Objective To construct a recombinant expression vector for expression of the function-al domains of dengue virus serotype 1 ( DENV1 ) envelope ( E ) protein in native soluble form. Methods The genes encoding the functional domains of DENV1-E protein (1-394 aa) were amplified with PCR and then cloned into the Psectag2B-Fc eukaryotic expression vector.The 293T cells were transfected with the recombinant vector by cationic lipid-based delivery.The cell clones expressing the fusion DENV1-E-Fc protein were screened out with 2 mg/ml of Zeocin.Immunofluorescence assay ( IFA) was performed to analyze the antigenicity and integrity of the fusion protein.The fusion proteins were purified from cell lysate with Protein-G and further identified by Western blot assay.Results The soluble form of fusion protein with a molecular weight of about 90×103 was obtained at a yield of about 25 μg per 1×107 cells.The results of IFA indicated that the fusion protein kept its integrity with right conformational epitopes.The fusion protein was successfully expressed with the advantage of good specificity as indicated by IFA and Western blot assay. Conclusion The recombinant fusion protein in soluble form was successfully expressed in eukaryotic ex-pression system, which paved the way for further investigation on the function of DENV1 E protein and its protective epitopes.

Objective: To evaluate the intervention effectiveness of co-occurrence and prevention for myopia and obesity among primary and secondary school students, so as to provide a scientific basis for the development of comprehensive intervention measures in myopia and obesity. Methods: From September 2022 to September 2023, a cluster random sampling method was used to select 6 primary schools and 6 junior high schools from Tongzhou District, Beijing. Participants were randomly assigned to an intervention group (914 before intervention and 754 after intervention) and a control group (868 before intervention and 652 after intervention), with an expected duration of one academic year. Based on the RE-AIM framework, integrate resources from families, schools, communities, and medical institutions to develop a school-based intervention technology packagefor the co-occurrence and prevention of myopia and obesity in children. The intervention group received intervention according to the comprehensive intervention technology package, while the control group did not receive any intervention measures. Relevant health indicators during the baseline period and after intervention were measured and collected, and groups were compared by Chi quest test, t-test and Wilcoxon rank sum test. Results: After intervention, the uncorrected visual acuity of primary and secondary school students in the intervention group (4.79±0.30) and the control group (4.77±0.33) both decreased compared to those before intervention (4.80±0.30, 4.90±0.32) ( t =-7.00,-5.24); the decrease in uncorrected visual acuity in the intervention group was smaller than that in the control group( t =5.33)( P <0.01). After intervention, body mass index, waist circumference, hip circumference, and body fat percentage of primary and secondary school students in the intervention group decreased compared to those before intervention. However, the changes in these indicators were not statistically significant ( t/Z =-0.03, - 0.36，- 0.30，- 0.01, P >0.05); the above indicators in the control group increased compared to those before intervention, but only hip circumference and body fat percentage showed statistically significant changes ( t/Z =2.17, 2.62, P <0.05). After intervention, both the intervention group and the control group showed increases in systolic and diastolic blood pressure compared to those before intervention(intervention group: t =2.16,5.29; control group: t =6.84,5.07); the intervention group had lower systolic and diastolic blood pressure than the control group( t = -5.27 , -2.08)( P <0.05). After intervention, the intervention and the control groups had statistically significant differences in cognitive accuracy(92.48%, 69.33%) in terms of "outdoor exercise can prevent myopia" and "having 5 servings of adult fist sized vegetables and fruits every day" ( χ 2=6.30, 7.86, P <0.05). There was a statistically significant difference in the proportion of primary and secondary school students in the intervention group (40.98%) and the control group (35.43%) for "who did not drink sugary drinks for every day in the past 7 days" ( χ 2=4.32, P <0.05). After intervention, the intervention group and the control group showed increases in "school outdoor activity duration on school days" and "outdoor activity duration on rest days" compared to those before intervention ( t/Z =-13.32,-9.71;- 2.59，-2.69）；the behavior rate of "visual acuity measurement frequency at least once every 3 months" in the intervention group (46.68%) and the control group (52.76%) increased compared to those before intervention (36.43%, 44.01%), and the increases in the intervention group were greater than that in the control group ( χ 2=17.52,11.08) ( P <0.05). Conclusions Comprehensive intervention measures have significant intervention effects on controlling the occurrence and development of comorbidity of myopia and obesity in children. It could actively promote collaboration and cooperation among families, schools, communities and medical institutions to reduce the occurrence of myopia and obesity among primary and secondary school students.

