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Objective To investigate the value of breast MRI-abbreviated protocol(BMRI-AP)compared with full field digital mammography(FFDM)and breast MRI-full diagnostic protocol(BMRI-FDP)in the diagnosis of early breast cancer with non-calcified.Methods A total of 95 cases patients with early breast cancer with non-calcified(the longest diameter of the lesion≤2 cm,regardless of the size of the carcinoma in situ)were retrospectively included.Clinical,pathological and imaging data of all patients were collected.All patients underwent FFDM and MRI scanning,and three examination regimens,including FFDM,BMRI-AP,BMRI-FDP,were further obtained.Classification was performed according to the breast imaging reporting and data system(BI-RADS)classification standard(fifth edition)developed by American College of Radiology(ACR),and pathological results were taken as the standard.The diagnostic efficacy for early breast cancer with non-calcified were compared among the different three imaging methods.Results The diagnostic accuracy of FFDM,BMRI-AP and BMRI-FDP for early breast cancer with non-calcified was 76.84%,93.68%and 95.79%,respectively,with statistically significant difference among three groups(χ2=20.558,P<0.001).The median(quartile distance)of BMRI-AP and BMRI-FDP scanning time were 478(5)s and 926(13)s,respectively,with statistically significant difference between the two groups(Z=-11.912,P<0.001).Conclusion The diagnostic accuracy of BMRI-AP is significantly better than that of FFDM and similar to that of BMRI-FDP for early breast cancer with non-calcified.In addition,BMRI-AP can significantly shorten the scanning time without reducing the diagnostic accuracy,which is expected to become a new breast cancer screening method.
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Objective:To explore the effect of NOD-like receptor thermal protein 3 ( NLRP3) knockout in γ-aminobutyric acid (GABA)-ergic neurons in the hippocampal CA1 area on improving cognitive dysfunction in mice after traumatic brain injury (TBI). Methods:Forty-eight healthy male NLRP3 flox/flox mice weighing 25-28 g were randomly divided into 4 groups ( n=12): sham-operated+control virus group (SV group), sham-operated+ NLRP3 specific knockout group (SG group), TBI+control virus group (TV group), TBI+ NLRP3 specific knockout group (TG group). TBI in the TV and TG groups was established by free-fall method, while surgical procedures such as scalp incision and cranial window opening without impact were given to the SV and SG groups. Adenovirus was injected into the hippocampal CA1 area of SG and TG groups 21 d before TBI to induce NLRP3 specific knockout in GABA-ergic neurons in the hippocampal CA1 area; empty virus was injected into the CA1 area of SV and TV groups. Cognitive function was evaluated using novel object recognition test 30 and 31 d after TBI, and learning and memory functions were assessed using Morris water maze test 32-36 d after TBI. Field potentials in the hippocampal CA1 area were recorded during novel object recognition 31 d after TBI. After behavioral tests, these mice were sacrificed. Immunofluorescent staining was used to detect the fluorescent intensity of microtubule-associated protein2 (MAP2), glutamic acid decarboxylase 67 (GAD67), and postsynaptic density protein 95 (PSD95) in the hippocampal CA1 area, as well as percentage of pyroptosis-associated inflammatory factor interleukin-18 (IL-18)/GAD67 double-positive neurons in total GAD67 positive neurons. Results:Compared with the SV and SG groups, the TV and TG groups had decreased novel object recognition index, decreased number of platform crossings during the experimental period, increased escape latency on day 3 and day 4 of the training period in Morris water maze test, decreased θ and γ oscillation power in the hippocampal CA1 area during novel object recognition, decreased fluorescent intensity of MAP2, GAD67, and PSD95 in the hippocampal CA1 area, increased percentage of IL-18/GAD67 double-positive neurons, with significant differences ( P<0.05). Compared with the TV group, the TG group had increased novel object recognition index, increased number of platform crossings in Morris water maze test, decreased escape latency during the training period, increased θ and γ oscillation power in the hippocampal CA1 area during novel object recognition, increased fluorescence intensity of MAP2, GAD67, and PSD95 in the hippocampal CA1 area, decreased percentage of IL-18/GAD67 double-positive neurons, with significant differences ( P<0.05). Conclusion:Specific inhibition of NLRP3 expression in GABA-ergic neurons in the hippocampal CA1 area can improve cognitive dysfunction in mice after TBI, whose mechanism may be related to inhibited GABA-ergic neuronal pyroptosis in the hippocampal CA1 area.
