Comparative proteomics analysis of differential proteins in respond to doxorubicin resistance in myelogenous leukemia cell lines.

BACKGROUND: Chemoresistance remains a significant challenge in chronic myelogenous leukemia (CML) management, which is one of the most critical prognostic factors. Elucidation the molecular mechanisms underlying the resistance to chemoresistance may lead to better clinical outcomes. RESULTS: In order to identify potential protein targets involved in the drug-resistant phenotype of leukemia, especially the chronic myelogenous leukemia (CML), we used a high-resolution "ultra-zoom" 2DE-based proteomics approach to characterize global protein expression patterns in doxorubicin-resistant myelogenous leukemia cells compared with parental control cells. Ultra-high resolution of 2DE was achieved by using a series of slightly overlapping narrow-range IPG strips during isoelectric focusing (IEF) separation. A total number of 44 proteins with altered abundances were detected and identified by MALDI-TOF or LC-MS/MS. Among these proteins, enolase, aldolase, HSP70 and sorcin were up-regulated in doxorubicin-resistant myelogenous leukemia cell line, whereas HSP27 was down-regulated. Some of the results have been validated by Western blotting. Both enolase and aldolase were first reported to be involved in chemoresistance, suggesting that process of glycolysis in doxorubicin-resistant myelogenous leukemia cells was accelerated to some extent to provide more energy to survive chemical stress. Possible roles of most of the identified proteins in development of chemoresistance in myelogenous leukemia cells were fully discussed. The results presented here could provide clues to further study for elucidating the mechanisms underlying drug resistance in leukemia. CONCLUSIONS: As a whole, under the chemical stress, the doxorubicin-resistant myelogenous leukemia cells may employ various protective strategies to survive. These include: (i) pumping the cytotoxic drug out of the cells by P-glycoprotein, (ii) increased storage of fermentable fuel, (iii) sophisticated cellular protection by molecular chaperones, (iv) improved handling of intracellular calcium, (v) increased glucose utilization via increased rates of glycolysis. In the present study, proteomic analysis of leukemia cells and their drug resistant variants revealed multiple alterations in protein expression. Our results indicate that the development of drug resistance in doxorubicin-resistant myelogenous leukemia cells is a complex phenomenon undergoing several mechanisms.

The role and regulatory mechanism of RNA binding protein ZFP36in hypoxia/reoxygenation injury of cardiomyocytes / 西安交通大学学报(医学版)

【Objective】 To explore the role of ZFP36 in cardiomyocyte injury and autophagy induced by hypoxia/reoxygenation (H/R) so as to clarify its molecular regulatory mechanism. 【Methods】 H9C2 rat cardiomyocytes were infected with ZFP36 overexpressing lentivirus (OE-ZFP36) or its negative control lentivirus (OE-ZFP36 NC) to construct stable cell lines, respectively. Transfection of ATG4D overexpression plasmid (OE-ATG4D) improved the expression of ATG4D. Hypoxia/reoxygenation (H/R) induced myocardial cell injury. H9C2 cells were mainly divided into control group, H/R group, OE-ZFP36 NC+H/R group, OE-ZFP36+H/R group, OE-ATG4D NC+H/R group, OE-ATG4D+H/R group, OE-ZFP36+OE-ATG4D NC+H/R group, and OE-ZFP36+OE-ATG4D+H/R group. The protein expressions of ATG4D, Beclin1, LC3 and ZFP36 in H9C2 cells were detected by Western blotting. The mRNA levels of ZFP36 and ATG4D in H9C2 cells were detected by Real-time fluorescence quantitative PCR (qPCR). The viability of H9C2 cells was detected by CCK-8 assay. The levels of interleukin (IL-6) and tumor necrosis factor (TNF-α) in H9C2 cells were detected by enzyme-linked immunosorbent assay (ELISA). Reactive oxygen species (ROS) in H9C2 cells were detected by DCFH-DA method. SOD detection kit was used to detect the SOD level in H9C2 cells. The apoptosis of H9C2 cells was detected by flow cytometry. LC3 autophagosomes in H9C2 cells were detected by cellular immunofluorescence. Dual-luciferase reporter gene assay was used to detect the binding of ZFP36 and ATG4D mRNA in H9C2 cells. 【Results】 Compared with control group, H/R group showed decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels and decreased SOD levels, increased cell apoptosis. Up-regulated ATG4D and Beclin1 protein expression, increased LC3Ⅱ/LC3Ⅰ ratio, as well as upregulated ZFP36 expression were found in H/R group (all P<0.05). Compared with OE-ZFP36 NC+H/R group, elevated cell viability, decreased IL-6 and TNF-α levels, decreased ROS levels and increased SOD levels, reduced cell apoptosis (P<0.05), and downregulated ATG4D and Beclin1 protein expression, decreased LC3Ⅱ/LC3Ⅰ ratio were shown in OE-ZFP36+H/R group (all P<0.05). Compared with infection with OE-ZFP36 NC lentivirus, infection with OE-ZFP36 lentivirus decreased the luciferase activity of ATG4D 3′-UTR reporter gene, decreased the stability of ATG4D mRNA, and downregulated the H/R-induced ATG4D mRNA expression (all P<0.05). Compared with OE-ATG4D NC+H/R group, OE-ATG4D+H/R group had upregulated ATG4D mRNA and protein expression, decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels, decreased SOD levels and elevated cell apoptosis (all P<0.05). Compared with OE-ZFP36+OE-ATG4D NC+H/R group, OE-ZFP36+OE-ATG4D+H/R group had decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels, decreased SOD levels and elevated cell apoptosis (all P<0.05). 【Conclusion】 The expression of ZFP36 is upregulated in H/R-induced cardiomyocyte injury. The overexpression of ZFP36 inhibits H/R-induced cardiomyocyte injury and autophagy by regulating ATG4D, thus resisting cardiomyocyte H/R injury. It proves that ZFP36 is an important regulatory molecule against MI/RI.

