Identification, Molecular Characteristics, and Evolution of YABBY Gene Family in Melastoma dodecandrum.

The YABBY gene family plays an important role in plant growth and development, such as response to abiotic stress and lateral organ development. YABBY TFs are well studied in numerous plant species, but no study has performed a genome-wide investigation of the YABBY gene family in Melastoma dodecandrum. Therefore, a genome-wide comparative analysis of the YABBY gene family was performed to study their sequence structures, cis-acting elements, phylogenetics, expression, chromosome locations, collinearity analysis, protein interaction, and subcellular localization analysis. A total of nine YABBY genes were found, and they were further divided into four subgroups based on the phylogenetic tree. The genes in the same clade of phylogenetic tree had the same structure. The cis-element analysis showed that MdYABBY genes were involved in various biological processes, such as cell cycle regulation, meristem expression, responses to low temperature, and hormone signaling. MdYABBYs were unevenly distributed on chromosomes. The transcriptomic data and real-time reverse transcription quantitative PCR (RT-qPCR) expression pattern analyses showed that MdYABBY genes were involved in organ development and differentiation of M. dodecandrum, and some MdYABBYs in the subfamily may have function differentiation. The RT-qPCR analysis showed high expression of flower bud and medium flower. Moreover, all MdYABBYs were localized in the nucleus. Therefore, this study provides a theoretical basis for the functional analysis of YABBY genes in M. dodecandrum.

[Serological Characteristics and Molecular Biological Mechanism of AEL.02 Subtype].

OBJECTIVE: To explore the serological characteristics and molecular biological mechanism of an ael subtype specimen. METHODS: The ABO blood typing was identified by routine blood group serological and absorption/elution methods; PCR-SBT method for ABO genotyping: 7 exons of ABO gene were amplified by PCR, the amplified products were purified, and then sequencing primers were designed and the amplified products were sequenced directly for analysis; 3D molecular model was constructed and the difference of free energy (ΔΔG) was used to predict the GTA mutant stability. RESULTS: A antigen was not detected on erythrocytes through absorption and elution tests, which was not consistent with the serological characteristics of ael, and the serological typing results were ambiguous. The ABO genotype was ABO*AEL.02/O.01.01, and there were two mutations in exon 7 of the gene, c.467C>T and c.646T>A, which could lead to the replacement of proline with leucine at position 156 (p.Pro156Leu) and phenylalanine with isoleucine at position 216 on the GTA, respectively. The 3D model predicts that the mutations do not introduce new hydrogen bonds to the GTA mutant and do not form a new secondary structure, but can lead to an increase in the ΔΔG value of the GTA mutant, suggesting a decrease in protein stability. CONCLUSION: The serological characteristics alone is not reliable to determine the ael subype; the ael phenotype may be due to the GTA mutant that reduces enzyme stability.

The Melastoma dodecandrum genome and the evolution of Myrtales.

Melastomataceae has abundant morphological diversity with high economic and ornamental merit in Myrtales. The phylogenetic position of Myrtales is still contested. Here, we report the chromosome-level genome assembly of Melastoma dodecandrum in Melastomataceae. The assembled genome size is 299.81 Mb with a contig N50 value of 3.00 Mb. Genome evolution analysis indicated that M. dodecandrum, Eucalyptus grandis, and Punica granatum were clustered into a clade of Myrtales and formed a sister group with the ancestor of fabids and malvids. We found that M. dodecandrum experienced four whole-genome polyploidization events: the ancient event was shared with most eudicots, one event was shared with Myrtales, and the other two events were unique to M. dodecandrum. Moreover, we identified MADS-box genes and found that the AP1-like genes expanded, and AP3-like genes might have undergone subfunctionalization. The SUAR63-like genes and AG-like genes showed different expression patterns in stamens, which may be associated with heteranthery. In addition, we found that LAZY1-like genes were involved in the negative regulation of stem branching development, which may be related to its creeping features. Our study sheds new light on the evolution of Melastomataceae and Myrtales, which provides a comprehensive genetic resource for future research.

BCL11A Promotes the Progression of Laryngeal Squamous Cell Carcinoma.