BACKGROUNDThe transradial approach is regarded as a useful vascular site for coronary procedures. The aim of this study was to test whether 4Fr catheters assisted by ACIST variable rate injector system can produce comparable angiographic quality and reduce the risk of radial artery injury compared to hand manifold 6 Fr catheters.

METHODSA total of 1816 patients were studied consecutively, among whom 856 patients received coronary angiography by 4 Fr catheters (4Fr group) and 960 patients by 6 Fr catheters (6Fr group). Angiographic and procedural characteristics were observed and recorded. The luminal inner radial arterial diameter before and after the procedure were collected.

RESULTSThe baseline clinical characteristics were similar in both groups. There were no significant differences in procedure time, radiation dose and quality scores in both groups (P > 0.05), but more contrast media was delivered in the 6Fr group (P < 0.001). The mean radial arterial diameter six months after the procedure in the 6Fr group reduced significantly compared to that measured one day prior to the procedure (P < 0.001).

CONCLUSIONSCoronary angiography using the 4Fr catheters with Acist power injection system can achieve an acceptable diagnostic quality while at the same time minimizing radial artery injury and contrast media consumption.

Aged ; Cardiac Catheterization ; instrumentation ; Coronary Angiography ; instrumentation ; methods ; Female ; Humans ; Male ; Middle Aged ; Radial Artery ; diagnostic imaging ; Ultrasonography

Tanshinone Ⅱ A (Tan Ⅱ A) is an extract of traditional Chinese medicine Salvia miltiorrhiza,which can play a natural antioxidant effect.It is mainly used in the treatment of cardiovascular disease.Which has anti-atherosclerosis,reduced myocardial infarct size and improve myocardial oxygen consumption,inhibition of thrombosis,inhibition of platelet aggregation,antitumor activity and other effects,and are mainly used for the treatment of cardiovascular disease in clinical applications.In recent years,a large number of clinical studies and animal experiments have confirmed that tanshinone Ⅱ A in the inhibition of colon cancer cell proliferation,promote cell apoptosis,promote cell invasion and metastasis and other aspects of prominent anti-cancer effect,making it the object of concern to researchers.In this paper,tanshinone ⅡA anti-colon cancer effect of studies were reviewed.

OBJECTIVETo study the changes of vascular endothelial cell function and platelet activation in rabbit spinal cord following ischemia-reperfusion (I/R) injury and their roles in the spinal cord injury.

METHODSRabbit spinal cord I/R injury models were established using Zivin method, and the changes in plasma NO and GMP140 levels were dynamically monitored after the injury.

RESULTSPlasma NO level increased significantly in the I/R group at the end of the ischemia, and reached the peak level at 2 h of reperfusion as compared to that in sham-operated group (P<0.01). Plasma NO level decreased at 6 h of reperfusion, but still significantly higher than the level in the sham-operated group (P<0.05). Plasma GMP140 underwent no significant changes in the sham-operated group, but significantly increased in the I/R group at the end of the ischemia, followed by gradual declination to the normal level at 2 h of reperfusion.

CONCLUSIONSpinal cord I/R injury causes overexpressions of NO and GMP140, suggesting the involvement of endothelial cell injury and platelet overactivation in the pathological process and repair of spinal cord I/R injury.

Animals ; Endothelial Cells ; metabolism ; Nitric Oxide ; blood ; P-Selectin ; blood ; Platelet Activation ; Rabbits ; Reperfusion Injury ; blood ; pathology ; physiopathology ; Spinal Cord ; pathology ; physiopathology

OBJECTIVETo investigate the role of tumor necrosis factor (TNF) alpha on the homing efficiency of hematopoietic stem/progenitor cells (HS/PC) into bone marrow and its mechanism.

