High and low dose of luzindole or 4-phenyl-2-propionamidotetralin (4-P-PDOT) reverse bovine granulosa cell response to melatonin.

Background: Communication between oocytes and granulosa cells ultimately dictate follicle development or atresia. Melatonin is also involved in follicle development. This study aimed to investigate the effects of melatonin and its receptor antagonists on hormone secretion, as well as gene expression related to hormone synthesis, TGF-ß superfamily, and follicle development in bovine granulosa cells, and assess the effects of melatonin in the presence of 4-P-PDOT and luzindole. Methods: Bovine ovaries were collected from a local abattoir and follicular fluid (follicle diameter 5-8 mm) was collected for granulosa cell isolation and culture. Granulosa cells and culture medium were collected 48 h after treatment with melatonin at high dose concentrations (10-5 M) and low dose concentrations (10-9 M) in the absence/presence of 4-P-PDOT and luzindole (10-5 M or 10-9 M). Furthermore, the expression level of genes related to hormonal synthesis (CYP11A1, CYP19A1, StAR, and RUNX2), TGF-ß superfamily (BMP6, INHA, INHBA, INHBB, and TGFBR3), and development (EGFR, DNMT1A, and FSHR) were detected in each experimental group by real-time quantitative PCR. In addition, the level of hormones in culture medium were detected using ELISA. Results: Both 10-5 M and 10-9 M melatonin doses promoted the secretion of inhibin A and progesterone without affecting the production of inhibin B and estradiol. In addition, both promoted the gene expression of INHA, StAR, RUNX2, TGFBR3, EGFR, and DNMT1A, and inhibited the expression of BMP6, INHBB, CYP11A1, CYP19A1, and FSHR. When combined with different doses of 4-P-PDOT and luzindole, they exhibited different effects on the secretion of inhibin B, estradiol, inhibin A, and progesterone, and the expression of CYP19A1, RUNX2, BMP6, INHBB, EGFR, and DNMT1A induced by melatonin. Conclusion: High and low dose melatonin receptor antagonists exhibited different effects in regulating hormone secretion and the expression of various genes in response to melatonin. Therefore, concentration effects must be considered when using luzindole or 4-P-PDOT.

Genes and Signaling Pathways Involved in the Regulation of Selenium-Enriched Yeast on Liver Metabolism and Health of Broiler (Gallus gallus).

Selenium-enriched yeast (SeY) plays an important role in the liver health and metabolism of the broiler. However, the mechanism by which it regulates liver metabolism and the health of broilers is largely unknown. Therefore, this study was conducted to elucidate the key genes and signaling pathways involved in regulating SeY in liver metabolism and bird's health. Thus, the mRNA expression microarray, GSE25151, was downloaded from Gene Expression Omnibus (GEO) database. GSE25151 consists of liver samples from SeY-treated and the control broilers. Six hundred four differentially expressed genes (DEGs) were identified in livers between SeY-treated and control. Gene ontology (GO) enrichment analysis indicated that those DEGs are mainly involved in metabolism-related biological processes, such as biological regulation, molecular processes, responses to stimuli, cell communication and proliferation, and growth. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed the DEGs mainly enriched in metabolism-related signaling pathways, including PI3K, Akt, Wnt, calcium, IGF1 receptor, and MAPK signaling pathways. Moreover, many genes, such as NMUR1, NMU, and GPRC6A, might contribute to the regulation of SeY to broiler liver metabolism and health. In conclusion, the current study enhances our understanding of the regulation of SeY in liver metabolism and health of the birds and will assist studies of the molecular mechanisms of SeY regulation in chicken liver.

New insights on the analysis of phytosterols in pollen and anther wall of tree peony (Paeonia ostii 'Fengdan') revealed by GC-MS/MS.

