Pulmonary blastoma treatment response to anti-PD-1 therapy: a rare case report and literature review.

Pulmonary blastoma (PB) is a rare and invasive malignancy of the lungs with a poor prognosis. Although the mainstay treatment of PB is surgery, and radiotherapy and chemotherapy have been reported, no standard therapy exists for patients inoperable in advanced stages. Moreover, little is known about driver mutation status and immunotherapy efficacy. This paper presents a male patient diagnosed with classic biphasic PB using CT-guided lung biopsy pathology and immunohistochemistry. The patient's symptoms included cough, chest pain, shortness of breath, hemoptysis, and hypodynamia. The primary focus of this paper is to discuss the impact of anti-PD-1 immunotherapy on PB. The patient experienced progression-free survival (PFS) of over 27 months following sintilimab second-line anti-PD-1 therapy. The patient has currently survived for nearly 40 months with a satisfactory quality of life.

Serendipita indica promotes P acquisition and growth in tea seedlings under P deficit conditions by increasing cytokinins and indoleacetic acid and phosphate transporter gene expression.

The culturable endophytic fungus Serendipita indica has many beneficial effects on plants, but whether and how it affects physiological activities and phosphorus (P) acquisition of tea seedlings at low P levels is unclear. The objective of this study was to analyze the effects of inoculation with S. indica on growth, gas exchange, chlorophyll fluorescence, auxins, cytokinins, P levels, and expressions of two phosphate transporter (PT) genes in leaves of tea (Camellia sinensis L. cv. Fudingdabaicha) seedlings grown at 0.5 µM (P0.5) and 50 µM (P50) P levels. Sixteen weeks after the inoculation, S. indica colonized roots of tea seedlings, with root fungal colonization rates reaching 62.18% and 81.34% at P0.5 and P50 levels, respectively. Although plant growth behavior, leaf gas exchange, chlorophyll values, nitrogen balance index, and chlorophyll fluorescence parameters of tea seedlings were suppressed at P0.5 versus P50 levels, inoculation of S. indica mitigated the negative effects to some extent, along with more prominent promotion at P0.5 levels. S. indica inoculation significantly increased leaf P and indoleacetic acid concentrations at P0.5 and P50 levels and leaf isopentenyladenine, dihydrozeatin, and transzeatin concentrations at P0.5 levels, coupled with the reduction of indolebutyric acid at P50 levels. Inoculation of S. indica up-regulated the relative expression of leaf CsPT1 at P0.5 and P50 levels and CsPT4 at P0.5 levels. It is concluded that S. indica promoted P acquisition and growth in tea seedlings under P deficit conditions by increasing cytokinins and indoleacetic acid and CsPT1 and CsPT4 expression.

Prognostic implications of left ventricular torsion measured by feature-tracking cardiac magnetic resonance in patients with ST-elevation myocardial infarction.

AIMS: The prognostic implication of left ventricular (LV) torsion on ST-elevation myocardial infarction (STEMI) is unclear. METHODS AND RESULTS: We analysed cardiovascular magnetic resonance (CMR) findings of 420 patients from a registry study (NCT03768453). These patients received CMR examination within 1 week after timely primary percutaneous coronary intervention. LV torsion and other CMR indexes were measured. Compared with healthy control subjects, STEMI significantly decreased patients' LV torsion (1.04 vs. 1.63°/cm, P < 0.001). During follow-up (median, 52 months), the reduction of LV torsion was greater in patients with than without composite major adverse cardiac and cerebrovascular events (MACCEs, 0.79 vs. 1.08°/cm, P < 0.001). The risk of MACCEs would increase to 1.125- or 1.092-fold, and the risk of 1-year LV remodelling would increase to 1.110- or 1.082-fold for every 0.1°/cm reduction in LV torsion after adjustment for clinical or CMR parameters respectively. When divided dichotomously, patients with LV torsion≤ 0.802°/cm had significantly higher risk of MACCEs (40.2 vs. 12.3%, P < 0.001) and more remarkable LV remodelling (46.1 vs. 11.9%, P < 0.001) than patients with better LV torsion. The addition of LV torsion to conventional prognostic factors such as the LV ejection fraction and infarction size led to a better risk classification model of patients for both MACCEs and LV remodelling. Finally, tobacco use, worse post-PCI flow, and greater microvascular obstruction size were presumptive risk factors for reduced LV torsion. CONCLUSION: LV torsion measured by CMR is closely associated with the prognosis of STEMI and would be a promising indicator to improve patients' risk stratification. CLINICAL TRIAL REGISTRATION: Clinicaltrials.gov, NCT03768453.

