Assessment of sulfonamide contamination in aquatic environments: A first report for Argentina and environmental risk assessment.

The global surge in pharmaceutical consumption, driven by increasing population and the demand for animal proteins, leads to the discharge of diverse pollutants, including antibiotic residues, into water bodies. Sulfonamides, being water-soluble compounds, can readily enter surface run-off, posing potential risks to non-target species despite their low environmental concentrations. Latin America has implemented intensive production systems highly dependent on antimicrobials for productivity and animal health, yet there is a paucity of information regarding their concentration in the region. The objective of this study was to evaluate the presence of sulfonamides in water and sediment samples and assess their potential ecological risks through an environmental risk assessment. The Río de la Plata basin collects the waters of the Paraguay, Paraná, and Uruguay rivers, together with their tributaries and various wetlands, passing through the provinces in Argentina known for their significant animal husbandry production. Two sampling campaigns were carried out for sediment, while only one campaign was conducted for surface waters. The samples were analyzed by high performance liquid chromatography tandem mass-spectrometry (HPLC-MS/MS). None of the examined sulfonamide antibiotics were detected in the sediment samples from both sampling campaigns. In contrast, sulfadiazine (95 %), sulfamethoxazole (91 %), and sulfathiazole (73 %) were detected in the water samples. Sulfadiazine was found in the concentration range of 8 to 128 ng/L, while sulfamethoxazole and sulfathiazole were observed at concentrations ranging from 3.0 to 32.5 ng/L and 2.9 to 8.1 ng/L, respectively. Based on the environmental risk assessment conducted using the sulfonamide concentrations, most samples indicated a medium risk for aquatic biota, with only one sample surpassing the high-risk threshold. This study represents the first report presenting data on the presence of sulfonamide antibiotics in the aquatic environment of Argentina.

Provider delay in treatment initiation and its influence on survival outcomes in women with operable breast cancer.

AIM: The goal of this study was to determine whether a delay in starting treatment via surgery or neoadjuvant chemotherapy is related to a decrease in cancer-specific survival (CSS) in women with operable breast cancer (BrCr). BACKGROUND: Limited medical infrastructure and a lack of cancer prevention awareness in low- and middle-income countries have caused high BrCr incidence and mortality rates. METHODS: We analyzed a retrospective cohort of 720 women treated at a single center from 2005 to 2012. CSS estimates were obtained by the Kaplan-Meier method. A Cox model of proportional risks was performed to obtain the risk of dying from BrCr. We also obtained the risk according to the category of treatment initiation. RESULTS: Women with locally advanced stages and without hormone receptor expression were more likely to initiate treatment after 45 days. Patients in Stage IIIA had a 78.1% survival if treatment was initiated before 45 days (95% CI, 0.70-0.84) and 63.6% survival if treatment was started after 45 days (95% CI, 0.44-0.78; p < 0.001). Patients in Stage IIIB had a 62.9% survival if treatment was initiated before 45 days (95% CI, 0.53-0.72) and 57.4% survival if treatment started after 45 days (95% CI, 0.31-0.89; p < 0.001). Prognostic factors in which lower survival was recognized were Stage IIIA, Stage IIIB, treatment initiation after 45 days, and triple-negative tumors. CONCLUSIONS: The initiation of treatment within the first 45 days of diagnosis of BrCr in women portends better survival compared with those who began treatment longer than 45 days from diagnosis.

Determinación del estado nutricional mediante el ángulo de fase en pacientes en hemodiálisis.

INTRODUCTION: Multi-frequency bio-impedance analysis (BIA) accurately evaluates body composition. OBJECTIVE: To assess nutritional status and its relationship with body mass index (BMI) or with BIA-obtained phase angle in patients with end-stage renal disease (ESRD) on hemodialysis. METHOD: BMI, BIA and dialysis malnutrition score (DMS) were assessed in 99 patients on hemodialysis (43.6 ± 17.2 years of age, 58.8% males). Multivariate linear regression analysis was used to determine BMI and phase angle independent associations with DMS. RESULTS: Malnutrition risk (DMS > 13) showed a gradual increase among patients with normal BMI and phase angle (44.4%), low BMI (45.8%), low phase angle (64.0%), and in those with both parameters at abnormal ranges (68.0%). The phase angle was the only variable that was independently associated with patient nutritional status (standardized coefficient beta -0.372, p < 0.001), accounting for 13.8% of DMS variation. CONCLUSION: Phase angle is inversely and independently associated with malnutrition risk in patients with ESRD, which suggests that BIA-assessed body composition might be better than BMI in the clinical assessment of patients with ESRD.

