Objective To investigate the organ protective role and the underlying mechanism of nafamostat mesylate(NM)in a renal ischemia-reperfusion injury(RIRI)model.Methods A total of 21 healthy male Sprague-Dawley(SD)rats were randomly assigned to 3 groups(n=7 in each group),including the sham operation group(Sham group),the RIRI group,and the NM intervention group(NM group).The RIRI and NM groups underwent ischemia-reperfusion injury(IRI)modeling.The NM group was given an intraperitoneal injection of NM at 0.75 mg/kg before modeling.Venous blood and renal tissue samples were then collected from the rats 24 hours after modeling.The levels of serum creatinine,cystatin C,and serum inflammatory factors were determined using the serum samples.Hematoxylin-eosin(HE)staining and TUNEL stainings were performed on the renal tissues to evaluate the damage of the renal tissues.The localization and expression of HMGB1 were analyzed by immunofluorescence and Western blotting,respectively.Single-cell RNA sequencing of the nuclei was performed to obtain the single-cell transcriptome of the kidneys from the rats in the RIRI and the NM groups and to acquire the RIRI cell profile.The cells were annotated according to the cell marker genes to explore the cell type composition in the disease model and the functional status of immune cells between the groups.Results 1)Compared with those of the Sham group,the levels of cystatin C,creatinine,and inflammatory factors in the RIRI and NM groups were significantly increased,and the expression levels in the NM group were lower than those in the RIRI group(P＜0.05).Compared with those of the RIRI group,the tubular injury score and apoptosis rate in the NM group were significantly decreased(P＜0.05),but those of both the NM and RIRI groups were higher than those of the sham group.Compared with that in the RIRI group,the expression of HMGB1 in the NM group was significantly decreased(P＜0.05),but the expression levels in both the RIRI and NM groups were higher than that in the sham group.Immunofluorescence showed that there was increased cytoplasmic expression of HMGB1 in both the NM and RIRI groups,with the increase being more prominent in the RIRI group.2)A total of 13 major cell populations were identified through the single-nucleus sequencing results.The proportion of tubular cells in the NM group was higher,with the HMGB1 gene being highly expressed in the damaged proximal convoluted tubular cells.The proportion of the polarized Macro3 cell subpopulation in the macrophages in the NM group was lower compared to that in the RIRI group.Conclusion NM may play a protective role in a rat model of RIRI,and its underlying mechanisms may be related to the regulation of the functional abnormalities of HMGB1-mediated macrophages.

Objective To explore the candidate genes and potential molecular mechanisms of anti-neutrophil cyto-plasmic antibodies(ANCA)-associated vasculitis by bioinformatics and experimental validation,and to provide a scientific theoretical basis for the treatment of potential inflammatory targets for ANCA-associated vasculitis.Meth-ods The GSE108109 chip data was retrieved from the Gene Expression Omnibus(GEO)database,and the differ-ential genes were processed,analyzed and screened using the R language related program package.Kyoto encyclo-pedia of genes and genomes(KEGG)and gene ontology(GO)enrichment analysis was carried out using DAVID online network cable,and the interaction network of the protein encoded by the selected genes of inflammatory syn-drome was constructed through STRING website.Further endogenous competitive RNA(ceRNA)regulatory net-work was predicted and constructed through miRWalk and DIANA-LncBase databases,and key genes were screened from the network to draw ROC curve.The renal biopsy samples of patients with ANCA-associated vasculi-tis confirmed by our hospital were collected as the experimental group,and the renal biopsy samples of IgA ne-phropathy and micro-adaptive nephropathy were collected as the control group.Immunohistochemical staining was performed on the collected renal biopsy samples,and the average optical density was calculated by semi-quantita-tive analysis of immunohistochemical staining to further verify the expression of the key genes screened by the bioin-formatics analysis.Pearson linear correlation analysis was performed between the average optical density results and the clinical inflammatory data of patients.Results 846 differential genes were screened,of which 444 genes were significantly up-regulated and 402 genes were significantly down-regulated.Through KEGG and GO analysis,im-portant differentially expressed genes related to inflammation regulation were obtained.Among them,CSF1R and TNFRSF1B,two differentially expressed genes never reported in ANCA-associated vasculitis,attracted our atten-tion.At the same time,we constructed multiple ceRNA regulatory axes including KCNQ1OT1-hsa-miR-125a-5p-TNFRSF1B.There were 15 samples of ANCA-associated vasculitis,6 samples of IgA nephropathy,and 3 samples of micropathological kidney.Immunohistochemical results of renal biopsy specimens showed that the expression of CSF1R and TNFRSF1B in ANCA-associated vasculitis kidney tissue was higher than that in the control group.Pearson correlation analysis of clinical data of patients in ANCA group showed that the expression of CSF1R was positively correlated with the content of neutrophil count(r=0.587),and the expression of TNFRSF1B was posi-tively correlated with the content of serum C-reactive protein(r=0.646).Conclusion Key genes related to in-flammatory regulation such as CSF1R and TNFRSF1B were investigated by bioinformatics methods,and a rigorous ceRNA regulatory network was constructed.The expression of CSF1R and TNFRSF1B in ANCA vasculitis was high-er than that in the control group through immunohistochemistry.The results provides a scientific theoretical basis for the molecular mechanism of inflammation,and laid a good foundation for new therapeutic targets of ANCA-related vasculitis for inflammation.

