Rapid kidney function decline and increased risk of heart failure in patients with type 2 diabetes: findings from the ACCORD cohort : Rapid kidney function decline and heart failure in T2D.

BACKGROUND: Impaired kidney function and albuminuria are associated with increased risk of heart failure (HF) in patients with type 2 diabetes (T2D). We investigated whether rapid kidney function decline over time is an additional determinant of increased HF risk in patients with T2D, independent of baseline kidney function, albuminuria, and other HF predictors. METHODS: Included in the study were 7,539 participants in the Action to Control Cardiovascular Risk in Diabetes (ACCORD) study with baseline urinary albumin-to-creatinine ratio (UACR) data, who had completed 4 years of follow-up and had ≥ 3 eGFR measurements during that period (median eGFR/year = 1.9, IQR 1.7-3.2). The association between rapid kidney function decline (eGFR loss ≥ 5 ml/min/1.73 m2/year) and odds of HF hospitalization or HF death during the first 4 years of follow-up was estimated by logistic regression. The improvement in risk discrimination provided by adding rapid kidney function decline to other HF risk factors was evaluated as the increment in the area under the Receiving Operating Characteristics curve (ROC AUC) and integrated discrimination improvement (IDI). RESULTS: Over 4 years of follow-up, 1,573 participants (20.9%) experienced rapid kidney function decline and 255 (3.4%) experienced a HF event. Rapid kidney function decline was associated with a ~ 3.2-fold increase in HF odds (3.23, 95% CI, 2.51-4.16, p < 0.0001), independent of baseline CVD history. This estimate was not attenuated by adjustment for potential confounders, including eGFR and UACR at baseline as well as at censoring (3.74; 95% CI 2.63-5.31). Adding rapid kidney function decline during follow-up to other clinical predictors (WATCH-DM score, eGFR, and UACR at study entry and end of follow-up) improved HF risk classification (ROC AUC = + 0.02, p = 0.027; relative IDI = + 38%, p < 0.0001). CONCLUSIONS: In patients with T2D, rapid kidney function decline is associated with a marked increase in HF risk, independent of starting kidney function and/or albuminuria. These findings highlight the importance of serial eGFR measurements over time to improve HF risk estimation in T2D.

Arterial Stiffness and Left Ventricular Diastolic Function in Subjects with Euthyroidism.

Objective: Previous literature has suggested that the cardiovascular risk factors associated with subclinical hypothyroidism (SCH) may be found in subjects with euthyroidism, but research relating to increased arterial stiffness (AS) and left ventricular (LV) diastolic dysfunction, which have been proven to exist in patients with SCH, is limited in patients with euthyroidism. The aim of this study was to investigate this. Methods: A total of 249 participants with euthyroidism were divided into two groups based on their thyroid-stimulating hormone (TSH) levels: Group A (TSH level ranging from 0.49 to 2.5 mIU/L, n = 170) and Group B (TSH level ranging from 2.5 to 4.91 mIU/L, n = 79). The Cardiovascular Profiling System through brachial-ankle pulse wave velocity (baPWV) was used to assess AS, and the LV function was evaluated using Color-Doppler-Echocardiography. The Student's unpaired t-test and Pearson's χ 2 test were conducted to compare the clinical parameters. Spearman's correlation analysis and multiple logistic regression analysis were used to analyze the association between thyroid function, baPWV, and LV diastolic function parameters. Results: Significant differences existed between the two groups in free triiodothyronine ( fT 3 ) values and systolic blood pressure (BP) (p < 0.05). When compared with Group A, the baPWV was higher, the A wave increased, and the E/A ratio was lower in Group B (p < 0.01). The multiple logistic regression analysis showed that fT 3 was associated with a higher baPWV (p < 0.001). The E/A ratio was directly correlated with TSH, fT 3 , and baPWV (p < 0.05), and diastolic BP was significantly directly correlated with the E/A ratio (p < 0.05). Thyroperoxidase antibody was not a significant variable in the regression analysis (p > 0.05). Conclusions: An association was found between thyroid function, baPWV, and the E/A ratio in subjects with euthyroidism. Further study is needed to confirm these conclusions.

The effect of sludge retention time (SRT) on the Nitrifier typical kinetics at ambient temperature under the low ammonia density.