Background: We report functional and clinical data uncovering the significance of B-cell lymphoma/leukemia 11A (BCL11A) in laryngeal squamous cell carcinoma (LSCC). Methods: We examined BCL11A expression in a cohort of LSCC patients and evaluated the association between BCL11A expression and clinicopathological features. We investigated the consequences of overexpressing BCL11A in the LSCC cell line on proliferation, migration, invasion, cell cycle, chemosensitivity, and growth in vivo. We explored the relationship between BCL11A and MDM2 in LSCC and tumorigenesis pathways by using the Human Cancer PathwayFinder Array. Results: High levels of BCL11A were found in LSCC tissues and were more frequently associated with advanced lymphatic metastasis stages with poor prognoses. BCL11A overexpression enhanced LSCC proliferation in vitro and vivo. A positive correlation between MDM2 and BCL11A expression was identified. Conclusions: These data uncover important functions of BCL11A in LSCC and identify BCL11A as a prognostic biomarker and potential therapeutic target in LSCC.

[Retrospective analysis of 188 cases of parapharyngeal space tumors].

Objective:To explore the diagnosis,treatment,surgical approach and prognosis of parapharyngeal space tumors.Method:The clinical data of 188 patients with parapharyngeal space tumor who were treated from January 2007 to December 2016 were analyzed retrospectively.All patients underwent imaging examination before operation.Surgical approach was as follows:transcervical approach applied in 159 cases,endoscopic-assisted transnasal approach in 9 cases,transcervical-transmandibular approach in 8 cases,transcervical-transparotid approach in 8 cases,transoral approach in 7 cases,and infratemporal fossa approach in 4 case.Result:Of the 188 cases,the tumor was benign in nature in 168 cases(89%)and malignant in 20 cases(11%).Complications occurred in 28(15%)patients,with the most common symptom being hoarseness.168 cases of benign tumors were followed up for 10 months to 10 years,and 3 cases were lost and 4 cases had recurrence.All cases underwent re-operation.Patients with malignant tumors received combined treatment after surgery,and 3 cases were lost to follow-up,1 case died of recurrence 9 months after surgery,the rest survived.Conclusion:Surgery is the preferred method for treating parapharyngeal space tumors and postoperative recurrence rate is pretty low.Endoscopy provides a new surgical management method,helping to reduce postoperative complications and recurrence rate.

Kinetic resolution of racemic styrene oxide at a high concentration by recombinant Aspergillus usamii epoxide hydrolase in an n-hexanol/buffer biphasic system.

Using the cell-free extract of engineered E. coli/Aueh2, expressing the recombinant Aspergillus usamii epoxide hydrolase (reAuEH2), as a biocatalyst, the kinetic resolution technique of racemic styrene oxide (rac-SO) was examined. In a phosphate buffer system (50mM, pH 7.0), 200mM rac-SO was efficiently resolved, obtaining (S)-SO with 98.1% enantiomeric excess (e.e.), whereas (S)-SO only with 45.2% e.e. was obtained from 750mM rac-SO. The analytical results verified that reAuEH2 shows tolerance towards high substrate concentration but is inactivated at a product concentration of 300mM. To produce (S)-SO with the high concentration, e.e. and volumetric productivity, n-hexanol was selected from a variety of water-miscible and water-immiscible organic solvents to construct an n-hexanol/buffer biphasic system. The optimal phase volume ratio, substrate over enzyme ratio and temperature were 1:1 (v/v), 6:1 (w/w) and 25°C, respectively. In an optimized biocatalytic system, a gram-scale resolution of rac-SO at a high concentration of 1M (120g/L) was performed at 25°C for 2h, obtaining (S)-SO with 98.2% e.e., 34.3% yield (maximum yield of 50%). The substrate concentration and volumetric productivity (1M, 20.6g/L/h) in a biphasic system significantly increased compared with those (0.2M, 3.1g/L/h) in a phosphate buffer system. The efficient resolution of rac-SO at a high concentration in a biphasic system makes it a promising technique for preparing a highly value-added enantiopure (S)-SO with high volumetric productivity.

Association of the microsatellite (GT)n repeat polymorphisms of the HO-1 gene promoter and corresponding serum levels with the risk of laryngeal squamous cell carcinoma.

