Genome-wide identification and expression profiling of the carotenoid cleavage dioxygenase (CCD) gene family in Brassica napus L.

Carotenoid cleavage dioxygenase (CCD), a key enzyme in carotenoid metabolism, cleaves carotenoids to form apo-carotenoids, which play a major role in plant growth and stress responses. CCD genes had not previously been systematically characterized in Brassica napus (rapeseed), an important oil crop worldwide. In this study, we identified 30 BnCCD genes and classified them into nine subgroups based on a phylogenetic analysis. We identified the chromosomal locations, gene structures, and cis-promoter elements of each of these genes and performed a selection pressure analysis to identify residues under selection. Furthermore, we determined the subcellular localization, physicochemical properties, and conserved protein motifs of the encoded proteins. All the CCD proteins contained a retinal pigment epithelial membrane protein (RPE65) domain. qRT-PCR analysis of expression of 20 representative BnCCD genes in 16 tissues of the B. napus cultivar Zhong Shuang 11 ('ZS11') revealed that members of the BnCCD gene family possess a broad range of expression patterns. This work lays the foundation for functional studies of the BnCCD gene family.

Differential Alternative Splicing Genes and Isoform Regulation Networks of Rapeseed (Brassica napus L.) Infected with Sclerotinia sclerotiorum.

Alternative splicing (AS) is a post-transcriptional level of gene expression regulation that increases transcriptome and proteome diversity. How the AS landscape of rapeseed (Brassica napus L.) changes in response to the fungal pathogen Sclerotinia sclerotiorum is unknown. Here, we analyzed 18 RNA-seq libraries of mock-inoculated and S. sclerotiorum-inoculated susceptible and tolerant B. napus plants. We found that infection increased AS, with intron retention being the main AS event. To determine the key genes functioning in the AS response, we performed a differential AS (DAS) analysis. We identified 79 DAS genes, including those encoding splicing factors, defense response proteins, crucial transcription factors and enzymes. We generated coexpression networks based on the splicing isoforms, rather than the genes, to explore the genes' diverse functions. Using this weighted gene coexpression network analysis alongside a gene ontology enrichment analysis, we identified 11 modules putatively involved in the pathogen defense response. Within these regulatory modules, six DAS genes (ascorbate peroxidase 1, ser/arg-rich protein 34a, unknown function 1138, nitrilase 2, v-atpase f, and amino acid transporter 1) were considered to encode key isoforms involved in the defense response. This study provides insight into the post-transcriptional response of B. napus to S. sclerotiorum infection.

Bilateral lung transplantation after caesarean section in pregnancy with severe pulmonary arterial hypertension: A case report.

RATIONALE: Pulmonary arterial hypertension (PAH) can lead to an increase in right ventricular load and subsequently heart failure, making severe PAH a contraindication for pregnancy. In addition, PAH may worsen during pregnancy and puerperium, which requires high-quality critical care. This report is the first instance in which a patient with severe PAH, survived a successful atrial septal defect (ASD) repair and bilateral lung transplantation during puerperium. PATIENT CONCERNS: A 42-year-old pregnant woman with congenital heart disease (CHD) and severe PAH was admitted to our hospital for the management of pregnancy and delivery. The patient was diagnosed with severe PAH in 2013, and no significant improvements or deteriorations were found until this pregnancy-related hospital admission. DIAGNOSIS: The patient was diagnosed with CHD and severe PAH in 2013 with color Doppler echocardiography, right cardiac catheterization, and pulmonary perfusion imaging. The patient's mean pulmonary arterial pressure increased to 140 mm Hg during pregnancy, suggesting an exacerbated PAH with high risks to both her and the unborn child. INTERVENTIONS: The patient was treated with PAH-targeting treprostinil injection to reduce pulmonary artery pressure. Caesarean section was performed at 27 weeks and 5 days of gestation. The patient was put under extracorporeal membrane oxygenation (ECMO) with the help of local anesthesia before the operation. The investigators finally conducted a bilateral lung transplantation with a shell incision of the sternum under cardiopulmonary bypass. OUTCOMES: The mother and the neonate survived and recovered well after the operation, and were discharged from the hospital on the fourth month post-hospitalization. LESSONS: Severe PAH is an absolute contraindication for pregnancy. However, for patients who insist on a pregnancy, it could be plausible to proceed with a targeted drug therapy and ECMO after conducting a cesarean section, and finally, a lung transplantation. Multidisciplinary diagnosis and treatment is the key to the successful treatment of a PAH-complicated pregnancy.

