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Immune checkpoint inhibitors (ICIs) targeting CTLA-4 and PD-1 /PD-L1 are increasingly used to treat several types of cancer.Although several agents have been approved,only 10％ to 30％ of patients have benefited from them.Some studies have shown that intestinal micro-ecology can affect the outcome of ICIs through immune regulation in cancer patients.This literature review s summarized the regulation and predictive functions of intestinal micro-ecology on ICIs treatment,and introduced possible mechanisms of intestinal micro-ecology that affacted the efficacy of immunotherapy.In addition,some approaches for detecting gut microbiome were also summarized.Gut microbiota highlighted in this review may serve as novel biomarkers to predict clinical outcomes in cancer patients.
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Objective@#To establish a diagnostic prediction model for esophageal squamous cell carcinoma (ESCC) and search the potential biomarkers of ESCC. @*Methods@#Serum samples from 59 patients with ESCC and 57 healthy controls were collected, and randomly divided into the training group (44 patients and 42 healthy controls) and validation group (15 patients and 15 healthy controls). Serum proteins/peptides were extracted and purified with weak cation-exchange chromatography Magnetic Beads (WCX-MB), and detected by the matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). Then the differentially expressed proteins/peptides were screened out, and a diagnostic prediction model for ESCC was established and preliminarily validated. @*Results@#The ClinProTools software identified 31 differential peptide peaks (P＜0.05), among which 18 peaks had significant difference (P＜0.01). Compared with healthy controls, 8 peaks were up-regulated in ESCC patients, while 10 peaks were down-regulated. Among them, the areas under the receiver operating characteristics (ROC) curve (AUC ROC ) of m/z 2 660.84 and m/z 5 336.49 peaks were 0.95 and 0.91, respectively, and their expressions were up-regulated in ESCC patients. The validation results showed that the accuracy, sensitivity and specificity of the diagnostic prediction model established by the genetic algorithm (GA) were 93.10%, 92.90% and 93.30%, respectively. @*Conclusion@#The established diagnostic prediction model may be used for the auxiliary diagnosis of ESCC. Two peptide peaks of m/z 2 660.84 and m/z 5 336.49 may be the potential biomarkers of ESCC.
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Objective To explore the significance of long non-coding RNA (lncRNA) taurine up-regulated gene 1 (TUG1) in hepatocellular carcinoma (HCC),to predict the target gene of TUG1,and to provide a ref-erence for further study of TUG1 in HCC.Methods The differential expression of TUG1 in HCC was ana-lyzed by using the UALCAN database and the survival analysis of TUG1 was performed.The target gene of TUG1 was predicted by RegRNA 2.0 biology software,HMDD,targetscan and microT-CDS,and the regulato-ry network of lncRNA TUG1-microRNAs-mRNAs was constructed.The predicted target gene was analyzed by Gene Ontology (GO) and KEGG signal transduction pathway enrichment by using FunRich platform. Results TUG1 expression in HCC was significantly increased,and the expression level of TUG1 increased generally with the increase of tumor grade.The overall survival of patients with low expression of lncRNA TUG1 was significantly longer than that of lncRNA TUG1 high expression patients.There were four possible binding sites of HCC related microRNAs (hsa-mir-122-5p,hsa-mir-200a-3p,hsa-mir-34c-3p,hsa-mir-629-3p) on TUG1,which regulated 245 downstream target genes and formed the regulatory network of lncRNA TUG1-microRNAs-mRNAs.In the biological process,microRNA target genes were highly enriched in the processes such as the regulation of nucleobase,nucleoside,nucleotide and nucleic acid metabolism.In KEGG pathway analysis,microRNA target genes were highly enriched to the signal pathways mediated by Syndecan and TRAIL.Conclusion TUG1 expression level in HCC increased.Increased expression of TUG1 is associat-ed with poor prognosis in HCC.Bioinformatics methods can be used to explore the mechanism of tumorigene-sis from the molecular level,which can provide valuable information for subsequent experiments and clinical diagnosis and treatment.