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Objective:To investigate the role and diagnostic value of miRNA-205 in chronic kidney disease (CKD) patients with vascular calcification.Methods:It was divided into in vitro cell experiment and retrospective cohort study. In vitro experiments were conducted by using rat thoracic aortic smooth muscle cells. Alizarin red staining and calcium content detection were used to detect the calcification of vascular smooth muscle cells (VSMCs). Alkaline phosphatase (ALP) test kit was used to measure ALP activity. Western blotting was used to detect the protein expression levels of osteogenic transcription factors runt-related transcription factor 2 (Runx2), α smooth muscle actin (α-SMA) and smooth muscle-22α (SM-22α) in VSMCs. qRT-PCR was used to detect miRNA-205 and Runx2 expression levels. The double luciferase reporter gene assay was used to verify the targeted relationship between miRNA-205 and Runx2. The non-dialysis patients with CKD 3-5 stage from June 2020 to January 2021 in the Department of Nephrology of Fourth Hospital, Hebei Medical University were selected. According to coronary artery calcium score (CACs), the patients were divided into non-calcification group (CACs=0), mild-moderate calcification group (0<CACs≤400), and severe calcification group (CACs > 400). Spearman correlation analysis was used to analyze the correlation between miRNA-205 and Runx2 and vascular calcification. Logistic regression model and receiver operating characteristic (ROC) curve analysis were used to analyze the ability of miRNA-205 to predict the vascular calcification in patients with CKD. Results:(1)Compared with the control group, calcium nodules were more, and the calcium content, ALP activity and Runx2 protein level were higher, and the expression levels of miRNA-205, α-SMA and SM-22α were significantly lower in high phosphorus group (all P<0.05). Overexpression of miRNA-205 significantly reduced the calcification of VSMCs and Runx2 protein level, and increased the protein levels of α-SMA and SM-22α (all P<0.05). miRNA-205-5p reduced the activity of luciferase in the wild-type Runx2-3'-end non-coding region plasmid. (2) Eighty CKD patients were enrolled, with age of (57.50±14.93) years old and 49 males (61.3%). The results of comparison of miRNA-205 and Runx2 expression levels in non-calcification group ( n=26), mild- moderate calcification group ( n=30) and severe calcification group ( n=24) showed that, the higher degree of calcification, the lower miRNA-205 expression level and the higher Runx2 mRNA expression level (all P<0.05). miRNA-205 was negatively correlated with CACs ( r=-0.50, P<0.01) and Runx2 was positively correlated with CACs ( r=0.55, P<0.01). Multivariate logistic regression analysis results suggested that miRNA-205 ( OR=0.451, 95% CI 0.122-0.873) was an independent influencing factor of vascular calcification in CKD patients. The area under the ROC curve of miRNA-205 and miRNA-205 combined with Runx2 for predicting vascular calcification were 0.796 (95% CI 0.697-0.859) and 0.924 (95% CI 0.866-0.982), respectively. Conclusions:miRNA-205 inhibits vascular calcification by targeting Runx2 to negatively regulate osteogenetic phenotype transformation of VSMCs and is expected to be an early diagnostic marker of vascular calcification in CKD patients.
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Objective:To evaluate the role of astrocytic NOD-like receptor protein 3 (NLRP3) in the lateral hypothalamus (LHA) in anxiety-like behaviors after hemorrhagic shock and resuscitation in mice.Methods:Forty-eight clean-grade male C57BL/6 mice, aged 10 weeks, weighing 25-30 g, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (group C), hemorrhagic shock and resuscitation group (group H), hemorrhagic shock and resuscitation + adeno-associated virus group (group HI), and hemorrhagic shock and resuscitation + control virus group (group HIV). The model of hemorrhagic shock and resuscitation was developed by bleeding and re-transfusion through the femoral vein in H, HI and HIV groups. At 21 days before developing the model, AAV-GfaABC1D-EGFP-Cre was injected into bilateral LHA in group HI, and AAV-GfaABC1D-EGFP was administered as a control in group HIV. Anxiety-like behaviors were evaluated by EPM-maze and bead-burying tests at 14 days after resuscitation. Mice were immediately sacrificed at the end of behavioral tests, and LHA-containing brain tissues were obtained for determination of co-localization of NLRP3 with glial fibrillary acidic protein (GFAP), the fluorescence intensity of Wisteria floribunda agglutinin was measured using immunofluorescent staining to reflect the expression of extracellular matrix in the LHA, and the percentage of cleaved caspase-1/GFAP and IL-18/GFAP positive cells in total cells was calculated. Results:Compared with group C, the number of buried beads and percentage of time of staying at the open arm were significantly decreased, the expression of extracellular matrix in the LHA was down-regulated, and the percentage of cleaved caspase-1/GFAP and IL-18/GFAP positive cells was increased in H, HI and HIV groups, and the co-localization coefficient of NLRP3 and GFAP was significantly decreased in group HI ( P<0.01). Compared with group H, the number of buried particles and percentage of time of staying at the open arm were significantly decreased, the expression of extracellular matrix in the LHA was up-regulated, the co-location coefficient of NLRP3 and GFAP was decreased, the percentage of cleaved caspase-1/GFAP and IL-18/GFAP positive cells was decreased ( P<0.01), and no significant change was found in the parameters mentioned above in group HIV ( P>0.05). Conclusions:Anxiety-like behaviors after hemorrhagic shock and resuscitation is associated with astrocytic NLRP3-induced pyroptosis in the LHA and reduction of extracellular matrix in mice.
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@#Bacterial antimicrobial resistance (AMR) is a globally serious problem that threatens public health security.Misuse and abuse of antibiotics cannot achieve the effect of treating bacterial infectious diseases, but will trigger the SOS response of bacteria, exacerbating the evolution of bacterial AMR and the spread of resistant bacteria.This article focuses on antibiotic-resistant bacteria, briefly introduces the pathogenesis of bacterial AMR and SOS response, and systematically summarizes the determination and mechanism study of bacterial AMR based on microfluidics and mass spectrometry.This article provides theoretical basis for AMR-related drug target mining and new drug development, aiming to develop new methods for rapid detection of bacterial AMR and new methods for bacteria inhibition, and promote the diagnosis and treatment of clinical bacteria infectious diseases.