Carotenoids Biosynthesis, Accumulation, and Applications of a Model Microalga Euglenagracilis.

The carotenoids, including lycopene, lutein, astaxanthin, and zeaxanthin belong to the isoprenoids, whose basic structure is made up of eight isoprene units, resulting in a C40 backbone, though some of them are only trace components in Euglena. They are essential to all photosynthetic organisms due to their superior photoprotective and antioxidant properties. Their dietary functions decrease the risk of breast, cervical, vaginal, and colorectal cancers and cardiovascular and eye diseases. Antioxidant functions of carotenoids are based on mechanisms such as quenching free radicals, mitigating damage from reactive oxidant species, and hindering lipid peroxidation. With the development of carotenoid studies, their distribution, functions, and composition have been identified in microalgae and higher plants. Although bleached or achlorophyllous mutants of Euglena were among the earliest carotenoid-related microalgae under investigation, current knowledge on the composition and biosynthesis of these compounds in Euglena is still elusive. This review aims to overview what is known about carotenoid metabolism in Euglena, focusing on the carotenoid distribution and structure, biosynthesis pathway, and accumulation in Euglena strains and mutants under environmental stresses and different culture conditions. Moreover, we also summarize the potential applications in therapy preventing carcinogenesis, cosmetic industries, food industries, and animal feed.

Concomitant Pulmonary Tuberculosis Impair Survival in Advanced Epidermal Growth Factor Receptor (EGFR) Mutant Lung Adenocarcinoma Patients Receiving EGFR-Tyrosine Kinase Inhibitor.

OBJECTIVE: Limited studies have clearly demonstrated the effect of EGFR-TKI in the treatment of EGFR mutant NSCLC patients with underlying pulmonary disease, like pulmonary tuberculosis (PTB). Here, we conducted the study to evaluate the impact of PTB on survival of Chinese EGFR mutant lung adenocarcinoma (LUAD) patients that underwent EGFR-TKI treatment. METHODS: Clinicopathologic data of 1448 LUAD patients harboring EGFR mutations from the Guangzhou Chest Hospital between 2017 and 2019 were reviewed retrospectively. Patients receiving EGFR-TKI treatment were divided into PTB and non-PTB groups. The differences in response to EGFR-TKIs and survival between the two groups were assessed. RESULTS: After EGFR-TKIs treatment, the objective response rate (58.14% vs 47.62%) as well as disease control rate (97.67% vs 85.71%) were higher in the non-PTB group than in the PTB group, but there was no statistical difference. In the survival analysis, both the median progression-free survival (7.47 months vs 11.77 months, p = 0.038) and the overall survival (13.00 months vs 20.00 months, p = 0.001) were significantly shorter in the PTB group than in the non-PTB group. Furthermore, for patients with 19Del mutation, or metastases sites less than 3, or using first-line EGFR-TKI, EGFR-TKIs treatment significantly prolonged the median PFS and OS in patients without PTB. CONCLUSION: LUAD patients with concomitant PTB have a poor response to EGFR-TKI treatment, especially in terms of survival outcome.

Impact of Concomitant Impairments of the Left and Right Ventricular Myocardial Strain on the Prognoses of Patients With ST-Elevation Myocardial Infarction.

