Immunological microenvironment alterations in follicles of women with proven severe endometriosis undergoing in vitro fertilization.

The purpose of this study was to test the hypothesis that different cytokine profiles may exist in the follicular fluid of endometriosis (EM) patients undergoing in vitro fertilization (IVF), as these differences may provide insights into the pathogenesis of the disease. This was a cross-sectional study conducted at the reproductive center of a medical university hospital. The study included 49 patients receiving IVF. 20 infertile women with proven EM and 29 women without diagnosed EM (control group) were evaluated. Follicular fluid (FF) and serum were collected at the time of follicle aspiration and the concentrations of 38 cytokines were determined by multiplexed immunoassay. The results indicated that the levels of IL-4, IL-13, IL-3 and IL-1α were significantly increased in the FF of women with EM, while levels of IFN-γ, IL-17A, MDC and MIP-1α were decreased compared with in the control subjects. In conclusions, the immune microenvironment of the FF in patients with EM is altered. This may contribute to the pathologic mechanism responsible for the poor outcome of IVF in patients with EM.

The Protective Effects of the A/ZJU01/ PR8/2013 Split H7N9 Avian Influenza Vaccine Against Highly Pathogenic H7N9 in BALB/c Mice.

BACKGROUND/AIMS: Since the first case of novel H7N9 infection was reported, China has experienced five epidemics of H7N9. During the fifth wave, a highly pathogenic H7N9 strain emerged. In order to assess whether the H7N9 vaccine based on A/Zhejiang/DTID-ZJU01/2013(H7N9) was effective in protecting against highly pathogenic H7N9, we conducted this study. METHODS: Groups of mice were immunized twice by intraperitoneal injection with 500 µl of either split vaccine alone or MF59-adjuvanted vaccine. Serum was collected 2 weeks after the second vaccine booster. The hemagglutinin inhibition test was conducted on vaccine seed and highly pathogenic H7N9 to evaluate the neutralization of highly pathogenic H7N9. We also immunized mice and challenged them with highly pathogenic H7N9. Mice were observed for illness, weight loss, and death at 1 week and 2 weeks post-infection. Then, the mice were sacrificed and lungs were removed. Antibody responses were assessed and pathological changes in the lung tissue were evaluated. RESULTS: The ability of serum to neutralize highly pathogenic H7N9 was reduced. In mice, highly pathogenic H7N9 was more virulent than A/Zhejiang/DTID-ZJU01/2013(H7N9). After challenge with highly pathogenic H7N9, all mice died while mice challenged with A/Zhejiang/DTID-ZJU01/2013(H7N9) all recovered. The A/ZJU01/PR8/2013 split H7N9 avian influenza vaccine was able to protect against infection with highly pathogenic H7N9 in mice, with or without MF59. Moreover, H7N9 vaccine adjuvanted with MF59 produced high antibody levels, which lead to better protection. CONCLUSIONS: The A/ZJU01/PR8/2013 split H7N9 avian influenza vaccine based on A/Zhejiang/DTID-ZJU01/2013(H7N9) is effective in protecting against highly pathogenic H7N9. H7N9 vaccine adjuvanted with MF59 offers better protection against infection with highly pathogenic H7N9. In order to make the H7N9 vaccine applicable to humans, further clinical trials are required to evaluate MF59 adjuvanted vaccine. Meanwhile, the vaccine strain should be updated based on the highly pathogenic H7N9 gene sequence.

Process optimization for the rapid production of adenoviral vectors for clinical trials in a disposable bioreactor system.

Recombinant adenoviral (Ad) vectors are highly efficient gene transfer vectors widely used in vaccine development and immunotherapy. To promote the industrial application of Ad vectors, studies focusing on reducing the cost of manufacturing, shortening the preclinical research period, and improving the quality of products are needed. Here, we describe a highly efficient and economical process for producing Ad vector in a novel, single-use bioreactor system suitable for clinical trials. A mini-bioreactor was used for parameter optimization and development of medium replacement protocols for Ad5-GFP production before scale-up. HEK293 cell culture and virus infection were monitored in a disposable AmProtein Current Perfusion Bioreactor and Bioflo310 bioreactor using optimized parameters and medium replacement protocols. The total cell number increased from 2.0 × 109 to 3.2 × 1010 after 6 days of culture. The total number of viral particles obtained in a single batch was 1.2 × 1015. These results demonstrate the efficiency and suitability of this system for Ad vector production for research and GMP applications.

