Objective:To explore the clinical significance of interleukin 6(IL 6) in pathogenesis of diabetic nephropathy by comparing the plasma and urinary level of IL 6 in patients with type 2 diabetes mellitus(DM).Methods:The plasma and urinary level of IL 6 were measured with radioimmunoassay in 20 normal controls(NC group) and 45 cases of type 2 DM,including 23 cases without albuminuria(DSM group) and 22 cases with trace albuminuria(EDN group).Results:(1)The plasms and urinary level of IL 6 markly elevated in SDM group(P

Objective To develop a sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of forsklin in rat plasma.Methods After extraction with methyl tert-butyl ether,chromatographic separation was performed on a C18 column with the mobile phase consisting of water ( 0.1％ formic acid)-acetonitrile in a gradient elution mode.A tandem mass spectrometer equipped with electrospray ionization (ESI) source was used as detector in the positive ion mode.Quantification was performed using multiple reaction monitoring (MRM) with the precursor product combination ions of m/z 411→375.3 and 285→193 for forsklin and diazepam.Results Good linearity was obtained in the 0.5-1000 ng/ml range for the analyte and the analytical method was validated in terms of specificity,precision,accuracy,recovery,stability and matrix effect.These assays gave RSD values always lower than 14.4％ and RE values between -3.5 ％ and 3.8％.In addition,the specificity,extraction recovery,stability and matrix effect were satisfactory.Conclusion Due to its high sensitivity,specificity and simplicity,the method could be used for pharmacokinetic studies of forsklin.

A method was developed for the determination of zeranols (α,β-zearalanol,α,β-zearalenol,zear-alanone,zearalenone) with RRLC-MS/MS in the plant tissue. The samples were extracted with acetonitrile and reextracted with aqueous alkaline solution,cleaned up with MAX column and then determined by RRLC-MS/MS using multiple reaction monitoring ( MRM) scan mode. The results showed that the working curves were linear in the range of 0 -20 μg/kg. The limits of detection ( LOD) of zeranols were 0.5 μg/kg and the limits of quantification (LOQ) was 1. 0μg/kg. Extraction recoveries for zeranols ranged from 75. 8% to 105.4% and RSD was between 2.4% and 12.1%. The method is suitable for the determination of zeranols in the plant tissue.

To facilitate noninvasive diagnosis of anemia, high-performance and portable near infrared (NIR) spectrometer for human blood constituents was designed and fabricated based on linear variable filter (LVF).Meanwhile, the performance of support vector regression (SVR) model for quantitative analysis of human hemoglobin (Hb) was investigated.Spectral data were collected noninvasively from 100 volunteers by self-designed LVF-NIR spectrometer, then divided into calibration set, validation set 1 and 2.To establish a robust SVR model, grid search method was applied to optimize the penalty parameter and kernel function parameter c=5.28, g=0.33.Then, Hb levels in the validation 1 and 2 sets were quantitatively analyzed.The results showed that the root mean square error of prediction (RMSEP) were 10.20 g/L and 10.85 g/L, respectively, and the relative RMSEP (R-RMSEP) were 6.85% and 7.48%, respectively.The results indicated that the SVR model had high prediction accuracy to Hb level and adaptability to different samples, and could satisfy the requirements of clinical measurement.Based on the SVR algorithm, the self-designed LVF-NIR spectrometer has a wide application prospect in the field of non-invasive anemia diagnosis.

Objective To explore the clinical effect of magnetic resonance imaging (MRI) and CT scanning in the diagnosis of acute craniocerebral injury.Methods From October 2012 to November 2016,100 patients with acute craniocerebral injury in Zhoushan Hospital were selected.According to the order of patients admitted to hospital,they were randomly divided into observation group and control group,with 50 cases in each group.The observation group received MRI inspection,the control group received CT scanning.The clinical examination results of the two groups were compared.Results The positive rate and missed diagnosis rate of the observation group were 98.0％,2.0％,respectively,which of the control group were 72.0％,24.0％,respectively,the differences were statistically significant (x2 =26.958,17.468,all P ＜ 0.05).The detection rate of craniocerebral injury position lesion in the observation group was significantly higher than control group,the differences between the two groups was statistically significant (x2 =5.030,5.110,11.250,5.430,all P ＜ 0.05).The clinical total effective rate of the observation group was 94.0％,which was higher than 78.0％ of the control group,the difference was statistically significant (x2 =15.432,P ＜ 0.05).Conclusion The clinical value of MRI in the diagnosis of acute craniocerebral injury issuperior to CT scanning.

Objective: To evaluate the clinical value of CT combined with MRI in the qualitative diagnosis of hepatic lesions. Methods: From August 2015 to March 2018, 100 patients with liver diseases were selected from Zhoushan Hospital.CT, MRI and combined examination were performed respectively.The results of diagnosis of liver diseases in three different ways were compared and analyzed. Results: A total of 96 lesions were detected by CT, the detectable rate was 88.89%, 97 lesions were detected by MRI, and the detectable rate was 89.81%.A total of 105 lesions were detected by CT and MRI, and the detectable rate was 97.22%.The detection rate and number of detected lesions of combined test were significantly higher than those of single test (χ²=8.25, 6.22, all P<0.05), but there were no statistically significant differences in the detection rate and the number of lesions between CT and MRI (all P>0.05). Conclusion The diagnosis of benign and malignant lesions by the combination of CT and MRI is very good.The diagnostic effect is relatively small and the safety factor is high.It is worth widely used in clinical diagnosis of patients.

