Dehydroabietic acid chemosensitizes drug-resistant acute lymphoblastic leukemia cells by downregulating survivin expression

Objective: To explore the mechanism of drug resistance in acute lymphoblastic leukemia and the anti-tumor effect of combination therapy of dehydroabietic acid and vincristine against acute lymphoblastic leukemia cells. Methods: Acute lymphoblastic leukemia cells REH and CCRF- CEM were employed to detect the anti-tumor effect of vincristine and doxorubicin on proliferation and apoptosis using EdU assay, human active caspase-3 Quantikine ELISA kit, and flow cytometry. Vincristine-resistant REH cells (REH-R), survivin knockdown and overexpressing REH cells were established to verify the role of survivin in drug resistance. Additionally, in vitro and in vivo assays were performed to determine the effect of dehydroabietic acid on the cytotoxicity of vincristine. Results: Vincristine and doxorubicin markedly suppressed proliferation and induced apoptosis of REH and CCRF-CEM cells. Survivin expression was upregulated in REH-R cells compared with REH cells. Knockdown of survivin expression obviously restored the sensitivity of REH-R cells to vincristine. Akt phosphorylation was also increased in REH-R cells compared to REH cells. In addition, LY294002, a PI3k/Akt pathway blocker, inhibited survivin expression and enhanced cytotoxicity of vincristine to REH-R cells. Dehydroabietic acid effectively reduced survivin expression in REH-R cells, thereby enhancing the therapeutic effect of vincristine on drug-resistant cells. Survivin overexpression markedly reduced the effect of dehydroabietic acid on enhancing the anti-proliferation and inducing apoptosis effect of vincristine. Moreover, the combination of dehydroabietic acid with vincristine significantly extended the survival rate in a mouse xenograft model of acute lymphoblastic leukemia, compared with vincristine treatment alone. Conclusions: Dehydroabietic acid may be used as a potential candidate for the treatment of acute lymphoblastic leukemia in combination with vincristine.

Effects of Proton Pump Inhibitors on the Gastrointestinal Microbiota in Gastroesophageal Reflux Disease / 基因组蛋白质组与生物信息学报·英文版

Proton pump inhibitors (PPIs) are commonly used to lessen symptoms in patients with gastroesophageal reflux disease (GERD). However, the effects of PPI therapy on the gastrointestinal microbiota in GERD patients remain unclear. We examined the association between the PPI usage and the microbiota present in gastric mucosal and fecal samples from GERD patients and healthy controls (HCs) using 16S rRNA gene sequencing. GERD patients taking PPIs were further divided into short-term and long-term PPI user groups. We showed that PPI administration lowered the relative bacterial diversity of the gastric microbiota in GERD patients. Compared to the non-PPI-user and HC groups, higher abundances of Planococcaceae, Oxalobacteraceae, and Sphingomonadaceae were found in the gastric microbiota from the PPI-user group. In addition, the Methylophilus genus was more highly abundant in the long-term PPI user group than in the short-term PPI-user group. Despite the absence of differences in alpha diversity, there were significant differences in the fecal bacterial composition of between GERD patients taking PPIs and those not taking PPIs. There was a higher abundance of Streptococcaceae, Veillonellaceae, Acidaminococcaceae, Micrococcaceae, and Flavobacteriaceae present in the fecal microbiota from the PPI-user group than those from the non-PPI-user and HC groups. Additionally, a significantly higher abundance of Ruminococcus was found in GERD patients on long-term PPI medication than that on short-term PPI medication. Our study indicates that PPI administration in patients with GERD has a significant effect on the abundance and structure of the gastric mucosal microbiota but only on the composition of the fecal microbiota.

ARDS severity identification based on noninvasive parameter SpO2/FiO2 / 医疗卫生装备

Objective To explore the relationship between SpO2/FiO2(S/F) and PaO2/FiO2(P/F) so as to determine the possibility of ARDS severity identification based on noninvasive parameters. Methods The physiological parameters of corresponding patients were acquired from Medical Information Mart for Intensive Care (MIMIC-Ⅲ),and then divided into a training set and a test set randomly.In the training set the linear relationship between lg(S/F)and lg(P/F)was established with generalized linear regression model,and a log linear regression model was formed with the optimal regression equation;the linear relationship between lg(S/F)and lg(P/F)was compared with that between S/F and P/F.In the test set,the two models were compared on the identification of ARDS in case P/F values were 100(mild ARDS),200(moderate ARDS)and 300(severe ARDS)respectively.Results In the training set(n=61 634)the linear relationship between lg(S/F)and lg(P/F)was deduced as lg(S/F)=1.277+0.437×lg(P/F) (r=0.66,P<0.000 1),and the S/F thresholds in case P/F values were 100,200 and 300 respectively were 131,201 and 271.In the test set (n=26 758)the identification effect was verified with the acquired S/F thresholds,which proved better than that of traditional regression model.Conclusion Noninvasive parameter SpO2/FiO2can replace PaO2/FiO2for the auxiliary diagnosis of ARDS in case the result of blood gas analysis is absent.

