Association between blood pressure during 12-28 weeks gestation and pre-eclampsia: predictive value of blood pressure trajectories constructed by latent class growth modeling / 中华心血管病杂志

Objective: To explore the associations between blood pressure trajectories during pregnancy and risk of future pre-eclampsia in a large cohort enrolling pregnant women at gestational age of ~12 weeks from community hospitals in Tianjin. Latent class growth modeling (LCGM) was used to model the blood pressure trajectories. Methods: This was a large prospective cohort study. The study enrolled pregnant women of ~12 weeks of gestation in 19 community hospitals in Tianjin from November 1, 2016 to May 30, 2018. We obtained related information during 5 antepartum examinations before gestational week 28, i.e., week 12, week 16, week 20, week 24 and week 28. LCGM was used to model longitudinal systolic (SBP) and diastolic blood pressure (DBP) trajectories. For the association study, the predictors were set as SBP and DBP trajectory membership (built separately), the outcome was defined as the occurrence of preeclampsia after 28 weeks of gestation. Results: A total of 5 809 cases with known pregnant outcomes were documented. After excluding 249 cases per exclusion criteria, 5 560 cases with singleton pregnancy were included for final analysis. There were 128 cases preeclampsia and 106 cases gestational hypertension in this cohort. Univariate logistic regression and multivariate logistic regression showed the higher baseline SBP level and DBP level were related with increased risk of preeclampsia. Four distinctive SBP trajectories and DBP trajectories from 12 weeks to 28 weeks of gestation were identified by LCGM. After controlling for potential confounders (baseline BMI, being primipara or not, white blood cell counts, hemoglobin level, platelet counts and alanine aminotransferase level), the OR for SBP latent classification trajectory_ 4 was 4.023 (95%CI: 2.368 to 6.835, P<0.001), and the OR for SBP latent classification trajectory_3 was 1.854 (95%CI: 1.223 to 2.811, P=0.004). Logistic regression showed that: using the DBP latent classification trajectory_1 as the reference group, the OR for DBP latent classification trajectory_4 was 4.100 (95%CI: 2.571 to 6.538, P<0.001), and 2.632 (95%CI: 1.570 to 4.414, P<0.001) for DBP latent classification trajectory_2. After controlling for potential confounders (baseline BMI, being primipara or not, white blood cell counts, hemoglobin level, platelet counts and alanine aminotransferase level), the OR for DBP_traj_4 was 2.527 (95%CI: 1.534 to 4.162, P<0.001), and the OR for DBP_traj_3 was 1.297 (95%CI: 0.790 to 2.128, P=0.303), and 2.238 (95%CI: 1.328 to 3.772, P=0.002) for DBP_traj_2. Therefore, BP trajectories from 12 weeks to 28 weeks identified by LCGM served as novel risk factors that independently associated with the occurrence of preeclampsia. Receiver operating characteristic (ROC) curve analysis showed incremental diagnostic performance by combing baseline blood pressure levels with blood pressure trajectories. Conclusion: By applying LCGM, we for the first time identified distinctive BP trajectories from gestational week 12 to 28, which can independently predict the development of preeclampsia after 28 weeks of gestation.

PD-1 expression, mRNA level and cytotoxicity changes in CD19CAR-T cells / 中华血液学杂志

Objective: To observe the changes of PD-1 expression, mRNA level and cytotoxic activity of CD19 CAR-T cells during the culture process of CAR-T cells. Methods: The peripheral blood T cells of 6 lymphoma patients with high expression of PD-1 and 6 healthy volunteers were the source of CAR-T cells. The expression of PD-1 was analyzed by flow cytometry. The mRNA level of PD-1 was analyzed by PCR. The cell proliferation was analyzed by CCK-8 assay. The cytotoxicity was analyzed by LDH assay. Results: ①The transfection efficiency of high PD-1 expression T cells and healthy volunteer T cells were as the same (P>0.05) . ②The cell proliferation capacity of CD19 CAR-T cells from high PD-1 expression T cells or healthy volunteer T cells, with or without PD-1 inhibitor were as the same (P>0.05) . ③The cytotoxicity to lymphoma cells of high PD-1 expression T cells and CAR-T cells were lower than that of these two T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer T cells (P<0.001) . There was no difference of the cytotoxicity between the CAR-T cells from high PD-1 expression T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer (P>0.05) . ④There was no difference of the expression of PD-1 in all CAR-T cell groups during the culture process (P>0.05) . There was no difference of mRNA level of PD-1 in all groups during the culture process (P>0.05) . ⑤The PD-1 expression of CAR-T cells increased by the time of culture after contacting with lymphoma cells (P<0.001) . The PD-1 inhibitors could antagonize this effect. There was no difference of mRNA level of PD-1 in all groups after contacting with lymphoma cells (P>0.05) . Conclusion: The PD-1 expression of CAR-T cells from high PD-1 expression T cells increased by the time of culture after contacting with lymphoma cells. However, the mRNA level of PD-1 of all groups did not change, even if PD-1 inhibitor was applied.