INTRODUCCIÓN: La bioimpedancia eléctrica multifrecuencia (BIE) evalúa con precisión la composición corporal. OBJETIVO: Investigar el estado nutricional y su relación con el índice de masa corporal (IMC) o el ángulo fase obtenido por BIE en pacientes con enfermedad renal terminal (ERT) en hemodiálisis. MÉTODO: En 99 pacientes en hemodiálisis (43.6 ± 17.2 años, 58.8 % varones) se evaluó IMC, BIE y puntuación de desnutrición (DMS, Dialysis Malnutrition Score). La independencia de las asociaciones del IMC y el ángulo fase con el DMS se determinó con regresión lineal múltiple. RESULTADOS: El riesgo de desnutrición (DMS > 13) se incrementó gradualmente entre los sujetos con IMC y ángulo fase normales (44.4 %), IMC bajo (45.8 %), ángulo fase bajo (64.0 %) y en aquellos con ambos parámetros anormales (68.0 %). El ángulo fase fue la única variable asociada independientemente con el estado nutricional de los pacientes (coeficiente beta estandarizado −0.372, p < 0.001), lo que explicó 13.8 % de la variación en el DMS. CONCLUSIONES: El ángulo fase se asocia inversa e independientemente con el riesgo de desnutrición en pacientes con ERT, lo que sugiere que la composición corporal evaluada por BIE pudiera ser mejor que el IMC en la valoración clínica de pacientes con esta enfermedad.

Determinación del estado nutricional mediante el ángulo de fase en pacientes en hemodiálisis / Determination of nutritional status by phase angle in hemodialysis patients

Resumen Introducción: La bioimpedancia eléctrica multifrecuencia (BIE) evalúa con precisión la composición corporal. Objetivo: Investigar el estado nutricional y su relación con el índice de masa corporal (IMC) o el ángulo fase obtenido por BIE en pacientes con enfermedad renal terminal (ERT) en hemodiálisis. Método: En 99 pacientes en hemodiálisis (43.6 ± 17.2 años, 58.8 % varones) se evaluó IMC, BIE y puntuación de desnutrición (DMS, Dialysis Malnutrition Score). La independencia de las asociaciones del IMC y el ángulo fase con el DMS se determinó con regresión lineal múltiple. Resultados: El riesgo de desnutrición (DMS > 13) se incrementó gradualmente entre los sujetos con IMC y ángulo fase normales (44.4 %), IMC bajo (45.8 %), ángulo fase bajo (64.0 %) y en aquellos con ambos parámetros anormales (68.0 %). El ángulo fase fue la única variable asociada independientemente con el estado nutricional de los pacientes (coeficiente beta estandarizado −0.372, p < 0.001), lo que explicó 13.8 % de la variación en el DMS. Conclusiones: El ángulo fase se asocia inversa e independientemente con el riesgo de desnutrición en pacientes con ERT, lo que sugiere que la composición corporal evaluada por BIE pudiera ser mejor que el IMC en la valoración clínica de pacientes con esta enfermedad.

Abstract Introduction: Multi-frequency bio-impedance analysis (BIA) accurately evaluates body composition. Objective: To assess nutritional status and its relationship with body mass index (BMI) or with BIA-obtained phase angle in patients with end-stage renal disease (ESRD) on hemodialysis. Method: BMI, BIA and dialysis malnutrition score (DMS) were assessed in 99 patients on hemodialysis (43.6 ± 17.2 years of age, 58.8% males). Multivariate linear regression analysis was used to determine BMI and phase angle independent associations with DMS. Results: Malnutrition risk (DMS > 13) showed a gradual increase among patients with normal BMI and phase angle (44.4%), low BMI (45.8%), low phase angle (64.0%), and in those with both parameters at abnormal ranges (68.0%). The phase angle was the only variable that was independently associated with patient nutritional status (standardized coefficient beta −0.372, p < 0.001), accounting for 13.8% of DMS variation. Conclusion: Phase angle is inversely and independently associated with malnutrition risk in patients with ESRD, which suggests that BIA-assessed body composition might be better than BMI in the clinical assessment of patients with ESRD.

