Potential Clinical Values Research Articles

The characterization, renoprotection and antioxidation of enzymatic and acidic exopolysaccharides from Hypsizigus marmoreus

The present work was designed to investigate the characterization, as well as the antioxidation and renoprotection in streptozocin (STZ)-induced diabetic mice, of exopolysaccharides (EPS) and the enzymatic-EPS (EEPS) and acidic-EPS (AEPS) hydrolysates, which were separated from the fermentation liquor of Hypsizigus marmoreus. Animal results demonstrated that EPS, EEPS and AEPS had potential antioxidant and renoprotective effects, especially EEPS. Additionally, they were the most effective, reflecting increases in superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), total antioxidant capacity (T-AOC), and albumin (ALB) of 168.33%, 124.8%, 268.17% 179.49%, and 68.71%, respectively, and decreases in the contents of malondialdehyde (MDA), lipid peroxide (LPO) and levels of serum urea nitrogen (BUN) and creatinine (CRE) by 70.58%, 58.43%, 23.97% and 29.60%, respectively, at a dose of 800 mg/kg compared to those of model mice. Three polysaccharides ameliorated the histopathological alterations which were observed in the kidney of diabetic mice. Furthermore, the characterization of polysaccharides had been expressed. These findings indicated that the EEPS from H. marmoreus possesses more effective renoprotection and antioxidation effects and provided insight into its potential clinical values on preventing diabetes.

Optical coherence tomography angiography of acute Vogt-Koyanagi-Harada disease

Objective: To measure vascular density in retinal and choroidal capillary layers by optical coherence tomography angiography (OCTA) and to explore their potential clinical values in Vogt-Koyanagi-Harada disease (VKH). Methods: This is a cross-sectional case-control study. Twenty-one acute VKH cases presented to Peking Union Medical College Hospital between April 2015 and July 2015 and 49 healthy controls were enrolled in this study. OCTA was performed with a split-spectrum amplitude-decorrelation angiography-optical coherence tomography (SSADA-OCT) system borrowed from Optovue Inc. Vascular densities of the capillary layers were analyzed by the built-in software. Bilateral independent sample t-test was used to compare retinal and choroidal vascular density of eyes in acute phase VKH with healthy controls. Spearman correlation test was used to analyze the correlation between the vascular density of retina and choroid in acute phase VKH and LogMAR best corrected visual acuity (BCVA). Results: Mean vascular density of superficial capillary plexus (SCP), deep capillary plexus (DCP) and choriocapillaris (CC) in eyes of the acute VKH group were statistically lower than those of healthy control group (51.56%±2.88%, 56.51%±2.31%, 58.14%±2.10% vs. 53.55%±2.51%, 58.14%±2.10%, 58.14%±2.10%, t=-3.890, 3.910,-6.554, all 3 P values were<0.01). In VKH patients, eyes with retinal detachment (RD) had statistically lower mean vascular density in the CC layer than those without (NRD) (63.46%±2.19% vs. 65.85%±1.79%, t=3.890, P<0.01), while no statistically significant differences were found in vascular densities of SCP and DCP layers between RD and NRD eyes. Spearman's correlation test revealed a negative correlation between vascular density of the CC layer and logMAR BCVA(r=-0.437, P<0.01), while no statistically significant associations were found between logMAR BCVA and vascular densities of SCP and DCP. Conclusions: OCTA can be used for quantitative detection of vascular density of retina and choroidal capillary in acute VKH. There is circulatory disturbance in SCP, DCP and CC of acute VKH. Vascular density of CC is associated with visual acuity and retinal detachment. (Chin J Ophthalmol, 2017, 53: 735-739).

Long non-coding RNAs in osteosarcoma.

Long non-coding RNAs (lncRNAs) with more than 200 nuleotides, have been explored to participate in various cancer types including osteosarcoma (OS), which is the most common kind of primary bone tumors with high morbidity in infants and adolescents. These oncogenic or tumor suppressive lncRNAs regulate OS pathogenesis, such as cell growth, proliferation, invasion, migration, metastasis and cell apoptosis, serve as independent prognostic biomarkers or play a significant role in multidrug resistance (MDR) in OS cells. In this review, we attempt to dissect the participation of lncRNAs in pathogenesis of OS and their potential clinical values, and also provide an outlook for viable biomarkers and therapeutic targets in OS.

