Both epilepsy and anti-seizure medications affect bone metabolism in children with self-limited epilepsy with centrotemporal spikes.

OBJECTIVE: Bone metabolism can be influenced by a range of factors. We selected children with self-limited epilepsy with centrotemporal spikes (SeLECTS) and lifestyles similar to those of healthy children to control for the confounding factors that may influence bone metabolism. We aimed to identify the specific effects of epilepsy and/or anti-seizure medications (ASMs) on bone metabolism. METHODS: Patients with SeLECTS were divided into an untreated group and a monotherapy group, and the third group was a healthy control group. We determined the levels of various biochemical markers of bone metabolism, including procollagen type I nitrogenous propeptide (PINP), alkaline phosphatase (ALP), osteocalcin (OC), collagen type I cross-linked C-telopeptide (CTX), calcium, magnesium, phosphorus, parathyroid hormone (PTH), and vitamin D3 (VD3 ). RESULTS: A total of 1487 patients (from 19 centers) were diagnosed with SeLECTS; 1032 were analyzed, including 117 patients who did not receive any ASMs (untreated group), 643 patients who received only one ASM (monotherapy group), and 272 children in the healthy control group. Except for VD3 , other bone metabolism of the three groups were different (p < .001). Bone metabolism was significantly lower in the untreated group than the healthy control group (p < .05). There were significant differences between the monotherapy and healthy control group in the level of many markers. However, when comparing the monotherapy and untreated groups, the results were different; oxcarbazepine, levetiracetam, and topiramate had no significant effect on bone metabolism. Phosphorus and magnesium were significantly lower in the valproic acid group than the untreated group (adjusted p < .05, Cliff's delta .282-.768). CTX was significantly higher in the lamotrigine group than in the untreated group (adjusted p = .012, Cliff's delta = .316). SIGNIFICANCE: Epilepsy can affect many aspects of bone metabolism. After controlling epilepsy and other confounders that affect bone metabolism, we found that the effects of ASMs on bone metabolism differed. Oxcarbazepine, levetiracetam, and topiramate did not affect bone metabolism, and lamotrigine corrected some of the abnormal markers of bone metabolism in patients with epilepsy.

Blood index panel for gastric cancer detection.

BACKGROUND: Gastric cancer is a common malignant tumor. Early detection and diagnosis are crucial for the prevention and treatment of gastric cancer. AIM: To develop a blood index panel that may improve the diagnostic value for discriminating gastric cancer and gastric polyps. METHODS: Thirteen tumor-related detection indices, 38 clinical biochemical indices and 10 cytokine indices were examined in 139 gastric cancer patients and 40 gastric polyp patients to build the model. An additional 68 gastric cancer patients and 22 gastric polyp patients were enrolled for validation. After area under the curve evaluation and univariate and multivariate analyses. RESULTS: Five tumor-related detection indices, 12 clinical biochemical indices and 1 cytokine index showed significant differences between the gastric cancer and gastric polyp groups. Carbohydrate antigen (CA) 724, phosphorus (P) and ischemia-modified albumin (IMA) were included in the blood index panel, and the area under the curve (AUC) of the index panel was 0.829 (0.754, 0.905). After validation, the AUC was 0.811 (0.700, 0.923). Compared to the conventional index CA724, the blood index panel showed significantly increased diagnostic value. CONCLUSION: We developed an index model that included CA724, P and IMA to discriminate the gastric cancer and gastric polyp groups, which may be a potential diagnostic method for clinical practice.

Droplet digital polymerase chain reaction assay for methylated ring finger protein 180 in gastric cancer.

