Objective: To investigate the degree and distribution of tissue-resident CD8+ T cell (CD103+CD8+T cells) infiltration in colorectal cancer (CRC) tissues, and to analyze its relationship to patients’clinicopathological features and prognosis. Methods: Tissue chips of 88 cases of colon cancer tissues (No.HColA180Su14) and 77 cases of rectal cancer tissues (No. HRec-Ade180Sur-03) were obtained from Shanghai Outdo Biotech Co.,Ltd. Immunofluorescence staining was performed to examine the infiltration pattern and degree of CD103+CD8+T cells in the collected CRC tissues and their para-cancerous tissues. Wilcoxon rank test was used to compare CD103+CD8+T cell infiltration degree in CRC tissues and the para-cancerous tissues. Chi-square test was used to analyze the relationship between CD103+CD8+T cell infiltration in CRC and patients’clinicopathological features. Kaplan-Meier survival analysis was conducted to explore the correlation between CD103+CD8+T cell infiltration and patients’prognosis. Cox model was applied to analyze the correlation between different clinical parameters and patients’prognosis. Results: CD103+CD8+T cell infiltration presented no signifi ·cant differences between CRC and para-cancer tissues (P>0.05). Patients with distant metastasis had significantly lower CD103+CD8+T cell infiltration rate than patients without distant metastasis (P<0.01). There was no significant correlation between the infiltration of CD103+CD8+T cells and other clinicopathological features (P>0.05). Kaplan-Meier survival analysis showed that the overall survival (OS) of patients with high CD103+CD8+T cell infiltration was significantly longer than that of the patients with low infiltration (54.42% vs 25.00%, P<0.05). Multivariate Cox model analysis indicated that pathological grade (P<0.01) and high CD103+CD8+T cell infiltration (P<0.05) were independent prognostic factors for CRC. Conclusion： ：CD103+CD8+T cell infiltration in CRC is associated with patients’prognosis, suggesting that CD103+CD8+T cell plays an important role in the initiation and development of CRC.

调节性T（regulatory T，Treg）细胞是一类控制体内免疫反应性的T细胞亚群，在维持机体的免疫系统稳态和调节免疫 应答方面具有重要作用，并且发现在多种肿瘤类型中以较高比例存在，被认为是产生抗肿瘤免疫应答的主要障碍。Treg细胞在 其功能状态和稳定性方面存在异质性，通过多种机制发挥免疫负调控作用，目前在自身免疫和肿瘤免疫的研究中发现，特异性调 节不同Treg细胞群体可改善免疫疗效。但是，如何更加合理有效的以Treg细胞为靶点抑制肿瘤的进展仍需进一步探索。本文就 Treg细胞在肿瘤免疫中的作用机制及治疗应用新策略展开综述。

Objective: To investigate the degree of infiltration and distribution of tissue-resident CD8 + T cells (CD103 + CD8 + T cells) in gastric cancer tissues, and analyze the relationship between the degree of infiltration and clinicopathological features and prognosis. Methods: Tissue microarray and immunofluorescence staining were used to examine the CD8 + T cells and CD103 + CD8 + T cells infiltration in 90 cases of gastric cancer and their adjacent normal tissues. Wilcoxon rank test was used to compare the CD8 + T cells, CD103 + CD8 + T cells infiltration and CD103 + CD8 + T cells/CD8 + T cells ratio in gastric cancer and corresponding normal tissues. The chi-square test was used to analyze the relationship between CD8 + T cells, CD103 + CD8 + T cells infiltration and CD103 + CD8 + T cells/ CD8 + T cells ratio in gastric cancer tissues and clinicopathological features of the patients. Kaplan-Meier survival analysis was performed to explore the correlation between CD8 + T cells, CD103 + CD8 + T cells infiltration and CD103 + CD8 + T cells/ CD8 + T cells ratio and overall survival. Cox model was used to analyze the correlation between different clinical parameters and prognosis of the patients. Results: There was no significant difference for the infiltration of CD103 + CD8 + T cells between the gastric cancer tissues and adjacent normal tissues (P＞0.05). The infiltration rate of CD103 + CD8 + T cells in the cases in stage Ⅲ to Ⅳ (69.09%, 38/55) was significantly lower than that in the stage Ⅰ to Ⅱ cases (91.43%, 32/35), (χ 2 =6.175, P=0.013). There was no significant correlation between CD103 + CD8 + T cells infiltration and other clinicopathological features (P＞0.05). Kaplan-Meier survival analysis showed that the patients with high CD103 + CD8 + T cells infiltration showed significantly longer overall survival than the patients with low CD103 + CD8 + T cells infiltration (HR=2.187, 95%CI: 1.062-4.500, P=0.033 6). Multivariate Cox model analysis indicated that tumor diameter (HR=2.031, 95%CI: 1.163-3.546, P=0.013) and CD103 + CD8 + T cells infiltration (HR=0.516, 95%CI: 0.285-0.934, P=0.029) were independent prognostic factors for gastric cancer. Conclusion CD103 + CD8 + T cells in gastric cancer tissues should be associated with good prognosis, suggesting that they play an important role in the inhibition of gastric carcinogenesis and development, and can be used as an important factor for the prognosis evaluation of the patients with gastric cancer.