CONCLUSION: Long (GT)n repeat polymorphisms in the heme oxygenase-1 (HO-1) gene promoter and decreased serum HO-1 levels are associated with a higher susceptibility to laryngeal squamous cell carcinoma (LSCC). OBJECTIVE: In this case-control study, the association of HO-1 microsatellite (GT)n repeat polymorphisms and serum levels with the risk of LSCC was investigated. METHODS: A total of 142 LSCC patients, 54 vocal leukoplakia patients and 98 healthy controls, were examined for (GT)n polymorphisms by sequencing, and the serum HO-1 levels were detected in a sub-set from participants above by ELISA. RESULTS: Compared with the controls, the LSCC group had significantly higher frequencies of L-allele (> 29 repeats) and L-allele carriers (p < 0.001, OR = 2.037 and p = 0.005, OR = 2.152, respectively). The frequencies of lymph node metastasis and of moderate or poor differentiation were significantly higher in L-allele carriers compared to non-L-allele carriers (p < 0.05). Significantly lower serum HO-1 levels were detected in LSCC patients (p < 0.001), and patients with lower serum HO-1 levels had more advanced cancer stage and a higher lymph node metastasis rate (p < 0.05). Furthermore, the L-allele carriers had lower serum HO-1 concentrations compared with the non-L-allele carriers (p = 0.019).

[Investigation on pinworm infection and relative factors on prevalence among urban and rural preschool children in Xianyang City].

A total of Eight hundred eighty-six children from 3 to 7 years old in 8 kindergartens were sampled in urban and rural area in Xianyang City from March to May 2012. The cellophane tape swab technique was used to examine pinworm eggs. Children's hygiene habits, clinical symptoms and hygienic condition were surveyed by questionnairing. The total infection rate of pinworm was 11.2% (99/886). The rate in males and females was 10.4% (52/500) and 12.2% (47/386), respectively. The infection rate in rural kindergartens (19.1%, 70/367) was higher than that of urban kindergartens (5.6%, 29/519) (chi2 = 39.39, P < 0.01). Among the investigated children aged 3-7 years, the infection rate in 4-5 years group (12.7%) was the highest, but no statistical difference was found among age groups (P> 0.05). Multivariate logistic regression analysis showed that the hygiene habits such as washing hands before eating (OR = 0.180), drinking unboiled water and eating non-cooked food (OR = 2.473), cleaning perianal region frequently (OR = 0.836), cutting nails frequently (OR = 0.450), drying the quilt regularly (OR = 0.224) and health education (OR = 0.639) were the influence factors on pinworm infection. The main symptoms of pinworm infection include pruritus and bruxism.

[Changes in cardiac specific microRNA-208a level in peripheral blood in ST segment elevation acute myocardial infarction patients].

OBJECTIVE: To observe serum cardiac specific microRNA-208a (miR-208a) levels in ST segment elevation acute myocardial infarction (STEAMI) patients, and to explore the role of serum miR-208a levels in the diagnosis of STEAMI. METHODS: The serum miR-208a concentrations were assessed within 12 hours after STEAMI, while 30 healthy individuals as control. Serum miR-208a concentrations were measured with real-time quantity reverse transcription-polymerase chain reaction (qRT-PCR), and serum cardiac troponin I (cTnI) or MB isoenzyme of creatine kinase (CK-MB) concentrations were measured with enzyme linked immunosorbent assay (ELISA). RESULTS: The contents of serum cTnI or CK-MB in STEAMI patients were significantly higher than those in healthy individuals (cTnI: 17.72±8.43 µg/L vs. 0.05±0.01 µg/L, CK-MB: 250.83±177.26 µg/L vs. 71.20±20.50 µg/L, both P<0.01). Serum miR-208a concentrations were detected in all individuals with STEAMI within 60 PCR cycle (0-6 hours: 44.95±4.77, 6-12 hours: 43.98±4.68), but were beyond detection for all individuals in the healthy control group. The serum miR-208a relative levels in STEAMI patients were at least more than 215 fold than that in healthy persons, compared with qRT-PCR (Ct=60) of miR-208a in healthy control persons (P<0.01). CONCLUSION: Serum miR-208a may be a new biomarker the early diagnosis of STEAMI patients.

