Diagnostic performance of contrast-enhanced ultrasound (CEUS) combined with Ovarian-Adnexal Reporting and Data System (O-RADS) ultrasound risk stratification for adnexal masses: a systematic review and meta-analysis.

AIM: A number of studies have reported that contrast-enhanced ultrasound (CEUS) imaging might be used for the early diagnosis of adnexal masses. A meta-analysis was performed to evaluate the diagnostic accuracy of CEUS combined with Ovarian-Adnexal Reporting and Data System (O-RADS) ultrasound risk stratification for adnexal masses. MATERIALS AND METHODS: Related articles were retrieved from PubMed, Web of Science, Embase, and the Cochrane Library in strict accordance with established standards, and data (including true positive, false positive, false negative, and true negative values) was extracted from the original articles. The Quality Assessment of Diagnostic Accuracy Studies 2 was used to evaluate the quality of articles and the possibility of bias. STATA 12.0 software was used to perform statistical analysis. RESULTS: Five articles that included 598 patients were analyzed in this meta-analysis. The pooled sensitivity and specificity of CEUS combined with O-RADS for the diagnosis of adnexal masses were 0.95 (95% confidence interval [CI]: 0.91-0.98) and 0.86 (95% CI: 0.79-0.91). Moreover, the positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio (DOR), and area under the curve (AUC) were 6.81 (95% CI: 4.61-10.08), 0.05 (95% CI: 0.03-0.11), 111.30 (95% CI: 65.32-189.65), and 0.97 (95% CI: 0.95-0.98), respectively. The pooled AUC and DOR for the detection of CEUS combined with O-RADS were superior to O-RADS US. CONCLUSION: Our findings revealed that O-RADS combined with CEUS can improve the diagnostic accuracy of ovarian adnexal masses.

Qiliqiangxin capsule improves cardiac remodeling in rats with DOCA-salt-induced diastolic dysfunction.

OBJECTIVE: The aim of this study was to investigate the protective effect and mechanism of action (MOA) of Qiliqiangxin capsule (QL) in the deoxycorticosterone acetate (DOCA) salt-induced rat heart failure with preserved ejection fraction (HFpEF) model. MATERIALS AND METHODS: Nono-nephrectomy sixty Sprague Dawley (SD) rats received DOCA salt injection and 1% saline in drinking water for 4 weeks and were randomly divided into four groups on average: Model group (n=15), Sac/Val group (Sacubitril Valsartan 0.02 g/kg, n=15), QL-L group (Qiliqiangxin 0.25 g/kg, n=15) and QL-H group (Qiliqiangxin 1 g/kg, n=15). Another Normal group was set (n=15). Blood pressure, N-terminal pro-brain natriuretic peptide (NT-proBNP), cardiac index, echocardiography, and hemodynamics were measured to evaluate heart function. Masson and Wheat germ agglutinin (WGA) staining was performed to observe the fibrosis deposition and the cross-sectional area (CSA) of cardiomyocytes. The concentration levels of the serum cytokines, including tumor necrosis factor-α (TNF-α), interleukin (IL)-2, IL-6, and IL-10 inflammatory factors, were detected by ELISA; matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9), transforming growth factor-ß1 (TGF-ß1), nuclear factor-κB (NF-κB), Smad homologue 2 (Smad2) and Smad homologue 3 (Smad3) expression were detected by Western-blot. RESULTS: Compared with the Model group, QL treatment significantly ameliorated the heart function in DOCA salt-induced rat HFpEF model, showing a decrease in cardiac index, an increase of the EF and E/A ratio, a reduction in the left ventricular anterior/posterior wall (LVAW/LVPW), in the time contraction of isovolumic diastolic time (IVRT), -dP/dt Max, and Tau, and the decrease of serum NT-ProBNP. Masson and WGA staining indicated that QL inhibited the fibrosis deposition and the myocardial hypertrophy compared with the Model group, which was consistent in reducing the protein expression levels of cardiac remodeling such as TGF-ß1, MMP2, MMP9, Smad2, and Smad3. Moreover, QL treatment inhibited the expression of NF-κB in the heart tissues and decreased the serum concentration of pro-inflammatory cytokines TNF-α and IL-2, instead, increasing the IL-10 concentration. CONCLUSIONS: QL improved the cardiac function and inhibited the myocardial fibrosis in DOCA salt-induced rat HFpEF by improving diastolic dysfunction, preventing left ventricular hypertrophy, and ameliorating the inflammatory responses model in DOCA salt-induced rat HFpEF model.

