Clinical evaluation of a multitarget fecal immunochemical test-sDNA test for colorectal cancer screening in a high-risk population: a prospective, multicenter clinical study.

Colorectal cancer (CRC) is a major malignancy threatening the health of people in China and screening could be effective for preventing the occurrence and reducing the mortality of CRC. We conducted a multicenter, prospective clinical study which recruited 4,245 high-risk CRC individuals defined as having positive risk-adapted scores or fecal immunochemical test (FIT) results, to evaluate the clinical performance of the multitarget fecal immunochemical and stool DNA (FIT-sDNA) test for CRC screening. Each participant was asked to provide a stool sample prior to bowel preparation, and FIT-sDNA test and FIT were performed independently of colonoscopy. We found that 186 (4.4%) were confirmed to have CRC, and 375 (8.8%) had advanced precancerous neoplasia among the high CRC risk individuals. The sensitivity of detecting CRC for FIT-sDNA test was 91.9% (95% CI, 86.8-95.3), compared with 62.4% (95% CI, 54.9-69.3) for FIT (P < 0.001). The sensitivity for detecting advanced precancerous neoplasia was 63.5% (95% CI, 58.3-68.3) for FIT-sDNA test, compared with 30.9% (95% CI, 26.3-35.6) for FIT (P < 0.001). Multitarget FIT-sDNA test detected more colorectal advanced neoplasia than FIT. Overall, these findings indicated that in areas with limited colonoscopy resources, FIT-sDNA test could be a promising further risk triaging modality to select patients for colonoscopy in CRC screening.

Elevated serum anti-Saccharomyces cerevisiae antibody accompanied by gut mycobiota dysbiosis as a biomarker of diagnosis in patients with de novo Parkinson disease.

BACKGROUND AND PURPOSE: Intestinal inflammation and gut microbiota dysbiosis contribute to Parkinson disease (PD) pathogenesis, and growing evidence suggests associations between inflammatory bowel diseases (IBD) and PD. Considered as markers of chronic gastrointestinal inflammation, elevated serum anti-Saccharomyces cerevisiae antibody (ASCA) levels, against certain gut fungal components, are related to IBD, but their effect on PD is yet to be investigated. METHODS: Serum ASCA IgG and IgA levels were measured using an enzyme-linked immunosorbent assay, and the gut mycobiota communities were investigated using ITS2 sequencing and analyzed using the Qiime pipeline. RESULTS: The study included 393 subjects (148 healthy controls [HCs], 140 with PD, and 105 with essential tremor [ET]). Both serum ASCA IgG and IgA levels were significantly higher in the PD group than in the ET and HC groups. Combining serum ASCA levels and the occurrence of constipation could discriminate patients with PD from controls (area under the curve [AUC] = 0.81, 95% confidence interval [CI] = 0.76-0.86) and from patients with ET (AUC = 0.85, 95% CI = 0.79-0.89). Furthermore, the composition of the gut fungal community differed between the PD and HC groups. The relative abundances of Saccharomyces cerevisiae, Aspergillus, Candida solani, Aspergillus flavus, ASV601_Fungi, ASV866_Fungi, and ASV755_Fungi were significantly higher in the PD group, and enriched Malassezia restricta was found in the HC group. CONCLUSIONS: Our study identified elevated serum ASCA levels and enriched gut Saccharomyces cerevisiae in de novo PD.

Different patterns of exosomal α-synuclein between Parkinson's disease and probable rapid eye movement sleep behavior disorder.

BACKGROUND AND PURPOSE: The insidious onset of Parkinson's disease (PD) makes early diagnosis difficult. Notably, idiopathic rapid eye movement sleep behavior disorder (iRBD) was reported as a prodrome of PD, which may represent a breakthrough for the early diagnosis of PD. However, currently there is no reliable biomarker for PD diagnosis. Considering that α-synuclein (α-Syn) and neuroinflammation are known to develop prior to the onset of clinical symptoms in PD, it was hypothesized that plasma total exosomal α-Syn (t-exo α-Syn), neural-derived exosomal α-Syn (n-exo α-Syn) and exosomal apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC) may be potential biomarkers of PD. METHODS: In this study, 78 PD patients, 153 probable iRBD patients (pRBD) and 63 healthy controls (HCs) were recruited. α-Syn concentrations were measured using a one-step paramagnetic particle-based chemiluminescence immunoassay, and ASC levels were measured using the Ella system. RESULTS: It was found that t-exo α-Syn was significantly increased in the PD group compared to the pRBD and HC groups (p < 0.0001), whilst n-exo α-Syn levels were significantly increased in both the PD and pRBD groups compared to HCs (p < 0.0001). Furthermore, although no difference was found in ASC levels between the PD and pRBD groups, there was a positive correlation between ASC and α-Syn in exosomes. CONCLUSIONS: Our results suggest that both t-exo α-Syn and n-exo α-Syn were elevated in the PD group, whilst only n-exo α-Syn was elevated in the pRBD group. Additionally, the adaptor protein of inflammasome ASC is correlated with α-Syn and may facilitate synucleinopathy.

