ObjectiveTo dynamically observe and evaluate the damage to the pulmonary air-blood barrier in mice during the inflammatory cancer transformation process of ulcerative colitis （UC） based on the "lung-intestine correlation" theory. MethodsSixty-five C57BL/6 mice were divided into a normal group （n=25） and a model group （n=40） using a random number table. Azoxymethane/dextran sodium sulfate （DSS） method was used to establish a mouse model of UC inflammation cancer transformation in the modeling group. According to the tissue collection time points at 5， 8， 11， 13， and 15 weeks， the normal group mice were randomly divided into the normal 5w， 8w， 11w， 13w， and 15w groups. The model group mice， 10 mice of which died after the first cycle of DSS administration， were randomly divided into model 5w， 8w， 11w， 13w， and 15w groups. During the experiment， the general condition of the mice was observed daily， and their body weight was measured weekly. At the corresponding tissue collection time points， the colon length of each group was measured. Histopathology of mouse lung and colon tissues was examined using HE staining. Immunofluorescence was used to detect changes in the positive expression of tight junction protein （ZO-1）， vascular endothelial cadherin （VE-cadherin）， and cytoskeletal protein （F-actin） in lung and colon tissues. RT-PCR was used to detect the mRNA expression of apoptosis regulatory proteins B-cell lymphoma-2 （Bcl-2）， BCL2-associated X protein （Bax）， and Cysteine aspartic acid protease-3 （Caspase-3） in lung tissues. Western Blot was employed to measure protein levels of ZO-1， VE-cadherin， and F-actin in lung tissues. ResultsCompared to the normal group at the same time point， the mice in the model group at each time point generally had poorer conditions， with weight loss and shortened colon length （P＜0.05 or P＜0.01）. In the model 5w group， there was significant inflammatory cell infiltration in the colon tissue； in the model 8w group， there was mild atypical hyperplasia； in the model 11w group， the crypt structure was disordered， and moderate to severe atypical hyperplasia occurred； in the model 13w and 15w groups， tumors appeared. Pulmonary interstitial lesions， inflammation， vasculitis， and fibrosis were observed at all stages of UC inflammation cancer transformation. The protein levels of ZO-1， VE-cadherin， and F-actin， as well as Bcl-2 mRNA expression in lung tissue decreased during the acute inflammatory recovery period， atypical hyperplasia period， and canceration period， while the expressions of Bax and Caspase-3 mRNA increased； the expressions of ZO-1， VE-cadherin， and F-actin proteins in colon tissue decreased during the acute inflammatory recovery period， atypical hyperplasia period， and canceration period （P＜0.01 or P＜0.05）. Compared to the model 5w group， the ZO-1 and F-actin protein levels and Bcl-2 mRNA expression in lung tissue in the other model groups increased in the atypical hyperplasia period and canceration period， while the expressions of Bax and Caspase-3 mRNA decreased； the expression of ZO-1 protein in colon tissue increased in the canceration period， and the expression of VE-cadherin protein decreased in the atypical hyperplasia period （P＜0.01 or P＜0.05）. ConclusionIn the process of "inflammatory response-atypical hyperplasia-carcinogenesis" in UC inflammatory cancer transformation mice， there were damage to air-blood barrier.