Sludge retention time (SRT) regulation is one of the essential management techniques for refined control of the main-sidestream treatment process under the low ammonia density. It is indispensable to understand the effect of SRTs changes on the Nitrifier kinetics to obtain the functional separation of the Nitrifier and the refined control of the nitrification process. In this study, Nitrifier was cultured with conditions of 35 ± 0.5 °C, pH 7.5 ± 0.2, DO 5.0 ± 0.5 mg-O/L, and SRTs were controlled for 40 d, 20 d, 10 d, and 5 d. The net growth rate (µm), decay rate (b), specific growth rate (µ), the yield of the Nitrifier (YA), temperature parameter (TA), and inhibition coefficient (KI) have been measured and extended with the SRT decreases. Instead, the half-saturation coefficient (KS) decreased. In addition, the limited value of pH inhibition occurs (pHUL), and the pH of keeping 5% maximum reaction rate (pHLL) was in a relatively stable state. The trade of kinetics may be induced by the change of species structure of Nitrifier. The Nitrosomonas proportion was increased, and the Nitrospira was contrary with the SRT decreasing. It is a match for the functional separation of Nitrifier when SRTs was 20 d at ambient temperature under the low ammonia density. The kinetics of ammonia-oxidizing organisms (AOO) and nitrite-oxidizing organisms (NOO) in Nitrifier under different SRT conditions should be measured respectively to the refined control of the partial nitrification process in future study.

Advanced treatment of secondary effluent from wastewater treatment plant by a newly isolated microalga Desmodesmus sp. SNN1.

Secondary effluents contain considerable amounts of nitrogen and phosphorous, which if dumped untreated can cause eutrophication of the receiving water bodies. Microalgae can remove these nutrients and other pollutants from the wastewater effluents and play an effective role in the secondary effluent treatment. In this study, six microalgae strains (SNN1, SNN2, SNN3, SNN4, SNS1, and SNS2) were isolated and screened from the water and mud of Yingxue Lake of Shandong Jianzhu University, and their efficiencies for the removal of COD, NH4 +-N, TN, and TP in the secondary effluent were assessed. By comparing the growth performances and nutrient removal ability of algal strains in domestic sewage, we found that SNN1 (identified and named as Desmodesmus sp. SNN1) has the highest efficiency for biomass accumulation and sewage purification. Hence, the algal strain SNN1 was selected for further screening and optimization experiments. The strain showed higher biomass yield and better nutrient removal rate when the pH of secondary effluent was 9.0 and the initial inoculum concentration (optical density at 680 nm) of algal strain was 0.4. After 12 days of treatment, the concentrations of COD, NH4 +-N, TN, and TP in the secondary effluent were 31.79, 0.008, 8.631, and 0.069 mg/L, respectively. Therefore, SNN1 with the removal rates of 52.69% (COD), 99.99% (NH4 +-N), 89.09% (TN), and 94.64% (TP) displayed its high potential in nutrient removal. In addition, it also yielded 5.30 mg/L of chlorophyll a and 168.33 mg/L of lipids. These results demonstrated that this strain exhibited an effective treatment capacity for secondary effluent and microalgal oil production. This study is helpful to provide a strategy for the resource utilization of secondary effluent and the conservation of freshwater resources required by microalgae culture.

Association between KCNJ11 gene polymorphisms and risk of type 2 diabetes mellitus in East Asian populations: a meta-analysis in 42,573 individuals.

