Application of next-generation sequencing in research and personalized treatment of glioma / 第二军医大学学报

Glioma is a highly lethal malignancy and multiple challenges remain for it in the genomic study and clinical treatment. Next-generation sequencing (NGS) holds many advantages, and NGS can comprehensively analyze the whole genome, exome, transcriptome and epigenome to deepen the understanding of tumor genomics; and now, it has been successfully applied in the clinical study of some tumors, especially in personalized treatment for these tumors. At present, NGS has also been a hot in glioma research to clarify the pathogenesis of glioma, so as to draw up a personalized treatment method to benefit the patients.

Inhibitory effects of parthenolide on the activity of NF-κB in multiple myeloma via targeting TRAF6 / 华中科技大学学报（医学）（英德文版）

This study examined the mechanism of the inhibitory effect of parthenolide (PTL) on the activity of NF-κB in multiple myeloma (MM). Human multiple myeloma cell line RPMI 8226 cells were treated with or without different concentrations of PTL for various time periods, and then MTT assay was used to detect cell proliferation. Cell cycle and apoptosis were flow cytometrically detected. The level of protein ubiquitination was determined by using immunoprecipitation. Western blotting was employed to measure the level of total protein ubiquitination, the expression of IκB-α in cell plasma and the content of p65 in nucleus. The content of p65 in nucleus before and after PTL treatment was also examined with immunofluorescence. Exposure of RPMI 8226 cells to PTL attenuated the level of ubiquitinated Nemo, increased the expression of IκB-α and reduced the level of p65 in nucleus, finally leading to the decrease of the activity of NF-κB. PTL inhibited cell proliferation, induced apoptosis and blocked cell cycle. Furthermore, the levels of ubiquitinated tumor necrosis factor receptor-associated factor 6 (TRAF6) and total proteins were decreased after PTL treatment. By using Autodock software package, we predicted that PTL could bind to TRAF6 directly and tightly. Taken together, our findings suggest that PTL inhibits the activation of NF-κB signaling pathway via directly binding with TRAF6, thereby suppressing MM cell proliferation and inducing apoptosis.

Inhibitory effects of parthenolide on the activity of NF-κB in multiple myeloma via targeting TRAF6 / 华中科技大学学报（医学）（英德文版）

This study examined the mechanism of the inhibitory effect of parthenolide (PTL) on the activity of NF-κB in multiple myeloma (MM). Human multiple myeloma cell line RPMI 8226 cells were treated with or without different concentrations of PTL for various time periods, and then MTT assay was used to detect cell proliferation. Cell cycle and apoptosis were flow cytometrically detected. The level of protein ubiquitination was determined by using immunoprecipitation. Western blotting was employed to measure the level of total protein ubiquitination, the expression of IκB-α in cell plasma and the content of p65 in nucleus. The content of p65 in nucleus before and after PTL treatment was also examined with immunofluorescence. Exposure of RPMI 8226 cells to PTL attenuated the level of ubiquitinated Nemo, increased the expression of IκB-α and reduced the level of p65 in nucleus, finally leading to the decrease of the activity of NF-κB. PTL inhibited cell proliferation, induced apoptosis and blocked cell cycle. Furthermore, the levels of ubiquitinated tumor necrosis factor receptor-associated factor 6 (TRAF6) and total proteins were decreased after PTL treatment. By using Autodock software package, we predicted that PTL could bind to TRAF6 directly and tightly. Taken together, our findings suggest that PTL inhibits the activation of NF-κB signaling pathway via directly binding with TRAF6, thereby suppressing MM cell proliferation and inducing apoptosis.