Effects of individualized antiplatelet therapy based on CYP2C19 genotype and platelet function on the prognosis of patients after PCI.

OBJECTIVE: To evaluate the effect of individualized antiplatelet therapy based on CYP2C19 genotype and platelet function on the prognosis of patients after percutaneous coronary intervention (PCI) compared with conventional antiplatelet therapy. PATIENTS AND METHODS: Patients diagnosed with acute coronary syndromes (ACS) in Shandong Provincial Qianfoshan Hospital from December 2014 to December 2017 were included in this prospective study and randomly divided into conventional (CA) and individualized antiplatelet therapy group (IA) at 1:1 ratio. Patients in the CA group received clopidogrel 75 mg once a day (QD). Group IA was divided into extensive, intermediate, and poor metabolizers according to the results of the CYP2C19 gene test. Three genotypes were given clopidogrel 75 mg QD, 75 mg twice daily (BID) and ticagrelor 90 mg BID respectively. After taking these medicines for a period of time, platelet function was monitored by thromboelastography (TEG) and MAADP values were recorded. MAADP indicates the adenosine diphosphate (ADP) induced platelet function that not inhibited by medicine. High platelet reactivity (HPR) was defined as MAADP > 47mm, indicating a high risk of thrombus, and MAADP ≤ 31 mm indicates a high risk of hemorrhage. For extensive metabolizers (EMs) and intermediate metabolizers (IMs) patients with HPR, the antiplatelet therapy would be changed by the clinician according to the patient's conditions. Major adverse cardiovascular events (MACE) and hemorrhage events were monitored during 1-year follow-up. RESULTS: The patients with MAADP > 47 mm were 89 (28.6%) in the IA group. There were 50 EMs patients with MAADP > 47 mm (33.3%). Of which, there were 2 cases which changed the dosage of clopidogrel to 75 mg BID, 14 cases who changed clopidogrel to ticagrelor. There were 36 IMs patients with MAADP > 47 mm (30.8%). Of which, there were 19 cases who changed clopidogrel to ticagrelor. There was no significant difference in the value of MAADP between EMs and IMs patients. Within 1 year after PCI, the occurrence of MACE in the IA group was significantly lower than that in the CA group (p=0.010). CONCLUSIONS: (1) Patients with a CYP2C19 loss-of-function (LOF) gene who take double doses of clopidogrel overcome the decreased efficacy of clopidogrel which partly due to CYP2C19 LOF gene, without increasing the risk of hemorrhage. (2) Individualized antiplatelet therapy based on CYP2C19 genotype and platelet function can significantly reduce the occurrence of MACE (mainly acute non-fatal myocardial infarction) after PCI without increasing the risk of moderate or severe hemorrhage.

[Clinicopathological analysis of 98 cases of benign lymphoidenosis of oral mucosa].

Objective: To investigate the clinical manifestations and pathological changes of benign lymphoadenosis of oral mucosa. Methods: The clinical data of 98 cases of benign lymphoadenosis of oral mucosa were analyzed. Results: The clinical manifestations of benign lymphoadenosis of oral mucosa included erosive ulcer (64%) and nodule (9%) and the rate of misdiagnosis was 98%. Neutrophil infiltration occurred in the epithelium of 51% cases and the lymphocyte was diffusely infiltrated in lamina propria of 83% cases. Conclusions: When the mucous membrane of the lamina propria is characterized by complex cell components, diffuse infiltrating lymphocytes and infiltration of neutrophils in mucosal epithelium without erosion and ulceration, it is necessary to highly suspect benign lymphoadenosis of oral mucosa. Finding the focal aggregation of lymphoid follicles or lymphocytes is helpful for the correct diagnosis.

