Objectives: To evaluate the immunogenicity of Methods: Protein extracts from Results: Immunization with Conclusion This is the advanced study to investigate the immunogenicity of
Animals ; Antibodies, Bacterial/immunology* ; Antigens, Bacterial/immunology* ; Bacterial Proteins/immunology* ; Cross Reactions ; Cytokines/immunology* ; Female ; Genome, Bacterial ; Immunoglobulin G/immunology* ; Immunoglobulin M/immunology* ; Macrophages/immunology* ; Mice, Inbred BALB C ; Mycobacterium avium Complex/immunology* ; Mycobacterium tuberculosis/immunology* ; Tuberculosis Vaccines/administration & dosage* ; Whole Genome Sequencing

OBJECTIVE To investigate the effect of neuroprotective effect of lychee seed saponins (LSS) in BV-2. METHODS Aβ1-42 induced BV-2 cells were incubated with LSS for 12 h, the content of the inflammatory factors such as IL-1β, TNF-α, COX-2 and iNOS in the supernatant of BV-2 cell were measured by ELISA. The detection of the mRNA levels and the protein expression of the inflammatory factors including IL-1β, TNF-α, COX-2 and iNOS using real-time PCR and Western blotting, respectively. RESULTS The level of IL-1β, COX-2 and iNOS significantly increased with the treatment of Aβ1-42, and 0.117 mg·L-1-0.469 mg·L-1 LSS can inhibit these increased level. CONCLUSION LSS conferred neuroprotection via inhibiting the inflammatory factors expression.

OBJECTIVE To investigate the autophagic effect of the compounds from the Chinese medicinal herbs, Radix polygalae as a potential neuroprotective agent that enhances the clearance of mutant Huntingtin and α- synuclein in PC- 12 cells. METHODS Radix polygalae was extracted with 75% ethanol using refluxing method, and its quality was assayed by UHPLC-TOF-MS. The autophagic effect of Radix polygalae extract, and its major components including polygalacic acid, senegenin and onjisaponin B were investigated using the green fluorescent protein-light chain 3 (GFP-LC3) autophagy detection and Western blot platforms for detecting the autophagic markers, GFP-LC3 puncta formation and LC3 II expression. The degradation of A53T α- synuclein and the inhibition ofα-synuclein oligo merization related to the Parkinson disease were assayed using Western blot and flow cytometer analysis, respectively. While the cytotoxicity and the degradation of the mutant Huntingtin associated with the Huntingtin disease were investigated using MTT method and flow cytometer analysis. RESULTS Radix polygalae ethanol extract and onjisaponin B improved the GFP-LC3 puncta formation and expression of LC3Ⅱ with time and dose manner, and this induction was activated via AMPK-mTOR and Atg 7 gene pathway. Furthermore, the clearance of α-synuclein and mutant Huntingtin was enhanced via autophagy induction with the treatment of Radix polygalae ethanol extract and onjisaponin B. CONCLUSION Findings in the current study provide detailed insights into the protective mechanism of a novel autophagy inducer, onjisaponin B, which is valuable for further investigation as a new candidate agent for modulating neurodegenerative disorders through the reduction of toxicity and clearance of mutant proteins in the cellular level.

OBJECTIVE Lychee seed, a famous traditional Chinese medicine, recently were reported to improve the learning and memory abilities in mice. However, it is still unclear whether lychee seed saponins (LSS) can improve the cognitive function and associated mechanisms. METHODS In present studies, we established the Alzheimer disease (AD) model by injecting Aβ25-35 into the lateral ventricle of rats. Then the spatial learning and memory abilities of LSS- treated rats were evaluated with the Morris water maze, meanwhile the protein expressions of AKT, GSK3β and Tau in the hippo?campal neuron were analyzed by immunohistochemistry and Western blotting. RESULTS The results showed LSS can improve the cognitive functions of AD rats through shortening the escape latency, increasing the number across the platform, platform quadrant dwell time and the percentage of the total distance run platform quadrant. The protein expression of AKT was significantly up-regulated and that of GSK3β and Tau were decreased remarkably in the hippocampal CA1 area. CONCLUSION Our study is the first to show that LSS significantly improve the cognitive function and prevent hippocampal neuronal injury of the rats with AD by activation of the PI3K/AKT/GSK3β signaling pathway, suggesting LSS may be developed into the nutrient supplement for the treatment of AD.

BACKGROUNDThe vessel healing in patients with coronary artery aneurysms (CAA) that form after drug-eluting stent (DES) implantation is not clear. This study aims to assess the vessel healing in patients with CAA formation after DES implanation.

