Objective:To observe the baseline characteristics and clinical efficacy in cancer patients who were treated with immune checkpoint inhibitors (ICIs) and experienced immune-related adverse events (irAEs).Methods:The clinical efficacy in cancer patients experiencing irAEs was retrospectively analyzed, and the objective response rate, progression-free survival, and overall survival were evaluated.Results:The median onset time of irAEs in 23 patients was 3.17 months. There were 47.8% (11/23) patients with multi-system irAEs, 63.3% (7/11) of which were non-simultaneous. The most common irAEs were immune-related pneumonia, hypothyroidism and immune-related liver dysfunction. Complete remission, partial remission and stable disease were respectively achieved in 0, 10 and 11 cases, while the other two patients developed progressive disease. The objective response rate was 43.5% and the disease control rate was 91.3%. With a median follow-up period of 16.5 months, 17 patients (73.9%) had progressive disease and the median progression-free survival was 9.63 months. Eight patients (34.8%) died before reaching the median overall survival. The progression-free survival and overall survival of patients with irAEs ≥grade 3 were significantly shorter than those with irAEs of grade 1-2 ( P<0.01). Conclusions:The occurrence of irAEs might correlate with the short-term efficacy and clinical benefits in patients treated with ICIs.

Objective To detect exon deletions/duplications in the DMD gene in Duchenne and Becker muscular dystrophy pedigrees using multiplex ligation-dependent probe amplification method,and explore the usefulness of multiplex ligation-dependent probe amplification analysis as a method for genetic diagnostics in patients with Duchenne and Becker muscular dystrophy.Methods Exon deletions/duplications in the DMD gene were analyzed by multiplex ligation-dependent probe amplification for two pedigrees with Duchenne muscular dystrophy and Becker muscular dystrophy.Patients and carriers were diagnosed by multiplex ligation-dependent probe amplification.Results The pedigree with Duchenne muscular dystrophy was caused by DelEx45-50 mutation,while the pedigree with Becker muscular dystrophy was caused by Dup Ex3-4 mutation.Patients and carriers were diagnosed by multiplex ligation-dependent probe amplification method.Conclusion Exon deletions/duplications in the DMD gene can be indicated by probe copies using multiplex ligation-dependent probe amplification method under standard operating procedure.Multiplex ligation-dependent probe amplification should be considered as a rapid and accurate clinical method for an initial mutation analysis of DMD gene with exon deletions/duplications.

Objective To develop a new method to detect A (TA)n TAA polymorphism in the UGT1A1 gene promoter by fluo‐rescence real‐time quantitative PCR (RQ‐PCR) .Methods Genomic DNA was extracted from peripheral blood in 16 patients with Gilbert′s syndrome and 66 healthy individuals .The polymorphic A(TA)n TAA sequence in the UGT1A1 gene promoter was deter‐mined by DNA sequencing .A pair of primers and two TaqMan probes labeled with either 5′FAM or VIC reporter dye incorporated a 3′minor groove binder were designed .The A(TA)n TAA polymorphisms in the UGT1A1 gene promoter were identified by RQ‐PCR for all research subjects .The sensitivity and specificity of RQ‐PCR for detecting the A(TA)nTAA polymorphisms were veri‐fied by DNA sequencing method .Results The homozygous A(TA)7TAA polymorphism was found in the promoter region of the UGT1A1 gene in all 16 patients with Gilbert′s syndrome by using RQ‐PCR .The homozygous A(TA)6TAA polymorphism was foundin46healthysubjects,whiletheheterozygousA(TA)6TAA/A(TA)7TAApolymorphismwasfoundinother20healthysub‐jects .All A(TA)nTAA polymorphisms in the promoter region of the UGT1A1 gene identified by RQ‐PCR were consistent with that of DNA sequencing .Conclusion It is a sensitive ,specific and simple method to detect the A (TA)n TAA polymorphisms in the promoter region of the UGT1A1 gene by RQ‐PCR ,which can be promoted and applied in clinic .

Objective To investigate the correlation between MicroRNA-146a (miR-146a) C ＞ G polymorphism and ischemic stroke.Methods The case control studies of the relationship between miR-146a polymorphism and ischemic stroke published before February 2016 were retrieved comprehensively.The Statal2.0 software package was used to conduct the meta-analysis.The odds ratio (OR) and 95％ confidence interval (CI) were used to evaluate the strength of association between the polymorphisms and the risk of ischemic stroke.Results A total of 8 articles were enrolled,including 2 891 patients in the case group and 4 019 in the control group.The selected literature did not have obvious publication bias.In the general population,the dominant model (GG + CG vs.CC:OR 1.011,95％ CI 0.863-1.185;P =0.889),recessive model (GG vs.CG + CC:OR 0.999,95％ CI 0.761-1.311;P=0.994),heterozygous model (CG vs.CC:OR 1.052,95％ CI 0.943-1.173;P =0.368),homozygous model (GG vs.CC:OR 1.114,95％ CI 0.819-1.515;P =0.491),and allele model (G vs.C:OR 1.062,95％ CI0.919-1.227;P=0.413) did not show significant correlation between the miR-146a C ＞ G polymorphism and the risk of ischemic stroke.Subgroup analysis showed that the miR-146a C ＞ G polymorphism was not associated with the onset risks of large artery atherosclerotic and small arterial occlusive stroke.Conclusions According to the literature available,the miR-146a C ＞ G polymorphism may not be significantly associated with the risk of ischemic stroke.