Sinus tarsi approach (STA) versus extensile lateral approach (ELA) for treatment of closed displaced intra-articular calcaneal fractures (DIACF): A meta-analysis.

INTRODUCTION: Our aim was to compare the effect of sinus tarsi approach (STA) vs extensile lateral approach (ELA) for treatment of closed displaced intra-articular calcaneal fractures (DIACF) is still being debated. MATERIALS AND METHODS: A thorough research was carried out in the MEDLINE, EMBASE and Cochrane library databases from inception to December 2016. Only prospective or retrospective comparative studies were selected in this meta-analysis. Two independent reviewers conducted literature search, data extraction and quality assessment. The primary outcomes were anatomical restoration and prevalence of complications. Secondary outcomes included operation time and functional recovery. RESULTS: Four randomized controlled trials involving 326 patients and three cohort studies involving 206 patients were included. STA technique for DIACFs led to a decline in both operation time and incidence of complications. There were no significant differences between the groups in American Orthopedic Foot and Ankle Society scores, nor changes in Böhler angle. CONCLUSIONS: This meta-analysis suggests that STA technique may reduce the operation time and incidence of complications. In conclusion, STA technique is reasonably an optimal choice for DIACF.

[Diagnostic value of albumin-bilirubin grade combined with serum ammonia in cirrhosis with hepatic encephalopathy].

Objective: To explore the value of albumin-bilirubin (ALBI) grade combined with serum ammonia in the diagnosis of cirrhosis with hepatic encephalopathy (HE). Methods: The serum level of total bilirubin(TBIL), albumin( ALB )and blood ammonia were detected in 139 patients including 73 cirrhosis patients without HE and 66 cirrhosis patients with HE from January 2015 to January 2017 in Beijing You'an Hospital, and the relationship between ALBI and blood ammonia value and Child grade and hepatic encephalopathy was analyzed. Results: The level of ALBI and blood ammonia were more and more higher with the increase of Child grade, the level of ALBI in Child A, B and C were -2.3±0.6, -1.7±0.5, -0.9±0.4, and there was a statistically significant(F=125.100, P<0.001). The blood ammonia concentration in Child A, B and C were(42.6±16.0), (56.1±31.2), (69.8±34.7) µmol/L, and there was a statistically significant(F=7.400, P<0.001). The level of ALBI was higher with the increase of model for end-stage liver disease (MELD) grade, and there was a positive correlation(r=0.547, P<0.001). The ALBI value in the HE group was higher than the cirrhosis patients without HE((-1.1±0.5)vs(-1.6±0.7)), and the difference was statistically significant (t=5.244, P<0.001). Level of blood ammonia in the HE group was(83.6±39.5)µmol/L, which was higher than the level of cirrhosis patients without HE(42.9±17.0)µmol/L, and the difference was statistically significant (t=8.130, P<0.001) . When ALBI and blood ammonia were combined, the ROC curve area was 0.911, the sensitivity was 93.9%, the specificity was 93.2%. Conclusion: There is a significant diagnosis value and high clinical application when ALBI is combined with blood ammonia to diagnose HE .

[Naked cuticle homolog 2 positively regulates the osteogenic differentiation of rat dental follicle cells].

