The effect of preoperative use of intra-aortic balloon pump in acute myocardial infarction without cardiogenic shock patients undergoing off-pump coronary artery bypass grafting / 中华胸心血管外科杂志

Objective To evaluate the efficacy and safety of preoperative intra-aortic balloon pump(IABP) insertion in acute myocardial infarction(AMI) without cardiogenic shock(CS) patients receiving off-pump coronary artery bypass grafting ( OPCABG).Methods 444 consecutive AMI patients who underwent isolated OPCABG from January 2009 to December 2016 were enrolled.158 patients who underwent preoperative IABP placement(IABP group) and the other of 286 patients who did not have IABP placement(control group).The in-hospital mortality rate, postoperative complications, mechanical ventilation time, ICU stay and hospital length were compared between the two groups.Results The overall mortality was 5.0%.135 pairs of patients were matched.The preoperative IABP insertion showed benefits in postoperative survival rate compared with the control group(0 vs.5.9%, P＝0.004).However, patients with preoperative IABP were more likely to prolong duration of mechanical ventilation and ICU stay.The postoperative length of stay in hospital didn't show significant difference between the two groups.Conclusion Survival advantage was observed from preoperative IABP insertion in AMI patients without CS under-going OPCABG.

Effects of DAPT on biological activity of hematopoietic stem cells in mice / 中国实验血液学杂志

This study was to investigate the effects of DAPT (N-[N-(3,5-difluorophenacetyl-L:-alanyl)]-S-phenylglycine-butyl ester) on cell cycle, apoptosis, differentiation and expansion of hematopoietic stem cells (HSC) of mouse and to elucidate the possible mechanisms. The mRNA expressions of cell cycle-related genes p18, p21, p27, CDK1, CDK2, CDK4, CDK6, and apoptosis-related genes Bcl-2, Bcl-xl, mcl-1, Bax, Bim, p53, Puma were measured by real-time PCR. The cell cycle and apoptosis of Lin(-)c-kit(+)Sca-1(+) marked cells and CD34(-)Lin(-)c-kit(+)Sca-1(+) marked cells in bone marrow cells were detected by flow cytometry. The differentiation level of HSC was determined by single cell culture. The expansion of HSC were measured with long-term culture. The results indicated that the mRNA expression of the cell cycle related-genes CDK1, CDK2, CDK4, CDK6, p27 in Lin(-) c-kit(+)Sca-1(+) marked cells increased (P < 0.05), the expression of p18, p21 decreased (P < 0.05), the expression of the apoptosis related-genes Bcl-2, Bcl-xl, Bax, p53, Puma in Lin(-) c-kit(+)Sca-1(+) marked cells increased (P < 0.05), the expression of Bim decreased (P < 0.05), the expression of Mcl-1 had not changed (P > 0.05) after treatment with DAPT 1 µmol/L for 5 d. The changes of cell cycle of Lin(-)c-kit(+)Sca-1(+) marked cells in bone marrow had no statistical significance after treatment with DAPT 1 µmol/L for 5 d, CD34(-)Lin(-)c-kit(+)Sca-1(+) marked cells in bone marrow at G(0) phase decreased and at G(1) phase increased after treatment with DAPT 1 µmol/L for 5 d (P < 0.05); the apoptotic fractions of Lin(-) c-kit(+) Sca-1(+) marked cells and CD34(-)Lin(-)c-kit(+)Sca-1(+) marked cells in bone marrow increased after treatment with DAPT 1 µmol/L for 5 d (P < 0.05). The changes of colony number, average number of cells in wells and their differentiation had no statistical significance (P > 0.05) after treatment with DAPT 1 µmol/L for 10 d. Expansion of HSC in bone marrow of mouse decreased after treatment with DAPT 1 µmol/L for 3 d. It is concluded that DAPT not only enhances the exhaustion of CD34(-)Lin(-)c-kit(+)Sca-1(+) marked cells in bone marrow cells of mouse, but also enhances the apoptosis of Lin(-)c-kit(+)Sca-1(+) marked cells and CD34(-)Lin(-)c-kit(+)Sca-1(+) marked cells in bone marrow cells of mouse. DAPT also reduces the expansion of HSC. However, the changes of survival and differentiation of single CD34(-)Lin(-)c-kit(+)Sca-1(+) marked cells in mouse bone marrow cells have no statistical significance.