Amplification of functional myeloid-derived suppressor cells during stem cell mobilization induced by granulocyte colony-stimulation-factor / 华中科技大学学报（医学）（英德文版）

The effects of granulocyte colony-stimulation-factor (G-CSF) on stem cell mobilization and its impact on the amplification of myeloid-derived suppressor cells (MDSCs) of donor mice were examined. A mouse model of stem cell mobilization was established by consecutive subcutaneous injection of 100 μg/kg G-CSF for 5 days. The blood from the donor mice was routinely examined during mobilization. Stem cells and MDSCs were analyzed by flow cytometry. The immunosuppressive molecules derived from MDSCs in serum and spleen, including hydrogen dioxide (H2O2) and nitric oxide (NO), and the activity of nitric oxide synthase (NOS) were determined during the mobilization. Apoptosis of T lymphocytes was assessed by using Annexin-V/PI. During stem cell mobilization, the number of lymphocytes and white blood cells in the peripheral blood was increased, and peaked on the 4th day. The number of stem cells in G-CSF-treated mice was significantly greater than that in controls (P<0.01). The expansions of MSDCs were also observed after G-CSF mobilization, with a more notable rate of growth in the peripheral blood than in the spleen. The activity of NOS and the production of NO were increased in the donor mice, and the serum H2O2 levels were approximately 4-fold greater than the controls. Consequently, apoptosis of T lymphocytes was increased and showed a positive correlation with the elevated percentage of MDSCs. It was concluded that G-CSF could provide sufficient peripheral blood stem cells for transplantation. Exogenous administration of G-CSF caused the accumulation of MDSCs in the peripheral blood and the spleen, which could lead to apoptosis of T lymphocytes and may offer a new strategy for the prevention and treatment of graft versus host disease.

Amplification of functional myeloid-derived suppressor cells during stem cell mobilization induced by granulocyte colony-stimulation-factor / 华中科技大学学报（医学）（英德文版）

The effects of granulocyte colony-stimulation-factor (G-CSF) on stem cell mobilization and its impact on the amplification of myeloid-derived suppressor cells (MDSCs) of donor mice were examined. A mouse model of stem cell mobilization was established by consecutive subcutaneous injection of 100 μg/kg G-CSF for 5 days. The blood from the donor mice was routinely examined during mobilization. Stem cells and MDSCs were analyzed by flow cytometry. The immunosuppressive molecules derived from MDSCs in serum and spleen, including hydrogen dioxide (H2O2) and nitric oxide (NO), and the activity of nitric oxide synthase (NOS) were determined during the mobilization. Apoptosis of T lymphocytes was assessed by using Annexin-V/PI. During stem cell mobilization, the number of lymphocytes and white blood cells in the peripheral blood was increased, and peaked on the 4th day. The number of stem cells in G-CSF-treated mice was significantly greater than that in controls (P<0.01). The expansions of MSDCs were also observed after G-CSF mobilization, with a more notable rate of growth in the peripheral blood than in the spleen. The activity of NOS and the production of NO were increased in the donor mice, and the serum H2O2 levels were approximately 4-fold greater than the controls. Consequently, apoptosis of T lymphocytes was increased and showed a positive correlation with the elevated percentage of MDSCs. It was concluded that G-CSF could provide sufficient peripheral blood stem cells for transplantation. Exogenous administration of G-CSF caused the accumulation of MDSCs in the peripheral blood and the spleen, which could lead to apoptosis of T lymphocytes and may offer a new strategy for the prevention and treatment of graft versus host disease.

