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1.
Article in Chinese | WPRIM | ID: wpr-878857

ABSTRACT

The purpose of this study was to explore the expression pattern of miRNA in the process of embryo dormancy and provide a reference for the mechanism of regulating seed dormancy and germination by miRNA. We used high-throughput sequencing technology, bioinformatics analysis and real-time fluorescent quantitative PCR(qPCR) technology to sequence, screen and identify miRNAs of dormant and dormant embryos. The results showed that there were 23 811 977, 24 276 695, 20 611 876 and 20 601 811 unique sequences in the four sample libraries during the period of dormancy and dormancy release. MiRNAs are mainly distributed between 21 and 24 nt, among which the length of 24 nt occurred most frequently. A total of 31 known miRNAs were identified, belonging to 13 different families. 93 new miRNAs were predicted by bioinformatics software. Ten miRNAs(mir156 a-5 p, mir160 a-5 p, mir160 h-1, mir169 a-5 p, mir157 d, mir159 a-1, mir395-3, mir156 f-5 p, mir156-2 and mir171 a-3 p) were screened out. In this study, 10 miRNAs related to seed dormancy release were identified. The target genes mainly involved carbohydrate metabolism, plant hormone signal transduction, cell division and growth. The results of qRT-PCR showed that the sequencing results were consistent with the actual results.


Subject(s)
Gene Expression Regulation, Plant , Humans , Liliaceae , MicroRNAs , Plant Dormancy , RNA, Plant , Seeds
2.
Journal of Experimental Hematology ; (6): 1892-1898, 2020.
Article in Chinese | WPRIM | ID: wpr-879989

ABSTRACT

OBJECTIVE@#To study the effect of 5-aminoimidazole-4-formamide ribonucleotide (AICAR) combined with interferon (IFN-α-2b) on the proliferation and apoptosis of chronic myeloid leukemia K562 cells, and explore its possible mechanism.@*METHODS@#CCK-8 method was used to detect the inhibition of cell proliferation. Wright Giemsa method was used to stain and cell morphology was observed by light microscopy. FITC Annexin V/PI double staining method was used to analyze the change of apoptosis rate. Immunocytochemistry method was used to detect the expression of wild-type P53 protein.@*RESULTS@#Different concentration of AICAR was inhibitory effect on K562 cells at different time point of action for 24 h, 48 h, and 72 h, and the inhibition was time and dose-dependent (r=0.71, r=0.84). The combination of AICAR and IFN-α-2b could effectively inhibit the proliferation and promote apoptosis of K562 cells. The inhibition rate of K562 cells was (45.26±2.54)%, and the early apoptosis rate was (33.72±0.23)%, which was statistically significantly different from the control group, AICAR or IFN-ɑ-2b alone (P<0.05). The combination of two drugs promoted the expression of wild-type p53 protein.@*CONCLUSION@#AICAR and/or IFN-ɑ-2b can inhibit the cell proliferation and promote the apoptosis of K562 cells. The combination of two drugs shows synergistic antitumor effect, and its mechanism may be related to the promotion of high expression of wild-type p53 protein.


Subject(s)
Apoptosis , Cell Proliferation , Formamides , Humans , Imidazoles , Interferons , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Ribonucleotides/pharmacology
3.
Article in Chinese | WPRIM | ID: wpr-299349

