Mechanism of cinnamic aldehyde-inducing apoptosis of chronic myeloid leukemic cells in vitro / 中国实验血液学杂志

The aim of this study was to investigate the apoptosis-inducing effect of cinnamic aldehyde (CA) on chronic myeloid leukemic (CML) cells and its mechanism. K562 cells and primary bone marrow mononuclear cells (MNC) from patients with CML were treated by various concentrations of CA. Flow cytometry was employed to measure the apoptosis of K562 cells and primary CML bone marrow MNC. Western blot was used to determine the expression of C-MYC and the phosphorylation of CrkL in K562 cells, and real-time polymerase chain reaction (real-time PCR) was used to quantify the expression of BCR-ABL mRNA in K562 cells. The results indicated that CA induced the apoptosis of K562 cells in a time- and dose-dependent manner. CA induced apoptosis of CML MNC dose-dependently. CA inhibited the expression of BCR-ABL mRNA and C-MYC, reduced CrkL phosphorylation levels in K562 cells. It is concluded that CA induces apoptosis of CML cells in vitro. Down-regulation of the expression and function of BCR-ABL may be one of its most important anti-leukemia mechanisms.

Proliferative response of CD62L(-)/CD62L(+) T cells in mice to antigen stimulation / 中国实验血液学杂志

The aim of this study was to investigate the ability of CD62L(-)/CD62L(+) T cells responding to antigens so as to explore the role of CD62L in GVHD. The T cells were stimulated by DCs-presenting specific antigen hCD4; the expression of CD62L on surface of T cells was detected by flow cytometry, the CD62(-) T/CD62L(+) T cells were gained by sorting selection method, then were stimulated with a new antigen (freeze-thaw antigen of EL4 cells), and the proliferation activity of CD62L(-) and CD62L(+) T cells was detected by flow cytometry after stimulating with different antigens at twice. The results showed that after stimulating with antigen, the expression level of CD62L(+) on surface of T cells was down-regulated from 60.34% to 36.75%, and the expression of CD62L(-) on surface of T cells increased from 30.04% to 63.15%. Compared with cells counts in area with high CFSE (M1), the average percentages of CD62L(+) T cells under stimulating with specific antigen (hCD4) on days 1 and 6 were (87.88 +/- 1.08)% and (86.88 +/- 1.46)% respectively (p > 0.05), and the average percentages of CD62L(+) T cells after stimulating with specific antigen on days 1 and 6 were (84.52 +/- 2.73)% and (84.21 +/- 2.33)% respectively (p > 0.05), there was no significant difference between two T cell groups on days 1 and 6. After stimulating with non-specific antigen, the average percentages of CD62L(+) T cells in MI area on days 1 and 6 were (76.49 +/- 2.41)% and (22.25 +/- 3.29)% respectively (p < 0.001), the average percentages of CD62l(-) T cells under conditions mentioned above were (77.35 +/- 3.82)% and (74.76 +/- 3.90)% respectively (p < 0.05). The results indicated that the proliferation activity of CD62L(+) T cells appeared, and the proliferation of CD62L(+) T cells did not occur. It is concluded that the CD62L(-) T/CD62l(+) T cells are gained in vitro, no proliferation reaction occurred in these two kinds of cells under stimulation of specific antigen, but the mitotic proliferation occur in CD62L(+) T cells under stimulation of nonspecific antigen, while no this reaction exists in CD62L(-) T cells under this condition. These results provide a new thinking to obtain the CD62L(-) T cells in vitro, to selectively eliminate the CD62L(+) T cells from graft and to decrease occurrence of GVHD in clinical practice.