High-throughput sequencing identifies HIV-1-replication- and latency-related miRNAs in CD4+ T cell lines.

MicroRNAs are potent gene expression regulators involved in regulating various biological processes, including host-pathogen interactions. In this study, we used high-throughput sequencing to investigate cellular miRNA signatures related to HIV-1 replication and latent infection in CD4+ T cell lines, which included HIV-1-replicating H9/HTLV-IIIB, HIV-1-latently-infected CEM-Bru cells, and their parental uninfected H9 and CEM-SS cells. Relatively few miRNAs were found to be modulated by HIV-1 replication or latent infection, while the cell-lineage-specific miRNA difference was more pronounced, irrespective of HIV-1 infection. In silico analysis showed that some of our HIV-1 infection-regulated miRNA profiles echoed previous studies, while others were novel. In addition, some of the miRNAs that were differentially expressed between the productively and latently infected cells seemed to participate in shaping the differential infection state. Thus, the newly identified miRNA profiles related to HIV-1 replication and latency provide information about the interplay between HIV-1 and its host.

Efficacy of oseltamivir-peramivir combination therapy compared to oseltamivir monotherapy for Influenza A (H7N9) infection: a retrospective study.

BACKGROUND: Since the novel H7N9 avian influenza outbreak occurred in China in 2013, neuraminidase inhibitors (NAIs) such as oseltamivir and peramivir have been used as first-line drugs to treat the influenza virus infection. This study aimed to compare the efficacy of oseltamivir-peramivir combination therapy versus oseltamivir monotherapy. METHODS: A retrospective study of 82 H7N9 confirmed patients was conducted by reviewing medical charts at the First Affiliated Hospital of ZheJiang University in China from April 1, 2013 to Feb 28, 2014. The patients' clinical information was collected systematically, and we compared the virology and clinical data between oseltamivir monotherapy group (43 patients) and oseltamivir-peramivir combination group (39 patients). RESULTS: The median duration from NAIs administration to H7N9 virus-negative in oseltamivir monotherapy group and oseltamivir-peramivir combination group was 6.50 and 7.00 days (p >0.05), respectively. The median decline of Day 2 to Day 0 (initiation of NAIs therapy) viral load was 0.00 and 0.69 log10 copies/µl (p >0.05) respectively in the monotherapy vs. combination therapy groups. The incidence of new Acute Respiratory Distress Syndrome during NAI administration was 63.89 and 77.78 % (p >0.05); while the mortality rates were 25.58 and 43.59 % (p >0.05) in the oseltamivir group vs. oseltamivir-peramivir group. CONCLUSIONS: Our results suggest that in adults with H7N9 virus infection, the use of oseltamivir-peramivir combination therapy was not superior to oseltamivir monotherapy.

[The effect of OPTC-shRNA on the bovine retinal pigment epithelial cells and hyalocytes co-culture collagen gel contraction system].