Objective: To examine the expression of naked cuticle homolog 2 (Nkd2) in the process of root development and osteogenic differentiation of dental follicle cells of rat (rDFC), in order to explore the molecular mechanisms of Nkd2 on the osteoblast differentiation of rDFCs. Methods: Immunohistochemical analysis was used to detect the expression of Nkd2 in the base dental follicle of the mandibular first molar of rat at 1, 3, 5, 7, 9, 11 and 13 days postnatal. Mineralization nodule formation of rDFCs was detected by alizarin red staining and cetylpyridine. The change of Nkd2 during osteogenic differentiation of rDFCs was evaluated by Western blotting and the associations between Nkd2 and osteogenic cytokines of alkaline phosphatase (ALP), Runt-related transcription factor-2 (RUNX2) and osteocalcin (OCN) were examined. The rDFCs were transfected with small interfering RNA (siRNA) to knock down the expression of Nkd2 and Western blotting and quantitative real-time PCR (qPCR) were adopted to explore the effects of Nkd2 on osteogenic differentiation by detecting variations of Nkd2 and osteogenic factors ALP, RUNX2, OCN among silencing group (Si), negative control RNA group (Nc) and mock control group (Mock), respectively. Results: The expression of Nkd2 in the base dental follicle of the mandibular first molar of rat was time dependent. Mineralization nodules of rDFCs and absorbance of cetylpyridine after osteogenic induction increased gradually (the absorbances of cetylpyridine were 0 week: 0.017±0.005, 1 week: 0.702±0.044, 2 weeks: 1.812±0.531, 3 weeks: 2.767±0.253, respectively). Results of Western blotting showed that Nkd2 (1.60±0.23) of mineralization group was significantly higher than that of control group (1) (P<0.05) at the early stage of osteogenic differentiation along with the expression of other osteogenic factors. The protein and mRNA of Nkd2 and osteogenic factors were significantly decreased in Si group compared with Nc and Mock groups (P<0.05), and no changes between Nc and Mock groups were observed. The changes of protein in Si, Nc and Mock groups were Nkd2: 0.42±0.10, 1.12±0.07, 1, ALP: 0.70±0.15, 1.11±0.14, 1, RUNX2: 0.58±0.08, 0.93±0.08, 1 and OCN: 0.64±0.06, 0.99±0.02, 1, respectively. The mRNA variances in Si, Nc and Mock groups were Nkd2: 0.39±0.05, 0.96±0.10, 1, ALP: 0.15±0.13, 1.01±0.07, 1, RUNX2: 0.39±0.31, 0.97±0.13, 1, OCN: 0.17±0.08, 1.08±0.21, 1, respectively. Conclusions: Nkd2 participates in the root development process in rat and may acts as a positive role in the early stage of osteogenic differentiation of rDFCs in rat.

Molecular cloning, overexpression, purification, and sequence analysis of the giant panda (Ailuropoda melanoleuca) ferritin light polypeptide.

The complementary DNA (cDNA) of the giant panda (Ailuropoda melanoleuca) ferritin light polypeptide (FTL) gene was successfully cloned using reverse transcription-polymerase chain reaction technology. We constructed a recombinant expression vector containing FTL cDNA and overexpressed it in Escherichia coli using pET28a plasmids. The expressed protein was then purified by nickel chelate affinity chromatography. The cloned cDNA fragment was 580 bp long and contained an open reading frame of 525 bp. The deduced protein sequence was composed of 175 amino acids and had an estimated molecular weight of 19.90 kDa, with an isoelectric point of 5.53. Topology prediction revealed one N-glycosylation site, two casein kinase II phosphorylation sites, one N-myristoylation site, two protein kinase C phosphorylation sites, and one cell attachment sequence. Alignment indicated that the nucleotide and deduced amino acid sequences are highly conserved across several mammals, including Homo sapiens, Cavia porcellus, Equus caballus, and Felis catus, among others. The FTL gene was readily expressed in E. coli, which gave rise to the accumulation of a polypeptide of the expected size (25.50 kDa, including an N-terminal polyhistidine tag).

Comparative analysis and molecular characterization of genomic sequences and proteins of FABP4 and FABP5 from the giant panda (Ailuropoda melanoleuca).

Fatty acid binding proteins (FABPs) are a family of small, highly conserved cytoplasmic proteins that bind long-chain fatty acids and other hydrophobic ligands. In this study, cDNA and genomic sequences of FABP4 and FABP5 were cloned successfully from the giant panda (Ailuropoda melanoleuca) using reverse transcription polymerase chain reaction (RT-PCR) technology and touchdown-PCR. The cDNAs of FABP4 and FABP5 cloned from the giant panda were 400 and 413 bp in length, containing an open reading frame of 399 and 408 bp, encoding 132 and 135 amino acids, respectively. The genomic sequences of FABP4 and FABP5 were 3976 and 3962 bp, respectively, which each contained four exons and three introns. Sequence alignment indicated a high degree of homology with reported FABP sequences of other mammals at both the amino acid and DNA levels. Topology prediction revealed seven protein kinase C phosphorylation sites, two casein kinase II phosphorylation sites, two N-myristoylation sites, and one cytosolic fatty acid-binding protein signature in the FABP4 protein, and three N-glycosylation sites, three protein kinase C phosphorylation sites, one casein kinase II phosphorylation site, one N-myristoylation site, one amidation site, and one cytosolic fatty acid-binding protein signature in the FABP5 protein. The FABP4 and FABP5 genes were overexpressed in Escherichia coli BL21 and they produced the expected 16.8- and 17.0-kDa polypeptides. The results obtained in this study provide information for further in-depth research of this system, which has great value of both theoretical and practical significance.

