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1.
China Pharmacy ; (12): 2121-2127, 2023.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-987142

ABSTRACT

OBJECTIVE To explore the mechanism of Kuaisong yin in the prevention and treatment of constipation. METHODS Slow transit constipation (STC) model was established with Compound difenoxylate tablet in mice and rats. Two batches of mice were divided into blank group, model group, positive control group (Maren soft capsule, 0.64 g/kg), Kuaisong yin low-dose, medium-dose and high-dose groups (3.2, 6.4, 12.8 g/kg), with 10 mice in each group. The effect of Kuaisong yin on constipation in mice was evaluated by intestinal propulsion experiment and defecation experiment. Rats were divided into blank group, model group, positive control group (Maren soft capsule,0.36 g/kg), Kuaisong yin low-dose and high-dose groups (2.4, 4.8 g/kg), with 7 or 8 rats in each group. They were given relevant medicine once a day for 1 week. The metabonomics of serum and urine of rats were analyzed by UPLC-Q-TOF-MS/MS technology. RESULTS Compared with model group, the ink propulsion rate and 5 h defecation volume of mice in Kuaisong yin high-dose group were significantly increased (P<0.05); the first defecation time of mice in Kuaisong yin medium-dose and high-dose groups was significantly shortened, and the quality of defecation was significantly reduced within 5 h (P<0.05 or P<0.01). Serum metabonomics screened 16 compounds (such as proline, propionylcarnitine, hemolytic phosphatidylcholine, etc.) and 6 metabolic pathways (such as sphingomyelin metabolism, arginine and proline metabolism, sphingolipid biosynthesis-lactose and neolactone series). Urine metabonomics screened 20 different metabolites (such as prostaglandin A2, L-valine, phosphatidylcholine, sphingomyelin, etc.) and 8 metabolic pathways (such as valine, leucine and isoleucine biosynthesis, sphingomyelin metabolism, pyruvate metabolism, etc.). CONCLUSIONS Kuaisong yin can play a role in improving constipation by regulating different metabolites such as hemolytic phosphatidylcholine, phosphatidylcholine, prostaglandin A2, L-valine, proline, and regulating metabolic pathways such as multiple amino acid metabolism, sphingomyelin metabolism, etc.

2.
China Pharmacy ; (12): 1093-1098, 2023.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-972953

ABSTRACT

OBJECTIVE To study the mechanism of Compound zaoren granule in improving insomnia. METHODS Forty-nine mice were divided into blank group, model group, positive control group 1 (Estazolam tablets 0.5 mg/kg),control group 2 (Shumian capsule 0.6 g/kg), Compound zaoren granule low-dose, medium-dose and high-dose groups (2.5, 5, 10 g/kg), with 7 mice in each group. The insomnia model was established by chronic unpredictable mild stress combined with 4-chloro-DL- phenylacetic acid. The behavioral changes of mice were investigated through open field test and pentobarbital sodium synergistic hypnosis experiment, as well as the pathomorphology of mice hypothalamus tissue was observed by HE staining. The metabonomics analysis and multivariate statistical analysis of serum in mice were performed by UHPLC-Q-TOF-MS/MS, and the differential metabolites were screened out; the metabolic pathway analysis was conducted based on MetaboAnalyst 5.0 database. RESULTS Compared with blank group, the total travelling distance, the number of entering the central region and the moving distance in the central region of the model group were significantly reduced (P<0.05), the proportion of total rest time was significantly increased (P<0.05), the sleep duration of mice was significantly shortened (P<0.05), and hypothalamic nerve cells damaged and severely vacuolated. Compared with model group, the total travelling distance of Compound zaoren granule low-dose and medium-dose groups were increased significantly and the proportions of total rest time of those groups were decreased significantly (P<0.05), and the sleep duration of mice in Compound zaoren granule high-dose group was prolonged significantly (P<0.05); the hypothalamic nerve cells of mice in each administration group recovered to varying degrees, and the hypothalamus histiocytes of mice in the Compound zaoren granules high-dose group were closer to those in the blank group. A total of 18 differential metabolites (such as phenylalanine, taurine, norvaline, methionine) and 4 important amino acid metabolic pathways (L-phenylalanine, tyrosine and tryptophan biosynthesis; taurine and hypotaurine metabolism; L-phenylalanine metabolism; cysteine and methionine metabolism) were identified through metabolomics analysis. CONCLUSIONS Compound zaoren granules can normalize the disordered metabolism in vivo by regulating differential metabolites such as phenylalanine, taurine, and four amino acid metabolic pathways, so as to improve insomnia.