Synthesis and enhanced ethanol sensing characteristics of alpha-Fe2O3/SnO2 core-shell nanorods.

Alpha-Fe(2)O(3)/SnO(2) core-shell nanorods are synthesized via a three-step process. X-ray diffraction (XRD), scanning electron microscopy (SEM) and transmission electron microscopy (TEM) analyses reveal that their diameters and lengths are respectively in the ranges 35-120 nm and 0.35-1.2 microm, and the thickness of the shell composed of 3.5 nm SnO(2) nanoparticles is about 10 nm. The core-shell nanostructures exhibit a dramatic improvement in ethanol sensing characteristics compared to pure alpha-Fe(2)O(3) nanorods. The sensor response is up to 19.6 under 10 ppm ethanol exposure at 220 degrees C. Both the response time and the recovery time of the core-shell structures are less than 30 s. Based on the space-charge layer model and semiconductor heterojunction theory, the small thickness of the SnO(2) shell and the formation of heterojunctions contribute to the enhanced ethanol sensing characteristics. Our results demonstrate that one-dimensional metal oxide core-shell nanostructures whose shell thickness is smaller than the Debye length are very promising materials for fabricating gas sensors with good performances.

Combustion oxidization synthesis of unique cage-like nanotetrapod ZnO and its optical property.

Cage-like nano-tetrapod ZnO is a novel structure, which was successfully synthesized by combustion oxidation at 850 degrees C. No catalyst or carrier gases were used. Thorough SEM and TEM analyses revealed that the linking legs of the tetrapod ZnO can have or not interface. The formation or not of an interface is discussed and it was attributed to different growth process of the cage-like ZnO nano-tetrapod. Enhanced UV emission peak at the wavelength of 375 nm, featuring high intensity and narrow width, indicates a highly crystalline structure. A green emission, recorded at 502 nm, was related to the defects of the surface of the branching configuration as well as to the ZnO nuclei of the cage-like nano-tetrapod ZnO.

[Construction, expression and immunogenicity study of a chimeric MS/hIL-12 eukaryotic expression plasmid].

OBJECTIVE: To construct and express a chimeric Mtb8.4 with signal peptide (MS)/hIL12 eukaryotic expression plasmid, and to study the immunogenicity of the MS/hIL-12 chimeric genetic vaccines. METHODS: The MS/hIL-12 chimeric gene was amplified by polymerase chain reaction (PCR) and cloned into the eukaryotic expression vector pCI-neo. The correct pCI-neo-MS/hIL12 (pMSI) recombinant plasmid was identified by PCR, restricted enzyme digestion and DNA sequencing. COS-7 cells were transfected with pMSI constructs by cationic liposome. After 48 hours, mRNA of the target gene was detected by RT-PCR, and hIL-12 protein in culture supernatant and cell lysates was detected by Western blot. C57BL/6N mice were vaccinated with MS/hIL-12 chimeric gene vaccine for three times at 3 week intervals. Four weeks after the final inoculation, three mice were sacrificed for measurement of the cytokine response and cytotoxic T lymphocyte (CTL) induction. RESULTS: The accuracy of plasmid construction was confirmed by a number of molecular biological techniques. Transfection of COS-7 cells with plasmids pMSI lead to transient expression of fusion proteins. The IFN-gamma and IL-2 titers were (1,521 +/- 48) ng/L and (755 +/- 41) ng/L in MS/hIL-12 chimeric gene vaccine group, (820 +/- 50) ng/L and (297 +/- 31) ng/L in MS gene vaccine group, (1,487 +/- 40) ng/L and (767 +/- 50) ng/L in BCG group, (121 +/- 16) ng/L and (62 +/- 10) ng/L in vacant vector group, and (48 +/- 16) ng/L and (32 +/- 17) ng/L in PBS group respectively. The levels of IFN-gamma and IL-2 in MS/hIL-12 chimeric gene vaccine group were higher than those of MS gene vaccine group, vacant vector group and PBS group (P < 0.01) and was similar to the BCG group (P > 0.05). The level of IL-4 in BCG group [(91 +/- 11) ng/L] increased significantly as compared to other groups (P < 0.01). When effector-cell-to-target-cell ratio (E:T ratio) were 100:1, 50:1, and 10:1 respectively, the CTL activity was 77.5%, 51.2%, 30.3% in MS/hIL-12 chimeric gene vaccine group, 56.2%, 37.8%, 11.5% in MS gene vaccine group, 28.9%, 21.4%, 9.8% in BCG group. The cytotoxicity in MS/hIL-12 chimeric gene vaccine group was higher than that of other groups (P < 0.01). CONCLUSION: When used to construct the chimeric gene vaccine, hIL-12 could improve the immunogenicity of MS gene vaccine.

