Objectvi e To investigate whether promoting gap junctions may contribute to the radiosensi-tivity in triple negative breast cancer( TNBC)cells.Methods HCC70(triple-negative),MCF-7(ER-posi-tive)or SK-BR3(HER2-positive )cells were transfected with pcDNA/5 -Cx43 expression plasmid using liposome 2000.The transfected cells were treated with various doses of radiation(0,5,10,15 Gy),and the level of Cx43 protein was determined by Western blot and the cell connectivity was determined by fluorescent tracer technique. Cell proliferation inhibition,clone formation ability and apoptosis were detected using MTT,clone formation assay, AnnexinV-FITC/PI double staining and flow cytometer,respectively.Results The level of Cx43 protein signifi-cantly increased in HCC 70 -Cx43 ,MCF-7 -Cx43 and SK-BR3 -Cx43 cells.After transfection the cells were treated with various doses of radiation,level of Cx43protein was gradually enhanced in dose dependent fashion .The re-sults form fluorescent tracer technique showed that fluorescence intensity was gradually elevated with increase of radiation doses.Cell viability and clone formation ability were decreased gradually in dose dependent manner in HCC70-Cx43 ,MCF-7 -Cx43 and SK-BR3-Cx 43 cells.Unexpectedly,the inhibitive effect of proliferation ability and clone formation ability in HCC70 -Cx43 cell was higher than in MCF-7 -Cx43 and SK-BR3 -Cx43 cells under same conditions.The results from AnnexinV-FITC/PI and flow cytometer showed that apoptosis rate was enhanced gradually accompanying with increase of radiation doses.Conclu sion Enhancing the function of cell gap junc-tions promoted radiosensitivity of breast cancer cells,particularly in TNBC cells.Radiation can strengthen cell gap junctions in breast cancer cell and cytotoxicity of TNBC cell can be enhanced by both synergistic effects.

In the present study, the fluoro-gold(FG) and horseradish peroxidase(HRP) combined tracing method was used to investigate the localization of parasympathetic preganglionic neurons and ascending projection neurons in lumbosacral spinal cord of the rat. FG was injected into the lateral parabrachial nucleus(PBL) or into Barrington's nucleus on one side, and HRP was applied to the contralateral pelvic nerve. The retrogradely FG-labeled neurons were found in bilateral "visceral field" at segments L_5-S_2, and the majority of them were concentrated in the intermediolateral nucleus (IML), and the dorsal commissural nucleus (DCN). In addition to these areas, some labeled neurons were also observed in bilateral lamina I and lateral spinal nucleus (LSN). The parasympathetic preganglionic neurons labeled with HRP were seen in the IML at segments L_6-S_1, occasionally appeared in the intercalated nucleus. In the IML area, HRP-labeled parasympathetic preganglionic neurons were located in its ventral part, however, the localization of FG-labeled neurons projected to the PBL and Barrington's nucleus were mainly found in the dorsal and dorsomedial part of the IML, and a few FG-labeled cells were scattered among HRP-labeled cells. Based on the present and other investigations, the nomenclature, organization and function of the IML and the composition of the LSN were discussed.

Objective To examine the localization of neurokinin B receptor (NK3)\|like immunoreactivity (\|LI) in the central nervous system of the mouse. Methods An immunohistochemcial staining method was used. Results NK3 receptor\|LI was localized in somatic and dendritic profiles in the most parts and in neuropil in a few regions of the mouse central nervous system. A large number of neurons with NK3\|LI was seen in the anterior olfactory nuclei, accumbens nucleus, septal area, ventral pallidum, pallidum, caudate putamen, nucleus of the stria terminalis, anterior hypothalamic area, tuber cincreum area, lateral hypothalamic area, perifornical nucleus, supraoptic nucleus, arcuate nucleus, mammillar nuclei, substatia nigra, ventral tegmental area, retrorubral area, superior and inferior colliculus, periaqueductal gray, nucleus of the solitary tract, and superficial layers of the medullary and spinal dorsal horns. The superfical layers of the cerebral cortex, piriform cortex, dorsal hippocampus, amygdal complex, reticular formation of the brainstem contained some neurons with NK3 receptor\|LI. In the ventral hippocampus, median and intralaminar nuclei of the thalamus and interpeduncular nuclei, NKR\|LI was localized in neuropil. Conclusion\ Neurons with NK3 receptor\|LI were widely distributed in the central nervous system. It may be involved in many physiological functions in the central nervous system of the mouse.\;[