Subjective and objective clinical outcome assessment on chronic rhinosinusitis following endoscopic sinus surgery / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To assess the subjective and objective outcomes of chronic rhinosinusitis (CRS) following endoscopic sinus surgery (ESS) and establish an assessment system of outcome with ease of application clinically.</p><p><b>METHODS</b>A prospective cohort study was conducted to survey and assess the outcomes of 120 consecutive CRS patients undergoing endoscopic sinus surgery at 12 months after operation. The subjective and objective measures comprised symptom by visual analog scale (VAS), health-related quality of life by medical outcome study short-form 36-items (SF-36) and sino-nasal outcome test-20 (SNOT-20) scales, endoscopic appearance, mucociliary function, and histological findings. The differences of subjective and objective assessments before and after operation were compared by t-test and Chi-Square test and the correlations between the parameters above were analyzed by Spearman correlation analysis.</p><p><b>RESULTS</b>At 12 months after operation, the patients' total scores by VAS, SF-36 and SNOT-20 scales improved significantly beyond the preoperative survey (P < 0.01); there were 85.96%, 77.19% and 83.33% patients with the scores respectively superior to that of preoperation, of which 72.28% subjects benefited simultaneously from these parameters; and a significant correlation was observed among them before and after operation (P < 0.01) where SNOT-20 showed a more compatibility than the other two. At 12 months after operation, the patients' total scores of endoscopic appearance, mucociliary function, and histological findings significantly improved beyond the preoperative evaluation (P < 0.05); there were 86.84% , 86.81% and 75.57% patients with the scores respectively superior to that of preoperation, of which 71.85% subjects benefited simultaneously from these parameters; and a significant correlation was observed among them before and after operation (P < 0.05) where endoscopic appearance showed a more compatibility than the other two. At 12 months after operation, 74.56% patients showed an accordant improving or worsening outcome evaluated by SNOT-20 and endoscopic appearance, while 25.44% ones represented inverse endings, of which patients with comorbidity of nasal polyps more easily demonstrated this tendency significantly (P < 0.05). No significant correlation existed between the scores of SNOT-20 and endoscopic appearances both in preoperation and in postoperation (P > 0.05), but the total scores of the anterior 10-item, excluding the posterior 10-item, of SNOT-20 inventory was found significantly correlated with the quantitative appearances on nasal endoscopy throughout (0.18 < or = 0.42, P < 0.05).</p><p><b>CONCLUSIONS</b>Administration of ESS can effectively improve the outcomes of CRS patients including symptom, health-related quality of life, endoscopic appearance, mucociliary function, and histological findings. A subjectively and objectively measured assessment system with tenseness, trustiness, reasonableness, and effectiveness and with ease of application clinically is established on the basis of SNOT-20 and endoscopic appearance evaluation for outcome research.</p>

Quality of life outcome of patients with chronic rhinosinusitis and nasal polyposis after endoscopic sinus surgery and its influencing factors / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the quality of life (QOL) outcome of patients with chronic rhinosinusitis (CRS) after endoscopic sinus surgery (ESS) and its influencing factors.</p><p><b>METHODS</b>prospective trial was conducted to survey the QOL status of 120 CRS patients undergoing ESS, in contrast that of 200 healthy individuals passing health examination, at the baseline and at 12-months after operation by Medical Outcomes Study Short Form-36 (SF-36) and Sino-Nasal Outcome Test-20 (SNOT-20). QOL changes and its influencing factors were analyzed statistically.</p><p><b>RESULTS</b>(1) By the assessment of SF-36, the scores of 6 domains were less than that of healthy individuals preoperatively (P < 0.01). After 6 months, the scores of these domains resumed normal level and the proportion of scores also restored normally (P > 0.05). (2) By the assessment of SNOT-20, the total scores of 20 items and 5 most important items of patients were more than that of healthy objects (P < 0.01). After 9 and 12 months, the former and latter returned to normal, respectively (P > 0.05). In 12 months setting, the proportion of scores also restored normally (P > 0.05). (3) According to the survey of SNOT-20, we concluded the following equation: convalescent time (months) = 39--(normal scores/preoperative scores) x 50, by which the time of coming back to normal QOL status can be computed. (4) By analysis of Logistic Regression, residence in city or country, course of disease, extension of diseased sinus, and coexistence of nasal polyposis or not were correlated to the preoperative QOL scores; working environments, surgical extension, and preoperative scores of QOL were correlated to the score difference between pre and post operation.</p><p><b>CONCLUSIONS</b>CRS patients undergoing ESS could obtain entirely normal QOL status at 12 months postoperatively, so we suggest that the essential follow-up period should last at least one year. The risk factors influencing patients QOL status preoperatively includes residence in country, longer course of disease, more extension of diseased sinus, and coexistence of nasal polyposis. The risk factors hindering the improvement of QOL status postoperatively includes exposure to indoor working environments, insufficient surgical extension, and lower preoperative QOL scores.</p>

Effect of WISp39 on proliferation, cell cycle and apoptosis of U937 cells / 中国实验血液学杂志

To investigate the effect of a novel p21-modulating protein WISp39 on proliferation, apoptosis and cell cycle of leukemia cells, the plasmid pLenti6/V5-WISp39 was constructed and transfected into the human myelocytic leukemia cell line-U937 cells. The expression of WISp39 was detected by real-time PCR at 48 hours after transfection, proliferation of U937 cells assayed by CCK-8, apoptosis and cell cycle were determined by flow cytometry. The results showed that plasmid pLenti6/V5-WISp39 could readily enhance the expression of WISp39 in U937 cells. A significant growth inhibition (37.6%) was observed in cells tranfected with pLenti6/V5-WISp39, while the control plasmid pLenti6/V5-lacZ showed little effect on U937 growth. Further analysis revealed that pLenti6/V5-WISp39 did not show obvious apoptosis induction effect, but it could really regulate U937 proliferation via cell cycle modulation. Compared with pLenti6/V5-lacZ, pLenti6/V5-WISp39 resulted in increase of cells in G(0)/G(1) phase by 10% at 48 hours after transfection. It is concluded that the WISp39 gene has no significant apoptosis induction effect on leukemic cells, but it can increase cells at G(0)/G(1) phase via effect on cell cycle, thus inhibiting the U937 proliferation. This result means WISp39 gene can act as a negative modulator on tumour cells.

