Objective To study tumor immune escape in a gastric carcinoma cell line expressing human indoleamine 2,3-dioxygenase(IDO).Methods Human IDO gene was cloned by RT-PCR and the vector for pIRES_2-EGFP-IDO was constructed.BGC-823 cells were transfected with the plasmid using eleetroporation.The integrated INDO genes were detected by RT-PCR and Western blot.The enzyme activity of IDO were measured.T cells from gastric cancer patients were cecuhured with BGC-823 transfected with IDO or added with 1-MT circumstance,T cell-mediated cytotoxicity and proliferation were detected.Results Higher level expression of IDO mRNA and IDO protein Was detected in tumor cells transfected with IDO gene.The level of kynurenic acid was higher in transfected cells compared with no-transfeeted group (4.84±0.11)mg/L vs.(1.83±0.10)mg/L,P=0.000.The cytotoxicity ratio of the IDO transfected group and transfected group with 1-MT circumstance (1-MT group) was lower than control group (P<0.05).The inhibition rate of transfected group with 1-MT group Was higher than control group(P<0.05).Conclusion Gastric cancer cell lines encoded with IDO inhibits T cell-mediated cytotoxicity and proliferation.

Objective To investigate the effects and mechanisms of Omega-3 polyunsaturated fatty acid (ω-3 PUFA) supplementation on spatial learning and memory capacity,neuronal apoptosis,microglia activation,neuroinflammatory response and nuclear factor-κB (NF-κB) pathway in cognitive impairment model rats.Methods Cognitive impairment model rats were induced by intraperitoneal injection of D-galactose.Thirty-six Wistar rats were randomly divided into three groups:control group (Con),Cognitive impairment group (AD) and ω-3 PUFA supplementation group (AD+ω-3).The spatial learning and memory capacity of experimental rats were examined by the Morris water maze (MWM).Apoptotic neurons were determined by Nissl staining.The microglia activation relatived protein (Iba-1) expressions was determined by immunohistochemistry staining and western blot,respectively.While,the serum levels of tumor necrosis factor-α (TNF-α),interleukin (IL)-1 and IL-6 were tested by enzyme linked immunosorbent assay (ELISA).In addition,the protein expression of NF-κB related indexes including NF-κB p65,p-IκB and TLR4 were tested using western blot.Results Compared with the Con group,the escape latency increased and the distance percentage in target quadrant decreased;neurons apoptosis,microglia activation and neuroinflammatory factors significantly increased in the other two groups (P=0.00).Compared with the AD group,the escape latency was remarkably shorter (2 d:41.35±2.34vs.58.07±3.27,P=0.03;3 d:35.07±2.45 vs.44.39±3.21,P=0.02;4d:28.12±2.43vs.35.63±2.20,P=0.01;5 d:23.74±1.06 vs.29.76±1.15,P=0.03),and the distance percentage in target quadrant increased significantly [(48.26±4.02)％ vs.(34.14±3.49)％,P =0.01] after ω-3 PUFA supplementation.The incidence of neurons apoptosis significantly decreased in AD + ω-3 group [(29.93 ±3.05) ％ vs.(47.58±4.14)％,P=0.01].ω-3 PUFA supplementation inhibited the protein expression of microglia,and meanwhile inhibed the expression of microglia-induced neuroinflammatory factors [TNF-α:(85.27±9.77vs.156.13±14.53) pg/ml,P=0.00;IL-1:(41.23±5.38vs.75.04±9.27) pg/ml,P=0.01;IL-6:(47.58±4.23 vs.97.47±9.09) pg/ml,P=0.00].Compared with AD group,the protein expression of NF-κB pathway significantly decreased in AD+ω-3 group.Conclusion ω-3 PUFA supplementation can inhibited microglia activation,decrease microglia-mediated neuroinflammatory response,reduce neuron apoptosis and markedly improve spatial learning and memory capacity in cognitive impairment rats,possibly mediated by inhibiting NF-κB pathway.

OBJECTIVETo investigate the best diagnostic and therapeutic method for primary lymphoma of the spleen.

METHODSClinicopathologic features of 23 patients treated from January 1956 to August 1999 were analyzed retrospectively.

