Methodology of Differentiation of Bone Marrow Cells into Megakaryocytes in Vitro / 中国实验血液学杂志

OBJECTIVE@#To explore the method for inducing the differentiation of bone marrow cells into megakaryocytes in vitro so as to use for evaluating the activity of traditional Chinese medicines.@*METHODS@#The bone marrow cells were separated from femurs and tibias of mice. The experiments were divided into 4 groups: control (no adding cytokines), TPO (adding 50 ng/ml TPO), TPO+SCF (50 ng/ml+50 ng/ml) and TPO+SCF+IL-6+IL-9 (50 ng/ml+50 ng/ml+20 ng/ml+20 ng/ml). The bone marrow cells in 4 groups were cultured in vitro for 6 d. Then the cell growth status was observed by the inverted microscopy, and the cell count was detected by using the automatic cell counter. The ratio and absolute count of megakaryocytes were detected by flow cytometry.@*RESULTS@#Compared with control, three induction methods could stimulate the differentiation of bone marrow cells into megakaryocytes in vitro. TPO could slightly enhance the differentiation of bone marrow cells into megakaryocytes. Both the combination of TPO and SCF, and the combination of TPO, SCF, IL-6 and IL-9 could intensively stimulate proliferation of bone morrow cells and promote the differentiation of bone marrow cells into megakaryocytes. The addition of IL-6 and IL-9 could decrease the proliferation of non-megakaryocytes, but promote the differentiation of bone marrow cells into megakaryocytes.@*CONCLUSION@#The optimized differentiation of bone marrow cells into megakaryocytes has been completed by co-induction regimen of TPO, SCF, IL-6 and IL-9, which can be used to screen and evaluate traditional Chinese medicines promoting formation of platelets.

Application of anoptomagnetic probe Gd-DO3A-EA-FITC in imaging and analyzing the brain interstitial space / 北京大学学报(医学版)

OBJECTIVE@#To investigate the application of the optical magnetic bimodal molecular probe Gd-DO3A-ethylthiouret-fluorescein isothiocyanate (Gd -DO3A-EA-FITC) in brain tissue imaging and brain interstitial space (ISS).@*METHODS@#In the study, 24 male SD rats were randomly divided into 3 groups, including magnetic probe group (n=6), optical probe group (n=6) and optical magnetic bimodal probe group (n=12), then the optical magnetic bimodal probe group was divided equally into magnetic probe subgroup (n=6) and optical probe subgroup (n=6). Referencing the brain stereotaxic atlas, the coronal globus pallidus as center level, the probes including gadolinium-diethylene triamine pentaacetic acid (Gd-DTPA), fluorescein isothiocyanate (FITC) and Gd-DO3A-EA-FITC of 2 μL (10 mmol/L) were injected into the caudate nucleus respectively, magnetic resonance imaging (MRI) was performed in the magnetic probe group and magnetic probe subgroup to image the dynamic diffusion and distribution of the probes in the brain ISS, a self-developed brain ISS image processing system was used to measure the diffusion coefficient, clearance, volume fraction and half-time in these two groups. Laser scanning confocal microscope (LSCM) was performed in vitro in the optical probe group and optical probe subgroup for fluorescence imaging at the time points 2 hours after the injection of the probe, and the distribution in the oblique sagittal slice was compared with the result of the first two groups.@*RESULTS@#For the magnetic probe group and magnetic probe subgroup, there were the same imaging results between the probes of Gd-DTPA and Gd-DO3A-EA-FITC. The diffusion parameters of Gd-DTPA and Gd-DO3A-EA-FITC were as follows: the average diffusion coefficients [(3.31±0.11)×10-4 mm2/s vs. (3.37±0.15)×10-4 mm2/s, t=0.942, P=0.360], the clearance [(3.04±0.37) mmol/L vs. (2.90±0.51) mmol/L, t=0.640, P=0.531], the volume fractions (17.18%±0.14% vs. 17.31%±0.15%, t=1.961, P=0.068), the half-time [(86.58±3.31) min vs. (84.61±2.38) min, t=1.412, P=0.177], the diffusion areas [(23.25±0.68) mm2 vs. (22.71±1.00) mm2, t=1.100, P=0.297]. The statistical analysis of each brain was made by t test, and the diffusion parameters were not statistically significant. Moreover, for the optical probe group and optical probe subgroup, the diffusion area of Gd-DO3A-EA-FITC [(22.61±1.16) mm2] was slightly larger than that of FITC [(22.10±1.29) mm2], the statistical analysis of each brain was made by t test, and the diffusion parameters were not statistically significant (t=0.713, P=0.492).@*CONCLUSION@#Gd-DO3A-EA-FITC shows the same imaging results as the traditional GD-DTPA, and it can be used in measuring brain ISS.