Objective To investigate the correlation between methylenetetrahydrofolate reductase (MTHFR) gene 3 '-untranslated region rs4846049 G/T polymorphism and risk of ischemic stroke in a Chinese Han population. Methods A total of 396 patients with ischemic stroke and 378 healthy subjects (control group ) were selected using a case-control study design. Large artery atherosclerosis and small artery occlusion in the case group were 268 and 128 cases, respectively. Polymerase chain reaction-restriction fragment length polymorphism and the direct sequencing method were used to detect MTHFR gene rs4846049 G/T polymorphism. Results As compared to the GG genotype, the TT genotype significantly increased the risk of ischemic stroke (odds ratio [OR] 2. 87, 95% confidence interval [CI] 1. 43-5. 76;P=0. 003). Compared with G allele, T allele significantly increased the risk of the disease (OR 1. 62, 95% OR 1. 28-2. 06; P< 0. 001 ). Subgroup analyses showed that the rs4846049 G/T polymorphism could significantly increase the onset risks of LAA and SAO subtype stroke (all P<0.05). Conclusions MTHFR gene rs4846049 G/T polymorphism may be associated with the increased susceptibility to ischemic stroke in the Chinese Han population. The T allele may be a genetic risk factor for ischemic stroke in the Chinese Han population.

Objective To detect the expressions of circulating microRNAs (miRNAs) of peripheral whole blood in Alzheimer's disease (AD) and investigate the potential roles of the miRNAs as diagnostic bi omarkers for Alzheimer's disease.Methods Peripheral blood samples were obtained from 110 AD patients and 150 age-and gender-matched normal controls.The concentrations of seven candidate miRNAs,including miR-9,miR-29a,miR-29b,miR-101,miR-181c,miR-137and miR-126,were measured with a real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) method.The data of two groups were collected and analyzed by SPSS 19.0 software.Results It was found that miR-29a (P=1.12×10-5) and miR-101 (P=6.24× 10-7) were markedly down-regulated in peripheral blood of AD patients compared with normal controls.In addition,logistic regression analysis revealed peripheral whole blood miR-29a/101 combination could be a potential biomarker of AD with better specificity (82％) and sensitivity (75％).Conclusions miR-29a/101 combination in peripheral whole blood may serve as a useful noninvasive diagnostic biomarker for AD.

BACKGROUNDAltered immunoresponse is associated with tumorigenesis and cancer progression. This study assessed the levels of tumor-infiltrating CD3 + or CD8 + T lymphocytes and interleukin-2 (IL-2) protein in radically resected non-small cell lung cancer (NSCLC) tissues to predict overall survival (OS) of the patients.

METHODSParaffin-embedded tissue specimens from 129 NSCLC patients were retrospectively collected for immunostaining of CD8 + , CD3 + , and IL-2 expression. Clinicopathological and survival data were collected and analyzed using the Chi-squared test, Kaplan-Meier curves, and the log-rank test or the Cox regression model.

RESULTSThe data showed a significant inverse association between CD8 + T lymphocyte levels and IL-2 expression (r = -0.927; P = 0.000) and between the levels of CD8 + and CD3 + T lymphocytes (r = -0.722; P = 0.000), but a positive association between CD3 + T lymphocyte levels and IL-2 expression (r = 0.781; P = 0.000) in NSCLC tissues. Furthermore, the levels of CD3 + and CD8 + T lymphocytes and IL-2 expression were associated with tumor stage (P = 0.023, 0.006, and 0.031, respectively) and the level of CD8 + T lymphocytes was associated with the patient gender (P = 0.024). In addition, the levels of CD8 + T lymphocytes were associated with an unfavorable 5-year OS, whereas patients with high levels of CD3 + T lymphocytes in tumor lesions and IL-2-expressing tumors had significantly better 5-year OS rates than patients with low levels.

CONCLUSIONSThe levels of CD8 + T cells in tumor lesions and IL-2 expression were both independent predictors of OS for these NSCLC patients. Thus, the detection of tumor-infiltrating CD3 + or CD8 + T lymphocytes and IL-2 expression could be useful to predict the prognosis of radically resected NSCLC patients.