3.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-981811

ABSTRACT

OBJECTIVE@#To explore the clinical characteristics and genetic basis of a child with autism spectrum disorder (ASD) in conjunct with congenital heart disease (CHD).@*METHODS@#A child who was hospitalized at the Third People's Hospital of Chengdu on April 13, 2021 was selected as the study subject. Clinical data of the child were collected. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing (WES). A GTX genetic analysis system was used to analyze the WES data and screen candidate variants for ASD. Candidate variant was verified by Sanger sequencing and bioinformatics analysis. Real-time fluorescent quantitative PCR (qPCR) was carried out to compare the expression of mRNA of the NSD1 gene between this child and 3 healthy controls and 5 other children with ASD.@*RESULTS@#The patient, an 8-year-old male, has manifested with ASD, mental retardation and CHD. WES analysis revealed that he has harbored a heterozygous c.3385+2T>C variant in the NSD1 gene, which may affect the function of its protein product. Sanger sequencing showed that neither of his parent has carried the same variant. By bioinformatic analysis, the variant has not been recorded in the ESP, 1000 Genomes and ExAC databases. Analysis with Mutation Taster online software indicated it to be disease causing. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was predicted to be pathogenic. By qPCR analysis, the expression level of mRNA of the NSD1 gene in this child and 5 other children with ASD was significantly lower than that of the healthy controls (P < 0.001).@*CONCLUSION@#The c.3385+2T>C variant of the NSD1 gene can significantly reduce its expression, which may predispose to ASD. Above finding has enriched the mutational spectrum the NSD1 gene.


Subject(s)
Male , Child , Humans , Autism Spectrum Disorder/genetics , Heart Defects, Congenital/genetics , Computational Biology , Genomics , Mutation , RNA, Messenger/genetics , Histone-Lysine N-Methyltransferase/genetics
4.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-990545

ABSTRACT

The anatomical abnormalities associated with pancreatitis mainly include pancreas anomalies, pancreaticobiliary maljunction and intestinal duplication.Pancreas anomalies are the most common congenital abnormalities in the triggers of pancreatitis, including pancreas divisum, annular pancreas and heterotopic pancreas prevail.In all these anomalies, the mechanism of pancreatitis is likely due to outlet obstruction.Awareness of these anomalies is necessary to arrange the proper strategy for the treatment of patients with pancreatitis.