Liposomes can both enhance or reduce drugs penetration through the skin.

The adequate formulation of topical vehicles to treat skin diseases is particularly complex. A desirable formulation should enhance the accumulation of the active drugs in the target tissue (the skin), while avoiding the penetration enhancement to be so large that the drugs reach the systemic circulation in toxic amounts. We have evaluated the transcutaneous penetration of three drugs chosen for their widely variable physicochemical properties: Amphotericin B, Imiquimod and Indole. We incorporated the drugs in fluid or ultra-flexible liposomes. Ultra-flexible liposomes produced enhancement of drug penetration into/through human skin in all cases in comparison with fluid liposomes without detergent, regardless of drug molecular weight. At the same time, our results indicate that liposomes can impede the transcutaneous penetration of molecules, in particular small ones.

Genetic and nongenetic profiling of milk pregnancy-associated glycoproteins in Holstein cattle.

Pregnancy-associated glycoproteins (PAG) are secreted by the trophoblast and are detectable in maternal circulation around the time of attachment of the fetal placenta, as well as in blood and milk throughout gestation. The understanding of the genetic mechanisms controlling PAG levels can confer advantages for livestock breeding programs given the precocity and the ease of obtaining this phenotype from routine pregnancy diagnosis. The aim of this study was to characterize PAG levels by estimating genetic parameters and correlations with other dairy traits, and to identify genomic regions and candidate genes associated with PAG levels in milk. The PAG data consisted of pregnancy diagnoses using commercial assays from 2012 to 2017, and genotype data consisted of 54,123 SNP markers for 2,352 individuals (embryos and dams). The model included contemporary group (herd, year, and season) and embryo age as fixed effects, and random embryonic (direct) and maternal (indirect) genetic effects. Using genomic data, the estimated heritability for direct and maternal genetic effects (± standard deviations) were 0.23 ± 0.05 and 0.11 ± 0.05, respectively. The genetic correlation between these effects was almost zero (0.001 ± 0.06). A preliminary analysis revealed low correlations between milk PAG levels and other dairy traits. The genome-wide association analysis was performed using 2 approaches: single-marker and single-step using all markers. Four genomic regions with direct genetic effects were detected on Bos taurus autosome (BTA) 6, BTA7, BTA19, and BTA29 of the embryonic genome. The BTA29 locus was within the bovine PAG gene cluster, suggesting a cis-regulatory quantitative trait locus on the PAG expression. However, other associations, without an obvious link to PAG expression, could be related to the transportation of PAG and their concentration in milk. Only 1 region from the maternal genome, on BTA4, had a significant indirect effect, where WNT2 is a candidate gene related to placenta vascularization and gestation health. Collectively, our results suggest a moderate genetic control of milk PAG levels from both maternal and fetal genomes, but larger studies are needed to fully evaluate the usefulness of milk PAG in the genetic evaluation of fetal growth and cow fertility.

Isolation and characterization of polymorphic microsatellite markers for the chub mackerel (Scomber japonicus) and cross-species amplification in the blue mackerel (S. australasicus).

In this study, 10 polymorphic microsatellite markers were developed in Scomber japonicus and were examined on 30 individuals collected from the North Pacific. The number of alleles per locus ranged from 4 to 17. The observed and expected heterozygosities per locus ranged from 0.2759 to 0.8621 and from 0.43071 to 0.9177, respectively. The polymorphism information content (PIC) was from 0.3931 to 0.8939. One locus showed moderate polymorphism (0.25 < PIC < 0.5), while the rest were highly polymorphic (PIC > 0.5). Two loci showed significant deviation from Hardy-Weinberg equilibrium after Bonferroni corrections (P < 0.005). No linkage disequilibrium was detected among the loci. Results of cross-species amplification showed that 10 microsatellite markers were successfully amplified in 29 individuals of S. australasicus and 9 indicated polymorphisms. These markers will be useful for investigating the genetic structure, gene flow, and species identification of S. japonicus and S. australasicus, its closely related species.