Treatment of varicocele infertility men patients of different Chinese medical syndrome types by integrative medicine treatment selection: a primary research / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the optimal treatment selection for treating varicocele (VC) male infertility patients accompanied with oligozoospermia or azoospermia of different Chinese medical syndrome types by comparing the efficacies of integrative medicine.</p><p><b>METHODS</b>One hundred and twenty male infertility patients with VC accompanied with oligozoospermia or azoospermia were assigned to Chinese medical treatment group (A) and the surgical group (B), each consisting of three Chinese medical syndrome types, i.e., damp-heat stagnation syndrome (DHSS), Shen-deficiency blood stasis syndrome (SDBSS), and blood stasis stagnation syndrome (BSSS), 20 in each group. Corresponding Chinese medical treatment was administered to those in Group A, C, and E, while microscopic ligation of internal vena spermatic was administered to those in Group B, D, and F. The routine analysis of semen, biochemical analysis of seminal plasma, and serum sex hormones (prolactin, testosterone, follicle stimulating hormone, luteinizing hormone, and estradiol) were performed before treatment and by the end of the 24th week after treatment.</p><p><b>RESULTS</b>Totally 18 patients' spouses were pregnant. Of them, 1 in Group A of DHSS (abbreviated as Group A), 3 in Group B of DHSS (abbreviated as Group B), 4 in Group A of SDB-SS (abbreviated as Group C), 5 in Group A of SDBSS (abbreviated as Group D), 1 in Group A of BSSS (abbreviated as Group E), and 4 in Group B of BSSS (abbreviated as Group F). After 24-week treatment, the sperm concentration, class a sperm percentage, class a + b sperm percentage, the motility rate, the seminal plasma of fructose density, and the seminal plasma neutral alpha-glucosidase were more significantly improved in Group B, C, D and F, when compared with the same group before treatment (P <0. 01, P <0. 05). There was no statistical difference in the aforesaid indices between before and after treatment in Group A and Group E (P >0.05). As for the improvement percentage of seminal routine indices, the difference of the seminal plasma of fructose density, and the difference of seminal plasma neutral alpha-glucosidase between before and after treatment in the same Chinese medical syndrome types, better effects were obtained in Group B than in Group A (P <0.01), and better effects were obtained in Group F than in Group E (P <0.01). There was no statistical difference between Group C and D (P >0.05). There was no statistical difference in the 5 items of sex hormones in each group between before and after treatment (P >0.05).</p><p><b>CONCLUSIONS</b>Surgical treatment could effectively improve the semen quality for male infertility VC patients accompanied with oligozoospermia or azoospermia. Of them, Chinese medical treatment could be recommended to those of SDBSS who would not receive surgical treatment.</p>

Sexual function inspection room for special penile insertion disorder / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine the role of the sexual function inspection room in the treatment of patients with special penile insertion disorder.</p><p><b>METHODS</b>This study included 6 couples received at the Andrology Clinic of Shanghai Renji Hospital from September, 2010 to February, 2011, who complained of being unable to accomplish sexual intercourse, all due to the husbands' abnormal penile erection. The couples had tried to make love 3 to 6 times after getting married, but never succeeded. Consequently the wives were planning to divorce, unwilling to attempt sexual activity again. We performed examinations and sexual education for the patients in the sexual function inspection room of Shanghai Institute of Andrology. The inspection room consisted of an inner and an outer section separated by a one-way transparent glass, through which what happened in the outer section could be observed from the inner section. The husband was given 20 mg of tadalafil to induce penile erection, followed by Doppler ultrasonographic determination of penile hemodynamics. Meanwhile, the wife was allowed to observe her husband's penile erection through the glass to get sensory stimuli and relieve the mental burden. In the end, the doctor advised the couple to try sexual intercourse.</p><p><b>RESULTS</b>In nature, the 6 wives were all impatient, while their husbands were all introverted. Doppler ultrasonography displayed normal penile hemodynamics. After sensory stimulation, the wives gained confidence in their husbands' erectile function. A one month follow-up visit showed that all the 6 couples could perform sexual intercourse normally.</p><p><b>CONCLUSION</b>This sexual function inspection room can help doctors and wives to observe the status of the patient's penile erection, and meanwhile disburden the mind of the wives by giving them sensory stimuli, which contributes much to successful attempt at sexual intercourse.</p>

