Potential role for the germ cell-specific Rad21 in early meiosis of oocyte and spermatocyte in the Chinese mitten crab Eriocheir sinensis.

Rad21/Rec8 family proteins are vital for sister chromatid segregation in mitosis and homologous recombination in meiosis, but no molecular data are available in crustacean species. In this study, a germ cell-specific Rad21 named EsRad21 was identified in the crab Eriocheir sinensis. EsRad21 mRNA has an open reading frame of 2310 base pairs (bp) encoding a 769 amino acids (aa) protein. RT-PCR showed that EsRad21 mRNA was particularly expressed in testis and ovary. The RT-qPCR results further revealed that the EsRad21 mRNA exhibited similar expression pattern in gonads at various developmental stages. EsRad21 mRNA expression level was the highest in testis at early spermatogenesis stage and ovaries at previtellogenesis stage, thereafter decreased significantly at middle spermatogenesis and vitellogenesis, and finally reach the lowest level at late spermatogenesis and vitellogenesis. In situ hybridization (ISH) analysis showed that EsRad21 mRNA was exclusively expressed in germline cells, but not in gonadal somatic cells. Notably, hybridized signal was detected on chromosomes of metaphase spermatocytes. EsRad21 is thus an underlying helpful indicator of the early phases of germ cell development. RNAi knockdown of EsRad21 downregulated the expression of other meiosis-related genes like Smc5-Smc6 and SPO11 and resulted in high mortality of individuals after 24 h post injection of EsRad21 dsRNA. Taken together, our results showed a potential role for EsRad21 in early meiosis of oocytes and spermatocytes in E. sinensis. This is the first report on the molecular characterization of the Rad21 transcript in a crustacean species.

Chemical constituents from the stems of Acanthopanax senticosus with their inhibitory activity on α-glucosidase.

A new aurone named (2Z)-2-[(4'-hydroxy-3'-methoxyphenyl) methylene]-6-methoxy-7-prenyl-3(2H)-benzofurane (1), together with five known compounds (2-6), were isolated from EtOAc-soluble extract of the stems of Acanthopanax senticosus. The chemical structures were elucidated on the basis of spectroscopic analyses. All isolates were evaluated for in vitro inhibitory activity on α-glucosidase. Among them, compounds 1 and 4 were found to exhibit moderate inhibitory activity on α-glucosidase with IC50 value of 64.1 ± 1.2 and 48.9 ± 1.1 µM, respectively.[Formula: see text].

Emerging of a novel natural recombinant fowl adenovirus in China.

Outbreaks of hydropericardium syndrome and inclusion body hepatitis caused by fowl adenovirus (FAdV) have occurred in China since June 2015, resulting in significant economic loss to poultry industry. In this study, a novel FAdV, designated as AH720, with recombination among serotype FAdV-8a and FAdV-8b was isolated and characterized in China. Full genome analysis revealed that the AH720 has the genome backbone from FAdV-8b and the fibre gene from FAdV-8a. In an infection study, although AH720 was not lethal to chickens, AH720 did cause characteristic lesions of inclusion body hepatitis in the infected chickens. All these data not only provide strong evidences for the recombination among different serotype FAdVs, but also highlight the necessary for monitoring the molecular epidemiology of such recombinant FAdV to develop efficient strategies against FAdV.

Effect of clinical pharmacist intervention on the treatment of acute pancreatitis.

Background The participation of clinical pharmacists in the treatment of acute pancreatitis has rarely been reported. Objective The aim of this study was to retrospectively evaluate the impact of intervention of clinical pharmacists on the treatment of acute pancreatitis. Setting An academic teaching hospital in Taizhou, Jiangsu, China. Method Two hundred and twenty-eight patients with acute pancreatitis were retrospectively enrolled from July 2017 to July 2018 and divided into an intervention group (n = 119) and a control group (n = 109) according to whether a clinical pharmacist was involved. No significant differences in the baseline clinical characteristics were found between the groups. Clinical pharmacists participated in drug formulation and adjustment, pharmaceutical care, and follow-up. Main outcome measure Clinical outcomes, average hospital stays, costs, incidence of adverse drug reactions, 1-month subsequent visit rate, and patient satisfaction between the two groups were measured. Results The clinical symptoms of patients in both groups were relieved after treatment. There were no significant differences between the groups in computed tomography grades after treatment, incidence of adverse drug reactions, or average hospital stays. However, the intervention group had lower total costs of hospitalization, drugs and antibiotics but higher rates of 1-month subsequent visits and satisfaction compared with the control group. Conclusion The intervention of clinical pharmacists in the treatment of acute pancreatitis can effectively reduce costs of hospitalization, drug and antibiotics and improve follow-up compliance and patient satisfaction.

