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Background: Solid pseudopapillary neoplasms (SPNs) are uncommon tumors of low malignancy with a generally favorable prognosis, mostly originating from the pancreas. To date, 12 cases of SPNs with a primary ovarian origin (SPN-Os) have been reported globally, and their detailed characteristics have not been fully elucidated. Case description: We reported the 13th SPN-O case, which occurred in a 52-year-old woman with an 18.5 cm left ovarian mass. Four imaging methods, including ultrasound, computed tomography, magnetic resonance imaging and positron emission tomography, were utilized before surgery. An elevated level of serum cancer antigen 125 was detected and a total hysterectomy plus bilateral salpingo-oophorectomy was performed. Microscopic examination revealed a typical solid pseudopapillary structure. The tumor cells were stained focally for pan-cytokeratin, synaptophysin, CD99 and CD10, while ß-catenin, vimentin and CD56 were diffusely expressed. The Ki-67 proliferation index was 3%, and immunohistochemical (IHC) staining for chromogranin-A, inhibin-a, and E-cadherin was negative. No evidence of recurrence or metastasis was observed by clinical and imaging data during a 5-month postoperative follow-up. Conclusion: This is a report of an unusual case of a primary ovarian SPN with an up-to-date review of SPN-Os. A minimum combination of imaging methods and IHC stains was proposed for SPN-Os, which may prove beneficial in clinical practice.
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OBJECTIVE@#To investigate early effectiveness of arthroscopic superior fulcrum reconstruction in the treatment of irreparable massive rotator cuff tear (IMRCT).@*METHODS@#A retrospective analysis was conducted on the clinical data of 24 patients with IMRCT who met the inclusion criteria between January 2020 and April 2022. Among them, there were 11 males and 13 females with an average age of 56.2 years (range, 42-68 years). There were 12 cases of falling injuries, 3 cases of traction injuries, and the other 9 cases had no obvious causes. The disease duration ranged from 1 to 25 months (median, 6 months). The rotator cuff tears were classified as Hamada grade 2 in 18 cases and grade 3 in 6 cases, and Goutallier grade 1 in 3 cases, grade 2 in 20 cases, and grade 3 in 1 case. All patients were treated with arthroscopic superior fulcrum reconstruction. Visual analogue scale (VAS) score, Constant-Murley score, the University of California at Los Angeles (UCLA) score, and the American Shoulder and Elbow Surgeons (ASES) score were recorded before operation and at 1, 3, 6, and 12 months after operation.@*RESULTS@#The operations were all successfully completed. The incisions healed by first intention and no related complications occurred. All patients were followed up 12-33 months (mean, 24.6 months). The VAS, Constant-Murley, UCLA, and ASES scores at different time points after operation were superior to those before operation ( P<0.05). All of the above indicators further improved with time. Except for no significant difference in VAS and Constant-Murley scores between 6 and 12 months ( P>0.05), the differences between the other time points were significant ( P<0.05). At 12 months after operation, according to UCLA scoring standard, shoulder joint function was rated as excellent in 4 cases, good in 19 cases, and poor in 1 case, with an excellent and good rate of 96.0%. MRI showed that there was no graft re-tear and the transplanted tendon and bone tunnel healed.@*CONCLUSION@#The arthroscopic superior fulcrum reconstruction for IMRCT can effectively relieve the pain, improve the shoulder range of motion, and restore good shoulder function.
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Masculino , Femenino , Humanos , Persona de Mediana Edad , Lesiones del Manguito de los Rotadores/cirugía , Manguito de los Rotadores/cirugía , Estudios Retrospectivos , Resultado del Tratamiento , Artroscopía , Articulación del Hombro/cirugía , Rango del Movimiento ArticularRESUMEN
Granulomatous lobular mastitis (GLM) is a rare and chronic benign inflammatory disease of the breast. Difficulties exist in the management of GLM for many front-line surgeons and medical specialists who care for patients with inflammatory disorders of the breast. This consensus is summarized to establish evidence-based recommendations for the management of GLM. Literature was reviewed using PubMed from January 1, 1971 to July 31, 2020. Sixty-six international experienced multidisciplinary experts from 11 countries or regions were invited to review the evidence. Levels of evidence were determined using the American College of Physicians grading system, and recommendations were discussed until consensus. Experts discussed and concluded 30 recommendations on historical definitions, etiology and predisposing factors, diagnosis criteria, treatment, clinical stages, relapse and recurrence of GLM. GLM was recommended as a widely accepted definition. In addition, this consensus introduced a new clinical stages and management algorithm for GLM to provide individual treatment strategies. In conclusion, diagnosis of GLM depends on a combination of history, clinical manifestations, imaging examinations, laboratory examinations and pathology. The approach to treatment of GLM should be applied according to the different clinical stage of GLM. This evidence-based consensus would be valuable to assist front-line surgeons and medical specialists in the optimal management of GLM.