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Objective To use the liquid protein combined with MALDI-TOF-MS for screening the serum differential peptides markers in lung adenocarcinoma patients and to establish the lung adenocarcinoma diag-nosed prediction model for founding the potential markers for the diagnosis of lung adenocarcinoma.Methods 37 patients with lung adenocarcinoma and 33 healthy subjects and benign lung disease which were made up in control group were collected,in the two groups the age and the sex were matched.The two groups were ran-domly divided into training group(30 cases of lung adenocarcinoma,26 cases of control)and test group(7 ca-ses of lung adenocarcinoma,7 cases of control)according to 3:1.T he differential diagnosis of lung adenocarci-noma and control group was performed by liquid chip-time-of-flight mass spectrometry and software ClinPro-Tools 3.0 to establish a prediction model of lung adenocarcinoma.The diagnostic model was validated by using serum samples from the test group to assess the diagnostic efficacy of the model.Results Nine peptide peaks with significant differences(P<0.05)were obtained by ClinProTools 3.0 software analysis.The up-regulated peaks in lung adenocarcinoma(m/z)were 8 976.5,4 469.05,4 966.78,8 925.5,4 531.05,and the down-reg-ulated m/z were 3 304.44,8 594.76,3 266.82,3 195.52.According to the genetic algorithm(GA),the lung ad-enocarcinoma diagnosis and prediction model was established.The overall recognition ability of the model was 94.49%.The model was evaluated by the test group.The results showed that the sensitivity of the model was 100.0% and the specificity was 85.7%.Conclusion Among lung adenocarcinoma patients,serum benign lung disease and healthy,there are differences in the serum peptide.T he use of differential peptide peaks to estab-lish lung adenocarcinoma diagnostic prediction model for the early diagnosis of lung adenocarcinoma provides a new method.
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Objective To prepare the mAbs against hK6 for establishing a sandwich enzyme-linked immunosorbent assay(ELISA) of hK6,and exploring its clinical value.Methods The hybridoma technique was used to prepare mAbs against hK6.The mAbs were purified and labelled with horseradish peroxidase for the sandwich ELISA method.The sandwich ELISA method was used to detect the serum hK6 concentrations in patients with malignant gastric neoplasm.Then the best antibody pair was selected from coating antibody and enzyme-linked antibody to establish a sandwich ELISA method through the chessboard titrations.Compared with CEA,we explored the feasibility of hK6 as gastric cancer biomarkers.Results A sandwich ELISA method was established for quantifying hK6 in serum.The results showed that the optimal concentration of coating antibody was 5 μg/mL.The optimal concentration of enzyme-linked antibody was 1:2 000.Serum hK6 in the patients with gastric cancer groups[(5.78±1.66)ng/mL] than healthy individuals groups[(3.35±0.67)ng/mL] and those in gastric ulcer groups[(3.59±1.02)ng/mL],the difference was statistically significant(P0.05).The hK6 positive rate of gastric cancer was 69.70％,and CEA was 45.46％.In the combined detection,the positive rate was 78.79％.Conclusion A sandwich ELISA is established successfully.As a favorable serum biomarker for gastric cancer,the detection of hK6 together with CEA is helpful in the diagnosis of gastric cancer.
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Objective To investigate the clinical distribution and antimicrobial resistant characteristics of Pseudomonas aeruginosa(PA)in the northern area of Guangdong.Provide the reference for clinical to prevent infection and reasonable choice of antibiotics and reduce the production of drug resistance strains.Methods The separation and identification of PA were performed by conventional methods during the data of drug 2013 and 2014.The data of sensitivity test of PA were analyzed by WHONET 5.6 and SPSS19.0 softwares.Results The 584 strains PA were mainly distributed in ICU,department of orthopaedics and respiratory medicine.Specimens were mainly from sputum and wound secretion.The detection of PA to 12 antibacterial agents showed different resistance.The antimicrobial with highest resistance was the gentamicin and lower resistance rates to fluoroquinolones,carbapenems,enzyme inhibitors.And a downward trend was shown in drug resistance to CIP,FEP,LEV,SCF.Conclusion PA mainly cause lung and wound infection,especially those old patients that come from ICU,department of orthopaedics and respiratory medicine.Although the drug resistance rates of PA to the commonly used antibiotics are relatively low,The clinicians should reasonably use antibiotics so as to reduce the resistant strains,especially the produce of MDR-PA and PDR-PA.
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Objective To study comparatively the performance of rapid plasma reagin (RPR) test in syphilis detection among pregnant women and non-pregnant women to provide reference for detecting syphilis in pregnant women.Methods The women aged 20-40 years old were selected and divided into the pregnant group and the non-pregnant group.RPR and treponema pallidum particle assay(TPPA) were simultaneously adopted to conduct the syphilis detection.The positive cases were judged by the TPPA detection results combined with the contact history,clinical symptoms and treatment situation.The results were compared with those by RPR for determining the false negative and false positive in RPR.The false negative rate and false positive rate of RPR detection results were analyzed in the two groups.Results Among 117 pregnant women,15 cases were false negative in RPR and 9 cases were false positive in RPR;among 755 non-pregnant women,there were 44 cases of false negative RPR results and 8 cases of false positive RPR results.The false negative rates in the pregnant group and non-pregnant group were 25.0% and 8.8% respectively,the difference was statistically significant(χ2=14.739,P<0.05);the false positive rates in the pregnant group and non-pregnant group were 15.7% and 3.1% respectively,the difference was statistically significant (χ2=14.722,P<0.05).Conclusion There are many factors affecting RPR for detection syphilis,pregnant women are the specific group,so higher false positive rate and false negative rate exist than non-pregnant women,the detection results should be comprehensively judged by combining with clinical symptoms and disease history,if necessary,combining with other syphilis detection method for avoiding missed diagnosis and misdiagnosis.
