[Analysis of the correlation between fatty liver index and myocardial remodeling].

Objective: To analyze the correlation between fatty liver index (FLI) and myocardial remodeling. Methods: For cross-sectional study, cluster sampling was used to conduct a follow-up study of "Risk evaluation of cancers in Chinese diabetic individuals: A longitudinal (REACTION) study" among communities of Gucheng and Pingguoyuan of Beijing from April 2015 to September 2015. According to the inclusion and exclusion criteria, 8 848 participants were selected. Biochemical indicators such as body mass index, waist circumference, triglycerides, and γ-glutamyl transpeptidase were detected to calculate the FLI. The correlation between FLI and myocardial remodeling was analyzed. Interventricular septal thickness (IVS), left atrial diameter (LAD), left ventricular end diastolic diameter (LVEDD), and the presence of diastolic dysfunction were measured by color doppler ultrasound. The participants were divided into Q1 group (FLI<30, 4 529 cases), Q2 group (30≤FLI<60, 2 762 cases), and Q3 group (FLI≥60, 1 557 cases) based on FLI levels. Single factor analysis of variance was used for inter-group comparison, logistic regression analysis was used to analyze the correlation between FLI and myocardial remodeling. Results: A total of 8 848 subjects were selected for the study (3 110 male and 5 738 female, mean age: 59.96 years). The IVS of Q1, Q2, and Q3 groups were (9.35±1.08), (9.73±1.22), and (10.07±1.31) mm, respectively. The LAD were (30.94±3.90), (33.37±4.12), and (34.98±4.47) mm, respectively. The LVEDD were (42.51±5.05), (44.43±5.10), and (46.06±5.52) mm, respectively. All increased with the increase of FLI (all P<0.001). FLI was an independent risk factor for IVS thickening, LAD increase, LVEDD increase, and diastolic function decrease. The respective risks for IVS thickening, LAD increase, LVEDD increase, and diastolic function decrease in a population with intermediate and higher FLI levels was 1.62 times (95%CI 1.39-1.89) and 2.53 times (95%CI 2.13-3.00); 2.71 times (95%CI 2.39-3.06) and 5.00 times (95%CI 4.12-6.08); 2.36 times (95%CI 1.85-3.00) and 4.33 times (95%CI 3.33-5.62); and 1.90 times (95%CI 1.63-2.19) and 1.95 times (95%CI 1.60-2.37) than those with lower FLI levels. Conclusion: There is a certain relevance between FLI and myocardial remodeling.

[Study on the relationship between hemoglobin glycosylation index and arteriosclerosis- related blood lipids].

Objective: To study the relationship between hemoglobin glycation index (HGI) and blood lipid indices such as low-density lipoprotein cholesterol (LDL-C), non-high-density lipoprotein cholesterol (non-HDL-C), and plasma atherogenic index (AIP). Methods: This cross-sectional study included 16 049 participants from the Beijing Apple Garden community between December 2011 and August 2012. The subjects were divided into three groups based on the HGI quartile: low (n=5 388), medium (n=5 249), and high (n=5 412). The differences in blood lipid indicators between different HGI groups were compared and multivariate logistic regression model was established to analyze the association between HGI and dyslipidemia. And multivariate logistic regression model was established to analyze the relationship between HGI and blood lipid indicators in different glucose metabolism populations. Results: There were 16 049 participants in all (mean age: 56 years), including 10 452 women (65.1%). They were classified into normal glucose tolerance (9 093 cases), prediabetes (4 524 cases), and diabetes (2 432 cases) based on glucose tolerance status. In the general population, with the increase of HGI, LDL-C, non-HDL-C, and AIP gradually increased (all P values for trends were <0.05), and the proportion of abnormalities increased significantly (χ2=101.40, 42.91, 39.80; all P<0.001). A multivariate logistic regression model was established, which suggested a significant correlation between HGI and LDL-C, non-HDL-C, and AIP (all P<0.05), after adjusting for factors such as age, sex, fasting blood glucose, hypertension, body mass index, smoking, and alcohol consumption. In the overall population, normal glucose tolerance group, and diabetes group, HGI had the highest correlation with non-HDL-C (OR values of 1.325, 1.678, and 1.274, respectively); in the prediabetes group, HGI had a higher correlation with LDL-C (OR value: 1.510); and in different glucose metabolism groups, AIP and HGI were both correlated (OR: 1.208-1.250), but not superior to non-HDL-C and LDL-C. Conclusion: HGI was closely related to LDL-C, non HDL-C, and AIP in the entire population and people with different glucose metabolism, suggesting that HGI may be a predictor of atherosclerotic cardiovascular disease.

