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OBJECTIVE@#To explore the expression of Blimp1, ATF4 and CHOP in bone marrow mononuclear cells from patients with multiple myeloma as well as the effect of aspirin on their expression.@*METHODS@#Sixty untreated patients with multiple myeloma and 30 patients with relatively normal bone marrow were selected. Mononuclear cells from the bone marrow fluid were separated using Ficoll separation solution. CD138 plasma cells were sorted by immunomagnetic beads method. RT-PCR was used to detect the expression levels of Blimp1, ATF4 and CHOP mRNA in U266 cells cultured in vitro. The cells were divided into blank control group, negative control group (no-loaded virus transfection) and positive experimental group [LV-Blimp1-RNAi (40051-2) transfection] by lentivirus transfection. RT-PCR was used to detect the expression of Blimp1, ATF4 and CHOP mRNA in cells of different groups. U266 cells were stimulated in vitro with different concentrations of aspirin solution (0, 0.5 mmol/L, 2.5 mmol/L, 5.0 mmol/L) for 24, 48 h and 72 h, respectively. The ability of cell proliferation in different groups was measured by CCK-8. U266 cells were stimulated with different concentrations of aspirin for 48 hours. And the mRNA expression of Blimp1, ATF4 and CHOP was detected by RT-PCR.@*RESULTS@#Compared with plasma cells in normal group, the expression of Blimp1 mRNA in CD138 plasma cells of MM patients significantly increased (8.040±1.878), and the mRNA expression levels of ATF4 and CHOP significantly decreased (0.735±0.089; 0.837±0.062) (P＜0.05). U266 cells were cultured in vitro. Compared with the blank control group and the negative control group, the mRNA expression level of Blimp in the positive experimental group was significantly down-regulated after infection with LV-Blimp1-RNAi (40051-2) lentiviral expression vector (0.637±0.021). ATF4 and CHOP mRNA expression levels were significantly increased (1.452 ± 0.027; 1.721 ± 0.038) (P＜0.05). The proliferation of U266 cells decreased after stimulation with aspirin. In the range of (0.5-5) mmol/L, aspirin could significantly inhibit the proliferation of U266 cells. The inhibition effect of aspirin was increased along with prolongation of time and increase of concentrations. After aspirin stimulation of different concentrations for 48 hours, the expression level of Blimp1 in U266 cells decreased with increasing of drug concentration, while the expression levels of ATF4 and CHOP increased with increasing of drug concentration.@*CONCLUSION@#Inhibition of Blimp1 expression in multiple myeloma cells can promote the expression of ATF4 and CHOP. Aspirin can inhibit the proliferation activity of myeloma cells by down-regulating Blimp1 expression in myeloma cells and up-regulating ATF4 and CHOP expression, therefore plays an anti-tumor rote.
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A highly sensitive and selective bioluminescent probe for hydrazine (BPH) was designed, synthesized and evaluated for detection of hydrazine in vitro and in vivo. BPH was designed to include a specific recognition group (acetyl) of hydrazine at an appropriate modification site of the optical reporter hydroxyluciferin (D-luciferin), which showed excellent performance both in selectivity and sensitivity to hydrazine. The results showed that the bioluminescent probe BPH developed in this study is an innovative and widely applicable tool for detecting hydrazine in complex natural environment or in animals.
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BACKGROUND Angiostrongyliasis is caused by the nematode Angiostrongylus cantonensis and can lead to eosinophilic meningitis and meningoencephalitis in humans. The young adult worms play central pathogenic roles in the central nervous system (CNS); however, the underlying mechanism is unclear. Excretory-secretory products (ESPs) are good investigation targets for studying the relationship between a host and its parasite. OBJECTIVES We aimed to profile, identify, and characterise the proteins in the ESPs of A. cantonensis young adults. METHODS The ESPs of young adult worms were collected from culture medium after incubation ranging from 24 to 96 h. Proteomic and bioinformatics analyses were performed to characterise the ESPs. FINDINGS A total of 51 spots were identified, and the highly expressed proteins included two protein disulphide isomerases, one calreticulin, and three uncharacterised proteins. Subsequently, approximately 254 proteins were identified in the ESPs of A. cantonensis young adults via liquid chromatography-mass spectrometry (LC-MS/MS) analysis, and these were further classified according to their characteristics and biological functions. Finally, we identified the immunoreactive proteins from a reference map of ESPs from A. cantonensis young adults. Approximately eight proteins were identified, including a protein disulphide isomerase, a putative aspartic protease, annexin, and five uncharacterised proteins. The study established and identified protein reference maps for the ESPs of A. cantonensis young adults. MAIN CONCLUSIONS The identified proteins may be potential targets for the development of diagnostic or therapeutic agents for human angiostrongyliasis.
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Humans , Adolescent , Adult , Parasite Egg Count , Angiostrongylus cantonensis/parasitology , Feces/parasitologyABSTRACT
Objective To study the expression of zinc finger and BTB domain containing 20 (ZBTB20) in peripheral blood B cells of the patients with systemic lupus erythematosus (SLE),and to investigate its clinical significance in SLE.Methods The ZBTB20 mRNA expression level in peripheral blood CD19+B cells of 36 cases of SLE (SLE group) and 30 healthy controls(control group) was detected by RT-PCR.The ZBTB20 protein level was detected by western blot;the proportion of B cell subset was measured by flow cytometry.Then the relationship between the ZBTB20 mRNA expression with B cells subset proportion change and its correlation with clinical indicators[anti-dsDNA antibody,immunoglobulin G (IgG),anti-nucleosome antibody (ANA) and anti-extractable nuclear antigen (ENA) antibody] in SLE patients were analyzed.Results The expression level of ZBTB20 mRNA in peripheral blood CD19+B cells of the SLE group was significantly higher than that of the control group (P＜0.05).The peripheral blood ZBTB20 protein level was higher than that of the control group (P＜0.05);the peripheral blood B cells subsets and CD19+ B cells proportion in the SLE patients were decreased,while the CD19-CD138+ plasmocytes/CD19+ B cells ratio and CD19-CD138+ plasmocytes proportion were significantly increased (P＜0.05).The expression level of ZBTB20 mRNA in B cells of SLE patients was positively correlated with the ratio of CD19-CD138+ plasmocytes/CD19+ B cells (P＜0.05).The expression of ZBTB20 mRNA was positively correlated with anti-ds-DNA antibody,ANA and anti-ENA antibody (P＜0.05).Conclusion ZBTB20 might participate in the pathogenesis of SLE possibly via promoting B cells differentiation.