OBJECTIVE: To establish a prediction model of acute kidney injury (AKI) in moderate and severe burn patients, so as to provide basic research evidence for early identification of burn-related AKI. METHODS: Patients who were admitted to the department of plastic burn surgery of the Affiliated Hospital of Southwest Medical University from November 2018 to January 2021 were selected, and their clinical characteristics, laboratory examinations and other indicators were recorded. Multivariate Logistic regression analysis was used to screen out the risk factors of AKI related to moderate and severe burns, and R software was used to establish the nomogram of moderate and severe burn patients complicated with AKI. The Bootstrap method model was used for internal verification by repeating sample for 1 000 times. Consistency index and calibration curve were used to evaluate the accuracy of the model, and the receiver operator characteristic curve (ROC curve) and the area under the curve (AUC) were used to evaluate the prediction efficiency, decision curve analysis (DCA) was used to evaluate the clinical utility of the model. RESULTS: A total of 186 patients with moderate and severe burn were included, among which 54 patients suffered from AKI, and the incidence rate was 29.03%. Multivariate Logistic regression analysis showed that the total burn surface area [TBSA; odds ratio (OR) = 1.072, 95% confidence interval (95%CI) was 1.031-1.115, P = 0.001], estimated glomerular filtration rate (eGFR; OR = 0.960, 95%CI was 0.931-0.990, P = 0.010), neutrophil (NEU; OR = 1.190, 95%CI was 1.021-1.386, P = 0.026), neutrophil/lymphocyte ratio (NLR; OR = 0.867, 95%CI was 0.770-0.977, P = 0.019), D-dimer (OR = 4.603, 95%CI was 1.792-11.822, P = 0.002) were the risk factors for patients with moderate and severe burn complicated with AKI. Taking the above indexes as predictive factors, a nomogram prediction model was established, the ROC curve was plotted with AUC of 0.998 (95%CI was 0.988-1.000). Optimum threshold of ROC curve was -0.862, the sensitivity was 98.0% and the specificity was 98.2%, and the consistency index was 0.998 (95%CI was 0.988-1.000). The calibration curve showed that the prognostic nomogram model was accurate, DCA showed that most patients can benefit from this model. CONCLUSIONS The burned patients with higher TBSA, NEU, NLR, D-dimer and lower eGFR tend to suffer from AKI. The nomogram based on the above five risk factors has high accuracy and clinical value, which can be used as a predictive tool to evaluate the risk of AKI in moderate and severe burn patients.
Humans ; Prognosis ; Nomograms ; Retrospective Studies ; Burns/complications* ; Acute Kidney Injury/etiology* ; ROC Curve

OBJECTIVES: Cervicocerebral artery dissection (CAD) is one of the important causes for ischemic stroke in young and middle-aged people. CAD is dangerous and untimely diagnosis and treatment are likely to result in severe disability. Early diagnosis and timely intervention can greatly improve the prognosis of patients. This study was to investigate the imaging features of CAD on high-resolution magnetic resonance vessel wall imaging (HRMR-VWI) and to analyze the influential factors of vascular recanalization. METHODS: A total of 19 CAD patients with both baseline HRMR-VWI and follow-up data of vascular imaging in the period from April 2017 to December 2019 in Department of Radiology, Xiangya Hospital, Central South University were retrospectively analyzed. The diseased vessels were divided into a recovery group and a unrecovered group. After treatment, diseased vessels with no residual arterial dissection and no residual stenosis in the lumen were included in the recovery group. Diseased vessels with stenosis, occlusion or residual dissection were included in the unrecovered group. Diseased vessels were divided into a ischemic stroke group and a non-ischemic stroke group according to the presence or absence of ischemic stroke in the area supplied by the diseased vessels. Differences in clinical data and HRMR-VWI imaging findings were compared between the groups. RESULTS: A total of 26 vessels were involved, including 14 (53.8%) internal carotid artery extracranial segment, 8 (30.8%) vertebral artery extracranial segment, 3 (11.5%) vertebral artery intracranial segment, and 1 (3.9%) basilar artery. Ischemic stroke occurred in 16 diseased vascular supply areas. Intramural hematoma was all observed in the baseline HMR-VWI of the affected vessels. There were 18 vessels (69.2%) in the recovery group and 8 vessels (30.8%) in the unrecovered group. Compared with the vessels in the recovery group, the vessels in the unrecovered group were mostly found in the intracranial segment ( CONCLUSIONS Intramural hematoma is a common imaging manifestation of CAD and can be shown clearly and accurately on HRMR-VWI. Recanalization rate of CAD is high, and the recanalization of CAD in intracranial segment is slower than that of CAD in extracranial segment, which can prolong the review time.
Basilar Artery ; Dissection ; Humans ; Magnetic Resonance Angiography ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; Middle Aged ; Retrospective Studies ; Stroke

