OBJECTIVE: To explore the effect and possible mechanism of dimethyl fumarate (DMF) on T-cell acute lymphoblastic leukemia (T-ALL), and provide experimental and theoretical basis for the clinical treatment of T-ALL. METHODS: Jurkat cells were treated with different concentrations of DMF for 24 hours, and then the proportion and absolute count of Ki67-positive Jurkat cells were analyzed by flow cytometry. Meanwhile, the protein levels of nuclear factor-erythroid 2-related factor 2 (Nrf2) and E3 ubiquitin ligase HACE1 in Jurkat cells treated with DMF for 24 hours were evaluated by Western blot. Nrf2 proteins were co-immunoprecipitated in Jurkat cells, and then HACE1 protein was assessed by Western blot. Plasmids of Flag-Nrf2 and different gradients of Flag-HACE1 were transfected into HEK293T cells, and the levels of Flag-Nrf2 were detected by Western blot after 48 hours. RESULTS: DMF could significantly inhibit the proportion and absolute count of Ki67-positive Jurkat cells, and DMF inhibited the proliferation of Jurkat cells in a dose-dependent manner (r=0.9595, r=0.9054). DMF could significantly up-regulate the protein levels of Nrf2 and E3 ubiquitin ligase HACE1 in Jurkat cells (P<0.01, P<0.01). HACE1 physically interacted with Nrf2 in Jurkat cells. Overexpression of Flag-HACE1 significantly increased the protein level of Flag-Nrf2 in a dose-dependent manner (r=0.9771). CONCLUSION DMF inhibits the proliferation of T-cell acute lymphoblastic leukemia cell. The mechanism may be that, DMF significantly up-regulates the protein levels of Nrf2 and E3 ubiquitin ligase HACE1, and HACE1 interacts with Nrf2 and positively regulates Nrf2 protein level.
Dimethyl Fumarate/pharmacology* ; HEK293 Cells ; Humans ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; T-Lymphocytes ; Ubiquitin-Protein Ligases

OBJECTIVE: To observe the genotypes and composition ratio of thalassemia in couples of reproductive age, and provide a reference for the prevention and control of thalassemia in Haikou. METHODS: Gene diagnosis was performed in 2 494 subjects who were screened for thalassemia before marriage or prenatal by cross-breakpoint PCR, PCR-reverse dot hybridization, and PCR-electrophoresis. RESULTS: A total of 1 037 thalassemia gene carriers were detected in 2 494 samples, with a detection rate of 41.57%, of which 75.02% was α-thalassemia, 18.61% was β-thalassemia, and 6.36% was α-β complex thalassemia. There were 778 cases of α-thalassemia, mainly of deletion type, accounting for 76.99% (599/778). Twenty genotypes were detected, the highest three was -- CONCLUSION In Haikou city, the gene carrying rate of thalassemia is very high, and the genotype distribution is different from other cities in Hainan Province, attention should be paid to the impact of population inflow on the frequency spectrum change of local thalassemia gene.
Cities ; Female ; Genetic Testing ; Genotype ; Humans ; Pregnancy ; alpha-Thalassemia/genetics* ; beta-Thalassemia/genetics*

OBJECTIVE: To explore the non-genetic factors of overall survival in patients with multiple myeloma (MM). METHODS: Kaplan-Meier survival curve, Log-rank test and Cox regression model were used to carry out univariate and multivariate retrospective analysis on clinical and laboratory parameters of 51 patients who were newly diagnosed with MM and had complete follow-up data in the Department of Hematology, the Affiliated Jiangning Hospital of Nanjing Medical University from November 2011 to October 2019. RESULTS: Fifty-one patients included 29 males and 22 females. Followed up to December 2019, 21 cases died and 30 cases survived. The univariate analysis showed that the overall survival time of the patients was influenced by age, disease stage, standard treatment, new drugs, maintenance treatment, hypercalcemia, globulin, albumin, and hemoglobin. The overall survival time of patients with age <65 years old, ISS stage I and II, standardized treatment, new drugs, normal or below normal blood calcium, normal or below normal globulin, albumin ≥35 g/L or hemoglobin ≥100 g/L was prolonged significantly (P<0.05). The multivariate analysis showed that maintenance treatment, hypercalcemia (≥2.6 mmol/L), and hemoglobin (<100 g/L) were independent risk factors influencing the prognosis of MM patients. CONCLUSION Patients with blood calcium ≥2.6 mmol/L, hemoglobin <100 g/L, and who do not undergo regular maintenance therapy show a poor prognosis.
Aged ; Female ; Humans ; Male ; Multiple Myeloma/genetics* ; Patients ; Prognosis ; Proportional Hazards Models ; Retrospective Studies

The Wilderness Medical Society updated and published the Wilderness Medical Society practice guidelines for the prevention and treatment of frostbite in July 2019. The guidelines provide evidence-based recommendations for the prevention and treatment of frostbite, mainly including pathophysiology, classification, prevention, and treatment of frostbite. This paper focuses on the interpretation of the guidelines, aiming at making clinical medical staff understand the new progress of frostbite treatment and providing reference for clinical practice.