Effect of 1,25(OH2D3 on the proliferation of human mesangial cells and their expression of Ki67.

Previous studies have found that 1,25-dihydroxyvitamin D3 [1,25(OH)2D3 or VD3] exerts many biological effects, including the inhibition of cell proliferation and induction of apoptosis, but its mechanism of action remains unclear. The goal of our investigation was to explore the effects of 1,25(OH)2D3 on the proliferation of cultured human mesangial cells and their expression of Ki67 in vitro, and to establish its mechanism of action. Cultured human mesangial cells were randomly divided into the following four groups: normal control (N group; administered Dulbecco's modified Eagle's medium containing 5% fetal bovine serum), proliferation [epidermal growth factor (EGF) group; administered 10 µg/L EGF], VD3 intervention [administered 10-8 M 1,25(OH)2D3], and proliferation and intervention [EGF+VD3 group; administered 10 µg/L EGF and 10-8 M 1,25(OH)2D3]. Cells were incubated for 48 h with the corresponding treatment, and fluorescence immunocytochemistry and reverse transcription-quantitative polymerase chain reaction were used to detect expression of Ki67 protein and mRNA, respectively. Compared to the N group, Ki67 levels were found to be higher in the EGF group but significantly lower in the VD3 intervention group. Moreover, expression of Ki67 by cells in the EGF+VD3 group was significantly lower than that of those in the EGF group. All of these differences were statistically significant (P < 0.05). In conclusion, 1,25(OH)2D3 inhibited Ki67 expression and the proliferation of human mesangial cells; therefore, Ki67 may be regarded as a potent therapeutic target in mesangial proliferative glomerulonephritis.

Identification of Magnolia wufengensis (Magnoliaceae) cultivars using phenotypic traits, SSR and SRAP markers: insights into breeding and conservation.

A combination of phenotypic characterization and molecular markers may provide reliable information on new plant varieties and elucidate the conservation status of rare species. Five newly developed Magnolia wufengensis cultivars, an endangered plant species endemic to Hubei Province, China, possess more distinctive phenotypes than common Magnolia cultivars. With reference to a wild species population of M. wufengensis and a population of Magnolia denudata, morphological traits of flower organs, simple sequence repeat (SSR), and sequence-related amplified polymorphism (SRAP) markers were used. In the morphological study, six traits of floral organs were investigated and their relationships were analyzed between cultivars. In the genetic study, 9 SSR primer pairs and 10 SRAP primer combinations were screened. The five cultivars maintained a high level of genetic diversity. Genetic diversity of each M. wufengensis cultivar was much lower than that of the wild population, but was slightly higher than that of the M. denudata population. Analysis of molecular variance (AMOVA) revealed that genetic variation among populations was 20% (SRAP) and 30% (SSR), which showed a high degree of genetic differentiation among populations of the five cultivars. The dendrograms illustrated a clear separation between M. wufengensis populations and outer species, and identified two major groups among cultivars. Correlation analysis indicated a good fit between the two marker systems, but a relatively low fit between morphological and genetic traits (SRAP: r = 0.60, SSR: r = 0.52). These findings provide reliable references for the application of these molecular markers in the breeding and conservation of M. wufengensis.

Regulation of semaphorin 4D expression and cell proliferation of ovarian cancer by ERalpha and ERbeta.