Promoter activity of SARS coronavirus 5' UTR sequence in eukaryotic cells.

OBJECTIVES: To investigate 5'UTR sequence in different SARS-CoV isolates, to identify the secondary structure, and to test the promoter activity of the cDNA sequence corresponding to SARS-CoV 5'UTR in eukaryotic cells. METHODS: 101 SARS-CoV 5'UTR were aligned. One typical sequence containing full 264 nt was then subjected to be predicted its secondary structure. The pGL3-5'UTR and pGL3-a-5'UTR were constructed by substitution of SV40 promoter with SARS-CoV 5'UTR cDNA or its antisense sequence. Then the recombinant plasmids were transfected into HepG2 cells and the luciferase activities were detected. A set of deletion mutant plasmids, of which pGL3-5' UTR-1, pGL3-5' UTR-2, pGL3-5'UTR-3 and pGL3-5'UTR-4 are with 3, 2, 1, and 0 residual stem-loops of 3' termini respectively,were constructed from pGL3-5'UTR and were transfected into HepG2 cells to express reporter gene luc+, with pGL3-5'UTR containing full sequence as control. The luciferase activities expressed by the plasmids were measured. And then the total RNA of the transfected cells was extracted. Subsequently, by 5' Rapid Amplication of cDNA Ends (5'RACE), the PCR product was sequenced. The luciferase expressed by pGL3-5'UTR in various cells, the lung carcinoma cell line A549, hepatoma cell line HepG2, kidney cell Vero E6, cervical cancer cell line HeLa and human umbilical vein endothelial cell line ECV304 were measured and compared with each other. RESULTS: The full sequence of the SARS-CoV 5' UTR is a 264nt, and 18 deletion mutants were found. Totally, 5 site substitutions were found in 101 5'UTR sequences. The SARS-CoV 5'UTR RNA folded to form a stable secondary structure containing four stem-loop domains. The biggest and most complex one is the stem-loop II appearing a pseudoknot. Comparing with pGL3-a-5'UTR, pGL3-5'UTR expressed luciferase obviously. Both pGL3-5'UTR containing full sequence and pGL3-5'UTR-1 containing three stem-loops of 3' termini expressed the luciferase well. However, when lost stem-loop I and II , the pGL3-5'UTR-2, pGL3-5'UTR-3 and pGL3-5'UTR-4 almost didn't express luciferase. The 56th nucleotide of SARS-CoV 5'UTR was found to be the initiation site for transcription. Transfected with expression luciferase plasmid pGL3-5' UTR in which SARS-CoV 5' UTR acts as the promoter, the luciferase could express in five cell lines in different degrees. Ranked by the luciferase activity from the highest to the lowest, the order is A549, HepG2, ECV304, HeLa and Vero E6. CONCLUSIONS: A: The 5'UTR sequences of different SARS-CoV isolates are relatively conserved, and a full sequence would form a secondary structure containing four stem-loop domains. B: The cDNA sequence corresponding to SARS-CoV 5'UTR possessed a promoter activity in eukaryotic cells. C: The promoter domain of the SARS-CoV 5'UTR contains both stem-loop I and II. D: The 56th nucleotide and its down stream TRS of SARS-CoV 5'UTR plays a key role in regulating transcription. E: Cells sourced from various tissues can provide efficient accessory factors for SARS-CoV 5'UTR sequence that acts as a promoter, and the lung-sourced cells may be the most suitable.