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the difference of postoperative drainage and systemic trauma response between endoscopic and traditional near total thyroidectomy to provide the basis for selecting appropriate operative methods.</p><p><b>METHODS</b>In this prospective clinical controlled study, 80 patientsscheduled for near total bilateral thyroidectomy for the first time were divided equally into endoscopic surgery group (group A) and open surgery group (group B). The total drainage volume after operation, postoperative extubation time, and postoperative daily drainage volume were recorded after the operation. The contents of triglyceride (TG) and total protein (TP) were determined in the postoperative drainage fluid onthe first day. The levels of interleukin 6 (IL6), high sensitive C reactive protein (HSCRP), alpha 1 acid glycoprotein (AAG), ceruloplasmin (CER) and haptoglobin (HPT) in venous blood were tested before the operation and on the first day after surgery.</p><p><b>RESULTS</b>Compared with those in group B, the postoperative drainage volumein group Aincreased significantly (P=0.000) and the postoperative extubation time was significantly prolonged (P=0.000); the mean postoperative daily drainage volume was significantly larger ingroup A than in group B (P=0.000) and tended to decrease with time in both groups. There was no significant difference in the content of triglycerideortotal protein in the drainage fluid between the two groups on the first day after operation (P=0.429 and 0.324, respectively). In both groups, the contents of AAG, ceruloplasmin and haptoglobin on the first postoperative day were all similar with those measurement before operation (P>0.05), but significant variations occurred in the levels of IL6 and HSCRP on the first postoperative day (P=0.000). The serum levels of IL?6 or HS?CRP did not differ significantly between the two groups on the first day after operation (P=0.054 and 0.066, respectively).</p><p><b>CONCLUSION</b>Compared with open surgery, endoscopic near total bilateral thyroidectomyis associated with an increased the volume of postoperative drainage and a prolonged time of extubationbut not an increased systemic trauma response. Therefore, endoscopic surgery can serve as one of the routine options for patients who are concerned with neckscars resulting from open surgeries.</p>

4.
Article in Chinese | WPRIM | ID: wpr-272672

ABSTRACT

Pinellia Rhizoma is one of the most commonly used medicinal herbs in clinic, but its toxicity couldn't be ignored. Processing is a detoxification method before the toxic traditional Chinese medicine were given to the patients, and mainly impacted the amount of relevant components in Chinese medicinal herbs (increase or decrease). Although there were still some disputes about the toxic components in Pinelliae Rhizoma (mainly referring to the alkaloid substances), more literatures reported that needle-like calcium oxalate crystals and lectin protein in Pinelliae Rhizoma were the main toxic components, and had a significant effect on inflammation and irritation caused by Pinellia Rhizoma. With the development of research methods and the expansion of research angles, researches for the effect of processing on the detoxification mechanism of Pinelliae Rhizoma have been constantly deepened. The recent reports showed that the detoxification mechanism of the herb was correlated with the dosage of detoxifying components, and the effect of relevant excipients in inhibiting a variety of inflammatory cytokines. However, we shall also pay attention to alum and other processing accessories that could cause new toxicity from residual aluminum, and the impact from processing conditions, origin of Chinese medicinal herbs and their combination with other medicinal herbs on Pinellia Rhizoma's toxicity. This paper describes toxic components, different detoxification methods and relevant detoxification mechanisms in Pinelliae Rhizoma to provide the references for further research and development of the plant.

5.
Article in Chinese | WPRIM | ID: wpr-304803

ABSTRACT

In order to investigate the epigenetic variations between diploid and autotetraploid of Platycodon grandiflorus. The diploid buds of P. grandiflorus were soaked in the mixture of different concentration colchicines and 0.002 g•mL ⁻¹ dimethyl sulphoxide (DMSO).The identification of autotetraploid plants were based on morphological characteristics, chromosome number and flow cytometry. And then the level and pattern of DNA methylation explored by using the technology of methylation sensitive amplified polymorphism (MSAP).The result demonstrated that the buds soaked in 0.2% colchicines and 0.002 g•mL ⁻¹ DMSO solution for 12 h was ideal conditions to induce autotetraploid of P. grandiflorus, with induction rate of 32.0%.The diploid and tetraploid plants existed distinctly differences in morphological indexes.Totally,1 586 bands were amplified by 20 pairs of selective primers, of which 764 and 822 bands were detected in diploid and autotetraploid respectively. The total methylation ratio,full methylation ratio and hemimethylated ratio were 91.25%,61.25% and 30.65% in diploid of P. grandiflorus,respectively.However,the total methylation ratio,full methylation ratio and hemimethylated ratio of autotetraploid of P. grandiflorus were 86.13%,54.38% and 31.75%, respectively. Compared with diploid, the genomic DNA total methylate ratio and full methylation ratio of autotetration plants decreased by 6.02% and 7.14%.But the hemimethylated ratio of autotetraploid was higher than that of diploid, which more than 1.6%. All this results indicated that DNA methylation patterns have adjusted during the polyploidy process..