Objective To observe the differential expression of high mobility group box - 1 protein (HMGB1) in renal tissues of heat stroke mice models, and to explore its role in the pathogenesis of heat stroke associated acute kidney injury(HS-associated AKI). Methods According to random number table, 20 healthy male C57BL/6J mice were randomly divided into 2 groups, including normal control (n=10) and heat stroke group (n=10). The mice in heat stroke group were given with a 2-hour-exposure in biological simulation chamber (temperature 41℃, humidity 70％). Heat stroke was defined as anal temperature lasting more than 40 degrees Celsius. A 18F - deoxyglucose nuclide labeled vivo imaging was conducted with micro - positron emission tomography(PET)/computer tomography (CT). Serum creatinine was examined with blood example. In order to evaluate the pathological changes, HE stain was conducted with kidney tissue, and mitochondrial morphological changes in kidney tissue were observed by transmission electron microscopy. The expressions of HMGB1 and apoptosis inducing factor mitochondria associated 2 (Aifm2) were examined by immunohistochemical method, and the levels of HMGB1 and RAGE were examined by Western blotting. The cell apoptosis of renal tissue was detected by terminal deoxynucleotidyl transferase -mediated dUTP - biotin nick end labeling assay (TUNEL). The metabolomics of kidney tissue in mice were detected by liquid chromatography - mass spectrometry (LC - MS), and the pathway enrichment analysis was carried out by KEEG database. Results (1) The body temperature of the mice in heat shock group was significantly higher than that in normal control group 45 min after model establishment (P＜0.05). The level of serum creatinine in heat shock group was significantly higher than that in normal control group (P＜0.05), and the levels of 18F - deoxyglucose increased in skeletal muscle and visceral tissue of the mice in heat - shock group. (2) HE staining showed hemorrhage in collecting duct and tubular endothelial cell swelling, and mitochondrial swelling and deformation were observed by transmission electron microscopy in kidney tissue of the heat shock group. (3) Immunohistochemical method showed that the levels of Aifm2 and HMGB1 in heat shock group were higher (P＜0.05). (4) Western blotting showed that the levels of HMGB1 and RAGE in heat shock group were higher than those in normal control group (P＜0.05). (5) TUNEL showed that the number of cells with positive stain in kidney tissue of the heat shock group was higher than that in normal control group (P＜0.05). (6) Between normal control group and heat shock group, 136 differential metabolites were detected in kidney tissues. After analysis by KEGG database, pathway abnormalities such as unsaturated fatty acid metabolism disorder may be associated with HS - associated AKI, and many differential metabolites such as adrenic acid may be important regulatory points in the pathogenesis. Conclusion Acute kidney injury is a common complication of heat shock. It may be related to the dysfunction of renal mitochondria and activation of apoptotic pathway caused by systemic hypercatabolism, which may be related to the disorder of unsaturated fatty acid metabolism and activation of HMGB1. Some differential metabolites may be of high value in HS- associated AKI studies.