Brassica napus Infected with Leptosphaeria maculans.

Alternative splicing (AS) is a post-transcriptional regulatory process that enhances transcriptome diversity, thereby affecting plant growth, development, and stress responses. To identify the new transcripts and changes in the isoform-level AS landscape of rapeseed (Brassica napus) infected with the fungal pathogen Leptosphaeria maculans, we compared eight RNA-seq libraries prepared from mock-inoculated and inoculated B. napus cotyledons and stems. The AS events that occurred in stems were almost the same as those in cotyledons, with intron retention representing the most common AS pattern. We identified 1892 differentially spliced genes between inoculated and uninoculated plants. We performed a weighted gene co-expression network analysis (WGCNA) to identify eight co-expression modules and their Hub genes, which are the genes most connected with other genes within each module. There are nine Hub genes, encoding nine transcription factors, which represent key regulators of each module, including members of the NAC, WRKY, TRAF,AP2/ERF-ERF, C2H2,C2C2-GATA, HMG, bHLH, and C2C2-CO-like families. Finally, 52 and 117 alternatively spliced genes in cotyledons and stems were also differentially expressed between mock-infected and infected materials, such as HMG and C2C2-Dof; which have dual regulatory mechanisms in response to L. maculans. The splicing of the candidate genes identified in this study could be exploited to improve resistance to L. maculans.

A Fatal Case of Acute Arsenic Poisoning.

This manuscript reported a case of fatal arsenic poisoning. A woman with schizophrenia took arsenic-containing "pills," which consisted of arsenic trioxide and realgar (arsenic (II) sulfide) and wrapped with gauze. The victim consumed 1.09 and 0.819 g arsenic on two occasions, respectively, with the interval between the two doses of 3 days. The woman died on the sixth day after the first dose without any treatment. In this case, pathological examination revealed fat degeneration of the liver rather than hepatomegaly, a rare finding in acute arsenic poisoning. Arsenic in tissue samples was measured, the total arsenic and inorganic arsenic in blood, liver, and gastric wall was 10.2 µg/mL (9.61 µg/mL), 23.1 µg/g (20.7 µg/g), and 32.3 µg/g (28.6 µg/g), respectively.

Genome-Wide Identification and Expression Profiling of Cytokinin Oxidase/Dehydrogenase (CKX) Genes Reveal Likely Roles in Pod Development and Stress Responses in Oilseed Rape (Brassica napus L.).

Cytokinin oxidase/dehydrogenases (CKXs) play a critical role in the irreversible degradation of cytokinins, thereby regulating plant growth and development. Brassica napus is one of the most widely cultivated oilseed crops worldwide. With the completion of whole-genome sequencing of B. napus, genome-wide identification and expression analysis of the BnCKX gene family has become technically feasible. In this study, we identified 23 BnCKX genes and analyzed their phylogenetic relationships, gene structures, conserved motifs, protein subcellular localizations, and other properties. We also analyzed the expression of the 23 BnCKX genes in the B. napus cultivar Zhong Shuang 11 ('ZS11') by quantitative reverse-transcription polymerase chain reaction (qRT-PCR), revealing their diverse expression patterns. We selected four BnCKX genes based on the results of RNA-sequencing and qRT-PCR and compared their expression in cultivated varieties with extremely long versus short siliques. The expression levels of BnCKX5-1, 5-2, 6-1, and 7-1 significantly differed between the two lines and changed during pod development, suggesting they might play roles in determining silique length and in pod development. Finally, we investigated the effects of treatment with the synthetic cytokinin 6-benzylaminopurine (6-BA) and the auxin indole-3-acetic acid (IAA) on the expression of the four selected BnCKX genes. Our results suggest that regulating BnCKX expression is a promising way to enhance the harvest index and stress resistance in plants.