Management of medical equipment in hospital ship / 医疗卫生装备

Objective To explore the medical equipment management to facilitate the medical support in the hospital ship.Methods The influences and the ship-mounted equipment on medical equipment were analyzed,the requirements of sea conditions for medical equipment management were explored,the hidden risks and present situation of medical equipment were introduced,and some countermeasures were put forward accordingly.Results The problems of equipment management were analyzed in hospital ship,and some countermeasures were brought out including bringing into better balance the relations between various sectors,enhancing personnel,introducing professional talents,trying for the support from the director and etc.Conclusion The safety of hospital medical equipment is of great significance to the hospital ship itself and the patient.Only to strengthen medical equipment management while focusing on the qualified personnel contributes tobetter fulfilling its mission.

Pharmacokinetics of antithymocyte globulin in the treatment of childhood aplastic anemia / 中华实用儿科临床杂志

Objective To investigate the pharmacokinetics of antithymocyte globulin(ATG) in the treatment of childhood aplastic anemia(AA).Methods Six cases of of childhood AA treated with rabbit ATG (R-ATG,Genzyme company,USA and Fresenius company,Germany)were in the study.Enzyme-linked immunosorbent assay (ELISA) was used to detect the concentrations of ATG in plasma during the treatment period of the first day to the 5th day,and during the 7th,21th,35th,60th and 90th after the treatment finished respectively(all 12 time nodes).The pharmacokinetic data was detect with the Practical Pharmacokinetics-program(3P97).Results The peak levels of ATG concentrations in plasma(30-32 mg/L)were reached at the third to forth day during the treatment period,and then gradually descended to about half of the peak level in the 21st day(16-18 mg/L),and the microcomponent of ATG in plasma could be detected in the 90th day after the therapy.There were not evident difference of plasma concentrations in every time nodes,the peak levels,and pharmacokinetic data including the drug half-life,area under the plasma concentration-time curve (AUC) between the 2 forms of R-ATG (Genzyme company,USA and Fresenius company,Germany).The therapy effective rate in the 6 cases AA of this group was 66.7％ in the followed-up for mean 12 months.Conclusions Successfully set up the techniques to detect the pharmacokinetics of ATG in the treatment of childhood AA,will help to research the relationship between the pharmacokinetics with the side effects and outcomes of ATG in the treatment of AA.

Overview of clinical occurrence of primary immunodeficiency disorders in children / 中华儿科杂志

<p><b>OBJECTIVE</b>More than one hundred primary immunodeficiency disorders have been discovered so far. But the incidence of these disorders in our country is still not clear, so we analyzed the clinical data of 93 children with primary immunodeficiency disorders seen in our hospital in recent 30 years to understand the occurrence of primary immunodeficiency disorders in children, to promote the clinicians to become familiar with these disorders, to improve the nationwide registry system and to establish the basis for the treatment and prevention in future.</p><p><b>METHODS</b>To analyze the constituent ratio of the 93 children with primary immunodeficiency disorders seen in our hospital from 1974 to 2003, diagnostic and classification criteria were set by taking the proposal by International Union of Immunological Societies (IUIS) PID classification committee in 2003 into account. All the data were analyzed retrospectively.</p><p><b>RESULTS</b>In the 93 children with primary immunodeficiency disorders, antibody deficiencies were the most frequent (39.8%) finding, followed by combined immunodeficiency, combined T- and B-cell disorders (22.6%), and T lymphocytic deficiencies alone (14.0%). Immunodeficiency with other major defects accounted for 12.9%, phagocytic disorders 9.7%, and complement deficiencies 1.1%. Thus, there seemed to be a tendency that the incidence increased with time. The incidence of these disorders has increased significantly as shown by 50 diagnosed cases in children with these disorders since 1996. Sixteen children died, with the highest mortality occurred with combined immunodeficiency. Seven children developed bronchiectasis. Two children suffered from persistent diarrhea while one of the two was complicated with persistent intestinal fistula. One child developed juvenile rheumatoid arthritis, another one with granulocytopenia and iridocyclitis, and the other with allergic purpura. The boys: girls ratio for all disorders was 3:1. The age of onset ranged from 10 days to 37 years of age.</p><p><b>CONCLUSIONS</b>There are vast variety of primary immunodeficiency disorders in our area and antibody deficiency is the most common abnormality. Combined immunodeficiency has early onset age and high mortality rate. With the great improvement of the diagnostic techniques, these disorders have become a group of important disorders and all the clinicians should pay great attention to these disorders.</p>