Probing bundle-wise abnormalities in patients infected with human immunodeficiency virus using fixel-based analysis: new insights into neurocognitive impairments / 中华医学杂志(英文版)

BACKGROUND@#Changes in white matter (WM) underlie the neurocognitive damages induced by a human immunodeficiency virus (HIV) infection. This study aimed to examine using a bundle-associated fixel-based analysis (FBA) pipeline for investigating the microstructural and macrostructural alterations in the WM of the brain of HIV patients.@*METHODS@#This study collected 93 HIV infected patients and 45 age/education/handedness matched healthy controls (HCs) at the Beijing Youan Hospital between January 1, 2016 and December 30, 2016.All HIV patients underwent neurocognitive evaluation and laboratory testing followed by magnetic resonance imaging (MRI) scanning. In order to detect the bundle-wise WM abnormalities accurately, a specific WM bundle template with 56 tracts of interest was firstly generated by an automated fiber clustering method using a subset of subjects. Fixel-based analysis was used to investigate bundle-wise differences between HIV patients and HCs in three perspectives: fiber density (FD), fiber cross-section (FC), and fiber density and cross-section (FDC). The between-group differences were detected by a two-sample t -test with the false discovery rate (FDR) correction ( P <0.05). Furthermore, the covarying relationship in FD, FC and FDC between any pair of bundles was also accessed by the constructed covariance networks, which was subsequently compared between HIV and HCs via permutation t -tests. The correlations between abnormal WM metrics and the cognitive functions of HIV patients were explored via partial correlation analysis after controlling age and gender.@*RESULTS@#Among FD, FC and FDC, FD was the only metric that showed significant bundle-wise alterations in HIV patients compared to HCs. Increased FD values were observed in the bilateral fronto pontine tract, corona radiata frontal, left arcuate fasciculus, left corona radiata parietal, left superior longitudinal fasciculus III, and right superficial frontal parietal (SFP) (all FDR P <0.05). In bundle-wise covariance network, HIV patients displayed decreased FD and increased FC covarying patterns in comparison to HC ( P <0.05) , especially between associated pathways. Finally, the FCs of several tracts exhibited a significant correlation with language and attention-related functions.@*CONCLUSIONS@#Our study demonstrated the utility of FBA on detecting the WM alterations related to HIV infection. The bundle-wise FBA method provides a new perspective for investigating HIV-induced microstructural and macrostructural WM-related changes, which may help to understand cognitive dysfunction in HIV patients thoroughly.

Diagnosis of primary clear cell carcinoma of the liver based on Faster region-based convolutional neural network / 中华医学杂志(英文版)

BACKGROUND@#Distinguishing between primary clear cell carcinoma of the liver (PCCCL) and common hepatocellular carcinoma (CHCC) through traditional inspection methods before the operation is difficult. This study aimed to establish a Faster region-based convolutional neural network (RCNN) model for the accurate differential diagnosis of PCCCL and CHCC.@*METHODS@#In this study, we collected the data of 62 patients with PCCCL and 1079 patients with CHCC in Beijing YouAn Hospital from June 2012 to May 2020. A total of 109 patients with CHCC and 42 patients with PCCCL were randomly divided into the training validation set and the test set in a ratio of 4:1.The Faster RCNN was used for deep learning of patients' data in the training validation set, and established a convolutional neural network model to distinguish PCCCL and CHCC. The accuracy, average precision, and the recall of the model for diagnosing PCCCL and CHCC were used to evaluate the detection performance of the Faster RCNN algorithm.@*RESULTS@#A total of 4392 images of 121 patients (1032 images of 33 patients with PCCCL and 3360 images of 88 patients with CHCC) were uesd in test set for deep learning and establishing the model, and 1072 images of 30 patients (320 images of nine patients with PCCCL and 752 images of 21 patients with CHCC) were used to test the model. The accuracy of the model for accurately diagnosing PCCCL and CHCC was 0.962 (95% confidence interval [CI]: 0.931-0.992). The average precision of the model for diagnosing PCCCL was 0.908 (95% CI: 0.823-0.993) and that for diagnosing CHCC was 0.907 (95% CI: 0.823-0.993). The recall of the model for diagnosing PCCCL was 0.951 (95% CI: 0.916-0.985) and that for diagnosing CHCC was 0.960 (95% CI: 0.854-0.962). The time to make a diagnosis using the model took an average of 4 s for each patient.@*CONCLUSION@#The Faster RCNN model can accurately distinguish PCCCL and CHCC. This model could be important for clinicians to make appropriate treatment plans for patients with PCCCL or CHCC.