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Objective To establish a rapid diagnostic method for the detection of marine vibrios,and then construct a new technology platform for the clinical diagnosis of marine vibrio infection.Methods A pair of PCR primers and a sequencing primer based on the whA gene of V.vulnificus and the toxR genes of V.parahemolyticus and V.alginolyticus were designed respectively,and then the specific DNA fragments were amplified.Next,the single-stranded DNA templates were prepared for pyrosequencing.The obtained base sequence was validated by NCBI alignment.In addition,the 16S rRNA genotyping of V.vulnificus was also performed.Results The PCR primers and sequencing primer of V.vulnificus showed good specificity,and a 167 bp DNA fragment was amplified from 4 strains of V.vulnificus.The pyrosequencing results completely matched with the whA gene sequence of V.vulnificus.Meanwhile,the control strains were negative.A 105 bp DNA fragment and a 134 bp DNA fragment were amplified from 11 strains of V.parahemolyticus and V.alginolyticus,respectively,and the pyrosequencing results were consistent with the expected sequence.In addition,one of 4 strains of V.vulnificus was identified as 16S rRNA-A type,and the other 3 as 16S rRNA-B type.Conclusion The PCR-pyrosequencing method established in this study is a new method for the real-time detection of short nucleotide sequences.It has some advantages such as high throughput,high precision and simple operation,and may be applied to the fast and accurate identification of marine and terrestrial pathogenic bacteria.
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Objective To investigate the distribution and resistant pattern of Enterobacter cloacae to antimicrobial agents for reasonable use of antibiotics .Methods E .cloacae strains were isolated from patients from January 2009 to December 2013 .The strains were identified by VITEK‐2 Compact System of BioMerieux and tested for antimicrobial susceptibility by Kirby‐Bauer disk diffusion method .The results were analyzed .Results A total of 397 nonduplicate E .cloacae strains were isolated during 5 years ,accounting for 5 % of the total gram‐negative isolates . The strains were mainly isolated from sputum (48 .9% ) , followed by secretions (30 .5% )and urine (7 .3% ) .The percentage of E .cloacae strains resistant to all the antibiotics tested was on decline except carbapenems ,cefoxitin and amoxicillin‐clavulanic acid .Carbapenems were relatively more active against E .cloacae strains .The E .cloacae strains showed higher resistance rate toβ‐lactams .Conclusions It is necessary to strengthen the monitoring of clinical isolates to guide rational use of antimicrobial agents .
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The human tissue kallikrein(KLK)gene family consists of 15 highly conservative serine pro-teases,which is the largest uninterrupted cluster of protease genes in the human genome. Several members of the family are expected to be markers for tumor diagnosis and prognosis. Studies find that the expressions of KLK2-11 and KLK13-15 are abnormal,and the majority of KLKs have potential diagnostic and prognostic values.
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Objective To evaluate the ability of Vitek 2 Compact YST identification cards and pyrosequencing analysis for identifying the clinical isolates of yeast-like fungi.Methods Vitek 2 Compact YST identification cams and ITS1 region pyrosequencing analysis were used to identify the clinical isolates of yeast-like fungi at Jinan Military General Hospital in 2011.The strains which could not be identified to species by pyrosequencing analysis were identified again by the method of ITS1 region Sanger sequencing.The strains with inconsistent identified results by YST and pyrosequencing were identified again using API 20C.Results A total of 282 srtains were isolated from various clinical specimens for culturing in 2011.In addition to the three which could only be identified to similar species,other strains could be clearly identified to species by the method of ITS1 reverse pyrosequencing.Three strains which failed identified to species by pyrosequencing couldn't be distinguished fully by the method of Sanger sequencing.There were 7 strains unidentified,14 strains low discrimination(％ id ＜ 80％),and 6 strains misidentification using YST cams.The identification coincidence rate was 90.4％.The identification coincidence rates were various in different species of yeast-like fungi using YST cards.Conclusions The vast majority of clinical isolates of yeast-like fungi could be identified by ITS1 reverse pyrosequencing analysis.Vitek 2 Compact YST caRD basically meets the needs.But attention should be paid to the standard operating and regular internal quality control.For some rare species it is also needed to combine the source of specimen,colonies and characteristics to determine the identification results for the rare stains.
