Robust humoral and cellular immune responses induced by novel DNA vaccine against West Nile virusin in mice / 中华微生物学和免疫学杂志

Objective:To evaluate the immunological efficacy of a novel DNA vaccine against West Nile virus (WNV) in a mouse model.Methods:A DNA vaccine VRC-prME expressing the precursor membrane (prM) and envelope protein (E) of WNV Xinjiang strain (XJ11129-3) was constructed and its ability to express virus-like particles was verified in vitro. C57BL/6 mice were immunized twice with VRC-prME via intramuscular injection combined with electroporation with an interval of four weeks. Enzyme-linked immunoassay (ELISA) was used to detect serum antibodies after immunization. WNV (NY99 strain) single-round infectious particles were used to detect neutralizing antibodies. Cellular immune responses were analyzed by enzyme-linked immunoblot assay (ELISPOT) and intracellular cytokine staining (ICS). Results:VRC-prME induced a strong Th1-biased antibody response in mice that could cross-neutralize the WNV (NY99 strain) single-round infectious particles two weeks after the boost immunization. Moreover, the vaccine also elicited antigen-specific multifunctional CD8 + T cell responses (IFN-γ, IL-2, TNF-α). Conclusions:The novel DNA vaccine prepared in this study, expressing the prME protein of WNV XJ11129-3 strain, could induce stronger humoral and cellular immune responses in mice, which was worthy of further research and development for the prevention of WNV infection in China.

Sesamin and vitamin E ameliorate aortic endothelial dysfunction rats induced by D-galactose and aluminum trichloride via regula?tion of endothelial nitric oxide synthase and NAD(P)H oxidase 4 / 中国药理学与毒理学杂志

OBJECTIVE To observe the protective effect of sesamin (Ses) and vitamin E (Vit E) against aortic endothelial dysfunction in rats induced by D-galactose (D-gal) and aluminum trichloride (AlCl3), and explore its conceivable mechanisms. METHODS A model of aortic endothelial dysfunction rats was established by D-gal (180 mg · kg-1, ip) combined with AlCl3 (15 mg · kg-1, ig) for 84 d. Model rats were randomly divided into model, model+Vit E 10 mg·kg-1, model+Ses 160 mg·kg-1, and model+Ses 160 mg · kg-1+Vit E 10 mg · kg-1 groups. After 70 d of treatment with Ses and Vit E, systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean blood pressure (MBP) were measured by tail cuff. The rats were anesthetized by sodium pentobarbital (30 mg·kg-1, ip). Thoracic aortas from the rats were removed and divided into two parts (3 mm in length). The relaxation of the aortic ring induced by acetylcholine (ACh) and sodium nitroprusside was measured. The primary pathologic changes in the aorta were observed by HE staining. Total antioxidant capacity (T-AOC), hydrogen peroxide (H2O2) and nitric oxide (NO) in serum were measured by colorimetric analysis. The expression of endothelial nitric oxide synthase (eNOS) positive cells in the aorta were measured by immunohistochemistry. The expres?sions of eNOS and NAD(P)H oxidase 4 (NOX4) protein in the aortal were detected by Western blotting. RESULTS Compared with the model group, the relaxation response with increase in ACh concentra?tion (1×10-7-1×10-4 mol·L-1) was enhanced (P<0.01) in model+Ses+Vit E, SBP, DBP and MBP decreased (P<0.01), the serum T-AOC and NO level were increased (P<0.01), the serum H2O2 levels were reduced (P<0.01), the eNOS expression was increased (P<0.01) and NOX4 expression was reduced (P<0.01) in each treatment group. Compared with model+Ses, the SBP, DBP and MBP were lower (P<0.01 or P<0.05), the serum H2O2 level was lower (P<0.01), the serum NO level was increased (P<0.05), the eNOS expression level was higher (P<0.01) and the NOX4 expression level was reduced (P<0.05) in model+Ses+Vit E. Compared with the model+Vit E, the serum T-AOC and NO levels were increased (P<0.05), the serum H2O2 level was lower (P<0.01), eNOS expression was increased (P<0.01) and NOX4 expression was reduced (P<0.05) in model+Ses+Vit E group. CONCLUSION Ses and Vit E can ameliorate aortic endothelial dysfunction of rats induced by D-gal and AlCl3 via the regulation of eNOS and NOX4.

