Establishment and optimization of sulfur-based autotrophic-heterotrophic denitrification biofilters for advanced post-anaerobic treatment of effluent from kitchen wastewater and landfill leachate under low temperature.

Landfill leachate treatment is a major challenge in wastewater treatment. In this study, two sulfur-based autotrophic-heterotrophic denitrification biofilters (Ra biofilter with room-temperature molded filler and Rb biofilter with melt molded filler) were used to treat kitchen-landfill leachate at low temperatures. The effects of reflux ratio, concentrations of NaHCO3, and Na2S2O3 on the total nitrogen removal efficiency were analyzed, and based on response surface methodology, the optimum parameters were determined. After optimization, the total nitrogen removal efficiency for the Ra and Rb biofilters increased by 83% and 81%, respectively. Moreover, sulfur-based autotrophic denitrification accounted for more than 70% of the nitrogen removal in both biofilters. Based on high-throughput sequencing results, the functional bacteria exhibited high abundance in the Ra biofilter, indicating that the room-temperature molded filler favored the enrichment of functional bacteria. These findings were important for optimizing the operation of sulfur autotrophic-heterotrophic denitrification biofilters at low temperatures.

Familiarity of teaching skills among general practitioners transfer training trainers in China: a cross-sectional survey.

BACKGROUND: The insufficient number of general practitioners (GPs) is a major challenge facing China's healthcare system. The purpose of the GP transfer training programme was to provide training for experienced doctors to transition to general practice. However, research on the competencies of GP transfer training trainers in teaching skills in China is limited. This cross-sectional study aimed to examine the baseline familiarity with teaching skills among Chinese GP transfer training trainers. METHODS: An online survey was conducted among trainers who participated in the 2021 Sichuan Province General Practice Training Trainer Program. The survey collected data on participants' characteristics and familiarity with 20 skills in three essential teaching knowledge areas: the core functions of primary care (five questions), preparation for lesson plan (four questions), and teaching methods (11 questions). RESULTS: In total, 305 participants completed the survey. Familiarity rates were generally low across all three essential teaching knowledge areas. No significant differences were observed in familiarity rates between the tertiary and secondary hospitals. CONCLUSION: This study revealed gaps in the teaching skills of GP transfer training trainers in China. These results suggest the necessity for targeted training programs to enhance the teaching skills and competencies of trainers.

Cr(VI) removal from contaminated waters using ultra-thin layered meixnerite.

Cr(VI) is of great concern to public health and environmental safety due to its high toxicity. Here, we report a low-cost yet highly efficient method to prepare a novel LDH, ultra-thin layered meixnerite, which performed superiorly in treatment of aqueous Cr(VI) with little secondary pollution being induced. The produced ultra-thin layered meixnerite was composed of nanoparticles with a thickness of around 7 nm, less than 9 times the thickness of a single LDH layer. The XRD patterns of the ultra-thin layered meixnerite, in which the characteristic diffraction peaks of a typical LDH were weakened or even disappeared, confirmed the successful delamination. This special morphology of the ultra-thin layered meixnerite was not only helpful to its full dispersion in the Cr(VI)-bearing solutions but also facilitated the formation of more active sorption sites on its external surface. As a result, the maximum sorption capacity of UTLM for Cr(VI) removal was 480.9 mg g-1, far higher than that of OM (196.9 mg g-1). In addition to electrostatic attraction and anion exchange, the ultra-thin layered meixnerite could also become restacked during removal of aqueous Cr(VI) to generate inner-sphere complexation, finally inducing an enhanced Cr(VI) uptake. Furthermore, XPS analysis characterized the promotion of the break of Al-OH bond with the increase in temperature, and the Cr-O peak increased correspondingly from 29.69% at 25 °C to 48.77% at 85 °C, resulting that the ultra-thin layered meixnerite could remove Cr(VI) more effectively at higher reaction temperatures. Therefore, ultra-thin layered meixnerite is very suitable for future application in treatment of industrial wastewaters with elevated temperatures.

Computation of Binding Energy of MCS and GO-Grafted MCS with Waterborne Epoxy Resin Using Density Functional Theory Method: Investigating the Corrosion Resistance of the Composite Coatings.