5.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-957530

ABSTRACT

Objective:To evaluate the relationship between autophagy and oxidative stress during remifentanil-induced hyperalgesia in the rats with incisional pain.Methods:Thirty-two clean-grade healthy male Sprague-Dawley rats, weighing 230-250 g, aged 2-3 months, were divided into 4 groups ( n=8 each) using a random number table method: incisional pain group (I group), remifentanil + incisional pain group (RI group), autophagy inhibitor 3-methyladenine + remifentanil + incisional pain group (MRI group) and autophagy agonist rapamycin group + remifentanil + incisional pain group (RRI group). The model of incision pain was developed and the equal volume of normal saline was intravenously infused simultaneously for 60 min.In RI, MRI and RRI groups, the model of incision pain was developed and remifentanil 1 μg·kg -1·min -1 was simultaneously infused for 60 min.In MRI group, 3-methyladenine 15 mg/kg was intraperitoneally injected at 12 h before developing the model, and rapamycin 10 mg/kg was intraperitoneally injected at 12 h before developing the model in RRI group.Thermal paw withdrawal latency (TWL) and mechanical paw withdrawal threshold (MWT) were measured at 24 h before infusion of remifentanil or normal saline (T 0) and at 2, 6, 24 and 48 h after the end of infusion (T 1-4). The rats were sacrificed after the end of behavioral testing, and the lumbar enlargement segments of the spinal cord were harvested for determination of the expression of autophagy microtubule-associated protein 1 light chain 3Ⅱ (LC3Ⅱ), Beclin-1 and P62 (by Western blot), activity of superoxide dismutase (SOD) (by xanthine oxidase method) and content of malondialdehyde (MDA) (by thiobarbital method). Results:Compared with the baseline at T 0, PWT was significantly decreased and TWL was shortened at T 1-4 in the four groups ( P<0.05). Compared with I group, MWT was significantly decreased and TWL was shortened at T 1-4, the expression of LC3 Ⅱand Beclin-1 was up-regulated, the expression of P62 was down-regulated, SOD activity was decreased, and MDA content was increased in RI group ( P<0.05). Compared with RI group, MWT was significantly decreased and TWL was shortened at T 1-4, the expression of LC3 Ⅱand Beclin-1 was down-regulated, the expression of P62 was up-regulated, SOD activity was decreased, and MDA content was increased in MRI group ( P<0.05), and MWT was significantly increased and TWL was prolonged at T 1-4, the expression of LC3 Ⅱand Beclin-1 was up-regulated, the expression of P62 was down-regulated, SOD activity was increased, and MDA content was decreased in RRI group ( P<0.05). Conclusions:Autophagy is involved in the process of remifentanil-induced incisional hyperalgesia through regulating the level of oxidative stress in rats.

6.
Chinese Journal of Biotechnology ; (12): 737-748, 2022.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-927740

ABSTRACT

CRISPR/Cas9 has been widely used in engineering Saccharomyces cerevisiae for gene insertion, replacement and deletion due to its simplicity and high efficiency. The selectable markers of CRISPR/Cas9 systems are particularly useful for genome editing and Cas9-plasmids removing in yeast. In our previous research, GAL80 gene has been deleted by the plasmid pML104-mediated CRISPR/Cas9 system in an engineered yeast, in order to eliminate the requirement of galactose supplementation for induction. The maximum artemisinic acid production by engineered S. cerevisiae 1211-2 (740 mg/L) was comparable to that of the parental strain 1211 without galactose induction. Unfortunately, S. cerevisiae 1211-2 was inefficient in the utilization of the carbon source ethanol in the subsequent 50 L pilot fermentation experiment. The artemisinic acid yield in the engineered S. cerevisiae 1211-2 was only 20%-25% compared with that of S. cerevisiae 1211. The mutation of the selection marker URA3 was supposed to affect the growth and artemisinic acid production. A ura3 mutant was successfully restored by a recombinant plasmid pML104-KanMx4-u along with a 90 bp donor DNA, resulting in S. cerevisiae 1211-3. This mutant could grow normally in a fed-batch fermentor with mixed glucose and ethanol feeding, and the final artemisinic acid yield (> 20 g/L) was comparable to that of the parental strain S. cerevisiae 1211. In this study, an engineered yeast strain producing artemisinic acid without galactose induction was obtained. More importantly, it was the first report showing that the auxotrophic marker URA3 significantly affected artemisinic acid production in a pilot-scale fermentation with ethanol feeding, which provides a reference for the production of other natural products in yeast chassis.


Subject(s)
Artemisinins , Fermentation , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/metabolism
7.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-989288

ABSTRACT

Sarcopenia is a common clinical syndrome in cancer patients and is becoming an independent poor prognostic factor. Studies have shown that an appropriate increase in protein, vitamin D, Omega-3, and other nutrient intake combined with exercise, while ensuring appropriate energy intake, can improve or even reverse sarcopenia. In this review, the research progress in the assessment and diagnosis, epidemiological features, clinical outcomes, and interventions for sarcopenia in patients with head and neck cancer was reviewed.