METHODSFrom June 2008 to August 2011, follow-up coronary angiography was conducted on 1160 patients who underwent percutaneous coronary intervention (PCI). The average period of follow-up was about (18.95 ± 13.05) months. A total of 175 patients who underwent DES implantation into de novo lesions and who underwent coronary angiography and optical coherence tomography (OCT) examination during follow-up were identified. Patients were divided into the CAA group (n = 31) and non-CAA group (n = 144) based on the results of the coronary angiography. The cardiac events including angina and acute myocardial infarction were noted; in addition, the neointimal thickness and the frequency of strut malapposition and strut uncoverage were also noted.

RESULTSA greater proportion of incomplete neointimal coverage (17.17% vs. 1.90%, P < 0.001) and strut malapposition (18.20% vs. 1.38%, P < 0.001) were observed in the CAA group. The neointimal thickness in the CAA group was significantly thinner than that in the non-CAA group ((146.6 ± 94.8) µm vs. (192.5 ± 97.1) µm, P < 0.001), as detected via OCT. Patients with CAA formation had a higher frequency of cardiac events including angina pectoris (25.81% vs. 6.25%, P = 0.001) and acute myocardial infarction (9.68% vs. 0.13%, P = 0.002) and thrombosis (16.13% vs. 0.69%, P < 0.001). The longitudinal length of the CAA in the cardiac event group was significantly longer than in the no cardiac event group ((20.0 ± 9.07) mm vs. (12.05 ± 5.38) mm, P = 0.005).

CONCLUSIONCAA formation after DES implantation is frequently associated with cardiac events as a result of stent malapposition and incomplete neointimal coverage.

Adult ; Aged ; Coronary Aneurysm ; diagnosis ; Drug-Eluting Stents ; adverse effects ; Humans ; Middle Aged ; Neointima ; diagnosis ; Percutaneous Coronary Intervention ; adverse effects ; Tomography, Optical Coherence ; methods

Objective To study the single nucleotide polymorphisms (SNP) characteristics of Yersinia pestis strains from different natural foci in China.Methods Genome-wide comparison was done to find SNP sites by the Mummer program among 9 Yersinia pestis genome which was downloaded from NCBI.Then 13 genic fragments including 19 SNP sites were amplified by PCR and sequenced in 133 Yersinia pestis strains,and the results were cluster analyzed with the BioNumerics software.Results Three thousand seven hundred and eighty sequence variation sites were found by genome-wide comparison.Using the different combinations of SNP sites,UPGMA cluster analysis revealed obvious geographic regional and eco-aggregation characteristics of Yersinia pestis strains isolated from China.Conclusions As relatively stable genetic markers,SNP can better reflect the genome characteristics of Yersinia pestis in different plague natural foci of China.

To develop a stable cell line that could express the RSV NS1, the full-length RSV NS1 gene was generated by RT-PCR amplification from respiratory syncytial virus. NS1 gene was ligated with pBABE-puro to construct the recombinant retroviral expression plasmid pBABE-NS1, which was cotransfected into 293FT packaging cells with PIK packaging plasmid by calcium phosphate co-precipitation. The supernatant of 293FT was collected to infect HEp-2 cells, the resulting cell clones stably expressing NS1 were screened by puromycin. Using QPCR, CPE staining method and indirect immunofluorescence assay, the expression of NS1 at both gene and protein levels was identified. The recombinant plasmid pBABE-NS1 was identified by EcoRI and BamHI endonuclease digestion and the sequence analysis. QPCR results showed that the NS1 gene amplification in HEp-2-NS1 cells was 8483 fold higher than that in HEp-2 cells. Although the exogenous interferon was added, all cells were destroyed after 48 hours post infection using CPE staining method, showing that HEp-2-NS1 cells remained sensitive to the VSV virus. The results of RT-PCR and indirect immunofluorescence assay showed that the NS1 gene in HEp-2 cells could not only transcribe mRNA, but also express NS1 protein steadily. We had successfully established HEp-2-NS1 cell lines with stable expression of respiratory syncytial virus non-structural protein NS1.
Cell Line, Transformed ; HEK293 Cells ; Humans ; Recombinant Proteins ; biosynthesis ; genetics ; Respiratory Syncytial Viruses ; genetics ; Viral Nonstructural Proteins ; biosynthesis ; genetics

BACKGROUNDIn general, percutaneous coronary intervention (PCI) relieves vessel stenosis by implantation of a stent, however, the relationship between plaque characteristics and response after stenting is not clear.