METHODSCFSE-labeled umbilical cord blood (UCB) CD34+ cells were transplanted into irradiated (control group) or combined with TNF alpha prepared (experimental group) BALB/c recipient mice. The distribution in peripheral blood, liver, lung and homing characteristics in bone marrow and spleen of UCB CD34+ cells, in BALB/c recipient mice were determined 20 hours after xenotransplantation by flow cytometry (FACS) and their homing efficiency was calculated. ELISA was used to measure serum SDF-1 alpha level. CXCR4 expression levels of on UCB CD34+ cells were assessed by FACS pre-/post-manipulation with TNF alpha. SDF-1 alpha expression level in bone marrow and spleen was tested by immunohistochemistry.

RESULTSUCB CD34+ cells mainly home into recipient mice bone marrow and spleen; The homing efficiency in experimental group bone marrow [(0.65 +/- 0.13)%] was significantly higher than that in control ones [(0.30 +/- 0.09)%, P < 0.01], whereas the homing efficiency in experimental group spleen was dramatically lower than that in control ones (P < 0.01); Treatment with TNF alpha did not affect recipient serum SDF-1 alpha level; After 18 hours co-cultured with TNF alpha, the CXCR4e expression level on UCB CD34+ cells was similar to that on fresh ones; TNF alpha treatment induced significantly higher SDF-1 alpha expression on osteoblastic and stromal cells in bone marrow, and reversed spleen SDF-1 alpha gradient that was originally favorable for CD34+ cells homing.

CONCLUSIONTNF alpha enhances the homing efficiency of HS/PC via up-regulating SDF-1 alpha gradient in bone marrow, and might be an useful enhancer for HS/PC homing in clinical practice.

Animals ; Antigens, CD34 ; Bone Marrow ; Cell Movement ; Cell Separation ; Chemokine CXCL12 ; metabolism ; Female ; Fetal Blood ; cytology ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; drug effects ; immunology ; metabolism ; Humans ; Mice ; Mice, Inbred BALB C ; Receptors, CXCR4 ; metabolism ; Transplantation Conditioning ; Transplantation, Heterologous ; Tumor Necrosis Factor-alpha ; pharmacology

Aspergillus fumigatus ; Cytokines ; Humans ; Interleukin-1beta ; Keratitis ; Receptors, IgE ; Tumor Necrosis Factor-alpha ; Tumor Necrosis Factors

Objective:To explore the value of the static and dynamic pelvic floor MRI and diffusion tensor imaging (DTI) in evaluating the morphology and function of urethra in patients with stress urinary incontinence (SUI).Methods:From July 2020 to February 2021, a total of 28 patients with SUI and 45 age-matched healthy controls were prospectively collected at the First Affiliated Hospital of Zhengzhou University. The static and dynamic pelvic floor MRI and DTI were performed for all subjects. The thickness of internal and external sphincter of middle urethra were measured on static MRI images. The functional urethral length (FUL) was measured both on static and maximal strain phase of dynamic MRI images, then the difference of FUL was calculated. The presence of bladder neck funneling and urethra opening were observed on static and dynamic MRI. The muscle fiber bundle image of urethral sphincter complex was obtained by post-processing of DTI original images. The anisotropy fraction (FA), apparent diffusion coefficient (ADC) and three eigenvalues (λ1, λ2, λ3) of annular sphincter and central longitudinal muscle in middle urethra were measured. The independent sample t test and chi-square test were used to analyse the difference of measured parameters in MRI, parameters of DTI and imaging signs between the two groups. Results:Compared with healthy controls, the SUI patients showed that the thickness of external sphincter in middle urethral and FUL in static status and maximal strain phase were significantly decreased ( t=-3.95, -5.72, -8.41, all P<0.001), the difference of FUL between static status and maximal strain phase was significantly increased ( t=4.41, P<0.001). The positive rate of bladder neck funneling in static status and maximal strain phase, urethral opening in maximal strain phase of SUI group increased significantly (χ2=23.09 , 22.25, 26.59, all P<0.001). In SUI group, the FA value of middle urethral annular sphincter decreased significantly ( t=-3.48, P=0.001), while the ADC, λ2 and λ3 values increased significantly ( t=3.19, 2.15 , 2.06, and P=0.002, 0.038 , 0.046, respectively). There was no significant difference in DTI parameters of middle urethral longitudinal muscle between the two groups (all P>0.05). Conclusions:Static and dynamic MRI and DTI techniques can objectively evaluate the changes of urethral morphology and function of SUI patients. The thinning of the external sphincter in the middle urethra, shortening of the FUL and the destruction of the microstructure of the annular sphincter fiber bundle were the main alterations of SUI patients.