Objective: This study aimed to reveal the potential clinical and biological functions of frizzled related protein (FRZB) mRNA expression in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). Methods: We used the keyword “lung cancer” to search the data through Gene Expression Omnibus (GEO) database attached to NCBI(National Center of Biotechnology) and download the data of LUAD and LUSC from TCGA (The Cancer Genome Atlas) Database. A total of eight LUAD and six LUSC datasets were incorporated in this analysis. We defined cutoff value of FRZB using Cutoff Finder into the two groups to calculate hazard ratio (HR). Results: We found that high expression level of FRZB mRNA in tumor tissues was a positive prognostic factor for overall survival in LUAD [pooled HR（95%CI）=0.54（0.46-0.64）,P＜0.05 in univariate analysis; pooled HR（95%CI）=0.66（0.54-0.79）,P＜0.05 in multivariate analysis]. Interestingly, there was no similar results in LUSC [pooled HR（95%CI）=1.11（0.67-1.84）,P＞0.05 in univariate analysis; pooled HR（95%CI）=1.13（0.71-1.78）,P＞0.05 in multivariate analysis]. We also found that FRZB may inhibit WNT signal pathway by t-SNE and correlation analysis. By enrichment analysis, FRZB and its most correlated genes were involved in multiple immune-related pathways, such as complement and coagulation cascades, humoral immune response, etc. Conclusion High expression of FRZB mRNA in LUAD was associated with better prognosis of lung adenocarcinoma. These results suggest that FRZB may be used as a potential marker for favorable prognosis of lung adenocarcinoma.

IL-1有IL-1α和IL-1β两种亚型，其受体家族包括I型和II型两种受体形式（IL1R1和IL1R2）以及一种受体辅助蛋白 （IL1RAP）。在IL1RAP帮助下，IL-1α和IL-1β均可与ILR1形成IL-1/IL1R1/IL1RAP复合物，激活下游信号转导通路，发挥生物学 作用； IL1R2由于缺少胞内TIR结构域无法介导IL-1信号通路，而能通过与IL1R1竞争性结合抑制IL-1的作用。IL1R2起初被认 为是一种IL-1的诱捕受体，但在后续的研究中发现其在多种肿瘤中有异常表达，且多数呈现异常上调状态，仅在少数肿瘤中低表 达，其表达与许多肿瘤的发生发展以及预后有着重要关联。本文针对IL1R2在肿瘤发生发展中的作用方面的相关研究进展进行 综述。

