Purpose　To study the proteins expression of genes related to apoptosis of retinal cells in development of human fetus．　Methods　Fifty cases of retinas of human fetus aged from 12 to 38 weeks were collected and paraffin embedded sections were made． Immunohistochemical method was used．　Results　Fas protein was expressed by cells of ganglion cell layer, inner and outer nuclear later, which were just formed on 16th week． It was not expressed until 38th week, Fas(+) staining appeared in layers of retina． Fas-L(+) staining was detected in cells of layers of retina on 26th week and the positive staining located in ganglion cell layer on 32th week． Neuronal fiber layer was Fas-L positive． Bax positive staining was detected on 8th week． Bax positive nucleus were observed mainly in GCL and ONL on 16th week． It was in INL on 24th week and in Müller cells inner terminates on 26th week． After this time, all cells of retina were bax immune negative staining． Bcl-2(+) staining appeared in differentiating neuroblastic layer on 16th week． Beginning on 24th week, bcl-2 (+) staining was observed in glial cells of GCL and inner terminates of Müller cell．　Conclusion　Apoptosis of developing retinal cell may be Fas/Fas-L independent and bax may be involved in apoptosis of the cells．

Objective To investigate the clinical features of multifocal choroiditis (MC) and guide the diagnosis and treatment. Methods Retrospective analysis of clinical data of 18 MC cases (28 eyes) who were diagnosed through fluorescein angiography (FFA) or indocyanine green angiography (ICGA) and fundus characteristics. Results Multiple round to oval lesions scattered throughout the posterior pole and peripheral areas of ocular fundi of all of the 28 eyes(binocular in 10 and monocular in 8) were found. Active focal lesions of ocular fundi were seen in 8 patients and inactive lesions in 10 patients. active and 10 cases were inactive. Choroidal neovascularization(CNV) in macular area was found in 7 patients. The images of FFA of the legions showed hypofluorescence in the early phase, with late leakage and gradual staining or window is defect in the late phase. Conclusions MC is a rare disease and often misdiagnosed to other disease and FFA helpful in diagnosis.

Objective To observe the expression of p53, bcl 2 genes, vascular endothelial cell growth factor (VEGF), basic fibroblast growth factor (bFGF), insulin like growth factor I (IGF I), and the receptors of these factors of retinal vascular endothelial cells (VECs) of 1 to 20 week diabetic rats, and the relationship between the expressions and cell cycle arrest. Methods Retinal sections of diabetic rats induced by alloxan were immunohistochemically stained and observed by light microscopy (LM) and electron microscopy (EM). Dot blotting and Western blotting were used to determine the expression of mRNA, proteins of p53 and bcl 2. Results Under LM, immunohistochemical positive expression of p53 and bcl 2 were found on the vessels of ganglion cell layer and inner nuclear layer of retinae of 8 to 20 week diabetic rats; under EM, these substances were observed depositing in VECs. The retinal VECs also expressed VEGF, bFGF, IGF I and their receptors. There was no positive expression of other cell types in these retinae, all cell types of retinae in control group, or all cells of retinae of diabetic rats with the course of disease of 1 to 6 weeks. The result of dot blotting revealed that retinal tissue of 20 week diabetic rat expressed p53 and bcl 2 mRNA, and the result of Western blotting revealed that they also expressed p53 and bcl 2 proteins. But retinal tissues of control group did not. Positive expression of bax was not found in the retinae in control group or 1 to 20 week diabetic rats. Conclusion p53, bcl 2 may introduce cell cycle arrest of VECs of retinae in 8 to 20 week diabetic rats. High glucose might stimulate the expression of VEGF, bFGF, IGF I and their receptors, and the growth factors may keep VECs surviving by self secretion.

Objective To investigate the characteristics of early changes in serum IL-17 and IFN-γ levels in elderly patients with acute myocardial infarction (AMI).Methods 70 hospitalized elderly patients with AMI and 35 healthy people were selected.Serum level of interleukin-17 (IL 17) and interferon-γ (IFN-γ) were assayed by enzyme linked immunosorbent assay (ELISA).Results Serum levels of IL17 and IFN-γ showed increasing trends in elderly patients with AMI as compared with that in control group,but there were no significant differences between the two group [(35.73 30.28) pg/ml vs.(28.70±17.12) pg/ml,(15.29±14.64) pg/ml vs.(11.38±10.10) pg/ml,t=0.144 and 0.138,P=0.365 and 0.377].There were correlations between serum IL-17 and IFN γ levels in patients with AMI and controls(r=0.936 and 0.989,both P=0.00).Serum levels of IL-17 or IFN-γhad no significant differences between AMI patients with well and poor prognosis [(35.43± 34.36) ng/L vs.(36.11±30.16) ng/L,(13.90±13.98) ng/L vs.(15.99±14.14) ng/L,U=0.266 and 0.166,P=0.687 and 0.668].Conclusions Serum IL-17 level has an increasing trend in AMI patients within 24h,but has no statistical significant.Serum IL 17 level has a significantly positively correlation with serum IFN γ level in the elderly,but serum levels of IL-17 or IFN γ have no significant correlations with short term prognosis in elderly patients with AMI.