Efficacy and safety of CD19 chimeric antigen receptor T cells for the treatment of 22 patients with B-cell lymphoma / 中华血液学杂志

Objective: To investigate the efficacy and safety of CD19 chimeric antigen receptor T (CAR-T) lymphocytes for the treatment of B cell lymphoma. Methods: A total of 22 patients with B-cell lymphoma from February 1, 2017 to July 1, 2018 were reviewed to evaluate the efficacy and adverse reactions of CD19 CAR-T. Results: Of 22 patients with B-cell lymphoma received CD19 CAR-T cells, the median dose of CAR-T cells was 7.2 (2.0-12.0) ×106/kg. Nine of 12 cases of relapse refractory patients were overall response. Complete remission (CR) occurred in 2 of 12 patients, partial remission (PR) in 7 of 12 patients. The overall response in minor residual disease positive (MRD) group was 8 of 10 patients. CD19 CAR-T cells proliferated in vivo and were detectable in the blood of patients. The peak timepoints of CAR-T cells proliferated in the relapsed refractory and MRD positive groups were 12 (5-19) and 4.5 (1-12) days after treatment respectively, and among peripheral blood cells, CAR-T cells accounted for 10.10% (3.55%-24.74%) and 4.02% (2.23%-28.60%) of T lymphocytes respectively. The MRD positive patients achieved sustained remissions during a median follow-up of 8 months (rang 3-18 months) . None of all the patients relapsed during a median follow-up time of 10 months (3-18 months) . However, 7 PR responders of the relapsed refractory patients maintained a good condition for 1.5-6.0 months. One patient bridged to hematopoietic stem cell transplantation, another one sustained remission for 12 months. Cytokine-release syndrome (CRS) occurred in 14 patients with grade 1-2 CRS in MRD positive group and grade 3 CRS in relapsed refractory group. Conclusions: CAR-T cell therapy not only played a role in the rescue treatment of relapsed and refractory patients, but also produced a surprising effect in the consolidation and maintenance of B-cell lymphoma. CD19 CAR-T cells might be more effective in the treatment of MRD positive B-cell lymphoma patients than in the refractory or relapsed cases. High response rate was observed with fewer adverse reactions.

Diagnosis and treatment of hypertensive disorders in pregnancy according to changes from guidelines / 中国实用内科杂志

Hypertensive disorders in pregnancy(HDP)is a group of diseases including high blood pressure during pregnancy. Strengthening clinical monitoring, risk factors assessment and early intervention would promote outcomes of women and their children. Blood pressure and proteinuria are key monitoring during pregnancy. Antihypertensive drugs recommended for pregnancy include methyldopa, labellol, or nifedipine. Early screening and intervention of preeclampsia is one of the keys to the prevention of cardiovascular risk during pregnancy. In addition to medication, weight control, screening and treatment of obstructive sleep apnea(OSA), lifestyle intervention and so on can not be ignored.

Effect of PD-1 inhibitor Nivolumab on the proliferation and cytotoxicity of anti-CD19 chimeric antigen receptor T cells / 中华血液学杂志

Objective: To Evaluation the effect of PD-1 inhibitor Nivolumab on the proliferation and cytotoxicity of anti-CD19 chimeric antigen receptor T cells (CD19-CAR-T) in vitro. Methods: Five patients with high PD-1 expression in peripheral blood and five healthy volunteers were selected. These peripheral blood mononuclear cells were used as the source of T cells to prepare CD19-CAR-T cells. Different doses (72, 36, 18 μg/ml) of Nivolumab was added on day 8 to the culture medium. Patient T cells incubated with 72 μg/ml Nivolumab and CD19-CAR-T cells of healthy volunteers were used as controls. CCK-8, lactate dehydrogenase (LDH) cytotoxicity assay and ELASA were used to detect the proliferation capacity, the specific cytotoxicity and the inflammatory factor secretion. Results: ①T cells from patients with high expression of PD-1 as the source of CD19-CAR-T cells did not affect transfection rate compared with that of healthy volunteers [(32.80±7.22)% vs (35.10±5.84)%, t=-0.554, P=0.593]. ②Incubation of CD19-CAR-T cells with 72 μg/ml Nivolumab did not affect CD19-CAR-T cell proliferation, but its cytotoxicity was significantly higher than that of CD19-CAR-T cells alone or patients' T cells +72 μg/ml Nivolumab (all P<0.001), there was no significant difference in the killing activity between the 72 μg/ml and 36 μg/ml Nivolumab treated CD19-CAR-T cells on Pfeiffer cells (P=0.281, 0.267, respectively), and they were all higher than those of 18 μg/ml Nivolumab treated CD19-CAR-T cells (all P<0.001). ③Different doses of PD-1 inhibitor Nivolumab combined with CD19-CAR-T cells does not affect the secretion of IFN-γ and IFN-α (all P>0.05). Conclusion: Combination of 36 μg/ml PD-1 inhibitor and CD19-CAR-T cells could reduce the drug toxicity and enhance the cytotoxicity.