Role of AXL expression in non-small cell lung cancer.

The present study aimed to investigate the expression profile of AXL in non-small cell lung cancer (NSCLC) and its clinical significance. The current study included 257 NSCLC patients, tyrosine-protein kinase receptor UFO (AXL) expression in paired lung cancer and adjacent normal lung tissues of NSCLC patients were compared by immunohistochemistry, western blot analysis and quantitative polymerase chain reaction (qPCR). These methods were used to detect the expression of the AXL gene and protein in fresh tissues from 35 patients. Small interfering RNA (siRNA) was transfected into the H1299 lung cancer cell line to knock down AXL expression; the effects of AXL-siRNA on cell proliferation and migration were examined by MTT and Transwell migration assay, respectively. It was found that AXL staining density in lung cancer tissues was significantly increased compared with adjacent normal lung tissues (55.25 vs. 26.85%; P<0.01); and the expression level of AXL in NSCLC patients was significantly associated with the degree of tumor differentiation (P<0.01) and the clinical stage of disease (P<0.01). Western blotting and qPCR showed that AXL expression was significantly higher in cancer tissues compared with that in adjacent lung tissue (P<0.05). Additionally, the current study also showed that AXL-siRNA inhibited H1299 cell proliferation and migration in vitro. The present study demonstrates the association between increased expression of AXL in NSCLC and the low differentiation phenotype, and its effects on cell proliferation and migration, suggesting its potential clinical values for the prognosis of NSCLC.

MetaKTSP: a meta-analytic top scoring pair method for robust cross-study validation of omics prediction analysis.

Supervised machine learning is widely applied to transcriptomic data to predict disease diagnosis, prognosis or survival. Robust and interpretable classifiers with high accuracy are usually favored for their clinical and translational potential. The top scoring pair (TSP) algorithm is an example that applies a simple rank-based algorithm to identify rank-altered gene pairs for classifier construction. Although many classification methods perform well in cross-validation of single expression profile, the performance usually greatly reduces in cross-study validation (i.e. the prediction model is established in the training study and applied to an independent test study) for all machine learning methods, including TSP. The failure of cross-study validation has largely diminished the potential translational and clinical values of the models. The purpose of this article is to develop a meta-analytic top scoring pair (MetaKTSP) framework that combines multiple transcriptomic studies and generates a robust prediction model applicable to independent test studies. We proposed two frameworks, by averaging TSP scores or by combining P-values from individual studies, to select the top gene pairs for model construction. We applied the proposed methods in simulated data sets and three large-scale real applications in breast cancer, idiopathic pulmonary fibrosis and pan-cancer methylation. The result showed superior performance of cross-study validation accuracy and biomarker selection for the new meta-analytic framework. In conclusion, combining multiple omics data sets in the public domain increases robustness and accuracy of the classification model that will ultimately improve disease understanding and clinical treatment decisions to benefit patients. An R package MetaKTSP is available online. (http://tsenglab.biostat.pitt.edu/software.htm). ctseng@pitt.edu Supplementary data are available at Bioinformatics online.

Transcatheter arterial infusion chemotherapy increases expression level of miR-142-5p in stage III colorectal cancer.

