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Objective:To evaluate the value of improved pulmonary ultrasonography in the follow-up assessment of lung damage in patients who recovered from corona virus disease 2019(COVID-19).Methods:Twenty-two patients who were cured of COVID-19 in Quanzhou First Hospital from January to May 2020 were randomly selected and divided into 7 mild cases, 12 moderate cases and 3 severe cases according to the first high-resolution CT (HRCT) at admission. Six months after recovery, modified lung ultrasonography and HRCT were used prospectively to assess the lung damage and evaluate the correlation and consistency between the two techniques.Results:①There were significant differences in lung damage between the mild group and the moderate group, severe group (all P<0.05), while there was no significant difference between the moderate group and severe group ( P>0.05). ②There was good consistency between the improved lung ultrasound examination and HRCT (Kappa=0.776, P<0.001). ③There was a positive correlation between the score of improved pulmonary ultrasound examination and HRCT Warrick score ( r=0.755, P<0.001). Conclusions:Improved pulmonary ultrasonography can be used as a priority in the evaluation of pulmonary damage follow-up in patients with COVID-19 recovery, reducing the use of CT, and providing favorable evidence for further clinical management.
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Objective To investigate the value of fractional contrast medium bolus injection in improving the quality of CT portography.MethodsA total of 42 patients were randomly divided into two groups which were all given iohexol (350mgI/mL) as the contrast medium.20 patients in the group A were injected with conventional method (dosage of 100ml, rate of 4mL/s).The group B (22 patients) were treated with fractional contrast medium bolus injection, the first phase with 60mL contrast medium (rate of 4mL/s) and the second phase (10s delayed) with 40mL contrast medium (rate of 4m/s).The tube was washed by 20mL saline with the same rate of injection at both phases.The CT values and the image quality of the branches of the portal vein were evaluated according to the original and postprocessed images.Independent samples test was used to compare the CT values of the portal vein, splenic vein, superior mesenteric vein, hepatic parenchyma and portal vein-liver parenchyma.The subjective evaluation scores of the image quality were compared by wilcoxon.ResultsThe CT values of the portal vein, splenic vein and portal vein-liver parenchyma in the group B were significantly higher than that in the group A (t=3.317,3.523,P<0.01).There was no significant difference in CT values of hepatic parenchyma and superior mesenteric vein between the two groups.Subjective quality score in the group B was superior to that in the group A, and the difference was statistically significant.T The two evaluation physicians agreed well.ConclusionThe technique of fractional contrast medium bolus injection can significantly improve the image quality of CT portograghy.
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Objective To isolate and identify side population (SP) cells like cancer stem cell from human pancreatic cancer cell line SW1990, for the purpose of further evaluation of their biological characteristics. Methods Cell suspension was stained with Hoechst 33342 and PI. Then SP cells were analyzed in the fluorescence activated cell sorter. Cell growth viability was measured by MTT. Stem cell marker CD133 was determined by flow cytometry. Cloning forming efficiency was determined by cloning plating. Expression of ABCG2 protein was detected by Western blot analysis. Results The proportion of SP cells was 2.7%, however it could be completely blocked by verapamil. 9 days later, the value of A492 of SP cells was 2.1, the cloning forming efficiency was (38.7 ± 6.8) % , the positive rate of CD133 was 69.63%, which were significantly higher than cells 0. 5, ( 15.5 ± 2.8)%, 16.71% of corresponding non-SP( P <0.05). The expression of ABCG2 in SP cells was significantly higher than that in non-SP cells. Conclusions SP cells existed in human pancreatic cancer cells SW1990.
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AIM To investigate the protection of plicamycin on apoptosis in cerebellar granule neurons (CGN) of rat. METHODS TUNEL, Hoechst 33258 staining, agarose gel electrophoresis and fluorescein diacetate staining were used to detect morphological and biochemical characteristics of apoptosis in primary rat CGN. RESULTS Being pre-incubated with plicamycin for 1 h and lasting for 24 h, rat CGN apoptosis induced by low potassium basal modified Eagle′s medium for 24 h was inhibited in a plicamycin concentration-dependent manner. This effective concentrations of plicamycin were from 50 to 200 nmol·L-1, and the maximum inhibitory rate of plicamycin on CGN apoptosis was near 80% at 200 nmol·L-1. CONCLUSIONPlicamycin inhibits rat CGN apoptosis induced by low potassium.