[Study on nomograph predicting the risk of type 2 diabetes mellitus in Beijing community adults].

Objective: Development and validation of a nomogram for predicting the 4-year incidence of type-2 diabetes mellitus (T2DM) in a Chinese population was attempted. Methods: This prospective cohort study was conducted in Shijingshan District Pingguoyuan Community (Beijing, China) from December 2011 to April 2012 among adults aged≥40 years not suffering from T2DM. Finally, 8 058 adults free of T2DM were included with a median duration of follow-up of 4 years. Participants were divided into a modeling group and verification group using simple random sampling at a ratio of 7∶3. Univariate and multivariate Cox proportional risk models were applied to identify the independent risk predictors in the modeling group. A nomogram was constructed to predict the 4-year incidence of T2DM based on the results of multivariate analysis. The Concordance Index and calibration plots were used to evaluate the differentiation and calibration of the nomogram in both groups. Results: A total of 5 641 individuals were in the modeling group and 2 417 people were in the validation group, of which 265 and 106 had T2DM, respectively, at 4-year follow-up. In the modeling group, age (HR=1.349, 95%CI 1.011-1.800), body mass index (HR=1.347, 95%CI 1.038-1.746), hyperlipidemia (HR=1.504, 95%CI 1.133-1.996), fasting blood glucose (HR=4.189, 95%CI 3.010-5.830), 2-h blood glucose level according to the oral glucose tolerance test (HR=3.005, 95%CI 2.129-4.241), level of glycosylated hemoglobin (HR=3.162, 95%CI 2.283-4.380), and level of γ-glutamyl transferase (HR=1.920, 95%CI 1.385-2.661) were independent risk factors for T2DM. Validation of the nomogram revealed the Concordance Index of the modeling group and validation group to be 0.906 (95%CI 0.888-0.925) and 0.844 (95%CI 0.796-0.892), respectively. Calibration plots showed good calibration in both groups. Conclusion: These data suggest that our nomogram could be a simple and reliable tool for predicting the 4-year risk of developing T2DM in a high-risk Chinese population.

[Predictive value of hemoglobin glycation index for chronic kidney disease].

Objective: To investigate the influence of hemoglobin glycation index (HGI) on the risk of incident chronic kidney disease (CDK) among nondiabetic patients. Methods: Prospective cohort study. At baseline, a total of 7 407 nondiabetic patients without a history of CKD from Pingguoyuan Community of the Shijingshan District in Beijing were included from December 2011 to August 2012, who were then divided into three groups according to the tertiles of their baseline HGI levels. The CKD incidence rate was compared among the different HGI groups at last follow-up. Cox multivariable regression was applied to evaluate whether HGI measures predicted CKD risk. Test for trend across tertiles were examined using ordinal values in separate models. Results: The mean age of the subjects was (56.4±7.5) years, and 4 933 (66.6%) were female. At mean follow-up of 3.23 years, 107 (1.4%) individuals developed CKD. The incidence of CKD was gradually increasing from the low to high HGI groups [1.1% (28/2 473) vs. 1.2% (31/2 564) vs. 2.0% (48/2 370), P=0.016]. In the multivariate Cox regression analysis, after adjustment for potential confounders, the high HGI group had a 68.5% increased risk of CKD compared with the low HGI group (HR=1.685, 95%CI 1.023 to 2.774). CKD risk increased with increasing HGI tertiles (P for trend=0.028). Conclusion: High HGI is associated with an increased risk for CKD in the nondiabetic population, indicating that HGI may help identify individuals at high risk for CKD.

[Research progress of induced pluripotent stem cells in promoting wound healing of diabetic foot ulcers].

Chronic wounds such as diabetic foot ulcers are epidemic, which bring huge burdens to both the patients and the society. However, with current treatment methods, diabetic foot ulcers often heal poorly and recur frequently, so it is urgent and important to find new and advanced therapies. Stem cell therapy has been proved by a large number of pre-clinical and clinical studies as a potential treatment for chronic wounds. However, the acquisition of stem cells often depends on invasive techniques, and immunogenicity and limited cell survival in vivo also limit the large-scale application and promotion of stem cell therapy. In the recent years, with the development and advance of induced pluripotent stem cell (iPSC) technology, it has shown a strong translational potential in the treatment of chronic wounds such as diabetic foot ulcers. This article reviews the applications and prospect of iPSCs in animal wound healing models including diabetic ulcers and limb ischemia, the limitations of their clinical application, and the methods to improve their safety.

Aetiology of tinea capitis in China: a multicentre prospective study.

