AIM: To establish variation in flow rates of anthraquinones in Anzhong Tongbian Capsule(Radix et Rhizoma rhei,Radix paeoniae alba,Fructus cannabis and Semen trichosanthis). METHODS: The thin layer chromatographic method was adopted. The anthraquinones compounds from Radix et Rhizoma Rhei in Anzhong Tongbian Capsules were extracted by ultrasound and separated by outspread on silica gel G thin layer,and n-hexane-ethyl acetate-formic acid(30 ∶ 10 ∶ 0. 5)as the developing system,detection wavelength was set at 435 nm and reference wavelength was 610 nm by using CAMAG TLC ScannerⅢ. RESULTS: Anthraquinones was separated well and 5 peaks of emodin,chrysophanol,aloe-emodin,rhein and physcion were ascertained. CONCLUSION: The method is simple and accurate with a good reproducibility and can be used for the quality control for Radix et Rhizoma Rhei in Anzhong Tongbian Capsules.

Objective To evaluate the serum 25-hydroxy vitamin D 3 level in newly diagnosed type 2 diabe-tes,and its relationship with function of islet beta cell .Methods 60 newly diagnosed type 2 diabetes patients and 48 normal glucose tolerance healthy control cases were selected .The level of serum 25-hydroxy vitamin D3 of two groups were measured using the way of high performance liquid chromatograph .The difference was compared between two groups.Insulin resistance index(HOMA-IR) and islet beta cell secrete index(HOMA-β) was used to respectively es-timate insulin sensitivity and islet beta cell function in type 2 diabetes group .The correlation was analyzed between 25-hydroxy vitamin D 3 and fasting blood glucose ,HOMA-IR,HOMA-β.Results The level of serum 25-hydroxy vita-min D3 (28.68 ±1.61) ng/mL in type 2 diabetes group was significantly lower than that of control group (41.30 ± 1.12)ng/mL(t=3.47,P<0.01).There is negative correlation between 25-hydroxyl vitamin D3 and fasting glucose (r=0.48,P<0.05).There was positive correlation between 25-hydroxy vitamin D3 and HOMA-IR(r=0.52,P<0.05).There was no correlation between 25-hydroxy vitamin D3 and HOMA-β(r=-0.06,P>0.05).Conclusion The level of 25-hydroxy vitamin D 3 was significantly low in newly diagnosed type 2 diabetes patients .Low serum 25-hydroxyl vitamin D 3 level affects islet beta secretion cell function ,the supplementation of vitamin D 3 may be a simple and effective method to reduce the ocurrence of type 2 diabetes .

Objective To establish an HPLC method for the content determination of sodium Danshensu inBawei Qidan Capsule.Methods HPLC was applied and the determination was performed on Shim-pack CLC-ODS C18 column (250 mm×4.6 mm, 5μm) with methanol-water- acetic acid (7∶92∶1, V/V/V) solution at a flow rate of 1.0 mL/min and detection wavelength at 281 nm. The column temperature was 30℃, and injection volume was 10μL.Results Through methodological study, the linear range of sodium Danshensu was 0.494 4-4.944μg (r=0.9996), the average recovery was 97.20%, RSD=1.13%.Conclusion The HPLC method for the content determination of sodium Danshensu inBawei QidanCapsule was user-friendly, accurate and reliable, with good repeatability and stability, which can be used for quality control ofBawei Qidan Capsule.

AIM:Atherosclerosis primarily involved systemic arteries .Luminal surface , a monolayer of endothelial cells , of artery directly exposes to blood and is susceptible to active substances in the blood .Exosomes contain significantly amount of proteins and RNAs .Ex-osomes can be good and bad for cells , depending on their component .Thus, exosomes may contribute to atherosclerosis by affecting endothelial cells .This study analyzed the relationship of exosome proteins and atherosclerosis .METHODS: Fifty-six patients and healthy subjects were recruited and divided into two comparisons:healthy subjects vs atherosclerosis ( HS vs AS) , and hypertension vs hypertension plus atherosclerosis ( HT vs HT+AS) .Serum exosomes were decoded by protein mass spectrometry .The protein profile and function were analyzed by gene ontology ( GO) .RESULTS:It was found that five child terms repeatedly appeared in “response to stimulus” and “immune system process” of BP of the two categories ( HS vs AS and AS vs HT+AS):“positive regulation of innate immune response”,“immune response-activating signal transduction”,”activation of innate immune response”,“innate immune re-sponse-activating signal transduction” and “innate immune response activating cell surface receptor signaling pathway ”.Two child terms repeatedly showed in “binding” of MF of the two categories:“antigen binding” and “enzyme binding”.Two proteins, PSMA6 and PSMA7, were repeatedly shown in the two categories .CONCLUSION:GO analysis was utilized for structure hierarchy “tree” to illustrate these proteins involved in various terms in BP , CC and MF.The PPI analysis supplied proteins which may play potentially im-portant roles in AS process .Innate immune system and blood coagulation pathway contribute to AS formation .The proteins, PSMA6, PSMA7 and Annexin A2, may can be the new target proteins for prevention and treatment of AS .

