Objective To investigate the expression of Fascin in early-stage NSCLC, evaluate the relevance between Fascin expression level and prognosis.Methods The immunohistochemistry method was used to assess the expression of Fas-cin in 111 lung cancer FFPE tissues with stage Ⅰ and Ⅱ NSCLC.The relationship between Fascin expression and the clinico-pathological characteristics was analyzed.The prognostic significance of Fascin expression was evaluated with Kaplan-Meier sur-vival analysis.Results In the early-stage of NSCLC, the positive rate of Fascin expression was 64.8%, no expression in the paracarcinoma tissue.The positive rate of squamous cell carcinoma was 78.7% and was significantly higher than that in adeno-carcinoma 48.0%(P<0.01).Whether in squamous cell carcinoma or adenocarcinoma group, the expression of Fascin was correlated significantly with lymph node metastasis tumor stages and DFS(P<0.05).And the positive expression of Fascin was an independent risk factor of poor prognosis for patient with NSCLC .Conclusion Fascin is expected to be a biomarker for the prognosis of patients with early-stage NSCLC.

OBJECTIVEThe aim of this study was to detect the plasma concentration of OLC1 (overexpressed in lung cancer 1) protein as a potential cancer biomarker, and evaluating its clinical application value in the diagnosis of non-small cell lung cancer (NSCLC).

METHODSWe prepared OLC1 antibody with OLC1 full length protein, in 5-6-week old Bal B/c mice. Each mouse was immunized four times at a dose of 15-30 µg antigen protein, and the interval between two consecutive immunizations was two weeks. Antibody screening was made by ELISA and Western blot, and a double antibody sandwich ELISA kit was developed. We used this established ELISA kit to detect the plasma concentration of OLC1 protein in 281 NSCLC patients and 92 gender- and age-matched healthy controls. Area under the receiver operating characteristic curve (AUC) was used to evaluate the detection efficacy of OLC1.

RESULTSWe obtained 11 OLC1 monoclonal antibodies and successfully established the ELISA kit to detect the plasma concentration of OLC1 with a detection range from 1.95 ng/ml to 62.50 ng/ml. OLC1 concentration in the case group (124.69 ng/ml) was significantly higher than that in the control group (67.07 ng/ml, P < 0.001). In the scenario of distinguishing NSCLC from control group, AUC result was 0.69. When the cut-off was set at 67.72 ng/ml, the sensitivity and specificity was 84.4% and 51.1%, respectively. In term of distinguishing early lung cancer (IA) from normal controls, the AUC, sensitivity and specificity were 0.68, 77.8% and 54.4%, respectively.

CONCLUSIONThe plasma concentration of OLC1 protein is significantly elevated in NSCLC patients. OLC1 may be as a potential cancer biomarker applied in clinical diagnosis.

Adult ; Animals ; Antibodies, Monoclonal ; Biomarkers, Tumor ; blood ; Blotting, Western ; Carcinoma, Non-Small-Cell Lung ; blood ; diagnosis ; immunology ; Early Detection of Cancer ; methods ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Lung Neoplasms ; blood ; diagnosis ; immunology ; Male ; Mice, Inbred BALB C ; Middle Aged ; Oncogene Proteins ; blood ; immunology ; ROC Curve ; Sensitivity and Specificity ; Young Adult

Objective The aim of this study was to detect the plasma concentration of OLC 1 ( overexpressed in lung cancer 1 ) protein as a potential cancer biomarker , and evaluating its clinical application value in the diagnosis of non-small cell lung cancer (NSCLC).Methods We prepared OLC1 antibody with OLC1 full length protein, in 5-6-week old Bal B/c mice.Each mouse was immunized four times at a dose of 15-30μg antigen protein , and the interval between two consecutive immunizations was two weeks.Antibody screening was made by ELISA and Western blot , and a double antibody sandwich ELISA kit was developed .We used this established ELISA kit to detect the plasma concentration of OLC 1 protein in 281 NSCLC patients and 92 gender-and age-matched healthy controls .Area under the receiver operating characteristic curve (AUC) was used to evaluate the detection efficacy of OLC 1.Results We obtained 11 OLC1 monoclonal antibodies and successfully established the ELISA kit to detect the plasma concentration of OLC1 with a detection range from 1.95 ng/ml to 62.50 ng/ml.OLC1 concentration in the case group (124.69 ng/ml) was significantly higher than that in the control group (67.07 ng/ml, P<0.001).In the scenario of distinguishing NSCLC from control group , AUC result was 0.69.When the cut-off was set at 67.72 ng/ml, the sensitivity and specificity was 84.4%and 51.1%, respectively.In term of distinguishing early lung cancer (IA) from normal controls, the AUC, sensitivity and specificity were 0.68, 77.8% and 54.4%, respectively .Conclusion The plasma concentration of OLC 1 protein is significantly elevated in NSCLC patients.OLC1 may be as a potential cancer biomarker applied in clinical diagnosis .