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Objective:To investigate the effect of Spautin-1 (an inhibitor of autophagy) on improving anxiety-like behaviors and its mechanism in mice after traumatic brain injury (TBI).Methods:Thirty-six C57BL/6 mice were randomly divided into sham-operated group, TBI group, and TBI+Spautin-1 group ( n=12); TBI models in the latter two groups were established by modified Feeney free fall epidural impingement method. Mice in TBI+Spautin-1 group were administered with Spautin-1 (2 μL, 10 mmol/L) into the lateral ventricle 10 min after modeling, but mice in the other two groups were only injected with same volume of solvent. Neuropathy Symptom Score (NSS) was used to evaluate the functions of motor, sensory and reflexes of mice on 1 st, 7 th and 14 th d of modeling. On 15 th and 16 th d of modeling, open field test and elevated plus maze test were used to evaluate the anxiety-like behaviors in mice. The number of Nissl bodies in the amygdala of mice was calculated by Nissl staining 16 d after modeling. The numbers of neuron specific nucleoprotein (NeuN) positive cells, interleukin (IL)-18 and IL-1β positive astrocytes in the amygdala were detected by immunofluorescent staining. Western blotting was used to detect the autophagy-and pyrotopic-associated protein expressions in the amygdala region of mice. Results:(1) As compared with the sham-operated group, the TBI group and TBI+Spautin-1 group had significantly increased NSS scores on 1 st and 7 th d of modeling ( P<0.05). (2) Open field test showed that as compared with the sham-operated group, the TBI group and TBI+Spautin-1 group had significantly smaller number of crossing grids, significantly decreased percentage of time spending in the central zone ([central area residence time/total time] × 100%), significantly decreased percentage of frequencies entering into opening arm (OE) (OE/[OE+frequencies of entering closing arm]×100%) and opening arm time (OT) percentage (OT/[OT+time of closing arm]×100%); as compared with the TBI group, TBI+Spautin-1 group had significantly larger number of crossing grids, and significantly increased time percentage spending in the central zone, OE percentage, and OT percentage ( P<0.05). (3) As compared with sham-operated group, the TBI group and TBI+Spautin-1 group had significantly smaller numbers of Nissl bodies and NeuN positive cells in the amygdala of mice ( P<0.05); as compared with TBI group, TBI+Spautin-1 group had significantly larger numbers of Nissl bodies and NeuN positive cells in the amygdala of mice ( P<0.05). (4) As compared with the sham-operated group, the TBI group and TBI+Spautin-1 group had significantly increased percentages of IL-1β and IL-18 positive astrocytes in amygdala of mice ( P<0.05); as compared with the TBI group, the TBI+Spautin-1 group had significantly decreased percentages of IL-1β and IL-18 positive astrocytes in amygdala of mice ( P<0.05). (5) As compared with sham-operated group, the TBI group and TBI+Spautin-1 group had significantly higher protein expressions of NOD-like receptor family protein 3 (NLRP3), activated cysteine aspartate protease-1 (Caspase-1), pore-forming protein D-N terminal fragment (GSDMD-N), ubiquitin specific peptidase (USP) 13 and B-lymphocytoma-2 interacting protein (Beclin1), and statistically higher ratio of microtubule-associated protein 1 light chain (LC)3 II/LC3 I ( P<0.05); as compared with TBI group, the TBI+Spautin-1 group had significantly decreased protein expressions of NLRP3, activated Caspase-1, GSDMD-N, USP13 and Beclin1 in the amygdala, and statistically lower ratio of LC3 II/LC3 I ( P<0.05). Conclusion:Spautin-1 improves the anxiety-like behaviors in mice after TBI, whose mechanism may be associated with the inhibition of astrocytic pyroptosis in the amygdala.
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Objective:Our previous micro-array study revealed that long non-coding RNA (lncRNA), such as ENST00000538790 and NR_125790, were differently expressed in parathyroid carcinoma compared with those in parathyroid adenoma. Diagnostic value of lncRNAs (ENST00000538790 and NR_125790) and scoring models derived from these lncRNAs in parathyroid carcinoma were investigated in this study.Methods:Fifty-seven fresh tissue samples from patients with hyperparathyroidism were collected. Eleven patients were diagnosed with parathyroid carcinoma, while 46 patients were found with parathyroid adenoma. The expression levels of five lncRNAs (ENST00000511928, ENST00000538790, ENST00000618339, NR_125790, and ENST00000485384) were detected with realtime quantitative PCR (RT-qPCR). LncRNA scores were calculated using these lncRNAs, and their value in parathyroid carcinoma diagnosis were also assessed.Results:It was found that expression levels of ENST00000511928 and ENST00000538790 were up-regulated in parathyroid carcinoma compared with parathyroid adenoma ( P<0.05), while the levels of ENST00000618339, NR_125790, and ENST00000485384 were decreased in parathyroid carcinoma ( P<0.05). Among these lncRNAs, ENST00000538790 and NR_125790 were independent risk factors for parathyroid carcinoma ( P<0.05). Hence, "log score" and "logistic score" were calculated with ENST00000538790 and NR_125790 levels. The areas under the receiver operating characteristic curve for "log score" and "logistic score" were up to 0.935 and 0.943 respectively in parathyroid carcinoma ( P<0.05), and were found to be greater than those from single lncRNA or classic clinical indices, such as serum calcium ( P<0.05). Conclusion:LncRNA scores with lncRNA ENST00000538790 and NR_125790 may play potential role in diagnosis of parathyroid carcinoma.