OBJECTIVE: To investigate the OPTC-shRNA inhibiting effect on the opticin expression by the bovine hyalocytes and retina pigment epithelial (RPE) cells co-culture collagen gel contraction system. METHODS: Experimental study. The OPTC-shRNA expression vector was designed and transfected into bovine RPE cells cultured in vitro. The relative expression and the inhibition rate of the opticin protein were measured by Western blot on days 3, 5 and 7. An in vitro cells co-culture bovine type I collagen gel contraction assay was constructed consisting of the hyalocytes and RPE cells. Six groups were established in this experiments:OPTC-shRNA plasmid transfected RPE cells and hyalocytes (group A), empty plasmid transfected RPE cells and hyalocytes (group B), non-transfected RPE cells and hyalocytes (group C), non-transfected RPE cells (group D), only hyalocytes (group E), and no cells (group F). The collagen gel contractile activities of these groups were compared by One-way ANOVA, SNK-q tests and regression analysis;and the influence of the hyalocytes density variance on the collagen gel contraction in groups A, B and C were also analyzed. RESULTS: The OPTC-shRNA expression vector with significant inhibition effect was constructed and transfected into bovine RPE cells successfully. The results of Western blot analysis showed that the inhibitory rates on the opticin expression on days 3, 5 and 7 were (83.91 ± 2.88), (84.71 ± 4.27) and (82.85 ± 2.72)%, respectively. Furthermore, the differences among days 3, 5 and 7 were insignificant (F = 1.15, P > 0.05). On day 3, the gel contraction rates for the sub-groups with various hyalocytes densities (2×10(7), 1×10(8) and 5×10(8)/L) in groups A, B and C were: group A: (23.52 ± 2.08), (56.00 ± 1.02), (61.62 ± 1.73)%; group B: (16.56 ± 2.01), (36.41 ± 1.33), (49.56 ± 1.75)%; group C: (15.75 ± 1.37), (37.45 ± 1.14), (48.45 ± 1.97)%. The gel contraction rates for groups D and E were (12.18 ± 0.95)% and (10.95 ± 0.93)%, respectively; no gel contraction was observed in group F. Pairwise comparisons of the gel contraction rates were performed by SNK-q test among groups A, B and C for various hyalocyte densities. In the 2×10(7)/L cell density group, the differences between groups A and B or C were significant (q = 11.38, 2.72, respectively, P both < 0.05), the differences between B and C were insignificant (q = 1.34, P > 0.05). In the 1×10(8)/L cell density group, the differences between groups A and B or C were significant (q = 8.83, 46.22, respectively, P both < 0.05), the differences between B and C were insignificant (q = 1.34, P > 0.05). In the 5×10(8)/L cell density group, the differences between groups A and B or C were significant (q = 48.83, 46.22, respectively, P both < 0.05), the differences between groups B and C were insignificant (q = 1.74, P > 0.05). Pairwise comparisons of the sub-groups with different hyalocyte densities in groups A, B and C (comparisons of 2×10(7)/L and 1×10(8)/L, 2×10(7)/L and 5×10(8)/L, 2×10(7)/L and 2×10(7)/L, respectively), the differences were all significant (group A:q = -55.97, -65.66, -9.69, respectively; group B: q = -34.53, -57.41, -22.88, respectively; group C: q = -41.94, -63.19, -21.25, P all < 0.05). Furthermore, the regression analysis was performed between the hyalocyte density and the collage gel contraction rates in each group. The results showed that there was a positive correlation between the gel contraction rates of the co-culture collagen gel contraction system and its hyalocyte density (groups A, B, C: r = 0.919, 0.981, 0.937, respectively, P all < 0.05). Pairwise comparison of groups D and E, D and F, E and F by SNK-q test revealed q = 54.87, 49.33, 5.54, respectively, P all < 0.05. CONCLUSION: Opticin is capable of regulating the contraction of bovine hyalocytes and RPE cells co-culture collagen gel.

[Spectral domain optical coherence tomography to assess the association between the inner caliber of large retinal vessel and the primary hypertension].