Leptin level in azoospermic patients and its clinical value / 中华男科学杂志

<p><b>OBJECTIVE</b>To detect the levels of seminal plasma leptin (SPL) and serum leptin (SL) in patients with azoospermia, and to explore the methods of using SPL and SL alone or the combination of SPL, SL and follicle stimulating hormone (FSH) for the differential diagnosis of obstructive azoospermia (OA) and non-obstructive azoospermia (NOA).</p><p><b>METHODS</b>We enrolled in this study 45 patients with diagnosed OA, 41 with unexplained NOA and 30 men with normal semen parameters as controls. The azoospermia patients underwent percutaneous aspiration from the epididymis (PESA) or aspiration/extraction from the testis (TESA/TESE), and all the subjects were detected for the levels of serum FSH, SPL and SL. Individual and multiple indexes were evaluated by Fisher's discriminant analysis combined with ROC curve analysis.</p><p><b>RESULTS</b>There were no significant differences in the body mass index (BMI) among the three groups. Compared with the normal control, the OA patients showed an obviously elevated level of SPL (P = 0.048), and the NOA patients remarkably increased levels of FSH (P = 0.000), SL (P = 0.000) and SPL (P = 0.000). In comparison with the OA group, the levels of FSH (P = 0.000), SL (P = 0.006) and SPL (P = 0.033) were significantly increased in the NOA group. For the differential diagnosis of OA and NOA, the areas under the ROC curve of SPL and SL were 0.658 (P = 0.014) and 0.702 (P = 0.002) , respectively, both significantly greater than 0.5, while that of the combination of SPL, SL and FSH was the greatest (0.953). In addition, with 0.026 x SPL +0.05 x SL +0.106 x FSH -2.197 as the combined indicator value and -0.289 as the cut-off value (> or = cut-off value for NOA), the sensitivity and specificity of the combination were 0.878 and 0.902, respectively, both reached the maximum.</p><p><b>CONCLUSION</b>Both the levels of SPL and SL are valuable for the differential diagnosis of OA and NOA, but the joint consideration of SPL, SL and FSH may provide better indicators.</p>

Prevalence and related risk factors of peripheral arterial disease in elderly patients with type 2 diabetes in Wuhan, Central China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The investigations of prevalence and risk factors of peripheral arterial disease (PAD) in type 2 diabetic patients have been carried out in many countries and regions, except for Central China. In this study, we determined the prevalence of PAD in type 2 diabetic patients and the related factors that gave rise to increasing of the risk of PAD development in Wuhan, China.</p><p><b>METHODS</b>The study enrolled 2010 patients aged 60 years and older who were regularly visiting the Central Hospital of Wuhan that is a public hospital from 2005 to 2010, where all residents of the city were offered the medical services. PAD was defined as an ankle-brachial index < 0.90 in either leg. To evaluate the role of various risk factors in PAD development, uniformed interviews, clinical examinations and laboratory investigation of all of participants were performed in this study. The correlation between potential risk factors and PAD was analyzed.</p><p><b>RESULTS</b>In Wuhan, the prevalence rate of PAD was 24.1% in elderly diabetic patients. Totally, 291 patients with PAD had at least one weak but not absent dorsalis pedis pulse in both feet, while 541 patients without PAD showed this way. At least one absent dorsalis pedis pulse was found in 192 patients with PAD as well as 10 patients without PAD. The results of multivariate regression analysis suggested that the age, smoking history, hypertension, diabetic neuropathy and macroangiopathy gave rise to significant increase of PAD development in type 2 diabetic diseases.</p><p><b>CONCLUSIONS</b>The prevalence of PAD in elderly patients with type 2 diabetes in Wuhan was close to the prevalence that was reported in other regions of China and other Asian countries. Control of the related risk factors and early diagnosis of PAD may play a role in PAD prevention and improving prognosis.</p>