Salmonella in Retail Food and Wild Birds in Singapore-Prevalence, Antimicrobial Resistance, and Sequence Types.

Non-typhoidal salmonellosis is a leading cause of foodborne zoonosis. To better understand the epidemiology of human salmonellosis, this study aimed to determine the prevalence, antimicrobial resistance and sequence types of Salmonella in retail food and wild birds (proximity to humans) in Singapore. We analyzed 21,428 cooked and ready-to-eat food and 1,510 residual faecal samples of wild birds collected during 2010-2015. Thirty-two Salmonella isolates from food and wild birds were subjected to disc diffusion and multi-locus sequence typing (MLST). Salmonella was isolated from 0.08% (17/21,428) of food and 0.99% (15/1510) of wild birds. None of the isolates from wild birds (n = 15) exhibited phenotypic resistance, while the isolates from food (47.1%, 8/17) showed a high prevalence of phenotypic resistance to, at least, one antimicrobial. These findings suggested that the avian Salmonella isolates had been subjected to less antimicrobial selection pressure than those from food samples. MLST revealed specific sequence types found in both food and wild birds. The study can guide future studies with whole-genome analysis on a larger number of isolates from various sectors for public health measures.

Actein ameliorates hepatic steatosis and fibrosis in high fat diet-induced NAFLD by regulation of insulin and leptin resistant.

Insulin and leptin resistance are highly involved in metabolic syndrome and non-alcoholic fatty liver disease (NAFLD). Presently, no approved treatment is available. Actein is isolated from the rthizomes of Cimicifuga foetida, a triterpene glycoside, exhibiting important biological properties, such as anti-inflammatory, anti-cancer, and anti-oxidant activity. However, its effects on metabolic syndrome are poorly understood. The aims of the study were mainly to investigate the molecular mechanisms regulating insulin and leptin resistance, and lipogenic action of actein in high fat diet-fed mice. Our data indicated that actein-treated mice displayed lower body weight, epididymal and subcutaneous fat mass, as well as serum lipid levels. Also, improved insulin and leptin resistance were observed in actein-treated groups. Liver inflammation and fibrosis triggered by high fat diet were decreased for actein administration. Moreover, hepatic lipid accumulation was also reduced by actein along with reductions of hepatic de novo lipogenesis-linked signals in actein-treated rodents with high fat diet. High fat diet-induced activation of insulin receptor substrate 1/Forkhead box protein O1 (IRS1/FOXO1), Janus kinase 2 gene/signal transducer and activator of transcription (JAK2/STAT3) and Protein Kinase B/Glycogen synthase kinase 3 beta (AKT/GSK3ß) pathways in liver was inhibited by actein, a potential mechanism by which hyperinsulinemia, hyperleptindemia and dyslipidemia were attenuated. Thus, the findings above might be of nutritional and therapeutic importance for the treatment of NAFLD.

Mitochondrial genome of the sucking disc gudgeon, Discogobio yunnanensis (Teleostei, Cypriniformes).

The complete mitochondrial genome of Discogobio yunnanensis was determined using the polymerase chain reaction and directly sequenced with the primer walking method. The whole mitogenome was 16,602 bp in length, consisting of 13 protein-coding genes, 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (rRNA) genes and 1 control region. It had the typical circular molecule structure of vertebrate's mitogenome. The whole base composition was estimated to be 32.10% A, 26.79% T, 25.95% C, 15.16% G with AT bias of 58.89%. The complete mitogenome of D. yunnanensis provides the fundamental data for conservation genetics study on this freshwater fish species.

