Objective To determine the effect of cetuximab(C225)on the radioresistant human esophageal squamous carcinoma eell line KYSE-150R.Methods A radioresistant human esophageal squamous carcinoma cell line KYSE-150R was established by fractionated irradiation.Morphological changes from KYSE-150 to KYSE-150R were observed by phase-contrast microscopy.Karyotype analysis was performed by G-banding.The radiosensitivities were analyzed by colony formation assays.Results The population doubling time of KYSE-150 and KYSE-150R were(23.6±0.2)h and(25.9±0.6)h (t=6.6,P<0.01),respectively.The chromosome number of KYSE-150R was increased and chromosome aberrations were observed from(69.3±1.9)h to(73.7±1.2)h(t=-8.83,P<0.01).The SF2,D0,Dq and N values of KYSE-150R were all higher than those of KYSE-150.After 5μg/ml of C225 added,the SF2,D0,Dq and N values were significantly decreased as compared to the control.After C225 treatment,the G0/G1 and G2/M phase cells were increased,while S-phase cells decreased(t=-4.478-4.308,P<0.05).Conclusion Cetuximab can enhance the radiosensitivity of radioresistant human esophageal squamous carcinoma cell line KYSE-150R.

Objective To investigate the potential mechanism of intestinal trefoil factor(ITF)against methotrexate (MTX)- induced injury in intestinal mucosa. Methods Cultured IEC-6 cells were divided into groups as follows: blank group, MTX treated group, ITF treated group and experimental group treated with gradient concentrations of ITF plus MTX. Expression of E-cadherin mRNA was determined by Real-Time polymerase chain reaciton (RT- PCR). The activity of matrix metalloproteinase(MMP)-2 and MMP-9 was measured by gelatin zymogramphy. Caspases-3 activity was measured by colorimetric assay. Cell proliferation was assessed by cell counting kit-8 (CCK-8)assay. Migration of IEC-6 in vitro was observed using modified Boyden chamber assay. Results The expression of E-cadherin mRNA in experimental group (treated with 0.1 mg/ml or 1 mg/ml of ITF) was significantly down-regulated (0. 538±0. 109 or 0. 528±0. 132, respectively) in comparison with MTX treated group (0. 763±0. 139) with significant difference (P=0. 021 or P=0. 025, respectively). There was no significant difference in activity of MMP-2 and MMP-9 among groups (P＞0. 05). When compared with MTX treated group (0. 090 ±0. 011 ), the activity of Caspase3 in experimental group (treated with 0. 1 mg/ml or 1 mg/ml of ITF) was significantly decreased (0. 077±0. 009, P=0. 032 or 0. 044±0. 009,P=0. 005, respectively). There was no statistical difference in cell proliferation between experimental group (treated with 1 μg/ml, 0.01 mg/ml, 0. 1 mg/ml or 1.0 mg/ml of ITF) and MTX treated group (P=0. 132,0. 150,0. 114 or 0. 367, respectivley). More migratory cells attached to the bottom surface of the membrane in experiment group (treated with 0. 1 mg/ml or 1 mg/ml of ITF) in comparison with MTX treated group (P ＜0. 001 ). Moreover, more migratory cells were found in experimental group treated with 1.0 mg/ml of ITF than those in group treated with 0. 1 mg/ml of ITF (P＜0. 001). Conclusions Without cell proliferation, the protective effect of ITF is related to its functions of promoting cell migration and inhibiting cell apoptosis, which may down-regulate expression of E-cadherin mRNA.

Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.