Integrated Analysis to Obtain Potential Prognostic Signature in Glioblastoma.

Glioblastoma multiforme (GBM) is the most malignant and multiple tumors of the central nervous system. The survival rate for GBM patients is less than 15 months. We aimed to uncover the potential mechanism of GBM in tumor microenvironment and provide several candidate biomarkers for GBM prognosis. In this study, ESTIMATE analysis was used to divide the GBM patients into high and low immune or stromal score groups. Microenvironment associated genes were filtered through differential analysis. Weighted gene co-expression network analysis (WGCNA) was performed to correlate the genes and clinical traits. The candidate genes' functions were annotated by enrichment analyses. The potential prognostic biomarkers were assessed by survival analysis. We obtained 81 immune associated differentially expressed genes (DEGs) for subsequent WGCNA analysis. Ten out of these DEGs were significantly associated with targeted molecular therapy of GBM patients. Three genes (S100A4, FCGR2B, and BIRC3) out of these genes were associated with overall survival and the independent test set testified the result. Here, we obtained three crucial genes that had good prognostic efficacy of GBM and may help to improve the prognostic prediction of GBM.

[Identification of A Case of DEL Blood Group with DAT Positive Phenotype and Pedigree Analysis].

OBJECTIVE: To identify the blood group of a patient with DEL phenotype combined with positive direct anti-human globulin test and to analyze the pedigree. METHODS: Routine serological reagents were used for serological analysis of RhD blood group in the pedigree members. Exons and flanking sequences of RHD gene were amplified, sequenced and analyzed for heterozygosity. The familial genetic state of DEL phenotype was further analyze in the family members. RESULTS: The DAT was strongly positive in the proband. The 1227G>A allele (RHD*DEL1) was present in the exon 9 of RHD gene, and the mother was the carrier of RHD*DEL1. The proband was identified as RHD+/RHD-, suggesting the CD1227Ae/Cde haplotype. CONCLUSION: The proband is DEL phenotype (RHD*DEL1).

Draft Genome Sequence of the Histamine-Producing Bacterium Enterobacter kobei Strain 42-12.

Bacterially produced histamine in food can be a cause of food poisoning. The whole-genome sequence is described for one histamine-producing Enterobacter kobei 42-12 isolate from the edible portion of salted, dried fish.

[Identification of Phenotype and Genotype in One Case with Weak D blood group].

OBJECTIVE: To investigate the phenotype and genotype of the weak D blood group in one case of Chinese Han people. METHODS: Phenotype of blood sample was identified with serologic tests; PCR-SBT was applied for the analysis of genotype and RhD zygosity. RESULTS: Both saline and gel card tests demonstrated this case to be dCcee, which was contrary to glass bead card result. Some of the RBC D epitopes were negative.c.1022T>A allele was identified with PCR-SBT and the zygosity analysis showed this case to be D/d. CONCLUSION: RHD*1022 A is more suitable to be categorized as weak partial D other than weak D in a Chinese Han people.

Characterization of microRNAs expression profiling in one group of Chinese urothelial cell carcinoma identified by Solexa sequencing.