Expression and role of toll-like receptors in U937 cells / 中国实验血液学杂志

The aim of study was to explore the potential application of targeting at Toll-like receptors (TLRs) in the immunotherapy of acute myelocytic leukemia, and to investigate the expression of TLR and the effects of TLR 8 agonist ssRNA40/LyoVec on proliferation, apoptosis and cell cycle of U937 cells. The expression of TLR 1 - 9 in U937 cells was detected by using reverse transcription polymerase chain reaction (RT-PCR) and the expression of TLR 8 was assayed by flow cytometry (FCM). The effect of TLR 8 agonist, ssRNA40/LyoVec, at different concentrations on U937 cells proliferation was evaluated by CCK-8, apoptosis and cell cycle were detected by FCM. The results showed that U937 cells expressed TLR 1 - 9. TLR 8 agonist ssRNA40/LyoVec could inhibit the growth of U937 cells both in time-and dose-dependent manner and the inhibitory rate could reach 70%. It also increased the percentage of cells in G(0)/G(1) phase. There was no significant difference in percentage of apoptotic cells between control and treated groups. It is concluded that TLRs including TLR 1 - 9 express on U937 cells and TLR 8 agonist ssRNA40/LyoVec may be able to inhibit the growth of U937 cells, arrest the cells in G(0)/G(1) phase, but have no effect of promoting apoptosis.

Effect of CCL23/myeloid progenitor inhibitory factor 1 (MPIF-1) on the proliferation, apoptosis and differentiation of U937 cells / 中国实验血液学杂志

CCL23 is a human CC chemokine with potential suppression effects on both human and murine myeloid progenitor cells both in vitro and in vivo, and only expressed and released by dendritic cells differentiated from monocytes in blood cells. However, recent study has shown that CCL23 was over-expressed in bone marrow and peripheral blood cells from pediatric patients with acute myeloid leukemia (AML). In order to investigate the effects of CCL23 on the development, therapy and prognosis of leukemia, the U937 cells, a leukemic cell strain, were adopted and cultured with rhCCL23 for 72 hours. The cell proliferation and apoptosis rate were detected by Cell Counting Kit-8 and FITC-AnnexinV/PI respectively; the morphologic changes and the expression of CCR1 (the only receptor of CCL23 known by now) were observed during the differentiation process. The results showed that no obvious effect on the proliferation, apoptosis and differentiation of U937 was found by using CCL23 alone (P > 0.05), but cultured in combination with CCL23 and PMA, the differentiation of U937 cells were promoted remarkably, during which the CCR1 expression increased (P < 0.05). It is concluded that CCL23 alone did not inhibit the proliferation and differentiation of U937, while its use in combination with PMA may possess synergistic effect on inducting differentiation of U937 through the increase of receptor CCR1 expression.

Endoscopic evaluation of mucous membrane inflammation in chronic rhinosinusitis and analysis of correlated factors / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>The purpose of this article is to establish an endoscopic score system for quantitative evaluation of the inflammation of mucous membrane in patients with chronic rhinosinusitis (CRS) , and to investigate the correlation of this system with a variety of clinical factors.</p><p><b>METHODS</b>A set of score system was constructed based on anatomic configuration, status of mucous membrane and nasal secretion to evaluate quantificationally the severity of inflammation of CRS. The clinical correlation of this system was studied prospectively in 60 CRS patients, with a variety of clinical factors which included age, duration of disease, previous recurrence and the years from recent recurrence, atopy, serum total immunoglobulin E (TIgE), serum eosinophil cationic protein (ECP), the count of blood eosinophil, the count of tissue inflammatory cell, the extension of CRS indicated by CT, smoking, concomitant chronic inflammation in lower respiratory tract. All above factors were analyzed statistically with the endoscopic score by Pearson correlation and multi-factor linear regression analysis.</p><p><b>RESULTS</b>In pearson analysis, the correlative factors with the evaluated score included age (x1, r = - 0.310, P = 0.016), the extension of disease (x2, r = 0. 810, P < 0.0005), recurrence (x3, r = 0.408, P = 0.001), eosinophil of nasal tissue (x4, r = 0.279, P = 0. 031), duration of disease (x5, r = 0.536, P < 0.0005), concurrent nasal polyps (r = 0.549, P < 0.0005), plasm cell (r = 0. 317, P = 0.014) and years from the recent recurrence (r = 0.385, P = 0.002). In multi-factor linear regression, the five independent predictive factors were recurrence, age, extension of disease, tissue eosinophils, years of disease. The regressive equation is y = 10.148 - 0.152 (x1) + 2.250 (x2) + 3.348 (x3) + 1.233 (x4) + 0.270 (x5).</p><p><b>CONCLUSIONS</b>Appropriate score system by nasal endoscopy is feasible to evaluate quantificationally the degree of inflammation of CRS; being appropriately modified, it is even able to reveal the underlying histological behavior finely.</p>