Objective:To explore effects of T helper type 1 cells (Th1) to T helper type 2 cells (Th2) ratio and the related cytokines on the prognosis of patients with colorectal neoplasms.Methods:A total of 98 colorectal neoplasms patients undergoing the surgery admitted in Suqian Hospital Affiliated to Xuzhou Medical University from December 2015 to December 2017 were enrolled, and all patients were selected as the colorectal cancer group. According to Dukes staging criteria, patients were divided into stage A (25 cases), stage B (30 cases) and stage C (43 cases). In addition, 72 healthy subjects who underwent physical examination in Suqian Hospital Affiliated to Xuzhou Medical University during the same period were selected as the healthy control group. Preoperative venous blood on an empty stomach was extracted from the healthy control group and the colorectal cancer group. Flow cytometry was used to analyze the Th1/Th2 ratio in peripheral blood. The levels of cytokines interferon (IFN)-γ, interleukin (IL)-2, IL-4 and IL-10 in serum samples were detected by using enzyme-linked immunosorbent assay (ELISA). After operation, patients were followed up by telephone or outpatient service. The Th1/Th2 ratio and levels of IFN-γ, IL-2, IL-4 and IL-10 of both groups at different stages of both groups were compared. The correlation between Th1/Th2 ratio and the clinicopathological characteristics of colorectal cancer patients was analyzed. Kaplan-Meier method was used to make survival analysis and Cox regression model was used to analyze influencing factors for overall survival (OS).Results:The Thl/Th2 ratio in colorectal cancer patients was lower than that in the healthy control group (5.13±2.04 vs. 11.82±2.76, t = 18.177, P < 0.01). The lymphovascular invasion and Dukes stage C ratio in patients with decreased Th1/Th2 ratio were higher than those in patients with increased Th1/Th2 ratio ( χ2 values were 16.403, 16.248, both P < 0.01). The levels of IFN-γ and IL-2 in serums of colorectal patients were (95±15) ng/L and (78±10) ng/L, respectively, which were lower than those in the healthy control group [(157±17) ng/L and (123±12) ng/L, t values were 25.160, 26.622, all P < 0.01]. The levels of IL-4 and IL-10 in the colorectal cancer group were (87±16) ng/L and (178±18) ng/L, respectively, which were higher than those in the healthy control group [(46±9) ng/L and (124±12) ng/L] ( t values were 19.577, 22.095, all P < 0.01). The follow-up time ranged from 31.0 to 55.0 months, and the median follow-up time was 37.2 months and the median OS time was 21.0 months. Survival analysis showed that the OS of patients with increased Th1/Th2 ratio was better than that of patients with reduced Th1/Th2 ratio ( χ2 = 7.287, P = 0.007). Multivariate Cox regression analysis showed that lymph node metastasis, tumor stage, and Th1/Th2 ratio were independent influencing factors for OS in colorectal cancer patients ( OR values were 8.541, 3.442, 1.275, all P < 0.05). Conclusion:The imbalance of related cytokines secreted by Th1 and Th2 cells and the decrease in the ratio of Th1/Th2 are related to the progression and the poor prognosis of colorectal cancer.