Ovarian cancer is one of the most common malignancies in women. Semaphorin 4D (sema 4D) is involved in the progress of multiple cancers. In the presence of estrogen-like ligands, estrogen receptors (ERα and ERß) participate in the progress of breast and ovarian cancers by transcriptional regulation. The aim of the study was to investigate the role of sema 4D and elucidate the regulatory pattern of ERα and ERß on sema 4D expression in ovarian cancers. Sema 4D levels were up-regulated in ovarian cancer SKOV-3 cells. Patients with malignant ovarian cancers had significantly higher sema 4D levels than controls, suggesting an oncogene role of sema 4D in ovarian cancer. ERα expressions were up-regulated in SKOV-3 cells compared with normal ovarian IOSE80 epithelial cells. Conversely, down-regulation of ERß was observed in SKOV-3 cells. Forced over-expression of ERα and ERß in SKOV-3 cells was manipulated to establish ERα+ and ERß+ SKOV-3 cell lines. Incubation of ERα+ SKOV-3 cells with ERs agonist 17ß-estradiol (E2) significantly enhanced sema 4D expression and rate of cell proliferation. Incubated with E2, ERß+ SKOV-3 cells showed lower sema 4D expression and cell proliferation. Blocking ERα and ERß activities with ICI182-780 inhibitor, sema 4D expressions and cell proliferation of ERα+ and ERß+ SKOV-3 cells were recovered to control levels. Taken together, the data showed that sema 4D expression was positively correlated with the progress of ovarian cancer. ERα positively regulated sema 4D expression and accelerated cell proliferation. ERß negatively regulated sema 4D expression and inhibited cell multiplication.

Distribution of tumor stage and initial treatment modality in patients with primary hepatocellular carcinoma.

OBJECTIVE: This study reviewed the distribution of each tumor stage and each type of initial treatment modality among patients with primary hepatocellular carcinoma (HCC) treated at a tertiary tumor hospital between January 2003 and October 2013. METHODS: Baseline data of patients with primary hepatocellular carcinoma treated between January 2003 and October 2013 were retrospectively collected. Tumor stage was determined according to the Barcelona Clinic Liver Cancer (BCLC) staging system and Hong Kong Clinic Liver Cancer (HKLC) staging system. RESULTS: A total of 6241 patients with primary hepatocellular carcinoma were included in the analysis. In accordance with the BCLC, 28.9% of patients were in stage 0/A, 16.2% in stage B, 53.6% in stage C, and 1.3% in stage D. According to the HKLC stage system, 8.4% patients were in stage I, 1.5% in stage IIa, 29.0% in stage IIb, 10.0% in stage IIIa, 33.6% in stage IIIb, 3.4% in stage IVa, 2.5% in stage IVb, 0.2% in stage Va, and 11.4% in stage Vb. Treatment modalities applied to this patient group were as follows: 33.3% of patients underwent hepatic resection, 36.7% underwent transarterial chemoembolization (TACE), 2.2% underwent radiotherapy, 0.9% underwent local ablated therapy, 8.8% underwent systemic chemotherapy, 4.2% underwent traditional herbal medicine therapy, 0.1% underwent targeted drug therapy, and 13.8% received no treatment. Hepatic resection was the most frequent therapy for patients with BCLC 0/A/B disease, and TACE was the initial therapy for patients with BCLC C disease. In the Hong Kong Clinic Liver Cancer staging system, the main treatments for HKLC I to IIIb disease is hepatic resection and TACE. Systemic chemotherapy was the initial therapy for patients with HKLC IVa/IVb disease. Most HKLC Va/Vb patients received traditional Chinese medicine treatment. CONCLUSION: Prevalence of stage BCLC B and C disease was high among our hepatocellular carcinoma patients. In Hong Kong Clinic Liver Cancer staging system, HKLC I to IIIb disease was high among our HCC patients. Hepatic resection and TACE are initial therapies.