Objective To purify the anti-T cell immunoglobulin mucin ( TIM)-3 monoclonal antibody 4E8 and examine its biological function in vitro. Methods The mouse monoclonal antibody against human TIM-3, clone 4E8, was obtained by standard protocol for monoclonal antibody purification. The cell lines expressing human TIM-3 molecule were obtained by cell transfection technique. We ex-amined the ability of 4E8 binding to human TIM-3 by flow cytometry. The ability of 4E8 blocking the binding of fusion protein TIM-3 Ig-huFc with phosphatidylserine( PtdSer) , the apoptotic cell surface TIM-3 ligand, was also analyzed by flow cytometry. Mixed lympho-cyte reaction ( MLR) and ELISA assays were used to determine the effect of TIM-3 monoclonal antibody ( 4E8) on IFN-γsecretion in CD4+ T cells. Results 4E8 specifically bound to human TIM-3 but could not block the binding of TIM-3 to Ptdser. Compared with the negative control (IFN-γ secretion: 958.3±153.2), 4E8 enhanced the ability of CD4+ T cells to secrete IFN-γ in MLR (4E8 of 10μg/mL group:IFN-γ secretion 2563±150.3 and 4E8 of 3.33 μg/mL group:IFN-γ secretion 1981±211.5) with statistically signifi-cant difference ( P<0.05) . In addition, the combined application of 4E8 with the anti-programmed death-1 ( PD-1) monoclonal anti-body nivolumab showed synergistic effects for increasing IFN-γ secretion in MLR assay ( 4E8 of 10 μg/mL group: IFN-γ secretion 3049±80.5 and 4E8 of 0.33μg/mL group:IFN-γsecretion 1957±321.3) as compared with 4E8 alone (10μg /mL group:IFN-γse-cretion 2563±150.3 and 0.33 μg/mL group:IFN-γ secretion 844±76.2) with statistically significant difference (P<0.05). Conclu-sion We successfully obtained a 4E8 clone of monoclonal antibody to human TIM-3 which may enhance the capacity of IFN-γsecre-tion from CD4+ T cells. The effect of enhancing IFN-γ secretion of CD4+T cells by TIM-3 monoclonal antibody was independent from blocking the binding of TIM-3 with Ptdser.

OBJECTIVETo explore the clinical efficacy and partial mechanism of medicinal-cake-separated moxibustion for senile osteoporosis.

METHODSSixty cases of senile osteoporosis were randomly divided into an observation group and a control group according to the random digits table, 30 cases in each one. The two groups were both treated with basic treatment of western medicine. The acupoints included four groups:① Dazhui (GV 14), Dazhu (BL 11) and Ganshu (BL 18); ② Zhongwan (CV 12), Danzhong (CV 17) and Zusanli (ST 36); ③ Pishu (BL 20), Shenshu (BL 23) and Mingmen (GV 4); ④ Shenque (CV 8) and Guanyuan (CV 4). Each group of acupoints was selected for one treatment. The observation group was treated with medicinal-cake-separated moxibustion, and the medicinal cake was consisted of fructus psoraleae (30 g), prepared rehmannia root (30 g), atractylodes (30 g), codonopsis pilosula (30 g), epimedium herb (20 g), rhizoma curculiginis (20 g), syzygium aromaticum (5 g) and cinnamon (5 g). The control group was treated with wheat-flour-cake moxibustion. Each acupoint was treated with 5 moxa cones in the two groups. The treatment was given once every other day for six months. The symptom score, lumbar and hip bone mineral density (BMD), serum type Ⅰ procollagen amino-terminal propeptide (PINP) and serum β-type Ⅰ collagen carboxy-terminal peptide (β-CTX) were observed before and after treatment.

RESULTSAfter treatment, the symptom score and serum β-CTX were significantly lowered (all<0.05), while the lumbar and hip BMD and serum PINP were significantly increased (all<0.05) of the two groups. After treatment, the symptom score and serum β-CTX in the observation group were significantly lower than those in the control group (both<0.05), while the lumbar and hip BMD and serum PINP in the observation group were significantly higher than those in the control group (all<0.05).

CONCLUSIONSThe medicinal-cake-separated moxibustion has significant efficacy for senile osteoporosis, which is superior to wheat-cake-se-parated moxibustion.