AIM:To analyze the proteins included in exosomes derived from blood of patients with hypertension and seek the main pathologi -cal changes in hypertension .METHODS:Forty-seven patients and healthy subjects were recruited and divided into two comparisons :healthy subjects vs atherosclerosis ( HS vs AS) , and atherosclerosis vs hypertension plus atherosclerosis ( AS vs HT+AS) .We extrac-ted exosomes from blood and utilized LC-MS/MS to identify the protein expression .We used GO analysis to established the hierarchy programs of biological process and molecular function .PPI was used to find the proteins related to the terms .RESULTS:It was found that three final child terms repeatedly shown in BP of the two categories ( HS vs AS and AS vs HT+AS):“signal transduction in re-sponse to DNA damage”,“response to zinc ion”, and“platelet aggregation”.It was found that two final child terms in MF of the two categories:“interleukin 2 receptor binding” and“ploy(A) RNA binding”.The proteins, PSMA6, PSMA7 and CA2, were related to the terms in the two categories .CONCLUSION: We discovered that the exosome proteins may indicate the pathological changes in hypertension through the biological processes related with the specific proteins .These specific proteins, such as VCL, PSMA6, DP, AKAP, ATP5B and CA2, can be the new indicators for severity of hypertension and new therapeutic targets .

Objective:To express and purify the TRIM5? chimaera[TRIM5? H(R328-332)] protein and to explore the interaction between the TRIM5? H(R328-332)and HIV-1gag. Methods:The plasmid pET28aTRIM5? H(R328-332) was transformed to E.coli BL21 (DE3) strain ,and the expression of TRIM5? H(R328-332) protein was induced by IPTG,purified with Ni2+ chromatography.The expression and purification of TRIM5? H(R328-332) were analyzed by SDS-PAGE and Western blot,and the interaction between TRIM5? H(R328-332) and HIV-1gag was detected by co-immunoprecipitation,His pull-down and ELISA. Results:The recombinant plasmid pET28aTRIM5? H(R328-332) was successfully expressed in E.coli. The results showed that the purified full length TRIM5? H(R328-332) interacted with HIV-1gag protein. Conclusion:The human TRIM5? chimaera was expressed successfully in vitro,and the study demonstrates that the human TRIM5? chimaera interacts with HIV-1 gag in vitro.

Objective To analyze the dielectrophoresis patterns of the proteome of the Rifampin-dependent and-resistant stains of Mycobacterium tuberculosis,to search and identify the differently expressed proteins,and to provide the proteomic basis for researching the mechanism of anti-tuberculosis drug dependence of M.tuberculosis.Methods The whole somatic proteins were extracted from two strains of M.tuberculosis.The first dimensional ampholine electrophoresis was performed on immobilized pH gradient(IPG) rod gels(pH 4-7).Then the proteins on IPG strips were separated using SDS-PAGE.The stained gels were scanned with image scanner and the images were analyzed by Imagemaster 2D software.The differentially expressed proteins were detected by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS).Results Seven hundred and fifty-three spots were detected in Rifampin-dependent strain of M.tuberculosis,while 584 spots were detected in Rifampin-resistant strain,including 404 match spots(the match rate: 61.5%).As to the expression in Rifampin-dependent strain,7 spots significantly up-regulated and 35 spots down-regulated,6 spots were absent in expression,and 5 spots expressed separately,most of the spots were small molecular proteins.Ten spots were selected to run MS analysis.Nine spots were identified as representing 7 proteins.Conclusion The Rifampin-dependent strain of M.tuberculosis is characterized by a rapid and vigorous growth mainly by means of the differential expression of enzymes related to energy metabolism and fatty acid biosynthesis.