BACKGROUND: Tinea capitis is still common in developing countries, such as China. Its pathogen spectrum varies across regions and changes over time. OBJECTIVES: This study aimed to clarify the current epidemiological characteristics and pathogen spectrum of tinea capitis in China. METHODS: A multicentre, prospective descriptive study involving 29 tertiary hospitals in China was conducted. From August 2019 to July 2020, 611 patients with tinea capitis were enrolled. Data concerning demography, risk factors and fungal tests were collected. When necessary, the pathogens were further identified by morphology or molecular sequencing in the central laboratory. RESULTS: Among all enrolled patients, 74·1% of the cases were in patients aged 2-8 years. The children with tinea capitis were mainly boys (56·2%) and were more likely than adults to have a history of animal contact (57·4% vs. 35·3%, P = 0·012) and zoophilic dermatophyte infection (73·5% vs. 47%). The adults were mainly female (83%) and were more likely than children to have anthropophilic agent infection (53% vs. 23·9%). The most common pathogen was zoophilic Microsporum canis (354, 65·2%), followed by anthropophilic Trichophyton violaceum (74, 13·6%). In contrast to the eastern, western and northeastern regions, where zoophilic M. canis predominated, anthropophilic T. violaceum predominated in central China (69%, P < 0·001), where the patients had the most tinea at other sites (20%) and dermatophytosis contact (26%) but the least animal contact (39%). Microsporum ferrugineum was the most common anthropophilic agent in the western area, especially in Xinjiang province. CONCLUSIONS: Boys aged approximately 5 years were the most commonly affected group. Dermatologists are advised to pay more attention to the different transmission routes and pathogen spectra in different age groups from different regions.

Isolation and characterization of a goose astrovirus 1 strain causing fatal gout in goslings, China.

In recent years, goose gout, a severe infectious disease, has affected the development of the goose industry in China. Two different genotypes of goose astrovirus (GAstV), named as GAstV-1 and GAstV-2, were identified. GAstV-2 viruses are known to be the causative agent of goose gout; however, GAstV-1 has not been isolated, and the relationship between GAstV-1 and goose gout is unknown. One full genome sequence, designated as GAstV/CHN/TZ03/2019 (TZ03), was determined from the clinical tissue samples of a diseased gosling using next-generation sequencing. The complete genome of TZ03 was 7,262 nucleotides in length with typical genomic characteristics of avastroviruses. The TZ03 strain shares the highest identity (96.6%) with the GAstV-1 strain FLX, but only 51.5 to 61.3% identity with other astroviruses in Avastrovirus. Phylogenetic analysis revealed that the TZ03 strain clustered together with the GAstV-1 strains FLX and AHDY and was highly divergent from GAstV-2 viruses. The TZ03 strain was successfully isolated from goose embryos and caused 100% mortality of goose embryos after 5 passages. Electron microscopy showed that the virus particles were spherical with a diameter of ∼22 nm. The clinical symptoms were reproduced by experimental infection of healthy goslings, which were similar to those caused by GAstV-2 strains. Our data show that GAstV-1 is one of the causative agents of the ongoing goose gout disease in China. These findings enrich our understanding of the evolution of GAstVs that cause gout and provide potential options for developing biological products to treat goose gout.

[Influence of maggot excretions/secretions on the anti-Pseudomonas aeruginosa effect of neutrophils in patients with diabetic foot ulcer].