8.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-994536

ABSTRACT

Objective:To analyze the causes of unplanned reoperation in pediatric patients after elective digestive tract surgery and the prognosis.Methods:Medical records were reviewed from pediatric patients undergoing unplanned reoperation after elective digestive tract surgery at our department from Jan 2012 to Dec 2019. Primary diagnoses, procedures and levels of index surgeries, causes and procedures of unplanned reoperations, and patients' prognosis were analyzed.Results:There were 39 cases, and the primary diagnoses included biliary disease, anal and colorectal disease, and intestinal disease. There were 4 (10%) cases of level Ⅱ surgeries,and 35 (90%) cases of level Ⅲ&Ⅳ surgeries. The index surgical procedures included 19 (49%) biliary-intestinal procedures, 11 (28%) simple intestinal procedures, and 9 (23%) anal and colorectal procedures. The direct causes of unplanned reoperation included 10 (26%) anastomotic leakages, 8 (20%) adhesive intestinal obstructions, 5 (13%) postoperative intussusceptions, 5 (13%) incisional complications (infection, dehiscence and incisional hernia), 3 (8%) postoperative hemorrhages and 8 (20%) miscellaneous (iatrogenic injury and surgical misjudgment). Patients' prognosis included 24 (62%) full recoveries, 9 (23%) further operations, and 5 (13%) deaths, and 1 (3%) short bowel syndrome.Conclusions:Most pediatric unplanned reoperations after elective digestive tract surgery occur in complex surgical procedures. The most common causes of unplanned reoperation are anastomotic leakage, adhesive intestinal obstruction. Unplanned reoperations are often prone to adverse effects on prognosis.

9.
Front Genet ; 12: 638683, 2021.
Article in English | MEDLINE | ID: mdl-34220929

ABSTRACT

Oat is an annual gramineous forage grass with the remarkable ability to survive under various stressful environments. However, understanding the effects of high altitude stresses on oats is poor. Therefore, the physiological and the transcriptomic changes were analyzed at two sites with different altitudes, low (ca. 2,080 m) or high (ca. 2,918 m), respectively. Higher levels of antioxidant enzyme activity, reactive oxygen and major reductions in photosynthesis-related markers were suggested for oats at high altitudes. Furthermore, oat yields were severely suppressed at the high altitude. RNA-seq results showed that 11,639 differentially expressed genes were detected at both the low and the high altitudes in which 5,203 up-regulated and 6,436 down-regulated. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment tests were conducted and a group of major high altitude-responsive pigment metabolism genes, photosynthesis, hormone signaling, and cutin, suberine and wax biosynthesis were excavated. Using quantitative real-time polymerase chain response, we also confirmed expression levels of 20 DEGs (qRT-PCR). In summary, our study generated genome-wide transcript profile and may be useful for understanding the molecular mechanisms of Avena sativa L. in response to high altitude stress. These new findings contribute to our deeper relevant researches on high altitude stresses and further exploring new candidategenes for adapting plateau environment oat molecular breeding.

10.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-911598

ABSTRACT

Objective:To evaluate clinical characteristics and treatment of postoperative anastomotic stricture in pediatric congenital biliary dilatation patients.Methods:The clinical data of 24 children with postoperative anastomotic stricture from Apr 2012 to Oct 2019 in Beijing Children's Hospital was retrospectively analyzed.Results:There were 6 males and 18 females. Patients were divided into bile- leak group (BL, n=6) and non bile-leak group (NBL, n=18) based on whether there was anastomotic leakage after primary surgery. The main symptoms in BL group was persistent obstructive jaundice, and recurrent cholangitis in NBL group. Postoperative symptoms were first shown in an average of 7.0 months in BL group, compared to 59.0 months in NBL group, P<0.05. In BL group, 4 underwent redoing hepaticojejunostomy, 2 underwent anastomosis plasty. In NBL group, 3 underwent redoing hepaticojejunostomy, 15 did anastomosis plasty with multiple biliary stones found necessitating extraction. After reoperation, one patient had bile leakage, 2 patients had recurrent cholangitis within one-month, 21 patients had uneventful recovery. Five were found to have biliary stones in long-term follow-up. Conclusions:Biliary-enteric anastomotic leakage can cause stricture in postoperative patients of congenital biliary dilatation ,reoperation is necessary in symptomatic patients.