RESULTSAll patients but one for exploration only (96%) underwent resection of the tumor. They accepted chemotherapy after operation. 23 patients were confirmed pathologically. B-cell type non-Hodgkin's lymphoma was noted in 21 patients and T-cell letion in 2. According to Ahman's staging, 9 patients belonged to stage I, 8 stage II, and 6 stage III. The 5-year survival rates were 50%, 40% and 16% respectively.

CONCLUSIONSThe diagnosis of splenic lymphoma is dependent mainly on B-ultrasound examination and CT scanning. Splenectomy combined with chemotherapy may provide optimum therapy for patients with splenic lymphoma.

Adult ; Aged ; Combined Modality Therapy ; Drug Therapy ; Female ; Humans ; Lymphoma ; diagnosis ; drug therapy ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Splenectomy ; Splenic Neoplasms ; diagnosis ; drug therapy ; surgery

Objective:To investigate the role and the mechanism of Ras-associated binding protein23 (RAB23) in the migration and invasion of esophageal squamous cell carcinoma (ESCC) cells.Methods:RAB23 mRNA levels were measured in 16 pairs of ESCC and adjacent normal tissues via real-time polymerase chain reactions. RAB23 mRNA levels in the ESCC and adjacent normal tissues of dataset GSE20347 deposited in the Gene Expression Omnibus (GEO) database were also analyzed. Immunohistochemistry (IHC) was used to detect the RAB23 protein expressions in 106 pairs of ESCC and adjacent normal tissues, as well as in the lymph glands and primary tumor tissues of 33 patients with positive lymph nodes and 10 patients with negative lymph nodes. Endogenous RAB23 expression was transiently depleted using siRNAs (si-NC, si-RAB23-1, and si-RAB23-9) or stably reduced using shRNAs (sh-NC and sh-RAB23) in ESCC KYSE30 and KYSE150 cells, and the knockdown efficiency was tested using Western blot assays. Cell counting kit-8 assays and mouse xenograft models were used to test the proliferation of ESCC cells . Transwell assays and tail vein-pulmonary metastasis models in immunocompromised mice were used to examine the migration and invasion of ESCC cells. Cell adhesion assays were used to test the adhesion of ESCC cells. RNA-seq assays were used to analyze how RAB23 knockdown influenced the expression profile of ESCC cells and the implicated signal pathways were confirmed using Western blot assays. Results:The RAB23 mRNA expression in 16 cases of ESCC tissues was 0.009 7±0.008 9, which was markedly higher than that in adjacent normal tissues (0.003 2±0.003 7, P=0.006). GEO analysis on RAB23 expressions in ESCC and adjacent normal tissues showed that the RAB23 mRNA level in ESCC tissues (4.30±0.25) was remarkably increased compared with their normal counterparts (4.10±0.17, P=0.037). Among the 106 pairs of ESCC and tumor-adjacent normal tissues, 51 cases exhibited low expression of RAB23 and 55 cases showed high expression of RAB23, whereas in the paired tumor-adjacent normal tissues 82 cases were stained weakly and 24 strongly for RAB23 protein. These results indicated that RAB23 expression was markedly increased in ESCC tissues ( P<0.001). Additionally, only 1 out of 33 primary ESCC tissues with positive lymph nodes showed low RAB23 protein expression. On the other hand, 7 samples of primary ESCC tissues with negative lymph nodes were stained strongly for RAB23 while its level in the other 3 samples was weak. These results showed that RAB23 expression was remarkably increased in primary ESCC tissues with positive lymph nodes compared with those with negative lymph nodes ( P=0.024). Further tests showed that 32 out of 33 positive lymph nodes were stained strongly for RAB23, whereas no negative lymph nodes ( n=10) exhibited high expression of RAB23 ( P<0.001). Both transient and stable knockdown of endogenous RAB23 expression failed to cause detectable changes in the proliferation of KYSE30 cells in vitro and in vivo, but attenuated the migration and invasion of KYSE30 cells as well as the invasion of KYSE150 cells. RAB23 knockdown was found to significantly decrease the number of adhesive KYSE30 cells in the sh-RAB23 group (313.75±89.34) compared with control cells in the sh-NC group (1 030.75±134.29, P<0.001). RAB23 knockdown was also found to significantly decrease the number of adhesive KYSE150 cells in the sh-RAB23 group (710.5±31.74) compared with the number of control cells in the sh-NC group (1 005.75±61.09, P<0.001). RNA-seq assays demonstrated that RAB23 knockdown using two siRNAs targeting RAB23 mRNA markedly impaired focal adhesion-related signal pathways, and decreased the levels of phosphorylated FAK (p-FAK) and phosphorylated paxillin (p-paxillin) in KYSE30 and KYSE150 cells. Conclusions:Significantly increased RAB23 in ESCC tissues positively correlates with lymph node metastasis. Depleted RAB23 expression attenuates focal adhesion-related signal pathways, thus impairing the invasion, metastasis, and adhesion of ESCC cells.