Background: The impact of concomitant impairments of left and right ventricular (LV and RV) strain on the long-term prognosis of acute ST-elevation myocardial infarction (STEMI) is not clear. Methods: We analyzed CMR images and followed up 420 first STEMI patients from the EARLY Assessment of MYOcardial Tissue Characteristics by CMR in STEMI (EARLY-MYO-CMR) registry (NCT03768453). These patients received timely primary percutaneous coronary intervention (PCI) within 12 h and CMR examination within 1 week (median, 5 days; range, 2-7 days) after infarction. Global longitudinal strain (GLS), global radial strain (GRS), and global circumferential strain (GCS) of both ventricles were measured based on CMR cine images. Conventional CMR indexes were also assessed. Primary clinical outcome was composite major adverse cardiac and cerebrovascular events (MACCEs) including cardiovascular death, re-infarction, re-hospitalization for heart failure and stroke. In addition, CMR data from 40 people without apparent heart disease were used as control group. Results: Compared to controls, both LV and RV strains were remarkably reduced in STEMI patients. During follow-up (median: 52 months, interquartile range: 29-68 months), 80 patients experienced major adverse cardiac and cerebrovascular events (MACCEs) including cardiovascular death, re-infarction, heart failure, and stroke. LV-GCS > -11.20% was an independent predictor of MACCEs (P < 0.001). RV-GRS was the only RV strain index that could effectively predict the risk of MACCEs (AUC = 0.604, 95% CI [0.533, 0.674], P = 0.004). Patient with RV-GRS ≤ 38.79% experienced more MACCEs than those with preserved RV-GRS (log rank P < 0.001). Moreover, patients with the concomitant decrease of LV-GCS and RV-GRS were more likely to experience MACCEs than patients with decreased LV-GCS alone (log rank P = 0.010). RV-GRS was incremental to LV-GCS for the predictive power of MACCEs (continuous NRI: 0.327; 95% CI: 0.095-0.558; P = 0.006). Finally, tobacco use (P = 0.003), right coronary artery involvement (P = 0.002), and LV-GCS > -11.20% (P = 0.012) was correlated with lower RV-GRS. Conclusions: The concomitant decrease of LV and RV strain is associated with a worse long-term prognosis than impaired LV strain alone. Combination assessment of both LV and RV strain indexes could improve risk stratification of patients with STEMI. Trial Registration: ClinicalTrials.gov, NCT03768453. Registered 7 December 2018 - Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT03768453.

Comparative Genomics, Evolutionary and Gene Regulatory Regions Analysis of Casein Gene Family in Bubalus bubalis.

Buffalo is a luxurious genetic resource with multiple utilities (as a dairy, draft, and meat animal) and economic significance in the tropical and subtropical regions of the globe. The excellent potential to survive and perform on marginal resources makes buffalo an important source for nutritious products, particularly milk and meat. This study was aimed to investigate the evolutionary relationship, physiochemical properties, and comparative genomic analysis of the casein gene family (CSN1S1, CSN2, CSN1S2, and CSN3) in river and swamp buffalo. Phylogenetic, gene structure, motif, and conserved domain analysis revealed the evolutionarily conserved nature of the casein genes in buffalo and other closely related species. Results indicated that casein proteins were unstable, hydrophilic, and thermostable, although αs1-CN, ß-CN, and κ-CN exhibited acidic properties except for αs2-CN, which behaved slightly basic. Comparative analysis of amino acid sequences revealed greater variation in the river buffalo breeds than the swamp buffalo indicating the possible role of these variations in the regulation of milk traits in buffalo. Furthermore, we identified lower transcription activators STATs and higher repressor site YY1 distribution in swamp buffalo, revealing its association with lower expression of casein genes that might subsequently affect milk production. The role of the main motifs in controlling the expression of casein genes necessitates the need for functional studies to evaluate the effect of these elements on the regulation of casein gene function in buffalo.