Objective This study is to explore the differences in the curative effect of intra -abdominal infusion between Recombinant mutant human tumor necrosis factor and Dendritic cell -Cytokine induced killer . Methods We selected 48 advanced gastric cancer patients with ascites .Those patients were randomized into two groups:DC-CIK group and rmhTNF group .After one month treatment ,we observed the adverse events both two groups,and evaluated the clinical beneficial responses ,the responsive rate ,the tumor indicators′level,the immune indexes and the time of tumor progression .Results In the DC-CIK group,the RR and the CBR were 83.3%and 66.67%,respectively;while in the rmhTNF group,they were 58.33%and 75%,respectively.The difference between two groups was statistically significant (P<0.05).Only CA724 decreased after treatment in two groups (P=0.015).There were no significant differences of tumor markers between the two groups before and after treatment(P>0.05).The ratios of CD3 +、CD4 +、NK cells distinctly increased after treatment (P<0.05),but the ratio of CD8 +cell was no obvious difference in DC -CIK group(P=0.551);The ratio of NK cells increased obviously after treatment(P=0.027),but the ratios of CD3 +and CD4+were similar as previous in rmhTNF group(P>0.05).One year follow-up of the time to progression(TTP)was 7.1 months in DC-CIK group,and 5.8 months in TNF group,which was statistically significant (P=0.02).Conclusion Both rmhTNF and DC-CIK can improve the efficacy for the patients with malignant ascites caused by gastric cancer .However ,DC-CIK immunotherapy shows better active effect on improvement of specific immune function .

FOS protein in the lumbosacral spinal cord of the rat was detected to study neuronal activation induced by noxious stimulation of the urethra. In rats receiving infusion of 100 μl of 2% formalin into the urethra, a large number of FOS-positive neurons were seen in the lumbosacral cord segments (L6 and S1), and they were primarily distributed in the medial dorsal horn and dorsal commissural nucleus. Nearly all of FOS expression was blocked by bilateral transection of the pudendal nerve, whereas bilateral transection of the pelvic nerve seemed to have no obvious effect on FOS expression.Behavioral changes were also observed in the nerve-transected rats. Bilateral pelvic nerve-transected rats showed frequently licking of the external urethral orifice after application of the irritant to the urethra, which was quite similar to that of the nerve-intact rats. Such behavior completely disappeared in the rats receiving bilateral pudendal nerve transection. The results suggest that the medial dorsal horn and dorsal commissural nucleus in the L6 and S1 segments are involved in processing nociceptive inputs from the urethra, which is carried by the pudendal nerve.

Objective: This study aimed to compare and analyze the functional differences between peripheral blood mono-cyte-derived dendritic cells (DCs) of Helicobacter pylori-positive and H. pylori-negative patients with gastric cancer. Methods:H. py-lori infection was detected in 84 patients with gastric cancer in our hospital from January 2011 to October 2012 by the 14C-urea breath test. DCs were generated from monocytes isolated by an adherent method from the two groups of patients and cultured in the presence of rhIL-4, rhGM-CSF, and rhTNF-α. Furthermore, the expression of surface marker molecules was determined by fluorescence-activat-ed cell sorting analysis. The cytotoxicity of DCs pulsed T cells against gastric carcinoma cell was assessed by the lactate dehydroge-nase-releasing assay. The secretion of IL-12 and IFN-γin the supernatant was determined by enzyme-linked immunosorbent assay. Re-sults:No difference was observed in the morphological change of the maturation process. The mean expression of CD1a, CD80, CD83, CD86, and HLA-DR molecules in DCs of H. pylori-infected patients was higher than that in DCs of H. pylori-negative group, and the differences were statistically significant except for CD1a and HLA-DR. The cytotoxicity activities, IL-12 release, and IFN-γrelease in the H. pylori-positive group were significantly higher than those in the H. pylori-negative group (P<0.05). Conclusion:H. pylori infec-tion has no effect on the morphological change of the maturation process of monocyte-derived DCs. These data clearly demonstrate that monocyte-derived DCs of H. pylori-infected patients with gastric cancer can induce stronger maturation and activation than those of H. pylori-negative patients.