In order to overcome the problems of poor corrosion resistance and low hydrophobicity of water-based coatings. Two corrosion-inhibiting materials, graphene oxide (GO) and modified chitosan (MCS), were added to the coatings to obtain a new type of coating with comprehensive properties. The composite material formed by PVA cross-linked waterborne epoxy resin was named "substrate". The density functional theory (DFT) calculation was used to explore the binding ability of MCS and GO-grafted MCS to the substrate, respectively. The results showed that the complex cross-linked network structure formed by the grafting of GO and MCS not only improved the intermolecular interaction force but also improved the binding ability to the substrate, and the coating is denser, effectively delaying the erosion to the coating by the corrosive medium. The composite coating exhibited excellent dual functional properties of hydrophobicity and corrosion resistance at the coating-metal interface, and a stronger protective effect was formed upon the steel plate. Studies showed that this composite coating has good hydrophobic properties. (The contact angle of the composite waterborne coating reaches 87°.) It also has low self-corrosion current (0.28/cm-2) and high corrosion voltage (-0.45 V). The maximum inhibition efficiency of the coating is 99.97%.

DCBLD2 regulates vascular hyperplasia by modulating the platelet derived growth factor receptor-ß endocytosis through Caveolin-1 in vascular smooth muscle cells.

DCBLD2 is a neuropilin-like transmembrane protein that is up-regulated during arterial remodeling in humans, rats, and mice. Activation of PDGFR-ß via PDGF triggers receptor phosphorylation and endocytosis. Subsequent activation of downstream signals leads to the stimulation of phenotypic conversion of VSMCs and arterial wall proliferation, which are common pathological changes in vascular remodeling diseases such as atherosclerosis, hypertension, and restenosis after angioplasty. In this study, we hypothesized that DCBLD2 regulates neointimal hyperplasia through the regulation of PDGFR-ß endocytosis of vascular smooth muscle cells (VSMCs) through Caveolin-1 (Cav-1). Compared with wild-type (WT) mice or control littermate mice, the germline or VSMC conditional deletion of the Dcbld2 gene resulted in a significant increase in the thickness of the tunica media in the carotid artery ligation. To elucidate the underlying molecular mechanisms, VSMCs were isolated from the aorta of WT or Dcbld2-/- mice and were stimulated with PDGF. Western blotting assays demonstrated that Dcbld2 deletion increased the PDGF signaling pathway. Biotin labeling test and membrane-cytosol separation test showed that after DCBLD2 was knocked down or knocked out, the level of PDGFR-ß on the cell membrane was significantly reduced, while the amount of PDGFR-ß in the cytoplasm increased. Co-immunoprecipitation experiments showed that after DCBLD2 gene knock-out, the binding of PDGFR-ß and Cav-1 in the cytoplasm significantly increased. Double immunofluorescence staining showed that PDGFR-ß accumulated Cav-1/lysosomes earlier than for control cells, which indicated that DCBLD2 gene knock-down or deletion accelerated the endocytosis of PDGF-induced PDGFR-ß in VSMCs. In order to confirm that DCBLD2 affects the relationship between Cav-1 and PDGFR-ß, proteins extracted from VSMCs cultured in vitro were derived from WT and Dcbld2-/- mice, whereas co-immunoprecipitation suggested that the combination of DCBLD2 and Cav-1 reduced the bond between Cav-1 and PDGFR-ß, and DCBLD2 knock-out was able to enhance the interaction between Cav-1 and PDGFR-ß. Therefore, the current results suggest that DCBLD2 may inhibit the caveolae-dependent endocytosis of PDGFR-ß by anchoring the receptor on the cell membrane. Based on its ability to regulate the activity of PDGFR-ß, DCBLD2 may be a novel therapeutic target for the treatment of cardiovascular diseases.

FT-like paralogs are repressed by an SVP protein during the floral transition in Phalaenopsis orchid.

