Three cases of occupational acute trimethyltin chloride poisoning / 中华劳动卫生职业病杂志

Trimethyltin chloride is a highly toxic substance, which is absorbed through respiratory tract, skin and digestive tract, with central nervous system injury as the main clinical manifestations, and can be accompanied by damage to various organs. In this paper, the treatment process of 3 patients with acute trimethyltin chloride poisoning was reviewed, and their clinical manifestations, auxiliary examination, diagnosis and treatment were analyzed. Three patients were misdiagnosed as mental abnormality, encephalitis, and hepatic encephalopathy in different hospitals in the early stage of medical treatment, suggesting that clinicians should pay attention to the occupational contact history of poisoned patients and conduct toxicant detection in time to avoid misdiagnosis and mistreatment.

Impact of Coronavirus Disease 2019 on Prognosis and Treatment Strategies in Patients with Hematological Malignancy--Review / 中国实验血液学杂志

Coronavirus disease 2019 (COVID-19) has spread rapidly worldwide since outbreak in December 2019, and become a global public health crisis. Patients with hematological malignancy concurrently infected with COVID-19 are often associated with severe even fatal complications, due to low basic immune function, high intensity of chemotherapy and radiotherapy, and slow immune reconstruction post hematopoietic stem cell transplantation, and their treatment strategies, such as anti-infective therapy, blood transfusion, and the use of granulocyte colony stimulating factor need to be adjusted. The characteristics of patients, chemotherapy, hematopoietic stem cell transplantation, and other clinical factors may affect the prognosis of patients with hematological malignancy concurrently infected with COVID-19. Herein, the latest research progress is reviewed.

Research progress of circumferential tracheal reconstruction via tissue-engineered trachea / 中华外科杂志

Tissue engineering, as a new technology, provides a new avenue for the reconstruction of circumferential tracheal defects, which has always been a tremendous challenge for surgeons around the world. Recently, technologies such as decellularization, 3-dimensional printing, electrospinning and cell sheet have significantly enhanced the chondrification. Implantation of epithelial cells or transplantation of epithelial cell sheets also has accelerated the process of epithelialization. And pedicle muscle flap proved to be a reliable strategy for vascularization of tissue-engineered trachea. But it is still a huge challenge to achieve circumferential tracheal functional reconstruction. The key difficulty lies in how to simultaneously realize the functional regeneration of cartilage, blood vessels and epithelial tissues of tissue-engineered trachea. Therefore, how to integrate the above schemes and finally realize segmental tracheal reconstruction needs further research. This article reviews the research progress of repairing circumferential tracheal defects based on tissue engineering technology.

Research Advances on Clinical Application of Umbilical Cord Blood Transplantation in the Treatment of Hematological Diseases--Review / 中国实验血液学杂志

Abstract　　Umbilical cord blood (UCB) is an alternative source of hematopoietic stem cells (HSCs) for patients who were lack of HLA match related or unrelated donors. Compared with bone marrow and mobilized peripheral blood, UCB has the advantages of easy availability, safety for donors, and low requirement for HLA match between donors and recipients. However, the cell amount in UCB is relatively less, which was associated with increased graft failure, delayed hematologic recovery, immune reconstitution, and higher transplant related mortality after UCB transplantation (UCBT). Double-unit UCB is a straightforward method to augment cell amount in UCB. Compared with single-unit UCBT, double-unit CBT associated with less risk of primary disease relapse and increased incidence rate of graft-versus-host disease (GVHD), but the hematologic recovery and overall survival of recipients were no significantly difference between single and double-unit UCBT. Novel strategies for UCB expansion significantly increased the cell amount in UCB, single-unit expanded UCBT not only increased the sources of UCB, but also decreased the high cost of double-unit UCB. ATG can decrease the risk of graft failure and GVHD rate, but the role of ATG in UCBT is still controversial. Herein, the recent clinical advances on UCBT in the treatment of hematologic diseases are systematically reviewed.

