Modeling human pregastrulation development by 3D culture of blastoids generated from primed-to-naïve transitioning intermediates

Human pluripotent stem cells provide an inexhaustible model to study human embryogenesis in vitro. Recent studies have provided diverse models to generate human blastoids by self-organization of different pluripotent stem cells or somatic reprogramming intermediates. However, whether blastoids can be generated from other cell types or whether they can recapitulate postimplantation development in vitro is unknown. Here, we develop a strategy to generate human blastoids from heterogeneous intermediates with epiblast, trophectoderm, and primitive endoderm signatures of the primed-to-naïve conversion process, which resemble natural blastocysts in morphological architecture, composition of cell lineages, transcriptome, and lineage differentiation potential. In addition, these blastoids reflect many features of human peri-implantation and pregastrulation development when further cultured in an in vitro 3D culture system. In summary, our study provides an alternative strategy to generate human blastoids and offers insights into human early embryogenesis by modeling peri- and postimplantation development in vitro.

Association of recurrent upper respiratory tract infections with low production of oxygen intermediates in children

Abstract Objectives: Upper respiratory tract infections in children generally have significant morbidity and mortality. There is little data available about functional immaturity of the immune system and the child's susceptibility to infections at the beginning of their lives, thus, justifying a more specific immunological analysis. Method: Analysis of hemograms and innate and adaptive immune responses in 95 children between age 1 to 6 years with episodes of recurrent respiratory infections (test group n = 39) and without these episodes (control group n = 56) was carried out. The production of reactive oxygen intermediates by peripheral blood cells stimulated by phorbol myristate acetate was analyzed. Additionally, the number of B lymphocytes, auxiliary T lymphocytes, and cytotoxic cells was determined using flow cytometry. Results: Results from both groups did not show statistically significant differences in red blood cells, total leukocytes count, and the differential neutrophils, eosinophils, basophils, lymphocytes, and monocytes count. The analysis of the number of B lymphocytes, auxiliary T lymphocytes (LTCD4), and cytotoxic cells (LTCD8) also did not show any difference between both groups. However, the production of radical oxygen intermediates was significantly reduced in the test group as compared to the control group (p < 0.05). Conclusions: There was no difference in the analysis of hemograms, leukograms, or the number of lymphocytes, LTCD4, LTCD8, or LTCD19. The reduced production of oxygen intermediates in the affected group suggests that these children's microbicide capacity is compromised, which may be related to their recurrent respiratory infections.

Research progress on synthesis of intermediates in derivatization of β-elemene / 中草药

β-Elemene is the main active ingredient of elemene, a broad-spectrum anti-cancer botanical drug, which is also the main isomer of elemene's four isomers. The synthesis of some key intermediates were generalized and summarized during the derivatization of β-elemene, wherein the preparation methods, reaction conditions, yields, characterization data of these intermediates were listed. The purpose of this paper is to facilitate people in understanding and grasping the most optimal synthesis of these intermediates up to date, to inspire people pursuing further synthesis improvement, and to stimulate generation of new ideas for future derivatization of β-elemene.

Correlation Analysis of HPLC Fingerprint of Pieces, Standard Decoction, Intermediates and Dispensing Granules of Rehmanniae Radix Praeparata / 中国实验方剂学杂志