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Mastitis Granulomatosa , Mama/patología , Consenso , Femenino , Mastitis Granulomatosa/diagnóstico , Mastitis Granulomatosa/patología , Mastitis Granulomatosa/terapia , Humanos , RecurrenciaRESUMEN
Breast cancer is the most common malignant tumour in women. Our research on alloimperatorin from Angelica dahurica showed that alloimperatorin inhibited breast cancer cell viability in a concentration- and time-dependent manner; it also showed that apoptosis and ferroptosis inhibitors significantly weakened the antisurvival effect of alloimperatorin. Alloimperatorin clearly induced breast cancer cell apoptosis and increased the activities of caspase-3, caspase-8, caspase-9, and poly (ADP-ribose) polymerase; it also caused significant mitochondrial shrinkage, promoted the accumulation of Fe2+, reactive oxygen species, and malondialdehyde, and significantly reduced mRNA and protein expression levels of SLC7A11 and GPX4, indicating that alloimperatorin induces ferroptosis. In addition, alloimperatorin significantly promoted Kelch-like ECH-associated protein 1 (Keap1) expression; although it did not affect the expression of PGAM5 (mitochondrial serine/threonine protein phosphatase) and apoptosis-inducing factor mitochondria associated 1 (AIFM1), it significantly reduced the phosphorylation level of AIFM1. After downregulating the expression of Keap1, PGAM5, or AIFM1, the inhibitory effect of alloimperatorin on cell viability was significantly weakened, indicating that alloimperatorin regulates the Keap1/PGAM5/AIFM1 pathway to promote oxeiptosis. Alloimperatorin significantly inhibited the invasion of breast cancer cells, while Keap1 siRNA or GPX4 overexpression vectors significantly enhanced cell invasion and effectively reversed the anti-invasive effect of alloimperatorin. Therefore, alloimperatorin induces breast cancer cell apoptosis, ferroptosis, and oxeiptosis, thereby inhibiting cell growth and invasion.
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Neoplasias de la Mama , Ferroptosis , Apoptosis , Neoplasias de la Mama/patología , Femenino , Humanos , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Factor 2 Relacionado con NF-E2/genética , Poli(ADP-Ribosa) Polimerasas/metabolismoRESUMEN
The composition, physical and chemical properties, sources, and temporal and spatial changes in airborne particulate matter have been extensively investigated in previous studies. However, less is known about bioaerosols, which are mainly composed of bacteria and fungi and constitute up to 25% of the total airborne particulate matter. In this study, we used inductively coupled plasma mass spectrometry and ion chromatography to determine the concentrations of trace elements and water-soluble ions in atmospheric particulates, respectively. These analyses were combined with high-throughput sequencing methods and real-time quantitative polymerase chain reaction to analyze the microbial compositions in PM1.0, PM2.5, and PM10 samples, which were collected from July to September in Hefei City. The results showed that there were no significant differences in the bacterial community diversity across the three size fractions (analysis of variance (ANOVA), P>0.05). The bacterial and fungal community diversities on sunny days were lower than those on rainy days, and the bacterial community diversity in all samples was significantly higher than the fungal community diversity (ANOVA, P<0.01). The predominant bacterial phyla were Proteobacteria (46.19%), Firmicutes (33.42%), Bacteroidetes (10.99%), Cyanobacteria (3.33%), and Actinobacteria (2.11%). Ascomycota (73.23%), Basidiomycota (5.78%), Mortierellomycota (3.41%), and Mucoromycota (0.10%) were the dominant fungal phyla. Our results indicated that soils, plant leaves, and animal feces were the dominant sources of airborne bacterial communities in Hefei City, and the main sources of the fungal communities were plant leaves and soils. The bacterial community was mainly affected by K, Pb, Al, Fe, Mg, Ca, Na+, NO2-, and wind speed, and the main influencing factors of the fungal community were V, Mn, Sr, NO2-, NO3-, Na+, Cl-, the air quality index, and PM10. In addition, nine specific bacteria and fungi that are linked to human health risks were identified, including Acinetobacter, Streptococcus, Enterobacter, Pseudomonas, Delftia, Serratia, Trichoderma, Alternaria, and Aspergillus, which can lead to a wide range of diseases in humans and other organisms. The research results are helpful for revealing the various characteristics of airborne microbial communities, their influencing factors, and their impacts on human health, and are an important reference for subsequent research and the formulation of government policies.