Hepatocytes apoptosis induced by β1-adrenoceptor autoantibody is involved in hepatic dysfunction / 基础医学与临床

Objective To investigate the effect of β1-Adrenoceptor autoantibody on liver function.Methods The biologically active of β1-AA was prepared and passive immunization model was established with β1-AA.The bio-chemical parameters of the liver were measured by the automatic serum biochemical analyzer.The liver size, hepatic vein,portal vein velocity were detected by liver ultrasound;hepatocytes apoptosis were tested by tunel stai-ning,annexin V/PI staining and caspase 3 activity detection.Results The biologically active of β1-AA and passive immunization model were established successfully.The ALT and AST of the liver significantly increased and the ALB decreased in the passive immunization process.The apoptosis of the hepatocytes increased,and meto-prolol partially reversed this effect.Conclusions β1-AA may induce hepatocytes apoptosis by β1-adrenergic receptor and participate in the development of liver injury.

Research advances in the regulation of cardiovascular metabolic disorders and its related risk factors by C1q/TNF related proteins / 生理学报

The complement C1q/TNF related protein (CTRP) family is rapidly growing and currently comprises 15 members. Although CTRP proteins share a common structure composed of four distinct domains: a signal peptide at the N terminus, a short variable region, a collagenous domain, and a C-terminal globular domain, which is homologous to adiponectin, each CTRP has a unique tissue expression profile and varied function. In this review we focus on the biochemistry and pleiotropic functions of CTRPs as new molecular mediators regulating cardiovascular metabolic disorders and its related risk factors diseases.

Expression and roles of the epithelial mesenchymal transition markers E-cadherin and vimentin in hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To determine the differential protein expressions of epithelial mesenchymal transition (EMT) markers E-cadherin and vimentin in hepatocellular carcinorma (HCC) and to investigate their correlation to the molecular mechanisms of metastasis to explore their potential utility as prognostic indicators of HCC.</p><p><b>METHODS</b>Tumor tissues and patient-matched adjacent non-tumor tissues were collected from individuals diagnosed with HCC. E-cadherin and vimentin protein expressions in the tissue specimens were quantified by western blot with densitometry of fluorescence emission and comparatively analyzed to determine the associations with molecular and clinical features. The expressions of E-cadherin and vimentin, as well as the other EMT-related protein Twist, were also detected in the tissue specimens by immunohistochemistry. Statistical analyses were carried out by paired-samples t-test, Mann-Whitney test, and Spearman rank correlation analysis.</p><p><b>RESULTS</b>E-cadherin expression was significantly lower in tumor tissues (0.082 +/- 0.063 vs. adjacent non-tumor tissues: 0.226 +/- 0.215, t = -4.050, P less than 0.01), lower in patients with portal vein tumor thrombus (vs. non-thrombic HCC patients, P = 0.001), and correlated with TNM stage (III/IV > I/II, P = 0.003). Vimentin expression was significantly higher in tumor tissues (vs. adjacent non-tumor tissues, P = 0.002), negatively correlated with E-cadherin expression (t = -0.509, P = 0.004), and closely associated with some clinical parameters, such as portal vein tumor thrombus (P less than 0.01), TNM stage (P less than 0.01), and Milan criteria (P = 0.005). Immunohistochemistry showed that E-cadherin expression was very weak in tumors but very strong in the cell membranes of non-tumor tissues, and that vimentin and Twist expressions were strong in tumors but undetectable in non-tumor tissue.</p><p><b>CONCLUSION</b>Expression levels of the EMT markers E-cadherin and vimentin in HCC are related to clinical parameters, including portal vein tumor thrombus and TNM stage, and may represent useful prognostic markers of metastasis.</p>

