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T cells in grafts serve an important role in the pathogenesis of graft versus host disease (GVHD) and immune recovery during HLA matched allogeneic stem cell transplantation. However, the role of T cells in the haploidentical peripheral blood stem cell transplantation (Haplo-PBSCT) is yet to be determined. In the present study, the role of CD3+ T cells in grafts and impact on hematopoietic and immune recovery, cytomegalovirus (CMV) reactivation, GVHD, relapse, progress free survival and overall survival (OS) were evaluated and analyzed. A total of 30 patients who underwent haplo-PBSCT were included in the present study. CD3+ T cells accounted for a median of 23.1% (range 8-47.4%) with a median dose of 299.7x106/kg (range 104-623.4). Patients were divided into two groups according to the CD3+ T cell count: Above the median (high T cell group) and below the median CD3+ T cell (low T cell group). No significant difference was identified between neutrophil and platelet recovery time between two groups (P>0.05). The mean lymphocyte recovery time of high T cell group and low T cell group were 107.07 days (95% CI 79.88-134.25), and 50.4 days (95% CI 41.42-59.38), respectively. The lymphocyte recovery time of high T cell group was higher that of low T cell group (P 0.05). The cumulative incidence of grade II or above acute GVHD was higher in the high T groups compared with low T groups (P

Additional file 1. Standard Protocol Items: Recommendations for Interventional Trials (SPIRIT) 2013 checklist.

T cells in grafts serve an important role in the pathogenesis of graft versus host disease (GVHD) and immune recovery during HLA matched allogeneic stem cell transplantation. However, the role of T cells in the haploidentical peripheral blood stem cell transplantation (Haplo-PBSCT) is yet to be determined. In the present study, the role of CD3(+) T cells in grafts and impact on hematopoietic and immune recovery, cytomegalovirus (CMV) reactivation, GVHD, relapse, progress free survival and overall survival (OS) were evaluated and analyzed. A total of 30 patients who underwent haplo-PBSCT were included in the present study. CD3(+) T cells accounted for a median of 23.1% (range 8-47.4%) with a median dose of 299.7x10(6)/kg (range 104-623.4). Patients were divided into two groups according to the CD3(+) T cell count: Above the median (high T cell group) and below the median CD3(+) T cell (low T cell group). No significant difference was identified between neutrophil and platelet recovery time between two groups (P>0.05). The mean lymphocyte recovery time of high T cell group and low T cell group were 107.07 days (95% CI 79.88-134.25), and 50.4 days (95% CI 41.42-59.38), respectively. The lymphocyte recovery time of high T cell group was higher that of low T cell group (P0.05). The cumulative incidence of grade II or above acute GVHD was higher in the high T groups compared with low T groups (P

Monitoring minimal residue disease (MRD) is an effective approach to evaluate the response to chemotherapy, and it is used to select the ideal therapeutic strategy and to predict the recurrence during therapy for hematological disorders. The Wilm's tumor 1 (WT1) gene, which is highly expressed in >80% of patients with acute myeloid leukemia (AML) and its increased expression level may cause poor clinical outcomes, is a potential MRD marker of hematological neoplasms. In the present study, the expression levels of WT1 and other molecular markers were retrospectively analyzed by reverse transcription‑quantitative PCR in 195 patients with AML. The expression level of WT1 was significantly lower in patients with remission compared with patients with early‑stage and recurrent AML. Moreover, WT1 expression was significantly decreased in patients with RUNX family transcription factor 1‑RUNX1 translocation partner 1 fusion, but higher in patients with promyelocytic leukemia‑retinoic acid receptor α fusion. WT1 expression was significantly reduced during remission. In patients with AML who underwent allogeneic hematopoietic stem cell transplantation (allo‑HSCT), the mortality rate 2 years after allo‑HSCT was significantly lower in patients with low expression level of WT1 compared with subjects presenting high expression level of WT1. Collectively, the upregulation of the expression level of WT1 in combination with the identification of other genetic abnormalities may be used as MRD markers of hematological neoplasms.