Overexpression, purification, and pharmacologic evaluation of anticancer activity of ribosomal protein L24 from the giant panda (Ailuropoda melanoleuca).

The ribosomal protein L24 (RPL24) belongs to the L24E family of ribosomal proteins and is located in the cytoplasm. The purpose of this study was to investigate the structure and anti-cancer function of RPL24 of the giant panda (Ailuropoda melanoleuca). The complementary DNA of RPL24 was cloned successfully using reverse transcription-polymerase chain reaction technology. We constructed a recombinant expression vector containing RPL24 complementary DNA and overexpressed it in Escherichia coli using pET28a plasmids. The expression product obtained was purified using Ni-chelating affinity chromatography. The results indicated that the length of the fragment cloned is 509 bp, and it contains an open-reading frame of 474 bp encoding 157 amino acids. Primary structure analysis revealed that the molecular weight of the putative RPL24 protein is 17.78 kDa with a theoretical isoelectric point of 11.86. The RPL24 gene is readily expressed in E. coli, and the RPL24 fused with the N-terminal histidine-tagged protein to give rise to the accumulation of an expected 23.51-kDa polypeptide. The inhibitory rate in mice treated with 0.1 mg/mL RPL24, the highest of 3 doses administered, can reach 67.662%, which may be comparable to the response to mannatide. The histology of organs with tumors showed that the tissues in the RPL24 group displayed a looser arrangement compared with that in the control group. Furthermore, no obvious damage was apparent in other organs, such as heart, lung, and kidney. The data showed that the recombinant RPL24 had time and dose dependency on the cell growth inhibition rate. Human laryngeal carcinoma Hep-2 cells treated with 0.3125-10 µg/mL RPL24 for 24 h displayed significant cell growth inhibition (P < 0.05; N = 6) in assays using 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide compared with that in control (untreated) cells. By contrast, human hepatoma Hep G-2 cells displayed no significant change (P > 0.05; N = 6) from control (untreated) cells. RPL24 has time and dose dependency on Hep-2 cell growth inhibition. The data indicate that the effect at low concentrations is better than that at high concentrations, and the concentration of 0.625 µg/mL provides the best rate of growth inhibition. Further research is ongoing to determine the bioactive principles of recombinant RPL24 protein that are responsible for its anticancer activity.

CT and MRI in target delineation in primary hepatocellular carcinoma.

PURPOSE: To compare the quality of computed tomography (CT) and magnetic resonance imaging (MRI) in target delineation of hepatocellular carcinoma. PATIENTS AND METHODS: Thirty-one patients with hepatocellular carcinoma received CT and MRI (including diffusion-weighted imaging [DWI]) before surgery. The maximal measurement on axial imaging and pathologic examination were extracted for statistical analysis. RESULTS: CT, MRI, DWI-based tumour size correlated well with pathologic sizes, r=0.974, 0.969, 0.964 respectively. But Wilcoxon signed-ranked test showed that differences did exist. CT overestimated the tumour size by 2.9mm compared to pathology (95% CI: -13.2 to 7.4mm). The agreement of MRI-pathology seemed to be worse than CT, with a mean difference of 3.6mm (95% CI: -14.7 to 7.7mm). The worst agreement was in DWI-pathology, with a mean difference of 5mm (95% CI: -17.9 to 7.9mm). But significant difference was found neither between CT and MRI (P=0.477) nor between MRI and DWI (P=0.079). CONCLUSIONS: CT and MRI-based tumour size correlated well with pathologic size, but differences did exist. Most of the lesions were overestimated by CT and MRI. CT and MRI were similar in the guidance of target delineation, and DWI had added little value to MRI. A margin of 10mm around the gross tumour volume to become the clinical target volume is likely not sufficient.

cDNA, genomic sequence cloning and overexpression of giant panda (Ailuropoda melanoleuca) mitochondrial ATP synthase ATP5G1.