OBJECTIVE: To assess the inner caliber of large retinal vessel quantitatively using spectral domain optical coherence tomography (SD-OCT) and to reveal the association between changes in the inner caliber of large retinal vessel and the primary hypertension. METHODS: A retrospective case-control study was carried out including 215 cases (with primary hypertension) and 210 controls (without primary hypertension) admitted to our hospital since 2009 and all the cases and controls were grouped according to age. SD-OCT was performed to assess the inner caliber of large retinal vessel quantitatively including retinal artery inner caliber (RAIC), retinal vein inner caliber (RVIC) and retinal arterio-venous inner caliber ratio (RAVICR). The differences in the inner caliber of large retinal vessel between the cases and the controls in each age groups were analyzed by t test. In all cases, multiple comparisons were performed according to their blood pressure level by SNK test of one way ANOVA. The RAVICR was also correlated with the following relevant determinants via multiple stepwise regression analysis: age, diastolic and systolic blood pressure. RESULTS: In each age group of cases (< 40, 40 to 49, 50 to 59, 60 to 69, ≥ 70 years), the values of RAIC were (93.0 ± 6.3), (86.2 ± 6.1), (84.5 ± 5.1), (84.0 ± 5.5), and (81.7 ± 5.4) µm respectively, and the values of RVIC were (129.4 ± 5.8), (130.7 ± 6.5), (129.6 ± 5.4), (132.2 ± 6.4), and (131.6 ± 5.1) µm respectively, and the values of RAVICR were (0.720 ± 0.07), (0.661 ± 0.06), (0.653 ± 0.04), (0.637 ± 0.06), and (0.621 ± 0.05) µm respectively. Compared with controls, RAIC (t = -4.813, -10.893, -15.689, -8.811, and -10.151 respectively; P < 0.05) and RAVICR (t = -3.276, -8.654, -13.470, -7.801, and -9.210 respectively; P < 0.05) were significantly decreased in each age group of cases. Multiple comparisons were made among each systolic and diastolic pressure groups in all cases. In systolic groups, difference of RAIC or RAVICR were significant (SNK test)between 140 to 149 mm Hg (1 mm Hg = 0.133 kPa) and 170 to 179 mm Hg group (q = 9.46, 10.61; P < 0.05), 140 to 149 mm Hg and ≥ 180 mm Hg group (q = 11.03, 13.98; P < 0.05), 150 to 159 mm Hg and 170 to 179 mm Hg group (q = 8.13, 8.82; P < 0.05), 150 to 159 mm Hg group and ≥ 180 mm Hg group (q = 9.01, 9.97; P < 0.05). In diastolic groups, difference of RAIC or RAVICR were significant (SNK test) between 90 to 99 mm Hg and 100 to 109 mm Hg group (q = 6.79, 5.95;P < 0.05), 90 to 99 mm Hg and ≥ 110 mm Hg group (q = 9.72, 10.21; P < 0.05), 100 to 109 mm Hg and ≥ 110 mm Hg group (q = 5.93, 6.07; P < 0.05). RAVICR was associated with the diastolic and systolic blood pressure revealed by the multiple stepwise regression analysis (ANOVA: F = 11.231; Standardized regression coefficient: ß = -0.024, -0.019, respectively; P < 0.05). CONCLUSIONS: Quantitative assessment for the inner caliber of large retinal vessel can be done by SD-OCT. The value of RAI and RAVICR were correlated with diastolic and systolic blood pressure in primary hypertension.

Enhanced Programmed Cell Death Protein 1/Programmed Cell Death Ligand 1 Expression Induced by Severe Influenza A Virus Infection Impairs Host's Antiviral Response.

Emerging research shows that the Programmed Cell Death Protein 1/Programmed Cell Death Ligand 1(PD-1/PD-L1) pathway modulates the antiviral response following influenza A virus (IAV) infection, and there is a need to understand further the role of the PD-1/PD-L1 signaling pathway in IAV infection. BALB/c mice were infected with different types of IAV to establish models of varying degrees of infection (mild and severe). The mice were pretreated with or without a PD-1 antagonist to evaluate the role of the PD-1/PD-L1 pathway in IAV infection. The general activity, degree of weight change, viral titer, pathological damage, protein expression, transcriptome level, and cytokine expression were evaluated in the mice. IAV infection, especially severe infection, induced expression of PD-1 and PD-L1 in the lungs and spleen of the mice at 6 days postinfection. Moreover, the expression level was positively correlated with the degree of pathological damage in the lung. PD-1 antagonists can alleviate weight loss in severely infected mice, reduce the viral load and pathological damage, enhance immune response-related gene expression, and induce the most robust responses of interferon-gamma without inducing an obvious Th1/Th17 response. The PD-1/PD-L1 signaling pathway induced by severe IAV infection seriously impairs the host's antiviral response; thus, blocking this signaling pathway may promote IAV recovery.

Bioinformatic analysis of PD-1 checkpoint blockade response in influenza infection.