Regulation of semaphorin 4D expression and cell proliferation of ovarian cancer by ERalpha and ERbeta

Ovarian cancer is one of the most common malignancies in women. Semaphorin 4D (sema 4D) is involved in the progress of multiple cancers. In the presence of estrogen-like ligands, estrogen receptors (ERα and ERβ) participate in the progress of breast and ovarian cancers by transcriptional regulation. The aim of the study was to investigate the role of sema 4D and elucidate the regulatory pattern of ERα and ERβ on sema 4D expression in ovarian cancers. Sema 4D levels were up-regulated in ovarian cancer SKOV-3 cells. Patients with malignant ovarian cancers had significantly higher sema 4D levels than controls, suggesting an oncogene role of sema 4D in ovarian cancer. ERα expressions were up-regulated in SKOV-3 cells compared with normal ovarian IOSE80 epithelial cells. Conversely, down-regulation of ERβ was observed in SKOV-3 cells. Forced over-expression of ERα and ERβ in SKOV-3 cells was manipulated to establish ERα+ and ERβ+ SKOV-3 cell lines. Incubation of ERα+ SKOV-3 cells with ERs agonist 17β-estradiol (E2) significantly enhanced sema 4D expression and rate of cell proliferation. Incubated with E2, ERβ+ SKOV-3 cells showed lower sema 4D expression and cell proliferation. Blocking ERα and ERβ activities with ICI182-780 inhibitor, sema 4D expressions and cell proliferation of ERα+ and ERβ+ SKOV-3 cells were recovered to control levels. Taken together, the data showed that sema 4D expression was positively correlated with the progress of ovarian cancer. ERα positively regulated sema 4D expression and accelerated cell proliferation. ERβ negatively regulated sema 4D expression and inhibited cell multiplication.

RESEARCH ARTICLE Molecular cloning and expression analysis of a matrix Gla protein gene in the spinyhead croaker, Collichthys lucidus.

The matrix Gla (gamma-carboxyglutamic acid-rich) protein (MGP), a vitamin K-dependent and Gla-containing protein, is a calcification inhibitor that mainly functions in tissue calcification and mineralization. In this study, we obtained the complete cDNA sequence of MGP from the spinyhead croaker (Collichthys lucidus), which we named Cl-MGP. Cl-MGP was 923 bp long with a 384-bp open reading fragment that encoded 127 amino acids. The predicted MGP protein sequence contained a 19-residue hydrophobic signal peptide, suggesting that it possesses secretory characteristics. The Gla domain and the invariant unit ErraEtCedyspC, which has been identified in all known vitamin K-dependent vertebrate proteins, were highly conserved in Cl-MGP, suggesting that it uses the same mechanism to function as the known proteins. An alignment analysis revealed that Cl-MGP had the highest identity with Larimichthys crocea (93%), which had lost five amino acid residues in the C-terminal. A quantitative real-time polymerase chain reaction revealed that Cl-MGP expression was highest in the gill, followed by the cholecyst and spleen, with almost no expression in the blood, muscle, or testes. The high Cl-MGP expression in the gill is similar to that observed in other fish species, but the relatively high expression found in the cholecyst and spleen is not seen in all species. Future studies should investigate the tissue distributions of both mRNA and proteins in different species, in order to understand the function and evolution of MGP in different species.

Role of ADH1B rs1229984 and ALDH2 rs671 gene polymorphisms in the development of Alzheimer's disease.

In the present study, we investigated the association between ADH1B rs1229984 and ALDH2 rs671 polymorphisms and the development of Alzheimer's disease in a Chinese population. Genotyping of the ADH1B rs1229984 and ALDH2 rs671 polymorphisms was carried out by polymerase chain reaction-restriction fragment length polymorphism. Logistic regression analyses revealed that the AA genotype of ADH1B rs1229984 was associated with an increased risk of Alzheimer's disease (OR = 2.54, 95%CI = 1.19-5.41). In addition, ADH1B rs1229984 was also associated with elevated risk of Alzheimer's disease in both dominant (OR = 1.78, 95%CI = 1.09-2.93) and recessive (OR = 2.33, 95%CI = 1.18-4.57) models. For ALDH2 rs671, the AA genotype was correlated with an increased risk of Alzheimer's disease as compared to the GG genotype (OR = 4.57, 95%CI = 1.60-14.01). The ALDH2 rs671 polymorphism was associated with Alzheimer's in both dominant (OR = 1.79, 95%CI = 1.08-2.97) and recessive (OR = 4.17, 95%CI = 1.49-12.67) models. In conclusion, we observed that ADH1B rs1229984 and ALDH2 rs671 polymorphisms increased the risk of Alzheimer's disease in all the genetic models.

