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OBJECTIVE@#To investigate the effect of low-intensity pulsed ultrasound (LIPUS) on hematopoietic function in rats after combined chemotherapy with doxorubicin and cyclophosphamide.@*METHODS@#Eighty rats were randomized into control group and LIPUS group (=40) for treatment with intraperitoneal injection of doxorubicin (2 mg/kg)+cyclophosphamide (20 mg/kg) for 4 consecutive days and continuous irradiation with LIPUS for 7 days following the injections, respectively. The white blood cells, red blood cells and platelets counts in each group were measured at 0, 4, 7, 9, 11, 14 and 18 days after the start of drug administration. The pathological sections of the bone marrow were examined at 0, 4 and 11 days, and the flow cytometry was performed for detecting the cell apoptosis; qPCR was performed for detecting the expressions of SCF, ICAM-1, and VCAM-1 mRNAs, and ELISA was used to detect the expressions of IL-3 and GM-CSF.@*RESULTS@#The white blood cell count was significantly higher in LIPUS group than in the control group ( < 0.05). Histopathological examination of the bone marrow revealed significantly increased hematopoietic tissue in LIPUS group ( < 0.05). Flow cytometry demonstrated an obviously lower cell apoptosis rate in the bone marrow in LIPUS group than in the control group ( < 0.05). Compared with those in the control group, the mRNA expression levels of ICAM-1 and VCAM-1 as well as the protein levels of IL-3 and GM-CSF were significantly increased in LIPUS group ( < 0.05).@*CONCLUSIONS@#LIPUS can alleviate the hematopoietic damage after combined chemotherapy with doxorubicin with cyclophosphamide probably by increasing the expressions of ICAM- 1, VCAM-1, IL- 3, and GM-CSF.
Subject(s)
Animals , Rats , Bone Marrow , Cyclophosphamide , Doxorubicin , Hematopoietic Stem Cell Transplantation , Ultrasonic WavesABSTRACT
Objective To discuss the clinical application value of microplate allochroic silica gel assay for rapidly detecting rifampicin(RIF)-resistance of Mycobacterium tuberculosis(MTB).Methods Fifty MTB clinical isolates preserved in the tuberculosis(TB) laboratory of Nantong Municipal Sixth People's Hospital were detected RIF-resistance by using the microplate allochroic silica gel assay and compared with the Bactec MGIT960 method.Then,the RIF susceptibility test in 40 clinical sputum smear-positive specimens were simultaneously detected by using the microplate allochroic silica gel assay and Bactec MGIT960.Finally the sensitivity,specificity and accuracy of the test results were compared.Results The optimal inoculation volume of MTB was 10-3 mg/mL,the optimal detection time was 7-10 d and the judging critical value of the RIF minimum inhibitory concentration(MIC) Was 1.00 μg/mL by microplate allochroic silica gel assay.With the Bactec MGIT960 test as the gold standard,the sensitivity,specificity and accuracies of microplate allochroic silica gel assay for RIF-resistance susceptibility test of smear-positive specimens were 94.12％,100％ and 97.37％ respectively.Conclusion Microplate Allochroic silica gel assay can be used for directly detecting the MTB sensitivity to RIF of in sputum specimens.
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Objective To investigate the impact of low intensity pulsed ultrasound (LIPUS) with different intensities on the migration of bone marrow mesenchymal stem cells (BMSCs) in vitro.Methods BMSCs were divided into control group,30 mW/cm2 group,60 mW/cm2 group and 90 mW/cm2 group.Control group was treated by sham LIPUS exposure,and the other three groups were treated by LIPUS with corresponding intensities.The impact of LIPUS on scratch healing was tested with scratch assay,and the interference of proliferation was eliminated with MTT assay.The migration of BMSCs were evaluated with transwell migration assay.The expression of F-actin was analyzed with fluorescein isothiocyanate (FITC) fluorescent coloration.Results 24 h and 48 h after LIPUS exposure,there were statistical differences of scratch area among groups (F=26.559,106.110,both P＜0.001),and the scratch area of control group was the largest ([0.93 ± 0.26)mm2 of 24 h after LIPUS exposure and [0.70 ± 0.11]mm2 of 48 h after LIPUS exposure),while that of 30 mW/cm2 group was the smallest ([0.47 ±0.21]mm2 of 24 h after LIPUS exposure and [0.19±0.10]mm2 of 48 h after LIPUS exposure).There was no statistical difference of scratch area among the four groups immediately after LIPUS exposure (F=2.921,P=0.063).MTT assay results showed there was no statistical difference of absorbance among the four groups immediately,nor 24 h,48 h after LIPUS exposure (F=1.616,0.720,1.408;P=0.196,0.544,0.378).Significant difference was found in the number of cells migrated through the transwell chamber among the four groups (F=43.145,P＜0.001),and the cell number of 30 mW/cm2 group was the largest (212.53±35.32),while that of the control group was the least (89.53±19.27).F-actin fluorescence staining results showed the morphology of F-actin was changed after LIPUS exposure.The cytoskeleton became narrow and elongated.Statistical difference of relative fluorescence intensity was found among the four groups (F 64.350,P＜0.001).The relative fluorescence intensity of 30 mW/cm2 group was the largest (125.43 ± 17.43),while that of control group was the least (51.94± 12.76).Conclusion LIPUS can promote the migration ability of BMSCs in vitro with the best intensity was 30 mW/cm2.