BACKGROUND: The programmed cell death 1 (PD-1)/PD-1 ligand 1 (PD-L1) signaling pathway is significantly upregulated in influenza virus infection, which impairs the antiviral response. Blocking this signaling pathway may reduce the damage, lower the virus titer in lung tissue, and alleviate the symptoms of infection to promote recovery. In addition to the enhanced viral immune response, using of immune checkpoint inhibitors in influenza virus infection is controversial, the aim of this study was to identify the key factors and regulatory mechanisms in the PD-1 checkpoint blockade response microenvironment in influenza infection. METHODS: A BALB/c mouse model of influenza A/PR8(H1N1) infection was established then constructed, and whole-transcriptome sequencing including mRNAs, miRNAs (microRNAs), lncRNAs (long noncoding RNAs), and circRNAs (circular RNAs) of mice treated with PD-1 checkpoint blockade by antibody treatment and IgG2a isotype control before infection with A/PR8(H1N1) were performed. Subsequently, the differential expression of transcripts between these two groups was analyzed, followed by functional interaction prediction analysis to investigate gene-regulatory circuits. RESULTS: In total, 84 differentially expressed dif-mRNAs, 36 dif-miRNAs, 90 dif-lncRNAs and 22 dif-circRNAs were found in PD-1 antagonist treated A/PR8(H1N1) influenza-infected lungs compared with the controls (IgG2a isotype control treated before infection). In spleens between the above two groups, 45 dif-mRNAs, 36 dif-miRNAs, 57 dif-lncRNAs, and 24 dif-circRNAs were identified. Direct function enrichment analysis of dif-mRNAs and dif-miRNAs showed that these genes were mainly involved in myocardial damage related to viral infection, mitogen activated protein kinase (MAPK) signaling pathways, RAP1 (Ras-related protein 1) signaling pathway, and Axon guidance. Finally, 595 interaction pairs were obtained for the lungs and 462 interaction pairs for the spleens were obtained in the competing endogenous RNA (ceRNA) complex network, in which the downregulated mmu-miR-7043-3p and Vps39-204 were enriched significantly in PD-1 checkpoint blockade treated A/PR8(H1N1) infection group. CONCLUSIONS: The present study provided a basis for the identification of potential pathways and hub genes that might be involved in the PD-1 checkpoint blockade response microenvironment in influenza infection.

[The effect of vascular endothelial growth factor and hypoxia on the secretion of opticin in retinal pigment epithelium cells].

OBJECTIVE: To evaluate the role of vascular endothelial growth factor or hypoxia on the secretion of opticin in retinal pigment epithelium cells. METHODS: Human RPE cells were cultured, the third to sixth passage of retinal pigment epithelium (RPE) cells were placed in 6-well culture plates at a density of 4 × 10(4)/well. For hypoxia experiment, the cells were cultured under hypoxic condition for different times. For vascular endothelial growth factor (VEGF) experiment, the media was changed to DMEM containing different concentration VEGF (1, 10, 50, 100 µg/L) for 24 h respectively. VEGF mRNA levels were determined by RT-RCR method. The protein content of opticin in RPE cells or culture media was detected by Western blot. Matrix metalloproteinase activity in culture media was analysis by zymography. One way ANOVA was used to test the comparisons between experimental groups and control group. RESULTS: Western blot experiment showed the opticin expression was not changed in RPE cells after hypoxia treatment, however was significantly decreased in culture media. Compared with control group (0.21 ± 0.03). The relative density of VEGF mRNA levels (0.81 ± 0.04, 0.67 ± 0.07) in RPE cells were increased after 12 h or 24 h hypoxia treatment (F = 483.60, P < 0.05). Opticin expression in RPE cells was also remain unchanged after vary concentration VEGF addition treatment (F = 2.16, P > 0.05), the relative density of opticin expression in VEGF conditioned culture medium were 0.65 ± 0.02, 0.52 ± 0.04, 0.23 ± 0.03, 0.30 ± 0.03 respectively, and the difference in culture media was significant compared to control group (0.73 ± 0.04) (F = 141.38, P < 0.05). Zymography indicate a matrix metalloproteinases type 2 digest band, the activities were enhanced with VEGF increasing. The decrease of opticin in culture media after VEGF treatment could be inhibited by low condition of EDTA. CONCLUSION: VEGF and hypoxia have an effect on the on the secretion of opiticin in RPE cells, it may be contributed to the increasing levels of matrix metalloproteinases type 2.

MicroRNA Expression Profiling of Lung Cancer with Differential Expression of the RON Receptor Tyrosine Kinase.