Objective The aim of this study was to detect the plasma concentration of OLC 1 ( overexpressed in lung cancer 1 ) protein as a potential cancer biomarker , and evaluating its clinical application value in the diagnosis of non-small cell lung cancer (NSCLC).Methods We prepared OLC1 antibody with OLC1 full length protein, in 5-6-week old Bal B/c mice.Each mouse was immunized four times at a dose of 15-30μg antigen protein , and the interval between two consecutive immunizations was two weeks.Antibody screening was made by ELISA and Western blot , and a double antibody sandwich ELISA kit was developed .We used this established ELISA kit to detect the plasma concentration of OLC 1 protein in 281 NSCLC patients and 92 gender-and age-matched healthy controls .Area under the receiver operating characteristic curve (AUC) was used to evaluate the detection efficacy of OLC 1.Results We obtained 11 OLC1 monoclonal antibodies and successfully established the ELISA kit to detect the plasma concentration of OLC1 with a detection range from 1.95 ng/ml to 62.50 ng/ml.OLC1 concentration in the case group (124.69 ng/ml) was significantly higher than that in the control group (67.07 ng/ml, P<0.001).In the scenario of distinguishing NSCLC from control group , AUC result was 0.69.When the cut-off was set at 67.72 ng/ml, the sensitivity and specificity was 84.4%and 51.1%, respectively.In term of distinguishing early lung cancer (IA) from normal controls, the AUC, sensitivity and specificity were 0.68, 77.8% and 54.4%, respectively .Conclusion The plasma concentration of OLC 1 protein is significantly elevated in NSCLC patients.OLC1 may be as a potential cancer biomarker applied in clinical diagnosis .

Objective To explore the association between Toll-like receptor-8(TLR8) genetic polymorphisms and susceptibility to ischemic stroke (IS) in patients of different traditional Chinese medicine (TCM) pat-terns. Methods 311 IS patients of wind-phlegm stasis-obstruction pattern, 284 IS patients of qi-deficiency blood-stasis pattern and 605 controls were recruited. The real-time PCR technology was used to detect TLR8 gene expression levels. TLR8 rs3764880 single nucleotide polymorphism(SNPs) were genotyped using the Se-quenom Mass ARRAY platform. Levels of inflammatory cytokines were measured by using ELISA. Statistical analyses were carried out by using PLINK and SPSS 16. 0 software. Results TLR8 gene variant rs3764880 polymorphism was significantly associated with wind-phlegm stasis-obstruction pattern in IS males patients in the allele model (P<0. 05). In IS male patients of qi-deficiency blood-stasis pattern , level of IL-8 was high-er in the GG genotype carriers than the AA+AG genotype carriers (P<0. 05). The GG+AG genotype carri-ers had higher levels of TNF-α and IL-8 than the AA genotype carriers ( P <0. 05 ) . TLR8 gene variant rs3764880 polymorphism was significantly associated with the levels of TC (βb = -0. 03, Padj =0. 001) and LDL (βb= -0. 25, Padj =0. 018) in additive model in qi-deficiency blood-stasis pattern in Han Chinese IS male patients. Conclusion TLR8 gene rs3764880 polymorphism might be associated with the susceptibility to IS in patients of wind-phlegm stasis-obstruction and affect inflammatory reaction and lipid metabolism in Han Chinese males of qi-deficiency blood-stasis pattern.