Effect of salvianolic acid B on high glucose-induced phenotypic transition and extracellular matrix secretion in human glomerular mesangial cells / 中国病理生理杂志

AIM:To investigate the effect of salvianolic acid B (Sal B) on high glucose-induced phenotypic transition and extracellular matrix (ECM) secretion in human glomerular mesangial cells (HGMCs) and the underlying mechanisms.METHODS:HGMCs were randomly divided into control group, high glucose group and high glucose plus high dose, medium dose and low dose of Sal B groups.The HGMCs except those in control group were exposed to high glucose (33.3 mmol/L) for 72 h, while those in Sal B groups were co-incubated with indicated concentrations of Sal B.The protein levels ofα-smooth muscle actin (α-SMA) , transforming growth () and phosphorylated Smad2 and p38 mitogen-activated protein kinase (MAPK) were determined by Western blot.The secretion levels of collagen type I (Col I) , collagen type III (Col III) , fibronectin (FN) and laminin (LN) were measured by ELISA.RESULTS:Exposure to high glucose markedly increased the protein expression ofI, Col III, FN and LN in the HG-MCs (P<0.01).The phosphorylation levels of Smad2 and p38 MAPK were also significantly increased (P<0.01).Coincubation with Sal B evidently decreased the protein expression ofI, Col III, FN and LN in the HGMCs induced by high glucose (P<0.05 or P<0.01).The phosphorylated levels of Smad2 and p38 MAPK were also reduced noticeably (P<0.05 or P<0.01).CONCLUSION:Sal B significantly suppresses high glucose-induced phenotypic transition and ECM secretion in the HGMCs, which might be attributed, at least partly, to inhibition ofSmad signaling pathway and p38 MAPK activation.

Establishment and evaluation of receptor binding domain (RBD)-based ELISA for Middle East respiratory syndrome coronavirus (MERS-CoV) antibody detection / 中华微生物学和免疫学杂志

Objective@#To establish an indirect enzyme-linked immunosorbent assay (ELISA)and to compare the efficiency of receptor binding domain (RBD) proteins in different forms for Middle East respiratory syndrome coronavirus (MERS-CoV) antibody detection.@*Methods@#The monomeric and trimeric forms of MERS-CoV RBD were expressed in Bac-insect cells, 293T cells and ExpiCHO-S™ expression system and then purified. The purified RBD proteins were identified with native gel electrophoresis and Western blot. Then, an equal amount of each RBD protein was used as coating antigen to establish an ELISA for detecting MERS-CoV IgG titer. For comparison, the newly developed ELISA and the commercial MERS-CoV IgG antibody detection kit (Euroimmune with S1 as the coating antigen) were used to measure the MERS-CoV antibody reference panel supplied by World Health Organization (WHO).@*Results@#The purified monomeric and trimeric MERS-CoV RBD were successfully prepared using 293T cells and ExpiCHO-S™ system. RBD antigens of different forms and from different systems could recognize MERS-CoV specific antibody without having any cross reaction with the sera from healthy adults. The in-house RBD-based ELISA had good detection consistency with the Euroimmune commercial kit. The positive samples showed higher and more concentrated values based on the RBD trimer than the monomer.@*Conclusions@#Novel indirect ELISA methods based on the monomeric and trimeric forms of RBD protein were established. The trimetric form-based ELISA achieved higher detection efficiency than the one using the monomer antigen, suggesting that it could be uses as a competent alternative to the commercial kit.

The Lymphatic Drainage Pattern of Internal Mammary Sentinel Lymph Node Identified by Small Particle Radiotracer ((99m)Tc-Dextran 40) in Breast / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The purpose of this study was to detect the lymphatic drainage pattern of internal mammary area and verify the concept of internal mammary sentinel lymph node (IM-SLN) in breast. MATERIALS AND METHODS: A small particle radiotracer ((99m)Tc-Dextran 40) was prepared and tested. (99m)Tc-Dextran 40 was injected into intraparenchyma at the sound breast by a modified radiotracer injection technique. Subsequently, dynamic single-photon emission computed tomography (SPECT), computed tomography (CT), and SPECT/CT combination images were performed to identify the radioactive lymph vessels and internal mammary lymph nodes (IMLNs). The direction of lymph drainage and the location of the IMLNs were identified in the SPECT/CT imaging. RESULTS: The radiochemical purity of (99m)Tc-Dextran 40 was > 95%. (99m)Tc-Dextran 40 could drainage into first, second, and third lymph node and the radioactive lymph node could be detected by the γ detector in the animal experiment. After (99m)Tc-Dextran 40 injecting into intraparenchyma, 50.0% cases (15/30) were identified the drainage lymphatic vessels and radioactive IMLNs by SPECT. The drainage lymphatic vessel was found from injection point to the first IMLN (IM-SLN) after 10.5±0.35 minutes radiotracer injection, and then (99m)Tc-Dextran 40 was accumulated into the IM-SLN. The combination imaging of SPECT/CT showed the second IMLN received the lymph drainage from the IM-SLN. The lymphatic drainage was step by step in the internal mammary area. CONCLUSION: The lymph was identified to drain from different regions of the breast to IM-SLN, and then outward from IM-SLN to other IMLN consecutively. It demonstrated the concept of the IM-SLN and provided more evidences for the application of internal mammary sentinel lymph node biopsy.