Phytosterols exhibit a wide range of pharmacological activities and have no side-effects, which have attracted more and more attention. In this study, two pre-treatment methods (alkaline hydrolysis and acid-alkali hydrolysis) were performed in parallel for the first time to investigate the effects on phytosterol analysis. The aim is to develop an analytical method for phytosterols in pollen and anther wall of tree peony (Paeonia ostii 'Fengdan') by gas chromatography-mass spectrometry. The result indicated that the contents of phytosterols in pollen (3390.02 mg/100 g DW) and anther wall (929.70 mg/100 g DW) of P. ostii 'Fengdan' were relatively high. Meanwhile, eleven phytosterols were identified, among which five phytosterols were first identified in tree peony pollen. Moreover, the effects of wall breaking and different developmental stages on phytosterol contents of pollen and anther wall of P. ostii 'Fengdan' were also discussed. The result showed that wall breaking was beneficial to sterol detection of pollen, but not necessary for anther wall. For the best harvest time, the stage of S2 had the highest content of phytosterols. In conclusion, this study successfully establishes an analytical method for phytosterols by gas chromatography-mass spectrometry, and lays a foundation for the development and utilization of pollen and anther wall of P. ostii 'Fengdan' in functional foods and pharmaceutical industry.

Comprehensive Effect of Oxidant Addition in an FGD Slurry on the Removal and Distribution of Selenium: A Field Study.

Flue gas desulfurization (FGD) scrubbers capture selenium in coal-fired power plants, leading to a high concentration of selenium in the slurry. This research proves that SO32- is preferentially oxidized compared to SeO32- by S2O82-. With the increase in the oxidation-reduction potential (ORP) caused by S2O82- addition, the conversion rate of SO32- increased and the size of gypsum grains grew from 31.2 to 34.6 µm. SeO32- migrates into gypsum grains during the growth of CaSO4·2H2O, leading to selenium fixation in gypsum. In a field study of a 350 MW unit, the ORP increased from 142 to 450 mV when Na2S2O8 was fed into the FGD slurry. With the addition of the oxidant, 65.1% of selenium in the liquid phase migrated into gypsum. The concentration of selenium in the leachate of gypsum after oxidant addition decreased by 68.0%. A 2.34% increase in the selenium removal rate was observed in the scrubber. This study focuses on the migration and conversion of selenium in an actual FGD slurry via a field test. The results found in the 350 MW unit are consistent with laboratory results. The change in ORP has been proven to be effective in adjusting the selenium distribution in the FGD slurry.

De novo Splice Site Mutation of the CHD7 Gene in a Chinese Patient with Typical CHARGE Syndrome.

INTRODUCTION: CHARGE syndrome (CS, OMIM 214800) is a rare genetic disease characterized by multiple congenital abnormalities, including coloboma, heart defect, atresia of the choanae, retardation of development, genital anomalies, and ear anomalies/deafness. The syndrome is mainly caused by a heterozygous variant in the chromodomain helicase DNA-binding protein 7 (CHD7) gene that encodes the CHD7 protein, involved in the ATP-dependent remodeling of chromatin. METHODS: In this study, the next-generation sequencing targeted panel was used to detect a de novo variant c.3523-2A>G in the CHD7 gene in a patient with severe CS, congenital heart disease, left coloboma of the choroid, cryptorchidism, and congenital deafness. The Sanger sequencing confirmed the variant and clarified it as de novo variant by short tandem repeat analysis in the patient family. We analyzed the effect of a variant by Minigene assay to evaluate the pathogenicity of the variant. RESULTS: In summary, cDNA analysis confirmed that c.3523-2A>G variant activates a cryptic splice site, resulting in 172 base pair missing in exon 15, leading to the premature truncation of the CHD7 protein (p.V1175Afs*11). CONCLUSION: The present study functionally characterized the novel c.3523-2A>G variant in CHD7, providing further confirmatory evidence that it is associated with CS.

Selenium migration mechanism in wet FGD slurry: Experimental and DFT analysis.