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Contaminantes Atmosféricos , Microbiota , Microbiología del Aire , Contaminantes Atmosféricos/análisis , Animales , Ciudades , Monitoreo del Ambiente , Hongos/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Material Particulado/análisisRESUMEN
This retrospective analysis evaluated the interaction between Ki-67 and histological grade and their prognostic role in different breast cancer subtypes. In total, 2,573 breast cancer patients underwent surgery, and their histological grade and Ki-67 values were evaluated by breast pathologists. The median Ki-67 index was 15%, which was used as the cut-off for low/high Ki-67 expression. Recurrence-free survival (RFS) was calculated and compared, and the results indicated that Ki-67 expression was significantly associated with histological grade in all breast cancer patients (p < 0.001) and in each immunohistochemical (IHC)-based subtype (p < 0.001). Both high Ki-67 expression and grade 3 tumours were independent predictors of inferior RFS in all patients, especially in those with luminal-like tumours (p < 0.05). Ki-67 was an independent prognostic factor for RFS in grade 1, 2 patients with luminal-like tumours (adjusted hazard ratio [HR] = 1.92, 95% confidence interval [CI]: 1.22-3.03, p = 0.005), but not in the other subtypes. Similarly, histological grade predicted shorter RFS in patients with low Ki-67 expression who had luminal-like tumours (adjusted HR = 2.12, 95% CI: 1.13-3.99, p = 0.02) but not in the other subtypes. Conversely, Ki-67 showed no prognostic value for patients with grade 3 tumours and vice versa.
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Biomarcadores de Tumor , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Antígeno Ki-67/metabolismo , Adulto , Anciano , Neoplasias de la Mama/genética , Neoplasias de la Mama/mortalidad , Femenino , Expresión Génica , Humanos , Inmunohistoquímica , Estimación de Kaplan-Meier , Antígeno Ki-67/genética , Persona de Mediana Edad , Clasificación del Tumor , Estadificación de Neoplasias , Recurrencia , Factores de Riesgo , Adulto JovenRESUMEN
Since the introduction of ultra-low emissions, the characteristics of particulate matter (PM) emissions from coal-fired power plants have changed. We quantitatively evaluate the emission characteristics of each component in PM and the impact of purification equipment by analyzing three ultra-low emission units of coal-fired power plants (FP1, FP2, and FP3). A DGI was used to sample particles from the wet flue gas desulfurization (WFGD) unit and wet electrostatic precipitator (WESP) inlet and outlet, which were then analyzed by various methods. The results showed that the mass concentrations of PM1, PM2.5, and PM10 discharged from the outlets of the three units were 0.25-0.38, 0.31-0.42, and 0.42-0.57 mg·m-3, respectively, and that the mass concentration of PM10 discharged under the two kinds of units was equivalent. However, there were differences in the particle size distribution and composition of the particles. In comparison to the FP1 and FP2 units, the PM2.5/PM10 ratio of the FP3 unit was the highest. A possible reason for this is that the unit was equipped with a WESP, which can better remove particle sizes of 2.5 µm or more. The total concentrations of water-soluble ions in PM2.5 discharged from the FP2 and FP3 units were 0.20 and 0.06 mg·m-3, respectively. The water-soluble ions emitted from the FP2 unit were mainly Ca2+ and SO42-, whereas those mainly emitted from the FP3 unit were NH4+ and SO42-. Analysis of the PM from the WFGD import and export of the FP2 unit showed that the WFGD process increased the water-soluble ion discharge by entraining the desulfurization slurry containing limestone and gypsum. Addition of a WESP after WFGD can effectively remove PM2.5 and PM10 particles and reduce the influence of water-soluble ions on the atmospheric environment.