Establishment and evaluation of receptor binding domain (RBD)-based ELISA for Middle East re-spiratory syndrome coronavirus (MERS-CoV) antibody detection / 中华微生物学和免疫学杂志

Objective To establish an indirect enzyme-linked immunosorbent assay (ELISA)and to compare the efficiency of receptor binding domain (RBD) proteins in different forms for Middle East re-spiratory syndrome coronavirus (MERS-CoV) antibody detection. Methods The monomeric and trimeric forms of MERS-CoV RBD were expressed in Bac-insect cells, 293T cells and ExpiCHO-STM expression sys-tem and then purified. The purified RBD proteins were identified with native gel electrophoresis and Western blot. Then, an equal amount of each RBD protein was used as coating antigen to establish an ELISA for de-tecting MERS-CoV IgG titer. For comparison, the newly developed ELISA and the commercial MERS-CoV IgG antibody detection kit (Euroimmune with S1 as the coating antigen) were used to measure the MERS-CoV antibody reference panel supplied by World Health Organization (WHO). Results The purified mon-omeric and trimeric MERS-CoV RBD were successfully prepared using 293T cells and ExpiCHO-STM system. RBD antigens of different forms and from different systems could recognize MERS-CoV specific antibody without having any cross reaction with the sera from healthy adults. The in-house RBD-based ELISA had good detection consistency with the Euroimmune commercial kit. The positive samples showed higher and more concentrated values based on the RBD trimer than the monomer. Conclusions Novel indirect ELISA methods based on the monomeric and trimeric forms of RBD protein were established. The trimetric form-based ELISA achieved higher detection efficiency than the one using the monomer antigen, suggesting that it could be uses as a competent alternative to the commercial kit.

Neutralizing Antibody Titer Test of Ebola Recombinant Protein Vaccine and Gene Vector Vaccine pVR-GP-FC / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>In previous studies, we immunized mice with Ebola recombinant protein vaccine and gene vector vaccine. Both stimulated high levels of humoral immunity. In this work, we constructed a pseudovirus containing Ebola membrane proteins to verify whether the two immunization strategies can induce neutralizing antibodies in mice.</p><p><b>METHODS</b>A pseudovirus containing an Ebola virus membrane protein based on the HIV-1 viral gene sequence was constructed and evaluated using a known neutralizing antibody. The titer of the neutralizing antibody in the sera of mice immunized with the recombinant protein and the gene vector vaccine was examined using a neutralization test.</p><p><b>RESULTS</b>Ebola pseudovirus was successfully prepared and applied for neutralizing antibody detection. Immunological experiments showed that recombinant protein GP-Fc and gene vaccine pVR-modGP-Fc had good immunogenicity. The titer of the bound antibody in the serum after 8 weeks of immunization in mice was more than 1:105, and the recombinant protein induced greater humoral immunity. The results of the neutralization test based on the Ebola pseudovirus system demonstrated that both vaccines induced production of protective antibodies, while the gene vaccine induced a higher titer of neutralizing antibodies.</p><p><b>CONCLUSION</b>An Ebola pseudovirus detection system was successfully established and used to evaluate two Ebola vaccines. Both produced good immunogenicity. The findings lay the foundation for the development of new Ebola vaccines and screening for neutralizing monoclonal antibodies.</p>