Selenium (Se) is one of the hazardous trace elements emitted from coal-fired power plants. The Se migration behavior in wet flue gas desulfurization (FGD) slurry is still unclear, and the species of Se in FGD gypsum remains controversial. In this research, the bubbling experiments using simulated slurry with/without gypsum crystallization process were conducted. The experimental results indicated that pure gypsum has poor capability to capture Se components, and only selenite could be trapped in gypsum during its crystal growth stage. Furthermore, the DFT calculation was conducted to provide the microscopic information of Se adsorption and substitution characteristics during gypsum crystallization process. The research findings of this study could help understand the mechanism of Se migration process in FGD slurry, and facilitate the development of effective Se emission control technologies in the future.

Effect of Combining Early Chemotherapy with Zhipu Liujunzi Decoction under the Concept of Strengthening and Consolidating Body Resistance for Gastric Cancer Patients and Nursing Strategy.

Objective: To explore the clinical efficacy of combining early chemotherapy with Zhipu Liujunzi decoction under the concept of strengthening and consolidating body resistance for gastric cancer patients and nursing strategy. Methods: The clinical data of 100 patients undergoing radical gastrectomy in our hospital from July 2019 to July 2020 were selected for the retrospective analysis, and the patients were divided into the control group and experimental group according to different treatment methods, with 50 cases in each group. Early chemotherapy after surgery was given to patients in the control group, and on the basis of the aforesaid treatment and under the concept of strengthening and consolidating body resistance, patients in the experimental group took Zhipu Liujunzi decoction and received the nursing strategy, so as to compare their effective rate, adverse reaction rate (ARR), immune function indicators, KPS scores, and nursing satisfaction scores. Results: After treatment, the experimental group obtained significantly higher objective remission rate (ORR) and disease control rate (DCR) (P < 0.05), lower carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) levels (P < 0.001), higher immune parameters levels (P < 0.001), higher KPS scores and lower TCM symptom scores (P < 0.001), lower PSQI scores, SAS scores, and SDS scores (P < 0.001) and higher nursing satisfaction scores (P < 0.001), and lower total accidence rate of toxic side effects (P < 0.05) than the control group. Conclusion: Under the concept of strengthening and consolidating body resistance, combining early chemotherapy with Zhipu Liujunzi decoction is a reliable method for improving the immune function and quality of life for gastric cancer patients with higher safety. Such a strategy greatly reduces the tumor marker levels in patients. Further research will be conducive to establishing a better solution for gastric cancer patients.

Effect Observation on Modified Zishen Tongguan Decoction Combined with Acupuncture in Treatment of Urinary Retention after Cervical Cancer Surgery and Its Influence on the Incidence of Adverse Reactions.

OBJECTIVE: To explore the effect observation on modified Zishen Tongguan decoction combined with acupuncture in the treatment of urinary retention after cervical cancer surgery and its influence on the incidence of adverse reactions. METHODS: The clinical data of 84 patients suffered from urinary retention after radical resection of cervical cancer (December 2018-December 2019) in the oncology department of Jinan Municipal Hospital of Traditional Chinese Medicine were selected for retrospective analysis. According to the order of admission, they were divided into group A (n = 42), treated with conventional therapy, modified Zishen Tongguan decoction, and acupuncture, and group B (n = 42), treated with conventional therapy. The clinical efficacy of the two groups was observed, the urination function indexes after therapy were recorded, and the clinical efficacy and incidence of adverse reactions were analyzed. RESULTS: After therapy, compared with group B, the average urinary flow rate, maximum urinary flow rate, bladder compliance (BC) level value, and the number of patients with good recovery of bladder function of group A were obviously higher (P < 0.05), and the urination time and detrusor pressure were obviously lower (P < 0.001). There was no significant difference in the average scoring of overactive bladder syndrome score (OABSS) between the two groups at 7 days of therapy (p > 0.05). The average OABSS of group A at 14 days of therapy was obviously lower than that of group B (P < 0.001). Compared with group B, the total clinical effective rate of group A was obviously higher (P < 0.05), while the total incidence of adverse reactions was obviously lower (P < 0.05). CONCLUSION: Modified Zishen Tongguan decoction combined with acupuncture is a reliable method to treat urinary retention after cervical cancer surgery, which greatly improves the urination function of patients, as well as the clinical efficacy. Further research will help create a better solution for patients with urinary retention after cervical cancer surgery.