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A comparative analysis was conducted using total ozone products derived from monitoring atmospheric composition and climate (MACC) reanalysis data validated with ozone data from the atmospheric infrared sounder (AIRS) satellite and ground-based ozone measurements. The results indicate that the relative deviation of total ozone from the MACC reanalysis data and the ground-based ozone total data is controlled within 17%, and all of the correlation coefficients were between 0.79 and 0.97. The total ozone values from the MACC reanalysis data showed good consistency with the ground-based ozone measurements. With respect to the spatial distribution of multi-year averages, the relative deviation of total ozone values in the MACC reanalysis data and the AIRS satellite data was between -3% and 5%. The total ozone values in the MACC reanalysis data were higher than those from AIRS measurements for the Qinghai-Tibet Plateau and the coastal areas of South China, and were lower for northeast China. Furthermore, the seasonal variations in total ozone values in the MACC reanalysis data were consistent with AIRS data. At Mt. Waliguan station, the monthly averaged trends for near-surface ozone in the MACC reanalysis data were also consistent with surface ozone concentrations; the MACC reanalysis data reflect the observed trends for surface-based ozone measurements in spring, summer, and autumn, but show a large deviation in winter.
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A new benzylated alkamide, N-(3,4-dimethoxybenzyl)-9Z-oleamide (1), along with two known ones (2 and 3) were isolated from the roots of Lepidium meyenii collected from Lijiang, Yunnan Province of China. Their structures were elucidated by extensive spectroscopic analyses and the new compound further confirmed by a one-step synthesis. All the isolated alkamides were evaluated for their cytotoxicity against five human cancer cell lines. However, no significant activities were detected at concentrations up to 40 µM.
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Compuestos de Bencilo/aislamiento & purificación , Lepidium/química , Alcamidas Poliinsaturadas/aislamiento & purificación , Antineoplásicos Fitogénicos/síntesis química , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/farmacología , Compuestos de Bencilo/química , Compuestos de Bencilo/farmacología , Línea Celular Tumoral , China , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Espectroscopía de Resonancia Magnética , Estructura Molecular , Raíces de Plantas/química , Alcamidas Poliinsaturadas/química , Alcamidas Poliinsaturadas/farmacologíaRESUMEN
Objective To establish an acute yunaconitine poisoning rat model with a single oral administration and to determine the contents of yunaconitine in rat tissues by UPLC-MS/MS method, then investigate the distribution of yunaconitine in rats. Method The rats were randomly divided into three groups and were intragastrically administered a single dose of 2.2mg/kg,1.1mg/kg,0.7mg/kg yunaconitine, respectively.. The rats were killed 2h later, the stomach tissue, intestine tissue, liver tissue, pancreas tissue, kidney tissue, lung tissue, spleen tissue, heart tissue, bladder tissue, testis tissue, brain tissue and heart blood samples were collected. The contents of yunaconitine in the biological materials were determined by UPLC-MS/MS method after the biological samples extracted by liquid-liquid extraction. Result A rat model of the yunaconitine poisoning was made with a single dose of 1.1mg/kg, the concentrations of yunaconitine displayed in the organs with the following order:stomach, small intestine, liver, pancreas, kidney, lung, spleen, heart, bladder, testis, heart blood and brain. Conclusion Yunaconitine was widely distributed in rats, especially the levels in the stomach, small intestine and liver were the highest. The conclusion provides a basis for the selection of test materials for the poisoning of Aconitum vilmorinianum Kom.
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AIM: Ketamine and ethanol are increasingly being used together as recreational drugs in rave parties. Their effects on the dopamine (DA) system remain largely unknown. This study aimed to investigate the effects of consuming two different concentrations of ketamine with and without alcohol on the DA system. MATERIALS AND METHODS: We employed the conditioned place preference (CPP) paradigm to evaluate the rewarding effects of the combined administration of two different doses of ketamine (30mg/kg and 60mg/kg) with ethanol (0.3156g/kg). We evaluated the effects of the combined drug treatment on the transcriptional output of tyrosine hydroxylase (TH), dopa decarboxylase (DDC), synaptosomal-associated protein 25 (SNAP25), and vesicular monoamine transporter 2 (VMAT2) as well as protein expression level of brain-derived neurotrophic factor (BDNF) in rat prefrontal cortex (PFC) and striatum. KEY FINDINGS: We found that rats exhibited a dose-dependent, drug-paired, place preference to ketamine and ethanol associated with an elevated DA level in the striatum but not in the PFC. Moreover, treatment involving low- or high-dose ketamine with or without ethanol caused a differential regulatory response in the mRNA levels of the four DA metabolism genes and the cellular protein abundance of BDNF via the cortex-striatum circuitry. SIGNIFICANCE: This study investigated the molecular mechanisms that occur following the combined administration of ketamine and ethanol in the DA system, which could potentially lead to alterations in the mental status and behavior of ketamine/ethanol users. Our findings may aid the development of therapeutic strategies for substance abuse patients.