Prognostic significance of lymphocyte function associated anti-gen-3 (CD58) in childhood B cell-acute lymphocytic leukemia / 中国实验血液学杂志

This study was aimed to investigate the value of CD58 in evaluation of early therapeutic effect on childhood B-ALL. The expression features of CD58 in 135 cases of childhood B-ALL were analyzed by four-color flow cytometry; MRD detection protocol for B-ALL using CD58/CD10/CD34/CD19 combination was established; the correlation between the expression features of CD58 and MRD detection was analyzed for the early therapeutic response in childhood B-ALL. The results showed that the mean value of CD58 MFI in 135 cases of B-ALL was 113.08 +/- 63.33, which was significantly higher than that in 15 cases of normal bone marrow controls (14.68 +/- 5.26, P < 0.01). In addition, CD58 was over expressed in 51.9% (70/135) of B-ALL patients, indicating that CD58 could be an effective marker in MRD detection. The CD58/CD10/CD34/CD19 was the second most effective combination next to TdT/CD10/CD34/CD19 in B-ALL MRD detection with flow cytometry. Meanwhile, the positive rate of MRD detection by flow cytometry was significantly lower in CD58 over expression group (P < 0.05). It is concluded that CD58 may be used as an indicator for detection of MRD in B-ALL patients, which would enrich the combination of MRD detection. The CD58 over expression may be considered as a marker of a favorable prognosis in childhood B-ALL.

Expression of retinoblastoma protein in child acute leukemia cells and its clinical significance / 中国实验血液学杂志

The research was aimed to detect the expression levels of retinoblastoma protein (pRb) in child acute leukemia cells, and to explore its possible association with leukemia cells cycle, the risk of disease, minimal residual disease (MRD) monitoring and prognosis of B-ALL. Flow cytometry (FCM) was used to detect the expression of pRb in 89 cases of acute leukemia (including 25 AML, 10 T-ALL and 54 B-ALL) and bone marrows from 7 normal children (control group). Meanwhile the cell cycle in some cases was analyzed. The results showed that (1) the FCM could accurately detect the expression of pRb in acute leukemia cells; (2) the high level of pRb expression was frequent in all types of child acute leukemias. In the same case, the expression of pRb was significantly increased in leukemia cells when compared with non-leukemia cells. And no detectable pRb protein was found in partial cases of acute leukemia; (3) there was a close relation between expression of pRb and the cell cycle of leukemia cells, the number of G(1) phase cells in pRb positive case of B-ALL was more than that in pRb negative case (92% vs 77%); (4) in B-ALL, the level of pRb expression in MRD positive group was significantly lower than that in MRD negative group (P < 0.05), but pRb expression was stable in non-leukemia cells during therapy; (5) pRb expression was related to the early response to therapy in B-ALL, the expression of pRb was significantly increased in sensitive group when compared with insensitive group (P < 0.05). It is concluded that high level or absence of pRb expression can be found in child acute leukemia cells. The expression of pRb is positively related to cell cycle of leukemia cells, MRD monitoring and the early response to therapy. In short, the detection of pRb expression level can guide the therapy and the evaluation of prognosis in B-ALL.

Analysis of therapeutic effectiveness in 158 childhood acute lymphoblastic leukemia patients treated with ALL-XH-99 protocol / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyse the treatment outcome of 158 childhood acute lymphoblastic leukemia (ALL) patients, and explore how to improve the event-free survival (EFS) rate in ALL.</p><p><b>METHOD</b>All of the patients entered the ALL-XH-99 clinical trial. Kaplan-Meier method was used to estimate survival rates and differences were compared with the 2-sided log-rank test, statistics was done by SPSS.</p><p><b>RESULTS</b>Out of the 158 patients, 153 (96.8%) attained complete remission (CR) in a median time of 33 days. The overall EFS rate at 5 years was (72.4 +/- 7.8)% with a median observation duration of 26 months. The EFS rates at 5 years in low-risk (LR), median-risk (MR) and high-risk (HR) groups were (88.9 +/- 5.5)%, (78.5 +/- 8.0)% and (53.4 +/- 10.9)%, respectively (P < 0.05). Relapse occurred in 15 patients (10.0%) in a median time of 12 months, including 13 isolated hematologic relapses, 2 isolated central nervous system (CNS) relapses. Seven patients died of complications, and 13 died of leukemia relapse.</p><p><b>CONCLUSION</b>The early response to therapy was an important independent prognostic factor, high-dose methotrexate (HD-MTX) was effective for preventing haematological and testicular relapse. The ALL-XH-99 protocol decreased the rate of therapy-related death and improved the long-term event-free survival rate.</p>