Objective To investigate the relationship between tumor necrosis factor receptor-associated factor 6 ( TRAF6 ) gene polymorphisms and susceptibility to stroke with diverse TCM patterns and inflam-mation-related quantitative characters. Methods A case-control design was applied in this study. 311 stroke cases with the pattern of wind-phlegm stasis and 284 cases with qi-deficiency blood-stasis pattern were included in the case group, and 605 gender-matched, and age-matched cases were enrolled in the control group. The expression level of TRAF6 mRNA was determined by using real-time PCR ( RT-PCR), and two single nucleotide polymorphisms (SNP) of the TRAF6 gene, rs5030411 and rs5030416, were genotyped with Sequenom MassARRAY iPLEX platform. Enzyme-linked immuno sorbent assay ( ELISA) was applied to detect the serum levels of inflammatory cytokines. PLINK and SPSS software were used for statistical analysis. Results The TRAF6 mRNA expression level in patients with wind-phlegm stasis pattern or qi-deficiency blood-stasis pattern were significantly higher than those in the con-trol group ( P<0 . 050 ) . Rs5030416 polymorphism has a close association with susceptibility to stroke with wind-phlegm stasis pattern ( P =0 . 030 ) . No relationship was found between rs5030411 polymor-phism and stroke susceptibility ( P>0 . 050 ) . Patients carrying rs5030411-CT genotype in wind-phlegm stasis pattern group had remarkably higher interleukin-1b (IL-1b) and interleukin-6 (IL-6) levels than other genotype carriers. Moreover, compared with control group, the levels of IL-1b and IL-6 were signif-icantly higher in wind-phlegm stasis pattern group ( P<0 . 05 ) . Conclusion TRAF6 gene polymorphism rs5030416 may play a role in the occurrence of wind-phlegm stasis pattern, and rs5030411 polymorphism may engage in the inflammatory reaction in wind-phlegm stasis pattern.

The Gastroesophageal reflux disease(GERD)is related to the dynamic disorder of the digestive tract caused by the imbalance of the viscera and meridians.The spleen and stomach are the hub of the qi machinery,transforming the essence of water and valley into energy and regulating the Yin and Yang of all bodies.The sympathetic balance between Ren-Du and Qi is the motivity of the spleen and stomach.Interstitial cells of Cajal(ICC)are the hub of digestive motility,which can maintain mitochondrial energy metabolism through mitochondrial autophagy and improve the digestive motility.Therefore,in this paper,the molecular biological basis of GERD was discussed based on the"regulating pivot with pivot"theory that the pivots of viscera and meridians drive ICC mitochondrial energy balance.It also explains the feasibility of Qi-Shi-Sheng-Jiang-Gui-Yuan Decoction in treating GERD based on"regulating pivot with pivot",which is helpful to realize the microscopization and concretization of"regulating pivot with pivot"theory and realize the modernization of TCM theory.

Lung cancer is the malignant tumor with the highest mortality rate in the world. Heterogeneity of lung cancer, usually studied by sequencing technology, is considered to have important clinical significance in current studies. However, general sequencing technology can only explain the differences between samples integrally and its resolution is not enough to describe the differences between the individual cells. Therefore, people urgently hope to understand the cell type, state, subgroup distribution in the tumor microenvironment and the communication behavior between cells in the single cell level. Single-cell sequencing technology solves this problem. Using this technique will contribute to further understanding the mechanism of the occurrence and development of lung cancer, discovering new diagnostic markers and therapeutic targets, and providing theoretical references for the precise treatment of lung cancer patients in the future. This article reviews the progress of single-cell sequencing technology and focuses on its research on lung cancer tumor heterogeneity, tumor microenvironment, invasion and metastasis, treatment response, and drug resistance.
.