Bioinformatic Analysis of Related Genes and Therapeutic Drugs of Asthma / 中国实验方剂学杂志

Objective::Bioinformatic analysis was used to compare the gene expression profile between asthma patients and healthy people, and the gene characteristics of asthma were preliminarily identified and the potential mechanism and drugs were revealed. Method::The GSE74986 gene expression profile was downloaded from the gene expression omnibus (GEO) and the differentially expressed genes (DEGs) were analyzed by GEO2R. Then the gene heat map of DEGs was made by Morpheus, and their gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analysis were performed by DAVID 6.8. Moreover, the protein-protein interaction (PPI) network and hub genes were constructed by String 10.5. Finally, the significant modules were analyzed by MCODE in Cytoscape 3.6.1, small molecule drugs related to asthma were screened through Coremine Medical. Result::A total of 510 DEGs were screened, including 29 up-regulated genes and 481 down-regulated genes. DEGs were mainly involved in these biological processes and pathways, including chromatin silencing, transcriptional regulation of RNA polymerase Ⅱ promoter, protein transport, messenger RNA (mRNA) processing, RNA splicing, ubiquitin-mediated proteolysis, protein processing in the endoplasmic reticulum, RNA transport, and myeloid differentiation factor (MyD)-dependent Toll-like receptor signaling pathway, platelet activation, nucleotide binding oligomerization domain (NOD)-like receptor signaling pathway and so on. A total of 9 hub genes were obtained, including T-complex protein 1 subunit theta (CCT8), T-complex protein 1 subunit alpha (TCP1), 26S protease regulatory subunit S10B (PSMC6), heat shock protein 90 alpha (HSP90A)A1, cell cycle protein C (CCNC), HSP90AB1, 26S proteasome non-ATPase regulatory subunit 6 (PSMD6), ubiquitin-specific protease 14 (USP14) and eukaryotic translation initiation factor 4E (EIF4E). Two important modules were obtained. The genes in two modules mainly involved these biological process, such as splice, ubiquitin-mediated proteolysis, protein modification, RNA modification and so on. Some potential molecular drugs for the treatment of asthma, such as anisomycin and genistein, have been developed. Conclusion::DEGs and hub genes can contribute to understanding the molecular mechanism of asthma and providing potential therapeutic targets and drugs for the diagnosis and treatment of asthma.

Analysis of Clinical Pathological Features and Prognosis in Young Patients with Diffuse Large B Cell Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To explore the clinical pathological features of the patients with diffuse large B cell lymphoma (DLBCL) and their prognostic factors.@*METHODS@#The prognosis of the clinical pathological features and their influence on prognosis of 177 patients diagnosed as DLBCL at the first visit from January 2013 to May 2017 in our hospital were analyzed retrospectively.@*RESULTS@#The univariate analysis showed that overall survival (OS) and progression-free survival (PFS) were associated with later Ann Arbor stage (Ⅲ-Ⅳ) ( P＜0.01, P＜0.05), high performance status (ECOG score 2-4) (P＜0.01, P＜0.05), extranodal involvement ＞1 (P＜0.01, P＜0.05), elevated LDH level (P＜0.01, P＜0.05). B symptom (P＜0.05) and elevated β2-MG level (P＜0.05) also influenced OS. COX multivariate analysis showed that the elevated β2-MG level (P＜0.05) and later stage (Ⅲ-Ⅳ) (P＜0.05) have an independent influence on OS, later stage (Ⅲ-Ⅳ) (P＜0.05) also independently influenced PFS. The patients with high aaIPI score (2-3) and bone marrow involvement before treatment had poor OS (P＜0.01, P＜0.01) and PFS (P＜0.05, P＜0.01).@*CONCLUSION@#Elevated β2-MG level can independently influence OS, and later stage (Ⅲ-Ⅳ) can independently influence both OS and PFS. High aaIPI score (2-3) and bone marrow involvement before treatment have an inferior influence on OS and PFS.

Analysis on Mechanism of Shema Zhichuan Liquid in Treating Asthma Based on Network Pharmacology / 中国实验方剂学杂志