The ATP5G1 gene is one of the three genes that encode mitochondrial ATP synthase subunit c of the proton channel. We cloned the cDNA and determined the genomic sequence of the ATP5G1 gene from the giant panda (Ailuropoda melanoleuca) using RT-PCR technology and touchdown-PCR, respectively. The cloned cDNA fragment contains an open reading frame of 411 bp encoding 136 amino acids; the length of the genomic sequence is of 1838 bp, containing three exons and two introns. Alignment analysis revealed that the nucleotide sequence and the deduced protein sequence are highly conserved compared to Homo sapiens, Mus musculus, Rattus norvegicus, Bos taurus, and Sus scrofa. The homologies for nucleotide sequences of the giant panda ATP5G1 to those of these species are 93.92, 92.21, 92.46, 93.67, and 92.46%, respectively, and the homologies for amino acid sequences are 90.44, 95.59, 93.38, 94.12, and 91.91%, respectively. Topology prediction showed that there is one protein kinase C phosphorylation site, one casein kinase II phosphorylation site, five N-myristoylation sites, and one ATP synthase c subunit signature in the ATP5G1 protein of the giant panda. The cDNA of ATP5G1 was transfected into Escherichia coli, and the ATP5G1 fused with the N-terminally GST-tagged protein gave rise to accumulation of an expected 40-kDa polypeptide, which had the characteristics of the predicted protein.

cDNA, genomic sequence cloning and overexpression of ribosomal protein gene L9 (rpL9) of the giant panda (Ailuropoda melanoleuca).

The ribosomal protein L9 (RPL9), a component of the large subunit of the ribosome, has an unusual structure, comprising two compact globular domains connected by an α-helix; it interacts with 23 S rRNA. To obtain information about rpL9 of Ailuropoda melanoleuca (the giant panda) we designed primers based on the known mammalian nucleotide sequence. RT-PCR and PCR strategies were employed to isolate cDNA and the rpL9 gene from A. melanoleuca; these were sequenced and analyzed. We overexpressed cDNA of the rpL9 gene in Escherichia coli BL21. The cloned cDNA fragment was 627 bp in length, containing an open reading frame of 579 bp. The deduced protein is composed of 192 amino acids, with an estimated molecular mass of 21.86 kDa and an isoelectric point of 10.36. The length of the genomic sequence is 3807 bp, including six exons and five introns. Based on alignment analysis, rpL9 has high similarity among species; we found 85% agreement of DNA and amino acid sequences with the other species that have been analyzed. Based on topology predictions, there are two N-glycosylation sites, five protein kinase C phosphorylation sites, one casein kinase II phosphorylation site, two tyrosine kinase phosphorylation sites, three N-myristoylation sites, one amidation site, and one ribosomal protein L6 signature 2 in the L9 protein of A. melanoleuca. The rpL9 gene can be readily expressed in E. coli; it fuses with the N-terminal GST-tagged protein, giving rise to the accumulation of an expected 26.51-kDa polypeptide, which is in good agreement with the predicted molecular weight. This expression product could be used for purification and further study of its function.

Polyphenols from Parabarium huaitingii and their positive inotropic and anti-myocardial infarction effects in rats.