A number of studies have been performed to identify the association between potassium inwardly-rectifying channel, subfamily J, member 11 (KCNJ11) gene and type 2 diabetes mellitus (T2DM) in East Asian populations, with inconsistent results. The main aim of this work was to evaluate more precisely the genetic influence of KCNJ11 on T2DM in East Asian populations by means of a meta-analysis. We identified 20 articles for qualitative analysis and 16 were eligible for quantitative analysis (meta-analysis) by database searching up to May 2010. The association was assessed under different genetic models, and the pooled odds ratios (ORs) with 95% confidence intervals (95% CIs) were calculated. The allelic and genotypic contrast demonstrated that the association between KCNJ11 and T2DM was significant for rs5210. However, not all results for rs5215 and rs5218 showed significant associations. For rs5219, the combined ORs (95% CIs) for allelic contrast, dominant and recessive models contrast (with allelic frequency and genotypic distribution data) were 1.139 (1.093-1.188), 1.177 (1.099-1.259) and 1.207 (1.094-1.332), respectively (random effect model). The analysis on the most completely adjusted ORs (95% CIs) by the covariates of rs5219 all presented significant associations under different genetic models. Population-stratified analysis (Korean, Japanese and Chinese) and sensitivity analysis verified the significant results. Cumulative meta-analysis including publication time and sample size illustrated the exaggerated genetic effect in the earliest studies. Heterogeneity and publication bias were assessed. Our study verified that single nucleotide polymorphisms (SNPs) of KCNJ11 gene were significantly associated with the risk of T2DM in East Asian populations.

Effects of Antihypertensive Drugs on Thyroid Function in Type 2 Diabetes Patients With Euthyroidism.

Objective: There is little literature about whether antihypertensive drugs would affect thyroid function in patients with euthyroid type 2 diabetes, which was significant in maintaining a proper balance of thyroid function. A retrospective cohort study was conducted to evaluate the influence of antihypertensive drugs on thyroid function in patients with type 2 diabetes with euthyroidism. Design and Methods: The study involved dividing 698 patients with antihypertensive monotherapy into five groups according to the antihypertensive drugs they were treated with. Antihypertensive drugs included in this study were ß-blockers, angiotensin-converting enzyme inhibitors (ACEI), angiotensin receptor blockers (ARB), and calcium channel blockers (CCB). The clinical data and thyroid function level between or within groups were compared. Multiple logistic regression analysis was conducted to evaluate the association of antihypertensive drugs with thyroid function level. Results: Selective ß1- adrenergic receptor blockers treatment was related to thyroid-stimulating hormone (TSH), increasing in patients with diabetes and euthyroidism as shown by multiple logistic regression analysis. The association existed after adjustment for confounding factors. No significant influence on thyroid function was found among other antihypertensive drugs. Conclusion: These data show the TSH-lifting effect of selective ß1-adrenergic receptor blockers in patients with type 2 diabetes with euthyroidism.

Marinobacter daqiaonensis sp. nov., a moderate halophile isolated from a Yellow Sea salt pond.

A Gram-negative, oxidase- and catalase-positive, moderately halophilic strain, designated YCSA40(T), was isolated from sediment of Daqiao saltern in Qingdao, on the east coast of China. Growth occurred at 10-45 °C, at pH 5-9 and with 1-15% NaCl. Strain YCSA40(T) showed the highest 16S rRNA gene sequence similarity to Marinobacter segnicrescens SS011B1-4(T) (97%) and M. gudaonensis SL014B61A(T) (96.9%) and 16S rRNA gene sequence phylogenetic analysis assigned the isolate to the genus Marinobacter. Strain YCSA40(T) contained C(18:1)ω9c (34.8%), C(16:0) (11.6%), C(19:0) cyclo ω10c/C(19:1)ω6c (10.5%), C(16:1)ω9c (8.4%), C(17:0) (6.3%) and C(12:0) 3-OH (5.8%) as the predominant fatty acids. The DNA G+C content was 60.8 mol% and the major ubiquinone was Q-9. These chemotaxonomic characters were all consistent with membership of the genus Marinobacter. DNA-DNA relatedness between the isolate and M. segnicrescens CGMCC 1.6489(T), M. gudaonensis CGMCC 1.6294(T) and other type strains of species of the genus Marinobacter was ≤30%. On the basis of the aforementioned data, it was concluded that strain YCSA40(T) represents a novel species of the genus Marinobacter, for which the name Marinobacter daqiaonensis sp. nov. is proposed. The type strain is YCSA40(T) (=CGMCC 1.9167(T) =NCCB 100308(T) =LMG 25365(T)).

Sunxiuqinia elliptica gen. nov., sp. nov., a member of the phylum Bacteroidetes isolated from sediment in a sea cucumber farm.

