[Expression of miR-140-5p and prediction of its target gene in human mesenchymal stem cells during adipogenic differentiation].

OBJECTIVE: To screen the differentially expressed miRNAs and their target genes in adipogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs) to better understand the mechanism for regulating the balance between osteoblast and adipocyte differentiation. METHODS: Cultured hMSCs were induced for adipogenic differentiation, and at 0, 7, 14, and 21 days of induction, the cells were examined for miRNA and mRNA expression profiles using miRNA chip and transcriptome sequencing (RNA-seq) techniques. Correlation analysis was carried out for the miRNAs and mRNAs of potential interest. The databases including TargetScan, PicTar and miRanda were used to predict the target genes of the differentially expressed miRNA. RESULTS: The expression of miR-140-5p was down-regulated and leukemia inhibitory factor receptor (LIFR) expression increased progressively during adipogenic differentiation of hMSCs, showing a negative correlation between them. Target gene prediction using the 3 databases identified LIFR as the target gene of miR-140-5p. CONCLUSION: miRNA-140-5p may play an important role by regulating its target gene LIFR during adipogenic differentiation of hMSCs.

[Cloning of IPO8 promoter and analysis of its transcription activity].

OBJECTIVE: To clone 5' untranslated region of human IPO8 gene and determine its transcription activity. METHODS: We used 5' rapid amplification of cDNA ends (RACE) analysis to identify the IPO8 transcription start site (TSS), and amplified series truncated 5' UTR fragment containing transcription start site. The PCR productions were inserted into luciferase report vector pGL3- Basic. After confirmation by restriction enzyme digestion, the recombinant plasmids were cotransfected into Saos-2 cells with plasmid pRL-TK. The luciferase activities were measured by dual luciferase reporter system. RESULTS: The IPO8 gene transcription start site was established. The electrophoresis analysis of restriction enzyme digestion and DNA sequencing verified the fragments were successfully amplified and inserted into pGL3-Basic. After the recombinant plasmids transfected, the highexpressions of luciferase were detected in Saos-2 cells. CONCLUSION: The recombinant vector containing IPO8 promoter is constructed successfully, which provides a foundation for determining expressional regulation of IPO8 in the further study.

[Analysis on published papers and its citation in Chinese Journal of Schistosomiasis Control, 2008-2010].

OBJECTIVE: To study the published papers and its citation of Chinese Journal of Schistosomiasis Control, so as to provide evidence for the improvement of its quality. METHODS: The published papers in Chinese Journal of Schistosomiasis Control from 2008 to 2010 were searched, biblimetrics analysis was employed, and the number of published papers, the proportion of fund articles, the cooperation rate of scientific research, the distribution of authors, the citations and their types, the half-life periods and the Price's index were analysed. RESULTS: From 2008 to 2010, the number of papers published were 217, 203, 210, respectively, and the proportions of fund articles were 32.72%, 38.92% and 49.52%, respectively. The cooperation rates of scientific research were 87.56%, 95.07%, 94.76%, respectively. The average amounts of citation were 5.49, 10.14, 13.33 per paper, respectively, and the citations were mainly from books and journals. The Price's indexes were 47.23%, 50.12% and 51.48%, respectively. CONCLUSIONS: The quality and academic level of papers published in Chinese Journal of Schistosomiasis Control is improving year by year, the journal can satisfy its authors and readers with the latest information of scientific research in schistosomiasis and parasitic diseases study and control. But it requires to enlarge the author group and to increase the amount of citation to its further development.

Decreased ENaC expression compensates the increased NCC activity following inactivation of the kidney-specific isoform of WNK1 and prevents hypertension.

Mutations in WNK1 and WNK4 lead to familial hyperkalemic hypertension (FHHt). Because FHHt associates net positive Na(+) balance together with K(+) and H(+) renal retention, the identification of WNK1 and WNK4 led to a new paradigm to explain how aldosterone can promote either Na(+) reabsorption or K(+) secretion in a hypovolemic or hyperkalemic state, respectively. WNK1 gives rise to L-WNK1, an ubiquitous kinase, and KS-WNK1, a kinase-defective isoform expressed in the distal convoluted tubule. By inactivating KS-WNK1 in mice, we show here that this isoform is an important regulator of sodium transport. KS-WNK1(-/-) mice display an increased activity of the Na-Cl cotransporter NCC, expressed specifically in the distal convoluted tubule, where it participates in the fine tuning of sodium reabsorption. Moreover, the expression of the ROMK and BKCa potassium channels was modified in KS-WNK1(-/-) mice, indicating that KS-WNK1 is also a regulator of potassium transport in the distal nephron. Finally, we provide an alternative model for FHHt. Previous studies suggested that the activation of NCC plays a central role in the development of hypertension and hyperkalemia. Even though the increase in NCC activity in KS-WNK1(-/-) mice was less pronounced than in mice overexpressing a mutant form of WNK4, our study suggests that the activation of Na-Cl cotransporter is not sufficient by itself to induce a hyperkalemic hypertension and that the deregulation of other channels, such as the Epithelial Na(+) channel (ENaC), is probably required.

Regulation of WNK1 expression by miR-192 and aldosterone.