Objective:To analyze the risk factors of first-episode hematogenous metastasis in patients with thoracic esophageal squamous cell carcinoma (ESCC) who received non-surgical treatment after radiotherapy and chemotherapy, and its impact on survival and prognosis.Methods:The clinical data of 230 ESCC patients who met the inclusion criteria and received radical radiotherapy in Tengzhou Central People′s Hospital and Affiliated Hospital of Xuzhou Medical University from January 2011 to October 2018 were retrospectively analyzed.Logistic regression analysis and survival were used to analyze the risk factors and prognosis of blood group metastasis after treatment.Results:In 230 patients with thoracic esophageal cancer, 70 cases (30.4%) developed hematogenous metastasis for the first time.Compared with patients without hematogenous metastasis, the median overall survival was 15 months and 20 months (χ 2=7.249, P=0.007), and the median progression free survival was 9 months and 13 months (95% CI was 7.2-10.8 months and 10.8-15.2 months, respectively χ 2=21.664, P<0.001). Logistic multivariate analysis showed that there was significant difference in the occurrence of hematogenous metastasis among different N stages (χ 2=30.764, P<0.001). N stage was an independent factor for judging hematogenous metastasis, and the increased N stage increased the risk of hematogenous metastasis (OR value were 6.000, 12.629 and 48.167, respectively; 95% CI were 1.712-21.025, 3.546-44.976 and 10.848-213.858, respectively; all P<0.05). The overall survival time of patients with concurrent chemoradiotherapy before hematogenous metastasis was longer than that of patients with sequential chemoradiotherapy and radiotherapy alone (χ 2=10.002, P=0.007). Stratified analysis showed that adjuvant chemotherapy after concurrent chemoradiotherapy could prolong the overall survival of patients with N2 and N3 (χ 2=11.025, P=0.001). Conclusion:N staging is an independent factor to judge the hematogenous metastasis.ESCC patients with hematogenous metastasis after chemoradiotherapy have poor prognosis.N2, N3 patients with concurrent chemoradiotherapy after adjuvant chemotherapy have clinical benefits.

Objective To study the clinical efficiency and adverse reactions of simultaneous modulated accelerated radiotherapy (SMART)and intensity-modulated radiation therapy (IMRT)in advanced cervical cancer. Methods Sixty patients with advanced cervical cancer were collected from April 2011 to April 2017 in our hospital. The 60 patients were randomly divided into experimental group (30 cases)and control group (30 cases)by using stratified randomization method. The two groups were given intracavitary irradiation and concur-rent chemotherapy. The patients in experimental group were treated with SMART and the patients in control group were treated with IMRT. 95％ planned target volume was 50. 4 Gy/ 28 F in the two groups and the dose for IMRT with simultaneous integrated boost was 64. 4 Gy/ 28 F to the planning target volume. Disease progres-sion,survival time and adverse reactions of the two groups were compared. Results At the end of radiothe-rapy,the experimental group had 23 patients with complete response (CR),4 patients with partial response (PR),2 patients with unaltered stable disease (SD),1 patient with progressive disease (PD),and the control group had 22 patients with CR,3 patients with PR,3 patients with SD,2 patients with PD. The overall effi-ciency of the experimental group was slightly higher than that of the control group (90. 0％ vs. 83. 3％),but the difference was not statistically significant (χ2 = 0. 144,P = 0. 704). After 3 months of radiotherapy,the experimental group had 28 patients with CR,1 patient with PR,1 patient with PD,and the control group had 22 patients with CR,2 patients with PR,3 patients with SD,3 patients with PD. The overall efficiency of the experimental group (96. 7％)was higher than that of the control group (96. 7％ vs. 80. 0％),but the diffe-rence was not statistically significant (χ2 = 2. 588,P = 0. 108). The median overall survival time of the experi-mental group and control group were 43 months and 38 months,and the difference was statistically significant (χ2 = 7. 087,P = 0. 008). The 1-year survival rates of the two groups were 96. 6％ and 85. 7％,and the 3-year survival rates were 86. 2％ and 60. 7％,respectively. There were no significant differences in the inci-dences of gastrointestinal reaction (66. 7％ vs. 63. 3％,χ2 = 0. 073,P = 0. 787),urinary system reaction (33. 3％ vs. 30. 0％,χ2 = 0. 077,P = 0. 781)and bone marrow suppression (83. 3％ vs. 86. 7％,χ2 =0. 000,P = 1. 000)between the two groups. Conclusion The efficiency of advanced cervical cancer patient treated with SMART is better than IMRT,and the adverse reactions are tolerable,which is worthy of clinical promotion.