Three novel aerobic, elliptic bacteria, designated DQHS4(T), DQHS8 and DQHS15, were isolated from sediment of a seashore pond for sea cucumber culture in Jimo, Qingdao, on the east coast of China. Cells were Gram-, oxidase- and catalase-negative. All three strains grew at 15-42 °C, pH 5-9 and NaCl concentrations between 0.5 and 10%. DNA-DNA hybridization experiments revealed high (>85%) relatedness among the three novel isolates and suggested that the strains constitute a single species. Comparative 16S rRNA gene sequence analysis indicated that these bacteria had less than 90% similarity to all described species of the phylum Bacteroidetes; the closest relative of the three isolates was Prolixibacter bellariivorans F2(T), sharing only 89.6% sequence similarity. The major cellular fatty acids were iso-C(17:0) 3-OH (19.8-20.0%), iso-C(15:0) (16.9-17.3%), anteiso-C(17:1) B and/or iso-C(17:1) I (7.4-8.7%), C(17:0) 2-OH (8.4%), anteiso-C(15:0) (8.2-8.6%) and C(17:1)ω6c (5.6-6.0%). The major respiratory quinone was menaquinone-7 (MK-7) and the DNA G+C content was 41.8-43.5 mol%. Based on the distinct phylogenetic position and the combination of genotypic, phenotypic and chemotaxonomic characteristics, these three strains were considered to represent a novel species of a new genus in the phylum Bacteroidetes, for which the name Sunxiuqinia elliptica gen. nov., sp. nov. is proposed. The type strain of Sunxiuqinia elliptica is DQHS4(T) (=CGMCC 1.9156(T) =NCCB 100301(T) =LMG 25367(T)).

Halomonas daqiaonensis sp. nov., a moderately halophilic, denitrifying bacterium isolated from a littoral saltern.

Two novel gram-negative, oxidase- and catalase-positive, rod-shaped bacterial strains, designated YCSA28(T) and YCSA39, were isolated from sediment of Daqiao saltern, Jimo, Qingdao, on the east coast of China. The two strains grew optimally at 28-30 °C, at pH 7.5 and in the presence of 7-8 % (w/v) NaCl. They were assigned to the genus Halomonas, class Gammaproteobacteria, based on 16S rRNA gene sequence analysis. The major cellular fatty acids of the two strains were C(18 : 1)ω7c (42.9 %), C(16 : 0) (23.1 %) and C(16 : 1)ω7c/ω6c (18.0 %), and Q-9 was the major ubiquinone. The G+C content of the DNA of strains YCSA28(T) and YCSA39 was 63.7 and 63.9 mol%, respectively. The predominant respiratory lipoquinone, cellular fatty acid profiles and DNA G+C content of strains YCSA28(T) and YCSA39 were consistent with those of recognized species of the genus Halomonas. Levels of DNA-DNA relatedness between strains YCSA28(T) and YCSA39, between YCSA28(T) and Halomonas ventosae Al12(T), and between YCSA39 and H. ventosae Al12(T) were 95, 45 and 50 %, respectively. Together, these data indicated that strains YCSA28(T) and YCSA39 represent a single novel species of the genus Halomonas, for which the name Halomonas daqiaonensis sp. nov. is proposed. The type strain is YCSA28(T) ( = CGMCC 1.9150(T)  = NCCB 100305(T)  = MCCC 1B00920(T)).

Cohaesibacter marisflavi sp. nov., isolated from sediment of a seawater pond used for sea cucumber culture, and emended description of the genus Cohaesibacter.

A Gram-negative, catalase-negative, oxidase-positive, rod-shaped bacterium, strain DQHS21(T), was isolated from sediment of a seawater pond used for sea cucumber culture at Jimo in Qingdao province on the east coast of China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain DQHS21(T) belonged to the genus Cohaesibacter, sharing the highest sequence similarity (96.1 %) with Cohaesibacter gelatinilyticus CL-GR15(T), while the similarity to other strains was below 93.0 %. The cellular fatty acids consisted mainly of C(18 : 1)ω7c (60.7 %), C(18 : 0) (17.8 %), C(16 : 0) (8.5 %) and summed feature 3 (C(16 : 1)ω7c and/or iso-C(15 : 0) 2-OH; 6.0 %), which together accounted for 93 % of the total fatty acids. Ubiquinone 10 was the major quinone. The G+C content of the chromosomal DNA of strain DQHS21(T) was 55.2 mol%. The combined genotypic and phenotypic data showed that strain DQHS21(T) represents a novel species of the genus Cohaesibacter, for which the name Cohaesibacter marisflavi sp. nov. is proposed, with the type strain DQHS21(T) ( = CGMCC 1.9157(T)  = NCCB 100300(T)).