CD3 Complex ; metabolism ; CD8-Positive T-Lymphocytes ; metabolism ; Female ; Humans ; Immunohistochemistry ; Interleukin-2 ; metabolism ; Lung Neoplasms ; immunology ; metabolism ; Lymphocytes, Tumor-Infiltrating ; metabolism ; Male ; Prognosis

Objective To investigate the correlation between R219K (rs2230806 G/A) polymorphism in the ATP binding cassette transporter (ABC) A1 gene and ischemic stroke in a Chinese Han population. Methods A total of 360 patients with ischemic stroke and 358 healthy subjects were selected using a case-control study design. The patients with ischemic stroke were redivided into either a large artery atherosclerosis (LAA) group or a smal artery occlusion (SAO) group according to the TOAST criteria. Polymerase chain reaction-restriction fragment length polymorphism analysis and direct sequencing method were used to detect R219K (rs2230806 G/A) polymorphism in the ABCA1 gene. Results Using GG genotype as a reference, the AA genotype reduced the risk of ischemic stroke by 65% (odds ratio [OR] 0. 35, 95%confidence interval [CI] 0. 23 - 0. 55; P < 0. 001) and the onset risk of LAA by 77% (OR 0. 23, 95% CI 0. 13 - 0. 40; P < 0. 001), but it did not have significant correlation with SAO. The interaction of rs2230806 G/A polymorphisms with smoking, drinking, hypertension, and diabetes had no significant effect on the on-set risk of ischemic stroke (al P > 0. 05). AA genotype was enable to increase the high-density lipoprotein cholesterol levels of the patient group (OR 0. 35, 95% CI 0. 28 - 0. 42; P < 0. 001) and the control group (OR 0. 19, 95% CI 0. 14 - 0. 23; P < 0. 001) significantly, while it did not have significant correlation with the low-density lipoprotein cholesterol, total cholesterol, and triglyceride. Conclusions R219K (rs2230806 G/A) polymorphism in the ABCA1 gene may be associated with the reduced predisposition of ischemic stroke in a Chinese Han population, especialy LAA. The A alele may be a hereditary protective factor; its mechanism may be associated with the increase of high-density lipoprotein cholesterol levels.

OBJECTIVETo evaluate the clinical value of T lymphocyte subsets in prediction of chemotherapy responses of patients with pulmonary adenocarcinoma.

METHODSFifty-five chemotherapy-naive patients with pathologically or cytologically confirmed pulmonary adenocarcinoma were examined for peripheral blood T lymphocyte subsets using flow cytometry, including CD3(+) T cells, CD3(+)CD4(+) T cells, CD3(+)CD8(+) T cells, CD45RO(+) T cells and CD45RA(+) T cells.

RESULTSPatients who responded favorably to chemotherapy (CR(+)PR) showed a significantly lower percentage of CD45RA(+) T cells than those who failed to respond to chemotherapy (P=0.04). CD45RO(+) T cell percentage were slightly higher in the response group than in the non-response group, but this difference was not statistically significant (P=0.25). The other T cell subsets, namely CD3(+), CD3(+)CD4(+), and CD3(+)CD8(+) T cells showed no significant differences between the two groups.

CONCLUSIONA high percentage of peripheral blood CD45RA(+) T cells is associated with a poor short-term outcome of chemotherapy in patients with advanced pulmonary adenocarcinoma. Peripheral blood CD45RA(+) T cell level can be a reliable index for predicting chemotherapy efficacy in these patients.

Adenocarcinoma ; blood ; immunology ; Adult ; Aged ; Female ; Humans ; Leukocyte Common Antigens ; metabolism ; Lung Neoplasms ; blood ; immunology ; Lymphocyte Count ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology

Objective To investigate the effect of cerebral ischemic preconditioning (IP) on the expressions of angiopoietin-1 (Ang-1) and its receptor Tie-2 mRNA in cerebral ischemia in rats.Methods Ninety-nine Wistar rats were randomly assigned to three groups:sham operation (n =9),non-ischemic preconditioning (NIP) (n =45),and IP (n =45).The latter two groups were redivided into 5 subgroups:ischemia-reperfusion 1,3,7,14,and 21 days (n =9 in each group).A model of transient middle cerebral artery occlusion (MCAO) was induced by the intraluminal suture method for focal IP (ischemia for 10 minutes and restoring perfusion).Infarct volume was determined by 2,3,5-triphenyltetrazolium staining.The expression levels of Ang-1/Tie-2 mRNA were detected by in situ hybridization.Results The infarct volumes in the 1 -,3-,and 7-day subgroups of the IP group were significantly smaller than those in the relative subgroups of the NIP group (all P＜ 0.05).The expression of Ang-1 mRNA in the 3- and 7-day subgroups of the IP group and the expression of Tie-2 mRNA in the 1-,3-,and 7-day subgroups of the NIP group were upregulated significantly (all P ＜ 0.05).The infarct volume in the 3-day subgroup of the IP group was reduced most significantly (P ＜ 0.05).The expression of Ang-1 mRNA in the 7-day subgroup was upregulated significantly,and the peak expression of its receptor Tie-2 mRNA appeared at day 3 after IP and continued to day 7.Pearson correlation analysis showed that the expression levels of Ang-1/Tie-2 mRNA were significantly negatively correlated with infarct volume (P ＜0.01).Conclusions The expression of Ang-1/Tie-2 mRNA in the IP group was upregulated within the time window of ischemic tolerance (1 - 7 days after preconditioning),in which Ang-1 may mainly act on the later stage of the cerebral ischemic tolerance.