11.
BMC Plant Biol ; 20(1): 565, 2020 Dec 14.
Article in English | MEDLINE | ID: mdl-33317475

ABSTRACT

BACKGROUND: Environmental lodging stress, which is a result of numerous factors, is characterized by uncertainty. However, several studies related to lodging in cereal crops have reported that lodging in the Hippeastrum rutilum environment is very rare. Hippeastrum rutilum is a garden flower with high ornamental value and abundant germplasm resources. Under past cultivation practices, it was found that the plant types of 'Red Lion', with red flowers, and 'Apple Blossom', with pink flowers, are quite different. The leaves of 'Red Lion' are upright, while the leaves of 'Apple Blossom' show lodging, which seriously affects its ornamental value. The aims of this study were to compare the differences between the two varieties with leaf lodging and upright leaves according to morphological and physiological attributes. In this study, karyotype analysis and phenotypic morphological and physiological characteristics were compared to explore the differences between the two plant types. RESULTS: The karyotype analysis of the two cultivars showed that their chromosome types were both tetraploid plants. The results showed that the lignin content in the leaves of 'Red Lion' was high, the cross-sectional structure of the leaf vascular bundle was more stable, and the chlorophyll content was high. In addition, significantly less energy was transferred to the electron transport chain (ETR) during the photoreaction. Similarly, the results regarding the maximum photosynthetic rate (Fv/Fm), nonphotochemical quenching (NPQ) and effective quantum yield of photosystem II photochemistry (△F/Fm') all indicated that the photosynthetic capacity of "Red Lion" was greater than that of "Apple Blossom", which was affected by leaf lodging. The size of the leaves was significantly smaller, and the leaf sag angle, leaf width, and leaf tip angle presented significantly lower values in 'Red Lion' than in 'Apple Blossom', which exhibits leaf sag. The difference in these factors may be the reason for the different phenotypes of the two cultivars. CONCLUSION: The results of this study proved that lodging affects the photosynthetic capacity of Hippeastrum rutilum and revealed some indexes that might be related to leaf lodging, laying a theoretical foundation for cultivating and improving new varieties.


Subject(s)
Amaryllidaceae/anatomy & histology , Amaryllidaceae/physiology , Amaryllidaceae/genetics , Plant Breeding , Plant Leaves/anatomy & histology , Plant Leaves/physiology
12.
Mycobiology ; 48(2): 104-114, 2020.
Article in English | MEDLINE | ID: mdl-32363038

ABSTRACT

The carbohydrate-active enzyme (CAZyme) genes of Trametes contribute to polysaccharide degradation. However, the comprehensive analysis of the composition of CAZymes and the biosynthetic gene clusters (BGCs) of Trametes remain unclear. Here, we conducted comparative analysis, detected the CAZyme genes, and predicted the BGCs for nine Trametes strains. Among the 82,053 homologous clusters obtained for Trametes, we identified 8518 core genes, 60,441 accessory genes, and 13,094 specific genes. A large proportion of CAZyme genes were cataloged into glycoside hydrolases, glycosyltransferases, and carbohydrate esterases. The predicted BGCs of Trametes were divided into six strategies, and the nine Trametes strains harbored 47.78 BGCs on average. Our study revealed that Trametes exhibits an open pan-genome structure. These findings provide insights into the genetic diversity and explored the synthetic biology of secondary metabolite production for Trametes.