Objective:To investigate the effects of Ziyin Liangxue formula combined with prednisone on immune function and the ST2/IL-33 pathway in mice with immune thrombocytopenia.Methods:In 40 BALB/c mice,32 were constructed as immune thrombocytopenia mouse models by antiplatelet serum injection.After successful modeling,the mice were randomly divided into model group,Ziyin Liangxue formula group(0.2 ml/10 g),prednisone group(0.2 ml/10 g),and Ziyin Liangxue formula+prednisone group(0.2 ml/10 g),8 mice in each group,and the other 8 mice were set as control group.The drugs were administered by gavage at the dose,and the model group and control group were given equal amounts of saline by gavage once a day for 2 weeks of continuous intervention.Blood samples and spleen tissues were collected,the peripheral platelet count was measured by automatic hematology analyzer,the pathological changes in spleen tissue was observed by HE staining,the levels of serum transforming growth factor(TGF)-β,interleukin(IL)-17,and peripheral blood thrombopoietin(TPO)were detected by enzyme-linked immunosorbent assay(ELISA),the expression of IL-33,sST2,and ST2 in spleen tissue was detected by Western blot,and the cell counts of peripheral blood Th17 and Treg were detected by flow cytometry.Results:Compared with the control group,the number of platelets,the level of TPO,TGF-β,and Treg cells were significantly decreased(P＜().05),while the level of IL-17,Thl7 cells,and the expression of IL-33,sST2,and ST2 protein were significantly increased in the model group(P＜0.01).Compared with the model group,the number of platelets,the level of TPO,TGF-β,and Treg cells were significantly increased(P＜0.05),while the level of IL-17,Th17 cells,and the expression of IL-33,sST2,and ST2 protein were significantly decreased in the Ziyin Liangxue formula+prednisone group(P＜0.01).Conclusion:Ziyin Liangxue formula+prednisone can effectively regulate Th17/Treg balance,thus effectively improve immune thrombocytopenia,and the mechanism may be related to the regulation of ST2/IL-33 signaling pathway.

Interferon-γ (IFN-γ) has been used to control cancers in clinical treatment. However, an increasing number of reports have suggested that in some cases effectiveness declines after a long treatment period, the reason being unclear. We have reported previously that long-term IFN-γ treatment induces malignant transformation of healthy lactating bovine mammary epithelial cells (BMECs) in vitro. In this study, we investigated the mechanisms underlying the malignant proliferation of BMECs under IFN-γ treatment. The primary BMECs used in this study were stimulated by IFN-γ (10 ng/mL) for a long term to promote malignancy. We observed that IFN-γ could promote malignant cell proliferation, increase the expression of cyclin D1/cyclin-dependent kinase 4 (CDK4), decrease the expression of p21, and upregulate the expression of cellular-abelsongene (c-Abl) and histone deacetylase 2 (HDAC2). The HDAC2 inhibitor, valproate (VPA) and the c-Abl inhibitor, imatinib, lowered the expression level of cyclin D1/CDK4, and increased the expression level of p21, leading to an inhibitory effect on IFN-γ-induced malignant cell growth. When c-Abl was downregulated, the HDAC2 level was also decreased by promoted proteasome degradation. These data suggest that IFN-γ promotes the growth of malignant BMECs through the c-Abl/HDAC2 signaling pathway. Our findings suggest that long-term application of IFN-γ may be closely associated with the promotion of cell growth and even the carcinogenesis of breast cancer.
Animals ; Carcinogenesis/pathology* ; Cattle ; Cell Cycle Proteins/metabolism* ; Cell Proliferation/drug effects* ; Cell Transformation, Neoplastic/pathology* ; Cells, Cultured ; Epithelial Cells/pathology* ; Female ; Histone Deacetylase 2/metabolism* ; Imatinib Mesylate/pharmacology* ; Interferon-gamma/pharmacology* ; Mammary Glands, Animal/pathology* ; Mammary Neoplasms, Experimental/pathology* ; Proto-Oncogene Proteins c-abl/metabolism* ; Signal Transduction ; Valproic Acid/pharmacology*