Objective: To explore the potential mechanism of Shema Zhichuan liquid in treatment of asthma by network pharmacology. Method: Bioinformatics analysis tool for molecular mechanism of traditional Chinese medicine (TCM), systematic pharmacological database and analysis platform of TCM were employed to find the components in Shema Zhichuan liquid and their targets, and asthma-related genes were obtained from the comparative toxicogenomics database (CTD). The data set of Shema Zhichuan liquid-gene and asthma-gene were imported into the Draw Venn Diagram for intersection analysis. The obtained data set of Shema Zhichuan liquid-asthma-gene was imported into String 11.0 for protein-protein interaction (PPI) analysis, and was visualized by Cytoscape 3.6.1, and further important modules were analyzed with MCODE. DAVID 6.8 was used to analyze pathway enrichment and biological process of Shema Zhichuan liquid-asthma-gene. Result: A total of 399 components and 2 099 potential targets were obtained from Shema Zhichuan liquid, 98 asthma-related targets were retrieved, 45 common genes and 16 hub genes were screened, including transforming growth factor-β1(TGF-β1), heme oxygenase-1 (HMOX1), interleukin-4 (IL-4), etc. Enrichment analysis showed that the common biological processes of Shema Zhichuan liquid and asthma were related to inflammation, contraction and remodeling of airway, cell proliferation and apoptosis, etc. The common biological pathways mainly included tumor necrosis factor (TNF) signaling pathway, receptor with high affinity for immunoglobulin E (Fc epsilon RI) signaling pathway, nuclear transcription factor-kappa B (NF-κB) signaling pathway, nucleotide binding oligomerization domain (NOD)-like receptor signaling pathway and so on. Conclusion: Shema Zhichuan liquid serves as a multi-target, multi-pathway treatment for asthma, which can provide a reference for the further research and clinical application of this preparation.

Analysis of malaria cases in Haidian District of Beijing from 2005 to 2017 / 中华疾病控制杂志

Objective To understand the epidemiological character of malaria in Haidian District of Beijing from 2005 to 2017. Methods The epidemiological data of malaria was collected from the infectious disease reporting system of medical institutions at various levels in Haidian District of Beijing from 2005 to 2017, and the epidemiological methods was used to analyze the distribution of malaria in population, time and region. Results From 2005 to 2017, 111 malaria cases were reported in Haidian District of Beijing, the annual average incidence rate was 0.26/100 000 and one death case was reported in 2014. Among the four reported types of falciparum malaria, vivax malaria, three-day malaria and untyped malaria, the most common falciparum malaria (54.5%, 60/111), no mixed infection; The peak incidence was concentrated in the summer and autumn of June-September (52.0%, 58/111); the cases were mainly occurred in young adults aged from 20 to 59(93.7%,104/111), and the incidence of males was higher than that of females ( 2=52.9, P<0.001); Cadres were the main ward population (33.3%, 33/111). Malaria cases were reported in 26 streets and towns in Haidian District. 81 cases were imported from abroad, accounting for 71.4% of the total cases, of which 74 (91.36%) were originated from Africa. Conclusions In the past 13 years, the incidence of malaria was sporadic, mainly in imported cases. The monitoring of malaria should be strengthened by entry and exit to prevent the second-generation cases of malaria.

Ressarch Advances of Prevantive and Therapeatic Effect of Mesenchymal Stromal Cells on Graft-Versus-Host Disease--Review / 中国实验血液学杂志

Graft-versus-host disease (GVHD) is a frequently encountered serious complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT), it limits the success and widespread use of allo-HSCT. Mesenchymal stem cells (MSCs) are selected as ideal prophylactic and treatment means for GVHD during allo-HSCT due to their unique immunomodulatory and regenerative properties. Herein, the recent research progress about the prevantive and therapeutic effects of MSCs on GVHD and several issues related with the applications of MSC, including whether MSCs increasing risk of primary disease relapse and infection, impact of several clinical parameters on the clinical response to MSCs, and the prevantive and therapeutic effect of MSC-derived extracellular vesicles on GVHD are systematically reviewed.

Establishment of A20 Murine B Lymphoma Transplantation Model Labeled with Luciferase / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To establish the animal model of luciferase-transfected A20 murine B cell lymphoma, so as to provide experimental tools to explore the effect of graft versus tumor.</p><p><b>METHODS</b>Luciferase- labeled A20 cells were cloned with puromycin selection. Transfected A20 cells and CBL/6 bone marrow were inoculated into the irradiated BALB/c mice by injection in tailvein to establish the transplantation model. The bioluminescent imaging technique was used to monitor the tumor growth, and then the survival, body weight, tumor formation and pathological characteristics of target organs were observed.</p><p><b>RESULTS</b>A20 cell line stably expressing luciferase gene was successfully obtained. The the bioluminicent imaging found that the tumor luminescence could be observed on day 8 of A20 cell inoculation, and the mean fluorescent intensity was increased along with the tumor growth. Compared with the BMT group, the survival rate and body weight of BMT+A20-Lucmice were decreased significantly. General anatomy showed the tumor mainly formed in the liver and spleen.</p><p><b>CONCLUSION</b>A mouse transplantation model with luciferase- transfected A20 cells has been successfully established, thus laying a foundation for investigation of graft-versus-tumor.</p>

