The rhizomes of Atractylodes macrocephala Koidz improve gastrointestinal health and pregnancy outcomes in pregnant mice via modulating intestinal barrier and water-fluid metabolism.

ETHNOPHARMACOLOGICAL RELEVANCE: Baizhu (BZ) is the dried rhizome of Atractylodes macrocephala Koidz (Compositae), which invigorates the spleen, improves vital energy, stabilizes the fetus, and is widely used for treating spleen deficiency syndrome. However, the impact of BZ on gastrointestinal function during pregnancy remains unexplored. AIM OF THE STUDY: This study elucidated the ameliorative effects of BZ on gastrointestinal health and pregnancy outcomes in pregnant mice with spleen deficiency diarrhea (SDD). METHODS: To simulate an irregular human diet and overconsumption of cold and bitter foods leading to SDD, a model of pregnant mice with SDD was established using an alternate-day fasting and high-fat diet combined with oral administration of Sennae Folium. During the experiment, general indicators and diarrhea-related parameters were measured. Gastric and intestinal motility (small intestinal propulsion and gastric emptying rates) were evaluated. Serum motilin (MTL), ghrelin, growth hormone (GH), gastrin (Gas), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-c), chorionic gonadotropin ß (ß-CG), progesterone (P), and estradiol (E2) were quantified using an enzyme-linked immunosorbent assay. Pathological changes were examined by hematoxylin and eosin staining (H&E) and alcian blue periodic acid Schiff staining (AB-PAS). Immunohistochemistry and immunofluorescence were used to measure the expression levels of the intestinal barrier and water metabolism-related proteins in colonic tissues. The pregnancy rate, ovarian organ coefficient, uterus with fetus organ coefficient, small size, average fetal weight, and body length of fetal mice were calculated. RESULTS: The results showed that BZ significantly improved general indicators and diarrhea in pregnant mice with SDD, increased gastric emptying rate and small intestinal propulsion rate, elevated the levels of gastrointestinal hormones (AMS, ghrelin, GH, and Gas) in the serum, and reduced lipid levels (TC and LDL-c). It also improved colonic tissue morphology, increased the number of goblet cells, and promoted the mRNA and protein expression of occludin, claudin-1, ZO-1, AQP3, AQP4, and AQP8 in colonic tissues, downregulating the mRNA and protein expression levels of claudin-2, thereby alleviating intestinal barrier damage and regulating the balance of water and fluid metabolism. BZ also held the levels of pregnancy hormones (ß-CG, P, and E2) in the serum of pregnant mice with SDD. Moreover, it increased the pregnancy rate, ovarian organ coefficient, uterus with fetus organ coefficient, litter size, average fetal weight, and body length of fetal mice. These findings indicate that BZ can improve spleen deficiency-related symptoms in pregnant mice before and during pregnancy, regulate pregnancy-related hormones, and improve pregnancy outcomes.

Preparative Purification of Linarin Extracts from Dendranthema indicum Flowers and Evaluation of Its Antihypertensive Effect.

Background. Preliminary research showed that linarin (LIN) might have a relationship with the antihypertensive effect of Dendranthema indicum flowers. However, the preparative method for LIN enriched extract from Dendranthema indicum flowers was not clear and its antihypertensive effect was not confirmed. In this study, we designed a series of experiments to develop an efficient method for purification of LIN extracts and confirm the possibility of LIN extracts to be an antihypertensive drug. Materials and Methods. HPLC-VWD/DAD were used in the process of developing purification method. The antihypertensive effect of LIN extracts was tested by CODA Mouse & Rat Tail-Cuff Blood Pressure System; western blot and biochemical analysis were used to investigate mechanism and toxicity. Results. The content and recovery of LIN reached 55.68 ± 2.08% and 66.65 ± 1.73%, respectively, through solid-liquid extraction. The composition of product was stable through the analysis of fingerprint. Chronic administration of LIN extracts reduced blood pressure obviously which had a relationship with the inhibition of renin-angiotensin system (RAS) in kidney and the function indexes of kidney and liver had no variations. Conclusions. The preparation method was simple, low-cost, and stable, and it was fit for industrial application. The LIN prepared by this method had the potential to be an antihypertensive drug.

A new fluorescent quantitative polymerase chain reaction technique.

To perform real-time detection of specific genes, a new complex probe has been designed and synthesized. Based on fluorescence resonance energy transfer (FRET), this complex probe is composed of a long-fluorescent reporter probe and a short-quenching probe. The 5' end of the fluorescent probe is connected to a fluorescein molecule, and its 3' end is linked to an extending blocking molecule. The 3' end of the quenching probe is connected to a quenching molecule-p-methyl red (Dabcyl). The quenching probe is complementary to the 5' end of the fluorescent probe. When there is no template, the two probes combine to form a complex probe and therefore no fluorescence is produced; when there are templates, the fluorescent probe hybridizes with the templates first, and the fluorescence is not quenched. The fluorescence intensity produced is in direct proportion to the template quantity. In accordance with the principles of reaction of the complex probe, we have studied the probe's FRET nature and the factors that affect it, including the quenching probe and amplified fragment length, the proper proportion of the fluorescent probe to the quenching probe, and the magnesium ion concentration. Experimental results showed that the quenching probe and its amplified fragment length had an obvious impact on the function of the complex probe. The quenching probe used in the present experiment is up to 21 nucleotides long, with an amplified fragment of 127bp. The most preferable reaction system is obtained when the proportion of the fluorescent probe to the quenching probe is 1:1, and the concentration of magnesium ions is 3mmol/L. The complex probe is easy to synthesize. The quenching is thorough with good accuracy and specificity. The sensitivity reaches 10(2) copies with a very large dynamic quantitation range. Accurate quantitation can be achieved with samples detected within 10(2)-10(9) copies. The complex probe method can be used to detect virus infection levels, transgenic copy quantities, single nucleotide polymorphisms, etc.