13.
Sci Rep ; 10(1): 6415, 2020 04 14.
Article in English | MEDLINE | ID: mdl-32286483

ABSTRACT

Drought stress is the most pervasive threat to plant growth, which predominantly encumbers turf grass growth by causing alterations in plant functions. This study appraised the role of nitrogen isotopes in providing a theoretical basis for developing and improving Kentucky bluegrass cultivar performance under drought stress. Nitrogen isotopes labelled 15NH4Cl and K15NO3 were prepared to replace KNO3 in Hoagland's solution at concentrations of 15NH4+ and 15NO3 at 1.5, 15, and 30 mM; the solutions were imposed on stressed plants under glasshouse conditions. Nitrogenous nutrition reduced oxidative stress by elevating the enzymatic activities and proline contents of all three clonal ramet leaves, particularly under stress conditions. Apart from nitrogen content, nitrogen isotope abundance, relative water content and water potential within controls were enhanced in treated with 15NH4+ than in with 15NO3 in both the roots and leaves of Kentucky bluegrass. Nevertheless, an application of 15NH4Cl and K15NO3 at 30 mM had a positive influence to some extent on these attributes under drought stress. Overall, our results suggested that nitrogen isotopes contributed to drought tolerance in all three clonal ramets of Kentucky bluegrass by maintaining a better osmoprotectant and antioxidant defence system, which helped the plants eliminate reactive oxygen species.


Subject(s)
Droughts , Nitrogen Isotopes/metabolism , Poa/enzymology , Poa/physiology , Stress, Physiological , Water/metabolism , Catalase/metabolism , Peroxidase/metabolism , Plant Leaves/metabolism , Plant Roots/metabolism , Proline/metabolism , Superoxide Dismutase/metabolism
14.
International Journal of Surgery ; (12): 212-216, 2020.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-863294

ABSTRACT

Hepatic ischemia-reperfusion injury is the most common complication after liver transplantation. Oxidative stress, autophagy, and inflammatory response caused by excessive reactive oxygen species production are significant steps that cause liver ischemia-reperfusion injury. What′s more, nuclear factor erythroid 2-related factor 2 is considered to be a major regulator of the antioxidant response, the PI3K-Akt-mTOR signaling pathway is considered to be an important pathway of autophagy, and the HMGB1-TLR4-NF-κB signaling pathway is considered to be a key signaling pathway which leads to inflammation. Based on the above signaling pathways and regulatory factor, this article shows that the antioxidant, anti-inflammatory and autophagy regulation effects of genes, molecules and drugs on hepatic ischemia-reperfusion cells, to explore the protective effects on hepatic ischemia-reperfusion cells.

15.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-800474

ABSTRACT

Colorectal carcinoma (CRC) is the third most common malignancy in adults. Pediatric colorectal carcinoma (PCRC) is a rare non-embryonal tumor with a significantly lower incidence compared to adults. The clinical manifestations of PCRC are not typical, and pediatricians usually have no enough experience in diagnosis and treatment. Therefore, early diagnosis is extremely difficult, which would always lead to late clinical stages when diagnosis is made. At present, the pathogenesis of PCRC is still not clear, and many countries have started to carry out researches at the level of genes, molecules and cells. In both tumor primary tumors and distant metastases, PCRC has obvious difference in distribution from adults, and the proportion of pathological type of mucous adenocarcinoma (including the signet ring cell carcinoma) was significantly higher than that of adults. Although treated according to adult colorectal cancer guidelines, PCRC has been unable to achieve ideal efficacy with poor prognosis and lower long-term survival rate. The purpose of this paper is to summarize the epidemiological characteristics, pathogenesis, clinical symptoms, pathological types, treatment and prognosis of colorectal cancer in children by reviewing the latest literatures at home and abroad.