OBJECTIVE: The goal of this study was to extend the known repertoire of microRNAs (miRNAs) expressed in urothelial bladder cancer (BCa) of the Chinese population and further understand the molecular events of miRNAs underlying urothelial bladder tumorigenesis at the global genome level. MATERIALS AND METHODS: We separated well-characterized epithelial tumor cells from 20 moderately differentiated or poorly differentiated BCa specimens by laser capture microdissection (LCM) and pooled these cells of interest prior to RNA analysis. Ten normal bladder epithelia (NBE) samples were pooled as the control. After preparation of small RNAs library, the 2 samples were sequenced simultaneously by the next generation high through-put Solexa sequencing technology. RESULTS: We employed the next generation high through-put Solexa sequencing technology to clone and identify miRNAs in BCa and NBE, and generated 11,146,610, and 10,263,845 high quality sequence reads, respectively. According to the analysis of size distribution, 22 nt class was the most abundant group of small RNAs in the BCa. Likewise, the 20 and 22 nt sequences were significantly greater than shorter or longer sequences, and accounted for 59.55% of the total sequence number of NBE library. The whole-genome-scale data mining suggested that BCa and NBE libraries both contained multiple and heterogeneous small RNA species. On further analysis, the sequencing data revealed that different miRNAs showed clearly in-house differential expression levels in BCa and NBE and 74 miRNAs aberrantly expressed between BCa and NBE at the global genome level. We also predicted 13 novel miRNAs in both BCa and NBE libraries. CONCLUSIONS: Our results suggest that BCa miRNAs include a large proportion of conserved miRNAs and a set of non-conserved miRNAs with low expression levels. These known and newly identified miRNAs at the population level significantly enhance our knowledge of BCa miRNAs expression profiling and provide insights into miRNAs oncogenesis and oncotherapy in BCa. Further studies are necessary to elucidate the roles of miRNAs in urothelial bladder tumorigenesis and determine the potential of miRNAs as diagnostic and prognostic tools or therapeutic targets for BCa in the future.

[Association between polymorphisms of inflammatory factor genes and coronary heart disease].

OBJECTIVE: To investigate the association between genetic polymorphisms of inflammatory factors and susceptibility to coronary heart disease(CHD) in southern Chinese Han population. METHODS: Using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) method, the genotypes of five inflammatory factors (BRCA1-associated protein, a disintegrin and metalloproteinase 8, inter-alpha-trypsin inhibitor H3, interleukin-15, cyclooxygenase-2) were anaylzed in 283 CHD patients diagnosed by angiography and 176 controls. RESULTS: In these inflammatory factors, the 270T/C and 90A/G polymorphisms of the BRAP gene showed a significant association with CHD. The allele and genotype frequencies of BRAP gene were consistent with those predicted by Hardy-Weinberg equilibrium (chi-square=0.878, P> 0.05; chi-square=0.776, P> 0.05, respectively). The frequencecies of 270C and 90G alleles in CHD patients was significantly higher than those of the control group (29.51% vs. 21.31%, P=0.006; 30.04% vs. 21.31%, P=0.004, respectively). Compared with 270TT and 90AA, 270CC and 90GG genotypes had a significantly increased CHD risk by Logistic regression analysis (OR=4.51, 95%CI: 1.41-14.45, P=0.011; OR=5.09, 95%CI: 1.60-16.26, P=0.006, respectively). This association was still signifcant after adjustment for the sex, age, smoke, hypertension, diabetes, plasma total cholesterol and low density lipoprotein levels. No evidence of association was found for other single nucleotide polymorphisms. CONCLUSION: The 270T/C and 90A/G polymorphisms in the BRAP gene may contribute to an increased risk of CHD among southern Chinese Han population.

[Study on the correlation between CETP TaqIB, KCNE1 S38G and eNOS T-786C gene polymorphisms for predisposition and non-valvular atrial fibrillation].

OBJECTIVE: To study whether CETP TaqIB,KCNE1 S38G and eNOS T-786C genetic polymorphisms are associated with non-valvular atrial fibrillation in the Han population from Zhejiang province. METHODS: Polymerase chain reaction restriction fragment length polymorphism assay was used to detect the distribution of alleles and genotypes of CETP TaqIB, KCNE1 S38G and eNOS T-786C in 147 patients with non-valvular atrial fibrillation and in 147 subjects as controls in Han population of Zhejiang province. RESULTS: (1) The frequency of CETP B1 allele in NVAF patients was higher than that of the control group and showing a statistically significant difference (OR = 1.763, 95% CI: 1.247-2.492, P = 0.002). (2) Results from logistic regression analysis revealed that: after adjustment of confounding variables such as sex, age, smoking, hypertension and body mass index, data from the binary logistic analysis showed a statistically significant difference in CETP TaqIB genetic polymorphism between patients and controls. (3) From multifactor dimensionality reduction analysis, results showed an interaction of CETP TaqIB, KCNE1 S38G and eNOS T-786C genetic polymorphisms. Odds ratio of the three simultaneously existing genetic polymorphisms was 1.849 times more than CETP TaqIB alone. CONCLUSION: CETP BI allele was an independent risk factor for predisposition to non-valvular atrial fibrillation. These findings suggested that the simultaneous existence of CETP B1, KCNE1 S38G and eNOS T-786C allele might be elevated with the predisposition to non-valvular atrial fibrillation in the Han population of Zhejiang province.

[Changes of platelet aggregation function of apheresis collected platelets and soluble P-selectin during storage].