Super hydrophobic interface modified with silver nanoparticles was fabricated for the detection of pesticide residues.By using a chemical reduction method,silver nanoparticles were deposited on the substrate surfaces with different microscopic pore structures.Two kinds of composite substrates,including regular stainless steel mesh and cellulose polyester film,were used.The pre-treatment of the substrate with fluoridated reagents was used to form a super hydrophobic interface,which made the target molecules on the surface concentrate effectively.The surface with the cellulose polyester substrate was used to detect Rhodamine 6G (R 6G) effectively with surface enhanced Raman scattering (SERS) technique.The results showed that the detection hmit was 10-16 mol/L.In addition,the surfaces based on the stainless steel mesh and cellulose polyester substrate were used to detect trichlorfon pesticide with detection limits of 1 × 10-15 mol/L and 1 × 10-16 mol/L,respectively.

HCV-related decompensated cirrhos,hypersplenism,thrombocytopenia,which not only affect the standard antiviral therapy,fail to achieve the sustained virological response(SVR),but also increase the risk of infection and bleeding.The only successful option is liver transplantation (LT),but the recurrence of HCV after LT remains to be resolved.The patients of HCV genotype 2 are suitable for splenectomy and antiviral therapy following splenectomy,which can achieved a higher SVR and reversed cirrhosis.As an effective alternative to splenectomy,the partial splenic embolization (PSE) can improve the changes of portal hemodynamics and reduce the sequelae of portal hypertension.The appearance of direct antiviral drugs (DAAs)has bring hope for those with decompensated cirrhos and whom IFN is contraindicated or tolerated poorly,those who are waiting for LT or with recurrence of hepatitis C after LT.The treatment of patients with decompensated cirrhos is as follows.

Background and purpose:The expression ofRab11 gene was increased incervical cancer cell and may be involved in the cellular malignant transformation. This study used the sequence-speciifc siRNA knocking down the expression of Rab11 gene and aimed to investigate its effect on invasion and migration of cervical cancer cell lines HeLa/SiHa and its mechanism.Methods:HeLa/SiHa cells were divided into 2 groups: non-speciifc siRNA group transfected with unrelated siRNA (Rab11-NC) and Rab11 siRNA group transfected with Rab11 siRNA (Rab11siRNA). Western blot was used to examine the Rab11 protein expression. Cell migration and invasion were detected by cell scratch and Transwell invasion assay. Western blot was used to further investigate the expression of Rac1, matrix metal-loproteinase 2 (MMP2) and MMP9 which were critical for regulating cell invasion. Moreover, immunolfuorescence was used to identify intracellular location of Rac1 in HeLa/SiHa cells.Results:The Rab11 siRNA inhibited expression of Rab11 gene (P<0.01). The invasion and migration capacities of HeLa/SiHa cells were markedly inhibited in Rab11siR-NA group (P<0.05). The expression of Rac1 signiifcantly decreased (P<0.01). The expression of MMP2 and MMP9 de-creased (P<0.05) as well. The recruitment of Rac1 to protruding edge signiifcantly decreased following down-regulation of Rab11.Conclusion:Down-regulatedRab11 expression could inhibit the expression of Rac1, MMP2 and MMP9, and alter the location of Rac1, leading to suppression of HeLa/SiHa cells migration and invasion.