Response rate of AML-XH-99 protocol in the treatment of 82 childhood acute myeloid leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the outcomes of childhood acute myeloid leukemia (AML) treated with AML-XH-99 protocol and explore how to improve the event-free survival (EFS) rate.</p><p><b>METHODS</b>Eighty-two patients entered AML-XH-99 clinical trial. Survival rates were evaluated by Kaplan-Meier method with SPSS.</p><p><b>RESULTS</b>Among the 82 patients, 58 (70.7%) achieved complete remission (CR) after one course treatment, and the total CR rate was 84.1%. The overall 5 year EFS rate was (46.1 +/- 9.1)% and disease-free survival (DFS) rate was (54.3 +/- 10.3)% over a median observation period of 23 months. The 5 year EFS rate of 56 patients received high-dose cytarabine(HD-Ara-C) as intensification therapy was (47.2 +/- 12.9)%. Relapse occurred in 19 patients (26.0%) with a median time of 10 months (ranges 2 approximately 53 months), 28 patients died.</p><p><b>CONCLUSION</b>AML-XH-99 protocol resulted in a higher CR rate, especially one course CR rate, which was one of the key factors for long-term EFS and HD-Ara-C intensification therapy was effective in the treatment of childhood AML.</p>

Treatment of advanced malignant solid tumors in children with autologous hematopoietic stem cell transplantation / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the efficacy of autologous hematopoietic stem cell transplantatation in children with advanced solid tumors.</p><p><b>METHODS</b>Bone marrow was collected from the anterior crista of iliac in both sides in 13 cases while peripheral mononuclear cell was harvested with CS-3000 cell separator in other 15 patients after G-CSF mobilization. Since one of them was suspected to have bone marrow involvement of the neuroblastoma cells, the autograft was purged with CliniMACS based on the CD34 positive selection. Conditioning regimen was CBV protocol (cyclophosphamide + bischloro-nitrosourea, i.e., BCNU + etopside) in two children with Hodgkin's disease and etopside plus carboplatin plus melphalan in others.</p><p><b>RESULTS</b>The number of mononuclear cells collected from bone marrow or peripheral blood was equal to (5.4 +/- 2.1) x 10(8)/kg and (4.1 +/- 1.9) x 10(8)/kg, respectively. Hematopoietic reconstitution was achieved in all patients. Mean time of the neutrophil count recovery to 0.5 x 10(9)/L and mean time of platelet recovery over 2.0 x 10(9)/L were 11.8 +/- 5.7 and 21.0 +/- 9.3 days, respectively. Three units of packed red blood cells and three units of platelet products were transfused in the course of transplantation on average. A total of 12 children developed neutropenic fever and 3 of them had positive blood culture, including staphylococcus epidermal, staphylococcus saprophyte and bacillus subtilis. None of the children died of transplantation-associated complication. One child developed acute renal failure, pulmonary edema and pericardial effusion followed by respiratory distress syndrome. Mechanical ventilation and pulmonary surfactant were used and the patient recovered at last. Another patient developed BCNU associated pulmonary injury, severe pulmonary hypertension and eosinophilosis and recovered after treatment. The mean follow up time was 13 months. Among the 27 children, five died of relapse 5 months after transplantation, and one case of NHL had CNS involvement 3 months after transplantation but has got 17 months of survival till now. The remaining twenty one children were in status of disease-free survival.</p><p><b>CONCLUSION</b>Autologous stem cell transplantation might be effective in the treatment advanced solid tumors in children.</p>

Characterization of CD10 expression and its significance in minimal residual disease detection in childhood B-acute lymphoblastic leukemia / 中国实验血液学杂志

