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Objective Next Generation Sequencing(NGS) platform was used to study the characteristics of hot gene mutations in non-small cell lung cancer (NSCLC). The distribution, type and frequency of mutation sites were systematically analyzed to evaluate the pathogenicity of mutation sites . Methods A total of 94 NSCLC tissue samples were included in this study including paraffin-embedded (FFPE) samples and fresh tissue samples, which were collected from July 2015 to April 2017 at the Qilu Hospital of Shandong University. The patient's age ranged from 35 to 82 years with a median age of 61 years. There were 63 males and 31 females. 22 hot genes in NSCLC were selected as the detection panel, including KRAS, EGFR, BRAF, PIK3CA, AKT1, ERBB2, PTEN, NRAS, STK11, MAP2K1, ALK, DDR2, CTNNB1, MET, TP53, SMAD4, FBXW7, NOTCH1, ERBB4, FGFR1 and FGFR2. Mutation detection was performed using the Ion AmpliSeq Colon and Lung Cancer Panel of the Thermo fisher's Ion Torrent sequencing platform. The sequencing data was analyzed using Ion Torrent suite v4.4.2 software. Results Among the 22 mutant genes commonly found in NSCLC, the mutation frequency of TP53 was the highest, accounting for 46.9％ of all mutations, followed by the EGFR mutation (28.1％); A total of 89 mutations were detected, including 63 hot spot mutations (reported mutations) and 26 new mutations (unreported mutations). The most frequently detected mutation was the frameshift deletion of exon 19 of EGFR, followed by the mutation of exon L858R;Analysis of the mutation in targeted drug sites of EGFR showed that the frameshift deletion of exon 19 of EGFR was the most frequently detected, followed by the mutation of exon L858R on chromosome 21. Bioinformatics software was used to analyze the pathogenicity of 26 new mutation sites. Results showed that in addition to ATK1:c. 47-12G>A and TP53: c. 214 C>G, the remaining 24 new mutation sites had at least one major impact on the gene function in three aspects, including gene conservation, amino acid sequence change and protein structure influence. Conclusion In this study, NGS was used to conduct combined detection of mutation sites of multiple hot genes, which might cover more comprehensively genetic variation and provide a basis for screening the most suitable targeted therapy groups. The pathogenicity prediction of new mutations and the changes in tumor-related signaling pathways involved provide a reference for further study of the pathogenesis of NSCLC.
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Objective To evaluate the diagnostic value of CEA, CA19-9, CA72-4, AFP, and CA125 in gastric stromal tumors. Methods 41 patients with gastric stromal tumors and 11 patients with gastric leiomyoma were recruited in this study from Qilu Hospital of Shandong University during May 2014 to December 2017. The tissue was collected by surgery, and HE staining was done. Immunohistochemistry was employed to detect the expression of CD34, CD117, and DOG-1. Serum of all cases and 41 healthy volunteers in the same hospital were collected. The levels of CEA, CA19-9, CA72-4, AFP and CA125 were examined by electrochemiluminescence assay, and the differences in each group were compared by M-W test or K-W test. Then the relationship between those biomarkers and the clinical parameters of gastric stromal tumors was analyzed. Moreover, AUC (Area Under the curve), cut-off value, sensitivity, specificity, positive predictive value, negative predictive value with drawing ROC curve (receiver operating characteristic curve) were also calculated. Results Spindle cells or epithelioid cells were observed in the tissue of gastric stromal tumors. The expression of CD34, CD117, DOG-1 were positive. The level of the serum CEA 1.53 (1.15, 2.22) ng/ml in patients with gastric stromal tumor patients was higher than that in healthy controls 1.06 (0.62, 1.48) ng/ml and that in patients with gastric leiomyoma 0.79 (0.39, 1.39) ng/ml (the U value was 446.5, and 113.0 respectively, P<0.05). The level of CA19-9 in gastric stromal tumors 9.30 (4.95, 12.70) U/ml was higher than that in healthy controls 6.62 (4.56, 8.82) U/ml (the U value was 615.5, P<0.05). There were no significant differences in AFP, CA125, and CA72-4 of three groups (the H value was 4.348, 1.073, and 3.897, P>0.05). Furthermore, the level of CEA was closely related to TNM stage (the U value was 129.0, P<0.05). The diagnostic value of CEA and CA19-9 was statistically not significant (P>0.05). However, the combination of two markers might increase diagnostic efficiency. The sensitivity, specificity, positive predictive value, negative predictive value and AUC was 92.7％, 48.8％, 64.4％, 87.0％ and 0.752 respectively. Conclusion The combination of CEA and CA19-9 has better sensitivity and negative predictive value in auxiliary diagnosis of gastric stromal tumors.