KEY MESSAGE: An SVP protein, PhSVP, bound to the CArG-boxes in the promoter regions of FT-like paralogs and repressed their expression, thus affecting the floral transition in Phalaenopsis orchid. Phalaenopsis is an important ornamental flower native to tropical rain forests. It usually reaches vegetative maturity after 4-5 leaves and, after a juvenile stage, forms a flower spike (inflorescence) from the axillary buds. The PEBP gene family encodes a phosphatidyl-ethanolamine-binding protein (PEBP) domain involved in regulating flowering and other aspects of plant development. Here, we identified eight PEBP family genes in Phalaenopsis and detected the expression patterns of seven of them in various organs. Among them, PhFT1 (Phalaenopsis hybrid FLOWERING LOCUS T1), PhFT3, PhFT5, and PhMFT (Phalaenopsis hybrid MOTHER OF FT AND TFL1) promoted flowering in transgenic Arabidopsis, while PhFT6 inhibited flowering. PhSVP (Phalaenopsis hybrid SHORT VEGETATIVE PHASE), an SVP protein that repressed flowering in Arabidopsis, bound to the CArG-boxes in the promoter regions of PhFT3, PhFT6, and PhMFT in a yeast one-hybrid assay. Additionally, dual-luciferase and transient expression assays showed that PhSVP significantly inhibits the expression of both PhFT3 and PhFT6. Together, our work provides a comprehensive understanding of the PhFT-like genes that can promote or repress flowering, and it suggests strategies for regulating the floral transition in Phalaenopsis that exploit the evolutionary versatility of PhFTs to respond to various signals stimuli.

Association of serum CTRP9 levels with cardiac autonomic neuropathy in patients with type 2 diabetes mellitus.

AIMS: Cardiac autonomic neuropathy (CAN) is a serious complication of diabetes and is associated with adipokines. The C1q tumor necrosis factor-related protein 9 (CTRP9) is a newly discovered adipokine. This study aimed to evaluate the association of serum CTRP9 levels with the prevalence and severity of CAN in patients with type 2 diabetes mellitus. MATERIALS AND METHODS: We enrolled 262 patients (aged ≥18 years) with type 2 diabetes mellitus into this study. Standard cardiovascular autonomic reflex tests (CARTs) were used to assess CAN and patients were divided into three groups accordingly: a non-CAN group, an early CAN group, and a definite CAN group. Serum CTRP9 levels were measured by enzyme-linked immunosorbent assay, and the tertiles were calculated. RESULTS: Serum CTRP9 levels decreased significantly in the early CAN and definite CAN groups (P < 0.05). The percentage of definite CAN was the highest at the minimum tertile of serum CTRP9 level (T1; P < 0.05). Additionally, serum CTRP9 levels were negatively correlated with age, DM duration, hemoglobin A1c (HbA1c), and fasting plasma glucose (FPG) while positively correlated with high-density lipoprotein cholesterol (HDL; P < 0.05). The level of CTRP9 was also significantly associated with the four indexes of CARTs (P < 0.05). Furthermore, CTRP9 was a protective factor for definite CAN (P < 0.05). Compared with the maximum tertile (T3) of the serum CTRP9 levels, a decreased level of serum CTRP9 in T1 significantly increased the prevalence ratio of definite CAN in patients with type 2 diabetes mellitus (P < 0.05). CONCLUSION: Serum CTRP9 levels were independently associated with definite CAN. CTRP9 represents a reliable biomarker for exploring CAN in patients with type 2 diabetes mellitus.

The Effect of L-Carnitine Additive During In Vitro Maturation on the Vitrification of Pig Oocytes.

Cryopreservation of oocytes/embryos is an important technique for genetic resources; however, the success of vitrification in pig oocytes remained at a relatively lower level due to the high content of lipid droplets (LDs). Considering the positive effect of L-carnitine on the function of LDs, the present study was designed to investigate the effect of the addition of L-carnitine on the vitrification of porcine cumulus cells of complexes (cumulus/oocyte complexes [COCs]). First, COCs were randomly divided into two groups: one group of COCs were commonly in vitro maturation (IVM) for 42-46 hours (nonvitrification [NV]), while another group of COCs were IVM with 10 mM L-carnitine (NVL [nonvitrification with L-carnitine addition in IVM]). In addition, random parts of COCs with L-carnitine addition were vitrified (VL [vitrification with L-carnitine addition in IVM]), while vitrification was performed on COCs without L-carnitine used as control group (V). Results showed that the maturation rate of pig oocytes reduced significantly when the vitrification was performed at 16 hours during IVM (VL vs. NVL, 40.09 ± 2.85 vs. 90.76 ± 1.16; V vs. NV, 34.41 ± 2.55 vs. 89.71 ± 1.33, p < 0.01). With the addition of L-carnitine, intracellular LDs were decreased significantly (p < 0.01). However, no difference was observed on the efficiency of vitrification in pig oocytes (VL vs. V, 40.09 ± 2.85 vs. 34.41 ± 2.55, p > 0.05). In addition, not only the reactive oxygen species (ROS) level in pig oocytes with the L-carnitine addition group reduced significantly (p < 0.01), but also the expression of SOD1 gene was improved (p < 0.05). In conclusion, results demonstrated that although no difference could be observed on pig COC vitrification, the LDs and ROS level in pig oocytes could be modified by the addition of L-carnitine, which might be helpful for further development.