BACKGROUND: Gastric cancer (GC) is one of the most prevalent malignant tumors that endangers human health. Early diagnosis is essential for improving the prognosis and survival rate of GC patients. Ring finger protein 180 (RNF180) is involved in the regulation of cell differentiation, proliferation, apoptosis, and tumorigenesis, and aberrant hypermethylation of CpG islands in the promoter is strongly associated with the occurrence and development of GC. Thus, methylated RNF180 can be used as a potential biomarker for GC diagnosis. AIM: To use droplet digital polymerase chain reaction (ddPCR) to quantify the methylation level of the RN180 gene. A reproducible ddPCR assay to detect methylated RNF180 from trace DNA was designed and optimized. METHODS: The primer and probe were designed and selected, the conversion time of bisulfite was optimized, the ddPCR system was adjusted by primer concentration, amplification temperature and amplification cycles, and the detection limit of ddPCR was determined. RESULTS: The best conversion time for blood DNA was 2 h 10 min, and that for plasma DNA was 2 h 10 min and 2 h 30 min. The results of ddPCR were better when the amplification temperature was 56 °C and the number of amplification cycles was 50. Primer concentrations showed little effect on the assay outcome. Therefore, the primer concentration could be adjusted according to the reaction system and DNA input. The assay required at least 0.1 ng of input DNA. CONCLUSION: In summary, a ddPCR assay was established to detect methylated RNF180, which is expected to be a new diagnostic biomarker for GC.

Effect of Leptin Levels of Type 2 Diabetes Mellitus and Diabetic Complications.

PURPOSE: To investigate serum leptin levels in patients with type 2 diabetes mellitus (T2DM) and the relationship between leptin levels and T2DM complications and prevalence. METHODS: A total of 355 patients, 282 cases with T2DM and 73 normal controls, were recruited at 1st Medical Centre, Chinese PLA General Hospital (Beijing, China) between November 2013 and July 2014. Levels of serum leptin, biochemical markers and sexual hormones were measured, and clinical characteristics were retrieved through the electronic medical record system. RESULTS: Leptin levels in females were higher than that in males. Leptin levels in T2MD patients were positively correlated with body mass index, percent body fat, triglyceride, cystatin C homocysteine and salivary acid, and negatively correlated with glycosylated serum protein and glycosylated albumin levels. Leptin levels in males were positively correlated with systolic pressure and estradiol, and negatively correlated with testosterone and high density lipoprotein cholesterol. Sex (female) was positively correlated with the duration of disease. Leptin levels in T2DM patients with complications such as hypertension, diabetic nephropathy, diabetic peripheral neuropathy and coronary heart disease were higher than that in patients without such complications. Leptin levels in females with diabetic retinopathy and diabetic macroangiopathy were higher than that in patients without such complications, but there was no difference in males. CONCLUSIONS: Leptin has significant gender differences. Leptin levels are related to body mass index, percent body fat and sex hormone level in T2DM patients and may affect short-term blood glucose control in T2DM patients. Leptin levels are related to complications in patients with T2DM and affect the prevalence rates of complications.

Reference Intervals and Regularity Analysis of Biochemical Makers in 3-Year-Old Children.