Prevalence of depressive symptoms among adolescents / 预防医学

Objective To evaluate the prevalence of depressive symptoms and influencing factors among adolescents. Methods A total of 635 students aged 13 to 18 years were selected in March 2017 and were investigated with general information questionnaire, CES-D and CTQ-SF. Results 630 questionnaires are effective and the positive rate of depression was 29.84％(188/630) . The average score of CTQ was 32.15±2.98; 38.73％ of the students were disregarded and only 2.22％ of that were abused during their childhood. 40.00％ of the students had no CTQ, 1 kinds of CTQ accounted for 31.43％ , 2 kinds of CTQ accounted for 18.25％, and more than 3 CTQ accounted for 10.32％. Multivariate logistic regression analysis show that gender (OR=1.034, 95％ CI: 1.012-1.056) , parents' marital status (OR=1.124, 95％ CI: 1.087-1.162) , family atmosphere (OR=1.025, 95％ CI: 1.024-1.158) , CTQ cumulative number (ORCTQ=1=1.528, 95％ CI: 1.214-1.923; ORCTQ=2=3.067, 95％ CI: 1.325-7.102; ORCTQ≥3=10.361, 95％ CI: 3.059-35.093) were the risk factors for depression. Conclusion Gender, parents' marital status, family atmosphere and CTQ cumulative number were risk factors for depression in adolescents.

Development of the Occupational Stressor Scale for Chinese Seafarers (OSSCS) / 中国心理卫生杂志

Objective:To develop a standardized scale for measuring Chinese seafarers' occupational stressor and test its reliability and validity.Methods:The scale was developed based onliterature research,open questionnaire investigation,interviews and expert discussion.531 seafarers from nationwide were recruited to complete the pre-investigation.Totally 422 seafarers were tested with formal version for further confirmatory factor analysis and internal consistency reliability.Forty-eight seafarers were tested for test-retest reliability.The criterion validity was tested with the Symptom Checklist 90 (SCL-90).Results:The Occupational Stressor Scale for Chinese Seafarers (OSSCS) included 52 items.Exploratory factor analysis generated 6 factors as occupational environment,health,interpersonal relationships,social environment,love and marriage,career development,which accounted for 62.0％ of the variance in all.Confirmatory factor analysisshowed that the model fit well (x2/df =2.28,CFI =0.94,IFI =0.93,NFI =0.91,TLI =0.92,RMSEA =0.06).The OSSCS scores were positively correlated with SCL-90 scores (r =0.37-0.67,Ps ＜ 0.01).Internal consistency reliabilities were 0.73-0.97,while test-retest reliabilities was 0.82-0.91.Conclusion:It indicates the developed Occupational Stressor Scale for Chinese Seafarers (OSSCS) is with good validity and reliability.

Effect of Point Neiguan(PC6) Electroacupuncture Pretreatment on NO, NOS and Mitochondrial Membrane Potential in Rats with Myocardial Ischemia/Reperfusion Injury / 上海针灸杂志

Objective To explore the effect of point Neiguan(PC6) electroacupuncture pretreatment on nitric oxide (NO), nitric oxide synthase (NOS) and mitochondrial membrane potential by determining NO, NOS and mitochondrial membrane potential in rats with myocardial ischemia/reperfusion injury (MIRI).Method Forty male SD rats were randomized to sham operation, ischemia/reperfusion model, point Neiguan electroacupuncture and point Huantiao(GB30) electroacupuncture groups, 10 rats each. The model was made by coronary artery ligation. Before model making, electroacupuncture was given to the point Neiguan electroacupuncture and point Huantiao electroacupuncture groups, 20 min/d for a total of 7 d. T wave value in ECG leadⅡ was measured before and after model making. Myocardial pathomorphological changes were examined by HE staining. Serum NO and NOS contents were measured by a colorimetric nitrate reductase assay. Cardiomyocyte mitochondrial membrane potential was determined by fluorescence techniques.Result Serum NO and NOS contents and mitochondrial membrane potential decreased significantly in the model group compared with the sham operation group (P<0.05). Serum NO and NOS contents increased significantly in the point Neiguan electroacupuncture group compared with the model, sham operation and point Huantiao electroacupuncture groups (P<0.01,P<0.05). Mitochondrial membrane potential increased significantly in the point Neiguan electroacupuncture group compared with the model, point Huantiao electroacupuncture and sham operation groups (P<0.01,P<0.05). There was no statistically significant difference in mitochondrial membrane potential between the model and point Huantiao electroacupuncture groups (P>0.05). Conclusion Point Neiguan electroacupuncture pretreatment has a preventive protecting effect on MIRI rats. It produces a protecting effect on myocardium by increasing the NO content, strengthening NOS activity, reducing a decrease in cardiomyocyte mitochondrial membrane potential and inhibiting apoptosis.