Development of a CLDN18.2-targeting immuno-PET probe for non-invasive imaging in gastrointestinal tumors / 药物分析学报

Claudin18.2(CLDN18.2)is a tight junction protein that is overexpressed in a variety of solid tumors such as gastrointestinal cancer and oesophageal cancer.It has been identified as a promising target and a potential biomarker to diagnose tumor,evaluate efficacy,and determine patient prognosis.TST001 is a recombinant humanized CLDN18.2 antibody that selectively binds to the extracellular loop of human Claudin18.2.In this study,we constructed a solid target radionuclide zirconium-89(89Zr)labled-TST001 to detect the expression of in the human stomach cancer BGC823CLDN18.2 cell lines.The[89Zr]Zr-des-ferrioxamine(DFO)-TST001 showed high radiochemical purity(RCP,＞99％)and specific activity(24.15±1.34 GBq/μmol),and was stable in 5％human serum albumin,and phosphate buffer saline(＞85％RCP at 96 h).The EC50 values of TST001 and DFO-TST001 were as high as 0.413±0.055 and 0.361±0.058 nM(P＞0.05),respectively.The radiotracer had a significantly higher average standard uptake values in CLDN18.2-positive tumors than in CLDN18.2-negative tumors(1.11±0.02 vs.0.49±0.03,P=0.0016)2 days post injection(p.i.).BGC823CLDN18.2 mice models showed high tumor/muscle ratios 96 h p.i.with[89Zr]Zr-DFO-TST001 was much higher than those of the other imaging groups.Immunohistochemistry results showed that BGC823CLDN18.2 tumors were highly positive(+++)for CLDN18.2,while those in the BGC823 group did not express CLDN18.2(-).The results of ex vivo biodistribution studies showed that there was a higher distribution in the BGC823CLDN18.2 tumor bearing mice(2.05±0.16％ID/g)than BGC823 mice(0.69±0.02％ID/g)and blocking group(0.72±0.02％ID/g).A dosimetry estimation study showed that the effective dose of[89Zr]Zr-DFO-TST001 was 0.0705 mSv/MBq,which is within the range of acceptable doses for nuclear medicine research.Taken together,these re-sults suggest that Good Manufacturing Practices produced by this immuno-positron emission tomog-raphy probe can detect CLDN18.2-overexpressing tumors.

Association between the Wnt signaling pathway and hepatic fibrosis / 临床肝胆病杂志

Hepatic fibrosis (HF) is a self-healing pathological process after all kinds of chronic liver injuries and can cause diseases such as liver cirrhosis and liver cancer. The Wnt signaling pathway is highly conserved in species evolution and widely exists in invertebrates and vertebrates, and many studies have confirmed that the Wnt signaling pathway is closely associated with the development and progression of HF. This article reviews the mechanisms of the classical and non-classical Wnt signaling pathways in regulating hepatic stellate cells, hepatic macrophages, and hepatic progenitor cells, so as to provide new ideas for subsequent studies on the mechanism of the Wnt signaling pathway in regulating HF and further exploration of therapeutic targets that can reverse HF.

A Recombinant HAV Expressing a Neutralization Epitope of HEV Induces Immune Response against HAV and HEV in Mice.

Hepatitis A virus (HAV) and hepatitis E virus (HEV) are causative agents of acute viral hepatitis transmitted via the fecal-oral route. Both viruses place a heavy burden on the public health and economy of developing countries. To test the possibility that HAV could be used as an expression vector for the development of a combination vaccine against hepatitis A and E infections, recombinant HAV-HEp148 was created as a vector to express an HEV neutralization epitope (HEp148) located at aa 459-606 of the HEV capsid protein. The recombinant virus expressed the HEp148 protein in a partially dimerized state in HAV-susceptible cells. Immunization with the HAV-HEp148 virus induced a strong HAV- and HEV-specific immune response in mice. Thus, the present study demonstrates a novel approach to the development of a combined hepatitis A and E vaccine.