BACKGROUND: The reference intervals of biochemical markers were significantly affected by age and gender, especially in minors. In recent years, many provinces and regions in China had established reference intervals for children's hematological indicators. Without considering the instruments and reagents, the reference interval will also be affected by the region, economic development, eating habits, and other factors. Therefore, the reference interval of any hematological indicators is not necessarily a fixed range, it changes with certain factors. In our study, we analyzed the changes of biochemical markers in different serum total Ca and 25-OH-D concentrations, and established the reference intervals of biochemical markers in 3-year-old children, explored the change trend of biochemical markers with different serum total Ca and 25-OH-D concentration. METHODS: Data was collected from 226 cases of 3-year-old children for biochemical markers, in the Chinese PLA General Hospital in August 2015. The data were divided into a high-level group (serum total Ca > 2.63 mmol/L and 25-OH-D > 40.81 ng/mL) and a low-level group (serum total Ca < 2.54 mmol/L and 25-OH-D < 32.64 ng/mL) according to serum total Ca and 25-OH-D levels for comparison. The change trend of biochemical markers was compared according to serum total Ca and 25-OH-D level. RESULTS: The Glu levels in boys were significantly higher than that in girls, but CHO and LDL-C in girls were significantly higher than that in boys. The reference intervals of ATL (5.6 - 22.1 U/L), ALB (44.8 - 55.2 g/L), TP (62.7 - 83.1 g/L), ALP (154.4 - 379.7 U/L), GGT (7.2 - 15.9 U/L), Glu (boys: 4.08 - 5.91 mmol/L; girls: 4.05 - 5.37 mmol/L), UREA (2.7 - 6.3 mmol/L), CREA (26.4 - 46.8 µmol/L), UA (182.4 - 400.2 µmol/L), TG (0.43 - 1.67 mmol/L), CHO (boys: 3.19 - 5.96 mmol/L; girls: 3.03 - 6.51 mmol/L), HDL-C (0.98 - 2.24 mmol/L), LDL-C (boys: 1.30 - 3.64 mmol/L; girls: 1.24 - 4.27 mmol/L), total Ca (2.34 - 2.85 mmol/L), PHOS (1.38 - 2.06 mmol/L), Mg (0.83 - 1.06 mmol/L), osteocalcin (41.64 - 91.92 ng/mL), PTH (12.08 - 43.06 pg/mL), 25-OH-D (19.66 - 56.37 ng/mL), ß-CrossLaps (0.82 - 1.88 ng/mL), TP1NP (357.9 - 1025.7 µg/L) were established. ALT, TP, ALB, GGT, Glu, CHO, HDL-C, LDL-C, UREA, CREA, PHOS, Mg, and ALP in high level group were significantly higher than those in low level group. There was no significant difference in TG, UA, TP1NP, osteocalcin, PTH and ß-CrossLaps between high level group and low-level group. With the increase of serum total Ca and 25-OH-D levels, most of the biochemical markers had a gradually increasing trend. However, biochemical markers of bone (TP1NP, osteocalcin, PTH, ß-CrossLaps) showed different trends. CONCLUSIONS: This study established the reference intervals of biochemical markers of 3-year-old children. The changes of serum total Ca and 25-OH-D levels in children reflected the changes of glucose and lipid metabolism, liver and kidney function markers, and indirectly reflected the growth and development of children and various organ functions. Maintaining high levels of serum total Ca and 25-OH-D can promote the growth and development of children.

Development and Evaluation of a Combined Immunoassay Method Based on a Stable Isotope Tagging Strategy and Inductively Coupled Plasma Mass Spectrometry for Detecting Human Chorionic Gonadotropin.

BACKGROUND: The development of a combined immunoassay method, based on a stable isotope tagging strategy and inductively coupled plasma mass spectrometry (ICP-MS), has created options for quantitative bioanalysis. The aim of the study was to develop a combined immunoassay, featuring ICP-MS and a stable element labeling strategy, for the detection of human chorionic gonadotropin (HCG), and developed methodology applicable for clinical practice. METHODS: In accordance with guidelines published by the Clinical and Laboratory Standards Institute (CLSI), we developed our assay and then evaluated its analytical performance, including the limit of detection (LOD), the upper limit of quantification (ULoQ), linearity, precision, recovery, cross reactivity, and interference. Next, we collected 130 clinical samples for analysis with the new assay. The data derived from our assay were then compared with those derived by an existing electrochemiluminescence immunoassay (ECLIA). RESULTS: The LOD of the assay was 0.33 mIU/mL and the ULoQ was 11,300 mIU/mL. The coefficient of determina-tion of linearity was higher than 0.99 in the range of 1 to 8,917 mIU/mL (R2 = 0.9964). The obtained recoveries ranged from 97.08% to 103.50%, while the intra-assay imprecision of high value samples and low value samples were 2.97% and 6.08%, respectively. The inter-assay imprecision of high value samples and low value samples were 3.98% and 7.08%, respectively. Interference test results deviated by less than ± 10% in the presence of hemoglobin ≤ 2 g/L, bilirubin ≤ 274 mol/L, or triglycerides ≤ 37 mmol/L. Compared with the commercial ECLIA method for clinical sample detection, the proposed method showed a significant correlation (R2 = 0.9770) and satisfactory agreement. CONCLUSIONS: The combination of ICP-MS and a stable element labeling based immunoassay for HCG detection was established successfully and the general performance of this system was acceptable, thus indicating that the assay has potential for the clinical application.

Serum exosomal microRNAs combined with alpha-fetoprotein as diagnostic markers of hepatocellular carcinoma.

