Adolescent ; Adult ; Blood Pressure ; Gravity Suits ; Heart Function Tests ; Humans ; Male ; Positive-Pressure Respiration ; Young Adult

To understand the structure of the soluble region of Enterovirus 68 3A protein, we construct a prokaryotic expression vector expressing the soluble region of EV-D68 3A protein, and identify the forms of expression product after purification. The EV-D68 3A(1-61) gene was amplified by PCR and then cloned into the expression vector pET-28a-His-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 induced by IPTG to express the fusion protein His-SUMO-3A(1-61). The recombinant protein was purified by Ni-NTA Agarose and cleaved by ULP Protease to remove His-SUMO tag. After that, the target protein 3A(1-61) was purified by a series of purification methods such as Ni-NTA, anion exchange chromatography and gel filtration chromato- graphy. Chemical cross-linking reaction assay was taken to determine the multiple polymerization state of the 3A soluble region. A prokaryotic expression vector pET28a-His-SUMO-3A(1-61) expressing the solution region of EV-D68 3A was successfully constructed and plenty of highly pure target proteins were obtained by multiple purification steps . The total protein amount was about 5 mg obtained from 1L Escherichia coli BL21 with purity > 95%. At the same time, those results determined the homomultimer form of soluble 3A construct. These data demonstrated that the expression and purification system of the soluble region of 3A were successfully set up and provide some basic konwledge for the research about 3A crystal structure and the development of antiviral drugs targeted at 3A to block viral replication.
Amino Acid Sequence ; Enterovirus D, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; metabolism

Objective To investigate radionuclide imaging and routine CT in diagnosing hepatic focal nodular hyperplasia (FNH) and the combined diagnostic value of the two modalities. Methods Thirty-two patients with hepatic FNH were retrospectively studied. All patients underwent routine CT scan. Twenty-four patients were examined by 99Tcm-sulfur colloid (SC) hepatic planar scintigraphy and SPECT/CT imaging, and then patients who had abnormal foci underwent 99Tcm-diethyl iminodiacetic acid (EHIDA) triple-phase hepatobiliary imaging. x2 -test of four-table or Fisher exact probabilities in 2 × 2 table was applied for statistical analysis. Results Of all 32 patients pathologically diagnosed as FNH with single solitary nodule, 25 were classified as classic type and the rest 7 as non-classic type. Although routine CT found all hepatic lesions, only 15 cases were diagnosed pathologically as FNH classic type but the rest were either misdiagnosed or left as indeterminate. On radionuclide imaging (hepatic colloid scintigraphy plus triple-phase hepatobiliary images), 11 patients with big foci (with maximal diameter ＞3 cm) out of 24 patients were correctly diagnosed as FNH, with 7 diagnosed as classic type FNH and 4 as non-classic. Other 13 patients were either misdiagnosed or simply missed. The diagnosing rates of routine CT and radionuclide imaging were60.0% (15/25) and 38.9% (7/18) for FNH classic type, 0/7 and 4/6 for non-classic type,50.0% (10/20) and 73.3% (11/15) for big foci, 41.7% (5/12) and 0/9 forsmall foci (with maximal diameter≤3 cm), respectively. The total diagnosing rate of radionuclide imaging combined with routine CT was significantly higher than that of routine CT or radionuclide imaging alone ( x2 = 4. 48, P ＜ 0. 05;x2 =4.27, P ＜0.05 ). Conclusion Radionuclide imaging in combination with routine CT may improve the diagnostic accuracy for hepatic FNH patients.

Objective To investigate the protective effect of theaflavins on thymus injury caused by total body irradiation (TBI).Methods Twenty-five C57BL/6 mice were randomly divided into 5 groups:control group,4 Gy TBI group,4 Gy TBI + 25 mg/kg theaflavins group,4 Gy TBI + 50 mg/kg theaflavins group and 4 Gy TBI + 100 mg/kg theaflavins group.Thymus index and total number of thymocytes were detected at the 14th d post-irradiation to determine the optimal dose of theaflavins.According to this optimal dose,32 C57BL/6 mice were randomly divided into 4 groups:control group,theaflavins group,4 Gy TBI group and 4 Gy TBI + theaflavins group.Thymus histomorphology,CD4CD8 T cell subsets,and reactive oxygen species (ROS) in thymocytes were examined at the 14th d post-irradiation.Results The irradiated thymus exhibited decreased thymus index and total number of thymocytes (P ＜ 0.05),aberrant histomorphology and T cell subsets (P ＜ 0.05),and increased ROS level in thymocytes (P ＜ 0.05).Compared with 4 Gy TBI group,the thymus index and total number of thymocytes were significantly increased in 4 Gy TBI + 50 mg/kg theaflavins group (P ＜ 0.05).The total number of thymocytes was significantly higher in 4 Gy TBI + 50 mg/kg theaflavins group than that in 4 Gy TBI + 25 mg/kg theaflavins group (P ＜ 0.05).Therefore,50 mg/kg theaflavins was chosen as the optimal dose for subsequent experiments.Moreover,the aberrant histomorphology of irradiated thymus was alleviated by theaflavins.A decline in the percentage of CD4-CD8-T cells and an elevation of CD4+CD8-and CD4+CD8+ T cells were found in irradiated mice administered with theaflavins (P ＜ 0.05).Compared with 4 Gy TBI group,the ROS level was significantly decreased in 4 Gy TBI + theaflavins group (P ＜ 0.05).Conclusion Theaflavins exhibits a protective effect on radiation-induced thymus injury.