Effect of different sufentanil doses on hemodynamics and interleukin-6 in children with ventricular septal defect repair / 中国医师进修杂志

Objective:To study the effect of different sufentanil doses on hemodynamics and interleukin (IL)-6 in children with ventricular septal defect repair (VSDR).Methods:A total of 96 children who underwent VSDR surgery in Three Gorges Hospital Affiliated to Chongqing University from January 2016 to June 2019 were selected. Children with VSDR were enrolled and divided into A group (0.8 μg/kg), B group (1.0 μg/kg) and C group (1.1 μg/kg) according their sufentanil doses. The heart rate (HR), contraction pressure (SBP), the brain electric double frequency index (BIS) and IL-6 in different time were studied and compared. The adverse cardiovascular events and other indicators were studied and compared.Results:At t 2 to t 6, there were significant difference among the three groups in HR, A group: (104.62 ± 10.58), (128.73 ± 13.29), (127.59 ± 13.53), (125.62 ± 12.60) and (118.49 ± 11.62) times/min, B group: (100.27 ± 10.11), (119.33 ± 12.62), (116.57 ± 11.40), (113.57 ± 11.94) and (113.37 ± 11.46) times/min, C group: (87.92 ± 8.87), (98.62 ± 9.69), (94.42 ± 9.38), (88.72 ± 8.62) and (89.36 ± 9.17) times/min; SBP, A group: (79.54 ± 7.59), (102.75 ± 10.62), (90.62 ± 9.52), (85.19 ± 8.62) and (83.98 ± 8.62) mmHg (1 mmHg = 0.133 kPa), B group: (76.74 ± 7.28), (90.45 ± 9.57), (87.38 ± 8.51), (84.72 ± 8.50) and (83.77 ± 8.58) mmHg, C group: (70.62 ± 7.27), (75.62 ± 7.83), (72.69 ± 7.80), (71.28 ± 7.43) and (71.39 ±7.16) mmHg, P<0.05. At t 2 to t 4, there were significant differences among the three groups in BIS, A group: 48.64 ± 5.03, 53.58 ± 5.71 and 59.61 ± 5.87, B group: 48.79 ± 5.12, 50.23 ± 5.24 and 57.75 ± 5.66, C group: 43.59 ± 4.62, 50.31 ± 5.34, 55.26 ± 5.53, P<0.05. At T 2 to T 6, there was a significant difference among the three groups in IL-6, A group: (0.41 ± 0.06), (0.49 ± 0.11), (0.53 ± 0.13), (0.82 ± 0.17) and (0.61 ± 0.15) μg/L, B group: (0.38 ± 0.05), (0.42 ± 0.08), (0.46 ± 0.09), (0.75 ± 0.14) and (0.53 ± 0.12) μg/L, C group: (0.35 ± 0.05), (0.40 ± 0.06), (0.43 ± 0.07), (0.72 ± 0.12) and (0.51 ± 0.11) μg/L, P<0.05. Compared with that in A group and C group, HR, SBP and BIS in B group had relatively low volatility. At T 2 to T 6, IL-6 level in A group was significantly higher than that in B group and C group, and there was a statistically significant difference ( P<0.05). The incidence of cardiovascular adverse events in A group and C group was significantly higher than that in B group, and there was statistical difference ( P<0.05). Conclusions:1.0 μg/kg sufentanil can have less effect on hemodynamics and IL-6 in children with VSDR with less adverse cardiovascular events and other indicators.

Dosimetry verfication between pre and post operation of 3D-printing template assisted by CT-guided 125I seed implantation in the treatment of bone metastases / 中华放射医学与防护杂志

Objective:To compare and analyze the differences of common dose indicators before and after operation of 3D-printing template assisted by CT-guided 125I seed implantation in the treatment of bone metastases to guide clinical application. Methods:A retrospective analysis of 12 lesions in the 10 patients (9 males and 1 female, median age 65 years), who underwent seed implantation surgery for bone metastases in the Tianjin Third Central Hospital from June 2019 to January 2021, was conducted. All the lesions were adopted for 3D-printing template to guide seed implantation and the prescribed dose was 120-140 Gy. The differences of common dose indicators between preoperative treatment plan and postoperative verification plan were compared, including D90 (dose received by 90% of the target volume), D100 (dose received by 100% of the target volume), V90 (the volume percent for tumor target volume receiving 90% of the prescribed dose), V100 (the volume percent for tumor target volume receiving 100% of the prescribed dose), V150 (the volume percent for tumor target volume receiving 150 % of the prescribed dose), as well as the quantity of seeds planned and actually used. The paired t-test was performed to compare and analyze those parameters. Results:There was no statistically significant difference in pre- and postoperative D90, D100, V90, V100, V150 ( P > 0.05). The actual quantity of seeds used after operation was more than that of the preoperative planned quantity and the difference was statistically significant ( t=-2.930, P < 0.05). Conclusions:The clinical use of 3D-printing template assisted by CT-guided 125I seed implantation for bone metastasis should be promoted since the dose is accurate and the requirements of preoperative plan can be achieved.