Differential expression of oxidative reductases in different subsets of mouse hematopoietic cells / 中国实验血液学杂志

Hematopoietic stem cells (HSC) are the source of all blood cells, which can differentiate into various hematopoietic hierarchy cells. Physiological level of reactive oxygen species (ROS) plays an important role in regulating functions of HSC as excessive ROS is harmful to HSC. Oxidative reductases and antioxidants can eliminate cellular ROS to maintain ROS homeostasis and thus avoid excessive ROS-caused damages. There are several types of oxidative reductases in cells such as catalase, manganese superoxide dismutase (MnSOD), glutathione peroxidase 1 (GPX1), thioredoxin reductase 1 (Txrnd1) and Nqo1 [NAD(P)H dehydrogenase quinone 1]. However, the functional roles of various oxidative reductases in regulating ROS level in hematopoietic cells remain unclear. This study was to investigate the expression patterns of these oxidative reductases in mouse hematopoietic cells that were sorted out via flow cytometry and to find out important oxidative reductases involving in HSC ROS regulation. The expression of various oxidative reductases was detected by semi-quantitative real-time PCR. The results showed that the expression level of catalase in T cell population was 0.14 times that in LT-HSC population (P < 0.05). The expression levels of MnSOD in CLP population and myeloid cells were 0.56 and 0.47 times that in LT-HSC population respectively (P < 0.05). The expression levels of GPX1 in ST-HSC, GMP, Myeloid cells, MEP, T lymphocytes and B lymphocytes were 1.79, 2.96, 2.07, 0.58, 0.10, 0.6 times that in LT-HSC population respectively (P < 0.05). The expression levels of Txrnd1 in ST-HSC, MPP, CMP, GMP, Myeloid cells, T lymphocytes and B lymphocytes were 3.36, 3.18, 4.19, 6.39, 4.27, 0.016, 0.56 time that in LT-HSC population, respectively (P < 0.05). The expression levels of Nqo1 in ST-HSC, MPP, CMP, GMP, CLP and B cell were 0.30, 0.17, 0.25, 0.10, 0.04, 0.01 times that in LT-HSC population, respectively (P < 0.05). It is concluded that the expression levels of oxidative reductases (catalase, MnSOD, GPX1, Txrnd1 and Nqo1) in hematopoietic hierarchy cells are cell-type specific. It suggests that reductases may play divergent roles in various hematopoietic cell populations. More importantly, the expression level of Nqo1 in LT-HSC population significantly increased as compared with other cell populations, thereby suggesting its unique regulatory role in HSC.

Observation of CT three-dimensional reconstruction in the treatment of standard manipulation for lumbar intervertebral disc herniation / 中国骨伤

<p><b>OBJECTIVE</b>To observe the morphological change of prominence through CT three-dimensional reconstruction before and after manipulative treatment and in order to investigate biomechanical effect of manipulation in treating lumbar intervertebral disc herniation (LIDH).</p><p><b>METHODS</b>From December 2009 to May 2010, 24 patients with LIDH (32 herniated discs) with the unilateral typing,which were treated with manipulation (on alternate day one time and every time about 20 min, 3 weeks as a course of treatment). There were 10 males and 14 females, ranging in age from 25 to 54 years with an average of 36.2 years, in course of disease from 2 days to 10 years with an average of 6.9 years. Protrusible 12 discs were in L4,5 and 20 discs were in L5S1. According to typing of distance between prominence and zygapophysial joint or vertebral plate (ligamentum flavum), 5 cases were type I, 13 cases were type II and 6 cases were type III. After a course of treatment,the morphological changes of prominences were analyzed in the same level of CT three-dimensional reconstruction, including contour map of nerve root sheath side distance (TD), the distance between prominence and zygapophysial joint or vertebral plate (ligamentum flavum), the deviated angle of prominence (AN value) and the sagittal index (SI value).</p><p><b>RESULTS</b>From the contour map of TD, 19 patients (79.2% of the total) can be identified morphological changes after the treatment; from the distance between prominence and zygapophysial joint or vertebral plate (ligamentum flavum), 7 cases with type II turned into type I and 2 cases with type III turned into type II after treatment; AN value increased after treatment (P<0.05),it showed prominence occurred morphological change toward deviated direction of intervertebral foramina; there was no significant difference in SI value between before and after treatment (P>0.05).</p><p><b>CONCLUSION</b>Standard manipulation can make prominence change, the prominence and nerve roots release, and mutual position improve,which can provide imaging evidence for the study in biomechanical effects.</p>

