Metabolomics-based comparative study of breast colostrum and mature breast milk.

Breast milk is the safest and most complete natural food for babies. Although breast milk is crucial to the health and development of infants, the metabolites in breast milk during lactation period have not been characterized. Therefore, we examined and compared the metabolites in breast colostrum and mature breast milk using gas chromatography-time-of-flight-mass spectrometry-based metabolomics. A total of 159 metabolites were characterized, of which 72 were differentially expressed metabolites (DEMs), including 17 upregulated and 55 downregulated DEMs in breast colostrum compared to those in mature breast milk. Metabolic pathway analysis revealed that these DEMs were related to glycine, serine, and threonine metabolism; glyoxylate and dicarboxylate metabolism; alanine, aspartate, and glutamate metabolism; pentose and glucuronate interconversions; and aminoacyl-tRNA biosynthesis. Our results improve the understanding of breast milk composition and provide a theoretical basis for optimizing infant formula to closely imitate the nutrients required for proper growth and development of babies.

Huayu Tongluo Recipe Attenuates Renal Oxidative Stress and Inflammation through the Activation of AMPK/Nrf2 Signaling Pathway in Streptozotocin- (STZ-) Induced Diabetic Rats.

Diabetic nephropathy (DN), a severe microvascular complication of diabetes, is one of the leading causes of end-stage renal disease. Huayu Tongluo Recipe (HTR) has been widely used in the clinical treatment of DN in China, and its efficacy is reliable. This study aimed to explore the renoprotective effect of HTR and the underlying mechanism. Male Sprague-Dawley rats were fed with high sugar and fat diet combined with an intraperitoneal injection of STZ to establish the diabetic model. Rats in each group were respectively given drinking water, HTR, and irbesartan by gavage for 16 weeks. 24-hour urine samples were collected every 4 weeks to detect the content of total protein and 8-OHdG. Blood samples were taken to detect biochemical indicators and inflammatory markers at the end of 16th week. Renal tissue was collected to investigate pathological changes and to detect oxidative stress and inflammatory markers. AMPK/Nrf2 signaling pathway and fibrosis-related proteins were detected by immunohistochemistry, immunofluorescence, real-time PCR, and western blot. 24h urine total protein (24h UTP), serum creatinine (Scr), blood urea nitrogen (BUN), total cholesterol (TC), and triglyceride (TG) were decreased in the rats treated with HTR, while there was no noticeable change of blood glucose. HTR administration decreased malondialdehyde (MDA) content and increased superoxide dismutase (SOD) activity in kidneys, complying with reduced 8-OHdG in the urine. The levels of TNF-α, IL-1ß, and MCP1 and the expression of nuclear NFκB were also lower after HTR treatment. Furthermore, HTR alleviated pathological renal injury and reduced the accumulation of extracellular matrix (ECM). Besides, HTR enhanced the AMPK/Nrf2 signaling and increased the expression of HO-1 while it inhibited the Nox4/TGF-ß1 signaling in the kidneys of STZ-induced diabetic rats. HTR can inhibit renal oxidative stress and inflammation to reduce ECM accumulation and protect the kidney through activating the AMPK/Nrf2 signaling pathway in DN.

New cytotoxic biflavones from Selaginella doederleinii.

Three new biflavones, apigenin-(3',8″)-chrysin (1), (2S)-2,3-Dihydroametoflavone 5,4'-dimethyl ether (2), and (2S)-5″,7″-Dihydroxy-2″-phenoxychromonyl-(4'″,3')-naringenin (3), together with seven known biflavones (4-10) were isolated from the 75% EtOH extract of Selaginella doederleinii. The structures of new compounds were established by application of spectroscopic methods, including 1D and 2D NMR, HRMS, and CD measurements. In addition, all new compounds were evaluated for their cytotoxic potential against three human cancer cell lines A549, MCF-7, and SMMC-7721 in vitro. Compound 2 exhibited potent cytotoxic activity with IC50 values ranging from 6.35 to 10.18 µM.

New neolignans from Selaginella picta and their protective effect on HT-22 cells.

Three new compounds, pictalignans A (1), B (2), C (3), along with three known analogues, syringaresinol (4), 3,3',5-trimethoxy-4',7-epoxy-8,5'-neoligan-4',9,9'-triol (5), 4,9-dihydroxy-4',7-epoxy-8',9'-dinor-8,5'-neolignan-7'-oic acid (6) were isolated from the 75% aqueous ethanol extract of Selaginella picta. Their structures were established by physicochemical properties and spectroscopic methods, and absolute configurations of new compounds were elucidated by experimental and calculated ECD spectra. Compounds 1-3 are neolignans with additional one or two C6-C3 structural units attached to hydroxypropyl group, which are extremely rare in nature. All new compounds exhibited moderate protective effect against the injury of HT-22 cells induced by L-Glutamate in vitro, and compound 1 showed better protective effect than positive drug with the concentrations of 10 µM to 15 µM.

Cloning and characterizing of the murine IRF-3 gene promoter region.