Comparative Study of Growth, Cadmium Accumulation and Tolerance of Three Chickpea (Cicer arietinum L.) Cultivars.

Trace metals (TM) contamination is a severe problem in the environment and produced an adverse effect on the productivity of crops. Cadmium (Cd) is a TM ranked seven among the top 20 pollutants due to its high toxicity and solubility in water, taken up by the plants and affects their growth and metabolism. In this study, we evaluated the growth, Cd accumulation and tolerance capacities of three chickpea (Cicer arietinum L.) cultivars (NC234 (NC2), ICCV89310 (IC8) and ICCV89323-B (IC8-B)), subjected to two Cd concentrations (25 and 50 µM) in hydroponic culture. The toxicity of Cd reduced the plant height and fresh and dry biomass in all cultivars. The maximum reduction was observed at 50 µM of Cd. Compared with IC8-B, cultivars IC8 and NC2 exhibited better performance with high growth, biomass, root to shoot (R/S) ratio and water content under high Cd stress. To measure the accumulation of Cd in root and shoot, an inductively coupled plasma optical emission spectrometer (ICP-OES) was used. IC8 and NC2 had comparatively high Cd tolerance and accumulation ability (> 100 µg g-1 dry weight), with IC8 being more tolerant and accumulated higher Cd in shoot than NC2, while cultivar IC8-B was sensitive. Root accumulated more Cd than shoot in a dose-dependent manner. The bioconcentration factors (BCF) and bioaccumulation coefficients (BAC) were far higher than one (> 1) and increased with an increase in Cd concentrations, while the translocation factor (TF) was less than one (< 1), suggesting that all the three cultivars were unable to transfer Cd from the root to the shoot efficiently. Our results indicated that IC8 and NC2 proved to be resistant, while IC8-B showed sensitivity when exposed to high Cd stress (50 µM).

Integrated analysis of transcriptome data revealed MMP3 and MMP13 as critical genes in anaplastic thyroid cancer progression.

To better understand the molecular mechanisms of anaplastic thyroid carcinoma (ATC), we aimed to identify the hub genes specifically involved in ATC by integrated bioinformatics analysis. In this study, using three Gene Expression Omnibus data sets with the same platform GPL570, we screened hub genes involved in ATC progression. In vitro experiments, such as western blot analysis, Transwell assays, and coimmunoprecipitation, was performed to verify our findings. By comparing three subtypes of thyroid cancer with normal tissue, we found ATC harbored more changed genes than well and poorly differentiated thyroid cancer. Using specifically differentially expressed genes between ATC and normal thyroid tissues to perform Gene ontology (GO) analysis, ATC showed enrichments of GO terms involved in lymphocyte migration and activation, collagen catabolic and metabolic process, thyroid hormone synthesis, and embolism. Using genes involved in extracellular matrix, coexpression network analysis and protein-protein interaction analysis were performed to identify matrix metalloproteinase 3 (MMP3) and MMP13 as two hub genes. Our experimental data indicated that both MMP3 and MMP13 were upregulated in ATC and knockdown of either of them could notably suppress ATC cell invasion and migration. Mechanistically, Gene Set Enrichment Analysis, coimmunoprecipitation, and rescue experiments revealed MMP3 and MMP13 not only interacted with each other, but also regulated each other through the janus kinase/signal transducer and activator of transcription 3 and mammalian target of rapamycin pathways. In conclusion, we identified a specific molecular mechanisms for the development of ATC by integrated analysis of transcriptome and in vitro experiments, which suggested that MMP3 and MMP13 might be developed as novel therapeutic targets for ATC.

Research of activity of Main Belt Comets 176P/LINEAR, 238P/Read and 288P/(300163) 2006 VW139.