[Effects of nitrogen application and winter green manure on soil active organic carbon and the soil carbon pool management index].

Based on a cropping system of "winter green manure-double rice", the 4 x 4 two-factor test was used to study the effects of different nitrogen (N) application levels and winter green manure application on soil active organic carbon (AOC) and the C pool management index. The aim was to explore the ecological effects of winter green manure on soil improvement and determine the appropriate application levels of N fertilizer and winter green manure for improved rice yield. Results were as follows: 1) Compared with the control, the SOC and AOC contents increased by 22.2% and 26.7%, respectively, under the green manure only treatment, but the SOC contents decreased by 0.6%-3.4% under the single N fertilizer treatment. Compared with the control, the soil C pool management index increased by 24.55 and 15.17 under the green manure only and green manure plus N fertilizer treatments, respectively, and reduced by 2.59 under the single N fertilizer treatment. Compared with no fertilization, the average microbial biomass carbon (MBC) increased by 54.0%, 95.2% and 14.3% under the green manure, green manure plus N fertilizer and single N fertilizer treatments, respectively. 2) The soil AOC content was significantly positively correlated with the C pool management index (P < 0.01), and had a significant correlation with dis- solved organic C and MBC (P < 0.05). Rice yield was significantly positively correlated with AOC contents and the C pool management index, and the correlation coefficient was significantly greater than that with the total organic C. These results suggested that application of winter green manure at proper rates with inorganic fertilizer could increase SOC contents and the soil C pool management index, improve soil quality and fertility.

Molecularly imprinted electrochemical sensor based on amine group modified graphene covalently linked electrode for 4-nonylphenol detection.

In this work, an imprinted electrochemical sensor based on electrochemical reduced graphene covalently modified carbon electrode was developed for the determination of 4-nonylphenol (NP). An amine-terminated functional graphene oxide was covalently modified onto the electrode surface with diazonium salt reactions to improve the stability and reproducibility of the imprinted sensor. The electrochemical properties of each modified electrodes were investigated with differential pulse voltammetry (DPV). The electrochemical characteristic of the imprinted sensor was also investigated using electrochemical impedance spectroscopy (EIS) in detail. The response currents of the imprinted electrode exhibited a linear relationship toward 4-nonylphenol concentration ranging from 1.0 × 10(-11) to 1.0 × 10(-8) gm L(-1) with the detection limit of 3.5 × 10(-12) gm L(-1) (S/N=3). The fabricated electrochemical imprinted sensor was successfully applied to the detection of 4-nonylphenol in rain and lake water samples.

Molecularly imprinted electrochemical sensor based on a reduced graphene modified carbon electrode for tetrabromobisphenol A detection.

A simple strategy for the indirect detection of 3,3',5,5'-tetrabromobisphenol A (TBBPA) was developed with a surface imprinted sensor based on an electrochemical reduced graphene modified carbon electrode. The preparation procedure of imprinted electrode and the response mechanism of the imprinted electrode toward TBBPA are discussed in detail. The electrochemical characteristics of the imprinted sensor were investigated using cyclic voltammetry and their morphologies were characterized using scanning electron microscopy. The indirect detection for TBBPA was successfully implemented by monitoring the peak current of Fe(CN)6(3-/4-)-TBBPA complex. The response currents of the imprinted sensor exhibited a linear relationship toward the TBBPA concentration range from 0.5 to 4.5 nM with a detection limit of 0.23 nM (S/N = 3). The fabricated electrochemical imprinted sensor was successfully applied to the detection of TBBPA in rain and lake water samples using the standard addition method. With the advantages of high sensitivity and simple operation, the determination methodology is a promising candidate for TBBPA detection in real water samples.

[Surgical treatment of thoracic ossification of ligamentum flavum and therapeutic effects analysis].