The objective of this study was to explore the changes of aggregation function of apheresis platelets and soluble P-selectin (sP-selectin) during storage. 20 samples of apheresis platelets were collected, and the aggregation function were examined by function test and the level of sP-selectin every day in storage of 5 days. The results showed that the aggregation function of platelets declined obviously during storage, there were significant differences between the first-day group and any of the other groups (p < 0.01). The max platelet aggregation rate was < or = 3% in the fourth-day group; sP-selectin level in plasma increased with prolong of storage time; there were significant differences between the first-day group and any of the other groups (p < 0.05). In conclusion, platelets were activated continuously during storage, while its aggregation function declines significantly. The ability of platelet aggregation to response to ADP loses almost completely since the fourth day during platelet storage. It should be paid more attention to the damage of apheresis collected platelets during storage.

[Frequency and transcript variant analysis of gene fusions between TMPRSS2 and ETS transcription factor genes in prostate cancer].

OBJECTIVE: To investigate the frequencies and types of fusions between the transmembrane protease serine 2 (TMPRSS2), ETS-related gene (ERG), ETS variant-1 (ETV1), and ETS variant-4 (ETV4) genes in prostate cancer (PCa) and significance thereof. METHODS: Biopsy samples of prostate were obtained under transrectal ultrasound (TRUS) from 32 PCa patients, aged (74 +/- 8) and 34 patients with benign prostate hyperplasia (BPH). Nested RT-PCR and direct DNA sequencing were used to detect the fusion genes of TMPRSS2/ERG, TMPRSS2/ETV1, and TMPRSS2/ETV4. The association between the fusion-positive tumor rate and Gleason grading was analyzed. RESULTS: Of the 32 PCa patients, TMPRSS2/ERG fusion was detected in 17 cases (53.1%), including 5 variant fusion transcripts one of which was newly discovered with the GenBank accession number of EU090248. TMPRSS2/ETV1 fusion was detected in only 2 cases (6.3%), including one newly discovered variant fusion transcripts with the GenBank accession number of EU090249. TMPRSS2/ETV4 fusion was not detected. The positive rates of TMPRSS2/ERG and TMPRSS2/ETV1 fusions showed no statistical association with the Gleason grade (P = 0.169). No fusion between the TMPRSS2 and ETS transcription factor genes was detected in the 34 BPH samples. CONCLUSION: TMPRSS2/ERG and TMPRS22/ETV1 fusion genes with different subtypes exist in the tissues of PCa. TMPRSS2/ERG and TMPRSS2/ETV1 fusion genes may be used as diagnostic tools for PCa.

[Detection of urine DD3/PSA mRNA ratio by duplex TaqMan RT-PCR assay and evolution of its primary application].

OBJECTIVE: To establish a duplex TaqMan RT-PCR assay for urine DD3/PSA mRNA ratio, and to evaluate its effect in diagnosis of prostate cancer (PCa). METHODS: Urine samples were obtained from 34 patients with PCa and 44 patients with benign prostate hypertrophy (BPH) by prostate massage. A duplex TaqMan RT-PCR assay was developed: PCR primers were designed to amplify the fragments between the exon1 and exon2 in the PSA mRNA and between the exon1 and exon3 in the DD3 mRNA, and the PCR TaqMan-MGB probes were labeled with HEX and FAM respectively in 5' for PSA mRNA. LNCaP cells were used as template. DD3/PSA mRNA ratio was measured. Receiver operating characteristic curve (ROC) was drawn so as to evaluate its diagnostic efficacy. RESULTS: Sequencing showed that the PCR products were specific for PSA mRNA and DD3 mRNA. The minimum detection level was approximately 0.6 cells/reaction for PSA mRNA and was 60 cells/reaction for DD3 mRNA in the LNCaP cell cDNA. The intra- and inter-assay coefficients of variation of DD3/PSA mRNA were 3.8%-4.7% and 4.1%-4.9% respectively. Urine DD3/PSA mRNA ratio in PCa group was significantly higher than the BPH group (P < 0.01). When the cutoff value was defined as 0.254, the area under curve (AUC) of DD3/PSA mRNA ratio was 0.746 (95% CI: 0.630-0.862), and the sensitivity and specificity were 64.7% and 77.3% respectively. The urine DD3/PSA mRNA ratio positive rate was not correlated with clinical and pathological parameters. CONCLUSION: A duplex TaqMan RT-PCR assay for urine DD3/PSA mRNA ratio has been established with an excellent clinical performance and specificity for PCa, saving time and reducing costs. It may be a promising method in the early diagnosis of PCa.

[Quantitative detection of DD3 mRNA in prostate cancer tissues by real-time fluorescent quantitative reverse transcription polymerase chain reaction].