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Cuerpo Estriado/efectos de los fármacos , Dopamina/metabolismo , Etanol/farmacología , Ketamina/farmacología , Corteza Prefrontal/efectos de los fármacos , Animales , Conducta Animal/efectos de los fármacos , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Condicionamiento Psicológico/efectos de los fármacos , Cuerpo Estriado/metabolismo , Relación Dosis-Respuesta a Droga , Etanol/administración & dosificación , Ketamina/administración & dosificación , Masculino , Corteza Prefrontal/metabolismo , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
The distributions of size-segregated particles (PM10) and water-soluble ions (WSIs) in Taiyuan were studied from July 2014 to April 2015 by TE-235 aerosol sampling and ion chromatography analyzing. As the results shown, the daily PM10 level was 173.7 µg·m-3, which exceeded the Grade â ¡ limitation value in the Ambient Air Quality Standard (150 µg·m-3, GB 3095-2012). PM10 levels varied seasonally, and its were 199.1 and 194.2 µg·m-3 in winter and spring, respectively, which were much higher than those in summer. The PM10 size-segregated was bimodal distribution for the range of 0-0.95 and 3.0-7.2 µm. The concentration of WSIs was the highest in winter, followed by summer and spring. SO42-, NO3- and NH4+ were the main water soluble ions and accounted for 66% to 80% of the total WSIs. SO42-, K+, NH4+ and Cl- showed a unimodal distribution at <0.95 µm in all samples, while Ca2+ and Mg2+ showed a bimodal distribution at <0.95 µm and 3.0-7.2 µm. NO3- showed a unimodal size distribution at <0.95 µm in winter and spring, compared with a bimodal distribution at <0.95 µm and 3.0-7.2 µm in summer. By the correlation analysis, PM10 and WSIs decreased with the increase of wind speed in summer and winter other than in spring for the road-dust re-suspension by strong wind. Based on the ratio analysis of NO3-/SO42- and Mg2+/Ca2+, coal combustion was the main source of NO3- and SO42-, while Mg2+ and Ca2+ were mainly from the dust and coal combustion.
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Aerosoles/análisis , Contaminantes Atmosféricos/análisis , Monitoreo del Ambiente , Iones/análisis , China , Tamaño de la Partícula , Material Particulado/análisis , Estaciones del Año , Solubilidad , AguaRESUMEN
Precise quantification of Microthrix parvicella, which is identified as a dominated filamentous bacterium of bulking sludge in the worldwide, is essential for bulking investigation and related control strategies. However, quantitative processing based on fluorescence in situ hybridization (FISH) is prone to interference due to the specific characteristics of Microthrix parvicella (hydrophobic surface with thick cell wall). Our study focused on pretreatment and process optimization to show that the proportion of Microthrix parvicella was increased from 1.12% to 96.70% benefited by lysozyme (36000 U·mL-1), high probe concentration (4.5 ng·µL-1) and longer hybridization time (4 h) employed, mapping with the results of q-PCR method and Eikelboom & Jenkins Observation.
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Actinobacteria/aislamiento & purificación , Hibridación Fluorescente in Situ , Aguas del Alcantarillado/microbiología , Interacciones Hidrofóbicas e HidrofílicasRESUMEN
Three kinds of synthetic antibiotic (spiramycin, oxytetracycline, streptomycin) wastewater were continuously treated by parallel aerobic biofilm reactors for 6 months, respectively. Sludge bulking phenomenon caused by overgrowth of filamentous bacteria was observed under long-term high rbCOD and high C/N conditions in all reactors as showed by the Eikelboom and Jenkins examinations. The qualitative analysis of filamentous bacterial population in the biofilm and suspended sludge using fluorescence in situ hybridization (FISH) showed that the dominated filamentous bacteria in the bioreactor were N. limicola II and Thiothrix II. Under conditions of different antibiotic concentrations (5 mg x L(-1), 25 mg x L(-1)), there was no obvious change in the COD removal efficiency of the parallel reactors, while the NH4+ -N concentration (about 20 mg x L(-1)) occurred under high streptomycin concentration. The filamentous bacteria abundance was reduced with the increasing antibiotic concentration, especially significant for N. limicola II. Terramycin had a significant inhibitory effect on filamentous bacteria population, followed by Streptomycin and Spiramycin.