Objective: To investigate the effects of medical maggot excretions/secretions (ES) on neutrophils phagocytosis and bactericidal effect in patients with diabetic foot ulcer (DFU). Methods: The experimental research method was used. Thirty DFU patients (16 males and 14 females, aged (64±7) years)who were admitted to the Diabetes Foot Center, the Department of Endocrinology of Air Force Hospital of Eastern Theater Command from June to December 2020 and met the inclusion criteria were recruited. Discontinuous percoll gradient centrifugation method was used to separate the neutrophils. Cells from each patient were enrolled into normal saline group and maggot ES group (30 wells in each group), respectively; sterile normal saline and ES with a final mass concentration of 357 µg/mL (the same as below) were added, respectively. After 1 and 2 hour(s) of culture, the phagocytosis rate and phagocytic index of cells were observed and counted under Wright's staining. Ten patients were selected, then the cells of each patient were enrolled into Pseudomonas aeruginosa+neutrophils group and Pseudomonas aeruginosa+neutrophils+maggot ES group (10 wells in each group) and were treated corresponding, respectively. Pseudomonas aeruginosa alone group and Pseudomonas aeruginosa+maggot ES group (10 wells in each group) were set up respectively; Pseudomonas aeruginosa+RPMI 1640 culture medium+sterile normal saline and Pseudomonas aeruginosa+RPMI 1640 culture medium+maggot ES were added, respectively. After 2 hours of culture, the number of viable bacteria colony was counted by plate colony number method. Six, six, and three patients were selected respectively, and the cells of each patient were respectively enrolled into maggot ES group and normal saline group (6, 6, and 3 wells in each group, respectively) and treated accordingly. After 6 hours of culture, real-time fluorescent quantitative reverse transcription polymerase chain reaction was used to detect the mRNA expressions of interleukin 1ß (IL-1ß), IL-6, and lysozyme in cells, the content of IL-1ß and IL-6 in cell culture supernatant were determined by enzyme-linked immunosorbent assay, and the positive cells expressing lysozyme were observed with immunofluorescence method. Data were statistically analyzed with one-way analysis of variance, paired sample t test, least significant difference test, and Wilcoxon rank sum test. Results: After 1 hour of culture, the phagocytosis rate and phagocytic index of cells in maggot ES group (53.5% (49.7%, 58.0%) and 3.18 (2.96, 3.32)) were similar to 52.0% (47.5%, 55.2%) and 3.15 (2.96, 3.25) of normal saline group (Z=-1.701, -1.092, P>0.05). After 2 hours of culture, the phagocytosis rate and phagocytic index of cells in maggot ES group (70.0% (66.7%, 72.0%) and 4.47 (4.22, 4.96)) were significantly higher than 58.0% (55.0%, 60.0%) and 4.11 (3.52, 4.24) in normal saline group (Z=-4.786, -4.279, P<0.01). After 2 hours of culture, the number of viable bacteria colony in Pseudomonas aeruginosa+neutrophils group was significantly lower than that in Pseudomonas aeruginosa alone group (P<0.01), and the number of viable bacteria colony in Pseudomonas aeruginosa+neutrophils+maggot ES group was significantly lower than that in Pseudomonas aeruginosa+maggot ES group and Pseudomonas aeruginosa+neutrophils group (P<0.01). After 6 hours of culture, the mRNA expressions of IL-1ß, IL-6, and lysozyme of cells in maggot ES group were significantly higher those in normal saline group (t=-3.279, -4.273, -4.763, P<0.05 or P<0.01); the concent of IL-1ß and IL-6 in cell culture supernatant of maggot ES group were significantly higher than those of normal saline group (t=-9.526, -6.447, P<0.01); there were significantly more positive cells expressing lysozyme in maggot ES group than in normal saline group. Conclusions: Maggot ES can enhance the phagocytosis and bactericidal effect of neutrophils on Pseudomonas aeruginosa by promoting the production of neutrophils immune defense related cytokines and lysozyme in DFU patients.

[Mechanism of maggot debridement therapy in promoting wound angiogenesis in patients with diabetic foot ulcer].