Analysis of the genetic diversity of the lymphocystis virus and its evolutionary relationship with its hosts.

Lymphocystis disease virus (LCDV) is the causative agent of lymphocystis disease. In this study, the mcp gene of LCDV and the cyt b gene of the host fish were selected as molecular markers, and the phylogenetic relationships between LCDV and its host were analyzed. The 25 LCDV isolates examined in this study were attributed to seven LCDV genotypes: genotype I (LCDV-1), genotype II (LCDV-cn, etc.), genotype III (LCDV-rf), genotype IV (LCDV-rc and LCDV-sb), genotype V (LCDV-cb), genotype VI (LCDV-tl), and genotype VII (LCDV-sa). Genotype VII is a new genotype. LCDV1 was found to have differentiated first, followed by LCDV-rf; then LCDV-tl; LCDV-cb; and then LCDV-sa; and by LCDV-rc and LCDV-sb; and finally by LCDV-cn, LCDV-C, and LCDV-jf. From the host evolutionary perspective, Rachycentron canadum was found to have differentiated first, followed by Trichogaster leeri, Chanda baculis, and Sebastes schlegeli, Lateolabrax sp., Sparus aurata, Platichthys flesus, and Paralichthys olivaceus. Comparison of the phylogenies of the host fish species and LCDVs revealed no significant evidence of cospeciation between LCDVs and their host fish. In-depth studies of the genetic variation in LCDVs can enhance our understanding of the mechanism of LCDV infection, which may provide important insights into the prevention and treatment of lymphocystis disease.

Isolation and Characterization of a Phosphate-Solubilizing Halophilic Bacterium Kushneria sp. YCWA18 from Daqiao Saltern on the Coast of Yellow Sea of China.

Phosphate-solubilizing bacteria (PSB) function in soil phosphorus cycle, increasing the bioavailability of soil phosphorus for plants. Isolation and application of salt-tolerant or halophilic PSB will facilitate the development of saline-alkali soil-based agriculture. A moderately halophilic bacterium was isolated from the sediment of Daqiao saltern on the eastern coast of China, which also performs phosphate-solubilizing ability. The bacterium was assigned to genus Kushneria according to its 16S rRNA gene sequence, and accordingly named as Kushneria sp. YCWA18. The fastest growth was observed when the culturing temperature was 28°C and the concentration of NaCl was 6% (w/v). It was founds that the bacterium can survive at a concentration of NaCl up to 20%. At the optimum condition, the bacterium solubilized 283.16 µg/mL phosphorus in 11 days after being inoculated in 200 mL Ca(3)(PO(4))(2) containing liquid medium, and 47.52 µg/mL phosphorus in 8 days after being inoculated in 200 mL lecithin-containing liquid medium. The growth of the bacterium was concomitant with a significant decrease of acidity of the medium.

Immune Efficacy of a Genetically Engineered Vaccine against Lymphocystis Disease Virus: Analysis of Different Immunization Strategies.

Here, we report the construction of a vaccine against lymphocystis disease virus (LCDV) using nucleic acid vaccination technology. A fragment of the major capsid protein encoding gene from an LCDV isolated from China (LCDV-cn) was cloned into an eukaryotic expression vector pEGFP-N2, yielding a recombinant plasmid pEGFP-N2-LCDV-cn0.6 kb. This plasmid was immediately expressed after liposomal transfer into the Japanese flounder embryo cell line. The recombinant plasmid was inoculated into Japanese flounder via two routes (intramuscular injection and hypodermic injection) at three doses (0.1, 5, and 15 µg), and then T-lymphopoiesis in different tissues and antibodies raised against LCDV were evaluated. The results indicated that this recombinant plasmid induced unique humoral or cell-mediated immune responses depending on the inoculation route and conferred immune protection. Furthermore, the humoral immune responses and protective effects were significantly increased at higher vaccine doses via the two injection routes. Plasmid pEGFP-N2-LCDV0.6 kb is therefore a promising vaccine candidate against LCDV in Japanese flounder.