Construction, expression and immunogenicity analysis of a Tat N-terminus-deleted mutant fusion protein of human immunodeficiency virus type 1 / 病毒学报

In the study, a gene encoding Tat protein N terminal 1- 21 amino acid residues-deleted mutant (Tat22-101) was amplified by PCR from a full length Tat gene of human immunodeficiency virus type 1, and the prokaryotic expression plasmid pET32a-Tat22-101 was constructed. After identification by digestion with endonucleases and sequencing, the recombinant plasmid pET32a-Tat22-101 was transformed into E. coli BL21(DE3) and expressed with IPTG induction. The mutant fusion protein with deleted Tat N terminal was purified by an affinity chromatography column Ni(2+)-NTA and subsequently identified by SDS-PAGE and Western blotting. The results showed that the molecular weight of the mutant protein was approximately 26.9kD. Furthermore, BALB/c mice were immunized with the mutant protein and the anti-sera were collected. ELISA results showed that the mutant protein preserved its immunogenicity, particularly it could improve the production of antibodies to other epitopes in addition to the N terminal epitope of Tat protein, which might provide some valuable information for the study of Tat functions as well as for development of potential novel HIV Tat vaccine.

Expression of ADAR1 isoforms in murine acute T-ALL leukemia model / 中国实验血液学杂志

This study was purposed to investigate the expression of ADAR1 isoforms of P110 and P150 during the development of murine leukemia. A Notch1 over-expressing murine T cell acute lymphoblastic leukemia model was used to study the expression of ADAR1. BMMNC were isolated at different stages of disease and CD45.2(+)GFP(+) leukemia cells were sorted by flow cytometry at late stage. The expression of ADAR1 was detected by real time quantitative PCR. The results showed that mouse bone marrow cells from both leukemia and control groups expressed P110 and P150. Difference of P110 and P150 mRNA expression were observed during the development of leukemia. The expression of P110 dramatically increased and was significantly higher than that in control group. However, the expression level of P150 in leukemia group decreased stably and reached one-fourth of that in control group at 14 day. Furthermore, similar expression patterns could be detected in sorted CD45.2(+)GFP(+) leukemia cells. It is concluded that the mRNA expressions of P110 and P150 show diverse patterns in the development of leukemia, suggesting that RNA editing mediated by ADAR1 isoforms may play different roles in leukemia.

Altered CD28 and CD95 mRNA expression in peripheral blood mononuclear cells from elderly patients with primary non-small cell lung cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The expression of the co-stimulatory molecule CD28 and death receptor CD95 on T cells, which change with age, are considered as important immunological parameters of immunosenescence. It is well established that CD28 and CD95 are associated with tumorgenesis and tumor progression, but the relationship between the age-related changes of these two immunological markers and cancer in the elderly is largely unknown.</p><p><b>METHODS</b>The levels of CD28 and CD95 mRNA in peripheral blood mononuclear cells (PBMCs) from sixty-three elderly patients (aged > or = 60 years) with primary non-small cell lung cancer (NSCLC) were analyzed by real-time fluorescence-based quantitative polymerase chain reaction (FQ-PCR). In addition, twenty young patients (aged < 60 years) with NSCLC, thirty elderly healthy donors and thirty young healthy donors were enrolled as controls.</p><p><b>RESULTS</b>CD28 mRNA levels were significantly lower and CD95 mRNA levels were significantly higher in elderly patients with NSCLC than in the other groups. Similar results were found in elderly healthy donors comparing with young healthy donors. By Logistic regression analysis an increased risk of NSCLC was markedly associated with aging, down-regulation of CD28 mRNA and up-regulation of CD95 mRNA, and CD28 mRNA had an obvious negative correlation with the CD95 mRNA. In addition, the mRNA levels of CD28 and CD95 in the peripheral blood of the elderly patients was closely associated with the tumor node metastasis (TNM) stages, grade of cell differentiation and lymph node metastasis status, but not related to pathological types.</p><p><b>CONCLUSIONS</b>The results suggest a close relationship between T cell senescence and NSCLC tumour progress in the elderly, and that up-regulation of CD28 mRNA or down-regulation of CD95 mRNA in peripheral blood T cells may play an important role in inhibiting oncogenesis and development of primary NSCLC in the elderly.</p>