The interferon regulatory factor 3 (IRF-3) plays essential roles in inflammation and immune response. Here, we cloned the nucleotide sequence of the 5'-flanking region of the murine IRF-3 gene (mIRF-3) and characterized the molecular mechanisms controlling the mIRF-3 transcriptional activity in NIH3T3 cells. Analyses of a series of 5' deletion constructs demonstrated that a 301 bp region (-255/+46) of the mIRF-3 gene is sufficient for full promoter activity. This region contains IK1, Egr2, Cmyb, E2F1 and YY1 putative transcription factor binding sites. Mutation of Egr2 or YY1 site led to 52-68 % decrease of the mIRF-3 promoter activity, and double Egr2 and YY1 mutation reduced the promoter activity to 20 % of the wild-type promoter activity. Furthermore, knockingdown of endogenous Egr2 or YY1 by a siRNA strategy markedly inhibited the mIRF-3 promoter activity. Chromatin immunoprecipitation assays showed that Egr2 and YY1 interact with the mIRF-3 promoter in vivo. These results suggested that the basal promoter activity of the mIRF-3 gene is regulated by transcription factors Egr2 and YY1 in NIH3T3 cells.

[Regulatory Effect of Chinese Drugs for Stasis Removing and Collaterals Dredging on the Expres- sions of podocin and CD2AP in Podocyte Slit Diaphragm of Diabetic Nephropathy Rats].

OBJECTIVE: To observe the regulatory effect of Chinese drugs for stasis removing and collaterals dredging (CDSRCD) on the expressions of podocin and CD2AP in podocyte slit diaphragm (SD) of diabetic nephropathy (DN) rats. METHODS: DN rat model was duplicated in 40 male Sprague- Dawley rats by feeding high fat high glucose diet combined with intraperitoneally injecting 1 % streptozoto- cin (STZ, 35 mg/kg). Totally 36 successfully modeled rats were divided into the model group, the CD- SRCD group,- and the irbesartan group according to random digit table, 12 in each group. Besides, anoth- er 10 normal rats were recruited as a normal group. Rats in the CDSRCD group and the irbesartan group were intragastrically fed with CDSRCD and irbesartan respectively. Rats in the normal group and the mod- el group were fed with equal volume of distilled water at the same time. 24 h urine protein quantitation was detected using ELISA at various time points. Body weight (BW) , kidney weight ( KW), kidney index (KI) , fasting blood glucose (FBG) , serum creatinine (SCr), blood urea nitrogen (BUN), and uric acid (UA) in each group were detected after 16 weeks of intervention. The pathomorphological changes of re- nal tissue were observed under light microscope and electron microscope respectively. The protein and mRNA expressions of podocin and CD2AP were detected by Western blot and Real-time PCR respectively. RESULTS: (1) Compared with the normal group, 24 h urine protein quantitation significantly increased at week 4, 8, 12, and 16, respectively (P <0. 01). BW was decreased; KI and levels of FBG, SCr, BUN, and UA all increased after modeling (P <0. 01). Compared with the model group, 24 h urine protein quan- titation significantly decreased in the CDSRCD group and the irbesartan group at week 4, 8, 12, and 16, respectively (P <0. 01). Besides, it was more obviously reduced in the CDSRCD group than in the irbe- sartan group (P <0. 05, P <0.01). BUN level obviously decreased both in the CDSRCD group and the irbesartan group after modeling (P <0. 05, P <0. 01). (2) Results of renal pathology showed that disar- ranged renal structure, obviously thickened basement membrane, severely proliferated mesenteria, widely fused foot processes in the model group. All these pathological changes were attenuated in the CD- SRCD group and the irbesartan group to some degree. (3) Results of Western blot and Real-time PCR showed, compared with the normal group, protein and mRNA expressions of podocin and CD2AP decreased in the model group (P <0. 01). Compared with the model group, protein and mRNA expressions of podocin and CD2AP increased in the CDSRCD group and the irbesartan group (P <0. 01). Protein and mRNA expressions of podocin and CD2AP increased more in the CDSRCD group than in the irbesartan group (P <0. 05). CONCLUSIONS: CDSRCD could protect renal function by lowering urinary protein in DN rats, improve renal pathological changes. Its mechanism might be related to up-regulating mRNA and protein expressions of podocin and CD2AP.

Expression of ascorbate peroxidase and glutathione reductase in roots of rice seedlings in response to NaCl and H2O2.

The accumulation of H2O2 by NaCl was observed in the roots of rice seedlings. Treatment with NaCl caused an increase in the activities of ascorbate peroxidase (APX) and glutathione reductase (GR) and the expression of OsAPX and OsGR in rice roots. Exogenously applied H2O2 also enhanced the activities of APX and GR and the expression of OsAPX and OsGR in rice roots. The accumulation of H2O2 in rice roots in response to NaCl was inhibited by the NADPH oxidase inhibitors, diphenyleneiodonium chloride (DPI) and imidazole (IMD). However, DPI, IMD, and dimethylthiourea, a H2O2 trap, did not reduce NaCl-enhanced activities of APX and GR and expression of OsAPX and OsGR. It appears that H2O2 is not involved in the regulation of NaCl-induced APX and GR activities and OsAPX and OsGR expression in rice roots.