Objective To explore the expression of Ras-related protein 11(Rab11)in hypoxia, the effect of Rab11 on the invasion and migration of cervical cancer cell line SiHa and its possible mechanism. Methods SiHa cells were divided into 4 groups, the normoxic blank group (normal culture in normoxia), the hypoxic blank group (normal culture in hypoxia), the negative control group [transfection of negative control small interfering RNA(siRNA)in hypoxia], the Rab11-siRNA group (transfection of Rab11 siRNA in hypoxia). Western blot was used to examine the expression of Rab11, integrin α5, integrin β3, phosphorylated focal adhesion kinase(p-FAK), phosphorylated phosphatidylinositol 3 kinase(p-PI3K) protein, together with the expression of Ras correlative C3 creotoxin substrate 1(Rac1), which was critical in regulating cell invasion. The mRNA expression of Rab11 in the 4 groups was detected by realtime-qPCR. The cell invasion was detected by matrigel assay, while the cell migration was detected by transwell assay. Immunofluorescence was used to identify intracellular location of Rac1 in SiHa cell. Results (1) The expression of Rab11, intergrin α5, intergrin β3, p-FAK, p-PI3K and Rac1 in the normoxic blank group were 0.56±0.04, 0.33±0.03, 0.32±0.03, 0.36±0.03, 0.35±0.03 and 0.47±0.03, respectively. In the hypoxic blank group, they were 0.73±0.03, 0.74±0.03, 0.61±0.03, 0.62±0.03, 0.60±0.03 and 0.73±0.03, respectively. In the negative control group, their expressions were 0.72±0.03, 0.73±0.03, 0.59±0.03, 0.61±0.03, 0.59±0.03 and 0.72±0.03, respectively. While in the Rab11-siRNA group, they were 0.44±0.03, 0.30±0.03, 0.29±0.03, 0.30±0.03, 0.30±0.03 and 0.34±0.04, respectively. The expressions of Rab11, α5, β3, p-FAK, p-PI3K and Rac1 were significantly higher in the hypoxic blank group than in the normoxic blank group(P<0.05), and were significantly lower in the Rab11-siRNA group than in the hypoxic blank group and the negative control group(P<0.05). (2) The expressions of Rab11-mRNA were 1.000±0.000, 1.454±0.114, 1.442±0.101, 0.570± 0.046 in the normoxic blank group, the hypoxic blank group, the negative control group and the Rab11- siRNA group, respectively. It was significantly higher in the hypoxic blank group than in the normoxic blank group(P<0.05), and was significantly lower in the Rab11-siRNA group than in the hypoxic blank group and the negative control group(P<0.05). (3) By Matrigel, the invasion cell number in the normoxic blank group, the hypoxic blank group,the negative control group and the Rab11-siRNA group were 65±12, 106±16, 104± 17 and 50±11, respectively. The invasion capacity was significantly higher in the hypoxic blank group than in the normoxic blank group(P<0.05), and was significantly lower in the Rab11- siRNA group than in the hypoxic blank group and the negative control group(P<0.05). (4) By transwell assay, the migration cells in the normoxic blank group, the hypoxic blank group, the negative control group and the Rab11-siRNA group were 127±12, 169±15, 161±13 and 77±13, respectively. The capacity of invasion was significantly higher in the hypoxic blank group than in the normoxic blank group(P<0.05), and was significantly lower in the Rab11- siRNA group than in the hypoxic blank group and the negative control group(P<0.05). (5) The immunofluorescence showed that the red fluorescence intensity around nucleus was significantly increased in the normoxic blank group, the hypoxic blank group and the negative control group than in the Rab11- siRNA group. Conclusions Hypoxia could promote the invasion and migration of SiHa cells. In hypoxia, the down regulation of Rab11 expression could inhibit the invasion and migration of SiHa cells. This might be due to the decreased expression of the intergrin α5, intergrin β3, p-FAK, p-PI3K and Rac1 protein.