Objective::To establish HPLC fingerprint spectra of the pieces, standard decoction, intermediates, dispensing granules of Rehmanniae Radix Praeparata, and assess the quality correlation among them, then to evaluate the scientificity and rationality of preparation process based on the yields of dry extract and the transfer rate of acteoside. Method::Fingerprints of several batches of the pieces, standard decoction, intermediates and dispensing granules of Rehmanniae Radix Praeparata were detected by HPLC, and the content of acteoside was determined according to the method of ChP 2015.The fingerprint chromatographic separation was carried out on Phenomenex Luna 100A C18(2) chromatographic column (4.6 mm×250 mm, 5 μm). The mobile phase was acetonitrile-0.1% phosphoric acid for gradient elution, with a flow rate of 1 mL·min-1, and the detection wavelength was 330 nm. At the same time, the correlation analysis of quality transmission during the preparation of dispensing granules was carried out based on the yields of dry extract and the transfer rates of acteoside. Result::The contents of acteoside pieces, standard decoction and intermediates were basically consistent. The yield of dry extracts of intermediates and dispensing granules, and the transmission rate of acteoside were all within the range of standard decoction, and basically consistent with standard decoction. There were 7 common peaks in all fingerprint spectra of 17 batches of pieces, 17 batches of standard decoction, 10 intermediates and 10 dispensing granules of Rehmanniae Radix Praeparata, with a good correlation. The 13 main chromatographic peaks in the dispensing granules were identified by UPLC-Q-TOF-MS analysis, and 4 of the 7 fingerprint common peaks were identified as 5-hydroxymethyl furfural, acteoside, isoacteoside and martynoside. Conclusion::The main chemical constituents of Rehmanniae Radix Praeparata pieces, standard decoction, intermediates and dispensing granules are basically identical. The established HPLC fingerprint method can be used for the quality control of preparation process of Rehmanniae Radix Praeparata dispensing granules.

Synthesis of (S)-N-Boc-3-hydroxyadamantylglycine a key intermediate of Saxagliptin: Research advances / 国际药学研究杂志

Saxagliptin, a novel antihyperglycemic agent belonging to dipeptidyl peptidase-4 (DPP-4) inhibitor, was developed by Bristol-Myers Squibb and AstraZeneca. It has the merit of obvious hypoglycemic activity, less side effect, good treatment compliance and remarkably high safety, and is widely applied to treatment of type 2 diabetes. In this paper, we review the synthetic methods of the key intermediate (S)-N-Boc-3-hydroxyadamantylglycine (compound 3) as well as its two important compounds 3-hydroxy-1-acetyladamantane (1) and 2-(3-hydroxy-1-adamant-yl)-2-oxoacetic acid (2) and briefly discuss their advantages and disadvantages.

Synthesis of(S)-N-Boc-3-hydroxyadamantylglycine a key intermediate of saxagliptin:research advances / 国际药学研究杂志

Saxagliptin,a novel antihyperglycemic agent belonging to dipeptidyl peptidase-4(DPP-4)inhibitor,was devel?oped by Bristol-Myers Squibb and AstraZeneca. It has the merit of obvious hypoglycemic activity,less side effect,good treatment com?pliance and remarkably high safety,and is widely applied to treatment of type 2 diabetes. In this paper,we review the synthetic meth?ods of the key intermediate(S)-N-Boc-3-hydroxyadamantylglycine(compound 3)as well as its two important compounds 3-hydroxy-1-acetyladamantane(1)and 2-(3-hydroxy-1-adamant-yl)-2-oxoacetic acid(2)and briefly discuss their advantages and disadvantages.

Two step process for diketo-reduction catalyzed by diketoreductase / 中国药科大学学报

Diketoreductase can catalyze a double reduction of β,δ-diketo ester to corresponding dihydroxv product with enantiomeric excess(ee)and diastereomeric excess(de)both greater than 99.5%.In order to explore the catalytic mechanism of this unique enzyme,the present study investigated the diketone reduction process and reaction characteristics by diketoreductase.We found that two mono-hydroxy intermediates were produced during the diketone reduction and its reverse reaction.The two intermediates were further separated by C18 column chromatography and structurally confirmed by chiral HPLC with authentic standards.Two mono-hydroxy intermediates could be served as the substrates for the reduction by diketoreductase with different reaction velocities.The formation and disappearance of intermediates were largely affected by temperature and substrate concentration.In addition,steady state kinetics with the two intermediates showed different reaction rates in their disappearance.Collectively,the results indicate that the diketone reduction undergoes a two-step process with the formation of both intermediates,but the disappearance rates for the two intermediates are slightly different.

