Mechanism of famous classical formula Huaihua Powder in treatment of ulcerative colitis based on metabonomics / 中国中药杂志

Ultra-high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry(UHPLC-Q-TOF-MS) was employed in this study to observe the effect of Huaihua Powder on the serum metabolites of mice with ulcerative colitis and reveal the mechanism of Huaihua Powder in the treatment of ulcerative colitis. The mouse model of ulcerative colitis was established by dextran sodium sulfate salt(DSS). The therapeutic effect of Huaihua Powder on ulcerative colitis was preliminarily evaluated based on the disease activity index(DAI), colon appearance, colon tissue morphology, and the content of inflammatory cytokines such as tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-1β(IL-1β). UHPLC-Q-TOF-MS was employed to profile the endogenous metabolites of serum samples in blank control group, model group, and low-, medium-, and high-dose Huaihua Powder groups. Multivariate analyses such as principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA), and orthogonal partial least squares discriminant analysis(OPLS-DA) were performed for pattern recognition. Potential biomarkers were screened by Mass Profiler Professional(MPP) B.14.00 with the thresholds of fold change≥2 and P<0.05. The metabolic pathways were enriched by MetaboAnalyst 5.0. The results showed that Huaihua Powder significantly improved the general state and colon tissue morphology of mice with ulcerative colitis, reduced DAI, and lowered the levels of TNF-α, IL-6, and IL-1β in serum. A total of 38 potential biomarkers were predicted to be related to the regulatory effect of Huaihua Powder, which were mainly involved in glycerophospholipid metabolism, glycine, serine, and threonine metabolism, mutual transformation of glucuronic acid, and glutathione metabolism. This study employed metabolomics to analyze the mechanism of Huaihua Powder in the treatment of ulcerative colitis, laying a foundation for the further research.

Effect of Recombinant Human Thrombopoietin (rhTPO) on Hematopoietic Reconstitution in Allogeneic Hematopoietic Stem Cell Transplantation Model / 中国实验血液学杂志

OBJECTIVE@#To explore the effect of recombinant human thrombopoietin (rhTPO) on hematopoietic reconstruction in allogeneic hematopoietic stem cell transplantation (allo-HSCT) model.@*METHODS@#The C57BL/6 mice were employed as the donors, and BALB/c mice as recipients. The bone marrow mononuclear cells of the donor mice were extracted and pretreated, which then were injected with 5×106 per mouse through the tail vein of the recipient to establish an allo-HSCT model. The implantation of hematopoietic stem cells in the recipient mice was detected by flow cytometry on the 28th day after transplantation. Next, the successfully modeled recipient mice were randomly divided into experimental group and control group. The rhTPO was injected into mice in the experimental group on the first day after transplantation, while the saline was injected into mice in the control group. Both groups were injected for 14 consecutive days. The peripheral blood and bone marrow hematopoiesis of the two groups were observed on day 1, 3, 7, 14, and 21 after transplantation.@*RESULTS@#The expression rate of H-2Kb in the bone marrow of recipient mice was 43.85% (>20%) on the 28th day after transplantation, which indicated that the recipient mice were successfully chimerized. Meanwhile, counts of PLTs on the day 3, 7, 14, and 21 after transplantation in the experimental group were higher than those in the control group with statistical significances (P<0.05). In addition, hematopoietic function of bone marrow was suppressed in both groups on day 1, 3 and 7 after transplantation, but hematopoietic bone marrow hyperplasia was better in the experimental group than in the control group. On day 14 and 21 after transplantation, the hematopoietic function of bone marrow in the two groups was recovered, and the experimental group showed more obvious than the control group.@*CONCLUSION@#rhTPO can effectively stimulate the production of PLTs and facilitate the recovery of white blood cells and hemoglobin after allo-HSCT, and promote hematopoietic recovery and reconstitution of bone marrow.

Transduction of mesenchymal stem cells with multidrug resistance gene provides protection for bone marrow toxicity after being transplanted into a nude mice model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Myelosuppression is the main dose-related toxicity of many chemotherapeutic drugs. The human multidrug resistance (mdr1) gene is well-known for its ability to confering drug resistance. In this study, we meant to transplant the placenta mesenchymal stem cells (P-MSCs) moderated by mdr1 gene into a nude mice model radiated by γ-Co(60) and to explore the chemoprotection for bone marrow (BM) toxicity.</p><p><b>METHODS</b>Human P-MSCs were isolated from trypsin-digested term placentas and then transduced by with reconstructed retroviral vector containing mdr1 gene and green fluorescent protein (GFP) reporter gene. The integration and expression of mdr1 gene was observed indirectedly by the expression of GFP. A nude mice model was constructed after irradiation with a sublethal dosage of γ-Co(60). These irradiated mice were transplanted with mdr1-MSCs through the caudal vein and then received paclitaxel (PAC) intraperitoneal chemotherapy. The Peripheral peripheral blood (PB) of the nude mice was collected, and the PB cells counts and values were determined using an automatic analyzer.</p><p><b>RESULTS</b>After PAC treatment, mdr1-MSCs transplanted mice showed markedly improved survival upon compared to MSCs transplanted mice (85.7% vs. 57.1%). White blood cell (WBC) and red blood cell (RBC) counts as well as the hemoglobin (Hb) values were significantly increased in PAC treated mdr1-MSCs mice compared to PAC treated control mice when PAC chemotherapy had been finished (all P < 0.05), but the difference was not found in the plateltes (PLT) count (P > 0.05).</p><p><b>CONCLUSION</b>Human P-MSCs moderated by mdr1 gene when transplanted into nude mice may provide chemoprotection for hematopoietic toxicity.</p>