Effect of Point Neiguan(PC6) Electroacupuncture Pretreatment on NO, NOS and Mitochondrial Membrane Potential in Rats with Myocardial Ischemia/Reperfusion Injury / 上海针灸杂志

Objective To explore the effect of point Neiguan(PC6) electroacupuncture pretreatment on nitric oxide (NO), nitric oxide synthase (NOS) and mitochondrial membrane potential by determining NO, NOS and mitochondrial membrane potential in rats with myocardial ischemia/reperfusion injury (MIRI).Method Forty male SD rats were randomized to sham operation, ischemia/reperfusion model, point Neiguan electroacupuncture and point Huantiao(GB30) electroacupuncture groups, 10 rats each. The model was made by coronary artery ligation. Before model making, electroacupuncture was given to the point Neiguan electroacupuncture and point Huantiao electroacupuncture groups, 20 min/d for a total of 7 d. T wave value in ECG leadⅡ was measured before and after model making. Myocardial pathomorphological changes were examined by HE staining. Serum NO and NOS contents were measured by a colorimetric nitrate reductase assay. Cardiomyocyte mitochondrial membrane potential was determined by fluorescence techniques.Result Serum NO and NOS contents and mitochondrial membrane potential decreased significantly in the model group compared with the sham operation group (P<0.05). Serum NO and NOS contents increased significantly in the point Neiguan electroacupuncture group compared with the model, sham operation and point Huantiao electroacupuncture groups (P<0.01,P<0.05). Mitochondrial membrane potential increased significantly in the point Neiguan electroacupuncture group compared with the model, point Huantiao electroacupuncture and sham operation groups (P<0.01,P<0.05). There was no statistically significant difference in mitochondrial membrane potential between the model and point Huantiao electroacupuncture groups (P>0.05). Conclusion Point Neiguan electroacupuncture pretreatment has a preventive protecting effect on MIRI rats. It produces a protecting effect on myocardium by increasing the NO content, strengthening NOS activity, reducing a decrease in cardiomyocyte mitochondrial membrane potential and inhibiting apoptosis.

Eukaryotic Expression and Immunogenic Research of Recombination Ebola Virus Membrane Protein Gp-Fc / 病毒学报

We used 293 cells to express the recombinant membrane protein of the Ebola virus. Then, the immunogenicity of the recombinant protein was studied by immunized BALB/c mice. According to the codon use frequency of humans, the gene encoding the extracellular domain of the Ebola virus membrane protein was optimized, synthesized, and inserted into the eukaryotic expression plasmid pXG-Fc to construct the human IgG Fc and Ebola GP fusion protein expression plasmid pXG-modGP-Fc. To achieve expression, the fusion protein expression vector was transfected into high-density 293 cells using transient transfection technology. The recombinant protein was purified by protein A affinity chromatography. BALB/c mice were immunized with the purified fusion protein, and serum antibody titers evaluated by an indirect enzyme-linked immunosorbent assay (ELISA). Purification and analyses of the protein revealed that the eukaryotic expression vector could express the recombinant protein GP-Fc effectively, and that the recombinant protein in the supernatant of the cell culture was present as a dimer. After immunization with the purified recombinant protein, a high titer of antigen-specific IgG could be detected in the serum of immunized mice by indirect ELISA, showing that the recombinant protein had good immunogenicity. These data suggest that we obtained a recombinant protein with good immunogenicity. Our study is the basis for development of a vaccine against the Ebola virus and for screening of monoclonal antibodies.

Effects of crocetin on VCAM-1 expression in human umbilical vein endothelial cells and monocyte-endothelial cell adhesion / 药学学报

Crocetin, a naturally occurring carotenoid, possesses antioxidant and antiatherosclerotic properties, of which the underlying mechanism remains unclear. In the present study, we examined the effects of crocetin (0.1, 1, 10 μmol·L(-1)) on angiotensin II (Ang II, 0.1 μmol·L(-1)) induced expression of vascular cell adhesion molecule-1 (VCAM-1) in human umbilical vein endothelial cells (HUVECs) and monocyte-endothelial cell adhesion. The effects of crocetin on the activation of nuclear factor kappa B (NF-κB) and intracellular reactive oxygen species (ROS) were also observed. The results demonstrated that crocetin notably suppressed Ang II induced NF-κB activation (P<0.01) and VCAM-1 expression (P<0.05, P<0.01) in HUVECs, accompanied by a markedly reduced monocyte-endothelial cell adhesion (P<0.05, P<0.01). In addition, preincubation with crocetin resulted in a significant enhancement of cellular antioxidant capacity (P<0.05, P<0.01), while Ang II induced intracellular ROS decreased markedly (P<0.05, P<0.01). These results indicated that crocetin was capable of suppressing Ang II induced VCAM-1 expression and monocyte-endothelial cell adhesion by suppression of NF-κB activation, which might be derived from the enhancement of antioxidant capacity and subsequent reduction of intracellular ROS.