Standard Protocol Items: Recommendations for Interventional Trials (SPIRIT) 2013 Checklist. (DOC 123 kb)

Background The treatment of acute promyelocytic leukemia (APL) has been revolutionized in the past two decades by the advent of all-trans retinoic acid (ATRA) and arsenic trioxide (ATO). It suggests that non-high-risk APL patients can be cured without chemotherapy. However, ATRA plus chemotherapy is still the standard therapy for the high-risk patients. Central nervous system (CNS) relapse remains a significant cause of treatment failure in high-risk patients. However, increasing the ATO concentration in cerebrospinal fluid (CSF) may reduce CNS relapse in high-risk patients. Mannitol can allow ATO to penetrate the blood-brain barrier (BBB) and reach therapeutically effective levels in the CSF. It is used for the treatment of CNS relapse in patients APL. We compare ATRA-ATO with ATRA-ATO plus chemotherapy in both high-risk and non-high-risk patients with APL. Methods This study was designed as a multicenter randomized controlled trial. Patients with APL were randomly assigned into two groups: the ATRA-ATO group (experimental group) and the ATRA-ATO plus chemotherapy group (control group). The experimental group receives therapy with ATRA-ATO for induction, consolidation and maintenance therapy. In the high-risk patients, mannitol will be used with ATO in the consolidation and maintenance therapy. Hydroxyurea will be used in patients who developed leukocytosis in the induction therapy. The control group receives therapy with ATRA-ATO plus chemotherapy for induction and consolidation therapy. Discussion In this study, a randomized clinical trial design is described. It aims to compare the efficacy of ATRA-ATO versus ATRA-ATO plus chemotherapy in all-risk patients with APL. Trial registration Chinese Clinical Trials Registry, ID: ChiCTR-IPR-15006821. Registered on 27 July 2015. Electronic supplementary material The online version of this article (10.1186/s13063-018-2812-3) contains supplementary material, which is available to authorized users.

Contents: Preface. Chinese Children: Introduction to Chinese children, Yunxiao Sun Introduction to Chinese children: a commentary, Yan Wang Chinese children's physical and mental development, Hongyan Sun Chinese children's physical and mental development: a commentary, Ginny Qin Zhan Chinese children's leisure, Weidong Chen Chinese children's leisure: a commentary, Vicky Chiu-wan Tam Chinese children's consumption, Xia Zhao Chinese children's consumption: a commentary, Bryan Tilt. Chinese Youth: Introduction to Chinese youth, Jieying Xi Introduction to Chinese youth: a commentary, Yan Xia Chinese youth's employment, Guoqi An Chinese youth in rural areas, Hua Zhang Chinese youth in rural areas: a commentary, I. Joyce Chang Chinese college students, Junyan Liu Chinese college students: a commentary, Baomei Zhao and Raymond E. Forgue. Chinese Children and Youth: Popular culture among Chinese youth, Changzheng Yang Popular culture among Chinese youth: a commentary, Shelley Hong Xu Only children in China, Yunxiao Sun and Xia Zhao Only children in China: a commentary, Li Huang Internet use among Chinese youth, Wei Bu Internet use among Chinese youth: a commentary, Xin-an Lu Juvenile delinquency in China, Ying Guan Juvenile delinquency in China: a commentary, Hong Lu Index.

Different countries and fields of study differ in determining the age group of youth. Based on the census result from the National Bureau of Statistics and the situation of Chinese youth, the age group of youth is defined as those from 15 to 29 years old. Therefore, Chinese youth described in this chapter refers to the age group of 15-29 living in the Chinese mainland, not including youth from Hong Kong, Macao Special Administrative Regions and Taiwan Province.