OBJECTIVETo construct a co-culture system for bone marrow mesenchymal stem cells (BMMSC) and multiple myeloma (MM) cells, and to investigate the effects of co-cultured BMMSC on the migrating and homing of multiple myeloma cells.

METHODSThe BMMSC from the transgenic mice with green fluorescent protein (GFP) fetal bone were cultured by adherent screening. A co-culture system of BMMSC and MM cell line XG-7 cells was constracted, the proliferation and apoptosis of cells were determined by trypan blue exclusion and Annexin V/PI, respectively, MDC staining was employed to detect the autophagy. The moving direction distribution of molecule in BMMSC and XG-7 cells labeled with PE-CD138 in co-culture process were observed dinamically by confocal microscopy.

RESULTSAfter co-culture with GFP-BMMSC, the resistance of XG-7 cells to apoptosis and autophagy were enhanced; at the same time, their proliferation increased. Apoptosis rates of XG-7 cells directly and indirectly co-cultured with BMMSC were (6.23 ± 0.12)% and (6.97 ± 0.03)% respectively, which were lower than that of XG-7 cells cultured alone (17.90 ± 1.46)% (P < 0. 01). There was low level of autophagy in XG-7 cells co-cultured with BMMSC. XG-7 cells are highly polarized and contained a specialized membrane domain with specific protein and lipid components to contact with BMMSC under confocal microscope. After methyl-β-cyclodextrin treatment, the molecules were normally enriched in the specialized domain.

CONCLUSIONBMMSC can protect XG-7 cells from apoptosis and autophagy, and obviously promote the proliferation of XG-7 cells, and can influence the migrating and homing of multiple myeloma cells.

Animals ; Apoptosis ; Autophagy ; Bone Marrow Cells ; cytology ; Cell Line, Tumor ; Cell Movement ; Coculture Techniques ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Transgenic ; Multiple Myeloma ; pathology

This study was purposed to compare the anti-leukemic effects of E.coli-L-Asp and Erwinia-L-Asp in vitro, and to investigate their mechanism. The cell apoptosis and proliferation inhibition rate were measured by CCK-8 kit, and IC50 of two drugs was calculated by using SPSS software. Pro-apoptosis effect of E.coli-L-Asp and Erwinia-L-Asp on REH and Jurkat cell lines was also determined by flow cytometry with Annexin V/PI double staining. Concentration changes of 4 amino acids (Asn, Aspa, Gln, and Glu) before and after medication were detected by using high pressure liquid chromatography (HPLC) assay. The results showed that both REH and Jurkat cell lines were sensitive to L-Asp drugs from two different strains, and E.coli-L-Asp and Erwinia-L-Asp displayed the inhibition effect on the proliferation of Jurkat and REH cell lines in dose-dependent and time-dependent manners. The inhibition cell of proliferation and cell apoptosis in Erwinia-L-Asp group were higher than those in E.coli-L-Asp group after 24 hours (P < 0.05) . However, after treatment of REN and Jurkat cells with 2 kind of L-Asp for 48 hours, the inhibition of cell proliferation and apoptosis rates were not significantly different between the 2 L-Asp drugs (P > 0.05). The Asn in medium could be depleted by two different L-Asp drugs with low concentration. Both the two L-Asp drugs had the same capability to deplete the Asn surrounding leukemia cells (P > 0.05). The Gln in medium could be depleted by two L-Asp drugs with high concentration. The hydrolysis effect of Erwinia-L-Asp on Gln was stronger than that of E.coli-L-Asp (P < 0.05). It is concluded that in a certain range of concentrations, E.coli-L-Asp and Erwinia-L-Asp exert anti-leukemia effect in dose-dependent manner. Depletion of Gln and Asn in surrounding environment and induction of cell apoptosis are two potential mechanisms, by which leukemia cells can be killed. Erwinia-L-Asp may be chosen as the first-line drug to treat childhood ALL for its fast action and stronger hydrolysis effect on Gln.
Apoptosis ; drug effects ; Asparaginase ; pharmacology ; Cell Proliferation ; drug effects ; Flow Cytometry ; Humans ; Jurkat Cells