Effect and mechanism of iron-catalyzed oxidative stress on mesenchymal stem cells / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore effect of iron overload on the proliferation and apoptosis of mesenchymal stem cell(MSCs) and the possible mechanism.</p><p><b>METHODS</b>Iron overload model of MSCs was established by adding ferric ammonium citrae (FAC) into the culture medium at different concentrations (100, 200, 400 Μmol/L) and incubated for different lengths of time (12, 24, 48 h). The levels of labile iron pool (LIP) and reactive oxygen species (ROS) were measured to confirm oxidative stress state in the model. Changes in cell proliferation and apoptosis after iron overload were measured through population double time(DT)and annexin V-PI assay. Finally, the expressions of phosphorylated p38 mitogen activated protein kinase (P-p38MAPK), p38MAPK, protein kinase B (AKT), and p53 were determined through Western blot analysis to investigate which ROS-mediated signaling pathway was involved in this process.</p><p><b>RESULTS</b>The LIP level of MSCs was significantly increased by FAC treatment at 400 Μmol/L (mean fluorescence intensity 482.49±20.96 vs. 303.88±23.37, P<0.05). The level of intracellular ROS was positively correlated with the concentration of FAC and reached a peak level when cultured with 400 Μmol/L FAC (P<0.05).After treatment with 400 Μmol/L FAC at different time points (12 h, 24 h, and 48 h), the DT of MSCs was (1.47± 0.11) d, (1.80±0.13) d, and (2.04±0.14) d, respectively, which was signifcantly longer than that of the control, which was(1.20±0.05)d (P<0.05).The apoptosis rate was also significantly higher in iron overload group[(3.51±1.17)% vs.(0.66±0.62)%, P<0.05]with consequent increase in the expressions of P-p38MAPK, p38MAPK, and p53 proteins in iron overload group, while no significant difference was found in the expression of AKT.</p><p><b>CONCLUSION</b>Iron overload can inhibit the proliferation of MSCs and induce their apoptosis through the generation of ROS, which is probably due to the stimulation of p38MAPK- p53 signaling pathway.</p>

Dynamic expression of toll like receptor 2 and 4 in a rat model of myocardial ischemia/reperfusion injury / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the role of toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4) in myocardial ischemia/reperfusion injury (MI/RI) by observing the dynamic expression changes at mRNA and protein levels early after myocardial ischemia/reperfusion (I/ R).</p><p><b>METHODS</b>The Wistar rats were randomly divided into Sham and I/R group (n = 42), and killed according to different reperfusion time (1, 2, 4, 6, 12, 24 h and 7 d). Structural and morphous changes of myocytes were observed under optical microscope. The mRNA and protein levels of TLR2 and TLR4 were detected using real-time PCR (RT-PCR). Monocyte chemokine protein-1 (MCP-1) and interleukine-6 (IL-6) mRNA levels were measured by reverse transcriptase-polymerase chain reaction (rt-PCR).</p><p><b>RESULTS</b>(1) With the extension of reperfusion time, the myocardial infarct size increased smoothly, and reached the plateau at 4 h, then stayed in the platform. After reperfusion for 7 d, the ventricular had been remodeled. (2) At the beginning of reperfusion, myocardial structure showed no significant change in Sham group, but had different degrees of injury in I/R group. In rats of the group reperfused for 7 d the left ventricular remodeling could be visible. (3) Compared to sham group,TIR2, TLR4, MCP-1, IL-6 mRNA level were increased in myocardium in I/R group. TLR2 and TLR4 both peaked at 4 h of reperfusion, IL6 peaked at 6 h, followed by a gradually decrease. TLR4 and IL-6 mRNA levels rose again at 7 d. MCP-1 level in I/R group remained fairly with sham group at the beginning of reperfusion, and markedly elevated at 7 d.</p><p><b>CONCLUSION</b>Expression of TLRs mRNA in myocardium during early after myocardial ischemia/reperfusion increased rapidly and activated TLRs might play an important role in MI/RI through promoting the generation of inflammatory factors. At the late reperfusion, TLRs levels raise again and the expression of inflammatory factors increase once again, Those may probably affect the remodeling of ventricular, and injure myocardial structure and function.</p>