In order to investigate the effect and mechanisms of interferon (IFN)-γ in combination with all-trans-retinoic acid (ATRA) on NB4 cells [ATRA-sensitive acute promyelocytic leukemia (APL) cell line] and NB4-R1 cells (ATRA-resistant APL cell line) and to search for a novel approach to solve the problem of ATRA resistance in APL, we initially treated NB4 and NB4-R1 cells with IFN-γ, ATRA and IFN-γ in combination with ATRA, respectively. The cell proliferation was then tested by MTT assay, and the cell differentiation was tested through light microscopy, by NBT test and flow cytometry (FCM). The expression of promyelocytic leukemia (PML) protein was observed by indirect immune fluorescent test. Results showed that ATRA inhibited the growth of NB4 cells, however, it could not inhibit the growth of NB4-R1 cells. IFN-γ inhibited the growth of both NB4 and NB4-R1 cells. Meanwhile, the growth inhibition effect of IFN-γ in combination with ATRA on both NB4 and NB4-R1 cells was significantly stronger than that of any single drug treatment. The results of the NBT reduction test and CD11b antigen detection by FCM indicated that IFN-γ induces the differentiation of NB4 and NB4-R1 cells to some extent. Moreover, the maturation degree of both NB4 and NB4-R1 cells induced by IFN-γ in combination with ATRA was more significant than that of IFN-γ or ATRA alone. After treatment with IFN-γ, the number of fluorescent particles in NB4 and NB4-R1 cell nuclei was higher than those in the control group, which indicated that IFN-γ may induce the expression of PML protein. Together, IFN-γ augments the proliferation inhibition effect of ATRA on NB4 and NB4-R1 cells through enhancing the expression of PML protein. IFN-γ in combination with ATRA not only strengthens the induction differentiation effect of ATRA on NB4 cells, but also can partially induce the maturation of NB4-R1 cells with ATRA resistance.

Rationale: Papillary thyroid carcinoma (PTC) is the most common endocrine malignancy. However, the simultaneous occurrence of PTC and Hodgkin Lymphoma (HL) was rarely reported. Patient concerns: We present a case of simultaneous BRAFV600E-positive PTC and HL in a 17-year-old female. Diagnosis: She was referred to our clinic with a painless lump in her left neck. A highly suspicious thyroid nodule and multiple enlarged lymph nodes in the neck were found by ultrasonography examination. The suspicious nodule was diagnosed as PTC by fine needle aspiration cytology. Interventions: A total thyroidectomy with bilateral lymph node dissection was performed and the microscopic examination revealed a 2-cm PTC with BRAFV600E mutation and HL (mixed cellularity) in the bilateral lymph nodes. PTC was postoperatively considered as T1bN0M0. Levothyroxine (125 μg/d) was administered to the patient for thyrotropin suppression therapy. Then the patient was referred to the Department of Hematology to receive 4 cycles of ABVD followed by 30 Gy involved-site radiotherapy and radioactive iodine (RAI) therapy for thyroid cancer. Outcomes: After two cycles of ABVD, multiple enlarged lymph nodes showed a significant response to the chemotherapy in the patient. Lessons: Simultaneous HL and BRAFV600E-positive PTC is extremely rare. Biopsy of the suspicious lymph nodes should be performed to confirm malignancy metastasizing from PTC or other lesions. Similarly, in HL patients with suspicious thyroid nodule, ultrasound-guided fine needle aspiration of thyroid nodule should be performed to exclude thyroid malignancy.