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Objective To investigate relationships between signal/cutoff (S/CO) ratios of antiHCV recombinant immunoblot assay (RIBA) and their positivity with different chemiluminescence immunoassay(CLIA) reagents.Methods A case-control study was performed.From March 2014 to March 2015,anti-HCV antibody was detected in 2 616 serum of outpatients and inpatients coming from Department of Clinical Laboratory,Qilu Hospital of Shandong University by three kinds of homemade CLIA reagents and one imported CLIA reagents.The positive samples were further tested by RIBA.The correlation between the positivity and the S/CO ratios was analyzed.The difference between different reagents were compared by x2 method.Results The predicted positivities of Shandong Laibo were 97.8％ and 33.3％ with S/CO ratio ≥ 26.8 and 1 to 26.8,respectively;The predicted positivities of Beijing Yuande were 96.7％ and 20％ with S/CO ratio ≥ 16.6 and 1 to 16.6,respectively;The predicted positivities of Beijing Kemei were 97.0％ and 9.8％ with S/CO ratio ≥ 16.7 and 1 to16.7,respectively;The predicted positivities of Abbott were 96.9％ and 12.8％ with S/CO ratio≥5 and 1 to 5,respectively.Conclusions Anti-HCV CLIA S/CO ratio and RIBA confirmatory test results have some relevance.Domestic reagents also can refer to import reagents determine the relationship between the positivity and the S/CO ratio.Different domestic reagent of positivity has different S/CO ratio.Although each reagent S/CO ratio to the same positivity has large difference,suggesting each manufacturer should set their products corresponding values according to the situation,providing reference for the clinical use of unit in result determination of the clinical trials.
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Objective To study the characteristics of IL-1B-31/-511 single nucleotide polymor-phisms (SNPs) and the variable number tandem repeat (VNTR) in intron 2 of the IL-1ra gene (IL-1RN) in patients with HCV-related liver diseases .Methods The concentration of IL-1βand IL-1ra in serum sam-ples was measured by ELISA assay .The SNPs of IL-1B gene (-31C/T,-511C/T) from 310 cases with HCV infection and 324 unrelated healthy controls were determined by using gene chip analysis , and the results for some randomly selected specimens were compared with those by using polymerase chain reaction -restriction fragment length polymorphism ( PCR-RFLP) assay.The VNTR polymorphism of IL-1RN intron 2 was ana-lyzed by PCR-RFLP assay.The serum level of alanine aminotransferase (ALT), an indicator of hepatocellu-lar injury, was detected by ROCHE cobas 8000 analyzer.HCV replication was measured by using specific fluorescence PCR .The genotypes of HCV were determined by direct nucleotide sequencing test .Results Compared with control group, the serum level of both IL-1β[(22.6 ±7.3) vs (13.7 ±4.2)] pg/ml and IL-1ra [(286.30 ±55.10) vs (185.55 ±48.32)] pg/ml were significantly increased in patients with HCV infection ( P0.05).The frequency of IL-1B-511TT genotype (P<0.05, OR=1.55, 95% CI =1.10-2.18) and IL-1B-511T allele (P<0.05,OR=1.31,95% CI=1.05-1.63) in patients with HCV infection were signifi-cantly higher than those in healthy controls .IL-1B-511C/T SNP showed a significant association with the outcomes of HCV infection (P<0.005).Compared with IL-1B-511CC and IL-1B-511CT, IL-1B-511TT was a major risk factor for mild and moderate Hepatitis C [ OR=2.17 ( 1.48-3.19 ) ] , severe Hepatitis C [OR=2.11(1.05-4.26)], cirrhosis [OR=2.98(1.77-4.99)] and HCC [4.33(2.16-8.67)].IL-1B-511 T allele was significantly associated with mild and moderate Hepatitis C [ 1.80 ( 1.38-2.36 ) ] , severe Hepatitis C [1.80(1.08-3.01)], cirrhosis [2.62(1.76-3.89)] and HCC [3.49(1.96-6.23)].The fre-quency of IL-1B-511T allele showed significant difference among each group (P<0.005).No association was found between any of the other polymorphisms and HCV infection .Conclusion The serum level of IL-1βand IL-1ra were significantly associated with HCV infection .IL-1B-511T allele in patients with HCV in-fection up-regulated the serum level of IL-1β.IL-1B-511TT and IL-1B-511T allele were major risk factors for mild and moderate Hepatitis C, severe Hepatitis C, cirrhosis and HCC, but IL-1B-511CC/C had oppo-site effects.
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OBJECTIVE To investigate the incidence of bacteria colonization on lumbar epidural catheter tips in postoperative analgesia patients. METHODS The catheter tips were cultured in 100 patients with ASA grade Ⅰ-Ⅱ undergoing lower extremity osteoarticular operation.Lumbar epidural catheters were placed in the operating room with aseptic technique.Diluted local anesthetic and fentanyl infusions were used for postoperative analgesia.The epidural catheter was removed with aseptic technique and the tips sent for microbiological culture after 3 days. RESULTS From 100 patients,bacteriological examination revealed bacteria colonization in 9(9.0%),mainly Staphylococcus epidermidis(6;66.7%),followed by Enterococcus(1;11.1%),Gram-negative bacilli(1;11.1%),and yeasts(1;11.1%).No patient developd infectious complications. CONCLUSIONS Our results show that the risk of bacteria colonization associated with lumber postoperative analgesia in three days is low.No patient develops local or central nervous infection.Epidural postoperative analgesia can be routinely used without worry of infection in epidural space.But we recommend prophylactic measures should be applied in the high-risk groups.
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0 05). In 53 ITP patients, the positive rate of PAIgG was 64 2% (34/53), the positive rate of mean fluorescence intensity (MFI) was 71 7% (38/53), the positive rate of both PAIgG and MFI was 84 9% (45/53). The expression of GPⅡb/Ⅲa in ITP group without clinical symptoms was significantly higher than that in ITP group with clinical symptoms ( P