Epidemiology and antimicrobial resistance of clinical isolates about hospital infection from patients with hematological diseases / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the epidemiology and antibiotic resistance of isolates from hospitalized patients with hematological disease from 2005 to 2011.</p><p><b>METHODS</b>A total of 1453 bacterial strains were isolated from patients with hematological disease from January 2005 to December 2011. Antimicrobial susceptibility testing was performed by micro-dilution method.</p><p><b>RESULTS</b>(1) The majority of the bacterial strains were respiratory passage examples (57.5%). The portage of blood examples in our division (13.60%) was higher than of whole hospital (6.26%), with lower positive rate of bacterial culture (52.37%) than of whole hospital (60.24%). Chemotherapy-induced agranulocytosis was the main reason for hospital infection. 578 (39.8%) bacterial strains were gram positive, and 875 (60.2%) gram negative bacillus. Staphylococcus epidermidis strains and glucose nonfermenters had a tendency of ascensus. (2) Methicillin resistant staphylococcus aureus (MRSA) accounted for 72.8% antibiotic resistance. Detection rates of ESBLs in Escherichia coli and Klebsiella pneumoniae were 18.9% and 10.4%, respectively. (3) No obvious changes of antimicrobial resistances of Staphylococcus and Enterococcus were observed during these years. The Enterobacteriaceae strains showed lowest resistance rates to Carbapenems, next to Cefoperazone/sulbactam and Piperacillin/tazobactam. But the resistance rates of Escherichia coli to Cefepime and Ceftazidime were gradually increasing during the past years. Pseudomonas aeruginosa and Acinetobacter baumannii of glucose nonfermenters showed lowest resistance rates to Cefoperazone/sulbactam, but the resistance rate of Pseudomonas aeruginosa to Carbapenems increased.</p><p><b>CONCLUSIONS</b>Escherichia coli was the highest in quantity of gram negative bacillus and glucose nonfermenters had a tendency of ascensus. The resistance rates of Escherichia coli to Cefepime and Ceftazidime, Pseudomonas aeruginosa to Carbapenems were gradually increasing in the past years.</p>

Effects of humanized interleukin 21 on anti-leukemic activity of cytokine induced killer cells and the mechanism / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the effects of humanized interleukin 21 (IL-21) on anti-leukemic activity of cytokine induced killer(CIK) cells derived from peripheral blood(PB) and the mechanism.</p><p><b>METHODS</b>Mononuclear cells were separated from peripheral blood and cultured with cytokines to induce CIK cells. Proliferation of CIK cells with or without IL-21 stimulation and their cytotoxic activity against K562 cells was measured by MTT method. IL-21 receptor (IL-21R) and immunophenotypes of CIK cells were measured by flow cytometry. The expression of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), tumor necrosis factor-β (TNF-β), perforin, granzyme A, granzyme B, FasL and NKG2D mRNA were measured by semi-quantitative RT-PCR. FasL on the surface of CIK cells and intra-cellular perforin and granzyme B of CIK cells were measured by flow cytometry. The concentration of IFN-γ and TNF-α in the cultured supernatant were measured by enzyme immunoassay. JAK-STAT signalling pathway of CIK cells were measured by Western-blot.</p><p><b>RESULTS</b>After IL-21 stimulation, the proportion of CIK cells increased from (17.5 ± 4.7)% to (26.5 ± 2.1)%. Cytotoxic activity against K562 cells by CIK cells increased from (22.8 ± 2.8)% to(44.6 ± 8.3)%. The expression of IL-21R increased about 2 folds. The mRNA expression of IFN-γ increased almost 2 folds from (0.3760 ± 0.2358) to (0.7786 ± 0.2493), TNF-α increased almost 2 folds from (0.6557 ± 0.1598) to (1.3145 ± 0.2136), perforin increased almost 1.5 folds from (0.6361 ± 0.1457) to (0.9831 ± 0.1265), granzyme B increased almost 2 folds from (0.4084 ± 0.1589) to (0.7319 ± 0.1639), FasL increased almost 2 folds from (0.4015 ± 0.2842) to (0.7381 ± 0.2568), the expression of granzyme A, TNF-β and NKG2D were similar with control. Flow cytometry analysis showed that the expression of FasL of CIK cells was higher than that of control (0.19% vs 0.04%), the expression of perforin increased from 35.28% to 53.16%, and the expression of granzyme B increased from 43.16% to 78.82%. The concentration of IFN-γ in the culture supernatant increased almost 2 folds from (25.8 ± 6.1) ng/L to (56.0 ± 2.3) ng/L, and TNF-α increased almost 3 folds from (5.64 ± 0.61) µg/L to (15.14 ± 0.93) µg/L. Western blot showed that the expression of STAT1 and STAT5a had no significant differences, but the expression of STAT3 and STAT5b were higher than that of control.</p><p><b>CONCLUSION</b>Humanized IL-21 could enhance the anti-leukemic activity of CIK cells via increasing IL-21R, perforin, granzyme B, FasL, IFN-γ and TNF-α, as well as activating JAK-STAT signaling pathway. These data indicate that IL-21 has a potential clinical value in the enhancement of anti-leukemic immunotherapy.</p>