Objective To observe the expression of Notch1 signaling pathway in the aorta of chronic kidney disease (CKD) rats with vascular calcification and to explore the role of this signaling pathway activation in aortic calcification of CKD.Methods A total of 40 male SD rats were randomly divided into normal group (Nor group) and CKD with vascular calcification group (CKD+VC group).Rats in each group were sacrificed at 4 weeks,6 weeks and 8 weeks respectively after the success of modeling.Their 24-hour urine was reserved to test 24 hour urine protein (24 h Upro);blood sample was collected from abdominal aorta to test blood urea nitrogen (BUN),serum creatinine (Scr),Ca and P.The histopathology of renal was detected by HE staining.The aortic calcification was detected by alizarin red S staining.Immunohistochemistry (IHC) was used to test the protein expressions of alpha-smooth muscle actin (or-SMA),Runt-related transcription factor 2 (Runx2),Notchl,recombination signal-binding protein for immunoglobulin kappa J region (RBP-Jκ),Msh homeobox 2 (Msx2),Jagged1 and Notch1 intracellular domain (N1-ICD) in the aorta.Real time PCR was applied to detect the mRNA expressions of α-SMA,smooth muscle 22 alpha (SM22α),Runx2,alkaline phosphatase (ALP),Notch1,RBP-Jκ,Msx2 and Jagged1.Results Compared with those in Nor group,24 h Upro,BUN and Scr increased in the CKD+VC group at 4th,6th and 8th weeks (all P ＜ 0.05).Numerous continuous calcified nodules were detected in the vascular wall of the CKD+VC group,but none in Nor group.As compared with Nor group,the expression of α-SMA was low,while the expression of Runx2 was relatively high in the CKD+VC rats at each time point (all P ＜ 0.05).The expressions of Notchl,RBP-Jκ,Msx2,Jaggedl and NI-ICD in the Nor group were slightly appeared in the aortic wall,while in CKD+VC group these signal protein expressions increased relatively during the experimental period (all P ＜ 0.05).As compared with Nor group,the expressions of α-SMA and SM22α mRNA were low,yet the expressions of Runx2 and ALP mRNA were high in the CKD+VC rats at each time point (all P ＜ 0.05).The mRNA levels of Notch1,RBP-Jκ,Msx2 and Jagged1 in the CKD+VC group at each time point were significantly up-regulated as compared with the Nor group (all P ＜ 0.05).Conclusions There exist phenotypic changes in smooth muscle cells and activations of Notch1/RBP-Jκ/Msx2 signaling pathway in CKD rats with vascular calcification.It may be one of the important signal transduction pathways.

Objective To analyze the clinical features and risk factors of death in patients with acute mushroom poisonings. Methods The clinical data of 210 patients with acute mushroom poisoning admitted to the Affiliated Hospital of Southwest Medical University from July 2013 to December 2016 and received follow-up for at least 6 months were retrospectively analyzed. The data included gender, age, hospitalization time, toadstool features, incubation period, clinical performance, laboratory indicators, and prognosis. According to the prognosis, the patients were divided into survival group and non-survival group, the clinical characteristics and organ or system involvement of the two groups were analyzed, and the risk factors of death in patients with acute mushroom poisoning were explored by univariate and Logistic regression analysis. Results All 210 patients were enrolled in the final analysis, with 172 patients (81.9%) in survival group, and 38 (18.1%) in non-survival group. Patients with an incubation period of 6-24 hours had the highest mortality [15.2% (32/210)]. Most toadstools were in white, red or yellow, with an intake of 20-500 g. More than 85% of patients had gastrointestinal reactions, and liver damage was the most common [58.1% (122/210)] in all patients. The patients with heart and nervous system damage had higher mortality [61.4% (27/44) and 61.3% (19/31)], and the more organs or systems involved, the higher the mortality was. Univariate analysis showed that incubation period ≥ 6 hours, white blood cell (WBC) ≥12×109/L, alanine aminotransferase (ALT)≥200 U/L, aspartate aminotransferase (AST) ≥ 200 U/L, lactate dehydrogenase (LDH) ≥ 500 U/L, prothrombin time (PT) ≥ 20 s, activated partial thrombin time (APTT) ≥ 40 s, prothrombin activity (PTA) ≤ 60%, Na+≤ 135 mmol/L, MB isoenzyme of creatine kinase (CK-MB) ≥ 5 μg/L and myoglobin (Mb) ≥ 100 μg/L were the risk factors of death in patients with acute mushroom poisoning. Multiple factors Logistic regression analysis showed that APTT ≥ 40 s had the greatest lethal risk and could increase the risk of death by 5.35 times [odds ratio (OR) = 6.35, 95% confidence interval (95%CI) = 1.24-32.44], indicating that APTT was an independent risk factor of death in patients with acute mushroom poisoning. Conclusions The mortality of acute mushroom poisoning was high, and liver was the mainly involved organ. The incubation period, WBC, ALT, AST,LDH, PT, APTT, PTA, Na+, CK-MB and Mb could be early indicators to evaluate the prognosis in patients with acute mushroom poisoning, and patients with APTT ≥ 40 s had the greatest lethal risk.