Autologous bone marrow stem cell transplantation for the treatment of type 2 diabetes mellitus / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Autologous peripheral stem cell transplantation was first reported in 2007 to treat type 1 diabetes mellitus (DM) and achieved encouraging effect, but whether similar outcome can be achieved in type 2 DM is not well demonstrated. The objective of this study was to determine the effect of combination of autologous bone marrow stem cell transplantation (BMT) and hyperbaric oxygen treatment on type 2 DM.</p><p><b>METHODS</b>The study involved 31 patients with type 2 DM (aged 33 to 62 years) from January 2009 to January 2011 in the Central Hospital of Wuhan, China. Clinical variables (body mass index, duration of DM, insulin requirement, oral hypoglycemic drugs, time free from insulin, time free from oral drugs) and laboratory variables (hemoglobin A1c (HbA1c)), mononuclear cells infused, and C-peptide in four time points) were assessed. Purified bone marrow stem cells were infused into major pancreatic arteries. Follow-up was performed at the 30, 90, 180, 360, 540 and 720 days (mean 321 days) after BMT.</p><p><b>RESULTS</b>Mean HbA1c values showed a significant reduction during follow-up in all patients after BMT. It decreased by more than 1.5% (from 8.7% to 7.1%) as quickly as at 30 days after BMT. Afterwards mean HbA1c fluctuated between plus or minus 0.5% until 24 months rather than declined continuously. At 90 days after the combined therapy C-peptide increased significantly compared with baseline (P < 0.0001). But in other time points C-peptide was similar with baseline data (P > 0.3). All patients had insulin and/or oral hypoglycemic drugs reduced to different levels. The dose of insulin of 7 patients (7/26, 27%) reduced for a period of time after BMT.</p><p><b>CONCLUSIONS</b>Combined therapy of intrapancreatic BMT and hyperbaric oxygen treatment can improve glucose control and reduce the dose of insulin and/or oral hypoglycemic drugs in type 2 DM patients, but it only improve pancreatic β-cell function transiently. Further randomized controlled clinical trials involved more patients will be required to confirm these findings and the mechanism needs to be illustrated deeply.</p>

Sexual and reproductive health service needs of university/college students: updates from a survey in Shanghai, China / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To promote the provision of reproductive health services to young people by exploring the attitudes and perceptions of university students in Shanghai, China, toward reproductive health.</p><p><b>METHODS</b>From July 2004 to May 2006, 5 243 students from 14 universities in Shanghai took part in our survey. Topics covered the demands of reproductive health-care services, attitudes towards and experience with sex, exposure to pornographic material, and knowledge on sexual health and sexually transmitted infections (STIs)/AIDS.</p><p><b>RESULTS</b>Of the 5 067 students who provided valid answer sheets, 50.05% were female and 49.95% were male, 14.86% were medical students, and 85.14% had non-medical backgrounds. A total of 38.4% of respondents had received reproductive health education previously. The majority of students supported school-based reproductive health education, and also acquired information about sex predominantly from books, schoolmates, and the Internet. Premarital sexual behavior was opposed by 17.7% of survey participants, and 37.5% could identify all the three types of STIs listed in the questionnaire. Although 83.7% knew how HIV is transmitted, only 55.7% knew when to use a condom and 57.8% knew that the use of condoms could reduce the risk of HIV infection.</p><p><b>CONCLUSION</b>The reproductive health service is lagging behind current attitudes and demands of university students. Although students' attitudes towards sexual matters are liberal, their knowledge about reproductive health and STIs/AIDS is still limited. It is therefore necessary to provide effective and confidential reproductive health services to young people.</p>

The feeder layer of human embryonic fibroblasts supports the growth of human spermatogonial stem cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the methods and conditions for the isolation, purification and culture of human spermatogonial stem cells (SSCs) on the feeder layer cells of human embryonic fibroblasts (hEFs).</p><p><b>METHODS</b>SSCs isolated and purified from normal human fetal testicular tissues by sequential two-step enzyme digestion and Percoll uncontinuous density gradient centrifugation were cultured on the feeder layer cells of hEFs isolated from 5-9 weeks old human embryos. The surface markers SSEA-1 and OCT4 of the SSCs were detected by immunohistochemistry; the alkaline phosphatase (AKP) activity of the SSC clones measured; and the expressions of the SSC-related genes determined by RT-PCR.</p><p><b>RESULTS</b>SSCs survived, proliferated and formed colonies on the feeder layers, and the colonies were highly positive for SSEA-1 and OCT4, with strong AKP activity and high expressions of the SSC-related genes.</p><p><b>CONCLUSION</b>The feeder layer of hEFs supports the growth of human spermatogonial stem cells.</p>