Eight phenolic compounds, including (-)-epicatechin (1) and seven proanthocyanidins (2-8), were obtained from the butanol extract of Parabarium huaitingii (PHB). Their chemical structures were identified based on analyses of mass spectra (MS), NMR, CD spectra, and partial acid catalyzed thiolytic degradation. The observation made by laser scanning confocal microscope found a significant increase of the concentration of intracellular Ca²+ ([Ca²+](i)) in single myocytes when the PHB was added, while compounds 1 and 3 had the same physiological effect. Further investigations showed PHB had a dose-dependent positive inotropic effect on isolated right atria and papillary muscle of left ventricle of the rat, while having no significant influence on the spontaneous beating rate of the isolated right atria. The inotropic effect of PHB could be greatly abolished by pretreating the myocardium in Ca²+-free solution. These findings indicated that PHB could significantly increase [Ca²+](i) in myocytes, which was greatly dependent on the influx of extracellular Ca²+. Compounds 1 and 3 might be the effective ingredients of the inotropic effect of PHB. In addition, PHB could also significantly decrease the infarct size of the heart on acute myocardial infarction (AMI) model rats, which suggested its myocardial protective effect on ischemic myocardium. The positive inotropic effect of PHB, together with its myocardial protective effect on AMI, suggested that PHB had a promising potential for the prevention and treatment of heart failure, especially the one that was caused by AMI.

cDNA, genomic sequence cloning and overexpression of the ribosomal protein S13 gene in the giant panda (Ailuropoda melanoleuca).

The cDNA and the genomic sequence of ribosomal protein S13 (RPS13) of the giant panda (Ailuropoda melanoleuca) was cloned using reverse transcription-polymerase chain reaction (RT-PCR) and touchdown-PCR, respectively. These two sequences were sequenced and analyzed, and the cDNA of the RPS13 gene was overexpressed in Escherichia coli BL21. We compared the nucleotide sequences of the coding region and the amino acid sequences with those of seven other mammalian species retrieved from GenBank. The cDNA fragment of the RPS13 cloned from the giant panda is 496 bp in size, containing an open-reading frame of 456 bp, encoding 151 amino acids. The length of the genomic sequence is 2277 bp, with five exons and four introns. The coding sequence shows a high degree of homology to those of Homo sapiens, Bos taurus, Canis lupus familiaris, Macaca mulatta, Mus musculus, Rattus norvegicus, and Pan troglodytes; the degree of homology was 91.23, 94.30, 94.74, 92.11, 87.94, 87.72, and 91.45%, respectively. The homologies for the deduced amino acid sequences reached as high as 99%. Primary structure analysis revealed that the molecular weight of the putative RPS13 protein is 17.22325 kDa, with a theoretical pI of 10.42. Based on topology prediction, there is one protein kinase C phosphorylation site, one casein kinase II phosphorylation site, two N-myristoylation sites, and one ribosomal protein S15 signature in the RPS13 protein of the giant panda. The RPS13 gene can be expressed in E. coli and the RPS13 protein fused with the N-terminally GST-tagged form, which gave rise to the addition of an expected 43-kDa polypeptide.

Giant panda ribosomal protein S14: cDNA, genomic sequence cloning, sequence analysis, and overexpression.

RPS14 is a component of the 40S ribosomal subunit encoded by the RPS14 gene and is required for its maturation. The cDNA and the genomic sequence of RPS14 were cloned successfully from the giant panda (Ailuropoda melanoleuca) using RT-PCR technology and touchdown-PCR, respectively; they were both sequenced and analyzed. The length of the cloned cDNA fragment was 492 bp; it contained an open-reading frame of 456 bp, encoding 151 amino acids. The length of the genomic sequence is 3421 bp; it contains four exons and three introns. Alignment analysis indicates that the nucleotide sequence shares a high degree of homology with those of Homo sapiens, Bos taurus, Mus musculus, Rattus norvegicus, Gallus gallus, Xenopus laevis, and Danio rerio (93.64, 83.37, 92.54, 91.89, 87.28, 84.21, and 84.87%, respectively). Comparison of the deduced amino acid sequences of the giant panda with those of these other species revealed that the RPS14 of giant panda is highly homologous with those of B. taurus, R. norvegicus and D. rerio (85.99, 99.34 and 99.34%, respectively), and is 100% identical with the others. This degree of conservation of RPS14 suggests evolutionary selection. Topology prediction shows that there are two N-glycosylation sites, three protein kinase C phosphorylation sites, two casein kinase II phosphorylation sites, four N-myristoylation sites, two amidation sites, and one ribosomal protein S11 signature in the RPS14 protein of the giant panda. The RPS14 gene can be readily expressed in Escherichia coli. When it was fused with the N-terminally His-tagged protein, it gave rise to accumulation of an expected 22-kDa polypeptide, in good agreement with the predicted molecular weight. The expression product obtained can be purified for studies of its function.