WNK1 and WNK4 encode two members of the WNK serine-threonine kinase subfamily. Greater WNK1 expression associates with higher BP. A combination of promoters, enhancers, repressors, and insulators regulate WNK1 expression, but whether microRNAs also modulate WNK1 expression is unknown. Here, computational analysis revealed the presence of a target sequence for miR-192 and miR-215 at the same site in the 3' untranslated region of the ubiquitous L- and the kidney-specific KS-WNK1. We functionally validated this target sequence by transient transfection and reporter assays. Although we observed expression of both miRs along the distal nephron, only miR-192 regulated endogenous WNK1 ex vivo. Furthermore, a potassium load, sodium depletion, and aldosterone infusion each significantly reduced miR-192 expression in the kidney. Taken together, these results suggest a miR-driven mechanism of gene regulation by aldosterone and a role for miR-192 in the regulation of sodium and potassium balance in the kidney.

[Correlation study on serum adiponectin abnormity with adiponectin gene polymorphisms in hypertensive patients of phlegm-dampness constitution].

OBJECTIVE: To explore the difference of serum adiponectin (APN) level in hypertensive patients of phlegm-dampness constitution (PDC) and in those of non-PDC, as well as its association with APN gene polymorphisms. METHODS: Serum APN levels in 250 hypertensive patients (137 of PDC and 113 of non-PDC) were determined, and a correlation study was performed on 8 selected single nucleotide polymorphism (SNP) of APN gene. RESULTS: Significant differences of serum APN levels were observed between PDC and non-PDC patients (5.07 +/- 0.35 microg/mL vs 6.41 +/- 0.39 microg/mL, P = 0.045). No significant difference in polymorphism distribution of the 8 SNP sites of APN genes was found between patients of different constitutions (P > 0.05). Serum APN level was significantly lower in PDC patients than in non-PDC patients in sites of APN gene rs1063537 (3224C/T) polymorphism TT genotype (2.580 +/- 1.029 microg/mL vs 6.011 +/- 0.945 microg/mL, P = 0.017) and CT genotype (5.113 +/- 0.968 microg/mL vs 7.812 +/- 0.161 microg/mL, P = 0.021), while that of CC genotype was insignificant between the two constitutions (5.426 +/- 0.591 microg/mL vs 6.130 +/- 0.668 microg/mL). CONCLUSION: Serum APN level was significantly lower in hypertensive patients of PDC than in those of non-PDC. Moreover, the APN gene SNP3224 T allele carrier might be a hereditary feature of APN abnormity.

Deletion of WNK1 first intron results in misregulation of both isoforms in renal and extrarenal tissues.

Large deletions in intron 1 of the with-no-lysine kinase type 1 (WNK1) gene cause familial hyperkalemic hypertension. Alternative promoters generate functionally different isoforms: long ubiquitous isoforms (L-WNK1) and a kidney-specific isoform (KS-WNK1) lacking kinase activity. It remains unclear whether the disease-causing mutations selectively modify the synthesis of 1 or both types of isoforms. Using a transgenic mouse model, we found that intron 1 deletion resulted in the overexpression of L- and KS-WNK1 in the distal convoluted tubule and ubiquitous ectopic KS-WNK1 expression. Phylogenetic and functional analysis of the minimal 22-kb intron 1 deletion identified 1 repressor and 1 insulator, potentially preventing interactions between the regulatory elements of L-WNK1 and KS-WNK1. These results provide the first insight into the molecular mechanisms of WNK1-induced familial hyperkalemic hypertension.

Study on insulin resistance and genetic polymorphisms in essential hypertension patients of two different kinds of TCM constitution.

OBJECTIVE: To investigate the relationship of insulin resistance and the polymorphisms of insulin receptor-related genes in essential hypertension patients of two different kinds of TCM constitution. METHODS: Oral glucose tolerance test (OGTT) and insulin release test (InRT) were conducted in 217 essential hypertensive patients of either sluggish meticulous (SM) constitution (139 cases) or prosperous impetuous (PI) constitution (78 cases), and the polymorphism of three genes, including insulin-like growth factor-1 receptor (IGF-1R), insulin receptor substrate-1 (IRS-1) and 2 (IRS-2) genes were detected. RESULTS: (1) OGTT, InRT and insulin resistance index (Homa-IR) were higher and insulin sensitive index (ISI) was lower in the patients of SM constitution than those in patients of PI constitution. (2) Significant difference of ISI and Homa-IR was shown in patients of both constitutions with genotype G of the 3 genes. CONCLUSION: Decrease of insulin sensitivity and increase of insulin resistance are more obvious in hypertensive patients with genotype G of the 3 genes of SM constitution than in those of PI constitution. Therefore, the difference in constitution might be one of the genetic characteristics for insulin resistance in hypertensive patients.

[Effect of dihydroartemisinin on the ultrastructure of Trichomonas vaginalis trophozoites in vitro].

OBJECTIVE: To observe the effect of dihydroartemisinin (DHA) on the ultrastructure of Trichomonas vaginalis cultured in vitro. METHODS: The trophozoites of T. vaginalis were cultivated with liver extract solution medium containing 1 mg/ml dihydroartemisinin, and were then observed by scanning and transmission electron microscopes after the treatment for 2.5-4.0 h. RESULTS: The cell membranes of the trophozoites treated with DHA were damaged considerably. The surface of T. vaginalis showed deepening folds, hollow, and cracks. The nuclear membrane appeared fractures. There were a few of crevices in the nucleus and cytoplasm. Disordered and dilated endoplasmic reticulum, injured and deformed hydrogenosomes were found. Cytoplasm of the damaged parasites spilled over from torn place. After the cell membrane was peeled off, the nuclei, hydrogenosomes and pelta were exposed, which finally resulted in the death of the denatured parasites. CONCLUSION: Dihydroartemisinin can destroy membrane structure and organelles of Trichomonas vaginalis trophozoites, which leads to decomposition and necrosis of the parasites.