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Helicobacter pylori (Hp) is a Gram-negative bacterium selectively colonized in gastric mucosa, and is one of the main factors inducing chronic gastritis and even gastric cancer. Recent studies have shown that Hp infection induces various stress responses, including oxidative stress, nitrosative stress and endoplasmic reticulum stress. Hp can stimulate neutrophils, macrophages and gastric epithelial cells to express reactive oxygen species and reactive nitrogen species, resulting in excessive accumulation of reactive oxygen species and reactive nitrogen species, aggravating inflammation and damage to gastric mucosa. Such long-term inflammation and oxidative stress may increase the risk of cancer. In addition, Hp induced endoplasmic reticulum stress plays an important role in the early stages of precancerous lesion formation. This article reviewed the research progress of role of stress response induced by Hp infection in gastric mucosal diseases.
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Objective:To evaluate the value of synthetic MRI combined with DWI in the diagnosis of benign and malignant breast lesions.Methods:The data of 184 consecutive patients with suspected breast lesions in Yunnan Cancer Hospital from July to September 2019 were prospectively analyzed. All patients were randomly assigned to training group ( n=110) and validation group ( n=74), and underwent conventional MRI and synthetic MRI respectively before and after contrast injection. At the maximum slice of the lesion, the ROI was drawn along the edge and recorded as "tumor". In the solid area with the most obvious tumor enhancement, the second ROI was drawn and recorded as "local". At the same time, ADC values (ADC local and ADC tumor) and relaxation time values (T local and T tumor) were measured. T and T + represented the relaxation time value of the ROI pre-and post-contrast scanning. ΔT% represented the relative change rate in T value between pre-and post-contrast scanning.The rank sum test was used to test the quantitative parameters of benign and malignant breast lesions in the training group and the validation group, and the variables with P<0.05 were included in the binary logistic regression analysis to screen the independent variables and establish the prediction model. The area under ROC curve was used to evaluate the discrimination of parameters and models. The clinical applicability of model was analyzed by decision curve analysis (DCA). Results:In the training group, univariate analysis showed that there were significant differences in T 1tumor, T 1+tumor, ΔT 1% tumor, T 2local, T 2+local, T 2tumor and T 2+tumor, ADC local, ADC tumor between benign and malignant breast lesions ( P<0.05). Multivariate logistic regression analysis showed that T 1+tumor, ΔT 1% tumor, T 2tumor, ADC local, ADC tumor were independent variables in the diagnosis of breast cancer. The relaxation time model (model A: T 1+tumor, ΔT 1% tumor, T 2tumor) and ADC model (model B: ADC local, ADC tumor) established by combining the above variables had the same diagnostic efficiency (AUC=0.905, 0.914, Z=-1.874, P=0.062), and the multi-parameter combination model (model C: T 1+tumor, ΔT 1% tumor, T 2tumor, ADC local, ADC tumor) had the highest diagnostic efficiency (AUC=0.965). DCA analysis showed that when the threshold probability ranges between 21%-99% (training cohort) and 15%-99% (validation cohort), the net benefit of model C was better than model A and B. Conclusion:The multi-parameter combined prediction model established based on the relaxation time value and ADC can identify breast cancer efficiently and can be used as an auxiliary diagnostic tool.
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Objective:To evaluate the effect of selective inhibitor of caspase-1 VX-765 on cognitive function in a rat model of hemorrhage shock and resuscitation (HSR).Methods:Forty-eight clean-grade healthy male Sprague-Dawley rats, aged 9-10 weeks, weighing 350-400 g, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (S group), HSR group (H group), VX-765 group (V group), and solvent control group (C group). The rats in H, V and C groups were subjected to hemorrhage by bleeding from femoral vein to achieve mean arterial pressure of 25-35 mmHg which was maintained at this level for 60 min followed by resuscitation with shed blood within 15 min to restore blood pressure, and normal saline was infused when needed.VX-765 1 mg/kg and 0.4% polyethylene glycol 1 mg/kg were intravenously injected via the femoral vein immediately after the end of resuscitation in V and C groups, respectively.Six rats in each group were selected and sacrificed at 12 h after the end of resuscitation, and the cerebral cortex was removed for determination of neuronal pyroptosis (by immunofluorescence) and degree of cortical edema (using T2-weighted imaging). Cognitive function was measured by open field test on day 7 after resuscitation in the rest 6 rats in each group. Results:Compared with S group, the pyroptosis rate in cortical neurons at 12 h after resuscitation and degree of cortical edema were significantly increased, the distance in the central square and the number of standing on the back legs were decreased on day 7 after resuscitation, and the time spent in the central square was shortened in H, V and C groups ( P<0.05). Compared with H and C groups, the pyroptosis rate in cortical neurons at 12 h after resuscitation and degree of cortical edema were significantly decreased, the distance in the central square and the number of standing on the back legs were increased on day 7 after resuscitation, and the time spent in the central square was prolonged in V group ( P<0.05). Conclusion:VX-765 can improve the cognitive function, and the mechanism may be associated with inhibiting pyroptosis in cortical neurons in a rat model of HSR.