Effect of G-CSF in vitro Stimulation on Distribution of Peripheral Lymphocyte Subsets in the Healthy Persons / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of granulocyte-colony stimulating factor (G-CSF) in vitro stimulation on the distribution of lymphocyte subset in healthy human.</p><p><b>METHODS</b>Peripheral blood mononuclear cells (PBMNCs) were collected from 8 healthy volunteers by density gradient centrifugation on Ficoll-Paque. In vitro 200 ng/ml G-CSF or 200 ng/ml G-CSF plus 10 µg/ml ConA directly act on PBMNCs, then the colleted cells were cultivated for 3 days. Lymphocyte subsets were stained with the corresponding fluoresce labeled antibodies and detected by flow cytometry.</p><p><b>RESULTS</b>The levels of T cells in G-CSF group and G-CSF+ConA group were both higher than that in the control group (P<0.001, P<0.05). However, there were not significantly different in B cells and NK cells levels among the 3 groups. Furthermore, analysis of the effect of G-CSF on T cell subsets indicated that the levels of CD4T cells and CD8T cells in G-CSF group were both significantly higher than those in control group (P<0.01, P<0.05), Treg cells was not different between G-CSF and control group. Compared with the control group, the level of CD4T cells, CD8T cells and Treg cells in G-CSF+ConA group significantly increased (P<0.05, P<0.01, P<0.01). Analysis of G-CSF receptor (G-CSFR) expression showed that G-CSFR expression on T cells in G-CSF+ConA group dramatically increased, as compared with control group (P<0.01).</p><p><b>CONCLUSION</b>The levels of CD4T cells and CD8T cells in healthy human peripheral blood can be increased by G-CSF stimulation. ConA can enhance the level of T cells and induce G-CSFR expression on T cells.</p>

Role of Intestinal Microbiota in Hematopoietic Stem Cell Transplantation - Review / 中国实验血液学杂志

Graft-versus-host disease (GVHD) and infection are the frequently encountered complications after hematopoietic stem cell transplantation (HSCT), which influence the outcome and limit the widespread application of HSCT. Intestinal microbiota plays an important role in maintaining intestinal immune balance. The diverse levels of intestinal microbiota associated with the incidences of GVHD, infection and the prognosis of HSCT, thus remodeling intestinal microbiota can alleviate GVHD and infection after HSCT. Herein, the recent research progress about the role and the involved mechanisms of intestinal microbiota in HSCT, and the novel manipulation strategies of intestinal microbiota are systematically reviewed.

Arthroscopic reconstruction of anterior cruciate ligament with femoral tunnel made by three-portal technique / 中华骨科杂志

Objective To evaluate the clinical outcomes of arthroscopic reconstruction of anterior cruciate ligament (ACL) with femoral tunnel made by three-portal technique.Methods From February 2014 to May 2015,fifty-five ACL rupture patients (38 males and 17 females) with mean age 31.46±7.34 (range,16 to 48) who underwent single-bundle ACL reconstruction with femoral tunnel made by three-portal technique by the same surgeon were followed up.The average preoperative duration was 15.39±18.57 (range,5 to 180) days.All patients presented the positive Lachman test,of which 32 patients had 1 grade pivot-shift test,15 patients 2 grade and 8 patients 3 grade.At final follow-up,the International Knee Documentation Committee (IKDC) and Lysholm scores were used to estimate knee joint function,while the Lachman test and pivot-shift test were used to estimate knee joint stability.Results All patients were followed up for 26.73±5.23 (range,21 to 34) months.The femoral tunnel positions were accurate without fracture of the back walls in all patients.The mean length of femoral tunnel was 38.58±3.76 (range,35 to 42) mm,and the mean angles of femoral tunnel on coronal plane was 49.32°±2.45° (range,45° to 52°).All patients presented negative Lachman test.Fifty-one patients presented negative pivot-shift test,while the remaining 4 patients had 1 grade of the rotational lax-ity.Forty-nine patients showed normal range of motion,while two had 5° flexion deficit,one had less than 5° flexion deficit and three had 5° hyper-extension loss.Furthermore,the IKDC score significantly was improved from 73.31 ±5.83 preoperatively to 94.63±3.16 at the latest follow-up (t=10.14,P＜0.001).The Lysholm score significantly was increased from 66.35±8.91 preoperatively to 95.32±4.25 at final follow up (t=12.76,P＜0.001).Condusion Arthroscopic reconstruction of ACL with femoral tunnel made by three-portal technique is an reliable technique with the advantage of simple procedure and accurate femoral tunnel position in surgery.