Reversion of multidrug resistance by CIK in K562/ADR cells and its mechanism exploration / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects and mechanism of cytokine-induced killer (CIK) cells in reversing multidrug resistance (MDR) and increasing intracellular concentration of adriamycin (ADR) in the K562/ADR cells.</p><p><b>METHODS</b>Peripheral mononuclear cells (MNCs) were isolated from healthy donors and cultured with combined cytokines to generate CIK. The changes of cell phenotype and cytokines secretion of CIK were determined. K562/ADR cells were divided into three groups: ADR in combination CIK (group I), CIK alone (group II) and ADR alone (group III). The viability and proliferation of K562/ADR cells were assayed by MTT assay, the intracellular concentration of ADR and the expression of P-glycoproteins (P-gp) in K562/ADR cells by FCM.</p><p><b>RESULTS</b>The cytotoxicity of ADR in group I was higher than that in group II (P < 0.05). The cytotoxicity was increased with the E/T ratio increasing (P < 0.05) but had no relation with the concentration of ADR in group I (P > 0.05). The expression of P-gp was declined in group I and group II (P > 0.05). The intracellular concentration of ADR in group I was higher than that in group II (P < 0.05), and had no relation with the ADR concentration (P > 0.05).</p><p><b>CONCLUSION</b>Pre-treatment with CIK can increase the cytotoxicity and the intracellular concentration of ADR and decrease the expression of P-gp in K562/ADR cells in the ADR and CIK combination group. Acute leukemia patients would be most likely to benefit from the combination of chemotherapy and CIK therapy.</p>

Correlation between MPO 129 A/G polymorphism and severity of coronary artery disease / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the relationship between myeloperoxidase (MPO) 129 A/G promoter polymorphisms and the severity of coronary artery disease (CAD).</p><p><b>METHODS</b>The study enrolled 267 patients who had been diagnosed as coronary artery diseases by coronary angiography. The serum MPO activity was detected by colorimetric method. PCR-RFLP method was used to decide the genotypes of the patients. The severity of CAD was evaluated by the numbers of stenotic coronary arteries and the Gensini scores respectively.</p><p><b>RESULTS</b>The MPO 129 locus G and A alleles frequency were 0.893 and 0.107, respectively. No significant difference was observed in serum MPO activity between different genotypes (P > 0.05). The distribution of genotypes in different Gensini score groups had no significant difference (P > 0.05). The patients with GG genotypes were prone to develop mutivessel diseases.</p><p><b>CONCLUSION</b>No significant correlation exists in MPO 129 locus polymorphism and serum MPO activity. The MPO 129 locus polymorphism isn't a reasonable predict factor of CAD severity.</p>

In vitro effect of iron overload on bone marrow cell function by inducing the reactive oxygen species / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the in vitro effect of iron overload on the generation of reactive oxygen species (ROS) and of bone marrow (BM) cell function.</p><p><b>METHODS</b>BM mononuclear cells (BMMNCs) were cultured with ferric citrate (FAC) at different concentrations and for different time to create iron overload and confirmed by the detection of cellular labile iron pool (LIP). The changes of ROS, apoptosis, hematopoietic colony formation (CFU-E, BFU-E, CFU-GM and CFU-mix) and the percentage of the CD34 + cells percentage were analyzed. The differences of these index were tested after the iron overload treated with deferasirox (DFO) or antioxidants (N-acetyl-L-cysteine, NAC).</p><p><b>RESULTS</b>1) When BMMNCs were cultured with FAC, the LIP was found to increase in a time and concentration dependent manner. The intracellular LIP reached maximum at 400 micromol/L of FAC for 24 hours. 2) The ROS of total cells, leukocytes and erythrocytes increased to 1.77, 1.75 and 2.12 fold respectively compared with that of normal control when cells were cultured at 400 micromol/L of FAC for 24 hours . DFO and NAC could reduce the ROS efficiently (P<0.05). 3) The apoptotic rates of the FAC treated cells [(24.80 +/- 2.99)%] increased significantly compared with that of normal control [(8.90 +/- 0.96)%]. The capacity of hematopoietic colony formation in FAC treated cells decreased markedly compared with that of normal control (P<0.05). The percentage of CD34+ cells of FAC treated cells [(0.39 +/- 0.07)%] also decreased significantly compared with that of normal control [(0.91 +/- 0.12)%]. And these changes could be recovered by addition of NAC or DFO.</p><p><b>CONCLUSION</b>Iron overload can affect the hematopoiesis by inducing the generation of ROS and this damage could be corrected by removing the excess iron and ROS of the BM cells. These findings might improve the treatment of dyshematopoiesis in patients with iron overload.</p>