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Antibacterianos/química , Bacterias/efectos de los fármacos , Aguas del Alcantarillado/microbiología , Reactores Biológicos , Hibridación Fluorescente in Situ , Oxitetraciclina , Espiramicina , Estreptomicina , Aguas Residuales/microbiologíaRESUMEN
The contents of cholesterol in a number of vegetable oils were determined by saponification-gas chromatography ( FID) . There was quite a large possibility that cholesterol peak was seriously interfered by saponification-gas chromatography ( FID) , so it was difficult to set a cholesterol content value to differentiate vegetable oil from waste oil. Solid phase extraction ( SPE) sample pretreatment was chosen and the process conditions were optimized. The optimal conditions were as follows:0. 25 g of oil samples, 20 mL of 0. 6%ethyl ether-hexane ( V/V) as eluent ( get rid of fatty acid glycerides) and 20 mL of 15% ethyl ether-hexane ( V/V) as eluent ( obtain cholesterol) . An obvious cholesterol-content-difference between vegetable oils and waste oils was found by SPE-GC. The detected cholesterol contents in 84 vegetable oil samples were all less than 50 μg/g and the contents of 11 waste oil samples among the 13 waste oils were greater than 50 μg/g. Therefore, cholesterol content in vegetable oil of more than 50 μg/g could be determined as suspected waste oil, not vice versa by SPE-GC. The correlation coefficient R2 was 0. 9999 in 0-760 mg/L concentration range, and the detection limit was 6. 0 μg/g. Relative standard deviations at two concentration levels of 17. 7 and 695 mg/L were 1. 6% and 1. 5% respectively. The recovery was 103%.
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Leaf sheath color plays an important role as a marker for rice genetic improvement. A recombinant inbred line (RIL) population consisting of 220 individuals was developed from a cross between an Oryza sativa subsp. indica variety, IRBB60, and an Oryza sativa subsp. japonica variety, 9407. Within the RIL population, a line, RI51, was found to have purple leaf sheath (PSH). To map the gene governing PSH, RI51 was crossed with 9407 green leaf sheath (GSH) to develop an F2 segregating population. The distribution of F2 plants with PSH and GSH fitted a segregation ratio of 3:1, indicating that the PSH was controlled by a major dominant gene. The gene locus for PSH, tentatively designated as PSH1(t), was identified by surveying two bulks made of the respective 40 individuals with PSH and GSH with SSR markers covering the entire rice genome. The survey indicated that the PSH1(t) region was located on chromosome 1. Further confirmation was made using a large random sample of 360 individuals from the same F2 population and the PSH1(t) locus was then mapped on chromosome 1 between SSR markers RM3475 and RM7202 with genetic distances of 2.0 and 1.1 cM, respectively. For fine mapping of PSH1(t), a large F(2:3) segregating population with 3300 individuals from the seven heterozygous F2 plants in the RM3475-RM7202 region was constructed. Analysis of recombinants in the PSH1(t) region anchored the gene locus to an interval of 23.5 kb flanked by the left marker L03 and the right marker L05. Sequence analysis of this fragment predicted six open reading frames encoding a putative trans-sialidase, a putative Plastidic ATP/ADP-transporter, and four unknown proteins. The detailed genetic and physical maps of the PSH1(t) locus will be very useful in molecular cloning of the PSH1(t) gene.
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ADN de Plantas/genética , Genes de Plantas/genética , Oryza/genética , Hojas de la Planta/genética , Mapeo Cromosómico , Segregación Cromosómica , Cromosomas de las Plantas , Cruzamientos Genéticos , Genoma de Planta , Glicoproteínas/genética , Neuraminidasa/genética , Hojas de la Planta/crecimiento & desarrolloRESUMEN
A chitinase-encoding gene from Thermomyces lanuginosus was cloned by Rapid Amplification of cDNA Ends (RACE) using degenerated oligonucleotide primers designed from N-terminal amino acid sequence and the conserved amino acid sequence. The cloned full-length cDNA named chit, 1500bp in size, contained an OFR with 442 amino acids. The gene chit has been registered in GenBank with accession number DQ092332. The alignment results of putative amino acids sequence showed the catalytic domain of chit was high homology with the catalytic domains of the other chitinases in family-18, contained 2 conserved motifs related with catalytic activity of chitinase. The recombinant plasmid was generated by inserting the ORF of mature protein Chit into expression vector pPIC9K, and transformed into Pichia pastoris GS115. The recombinant chitinase was secreted successfully with the expression level of 0.36mg/mL and its activity could reach to 2.261U/mL after methanol induction 6d. The recombinant chitinase exhibited optimum catalytic activity at pH 5.5 and 60 degrees C. The enzyme was stable at 50 degrees C and the half life time at 65 degrees C was 40min.