Meropenem-induced immune thrombocytopenia and the diagnostic process of laboratory testing.

BACKGROUND: Drug-induced immune thrombocytopenia (DITP) is a serious, life-threatening clinical syndrome, the diagnosis of which is consistently difficult. In this report, we present a case of DITP caused by meropenem that was confirmed by laboratory tests. CASE REPORT: A 59-year-old male patient developed severe thrombocytopenia 8 days after the administration of meropenem and cefoperazone-sulbactam. After other causes were ruled out, DITP was suspected. Drug-induced platelet (PLT) antibodies were detected by enzyme immunoassay, flow cytometry, and monoclonal antibody immobilization of PLT antigens (MAIPA). All these tests were performed in the presence and absence of the associated drugs. RESULTS: PLT antibodies were detected in the patient's serum only in the presence of meropenem. MAIPA experiments demonstrated that glycoprotein IIb/IIIa was the binding site of the meropenem-induced PLT antibodies. CONCLUSIONS: Drug-induced immune thrombocytopenia should be considered in cases of acute thrombocytopenia in patients undergoing meropenem treatment. Clinicians should be cognizant of DITP, and a definitive diagnosis should be pursued, if feasible.

Genome-wide analysis and expression profiling of the GRF gene family in oilseed rape (Brassica napus L.).

Growth regulating-factors (GRFs) are plant-specific transcription factors that help regulate plant growth and development. Genome-wide identification and evolutionary analyses of GRF gene families have been performed in Arabidopsis thaliana, Zea mays, Oryza sativa, and Brassica rapa, but a comprehensive analysis of the GRF gene family in oilseed rape (Brassica napus) has not yet been reported. In the current study, we identified 35 members of the BnGRF family in B. napus. We analyzed the chromosomal distribution, phylogenetic relationships (Bayesian Inference and Neighbor Joining method), gene structures, and motifs of the BnGRF family members, as well as the cis-acting regulatory elements in their promoters. We also analyzed the expression patterns of 15 randomly selected BnGRF genes in various tissues and in plant varieties with different harvest indices and gibberellic acid (GA) responses. The expression levels of BnGRFs under GA treatment suggested the presence of possible negative feedback regulation. The evolutionary patterns and expression profiles of BnGRFs uncovered in this study increase our understanding of the important roles played by these genes in oilseed rape.

[Effect of ketamine on the development of Chrysomya megacephala (Diptera : Calliphoridae)].

OBJECTIVE: To research the pattern of larvae and pupae development when exposed to ketamine. METHODS: The larvae of Chrysomya megacephala were reared in artificial diet containing ketamine with concentration of 0, 25, 50 and 100 mg/kg respectively at 32 degrees C, 28 degrees C and 24 degrees C in environmental chamber with a 12-h photoperiod and 75% humidity. 10 samples were collected from each group every 12 h from the 16th h after hatching to pupation. The max body length, weight of the larvae, growth rate of body length, weight and developmental duration of each stage were observed. RESULTS: The average length and weight in the treatment groups were significantly less than the control before achieving the maximum (P < 0.05), and the growth rate of 1/2LD50 group at 24 degrees C was most retarded. No dose dependence were observed among the ketamine fed groups. The effect of ketamine dose, temperature and their interaction on larval length and weight was statistically significant (P < 0.01). The effect of ketamine dose, temperature and their interaction account for, respectively, 20.9%, 60.2% and 18.9% of the total effect on growth of larval length, and they account for 8.3%, 85.6% and 6.1% of the total effect on growth of larval weight. The duration of larval stage in treatment groups was significantly delayed in comparison to the control at different temperatures (P < 0.05), and the duration of prepupal stage in treatment groups was significantly delayed (P < 0.05). However, the duration of pupal stage in treatment groups at 24 degrees C was significantly shorter than the control (P < 0.05). CONCLUSION: The time achieving maximum length and weight was significantly delayed, which results in an increased development duration of larval and prepupal stages, indicating that ketamine inhibits the growth of the larvae of C. megacephala.