Objective:To investigate the role and the mechanism of Ras-associated binding protein23 (RAB23) in the migration and invasion of esophageal squamous cell carcinoma (ESCC) cells.Methods:RAB23 mRNA levels were measured in 16 pairs of ESCC and adjacent normal tissues via real-time polymerase chain reactions. RAB23 mRNA levels in the ESCC and adjacent normal tissues of dataset GSE20347 deposited in the Gene Expression Omnibus (GEO) database were also analyzed. Immunohistochemistry (IHC) was used to detect the RAB23 protein expressions in 106 pairs of ESCC and adjacent normal tissues, as well as in the lymph glands and primary tumor tissues of 33 patients with positive lymph nodes and 10 patients with negative lymph nodes. Endogenous RAB23 expression was transiently depleted using siRNAs (si-NC, si-RAB23-1, and si-RAB23-9) or stably reduced using shRNAs (sh-NC and sh-RAB23) in ESCC KYSE30 and KYSE150 cells, and the knockdown efficiency was tested using Western blot assays. Cell counting kit-8 assays and mouse xenograft models were used to test the proliferation of ESCC cells . Transwell assays and tail vein-pulmonary metastasis models in immunocompromised mice were used to examine the migration and invasion of ESCC cells. Cell adhesion assays were used to test the adhesion of ESCC cells. RNA-seq assays were used to analyze how RAB23 knockdown influenced the expression profile of ESCC cells and the implicated signal pathways were confirmed using Western blot assays. Results:The RAB23 mRNA expression in 16 cases of ESCC tissues was 0.009 7±0.008 9, which was markedly higher than that in adjacent normal tissues (0.003 2±0.003 7, P=0.006). GEO analysis on RAB23 expressions in ESCC and adjacent normal tissues showed that the RAB23 mRNA level in ESCC tissues (4.30±0.25) was remarkably increased compared with their normal counterparts (4.10±0.17, P=0.037). Among the 106 pairs of ESCC and tumor-adjacent normal tissues, 51 cases exhibited low expression of RAB23 and 55 cases showed high expression of RAB23, whereas in the paired tumor-adjacent normal tissues 82 cases were stained weakly and 24 strongly for RAB23 protein. These results indicated that RAB23 expression was markedly increased in ESCC tissues ( P<0.001). Additionally, only 1 out of 33 primary ESCC tissues with positive lymph nodes showed low RAB23 protein expression. On the other hand, 7 samples of primary ESCC tissues with negative lymph nodes were stained strongly for RAB23 while its level in the other 3 samples was weak. These results showed that RAB23 expression was remarkably increased in primary ESCC tissues with positive lymph nodes compared with those with negative lymph nodes ( P=0.024). Further tests showed that 32 out of 33 positive lymph nodes were stained strongly for RAB23, whereas no negative lymph nodes ( n=10) exhibited high expression of RAB23 ( P<0.001). Both transient and stable knockdown of endogenous RAB23 expression failed to cause detectable changes in the proliferation of KYSE30 cells in vitro and in vivo, but attenuated the migration and invasion of KYSE30 cells as well as the invasion of KYSE150 cells. RAB23 knockdown was found to significantly decrease the number of adhesive KYSE30 cells in the sh-RAB23 group (313.75±89.34) compared with control cells in the sh-NC group (1 030.75±134.29, P<0.001). RAB23 knockdown was also found to significantly decrease the number of adhesive KYSE150 cells in the sh-RAB23 group (710.5±31.74) compared with the number of control cells in the sh-NC group (1 005.75±61.09, P<0.001). RNA-seq assays demonstrated that RAB23 knockdown using two siRNAs targeting RAB23 mRNA markedly impaired focal adhesion-related signal pathways, and decreased the levels of phosphorylated FAK (p-FAK) and phosphorylated paxillin (p-paxillin) in KYSE30 and KYSE150 cells. Conclusions:Significantly increased RAB23 in ESCC tissues positively correlates with lymph node metastasis. Depleted RAB23 expression attenuates focal adhesion-related signal pathways, thus impairing the invasion, metastasis, and adhesion of ESCC cells.