Intensive Risk Factor Management and Cardiovascular Autonomic Neuropathy in Type 2 Diabetes: The ACCORD Trial.

OBJECTIVE: The effects of preventive interventions on cardiovascular autonomic neuropathy (CAN) remain unclear. We examined the effect of intensively treating traditional risk factors for CAN, including hyperglycemia, hypertension, and dyslipidemia, in individuals with type 2 diabetes (T2D) and high cardiovascular risk participating in the Action to Control Cardiovascular Risk in Diabetes (ACCORD) trial. RESEARCH DESIGN AND METHODS: CAN was defined as heart rate variability indices below the fifth percentile of the normal distribution. Of 10,251 ACCORD participants, 71% (n = 7,275) had a CAN evaluation at study entry and at least once after randomization. The effects of intensive interventions on CAN were analyzed among these subjects through generalized linear mixed models. RESULTS: As compared with standard intervention, intensive glucose treatment reduced CAN risk by 16% (odds ratio [OR] 0.84, 95% CI 0.75-0.94, P = 0.003)-an effect driven by individuals without cardiovascular disease (CVD) at baseline (OR 0.73, 95% CI 0.63-0.85, P < 0.0001) rather than those with CVD (OR 1.10, 95% CI 0.91-1.34, P = 0.34) (P interaction = 0.001). Intensive blood pressure (BP) intervention decreased CAN risk by 25% (OR 0.75, 95% CI 0.63-0.89, P = 0.001), especially in patients ≥65 years old (OR 0.66, 95% CI 0.49-0.88, P = 0.005) (P interaction = 0.05). Fenofibrate did not have a significant effect on CAN (OR 0.91, 95% CI 0.78-1.07, P = 0.26). CONCLUSIONS: These data confirm a beneficial effect of intensive glycemic therapy and demonstrate, for the first time, a similar benefit of intensive BP control on CAN in T2D. A negative CVD history identifies T2D patients who especially benefit from intensive glycemic control for CAN prevention.

Implication of genetic variants near SLC30A8, HHEX, CDKAL1, CDKN2A/B, IGF2BP2, FTO, TCF2, KCNQ1, and WFS1 in type 2 diabetes in a Chinese population.

BACKGROUND: Recently, several genome-wide and candidate gene association studies have identified many novel genetic loci for type 2 diabetes (T2D); among these genes, CDKAL1, IGF2BP2, SLC30A8, CDKN2A/B, HHEX, FTO, TCF2, KCNQ1, and WFS1 are the most important. We aimed to determine the effects of these genetic loci associated with T2D in the Chinese Han population of China. METHODS: Single-nucleotide polymorphisms (SNPs) in or near CDKAL1, IGF2BP2, SLC30A8, CDKN2A/B, HHEX, FTO, TCF2, KCNQ1, and WFS1 genes were genotyped in a case-control Chinese Han sample living in Beijing, China involving 1024 patients with T2D and 1005 control subjects. RESULTS: In Chinese Han, we replicated the associations between 7 genetic loci and T2D, with risk allele-specific odds ratios (ORs) as follows: 1.27 (95% CI, 1.11-1.45; p = 0.0008) for CDKAL1-rs10946398, 1.26 (95% CI, 1.08-1.47; p = 0.003) for IGF2BP2-rs4402960, 1.19 (95% CI, 1.04-1.37; p = 0.009) for SLC30A8-rs13266634, 1.22 (95% CI, 1.06-1.41; p = 0.005) for CDKN2A/B-rs10811661, 1.20 (95% CI, 1.01-1.42; p = 0.03) for HHEX-rs5015480, 1.37 (95% CI, 1.19-1.69; p = 1.0 x 10(-4)) for KCNQ1-rs2237892, and 1.24 (95% CI, 1.01-1.52; p = 0.046) for FTO-rs8050136 after adjustment for age, gender, and body mass index. Not only did an association between WFS1-rs6446482 and early-onset T2D exist in the subgroup analysis, but TCF2-rs7501939 and WFS1-rs6446482 were also confirmed to confer risk for T2D in this meta-analysis. Moreover, the relationship between FTO-rs8050136 and body mass index, together with the effect of CDKAL1-rs10946398 on beta cell function, was also observed in the control individuals. CONCLUSIONS: Our findings support the important contribution of these genetic loci to susceptibility for T2D in the Chinese Han population in Beijing of China.