Clinical Characteristics, Treatment and Prognosis of PTLD after allogeneic Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the clinical characteristics of patients with post-transplantation lymphoproliferative disease (PTLD) after allogeneic peripheral blood hematopoietic stem cell transplantation, and to improve the understanding and diagnosis of PTLD.</p><p><b>METHODS</b>The clinical data of 244 patients underwent allogeneic hematopoietic stem cell transplantation in the General Hospital of PLA from May 2014 to April 2017 were analyzed retrospectively. The follow-up time was up to November 30, 2017. The incidence, risk factors, treatment and survival of patients with PTLD were statistically analyzed.</p><p><b>RESULTS</b>Among the 244 cases the PTLD occurred in 22 cases, the incidence rate was 9.02%, 5 of them were diagnosed by pathology, and 17 were diagnosed clinically. All of them had EB virus infection. They were all ATG user, either underwent related haploidentical hematopoietic stem cell transplantation or unrelated hematopoietic stem cell transplantation, 20 cases were treated with rituximab or rituximab combined with γ-globulin, glucocorticoid, ERV+CTL, chemotherapy and 17 showed the effective response, with a total effective rate of 85%. The median follow-up time was 122 days, the median survival time was 5 months (1-22 months) and the total survival rate was 50%.</p><p><b>CONCLUSION</b>The incidence of PTLD after allogeneic peripheral blood hematopoietic stem cell transplantation closely relates with EB virus infection. The application of ATG in the preconditioning scheme is a high risk factor for the onset of PTLD. In the case of no pathological diagnosis, clinical and laboratory examinations should be actively combined so as to define clinical diagnosis. The riturimab should be used more and more for patients with PTLD.</p>

Research Advances on BK Virus Associated Hemorrhagic Cystitis after HSCT-Review / 中国实验血液学杂志

BK virus（BKV）associated hemorrhagic cystitis is one of the most common complications after hematopoietic stem cell transplantation（HSCT）, which can increase the suffering, become time consuming and even life threatening. However, BKV infection has not attracted enough attention in clinical practice. Accurate BKV monitoring and effective clinical treatments are still in urgent need. In recent years, BKV-DNA monitoring has shown a significant clinical value in BKV-associated hemorrhagic cystitis after HSCT. As for clinical treatments, traditional antivirus agent cidofovir is still to show promising effects, in the meantime, the virus-specific lymphocytes and other new therapies are continuously appearing. Here, the most recent advances on pathogenesis, treatment, virus monstoring and so on for BKV associated hemorrhagic cystitis after HSCT are reviwed.

An analysis of donor semen quality in Chongqing Human Sperm Bank / 中华男科学杂志

<p><b>Objective</b>To evaluate the quality of the donor semen in Chongqing Human Sperm Bank and the influence of age on semen parameters.</p><p><b>METHODS</b>We collected semen samples from 899 donors in Chongqing Human Sperm Bank and divided them into five groups according to the age of the semen donors: 22－25, 26－30, 31－35, 36－40, and >40 years old. Using the Makler Counting Chamber, we measured the semen volume, percentage of progressively motile sperm (PMS), total motile sperm, sperm concentration, total sperm count per ejaculate, and percentage of morphologically normal sperm (MNS). Then, we compared the semen parameters obtained with the fifth percentile and median reference values published in the WHO Laboratory Manual for the Examination and Processing of Human Semen-5th Ed (WHO 5th Ed) and among different age groups using the Kruskall-Wallis H test.</p><p><b>RESULTS</b>The semen volume (1.8 ml), sperm concentration (25.0 × 10⁶/ml), total sperm count (100.7 × 106/ejaculate) and MNS (4.3%) in the semen samples of the 899 donors were obviously higher than the fifth percentile values published in the WHO 5th Ed, and so were the first three parameters (4.0 ml, 88.0 × 10⁶/ml, and 333.7 × 106/ejaculate) than the WHO median reference values. PMS (31.0%) and total motile (38.0%) were lower than the WHO fifth percentile values and so was MNS (11.6%) than the WHO median reference value. PMS (55.0%) and total motile sperm (61.0%), however, were coincident with the median reference values of WHO 5th Ed. Statistically significant differences were observed among the 22－25, 26－30, 31－35, 36－40 and >40 years old groups in perm concentration (88.0 ［1.0－270.0］ vs 96.0 ［5.0－335.0］ vs 100.0 ［3.0－200.0］ vs 105 ［15.0－225.0］ vs 90.0 ［22.0－159.0］ × 10⁶/ml, P < 0.05), but not in the semen volume, PMS, total sperm motility, total sperm count or MNS (P > 0.05).</p><p><b>CONCLUSIONS</b>The donor semen in Chongqing Human Sperm Bank is generally of high quality. Sperm concentration significantly increases with age but decreases in men aged >40 years.</p>

