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Objective: To clone CDS sequence of Dioscorea opposita SUS gene and analyze the protein structure,in order to provide a theoretical basis for the regulation mechanism of SUS gene and the synthesis mechanism of D.opposita polysaccharides.Method: Total RNA in D.opposita was extracted and reverse-transcribed into first strand of cDNA.Specific primers were designed according to an annotated SUS gene sequence obtained from the laboratory D.opposita genome database,and the coding region of the SUS gene was obtained by gene cloning technique and the protein sequence characteristics were analyzed by protein prediction analysis software.Result: A 2 448 bp gene sequence was cloned with a complete open reading frame (ORF).The gene was named DoSUS1.The formula of protein encoded by DoSUS1 gene in D.opposita was C4209H6534N1115O1205S23,and the molecular weight was 9 788.32,the total number of amino acids was 815,the theory isoelectric point (PI) was 6.10,the extinction coefficient was 110 505,the aliphatic index (AI) was 94.15,the instability index was 32.18,and the grand average of hydropathicity (GRAVY) was-0.225.It was a stable and soluble acidic protein.There were phosphorylation sites in the DoSUS1 amino acid sequence,with no transmembrane region and signal peptide.The secondary structure and the tertiary structure showed that DoSUS1 was an α class protein.Functional domain predictions showed that DoSUS1 had sucrose synthesis domain and glycosyl transfer domain.The homology alignment showed that the amino acid sequence of DoSUS1 was more than 80% similar to the amino acid sequence of the aligned monocots.The phylogenetic tree showed that DoSUS1 was closely related to SUS of monocotyledon evolution.Conclusion: The coding sequence of SUS gene was cloned from D.opposita for the first time,and its protein structure was analyzed to lay a foundation for further studying the roles of SUS in the growth and polysaccharide synthesis of D.opposita.
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Objective To study the expression of CD269 and CD317 antigens in bone marrow cells of patients with multiple myeloma (MM),analyze its correlation with the laboratory indexes reflecting the progression of MM and evaluate its value in clinical diagnosis and treatment.Methods 63 newly diagnosed MM patients were selected as the study group by a casecontrol study.The expression rate of CD269 and CD317 in bone marrow blood of 35 patients with iron deficiency anemia and other antigens in bone marrow blood of 63 patients with MM were detected by flow cytometry.The levels of serum hemoglo bin (Hb),serumβ2-MG(β2-MG) and lactatedehydrogenase (LDH) in patients with MM were dctectcd,and the levels of CD269 and CD317 were analyzed statistically.Results The positive rates of CD269 in the study group and control group were (86.6±2.35)％ vs (4.33±l.69)％,rcspectivcly (t =4.256,P＜0.05)).The positive rate of CD317 was (71.42+ 0.62)％ vs (8.32+ 3.89)％,the difference was statistically significant (t=3.102,P＜0.05).In other expression,the expression level of CD269 and CD317 in CD56 positive group was significantly higher than that of negative group (t=4.032,P＜0.05),while the expression of CD117 the level of positive group was significantly lower than that of the negative group (t 2.832,P＜0.05),CD19,CD20 expression was not statistically significant difference between the two groups (P＞ 0.05).The levels of CD269 and CD317 in patients with MM were positively correlated with the level of CD56 expression (r =0.392,P＜0.05),and negatively correlated with the level of CD117 expression (r=-0.210,P＜0.05).The levels of CD269 and CD317 in patients with MM were significantly lower than those in the negative group (t=3.012,P＜0.05) and the levels of serum LDH in the positive group were lower than those in the negative group (t=2.024,P＜0.05).There was a negative correlation between Hb content (r=-0.212,P＜0.05) and negatively correlated with serum β2-MG (r=-0.312,P＜0.05).Conclusion The high expression of CD269 and CD317 in bone marrow cells in MM patients is related to the increase of CD56 and decrease of CD117 in patients with MM.