Establishment of iron overloaded bone marrow model in vitro and its impact on hematopoiesis / 中国实验血液学杂志

This study was to establish an iron overload bone marrow (BM) model by co-culturing the mononuclear cells from BM with iron, and investigate its hematopoiesis changes. The iron overload model was set up by adding different concentration of ferric citrate (FAC) into the mononuclear cells from BM and culturing for different time, and the model was confirmed by detecting labile iron pool (LIP). Then the apoptosis of hematopoietic cells, ability of hematopoietic colony forming (CFU-E, BFU-E, CFU-GM and CFU-mix) and percentage of the CD34(+) cells of the BM cells all were determined. The changes of these indexes were tested after the iron-overloaded BM was treated with deferasirox (DFO). The results showed that after BM cells were cultured with FAC at different concentrations for different time, the LIP increased in time-and concentration-dependent manners. The intracellular LIP reached maximum level when cultured at 400 µmol/L of FAC for 24 hours. The detection of BM cell hematopoietic function found that the apoptotic rate of the FAC-treated cells (24.8 ± 2.99%) increased significantly, as compared with normal control (8.9 ± 0.96%)(p < 0.01). The ability of hematopoietic colony forming in FAC-treated cells decreased markedly, as compared with normal control (p < 0.05). The percentage of CD34(+) cells of FAC-treated cells (0.39 ± 0.07%) also decreased significantly, as compared with normal control (0.91 ± 0.12%)(p < 0.01). And these changes could be alleviated by adding DFO. It is concluded that the iron-overloaded model has been set by adding iron into the mononuclear cells from BM in vitro, and the hematopoietic function of iron-overloaded BM is deficient. These changes can be alleviated by removing the excess iron from the BM cells through treating with DFO. These findings would be helpful to further study the mechanism of iron-overload on the hematopoiesis of BM and also useful to find the way to treat iron-overload patients with hematopoietic disorders.

Expressions of cardiac sympathetic norepinephrine transporter and beta1-adrenergic receptor decreased in aged rats / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Evidence suggests that the deterioration of communication between the sympathetic nervous system and cardiovascular system always accompanies the aging of human and animals. Cardiac sympathetic norepinephrine (NE) transporter (NET) on presynaptic membrane is a predominant component to eliminate released NE in the synaptic cleft and maintains the sensitivity of the beta-adrenergic receptor (beta-AR). In the present study, we investigated NET and beta1-AR mRNA levels and sympathetic nerve density in cardiac sympathetic ganglion and left ventricular myocardium in 2- and 16-month-old rats with Northern blot analysis and immunohistochemistry. The expression levels of NET mRNA, NET protein and beta1-AR mRNA in the ganglia or myocardia of 16-month-old rats were markedly reduced by 67%, 26%, and 43%, respectively, in comparison with those in 2-month-old rats. Our results also show that aging induces a strong decrease of the catecholaminergic nerve fiber density.

Effects of interleukin 21 on anti-leukemia activity of cytotoxic T lymphocytes induced by dendritic cells / 中国实验血液学杂志

This study was aimed to explore the effects of interleukin 21 (IL-21) on the anti-leukemia activity of cytotoxic T lymphocytes (CTL) induced by dendritic cells (DCs) in vitro. The peripheral mononuclear cells from leukemia patients in complete remission were cultured with the specific cytokines to induce the production of DCs. The DCs loaded with RNA from autologous leukemic cells as antigen, and co-cultured with autologous T lymphocytes to get leukemia specific CTL. The cytotoxic activity of CTL against autologous leukemic cells was measured by LDH release method. The concentration of IFN-gamma and TNF-alpha in the culture supernatant was measured by enzyme immunoassay. The effects of IL-21 on the mature DCs were also studied by the measurement of the phenotype of DC and the allogenic mixed lymphocytic reactions induced by DCs. Experiments were divided into 2 groups: test group in which IL-21 (200 ng/ml) was added in coculture of DC/CTL and control group in which no IL-21 (200 ng/ml) was added. The results showed that when cultured with IL-21, the quantity of CTL increased from (56.73 +/- 10.21)% (control group) to (73.43 +/- 18.01)% (p < 0.01); The concentration of IFN-gamma and TNF-alpha in the culture supernatant increased from (154.91 +/- 67.20) ng/L (control group) to (310.62 +/- 141.15) ng/L (p < 0.01) and from (8.77 +/- 5.09) microg/L (control group) to (15.25 +/- 6.56) microg/L (p < 0.01) respectively. At the effector: target ratio of 20:1, the cytotoxic activity against autologous leukemic cells by CTL increased from (50.22 +/- 5.07)% (control group) to (75.38 +/- 9.47)% (p < 0.01). IL-21 had neither effect on the phenotype (CD1a, CD83, CD86, CD80 and HLA-DR) of mature DCs nor the allogeneic mixed lymphocytic reactions induced by DCs. It is concluded that IL-21 can strengthen the proliferation of CTL, and improve the production of IFN-gamma and TNF-alpha, thus enhance the anti-leukemia activity of CTL. Nevertheless, there is no effect of IL-21 on the function of mature DCs. These data indicate that IL-21 has a potential clinical value in the enhancement of anti-leukemia immunotherapy.

