover, in the SH group, a positive correlation was observed between Prevotella_1 (Bacteroidetes), Prevotellaceae_YAB2003 (Bacteroidetes), and Christensenellaceae_R.7 (Firmicutes), and the metabolites, including citrulline, lanosterol, and squalene. The increased abundances of Prevotella_1, Ruminococcaceae_UCG.002, and Prevotellaceae_YAB2003 might result in increased fiber digestion and nutrient utilization but nutrient digestion was not measured in the current study. In summary, saponins have the ability to modulate the ruminal microbial community and ruminal metabolites and thus affect the rumen environment. However, the response seems to be dependent on the composition of the basal diet. This study provides a comprehensive overview of the microbial and biochemical changes in the rumen of cattle fed saponins.

Expression of MIER3 in colorectal cancer and bioinformatic analysis of MIER3- interacting proteins / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore role of MIER3 gene in the development and progression of human colorectal carcinoma (CRC) and analyze the proteins that interact with MIER3 using bioinformatic techniques.</p><p><b>METHODS</b>MIER3 mRNA and protein expressions were detected in 8 CRC biopsy samples and paired adjacent tissues using real-time PCR and Western blotting. A recombinant eukaryotic expression vector pcDNA3-MIER3 was constructed and its effect on the proliferation and invasion of CRC cells were tested using CCK8 assay and Transwell migration assay. Bioinformatic methods were used to predict and analyze MIER3-interacting proteins.</p><p><b>RESULTS</b>MIER3 was obviously down-regulated in the 8 CRC tissues as compared with the paired adjacent tissues. In human CRC cell line DLD1, MIER3 overexpression induced by transfection of the cells with pcDNA3-MIER3 significantly inhibited the cell proliferation and suppressed cell invasiveness in vitro. Bioinformatics analyses indicated that NAT9 was a potential MIER3-interacting protein and MIER3 was probably associated with tumor susceptibility.</p><p><b>CONCLUSION</b>MIER3, which is obviously down-regulated in CRC tissues, is closely associated with the proliferation and invasion of CRC, and NAT9 protein is a probable MIER3-interacting protein.</p>

Preparation and Application of Porous Layer Open Tubular Capillary Columns with Narrow Bore in Liquid Chromatography / 分析化学

In recent years, micro/nano-scaled separation technique has attracted increasing attention due to its inherent advantages. The porous layer open tubular ( PLOT) capillary column is an important microcolumn form. Comparied to wide-bore PLOT capillary columns ( inner diameter >25 μm ) , the PLOT capillary columns with a narrow inner diameter yield higher separation efficiency and lower reagent consumption. In this paper, the preparation methods for porous layer open tubular capillary columns with narrow inner diameter, less than or equal to 25 μm, are reviewed. Detection techniques combined with mass spectrometry and their applications in liquid chromatography are also disussed.

Construction of a eukaryotic expression vector of TM4SF1 and its effect on migration and invasion of colorectal cancer cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a eukaryotic expression vector of transmembrane-4-L-six-family-1 (TM4SF1) gene and study its effect on the migration and invasion of colorectal cancer cells.</p><p><b>METHODS</b>A pair of specific primers of TM4SF1 gene (GenBank: BC034145.1) was used to acquire the open reading frame of TM4SF1 by RT-PCR. The amplified sequence was ligated to a PEZ-M29 vector, which, after identification, was transiently transfected in colorectal cancer cell lines HCT116 and DLD1. Western blotting and immunocytochemistry were used to analyze the transfection efficiency, and scratch and Transwell tests were performed to analyze the changes in the migration and invasion of HCT116 and DLD1 cells after transfection.</p><p><b>RESULTS</b>Cell scratch and Transwell assays revealed that transfection with the recombinant plasmid, PEZ-M29/TM4SF1, caused up-regulated expression of TM4SF1 and promoted the migration and invasion of HCT116 and DLD1 cells.</p><p><b>CONCLUSION</b>Our results demonstrated that TM4SF1 is closely related to the invasion and metastasis of colorectal cancer cells in vitro.</p>

[Microbial community structure analysis of unexploited oil and gas fields by PCR-DGGE].

Microbial communities of different depths (30, 60, 100, 150, 200cm) from the unexploited oilfield, gas field and control area were studied by PCR-DGGE and sequencing methods. The objectives of this study were to understand the microbial distribution in the regions of unexploited oil and gas fields, and to investigate the potential microbial indicators of oil and gas resources. The results showed that the Dice coefficients between different depths were very low (26-69.9). The microbial communities in the soil of 150 cm and 200 cm depth had greater richness (S > or = 19), diversity (H > or = 2.69) and evenness (E > or = 0. 90). The results of sequencing demonstrated that the bands from oilfield were mainly grouped into alpha-Proteobacteria, gamma-Proteobacteria, Actinobacteria, Acidobacteria with the predominance of gamma-Proteobacteria (75%). Most of the bands were related to oil-associated and hydrocarbon degrading bacteria, such as Methylophaga and Alcanivorax. While the gas field had alpha, beta, gamma, delta-Proteobacteria and Bacteroidetes, and gamma-Proteobacteria accounted for only 24%. More strains showed relativity to methanotrophs, such as Methylocystaceae. Thus, 150 cm and 200 cm were more suitable as the oil-gas exploration sampling depth. Methylocystaceae may act as potential indicators for gas resources, Methylophaga and Alcanivorax for oil.

Effect of calming the liver and suppressing the hyperactive YANG drugs on lymphocyte protein in hypertension patients with hyperactivity of liver-YANG / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of calming the liver and suppressing the hyperactive YANG drugs on the lymphocyte protein and clinical efficacy in the hypertension patients with hyperactivity of liver-YANG, and to identify the therapy.@*METHODS@#Twenty-six hypertension patients with hyperactivity of liver-YANG were treated by calming the liver and suppressing the hyperactive YANG drugs for 2 courses. Symptoms of Chinese medicine and blood pressure were observed, and the separated lymphocyte total protein of normal and hypertensions before and after the treatment were examined by the solid-state pH gradient 2-dimensional gel electrophoresis. The differences of the protein expression were analyzed by ImageMaster 2DE analysis software with two-way patterns.@*RESULTS@#The total efficiency rate of calming the liver and suppressing the hyperactive YANG drugs was 88.5%, and the drugs could significantly relieve the symptoms, such as headache, dizziness, dry mouth, irritability, etc. Calming the liver and suppressing the hyperactive YANG drugs could also remarkably reduce the blood pressure,with significant different between pre-treatment and post-treatment (P<0.05). The average spots of lymphocyte gel proteins in the normal and the hypertension patients with syndrome of hyperactivity of liver-YANG before and after the treatment were 527+/-41,559+/-62, and 543+/-59, respectively. Compared with normal people, the expression of 15 proteins was down-regulated, and 10 up-regulated in the hypertension patients with syndrome of hyperactivity of liver-YANG. Compared with the pre-treatment, the expression of 12 proteins was increased in the 15 down-regulated proteins, and 6 decreased in the 10 up-regulated proteins after the treatment in the hypertension patients with syndrome of the hyperactivity of liver-YANG.@*CONCLUSION@#Calming the liver and suppressing the hyperactive yang drugs may mildly depress the blood pressure and improve the symptoms of Chinese medicine. The effect of drugs in treating hypertension may probably be associated with regulating the expression of some proteins in lymphocytes.