Indoleamine 2, 3-dioxygenase activity in acute myeloid leukemia cells contributing to tumor immune escape / 中国实验血液学杂志

This study was aimed to investigate the mechanism of indoleamine 2, 3-dioxygenase (IDO) activity in acute myeloid leukemia cells contributing to tumor immune escape. Myeloid leukemia cells were isolated from bone marrow of 23 patients with acute myeloid leukemia (AML) and IDO expression was detected by immunochemistry and RT-PCR methods. Then mixed lymphocyte reaction (MLR) of one way was carried out, leukemia cells were used as stimulating cells and T-lymphocytes were used as reactive cells in culture with or without 1-MT. T-lymphocyte proliferation rate was determined by MTT assay and IDO activity in supernatant of MLR was detected by high-performance liquid chromatography (HPLC). The results showed that IDO expression was found in 17 out of 23 cases of acute myeloid leukemia cells; IDO enzyme activity in leukemia cells inhibited T-lymphocyte proliferation in MLR cultures. It is concluded that IDO activity expressing in leukemia cells can suppress T-lymphocyte proliferation responses, which may be contributing to tumor immune escape.

Comparative Study of Mesenchymal Stem Cells Derived From Human Umbilical Cord Vein and Adult Bone Marrow / 中国生物工程杂志

Mesenchymal stem cells (MSCs) are promising candidates to develop new cell-based therapeutic strategies and bone marrow represents the main source of MSCs for experimental and clinical application currently. How to obtain enough MSCs is the biggest challenge faced by the experimental and clinical application. The difference of morphology, growth pattern, immunophenotype and multilineage differentiation of MSCs derived from human umbilical cord vein and their multiple differentiation capacity were investigated. The results showed that MSCs derived from human umbilical cord vein and normal adult bone marrow were similar in biological characteristics. They not only grew in fibroblast-like cells mode but had great expansion potency and multilineage differentiation ability in vitro. The MSCs derived from human umbilical cord vein may be an excellent alternative source for experimental and clinical application.

Survey on reproductive health status of Shanghai college students / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore reproductive health condition of college students in Shanghai and provide scientific basis for reproductive health education.</p><p><b>METHODS</b>From July 2004 to March 2005, 1192 college students (majors including medicine, science and engineering, arts and culture; 618 medical students and 549 non-medical; 555 male and 612 female) from seven time-day colleges in Shanghai were investigated with a reproductive health questionnaire involving sex. The questionnaire was constituted by 72 topics, involving the need of education, sex attitude, sex videos, sex hygiene knowledge and sexually transmitted diseases (STDs), etc.</p><p><b>RESULTS</b>39.33% (459/1167) students thought they had accepted formal sex education; 49.19% (574/1167) students chose lecture as the favorite way; 41.90% (489/1167) students thought the reproductive health education would not increase the incidence of premarital sexual behavior; 64.10% (748/1167) students thought it was necessary to give college students reproductive health instructions and knowledge. On the other hand, we found 45.05% (250/555) boys and 27.45% (168/612) girls could accept the premarital sexual behaviors; 35.32% (196/555) boys and 39.38% (241/612) girls could accept premarital pregnancy; 44.39% (518/1167) students could calculate the safe period accurately; 71.38% (833/1167) students knew emergency contraception measures; 57.67% (673/1167) students knew how to and when to use condoms; To our surprise, only 2.19% (12/549) non-medical students and 12.14% (75/618) medical students knew exactly the whole 4 sexually transmitted diseases that needed to be essentially prevented in China.</p><p><b>CONCLUSION</b>At present, the reproductive health condition of college students in Shanghai is sophisticated. Much more education on sex safety, self-protection skills and acquired immunodeficiency syndrome (AIDS) knowledge, etc should be reinforced.</p>