6.
Article in Chinese | WPRIM | ID: wpr-330264

ABSTRACT

In order to investigate the genetic basis of morphological variation of tetraploid plantlets of Atractylodes macrocephala, diploid plantlets were taken as experimental material, sterile filtration colchicine was used to soak 0.5-1.0 cm long buds. The difference between morphology and stomatal of diploid and tetraploid of A. macrocephala was compared, and genome polymorphism was explored by AFLP. The results showed that the buds dipped in 0.1% colchicine solution for 36 h was optimal conditions to induce tetraploid of A. macrocephala with induction rate of 32.0%. Morphological indexes such as leaf area index, leaf length and width, the density of stomas and the number of chloroplast of tetraploid were distinctly different from diploid. Four hundred and fifty-one bands ranging with 80-500 bp were amplified with 24 pairs of primers, the rate of polymorphism was 32.59%. These amplification sites of diploid were different from tetraploid of A. macrocephala, and the differences in morphology of them were reflected in the DNA polymorphism.


Subject(s)
Amplified Fragment Length Polymorphism Analysis , Methods , Atractylodes , Genetics , Sequence Analysis, DNA , Tetraploidy
7.
Acta Pharmaceutica Sinica ; (12): 350-354, 2011.
Article in Chinese | WPRIM | ID: wpr-348952

ABSTRACT

Rhubarb anthraquinone derivatives (AQs) have been documented to have both therapeutic and toxic effect on liver and kidney, leading to a complex puzzle to assess their benefits and risks. In this study, the tissue distributions of AQs in SD rats after orally administrated extracts of raw and prepared rhubarb were examined whether they undergo different uptake. The total rhubarb extract (14.49 g x kg(-1) of body weight per day od, counted on the quantity of crude material) was administrated orally for 12 weeks. The concentrations of the AQs in different tissues were quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The five major AQs, aloe-emodin, rhein, emodin, chrysophynol and physcion, could all be detected in the liver, kidney and spleen, while only rhein, aloe-emodin and emodin reached the quantitative limit. The tissue concentrations of AQs in raw rhubarb group were higher than that in steamed rhubarb group with rhein > emodin > aloe-emodin in the same tissue. On the whole, the tissue distribution of rhein was higher than that of emodin and aloe-emodin in liver, spleen and kidney. AQs could not be detected in those tissues after drug withdraw for 4 weeks, which suggested scarcely any accumulative toxicity of rhubarb. The result indicated that raw rhubarb had more tissue toxicity than steamed rhubarb and rhein may be one of the major poisonous ingredients. The results were concordant with the traditional Chinese medicine theory of toxicity-attenuating effect of processing.


Subject(s)
Administration, Oral , Animals , Anthraquinones , Pharmacokinetics , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Pharmacokinetics , Emodin , Pharmacokinetics , Female , Male , Plant Roots , Chemistry , Plants, Medicinal , Chemistry , Random Allocation , Rats , Rats, Sprague-Dawley , Rheum , Chemistry , Tandem Mass Spectrometry , Tissue Distribution
8.
Chinese Medical Journal ; (24): 2767-2770, 2011.
Article in English | WPRIM | ID: wpr-292808