The role of renin-angiotensin system blockade therapy in the prevention of atrial fibrillation: a meta-analysis of randomized controlled trials.

Current evidence suggests that angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin II receptor blockers (ARBs) have emerged as novel drugs for preventing the development of atrial fibrillation (AF). A meta-analysis was performed of 26 randomized controlled clinical trials evaluating the effect of ACEIs or ARBs on the prevention of AF. Overall, ACEIs and ARBs revealed statistically significant preventive effects on AF (odds ratio (OR), 0.65; 95% confidence interval (CI), 0.55-0.76). The preventive effect was similar in the two classes of drugs (ACEI: OR, 0.68; ARB: OR, 0.69). ACEIs and ARBs showed greater preventive effects on recurrent AF (OR, 0.45; 95% CI, 0.31-0.65) than on new-onset AF (OR, 0.80; 95% CI, 0.70-0.92). Prevention was greatest in patients with AF who were receiving amiodarone as a basic treatment drug (OR, 0.35; 95% CI, 0.26-0.48). In patients with heart failure, there appeared to be a large effect (OR, 0.497), but the credible interval (CrI) limits were wide (95% CrI, 0.187-1.161).

Monitoring of CaSO(4) deposition behaviors on biofilm during nanofiltration via ultrasonic time-domain reflectometry (UTDR).

The ultrasonic time-domain reflectometry (UTDR) as a non-destructive real-time method was employed to monitor the CaSO(4) deposition behaviors on biofilm during nanofiltration (NF). Two parallel experiments were performed to compare the different behaviors of CaSO(4) deposition with and without biofilm on the membrane. Results showed that the flux decline during combined fouling was slower than that in case of CaSO(4) fouling alone. The Ca(2 + ) rejection obtained with biofilm was higher than that without. A larger acoustic differential signal obtained by UTDR in the combined fouling revealed a denser and thicker layer formed on the membrane surface. Furthermore, the amount of CaSO(4) deposition on the biofouled membrane was more than that on non-biofouled membrane as a result of microorganisms as crystal nucleus to induce CaSO(4) crystallization and deposition. SEM images indicate that the CaSO(4) crystals deposited in order on the non-biofouled membrane, whereas on the biofouled membrane they were embedded in the biofilm. The denser and thicker fouling layer formed with biofilm was impermeable, resulting in a high Ca(2 + ) rejection. The complexation of Ca with polysaccharide in biofilm would eliminate the cake-enhanced osmotic pressure effect leading to a slow flux decline. To sum up, the independent measurements corroborate the ultrasonic measurements.

Effects of the sample size of reference population on determining BMD reference curve and peak BMD and diagnosing osteoporosis.

UNLABELLED: Establishing reference databases generally requires a large sample size to achieve reliable results. Our study revealed that the varying sample size from hundreds to thousands of individuals has no decisive effect on the bone mineral density (BMD) reference curve, peak BMD, and diagnosing osteoporosis. It provides a reference point for determining the sample size while establishing local BMD reference databases. INTRODUCTION: This study attempts to determine a suitable sample size for establishing bone mineral density (BMD) reference databases in a local laboratory. METHODS: The total reference population consisted of 3,662 Chinese females aged 6-85 years. BMDs were measured with a dual-energy X-ray absorptiometry densitometer. The subjects were randomly divided into four different sample groups, that is, total number (Tn) = 3,662, 1/2n = 1,831, 1/4n = 916, and 1/8n = 458. We used the best regression model to determine BMD reference curve and peak BMD. RESULTS: There was no significant difference in the full curves between the four sample groups at each skeletal site, although some discrepancy at the end of the curves was observed at the spine. Peak BMDs were very similar in the four sample groups. According to the Chinese diagnostic criteria (BMD >25% below the peak BMD as osteoporosis), no difference was observed in the osteoporosis detection rate using the reference values determined by the four different sample groups. CONCLUSIONS: Varying the sample size from hundreds to thousands has no decisive effect on establishing BMD reference curve and determining peak BMD. It should be practical for determining the reference population while establishing local BMD databases.