Retroviral-mediated transfer and functional expression of multidrug resistance gene in human placenta mesenchymal stem cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Most of gynecologic malignancies are sensitive to chemotherapy. Myelosuppression is the main dose-related toxicity of many chemotherapeutic drugs. The human multidrug resistance (mdr1) gene is well known for its ability to confer drug resistance. This study aimed to explore the feasibility of expression and resistance of mdr1 gene transduction into human placenta mesenchymal stem cells (P-MSCs) by retrovirus vector.</p><p><b>METHODS</b>Human P-MSCs were isolated from trypsin-digested term placentas, and their immunophenotypes and differentiation potential were evaluated. Human P-MSCs were transduced by reconstructed retroviral vector containing the mdr1 gene and green fluorescent protein (GFP) reporter gene. The integration and expression of the mdr1 gene were observed indirectly by the expression of GFP, and fluorescence-activated cell sorter was used to evaluate the functional activity of permeability glycoprotein (P-gp) encoded by the mdr1 gene. The stimulating test was made in vitro to show pleiotropic drug resistance of transfected cells.</p><p><b>RESULTS</b>The isolated, cultured and expanded P-MSCs expressed stem cell markers such as CD29, CD44 and CD73, and showed osteogenic and adipogenic differentiation potentials under appropriate conditions. The expression of P-gp in the non-transfected P-MSCs cells was (0.4 +/- 0.1)%, but increased to (28.1 +/- 4.7)% after gene transfection (P < 0.01). And positive staining of P-gp located mainly at cell membrane and cytoplasm. Accumulation and extrusion assays showed that P-gp expressed by the transfected cells had pump-functional activity and could efflux daunomycin out of cells. The analysis of cell survival confirmed that transfected P-MSCs had a characteristic of multidrug resistance with a significant increase in the resistance to anticancer agents.</p><p><b>CONCLUSIONS</b>Transfer and expression of human mdr1 gene mediated by retrovirus vector conferred P-MSCs drug resistance. It might provide a new alternative to chemoprotection strategies.</p>

EEG Characteristics in Frequency Domain in Synaptic Dysfunction Rat Model / 现代生物医学进展

EEG characteristics in frequency domain were investigated in the frontal lobe, occipital lobe and hippocampus, i.e. cognition - related cortex of synaptic dysfunction model rat, providing electrophysiological basis for further study on plastic extent and nerve regeneration of the damaged neurons. Synaptic dysfunction model was made via microinjecting β - amyloid protein1 - 40 (Aβ1 - 40) into hippocampal CA1 area of rat. Morris water maze behavioral test was performed to evaluate the learning and memory function of model group. Then EEG in the above areas for two groups were recorded. The spectrum for two groups was performed and the characteristics in frequency domain were analyzed. The results showed: (1) The average escape latency in 3rd, 4th, 5th and 6th training times of model group are higher than those of normal. The average escape latency of normal group in 5th training time decreased more markedly than that in 2nd training time, while that of model group in 7th training time decreased more remarkably than that in 2nd training time (P ＜0.05). Without platform, the platform quadrant time percentage of model group was lower than the control (P ＜0.05). (2) Alpha rhythm in EEG of model rat was slowing down; alpha- band power decreased with peek frequency left shifted nearly 2Hz. And the power of delta - band and theta - band in frontal lobe, occipital and hippocampus all increased with different extent. The synaptic dysfunction model rats were made successfully by microinjecting Aβ1 - 40 method. The model rats show the decreased learning and memory dysfunction. EEG frequency spectrum features in model rat show slower alpha rhythm with power amplitude lower or loss, slow waves (delta and theta wave) increasing with higher power amplitude. These can be consistent with the EEG of Alzheimer's disease patients, which can provide electrophysiological basis for further plasticity and nerve regeneration study on the impaired cortex with synaptic dysfunction.

EEG Characteristics in Frequency Domain in Synaptic Dysfunction Rat Model / 现代生物医学进展

EEG characteristics in frequency domain were investigated in the frontal lobe, occipital lobe and hippocampus, i.e. cognition - related cortex of synaptic dysfunction model rat, providing electrophysiological basis for further study on plastic extent and nerve regeneration of the damaged neurons. Synaptic dysfunction model was made via microinjecting β - amyloid protein1 - 40 (Aβ1 - 40) into hippocampal CA1 area of rat. Morris water maze behavioral test was performed to evaluate the learning and memory function of model group. Then EEG in the above areas for two groups were recorded. The spectrum for two groups was performed and the characteristics in frequency domain were analyzed. The results showed: (1) The average escape latency in 3rd, 4th, 5th and 6th training times of model group are higher than those of normal. The average escape latency of normal group in 5th training time decreased more markedly than that in 2nd training time, while that of model group in 7th training time decreased more remarkably than that in 2nd training time (P ＜0.05). Without platform, the platform quadrant time percentage of model group was lower than the control (P ＜0.05). (2) Alpha rhythm in EEG of model rat was slowing down; alpha- band power decreased with peek frequency left shifted nearly 2Hz. And the power of delta - band and theta - band in frontal lobe, occipital and hippocampus all increased with different extent. The synaptic dysfunction model rats were made successfully by microinjecting Aβ1 - 40 method. The model rats show the decreased learning and memory dysfunction. EEG frequency spectrum features in model rat show slower alpha rhythm with power amplitude lower or loss, slow waves (delta and theta wave) increasing with higher power amplitude. These can be consistent with the EEG of Alzheimer's disease patients, which can provide electrophysiological basis for further plasticity and nerve regeneration study on the impaired cortex with synaptic dysfunction.