Luzindole and 4P-PDOT block the effect of melatonin on bovine granulosa cell apoptosis and cell cycle depending on its concentration.

Granulosa cells play an essential physiological role in mediating the follicle development and survival or apoptosis of granulosa cells dictate the follicle development or atresia. The aim of this study was to investigate the role of high dose (10-5 M) and low dose (10-9 M) melatonin in bovine granulosa cells, and assess whether MT1 and MT2 inhibiter affect granulosa cells response to melatonin. We found that the high dose (10-5 M) and low dose (10-9 M) both could act as an essential role in modulating granulosa cells apoptosis, cell cycle and antioxidant. The beneficial effect could be related to that melatonin promoted the expression of Bcl2, Bcl-xl, SOD1 and GPX4, and inhibited Bax, caspase-3 and p53 expression. Moreover P21 expression was decreased in granulosa cells treated with the high dose (10-5 M) melatonin and increased in that treated with the low dose (10-9 M) melatonin. To further reveal the role of MT1 and MT2 in mediating the effect of melatonin on granulosa cells apoptosis, cell cycle and antioxidant, we found that the luzindole and 4P-PDOT did not affect the effect of high dose (10-5 M) melatonin on regulating Bcl2, Bax, caspase-3, SOD1, GPX4 and p53 expression, while blocked its effect on modulating Bcl-xl and P21expression. However, luzindole and 4P-PDOT disturbed the effect of low dose (10-9 M) melatonin on regulating Bcl2, Bax, caspase-3, Bcl-xl, SOD1, GPX4, and p53 expression. In conclusion, these results reveal that the effect of low dose (10-9 M) melatonin on granulosa cells apoptosis are mediated by MT1 and MT2, and the high dose (10-5 M) melatonin affect the granulosa cells apoptosis by other pathway, besides MT1 and MT2. Moreover MT1 and MT2 may work in concert to modulate bovine granulosa cells function by regulating cellular progression and apoptosis.

Pterostilbene supplements carry the risk of drug interaction via inhibition of UDP-glucuronosyltransferases (UGT) 1A9 enzymes.

Pterostilbene (PT) is a natural stilbene common in small berries and food supplements, possessing numerous pharmacological activities. However, whether PT can affect the activities of UDP-glucuronosyltransferases (UGT) enzymes remains unclear. The aim of the present study was to investigate the effect of PT on UGT activities and to quantitatively evaluate the food-drug interaction potential due to UGT inhibition. Our data indicated that PT exhibited potent inhibition against HLM, UGT1A6, UGT1A9, UGT2B7, and UGT2B15, moderate inhibition against UGT1A1, UGT1A3, UGT1A8, and UGT2B4, negligible inhibition against UGT1A4, UGT1A7, UGT1A10, and UGT2B17. Further kinetic investigation demonstrated that PT exerted potent noncompetitive inhibition 4-MU glucuronidation by UGT1A9, with IC50 and Ki values of 0.92 µM and 0.52 ± 0.04 µM, respectively. Quantitative prediction study suggested that coadministration of PT supplements at 100 mg/day or higher doses may result in at least a 50% increase in the AUC of drugs predominantly cleared by UGT1A9. Thus, the coadministration of PT supplements and drugs primarily cleared by UGT1A9 may result in potential drug interaction, and precautions should be taken when coadministration of PT supplements and drugs metabolized by UGT1A9.

Posttreatment With LYRM03 Protects Rats From Acute Lung Inflammation Induced by Lipopolysaccharide via Suppressing the NF-κB/MyD88/TLR4 Axis.