To investigate the expression level of miR-142-5p and its potential target gene endothelial PAS domain protein 1(EPAS1) in Stage III colorectal cancer during Transcatheter arterial infusion chemotherapy (TAI). Illumina high-throughput sequencing was used to obtain miRNA expression profiles of paired tumor and adjacent normal tissues from one patient received TAI 1 week before the operation and another patient directly underwent an operation. The expression levels of miR-142-5p was measured with both high-throughput sequencing and quantitative real time-polymerase chain reaction. The expression levels of miR-142-5p, were significantly reduced in tumor tissues of stage III CRC, then significantly increased in tumor tissues receiving TAI and higher than tumor tissues without TAI. The apoptosis rate of HT-29 colon cancer cells was mildly increased after transfection with pre-miR-142. miR-142-5p could bind directly to the 3'untranslated region of endothelial PAS domain protein 1 and reduce its expression. miR-142-5p is a potential tumor suppressor in CRC and is upregulated in tumor tissues after TAI, suggesting its potential clinical values for testing the functionality of TAI and predicting the progress of CRC.

Application of lymphocyte-based in vitro tests to the diagnosis of drug eruption

T lymphocyte-mediated delayed type hypersensitivity plays a very important role in the pathogenesis of drug eruption.Currently,T lymphocyte-based in vitro tests have attracted more and more attention.Compared to skin tests,these tests show higher sensitivity with no potential danger to patients,and can evaluate cross-reactivity between similar drugs simultaneously.T lymphocyte-based in vitro tests,including lymphocyte transformation test,cytokine release assay,detection of cytotoxic components and cell activation markers,can be used for the etiological diagnosis of drug eruption and have potential clinical application values. Key words: Drug eruptions; Lymphocytes; Allergy; Diagnosis

QM-FISH analysis of the genes involved in the G1/S checkpoint signaling pathway in triple-negative breast cancer

This study was conducted to analyze copy number alterations (CNAs) of the genes involved in the G1/S checkpoint signaling pathway of triple-negative breast cancer (TNBC) and to evaluate their clinical value in the prognosis of TNBC. Quantitative multi-gene fluorescence in situ hybridization was used to study CNAs of the genes involved in the G1/S checkpoint signaling pathway, including cyclin d1 (CCND1), c-Myc, p21, cell-cycle-checkpoint kinase 2 gene, p16, retinoblastoma (Rb1), murine double minute 2 (Mdm2) and p53, in 60 TNBC samples and 60 non-TNBC samples. In comparison with the non-TNBC samples, CNAs of the genes involved in the G1/S checkpoint signaling pathway were more frequently observed in the TNBC samples (p = 0.000). Out of a total of eight genes, six (CCND1, c-Myc, p16, Rb1, Mdm2, and p53) exhibited significantly different CNAs between the TNBC group and the non-TNBC group. Univariate survival analysis revealed that the gene amplification of c-Myc (p = 0.008), Mdm2 (p = 0.020) and the gene deletion of p21 (p = 0.004), p16 (p = 0.015), and Rb1 (p = 0.028) were the independent predictive factor of 5-year OS for patients with TNBC. Cox multivariate analysis revealed that the gene amplification of c-Myc (p = 0.026) and the gene deletion of p21 (p = 0.019) and p16 (p = 0.034) were independent prognostic factors affecting the 5-year OS for TNBC. CNAs of the genes involved in the G1/S checkpoint signaling pathway presented a higher rate of incidence in TNBC than in non-TNBC, which could indicate one of the molecular mechanisms for the specific biological characteristics of TNBC. The genes c-Myc, p21, and p16 were correlated with the prognosis of TNBC and therefore may have potential clinical application values in the prognostic prediction of TNBC.