Subolesin (4D8), the ortholog of insect akirins, is a highly conserved protective antigen and thus has the potential for development of a broad-spectrum vaccine against ticks and mosquitoes. To date, no protective antigens have been characterized nor tested as candidate vaccines against Dermacentor silvarum bites and transmission of associated pathogens. In this study, we cloned the open reading frame (ORF) of D. silvarum 4D8 cDNA (Ds4D8), which consisted of 498 bp encoding 165 amino acid residues. The results of sequence alignments and phylogenetic analysis demonstrated that D. silvarum 4D8 (Ds4D8) is highly conserved showing more than 81% identity of amino acid sequences with those of other hard ticks. Additionally, Ds4D8 containing restriction sites was ligated into the pET-32(a+) expression vector and the recombinant plasmid was transformed into Escherichia coli rosetta. The recombinant Ds4D8 (rDs4D8) was induced by isopropyl beta-D-thiogalactopyranoside (IPTG) and purified using Ni affinity chromatography. The SDS-PAGE results showed that the molecular weight of rDs4D8 was 40 kDa, which was consistent with the expected molecular mass considering 22 kDa histidine-tagged thioredoxin (TRX) protein from the expression vector. Western blot results showed that rabbit anti-D. silvarum serum recognized the expressed rDs4D8, suggesting an immune response against rDs4D8. These results provided the basis for developing a candidate vaccine against D. silvarum ticks and transmission of associated pathogens.
Animals ; Antigens/chemistry/genetics/*immunology/isolation & purification ; Arthropod Proteins/chemistry/genetics/*immunology/isolation & purification ; Chromatography, Affinity ; Cloning, Molecular ; Cluster Analysis ; Conserved Sequence ; Dermacentor/*genetics ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli/genetics ; Gene Expression ; Humans ; Molecular Sequence Data ; Molecular Weight ; Phylogeny ; Recombinant Proteins/chemistry/genetics/immunology/isolation & purification ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid

Vaccination is considered a promising alternative for controlling tick infestations. Haemaphysalis longicornis midgut proteins separated by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membrane were screened for protective value against bites. The western blot demonstrated the immunogenicity of 92 kDa protein (P92). The analysis of the P92 amino acid sequence by LC-MS/MS indicated that it was a H. longicornis paramyosin (Hl-Pmy). The full lenghth cDNA of Hl-Pmy was obtained by rapid amplification of cDNA ends (RACE) which consisted of 2,783 bp with a 161 bp 3' untranslated region. Sequence alignment of tick paramyosin (Pmy) showed that Hl-Pmy shared a high level of conservation among ticks. Comparison with the protective epitope sequence of other invertebrate Pmy, it was calculated that the protective epitope of Hl-Pmy was a peptide (LEEAEGSSETVVEMNKKRDTE) named LEE, which was close to the N-terminal of Hl-Pmy protein. The secondary structure analysis suggested that LEE had non-helical segments within an alpha-helical structure. These results provide the basis for developing a vaccine against biting H. longicornis ticks.
Amino Acid Sequence ; Animals ; Antigens/genetics/*metabolism ; Base Sequence ; Blotting, Western ; Chromatography, Liquid ; *Cloning, Molecular ; DNA, Complementary/genetics ; Epitopes ; Gene Expression Regulation/*physiology ; Ixodidae/genetics/*metabolism ; Molecular Sequence Data ; Tandem Mass Spectrometry

In order to explore tick proteins as potential targets for further developing vaccine against ticks, the total proteins of unfed female Dermacentor silvarum were screened with anti-D. silvarum serum produced from rabbits. The results of western blot showed that 3 antigenic proteins of about 100, 68, and 52 kDa were detected by polyclonal antibodies, which means that they probably have immunogenicity. Then, unfed female tick proteins were separated by 12% SDS-PAGE, and target proteins (100, 68, and 52 kDa) were cut and analyzed by LC-MS/MS, respectively. The comparative results of peptide sequences showed that they might be vitellogenin (Vg), heat shock protein 60 (Hsp60), and fructose-1, 6-bisphosphate aldolase (FBA), respectively. These data will lay the foundation for the further validation of antigenic proteins to prevent infestation and diseases transmitted by D. silvarum.
Animals ; Antigens/*chemistry/immunology ; Arthropod Proteins/*chemistry/immunology ; Electrophoresis, Polyacrylamide Gel ; Female ; Ixodidae/*chemistry/immunology ; Molecular Weight ; Rabbits ; Tandem Mass Spectrometry

Caenorhabditis elegans hid-1 gene was first identified in a screen for mutants with a high-temperature-induced dauer formation (Hid) phenotype. Despite the fact that the hid-1 gene encodes a novel protein (HID-1) which is highly conserved from Caenorhabditis elegans to mammals, the domain structure, subcellular localization, and exact function of HID-1 remain unknown. Previous studies and various bioinformatic softwares predicted that HID-1 contained many transmembrane domains but no known functional domain. In this study, we revealed that mammalian HID-1 localized to the medial- and trans- Golgi apparatus as well as the cytosol, and the localization was sensitive to brefeldin A treatment. Next, we demonstrated that HID-1 was a peripheral membrane protein and dynamically shuttled between the Golgi apparatus and the cytosol. Finally, we verified that a conserved N-terminal myristoylation site was required for HID-1 binding to the Golgi apparatus. We propose that HID-1 is probably involved in the intracellular trafficking within the Golgi region.
Animals ; Brefeldin A ; pharmacology ; Cell Line, Tumor ; Cytosol ; drug effects ; metabolism ; Humans ; Intracellular Space ; drug effects ; metabolism ; Membrane Proteins ; metabolism ; Protein Transport ; drug effects ; Rats ; Vesicular Transport Proteins ; metabolism ; trans-Golgi Network ; drug effects ; metabolism