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Objective To investigate the characteristics of IL-22 in Helicobacter pylori (H.pylori)infection.Methods Thirty H.pyloripositive and fifteen H.pylori negative gastric biopsy specimens were enrolled,IL-22 mRNA expression was detected by real-time PCR and the protein level of IL-22 was determined by ELISA in gastric tissue.The H.pyloriand cell coculture system was established and IL-22 expression was measured by real-time PCR to investigate the main source of IL-22 in gastric tissue.The IL-22-producing T cell was examined by FCM in the gastric mucosa tissue.Results Gastric mucosal IL-22 mRNA and protein levels were significantly higher in H.pylori-positive patients than uninfected patients (P＜0.05).A H.pyloriand cell coculture system was established successfully and gastric epithelial cell and T cell were the main source of IL-22 in gastric tissue.IL-22 was produced by CD4+ and CD8+ T cells and these T cells were increased in H.pylori-infected gastric mucosa (P＜0.05).Conclusion IL-22 took part in H.pyloriinfection induced immune response and increased IL-22-producing T cells was the important feature of H.pyloriinfection.
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Objective To develop a real-time fluorescence quantitative PCR(FQ-RCR)method for detection of Kallkreins(KLK)4 gene expression in human breast cancer,and to investigate KLK4 expression levels in breast cancer and normal tissues.Methods KLK4 expression levels of 25 normal breast tissues and 39 cancer tissues were detected by the developed real-time quantitative PCR method.The statistical analysis for the relationship between KLK4 expression and several pathological parameters was performed by t test.Results The levels of KLK4 mRNA in normal breast and breast cancer tissue were 0.0120?0.0044 and 0.0272?0.0067 respectively(P0.05).Conclusions The level of KLK4 mRNA in breast cancer tissue was higher significantly than that in normal breast tissue.The results indicated that KLK4 gene expression may have relevance with breast cancer development but have no significant relevance with ER,PR,CerbB-2 and tumor metastasis.
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Objective To investigate the significance of IL-18R? expression on CD4+ T cells in peripheral blood (PB) of children with Mycoplasma pneumoniae pneumonia (MPP).Methods T cell subtype CD3/CD4/CD8 and expression of IL-18R? on CD4+ T cells in PB from 35 children with MPP and 15 age- and sex-matched control subjects was determined by flow cytometry.Results Compared with that of healthy control, CD3+ and CD4+ positive cells in MPP children were decreased (P0.05).Conclusion There exists cell-mediated immune function disorders and Th1/Th2 imbalance in children with MPP. Th1 immune response is dominant in acute-stage MPP.
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Expression levels of TGF-β1 mRNA in renal cortex of control group, diabetic group and taurine group measured by real-time quantitative RT-PCR were (7.0±0.8)×10 -3, (64.4±8.0)×10 -3 and (16.7±2.0)×10 -3, respectively. The corresponding results with end-point RT-PCR method were 0.28±0.12,0.58±0.16 and 0.43±0.10, respectively. Compared with end-point RT-PCR method, real-time quantitative with SYBR Green I was easier, faster, sensitive and specific.
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Objective To develop a real-time quantitative PCR method for detection of human breast cancer related novel gene-kallikrein gene 6 (klk6) expression and investigate klk6 expression levels in breast cancer tissue.Methods Using Sybr Green I, with GAPDH as reference, a real-time quantitative PCR method was established. klk6 expression levels of 32 normal and breast cancer tissues were detected and analyzed by the method and REST software.Results The amplification efficiencies for GAPDH and klk6 of the real-time quantitative PCR method were 0.90 And 0.95, respectively; inter-coefficient of variation were 1.0%~2.1% and 0.8%~1.2%,respectively; intra-coefficient of variation were 3.2% and 3.9%, respectively. The relative expression levels of klk6 in normal breast and breast cancer tissues were 0.017?0.009 and 0.040?0.017 with GAPDH as reference. Analysis results with REST indicated klk6 expression was up- regulated in breast cancer.Conclusion The real-time quantitative PCR method with Sybr Green I for klk6 mRNA quantification was simple, specific, reproductive and reliable, and could be used to study relationship betweeen klk6 expression and tumor.
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OBJECTIVE To discuss the best scheme for fungemia detection by analyzing and comparing the ability and performance characteristics of the BACTEC 9120 automated blood culture system and 4 kinds of blood culture bottles in the detection of simulated fungemia.METHODS Simulated blood culture was produced using 65 fungi isolates from clinical specimens and BACTEC Plus Aerobic/F,Plus Anaerobic/F,Peds Plus/F and Myco/F Lytic blood culture bottles and detected by BACTEC 9120 automated blood culture system.The final inoculum densities were 1-5 CFU/ml blood.The(time to detection TTD) of simulated blood culture with different concentrations of suspension produced using 2 kinds of standard strains and 4 kinds of blood culture bottles was compared.RESULTS From the 260 bottles in this study 216 had growth detected by the BACTEC 9120 blood culture system.The positive rates of BACTEC Plus Aerobic/F and Anaerobic/F,which were 90.77%and 41.54%,respectively,were significantly(P