ABSTRACT

<p><b>BACKGROUND</b>Patients with the genotypes of both CYP2C9*3/*3 and VKORC1-1639 A/A are expected to require the lowest dose of warfarin, and to have a greatly increased risk of bleeding. The experience for the dosing of warfarin in such extremely rare cases has been seldom reported.</p><p><b>METHODS</b>Demographic and clinical data from two cases with stable low dose of warfarin in China were studied by resequencing the corresponding gene segments in their whole blood DNA. The potential clinical value of the pharmacogenetic algorithm for them was evaluated by calculating the stable dose of warfarin in pharmacogenetic algorithm developed by International Warfarin Pharmacogenetics Consortium.</p><p><b>RESULTS</b>Both cases (68-year-old female and 50-year-old male) were diagnosed as chronic nonvalvular atrial fibrillation needing warfarin treatment, with target international normalized ratio (INR) 2 to 3. Case 1 had stable warfarin dose of 0.625 mg/d and case 2 1.25 mg/d. They needed more than 1 month to stabilize their anticoagulation. Exceeding INR values were recorded for them when the dose of warfarin was no more than 2 mg/d. Hemorrhagic complication appeared in case 1 when the dose was titrated from 2.5 to 1.25 mg/d. No concomitant medicine to increase or decrease the INR value was recorded for them. Genotyping CYP2C9 and VKORC1 showed both patients were the carriers of the homozygous alleles -CYP2C9*3/*3 and VKORC1-1639 A/A. Their stable doses of warfarin calculated by the pharmacogenetic dose algorithm (0.672 mg/d for case 1 and 1.16 mg/d for case 2) were comparable with their actual stable therapeutic doses.</p><p><b>CONCLUSIONS</b>Two Chinese with the rare genotypes of both CYP2C9*3/*3 and VKORC1-1639 A/A were found to require the extremely low dose of warfarin. The pharmacogenetic algorithm incorporating the variances of VKORC1 and CYP2C9 genotypes, as well as the non-genetic factors could predict their stable dose of warfarin with high accuracy.</p>


Subject(s)
Aged , Anticoagulants , Aryl Hydrocarbon Hydroxylases , Genetics , Cytochrome P-450 CYP2C9 , Female , Genotype , Hemorrhage , Humans , Male , Middle Aged , Mixed Function Oxygenases , Genetics , Pharmacogenetics , Vitamin K Epoxide Reductases , Warfarin
9.
Article in Chinese | WPRIM | ID: wpr-323624

ABSTRACT

<p><b>OBJECTIVE</b>To construct the plasmid pSG5/TRIF and investigate its expression in Huh7 cells.</p><p><b>METHODS</b>The plasmid pCX4pur/Myc-TRIF was digested with Not I and the digestion product was blunted followed by further digestion with EcoR I to obtain the insert Myc-TRIF. pSG5 was digested sequentially with Sma I and EcoR I. All the digested products were analyzed with agarose gel electrophoresis. The products with the expected size were extracted and ligated, and the positive clones were screened by ampicillin and amplified. The recombinant pSG5/TRIF was extracted, purified, and identified by restriction endonuclease BamH I and agarose gel electrophoresis. The recombinant plasmids were transfected into Huh7 cells with FuGene 6 reagents and into Huh7 cells previously infected with recombinant vaccinia virus (rVV) via Lipofectin. Immunofluorescence and Western blotting were performed to detect the expression of the recombinant plasmids, and the transfection efficiency with different transfection reagents was compared.</p><p><b>RESULTS</b>BamH I digestion resulted in a fragment with the expected size. Immunofluorescence staining showed successful expression of Myc-TRIF protein in Huh7 cells, and the transfection efficiency was enhanced in Huh7 cells previously infected with rVV. SDS-PAGE analysis showed that the relative molecular mass of the expressed product by pSG5/Myc-TRIF was about 100 ku, and prior infection of the cells with rVV obviously increased transfection efficiency, as was consistent with the results of immunofluorescence.</p><p><b>CONCLUSION</b>pSG5/Myc-TRIF is successfully constructed and expressed in Huh7 cells. The expression efficiency can be increased by prior infection of the cells with rVV.</p>