To observe the expressions of CD10 in childhood B-acute lymphoblastic leukemia (B-ALL) and to define the role of CD10 in minimal residual disease (MRD) detection. 58 cases of childhood B-ALL were studied in this program. Four-color flow cytometry was used to analyze the characteristics of B-ALL phenotypes. The four-color fluorochrome labeled antibody combinations of CD10 with other markers were used to detect MRD. The results showed that CD10 overexpression (CD10(bright)) was detected in 65.5% (38/58) of B-ALL patients and a strong correlation between CD10(bright) and CD34 expression was also observed, i.e. CD10(bright) expression most frequently happened in B-ALL with high percentage of CD34 positive cells. In detection of MRD, CD10(bright), combined with other markers, could effectively distinguish normal cells with leukemic cells, even if there was no any other marker that can be used. It is concluded that CD10(bright) expression correlates with high expression of CD34 in B-ALL, it is a good marker for MRD detection. The combination of CD10 and other markers can be applied in B-ALL MRD detection with flow cytometry.

Detection of minimal residual disease in childhood B-ALL by flow cytometry / 中华血液学杂志

<p><b>OBJECTIVE</b>To establish a flow cytometric method for detecting minimal residual disease (MRD) in children with B-ALL and evaluate its clinical application.</p><p><b>METHODS</b>Fifty-eight childhood B-ALL cases entered this study and 30 MRD analyses were performed after remission induction therapy. Four-color fluorochrome labeled monoclonal antibodies were used to analyze the cell immunophenotypes. Cells from normal bone marrow were used as controls. The leukemic cell populations located in flow cytometry dot plots different from those of normal were considered to be the markers of interest in the first step screening, and then used to monitor MRD step after therapy.</p><p><b>RESULTS</b>Fifty-eight cases of childhood B-ALL were screened for antibodies combinations of interest and were identified in 89.7% (52/58) of these cases. The four-color antibody combinations consisted of CD(10)/CD(34)/CD(19) plus another effective marker such as CD(38), CD(65), CD(66c), CD(21). The sensitivity of this method was 0.01%, much higher than microscopic inspection. In 8 cases whose bone marrow microscopically showed no residual leukemic cells, the percentage of leukemic cells were identified with this method of 0.028%, 1.430%, 3.050%, 0.015%, 5.660%, 2.700%, 0.027%, and 0.069%, respectively.</p><p><b>CONCLUSION</b>The application of flow cytometry in MRD monitoring can significantly improve the detection sensitivity in childhood B-ALL, thus facilitate the further treatment decision and follow-up.</p>

Application of CD34+ autologous peripheral progenitor cell transplant in the treatment of children with refractory SLE / 中华儿科杂志

<p><b>OBJECTIVE</b>Systematic lupus erythematosis (SLE) is a severe disease which affects the patient for many years and there is no radical cure for the disease. To explore a possible way to treat children with refractory SLE, the authors treated 2 children with grade III and IV lupus nephropathy for 5 years and 7 years respectively, mainly presented with persistent thrombocytopenia, proteinuria, pleural effusion with CD34(+) autologous peripheral progenitor cells transplantation.</p><p><b>METHODS</b>Mobilized with G-CSF and collected with CS-3000 Cell Separator, passed through the CliniMacs CD34(+) cell selection device, the count of CD34(+) cells obtained reached 1.0 x 10(6)/kg and 1.7 x 10(6)/kg, respectively with the remaining of 2.0 x 10(5)/kg and 1.0 x 10(4)/kg of CD3(+) cells individually. The selected CD34(+) cells were frozen at -80 degrees C. The conditioning regimen consisted of cyclophosphamide [50 mg/(kg x day) for 4 days] plus ATG [Fresennius S 5 mg/(kg x day) for 3 days]. After 48 h treatment with cyclophosphamide, the frozen stem cells were infused back to the patients.</p><p><b>RESULTS</b>Neutrophils recovered on 9 and 7 days after transplantation respectively in these 2 cases. Beginning from 15 days, the platelet count recovered and remained at over 100 x 10(9)/L. The sign of Cushing's syndrome disappeared completely 3 months after transplantation because discontinuing the steroid. One child's height had a 5 cm increase within 6 months after stopping steroid and this was the first height gain during the 7 years since she had had the disease. Till this paper was written, these 2 children were followed up for 13 months and 6 months, respectively, all the original symptoms and autoantibodies related to autoimmune disorders disappeared. But the cell-mediated immunity did not recover yet with the CD4(+) cell level still remained at a lower level.</p><p><b>CONCLUSION</b>The effect of CD34(+) autologous peripheral progenitor cell transplantation on the children with refractory SLE was satisfactory so far, but the long-term effect remains to be confirmed by further studies on more cases.</p>