Analyses of surgical treatment and prognosis in gastric stump cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of surgical treatment and the prognosis in gastric stump cancer patients.</p><p><b>METHODS</b>Between June 1994 and March 2004, 692 patients underwent radical operation for gastric cancer in our department. Among them, 22 cases were gastric stump cancer. Their surgical treatments, lymph node metastasis and survival were analyzed retrospectively.</p><p><b>RESULTS</b>Gastric stump cancer accounted for 3.2 % of all the gastric cancer cases in the same period. There were 4 cases of stage I, 2 cases of stage II, 6 cases of stage III and 10 cases of stage IIII respectively. Radical excision was 77.3% and combined evisceration was 50.0%. Total gastrectomy were performed in 21 cases and exploratory laparotomy in 1 case. Digestive tract was reconstructed with Roux-en-Y pattern after total gastrectomy. Radical gastric stump cancer excisions were finished with abdominal incision in 18 cases and with thoraco-abdominal incision in 4 cases. Lymph node metastasis rate was 63.6 %, including pN(0) 8 cases, pN(1) 6 cases, pN(2) 7 cases and pN(3) 1 case respectively. Average survival time was(80.2+/-17.2) months in stage I( and II( gastric remnant cancer; average survival time was(31.2+/-9.2) months in stage III( gastric remnant cancer, average survival time was (23.6+/-6.1) months in stage IIII( gastric remnant cancer, which were significantly different(all P<0.05). Between palliative operation group and standard radical excision, extended radical excision groups, well-moderate differentiated and poor differentiated adenocarcinoma groups, lymph node metastasis positive and negative groups, the differences were all significant.</p><p><b>CONCLUSIONS</b>Total gastrectomy and D(2) lymph node dissection are imperative for radical excision of gastric remnant cancer. On this base, extended lymphectomy and combined evisceration should be performed appropriately. Tumor stage, procedure pattern, lymph node metastasis and tumor differentiation affect the prognosis of patients with gastric stump cancer.</p>

Extracellular matrix accumulation and expression of gelatinases and their tissue inhibitors in a mechanically unloaded heart model / 中国应用生理学杂志

<p><b>AIM</b>To investigate the relationship between the expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 and ECM accumulation in rat left ventricle in a mechanical unloaded heart model.</p><p><b>METHODS</b>12-week-old male Lewis rats were subjected to abdominal heterotopic heart transplantation to achieve pressure and volume unloading(mechanical unloading). Age and sex matched in situ heart of Lewis rats were used as control. Collagen volume fraction(CVF) was analyzed by picrosiris-red staining plus polarized microscopy. MMP-2 and -9 gelatinolytic activity were measured by gelatin-zymography. mRNA level of MMP-2, MMP-9, TIMP-1 and TIMP-2 were measured by real-time quantitative PCR. TIMP-1 and TIMP-2 protein level were measured by immunoblotting.</p><p><b>RESULTS</b>Myocardial cross-sectional area of transplanted heart was significantly reduced, and accompanied by excessive ECM deposition (CVF 5.22% +/- 1.6% vs. 2.21% +/- 0.9%, P < 0.05) compared to in situ heart. MMP-2 and MMP-9 activity were significantly increased, as well as mRNA level of MMP-2, MMP-9, TIMP-1 and TIMP-2 compared to in situ heart. TIMP-1 and TIMP-2 protein level in mechanically unloaded heart were significantly upregulated compared to in situ heart, especially for TIMP-1.</p><p><b>CONCLUSION</b>Mechanical unloading of left ventricle may lead to excessive ECM deposition, accompanied by imbalance between MMPs and TIMPs system, especially the upregulation of TIMPs.</p>