As a new class of comet, main belt comets (MBCs) have attracted more and more attention in recent years. To study activity and physical properties of three MBCs 176P/LINEAR, 238P/Read and 288P/(300163) 2006 VW139, we carried out broadband CCD photometry of three MBCs on UT 2016 November 18-19 with the 1-m optical telescope at Lulin Observatory in Taiwan. By comparing cometary surface brightness profiles to stellar surface brightness profiles, and by comparing cometary absolute magnitude to the expected magnitude of inactive nucleus, we found that 176P/LINEAR was inactive, while 238P/Read and 288P/(300163) 2006 VW139 were active. By photometric studies, we obtained the Afρ values and the dust production rates. Finally, the activity of three MBCs were discussed. Our photometric results show that the total dust mass of 238P/Read and 288P/(300163) 2006 VW139 obtained in this work are of the same magnitude as the majority of known MBCs.

KLF5 promotes the tumorigenesis and metastatic potential of thyroid cancer cells through the NF-κB signaling pathway.

The purpose of the present study was to identify the potential function of Kruppel-like factor 5 (KLF5) in thyroid cancer and investigate the underlying mechanisms. The protein levels of KLF5 in 98 thyroid cancer tissues were analyzed using an immunohistochemistry assay. SW579 cells transfected with small interfering RNA against KLF5 and B-CPAP cells transfected with KLF5 expressing vectors were used for functional studies. Western blot analysis, immunofluorescence and co-immunoprecipitation assays were used to investigate the mechanisms of KLF5. In vivo tumorigenicity was assessed using a subcutaneous xenograft experiment. The results revealed that KLF5 was highly expressed in thyroid cancer tissues and associated with lymph node metastasis. Knockdown of KLF5 in SW579 cells suppressed proliferation, anchorage-independent growth, migration and invasion in vitro, while the overexpression of KLF5 resulted in opposite effects in B-CPAP cells. Mechanistically, it was demonstrated that KLF5 promoted the cytoplasm-nuclear translocation of nuclear factor-κB. Additionally, it was revealed that insufficient F-box/WD repeat-containing protein 7 expression may be responsible for the dysfunction of KLF5 in thyroid cancer. These results revealed that KLF5 promotes the tumorigenesis and metastasis of thyroid cancer cells and may be a potential therapeutic target in patients with thyroid cancer.

Genome-Wide Association and Functional Studies Identify SCML4 and THSD7A as Novel Susceptibility Genes for Coronary Artery Disease.

OBJECTIVE: The genetic contribution to coronary artery disease (CAD) remains largely unclear. We combined genetic screening with functional characterizations to identify novel loci and candidate genes for CAD. APPROACH AND RESULTS: We performed genome-wide screening followed by multicenter validation in 8 cohorts consisting of 21 828 participants of Han ethnicity and identified 3 novel intragenic SNPs (single nucleotide polymorphisms), rs9486729 (SCML4 [Scm polycomb group protein-like 4]; odds ratio, 1.25; 95% CI, 1.17-1.34; P=3.51×10-11), rs17165136 (THSD7A [thrombospondin type 1 domain-containing 7A]; odds ratio 1.28; 95% CI, 1.21-1.35; P<1.00×10-25), and rs852787 (DAB1 [disabled-1]; odds ratio, 1.29; 95% CI, 1.21-1.38; P=2.02×10-14), associated with CAD with genome-wide significance. The risk allele of rs9486729 and protective allele of rs17165136 were associated with the decreased expression of their host genes, SCML4 and THSD7A, respectively, whereas rs852787 did not have transcriptional effects on any gene. Knockdown of SCML4 activated endothelial cells by increasing the expression of IL-6, E-selectin, and ICAM and weakened their antiapoptotic activity, whereas the knockdown of THSD7A had little effect on these endothelial cell functions but attenuated monocyte adhesion via decreasing the expression of ICAM, L-selectin, and ITGB2. We further showed that inhibiting the expression of SCML4 exacerbated endothelial dysfunction and vascular remodeling in a rat model with partial carotid ligation. CONCLUSIONS: We identify 3 novel loci associated with CAD and show that 2 genes, SCML4 and THSD7A, make functional contributions to atherosclerosis. How rs852787 and its host gene DAB1 are linked to CAD needs further studies.

Mechanism and early intervention research on ALI during emergence surgery of Stanford type-A AAD: Study protocol for a prospective, double-blind, clinical trial.