Ulcerative colitis（UC）is a disease characterized by chronic persistent inflammation of the colorectal mucosa. Its complex pathological mechanism is related to immune inflammation and enhanced apoptotic activity. The Janus kinase（JAK）/signal transducer and activator of transcription（STAT）is an important regulatory pathway in the body's physiological function, which can regulate the release of intestinal pro-inflammatory factors and induce apoptosis, resulting in colon tissue damage. In the condition of UC, the biological activities and expression levels of JAK and STAT increased, and the tissue inflammatory response and apoptosis rate increased, which led to the destruction of intestinal mucosal tissues. At present, in the treatment of UC, glucocorticoids and immunosuppressants are mainly employed to reduce intestinal inflammation. Although they can block the progress of UC to some extent, the adverse reactions are severe. A large number of studies have shown that traditional Chinese medicine（TCM） has significant advantages in the prevention and treatment of UC and can significantly reduce the recurrence rate of this disease. In recent years, plenty of studies have been carried out to explore the role of TCM in the treatment of UC by regulating the JAK/STAT pathway. The results have shown that the JAK/STAT pathway is the key target pathway of TCM in the treatment of UC. Based on the etiology and pathogenesis of deficiency and excess, TCM regulates the JAK/STAT pathway by clearing heat, drying dampness, cooling and activating blood, invigorating the spleen, warming the kidney, and performing both tonification and elimination to maintain the balance between pro-inflammatory factors and anti-inflammatory factors, weaken colonic inflammatory response, inhibit apoptosis, and play a role in the treatment of UC. The present study analyzed the mechanism and effect of TCM in intervening in UC by targeting the JAK/STAT signaling pathway and summarized the molecular mechanisms of different cytokines such as interleukin-6（IL-6）, IL-10, IL-23, microRNA（miRNA）-146a, and suppressors of cytokine signaling 2/3（SOCS2/3） on many family subtypes of the JAK/STAT signaling pathway to facilitate the comprehensive understanding of researchers on the mechanism of TCM on the JAK/STAT pathway in UC, which is expected to provide a theoretical basis for the treatment of UC and further drug development.

BACKGROUND: Lung cancer is the most common malignancy world-wide. There are a variety of immune infiltrating cells in tumor microenvironment, which is an important component of tumor immunity and has clinical significance for the prognosis of patients. CD45RO is a surface marker of memory T cells. The expression of CD45RO⁺ tumor infiltrating lymphocytes (TILs) is associated with the prognosis of many tumors. The purpose of this study was to evaluate the relationship between the density of CD45RO⁺ TILs in tumor and stromal area and the clinical characteristics of patients with non-small cell lung cancer (NSCLC) and its impact on the prognosis of patients. We aimed to explore the clinical value of CD45RO⁺ TILs and programmed cell death ligand 1 (PD-L1) as prognostic markers. METHODS: Multiple fluorescent immunohistochemical staining was used to stain the tissue microarray chips of 167 patients with NSCLC, marking CD45RO, cytokeratin (CK) and PD-L1. Using artificial intelligence image recognition technology and tumor cell-specific CK staining, divide the tumor and stromal area in the tissue, evaluate the density of CD45RO⁺ TILs in the tumor and stromal area, and the expression level of PD-L1 in tumor cells. The non-parametric test was used to analyze the relationship between CD45RO⁺ TILs and the clinical characteristics of patients, and the Kaplan-Meier method and Cox risk ratio model were used to analyze the relationship between CD45RO⁺ TILs independently or in combination with PD-L1 and tumor prognosis. RESULTS: The density of CD45RO⁺ TILs was significantly associated with patient age, smoking, tumor stage, and pathological type. Single-factor survival analysis showed that NSCLC (P=0.007) stromal region and lung adenocarcinoma (LUAD) (P<0.001) with CD45RO⁺ TILs high density had better OS. Multivariate survival analysis showed that the high density of CD45RO⁺ TILs in the stromal region of NSCLC (HR=0.559, 95%CI: 0.377-0.829, P=0.004) and lung adenocarcinoma (HR=0.352, 95%CI: 0.193-0.641, P=0.001) were independent prognostic factors for overall survival time (OS). Combined with PD-L1 score of tumor cells in tumor tissues and infiltration score of CD45RO⁺ TILs in all tumor tissues, the patients were divided into 4 groups: patients with PD-L1⁺/CD45RO⁺ had the longest disease-free survival (DFS) time, and patients with PD-L1⁺/CD45RO- had the shortest DFS time. Multivariate Cox regression analysis showed that PD-L1⁺/CD45RO- was an independent prognostic factor for DFS and had a higher risk of poor prognosis compared to the other three groups (HR=2.221, 95%CI: 1.258-3.919, P=0.006). CONCLUSIONS In tumor tissues, the density of CD45RO⁺ TILs, as well as the combination of CD45RO⁺ TILs and PD-L1 in tumor areas, significantly correlated with clinicopathological features and prognosis of NSCLC, which can be used as a new prognosis marker.