OBJECTIVE: To analyze the expression of DD3 mRNA in the prostate tissues. METHODS: DD3 mRNA was detected by realtime fluorescent quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR) based on the Taqman technique in the tissues of 27 patients with non-prostate cancer( NPCa), 21 prostate cancer( PCa), 39 benign prostatic hyperplasia (BPH) and 15 normal prostate (NP). The ROC curve was used to evaluate the diagnostic value of DD3 mRNA. RESULTS: DD3 mRNA expression was not detected in the NPCa tissues. The median expressions of DD3 mRNA in PCa, BPH and NP tissues were 7. 2 x 10(6), 2. 5 x 10(4) and 1.5 x 10(4) copies/mg tissue, respectively. The DD3 mRNA expression levels were significantly different between nonmalignant and malignant tissues (P < 0.01). No significant differences in DD3 mRNA expression were detected between the NP and BPH tissues and no significant correlation was found between the DD3 mRNA expression and clinical pathological parameters. The AUC-ROC was 0.937 (95% CI: 0.879 - 0.995) at cutoff value 1.4 x 10(5) copies/mg tissue. The sensitivity, specificity, accuracy, positive predictive value, negative predictive value, positive likelihood ratio and negative likelihood ratio for DD3 were 90.5%, 85.0%, 86.7%, 76.0%, 94.3%, 6.03 and 0.11 respectively. CONCLUSION: The DD3 mRNA expression is confined to prostate tissues and highly upregulated in PCa tissues. It has a potential application value in the early diagnosis of prostate cancer and the follow-up of the patient.

[Quantitative detection of differential display code 3 mRNA in peripheral blood of patients with prostate cancer].

OBJECTIVE: To investigate the expression of DD3 mRNA in the peripheral blood and its value in diagnostic of prostate cancer (Pca). METHODS: Thirty-five untreated Pca patients, aged 72 (53 - 83), 58 Pca patients treated with endocrinotherapy, aged 74 (53 - 86), and 59 patients with benign prostatic hyperplasia, aged 71 (48 - 85), underwent peripheral blood sample collection one week before or after digital examination. RT-PCR was used to examine the level of DD3 mRNA. The level of prostate specific antigen (PSA) was detected too. Ten healthy male frontiers, aged 33 (21 - 38), were used as controls. Receiver operating curve (ROC) was drawn to evaluate the diagnostic performance of DD3 mRNA. RESULTS: The DD3 mRNA level of the untreated Pca patients was 2741 copies/ml, significantly higher than those of the Pca patients treated with endocrinotherapy, patients with benign hyperplasia, and healthy persons (all < 24 copies, all P < 0.001). The DD3 mRNA level of the endocrinotherapy-treated Pca patients was significantly higher than those of the patients with benign hyperplasia, and healthy persons (both P < 0.001). ROC analysis showed that when the critical value was 846 copies/ml the area under curve of ROC (AUC-ROC) was 0.8233 (95% CI: 0.725 - 0.910). and the sensitivity was 74.34%, the specificity was 89.8%. The DD3 mRNA in the peripheral blood increased along with the increase of clinical staging (P < 0.01). CONCLUSION: The level of DD3 mRNA in the peripheral blood is an excellent marker for diagnosis pf Pca, and may help in monitoring of the curative effects of endocrinotherapy.

[The effects of lupus anticoagulant on pulmonary thromboembolism].

OBJECTIVE: To evaluate the effects of lupus anticoagulant (LA) on pulmonary thromboembolism (PTE). METHODS: Thirty-eight patients with PTE (17 massive and 21 submassive) and 30 healthy adults were studied. Russell's viper venom time (RVVT) was used to examine the ratio of LA (LAR), and a colorimetric method was used to detect the activity of plasma protein C (PC:A) and radioimmunoassay (RIA) was employed to measure the level of plasma thromboxane B(2) (TXB(2)) and 6-keto-prostaglandin F1alpha (6-keto-PGF1alpha). RESULTS: Compared with the normal group, LAR, TXB(2) and TXB(2)/6-keto-PGF1alpha showed significant increase in the massive PTE and the submassive PTE groups (P < 0.01), and the levels were higher in the massive group than in the submassive group (P < 0.01). Both groups showed significant decrease in PC:A and 6-keto-PGF1alpha compared with the normal group (P < 0.01). CONCLUSIONS: LA can increase TXB(2)/6-keto-PGF1alpha and decrease PC:A in patients with PTE. It is suggested that there may be an association between the increase of LAR and the presence of PTE.