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Objective:To evaluate the effect of VX-765 on cognitive function in acute rapid eye movement (REM) sleep-deprived juvenile rats.Methods:Thirty-six clean-grade healthy male Sprague-Dawley rats, aged 3-4 weeks, weighing 52-101 g, were divided into 3 groups ( n=12 each) using a random number table method: control group (group C), acute REM group (group AREM) and VX-765 group (group V). Sleep deprivation model was established by modified multi-platform water environment method.In group V, VX-765 solution 10 mg/kg was intravenously injected via the tail vein at 9: 00 a. m.every day for 4 consecutive days.The equal volume of normal saline was given instead in C and AREM groups.Morris water maze and novel object recognition tests were performed for 4 consecutive days during sleep deprivation.The rats were then sacrificed after the end of Morris water maze and novel object recognition tests on 5th day, and hippocampi were removed for determination of the expression of interleukin-1beta (IL-1β) and IL-18 by Western blot. Results:Compared with group C, the latency of novel object recognition was significantly prolonged, the percentage of novel object exploration was shortened, and the number of head exploration was decreased, the percentage of novel object exploration and discrimination index were decreased, the number of crossing the original platform in Morris water maze test was reduced, the time of staying at the target quadrant was shortened, and the expression of IL-1β and IL-18 was up-regulated in AREM and V groups ( P<0.05). Compared with group AREME, the latency of novel object recognition was significantly shortened, the percentage of novel object exploration was prolonged, and the number of head exploration was increased, the percentage of novel object exploration and discrimination index were increased, the number of crossing the original platform in Morris water maze test was increased, the time of staying at the target quadrant was prolonged, and the expression of IL-1β and IL-18 was down-regulated ( P<0.05). Conclusion:VX-765 can improve the cognitive function in acute REM sleep-deprived juvenile rats, which is related to inhibiting hippocampal inflammatory responses.
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Objective:To explore the role and mechanism of lysine methyltransferase SET8 in calcification induced by high phosphorus in vascular smooth muscle cells (VSMCs).Methods:(1) Male SD rats were selected for in vivo experiments and randomly divided into sham operation group and chronic renal failure vascular calcification group. The thoracic aorta was taken and calcification was detected by von Kossa staining. The expression of SET8 and Caspase-3 was detected by immunohistochemistry. (2) VSMCs were randomly divided into normal group and high phosphorus group (10 mmol/L β-glycerophosphate). Cellular calcification was detected by O-cresol hydrazide complex colorimetric assay and alizarin red staining. Apoptosis was detected by flow cytometry. The expressions of SET8, AKT and Caspase-3 were detected by RT-PCR and Western blotting. (3) In order to further verify the role of SET8 in the apoptosis of VSMCs, liposome transfection was used, and cells were divided into three groups: SET8-shRNA group, empty plasmid group and normal control group. Cellular calcification was detected by O-cresol hydrazide complex colorimetric assay and alizarin red staining. Apoptosis was detected by flow cytometry. The expressions of SET8, AKT and Caspase-3 were detected by RT-PCR and Western blotting. Results:(1) In vivo experiments, compared with the sham operation group, vascular calcium deposition in the chronic renal failure vascular calcification group was significantly increased ( P<0.05). Immunohistochemistry results showed that SET8 expression was significantly decreased and Caspase-3 was significantly increased in the vascular calcification group (both P<0.05). Correlation analysis showed that SET8 was negatively correlated with vascular calcification and Caspase-3 ( r=-0.948, P<0.01; r=-0.961, P<0.01). (2) In vitro, the calcium deposition in the high-phosphorus group was significantly higher than that in the normal group ( P<0.05). The results of flow cytometry showed that the number of apoptosis in the high-phosphorus group was significantly higher than that in the normal group ( P<0.05). RT-PCR and Western blotting showed that, compared with the normal group, the mRNA and protein expression of SET8 and AKT in the high-phosphorus group decreased, and the mRNA and protein expression of Caspase-3 increased (all P<0.05). (3) After interference of SET8 gene expression, calcification and apoptosis of VSMCs significantly increased, AKT mRNA and protein expression decreased, and Caspase-3 mRNA and protein expression increased (all P<0.05). Conclusions:SET8 can inhibit vascular calcification. One of the possible mechanisms is to inhibit the expression of Caspase-3 via promoting AKT activation, thereby inhibiting the apoptosis of VSMCs, and then participating in the regulation of VSMCs calcification induced by high phosphorus.