Expression and clinical significance of EphA8 in colon cancer

Objective To investigate the expression of EphA8 in colon cancer and its clinical significance.Methods Real-time quantitative polymerase chain reaction (RT-PCR) and Western blot were respectively used to assess the expressions of EphA8 mRNA and protein in normal colon mucosa cell line and colon cancer cell lines.Immunohistochemistry for EphA8 and vascular endothelial growth factor (VEGF) were performed in 98 cases of colon cancer and 12 matched normal mucosa tissues.CD31 immunohistochemical staining was used for microvascular density (MVD) counting.The relationships between the expression of EphA8 and the expression of VEGF and MVD,the clinical pathological significance,and the prognosis of patients were analyzed by statistical methods.Results EphA8 mRNA expressions were significantly higher in colon cancer cell lines than those in normal mucosa cell line (t =11.98,13.54,P ＜0.001).EphA8 protein expressions were significantly higher in colon cancer cell lines than those in normal mucosa cell line (t =4.63,P =0.006;t =4.92,P =0.004).The high expression of EphA8 was closely related to tumor size (x2 =22.97,P＜0.001),TNM stage (P＜0.001),differentiated degree (P =0.007)and lymph node metastasis (P＜0.001),distant metastasis (x2 =6.97,P =0.008) and poor survival rote (x2 =17.3,P ＜0.001).But it was not associated with the gender and age (x2 =1.36,P =0.30; x2 =0.83,P=0.44).Besides,EphA8 had positive correlation with the VEGF and MVD (r =0.434,P ＜0.001; r =0.584,P ＜0.001).Conclusion EphA8 may play important roles in the development and metastasis of colon cancer,which has potential clinical values in the therapeutic intervention and prognosis of colon cancer. Key words: Colonic neoplasms; Receptor, EphA8; Angiopoietins

Diffusion-weighted imaging (DWI) of the spleen in patients with liver cirrhosis and portal hypertension

ObjectiveThe purpose of this study was to assess the influence of liver cirrhosis and portal hypertension on diffusion coefficients of the spleen. Material and MethodsWe retrospectively evaluated 50 patients with liver cirrhosis and 50 patients without any history of liver disease who underwent magnetic resonance imaging of the upper abdomen, including echo planar diffusion-weighted imaging using b values of 50, 300 and 600mm2/s. Spleen apparent diffusion coefficient (ADC), liver ADC, muscle ADC and normalized spleen ADC (defined as the ratio of spleen ADC to muscle ADC) were compared between cirrhotic patients and patients in the control group and correlated with Child–Pugh stages. Reproducibility was assessed by measuring interclass correlation coefficient (n=11). Additionally, in eight patients, ADC measurements were performed 1 day before and 3 days after transjugular intrahepatic portosystemic shunt (TIPSS) implantation. ResultsCompared with control subjects, patients with cirrhosis and portal hypertension had significantly higher spleen ADCs (P=.0001). There was a statistically significant correlation between Child–Pugh grade and spleen ADC (Pearson correlation coefficient, observer 1 r=0.6, P=.0001; observer 2 r=0.5, P=.0001). After TIPSS implantation, we observed a reduction in spleen ADC values. Spleen ADC measurements showed a high reproducibility (interclass correlation coefficient 0.75, P=.001). ConclusionOur data suggest that different stages of liver cirrhosis and portal hypertension correlate with ADC values of the spleen. Furthermore, ADC values of the spleen decrease after TIPSS implantation. Further studies are required to understand the potential clinical values of these observations.

Quantitative analysis of circulating DNA in plasma of patients with acute pancreatitis and its potential clinical value

Objective To quantitatively detect the circulating DNA in plasma of patients with acute pancreatitis and to evaluate its potential clinical values. Methods Blood samples collected from 40 patients with mild acute pancreatitis (MAP), 20 with severe acute pancreatitis (SAP) and 50healthy controls were extracted for DNA using genomic DNA extraction kit. The DNA level was detected by real-time fluorescence quantitative PCR assay. The clinical association of DNA level with acute pancreatitis was analyzed. The receiver operating characteristic (ROC) curve was set up. Results The median concentration of plasma DNA in SAP group was 24.84 ng/ml, which was significantly higher than that in MAP group (7.60 ng/ml, P=0.006) and the healthy controls (5.23 ng/ml, P=0.000). The median concentration of plasma DNA in patients with acute pancreatitis was higher than in healthy controls (P=0.006), however, there was no significant difference between MAP group and healthy controls (P=0.322).The area under the ROC curve performed by the plasma DNA from SAP and MAP groups was 0. 881(95% CI, 0.773-0.989). With a cutoff value of 11.20 ng/ml, the overall sensitivity was 19/20% and specificity was 72.5%. Plasma DNA level was found to be associated with APACHE-Ⅱ score (P=0.001), Ranson score (P=0. 013), serum Ca2+ level (P=0.000), and c-reactive protein levels (P=0.001). Conclusions Plasma DNA is correlated with the extent of pancreatitis. It can be used in monitoring the development of acute pancreatitis and may be a potential marker for early diagnosis of SAP. Key words: Pancreatitis,acute necrotizing; Polymerase chain reaction; DNA