In order to investigate the prevalence and track genetic and antigenic evolutions of infectious bronchitis virus (IBV) and their prevalence in Guangxi, China since 1985, gene amplification and sequencing and virus neutralization (VN) test on chicken embryo tracheal organ cultures were used in genotyping and serotyping of 28 IBV isolates during 2009-2011 in Guangxi. The results of N gene sequencing and comparison showed that the 28 isolates and reference strains were classified into three groups, and most isolates belonged to group Ill, while the isolates in 1985-2008 belonged to groups IV and II. The data of VN test indicated that the 28 isolates belonged to 6 serotypes; among them, 71. 4% belonged to serotypes 1, 2, and 3, and 11 (39.3%) shared the same serotype with the current vaccine strains. Given the data of our previous study, it is found that prevalent serotypes and their proportions varied in different areas of Guangxi and during different periods. These data lay a good foundation for developing an oil-emulsified inactivated polyvalent vaccine containing local dominant serotypes for the effective prevention and control of infectious bronchitis.
Animals ; Antibodies, Viral ; immunology ; Chick Embryo ; Chickens ; China ; epidemiology ; Coronavirus Infections ; epidemiology ; immunology ; veterinary ; virology ; Infectious bronchitis virus ; classification ; genetics ; immunology ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; epidemiology ; immunology ; virology

OBJECTIVETo screen biomarkers which can be used as an auxiliary method in the diagnosis of Henoch-Schönlein purpura (HSP) and to evaluate their diagnostic values by receiver operating characteristic (ROC) curve analysis.

METHODSA total of 127 children diagnosed with HSP between April 2012 and March 2014 were included in the HSP group and an equal number of healthy children were included in the control group. Twelve parameters, i.e., serum amyloid protein A (SAA), interleukin-6 (IL-6), immunoglobulins (IgA, IgG, IgM, and IgE), C-reactive protein (CRP), white blood cell (WBC) count, complements C3 and C4, anti-streptolysin O, and ferritin, were analyzed. The values of the screened biomarkers for diagnosis of HSP were assessed by ROC curve analysis.

RESULTSThe HSP group had significantly higher levels of SAA, IL-6, CRP, WBC, IgA, and IgM than the control group (P<0.05). The areas under the ROC curve of SAA, IL-6, WBC, IgA, and IgM for the diagnosis of HSP were higher than 0.7 (P<0.05). The optimal cut-off values of SAA, IgA, IgM, WBC, and IL-6 for the diagnosis of HSP were 3.035 μg/mL, 1579.5 mg/L, 922.5 mg/L, 8.850 × 10⁹/L, and 7.035 pg/mL, respectively; the corresponding sensitivities of the optimal cut-off values for the diagnosis of HSP were 95.1%, 75.6%, 72.3%, 78.0%, and 63.4%, respectively, and the corresponding specificities were 90.2%, 85.4%, 82.4%, 70.7%, and 80.5%, respectively.

CONCLUSIONSSAA, IgA, IgM, WBC, and IL-6 are valuable biomarkers for clinical diagnosis of HSP and among them SAA seems to be the best one.

Adolescent ; Biomarkers ; blood ; C-Reactive Protein ; analysis ; Child ; Child, Preschool ; Female ; Humans ; Male ; Purpura, Schoenlein-Henoch ; blood ; diagnosis ; ROC Curve ; Serum Amyloid A Protein ; analysis

Aim To investigate the regulatory effect of Cortaetin on pathological myocardial hypertrophy induced by isoprenaline (ISO) and the underlying mechanism. Methods ISO was used to stimulate neonatal rat cardiomyocytes for 24 h, and myocardial hypertrophy model was established at the cellular level. C57BL/6 mice were injected subcutaneously with ISO for one week to establish myocardial hypertrophy model at animal level. RT-qPCR was used to detect the changes of mRNA and Western blot was used to detect the changes of relative protein content. Immunofluorescence was used to measure the subcellular location of Cortaetin and the change of its expression. The overex-pression of Cortaetin by adenovirus infection and the knockdown of Cortaetin by transfection of small interfering RNA were studied. Results On the cellular and animal levels, ISO-induced myocardial hypertrophy models were successfully established, and it was observed that ISO caused the decrease of Cortaetin and N-cadherin protein levels. Overexpression of Cortaetin could reverse the decrease of N-cadherin protein level and myocardial hypertrophy caused by ISO. Knockdown of Cortaetin showed the opposite effect. Conclusion Cortaetin, in combination with N-cadherin, may play a role in combating myocardial hypertrophy by enhancing the connections between cardiomyocytes.