Objective To study the effect of natural killer (NK) cells on graft-versus-host disease (GVHD) after H-2 haploidentical bone marrow transplantation (BMT) in mice. Methods Murine model of H-2 haploidentical BMT was established by using Balb/c (H-2d) mouse as recipient, and Balb/c (H-2d)xC57BL/6 (H-2b)(H-2d/b) mouse as donor. Lethally irradiated Balb/c (H-2d) mice were transplanted with the bone marrow cells from Balb/c(H-2d)xC57BL/6(H-2b)(H-2d/b) mice containing donor spleen cells and/or NK cells. GVHD and survival rates were studied by observation of clinical manifestations and pathological changes. Results In the group of bone marrow +spleen cells, GVHD was induced in 90% mice; but in the group plus with low amount of NK cells, GVHD was induced in 20% mice; and in the group transplanted with high amount of NK cells, GVHD was induced only in 10% mice. Compared to the group transplanted only with BM plus spleen cells, the incidences of GVHD in the latter two groups decreased significantly (P Conclusion In mouse H-2 haploidentical BMT, alloreactive NK cells can reduce the incidence of GVHD and increase the survival rate.

Arsenic trioxide (ATO) combined with dexamethasone, melphalan or other cytostatic agents had been used to treat refractory or relapsed multiple myeloma (MM) patients. We assessed the safety and efficacy of ATO combined with vindesine/cyclophosphamide/melphalan/prednisone (VCMP) or vindesine/doxorubicin/dexamethsone (VAD) chemotherapy for MM patients who failed more than two different prior regimens. All patients received ATO (0.25 mg/kg day) for 10 days/cycle combined with VCMP or VAD in 30-day cycles. Vindesine (1.4 mg/m(2)) was given intravenously on day 1, cyclophosphamide (400 mg/m(2) day) was given intravenously on days 2, 4, 6, 8, 10, melphalan (6 mg/day) and prednisone (1 mg/kg day) were given orally day 1 to day 10 for VCMP regimen. VAD regimen consisted of vindesine 1 mg/day and doxorubicin 10 mg/day intravenously drip for 4 days with oral dexamethasone 40 mg/day for days 1-4, 9-12, 17-20. Patients who completed at least one cycle were evaluated for response to treatment. Objective responses occurred in 35 of 63 (56 %) patients, including seven complete, 14 partial and 14 minor responses. Median progression-free survival and overall survival were 6 and 23 months respectively. 12 patients had elevated serum creatinine levels (SCr) at baseline, and 9 of 12 (75 %) showed decreased SCr levels during treatment. Frequent Grade 3/4 non-hematological adverse events included arrhythmia, hypertension, fatigue and neuropathy. These results indicate that ATO combined with VCMP or VAD was effective and well tolerated as a new therapeutic option for patients with relapsed or refractory MM.

To explore optimum conditions for establishing a two‑dimensional gel electrophoresis (2-DE) map of the human acute promyelocytic leukemia (APL) cell line NB4 and to analyze its protein profiles, we extracted total proteins from NB4 cells using cell disruption, liquid nitrogen freeze-thawing and fracturing by ultrasound, and quantified the extracted protein samples using Bradford's method. 2-DE was applied to separate the proteins, which were silver-stained in the gel. Well‑separated protein spots were selected from the gel using the ImageMaster™ 2D Platinum analysis system. Moreover, the effects of various protein sample sizes (140, 160 and 180 µg) on the 2-DE maps of the NB4 cells were determined and compared. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS), peptide mass fingerprinting (PMF) and database searching were used to identify the proteins. When the quantity of loading proteins was 160 µg, clear, well-resolved, reproducible 2-DE proteomic profiles of the NB4 cells were obtained. The average number of protein spots in 3 gels was 1160±51 with an average matching rate of 81%. A total of 10 proteins were identified by mass spectrometry and database queries, certain proteins were products of oncogenes and others were involved in cell cycle regulation and signal transduction. In summary, 2-DE profiles of the proteome of NB4 cells were established and certain proteins were identified by MALDI-TOF-MS and PMF which lay the foundation of further proteomic research of NB4 cells. These data should be useful for establishing a human APL proteome database.