OBJECTIVE: To investigate the surgical treatment of thoracic ossification of the ligamentum flavum (TOLF) and analysis of the therapeutic effects. METHODS: Six patients with thoracic ossification of the ligamentum flavum were retrospectively studied from October 2006 to October 2009. All of the patients in this group were treated by en bloc hemi-articular process laminectomy. There were 4 males and 2 females, ranging in age from 45 to 66 years, averaged 55.2 years. The evaluate factors including the preoperative and postoperative JOA scores,and the function of the urinary bladder were analyzed. RESULTS: All the 6 patients were followed after operation. The mean followed-up duration was 10.5 months, ranging from 2 to 18 months. According to JOA evaluation criteria, 4 patients got an excellent result, 1 good and 1 bad. The one patient with bad result had spinal cord compressed for too long time, and the T2-weight MRI showed nonreversible degeneration of spinal cord and combined with schizophrenia. The clinical symptoms improved with varying degrees. CONCLUSION: The en bloc hemi-articular process laminectomy is an effective method for ossification of the thoracic ligamentum flavum. A thorough decompression and real-time protection of the spinal cord is the key to the success of surgery.

[Preparation of monoclonal antibodies against HN protein of Class I Newcastle disease virus virulent strain].

AIM: To prepare monoclonal antibodies (mAb) against HN protein of Class I Newcaslte disease virus. METHODS: Several 6-week-old mice were immunized with ClassI 9a5b virulent strain. Hemagglutination inhibition (HI), indirect immunofluorescence assay (IFA), cell neutralization test, ELISA and Western blot analysis were used to characterize and identify these antibodies. RESULTS: Four hybridoma cell lines were successfully prepared, designated as mAb 2F5, 2F12, 3H7 and 3H9. Among them, only mAb 3H7 and 3H9 recognized HN protein of the ClassI strain, specifically neutralizing NDV and inhibiting hemagglutination. CONCLUSION: mAb 3H7 and 3H9 could be of use in identification of Class I and Class II strains, and in functional studies of HN and cell receptors for NDV.

[Complete genomic sequence of a watermelon isolate of cucumber green mottle mosaic virus in northern China].

The complete genomic sequence of a watermelon isolate of Cucumber green mottle mosaic virus (CGMMV-LN) in Liaoning province was determined and compared with other cucurbit-infecting tobamoviruses. The genomic RNA of CGMMV-LN comprised 6422 nt, and 5'- and 3'- noncoding regions consisted of 59 nt and 175 nt, respectively. The encoded four proteins were two replicase proteins of 186 kD and 129 kD, move protein of 29 kD and coat protein of 17.4 kD. The alignment results of complete nucleotide sequence showed that CGMMV-LN shared identities of 97.6%-99.3% with four other CGMMV isolates, but only shared identities of 61.7%-62.8% with three other tobamoviruses. Homology trees generated from replicase proteins of 186 kD and coat proteins suggested that cucurbit-infecting tobamoviruses could be separated into two subgroups: subgroup I comprising all the isolates of CGMMV and subgroup II comprising Cucumber fruit mottle mosaic virus, Kyuri green mottle mosaic virus and Zucchini green mottle mosaic virus.

[Preparation and immunological characterization of monoclonal antibodies against VP22 carboxyl terminus of Marek's disease virus serotype 1].

AIM: To prepare and characterize monoclonal antibodies (mAb) against VP22 carboxyl terminus of CVI988/Rispens strain of Marek's disease virus serotype 1. METHODS: Carboxyl terminus of CVI988 VP22 (94aa-243aa) was expressed in prokaryotic system. mAbs against VP22 were prepared by hybridoma technology from BALB/c mice immunized with the fusion protein GST-VP22C and characterized by ELISA, indirect fluorescence assay (IFA) and Western blot. RESULTS: Two hybridoma cell lines stably secreting mAb against VP22C were obtained and designated as 3F7 and 4E4. mAb 3F7 could react with VP22 expressed in all the plaques, while mAb 4E4 stained all the cells nuclei in MDV-infected CEF cells. It was also found that 3F7 could react with VP22 expressed in Sf9 cells and denatured VP22 by Western blot analysis. In addition, it was further showed that the epitope of mAb 3F7 was located within the domain between 94aa and 193aa, the predicted site of protein transduction domain of VP22. CONCLUSION: The preparation of the mAb is very important to further research in protein transduction domain of MDV-1 VP22.