This study was purposed to investigate the role of post-thaw infused donor cells for predicting engraftment and hematopoietic reconstitution after unrelated cord blood transplantation (UCBT). The retrospective analysis was performed on clinical data of 97 children with malignant or non-malignant diseases received single unit UCBT from August 1999 to April 2010. The impact of pre-freezing and post-thaw cell dose of total nucleated cells (TNC), CD34(+) cells and colony-forming units-granulocyte/macrophage (CFU-GM) on engraftment and hematological recovery after UCBT was analyzed. Unrelated donors were from Guangzhou cord blood bank (GZCBB) entirely. The results indicated that the pre-freezing TNC (/kg) (mean ± SD: 7.65 × 10⁷ ± 4.26 × 10⁷; median: 6.34 × 10⁷), CD34(+)cells (/kg) (mean ± SD: 4.64 × 10(5) ± 4.47 × 10⁵; median: 3.03 × 10⁵) and CFU-GM (/kg) (mean ± SD: 0.79 × 10⁵ ± 1.09 × 10⁵; median: 0.57 × 10⁵) showed a good correlation with their post-thaw counterparts including TNC(/kg) (mean ± SD: 6.98 × 10⁷ ± 4.12 × 10⁷; median: 6.00 × 10⁷), CD34(+)cells (/kg)(Mean ± SD: 6.86 × 10⁵ ± 8.56 × 10⁵; Median: 4.17 × 10⁵), and CFU-GM (/kg) (mean ± SD: 0.52 × 10⁵ ± 0.52 × 10⁵; median: 0.39 × 10⁵) (r = 0.952, p < 0.001; r = 0.794, p < 0.001; r = 0.478, p < 0.001). Either the pre-freezing or post-thaw number of infused CFU-GM was significant higher in patients who achieved engraftment (n = 70) than those who suffered graft failure (n = 22) (p = 0.023 and 0.011, respectively), but no significant difference of TNC and CD34(+) cells dose (pre-freezing or post-thaw) were found between these two groups. Pre-freezing CFU-GM, TNC, CD34(+) cell dose negatively correlated with the time of neutrophil engraftment (r = -0.285, p = 0.018; r = -0.396, p = 0.002; r = -0.373, p = 0.002), as well as the post-thaw number of TNC and CD34(+) cells (r = -0.260, p = 0.031; r = -0.483, p < 0.001), whereas only pre-freezing CD34(+) cells showed a significant correlation with platelet engraftment time (r = -0.352, p = 0.013). It is concluded that the CFU-GM amount is useful for predicting engraftment of UCBT, while pre-freezing hematopoietic cell doses show superior correlation with the speed of engraftment and hematopoietic reconstitution than their post-thaw counterparts in pediatric recipients, suggesting that it is essential to perform hematopoietic potency assay on each cord blood unit prior to listing or release for administration.
Adolescent ; Antigens, CD34 ; blood ; Blood Banks ; Child ; Child, Preschool ; Cord Blood Stem Cell Transplantation ; methods ; Female ; Fetal Blood ; cytology ; Graft Survival ; Granulocyte-Macrophage Progenitor Cells ; Humans ; Infant ; Male ; Retrospective Studies ; Tissue Donors