Although 90% patients with untreated acute promyelocytic leukemia(APL) obtain complete remission because of the usage of all-trans retinoic acid(ATRA), patients with ATRA-resistance are increased gradually. ATRA-resistance has become one of the main causes which affect the long-term therapeutic efficacy of APL. The mechanisms of ATRA-resistance are complex, which probably involve the metabolism of ATRA, abnormal expression of cellular retinoic acid binding protein(CRABP) and P-glycoprotein(P-gp), mutation of RARα and aberration translocation of APL. However, in these previous researches, it was one or a few proteins but not the entirety proteins that were emphasized on the mechanisms of ATRA-resistance. Comparative proteomics can analyze the entire protein expression in cells in whole and has the superiority in screening the drug-resistance proteins differentially expressed. In order to investigate the mechanisms of ATRA-resistance in APL in whole, we compared and analyzed the protein expression profiles between MR2 cells(APL cell line with ATRA-resistance) and NB4 cells(APL cell line with ATRA-sensitiveness) by comparative proteomics. After the total proteins of MR2 cells and NB4 cells were extracted respectively, they were separated by two-dimensional electrophoresis(2-DE). The differences in proteome profile between MR2 cells and NB4 cells analyzed by ImageMaster™ 2D Platinum software. The average protein spots in 2-DE maps of MR2 and NB4 cells were 1160±51 and 1068±33 respectively. 8 protein spots were selected to be identified by Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS), in which the quantity of the protein differentially expressed was more than two times(≥2 or ≤0.5) between MR2 and NB4 cells’ 2-DE map. They were all successfully identified and their definite information was obtained. Among them, 6 proteins were probably involved in the mechanisms of ATRA-resistance in APL and they were Cofilin-1, Elongation factor 1-beta (EF-1β), Tropomyosin isoform(TM), High mobility group protein B1(HMGB1), Ran-specific GTPase-activating protein (RanGAP1) and Galectin-1. Moreover, so far there was no related report on the roles of HMGB1, RanGAP1 and Galectin-1 in the mechanisms of ATRA-resistance in APL. These differential proteins identified provide the new clues for us to further elucidate the mechanisms of ATRA-resistance from multiple factor.

More than 90% patients with untreated acute promyelocytic leukemia(APL) obtain complete remission clinically by using All-trans retinoic acid(ATRA), a strong differentiation inducer. However, the rapid development of ATRA-resistance brings a new problem to the treatment of APL. Interferon(IFN), as an important cytokine, has broad biological activities. It can not only inhibit the growth of tumor cells, but also reverse the drug-resistance of chemotherapy. As proved by some research, the mechanisms of ATRA-resistance are probably related to lacking some important proteins which synthesized by Interferon-γ (IFN-γ). In order to explore a new way to solve the problem of ATRA-resistance in APL, we investigate the effect and mechanisms of IFN-γ in combination with ATRA on the proliferation/differentiation of NB4 cells(APL cell line with ATRA-sensitiveness) and MR2 cells(APL cell line with ATRA-resistance) respectively. ATRA, IFN-γ and IFN-γ in combination with ATRA were incubated with NB4 and MR2 cells respectively. The cell proliferation was tested by MTT assay, the cell differentiation was tested through light microscope, by NBT reduction test and flow cytometry(FCM). The results showed that ATRA could inhibit the growth of NB4 cells significantly (P0.05). IFN-γ could inhibit the growth of both NB4 cells and MR2 cells slightly (P0.05). Although IFN-γ alone had no effect on the differentiation of either NB4 or MR2 cells (P>0.05), it could augment the differentiation of NB4 cells induced by ATRA (P