BACKGROUND: Stanford type-A acute aortic dissection (AAD) is a severe cardiovascular disease demonstrating the characteristics of acute onset and rapid development, with high morbidity and mortality. The available evidence shows that preoperative acute lung injury (ALI) induced by Stanford type-A AAD is a frequent and important cause for a number of untoward consequences. However, there is no study assessing the incidence of preoperative ALI and its independent determinants before Standford type-A AAD surgery in Chinese adult patients. METHODS/DESIGN: This is a prospective, double-blind, signal-center clinical trial. We will recruit 130 adult patients undergoing Stanford type-A AAD surgery. The incidence of preoperative ALI will be evaluated. Perioperative clinical baselines and serum variables including coagulation, fibrinolysis, inflammatory, reactive oxygen species, and endothelial cell function will be assayed. The independent factors affecting the occurrence of preoperative ALI will be identified by multiple logistic regression analysis. TRIAL REGISTRATION: ClinicalTrials.gov (https://register.clinicaltrials.gov/), Registration number NCT01894334.

Hypoacetylation, hypomethylation, and dephosphorylation of H2B histones and excessive histone deacetylase activity in DU-145 prostate cancer cells.

BACKGROUND: Hypoacetylation on histone H3 of human prostate cancer cells has been described. Little is known about the modifications of other histones from prostate cancer cells. METHODS: Histones were isolated from the prostate cancer cell line DU-145 and the non-malignant prostatic cell line RC170N/h. Post-translational modifications of histone H2B were determined by liquid chromatography-mass spectrometry (LC-MS)/MS. RESULTS: The histone H2B of the prostate cancer cell line DU-145 was found to have hypoacetylation, hypomethylation, and dephosphorylation as compared to the non-malignant prostatic cell line RC170N/h. H2B regained acetylation on multiple lysine residues, phosphorylation on Thr19, and methylation on Lys23 and Lys43 in the DU-145 cells after sodium butyrate treatment. CONCLUSIONS: The histone H2B of DU-145 prostate cancer cells are hypoacetylated, hypomethylated, and dephosphorylated. Histone deacetylase inhibitor reversed this phenotype. Epigenetic agent may therefore be useful for prostate cancer therapy and worth further investigation.

Osteoinductive factor is a novel biomarker for the diagnosis of early diabetic nephropathy.

BACKGROUND: Microalbuminuria is the earliest clinical sign of diabetic nephropathy (DN). However, earlier markers as a diagnostic tool for DN was required for the invalid of microalbuminuria in some cases. Osteoinductive factor (OIF) was known to be an essential component of the normal vascular matrix. We aimed to research the relationship between DN and OIF, and discussed the availability of the serological markers for earlier stage of DN. METHOD: One hundred twenty Chinese subjects, who included patients with type 2 diabetes mellitus (T2DM), DN with microalbuminuria, and DN with macroalbuminuria, as well as healthy controls, were enrolled in this study. Serum OIF levels were examined by ELISA and other clinical biochemical parameters were tested based on standard methods. RESULTS: Our results indicated that, serum OIF levels were significantly increased in DN subjects compared with healthy and T2DM subjects (P<0.05 respectively). However, no significant changes in serum OIF levels were found between T2DM and healthy subjects. Furthermore, serum OIF had negative correlation with estimated glomerular filtration rate (eGFR) and positive correlation with blood urea nitrogen(BUN) and creatinine. ROC curve analysis showed that serum OIF level was a good sensitive and specificity marker for microalbuminuria and early renal damage with sensitivity of 86.7% and specificity of 95%, as well as for macroalbuminuria and damage progress with sensitivity of 90% and specificity of 95%. CONCLUSION: OIF may be an indicator of the earlier-stage DN in subjects with T2DM. Understanding the exact mechanism of up-regulated OIF in subjects with DN requires further study.

Phenotype and transcriptome analysis reveals chloroplast development and pigment biosynthesis together influenced the leaf color formation in mutants of Anthurium andraeanum 'Sonate'.