Objective To explore the effects of renal artery calcification on the renal function in type 2 diabetic ne-phropathy rats .Methods Rats were randomly divided into control group ( CON group ) , diabetic nephropathy group ( DN group) and DN with vascular calcification group ( DN+VC group) .Rats of group DN and DN +VC were fed with high sugar and fat diet and injected with streptozotocin (STZ)into abdominal cavity to induce type 2 diabetes. After diabetic models were made , rats of group DN+VC were treated by vitamin D 3 plus nicotine .The rats were sacrificed at 8 , 12 and16 week respectively and the pathologic change to the renal artery were microscoped by von Kossa staining .The calcium content were detected by calcium assay kit and double immunofluorescence staining and real-time polymerase chain reaction ( RT-qPCR) were applied to detect the protein and gene expression levels of BMP2 in the renal artery.Measure the levels of blood urea nitrogen (BUN),serum creatinine (Scr),cystatin C (Cys C) and 24 hour urinary protein (24-h UA)respectively at the 8th,12th and 16th weeks.Histopathology of kidney was assessed by hematoxylin/eosin staining .Results The deposition of black granules , the calcium content and the protein and gene expression levels of BMP 2 in DN group were significantly higher than those in group CON and lower than DN+VC group at each time points(P<0.05).The BUN, Scr, Cys C and 24-h UA in group DN and group DN+VC were gradually increased in 8th,12th and 16th weeks, and were higher than those in group CON( P<0.05 ) .Compared with the DN group , only the level of Cys C at each time point and the level of 24-h UA in 16th week in DN+VC group were significantly higher ( P<0.05 ) .The pathological damages of the kidney in group DN showed a continual worsening trend and the pathological changes of the kidney in group DN +VC were more serious than group DN .Calcium content was positively correlated with the increased serum BUN , Scr, Cys C, 24-h UA and BMP2 mRNA ( all P<0.01 ) .Conclusions The occurrence and severity of renal artery calcification may participate in and promote the progression of DN .

Objective To analyze the coincidence of the patients with capillary electrophoresis hemoglobin A 2 increase with defi-nitely diagnosed beta thalassaemia and to investigate its application value in the diagnosis of beta thalassaemia.Methods Two hundreds and sixty outpatients and inpatients with hemoglobin A 2 increase in our hospital from May 2014 to May 2015 were per-formed the genetic testing.Results Among 260 patients with hemoglobin A2 increase ,beta thalassemia gene mutations were detec-ted in 257 cases ,the coincidence rate reached 98.85% ,and the common 17 beta thalassemia gene mutations were not detected in the other 3 cases ,follow-up further detection of rare beta thalassaemia gene and beta globin gene sequencing was performed ,1 case of SEA-HPFH βdeletion type was found ,1 case was Taiwaneseβdeletion type and 1 case was Codon 89-93(-AGT GAG CTG CAC TG) heterozygous mutation ,it was verified that 3 cases of hemoglobin A2 increase without detecting 17 kinds of common beta thalassemia gene mutations were still beta chain mutation occurrence or big fragment deletion.At the same time ,among 257 speci-mens of beta thalassemia gene mutations ,42 cases were compound alpha thalassaemia ,accounting for 16.34% of beta thalassaemia. Conclusion Capillary electrophoresis hemoglobin A2 increase can provide a fast and accurate basis for beta thalassemia diagnosis , but which can not rule out the possibility of compoound alpha thalassaemia ,when the patient's hemoglobin A2 is increased ,alpha and beta thalassaemia genetic diagnosis should be simultaneously carried out.

Objective Discussion the relationship of PALB2 and AR expression in triple negative breast cancer and its clinical pathological features. Methods The SP immunohistochemical staining was adopted to detect the expression of PALB2 and AR in 178 cases of TNBC, PALB2 divided into two groups according to the expression of different, and to analyze the clinical pathological features and prognostic impact of different AR expression status. Results 178 cases of TNBC, that PALB2 missing expression 47 cases (26.4%), AR expression of 60 cases (33.7%), between of them were negatively correlated (-1≤r<1, P<0.05), in PALB2 negative group, AR expression associated with family history, lymph node metastasis, clinical stage and recurrence and metastasis (P<0.05), and 5-year disease-free survival lower than AR negative expression, Log rank = 4.453, P = 0.035. Conclusion PALB2 negative expression while AR positive expression in TNBC have synergistic effect with disease progression, PALB 2 and AR combined detection may provide a new basis for the prognosis of TNBC interpretation, and recommending take further studies to confirm.