Objective To explore the effects of Rab11 on biological functions of human cervical cancer cell line HeLa through regulating the expression levels of Rab11. Methods The Rab11 siRNA was transfected into HeLa cells and the expression of Rab11 was detected by Western blot. CCK8 assay, colony formation experiments, EdU assay and Transwell assay were adopted to observe the effect of Rab11 on HeLa cells proliferation and invasion. Results The expression of Rab11 was decreased significantly in HeLa cells transfected with Rab11 siRNAs than that in control siRNA (1.096 ±0.091 vs 1.735 ±0.084, P< 0.01). The proliferation was markedly inhibited in Rab11 siRNA group compared with that in control siRNA group (48 h:0.721±0.092 vs 1.090±0.099; 72 h: 0.956±0.105 vs 1.482±0.096; 96 h: 1.231±0.099 vs 1.720±0.174, P< 0.01), the number of colonies was lower than that in control siRNA group (36±1 vs 75±8, P< 0.01) and so was proliferation rate [(33.880±1.902) % vs (45.570±2.025) %, P< 0.05]. The cell invasion rate of Rab11 siRNA group was lower than that of control siRNA group [(38.6 ±0.8) % vs (100.0 ±0.2) %, P< 0.01]. Conclusion Down-regulation of Rab11 expression can inhibit the growth of HeLa cells.

Objective To compare the results of anti-cardiolipin (ACA)measuring by two commercial available kits.Methods ACA in total of 66 serum samples were both determined by kits from Euroimmun and YHLO simultaneously,then the re-sults were analyzed comparatively and correlatively.The Euroimmun kit was applied to determination the level of ACA-IgA/G/M,and the YHLO kit determined ACA-IgG/M and anti-β2-glycoprotein I antibody (β2GPI IgG).Results The positive rate by Euroimmun kit was 37.88% (25/66),while 31.82% (21/66)was positive (one positive among ACA-IgG,IgM andβ2GPI IgG)when determined by YHLO kit,and there was no significant difference between the two kits.The accordance rate of the two kits was 87.88% (58/66).The ACA-IgA/G/M value by Euroimmun kit and the summation of ACA-IgG, IgM andβ2GPI IgG by YHLO kit showed well linear correlation (r 2 =0.892,P <0.01).Conclusion Results from the two kits were consistent and correlated well,and they are suitable for the clinical application;these two kits have their own char-acteristics,which could be used by individual or combination accordingly.

Objective To establish radiation?resistant lung carcinoma cell lines, and to investigate the changes in morphology, apoptosis, invasive migration, and epithelial?mesenchymal transition ( EMT) in cells. Methods The radiation?resistant lung carcinoma cell lines were obtained by exposure of lung carcinoma cell lines, A549 and H1299, to radiation with a low dose in fractions, a sublethal dose, or a gradually increasing dose. The morphological changes in cells, radiosensitivity, survival rates after exposure, apoptosis rates, changes in invasive migration, and expression of EMT marker proteins were evaluated using microscopy, colony formation assay, CCK?8 assay, flow cytometry, transwell migration assay, and Western blot, respectively. Results Radiation with a gradually increasing dose successfully induced the radiation?resistant cell lines, A549R and H1299R. The morphological study showed that the morphology of radiation?resistant cells was converted to the morphology of mesenchymal cells. Compared with A549 and H1299 cells, the values of D0 , Dq , and SF2 were significantly increased in A549R ( P=0.017,P=0.001,P=0.000) and H1299R (P=0.033,P=0.000,P=0.008) cells, respectively;the values of α and α/β were significantly reduced in A549R (P=0.018;P=0.007) and H1299R (P=0.001;P=0.009) cells, respectively. The survival rates in A549R and H1299R cells after exposure to radiation with various doses were significantly higher than those in the control groups (all P<0.05). After exposure, the apoptosis rates were significantly reduced in A549R and H1299R cells ( P=0.02,P=0.01);the invasion and migration rates were significantly increased in A549R (P=0.000;P=0.001) and H1299R (P=0.001,P=0.002) cells;the expression of E?cadherin was significantly down?regulated in A549R and H1299R cells (P=0.00,P=0.01), while the expression of vimentin was significantly elevated in A549R and H1299R cells ( P= 0. 02, P= 0. 01 ) . Conclusions The radiation?resistant lung carcinoma cell lines are successfully established. Both cell lines show enhanced invasion and migration, which may be associated with EMT.