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Objective To establish a method of judge the pyrazinamide(PZA) susceptibility for Mycobacterium tuberculosis(MTB) with 24-hole micro-liquid culture silica gel color plate,and evaluate the clinical application value of this method.Methods According to the result of MGIT960,to detect PZA drug susceptibility for 30 MTB clinical isolates of whose PZA susceptibility were known with silica gel color plate.The effects of different pH value,different inoculation concentrations on the results were observed,and the optimum detection conditions were discussed.Finally,the 98 MTB clinical isolated of whose PZA susceptibility were unknown were simultaneously detected by gel color plate and MGIT960,the sensitivity,specificity and accuracy were judged by PZA drug sensitivity.Results The best pH was 5.8-5.9 and the best concentration was 2.5×10-1 mg/mL in gel color plate.The best results were read after 7-14 d.The sensitivity,specificity and accuracy were 95.50%,96.30%,and 98.21% respectively just at PZA critical concentration was 100 μg/mg.The sensitivity,specificity and accuracy were 90.90%,92.59%,and 91.84% respectively just at PZA critical concentration was 200 μg/mg.Conclusion The PZA susceptibility for MTB with 24-hole micro-liquid culture silica gel color plate is accurate,rapid,low-cost and simple.
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AIM:To establish a microthrombus model by carrageenan (Ca)/ lipopolysaccharides (LPS) intraperitoneal injection in rats with hyperhomocysteinemia (HHcy) and endothelial dysfunction induced by L-methionine intake. METHODS: ① Male Sprague Dawley rats were randomly divided into 2 groups: control and endothelial dysfunction (HHcy) groups. L-methionine was administered by gavage in HHcy group for total 4 weeks. Purified water was administered by gavage in control rats. Plasma Hcy, NO and vWF were examined and the thoracic aorta were excised after 4 weeks of L-methionine treatment to evaluate endothelial function. ② Male Sprague Dawley rats were randomly divided into 3 groups to establish a microthrombus formation model with Ca/ LPS: control, microthrombus formation (Ca/LPS) and endothelial dysfunction plus mitoarothrombus formation (HHcy+Ca/LPS) groups. Control rats were injected with normal saline (NS). Ca/LPS rats were intraperitoneally injected with carrageenan (Ca) and followed by lipopolysaccharides (LPS) 16 h later. HHcy+Ca/LPS rats were intragastric gavaged by L-methionine for total 4 weeks, and then were injected with Ca/LPS in the same way as Ca/LPS group. Cruor parameters and platelet count were detected at 20 h after LPS or NS injection and the mesentery microcirculation was monitored. Plasma NO and vWF were also detected at 24 h after LPS or NS injection. RESULTS: ① Plasma Hcy concentrations and vWF level were significantly increased in HHcy group, while plasma NO content was significantly decreased compared with that in control group. Endothelial dependent relaxation (EDR) of aortic rings was significantly decreased in HHcy group, suggesting endothelial damage/dysfunction was induced by HHcy. ② Mesentery capillary was obviously blocked by microthrombus in Ca/LPS rats and was blocked more seriously in HHcy+Ca/LPS rats. Cruor parameter results suggested that Ca/LPS rats were in hypercoagulable phase and HHcy+Ca/LPS rats were in hypocoagulable phase at 20 h after LPS injection. Platelet count and plasma NO content in HHcy+Ca/LPS group were significantly decreased, while plasma vWF level was significantly increased compared with Ca/LPS group. CONCLUSION: L-methionine intake induces severe HHcy and causes endothelial dysfunction in rats. Microcirculation dysfunction and microthrombosis can be caused by Ca/LPS intraperitoneal injection and may be aggravated by endothelial dysfunction.