Effect of sesamin on pulmonary vascular remodeling in rats with monocrotaline-induced pulmonary hypertension / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of sesamin (Ses) on pulmonary vascular remodeling in rats with monocrotaline ( MCT)-induced pulmonary hypertension (PH).</p><p><b>METHOD</b>Totally 48 male Sprague-Dawley (SD) rats were fed adaptively for one week and then divided into the normal control group, the MCT group, the MCT +Ses (50 mg x kg(-1)) group and the MCT + Ses (100 mg x kg(-1)) group, with 12 rats in each group. The PH rat model was induced through the subcutaneous injection with MCT(60 mg x kg(-1)). After the administration for four weeks, efforts were made to measure the right ventricular systolic pressure( RVSP) and mean pulmonary artery pressure (mPAP) through right jugular vein catheterization, and isolate right ventricle( RV) and left ventricle( LV) +septum (S) and measure their length to calculate RV/ ( LV + S) and ratio of RV to tibial length. Pathologic changes in arterioles were observed by HE staining. Masson's trichrome stain was used to demonstrate changes in collagen deposition of arterioles. The alpha-smooth muscle actin (alpha-SMA) expression in pulmonary arteries was measured by immunohistochemisty. The total antioxidative capacity (T-AOC) and malondialdehyde (MDA) content in pulmonary arteries were determined by the colorimetric method. The protein expressions of collagen I, NOX2 and NOX4 were analyzed by Real-time PCR and Western blot.</p><p><b>RESULT</b>After the administration for 4 weeks, Ses could attenuate RVSP and mPAP induced by MCT, RV/ (LV + S) and ratio of RV to Tibial length, alpha-SMA and collagen I expressions and remodeling of pulmonary vessels and right ventricle. Meanwhile, Ses could obviously inhibit the expressions of NOX2, NOX4 and MDA content and increase T-AOC.</p><p><b>CONCLUSION</b>Sesamin could ameliorate pulmonary vascular remodeling induced by monocrotaline in PH rats. Its mechanism may be related to expressions of NOX2 and NOX4 expression and reduction in oxidative stress injury.</p>

Effect of chrysin on expression of NOX4 and NF-κB in right ventricle of monocrotaline-induced pulmonary arterial hypertension of rats / 药学学报

The aim of the present study is to investigate the protective effect of chrysin (5,7-dihydroxyflavone) on right ventricular remodeling in a rat model of monocrotaline-induced pulmonary arterial hypertension (PAH). PAH rats were induced by a single injection of monocrotaline (60 mg x kg(-1), sc) and were administered with chrysin (50 or 100 mg x kg(-1) x d(-1)) for 4 weeks. At the end of experiment, the right ventricular systolic pressure (RVSP) and mean pulmonary artery pressure (mPAP) were monitored via the right jugular vein catheterization into the right ventricle. Right ventricle (RV) to left ventricle (LV) + septum (S) and RV to tibial length were calculated. Right ventricular morphological change was observed by HE staining. Masson's trichrome stain was used to demonstrate collagen deposition. The total antioxidative capacity (T-AOC) and malondialdehyde (MDA) levels in right ventricle were determined according to the manufacturer's instructions. The expressions of collagen I, collagen III, NADPH oxidase 4 (NOX4) and nuclear factor-kappa B (NF-κB) were analyzed by immunohistochemisty, qPCR and (or) Western blot. The results showed that chrysin treatment for 4 weeks attenuated RVSP, mPAP and right ventricular remodeling index (RV/LV+S and RV/Tibial length) of PAH rats induced by monocrotaline. Furthermore, monocrotaline-induced right ventricular collagen accumulation and collagen I and collagen III expression were both significantly suppressed by chrysin. The expressions of NOX4, NF-κB and MDA contents were obviously decreased, while the T-AOC was significantly increased in right ventricule from PAH rats with chrysin treatment. These results suggest that chrysin ameliorates right ventricular remodeling of PAH induced by monocrotaline in rats through its down-regulating of NOX4 expression and antioxidant activity, and inhibiting NF-κB expression and collagen accumulation.