BACKGROUND: The Ron receptor tyrosine kinase (RON) can act as a protooncogene and may play a prominent role in the initiation and development of lung cancer. microRNAs (miRNA) are master regulators of gene expression through direct or indirect regulation, and impact all aspects of cell biology. METHODS: Nonsmall-cell lung cancer (NSCLC) samples and small-cell lung cancer (SCLC) were stratified based on RON expression to identify miRNA profiles associated with RON expression levels, differentially expressed miRNA regulated by RON were screened out, and their biological behavior was analyzed. RESULTS: miRNA expression was most significantly affected by cancer type, and we found 85 miRNAs that were significantly differentially expressed between NSCLC and SCLC. There were 46 miRNAs differentially expressed between high RON expressing NSCLC compared to low RON expressing NSCLC. Biological processes and pathways found to be significantly influenced by RON expression included epithelial-mesenchymal transition (EMT) and activation of the PI3K-Akt and MAPK signaling pathways. CONCLUSIONS: These data may provide the basis for a novel strategy to characterize lung cancer by RON expression and microRNA genotyping.

miR-21-5p promotes lung adenocarcinoma cell proliferation, migration and invasion via targeting WWC2.

OBJECTIVE: Studies have suggested that miR-21-5p and WWC2 are key players in most cancer types, yet the underlying mechanisms in lung adenocarcinoma (LUAD) remain elusive. This study made in-depth research on the two factors-dependent mechanisms underlying LUAD occurrence and development. METHODS: Bioinformatics methods were employed to identify the miRNA and its target gene of interest. In all, 20 pairs of LUAD tumor tissue samples and matched adjacent normal samples along with 5 LUAD cell lines were collected for evaluating the aberrant expression of miR-21-5p and WWC2. Dual-luciferase reporter assay was performed to validate the targeted relationship between miR-21-5p and WWC2. A series of in vitro experiments including colony formation assay, EdU, wound healing assay and Transwell were conducted for assessment of the LUAD cell biological behaviors. In addition, Western blot was carried out to determine the protein expression of epithelial-mesenchymal transition (EMT)-related proteins. RESULTS: miR-21-5p was found to be considerably increased in LUAD tissue and cells relative to that in the adjacent tissue and the human bronchial epithelial cells, whereas WWC2 was significantly decreased. Dual-luciferase reporter assay revealed that miR-21-5p targeted WWC2 and down-regulated its expression. Besides, silencing miR-21-5p or overexpressing WWC2 played an inhibitory role in PC-9 cancer cell proliferation, migration and invasion, but such effect was suppressed when miR-21-5p was overexpressed. Furthermore, Western blot uncovered that WWC2 overexpression impeded the EMT process in LUAD cells. CONCLUSION: miR-21-5p facilitates LUAD cell proliferation, migration and invasion through targeting WWC2, which provides a novel therapeutic target for LUAD treatment.

Patients with chronic hepatitis C receiving sofosbuvir and ribavirin-based treatment, with or without interferon in Zhejiang, China: An observational study.

Hepatitis C virus (HCV) is one of the most important virus as the cause of liver disease in China. The aim of the present study was to explore whether sofosbuvir and ribavirin-based treatment can cure patients with chronic hepatitis C in eastern China. We examined a cohort of HCV-monoinfected patients and 9 patients agreed to participate in our treatment and research. The patients were diagnosed with chronic hepatitis C with or without cirrhosis. Nine patients including 4 female and 5 male met the requirements for selection and were willing to participate in this experiment. Sofosbuvir and ribavirin-based treatment with or without interferon was given to the patients. Viral loads, cytokines, and chemokines were recorded during treatment and after treatment. After 2 weeks of sofosbuvir and ribavirin-based treatment, the viral load of patients decreased to limits of detection. Eight patients were cured. Patients had rapid virological response (RVR) with undetectable viral load at week 4 and sustained virological response (SVR). The interferon-inducible protein-10 (IP-10) decreased after the treatment. However, the patient with cirrhosis failed, as the virus reappeared during SVR4. At the same time, the IP-10 dramatically increased as the relapse of the HCV virus. In summary, the IP-10 has the potential to be the biomarker for the prognostic of HCV.

Longevity of protective immune responses induced by a split influenza A (H7N9) vaccine mixed with MF59 adjuvant in BALB/c mice.