BACKGROUND: 3-amino-2-hydroxy-4-phenyl-valyl-isoleucine (LYRM03) has been shown to be beneficial in a rat model of acute lung injury (ALI). Nonetheless, the pharmacologic action of LYRM03 interference has not been demonstrated to occur through oxidative stress and apoptosis in a rat lipopolysaccharide (LPS)-induced ALI model, and the potential pathogenic mechanism needs to be clarified. Our research intended to explore the mechanism of action using an in vivo rat LPS-induced ALI model and highlight the associated pathogenesis. METHODS: Sprague-Dawley rats were randomly assigned to the following five groups: Sham; LPS (5 mg/kg); LPS + LYRM03 (5 mg/kg); LPS + LYRM03 (10 mg/kg); and LPS + LYRM03 (20 mg/kg). Pulmonary injury indicators were documented at 24 h after LPS-induced ALI. Morphologic alterations, such as the extent of the injury, were determined using hematoxylin-eosin staining. Furthermore, expression levels of oxidative stress indicators (malondialdehyde, superoxide dismutase, and glutathione peroxidase) and inflammatory molecules (tumor necrosis factor-α, interleukin-8, and interleukin-6) in circulation were observed. The production of apoptosis-associated proteins (poly ADP-ribose polymerase, c-caspase 3, B-cell lymphoma-2, and Bcl2 associated X), inflammatory mediators (high mobility group box-1, toll-like receptor 4, nuclear factor-kappa B p65, and myeloid differentiation primary response 88), and inhibitor of kappa B-α were determined through Western blotting. Real-time polymerase chain reaction was applied to assess the messenger RNA expression of the inflammatory mediators. RESULTS: The LPS-treated group exhibited a remarkable increase in the extent of the pulmonary injury, oxidative stress indicator secretion, inflammatory molecule release, and inflammatory mediator production and an increase in the inhibitor of kappa B-α levels relative to the Sham group. The LYRM03 (5 and 10 mg/kg)-treated groups exhibited a remarkable decrease relative to the LPS group. In addition, treatment with LYRM03 (20 mg/kg) powerfully limited the extent of the injury and demonstrated anti-inflammatory actions. CONCLUSIONS: The results of this investigation indicated that treatment with LYRM03 plays a role in lung defense by inhibiting the NF-κB/MyD88/TLR4 pathway.

Influence of Dietary Copper Methionine Concentrations on Growth Performance, Digestibility of Nutrients, Serum Lipid Profiles, and Immune Defenses in Broilers.

A 42-day experiment was conducted to evaluate the influence of dietary copper (Cu) concentrations on growth performance, nutrient digestibility, and serum parameters in broilers aged from 1 to 42 days. Five hundred forty 1-day-old broilers were randomly assigned into 1 of the following 6 dietary treatments: (1) control (basal diet without supplemental Cu), (2) 15 mg/kg supplemental Cu (Cu15), (3) 30 mg/kg supplemental Cu (Cu30), (4) 60 mg/kg supplemental Cu (Cu60), (5) 120 mg/kg supplemental Cu (Cu120), and (6) 240 mg/kg supplemental Cu (Cu240), Cu as copper methionine. A 4-day metabolism trial was conducted during the last week of the experiment feeding. The results showed that dietary Cu supplementation increased the average daily gain and the average daily feed intake (P < 0.01). The feed gain ratio, however, was not affected by dietary Cu (P > 0.10). Additionally, dietary Cu supplementation increased the digestibility of fat and energy (P < 0.05). The concentration of serum cholesterol, triglycerides, and high-density lipoprotein cholesterol decreased with dietary Cu supplementation (P < 0.05). The activities of serum Cu-Zn superoxide dismutase (P < 0.05), glutathione peroxidase (P < 0.05), and ceruloplasmin (P = 0.09), on the contrary, were increased by Cu addition. For immune indexes, dietary Cu supplementation increased serum IgA and IgM (P < 0.05). In addition, the activities of serum ALT increased with increasing dietary Cu supplementation (P < 0.05). In conclusion, our data suggest that Cu supplementation can increase fat digestibility and promote growth. Additionally, dietary Cu supplementation can reduce serum cholesterol and enhance antioxidant capacity in broilers.

RNAi-mediated knockdown of MTNR1B without disrupting the effects of melatonin on apoptosis and cell cycle in bovine granulose cells.