ObjectiveTo explore the relationship among 25-hydroxyvitamin D,serum calcium,calcium-sensing receptor,and breast cancer. Methods The expressions of calcium-sensing receptor (CaSR) in primary breast cancer,breast benign tumors,and normal breast tissue beside tumors were determined by immunohistochemistry S-P method as well as the concentration of serum 25 (OH) D and serum calcium in breast cancer and breast benign tumors by Enzyme-linked immunosorbent assay (ELISA),Tribromoarsenazo Ⅲ method.ResultsSerum 25 (OH) D level of breast cancer was significantly lower than the breast benign tumors [ (34.13 ± 14.14) nmol/L vs (50.29 ± 25.65 ) nmol/L,t =2.870,P =0.001 ].Serum level of 25 ( OH ) D in lymph node metastasis positive patient was lower than that in negative group [ (30.8 ± 9.71 ) nmol/L vs (43.7 ± 23.59) nmol/L,t =2.467,P =0.021 ].The positive expression of CaSR in breast cancer(88.9％ )was higher than breast benign tumors(60％,x2 =6.717,P ＜ 0.01 ) and normal breast tissue beside tumors (60％,x2 =5.628,P ＜ 0.05 ).ConclusionsConcentration of serum 25 (OH)D and expression of calcium-sensing receptor in the tissues may be associated with occurrence,development and prognosis of breast cancer.

A double-molecular beacons (DMB) based assay was developed for porcine circovirus 2(PCV2) detection. Two single-stranded DNA molecular beacons which could specifically hybridize with PCV2 genome DNA respectively in different sequence were designed according to the characteristics of the PCV2 genome sequences. The fluorescence signal was amplified 80 times by DMB, which was 2-4 times higher than that of single molecular beacon. Under the optimal conditions of 10 mmol/L MgCl2 , 20 mmol/L Tris-HCl (pH=8. 0), 40 ℃ and 30 min incubation time of DNA with DMB, the enlargement factor was increased linearly with DNA concentration over the range from 2 nmol/L to 200 nmol/L, with a detection limit of 1 nmol/L. The method was applied to detect PCV2 in genome of 18 swine fever samples and 8 PCV2 positive cases were found, which were confirmed by PCR method.

Objective:To understand the genotyping and molecular epidemiological characteristics of Norovirus (NoV) GⅡ in adult acute gastroenteritis outpatients in Shanghai from 2015 to 2016.Methods:A total of 912 stool specimens from adult patients with acute diarrhea from March 2015 to December 2016 (431 in 2015 and 481 in 2016) were collected. The Norovirus GⅡ type was detected by one-step quantitative reverse transcription PCR assay and the RdRp region of open reading frame (ORF) 1 and 5′end of ORF2 were amplified. The region was sequenced and classified.Results:From March 2015 to December 2016, NoV GⅡ were detected in 17.76% (162/912) of the samples, 15.08% (65 /431) in 2015 and 20.17% (97/481) in 2016. Based on sequence analysis of the RdRp and capsid sequences, 145 identified NoV strains were divided into 10 genotypes: GⅡ.Pe_GⅡ.4 Sydney 2012 (60), GⅡ.P17_GⅡ.17 (45), GⅡ.P16_GⅡ.13 (10), GⅡ.P12_GⅡ.3 (10), GⅡ.P7_GⅡ.6 (9), GⅡ.P21_GⅡ.21 (5), GⅡ.P16_GⅡ.4 Sydney 2012 (2), GⅡ.Pe_GⅡ.17 (2), GⅡ.P2_GⅡ.2 (1), and GⅡ.P7_GⅡ.14 (1).Conclusions:The main epidemic NoV GⅡ genotype in Shanghai was GⅡ.P17_GⅡ.17 in the spring of 2015. GⅡ.Pe_GⅡ.4 Sydney 2012 and GⅡ.P17_GⅡ.17 were identified as predominant genotypes during the winter of 2015 and spring of 2016. The most common genotype was GⅡ.Pe_GⅡ.4 Sydney 2012 in autumn of 2016. Continuous NoV outbreak surveillance is important for identifying changing trends in genotype distribution and emerging new strains.