Umbilical cord blood (UCB) is an alternative source of hematopoietic stem cells for transplantation with success being associated with the total nucleated cell (TNC) count, CD34(+) cells and colony-forming unit-granulocyte-macrophage (CFU-GM) content infused. This study was purposed to clarify the impact of maternal and neonatal factors on hematopoietic potential of UCB product. UCB samples were screened, processed, tested and cryopreserved according to the Standard Operation Procedure (SOP) of Guangzhou cord blood bank (GZCBB). Relationship of hematopoietic cell parameters with maternal and neonatal characteristics for 4615 UCB units was analyzed retrospectively. The results showed that both collected volume (Mean ± SD: 95.23 ± 22.42 ml; Median: 91.85 ml) and initial TNC [Mean ± SD: (1.34 ± 0.49) × 10(9); Median: 1.25 × 10(9)] correlated well with postprocessed TNC [Mean ± SD: (1.21 ± 0.42) × 10(9); Median: 1.14 × 10(9); p < 0.001], CD34(+)count [Mean ± SD: (5.14 ± 4.55) × 10(6); Median: 4.08 × 10(6); p < 0.001] and CFU-GM content [Mean ± SD: (9.72 ± 8.66) × 10(5); Median: 7.53 × 10(5); p < 0.001]. As for donor factors, only infant birth weight correlated strongly with volume collected and all hematopoietic cell parameters (p < 0.001). UCB samples from bigger babies had higher collected volume, TNC, CD34(+) count and CFU-GM content (p < 0.001). Mother's age had no correlation with all the above parameters. Gestational age correlated positively with initial/postprocessed TNC (p < 0.001) and negatively with CD34(+) count (p = 0.04), but no relation with collected volume and CFU-GM content. Cesarean section produced superior volume (Mean ± SD: 97.05 ± 22.23 ml vs 92.53 ± 22.43 ml; Median: 94.08 ml vs 88.82 ml; p < 0.001), but inferior cell count than vaginal delivery (p < 0.001). Male infants had more initial volume and CD34(+) count (Mean ± SD: 96.41 ± 22.31 ml vs 93.95 ± 22.47 ml; Median: 93.27 ml vs 90.14 ml; p < 0.001); [Mean ± SD: (5.28 ± 5.04) × 10(6) vs (5.00 ± 3.94) × 10(6); Median: 4.18 × 10(6) vs 3.94 × 10(6); p < = 0.042], but lower initial and postprocessed TNC than female ones [Mean ± SD: (1.31 ± 0.50) × 10(9) vs (1.37 ± 0.47) × 10(9); Median: 1.22 × 10(9) vs 1.28 × 10(9); p < 0.001]; [Mean ± SD: (1.18 ± 0.42) × 10(9) vs (1.24 ± 0.41) × 10(9); Median: 1.10 × 10(9) vs 1.17 × 10(9); p < 0.001], while no significant difference of CFU-GM were found between male and female infants. It is concluded that these data may be helpful to optimize the UCB donor selection and improve cost efficiency of UCB bank resource. The heavier infants after vaginal delivery should be selected and large-volume units with higher TNC should be chosen at first.
Adult ; Birth Weight ; Blood Banks ; methods ; Cord Blood Stem Cell Transplantation ; methods ; Delivery, Obstetric ; Donor Selection ; Female ; Fetal Blood ; cytology ; immunology ; Gestational Age ; Hematopoietic Stem Cells ; Humans ; Infant, Newborn ; Male ; Maternal Age ; Pregnancy ; Young Adult

P13-OMP (29.1). P13-OMP and OMP68 group challenged with P13 and P11 can be efectivly protected; P13-WCB group challenged with P13 and P11 can not be efectivly protected; the control group were died out. The P13-OMP and OMP68 of Bordetella bronchiseptica has good immunogenicity and protection, so the results of this study lay good theoretical foundation for OMP subunit vaccine.

OBJECTIVEStomach physiological effects of channel tropism of stomach, cold & cool Chinese herbal medicine on rats with stomach-heat syndrome were studied.

METHODUsing water decoction of warm &heat medicine, Rhizoma Zingiberis to feed rats for 15 days continuously, causing the stomach-heat sydrome model, then decoction of Rhizoma Coptidis, Herba Taraxaci, Fractus Aurantii Immaturus was used to feed rats for 10 days, respectively. Biochemical indexes of blood reflecting the physiological function of stomach, including thromboxaneB2 (TXB2), 6-keto-PGF(1alpha), Gastrin (Gas), Motilin (MTL), and Somatostation (SS) were measured by radioimmunoassay.

RESULTSymptom of stomach-heat syndrome prevailed in body of rats after filled with decoction of Rhizoma Zingiberis, values of TXB2/6-keto-PGF(1alpha), MTL, and Gas in blood raised up evidently, compared with the control (P < 0.05), but values of 6-keto-PGF(1alpha). decreased conspicuously (P < 0.05). After treated with decoction of Rhizoma Coptidis, Herba Taraxaci or Fractus Aurantii Immaturus for 10 days, respectively. Symptoms of stomach heat syndrome were eliminated or alleviated, values of 6-keto-PGF(1alpha), and SS in blood elevated at different degrees, and those of TXB2, TXB2/6-keto-PGF(1alpha), MTL, and Gas felled down at different degrees. Difference of efficacy existed at different groups, group of Rhizoma Coptidis was the strongest, group of Herba Taraxaci was the second, group of Fractus Aurantii Immaturus was the third. Efficacy of medicine in groups with high dosage was stronger than those with low dosage.

CONCLUSIONChannel tropism of stomach, cold and cool Chinese herbal medicine could improve the physiological functions of stomach effectively, and the efficacy concerns with the degree of their cold and cool characteristics.

6-Ketoprostaglandin F1 alpha ; blood ; Animals ; Citrus ; chemistry ; Coptis ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; therapeutic use ; Gastrins ; blood ; Ginger ; Male ; Medicine, Chinese Traditional ; Motilin ; blood ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Somatostatin ; blood ; Stomach Diseases ; blood ; chemically induced ; drug therapy ; Syndrome ; Taraxacum ; chemistry ; Thromboxane B2 ; blood