In situ extraction of intracellular L-asparaginase using thermoseparating aqueous two-phase systems.

The feasibility and generic applicability of directly integrating conventional discrete operations of cell disruption by high pressure homogenizer and the product capture by aqueous two-phase extraction (ATPE) system have been demonstrated for the extraction of intracellular L-asparaginase from E. coli. In a side-by-side comparison with the conventional ATPE process, including cell disruption, centrifugal clarification and following ATPE, purification of L-asparaginase via this novel in situ ATPE process yielded a product of L-asparaginase with a higher specific activity of 94.8 U/(mg protein) and a higher yield of 73.3%, both of which in the conventional ATPE process were 78.6 U/(mg protein) and 52.1%, respectively. In the purification of L-asparaginase (pI=4.9), product-debris interactions commonly diminish its recovery. It was demonstrated that immediate extraction of L-asparaginase in ATPE systems when it is released at pH 5.0 during cell disruption effectively increased its recovery in the top phase due to the reduced interaction between L-asparaginase and cell debris and the reduced degradation by contaminated protease. In addition, no clarification step and/or disruptate storage are required in this in situ ATPE, which reduced the number of unit operations and thus shortened the overall process time. This novel process has a good potential for the separation of other intracellular biological products.

[Construction and immunological characterization of recombinant Marek's disease virus expressing IBDV VP2 fusion protein].

A transfer plasmid vector pUC18-US10-VP2 was first constructed by inserting the gene of the enhancer green fluorescent protein(eGFP) fused to the VP2 gene of very virulent Infectious bursal disease virus (IBDV) JS strain into the US10 fragment of the Marek's disease virus (MDV) CV1988/Rispens. The recombinant virus, designated as rMDV, was developed by co-transfecting CEF with the transfer plasmid vector and simultaneously infecting with the CVI988/Rispens virus. The PCR and IFA results indicated that the rMDV is stable after 31 passages. Chickens vaccinated with rMDV were protected from challenge with 100LD50 of IBDV JS. The protection ratio of the chickens vaccinated with the 1000PFU, 2000PFU, 5000PFU of the rMDV were 50%, 60%, and 80% respectively. It is interesting that the average histopathology BF lesion scores of chicken group immunized with 5000PFU of rMDV by one-time vaccination was close to that of chicken group vaccinated with IBDV live vaccine NF8 strain for twice (2.0/1.5). There is no difference in protection between the groups (P > 0.05) but significent difference between groups immunized with 5000 PFU of rMDV and with normal MDV. This demonstrated that rMDV expressing VP2 fusion protein was effective vaccine against IBDV in SPF chickens.

Effects of enhanced counterpulsation on vascular cell release of coagulation factors.

BACKGROUND: Enhanced external counterpulsation (EECP), a noninvasive treatment for patients with angina pectoris, provides long-term benefits of decreased anginal frequency and improved exercise tolerance. Previous studies have suggested that shear stress may result in angiogenesis and alter endothelial hemostatic factor release. Whether EECP therapy effects an alteration in endothelial cell proliferation and function remains unclear. The level of vascular endothelial growth factor (VEGF) and four other endothelial hemostatic factors (tissue plasminogen activator, plasminogen activator inhibitor-1, von Willebrand factor, and D-dimers) were measured in patients before and after 35 hours of EECP treatment. METHODS: Plasma levels of endothelial growth and hemostatic factors were assessed using the standard enzyme-linked immunosorbent method. RESULTS: No significant difference in the hemostatic factors and VEGF after EECP treatment was revealed; there was a trend toward an increase in VEGF levels posttreatment. CONCLUSIONS: Vascular endothelial cells play a critical role in the regulation of coagulation because they control the expression of tissue plasminogen activator, plasminogen activator inhibitor-1, von Willebrand factor, and D-dimers. Our results suggest EECP may not play a role in controlling coagulation in patients with coronary artery disease through release of endothelial hemostatic factors. Although there was a tendency for increased VEGF release, larger studies are necessary to confirm these observations.