METHODSWe enrolled 68 patients (68 vessels) with diagnosed unstable angina pectoris that prospectively underwent PCI and an optical coherence tomography (OCT) examination was done before and after stenting. Coronary plaques were classified as fibrous, lipid-rich and calcified plaque according to OCT examination, and fibrous cap thickness, lumen eccentricity, stent expansion, stent malapposition, tissue prolapse, thrombosis, dissection and stent symmetry were noted.

RESULTSThe frequency of prolapse was higher in lipid-rich plaques than fibrous plaques (85% vs. 40%, P < 0.001). Dissection most often occurred in fibrous plaque compared with lipid-rich and calcified plaques (60% vs. 32% vs. 29%, P < 0.001). The frequency of stent strut malapposition in calcified plaques was higher than firous and lipid-rich plaques (71% vs. 40% vs. 5%, P < 0.001). In-stent micro-thrombosis was detected only in lipid-rich plaques, with a frequency of 37% (15/41). The risk factors of micro-thrombosis after PCI were cap thickness (OR 0.903, 95%CI 0.829 - 0.985), lumen eccentricity (OR 1.147, 95%CI 1.012 - 1.30), and stent length (OR 1.495, 95%CI 1.032 - 2.166).

CONCLUSIONPlaque response after PCI is associated with its characteristics, and of those, micro-thrombosis after stenting in lipid-rich plaques was the most significant finding and can be predicted.

Adult ; Aged ; Aged, 80 and over ; Angioplasty, Balloon, Coronary ; methods ; Coronary Artery Disease ; therapy ; Drug-Eluting Stents ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Tomography, Optical Coherence ; methods

In present study, we investigated the mechanism of regulating HIF-1alpha expression by hydroxysafflor yellow A (HSYA) in Eahy 926 cell line under 1% O2 hypoxia. Eahy 926 cells were incubated with HSYA (100, 10 and 1 micromol x L(-1)) under hypoxia for the indicated time after treatment. Cell proliferation rate was detected using MTT assays. VHL and p53 location and protein expression were analyzed by immunocytochemical stain. HIF-1alpha, VHL and p53 mRNA expression were detected by RT-PCR. Protein expression of HIF-1alpha, VHL and p53 were assayed by Western blotting method. HSYA at 100 micromol x L(-1) increased Eahy 926 cells proliferation rate under hypoxia. HIF-1alpha mRNA and protein expression were up-regulated in the presence of HSYA. VHL, p53 mRNA and protein expression decreased significantly after 8 hours of treatment under hypoxia. HSYA protected Eahy 926 cells from hypoxia, and up-regulated HIF-1alpha expression partially via its inhibition of VHL and p53 expression.
Carthamus tinctorius ; chemistry ; Cell Hypoxia ; Cell Line ; Cell Proliferation ; drug effects ; Chalcone ; analogs & derivatives ; isolation & purification ; pharmacology ; Endothelial Cells ; cytology ; metabolism ; Flowers ; chemistry ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; biosynthesis ; genetics ; Plants, Medicinal ; chemistry ; Quinones ; isolation & purification ; pharmacology ; RNA, Messenger ; metabolism ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics ; Umbilical Veins ; cytology ; Up-Regulation ; Von Hippel-Lindau Tumor Suppressor Protein ; biosynthesis ; genetics

OBJECTIVETo recommend the successive "Z" incision and skin flaps as a method for repairing the wound in penile lengthening procedures.

METHODSWe performed penile lengthening surgery by successive "Z" incision for 5 patients complaining of short penis, who ranged in age from 16 to 34 years (22.4 +/- 7.2), and 3 of whom had a history of urethroplasty, circumcision and penile reconstruction, respectively. The superficial suspensory ligament and part of the deep suspensory ligament of the penis were exposed and severed to release the penis, and the "Z" skin flaps sutured in a tensionless state. Drainage was necessitated by exudation and the catheter removed in 24-48 hours. The penis was wrapped up by an elastic bandage, and the stitches taken out 8-10 days after the operation.

RESULTSWe achieved a mean erectile length of 8.4 cm (range 7.8-9.2 cm) after the operation, as compared with 4. 8 cm (range 4.0-5.8 cm) before the surgery. Two students of the patients obviously became active and confident. No penile contraction was noted during the 12-48 months follow-up, and both the patients and their family members were satisfied with the outcomes.

CONCLUSIONCompared with the V-, M- and Z-incision, the successive "Z" incision and skin flaps can yield longer penile length, solve the problem of insufficient skin flap to cover the wound and reduce such complications as skin necrosis and infection. It is well worth recommending for patients complaining of short penis because of penile abnormality, post-operation scar on the penis or circumcision.

Adolescent ; Adult ; Follow-Up Studies ; Humans ; Male ; Penis ; abnormalities ; surgery ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; Surgical Flaps ; Treatment Outcome