Tumor Mutational Burden and Genomic Alterations in Chinese Small Cell Lung Cancer Measured by Whole-Exome Sequencing.

PURPOSE: Studies on genetic alterations of the heterogenous small cell lung cancer (SCLC) are rare. We carried out the present study to clarify the genomic alterations and TMB levels of Chinese SCLC patients by whole-exome sequencing. MATERIALS AND METHODS: Whole-exome sequencing by next-generation sequencing technique was implemented on twenty SCLC samples. Significant somatic mutations and copy number variations were screened, followed by comparison with the data extracted from COSMIC. Besides, altered signaling pathways were examined in order to figure out actionable targets. RESULTS: A total of 8,062 nonsynonymous mutations were defined. The number of mutations for each case ranged from 98 to 864. As for base substitutions, a total of 15,817 substitutions were detected with C > A conversion which was correlated to smoking occupying 25.57%. The TMB values ranged from 2.51/Mb to 22.1/Mb with a median value of 9.95/Mb. RB1 was the most frequently mutated gene altered in 18 (90%) cases, followed by TP53 altered in 17 (85%) cases. Other commonly changed genes were PTEN, and RBL1, with frequencies of 55% and 50%, respectively. SOX2 significantly amplified in 6 (30%) cases and MYCN amplified in 1 (5%) patient. Notch signaling pathway and PI3K/AKT/mTOR signaling pathway were universally and significantly changed. Major genomic alterations were in consistency with data from COSMIC, but frequencies of less common mutations were different. CONCLUSION: TP53 and RB1 inactivations were universally detected in SCLC. The Notch and PI3K/AKT/mTOR signaling pathways were both significantly altered, implying potential actionable targets.

Characterization of porcine extraembryonic endoderm cells.

OBJECTIVES: To date, many efforts have been made to establish porcine embryonic stem (pES) cells without success. Extraembryonic endoderm (XEN) cells can self-renew and differentiate into the visceral endoderm and parietal endoderm. XEN cells are derived from the primitive endoderm of the inner cell mass of blastocysts and may be an intermediate state in cell reprogramming. MATERIALS AND METHODS: Porcine XEN cells (pXENCs) were generated from porcine pluripotent stem cells (pPSCs) and were characterized by RNA sequencing and immunofluorescence analyses. The developmental potential of pXENCs was investigated in chimeric mouse embryos. RESULTS: Porcine XEN cells derived from porcine pPSCs were successfully expanded in N2B27 medium supplemented with bFGF for least 30 passages. RNA sequencing and immunofluorescence analyses showed that pXENCs expressed the murine and canine XEN markers Gata6, Gata4, Sox17 and Pdgfra but not the pluripotent markers Oct4, Sox2 and TE marker Cdx2. Moreover, these cells contributed to the XEN when injected into four-cell stage mouse embryos. Supplementation with Chir99021 and SB431542 promoted the pluripotency of the pXENCs. CONCLUSIONS: We successfully derived pXENCs and showed that supplementation with Chir99021 and SB431542 confer them with pluripotency. Our results provide a new resource for investigating the reprogramming mechanism of porcine-induced pluripotent stem cells.

[Research progress on the asymmetric division in mammalian oocytes].

The mammalian oocyte maturation process consists of two consecutive asymmetric divisions, and produces three daughter cells of vastly different sizes: one larger egg cell and two smaller polar bodies. Asymmetric division is a typical feature of mammalian oocyte meiosis that results in a highly polar egg cell. The mitosis of the cell after fertilization exhibits restored symmetric division, but the polarity characteristics formed during meiosis of oocytes are preserved and affect the polarity of early embryos. In this review, we summarize the research progress on asymmetric division of mammalian oocytes in recent years, and mainly focus on the asymmetric division of cytoplasmic and the asymmetric division of nucleus, including the functions of chromosome and cytoskeleton in asymmetric division of mammalian oocytes, the redistribution of cytoplasmic organelles occurring in oocyte maturation, and chromosome nonrandom separation. We aim to demonstrate the main mechanism of asymmetry division in mammalian oocytes from both cellular and molecular levels.