Characterization of the complete mitochondrial genome of Portunus pelagicus with implications for phylogenomics.

This study determined the mitochondrial genome structure of the blue swimming crab (Portunus pelagicus), and elucidated its phylogenetic relationships among the species within the order Decapoda. The complete mitochondrial genome was 16,155 bp long, and contained 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and 1 DNA control region. The gene order of the genome was the same as that found within the family Portunidae. Twenty-three genes were on the heavy strand and 14 were on the light strand. Almost all of the protein-coding genes were initiated by an ATG codon, except for three genes (ATP6, ND1, and ND3) that started with a rare ATT codon. Of the 13 protein-coding genes, 10 ended with complete TAA or TAG stop codons and three ended with an incomplete T codon. Thirteen non-coding regions were identified that ranged from 1 to 30 bp in length. Nine overlaps were found, which ranged 1 to 7 bp in length. Phylogenetic analyses based on 12 concatenated protein-coding genes revealed that P. pelagicus formed a monophyletic group with Portunus trituberculatus, which were in a larger group with Callinectes sapidus, while the genera Charybdis and Thalamita formed another group. These two groups clustered together and grouped with the genus Scylla. The phylogenetic analysis supported the inclusion of Charybdis in subfamily Portuninae of the family Portunidae, and revealed a close relationship between Charybdis and Thalamita. We suggest that Thalamita should also be classified into the subfamily Portuninae. The results can be used in the study of phylogenetic, population genetic and conservation genetics of P. pelagicus.

Optimization of selective breeding through analysis of morphological traits in Chinese sea bass (Lateolabrax maculatus).

Determining correlations between certain traits of economic importance constitutes an essential component of selective activities. In this study, our aim was to provide effective indicators for breeding programs of Lateolabrax maculatus, an important aquaculture species in China. We analyzed correlations between 20 morphometric traits and body weight, using correlation and path analyses. The results indicated that the correlations among all 21 traits were highly significant, with the highest correlation coefficient identified between total length and body weight. The path analysis indicated that total length (X1), body width (X5), distance from first dorsal fin origin to anal fin origin (X10), snout length (X16), eye diameter (X17), eye cross (X18), and slanting distance from snout tip to first dorsal fin origin (X19) significantly affected body weight (Y) directly. The following multiple-regression equation was obtained using stepwise multiple-regression analysis: Y = -472.108 + 1.065X1 + 7.728X5 + 1.973X10 - 7.024X16 - 4.400X17 - 3.338X18 + 2.138X19, with an adjusted multiple-correlation coefficient of 0.947. Body width had the largest determinant coefficient, as well as the highest positive direct correlation with body weight. At the same time, high indirect effects with six other morphometric traits on L. maculatus body weight, through body width, were identified. Hence, body width could be a key factor that efficiently indicates significant effects on body weight in L. maculatus.

Characterization of the complete mitochondrial genome and phylogenetic relationships of the three-spot swimming crab (Portunus sanguinolentus).

In this study, we determined the whole mitochondrial genome profile of the three-spot swimming crab (Portunus sanguinolentus) and elucidated phylogenetic relationships between representative species in the order Decapoda. The mitochondrial genome was 16,024 bp in length and consisted of 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a putative control region. Of the 37 genes, 23 were encoded by the heavy strand while 14 were encoded by the light strand. Four types of start codons were identified; ATG initiated nine genes, ATT initiated two genes, and ATC and GTG each started one gene. Nine protein-coding genes ended with a complete TAA or TAG stop codon, and four genes ended with an incomplete T or TA codon. Fourteen non-coding regions were found, which ranged from 1 to 34 bp in length. Nine overlaps were observed, with lengths between 1 and 7 bp. Phylogenetic analysis suggested that P. sanguinolentus is genetically closest to P. trituberculatus and P. pelagicus. Charybdis feriata, C. japonica, and Thalamita crenata formed a single cluster, and were close to the genera Callinectes and Portunus. Therefore, the genera Charybdis and Thalamita should be classified into the subfamily Portuninae.