Exosomal microRNAs have recently been studied as the potential diagnostic marker for various malignancies, including hepatocellular carcinoma (HCC). The aim of this study was to investigate serum exosomal microRNA profiles as HCC diagnostic marker. Transmission electron microscopy and Western blot were used to identify serum exosomes. Deep sequencing was performed to screen differentially expressed microRNAs between HCC (n = 5) and liver cirrhosis (LC, n = 5) groups. Three upregulated and two downregulated microRNAs were selected for qPCR analysis. The levels of selected microRNAs were normalized to Caenorhabditis elegans miR-39 microRNA mimics. Serum exosomal level of miR-122, miR-148a, and miR-1246 was further analyzed and significantly higher in HCC than LC and normal control (NC) groups (P < 0.001), but not different from chronic hepatitis group (P > 0.05). The receiver operating characteristic curve was used to evaluate the diagnostic performance of candidate microRNAs. Area under the curve (AUC) of miR-148a was 0.891 [95% confidence interval (CI), 0.809-0.947] in discriminating HCC from LC, remarkably higher than alpha-fetoprotein (AFP) (AUC: 0.712, 95% CI: 0.607-0.803). Binary logistic regression was adopted to establish the diagnostic model for discriminating HCC from LC. And the combination of miR-122, miR-148a, and AFP increased the AUC to 0.931 (95% CI, 0.857-0.973), which can also be applied for distinguishing early HCC from LC. miR-122 was the best for differentiating HCC from NC (AUC: 0.990, 95% CI, 0.945-1.000). These data suggest that serum exosomal microRNAs signature or their combination with traditional biomarker may be used as a suitable peripheral screening tool for HCC.

Relative Handgrip Strength Is Inversely Associated with Metabolic Profile and Metabolic Disease in the General Population in China.

Background: Absolute handgrip strength has been correlated with metabolic profile and metabolic disease. Whether relative handgrip strength is also associated with metabolic disease has not been assessed. This study aimed at assessing the association of relative handgrip strength with metabolic profile and metabolic disease in the general population in China. Methods: Data were derived from an ongoing cross-sectional survey of the 2013 National Physical and Health in Shanxi Province, which involved 5520 participants. Multiple linear regression or multiple logistic regression analysis were used to assess the association of absolute/relative handgrip strength with the metabolic profile, preclinical, and established stages of metabolic diseases. Results: This study revealed that relative handgrip strength, that is when normalized to BMI, was associated with: (1) in both genders for more favorable blood lipid levels of high-density lipoprotein cholesterol [males: b = 0.19 (0.15, 0.23); females: b = 0.22 (0.17, 0.28)], low-density lipoprotein cholesterol [males: b = -0.14 (-0.23, -0.05); females: b = -0.19 (-0.31, -0.18)], triglycerides [males: b = -0.58 (-0.74, -0.43); females: b = -0.55 (-0.74, -0.36)] and total cholesterol [males: b = -0.20 (-0.31, -0.10); females: b = -0.19 (-0.32, -0.06)]; and better serum glucose levels in males [b = -0.30 (-0.46, -0.15)]. (2) lower risk of impaired fasting glucose in males {third quartile [OR = 0.66 (0.45-0.95)] and fourth quartile [OR = 0.46 (0.30-0.71)] vs. first quartile} and dyslipidemia in both genders {third quartile [males: OR = 0.65 (0.48-0.87); females: OR = 0.68 (0.53-0.86)] and fourth quartile [males: OR = 0.47 (0.35-0.64); females: OR = 0.47(0.36-0.61)] vs. first quartile}. (3) lower risk of hyperlipidemia in both genders third quartile [males: OR = 0.66 (0.50-0.87); females: OR = 0.57 (0.43-0.75)] and fourth quartile [males: OR = 0.35 (0.26-0.47); females: OR = 0.51 (0.38-0.70)] vs. first quartile. However, contrary to relative handgrip strength, higher absolute handgrip strength was associated with unfavorable metabolic profiles and higher risk of metabolic diseases. These paradoxical associations were retained even after adjusted for BMI by employed a multivariate analysis. Conclusion: We conclude that measurement of relative handgrip strength can be used as a reasonable clinical predictor of metabolic health and disease.

Dynamic changes in serum cytokine levels and their clinical significance in predicting acute GVHD.

