Objective:To find the age-related indicators of human immune function in the blood, to explore the possibility to standardize the evaluation criterion of immune function in Chinese population.Methods:The peripheral blood samples from 478 healthy individuals with complete medical data collected at the Beijing Hospital from January 2019 to December 2019, were chosen and analyzed.The volunteers were divided into young(<40 years old)and elderly(≥70 years old)groups.The differences in RNA, cytokines and immune cell function(the proliferation of T cell as well as the ability of T cells and natural killer cells to lyse target cells)were compared between the young group versus the elderly group.Results:Real-time quantitative fluorescence RT-PCR detection method was used for detecting the following tumor immune-related genes in the peripheral blood of healthy people in young versus old groups: programmed death 1( PD1)gene, complement C3 gene( C3), complement C4 gene( C4), high sensitivity C-reactive protein gene( HsCRP), tumor necrosis factor alpha gene( TNFA), interferon gamma gene( INF- γ), interleukin-6 gene( IL-6), interleukin-8 gene( IL-8), cytotoxic T lymphocyte-associated antigen-4 gene( CTLA-4), nuclear transcription factor-κB gene( NF- κB), long intergenic noncoding RNA-erythroid pro-survival gene( lincRNA- EPS), long non-coding RNA-myeloid RNA regulator of Bim-induced death gene( lncRNA- Morrbid), lincRNA-cyclooxygenase 2 gene( lincRNA- Cox2), Lnc-Dendritic cells( Lnc- DC), and circRNA-7 gene( ciRS-7). The mRNA level of C4 was significantly lower in elderly people than in young controls[young group(1.01±0.18) vs.old group(0.64±0.13), P=0.047], while others showing a non-statistically significant difference( P>0.05). The protein levels of programmed death 1(PD1), complement C3(C3), complement C4(C4), high sensitivity C-reactive protein(HsCRP), tumor necrosis factor alpha(TNF-α), interferon gamma(INF-γ), interleukin-6(IL-6), interleukin-8(IL-8), cytotoxic T lymphocyte-associated antigen-4(CTLA-4)and nuclear transcription factor-κB(NF-κB)in plasma were detected by enzyme-linked immunosorbent assay(ELISA). The following protein levels were significantly higher in the elderly group than in the young group: IL-6 protein[young group(249.30±100.02)ng/L vs.old group(283.00±94.98)ng/L, P=0.021], HsCRP protein[young group(2543.00±1111.89)ng/L vs.old group(3056.00±1056.61)ng/L, P=0.002], INF-γ protein[young group(362.40±383.67)ng/L vs.old group(500.40±502.27)ng/L, P=0.047]and TNF-α protein[young group(20.74±29.47)ng/L vs.old group(33.09±48.91)ng/L, P=0.042]. While the level of C4 was significantly lower in the elderly group than in the young group[young group(449.50±51.17)ng/L vs.old group(407.10±59.78)ng/L, P<0.0001]. T-cell proliferation, quantified by flow cytometry-based fluorescent dye dilution, showed that the CD3 + T cells of elder people had proliferated through 4 generations, while the CD3 + T cells of young people had proliferated through 5 generations.As shown in the NK killing assays, the secretion of LDH in the target A549 tumor cells decreased significantly after treated with NK cells belonging to elder individuals.The result demonstrated that individuals over 70 years old have significantly lower levels of the killing activity of NK cells( P<0.05). However, there was not statistically significant in T lymphocyte killing activity between different ages( P>0.05). Conclusions:IL-6, HsCRP, INF-γ and TNF-α levels are increased with aging, while the level of C4 significantly is decreased.The proliferation ability of T cells and the ability of NK cells to kill tumor cells are weakened.