Research on the role of miR-125b-1-3p in rotavirus replication / 中华微生物学和免疫学杂志

Objective:To investigate the role and regulatory mechanism of miR-125b-1-3p in rotavirus replication.Methods:MA104 cells were infected with rotavirus after upregulation and down-regulation of miR-125b-1-3p, respectively. The expression of miR-125b-1-3p and the copy number of rotavirus were analyzed by RT-PCR. The effect of miR-125b-1-3p on the protein expression of rotavirus was analyzed by immunofluorescence. The expression of related proteins involved in the regulation of miR-125b-1-3p was analyzed by Western blot analysis.Results:After rotavirus infection, the expression level of miR-125b-1-3p was significantly up-regulated, the copy number of VP7 and NSP3 gene of rotavirus decreased after up-regulation of miR-125b-1-3p, and the copy number of VP7 and NSP3 gene of rotavirus was significantly increased after down-regulation of miR-125b-1-3p.The fluorescence number of rotavirus protein decreased after upregulation of miR-125b-1-3p expression level, and increased after down-regulation of miR-125b-1-3p expression level. The activity of PI3K/Akt pathway was inhibited 16 h after rotavirus infection, and the up-regulation of miR-125b-1-3p could inhibit the activation of PI3K/Akt pathway.Conclusions:MiR-125b-1-3p inhibits rotavirus replication by regulating the PI3K/Akt pathway. These results provide an experimental basis for exploring the specific regulatory mechanism between miR-125b-1-3p and PI3K/Akt pathway, and provide a target for anti-infection therapy of rotavirus.

Prognostic nomogram incorporating radiological features for predicting overall survival in patients with AIDS-related non-Hodgkin lymphoma / 中华医学杂志(英文版)

BACKGROUND@#Acquired immune deficiency syndrome (AIDS)-related non-Hodgkin lymphoma (AR-NHL) is a high-risk factor for morbidity and mortality in patients with AIDS. This study aimed to determine the prognostic factors associated with overall survival (OS) and to develop a prognostic nomogram incorporating computed tomography imaging features in patients with acquired immune deficiency syndrome-related non-Hodgkin lymphoma (AR-NHL).@*METHODS@#A total of 121 AR-NHL patients between July 2012 and November 2019 were retrospectively reviewed. Clinical and radiological independent predictors of OS were confirmed using multivariable Cox analysis. A prognostic nomogram was constructed based on the above clinical and radiological factors and then provided optimum accuracy in predicting OS. The predictive accuracy of the nomogram was determined by Harrell C-statistic. Kaplan-Meier survival analysis was used to determine median OS. The prognostic value of adjuvant therapy was evaluated in different subgroups.@*RESULTS@#In the multivariate Cox regression analysis, involvement of mediastinal or hilar lymph nodes, liver, necrosis in the lesions, the treatment with chemotherapy, and the CD4 ≤100 cells/μL were independent risk factors for poor OS (all P < 0.050). The predictive nomogram based on Cox regression has good discrimination (Harrell C-index = 0.716) and good calibration (Hosmer-Lemeshow test, P = 0.620) in high- and low-risk groups. Only patients in the high-risk group who received adjuvant chemotherapy had a significantly better survival outcome.@*CONCLUSION@#A survival-predicting nomogram was developed in this study, which was effective in assessing the survival outcomes of patients with AR-NHL. Notably, decision-making of chemotherapy regimens and more frequent follow-up should be considered in the high-risk group determined by this model.

Strategy for the management of patients after varicocelectomy / 中华泌尿外科杂志

With the improvement of surgical methods, more and more patients with varicocele accepted operations. The management of the patients after varicocelectomy needs to be strengthened. In the present review article, evaluation for curative outcomes and follow-up therapeutic selections were introduced systematically, including watchful waiting, non-operative methods, and reoperation.

Effect of leptin on pyroptosis in hippocampal cells during brain injury in a rat model of orthotopic liver transplantation / 中华麻醉学杂志