To explore the clinical significance of cytokines and biochemical tests in acute graft-versus-host disease (aGVHD), we detected the concentrations of 8 cytokines and 19 conventional biochemical markers in the sera of aGVHD and non-GVHD patients throughout the process of allogeneic hematopoietic stem cell transplantation and the onset of aGVHD. Predictive models were then established using the 27 indices, and models were verified by a prospective trial. The 27 indices showed significant differences between aGVHD patients and non-GVHD control subjects (two-tailed p<0.05) prior to transplantation and before the onset of aGVHD. Our models, established by binary logistic regression on days +7 and +14, showed a significant absolute capacity of predicting grade 2∼4 aGVHD with positive and negative predictive values of at least 70%. Our data showed that the progression of aGVHD could induce dynamic changes in the levels of serum cytokines and biochemical markers. Because most of these tests were less specific for aGVHD, these changes were easily neglected in clinical work. However, by combining cytokine and biochemical tests, the established prediction model can greatly improve the ability of these biomarkers to predict the development of aGVHD one or two weeks earlier.

Association of prehypertension and cardiovascular risk factor clustering in Inner Mongolia: a cross-sectional study.

OBJECTIVES: To assess the clustering of cardiovascular disease (CVD) risk factors in Han and Mongolian adults with prehypertension or hypertension in Northern China. METHODS: We selected 3227 Han and Mongolian participants (20-80 years old) using a multistage cluster sampling method in 2014. The participants were interviewed by standard questionnaires and underwent anthropometric measurement and biochemical testing. Han and Mongolian participants were divided into optimal, prehypertension, and hypertension groups based on blood pressure. A multinomial logit analysis was performed to explore relationships between CVD risk factor clustering and prehypertension or hypertension, and the heterogeneity between Han and Mongolian was evaluated by the Cochran Q test. The differences between the ethnic groups in the proportions of risk factors was tested with the χ2 test. RESULTS: The clustering of two or three CVD risk factors in the prehypertension or hypertension groups was consistently higher than in the optimal group (Bonferroni, p<0.0167). The odds ratios (ORs) of prehypertension and hypertension increased with the number of CVD risk factors (ptrend <0.0001). In multivariate modelling, the adjusted ORs of one, two, and ≥3 CVD risk factors versus no risk factors was, respectively, 1.95, 2.25, and 2.28 in Han prehypertensive participants, and 1.73, 2.83, and 3.69 in Mongolian prehypertensive participants. In addition, the adjusted ORs were 3.15, 4.75, and 6.49 in Han hypertensive participants, and 1.90, 5.29, and 8.13 in Mongolian hypertensive participants (all p<0.05). There was no significant heterogeneity between Han and Mongolian participants in the prehypertension or hypertension groups. The age-standardised prevalence of ≥3 risk factors was 38.30% in Han men and 39.79% in Mongolian men. The rate was significantly lower in Han women than Mongolian women (9.18% vs 14.55%, p=0.002). CONCLUSIONS: These findings showed clustering of CVD risk factors in prehypertensive Han and Mongolian adults, and showed prehypertension may be a useful target for intervention.

Preoperative Serum Carcinoembryonic Antigen as a Marker for Predicting the Outcome of Three Cancers.

BACKGROUND: Serum levels of carcinoembryonic antigen (CEA) are associated with a variety of tumors. OBJECTIVE: This study evaluated the prognostic value of pretreatment serum CEA levels in predicting the outcomes of multiple tumors subjected to treatment. METHODS: Prior to therapy, serum samples from 71 prostate, 46 breast, 77 gastric, and 31 pancreatic cancer patients were collected to examine serum CEA levels. The cutoff value for CEA was set as determined by the maximum Youden index. The data were analyzed by the Kaplan-Meier curves generated by the log-rank test and Cox multivariate analysis. RESULTS: The overall survival rate for all the patients was 71.11%. The 3-year survival rate of patients with prostate, breast, gastric, and pancreatic cancers was 81.69%, 95.65%, 54.55%, and 51.61%, respectively. The 3-year survival rate showed significant statistical differences between patients with serum CEA levels <2.885 µg/L and those with serum CEA levels ⩾2.885 µg/L (P < .001). The statistical differences of the 3-year survival rate also existed in the men (P = .010) or women group (P < .001), as well as in the 3 different types of cancer, which include breast cancer (P = .025), gastric cancer (P = .001), and pancreatic cancer (P = .047). CONCLUSIONS: Serum CEA levels can provide additional prognostic information and may be useful in treatment implementation for patients with breast, gastric, or pancreatic cancer.