Melatonin is well known as a powerful free radical scavenger and exhibits the ability to prevent cell apoptosis. In the present study, we investigated the role of melatonin and its receptor MTNR1B in regulating the function of bovine granulosa cells (GCs) and hypothesized the involvement of MTNR1B in mediating the effect of melatonin on GCs. Our results showed that MTNR1B knockdown significantly promoted GCs apoptosis but did not affect the cell cycle. These results were further verified by increasing the expression of pro-apoptosis genes (BAX and CASP3), decreasing expression of the anti-apoptosis genes (BCL2 and BCL-XL) and anti-oxidant genes (SOD1 and GPX4) without affecting cell cycle factors (CCND1, CCNE1 and CDKN1A) and TP53. In addition, MTNR1B knockdown did not disrupt the effects of melatonin in suppressing the GCs apoptosis or blocking the cell cycle. Moreover, MTNR1B knockdown did not affect the role of melatonin in increasing BCL2, BCL-XL, and CDKN1A expression, or decreasing BAX, CASP3, TP53, CCND1 and CCNE1 expression. The expression of MTNR1A was upregulated after MTNR1B knockdown, and melatonin promoted MTNR1A expression with or without MTNR1B knockdown. However, despite melatonin supplementation, the expression of SOD1 and GPX4 was still suppressed after MTNR1B knockdown. In conclusion, these findings indicate that melatonin and MTNR1B are involved in BCL2 family and CASP3-dependent apoptotic pathways in bovine GCs. MTNR1A and MTNR1B may coordinate the work of medicating the appropriate melatonin responses to GCs.

Elucidation of the Mechanisms of Long-Distance mRNA Movement in a Nicotiana benthamiana/Tomato Heterograft System.

Recent heterograft analyses showed that large-scale messenger RNA (mRNA) movement takes place in the phloem, but the number of mobile transcripts reported varies widely. However, our knowledge of the mechanisms underlying large-scale mRNA movement remains limited. In this study, using a Nicotiana benthamiana/tomato (Solanum lycopersicum) heterograft system and a transgenic approach involving potato (Solanum tuberosum), we found that: (1) the overall mRNA abundance in the leaf is not a good indicator of transcript mobility to the root; (2) increasing the expression levels of nonmobile mRNAs in the companion cells does not promote their mobility; (3) mobile mRNAs undergo degradation during their movement; and (4) some mRNAs arriving in roots move back to shoots. These results indicate that mRNA movement has both regulated and unregulated components. The cellular origins of mobile mRNAs may differ between herbaceous and woody species. Taken together, these findings suggest that the long-distance movement of mRNAs is a complex process and that elucidating the physiological roles associated with this movement is challenging but remains an important task for future research.

Effects of a Chimeric Lysin against Planktonic and Sessile Enterococcus faecalis Hint at Potential Application in Endodontic Therapy.

Enterococcus faecalis is a commensal opportunistic pathogen found in the intestine, mouth, and vaginal tract of humans. As an invasive pathogen in the oral cavity, E. faecalis is one of the leading causes of periapical endodontic lesions. However, due to the strong biofilm-forming capacity and tolerance of E. faecalis to conventional antibiotics and treatments, limited therapeutic options are available. In the present study, we investigated the activity of ClyR, a chimeric lysin with extended streptococcal lytic spectrum, against planktonic and sessile E. faecalis cells in vitro and in an ex vivo dental model. Our results showed that ClyR has robust and rapid lytic activity against multiple E. faecalis strains, killing >90% planktonic cells within 1 min at a concentration of 50 µg/mL. The biochemical experiments combined with microscopy analysis revealed that ClyR degrades E. faecalis biofilm with high efficacy in a dose-dependent manner, reducing the survival rate to 90% viable bacteria within biofilms at a low dose of 50 µg/mL, which is much better than ampicillin and similar to calcium hydroxide, the extensively used routine intracanal medicament in the treatment of endodontics and dental traumatology. The robust activity of ClyR against both planktonic and sessile E. faecalis suggests the potential of ClyR in treating endodontic infections caused by E. faecalis.

