Objective:To explore the clinicopathological features of serum alpha-fetoprotein-producing gastric cancer (AFPGC) and its subtype hepatoid adenocarcinoma of the stomach (HAS), and to analyze the related factors affecting the prognosis of AFPGC and HAS patients.Methods:The clinical data of 91 patients with AFPGC who were admitted to the Second Affiliated Hospital of Dalian Medical University from February, 2010 to February, 2021 were retrospectively collected. According to the results of hepatoid differentiation, the patients were divided into HAS group (26 cases) and non-HAS group (65 cases). Meanwhile, 130 patients with alpha-fetoprotein (AFP) negative gastric cancer at the Second Affiliated Hospital of Dalian Medical University were selected by stratified sampling method as common gastric cancer group. The clinicopathological data of patients in the HAS group, non-HAS group and common gastric cancer group were compared and all the patients were followed up for 3 years. Kaplan-Meier survival curves were plotted, and log-rank test was used to analyze the median survival time. Multivariate Cox regression was performed to analyze the independent risk factors affecting the prognosis of patients. Chi-square test was used for statistical comparison.Results:The proportion of patients with poorly differentiated tumor in the HAS group was higher than that in the non-HAS group (84.6%(22/26) vs. 55.4%(36/65)), and the difference was statistically significant ( χ2=7.02, P=0.030). The proportions of patients with age < 60 years old, abnormal level of carcinoembryonic antigen (CEA), vascular tumor thrombus, liver metastasis, poor differentiated tumor, and postoperative chemotherapy in the HAS group were higher than those in the common gastric cancer group (34.6% (9/26) vs. 13.8% (18/130), 26.9% (7/26) vs. 7.7% (10/130), 73.1% (19/26) vs. 51.5% (67/130), 30.8% (8/26) vs. 11.5% (15/130), 84.6% (22/26) vs. 37.7% (49/130), and 69.2% (18/26) vs. 27.7% (36/130), respectively), and the differences were all statistically significant ( χ2=5.16, 6.39, 4.06, 4.94, 18.73, and 16.52; all P<0.05). The proportion of male patients in the non-HAS group was lower than that in the common gastric cancer group (58.5%(38/65) vs. 73.1%(95/130)), while the proportions of patients with age < 60 years old, abnormal levels of carbohydrate antigen (CA)15-3, CA19-9, CA242, and CEA in the non-HAS group were higher than those in the common gastric cancer group (40.0% (26/65) vs. 13.8%(18/130), 7.7%(5/65) vs. 0, 23.1%(15/65) vs. 10.0%(13/130), 18.5%(12/65) vs. 7.7%(10/130), and 23.1%(15/65) vs. 7.7%(10/130), respectively), and the differences were all statistically significant ( χ2=4.27, 16.69, 11.25, 6.03, 5.02, and 9.18; all P<0.05). The proportion of patients with maximum diameter of tumor ≥ 5 cm, clinical stage Ⅲ or Ⅳ, lymph node metastasis, extrahepatic metastasis, and postoperative chemotherapy in the non-HAS group were all higher than those in the common gastric cancer group (58.5% (38/65) vs. 42.3% (55/130), 64.6% (42/65) vs. 46.2% (60/130), 83.1% (54/65) vs. 54.6% (71/130), 47.7% (31/65) vs. 15.4% (20/130), and 75.4% (49/65) vs. 27.7% (36/130), respectively), and the differences were all statistically significant ( χ2=4.53, 10.80, 15.26, 23.41, and 40.08; all P<0.05). The incidence of liver metastasis in patients with AFP >100 μg/L was higher than that in patients with AFP ≤100 μg/L (47.6%(10/21) vs. 17.1%(12/70)), and the difference was statistically significant ( χ2=8.18, P=0.004). The results of Kaplan-Meier survival analysis showed that the median survival time of patients in the HAS group, non-HAS group and common gastric cancer group was 13, 28, and 54 months, respectively, and the difference was statistically significant (log-rank test, χ2=20.33, P<0.001). The results of multivariate Cox regression analysis revealed that CA19-9 ( HR=5.803, 95% confidence interval (95% CI): 1.545 to 21.794, P<0.001) and extrahepatic metastasis ( HR=2.747, 95% CI: 1.243 to 6.068, P=0.012) were independent risk factors affecting the prognosis of patients in the non-HAS group. CA15-3 ( HR=84.163, 95% CI: 5.085 to 1 392.920, P=0.002), CA125 ( HR=0.038, 95% CI: 0.006 to 0.257, P=0.001), and the degree of tumor differentiation ( HR=2.284, 95% CI: 1.101 to 36.677, P=0.039) were independent risk factors affecting the prognosis of patients in the HAS group. Conclusions:Compared to common gastric cancer, AFPGC is characterized by advanced clinical stages, with higher propensity for lymph node metastasis, liver metastasis, extrahepatic metastasis and poor prognosis. The higher the AFP level before surgery, the more possibility of liver metastasis after surgery. Serum CA15-3 and CA125 might be tumor markers in predicting the prognosis of HAS patients.