Objective:To explore the mechanism of modified Shaoyao Decoction with Astragali Radix and Houttuynize Herba in regulating PI3K/Akt/NF-κB pathway against intestinal mucosal barrier injury in ulcerative colitis (UC) rats.Methods:Totally 72 Wistar rats were randomly divided into a blank control group, a model group, a mesalazine group, and modified Shaoyao Decoction high-, medium-, and low-dose groups using a random number table method. After successful modeling, the mesalazine group was given 0.2 g/kg of mesalazine suspension by gavage, while modified Shaoyao Decoction high-, medium-, and low-dose groups were given 44.5, 22.3, and 11.1 g/kg of modified Shaoyao Decoction by gavage, once a day, for a total of 4 weeks. The pathological changes of colon tissue were observed using HE staining; immunofluorescence assay was used to detect the expressions of tight junction protein (ZO-1), cytoskeletal protein (F-actin), and vascular endothelial cadherin (VE cadherin) in colon tissue; Western blot was used to detect the protein expressions of PI3K, Akt, NF-κB, VEGF, and cyclooxygenase (COX-2) in colon tissue; QRT PCR was used to detect the expressions of PI3K and Akt mRNA in colon tissue; ELISA method was used to detect the levels of serum TNF-α and endothelin (ET).Results:Compared with the model group, the DAI scores of the mesalazine group and modified Shaoyao Decoction high-dosage group decreased significantly on the 7th, 14th, 21st and 28th day after modeling ( P<0.05 or P<0.01). In mesalazine group and modified Shaoyao Decoction high-, medium-, and low-dose groups, the pathological injury score of colon tissue decreased ( P<0.01), the immunofluorescence intensity of ZO-1, F-actin and VE-cadherin protein expression in colon tissue increased ( P<0.05 or P<0.01), and the expressions of PI3K, Akt, NF-κB and VEGF protein decreased ( P<0.01). The expression of COX-2 protein and the levels of PI3K mRNA and Akt mRNA decreased in the mesalazine group and modified Shaoyao Decoction high- and medium groups ( P<0.05 or P<0.01). Conclusion:Modified Shaoyao Decoction can antagonize the intestinal barrier injury of UC by inhibiting PI3K/Akt/NF-κB pathway and promote the healing of intestinal mucosal ulcer.

Objective To investigate the effect of serum of esophageal squamous cell carcinoma (ESCC) patients with blood stasis syndrome (BSS) on proliferation and cycle of EC9706 cells, and to explore the action of blood micro-environment of ESCC patient with BSS on EC9706 cells. Methods Human EC9706 cells were cultured in an incubator with RPMI-1640 medium containing fetal bovine serum ( FBS) , at 37℃ and under 5% saturated humidity for 24 h. After EC9706 cells were starved in serum-free medium for another 24h, the three experimental groups were treated with serum of ESCC patients with BSS, serum of ESCC patients with spleen-qi deficiency syndrome (SQDS), and serum from healthy volunteers, respectively. Cell proliferation was determined by methyl thiazolyl tetrazolium ( MTT) assay, EC9706 cell morphology was observed under light microscope, and cell cycle was measured by flow cytometer (FCM). Results The serum concentrations of ESCC patients with BSS and ESCC patients with SQDS for obtaining 50 percent cell proliferation rates ( PI50) were 71.1 μL/mL and 118 μL/mL, respectively. And the proliferation of EC9706 cells in the both groups all arrived to the peak values at culturing hour 48. The light microscopy results showed that the feature of EC9706 cells in both groups presented as spindle-like or polygon-like shape, and cell count in BSS group was larger than SQDS group. FCM assay results for EC9706 cell cycle showed that the percentage of G1-phase EC9706 was decreased and the percentage of S-phase EC9706 was increased in BSS group as compared with those in SQDS group ( P<0.05). Conclusion Serum micro -environment in ESCC patients with BSS is more beneficial to EC9706 cells proliferation than ESCC patients with SQDS, and the mechanism may be related to the regulation of cell cycle.

Purpose To explore the role of tumor necrosis factor-α( TNF-α) in the process of temozolomide ( TMZ) reduce glioma in-vasiveness and its possible mechanism. Methods C6 glioma cells of logarithmic phase were randomly divided into TMZ treatment (10, 30, 60, 120, 180, 240 min group) (n=15), dynamic monitoring content of TNF-αin the culture medium was measured by ra-dioimmunoassay, expression of p53 protein in C6 cells was detected with Western blotting method, and cell apoptosis was used with AnnexinV-FITC. A glioma invasiveness model was established in vitro and glioma invasiveness was determined by crystal violet stai-ning. Results For C6 cells, contents of TNF-αin the nutrient fluid and expressions of p53 protein in C6 cells obviously increased af-ter TMZ treatment and they achieved the peak at 120 min (P<0. 01), followed by decrease gradually. Glioma invasiveness was re-duced after TMZ acted on glioma in vitro. Conclusion TMZ can reduce glioma invasiveness by TNF-α, which this role may be is TMZ promote C6 cells release of TNF-α and increased TNF-α due to glioma cells apoptosis.