Effect of interaction of epigallocatechin gallate and flavonols on color alteration of simulative green tea infusion after thermal treatment.

Epigallocatechin gallate (EGCG) and flavonols are important phenolic compounds in green tea. These compounds are sensitive to thermal condition and their structural alteration results in making browning the green tea infusion. This study aimed to research the interaction between EGCG and flavonols during thermal infusion. EGCG and flavonols model solutions were prepared based on concentration in green tea infusion, and their colors appearance and attributes were analyzed in 10 h by thermal treatment. Results showed that kaempferol, quercetin, and myricetin accelerated the oxidation of EGCG and made the model solution browning. HPLC analysis revealed there was an obvious shift of a broaden peak in the mixed model solution of EGCG and flavonols after thermal treatment. This broaden peak was further purified on HPLC and solid phase extraction methods to yield colored complexes. The complexes showed the maximal absorption around 424, 442, and 482 nm. MS/MS analysis revealed that the complexes possessed of three components those were consisted of the interaction between EGCG and myricetin. These results indicated that the interaction between EGCG and flavonols might form complexes during thermal treatment, The complexes were contributed to make green tea infusion browning.

The effects of melatonin on bovine uniparental embryos development in vitro and the hormone secretion of COCs.

Melatonin is a unique multifunctional molecule that mediates reproductive functions in animals. In this study, we investigated the effects of melatonin on bovine parthenogenetic and androgenetic embryonic development, oocyte maturation, the reactive oxygen species (ROS) levels in parthenogenetic and androgenetic embryos and cumulus-oocyte complexes (COCs) hormone secretion with melatonin supplementation at four concentrations (0, 10, 20, and 30 pmol/mL), respectively. The results showed that melatonin significantly promoted the rates of bovine parthenogenetic and androgenetic embryonic cleavage and morula and blastocysts development (P < 0.05). The rate of cleavage was higher in the androgenetic embryo than that in the parthenogenetic embryo. Compared with the parthenogenetic embryos, the androgenetic embryos had a poor developmental competence from morula to blastocyst stage. Moreover, the levels of ROS were significantly lower in the parthenogenetic and androgenetic embryoes with melatonin-treated group than that of the control group (P < 0.05). Melatonin supplemented significantly increased the maturation rate of oocyte in vitro (P < 0.05). More importantly, melatonin significantly promoted the secretion of progesterone and estradiol by COCs (P < 0.05). To reveal the regulatory mechanism of melatonin on steroids synthesis, we found that steroidogenic genes (CYP11A1, CYP19A1 and StAR) were upregulated, suggesting that melatonin regulated estradiol and progesterone secretion through mediating the expression of steroidogenic genes (CYP11A1, CYP19A1 and StAR). In addition, MT1 and MT2 were identified in bovine early parthenogenetic and androgenetic embryos using western blot. It could be concluded that melatonin had beneficial effects on bovine oocyte in vitro maturation, COC hormone secretion, early development of subsequent parthenogenetic and androgenetic embryos. It is inferred that melatonin could be used to enhance the efficiency of in vitro developed embryos.

[Effect of Wenyang Decoction on the Differentiation of CD34+ Progenitor Cells in Occupational Asthma Model Rats].

