[Assay of amino acids in leaves of Eucommia ulmoides under arbor forest mode and leaf-oriented cultivation mode by pre-column derivatization HPLC].

This study aims to develop the pre-column derivatization high performance liquid chromatography(HPLC) method for the determination of 16 kinds of amino acids in Eucommia ulmoides leaves, and compare the content of amino acids in the leaves harvested at different time and under leaf-oriented cultivation mode(LCM) and arbor forest mode(AFM). The HPLC conditions are as below: phenyl isothiocyanate(PITC) as pre-column derivatization agent, Agilent ZORBAX C_(18 )column(4.6 mm×250 mm, 5 µm), mobile phase A of acetonitrile-water(80∶20), mobile phase B of 0.1 mol·L~(-1) sodium acetate solution-acetonitrile(94∶6), gradient elution, flow rate of 1.0 mL·min~(-1), injection volume of 5 µL, column temperature of 40 ℃, and detection wavelength of 254 nm. The HPLC profile indicated well separation of 16 kinds of amino acids and the amino acid content in E. ulmoides leaves was up to 16.26%. In addition, the amino acid content in leaves of E. ulmoides under LCM was higher than under AFM. The amino acid content varied with the harvesting time. Through orthogonal partial least squares discriminant analysis, the amino acids of E. ulmoides under LCM and AFM were compared, which can distinguish the leaves under LCM from those under AFM. Principal component analysis was applied to comprehensively score the amino acids of E. ulmoides leaves. The results showed that the score of leaves under LCM was higher than that under AFM. Nutritional evaluation results indicated that the proteins in E. ulmoides leaves belonged to high-quality vegetable proteins. The established method for the determination of amino acid content is reliable. With the amino acid content as index, the leaf quality of E. ulmoides under LCM is better than that under AFM. This study lays a theoretical basis for the promotion of LCM for E. ulmoides and the development of medicinal and edible products from E. ulmoides leaves.

Erianin Exerts Antineoplastic Effects on Esophageal Squamous Cell Carcinoma Cells by Activating the cGMP-PKG Signaling Pathway.

Erianin, a natural compound extracted from Dendrobium chrysotoxum Lindl, has potential therapeutic benefits against various tumors. However, its role in esophageal squamous cell carcinoma (ESCC) remains unclear. Cell proliferation was analyzed by CCK8, colony-formation, and EdU proliferation assays, while cell migration was evaluated through wound healing assays as well as determination of the protein expression levels of epithelial-to-mesenchymal transition (EMT) markers and ß-catenin. Apoptosis was measured by flow cytometry. RNA sequencing (RNA-seq) and bioinformatic analyses were performed to elucidate the underlying mechanisms of erianin in ESCC. Enzyme-linked immunosorbent assay (ELISA) was used to determine intracellular cGMP, cleaved-PARP, and caspase-3/7 activity, while mRNA and protein levels were determined by qRT-PCR and western blotting, respectively. Our results indicate that erianin significantly inhibited ESCC cell proliferation and migration while also promoting apoptosis. Mechanistically, RNA sequencing coupled with KEGG enrichment analysis and functional assays revealed that activation of the cGMP-PKG pathway contributed to the antitumor effects of erianin, whereas the c-GMP-dependent protein kinase inhibitor KT5823 significantly attenuated these effects. In conclusion, our results demonstrate that erianin suppresses the proliferation of ESCC cells by activating the cGMP-PKG pathway, suggesting that erianin could be a promising candidate for the treatment of ESCC.

Study on the correlation between vaginal bleeding in first trimester and preterm birth: A birth cohort study in Lanzhou, China.

OBJECTS: To investigate the correlation between first trimester vaginal bleeding and preterm birth (PB), and to offer suggestions on the perinatal health care and preterm birth prevention. METHODS: A birth cohort study was conducted on 10 179 pregnant women. Unconditional logistic regression model was used to evaluate the associations between vaginal bleeding and preterm birth in sub-preterm groups. RESULTS: Of the 10 179 pregnant women included, a total of 1001 women suffered from vaginal bleeding during the first trimester, of which 119 suffered from PB. Any vaginal bleeding increased the risk of PB. Severe bleeding was a high-risk factor of PB, associated with 4.8-fold risk of very PB, 2.7-fold risk of spontaneous PB without PROM (premature rupture of membrane) and 4.6-fold risk of medical induced PB. Bleeding prolonged more than 1 week increased 66% risk of PB and 36% risk of PB on initial episode happened in 5-12 weeks of gestation age, especially in moderate PB, in medical-induced PB and in spontaneous PB with PPROM (preterm premature rupture of membrane which is one cause of PB). Mild bleeding or bleeding within 1 week or initial episode happened within 4 weeks of gestation age possibly had no influence on PB. CONCLUSION: Vaginal bleeding in the first-trimester was an independent risk factor for PB. The severity, duration and initial time of vaginal bleeding had different effects on different subtypes of PB.

Duck IL-2 promoter cloning and the effects of methylation status on mRNA levels in immune tissues.

Interleukin 2 (IL-2), a cytokine, plays an important role in animal immune systems. To investigate the influences of epigenetic modifications on transcription of the duck IL-2 gene, the promoter region of the duck IL-2 gene was cloned. Then, the DNA methylation status of the IL-2 gene promoter (-1337 bp/-924 bp) in immune tissues of ducks was determined using the Sequenom Mass Array methylation technique, and their corresponding expression levels were determined using real-time PCR. The results showed that 2850 bp of the duck IL-2 gene promoter region were obtained. There was one CpG island (-1231 bp/-902 bp) in which 11 CpG sites were distributed. The CpG1 and CpG2 sites are located between the binding sites of NFAT and AP-1, and they had higher homology methylation patterns in different individuals and tissues. The methylation frequencies of 28.5% CpG sites showed negative correlations with the expression levels of the IL-2 mRNA, whereas 71.5% showed positive correlations. These results indicate that the transcription of duck IL-2 may be distinct from that of mammals. CpG1 (-1284 bp) and CpG2 (-1264 bp) in the duck IL-2 promoter showed a higher homology of methylation patterns, indicating a similar regulatory effect on their gene expression, and these CpG sites may be essential for the regulation of transcription of duck IL-2. The methylation pattern of the IL-2 gene promoter in duck was tissue specific.

The research for the clinical curative effect through combing traditional Chinese medicine with insulin to cure diabetes.

UNLABELLED: The clinical curative effect is observed through curing type 2 diabetes mellitus with the therapy of combing Traditional Chinese Medicine (TCM) with insulin. METHOD: Both the insulin prescription and the treatment of traditional Chinese medicine prescription are applied as mutual comparison. And the dosage, time, blood sugar level and curative effect etc are recorded. Healthy human body is taken as comparison for monitoring physical indicators. RESULTS: Through comparing insulin prescription and the combing therapy of insulin and traditional Chinese medicine, the insulin treatment group is better than contrast group (P<0.05). For the blending use group, the ISI in each group is significantly lower than that of health control group (P<0.01), where accumulation of damp heat in spleen type is the lowest; the BM I, H bA1C of type 2 diabetic patient is higher than health control group, its accumulation of damp heat in spleen type is the highest, TC, TG typical accumulation of damp heat in spleen are higher than other pattern of syndrome. CONCLUSION: the treatment method of combing TCM with insulin in curing type 2 diabetes mellitus has better effect than using insulin treatment alone; the resistance degree of insulin demonstrates the changing trend of first increase and later decrease with the development of disease course. Accumulation of damp heat in spleen type accounts for the highest proportion in type 2 diabetic patients, and there exists serious insulin resistance.