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OBJECTIVE:To observe clinical efficacy and safety of Mailuoning injection in adjunctive treatment of acute cere-bral infarction. METHODS:A total of 65 patients with acute cerebral infarction selected from neurology department of our hospital were divided into control group(32 cases)and observation group(33 cases)according to random number table. Control group was given conventional treatment. Observation group was additionally given Mailuoning injection 10 mL added into 0.9% sodium chlo-ride injection 250 mL intrnrenously,ivgtt,qd,on the basis of control group. Both group were treated for 15 d. Clinical efficacy as well as serum levels of ox-LDL,BNP and MMP-9,NIHSS score before and after treatment,the occurrence of ADR were com-pared between 2 groups. RESULTS:The response rate of observation group was 90.91%,which was significantly higher than 65.63%,with statistical significance(P0.05). After treatment,serum levels of ox-LDL,BNP and MMP-9,NIHSS score in 2 groups were all decreased significantly,and the observation group was significantly lower than the control group,with statis-tical significance (P0.05). CONCLU-SIONS:Mailuoning injection has significant therapeutic efficacy for acute cerebral infarction,can significantly reduce serum levels of ox-LDL,BNP and MMP-9,promotes neurological function and the recovery of patients with cerebral infarction with good safety.
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Ischemic stroke leads to high potentiality of mortality and disability.The current treatment for ischemic stroke is mainly focused on intravenous thrombolytic therapy.However,ischemia/reperfusion induces neuronal damage,which significantly influences the outcome of patients with ischemic stroke,and the exact mechanism implicated in ischemia/reperfusion injury remains unclear,although evidence shows that oxidative stress is likely to be involved.Betulinic acid is mainly known for its anti-tumor and anti-inflammatory activities.Our previous study showed that betulinic acid could decrease the reactive oxygen species (ROS) production by regulating the expression of NADPH oxidase.Thus,we hypothesized that betulinic acid may protect against brain ischemic injury in the animal model of stroke.Focal cerebral ischemia was achieved by using the standard intraluminal occlusion method and reperfusion enabled after 2 h ischemia.Neurological deficits were scored.Infarct size was determined with 2,3,5-triphenyltetrazolium chloride monohydrate (TTC) staining and the mRNA expression of NADPH oxidase 4 (NOX4) was determined by RT-PCR in infarct tissue.ROS generation and apoptosis in ischemic tissue were analyzed by measuring the oxidative conversion of cell permeable 2',7'-dichloro-fluorescein diacetate (DCF-DA) to fluorescent dichlorofluorescein (DCF) in fluorescence microplate reader and TUNEL assay,respectively.In Kunming mice,2 h of middle cerebral artery (MCA) occlusion followed by 24 or 72 h of reperfusion led to an enhanced NOX4 expression in the ischemic hemisphere.This was associated with elevated levels of ROS generation and neuronal apoptosis.Pre-treatment with betulinic acid (50 mg/kg/day for 7 days via gavage) prior to MCA occlusion prevented the ischemia/reperfusion-induced up-regulation of NOX4 and ROS production.In addition,treatment with betulinic acid could markedly blunt the ischemia/reperfusion-induced neuronal apoptosis.Finally,betulinic acid reduced infarct volume and ameliorated the neurological deficit in this stroke mouse model.Our results suggest that betulinic acid protects against cerebral ischemia/reperfusion injury in mice and the down-regulation of NOX4 may represent a mechanism contributing to this effect.
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<p><b>OBJECTIVE</b>To explore the changes in the threshold of auditory brainstem response (ABR) and [Ca(2+)]I and calmodulin (CaM) in cochlear nucleus of newborn mice infected by murine cytomegalovirus (MCMV) in the brain.</p><p><b>METHODS</b>Sixty-nine newborn mice were randomized into model group and control group. The model group (54 mice) was established by intracranial injection with MCMV viral suspension 20 l and the same volume of 0.9% sodium chloride was injected in the control group (15 mice). After 1 month, the ABR was tested in a sound-electric screen environment and the threshold was recorded. Then intracellular free calcium [Ca(2+)]i and the mRNA level of CaM in the cochlear nucleus were assayed by flow cytometry and RT-PCR.</p><p><b>RESULTS</b>Compare to the control group [(64.0 ± 1.3) dBSPL], the threshold of ABR in the model group [(84.5 ± 2.7) dBSPL] was increased (F = 2.789,P = 0.000). Moreover, in the model group the intracellular free calcium [Ca(2+)]i and the mRNA level of CaM in the cochlear nucleus were increased (F = 1.290, P = 0.000; F = 4.252, P = 0.023), and the differences were statistically significant.</p><p><b>CONCLUSIONS</b>The intracranial injection of MCMV can lead to abnormal changes in the threshold of ABR in mice, and the change of [Ca(2+) ]I/CaM in cochlear nucleus may be the important pathological basis of sensorineural hearing loss induced by MCMV infection.</p>