[摘 要] 目的：基于已发表的芯片数据通过生物信息学方法筛选差异表达基因，以发现前列腺癌诊断/预后和耐药相关分子标志物。方法：筛选GEO数据库中已发表的前列腺癌mRNA芯片数据GSE6956和前列腺癌细胞多烯紫杉醇耐药mRNA芯片数据GSE33455进行差异表达分析；通过生物学功能注释、基因通路富集分析、蛋白质相互作用网络（protein-protein interaction，PPI）分析等生物信息学方法发现和识别与差异表达基因相关的生物学功能和信号通路；比对TCGA数据库，验证差异表达基因在前列腺癌组织及癌旁组织中的表达，并通过Kaplan-Meier分析差异表达基因对前列腺癌患者生存率的影响；用qPCR方法验证差异表达基因在前列腺癌细胞株PC3及多烯紫杉醇耐药细胞PC3-DTX中的表达情况。结果：共筛选出227个在前列腺癌和前列腺癌多烯紫杉醇耐药细胞芯片数据中共同差异表达基因。差异表达基因主要富集到了癌症相关通路（Lysosome、Sphingolipid、FoxO、Acute myeloid leukemia），并主要参与细胞黏附、自噬和胞内蛋白转运等生物学过程。构建PPI网络选取18个连接度最高的基因作为Hub基因。Hub基因和共同差异表达基因中，上调基因CITED2、LRP12和RPL17-C18orf32与前列腺癌患者的不良预后显著相关。qPCR验证显示CITED2在多烯紫杉醇耐药细胞PC3-DTX中高表达。结论：通过生物信息学方法筛选出在前列腺癌组织和耐药细胞中共同差异表达，且与前列腺癌患者的不良预后密切相关的基因，为前列腺癌诊断/预后和耐药分子标志物的研究提供了新的思路。

PURPOSE: The significance of neuroendocrine differentiation (NED) in gastric carcinoma (GC) is controversial, leading to ambiguous concepts in traditional classifications. This study aimed to determine the prognostic threshold of meaningful NED in GC and clarify its unclear features in existing classifications. MATERIALS AND METHODS: Immunohistochemical staining for synaptophysin, chromogranin A, and neural cell adhesion molecule was performed for 945 GC specimens. Survival analysis was performed using the log-rank test and univariate/multivariate models with percentages of NED (PNED) and demographic and clinicopathological parameters. RESULTS: In total, 275 (29.1%) cases were immunoreactive to at least 1 neuroendocrine (NE) marker. GC-NED was more common in the upper third of the stomach. PNED, and Borrmann's classification and tumor, lymph node, metastasis stages were independent prognostic factors. The cutoff PNED was 10%, beyond which patients had significantly worse outcomes, although the risk did not increase with higher PNED. Tumors with ≥10% NED tended to manifest as Borrmann type III lesion with mixed/diffuse morphology and poorer histological differentiation; the NE components in this population mainly grew in insulae/nests, which differed from the predominant growth pattern (glandular/acinar) in GC with <10% NED. CONCLUSIONS: GC with ≥10% NED should be classified as a distinct subtype because of its worse prognosis, and more attention should be paid to the necessity of additional therapeutics for NE components.
Adenocarcinoma ; Chromogranin A ; Classification ; Humans ; Immunohistochemistry ; Lymph Nodes ; Neoplasm Metastasis ; Neural Cell Adhesion Molecules ; Prognosis ; Stomach ; Stomach Neoplasms ; Synaptophysin

Objective To explore the application of carbon nanoparticles for tracing sentinel node(SN) in early gastric cancer (EGC).Methods Forty-six patients with EGC conducted subserous injection of carbon nanoparticle solution 1.0 mL at 4-6 points around tumor during operation.SN was examined by adopting 3 different methods.The first stained lymph node was defined as the first SN(FSN),and the whole group lymph nodes were identified as the group SN(GSN) and all lymph nodes colored within 5 min were labeled as the all SN(ASN).The affirmative SN and other resected lymph nodes were performed the histopathological examination and comparison.Resnlts In all 46 cases,SN was successfully detected out with the detection rate of 100％;the number of SN,GSN and ASN were 1.04 ± 0.21,3.93-±-1.90 and 13.61 ± 4.36 respectively,the difference in SN number among 3 different methods had statistical significance(P＜0.05).The accuracy rates of FSN,GSN and ASN for judging perigastric lymph node metastasis status were 95.65％,100％ and 100％,respectively.Conclusion The carbon nanoparticles tracing SN in EGC has higher staining rate and higher accuracy for predicting lymph node metastasis;GSN has high accuracy,moreover has less trauma and is most suitable method for detecting SN.