Clinical Audit in Gynecological Cancer Surgery: Development of a Risk Scoring System to Predict Adverse Events

Major risk factors in the general surgical population for morbidity and mortality have been identified using a number of risk scoring systems. Such risk factors include older age, length of surgery, anemia, and patients' comorbidities. A US Veteran's Affairs hospital program evaluated measures to reduce adverse events (AEs) and improve intra- and postoperative outcome variables in the over one million major operations occurring in Veteran's Affairs hospitals over a 10-year period (1991-2001). The data showed a 27% and 45% reduction in postoperative mortality and morbidity respectively. This program has been a model for developing a validated risk adjusted model for morbidity and mortality for different surgical specialties. There is no validated risk factor model for gynecological cancer. The aim of this study was to determine the incidence of AEs in patients with suspected or proven gynecological cancer, and to develop a clinical risk scoring system to predict the development of AEs. Between 2007 and 2008, 369 patients with suspected gynecological malignancies underwent a laparotomy or a laparoscopic procedure at a tertiary referral center for gynecological cancer. Surgical AEs were prospectively recorded and information was extracted from the medical records on potential patient risk factors, demographics, clinical values and histopathological findings. Multivariate logistic regression analysis was used to determine independent risk factors predictive for any surgical AE. The overall risk score (RS) for each variable was determined as follows: The coefficients from the multivariate model were scaled by a factor of 2; rounding the values obtained to the nearest integer provided the risk points. The sum of the risk points was the overall RS. Of the 369 patients, 95 (25.8%) developed at least one intra- and postoperative AE. Twenty-nine (8%) patients had intraoperative ARs and 77 (21%) experienced postoperative AEs; 11 patients (3.0%) developed both. Independent risk factors predictive of surgical AEs were surgical complexity, elevated SGOT (serum glutamic oxaloacetic transaminase, >=35 U/liter), elevated body mass index, and higher ASA (American Society of Anesthesiology) scores. The overall RS for an AR ranged from 0 to 14. The probability of having an AR was determined with the following formula: Risk (%) for an AR = 100/(1 + e(3.697 - (risk score/2)). These findings demonstrate that quantification of surgical risks in gynecological cancer surgery based on this model system may allow cross comparison of AE rates with other studies and has the potential to help reduce adverse events. Obesity is the only independent risk factor identified that is generally modifiable. Copyright (c) 2000-2010 Ovid Technologies, Inc.

Bacterial Strain-to-Strain Variation in Pharmacodynamic Index Magnitude, a Hitherto Unconsidered Factor in Establishing Antibiotic Clinical Breakpoints

Antibiotic pharmacodynamic modeling allows variations in pathogen susceptibility and human pharmacokinetics to be accounted for when considering antibiotic doses, potential bacterial pathogen targets for therapy, and clinical susceptibility breakpoints. Variation in the pharmacodynamic index (area-under-the-concentration curve to 24 h [AUC(24)]/MIC; maximum serum concentration of drug in the serum/MIC; time the serum concentration remains higher than the MIC [T > MIC]) is not usually considered. In an in vitro pharmacokinetic model of infection using a dose-ranging design, we established the relationship between AUC(24)/MIC and the antibacterial effect for moxifloxacin against 10 strains of Staphylococcus aureus. The distributions of AUC(24)/MIC targets for 24-h bacteriostatic effect and 1-log, 2-log, and 3-log drops in bacterial counts were used to calculate potential clinical breakpoint values, and these were compared with those obtained by the more conventional approach of taking a single AUC(24)/MIC target. Consideration of the AUC(24)/MIC as a distribution rather than a single value resulted in a lower clinical breakpoint.