[Determination of 20 Trace Elements in the Blood Collection Tubes with ICP-MS].

To investigate the contamination of blood collection tubes, 20 trace elements (Al, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Mo, Cd, Sn, Sb, Ba, W, Hg, Tl, Pb) in 13 different types of blood collection tube were studied with ICP-MS method. The lixivium of H(2)O and 10% HNO(3) were measured with ICP-MS, and then the contamination coming from the blood collection tube is specified. According to the concentration range of human blood, plasma and serum from recently published literature, this report presents a detailed analysis of capable trace elements for each blood collection tube. The results showed that, tube No.1 is capable to analyze 18 trace elements in the human serum; tube No.6 is capable to analyze 15 trace elements in the human plasma; tube No. 13 is capable to analyze 17 trace elements in the human blood. But we still should be aware that, the elements Sb and W in tube No.1, the elements V, Cr, Ni, and Sb in tube No.6, and the elements Al, Sb and W in tube No.13, are in the same magnitude of the normal trace element concentration range in the human serum, plasma and blood. They might affect the testing results. The serum collected from the same volunteer by tube No.1 and tube No.3 were compared here, the results show that, almost each trace element concentration of human serum from tube No.1 is lower than from tube No.3, especially for elements Al, V, Cr, Mn, As, Sn, and Sb. The results indicate that the blood collection tubes show great impact on determination of trace element.

Association between clusterin polymorphisms and esophageal squamous cell carcinoma risk in Han Chinese population.

Genetic susceptibility plays an essential role in an individual's risk of esophageal squamous cell carcinoma (ESCC). The aim of this study is to investigate the associations between clusterin (CLU) gene polymorphisms and ESCC risk. We undertook a case-control study to analyze three CLU polymorphisms (gene rs9331888 C>G, rs17466684 A>G and rs1532278 T>C) in an Han Chinese population, by extraction of genomic DNA from the peripheral blood of 642 patients with ESCC and 658 control participants, and performed CLU genotyping using DNA sequencing. The obtained results indicated that overall, no statistically significant association was observed in rs17466684 and rs1532278. However, gene rs9331888 C>G genotype was at increased risk of ESCCs (P=0.037; odds ratio (OR)=1.089, 95% CI: 1.006-1.175). Moreover, rs9331888 G/G genotype ESCCs were more significantly common in patients with tumor size of >5 cm than T allele ESCC and in cases of poor differentiation and lower advanced pathological stage. In conclusion, polymorphism in rs9331888 C>G was observed to be associated with susceptibility of ESCC. Nevertheless, further investigation with a larger sample size is needed to support our results.

A randomized controlled clinical trial of formalin for treatment of chronic hemorrhagic radiation proctopathy in cervical carcinoma patients.

PURPOSE: Chronic hemorrhagic radiation proctopathy is not uncommon after radiotherapy for cervical carcinoma. The outcomes of several treatments have been variable. Many studies demonstrate that topical treatment with 4 % formalin is effective and safe. However, a nonrandomized control study showed a high response rate and good tolerance in chronic radiation proctopathy patients treated with 10 % formalin. The optimal concentration of formalin therefore remains unclear. METHODS: To compare the effectiveness and safety of 4 and 10 % formalin for the treatment of chronic hemorrhagic radiation proctopathy, a prospective trial was conducted at the Department of Gynecology of the Affiliated Hospital of Binzhou Medical College from January 2009 to December 2012. One hundred and twenty patients with chronic hemorrhagic radiation proctopathy following radiotherapy for cervical carcinoma were recruited and randomized to receive 4 or 10 % formalin. A standard protocol was followed for formalin application. Symptom and rectoscopy scores were evaluated before and at 12 weeks after treatment. RESULTS: In the 4 % formalin group, 49 (86.0 %) and 53 (91.4 %) patients showed an improvement in symptom score and rectoscopy score, respectively (P = 0.36). Symptom and rectoscopy scores decreased significantly after treatment in both the 4 % formalin group and the 10 % formalin group (P < 0.001). Symptom score was correlated with rectoscopy score (P < 0.001). More patients in the 10 % group suffered treatment-related complications than did those in the 4 % group (P = 0.03). CONCLUSIONS: For the treatment of chronic hemorrhagic radiation proctopathy, 4 % should be the preferred formalin concentration.