Leaf color is one of the well-sought traits in breeding program for Anthurium andraeanum Lind. Knowledge of mechanisms in anthuriums to produce leaves with different shades of green would help to effectively select desirable traits. In this study, the micro- and ultra-structural and physiological features of leaves on wild type and leaf color mutants (dark green, rubescent, etiolated, albino) in A. andraeanum 'Sonate' were analyzed. Results show that chloroplasts of leaf color mutants exhibited abnormal morphology and distribution. Using next generation sequencing technology followed by de novo assembly, leaf transcriptomes comprising of 41,017 unigenes with an average sequence length of 768 bp were produced from wild type and rubescent mutant. From the 27,539 (67.1%) unigenes with annotated functions, 858 significantly differently expressed genes (DEGs) were identified, consisting of 446 up-regulated genes and 412 down-regulated genes. Genes that affect chloroplasts development and division, and chlorophyll biosynthesis were included in the down-regulated DEGs. Quantitative real-time PCR (qRT-PCR) analysis validated that the expression level of those genes was significantly lower in the rubescent, etiolated, and albino mutant compared to wild type plants, which concurs with the differences in micro- and ultra-structures and physiological features between these two types of plants. Conclusively, the leaf color formation is greatly affected by the activity of chloroplast development and pigment biosynthesis. And the possible formation pathway of leaf color mutant of A. andraeanum 'Sonate' is deduced based on our results.

[Analysis of CYP21A2 gene mutations in two families with 21-hydroxylase deficiency].

OBJECTIVE: To analyze CYP21A2 gene mutation in two families with 21-hydroxylase deficiency (21-OHD) and to explore the correlation between genotype and clinical phenotype. METHODS: Two patients with 21-OHD and their families were investigated. CYP21A2 gene mutation was analyzed by PCR and direct sequencing. RESULTS: The probands from family 1 and 2 have been respectively diagnosed with simple virilizing and non-classical 21-OHD. Both showed increased baseline serum 17hydroxyprogesterone, testosterone and adrenocorticotropic hormone (ACTH), but had no evidence of salt loss. Computer tomography revealed bilateral adrenal hyperplasia in both patients. After 1 year treatment, both had conceived successfully. DNA sequencing revealed that the proband of family 1 had compound heterozygous mutations for IVS2 13 A>G and Ile172Asn. Her father was heterozygous for Ile172Asn, whilst her mother and brother were heterozygous for IVS213A/C>G. In family 2, the proband was heterozygous for Arg341Trp and Gln318X. Her father, sister and nephew were heterozygous for Arg341Trp, whilst her mother was heterozygous for Gln318X. her brother and niece were non-affected. Carriers of single heterozygous mutations in both families had no clinical sign. CONCLUSION: In both families, the disease has been caused by compound heterozygous mutations, for which there has been a good genotype-phenotype agreement. Screening of CYP21A2 gene can facilitate both diagnosis and genetic counseling.

Phenethyl isothiocyanate and paclitaxel synergistically enhanced apoptosis and alpha-tubulin hyperacetylation in breast cancer cells.

Combination of phenethyl isothiocyanate (PEITC) and paclitaxel (taxol) has been shown to work synergistically to increase apoptosis and cell cycle arrest in breast cancer cells. In this report, we further explored the mechanisms for the synergistic activity of PEITC and taxol in MCF7 and MDA-MB-231 (MB) breast cancer cell lines. By Western blotting analysis, treatment of MCF7 cells with both PEITC and taxol led to a 10.4-fold and 5.96-fold increase in specific acetylation of alpha-tubulin over single agent PEITC and taxol, respectively. This synergistic effect on acetylation of alpha-tubulin was also seen in MB cells. The combination of PEITC and taxol also reduced expressions of cell cycle regulator Cdk1, and anti-apoptotic protein bcl-2, enhanced expression of Bax and cleavage of PARP proteins. In conclusion, this study provided biochemical evidence for the mechanism of synergistic effect between the epigenetic agent PEITC and the chemotherapeutic agent taxol.

PPAR-γ agonist pioglitazone prevents apoptosis of endothelial progenitor cells from rat bone marrow.

Selective peroxisome proliferator-activated receptor-γ (PPAR-γ) agonist affects the functions of endothelial progenitor cells (EPCs). This study explores the effect of selective PPAR-γ agonist, pioglitazone, on EPC apoptosis. The cells were cultured and identified via the double staining method in a medium containing different concentrations of pioglitazone. EPC apoptosis was detected by flow cytometry. On Day 7, EPCs engulfed DiL-ac-LDL and FITC-UEA-1, and showed yellow fluorescence in a laser-scanning confocal microscope. EPC apoptosis inhibition was maximal at 50 µmol/L. The ability of pioglitazone to prevent EPC apoptosis may be mediated by the PI3K/Akt signal pathway. The use of thiazolidine two ketone (TZD) to reduce EPC apoptosis may have some potential in treating vascular diseases.