Effect of sequoyitol on expression of NOX4 and eNOS induced with glucose in human umbilical vein endothelial cells / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the protective effect and mechanism of sequoyitol (Sep) on high glucose-induced human umbilical vein endothelial cells (HUVECs) injury.</p><p><b>METHODS</b>HUVECs were cultured with high glucose (30 mmol/L) in the presence or absence of sequoyitol (0.1, 1 and 10 micromol/L) for 24 h. Cell proliferation was measured by BrdU marking and cell cycle was detected by flow cytometry. 2', 7'-dichlorofluorescein diacetate was used to evaluate intracellular reactive oxygen species (ROS) levels. The NO, malonydialdehyde (MDA) and H2O2 levels were determined by colorimetric method according to the manufacturer's instructions. The expression of endothelial nitric oxide synthase (eNOS) and NADPH oxidase 4 (NOX4) were measured by real-time PCR and Western blot.</p><p><b>RESULTS</b>In the present study, we found that sequoyitol pretreatment for 1 h significantly decreased cell injury, promoted cell proliferation. Meanwhile sequoyitol significantly down-regulated NOX4 expression and decreased the level of ROS, MDA and H2O2 and obviously increased NO levels and up-regulated eNOS expression.</p><p><b>CONCLUSION</b>Sequoyitol alleviates high glucose-induced cell injuries in HUVECs via inhibiting oxidative stress and up-regulating eNOS expression.</p>

Effect of sequoyitol on expression of NOX4 and eNOS in aortas of type 2 diabetic rats / 药学学报

The aim of the present study is to investigate the effects of sequoyitol (Seq) on expression of eNOS and NOX4 in aortas of type 2 diabetic rats. Type 2 diabetic rats induced by high fat and high sugar diet and low dose of streptozotocin (STZ, 35 mg x kg(-1)) and were administered Seq (12.5, 25 and 50 mg x kg(-1) x d(-1)) for 6 weeks. The fasting blood glucose (FBG) and body weight were tested. Acetylcholine (Ach) induced endothelium-dependent relaxation and sodium nitroprusside (SNP) induced endothelium-independent relaxation were measured in aortas for estimating endothelial function. Aortic morphological change was observed with HE staining. The level of serum insulin was measured by radioimmunoassay. The total antioxidative capacity (T-AOC), malondialdehyde (MDA) and NO levels in aortas were determined according to the manufacturer's instructions. In addition, the expressions of eNOS and NOX4 in aortas were measured by immunohistochemisty, real-time PCR or Western blotting. The results showed that Seq significantly decreased FBG and insulin resistance, and improved aortic endothelium-dependent vasorelaxation function. The expressions of NOX4 and MDA content were obviously decreased, while the expression of eNOS, the levels of NO and T-AOC increased significantly in aortas of diabetic rats with Seq treatment. In conclusion, Seq protects against aortic endothelial dysfunction of type 2 diabetic rats through down-regulating expression of NOX4 and up-regulating eNOS expression.

CT and MRI findings in patients with autoimmune pancreatitis / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate computed tomography (CT) and magnetic resonance imaging (MRI) findings in patients with autoimmune pancreatitis (AIP).</p><p><b>METHODS</b>The imaging findings of pancreas and extra-pancreas in 24 patients with AIP were retrospectively reviewed. Among them, CT scan was performed in 18 patients, MRI in 11, and bGth CT and MRI in 10.</p><p><b>RESULTS</b>The pancreas showed diffuse enlargement (25%, 6/24), focal enlargement (37. 5%, 9/24), combined enlargement (25%, 6/24) ,and no enlargement (12. 5%, 9/24). Unenhanced CT showed hypoattenuation in AIP area (n = 2) . After intravenous injection of contrast medium, 17 patients showed abnormal contrast enhancement in the affected pancreatic parenchyma, including hypoattenuation during the arterial phase (50%, 9/18) and hyper attenuation during the delayed phase (94. 4%, 17/18). Precontrast MRI showed abnormal signal intense (n =9), including hypointense on T1-weight images (T1 WI) (n = 7), hyperintense (n = 7) and hypointense (n = 2) on T2-weight images (TIWI). Enhanced MRI demonstrated abnormal contrast enhancement within lesions (n = 11), including hypoattenuation during the arterial phase (81. 8%, 9/11) and good enhancement during the delayed phase (100%, 11111). A capsule-like rim was seen around pancreas (37. 5%, 9/24), among which CT detected in 6 out of 18 patients and MRI found in 7 out of 11 patients.The main pancreatic duct lumen within lesions has no visualization (100%, 24/24) and upstream dilation of the main pancreatic duct (n = 8) , ranging from 2. 2 to 4. 5 mm(mean 3. 1 0. 47 mm) in diameter. Narrowing of the common bile duct was shown in 14 patients. Miscellaneous findings were: infiltration of extrapancreatic vein (n = 9) and artery (n = 1); mild fluid collection around pancreas (n = 2); pseudocysts (n = 3). Fourteen patients also presented one or more of the following extrapancreatic imaging findings: narrowing of the intra-hepatic bile duct or hilar duct (n = 5); thickening of gallbladder wall (n = 5); fibrosis in mesenteric (n = 2), in retroperitoneal (n = 2) and in ligamentum teres hepatis (n = 1); renal involvement (n = 3); peri-pancreatic or para-aortic lymphadenopathy (n = 10); and ulcerative colitis (n = 3).</p><p><b>CONCLUSION</b>AIP display some characteristic CT and MRI imaging features: sausage-like change of the pancreas; capsule-like rims around lesions; delayed contrast enhancement in the affected pancreatic parenchyma; segment or diffuse pancreatic duct stenosis but mild upstream dilation and extrapancreatic organs involvement. CT and MRI findings combining with serological tests and pancreas biopsy can assist physicians to make accurate and timely diagnosis.</p>