Objective:To evaluate the effect of leptin on pyroptosis in hippocampal cells during brain injury in a rat model of orthotopic liver transplantation (OLT).Methods:Fifty-four clean-grade male Sprague-Dawley rats, aged 12 weeks, weighing 200-250 g, were divided into 3 groups( n=18 each)by a random number table method: sham operation group (S group), OLT-induced brain injury group (Liver group) and leptin group (Lep group). The model of OLT was established according to the " two-cuff" technique in anesthetized rats in Liver and Lep groups.The perihepatic ligament was only isolated and freed in S group.Leptin 1 mg/kg was intraperitoneally injected immediately after portal vein occlusion in Lep group.The equal volume of normal saline was intraperitoneally injected at the same time point in S group and Liver group.Twelve rats in each group were selected on day 3 after surgery and sacrificed, and the hippocampal tissues were obtained for examination of the pathological changes (with a light microscope) and for determination of the expression of NOD-like receptor protein 3 (NLRP3), caspase-1, interleukin-1beta (IL-1β), IL-18 and gasdermin D (GSDMD) mRNA (by real-time polymerase chain reaction) and expression of NLRP3, caspase-1, IL-1β, IL-18 and GSDMD (by Western blot). Morris water maze test was performed in the remaining 6 rats in each group on day 30 after surgery. Results:Compared with S group, the count of viable neurons in hippocampi was significantly decreased, the expression of NLRP3, caspase-1, IL-1β, IL-18 and GSDMD protein and mRNA was up-regulated, the time of staying at the platform quadrant was shortened, and the escape latency was prolonged in Liver and Lep groups ( P<0.05). Compared with Liver group, the count of viable neurons in hippocampi was significantly increased, the expression of NLRP3, caspase-1, IL-1β, IL-18 and GSDMD protein and mRNA was down-regulated, the time of staying at the platform quadrant was prolonged, and the escape latency was shortened in Lep group ( P<0.05). The pathological changes of hippocampal tissues were significantly attenuated in Liver group when compared with Lep group. Conclusion:The mechanism by which leptin reduces brain injury is related to inhibiting pyroptosis in hippocampal cells in a rat model of OLT.

Progress on adenovirus-vectored universal influenza vaccines.

Influenza virus (IFV) infection causes serious health problems and heavy financial burdens each year worldwide. The classical inactivated influenza virus vaccine (IIVV) and live attenuated influenza vaccine (LAIV) must be updated regularly to match the new strains that evolve due to antigenic drift and antigenic shift. However, with the discovery of broadly neutralizing antibodies that recognize conserved antigens, and the CD8(+) T cell responses targeting viral internal proteins nucleoprotein (NP), matrix protein 1 (M1) and polymerase basic 1 (PB1), it is possible to develop a universal influenza vaccine based on the conserved hemagglutinin (HA) stem, NP, and matrix proteins. Recombinant adenovirus (rAd) is an ideal influenza vaccine vector because it has an ideal stability and safety profile, induces balanced humoral and cell-mediated immune responses due to activation of innate immunity, provides 'self-adjuvanting' activity, can mimic natural IFV infection, and confers seamless protection against mucosal pathogens. Moreover, this vector can be developed as a low-cost, rapid-response vaccine that can be quickly manufactured. Therefore, an adenovirus vector encoding conserved influenza antigens holds promise in the development of a universal influenza vaccine. This review will summarize the progress in adenovirus-vectored universal flu vaccines and discuss future novel approaches.

Quantitative evaluation of magnetic resonance T2 mapping in Brucella spondylitis / 中华地方病学杂志

Objective:To analyze the characteristics and quantitative evaluation of Brucella spondylitis patients by magnetic resonance T2 mapping. Methods:A prospective clinical study was conducted to analyze the MRI data of 23 patients with brucellosis spondylitis diagnosed in the First Affiliated Hospital of Xinjiang Medical University from January 2016 to September 2018, and 25 healthy volunteers were selected as the control group. MRI was used to examine the vertebral bodies of the subjects, and T2 mapping map was automatically generated. Regions of interest (ROI) were selected on the T2 mapping map to generate T2 mapping values automatically. The T2 mapping values of diseased vertebrae, adjacent unaffected vertebrae, paravertebral abscess and healthy volunteers were analyzed.Results:Among 48 MRI examinees, 23 cases were Brucella spondylitis, including 17 males and 6 females, aged (38.5 ± 13.4) years; 25 healthy volunteers, including 15 males and 10 females, aged (35.1 ± 12.7) years. In 23 patients with Brucella spondylitis, 5 thoracic vertebrae, 40 lumbar vertebrae and 7 sacral vertebrae were involved, with a total of 52 vertebral bodies. Among them, 11 (21.2%) vertebral bodies showed bone marrow edema on MRI, 41 (78.8%) vertebral bodies showed bone marrow edema and vertebral bone destruction. T2 mapping values of the diseased vertebrae, adjacent unaffected vertebrae and paravertebral abscess in Brucella spondylitis patients and normal vertebrae in healthy volunteers were (115.62 ± 11.37), (75.21 ± 5.57), (240.26 ± 30.67) and (77.29 ± 4.19) ms, respectively. There were significant differences between the diseased vertebrae in Brucella spondylitis and adjacent unaffected vertebrae in Brucella spondylitis, and there were significant differences between the diseased vertebrae in Brucella spondylitis and normal vertebrae in healthy volunteers ( t = 26.78, 19.42, P < 0.05). Conclusion:Magnetic resonance T2 mapping can be used to evaluate the pathological tissues in Brucella spondylitis patients, and it has certain guiding significance for the quantitative description and qualitative diagnosis.