Identification and characterization of novel serum microRNA candidates from deep sequencing in cervical cancer patients.

Small non-coding microRNAs (miRNAs) are involved in cancer development and progression, and serum profiles of cervical cancer patients may be useful for identifying novel miRNAs. We performed deep sequencing on serum pools of cervical cancer patients and healthy controls with 3 replicates and constructed a small RNA library. We used MIREAP to predict novel miRNAs and identified 2 putative novel miRNAs between serum pools of cervical cancer patients and healthy controls after filtering out pseudo-pre-miRNAs using Triplet-SVM analysis. The 2 putative novel miRNAs were validated by real time PCR and were significantly decreased in cervical cancer patients compared with healthy controls. One novel miRNA had an area under curve (AUC) of 0.921 (95% CI: 0.883, 0.959) with a sensitivity of 85.7% and a specificity of 88.2% when discriminating between cervical cancer patients and healthy controls. Our results suggest that characterizing serum profiles of cervical cancers by Solexa sequencing may be a good method for identifying novel miRNAs and that the validated novel miRNAs described here may be cervical cancer-associated biomarkers.

Role of EZH2 polymorphisms in esophageal squamous cell carcinoma risk in Han Chinese population.

Gene single nucleotide polymorphisms play a critical role in the development of esophageal squamous cell carcinoma (ESCC). The aim of this study is to investigate the associations between EZH2 gene polymorphisms and ESCC risk. We undertook a case-control study to analyze three EZH2 polymorphisms (148505302C>T, 2110+6A>C and 626-394T>C) in an Han Chinese population, by extraction of genomic DNA from the peripheral blood of 476 patients with ESCC and 492 control participants, and performed EZH2 genotyping using DNA sequencing. The obtained results indicated that overall, no statistically significant association was observed in 148505302C>T and 2110+6A>C. However, 626-394T>C genotype was at increased risk of ESCCs (p=0.006; odds ratio (OR)=1.131, CI 95%: 1.034-1.236). Moreover, 626-394C/C genotype ESCCs were more significantly common in patients with tumor size of >5 cm than T allele ESCC and in cases of poor differentiation and lower advanced pathological stage. In conclusion, polymorphism in 626-394T>C was observed to be associated with susceptibility of ESCC. Nevertheless, further investigation with a larger sample size is needed to support our results.

TESTIN suppresses tumor growth and invasion via manipulating cell cycle progression in endometrial carcinoma.

BACKGROUND: The TESTIN gene was demonstrated to be a tumor suppressor in prostate and breast cancer through inhibiting tumor growth and invasion. Herein, we aimed to investigate the detailed functions of TESTIN in the highly sexual hormone (estrogen)-dependent malignancy, endometrial carcinoma. MATERIAL AND METHODS: TESTIN mRNA and protein expression were measured by qRT-PCR, Western blot and immunohistochemistry. Upregulation of TESTIN was achieved by transfecting the pcDNA3.1-TESTIN plasmids into AN3CA cells. Knockdown of TESTIN was achieved by transfecting the shRNA-TESTIN into Ishikawa cells. MTT assay, colony formation assay, and Transwell assay were used to investigate the effects of TESTIN on cellular proliferation and invasion. The apoptotic status and cell cycle were analyzed using flow cytometry. MMP2 secretion was determined by ELISA assay. The xenograft assay was used to investigate the functions of TESTIN in nude mice. RESULTS: Compared to the non-malignant adjacent endometrium, 54% of tumor samples presented downregulation of TESTIN (P<0.001). Loss of TESTIN protein was correlated with advanced tumor stage (P=0.047), high grade (P=0.034), and lymphatic vascular space invasion (P=0.036). In vitro, overexpression of TESTIN suppressed cell proliferation, induced dramatic G1 arrest, and inhibited tumor invasion through blocking the secretion of MMP2. Loss of TESTIN accelerated cellular proliferation, promoted cell cycle progression, and enhanced tumor invasion by increasing the secretion of MMP2. Consistently, TESTIN could significantly delay the growth of xenografts in nude mice. CONCLUSIONS: TESTIN was commonly downregulated in human endometrial carcinoma and was associated with poor prognostic markers. Moreover, TESTIN significantly inhibited tumor growth and invasion via arresting cell cycle in in vitro and in vivo experiments. Therefore, we propose that TESTIN might be a prognostic marker and therapeutic target for endometrial carcinoma.