Objective To report the integrated progress of the hydatid disease information management system,and to pro-vide the reference for further system improvements by analysis of results on simulation test feedback. Methods The work of in-stitutional code matching by collecting fundamental and integrated information of the system in epidemic areas of hydatid disease was carried out,and professional control agencies were selected to carry out the simulation test. Results The results of agen-cies code matching at stage indicated the average completion rate was 94.30% on administrative agencies,69.94% on registered professional agencies and 56.40% on professional institutions matching related to hydatid disease prevention and control imple-ments in seven provinces(autonomous regions)and Xinjiang Production and Construction Corps. Meanwhile,the response rate of open-ended proposals was 93.33% on fifteen feedbacks,and the statistics showed 21.43% believed the system was low fluen-cy,64.29% considered the system was inconvenience for data inputs and 42.86% considered it would be improved on system sta-tistics functions,of which 27.78% were provincial users,22.22% were the city users and 50.00% were the county users. Con-clusion The hydatid disease prevention information management system meets the fundamental needs of the majority agencies in hyperendemic areas of echinococcosis,it needs to develop the further test with more agencies joining after the work of the in-stitutional code matching completion and the system service improvement in the next stage.

Radiotherapy is one of major cancer treatment methods.However,radiation resistance is an important reason to restrict the efficacy of radiotherapy and lead to treatment failure.In recent years,the relationship between the canonical Wnt signaling pathway and tumor radiation resistance has more and more attention of the scholars.This review summarized recent ten years findings concerning the canonical Wnt signaling pathway and tumor radiation resistance and tried to find some valuable rules or some internal relationships among different pathways by systemically analyzing.

Objective To study the interobserver variabilities and the differential diagnosis value of Breast Imaging Reporting and Data System-Ultrasound (BI-RADS-US) lexicon for small ( ≤ 2 cm) breast nodules. Methods Between January 2009 and December 2011, 289 patients with small (≤2 cm) breast nodules (n=317) were included. According to sizes, the lesions were divided into two groups, i.e., 0-1 cm (n=160) group and 1-2 cm (n=157)group. Each lesion was described independently by 3 radiologists using BI-RADS-US lexicon. Interobserver variabilities were assessed by Kappa test. Chi-square test was used to compare the frequency difference of the descriptors between malignant and benign lesions. Sensitivity, speciifcity, accuracy, positive predictive value and negtive predictive value were calculated. Results (1)Moderate agreements were obtained for lesion shape, orientation, margin, echo pattern, surrounding tissue and calciifcations (κ=0.44, 0.57, 0.48, 0.43, 0.51 and 0.57) in 0-1 cm group. Substantial agreements were obtained for lesion shape, orientation, margin and echo pattern (κ=0.65, 0.61, 0.64 and 0.63) in 1-2 cm group. (2)Irregular shape, non-parallel orientation, non-circumscribed margin, echogenic halo and microcalciifcations were more frequently found in malignant nodules than in benign nodules in 0-1 cm group [52.3% (34/65) vs 20.0% (19/95), 38.5%(25/65) vs 13.7%(13/95), 75.4%(49/65) vs 32.6%(31/95), 18.6%(12/65) vs 0 (0/95) and 10.8%(7/65) vs 2.1%(2/95);χ2=18.19, 13.08, 28.22, 16.39 and 3.95;P=0.000, 0.000, 0.000, 0.000 and 0.047]. Similarly, irregular shape, non-parallel orientation, non-circumscribed margin, echogenic halo, shadowing, changes of Cooper′s ligament and microcalciifcations were signiifcantly more frequent found in malignant nodules than in benign nodules in 1-2 cm group [74.2%(49/66) vs 12.1%(11/91), 36.3%(24/66) vs 5.5%(5/91), 93.9%(62/66) vs 22.0%(20/91), 37.9%(25/66) vs 3.3%(3/91), 30.3%(20/66) vs 7.7%(7/91), 15.2%(10/66) vs 0 (0/91) and 16.7%(11/66) vs 4.4%(4/91);χ2=62.59, 24.21, 79.40, 31.22, 13.73, 12.30 and 6.67;P=0.000, 0.000, 0.000, 0.000, 0.000, 0.000 and 0.010]. (3)In both groups, a good sensitivity was demonstrated (75.4%&93.9%) when using the non-circumscribed margin as a criterion for malignancy, and high speciifcity was achieved in two groups (80.0%-100%and 87.9%-100%) when other descriptors including irregular shape, non-parallel orientation, echogenic halo, shadowing, changes of Cooper′s ligament and microcalciifcations were used as differentiation criteria. Conclusions Good interobserver agreement can be achieved using the BI-RADS-US lexicon in the diagnosis of small breast nodules. Non-circumscribed margin are proved as the most valuable sign for screening malignant breast lesions ≤ 2 cm. High speciifcity was found for irregular shape, nonparallel orientation, echogenic halo, shadowing, Cooper′s ligament changes and microcalciifcations, which can help biopsy and preoperative diagnosis.