Interleukin-2 levels are associated with carotid artery intima-media thickness.

OBJECTIVE: To show that serum interleukin levels are associated with carotid intima-media thickness (IMT). BACKGROUND: Inflammation is hypothesized to play a central role in atherogenesis, and serum markers of inflammation are predictive of cardiovascular disease. Interleukin-2, a pro-inflammatory cytokine produced largely by naive CD4 T cells and Th1 (pro-inflammatory) T cells, has been found in a high proportion of carotid plaques. METHODS: High-resolution ultrasound of the carotid arteries and serum cytokine levels were measured in stroke-free participants. The mean of the maximum IMT in bilateral bifurcation, common and internal carotid artery segments was measured. Serum levels of interleukin (IL)-1beta, IL-2, IL-6, C-reactive protein, tumor necrosis factor (TNF) alpha and TNF receptors were measured using enzyme-linked immunosorbent assay. RESULTS: IL-2 levels were significantly correlated with IMT (r=0.33, P<0.0001), but other cytokines were not. Each unit increase in IL-2 was significantly associated with a mean increase in IMT of 0.18 mm (P=0.0001). After adjusting for other atherosclerotic risk factors, the association was unchanged (mean increase in IMT per unit increase IL-2=0.18 mm, P<0.0001). Each standard deviation increase in the level of IL-2 was associated with an increased risk (adjusted odds ratio 1.80, 95% CI 1.12-2.89) for an IMT> or =1.0mm (75th percentile for IMT). CONCLUSION: Serum levels of IL-2, a pro-inflammatory cytokine, are associated with carotid artery IMT, a predictor of stroke and vascular disease. Serum inflammatory markers may provide a novel marker of atherosclerotic risk, and inflammation may provide a new therapeutic target for stroke prevention.

Dynamic variability of hemostatic and fibrinolytic factors in young women.

This prospective study was designed to characterize the time course and variability of hemostatic and fibrinolytic risk factors over the course of a menstrual cycle in normal premenopausal women. Plasminogen activator inhibitor (PAI-1), tissue plasminogen activator, von Willebrand factor, fibrinogen, and fibrin D-dimer predict risk of coronary heart disease. Yet there is limited information describing the status of endogenous hormone concentrations and hemostatic and coagulation factors in premenopausal women. Twenty premenopausal women, mean age 34 +/- 7 yr, underwent testing over a cycle to measure endogenous hormones and hemostatic factors: estradiol and progesterone, FSH, LH; PAI-1, tissue plasminogen activator, von Willebrand factor, fibrin D-dimer, and fibrinogen as well as lipids: total cholesterol, high-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol, and triglycerides. There was cyclical variability in estradiol (P < 0.01) and progesterone (P < 0.001) during the follicular and luteal phases. Moreover, there was intra- and interindividual cyclical variation in hemostatic risk factors. Measures of PAI-1 (P < 0.01) and D-dimer (P < 0.05) differed during the follicular and luteal phases. As estradiol concentration increased, PAI-I decreased. There was a significant correlation between total cholesterol and PAI-1 (r = 0.56, P < 0.05), low-density lipoprotein-cholesterol and PAI-1 (r = 0.50, P < 0.05) as well as between total cholesterol and fibrinogen (r = 0.61, P < 0.05). There is significant cyclical variability in estradiol, FSH, and progesterone as well as the hemostatic factors, PAI-1 and fibrin D-dimer. Characterization of emerging hemostatic risk factors enhances understanding of normal physiology, provides insight into the relation between estrogen and hemostatic factors, and raises the potential for predicting coronary heart disease even in relatively young women.

[FTIR investigation of new polymer solid electrolytes].

The conductivity of the porous polymer solid electrolyte blended with PVDF and PMMA, which was made by a micro-wave hot-cross-linking method, reached 2.05 x 10(-3) S x cm(-1) at room temperature. The polymer solid electrolyte was analyzed and investigated by FTIR. The results show that the PVDF, PMMA and LiClO4 in the polymer solid electrolyte were not simply blended, but certain kind of effect existed which was strengthened only when the polymer solid electrolyte came into being.