Dynamic visual analysis of neutrophils chemotaxis in peritoneal cavity of schizophrenic model in mice / 北京大学学报(医学版)

OBJECTIVE@#To compare the differences of neutrophils chemotaxis ability in peritoneal cavity between normal rats and schizopherenic rats with cell dynamic visualization system.@*METHODS@#In the study,18 healthy Kunming rats were randomly divided into 3 groups which were control group (n=6), 0.3 mg/kg MK-801 treatment group (n=6), 0.6 mg/kg dizocilpine maleate (MK-801) treatment group(n=6), extracted neutrophils separately, and observed the morphology and counted under a microscope. Each group of cells was divided into two parts for chemotactic experiment, called chemokine agent treatment group and no chemokine agent treatment group respectively, indicating control 1, 0.3 mg/kg MK-801 treatment 1,0.6 mg/kg MK-801 treatment 1 and control 2, 0.3 mg/kg MK-801 treatment 2,0.6 mg/kg MK-801 treatment 2. The dynamic migration of cells was recorded using the NIS-Elements software, and TAXIScan Analyzer 2 software was used to select 30 cells (n=30) in each group of cells and analyze cells migration trajectory, speed and distance, and use pair test and One-Way analysis of variance for statistical analysis.@*RESULTS@#The number of neutrophils in control group, 0.3 mg/kg MK-801 treatment group and 0.6 mg/kg MK-801 treatment group were(1.00±0.03)×104/mL,(0.05±0.02)×104/mL,(0.32±0.01)×104/mL respectively, the differences of results were statistically significant(P<0.05).Under the effect of chemotactic agent,the directional migration capability of neutrophils in control group 1, 0.3 mg/kg MK-801 treatment group 1 and 0.6 mg/kg MK-801 treatment group 1 were(0.85±0.11) radian,(1.00±0.11) radian,(0.96±0.10) radian respectively (P<0.05); the migration velocities of neutrophils were (0.09±0.02) μm/s,(0.12±0.01) μm/s,(0.14±0.01) μm/s respectively (P<0.05);the migration distances of neutrophils were (94.26±0.02) μm,(134.61±0.01) μm,(156.19±0.01) μm respectively(P<0.05).@*CONCLUSION@#Compared with neutrophils in peritoneal cavity of control group, the neutrophils in peritoneal cavity of schizophrenic rats have stronger chemotactic movement ability.

Imaging Gliomas with Nanoparticle-Labeled Stem Cells / 中华医学杂志(英文版)

<p><b>Objective:</b>Gliomas are the most common neoplasm of the central nervous system (CNS); however, traditional imaging techniques do not show the boundaries of tumors well. Some researchers have found a new therapeutic mode to combine nanoparticles, which are nanosized particles with various properties for specific therapeutic purposes, and stem cells for tracing gliomas. This review provides an introduction of the basic understanding and clinical applications of the combination of stem cells and nanoparticles as a contrast agent for glioma imaging.</p><p><b>Data Sources</b>Studies published in English up to and including 2017 were extracted from the PubMed database with the selected key words of "stem cell," "glioma," "nanoparticles," "MRI," "nuclear imaging," and "Fluorescence imaging."</p><p><b>Study Selection:</b>The selection of studies focused on both preclinical studies and basic studies of tracking glioma with nanoparticle-labeled stem cells.</p><p><b>Results:</b>Studies have demonstrated successful labeling of stem cells with multiple types of nanoparticles. These labeled stem cells efficiently migrated to gliomas of varies models and produced signals sensitively captured by different imaging modalities.</p><p><b>Conclusion</b>The use of nanoparticle-labeled stem cells is a promising imaging platform for the tracking and treatment of gliomas.</p>

Expression and significance of toll-like receptors 7 and 8 in brain and lung tissues of death cases caused by EV71 infection / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the significance of toll-like receptors (TLR) -7 and -8 in the pathogenesis of infection caused by Enterovirus type 71 (EV71) through measuring the expression of TLR7 and TLR8 in brain and lung tissues from the death cases caused by EV71 infection.</p><p><b>METHODS</b>Nine children who died of EV71 infection (EV71 group) were selected as study subjects, and 7 children who died of accidents or non-infectious diseases were used as the control group. Brain and lung tissues from the death cases in both groups at autopsy were collected, and immunohistochemistry was applied to detect the expression of TLR7 and TLR8 in lung and brain tissues in both groups. Integrated optical density (IOD) was applied for semi-quantitative analysis of the expression of TLR7 and TLR8.</p><p><b>RESULTS</b>Immunohistochemical results showed that the expression of TLR7 and TLR8 in lung and brain tissues was strongly positive in the EV71 group, and the IOD values in the EV71 group were also significantly higher than those in the control group (P<0.05). There was no significant difference in the expression of TLR7 and TLR8 between lung and brain tissues in the EV71 group (P>0.05).</p><p><b>CONCLUSIONS</b>TLR7 and TLR8 are highly expressed in lung and brain tissues from the patients who die of severe EV71 infection, suggesting that TLR7 and TLR8 may be involved in the pathogenesis of brain and lung damages caused by severe EV71 infection.</p>