16.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-746248

ABSTRACT

Circular RNA (circRNA) is a ubiquitous class of noncoding RNAs (ncRNAs) in all kingdoms of life with diverse structures.CircRNA is an endogenous noncoding RNA molecule with covalently close cyclic structure which does not have 5'end cap and 3'poly (A) tail.It is found to be evolutionary conservative and stable with tissue specificity.CircRNAs have many biological functions.In addition to acting as miRNA sponges,many other functions are being revealed,including regulator of gene transcription,and splicing and protein translation.It is involved in ovarian epithelial neoplasms,pre-eclampsia and other development.CircRNAs could regulate the expression of numerous gynecological cancer-related microRNA (miRNAs).The circRNA-miRNA-messenger RNA(mRNA) axis is a known regulatory pattern of several cancer-associated pathways,with both agonist and antagonist effects on carcinogenesis.Numerous studies have shown that circRNAs may become potential targets and clinical diagnostic markers for reproductive and gynecological diseases.This review focuses on the biogenesis and properties of circRNAs,databases of circRNA,their functions and potential significance in gestational,tumor and gestational diseases.

17.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-775837

ABSTRACT

OBJECTIVE@#To establish a non-invasive method for beta-thalassemia by detecting parental CD41-42 mutation in cell-free DNA derived from maternal plasma with droplet digital PCR (ddPCR).@*METHODS@#Beta-actin gene and beta-thalassemia gene CD41-42 mutation were respectively set as the reference and target sequences. A novel method was established based on Bio-Rad ddPCR technique with specific primers and TaqMan probes for the two genes. The accuracy, sensitivity and detective linearity range of the developed method were evaluated by detection of the target gene gradient concentration samples. The applicability was also evaluated by testing 20 maternal plasma samples.@*RESULTS@#The ddPCR method could accurately detect the beta-thalassemia CD41-42 mutation in cell-free DNA derived from maternal plasma. Within the target sequence concentration ratio of 5.00%-0.50%, the relative errors were all < 0.05, the linear regression equation was Y=1.0101-X-0.0071 and R=0.9994. The results of 20 maternal plasma cell-free DNA samples were all consistent with those of the follow-up testing.@*CONCLUSION@#A ddPCR method for detecting parental CD41-42 mutation in cell-free DNA from maternal plasma was developed. The method is simple, rapid, accurate, and can be applied for non-invasive prenatal diagnosis for couples simultaneously carrying the CD41-42 mutation.


Subject(s)
Female , Humans , Pregnancy , Cell-Free Nucleic Acids , DNA , Blood , Mutation , Polymerase Chain Reaction , Prenatal Diagnosis , Methods , beta-Thalassemia , Diagnosis , Genetics
18.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-709702

ABSTRACT

Objective To evaluate the relationship between phosphatidylinositol 3?kinase∕serine?threonine kinase(PI3K∕Akt)signaling pathway and autophagy during acute lung injury in septic mice. Methods Thirty?six male C57BL∕6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups(n=12 each)using a random number table: sham operation group(group SH), sepsis group (group S)and PI3K inhibitor LY294002 plus sepsis group(group LY+S). Sepsis was induced by cecal ligation and puncture in S and LY+S groups. LY294002 30 mg∕kg was intraperitoneally injected at 2 h be?fore operation in group LY+S. Arterial blood samples were taken at 24 h after operation for blood gas analy?sis, PaO2was recorded, and oxygenation index was calculated. Lung specimens were obtained for examina?tion of pathological changes which were scored and for determination of autophagosome count(using trans?mission electron microscope), wet∕dry weight ratio(W∕D ratio)and expression of Akt, phosphorylated Akt(p?Akt), Beclin?1 and microtubule?associated protein 1 light chain 3 Ⅱ(LC3 Ⅱ). The p?Akt∕Akt ratio was calculated. Results Compared with group SH, oxygenation index was significantly decreased,and the W∕D ratio and pathological score were increased in S and LY+S groups, the autophagosome count was significantly increased, p?Akt∕Akt ratio was increased, and the expression of Beclin?1 and LC3Ⅱ was up?regulated in group S(P<0.05). Compared with group S, oxygenation index was significantly de?creased, and the W∕D ratio was increased, the autophagosome count was decreased, pathological scores were increased, p?Akt∕Akt ratio was decreased, and the expression of Beclin?1 and LC3Ⅱwas down?regu?lated in group LY+S(P<0.05). Conclusion PI3K∕Akt signaling pathway activation mediates autophagy and is involved in the endogenous protective mechanism of acute lung injury in septic mice.