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Objective To explore the role of CORM-3 in alleviating cognitive dysfunction and cortical neuronal pyroptosis of rats exposed to hemorrhage shock and resuscitation. Methods One hundred and sixty-eight male SD rats, weighting 350-400 g, in accordance with random number table, were divided into 4 groups (n=42):sham-operated group, hemorrhage shock and resuscitation (H group), hemorrhage shock and resuscitation plus CORM-3 (CO group), hemorrhage shock and resuscitation plus iCORM-3 (ICO group). The rat hemorrhagic shock resuscitation models were established in H, CO and ICO groups:bleeding from femoral vein was performed to achieve mean arterial pressure of 25-35 mmHg (1 mmHg=0.133 kPa) for 60 min;and then, the collected blood was returned to the body within 15 min to reach the initial blood pressure level as resuscitation, and normal saline was injected if necessary. The rats in CO group were injected CORM-3 (4 mg/kg) via femoral vein after resuscitation, the rats in ICO group were injected iCORM-3 (4 mg/kg), and rats in the sham-operate group and H group were only injected equal amount of normal saline containing DMSO. Rats were sacrificed for cortex 12 h after end of resuscitation;CO content was assessed by gas chromatograph assay; Western blotting was used to detect the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and BTB-CNC homology 1 (Bach1) in the nuclear, and heme oxygenase-1 (HO-1), interleukin (IL)-1β and IL-18 in cytosol; and neuronal pyroptosis rate was detected by cleaved caspase-1-Cy3/neuron-specific nucleoprotein (NeuN)-FITC/DAPI. Thirty d after resuscitation, open field test was used to assess the cognitive ability of the rats. Results At 12 h after resuscitation, as compared with sham-operated group, rats in the H, CO and ICO groups had significantly increased CO content, neuronal pyroptosis, ratio of Nrf2/Bach1, and expressions of HO-1, IL-1β and IL-18, statistically longer time spending in the central square, significantly smaller times crossing the grid and times standing on the back legs (P<0.05). As compared with H and ICO group, CO group had significantly increased CO content, ratio of Nrf2/Bach1, and HO-1 expression, times crossing the grid, times standing on the back legs, but significantly decreased neuronal pyroptosis, expressions of IL-1β and IL-18, and time spending in the central square (P<0.05). Conclusion CORM-3 can reduce the neuronal pyroptosis rate and alleviate cognitive dysfunction in rats with hemorrhagic shock and resuscitation, whose mechanism may be related to the increase of HO-1 expression after up-regulation of Nrf2/Bach1 ratio.
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Objective To evaluate the effect of carbon monoxide (CO) postconditioning on pyroptosis induced by oxygen-glucose deprivation and restoration (OGD/R) in rat hippocampai neurons and the relationship with mitochondrial permeability transition pore (mPTP)/reactive oxygen species (ROS) signaling pathway.Methods Primary hippocampal neurons were cultured in vitro,seed in 6-well or 96-well plates,and divided into 5 groups (n =24 each) using a random number table method:control group (C group),OGD/R group,CO postconditioning group (CO group),specific mPTP opener atractyloside plus CO postconditioning group (ACO group),and specific ROS inducer antimycin A plus CO postconditioning group (KCO group).Neurons were subjected to O2-glucose deprivation (OGD) for 16 h followed by restoration of O2-glucose supply for 24 h to establish the model of OGD/R injury.In group CO,neurons were exposed to 2％ CO-5％ CO2 for 3 h at 37 ℃ starting from the end of OGD,followed by normal culture for 21 h.In ACO and KCO groups,atractyloside 20 μmol/L and antimycin A 50 μmol/L were added at the end of OGD,respectively,and the other treatments were similar to those previously described in group CO.Neuronal pyroptosis rate was determined using double immunofluorescent staining cleaved caspase-1-AlexaFluor 568/DAPI after the end of treatments in each group.The neuronal survival rate was determined by MTT,opening of mPTP by Calcein-AM fluorescence,ROS content by DCFH-DA,and expression of interleukin1beta (IL-1β) and IL-18 by Western blot.Results Compared with C group,neuronal pyroptosis rate,ROS content and opening of mPTP were significantly increased,the neuronal survival rate was decreased,and the expression of IL-1β and IL-18 was up-regulated in the other groups (P＜0.05).Compared with OGD/R group,neuronal pyroptosis rate,ROS content and opening of mPTP were significantly decreased,the neuronal survival rate was increased,and the expression of IL-1β and IL-18 was down-regulated in CO,ACO and KCO groups (P＜0.05).Compared with CO group,neuronal pyroptosis rate and ROS content were significantly increased,the neuronal survival rate was decreased,and the expression of IL-1β and IL-18 was up-regulated in ACO and KCO groups,and opening of mPTP was significantly inctreased in ACO group (P＜0.05).Conclusion CO postconditioning can inhibit OGD/R-induced pyroptosis in rat hippocampal neurons,and the mechanism is related to inhibiting mPTP/ROS signaling pathway.