Laboratory confirmation of an outbreak of respiratory infectious disease caused by human adenovirus 3 in Beijing / 中华微生物学和免疫学杂志

Objective:To confirm the possible pathogen causing an outbreak of respiratory infectious disease in Beijing.Methods:Oropharyngeal swabs were collected from 14 cases with fever and detected by RT-PCR for respiratory viruses and bacteria. For specimens positive for adenoviruses, Fiber, Hexon and Penton gene fragments were amplified with specific primers and sequenced. BLAST and phylogenetic tree were used for sequence analysis.Results:All of the 14 specimens were adenovirus-positive. BLAST analysis of the sequences of Fiber, hexon and Penton genes showed that the 14 cases were all caused by adenovirus 3. The phylogenic tree analysis indicated that this adenovirus was closely related to an adenovirus of 3a51 genotype (GenBank No: KF268123) isolated in the USA in 2007.Conclusions:Human adenovirus genotype 3a51 caused this outbreak of respiratory infectious disease in Beijing.

Effect of Danhong injection on expression of nephrin and podocin in podocyte of renal tissue in rats with membranous nephropathy / 国际中医中药杂志

Objective To observe the effect of Danhong injection on the expression of nephrin podocin protein and gene in the kidney of membranous nephropathy rats.Methods A total of 34 male clean level SD rats were divided at random into the normal control group (8 rats),model group (26 rates).The model group were injected the C-BSA into caudal vein to establish the model of the membranous nephropathy.After four week of succession of the models,two model rats were taken to confirm the success of the model.Another 24 models divided at random into model group,Losartan potassium treatment group and Danhong injection treatment group,8 in each group.In the losartan group,the losartan 40 mg/kg was administered,and the Danhong group was injected with 4 ml/kg of Danhong injection in the tail vein for 4 weeks.The rat urine was collected after the last administration,and 24 h urine protein was determined by spectrophotometer.The serum total protein,albumin,TC and TG levels were determined by automatic biochemical analyzer.The renal pathological changes were observed by HE staining.The expressions of nephrin,podocin and WT1 labeled podocytes were detected by immunohistochemical staining.The expressions of nephrin and podocin in renal tissue were detected by real-time quantitative PCR and Western blot,and the results were correlated.Results Compared with the model group,the content of serum albumin increased in the Danhong group (P＜0.01),24 h urine protein quantitation,TC,TG significantly decreased (P＜0.01).The expression of nephrin protein (1.12 ± 0.03 vs.0.57 ± 0.09),podocin protein (0.99 ± 0.04 vs.0.53 ± 0.08),nephrin mRNA (0.49 ± 0.08 vs.0.14 ± 0.02),podocin mRNA (0.50 ± 0.11 vs.1.18 ± 0.05) in Danhong group significantly increased (P＜0.01),and the number of podocytes labeled with WT1 (14.88％ ± 1.73％ vs.9.63％ ± 1.69％) significantly increased (P＜0.05).Correlation analysis showed that 24 h urine protein quantitation and expression of nephrin protein (r=-0.908),podocin protein (r=-0.916),nephrin mRNA (r=-0.930),podocin mRNA (r=-0.923).There was a negative correlation between serum albumin (r=-0.934) and podocyte count (r=-0.778) (P＜0.01),and positive correlation with TC (r=0.814) and TG (r=0.870) (P＜0.01).Conclusions The Danhong injection may reduce the urine protein of membranous nephropathy rats by up-regulating the expression of nephrin,podocin mRNA and protein in renal cells of membranous nephropathy.

Effects of leptin on brain injury and long-term cognitive function in rats undergoing orthotopic liver transplantation / 中华麻醉学杂志

Objective To evaluate the effects of leptin on brain injury and long-term cognitive function in rats undergoing orthotopic liver transplantation. Methods Ninety clean-grade male Sprague-Dawley rats, aged 3 months, weighing 200-250 g, were divided into 3 groups by a random number table method: sham operation group ( S group) , liver ischemia-reperfusion ( I∕R) group ( I∕R group) and lep-tin group ( L group) , with 18 rats in each group. Orthotopic liver transplantation was performed to estab-lish the model of liver I∕R injury in I∕R and L groups. Leptin 1 mg∕kg was intraperitoneally injected at the onset of ischemia in L group, and the equal volume of normal saline was given instead of leptin in S and I∕R groups. Twelve rats in each group were sacrificed at 3 days after operation, and brains were removed for ex-amination of the pathological changes in hippocampal CA1 region ( with a light microscope) and for determi-nation of apoptosis in hippocampal neurons ( by TUNEL assay) and expression of aquaporin 4 ( AQP4) and protein kinase C ( PKC) in the hippocampus ( by Western blot) . The apoptosis rate was calculated. The remaining 6 rats in each group underwent a Morris water maze test at 30 days after surgery to evaluate long-term cognitive function. Results Compared with S group, the apoptosis rate of hippocampal neurons was significantly increased, the expression of AQP4 and PKC was up-regulated, the escape latency was pro-longed, and the time of staying at the platform quadrant was shortened in I∕R and L groups (P<0. 05). Compared with I∕R group, the apoptosis rate of hippocampal neurons was significantly decreased, the ex-pression of AQP4 and PKC was down-regulated, the escape latency was shortened, and the time of staying at the platform quadrant was prolonged in L group (P<0. 05). Conclusion Leptin can reduce the brain damage in rats undergoing orthotopic liver transplantation, the mechanism may be related to down-regulating the expression of PKC and AQP4, and leptin can also improve long-term cognitive function after orthotopic liver transplantation in rats.