Downregulation of cell-surface-expressed nucleolin inhibits the growth of hepatocellular carcinoma cells in vitro / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To detect the cell-surface-expressed nucleolin and investigate its tumor suppressing effect on the growth of hepatocellular carcinoma cells.</p><p><b>METHODS</b>To detect cell-surface-expressed nucleolin in the hepatocellular carcinoma cells by immunofluorescence and flow cytometry. To down-regulate the nucleolin expression level in hepatocellular carcinoma cells by RNA interference. The tumor-suppressing effect of cell-surface nucleolin on hepatocellular carcinoma cells was assessed by MTT and transwell chamber assays.</p><p><b>RESULTS</b>Nucleolin was expressed in the nuclei, cytoplasm and on the cell surface of hepatocellular carcinoma cells. ShRNA markedly decreased the nucleolin expression level in the cytoplasm and on the cell surface (P < 0.01), but the nuclear nucleolin remained unchanged. After downregulation of cell-surface nucleolin, MTT assays showed that the cell growth rate of hepatocellular carcinoma cells in the shRNA interference group was significantly inhibited as compared with that in the control group (P < 0.01). The transwell chamber assay showed that the mean transmembrane cell number in the shRNA interference group was significantly lower than that in the control group.</p><p><b>CONCLUSION</b>The results of this study show that downregulation of cell-surface nucleolin expression inhibits the growth of hepatocellular carcinoma cells in vitro.</p>

Suboccipital transtentorial approach to pineal region meningiomas by 3D-CTA assistance: surgical considerations / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the three-dimensional CT angiography (3D-CTA) assisted suboccipital transtentorial approach (Poppen's approach) in the treatment of pineal region meningioma.</p><p><b>METHODS</b>During the period of January 2005 to January 2010, 8 patients with pineal region meningioma were successfully treated using Poppen's approach through cerebral falx and tentorium. There were three male patients and five female patients were aged at a range of 41 - 64 years, average age was (54 ± 10) years. According to the Karnofsky performance scale (KPS), 5 patients' KPS scores were more than or equal to 80 and 3 were less than 80. MRI was used for the diagnosis of meningioma. 3D-CTA was applied to detect meningioma staining and blood supply. For preoperative concurrent hydrocephalus, follow-up observations were given. If hydrocephalus didn't get better or even became worse, ventriculoperitoneal shunt should be considered.</p><p><b>RESULTS</b>All the surgery were successfully performed, and venous complexes (VC) were well protected according to the CTA images. Out of the eight cases whose meningiomas were removed, one patient had got postoperative intracranial infection and recovered after given antibiotics. All patients were followed up for a period of 6 - 24 months. Preoperative concurrent hydrocephalus in 7 patients were improved. However, there was an aggravation of the hydrocephalus in one patient who was treated with ventriculoperitoneal shunt. The MRIs which were performed at the end of follow-up period, showed no recurrence of meningiomas, and preoperative symptoms were improved to varying degrees, 7 patients' KPS scores were more than or equal to 80 and 1 was less than 80. A χ(2) test was used to analyze and to make comparisons between preoperative and postoperative KPS. The significance was indicated (χ(2) = 1.33, P < 0.05).</p><p><b>CONCLUSIONS</b>For meningiomas in the pineal region, 3D-CTA is of great clinical value to distinguish the anatomic relationship among the meningioma, blood supply and VC. This case study has strongly supported using Poppen's approach assisted by 3D-CTA to proceed with the operation.</p>

Nanoparticles as a vaccine adjuvant of anti-idiotypic antibody against schistosomiasis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The development of new adjuvants for human use has been the focus of attention. This study's aim is to explore the possibility of using nanoparticle Ca nanoparticles (CA) as a vaccine adjuvant of anti-idiotypic antibody NP30 against schistosomiasis and its protective mechanisms.</p><p><b>METHODS</b>Nanoparticle CA-NP30 conjugate (CA-NP30) was fabricated. BALB/c mice were immunized actively with CA-NP30 to evaluate its effects of protective immunity on mice. The serum levels of specific IgG, IgG1 and IgG2a antibodies against NP30 and the concentrations of IFN-gamma and IL-4 in supernatant of splenocytes were determined via ELISA.</p><p><b>RESULTS</b>Nanoparticle CA could enhance significantly the protective immunity of NP30 against infection of Schistosoma japonicum and the worm reduction rose from 36.0% (NP30 alone) to 52.6%. The serum levels of specific IgG, IgG1 and IgG2a antibodies against NP30 increased remarkably, as compared with those of the group immunized with NP30 alone. The concentration of IFN-gamma in supernatant of splenocyte was drastically elevated [the groups immunized with CA-NP30 and NP30 alone were (493.80 +/- 400.74) pg/ml and (39.03 +/- 39.58) pg/ml, respectively], but the concentration of IL-4 showed no significant difference from that of NP30 alone [(27.94 +/- 9.84) pg/ml vs (27.28 +/- 14.44) pg/ml].</p><p><b>CONCLUSIONS</b>Nanoparticle CA could act as a vaccine adjuvant of anti-idiotypic antibody NP30 against schistosomiasis. The mechanism could be that CA-NP30 enhances humoral and cellular immune responses in mice.</p>