19.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-708363

ABSTRACT

Objective To summarize our therapeutic experiences on patients with pediatric spontaneous biliary duct perforation.Methods We retrospectively analyzed the clinical data of patients diagnosed as spontaneous biliary duct perforation who were admitted into the Department of Pediatric General Surgery,the Beijing Children Hospital from January 2008 to December 2014,and summarized the therapeutic experiences.Results There were 7 boys and 18 girls,with a average age of 2.4 years (range 11 months to 10 years).Twenty-one patients (84.0%) were diagnosed by ultrasonography.Two patients were treated with conservative therapy and were discharged home.The remaining 23 patients were treated with emergent surgery.Of these patients,9 were treated with cholecystostomy and abdominal drainage (the cholecystostomy group),and the remaining 14 were treated with choledochal drainage and abdominal drainage (the choledochal drainage group).The mean hospitalization stay for the cholecystostomy group was 25.2 days,and 3 patients developed comphcations (33.3%).The mean hospitalization stay for thecholedochal drainage group was 16.1 day,and 2 patients developed complications (14.2%).Twenty-four patients were diagnosed to suffer from congenital choledochal cysts or pancreaticobiliary maljunction by imaging studies during or after surgery.Elective choledochal cystectomy with hepaticojejunostomy were performed on 23 stable patients who developed no severe complications.Conclusions Pediatric spontaneous bile duct perforation is closely related with congenital choledochal cysts,and the pathological basis in diagnosis is pancreaticobiliary maljunction.Bile duct elastic fiber hypogenesis and specific blood supply are important to the onset of perforation.Abdominal ultrasonography plays an important role in the diagnosis.Patients with peritoneal irritation and non-localized perforation should be operated in time,and choledochal drainage with abdominal drainage is a good treatment choice.All patients diagnosed as congenital choledochal cysts or pancreaticobiliary maljunction should undergo elective choledochal cystectomy with hepaticojejunostomy.

20.
Front Plant Sci ; 8: 983, 2017.
Article in English | MEDLINE | ID: mdl-28642781

ABSTRACT

Non-irrigated crops in temperate and irrigated crops in arid regions are exposed to an incessant series of drought stress and re-watering. Hence, quick and efficient recuperation from drought stress may be amongst the key determinants of plant drought adjustment. Efficient nitrogen (N) nutrition has the capability to assuage water stress in crops by sustaining metabolic activities even at reduced tissue water potential. This study was designed to understand the potential of proper nutrition management by studying the morphological and physiological attributes, and assimilation of nitrogen in Kentucky bluegrass under drought stress. In present study, one heterogeneous habitat and four treatments homogenous habitats each with four replications were examined during field trial. Drought stress resulted in a significant reduction in the nitrogen content of both mother and first ramets, maximum radius, above and below ground mass, number of ramets per plot, leaf water contents and water potential and increased the carbon content and the C:N ratio in both homogenous and heterogeneous plots compared to well-watered and nutritional conditions. Observation using electron microscopy showed that drought stress shrunk the vessel diameter, circumference and xylem area, but increased the sieve diameter, and phloem area in the leaf crosscutting structure of Kentucky bluegrass, first, second, and third ramet leaf. Thus, it can be concluded that water stress markedly reduced all the important traits of Kentucky bluegrass, however, proper nutritional management treatment resulted in the best compensatory performance under drought assuaging its adversity up to some extent and may be considered in formulating good feasible and cost-effective practices for the environmental circumstances related to those of this study.

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