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.

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.

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.

[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.

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.

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.

Clinical evaluation of the levels of 12 cytokines in serum/plasma under various storage conditions using evidence biochip arrays.

Cytokines are a group of peptides which form a sophisticated network to modulate multiple cellular events. Within such a network, and through complex feedback mechanisms, cytokine functions are largely interdependent and closely associated with a number of pathological processes. In the present study, the EVIDENCE 180 system was used to study the effects of storage temperature and repeated freeze/thaw cycles on the concentration of 12 cytokines in various sample types. Samples were collected from 9 healthy volunteers and stored by 3 methods: gel, glass and lithium heparin (LH) tubes. Immediately following collection, the concentration of each cytokine in the samples was measured. Cytokine concentrations of the 3 sample types that did not undergo repeated freeze/thaw cycles were compared with those subjected to 1­10 freeze/thaw cycles. In addition, the dynamic changes of 6 sample types which were stored at 4˚C for 6 h to 6 days was analyzed. In addition, the within­ and between­run precision of 12 cytokines on the biochip array was evaluated. Interleukin (IL)­8, vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) concentrations were lower in plasma compared with serum. Cytokine levels in serum and plasma were affected by several freeze/thaw cycles with IL­1ß, ­4 and ­10 increasing significantly following 1 freeze/thaw cycle and remaining at stable increased levels for the duration of the additional 9 cycles. In separated serum samples in gel and glass tubes stored at 4˚C for 6 days, no difference in concentration of the 12 cytokines was identified. In the other 4 sample types, IL­8, VEGF, tumor necrosis factor α and EGF levels were altered when stored at 4˚C. Results indicate that the EVIDENCE 180 system is stable and plasma was observed as the best sample type to determine concentration of the 12 cytokines using this biochip array. Repeated freeze/thaw cycles and storage at 4˚C was identified to affect the concentration of the 12 cytokines. The current study demonstrates that repeated freeze/thaw cycles of samples must be avoid. In addition, results indicate that plasma or serum must be separated immediately following centrifugation and sample concentration should be measured as soon as possible.

[The study on metabolic difference of human body affected by active stress and passive stress under special events].

OBJECTIVE: To study the metabolic difference of body influenced by active stress and passive stress under special events. METHODS: To detect serum multiple biochemistry index of 57 earthquake rescue medical team and 13 victims of a natural calamity in Wenchuan earthquake by using Hitachi 7600 automatic analyzer. RESULTS: Stress affected biochemistry index deeply. To compared with rescue medical team, the serum ADA, ALP and TG of victims increased obviously and TP, ALB, MAO, Cr, UA, K, Na, Cl, Ca, ApoA1 and HDL decreased obviously. CONCLUSION: Many biochemistry index have been changed under stress and it relate with stress extent. The human body function status was better in active stress than in passive stress.

[Change of IL-4 and IL-6 levels in blood plasma of patients transplanted with allogeneic hematopoietic stem cells determined by protein chip and its relationship with aGVHD].

This study was aimed to explore the relationship between changes of IL-4, IL-6 levels in blood plasma of patients receiving allo-HSCT and aGVHD through dynamical detection. The IL-4 and IL-6 levels in peripheral blood were detected by protein chip and were observed for 10 weeks. The results indicated that the IL-4 level increased rapidly in 1 week after transplantation in aGVHD group and was higher than that of non aGVHD group (p<0.05), while at 7th week after transplantation IL-4 level increased rapidly in non-GVHD group and was higher than that of aGVHD group (p<0.01). There was a significant difference of IL-6 level between these two groups before transplantation. IL-6 level reached peak at 1st week after transplantation in aGVHD group and was higher than that of non-aGVHD group (p<0.01), while IL-6 level was significantly higher in non-aGVHD group than that of aGVHD group at 4th week after transplantation (p<0.01) and at 5th week after transplantation (p<0.05). It is concluded that the levels of IL-4 and IL-6 had been increased rapidly after hematopoietic stem cell transplantation, indicating that aGVHD will occur. The high level of IL-6 before transplantation may be the risk factor of aGVHD occurrence after transplantation. aGVHD may occur later if the blood plasma IL-4 level rises in patients without aGVHD. Therefore, dynamically monitoring IL-4 and IL-6 levels contributes to predict the occurrence of aGVHD.