Effects of dexmedetomidine on brain injury and long -term cognitive dysfunction in rats after orthotopic liver transplantation ischemia/reperfusion / 中华神经医学杂志

Objective To investigate the effect of dexmedetomidine (DEX) on brain injury and long-term cognitive dysfunction in rats after orthotopic liver transplantation ischemia/reperfusion and its mechanism. Methods Fifty-four male Sprague-Dawley (SD) rats were randomly divided into sham-operated group (n=18), orthotopic liver transplantation ischemia/reperfusion group (I/R group, n=18) and DEX pre-administration group (DEX group, n=18). The orthotopic liver transplantation ischemia/reperfusion models were established in I/R group and DEX group. Rats of the DEX group were intraperitoneally injected with DEX 100 μg/kg 30 min before the incision, and an equal volume of normal saline was injected into rats of the sham-operated group and I/R group at the same time. Twelve rats in each group were sacrificed 3 d after operation and brain tissues were taken. The pathological changes of the hippocampus were observed under light microscope. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) was used to detect neuronal apoptosis index of the hippocampus. Western blotting was used to detect the protein levels of aquaporin 4 (AQP4) and protein kinase C (PKC) in the hippocampus. The remaining 6 rats accepted Morris water maze test to evaluate the long-term cognitive function 30 d after surgery. Results As compared with those in the sham-operated group, rats in the I/R group and DEX group had hippocampus edema and disordered cell arrangement; as compared with sham-operated group, I/R group and DEX group had significantly increased neuronal apoptosis index, significantly increased protein levels of AQP4 and PKC, significantly shorter quadrant retention time of the platform, and statistically longer escape latency (P<0.05). As compared with those in the I/R group, the pathological damage of hippocampal neurons was significantly alleviated, the neuronal apoptosis index was significantly decreased, the protein levels of AQP4 and PKC were statistically decreased, and the residence time of the quadrant in the platform was significantly prolonged, and the escape latency was statistically shorter in DEX group (P<0.05). Conclusion Pre-administration of DEX may reduce the brain damage and improve long-term cognitive dysfunction in rats after orthotopic liver transplantation ischemia/reperfusion, which may be related to down-regulating the AQP4 and PKC expression levels and reducing neuronal apoptosis.

Epidemiological and clinical studies of rotavirus-induced diarrhea in China from 1994-2013.

Group A rotavirus infections cause diarrhea in infants and primarily occur in winter. These viruses are characterized by the viral structural proteins VP7 (G subtypes) and VP4 (P subtypes). Current treatments employ vaccines combined with symptomatic treatments. Through a review of published papers from 2003-2013, which included articles, theses, dissertations and academic conference proceedings in PubMed and CNKI (China National Knowledge Infrastructure), we examined the epidemiological and clinical studies conducted in the 7 regional administrative geographical divisions of the People's Republic of China. Binning of the clinical studies provided a quantitative synthesis of the published incidences of rotavirus-positive numbers, the infection rates and the distributions of different subtypes in the provinces. In an investigation of the 32 provincial administrative regions from 1994-2013, the number of positive infections in children, primarily ≤5 years of age, was highest in Guangdong (187,000 cases), Zhejiang (133,000 cases) and Hubei (11,000 cases) Provinces; the provinces that displayed the highest positive rates were Henan (75.20%), Jilin (64.94%) and the Ningxia Hui Autonomous Region (60.16%). Furthermore, the virus, which causes diarrhea, can also infect adults. Statistics regarding the rotavirus subtypes indicated that the top 3 G-subtypes reported were G3 (8,509 cases), G1 (6,490 cases) and G2 (1,601 cases); the top 3 P-subtypes reported were P[8] (8,483 cases), P[4] (2,017 cases) and P[6] (740 cases). The majority of clinical treatments for these infections comprised a combination of Chinese traditional and Western medicines, together with nursing and the administration of a variety of oral liquids to relieve symptoms. The top 3 positive rates were identified in provinces at higher latitude, which confirms increases in humidity and ambient temperature reduce the rotavirus infection rate and the number of rotavirus infections tends to be highest under cool, dry conditions. However, the presence of rotavirus in low-latitude provinces likely indicates living habits, living environments, and education level influencethe precautions and vaccine utilization rate. The traditional subtypes are the rotavirus G- and P-subtypes, thus allowing for the development of vaccines. In China, the treatments that are primarily used to combat the infection are integrative medicines combining a variety of nursing and adjuvant therapies.