Does energetic cost for leaf construction in Sonneratia change after introduce to another mangrove wetland and differ from native mangrove plants in South China?

Exotic species invasions are serious ecological problems. Leaf construction cost (CC) and growth traits of two Sonneratia (Sonneratia caseolaris and S. apetala) and four native species (Bruguiera gymnorrhiza, Kandelia obovata, Aegiceras corniculatum and Avicennia marina) in Hainan and Shenzhen mangrove wetlands were compared to evaluate invasive potentials of Sonneratia after introduced to Shenzhen, their new habitat. There were no significant differences in CC and growth traits between two wetlands, suggesting Sonneratia did not lose any advantage in the new habitat and were competitive in both wetlands. CC per unit mass (CCM), CC per unit area (CCA) and caloric values of Sonneratia were significantly lower than those of native mangrove species while specific leaf area (SLA) was just the opposite. CCM of S. caseolaris and S. apetala were 6.1% and 11.9% lower than those of natives, respectively. These findings indicated the invasive potential of Sonneratia in Shenzhen after their introduction.

Relationship of Soluble Grape-Derived Proteins to Condensed Tannin Extractability during Red Wine Fermentation.

In red winemaking, the extractability of condensed tannins (CT) can vary considerably even under identical fermentation conditions, and several explanations for this phenomenon have been proposed. Recent work has demonstrated that grape pathogenesis-related proteins (PRPs) may limit retention of CT added to finished wines, but their relevance to CT extractability has not been evaluated. In this work, Vitis vinifera and interspecific hybrids (Vitis ssp.) from both hot and cool climates were vinified under small-scale, controlled conditions. The final CT concentration in wine was well modeled from initial grape tannin and juice protein concentrations using the Freundlich equation (r2= 0.686). In follow-up experiments, separation and pretreatment of juice by bentonite, heating, freezing, or exogenous tannin addition reduced protein concentrations in juices from two grape varieties. The bentonite treatment also led to greater wine CT for one of the varieties, indicating that prefermentation removal of grape protein may be a viable approach to increasing wine CT.

Are Photosynthetic Characteristics and Energetic Cost Important Invasive Traits for Alien Sonneratia Species in South China?

A higher photosynthesis and lower energetic cost are recognized as important characteristics for invasive species, but whether these traits are also important for the ability of alien mangrove species to become invasive has seldom been reported. A microcosm study was conducted to compare the photosynthetic characteristics, energetic cost indices and other growth traits between two alien species (Sonneratia apetala and S. caseolaris) and four native mangrove species over four seasons in a subtropical mangrove nature reserve in Shenzhen, South China. The aim of the study was to evaluate the invasive potential of Sonneratia based on these physiological responses. The annual average net photosynthetic rate (Pn), stomatal conductance (Gs) and total carbon assimilation per unit leaf area (Atotal) of the two alien Sonneratia species were significantly higher than the values of the native mangroves. In contrast, the opposite results were obtained for the leaf construction cost (CC) per unit dry mass (CCM) and CC per unit area (CCA) values. The higher Atotal and lower CC values resulted in a 72% higher photosynthetic energy-use efficiency (PEUE) for Sonneratia compared to native mangroves, leading to a higher relative growth rate (RGR) of the biomass and height of Sonneratia with the respective values being 51% and 119% higher than those of the native species. Higher photosynthetic indices for Sonneratia compared to native species were found in all seasons except winter, whereas lower CC values were found in all four seasons. The present findings reveal that alien Sonneratia species may adapt well and become invasive in subtropical mangrove wetlands in Shenzhen due to their higher photosynthetic characteristics coupled with lower costs in energy use, leading to a higher PEUE. The comparison of these physiological responses between S. apetala and S. caseolaris reveal that the former species is more invasive than the latter one, thus requiring more attention in future.