Objective: To investigate the mechanism of maggot debridement therapy (MDT) in promoting wound angiogenesis in patients with diabetic foot ulcer (DFU). Methods: (1) From June 2018 to June 2019, the patients admitted to Nanjing Junxie Hospital who met the inclusion criteria were recruited, including 12 DFU patients given MDT for three days [6 males and 6 females, aged (56±12) years] and 12 acute trauma patients without diabetes mellitus [6 males and 6 females, aged (53±10) years], who were enrolled into DFU group and non-diabetic trauma group respectively. Before and after application of MDT, the wound characteristics of patients in DFU group were observed and the wound tissue samples were taken. The wound tissue in non-diabetic trauma group was taken at patient's first visit before debridement. The expression of angiogenesis marker CD31 in the wound tissue of patients in DFU group was detected by immunohistochemistry before and after application of MDT. Western blotting and real-time fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR) were used respectively to detect the protein and mRNA expressions of fatty acid synthase (FAS) in wound tissue of patients in DFU group before and after application of MDT and in non-diabetic trauma group before debridement. (2) Human umbilical vein endothelial cells (HUVECs) were cultured in endothelial cell culture medium containing 10% fetal bovine serum. The 3rd to 6th passages of cells in logarithmic growth phase were used in the following experiments. Excretions/secretions (ES) were extracted from 3-day-old sterile Lucilia sericata larvae for subsequent experiments. Three batches of cells were divided into phosphate buffer solution (PBS) control group, high glucose alone group, high glucose+ 5 µg/mL maggot ES group, and high glucose+ 10 µg/mL maggot ES group, which were treated with PBS, glucose in final molarity concentration of 20 mmol/L, glucose in final molarity concentration of 20 mmol/L+ maggot ES in final mass concentration of 5 µg/mL, and glucose in final molarity concentration of 20 mmol/L+ maggot ES in final mass concentration of 10 µg/mL respectively. The total volume of reagents in each group was the same. After 48 hours of culture, Western blotting, real-time fluorescent quantitative RT-PCR and immunofluorescence method were used to detect the protein and mRNA expressions of FAS in each batch of cells and the expression and localization of FAS protein in cells respectively. The number of samples for mRNA expression was 3. (3) Two batches of cells were divided into small interference RNA (siRNA) alone group, siRNA control+ maggot ES group and siRNA-FAS+ maggot ES group, which were transfected with 100 µmol/L (final molarity concentration) insignificant control siRNA, insignificant control siRNA, and siRNA-FAS for 4-6 h respectively, and then they were routinely cultured for 24 h with PBS added, maggot ES in final mass concentration of 10 µg/mL, and maggot ES in final mass concentration of 10 µg/mL respectively. The total volume of reagents in each group was the same. One batch of cells was used for scratch test, the scratch width was observed at 24 hour after scratching to detect the cell migration ability; one batch of cells was subjected to tube forming experiment, and the formation of cell tubules was observed after 24 hours of culture. The number of samples was 3 in scratch test and tube forming experiments. Data were statistically analyzed with t test, one-way analysis of variance, least significant difference test, analysis of variance for repeated measurement, and Bonferroni method. Results: (1) Compared with those before application of MDT, fresh granulation tissue significantly increased and necrotic tissue decreased obviously in wound, and the expression of CD31 significantly increased in wound tissue of patients in DFU group after application of MDT. The expression of FAS protein in wound tissue of patients in DFU group before application of MDT was significantly lower than that in non-diabetic trauma group before debridement, and the expression of FAS protein in wound tissue of patients in DFU group after application of MDT was significantly higher than that before application of MDT. The expression of FAS mRNA in wound tissue of patients in DFU group before application of MDT was 1.00±0.17, which was significantly less than 3.87±1.02 in non-diabetic trauma group before debridement (t=9.808, P<0.01). The expression of FAS mRNA in wound tissue of patients in DFU group after application of MDT was 1.85±0.31, which was significantly higher than that before application of MDT (t=-10.853, P<0.01). (2) After 48 hours of culture, Western blotting detection showed that the expression of FAS protein in cells in high glucose alone group was significantly less than that in PBS control group, and the expressions of FAS protein in cells in high glucose+ 5 µg/mL maggot ES group and high glucose+ 10 µg/mL maggot ES group were significantly higher than the expression in high glucose alone group. Real-time fluorescent quantitative RT-PCR determination showed that the expression of FAS mRNA in cells in high glucose alone group was 0.392±0.073, which was significantly lower than 1.000±0.085 in PBS control group (P<0.01); there was statistically significant difference between the expression of FAS mRNA in cells in high glucose+ 5 µg/mL maggot ES group (0.561±0.047) and that in high glucose+ 10 µg/mL maggot ES group (0.687±0.013) (P<0.05), both of which were significantly higher than the expression in high glucose alone group (P<0.01). The results of immunofluorescence detection showed that FAS protein was mainly located in the cytoplasm of cells in each group, and its expression was similar to that detected by Western blotting. (3) At 24 hour after scratch, the uncured widths of cell scratch in siRNA control+ maggot ES group and siRNA-FAS+ maggot ES group were significantly narrower than the uncured width in siRNA alone control group (P<0.01), and the uncured width of cell scratch in siRNA-FAS+ maggot ES group was significantly wider than that in siRNA control+ maggot ES group (P<0.01). After 24 hours of culture, the numbers of tubules in siRNA+ maggot ES group and siRNA-FAS+ maggot ES group were significantly more than the number in siRNA alone control group (P<0.05 or P<0.01), and the number of tubules in siRNA-FAS+ maggot ES group was obviously less than that in siRNA control+ maggot ES group (P<0.05). Conclusions: MDT up-regulates the expression of FAS through maggot ES, which promotes the activity of vascular endothelial cells, thus promoting the wound angiogenesis in patients with DFU.

Coexistence of hyper-uricaemia and low urinary uric acid excretion further increases risk of chronic kidney disease in type 2 diabetes.