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Objective To evaluate the role of hippocampal extracellular signal-regulated kinase 1/2 (ERK1/2) in exogenous carbon monoxide (CO)-induced improvement of cognitive function in a rat model of hemorrhage shock and resuscitation.Methods Ninety clean-grade healthy male Sprague-Dawley rats,aged 9-10 weeks,weighing 350-400 g,were divided into 5 groups (n=18 each) using a random number table method:sham operation group (S group),hemorrhage shock and resuscitation group (H group),CO group,PD98059-CO group (PCO group) and PD98059 group (P group).Hemorrhagic shock was induced by withdrawing blood from the femoral vein until mean arterial pressure was reduced to 25-35 mmHg which was maintained for 60 min and resuscitated by infusing the blood withdrawn over 15 min until the initial blood pressure was achieved,and normal saline was infused when needed.Rats were exposed to air mixture containing 1％ CO for 3 h in a glass box after the end of resuscitation in group CO.ERK1/2 inhibitor PD98059 (30 μmol/L) 30 μl was injected into the cerebral ventricle at 30 min before hemorrhage in PCO and PC groups.Right femoral artery and vein were only cannulated,and normal saline was injected into the cerebral ventricle in group S.Rats were sacrificed at 3 h after the end of resuscitation,brains were removed and hippocampi were isolated for determination of CO content (by gas chromatograph assay).Cognitive function was assessed by Morris water maze test at 15 days after the end of resuscitation,rats were then sacrificed and hippocampi were isolated for determination of cell apoptosis in hippocampal CA1 region by TUNEL and cleaved caspase-3 immunofluorescence,and the apoptosis rate was calculated.Rats were sacrificed at 6 h after the end of resuscitation,and hippocampi were isolated to detect the expression of phosphorylated ERK1/2 (p-ERK1/2),Bcl-2 and Bax by Western blot.Results Compared with group S,the escape latency was significantly prolonged in H,PCO and P groups,and the hippocampal CO content and apoptosis rate were increased,the expression of cleaved caspase-3 and ERK1/2 was up-regulated,and the Bcl-2/Bax ratio was decreased in H,CO,PCO and P groups (P＜0.05).Compared with group H,the escape latency was significantly shortened,the hippocampal CO content was increased,and the apoptosis rate was decreased,the cleaved caspase-3 expression was down-regulated,p-ERK1/2 expression was upregulated,and Bcl-2/Bax ratio was increased in group CO,and the escape latency was significantly prolonged,the hippocampal CO content was decreased,the apoptosis rate was increased,the cleaved caspase-3 expression was up-regulated,p-ERK1/2 expression was down-regulated,and Bcl-2/Bax ratio was decreased in group P (P＜0.05).Compared with group CO,the escape latency was significantly prolonged,the apoptosis rate was increased,the cleaved caspase-3 expression was up-regulated,p-ERK1/2 expression was down-regulated,and Bcl-2/Bax ratio was decreased in group PCO (P＜0.05).Conclusion The mechanism by which exogenous CO improves cognitive function is related to raising the phosphorylation of ERK1/2 in hippocampal neurons and inhibiting neuronal apoptosis in a rat model of hemorrhage shock and resuscitation.
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To develop a simple and easy promotion risk score to identify individuals with undiagnosed papillary thyroid carcinoma (PTC), and further to compare the diagnostic efficiency of PTC risk sore with fine needle aspiration cytodiagnosis (FNAC). in order to optimize the screening process of PTC. A sample of 1003 individuals aged 11-82 years underwent a surgical treatment of thyroid nodule participated in the study. The risk score was developed by stepwise backward multiple logistic regression. And using the receiver operating characteristic (ROC) curve to evaluate the diagnostic efficacy, the best diagnostic cut-off point and the risk stratification of malignant. Compare the sensitivity, specificity, accuracy, and area under curve ( AUC) of PTC risk score, FNAC and their combined diagnosis to judge their diagnostic efficiency. The risk score included age, TSH, nodule morphology and boundary by palpation, nodule characteristics, shape, boundary, calcification and blood flow signal by ultrasound. Its AUC= 0. 815, 6 point was the best cutoff point to differentiate benign from malignant thyroid nodules, and risk stratification of thyroid carcinoma were divided into four levels: very high risk group (score ≥ 9 points), high risk group ( score were 5-8 points), moderate risk group ( score were 3 ~ 4 points), low risk group ( score ≤ 2 points). The sensitivity, specificity, and accuracy of FNAC were 86. 3% , 90. 0% , and 87. 0% respectively, AUC=0. 891, while the sensitivity, specificity, and accuracy of PTC risk rating scale were 83. 8% , 70. 0% , and 81. 0%respectively, AUC = 0. 822. The sensitivity, specificity, and accuracy of their combined diagnosis were 97. 5% , 85. 0% , and 95. 0% , AUC=0. 965. This risk score can be used as a screening method before FNAC. If combined with FNAC, it may improve the diagnostic efficacy of PTC, and thereby possibly minimizing the unnecessary invasive examination and surgical treatment for patients with thyroid nodules and reducing personal costs.
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To develop a simple and easy promotion risk score to identify individuals with undiagnosed papillary thyroid carcinoma (PTC), and further to compare the diagnostic efficiency of PTC risk sore with fine needle aspiration cytodiagnosis (FNAC). in order to optimize the screening process of PTC. A sample of 1003 individuals aged 11-82 years underwent a surgical treatment of thyroid nodule participated in the study. The risk score was developed by stepwise backward multiple logistic regression. And using the receiver operating characteristic (ROC) curve to evaluate the diagnostic efficacy, the best diagnostic cut-off point and the risk stratification of malignant. Compare the sensitivity, specificity, accuracy, and area under curve ( AUC) of PTC risk score, FNAC and their combined diagnosis to judge their diagnostic efficiency. The risk score included age, TSH, nodule morphology and boundary by palpation, nodule characteristics, shape, boundary, calcification and blood flow signal by ultrasound. Its AUC= 0. 815, 6 point was the best cutoff point to differentiate benign from malignant thyroid nodules, and risk stratification of thyroid carcinoma were divided into four levels: very high risk group (score ≥ 9 points), high risk group ( score were 5-8 points), moderate risk group ( score were 3 ~ 4 points), low risk group ( score ≤ 2 points). The sensitivity, specificity, and accuracy of FNAC were 86. 3% , 90. 0% , and 87. 0% respectively, AUC=0. 891, while the sensitivity, specificity, and accuracy of PTC risk rating scale were 83. 8% , 70. 0% , and 81. 0%respectively, AUC = 0. 822. The sensitivity, specificity, and accuracy of their combined diagnosis were 97. 5% , 85. 0% , and 95. 0% , AUC=0. 965. This risk score can be used as a screening method before FNAC. If combined with FNAC, it may improve the diagnostic efficacy of PTC, and thereby possibly minimizing the unnecessary invasive examination and surgical treatment for patients with thyroid nodules and reducing personal costs.