Allogeneic haemopoietic stem-cell transplantation (HSCT) is a definitive cure for many malignant blood diseases. Ideal donors for transplantation are those with completely matched human leukocyte antigen (HLA), kin-relative or no-kin-relative. However only about 25% to 30% people could find HLA matched related-donor and there is only 1/100,000 opportunity to find a matched HLA in unrelated group. Recent years HLA haploidentical transplantation play more and more important role in across the histocompatibility barrier and make about 90% patients possible to get transplantation. However GVHD is still main problem during the transplantation. Recent studies show that natural killer (NK) cells could specially attack recipient antigen-presenting cells (APCs), shown to be responsible for decreasing and improving GVHD. To explore the effect of NK cell on GVHD after H-2 haploidentical bone marrow transplantation(BMT) in mice, we purified NK cells by positive selection with CD49b (DX5) MicroBeads (Miltenyi Biotec product) from donor mice as ingredient in the conditioning regimens, and observed the influence of donor NK cells on GVHD and evaluated the potential role of donor NK cells in protecting against GVHD. Murine model of H-2 haploidentical BMT was established by using Balb/c(H-2d) mouse as recipient, and Balb/c (H-2d)×C57BL/6 (H-2b)(H-2d/b) mouse as donor. Lethally irradiated Balb/c(H-2d) mice were transplanted with Balb/c(H-2d)×C57BL/6(H-2b)(H-2d/b) bone marrow containing donor peripheral T cells and/or NK cells. GVHD and survival rate were studied by observing clinical manifestations and pathological changes. In the group with bone marrow plus T cells, GVHD was induced in 90% mice; but in the group plus with low concentration of NK cells, GVHD was induced in 20% mice; and in the group transplanted with high concentration of NK cells, GVHD was induced only in 10% mice. Compared with the group transplanted only with T cells, the incidences of GVHD in the latter two groups decreased obviously (P

Although all-trans retinoic acid(ATRA) provides complete remission in 90% patients with untreated acute promyelocytic leukemia(APL), it becomes ineffective to quite a few APL patients who have received ATRA before when their disease relapsed and used ATRA again. Arsenic trioxide(ATO) can make APL patients with ATRA-resistance obtain complete remission again by inducing APL cells apoptosis. However, the molecular mechanisms of apoptosis in ATRA-resistance APL cells induced by ATO remain unclear. For this reason, we take the apoptotic MR2 cells (APL cell line with ATRA-resistance) induced by ATO as a model, to screen and identify the proteins related with ATO-induced apoptosis by comparative proteomics. After MR2 cells were dyed with annexin V and PI staining, the percentage of the apoptotic MR2 cells induced by 1.0μmol/L ATO for 0h, 6h, 12h, 24h and 48h respectively was detected by Flow cytometry. The results showed that the majority of the apoptotic cells were in the earlier and later stage of apoptosis respectively, when MR2 cells were treated with ATO for 24 and 48 hours in sequence. The total proteins of MR2 cells of the control group, the earlier stages apoptosis group and the later stages apoptosis group were separated by two-dimensional electrophoresis(2-DE) respectively. Then, the differences in proteome profile among three groups were analyzed by ImageMaster™ 2D Platinum software. 14 protein pots were selected to be identified by Matrix-assisted laser desorption/ionization time of flight-mass spectrometry(MALDI-TOF-MS), in which the quantity of the protein differentially expressed was more than two times(≥2 or ≤0.5) among MR2-0h, MR2-24h and MR2-48h cells’ 2-DE map. However, only 11 proteins were successfully identified and their definite information was obtained. Among them, there were 8 proteins that were all probably involved in the mechanisms of apoptosis in MR2 cells and they were Calreticulin(CRT), Heat shock 70 kDa protein(HSP70), High mobility group protein B1(HMGB1), Ran-specific GTPase-activating protein(RanGAP1), Elongation factor 1-beta(EF-1β), Beta-tubulin, Cofilin-1, and Prolyl 4-hydroxylase(P4H) respectively. CRT was probably related with the early stage of apoptosis in MR2 cells, while RanGAP1 and HSP70 might related with the late stage of apoptosis in MR2 cells. Moreover, so far there was no related report on the roles of CRT, HMGB1, RanGAP1, cofilin-1 and beta-tubulin in the mechanisms of APL cells apoptosis. These differential proteins identified provide the new clues for further researching the molecular mechanisms of apoptosis in the ATRA-resistance APL cells induced by ATO.