Expression and its significance of TRAIL and its receptors in cells of patients with acute myeloid leukemia / 中国实验血液学杂志

This study was aimed to detect the expression of TNF related apoptosis-inducing ligand (TRAIL) and its receptors on acute myeloid leukemic (AML) cells, and explore its possible role in leukemia therapy. RT-PCR and flow cytometry were used to detect the expression of TRAIL and its receptors on AML cells of 39 cases (patient group), AML cells of 18 cases with complete remission (CR group) and BMMNC or PBMNC of 21 normal persons (control group). The results showed that (1) TRAIL, DR4 and DR5 were highly expressed in both patient group and CR group, while the DcR1 and DcR2 were poorly expressed. (2) The level of DR5 expression in CR group was higher than that in patient group. (3) The level of DR5 was higher than DR4 in both patient group and CR group. (4) TRAIL and its receptors were expressed similarly in different subtypes of AML. In conclusion, there are differences between the expressions of TRAIL and its receptors in AML cells. DR5 may play an important role in TRAIL-inducing apoptosis of AML cells.

Apoptosis of acute myeloid leukemic cells induced by human recombinant soluble tumor necrosis factor-related apoptosis inducing ligand (TRAIL) / 中国实验血液学杂志

In order to investigate the effect of tumor necrosis factor-related apoptosis inducing ligand (TRAIL) on acute myeloid leukemic (AML) cells, BMMNC or PBMNC of 39 cases of AML (patient group) and 21 normal persons (control group) were isolated; the cytotoxic effects of TRAIL at various concentration on the cells were evaluated by MMT, apoptosis were detected by flow cytometry. The results showed that TRAIL of various concentration could kill AML cells in various degrees, but had no killing effect on normal cells. It is concluded that TRAIL can kill AML cells through inducing apoptosis, and TRAIL has a promising prospect in clinical use.

Expansion of marrow hematopoietic progenitor cells ex vivo supported by culture system from mouse marrow embryonic fibroblasts and leukemia inhibitory factor / 中国实验血液学杂志

The objective of this study was to explore the effect of culture system from embryonic fibroblasts and leukemia inhibitory factor (LIF) on expansion of mouse bone marrow hematopoietic progenitor cells ex vivo, and to observe its effect on the expression of homing-related cell adhesion molecules among VLA-4 (CD49e), VLA-5 (CD49e), LFA-1 (CD11a), HCAM (CD44) and L-selectin (CD62L). The culture system from the mouse embryonic fibroblasts inactivatd by mitomycin C and contained LIF was used to culture with mouse BMMNC for 7 days. The total number of BMMNC, CFC, Sca-1(+) cells, cell apoptosis rate and the expression of above cell adhesion molecules were counted. The results showed that culture system consisted of embryonic fibroblasts and LIF significantly enhanced the total number of BMMNC, CFC, Sca-1(+) cells, suppressed cell apoptosis (P < 0.05). In control without MEF and LIF, the total number of BMMNC was reduced remarkedly, CFC and Sca-1(+) cells were completely dead, the majority of cells produced apoptosis (P < 0.01). The expression of CD49d, Cd44 and CD61L on Sca-1(+) cells were similar to that befor expression (P < 0.05), but the expression of CD49e and CD11a on Sca-1(+) cells were remarkably increased (P < 0.05). It is concluded that culture system from embryonic fibroblasts and LIF can only significantly expand mouse bone marrow hematopoietic progenitor cells ex vivo, but the expanded hematopoietic progenitor may well sustain the expression of homing-related adhesion molecules. The homing functions of these expanded hematopoietic progenitors kept no change.