Subject(s)
Adaptor Proteins, Vesicular Transport , Genetics , Cell Line, Tumor , Gene Expression , Genetic Vectors , Hepacivirus , Genetics , Humans , Plasmids , Recombinant Fusion Proteins , Genetics , Transfection
10.
Chinese Journal of Pediatrics ; (12): 126-129, 2007.
Article in Chinese | WPRIM | ID: wpr-349477

ABSTRACT

<p><b>OBJECTIVE</b>To investigate whether Helicobacter pylori infection has any effects on the epithelial cell proliferation, apoptosis and P53 gene expression as well as its role in the pathogenesis of chronic gastritis.</p><p><b>METHODS</b>Sixty children with chronic gastritis were studied. All the children underwent upper digestive tract endoscopy and biopsy specimens were taken. Helicobacter pylori infection was determined with microscopic examination after Gimsa staining and the rapid urease test and 30 of the children were Helicobacter pylori positive and the other 30 were negative. The relation between the findings and cell proliferation was studied by immunostaining; the status of gastric apoptosis was tested by DNA fragmentation in situ using TdT-mediated dUTP biotin nick end labeling (TUNEL). Immunohistochemical method was used to detect the expression of P53 protein; CagA antibody was tested by Western blotting.</p><p><b>RESULTS</b>(1) The proliferative index and apoptosis index in children with Helicobacter pylori infection with CagA positive gastritis were much higher than those of Helicobacter pylori negative gastritis patients [(11.56 +/- 4.21)% vs. (5.85 +/- 2.21)%, (10.58 +/- 5.31)% vs. (2.86 +/- 0.64)%, P < 0.01]. (2) The proliferative index and apoptosis index in 30 cases with Helicobacter pylori infection with CagA positive gastribis were much higher than 21 cases who were cured by effective drugs [(11.50 +/- 4.11)% vs. (3.74 +/- 2.30)%; (10.58 +/- 4.02)% vs. (3.74 +/- 2.30)%, P < 0.01]. (3) The expression of P53 protein in Helicobacter pylori with CagA positive gastritis children was much higher than that of Helicobacter pylori negative cases [(63% vs 16%), P < 0.1].</p><p><b>CONCLUSION</b>CagA positive Helicobacter pylori infection with gastritis improved gastric epithelial cell proliferation and apoptosis. The abnormal expression of P53 protein in gastric epithelium may play an important role in regulation of the processes.</p>


Subject(s)
Antibodies, Bacterial , Blood , Antigens, Bacterial , Allergy and Immunology , Apoptosis , Bacterial Proteins , Allergy and Immunology , Biopsy , Cell Proliferation , Child , Child, Preschool , Epithelial Cells , Metabolism , Female , Gastric Mucosa , Pathology , Gastritis , Pathology , Helicobacter Infections , Pathology , Helicobacter pylori , Humans , In Situ Nick-End Labeling , Male , Tumor Suppressor Protein p53 , Metabolism
11.
Article in Chinese | WPRIM | ID: wpr-638984

ABSTRACT

Objective To investigate the changes of blood levels of gastrin(GAS),motilin(MTL) and insulinoid growth factor-1(IGF-1) in critical children with multiple organ disfunction syndrome(MODS).Methods The levels of GAS,MTL and IGF-1 in 50 critically ill children and 30 children without critical illness were detected.The control group consisted of 30 health children.All serum or plasma levels of GAS,MTL and IGF-1 were measured by radioimmunoassay.Results The serum or plasma levels of GAS,MTL and IGF-1 were significantly higher in critically ill children than those in uncritically ill children and control group(F=49.61,55.18,18.23 all (P