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Objective To investigate the expression level and clinical significance of long non-coding RNA(LncRNA) growth arrest specific gene-antisense 1(GAS8-AS1) in papillary thyroid microcarcinoma(PTMC) patients. Methods We investigated the expression profile of GAS8-AS1 in tissue samples of patients with PTMC as well as nodular goiter(NG) by quantitative real-time polymerase chain reaction(RT-qPCR). Results GAS8-AS1 in cancer tissue was down-regulated in PTMC patients compared with adjacent thyroid tissue and NG samples(P<0.05). Lower level of GAS8-AS1 was also correlated with central cervical lymph node metastasis(CLNM, P<0.05). The area under the ROC curve for GAS8-AS1 was up to 0.717 3 in CLNM prediction(P<0.05). Conclusion GAS8-AS1 may act as a potential biomarker for PTC diagnosis and CLNM prediction.
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Objective To evaluate the role of mitochondrial ATP-seusitive potassium (mito-KATP) channels in sevoflurane postconditioning-induced inhibition of oxygen-glucose dcprivation and restoration (OGD/R)-induced pyroptosis in primary rat cardiomyocytes.Methods Cardiomyocytes of newborn Sprague-Dawley rats (＜48 h after birth) were cultured in vitro and seeded in 6-well dishes (2 cm in diameter)or in 96-well plates.The cells were divided into 6 groups (n =15 each) using a random number table:control group (group C),OGD/R group (group O),sevoflurane postconditioning group (group Sev),sevoflurane postconditioning plus 5-hydroxydecanoate (5-HD) group (group SH),5-HD group (group H) and OGD/R plus 5-HD group (group HO).The cardiomyocytes were subjected to oxygen-glucose deprivation for 4 h followed by restoration of oxygen-glucose supply for 24 h.After oxygen-glucose restoration,the cardiomyocytes in the culture media were exposed to 2％ sevoflurane for 1 h to perform sevoflurane postconditioning.At 1 h before oxygen-glucose deprivation,a specific mito-KATP channel blocker 5-HD 100 μmol/L was added to the culture media.Cardiomyocytes were cultured in normal culture atmosphere in group C.Cardiomyocytes were collected at 24 h of oxygen-glucose restoration.Cell pyroptosis was detected by double flow cytometry AlexaFour488 (caspase-1 FLICA staining) and TMR red (DNA staining) staining.The pyroptosis rate was calculated.The cell survival rate was measured by methyl thiazolyl tetrazolium assay.The content of reactive oxygen species (ROS) in mitochondria was determined by 2',7'-dichlorofluorescin diacetate assay.The mitochondrial membrane potential (MMP) was measured by using JC-I fluorescent probe.The expression of interleukin-1beta (IL-1β) was determined by Western blot.Results Compared with group C,the pyroptosis rate and ROS content were significantly increased,the cell survival rate and MMP were decreased,and the expression of IL-1β was up-regulated in group O (P＜0.05).Compared with group O,the pyroptosis rate and ROS content were significantly decreased,the cell survival rate and MMP were increased,and the expression of IL-1β was down-regulated in group Sev (P＜0.05).Compared with group Sev,the pyroptosis rate and ROS content were significantly increased,the cell survival rate and M MP were decreased,and the expression of IL-1β was up-regulated in group SH (P＜0.05).Compared with group SH,the pyroptosis rate and ROS content were significantly increased,the cell survival rate and MMP were decreased,and the expression of IL-1β was up-regulated in group H O (P＜0.05).Conclusion The mechanism by which sevoflurane postconditioning inhibits OGD/R-induced pyroptosis in primary rat cardiomyocytes is probably associated with increasing mito-KATP channel opening.
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Objective To establish a risk rating scale for central lymph node metastasis in papillary thyroid carcinoma and make risk stratification.Methods Data of 502 patients with PTC who were treated in Beijing Shijitan Hospital between January 2010 and June 2015 were retrospectively analyzed.The independent predictors for CLNM were found.Then a risk rating scale was established and stratification risk was made.The diagnostic value of the risk rating scale in predicting CLNM was evaluated.Data of 100 patients with PTC who were treated in Beijing Shijitan Hospital between July 2016 and June 2016 were used to validate the risk rating scale.Results A cutoff value of 5 points was found to be the best prediction for CLNM,with the sensitivity and specificity were 73.8％ and 70.2 ％.We definite score ≤ 4.5 as low risk for CLNM,as well as score from 5 to 7 as middle risk,score ≥ 7.5 as high risk.The other data of 100 patients was used to validate the risk rating scale.The sensitivity and specificity were 79.5％ and 78.7 ％ respectively.The positive predictive value and the negative predictive value were 70.5％ and 85.7％ respectively.Conclusions The risk rating scale provide a convenient,intuitive and quantized method to predict CLNM,which is helpful to select suitable surgical strategy and reduce operative complications.