Characterization and comparative analyses of transcriptomes for in vivo and in vitro produced peri-implantation conceptuses and endometria from sheep.

An increasing number of reports indicate that in vitro fertilization (IVF) is highly associated with long­term side effects on embryonic and postnatal development, and can sometimes result in embryonic implant failure. While high­throughput gene expression analysis has been used to explore the mechanisms underlying IVF-induced side effects on embryonic development, little is known about the effects of IVF on conceptus-endometrial interactions during the peri-implantation period. Using sheep as a model, we performed a comparative transcriptome analysis between in vivo (IVO; in vivo fertilized followed by further development in the uterus) and in vitro produced (IVP; IVF with further culture in the incubator) conceptuses, and the caruncular and intercaruncular areas of the ovine endometrium. We identified several genes that were differentially expressed between the IVO and IVP groups on day 17, when adhesion between the trophoblast and the uterine luminal epithelium begins in sheep. By performing Gene Ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, we found that, in the conceptus, differentially expressed genes (DEGs) were associated mainly with functions relating to cell binding and the cell cycle. In the endometrial caruncular area, DEGs were involved in cell adhesion/migration and apoptosis, and in the intercaruncular area, they were significantly enriched in pathways of signal transduction and transport. Thus, these DEGs are potential candidates for further exploring the mechanism underlying IVF/IVP-induced embryonic implant failure that occurs due to a loss of interaction between the conceptus and endometrium during the peri-implantation period.

Variations in phthalate ester (PAE) accumulation and their formation mechanism in Chinese flowering cabbage (Brassica parachinensis L.) cultivars grown on PAE-contaminated soils.

Phthalate ester (PAE) accumulation in crops poses great risks to human health and has aroused great concern. Here, we investigated variations in di-n-butylphthalate (DBP) and di-(2-ethylhexyl) phthalate (DEHP) accumulation by various Chinese flowering cabbage cultivars and revealed their variation mechanism. There were significant differences (p < 0.05) in shoot PAE concentrations of 28 cultivars. Moreover, significant positive correlations between DBP and DEHP concentrations in shoots of all cultivars indicated that they could be taken up simultaneously by various cultivars. Due to the lower translocation factor of low-PAE accumulator, its shoot PAEs concentrations were much lower than root compared to high-PAE accumulator. Further, subcellular distribution showed that PAE concentrations of root cell walls and organelles were much higher than those of shoots in low-PAE accumulator. Therefore, lower translocation from root to shoot and more PAEs accumulating in cell walls and organelles of root might act as main formation mechanism of low-PAE accumulator.

H1foo is essential for in vitro meiotic maturation of bovine oocytes.

Oocyte-specific linker histone, H1foo, is localized on the oocyte chromosomes during the process of meiotic maturation, and is essential for mouse oocyte maturation. Bovine H1foo has been identified, and its expression profile throughout oocyte maturation and early embryo development has been established. However, it has not been confirmed if H1foo is indispensable during bovine oocyte maturation. Effective siRNAs against H1foo were screened in HeLa cells, and then siRNA was microinjected into bovine oocytes to down-regulate H1foo expression. H1foo overexpression was achieved via mRNA injection. Reverse transcription polymerase chain reaction (RT-PCR) results indicated that H1foo was up-regulated by 200% and down-regulated by 70%. Based on the first polar body extrusion (PB1E) rate, H1foo overexpression apparently promoted meiotic progression. The knockdown of H1foo significantly impaired bovine oocyte maturation compared with H1foo overexpression and control groups (H1foo overexpression = 88.7%, H1foo siRNA = 41.2%, control = 71.2%; P < 0.05). This decrease can be rescued by co-injection of a modified H1foo mRNA that has escaped from the siRNA target. However, the H1e (somatic linker histone) overexpression had no effect on PB1E rate when compared with the control group. Therefore we concluded that H1foo is essential for bovine oocyte maturation and its overexpression stimulates the process.