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Aim To find out the relationship between muscarinic receptor and reactive oxygen species (ROS) and the probable differences between the four muscarinic receptor subtypes. Methods We transfected the plasmid encoding muscarinic receptor (including subtypes: M_1, M_2, M_3 and M_4) into PC12 cells. Then PC12 cells were exposed to hydrogen peroxide (H_2O_2), carbachol and other inhibitors such as atropine, LY294002 and PD98059. Results The results showed that activation of muscarinic receptor by carbachol protected PC12-M_1, PC12-M_2,PC12-M_3 and PC12-M_4 cells from apoptosis induced by H_2O_2. There was no statistical difference in the protective effect between these four muscarinic receptor subtypes. By using the inhibitors, we found that atropine and LY294002 blocked the protective effect of activation of muscarinic receptor on apoptosis induced by H_2O_2. Conclusion Activation of muscarinic receptor retarded the apoptosis induced by H_2O_2. There was no difference between the four muscarinic receptor subtypes. The protective effect was mainly mediated by the activation of muscarinic receptor and phosphatidylinositol-3 kinase (PI3K).
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AIM: To Screen and identify differentially expressed genes that involved in apoptosis model in rat cerebellar granule neurons (CGNs).METHODS: The rat cerebellar granule neurons were isolated and primarily cultured. Fluorescent differential display RT-PCR (FDD RT-PCR) was performed to screen differentially expressed ESTs in the apoptosis model of primarily cultured rat CGNs. ESTs were subcloned into pGEM-T EasyTM vector and then sequenced. Alignment assay in non-redunant database was applied for encoding information. Reverse Northern blotting was used to appraise the results from DDRT-PCR.RESULTS: 164 pieces of differentially expressed ESTs were obtained by FDDRT-PCR. 17 of them were subcloned and sequenced. 5 ESTs of 17 were confirmed to be positive results by reverse Northern blotting. CONCLUSION: DD-PCR is a rapid, simple-operation and sensitive method for screening differentially expressed genes, which would contribute to the molecular mechanisms of apoptosis/survive of CGNs.
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AIM: To analyze the subcellular localization of Arnt2 in rat cerebellar granule neurons (CGNs). METHODS: Based on the amino acids sequence of Arnt2 (LOCUS:NP_036913), the subcellular localization of Arnt2 in eukaryotic cells and the nuclear export signals (NES) of Arnt2 were predicted in CBS bioinformatics database. The subcellular localization of Arnt2 in rat cerebellar granule neurons was detected by the method of laser scanning confocal microscopy (LSM) analysis. RESULTS: It was predicted that Arnt2 located in nuclei of eukaryotic cells with the most probability, while located in cytoplasmic mitochondria with a slight possibility. A nuclear export signal was found in Arnt2 amino acids sequence, it was identified to be the leucine of No.143 that located in N-terminal of Arnt2 amino acids sequence. Finally, the result of LSM analysis shows nuclear localization of Arnt2 in rat CGNs. CONCLUSION: Arnt2 is located in nuclei of normal rat CGNs, it suggests that Arnt2 has the tendency to translocate into mitochondria after induced by some of inducible factors, for both the possibility of mitochondria localization and NES exist in Arnt2 amino acids sequence.
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AIM: To clone the full-length of 75A EST. METHODS: After the extraction of total RNA from primary cultured rat cerebellar granule cell of 7DIV in the medium containing 25 mmol/L KCl, T_4 DNA ligase-mediated 5' RACE was used to retrieve 5' unknown sequence of 75A EST, and the first round 5' RACE PCR product was subcloned into pGEM-T easy vector for sequence and homogeneous analysis. RESULTS: The first round of 5' RACE produce a 2.5 kb band, and 75A EST was identified to be partial sequence of Neuron-derived orphan receptor (Nor1) gene. After two more rounds RACE, we firstly cloned the full-length of Nor-1 cDNA. CONCLUSION: T_4 DNA ligase mediated 5' RACE is an efficient method to retrieve information about the 5' termini of mRNAs, and lay a foundation for further study which role Nor1 play in the cerebellar granule cell differentiation or survive.