Precautions of physical performance requirements and test methods during product standard drafting process of medical devices / 中国医疗器械杂志

The major idea of this article is to discuss standardization and normalization for the product standard of medical devices. Analyze the problem related to the physical performance requirements and test methods during product standard drafting process and make corresponding suggestions.

The influence of Ara-C on anti-CD3/anti-Pgp mediating T-lymphocytes activities against multi-drug resistant leukemia cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the role of Ara-C in regulating anti-CD3/anti-Pgp mediating T-lymphocytes activities against multi-drug resistant leukemia cells.</p><p><b>METHODS</b>The diabody of anti-CD3/anti-Pgp was purified by E-tag affinity chromatography. K562 and K562/A02 cells were treated with Ara-C. The expressions of B7-1 and B7-2 on K562 and K562/AO2 cells were detected by FACS. The cytotoxicity of T-lymphocytes combined with anti-CD3/anU-Pgp plus Ara-C was analyzed by CytoTox 96 nonradioactive method.</p><p><b>RESULTS</b>The expressions of B7-1 and B7-2 on K562 and K562/A02 cells treated by Ara-C was significantly higher than those untreated. The effect/target ratio was from 0.39:1 to 25:1, and the killing rate of activated T cells to anti-drug-resistant leukemia cells was from (16.44 +/- 1.20)% to (60.49 +/- 2.90)%. The killing rates were increased gradually, with both the effect/target ratio and the antibody concentration increasing (P < 0.05).</p><p><b>CONCLUSION</b>Ara-C may be an important adjuvant for improving anti-CD3/anti-Pgp mediating T-lymphocytes activities against multi-drug resistant leukemia cells.</p>

Preparation and activity detection of monoclonal antibody against anti-CD3 ScFv / 中国医学科学院学报

<p><b>OBJECTIVE</b>To prepare monoclonal antibody (McAb) against anti-CD3 ScFv for purifying and detecting serum anti-CD3 antibody concentration.</p><p><b>METHODS</b>McAb against anti-CD3 ScFv was prepared by hybridoma technique and used to prepare affinity chromatography column, which was used to purify anti-CD3 ScFv and Diabody [CD3 x Pgp] without E-tag. The binding activities of anti-CD3 ScFv, Diabody [CD3 x Pgp] without E-tag, and Diabody [CD3 x Pgp] purified by anti-CD3 affinity chromatography column or anti-E-tag affinity chromatography column against K562/A02 cell and Jurket cells were detected by fluorescence activated cell sorting (FACS) method. ELISA was used to identify the specificity of the McAb.</p><p><b>RESULTS</b>McAb against anti-CD3 ScFv specifically detected serum anti-CD3 ScFv without reacting with sera. The anti-CD3 ScFv purified by anti-CD3 affinity chromatography column and purified by anti-E-tag affinity chromatography column had the same specific binding activity with Jurkat cells. The positive binding rates of Diabody [CD3 x Pgp] without E-tag to K562/A02 and Jurkat cells were 89.87% and 83.95%, respectively. In the competitive binding experiments with K562/A02 and Jurkat cells, the binding rates of Diabody [CD3 x Pgp] without E-tag decreased to 56.30% and 43.78%, respectively.</p><p><b>CONCLUSION</b>The McAb against anti-CD3 ScFv prepared in our lab can be used to purify and detect serum anti-CD3 antibody concentration.</p>