Association between EZH2 polymorphisms and colorectal cancer risk in Han Chinese population.

The aim of this study is to investigate the associations between EZH2 gene polymorphisms and colorectal cancer (CRC) risk. We undertook a case-control study to analyze three EZH2 polymorphisms (148505302C>T, 2110+6A>C and 626-394T>C) in an Han Chinese population, by extraction of genomic DNA from the peripheral blood of 512 patients with CRC and 546 control participants, and performed EZH2 genotyping using DNA sequencing. The obtained results indicated that overall, no statistically significant association was observed in 2,110+6A>C. Nevertheless, 148505302C>T genotype demonstrated a protective effect in CRCs (P=0.014; odds ratio (OR) 0.777, CI 95%:0.647-0.933). Furthermore, 148505302 T allele CRC was more significantly common in patients with tumor size of <4 cm than C allele CRC and in cases of good differentiation and lower advanced pathological stage. However, 626-394T>C genotype was at increased risk of CRCs (P<0.001; odds ratio (OR) 1.457, CI 95%:1.160-1.829). Moreover, 626-394C/C genotype CRCs were more significantly common in patients with tumor size of >4 cm than T allele CRC and in cases of poor differentiation and lower advanced pathological stage. In conclusion, polymorphism in 626-394T>C was observed to be associated with susceptibility of CRC. However, 148505302C>T polymorphism indicated to play a protective role in susceptibility to CRC. Nevertheless, further investigation with a larger sample size is needed to support our results.

A novel differential predict model based on matrix-assisted laser ionization time-of-flight mass spectrometry and serum ferritin for acute graft-versus-host disease.

Clinical diagnosis of acute graft-versus-host disease (aGVHD) mainly depends on clinical manifestation and tissue biopsies, leading to a delayed diagnosis and treatment for aGVHD patients when the early symptom is insignificant. Our objective was to investigate the possibility of prewarning the risk of aGVHD before and after allogeneic hematopoietic stem cell transplantation (allo-HSCT) by serum protein profiling combined with serum ferritin. The difference in polypeptide expression before and after transplantation had been compared by using CLINPROT technology, and serum ferritin levels have been analyzed simultaneously. Through combining serum ferritin and MS spectral data, the diagnosis sensitivity and specificity of our model for prewarning severe aGVHD (III~IV°aGVHD) before transplant all increased to 90.0%, while after transplant, the sensitivity and specificity are 78.3% and 86.4%. Our joint prewarning model could predict the risk of aGVHD, especially severe aGVHD before and after transplant, which also provides a reliable method to the continuous monitoring of the condition of patients.

Role of KAI1/CD82 polymorphisms in colon cancer risk in Han Chinese population.

The aim of this study is to investigate the associations between KAI1/CD82 gene polymorphisms and colorectal cancer (CRC)-risk predisposition. We undertook a case-control study to analyze two KAI1/CD82 polymorphisms (exon 3 -29166 C>T and exon 9 -52840 C>A) in an Han Chinese population, by extraction of genomic DNA from the peripheral blood of 356 patients with CRC and 378 control participants, and performed KAI1/CD82 genotyping using DNA sequencing. The obtained results indicated that overall, no statistically significant association was observed in exon 9 (-52840 C>A). Nevertheless, exon 3 (-29166 C>T) genotype was at increased risk of CRC (P = 0.006; odds ratio = 1.299, CI 95% 1.058-1.549). Furthermore, -29166 T allele CRCs were more significantly common in patients with tumor size of >4 cm than C allele CRC and in cases of poor differentiation and advanced pathological stage. These findings led us to conclude that polymorphism in exon 3 (-29166 C>T) was observed to be associated with susceptibility of CRC. However, exon 9 (-52840 C>A) polymorphism showed no correlation to CRC susceptibility. Nevertheless, further investigation with a larger sample size is needed to support our results.