OBJECTIVE: To study the effect of Wenyang Decoction (WD) on the differentiation of CD34+ progenitor cells of occupational asthma (OA) model rats. METHODS: Fifty healthy male SD rats were randomly divided into five groups, i.e., the model group, the blank control group,the WD group,the Western medicine group,the combined group, 10 in each group. Prednisone suspension (10 mg/kg) was administered to rats in the Western medicine group by gastrogavage. WD (20 g/kg) was administered to rats in the WD group by gastrogavage. Prednisone suspension plus WD was administered to rats in the combined group by gastrogavage. Normal saline was administered to rats in the model group and the blank control group by gastrogavage. The general condition of rats was observed. Expression levels of peripheral blood IL-5 and eotaxin, eosinophils (EOS), CD34+, CC chemokine receptor 3 (CCR3+) in bone marrow suspension were detected by ELISA, Wirght-Giemsa, and flow cytometry, respectively. RESULTS: Compared with the blank control group,expression levels of IL-5 and eotaxin in peripheral blood were significantly higher (P < 0.01), and the count of EOS and CD34+ cells, as well as CD34+ /CCR3+ significantly increased (P < 0.01) in the model group. Compared with the model group, expression levels of IL-5 and eotaxin, the count of EOS, CD34+ cells, CD34+ / CCR3+ were lowered in three treated groups (P < 0.01). Compared with the Western medicine group, the count of EOS and CD34+ / CCR3+ decreased in the combined group (P < 0.01). The count of EOS was significantly lower in the combined group than in the WD group (P < 0.01). CONCLUSION: WD could reduce levels of in vivo inflammatory factors, and restrain the differentiation and recruitment of EOS,thereby alleviating the differentiation of CD34 progenitor cells to EOS.

MALDI imaging for the localization of saponins in root tissues and rapid differentiation of three Panax herbs.

The roots of Panax genus with ginseng saponins as bioactive ingredients have been widely used as herbal medicines and food additives. Panax ginseng, Panax quinquefolius, and Panax notoginseng are three major commercial species in Panax genus, with similar morphological appearance but different pharmacological functions. Various methods have been developed and applied for the differentiation of these species. In this work, MALDI-TOF-MS imaging (MSI) was employed for the localization of saponins in root tissues and for the rapid differentiation of the three Panax species for the first time. After a simple sample preparation, MALDI-TOF-MSI analysis of root tissue allowed the detection of 51 saponins. Localization of saponins in the tissue was mapped in ion images, which were obviously related to botanical structure. The localization modes varied with Panax species, providing valuable information for the discrimination of ginseng species. Principal component analysis (PCA) of data collected from areas with abundant saponins based on ion images was applied for the differentiation. Nine characteristic saponin peaks were identified from the PCA analysis. The MALDI-TOF-MSI together with area-specific data analysis provided high potential for the rapid differentiation of Panax herbs.

Rosiglitazone Promotes Bone Marrow Adipogenesis to Impair Myelopoiesis under Stress.

OBJECTIVE: The therapeutic use of thiazolidinediones (TZDs) causes unwanted hematological side effects, although the underlying mechanisms of these effects are poorly understood. This study tests the hypothesis that rosiglitazone impairs the maintenance and differentiation of hematopoietic stem/progenitor cells, which ultimately leads to hematological abnormalities. METHODS: Mice were fed a rosiglitazone-supplemented diet or a normal diet for 6 weeks. To induce hematopoietic stress, all mice were injected once with 250 mg/kg 5-fluorouracil (5-Fu) intraperitoneally. Next, hematopoietic recovery, hematopoietic stem/progenitor cells (HSPCs) subsets, and myeloid differentiation after 5-Fu treatment were evaluated. The adipogenesis induced by rosiglitazone was assessed by histopathology and oil red O staining. The effect of adipocytes on HSPCs was studied with an in vitro co-culture system. RESULTS: Rosiglitazone significantly enhanced bone marrow adipogenesis and delayed hematopoietic recovery after 5-Fu treatment. Moreover, rosiglitazone inhibited proliferation of a granulocyte/monocyte progenitor (GMP) cell population and granulocyte/macrophage colony-stimulating factor (GM-CSF) colonies, although the proliferation and mobilization of Lin-c-kit+Sca-1+ cells (LSK) was maintained following hematopoietic stress. These effects could be partially reversed by the selective PPARγ antagonist BADGE. Finally, we demonstrated in a co-culture system that differentiated adipocytes actively suppressed the myeloid differentiation of HSPCs. CONCLUSION: Taken together, our results demonstrate that rosiglitazone inhibits myeloid differentiation of HSPCs after stress partially by inducing bone marrow adipogenesis. Targeting the bone marrow microenvironment might be one mechanism by which rosiglitazone impairs stress-induced hematopoiesis.