Gram-negative pathogen-induced nosocomial infections and resistance are a most serious menace to global public health. Qingfei Xiaoyan Wan (QF), a traditional Chinese medicine (TCM) formula, has been used clinically in China for the treatment of upper respiratory tract infections, acute or chronic bronchitis and pulmonary infection. In this study, the effects of QF on Pseudomonas aeruginosa-induced acute pneumonia in mice were evaluated. The mechanisms by which four typical anti-inflammatory ingredients from QF, arctigenin (ATG), cholic acid (CLA), chlorogenic acid (CGA) and sinapic acid (SPA), regulate anti-inflammatory signaling pathways and related targets were investigated using molecular biology and molecular docking techniques. The results showed that pretreatment with QF significantly inhibits the release of cytokines (TNF-α and IL-6) and chemokines (IL-8 and RANTES), reduces leukocytes recruitment into inflamed tissues and ameliorates pulmonary edema and necrosis. In addition, ATG was identified as the primary anti-inflammatory agent with action on the PI3K/AKT and Ras/MAPK pathways. CLA and CGA enhanced the actions of ATG and exhibited synergistic NF-κB inactivation effects possibly via the Ras/MAPK signaling pathway. Moreover, CLA is speculated to target FGFR and MEK firstly. Overall, QF regulated the PI3K/AKT and Ras/MAPK pathways to inhibit pathogenic bacterial infections effectively.

Background and objective Despite undergoing curative resection, the 5-year survival rate for stage III non-small cell lung cancer (NSCLC) patients is less than 25%. hTere is a need for biomarkers for prediction of survival and guiding individual therapy. MiR-155 is one of most commonly upregulated miRNAs in malignancies, and regulates multiple pro-oncogenic pathways. We aimed to investigate the prognostic impact of miR-155 in resected stage III NSCLC patients. Methods Tumor formalin-ifxed, paraffn-embedded (FFPE) from 162 resected stage III NSCLC patients were collected. Total RNA including miRNA was extracted, and qRT-PCR was used to determine the expression of miR-155. Results Spearman rank correlation test showed a positive correlation between miR-155 expression and nodal status (r=0.169, P=0.032). MiR-155 expression had a signiifcant prognostic impact in the total cohort (P<0.001), in squamous cell carcinomas (P=0.002) and in adenocarcinomas (P=0.003). In N0-1 subgroup, miR-155 expression did not have a signiifcant prognostic on overall survival in univariate analysis (P=0.319). In N2 subgroup, miR-155 had a negative prognostic effect on OS in univariate analysis (P<0.001). Cox regression analysis revealed that miR-155 expression was unfavorable prognostic factors of OS (RR=2.311, 95%CI:1.479-3.611, P<0.001). Conclusion High expression of miR-155 represents a valuable marker of poor clinical outcomes in patients with stage III NSCLC.

This study aims to clarify out the anti-inflammatory mechanism of Qingfei Xiaoyan Wan. Chemical constituents of Qingfei Xiaoyan Wan identified by UPLC Q-TOF, were submit to Molinspiration, PharmMapper and KEGG bioinformatics softwares for predicting their absorption parameters, target proteins and related pathways respectively; and the gene chip and real time-PCR were carried out to investigate the expression of inflammatory genes on lung tissue of guinea pigs or human bronchial epithelial cell lines. The predicted results showed that 19 of the 24 absorbable constituents affected at 9 inflammation-related pathways through 11 protein targets; Qingfei Xiaoyan Wan treatment can significantly reduce the infiltration of cytokines through ERK1 gene and 5 inflammatory pathways (Focal adhesion, Fc epsilon RI, Toll-like receptors, NK cell-mediated cytotoxic, and ERK/MAPK). The results of real time-PCR further confirmed that the anti-inflammatory effects of Qingfei Xiaoyan Wan were due to active ingredients such as arctigenin, cholic acid and sinapic acid intervened focal adhesion, Fc epsilon RI signaling and ERK/MAPK pathways. The novel approach of 'drug-target-pathway' will present an effective strategy for the study of traditional Chinese medicines.