Currently, clinical laboratories are developing towards two different directions .One is centralized laboratory with automated pipelines , and the other is highly integrated portable detection system.The latter evolved from an early single test device gradually to an integrated platform based on microfluidic technology , which can operate multiple steps and tests simultaneously .The microfluidic system is further developed to a miniaturized device which can perform biochemical , immune, microbial and cytological tests.With the medical reform process in China , the microfluidic systems will appear in the primary health unit or even families .This review concludes the current application of microfluidic technology in clinical diagnostics and its future trends .

Objective In order to establish a basis for exploring diagnostic biomarkers of type 2 diabetes mellitus (T2DM), our research screened differentially expressed circRNAs in high glucose treated human umbilical vein endothelial cells ( HUVECs) and validated these circRNAs in both cell models and peripheral blood samples of T 2DM patients.Methods The research used HUVECs as experimental model . The control group (n=3) and high glucose group (n=3) were set up and treated under normal condition (5.5 mmol/L) and high glucose condition (30.0 mmol/L), respectively.The research utilized high-throughput sequencing technology to preliminarily screen differentially expressed circRNAs .Differentially expressed circular RNAs were validated in the endothelial cell model by PCR and real -time quantitative PCR ( q-PCR) techniques.Subsequently , in the peripheral blood samples of T 2DM patients ( n=32) and control individuals ( n=28 ), the differentially expressed circRNAs were further validated . Finally, t-Test, correlation analysis and ROC curve were used to analyze the experimental results .Results Total 1087 differentially expressed circular RNAs were screened by high-throughput sequencing;and of these , 554 were up-regulated, 533 were down-regulated.Six increased circRNAs were selected and validated in HUVEC model;and their PCR and q-PCR results matched with sequencing results .Further validation was conducted in peripheral blood samples for three most up-regulated circRNAs.This study found that the differential expression of hsa_circ_0031739 in peripheral blood samples was statistically significant ( 0.015 ±0.0025 vs.0.006 ±0.0013,P=0.0059 <0.05).The area under the ROC curve (AUC) was 0.730 and the expression level of hsa_circ_0031739 was positively correlated with blood glucose ( GLU ) and glycated hemoglobin ( GHb ) ( r=0.317, P=0.0137 <0.05;r=0.348, P=0.0064 <0.05 ) .Conclusion HUVECs were commonly utilized as cell models in the study of T 2DM, and differentially expressed circular RNA profiles existed afterhigh glucose treatment .The differential expression of hsa_circ_0031739 was significant in both high glucose treated HUVECs and peripheral blood samples of T 2DM patients, and the expression level of hsa_circ_0031739 was correlated with GLU and GHb , which has certain diagnostic significance.Therefore, hsa_circ_0031739 may become a new diagnostic biomarker for T2DM and be helpful for the comprehensive diagnosis and mechanism research of T 2DM.

Objectives To analyze the relationship between phosphorylation of serine-3 of cofilin1 and Taxol resistance of ovarian cancer and to evaluate the prognostic value of cofilin1 phosphorylation in estimating ovarian cancer survival using clinical samples.Methods Wild type (WT) plasmids with over-expression of wild cofilin1,S3L plasmids with over-expression of cofilin1 and inhibited phosphorylation at serine-3,and siRNA-C plasmids with down-regulation of cofilin1 were constructed using molecular biology techniques.After the SKOV3 and SK-TR30 cell-lines were transfected with these plasmids,changes in biological characteristics and drug resistance after instant transfection were compared.Fifty-one elderly female patients with microscopically confirmed ovarian cancer,who had received chemotherapy with Taxol and cis-platinum (TC)after surgery,including 30 chemo-sensitive and 21 chemo-resistant cases,were recruited in this study.Immunohistochemical methods were used to detect the expression of cofilinl and phosphorylated cofilin in tissue sections from the two groups.Progression free survival(PFS)was analyzed.Results Compared with the control group,the proportion of cells increased at the G0/G1 phase(P=0.034)and decreased at the S phase (P=0.031),and the apoptosis rate decreased(P=0.020),in SKOV3 cells with high expression of cofilin1.However,these characteristics disappeared when the wild type was replaced with the inactive mutant S3L.When cofilin1 expression was down-regulated in SK-TR30 cells,the proportion of cells at the G0/G1 phase decreased(P=0.020).The expression of cofilin1 was detected in 56.7 ％ (17/30)and 57.1％ (12/21),respectively,sections of ovarian tumor tissues of the chemo-sensitive and chemoresistant groups,and there was no statistic difference (x2=0.332,P =0.943)between the two groups.The expression of phosphorylated cofilin was much higher in the chemo-resistant group (85.7％ or 18/21)than in the chemo-sensitive group (53.3％ or 16/30),and the difference was statistically significant(x2=5.829,P=0.016).The higher expression of phosphorylated cofilin was also correlated with shorter PFS(x2 =21.440,P＜0.01,95％ CI:0.068-0.883),especially in the chemo-sensitive group.Conclusions Serine-3 phosphorylation status of cofilin 1 is associated with paclitaxel resistance in ovarian cancer,but the underlying mechanisms of regulation need further investigation.