[3 polymorphisms of gene GP IIb and the function of GP IIb T13959 G in the platelet transfusion refractoriness].

OBJECTIVE: To investigate the relationship among 3 polymorphisms of GP IIb and the function of GP IIb T13959 G in the platelet transfusion refractoriness(PTR). METHODS: The 26th exon, the 30th exon and the 21st intron of gene GP IIb in 110 individuals were amplified by polymerase chain reaction (PCR), and the PCR products were analyzed with single-strand conformation polymorphism(SSCP) and sequenced to investigate whether there was linkage among the polymorphisms of the gene. Human platelet antigen-3 (HPA-3) gene frequency was detected by Fok I enzyme in 147 patients with hematologic diseases, and was compared with that in 110 normal individuals. Forty-four patients who received apheresis platelet transfusion repeatedly were randomly divided into the HPA-3 homotype group and the control group. The antibodies of the platelet were detected after 3 times of platelet transfusion. RESULTS: There were polymorphisms of gene GP IIb in the 26th, 30th exon and the 21st intron, and the mutations were: T changed into G in 13,959 th of the 26th exon; C changed into T in 16,997 th of the 30th exon; the 9 bps deletion occurred in 11,996-12,004 th of the 21st intron. The 3 polymorphisms had synchronization in the individuals. The results of Fok I enzyme indicated that the frequency of HPA-3a was 83.6% (92/110)and 81.9%(119/147), and that of HPA-3b was 16.4%(18/110) and 19.1%(28/147) in the normal individuals and the patients respectively. There was no significant difference between the patients and normal individuals (P>0.05). After the platelet transfusion, the antibodies of all the cases of the homotype platelet transfusion were negative, but the antibodies in 2 cases of the control group were positive, and there was antibody to HPA-3a in one of the antibodies positive cases. CONCLUSION: (1)There is close linkage among the polymorphisms of gene GP IIb, which is T->G in 13 959 th of the 26th exon, C->T in 16,997 th of the 30th exon, and the 9 bps deletion in 11,996-12,004 th in the 21st intron. (2)The gene frequency of HPA-3a/3b is similar in the normal individuals and patients with hematologic diseases. (3) HPA-3 system may be one of the reasons for PTR in Chinese.

[Analysis of volatile components from the flowers of Chrysanthemum morifolium by GC-MS with solid-phase microextraction].

OBJECTIVE: To study a method for extraction and analysis of volatile components from Chrysanthemum morifolium 'gonghuangjv' cv. nov (CM GHJ) and C. morifolium 'gongbaijv' cv. nov (CM GBJ) by solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS). METHOD: The volatile components were extracted in different temperature, different balance period and different extraction fiber using head space solid-phase microextraction (HS-SPME), and were identified by CGC-MS. The variety in integral area of each component was observed in different conditions and its relative content was determined by normalization of area. RESULT: The better condition of SPME for C. morifolium was that the sample was extracted using 100 microm polydimethylsiloxane (PDMS) extraction fiber after it had been balanced for 6 hours at 75 degrees C. 53 components from CM GHJ and CM GBJ were identified, and there were 35 same components in CM GHJ and CM GBJ. CONCLUSION: HS-SPME-GC-MS is convenient, rapid and reliable for analysis of volatile components in C. morifolium.