OBJECTIVETo investigate the clinicopathological characteristics, diagnosis, treatment and prognosis of patients with primary gastric adenosquamous cell carcinoma.

METHODSA total of 5 562 patients with gastric neoplasm were admitted in Tianjin Medical University Cancer Institute and Hospital from January 2001 to January 2011. Among them 42 patients were diagnosed as primary gastric adenosquamous cell carcinoma, accounting for 0.76% of all the patients. The clinicopathological and follow-up data of these 42 patients with primary gastric adenosquamous cell carcinoma were retrospectively analyzed, and Cox proportional hazard model was used to analyze the prognostic factors of gastric adenocarcinoma squamous cell carcinoma.

RESULTSAmong above 42 patients, 32 were male and 10 were female, with a male-to-female ratio of 3.2/1.0 and the average age was 63 years (range: 46 to 77 years). Five patients (11.9%) were confirmed as adenosquamous cell carcinoma by preoperative pathological examination, while other 37 patients were diagnosed as adenocarcinoma preoperatively. According to the 7th edition AJCC TNM classification system for gastric adenocarcinoma, 5 patients (11.9%) were in stage II(, 30 patients (71.4%) in stage III( and 7 patients (16.7%) in stage IIII(. The maximum tumor diameter was > 5 cm in 18 patients (42.9%). Borrmann type III(-IIII( was found in 29 patients (69.0%), and poorly differentiated (or undifferentiated) tumor was found in 32 patients (76.2%). Radical operations were performed in 31 patients (73.8%), the reasons of non radical operations included infiltration of pancreas in 3 patients, infiltration of radices mesocili transvers in 1 patient and classification of stage IIII( in 7 patients. Lymph node dissection was performed in 37 patients, 83.8% of them (31/37) was found with lymphatic metastases. Twenty-five patients received adjuvant chemotherapy except for 7 patients in stage IIII( and 10 patients who refused adjuvant chemotherapy. All the patients had an average survival time of 36.4 months and median survival time of 28.0 months, and the overall 1-, 3- and 5-year survival rates were 82.2%, 42.3% and 18.2% respectively. Univariate analysis revealed that tumor size (χ=4.039, P=0.044), Borrmann type (χ=18.728, P=0.000), tumor differentiation (χ=19.612, P=0.000), radical gastectomy (χ=41.452, P=0.000), lymph node metastasis (χ=9.689, P=0.002) and clinical stage (χ=26.277, P=0.000) were associated with postoperative survival. Multivariate analysis revealed that tumor differentiation (HR=10.560, 95%CI:2.263-49.281, P=0.003), radical gastrectomy (HR=4.309, 95%CI:1.311-14.168, P=0.016) and clinical stage (HR=2.392, 95%CI:1.022-5.600, P=0.044) were independent prognosis factors.

CONCLUSIONSPrimary gastric adenosquamous cell carcinoma is rare with poor prognosis. Radical gastrectomy is recommended. Tumor differentiation, radical gastrectomy and clinical stage are important indicators to evaluate prognosis of primary gastric adenosquamous cell carcinoma.

Adenocarcinoma ; diagnosis ; mortality ; pathology ; therapy ; Aged ; Carcinoma, Squamous Cell ; diagnosis ; mortality ; pathology ; therapy ; Chemotherapy, Adjuvant ; statistics & numerical data ; Female ; Gastrectomy ; methods ; statistics & numerical data ; Humans ; Lymph Node Excision ; statistics & numerical data ; Lymphatic Metastasis ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Grading ; statistics & numerical data ; Neoplasm Invasiveness ; pathology ; Neoplasm Staging ; statistics & numerical data ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Stomach Neoplasms ; diagnosis ; mortality ; pathology ; therapy ; Survival Rate