OBJECTIVE:To investigate the effects of Fluorouracil implants on intraperitoneal chemotherapy safety of advanced gastric cancer patients. METHODS:The information of 264 patients with advanced gastric cancer were analyzed retrospectively. All patients were given Fluorouracil implants 800 mg,scattered into the mesenteric and surgical area,dose of each implantation site less than 200 mg,the distance of implantation range from incision and anastomosis more than 3 cm. All patients were given second generation cephalosporin 30 min before operation,at 3rd h during operation,1-3 d after operation. The occurrence of complications and ADR during hospitalization were observed after operation. RESULTS:After operation,6 patients suffered from complications, with incidence of 2.28%,including 3 cases of gastrointestinal reaction,2 cases of anastomotic bleeding and 1 case of anastomotic leakage. Routine blood test and liver and kidney function were rechecked 3 d after operation,mainly thrombocytopenia and liver function injury,with the incidence of 2.65%. CONCLUSIONS:Fluorouracil implants can be used for intraperitoneal chemotherapy of advanced gastric cancer patients. However,attention should be paid to thrombocytopenia and liver function injury.

Objective To study the clinical management way for HPV+/papanicolaou (Pap)-during cervical cancer screening.Methods To analyze retrospectively the data from the patients who had loop electrical excision procedure (LEEP) for biopsy confirmed cervical intraepithelial neoplasia (CIN)Ⅱ in Peking University People's Hospital from Jan.2010 to Dec.2014.Results (1) For biopsy confirmed CINⅡ,HPV positive rate was 98.5％ (135/137),Pap test positive [≥atypical squamous cell of undetermined significance (ASCUS)] rate was 69.3％ (95/137),there was significant difference between them (x5=43.32,P＜0.01).(2) For the 42 patients with HPV+/Pap-,whose cytology slides were reviewed again.Among them,the interpretations of there were 16 cases confirmed as the same before,while 26 cases were changed to abnormal (≥ASCUS).Cytology be misdiagnosed was 19.0％ (26/137) at the first review.Among the 26 cases,13 (50.0％) cases were missed for the little amount of abnormal cells,8 (30.8％) cases for mild atypical morphology changed;the other 5 (19.2％) cases missed for stain problems.(3) For the cervical LEEP samples,37 cases of the pathology diagnosis were upgrade to CIN Ⅲ+,among them,2 cases of microinvasive cervical carcinoma,1 case of invasive cancer,34 cases of CIN Ⅲ;37 cases were CINⅠ or no lesion found;63 cases were still CIN Ⅱ.Four to six months later after LEEP,the cytology abnormal rate was 11.7％ (16/137),and the HR-HPV positive rate was 34.3％ (47/137).Conclusions Compared with cytology alone,cytology combined with HPV testing increase the sensitivity of cervical high grade lesion.For the cases of HPV+/Pap-cases,the cytology slides should be reviewed.The quality control of cervical exfoliate sample collection and interpretation should be strengthened.LEEP procedure is not only a treatment method,but also it could provide samples to confirm the diagnosis.

Objective To investigate the characteristics of the diagnosis and treatment center for Ebola observing ward and reduce infection risk according to allocation scheduling method of nursing staff. To provide reference for other similar tasks. Methods We used the method of grouping fixed group scheduling method and fixed working hours to identify and refine the content of nursing work. Results A total of 65 cases of suspected Ebola including 5 cases diagnosed as EVD patients were cured. No case of infection occurred in nursing staff. Conclusions During the infectious disease nursing work, reasonable personnel allocation and scheduling method can make the nursing staff work with full energy and strength and ensure the realization ofzero infectiontargets.