Bacterial culture of donor semen: Analysis of results / 中华男科学杂志

<p><b>Objective</b>To investigate bacterial infection and the distribution of different bacterial species in the donor semen and the influence of different bacterial counts on semen quality.</p><p><b>METHODS</b>Bacterial colonies in the semen samples from 1 126 donors were counted with the Synbiosis Protocol 3 Automatic Colony Counter and the bacterial species with a colony count ≥10⁴ cfu/ml identified with the VITEK2 Compact Automatic Biochemical Analyzer. The Makler Sperm Counting Board was used to examine the semen quality of the semen samples with a colony count = 0 cfu/ml (n = 22, group A), those with a colony count <10⁴ cfu/ml (n = 22, group B) and those with a colony count ≥10⁴ cfu/ml (n = 22, group C). Univariate analysis was employed for comparison of semen quality among different groups.</p><p><b>RESULTS</b>Among the 1 126 donor semen samples cultured, 5 (0.44%) showed mixed bacterial contamination and 993 (88.58%) showed none but with growth of a certain species of bacteria, 2.22% (22/993) with a colony count ≥10⁴ cfu/ml, mainly including Streptococcus bovis, tiny bacilli, Staphylococcus epidermis, and Staphylococcus aureus, among which gram-positive and gram-negative bacteria accounted for 95.45% (21/22) and 4.54% (1/22), respectively. Compared with group A, groups B and C manifested significantly reduced total sperm count (［567.5 ± 327.6］ vs ［421.9 ± 155.9］ and ［389.9 ± 110.6］ × 106 per ejaculate, P <0.05) and percentage of progressively motile sperm (［65.0 ± 6.5］ vs ［61.0 ± 3.5］ and ［61.6 ± 4.3］ %, P <0.05). There were no statistically significant differences among the three groups in the semen liquefaction time, semen pH value, total sperm motility or percentage of morphologically normal sperm (P > 0.05). Of the 284 randomly selected semen samples, 34 (11.97%) were found positive for Ureaplasma urealyticum (UU) and no significant difference was observed in the semen quality between the UU-positive and UU-negative samples (P> 0.05).</p><p><b>CONCLUSIONS</b>The bacteria-positive rate is high in the donor semen and the bacterial species are varied, mainly including gram-positive bacteria. Semen quality is reduced with the increased number of bacterial colonies.</p>

Application of Chimeric Antigen Receptor-Modified NK Cells in Multiple Myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the killing effect of CAR (CD138-CD28-CD3ζ)-NK cells on myeloma cells through construction of CAR(CD138-CD28-CD3)-NK cells.</p><p><b>METHODS</b>The antiCD138scFv-CD28-CD3 zeta plasmid pcDNA3.1 was constructed, which then together with 3 plasmid lentiviral packaging system were transfected into 293T cells, the virus was collected. Furthermore, in order to get the stably transfected cell line, the NK92MI cell line was infected by the virus, then the positive cells were screened by puromycin. The expression of the CARNK cells were verified by RT-PCR and Western blot. At last the ability of secreting cytokine CD107a was detected by flow cytometry, and the statistical analysis was carried out to verify the anti-myeloma effect of CAR-NK cells.</p><p><b>RESULTS</b>Gene fragment of the CAR(antiCD138scFv-CD28-CD3ζ) was constructed successfully by gene engineering technique in vitro, and the gene sequence was verified to be correct by sequencing. By virus packaging technology, the virus expressing the protein of the CAR was obtained. PCR and Western blot verified the expression of CAR fusion protein on the sufurce of NK cells. The cell killing experiment confirmed that the CAR-NK cells possessed the ability to secrete cytokine CD107a superior to control cells and showed the obvious killing effect on multiple myeloma cells.</p><p><b>CONCLUSION</b>The CAR can be constructed in vitro, and express on NK92 cells. The CAR-NK cells can kill the multiple myeloma cells expressing CD138 antigen, thereby plays an antimyeloma effect.</p>