Objective To investigate the endoscopic prevalence of erosive esophagitis (EE) among 13 hospitals in Guangdong province of China. Methods Retrospectively reviewed all the cases (63459 cases) that received oesophagogastrodeuodenoscopy in 13 main hospitals in Guangdong province of China in 2003. Los Angeles criteria for classification of erosive esophagitis were employed as the basis of analysis. Results One thousand two hundreds and sixty-three patients (age range 3-90yr, mean 50. 2 ?17. 1 ) were found to have EE. The overall prevalence of EE was 1. 99% (1263/63459). The prevalence of EE in A, B, C, and D grade were 0. 94% , 0. 69% , 0. 21% and 0. 14% respectively. Age correlated positively on endoscopic grading of EE (F=22. 932, P

Toll-like receptor 5 (TLR5) is responsible for the recognition of bacterial flagellin in vertebrates. In the present study, the first TLR5 gene in duck was cloned. The open reading frame (ORF) of duck TLR5 (dTLR5) cDNA is 2580 bp and encodes a polypeptide of 859 amino acids. We also cloned partial sequences of myeloid differentiation factor 88, 2'-5'-oligoadenylate synthetase (OAS), and myxovirus resistance (Mx) genes from duck. dTLR5 mRNA was highly expressed in the bursa of Fabricius, spleen, trachea, lung, jejunum, rectum, and skin; moderately expressed in the muscular and glandular tissues, duodenum, ileum, caecum, and pancreas; and minimally expressed in the heart, liver, kidney, and muscle. DF-1 or HeLa cells transfected with DNA constructs encoding dTLR5 can activate NF-kappaB leading to the activation of interleukin-6 (IL-6) promoter. When we challenged ducks with a Herts33 Newcastle disease virus (NDV), mRNA transcription of the antiviral molecules Mx, Double stranded RNA activated protein kinase (PKR), and OAS was up-regulated in the liver, lung, and spleen 1 and 2 days post-inoculation.
2',5'-Oligoadenylate Synthetase/genetics/metabolism ; Animals ; Cell Line ; *Cloning, Molecular ; Ducks ; Gene Expression Regulation/*physiology ; Humans ; Immunity, Innate ; Myeloid Differentiation Factor 88/genetics/metabolism ; Myxovirus Resistance Proteins/genetics/metabolism ; Newcastle Disease/metabolism ; Newcastle disease virus/classification ; RNA, Messenger/genetics/metabolism ; Species Specificity ; Toll-Like Receptor 5/genetics/*metabolism