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<p><b>OBJECTIVE</b>To investigate the effect of decitabine and plasma of ITP patients on in vitro cultrue of megakaryocytes in bone marrow of steroid-resistant ITP patients.</p><p><b>METHODS</b>Bone marrow mononuclear cells were isolated from 20 steroid-resistant ITP patients, both methyl cellulose semisolid culture system (to observe and count the number of megakaryocytes colony-forming unit) and liquid culture system (to analysis the expression rate of CD41a(+) cells) were used for megakaryocyte cultrue. The experiments were divided into 4 groups according to the different components of the culture system, group A was control, group B was added with decitabine, group C with ITP plasma, group D with both decitabine and ITP plasma, and the rest of the culture components were the same in the 4 groups except the above-mentioned materials. Morphology of megakaryocytes was observed by inverted and light microscopy. The expression rate of CD41a⁺ cells in culture was analysed by flow cytometric.</p><p><b>RESULTS</b>Different concentration of decitabine showed different effect on megakaryocyte growth of steroid-resistant ITP patients and the optimal concentration to differentiate into megakaryocyte for bone marrow mononuclear cells is 3.0 µmol/L. Compared with group A, both megakaryocyte colony forming units (CFU) and expression rate of CD41a⁺ cells in group B were statistically significantly higher (P < 0.05). As compared with group A, the megakaryocyte colony-forming units in group C decreased with statistically significant difference, while compared with group C, the megakaryocyte colony-forming units in group D obviously increased with statistically significant difference.</p><p><b>CONCLUSIONS</b>Decitabine is able to induce bone marrow mononuclear cells of steroid-resistant ITP patients to differentiate into megakaryocyte and the optimal concentration is 3.0 µmol/L; ITP plasma is able to inhibit the megakaryocyte growth of steroid-resistant ITP patients.</p>
Subject(s)
Humans , Azacitidine , Bone Marrow , Bone Marrow Cells , Drug Resistance , Flow Cytometry , Hematopoietic Stem Cells , Inosine Triphosphate , Megakaryocytes , Stem Cells , SteroidsABSTRACT
Heuristic teaching is propitious to cultivating students' ideation. We have used different heuristic modes for example problems, stories, contrast, associationin and so on to cultivate medical students' ideation in pathobiology and immunology teaching and acquired good effect.
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The digital music editor software "Cool Edit Pro 2.0" is used to design a virtual hearing testing system. This system has following advantages. First, its signal frequency can be set at will. Second, its dynamic range of signal intensity can reach up to 80dB. Third, the measuring accuracy of decibel value may reach 0.1dB. Forth, the system can be used in single and dual channel measurements. Last but not least, it can carry on data processing and drawing along with the same computer.
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Audiometry , Computers , Hearing Tests , Music , Signal Processing, Computer-Assisted , Software , User-Computer InterfaceABSTRACT
<p><b>OBJECTIVE</b>To explore the influence of lanthanum chloride (LaCl3) on inducible nitric oxide synthase (iNOS) expression in RAW264.7 macrophages with lipopolysaccharide (LPS) induction, and to investigate its possible mechanisms.</p><p><b>METHODS</b>The RAW264.7 macrophages were randomly divided into four groups: i. e, control group (without treatment), LaCl3 group (with treatment of 2.5 micromol/L of LaCl3 for 24 hrs), LaCl3 + LPS group (with treatment of 2.5 micromol/L LaCl3 for 24h), and LPS group (with treatment of 1 mg/L LPS for 24 hrs). The iNOS protein expression was measured by immunofluorescence and Western blot. iNOS gene expression was assayed by reverse transcription-polymerase chain reaction (RT-PCR). NO production in culture supernatant was assayed by nitrate reductase method.</p><p><b>RESULTS</b>Immunofluorescence analysis showed that iNOS was located mainly in the cytoplasm. RAW264.7 cells with overexpression of iNOS accounted for 44.4%, which was obviously higher than that in LaCl3 + LPS group (11.8%, P < 0.05). There was a faint signal of FITC-labeled green tint in control group or LaCl3 group. The iNOS mRNA and protein expression, and the NO content in LPS group were significantly higher than those in control, LaCl3, and LaCl3 + LPS groups (P < 0.05).</p><p><b>CONCLUSION</b>LaCl3 can suppress LPS-induced iNOS overexpression at mRNA and protein level and reduce NO production, indicating that LaCl3 can antagonize the excessive activation of iNOS induced by LPS.</p>