Identification of the pathogens associated with skin ulceration and peristome tumescence in cultured sea cucumbers Apostichopus japonicus (Selenka).

The aquaculture of sea cucumbers Apostichopus japonicus (Selenka) has developed rapidly in China in recent years, but is increasingly affected by diseases such as skin ulceration and peristome tumescence. Previous studies on the pathogens causing these diseases focused largely on bacterial causes. In December 2008, we isolated four dominant bacterial species from lesions present in A. japonicus with the aforementioned diseases, from a farm in Yangkou (Qingdao, China). With two of these bacterial species, experimental infection of healthy A. japonicus resulted in the same disease symptoms that occurred in naturally infected A. japonicus. These two species were identified as Pseudoalteromonas sp. and Pseudoalteromonas tetraodonis. The early symptoms of infection for these bacterial species were ulcer spots on the dorsal skin and abdominal parapodia, followed by an increase in the number of ulcer spots or their merging into larger spots. Additionally, we isolated a spherical virus 100-250nm in diameter and with a bilayer capsule, from A. japonicus with another disease from four different farms. By experimental infection with crude extracts of the virus, healthy laboratory-acclimatized A. japonicus developed the same symptoms as in natural infected cases. The early symptoms of viral infection comprised a decrease in tentacle activity, decay of dorsal papillate podia, peristome tumescence and abdominal ulceration. Our study demonstrates that the bacteria and virus were both responsible for skin ulceration and peristome tumescence in A. japonicus, but resulted in different early disease symptoms.

The formulation and immunisation of oral poly(DL-lactide-co-glycolide) microcapsules containing a plasmid vaccine against lymphocystis disease virus in Japanese flounder (Paralichthys olivaceus).

Nucleic acid-based immunotherapy is a new treatment option for fish immunisation in intensive culture. However, DNA-based vaccines would be hydrolyzed or denaturized because of the existence of nucleases and severe gastrointestinal conditions. Poly(DL-lactide-co-glycolide) (PLGA) microcapsules, loaded with plasmid DNA (pDNA) against lymphocystis disease virus (LCDV), were prepared by modified water in oil in water (W/O/W) double emulsion method in our laboratory. Encapsulation efficiency, loading percent and diameter of microcapsules were 78-88%, 0.5-0.7% and less than 10 mum, respectively. In simulated gastric fluid (SGF), less than 10% of pDNA was released from microcapsules in 12 h, and about 6.5% of pDNA was released in 12 h in simulated intestinal fluid (SIF). The content of the supercoiled of pDNA in microcapsules and control was 80% and 89% respectively, which indicated that a little supercoiled pDNA degradation occurred during encapsulation. RT-PCR showed that lots of RNA containing information of MCP gene existed in all tissues of fish vaccinated with microcapsules 10-90 days after oral administration. SDS-PAGE and immunoblots, as well as immunofluorescence images, displayed that major capsid protein (MCP) of LCDV was expressed in tissues of fish vaccinated with pDNA-loaded microcapsules. In addition, indirect enzyme-linked immunosorbent assay (ELISA) showed that the immune responses of sera were positive (O.D> or =0.3) from week 1 to week 24 for fish vaccinated with microcapsules, in comparison with fish vaccinated with naked pDNA. Our results suggested that PLGA microcapsules were promising oral carriers for pDNA delivery. This encapsulation technique had potential for drug delivery applications due to its ease of operation and notable immunisation efficacy.

Chitosan microspheres as candidate plasmid vaccine carrier for oral immunisation of Japanese flounder (Paralichthys olivaceus).