Objective To observe the effect of anti NRP-1 b1/b2 monoclonal antibody (NRP-1mAb) on migration and invasion of gastric cancer cell line BGC-823, and to explore the possible mechanism. Methods NRP-1mAb was prepared in the laboratory, and the purity of antibody was detected by flow cytometry. The different concentrations of NRP-1mAb were added into the culture medium of gastric cancer cell line BGC-823. The migration and invasion of cells after 12 hours was observed by using Transwell method. The phosphorylation of related signal proteins after NRP-1mAb was detected by Western blot analysis. Results When NRP-1mAb prepared by patented technology had the effects on BGC-823 cells after 12 hours, the number of migration and invasion of BGC-823 cells was reduced. The number of cells through the basement membrane in the control group (blank) and the administration group (NRP-1mAb 25, 100, 400 μg / ml) were 167 ± 9, 138 ± 5, 98 ± 5, 36 ± 4, respectively (F = 22.6, P< 0.01); the number of cells through the filtration membrane were 231 ± 40,224 ± 19,176 ± 26,124 ± 34,respectively(F=26.63,P<0.01). There were statistically significant differences between the administered group and the control group at 100 and 400 μg/ml (all P< 0.001). High concentration of NRP-1mAb (100 μg/ml) decreased the phosphorylation level of Akt after 10 minutes' function on gastric cancer cells. However, it was difficult to detect phosphorylated Akt after 30 minutes. Conclusion NRP-1mAb may inhibit the migration and invasion of gastric cancer cell line BGC-823 by decreasing the phosphorylation of Akt, which is positively correlated with the concentration.

Objective:To explore the efficacy of adjusting the dose of imatinib dose in the context of therapeutic drug monitoring (TDM) in patients with gastrointestinal stromal tumors (GISTs) who are receiving adjuvant therapy after complete resection of their tumors.Methods:This was a descriptive study. Inclusion criteria were (1) complete surgical resection with a pathological diagnosis of GIST, (2) postoperative adjuvant therapy with imatinib and dosage adjustment, (3) multiple TDM of imatinib, and (4) complete clinical, pathological, and follow-up data. The data of 70 patients with GISTs treated at Union Hospital, Tongji Medical College, Huazhong University of Science and Technology between January 2015 and December 2023 were collected retrospectively. The study cohort comprised 15 (21.4%) men and 55 (78.6%) women of median age 60 years (range: 25–82). Of the eligible patients, 49 (70.0%) were at high-risk, 14 (20.0%) at intermediate-risk, six (8.6%) at low-risk, and one (1.4%) at very low risk. Patients were followed up by the gastrointestinal stromal tumor clinic every 2–3 months and their plasma concentrations of imatinib were checked. The dose was adjusted to 300 mg/d or 200 mg/d depending on whether they had had ≥ grade III adverse reactions, and whether the first plasma concentration of imatinib was ≥ 1,500 μg/L or between the expected range of 760 μg/L–1,100 μg/L. Studied indicators included adverse reactions, quality of life before and after dose adjustment, and overall survival and recurrence-free survival (RFS) after dose adjustment.Results:Before dose adjustment, all 70 patients received 400 mg of imatinib daily, with initial TDM values of 1,900 ± 568 μg/L, for a median duration of 8.3 months. After dose adjustment, 60 patients received 300 mg daily, with a TDM of 1,216 ± 350 μg/L, whereas 10 received 200 mg daily, with a TDM of 1,023 ± 269 μg/L. The median duration of treatment after dose adjustment was 23.4 months. Compared with those whose dosages were not adjusted, the incidence of bone marrow suppression was significantly lower (74.3% [52/70] vs. 51.4% [36/70], χ 2=9.202, P=0.010); as were the incidences of edema (95.7% [67/70] vs. 50.0% [35/70], χ 2=40.526, P<0.001); skin reactions (70.0% [49/70] vs. 32.9% [23/70), χ 2=22.495, P<0.001); and gastrointestinal reactions (38.6% [27/70] vs. 10.0% [7/70], χ 2=15.899, P<0.001) in those whose dosages were adjusted. The average total scores for physical health before and after dose adjustment were 76 ± 5 and 88 ± 4, respectively; whereas the mental health scores were 75 ± 6 and 89 ± 4, respectively. The median follow-up period was 36 months (range 6–126). During the first 3 years of follow-up, five high-risk patients with non-gastric GISTs developed recurrences. The 3-year overall survival rate was 100%, and the 3-year RFS rate was 92.8%, high-risk patients having a 3-year RFS rate of 89.8%. Conclusion:The adverse reactions and quality of life of GIST patients with severe adverse reactions to adjuvant imatinib therapy after complete resection can be mitigated by appropriately reducing the dosage of imatinib under the guidance of TDM.