AIM: To investigate whether hyper-uricaemia and decreased urinary uric acid excretion (UUAE) are associated with increased risk of chronic kidney disease (CKD), and whether the coexistence of hyper-uricaemia and low UUAE further increases CKD risk in type 2 diabetes mellitus (T2DM). METHODS: In this cross-sectional study based on serum uric acid (SUA) and UUAE levels, 2846 T2DM inpatients were divided into those with normal SUA and UUAE (group 1), normal SUA and low UUAE (group 2), hyper-uricaemia and normal UUAE (group 3), and hyper-uricaemia and low UUAE (group 4). Hyper-uricaemia was defined as SUA levels ≥ 420 µmol/L in men and ≥ 360 µmol/L in women. Low UUAE was defined as levels below the first UUAE quintiles (< 2161 µmol/24 h in men, 1977 µmol/24 h in women). RESULTS: There were trends for significantly increased prevalences of CKD (4.3%, 12.6%, 18.3%, 47.8%; P < 0.001), albuminuria (20.2%, 26.4%, 36.9%, 54.9%; P < 0.001) and macroalbuminuria (3.3%, 10.1%, 10.7%, 31.9%; P < 0.001) from groups 1 to 4, respectively. After controlling for multiple confounding factors, prevalences of CKD (P < 0.001) and urinary albumin levels (P = 0.013) showed significantly increasing trends, whereas eGFR levels were markedly decreased from groups 1 to 4 (P < 0.001). CONCLUSION: Hyper-uricaemia and low UUAE levels are closely associated with presence of CKD, and the concomitant presence of hyper-uricaemia and decreased UUAE levels further increased CKD risk in T2DM. Thus, the combined consideration of SUA and UUAE levels may help to identify those T2DM patients at higher CKD risk.

Expression of duck hepatitis A virus type 1 VP3 protein mediated by avian adeno-associated virus and its immunogenicity in ducklings.

The avian adeno-associated virus (AAAV) is a replication-defective nonpathogenic virus that has been proved to be useful as a viral vector in gene delivery. In this study, the feasibility of AAAV for transgenic expression of duck hepatitis A virus (DHAV) VP3 structural protein and its ability to induce protective immunity in ducklings was assessed. The recombinant AAAV (rAAAV-VP3) expressing the VP3 protein was prepared by co-infection of Sf9 cells with recombinant baculovirus (rBac-VP3) containing VP3 gene flanked by inverted terminal repeats (ITRs) of AAAV and the other two recombinant baculovirus expressing AAAV functional and structural genes, respectively. The generation of rAAAV-VP3 was demonstrated by electron microscopy, immunofluorescence assay, and western blot analysis. One day old ducklings were inoculated with rAAAV-VP3 or commercial attenuated vaccine and then challenged with DHAV-1 strain SH two weeks post vaccination. Anti-DHAV-1 antibodies were detected in all vaccinated groups by ELISA, and the titers between the rAAAV-VP3 group and the attenuated vaccine group were not statistically significant. Real time RT-PCR analysis showed that the virus copy numbers in the livers of the PBS control group were significantly higher than that of the rAAAV-VP3 and attenuated vaccine groups. In conclusion, we demonstrated that the VP3 expression mediated by rAAAV in ducklings could induce protective immunity against DHAV challenge, and this could be a candidate vaccine for the control of duck viral hepatitis. Keywords: avian adeno-associated virus; duck hepatitis A virus; VP3 gene; immunogenicity.

Protection against duck hepatitis a virus type 1 conferred by a recombinant avian adeno-associated virus.

The avian adeno-associated virus (AAAV) has been proved to be an efficient gene transfer vector for human gene therapy and vaccine research. In this experiment, an AAAV-based vaccine was evaluated for the development of a vaccine against duck hepatitis a virus type 1 (DHAV-1). The major capsid VP1 gene was amplified and subcloned into pFBGFP containing the inverted terminal repeats of AAAV, and then the recombinant baculovirus rBac-VP1 was generated. The recombinant AAAV expressing the VP1 protein (rAAAV-VP1) was produced by co-infecting Sf9 cells with rBac-VP1 and the other 2 baculoviruses containing AAAV functional genes and structural genes respectively, and confirmed by electron microscopy, Western blotting and immunofluorescence assays. Quantitative real-time PCR revealed that the titer of rAAAV-VP1 was about 9 × 1012 VG/mL. Immunogenicity was studied in ducklings. One day ducklings were injected intramuscularly once with rAAAV-VP1. Serum from rAAAV-VP1-vaccinated ducklings showed a systemic immune response evidenced by VP1-specific enzyme-linked immunosorbent assay and virus neutralization test. Furthermore, all ducklings inoculated with rAAAV-VP1 were protected against DHAV-1 challenge. The data of quantitative real-time RT-PCR from livers of challenged ducklings also showed that the level of virus copies in rAAAV-VP1 group was significantly lower than that of the PBS group. Collectively, these results demonstrate that the AAAV-based vaccine is a potential vaccine candidate for the control of duck viral hepatitis.