Three-step purification of preparative-scale antiCD20 (Fab')2 / 中国医学科学院学报

<p><b>OBJECTIVE</b>To establish a three-step purification method of preparative-scale antiCD20 (Fab')2 using AKTA prime.</p><p><b>METHODS</b>AntiCD20 (Fab')2 was extracted by hyperosmotic solution and then purified by CM sepharose FF, phenyl sepharose FF, and protein G sepharose FF.</p><p><b>RESULTS</b>Around 8 mg anti-CD20 (Fab')2, whose purification was 96.678%, was purified. The antigen-binding activity of antiCD20 (Fab')2 was similar to that of antiCD20 (Fab')2 purified by protein G sepharose FF and S-100.</p><p><b>CONCLUSION</b>The three-step purification method can obtain high-purity preparative-scale antiCD20 (Fab')2 in a simple way.</p>

Effect of catalpol from Radix Rehmanniae on A?_(25-35)-induced apoptosis of PC12 cells / 上海第二医科大学学报

Objective To investigate the effect of catalpol from Radix Rehmanniae on A?25-35-induced apoptosis of PC12 cells.Methods PC12 cells were routinely cultivated and treated by A?25-35(final concentration,20 ?mol/L) 24 hours after the addition of catalpol or saline.Forty-eight hours later,cells were examined for viability and apoptosis by MTT method and TUNEL method,respectively,while Bax and Bcl-2 mRNA expression were analyzed by semi-quantitive RT-PCR. Results Catalpol could significantly elevate the viability at 1?10-5 mol/L and 1?10-4 mol/L(P

Cloning and expression of the extracellular domain of 4-1BBL / 生物工程学报

RT-PCR was used to clone DNA fragment of the extracellular domain of 4-1BBL from human THP-1 cells (human monocyte), and the expression vector pAYZ4-1BBL was constructed by cloning the extracellular domain of 4-1BBL into the expression vector pAYZ. The extracellular domain of 4-1BBL was expressed in E. coli 16C9 and purified by affinity chromatography. SDS-PAGE and Western blot analysis showed that the relativae molecular weight of soluble 4-1BBL is 22kD which was consistent with the theoretically predicted value. So far as we know, it is the first time that the soluble expression of 4-1BBL in E. coli was achieved 4-1BBL induced a significant release of IL-2 in stimulated Jurkat cells after 48h incubation, especially in the presence of tumor cell. At the same time the apoptosis level of Jurkat cell reduce more than 50%. In conclusion, 4-1BBL may be useful in cancer immunotherapy.

High level expression of chimeric antibody fragment F(ab')2 directed against CD20 in Escherichia coli / 生物工程学报

The use of tumor antigen specific antibody for the delivery of therapeutic agents offers the possibility of targeting therapy with reduced toxicity to normal tissues compared to conventional treatments. In previous work, the human-mouse chimeric antibody fragment Fab' directed against CD20 was constructed from the new anti-CD20 antibody HI47 (a mouse IgG3, K). The chimeric antibody fragment Fab' could reduce its antigenicity, but the yield, quality and affinity of chimeric antibody fragment Fab' restrict its use. To improve affinity of chimeric antibody fragment Fab', a new phasmid pYZcpp3, which expresses chimeric antibody fragment F(ab')2, was constructed by adding a sequence encoding a small peptide, (CPP)3, to C-terminus of heavy chain constant region of chimeric antibody fragment Fab'. Using the pYZcpp3 to transform E. coli. 16c9, the genetically engineered bacteria 10916# was obtained. 10916# can secret the soluble chimeric antibody fragment Fab' and F(ab')2 into periplasmic. The yield was up to 360 mg/L with the percent of F(ab')2 up to 45% in 19L fermentor by the high density fermentation technology. Without denaturation and renaturation, the F(ab')2 has possessed the native three-dimensional structure. The purity of F(ab')2 was more than 90% after the purification of protein G affinity chromatography and S200 size exclusion chromatography. The F(ab')2 could distinguish and bind to Raji cells (CD20+) by FACS. F(ab')2 could inhibit the proliferation of Raji cells in vitro by MTT, IC50 was 22.8 microg/mL. HI47 and its chimeric fragments F(ab')2 induced a significant level of apoptosis (23.5%, 20.8%, respectively), independent of any cross-linking agents, in Raji cells after 24 h incubation. The chimeric antibody fragment F(ab')2 directed against CD20 is possible to apply to tumor therapy in clinic in the future.