Objective:To construct mitochondrial antiviral signaling ( MAVS) gene or Nuclear factor kappa B1 ( NFκB1) gene knockout Madin-Darby canine kidney (MDCK) cells, and identify the cell growth and proliferation characteristics of influenza virus (Flu) in these cells. Methods:MAVS or NFκB1 knockout MDCK cells were established using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) technique. Flu was inoculated into these cells, then the hemagglutination (HA) titers and TCID 50 were determined and compared with the original MDCK cells. Results:MDCK cells knocked out of MAVS (MDCK- MAVS-) or NFκB1 (MDCK- NFκB1 -) were obtained stably. The two cells were both adherent epithelioid cells as well as wild type MDCK cells. The HA titers showed no obvious difference among these cells after inoculating Flu. Nevertheless, the TCID 50 titers were respectively increased 9.5 times in MDCK- MAVS-cell culture and 10 times in MDCK- NFκB1 - cell culture, compared to the wild type MDCK cells. Conclusions:The infectivities of Flu were both increased in MDCK- MAVS-cells and MDCK- NFκB1 - cells, illustrating their potential in Flu culture.

Infectious bronchitis virus (IBV) poses a severe threat to the poultry industry and causes heavy economic losses worldwide. Vaccination is the most effective method of preventing infection and controlling the spread of IBV, but currently available inactivated and attenuated virus vaccines have some disadvantages. We developed a chimeric virus-like particle (VLP)-based candidate vaccine for IBV protection. The chimeric VLP was composed of matrix 1 protein from avian influenza H5N1 virus and a fusion protein neuraminidase (NA)/spike 1 (S1) that was generated by fusing IBV S1 protein to the cytoplasmic and transmembrane domains of NA protein of avian influenza H5N1 virus. The chimeric VLPs elicited significantly higher S1-specific antibody responses in intramuscularly immunized mice and chickens than inactivated IBV viruses. Furthermore, the chimeric VLPs induced significantly higher neutralization antibody levels than inactivated H120 virus in SPF chickens. Finally, the chimeric VLPs induced significantly higher IL-4 production in mice. These results demonstrate that chimeric VLPs have the potential for use in vaccines against IBV infection.
Animals ; Antibodies, Viral/blood ; *Chickens ; Chimera/genetics/immunology ; Coronavirus Infections/prevention & control/*veterinary/virology ; Female ; *Immunity, Innate ; Infectious bronchitis virus/genetics/*immunology ; Influenza A Virus, H5N1 Subtype/genetics/immunology ; Injections, Intramuscular/veterinary ; Mice ; Mice, Inbred BALB C ; Neuraminidase/genetics ; Poultry Diseases/*prevention & control/virology ; Recombinant Fusion Proteins/genetics/immunology ; Spike Glycoprotein, Coronavirus/genetics/*immunology ; Vaccines, Synthetic/administration & dosage/genetics/immunology ; Vaccines, Virus-Like Particle/administration & dosage/genetics/*immunology ; Viral Proteins/genetics

OBJECTIVETo determine the levels of urinary soluble intercellular adhesion molecule-1 (sICAM-1) and vascular cellular adhesion molecule-1 (VCAM-1) in patients with lupus nephritis (LN), and explore their correlation with renal disease activity.

METHODSUrine samples were collected from 92 renal biopsy-proven LN patients and 20 healthy controls. Renal disease activity was determined according to the ISN/RPS 2003 Revised Classification of Lupus Nephritis. The urine levels of sICAM-1 and VCAM-1 were detected by enzyme-linked immunosorbent assay, and the expressions of intrarenal ICAM-1 and VCAM-1 were evaluated by immunohistochemisty staining.

RESULTSUrinary sICAM-1 and VCAM-1 levels were elevated in LN patients compared with the controls. Significantly higher levels of urinary sICAM-1 and VCAM-1 were found in patients with active LN, who had also significantly increased intrarenal expressions of ICAM-1 and VCAM-1 compared with patients in remission. A strong positive correlation was noted between intrarenal expression and urine levels of ICAM-1 and VCAM-1. The receiver-operating characteristic (ROC) curve of urine sICAM-1 showed tan area under ROC curve of 0.874 for all participants in the test. A cutoff of 1095.00 pg/mg creatinine yielded a good sensitivity (0.945) and specificity (0.789). The ROC curve of urine VCAM-1 showed an area under ROC of 0.882 for all the participants, and a cutoff of 898.11 pg/mg creatinine yielded a good sensitivity (0.982) and specificity (0.667).

CONCLUSIONUrine sICAM-1 and VCAM-1 levels are positively correlated with their intrarenal expressions and reflect the activity of the nephritis, and therefore they may serve as potential biomarkers for early diagnosis of active LN.

Biomarkers ; urine ; Case-Control Studies ; Early Diagnosis ; Enzyme-Linked Immunosorbent Assay ; Humans ; Intercellular Adhesion Molecule-1 ; urine ; Kidney ; metabolism ; Lupus Nephritis ; diagnosis ; urine ; ROC Curve ; Sensitivity and Specificity ; Vascular Cell Adhesion Molecule-1 ; urine