Prevalence and clinical characteristics of non-alcoholic fatty liver disease in newly diagnosed patients with ketosis-onset diabetes.

AIM: As the prevalence and clinical characteristics of non-alcoholic fatty liver disease (NAFLD) are still unknown in ketosis-onset diabetes, the present study compared the characteristics of NAFLD in type 1 diabetes (T1D), ketosis-onset and non-ketotic type 2 diabetes (T2D) patients. METHODS: This cross-sectional study was performed with newly diagnosed Chinese patients with diabetes, including 39 T1D, 165 ketosis-onset and 173 non-ketotic T2D, with 30 non-diabetics included as controls. NAFLD was determined by hepatic ultrasonography, then its clinical features were analyzed and its associated risk factors evaluated. RESULTS: NAFLD prevalence in patients with ketosis-onset diabetes (61.8%) was significantly higher than in controls (23.3%; P=0.003) and in T1D patients (15.4%; P<0.001). However, there was no difference in prevalence between ketosis-onset and non-ketotic T2D patients (52.6%; P=0.229), although BMI and alanine aminotransferase (ALT) proved to be independent risk factors for the presence of NAFLD in both these groups whereas, in T1D patients, serum uric acid levels were independent risk factors. CONCLUSION: NAFLD prevalence and risk factors in ketosis-onset diabetes were similar to those in non-ketotic T2D, but different from those in T1D. These data provide further evidence that ketosis-onset diabetes should be classified as a subtype of T2D rather than idiopathic T1D.

[Evaluation of transperineal sonography for lower urinary tract symptoms after pelvic floor reconstruction].

Objective: To evaluate transperineal sonography for lower urinary tract symptoms after pelvic floor reconstruction. Methods: Eighty-three patients with severe pelvic organ prolapse received surgeries in Fuzhou General Hospital from September 2014 to September 2015, dividing into two groups: 27 patients were selected to receive transvaginal mesh (TVM) pelvic floor reconstruction surgery with tension-free vaginal tape-Abbrevo (TVT-Abbrevo) incontinence surgery, named TVM+TVT-Abbrevo group; 56 patients were selected to receive TVM pelvic floor reconstruction surgery only, named TVM group. The ultrasonic parameters at rest, on contraction and Valsalva condition respectively were observed and measured, including the bladder neck descent (BND), urethral rotation angle, retrovesical angle, levator urethra gap (LUG), the existence of bladder neck funneling, position of the tape, by using 2D and 3D transperineal ultrasound. Results: The two groups were compared with the ultrasonic parameters before and after operation: two groups of patients with postoperative BND [(2.3±0.5) versus (3.1±0.7) cm, (1.6±0.4) versus (3.6±0.4) cm] were significantly reduced, the difference was statistically significant (P=0.02, P<0.01). The two groups of LUG before and after operation [(3.62±0.45) versus (3.26±0.92) cm, (2.96±0.47) versus (2.72±0.38) cm] both had significant difference by maximum Valsalva (P<0.01, P=0.04). There was statistical significance difference of urethral rotation angle in TVM+TVT-Abbrevo group by maximum Valsalva (P=0.01). Observation of morphology: (1) 2 patients with difficulty in urination in TVM+TVT-Abbrevo group, ultrasound showed when the position of the bladder down the urethra discount; 4 patients with stress urinary incontinence (SUI), ultrasound showed slings off or release. (2) One patient with difficulty in urination in TVM group, but ultrasound showed lower urinary tract anatomy were normal; 5 patients with SUI, ultrasound showed the position of the bladder neck were significantly lower in 3 patients, showing high mobility, and the other 2 patients had a larger urethral diameter, showing a tendency of natural deletion. Conclusion: s Anatomy of lower urinary tract could be clearly showed by transperineal sonography. This could provide imaging support for the diagnosis of lower urinary tract symptoms after pelvic floor reconstruction.

Study on the expression of human lysozyme in oviduct bioreactor mediated by recombinant avian adeno-associated virus.