Analysis of Hematopoietic Reconstitution in Auto-Peripheral Blood Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the factors which may have influences on hematopoietic reconstitution of the auto-peripheral blood stem cell transplantation(auto-PBHSCT).</p><p><b>METHODS</b>The successful rate, the time of hematopoietic reconstitution and implantation status at 28 days after transplantation of 177 patients received auto-PBSCT were retropectively analyzed, in order to explore the factors which may have influences on hematopoietic reconstitution.</p><p><b>RESULTS</b>The median time of neutrophil recovery was 12 days (8-21 days), implantation rate was 98.9%, all patients' neutrophil were recovered in 28 days. The median time of platelet recovery was 17 days (7-420 days), implantation rate was 95.5%, the cumulative incidence of platelet recovery at day 28 was 80.8%. Univariate analysis showed that the CD34cell number and the use of TPO had effect on neutrophils recovery time; the disease kinds, conditioning regimen and the infused CD34cell number had influence on platelets recovery time. Multivariate analysis showed that the CD34cell number was the independent influencing factor of neutrophils reconstitution time; the disease kinds, the CD34cell number were the independent influencing factors of platelet reconstitution time. Disease kinds and the CD34cell number were the independent influencing factors of hematopoietic reconstitution status of 28 days after transplantation.</p><p><b>CONCLUSION</b>In auto-PBHSCT patients, disease kinds, conditioning regimen, the infused CD34cell number and the use of TPO have been confirmed to be independent influencing factors on hematopoietic reconstitution.</p>

Detection of ATP Level in CD4T Lymphocytes and Its Clinical Significance in Allogeneic Hematopoietic Stem Cell Transplantation Recipients / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical value of detecting adenosine triphosphate (ATP) level in CD4T lymphocytes (Immuknow ATP) of patients on early stage after allogeneic hematopoietic stem cell transplantation (allo-HSCT).</p><p><b>METHODS</b>The base-line ATP value in CD4T lymphocytes in cases of hematological malignancies and the ATP level in CD4T lymphocytes of acute leukemia patients before allo-HSCT were detected. Allo-HSCT recipients were devided into 3 groups with different level of immunereactivity according to ATP concentraiton in month 3 (day 90±5) after allo-HSCT. The clinical characteristics of patients in 3 groups were analyzed.</p><p><b>RESULTS</b>The mass concentration of Immuknow ATP in 15 cases of hematological malignancies before allo-HSCT ranged from 56.21-435.71 ng/ml, with a mean of 203.98±112.72 ng/ml. The ATP level in 46 cases after allo-HSCT ranged from 1.69-333.09 ng/ml, with a median of 41.96 ng/ml. Both 91.26 ng/ml (mean-SD) and 316.70 ng/ml (mean+SD) were used as cutoff, and 36 allo-HSCT recipients (78.3%) were assigned to low immunereactivity group, 8 recipients (17.4%) to middle group and 2 recipients (4.3%) to high group. The incidence of infection in low immunereactivity group was significantly higher than that in middle immunereactivity group (86.1% vs 50.0%)(P=0.022), and also significantly higher than that in high immunereactivity group (86.1% vs 0%)(P=0.002). There were no statistical differences in the incidences of severe infection among 3 groups. The incidence of grade II or higher acute graft versus host disease (aGVHD) in high immunereactivity group was superior to that in low immunereactivity group statistically (100% vs 13.9%)(P=0.002). Immune-mediated organ injury occurred more frequently in high immunereactivity group as compared with low and middle immunereactivity groups (100% vs 0% and vs 0%)(P=0.000; P=0.002). There were no significant differences in relapse rates of leukemia among 3 groups. The percentage of patients with increased trough blood concentration of cyclosporine A(CsA) was not significantly different among 3 groups (P=0.720).</p><p><b>CONCLUSION</b>Detection of ATP level in CD4T lymphocytes on early stage after allo-HSCT possesses clinical significance for predicting infection, severity at aGVHD and immune-mediated organ injury.</p>