Oral DNA-based immunotherapy is a new treatment option for fish immunisation in intensive culture. However, because of the existence of the nucleases and severe gastrointestinal conditions, DNA-based vaccines can be hydrolyzed or denatured. In our laboratory, a plasmid DNA (pDNA) containing major capsid protein (MCP) gene of lymphocystis disease virus (LCDV) was prepared, and then pDNA was encapsulated in chitosan microspheres through an emulsion-based methodology. The yield, loading percent and encapsulation efficiency of microspheres were 93.6%, 0.3% and 94.5%, respectively. Scanning electron microscopy (SEM) showed that pDNA-loaded microspheres yielded a spherical shape with smooth surfaces. The disproportion of super-coiled to open circle and linear pDNA suggested that high transfection efficiencies of pDNA in microspheres were retained. The cumulative release of pDNA showed that chitosan microspheres were resistant to degradation in simulated gastrointestinal tract environment. The release profile at PBS buffer (pH 7.4) displayed that pDNA-loaded chitosan microspheres had a release up to 42 days after intestinal imbibition. RT-PCR showed that RNA containing information of MCP gene existed in various tissues 10-90 days post-vaccination. SDS-PAGE and immunofluorescent images indicated that pDNA expressed MCP in tissues of fish 10-90 days after oral administration. In addition, indirect ELISA displayed that the immune responses of sera were positive (O.D.> or =0.3) from week 1 to week 16 for fish vaccinated with microspheres, in comparison with fish vaccinated with naked pDNA. Data obtained suggested that chitosan microspheres were promising carriers for oral pDNA vaccine. Because this encapsulation technique was easy to operate and immunisation efficacy of microspheres loaded with pDNA was significant, it had potential to be used in drug delivery applications.

Meta-analysis on autologous stem cell transplantation in the treatment of limb ischemic.

AIM: Meta-analysis on the effectiveness of the autologous stem cell transplantation in curing limb ischemic. METHOD: Consulting papers relate to the autologous stem cell transplantation in curing limb ischemic in PubMed, CNKI, Wan-fang Data and VIP. Based on include and exclude standards, we arrange at least 2 evaluators sifting these papers separately, doing Quality evaluation and information extraction and then cross checking. Negotiate through a third party if any disagreement comes out. Review Manager 4.2 is used in Meta-analysis. RESULT: Involved 7 papers, all in English version. Patients involved all diagnosed as critical limb ischemia (CLI). Results show that no adverse reaction occurred during this study. Amputation rate in patients with stem cell group treatment is lower than control group (P < 0.05). And no notable difference in improving ABI (P > 0.05). CONCLUSIONS: Applying autologous stem cell transplantation in curing limb ischemic does not have obviously effectiveness in the improvement of ABI of the limb ischemic patients. But it can dramatically reduce the rate of amputation. So autologous stem cell transplantation is a good and safe choice for patients have no choice but amputation.

Rs4074134 near BDNF gene is associated with type 2 diabetes mellitus in Chinese Han population independently of body mass index.

Obesity and family history are the most important predictors for type 2 diabetes mellitus(T2DM) in the Chinese Han population. However, it is not known whether the genetic loci related to obesity are associated with the risk of developing T2DM in this population. The present case-control study evaluated the associations between five genetic loci for obesity and the pathogenesis of T2DM. The study included 1117 Chinese Han patients with T2DM, 1629 patients with pre-diabetes (impaired fasting glucose and impaired glucose tolerance, IFG/IGT) and 1113 control subjects residing in Beijing. Five genetic loci including rs2815752 near NEGR1, rs10938397 near GNPDA2, rs4074134 near BDNF, rs17782313 near MC4R and rs1084753 near KCTD15 were genotyped. The results showed an association between rs4074134-BDNF minor allele and T2DM irrespective of age, gender and body mass index (BMI) (OR = 0.87; 95%CI: 0.77-0.99, P = 0.04). This SNP was also associated with pre-diabetes (OR = 0.87; 95%CI: 0.77-0.97, P = 0.01) independently of age, gender and BMI. No associations were found between diabetes or pre-diabetes and any of the other SNP loci studied. Genotype-phenotype association analysis (adjusting for age and gender) showed rs4074134-BDNF to be associated with BMI, waist circumference, fasting and postprandial plasma glucose, fasting serum insulin, and HOMA-IR in subjects without T2DM. However, fasting and postprandial plasma glucose were the only significant factors after adjusting for BMI. These results suggest that the common variation of BDNF (rs4074134) is associated with T2DM independently of obesity in Chinese Han population. This variant also has an effect on plasma glucose concentration, BMI and insulin sensitivity.