The influenza virus is a serious threat to public health worldwide. A novel avian influenza A (H7N9) virus with a mortality rate of approximately 30% has been identified as an unusually dangerous virus for humans by the World Health Organization. Pathogenic H7N9 continue to represent a public health concern, and several candidate vaccines are currently in development. We generated candidate H7N9 vaccine strains using reverse genetics, consisting of hemagglutinin and neuraminidase genes derived from a human H7N9 virus and the remaining genes from the PR8 (A/PuertoRico/8/34 (H1N1)) virus. This H7N9 vaccine exhibited superior efficacy when combined with MF59 compared to other adjuvants. Immunized BALB/c mice were followed to determine the duration of the protective immune response. Antibody levels decreased to between one-half and one-eighth of the peak values four months after the final dose of the vaccine. Previously vaccinated mice received an A/Zhejiang/DTID-ZJU01/2013 H7N9 challenge six months post-vaccination, and all remained protected. We also verified that MF59 enhanced the HI, MN, and IgG antibody titers to influenza antigens. The humoral immune response and Th2 cytokine production following influenza challenge was potently induced in the animals that received the split vaccine. Therefore, the split H7N9 influenza vaccine with the MF59 adjuvant could effectively induce antibody production and protect mice from H7N9 virus challenge even after six months.

Open-label phase I clinical trial of Ad5-EBOV in Africans in China.

BACKGROUND: To determine the safety and immunogenicity of a novel recombinant adenovirus type 5 vector based Ebola virus disease vaccine (Ad5-EBOV) in Africans in China. METHODS: A phase 1, dose-escalation, open-label trial was conducted. 61 healthy Africans were sequentially enrolled, with 31 participants receiving one shot intramuscular injection and 30 participants receiving a double-shot regimen. Primary and secondary end points related to safety and immunogenicity were assessed within 28 d after vaccination. This study was registered with ClinicalTrials.gov (NCT02401373). RESULTS: Ad5-EBOV is well tolerated and no adverse reaction of grade 3 or above was observed. 53 (86.89%) participants reported at least one adverse reaction within 28 d of vaccination. The most common reaction was fever and the mild pain at injection site, and there were no significant difference between these 2 groups. Ebola glycoprotein-specific antibodies appeared in all 61 participants and antibodies titers peaked after 28 d of vaccination. The geometric mean titres (GMTs) were similar between these 2 groups (1919.01 vs 1684.70 P = 0.5562). The glycoprotein-specific T-cell responses rapidly peaked after 14 d of vaccination and then decreased, however, the percentage of subjects with responses were much higher in the high-dose group (60.00% vs 9.68%, P = 0.0014). Pre-existing Ad5 neutralizing antibodies could significantly dampen the specific humoral immune response and cellular response to the vaccine. CONCLUSION: The application of Ad5-EBOV demonstrated safe in Africans in China and a specific GP antibody and T-cell response could occur 14 d after the first immunization. This acceptable safety profile provides a reliable basis to proceed with trials in Africa.

Analysis of the immunogenicity and bioactivities of a split influenza A/H7N9 vaccine mixed with MF59 adjuvant in BALB/c mice.

The H7N9 influenza virus caused significant mortality and morbidity in humans during an outbreak in China in 2013. A recombinant H7N9 influenza seed with hemagglutinin (HA) and neuraminidase (NA) gene segments from A/Zhejiang/DTID-ZJU01/2013(H7N9) and six internal protein gene segments from A/Puerto Rico/8/34(H1N1; PR8) were generated using reverse genetics. We sought to determine the immunogenic, protective properties, and mechanisms of a split avian influenza A/H7N9 vaccine mixed with MF59 adjuvant in comparison to vaccines that included other adjuvant. BALB/c mice were vaccinated with two doses of different amounts and combinations of this novel A/ZJU01/PR8/2013 split vaccine with adjuvant. Mice were subsequently challenged with A/Zhejiang/DTID-ZJU01/2013(H7N9) by intranasal inoculation. We verified that MF59 enhanced the HI, MN, and IgG antibody titers to influenza antigens. Compared with alum, MF59 could more potentially induce humoral immune responses and Th2 cytokine production after virus infection, while both MF59 and alum can slightly increase NK cell activity. This split H7N9 influenza vaccine with MF59 adjuvant could effectively induce antibody production and protect mice from H7N9 virus challenge. We have selected this vaccine for manufacture and future clinical studies to protect humans from H7N9 virus infection.