Effects of rutaecarpine on right ventriclar remodeling in rats with monocrotaline-induced pulmonary hypertension / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the protective effects of rutaecarpine (Rut) on right ventricular remodeling in rats with monocrotaline-induced pulmonary hypertension (PH).</p><p><b>METHOD</b>Forty-eight SD rats were fed adaptively for 1 week and then were randomly divided into the following 4 groups (n = 12): normal control group, monocrotaline (MCT) treatment group, MCT treatment with Rut (20 mg/kg)group and MCT treatment with Rut (40 mg/kg) group. PH rats were induced by a single injection of monocrotaline (60 mg/kg, sc) and were administered with Rut (20 or 40 mg/kg/d) for 4 weeks. At the end of experiment, the right ventricular systolic pressure (RVSP) and mean pulmonary artery pressure (mPAP) were monitored via the right jugular vein catheterization into the right ventricle. The ratio of right ventricle (RV) to left ventricle (LV) + septum (S) and the ratio of RV to tibial length were calculated. Right ventricular morphological changes were deserved by HE staining. Masson's trichrome staining was used to display collagen deposition. The total antioxidative capacity (T-AOC) and malondialdehyde (MDA) levels in right ventricle were determined according to the manufacturer's instructions. mRNA and protein expression levels of NOX4, collagen I and collagen III were analyzed by immunohistochemisty, real-time PCR and Western blot.</p><p><b>RESULTS</b>The results showed that Rut treatment for 4 weeks attenuated RVSP, mPAP and right ventricular remodeling index (RV/LV + S and RV/Tibial length) of PH rats induced by monocrotaline. Furthermore, the right ventricular collagen deposition and collagen I and collagen I expression induced by MCT were both significantly suppressed by Rut. The expression levels of NOX4 and MDA were obviously decreased, while the T-AOC was significantly increased in right ventricular from PH rats treated with Rut.</p><p><b>CONCLUSION</b>These results suggested that Rut ameliorates the right ventricular remodeling in rats with PH induced by MCT through down-regulating of NOX4 expression and collagen accumulation.</p>

Effects of sequoyitol on expression of NADPH oxidase subunits p22 phox and p47 phox in rats with type 2 diabetic liver disease / 药学学报

This study is to observe the effects of sequoyitol on the expression of NADPH oxidase subunits p22 phox and p47 phox in rats with type 2 diabetic liver diseases. The model of high fat and high sugar diet as well as intraperitoneal injection of small dose of streptozotocin (STZ, 35 mg x kg(-1)) induced diabetic rat liver disease was used. After sequoyitol (50, 25 and 12.5 mg x kg(-1)) was administrated for 6 weeks, the contents of blood glucose (BG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total antioxidant capacity (T-AOC), hydrogen peroxide (H2O2), NO and insulin (Ins) were measured, liver p22 phox and p47 phox mRNA content was determined with real-time PCR and the expression of p22 phox and p47 phox protein was examined by Western blotting. In addition, pathological changes in liver were observed with HE staining. Sequoyitol could reduce the content of fasting blood glucose, ALT, AST, Ins and H2O2, restore insulin sensitive index (ISI) and weight, elevate liver tissue T-AOC and NO content, reduce the NADPH oxidase subunit liver tissue p22 phox and p47 phox mRNA and protein expression, as well as ameliorate liver pathologic lesions. The results showed that sequoyitol can ease the type 2 diabetic rat liver oxidative stress by lowering NADPH oxidase expression.