Differential regulation of dehydroepiandrosterone and estrogen on bone and uterus in ovariectomized mice

Dehydroepiandrosterone (DHEA) may be useful in the treatment of postmenopausal osteoporosis (PMO). Our present study has found the preferable stimulatory effect of DHEA on bone, in contrast to the proliferative effects of estradiol (E2) on the endometrium and the uterus, which suggests that DHEA has greater potential clinical value than estrogens in prophylaxis and therapeutics for PMO. A series of findings raise the possibility that DHEA may be useful in the treatment of PMO. Our present study thus aimed at the differential effects of DHEA and E2 on bone and the uterus in ovariectomized mice as well as the involvement of aromatase, ERalpha, ERbeta, and AR in the effects. Ovariectomized and sham BALB/c mice were given daily treatment with either DHEA or E2 for three months, respectively. Bone mineral density was determined by DEXA after the last treatment. Mice were necropsied in 3 months after the treatment to analyze the ultrastructure of their femur osteoblasts (OBs) with a transmission electron microscope (TEM); DHEA, DHEA sulfate (DHEAS) and E2 levels were assayed by EIA; production in vitro of E2 in the uterus or tibia was assayed to evaluate the profile of P450arom activity; ERalpha and ERbeta mRNA levels in the uterus and tibia were determined by real-time PCR. The primary murine OBs were treated with DHEA and E2, respectively for 72 h. Real-time polymerase chain reaction (PCR) and western blot were carried out to evaluate aromatase, ERalpha, ERbeta and AR expression in OBs. Both DHEA and E2 significantly improved BMD and OB ultrastructure; E2 but not DHEA has significantly increased uterus wet weight, endometrium epithelial and gland thickness. Dehydroepiandrosterone not only increased serum, femoral DHEA, DHEAS and E2 concentration, but also increased uterine DHEA and DHEAS other than E2 concentration in site, while E2 only increased serum, uterine and femoral E2 concentration, but failed to alter the concentrations of DHEA and DHEAS. Moreover, DHEA significantly increased tibia P450arom enzyme activity, while E2 increased uterine and tibia aromatase activity. Furthermore, DHEA increased uterine ERbeta and ERalpha, and ERbeta transcription in the tibia, while E2 increased ERalpha transcription in the uterus and tibia. Dehydroepiandrosterone increased aromatase, ERalpha, ERbeta and AR expression in OBs, and increased significantly, but E2 apparently decreased the ratio of ERbeta/ERalpha. Although both DHEA and E2 augment BMD, the proliferative effects of E2 on the endometrium and uterus reflect the different modes of action on bone and the uterus, indicating that the preferable stimulatory effect of DHEA on bone appears to the more potential clinical values than estrogens in prophylaxis and therapeutics for PMO. But applicability of the findings from rodents in humans needs further study.

Fetal mitral valve movement recording by pulsed ultrasound

A technique for the recording of the fetal mitral valve is described and evidence to confirm the origin of the echoes is presented. Our results based on over 500 recordings, and their potential clinical values are discussed.

Normal Serum Proteins in Gastric Juice

There has been considerable interest in the high molecular weight constituents of gastric juice, with reference to both their biochemi­ cal characteristics and physiologic signifi­ cance. It has been shown that gastric juice from normal subjects and patients with gastric disease is a complex solution of high molecular weight constituents with diverse biochemical, 1. 2 chromatographic,3-5 electro­ phoretic, 6 • 7 polarographic,8 and biologic 9 characteristics. It has also been demon­ strated that there are significant qualitative and quantitative differences between these components in health and disease.1° It is likely that the heterogeneity of gastric juice is even greater than is indicated by the above analytic techniques. One un evaluated technic for gastric juice analysis is the specific immunologic identification of these high molecular weight constituents. This preliminary report is concerned with the application of the immunologic tech­ niques of gel diffusion precipitin analysis and immunoelectrophoresis to the demon­ stration of normal serum proteins present in certain pathologic gastric juice specimens, and their potential clinical values as indica­ tors of gastric disease.