Phospholipase D(PLD) is ubiquitous in bacteria,fungi,and mammal.In pathogenic microorganisms,PLD can be pathogenic determinant and play a role in spore generation.In mammalian cells,PLD functions in several signal transduction pathways,such as membrane transportation,mitosis regulation,and actin cytoskeleton regulation.In the process of pathogens invasion host cells,both of the pathogen and host cells’ PLD will be activated and a series of cascade reaction will be generated.During this process,pathogen’s PLD can regulate the polymerization and reorganization of its own actin filaments and induce the polymerization or reorganization of the host cell actin filaments near the foci,thus to promote the phagocytosis of the pathogen by host cell.Investigating the role of PLD activation in the infection will be significance for further understanding the molecular mechanism of pathogen-host cell interaction.

This paper investigates the effect of myricetin (MYR) on renal fibrosis induced by unilateral ureteral obstruction (UUO) and common bile duct ligation (CBDL) in mice and its mechanism. The animal experiment has been approved by the Ethics Committee of China Pharmaceutical University (NO: 2022-10-020). Thirty-five ICR mice were divided into control, UUO, UUO+MYR, CBDL and CBDL+MYR groups. H&E and Masson staining were used to detect pathological changes in kidney tissues. Western blot (WB) was used to detect the expression of fibrosis-related proteins in renal tissue, and total superoxide dismutase (SOD) activity detection kit (WST-8) was used to detect the changes of total SOD in renal tissue of CBDL mice. In vitro, HK-2 cells and transforming growth factor beta 1 (TGF-β1, 10 ng·mL^-1) were used to induce fibrotic model, and high glucose (30 mmol·L^-1) was used to induce oxidative stress model, and then treated with different concentrations of MYR, WB was used to detect the expression of fibrosis and oxidative stress-related proteins, while NIH/3T3 cells were treated with different concentrations of MYR, and their effects on cell proliferation were detected by 5-bromo-2′-deoxyuridine (Brdu). The results showed that the renal lesions in UUO group and CBDL group were severe, collagen deposition was obvious, the expression of collagen-Ⅰ (COL-Ⅰ), α-smooth muscle actin (α-SMA), fibronectin (FN), vimentin and plasminogen activator inhibitor-1 (PAI-1) protein was up-regulated, and the activity of SOD enzyme in CBDL group was significantly decreased. MYR partly reversed the above changes after treatment. MYR inhibited the proliferation of NIH/3T3 cells but had no effect on the proliferation of HK-2 cells, and decreased the upregulation of PAI-1, FN and vimentin in HK-2 cells stimulated by TGF-β1. MYR can also up-regulate the down-regulation of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in HK-2 cells stimulated by high glucose. To sum up, MYR can improve renal fibrosis in vivo and in vitro, probably by inhibiting the proliferation of fibroblasts and activating Nrf2/HO-1 signal pathway to inhibit oxidative stress.

Applications of network pharmacology are increasingly widespread and methods abound in the field of drug development and pharmacological research. In this study, we choose rosiglitazone compound as the object to predict the targets and to discuss the mechanism based on three kinds of prediction methods of network pharmacology. Comparison of the prediction result has identified that the three kinds of prediction methods had their own characteristics: targets and pathways predicted were not in accordance with each other. However, the calcium signaling pathway could be predicted in the three kinds of methods, which associated with diabetes and cognitive impairment caused by diabetes by bioinformatics analysis. The above conclusion indicates that the calcium signaling pathway is important in signal pathway regulation of rosiglitazone compound, which provides a clue to further explain the mechanism of the compound and also provides a reference for the selection and application of methods of network pharmacology in the actual research.
Calcium Signaling ; Cognitive Dysfunction ; Computational Biology ; Diabetes Mellitus ; Humans ; Pharmacology ; methods ; Thiazolidinediones ; pharmacology