Due to its antimicrobial properties and low toxicity, human lysozyme (hLYZ) has broad application in the medical field and as a preservative used by the food industry. However, limited availability hinders its widespread use. Hence, we constructed a recombinant avian adeno-associated virus (rAAAV) that would specifically express hLYZ in the chicken oviduct and harvested hLYZ from the egg whites of laying hens. The oviduct-specific human lysozyme expression cassette flanked by avian adeno-associated virus (AAAV) inverted terminal repeats (ITRs) was subcloned into the modified baculovirus transfer vector pFBX, and then the recombinant baculovirus rBac-ITRLYZ was generated. The recombinant avian adeno-associated virus was produced by co-infecting Sf9 cells with rBac-ITRLYZ and the other 2 baculoviruses containing AAAV functional genes and structural genes, respectively. Electron microscopy and real-time PCR revealed that the recombinant viral particles were generated successfully with a typical AAAV morphology and a high titer. After one intravenous injection of each laying hen with 2 × 1011 viral particles, oviduct-specific expression of recombinant human lysozyme (rhLYZ) was detected by reverse transcription-PCR. The expression level of rhLYZ in the first wk increased to 258 ± 11.5 µg/mL, reached a maximum of 683 ± 16.4 µg/mL at the fifth wk, and then progressively declined during the succeeding 7 wk of the study. Western blotting indicated that the oviduct-expressed rhLYZ had the same molecular weight as the natural enzyme. These results indicate that an efficient and convenient oviduct bioreactor mediated by rAAAV has been established, and it is useful for production of other recombinant proteins.

[The clinical feature and microbiological etiology of patients with infective endocarditis].

Objective: To investigate the clinical features, pathogenic distribution and drug susceptibility of patients with infective endocarditis (IE). Methods: Clinical data of IE patients were collected, who were admitted to Capital Medical University Affiliated Beijing Anzhen Hospital from January 2012 to March 2015. Results: Three hundred and three IE patients were enrolled with age of (43±16) years old. Fever (85.5%)and cardiac murmur (62.4%)were the most common clinical presentations. Congenital heart diseases was the leading underlying diseases in IE patients. Non rheumatic valve diseases (13.5%) followed. Vegetations were found in 90.4%(274/303) patients. Streptococcus which accounted for 44.2% was the major pathogen of IE. Staphylococcus (28.9%)was the second common pathogen. Gram-negative bacteria were diversified in categories. Gram-positive cocci were consistently sensitive to vancomycin and linezolid. Conclusions: Congenital heart disease is the main underlying disease related to IE. Streptococcus is still the primary pathogen. Gram-positive cocci keep good sensitivity to vancomycin and linezolid.

[Value of pudendal nerve stimulation for predicting the stress urinary incontinence following pelvic prolapse surgery].

OBJECTIVE: To seek the predictive value of pudendal nerve function that need preventive anti-incontinence surgery at the same time following pelvic prolapse surgery in severe pelvic organ prolapse (POP) patients. METHODS: Seventy women completed this study from January 2014 to June 2015 in Fuzhou General Hospital of Nanjing Military Command, dividing into four groups: POP with or without coexisting occult stress urinary incontinence (OSUI) in preoperation, women with persistent stress urinary incontinence (SUI) in postoperation, women without SUI in postoperation. The pudendal nerve function in preoperation was measured by using Solar Urodynamic Neuro Module, including pudendal nerve terminal motor latency (PNTML), and amplitude. RESULTS: There were statistical significance on bilateral PNTML between POP coexisting OSUI group and only severe POP group [(2.62±0.23) versus (2.40±0.26) ms in right of PNTML, (2.55± 0.21) versus (2.37 ±0.30) ms in left of PNTML; all P<0.05], but no statistical significance on bilateral amplitude (P>0.05). Compared de novo SUI group with POP group in postoperation, de novo SUI group's right of PNTML was significantly increased [(2.74±0.16) versus (2.47±0.26) ms; P< 0.05]; and the right of PNTML was extending 2.5 standard deviation at least compared with the health's [(2.10±0.20) ms]. CONCLUSIONS: The PNTML of pudendal nerve of POP coexisting OSUI is severe than only severe POP, the velocity of nerve conduction is slowing, and PNTML extension has a predictive value for postoperative urinary incontinence. When the right of PNTML of preoperative POP increased by at least 2.5 standard deviations than health's, the risk of SUI postoperative strongly increased, and a anti-incontinence surgery at the same time following pelvic prolapse surgery should be adviced.

[Reported of 2 cases relapsing polychondritis].

Clinical data of two cases of relapsing polychondritis were reported and the relative literatures were reviewed. The two patients diagnosed relapsing polychondritis after clinical and laboratory examinations.The first case was mild , no larynx and trachea involvement, and her condition improved and controlled after glucocorticoid treatment;The second case which involved larynx and trachea caused breathing difficulty. Two years later he died of lung infection and respiratory failure despite a variety of treatment methods. Relapsing polychondritis is very easily misdiagnosed, early diagnosis and treatment can slow the progression of the disease, for patients with violations of the larynx and trachea should pay close attention to disease progression, tracheotomy or place endotracheal memory alloy stent when necessary.