Effect of uric acid on mitochondrial function and oxidative stress in hepatocytes.

Here, we investigated the effect of uric acid (UA) on hepatocyte mitochondria. Hepatocytes cultured in vitro were treated with varying concentrations of UA. The change in apoptotic activity was detected by flow cytometry. The DNA damage index 8-hydroxy-deoxy-guanosine (8-OHdG) and mitochondrial function indices succinate dehydrogenase (SDH), cytochrome C oxidase (CCO), and adenosine triphosphate (ATP) were detected by enzyme assays. Reactive oxygen species (ROS) accumulation was confirmed by a dichloro-dihydro-fluorescein diacetate assay. We observed an increase in apoptotic activity, ROS accumulation, and 8-OHdG activity in hepatocytes treated with UA for extended periods, indicating DNA damage; specifically, we observed a significant increase in these activities 48, 72, and 96 h after UA addition, compared to those observed at 24 h (P < 0.05). Cells treated with 30 mg/dL UA for 96 h showed a peak in apoptotic activity. We also observed a significant decrease in ATP, SDH, and CCO activities with the increase in uric acid concentration over time. Cells treated with 30 mg/dL UA for 96 h showed the highest ATP levels, while SDH and CCO activities at 48, 72, and 96 h post-UA treatment were significantly lower than those at 24 h (P < 0.01). Moreover, cells treated with 30 mg/dL UA showed a 0.02 ± 0.02 and 0.15 ± 0.01 mmol/ mg/min decrease in SDH and CCO levels after 72 h. Therefore, we concluded that high concentrations of UA may induce oxidative stress in hepatocyte mitochondria, increasing ROS production and ultimately resulting in mitochondrial damage.

Inhibitory effect of microRNA-27b on interleukin 17 (IL-17)-induced monocyte chemoattractant protein-1 (MCP1) expression.

We investigated the effect of microRNA-27b (miR-27b), a gene expression regulatory factor, on the expression of monocyte chemoattractant protein-1 (MCP1) stimulated by interleukin 17 (IL-17). After IL-17 had been added to H9C2 cardiomyocytes, an miR-27b mimic was transfected into the H9C2 cells. The mRNA expression levels of miR-27b and MCP1 in the H9C2 cells were detected by SYBR green I fluorescence quantitative reverse transcription polymerase chain reaction. Enzyme-linked immunosorbent assay was used to measure the expression levels of MCP1 protein in the H9C2 cells. The expression of MCP1 mRNA in the H9C2 cells began to increase 2 h after IL-17 stimulation, reached a peak at 4 h, and then decreased. The MCP1 protein level increased gradually in the 24 h following IL-17 stimulation. After transfection with the miR-27b mimic, the expression of miR-27b in the H9C2 cells significantly increased than that in the miRNA negative control group (P < 0.01). The MCP1 mRNA and protein levels in the miR-27b mimic + IL-17 group were significantly reduced than that in the miRNA negative control + IL-17 group (P < 0.01). miR-27b inhibited IL-17-induced MCP1 expression in the H9C2 cells, which demonstrates that treatment with microRNA could alleviate myocardial injury in viral myocarditis.

First genetic linkage map for the mud crab (Scylla paramamosain) constructed using microsatellite and AFLP markers.

The mud crab (Scylla paramamosain) is of economic importance for the fisheries and aquaculture industry in China. In this study, we constructed the first genetic linkage map for this species using microsatellite and amplified fragment length polymorphism (AFLP) markers. The map consisted of 65 linkage groups, including 34 triplets and 9 doublets. A total of 212 molecular markers were mapped, including 60 microsatellites and 152 AFLP markers. The linkage groups ranged from 7 to 102.5 cM and covered 2746.4 cM in length. The mean length was 42.3 cM per linkage group, and the mean spacing was 18.68 cM. The genome size was estimated to be 5539.62 cM, with 50% coverage by the present map. Moreover, we reported 5 transcriptome-derived polymorphic microsatellite markers and characterized their polymorphism in a first-generation family. This study will facilitate studies on high-density maps and molecular marker-assisted selection in S. paramamosain and related crustacean species.