Egr-1 promoter regulating effect on granulocyte-macrophage colony-stimulating factor (GM-CSF) gene expression induced by doxorubicin and ionizing radiation / 中华放射医学与防护杂志

Objective To explore the regulating effects of Egr-1 promoter activated by ionizing radiation (IR) and doxorubicin (ADM) on the expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) genes. Methods The human GM-CSF cDNA and enhanced green fluorescent protein (EGFP) cDNA were linked together with IRES(internal ribosome entry site) and then inserted into the expression vector pCIneo under control of the Egr-1 promoter(Egr-EG). The vector was transferred into human bone marrow stromal cell line HFCL by liposome transfection. And the cells were exposure to ADM and IR. The activity of EGFP in HFCL/EG cells were detected by FACS. The effect of N-acetylcysteine on the expression of EGFP following exposure to ADM and IR was examined. The amounts of GM-CSF in HFCL/EG after chemotherapy or radiation were measured with ELISA. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM derived from cord blood were also studied. RT-PCR analysis for the expression of GM-CSF mRNA in HFCL/EG after exposure to ADM or IR. Results The percentage of EGFP+ HFCL/EG cells exposed to ADM and IR was increased compared with non-treatment group (1.2 % and 15.2 % vs 18.2 %, t = 5.11, P < 0.01). The levels of secreted GM-CSF in HFCL/EG cells exposed to ADM and IR was increased (P < 0.01), but no difference between ADM group and IR group (P 0.05). The expression of EGFP by HFCL/EG treated with ADM and IR was significantly decreased by N-acetylcysteine. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM in ADM group and IR group were significantly higher than that in HFCL group and non-treatment group. However, The CFU-GM count of IR group was higher than that of ADM group. The expression of GM-CSF mRNA in HFCL/EG cells exposed to ADM and IR was significantly increased(t = 4.37, P < 0.01). Conclusions GM-CSF gene expression regulated by Egr-1 promoter induced by ADM and IR could help the recovery from hematopoietic injury.

The impact of glucose-6-phosphate dehydrogenase deficiency in Chinese on neutrophil / 医学研究生学报

Objective: The aim of this study is to evaluate the impact of glucose-6- phosphate dehydrogenase deficiency in Chinese on the intracellular glucose-6-phosphate dehydrogenase activity and reactive oxygen intermediates production of peripheral neutrophil. Methods: Glucose-6-phosphate dehydrogenase, its mRNA expression and reactive oxygen intermediates production(NBT test) in silent or stimulated neutrophil from health controls and patients with glucose-6-phosphate dehydrogenase deficiency were analysized. Results: Compared with health controls, the activities of glucose-6-phosphate dehydrogenase of peripheral neutrophil from patients with glucose-6 -phosphate dehydrogenase deficiency was decreased, and its mRNA expression increased. Reactive oxygen intermediate production increased(no statistical signification) in silence but decreased following stimulation by LPS in neutrophil from normal individuals than that from patients. Conclusion: Decreased activity of G-6-PD and production of reactive oxygen intermediates following stimulation in neutrophil could be caused by G-6-PD deficiency in Chinese.

Isolation and identification of Murine Cerebral Microglial Cells

Even though the brain has been considered to be an immunologically privileged organ, recent reports showed that certain cells of the brain may be involved in immunological process in the brain. For example, some cells of the brain can present antigen to T-lymphocytes, to express class II major histocompativility antigen, and secrete interleukin-1 and -3 molecules. In addition, they are capable to phagocytose particles and possess receptors for the Fc portion on IgG. In this study, the authors tried to isolate the microglial cells from new born mice and characterize them. The isolated cells could produce such reactive oxygen intermediates(ROIs) an superoxide and hydrogen peroxide that were measured by luminometer after amplification by lucigenin and luminol respectively and could secrete reactive nitrogen intermediates(RNIs), when the cells were incubated with r-IFN plus LPS. The cells could also ingest fluorescent particles and raise intracellular calcium after stimulation with agonists when measured by flow cytometer. Our data showed that the microglial cells of the brain may belong to a member of mononuclear phagocytic system(MPS) of the body that are responsible for the host defence against invading microorganisms.

Advances in Microbial Transformation of Phytosterol into Steroid Medicine Intermediates / 微生物学通报

Microbial selective side-chain degradation of phytosterol,which can obtain the steroid medicine intermediate compounds-androst-4-ene-3,17-dione (4-AD) and androsta-1,4-diene-3,17-dione (ADD),has an important meaning to pharmacy. There is no systematic literature concerned in existence. Its mechnism,approaches,influencing factors and so on over these years were fully reviewed in the paper. The trend of development in the area is expanded.