12.
Chinese Journal of Hepatology ; (12): 654-656, 2003.
Article in Chinese | WPRIM | ID: wpr-339130

ABSTRACT

<p><b>OBJECTIVE</b>To study the relationship between aldehyde dehydrogenase (ALDH) gene polymorphism and alcoholic liver disease, and investigate the genetic pathogenesis of alcoholic liver disease (ALD).</p><p><b>METHODS</b>PCR, restriction endonuclease and electrophoresis were used, to detect the genotypes and alleles frequencies of ALDH gene in patients in the control group, alcohol dependent group and ALD group, and each group contained 20 patients.</p><p><b>RESULTS</b>The frequencies of ALDH2*1 and ALDH2*2 allele had statistic significance between control group and ALD group (x2=4.80, P<0.05), and no statistic significance between control group and alcohol dependent group. ALDH2*1/*1 was predominant in alcohol dependent group and ALD group, while ALDH2*2/*2 was not detected.</p><p><b>CONCLUSIONS</b>The gene polymorphism of ALDH is close to ALD. The allele of ALDH2*2 may be a negative risk factor for the developing of ALD</p>


Subject(s)
Adult , Aldehyde Dehydrogenase , Genetics , Alleles , Gene Frequency , Humans , Liver Diseases, Alcoholic , Genetics , Male , Polymorphism, Genetic , Risk Factors
13.
Article in Chinese | WPRIM | ID: wpr-638478

ABSTRACT

Objective To investigate the early-pathologic changes in children′s antrum infected with different types of Hp and study the Hp isolate′s pathogenic.Methods The serum types of CagA and VacA from Hp were determined by Westen-Blot in 70 patients with Hp positive and 36 patients with Hp negative.The standard of gastritis pathologic classification was accordance with that of international made in Sydney. The pathogenic of Hp affecting was evaluated by the degree of inflammation, severity of active gastritis,lymph follicles and atrophy.Results The detection rate of type Ⅰwith high virulence in Hp isolates was 68.1%,mid-type isolates was 27.7% and type Ⅱ with low virulence isolates was 4.2%.To observe the pathologic distinction in 49 patients with type Ⅰisolate,20 patients with mid-type isolate and 3 patients with type Ⅱ isolate,the type Ⅰ and mid-type isolates had significant difference in inflammation and their activity in either antrum or duodenal ampulla.Three patients with type Ⅱ isolate have not active gastritis.Type Ⅰand mid-type isolates had significant difference in lymph follicles,and the lymph follicles caused by type Ⅰwere significant higher than those caused by mid-type.But there were no significant differences in intestinal metaplasia and atrophy.Conclusions TypeⅠisolate with high virulence is the main detection isolate of children infected by Hp in our district.There is inflammation occurrence in antrum specimens in childhood who infected with Hp.

14.
Article in Chinese | WPRIM | ID: wpr-638490

ABSTRACT

Objective To study the life quality of 2 - 3 years old survivors in Neonatal Intensive Care Unit (NICU). Methods Severe neonates were randomly assigned to intervention group (group 1,30 cases) and non- intervention group (group 2,30 cases) depending on the early intervention applied or not,as well as 30 healthy newborns as normal controls. The physical,neurological conditions and intelligence test were taken regularly. To investigate the psychological state, actions, temperament and family conditions when they were2-3 years old.Results Mental development index(MDI) and physical development index(PDI) in early interventional group were significant higher than those in group 2(P

15.
Article in Chinese | WPRIM | ID: wpr-638325

ABSTRACT

Objective To study the clinical significance and changes of plasma nitric oxide (NO) and cyclic adernosine monophosphate(cAMP)/cyclic guanosine monophosphate(cGMP) ratio in children with asthma.Methods NO,cAMP,cGMP levels and changes of cAMP/cGMP in 40 asthmatic children in acute attack stage and restoration stage and in 23 health children were measured by nitric acid reductase method and radioimmunoassay.Results 1.the amount of NO 3-/NO 2-in the plasma of asthmatic children in acute attack stage was obviously higher than that in restoration stage and in normal control group(P0.05);6.the results of linear correlation analysis showed that there were positive correlations between plasma NO 3-/NO 2- and plasma cGMP levels in acute attack stage (r=0.401 P

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