Study on effect of arsenic trioxide on adhesion and invasion of human hepatocarcinoma cells in vitro / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore that the arsenic trioxide injection (ATI) has the effect in antagonizing adhesion and invasion of human hepatocarcinoma cells (HCC), and its relevant mechanism.</p><p><b>METHODS</b>Human hepatocellular carcinoma cell line SMMC-7721 and the high metastatic nude mice human HCC in situ transplantation model was taken as the objects of study, the effects of ATI on the SMMC-7721 cell movement and migration, its adhesion with fibronectin (FN) and endothelial cell (EC), as well as the CD44 and MMP-2 gene protein expression in transplanted tumor of the model mice were observed by means of cell movement and migration test, cell adhesion test and immunohistochemical method.</p><p><b>RESULTS</b>ATI could significantly inhibit SMMC-7721 cell movement and migration on FN, adhesion with FN and EC, also could lower CD44 and MMP-2 in cancer cells.</p><p><b>CONCLUSION</b>ATI has the effects of antagonizing hepatocarcinoma cell adhesion and invasion, the mechanism may be related with the action of ATI in lowering CD44 and MMP-2 expression in cancer cells.</p>

Effects of 3-substituted aryl oxindole(PH II-7) on cell cycle of tumor cells / 药学学报

<p><b>AIM</b>To study the antitumor mechanism of 3-substituted aryl oxindole (PH II-7) and determine its effects on cell cycle distribution of tumor cells.</p><p><b>METHODS</b>The cell cycle distributions were determined with FACS. The cell cycle regulation-related proteins of K562 lysates were analyzed with Western Blot. The inhibition of PH II-7 on DNA synthesis of tumor cells were estimated though 3H-thymidine incorporation and the tyrosine kinase activity of EGFR of A431 lysates was measured with ELISA.</p><p><b>RESULTS</b>PH II-7 effected cell cycle distribution of several tumor cells, including multidrug resistant tumor cell lines, and accumulation of cells in the G0-G1 stages was observed. The cell cycle regulation-related proteins CDK2, Rb and c-myc were inhibited by PH II-7 in a dose dependent manner, whereas the expression of CyclinE was increased after exposure to PH II-7. Furthermore, PH II-7 2.0 mg.L-1 was shown to inhibit the incorporation of 3H-thymidine into DNA, and 21.89%-41.29% of the PTK activity of EGFR in A431 lysates was inhibited by PH II-7 2-8 mg.L-1 in a dose-dependant manner.</p><p><b>CONCLUSION</b>PH II-7, a new anti-tumor agent, blocks the transition of cell cycle of tumor cells from G1 to S phase by inhibition CDK2.</p>

CLONING OF FULL CRY1C GENE FROM B.T.C.AND EXPRESSION IN CROPS BENEFICIAL BACTERIA BACILLUS CEREUS / 微生物学通报

Specific primers 1CaA/1CaB for full cry1C gene in Bacillus thuringiensis subsp.colmeri strain 15A3 were designed. The 4.0kb PCR product included the whole ORF and regulation region of cry1C gene. This PCR product was linked with shuttle vector pHT315 by two cloning steps. The recombined plasmid pHT-1C was electroporated into Bacillus cereus 9509, a kind of bacteria that beneficial to crops. The transformant could produce bipyramidal-shaped parasporal inclusions. The 60kD protein band was detected by SDS-PAGE. The bioassay result showed that the cry1C gene transformant of Bc 9509 had insecticidal activity to Spodoptera exigua.