Reconstruction of damaged corneal epithelium using Venus-labeled limbal epithelial stem cells and tracking of surviving donor cells.

Limbal epithelial stem cells are responsible for the self-renewal and replenishment of the corneal epithelium. Although it is possible to repair the ocular surface using limbal stem cell transplantation, the mechanisms behind this therapy are unclear. To investigate the distribution of surviving donor cells in a reconstructed corneal epithelium, we screened a Venus-labeled limbal stem cell strain in goats. Cells were cultivated on denuded human amniotic membrane for 21 days to produce Venus-labeled corneal epithelial sheets. The Venus-labeled corneal epithelial sheets were transplanted to goat models of limbal stem cell deficiency. At 3 months post-surgery, the damaged corneal epithelia were obviously improved in the transplanted group compared with the non-transplanted control, with the donor cells still residing in the reconstructed ocular surface epithelium. Using Venus as a marker, our results indicated that the location and survival of donor cells varied, depending on the corneal epithelial region. Additionally, immunofluorescent staining of the reconstructed corneal epithelium demonstrated that many P63(+) cells were unevenly distributed among basal and suprabasal epithelial layers. Our study provides a new model, and reveals some of the mechanisms involved in corneal epithelial cell regeneration research.

Replacement of H1 linker histone during bovine somatic cell nuclear transfer.

Linker histone variants are involved in regulation of chromosome organization and gene transcription; several subtypes are expressed in the maturing oocyte and developing embryo. In Xenopus and mice, the transition between linker histone variants occurred following nuclear transfer, and apparently contributed to donor nuclear reprogramming. To determine whether such linker histone replacement occurred after bovine nuclear transfer, red fluorescent protein (RFP) tagged H1e (somatic linker histone H1e) donor cells and Venus tagged H1foo eggs were created, enucleated eggs were injected with donor cells, and embryos were created by fusion. Using fluorescence microscopy, release of H1e in the donor nucleus, acquisition of H1foo by donor chromosomes, and the H1foo-to-H1e transition were observed in live cells. Linker histone replacement occurred more slowly in bovine than murine embryos. Low levels of diffuse red fluorescence (H1e) in the donor nucleus were detected 5 h after fusion, at which time green fluorescence (H1foo) had incorporated into donor chromosomes. However, complete replacement did not occur until 8 h after fusion. We concluded that the linker histone transition was sufficiently conserved among species, which provided further evidence regarding its important role in nuclear reprogramming.

Evaluation of quality of Radix Puerariae herbal medicine by isoflavonoids.

OBJECTIVES: A high-performance liquid chromatography (HPLC) method was developed to examine five isoflavonoids present in Chinese herbal medicinal products containing Radix Puerariae. METHODS: Five isoflavonids, puerarin, daidzin, genistin, daidzein and genistein, were measured by HPLC. The HPLC system was equipped with an ODS-AM-303 column (250 mm x 4.6 mm i.d., 5 microm) and established gradient system comprising glacial acetic acid/water and glacial acetic acid/acetonitrile. KEY FINDINGS: The developed HPLC system yielded good separation of the five isoflavonoids. Relative coefficients of intraday and interday analysis of variation were less than 5%. The isoflavonoid recovery from Radix Puerariae was 90-113%. Most of the Radix Puerariae products studied contained five isoflavonoids in their HPLC fingerprint. The major component was purarine, then daidzin and daidzein; genistin and genistein were the least abundant. Five Radix Puerariae herbal medicines contained various concentrations of isoflavonoids. Of the 11 scientific extracted formulas of Radix Puerariae tested, ST brands had a greater isoflavonoid content than KA and SC brands. CONCLUSIONS: Separation and quantification of the five isoflavonoids by this HPLC method was suitable to assess the quality of Radix Puerariae herbal medicine products.