Preclinical evaluation of the safety and pathogenicity of a live attenuated recombinant influenza A/H7N9 seed strain and corresponding MF59-adjuvanted split vaccine.

Developing a safe and effective H7N9 influenza vaccine was initiated in early spring 2013, following human infections with a novel avian influenza A (H7N9) virus. In this study, a candidate H7N9 vaccine seed strain is produced using reverse genetics, with HA and NA derived from a human H7N9 virus and the remaining genes from the PR8 backbone virus which grows well in eggs. We verified that the virulence and transmissibility of the recombinant H7N9 vaccine seed strain were decreased as compared to wild-type H7N9 virus, to levels comparable with PR8. Using the seed virus, we produced a monovalent split influenza A (H7N9) MF59-adjuvanted vaccine that was immunogenic in mice. Our H7N9 vaccine is selected for clinical investigation and potential human use. To assess the safety of our H7N9 vaccine, we performed acute toxicity, repeated dose toxicity and active systemic anaphylaxis tests. Our results showed that, under the conditions used in this study, the NOEAL (no obvious adverse effect level) was 30 µg/0.5 mL.

Successful treatment of avian-origin influenza A (H7N9) infection using convalescent plasma.

In January 2015, there was an outbreak of avian-origin influenza A (H7N9) virus in Zhejiang Province, China. A 45-year-old man was admitted to the First Affiliated Hospital of Zhejiang University with a high fever that had lasted 7 days, chills, and a cough with yellow sputum. Laboratory testing confirmed infection with the H7N9 virus, likely obtained from contact with poultry at a local live poultry market. A large dense shadow was apparent in the patient's left lung at the time of admission. Treatment with oseltamivir (75mg twice daily) did not improve the patient's condition. The decision was made to try using convalescent plasma to treat the infection. Convalescent plasma was administered 3 days after the patient was admitted to the hospital and led to a marked improvement. To our knowledge, this is the first report of the successful use of convalescent plasma to treat a case of H7N9 infection in China. These results suggest that the combination of convalescent plasma and antiviral drugs may be effective for the treatment of avian-origin H7N9 infection.

Surgical treatment for the proliferative retinal detachment associated with macular hole in the morning glory syndrome.

PURPOSE: To evaluate the efficacy of vitrectomy with peripapillary photocoagulation and silicone oil tamponade for the proliferative retinal detachment associated with macular hole in children with morning glory syndrome. METHODS: Eight children with morning glory syndrome (mean age 8.0 +/- 2.8 years; range 5-13 years) were included; all patients had unilateral eye disease and were initially misdiagnosed as having bilateral squint or amblyopia, with best corrected visual acuity < 6/60. Five patients could not cooperate with the fundus examination and one patient had lens opacities. B-ultrasound confirmed that all eight patients had retinal detachment and optic disc dysplasia. All patients underwent standard 3-port pars plana vitrectomy surgery (20G for three cases and 23G for five cases). At surgery, all patients were confirmed to have morning glory syndrome, macular hole, and proliferative retinal detachment; two cases had a funnel-shaped bulge. All the retinal detachments involved the macular area, and macular hole was detected in the abnormal expansion excavation of the optic disk. The epiretinal membrane and subretinal membrane were completely removed during surgery. Combined photocoagulation in the abnormal expansion excavation of the optic disk, and silicone oil tamponade were also performed. RESULTS: All eyes achieved anatomical resolution of retinal detachment. After follow-ups ranging from eight months to four years, the visual function for all patients was improved by postoperative refractive correction associated with vision training. Best corrected visual acuity was 6/600 to 6/30 at the final follow-up, no retinal detachment recurred, and no silicone oil fluid entered the subretinal space. The silicone oil was successfully removed postoperatively after a mean of 1.5 years. CONCLUSION: Vitrectomy with peripapillary photocoagulation and silicone oil tamponade is effective in treating the proliferative retinal detachment associated with macular hole in children with morning glory syndrome.