Objective:To explore the clinicopathological features of serum alpha-fetoprotein-producing gastric cancer (AFPGC) and its subtype hepatoid adenocarcinoma of the stomach (HAS), and to analyze the related factors affecting the prognosis of AFPGC and HAS patients.Methods:The clinical data of 91 patients with AFPGC who were admitted to the Second Affiliated Hospital of Dalian Medical University from February, 2010 to February, 2021 were retrospectively collected. According to the results of hepatoid differentiation, the patients were divided into HAS group (26 cases) and non-HAS group (65 cases). Meanwhile, 130 patients with alpha-fetoprotein (AFP) negative gastric cancer at the Second Affiliated Hospital of Dalian Medical University were selected by stratified sampling method as common gastric cancer group. The clinicopathological data of patients in the HAS group, non-HAS group and common gastric cancer group were compared and all the patients were followed up for 3 years. Kaplan-Meier survival curves were plotted, and log-rank test was used to analyze the median survival time. Multivariate Cox regression was performed to analyze the independent risk factors affecting the prognosis of patients. Chi-square test was used for statistical comparison.Results:The proportion of patients with poorly differentiated tumor in the HAS group was higher than that in the non-HAS group (84.6%(22/26) vs. 55.4%(36/65)), and the difference was statistically significant ( χ2=7.02, P=0.030). The proportions of patients with age < 60 years old, abnormal level of carcinoembryonic antigen (CEA), vascular tumor thrombus, liver metastasis, poor differentiated tumor, and postoperative chemotherapy in the HAS group were higher than those in the common gastric cancer group (34.6% (9/26) vs. 13.8% (18/130), 26.9% (7/26) vs. 7.7% (10/130), 73.1% (19/26) vs. 51.5% (67/130), 30.8% (8/26) vs. 11.5% (15/130), 84.6% (22/26) vs. 37.7% (49/130), and 69.2% (18/26) vs. 27.7% (36/130), respectively), and the differences were all statistically significant ( χ2=5.16, 6.39, 4.06, 4.94, 18.73, and 16.52; all P<0.05). The proportion of male patients in the non-HAS group was lower than that in the common gastric cancer group (58.5%(38/65) vs. 73.1%(95/130)), while the proportions of patients with age < 60 years old, abnormal levels of carbohydrate antigen (CA)15-3, CA19-9, CA242, and CEA in the non-HAS group were higher than those in the common gastric cancer group (40.0% (26/65) vs. 13.8%(18/130), 7.7%(5/65) vs. 0, 23.1%(15/65) vs. 10.0%(13/130), 18.5%(12/65) vs. 7.7%(10/130), and 23.1%(15/65) vs. 7.7%(10/130), respectively), and the differences were all statistically significant ( χ2=4.27, 16.69, 11.25, 6.03, 5.02, and 9.18; all P<0.05). The proportion of patients with maximum diameter of tumor ≥ 5 cm, clinical stage Ⅲ or Ⅳ, lymph node metastasis, extrahepatic metastasis, and postoperative chemotherapy in the non-HAS group were all higher than those in the common gastric cancer group (58.5% (38/65) vs. 42.3% (55/130), 64.6% (42/65) vs. 46.2% (60/130), 83.1% (54/65) vs. 54.6% (71/130), 47.7% (31/65) vs. 15.4% (20/130), and 75.4% (49/65) vs. 27.7% (36/130), respectively), and the differences were all statistically significant ( χ2=4.53, 10.80, 15.26, 23.41, and 40.08; all P<0.05). The incidence of liver metastasis in patients with AFP >100 μg/L was higher than that in patients with AFP ≤100 μg/L (47.6%(10/21) vs. 17.1%(12/70)), and the difference was statistically significant ( χ2=8.18, P=0.004). The results of Kaplan-Meier survival analysis showed that the median survival time of patients in the HAS group, non-HAS group and common gastric cancer group was 13, 28, and 54 months, respectively, and the difference was statistically significant (log-rank test, χ2=20.33, P<0.001). The results of multivariate Cox regression analysis revealed that CA19-9 ( HR=5.803, 95% confidence interval (95% CI): 1.545 to 21.794, P<0.001) and extrahepatic metastasis ( HR=2.747, 95% CI: 1.243 to 6.068, P=0.012) were independent risk factors affecting the prognosis of patients in the non-HAS group. CA15-3 ( HR=84.163, 95% CI: 5.085 to 1 392.920, P=0.002), CA125 ( HR=0.038, 95% CI: 0.006 to 0.257, P=0.001), and the degree of tumor differentiation ( HR=2.284, 95% CI: 1.101 to 36.677, P=0.039) were independent risk factors affecting the prognosis of patients in the HAS group. Conclusions:Compared to common gastric cancer, AFPGC is characterized by advanced clinical stages, with higher propensity for lymph node metastasis, liver metastasis, extrahepatic metastasis and poor prognosis. The higher the AFP level before surgery, the more possibility of liver metastasis after surgery. Serum CA15-3 and CA125 might be tumor markers in predicting the prognosis of HAS patients.

Objective To construct the Hut78 cell line with EZH2 gene knocked into by CRISPR/Cas9 system. Methods The EZH2 expression vector pMD-18T-EZH2 with homologous arm and the sgRNA expression vector pSpCas9 (BB)-2A-Puro-sgRNA, which could cut the double stranded genomic DNA, were constructed, and the two vectors were co-transfected into Hut78 cells. Then the expression of EZH2 mRNA was detected by qPCR, and the expressions of EZH2 and H3K27me3 proteins were detected by Western blot assay. Results The pMD-18T-EZH2 and pSpCas9(BB)-2A-Puro-sgRNA recombinant vectors were confirmed by DNA sequencing. When Hut78 cells were transfected with the two recombinant plasmid, qPCR results showed that the expression of EZH2 mRNA was significantly increased, and Western blot analysis showed that the expressions of EZH2 and H3K27me3 proteins were significantly increased. Conclusion EZH2 gene is successfully knocked into Hut78 cells by CRISPR/Cas9 system.

Objective Cancer radio-therapy may induce bone damage of the patients.collagen type I gene expressions in osteoblast after radiation indicates the influence of radiation on the function of early and late osteoblast.Methods Bone marrow stromal cells were differentiated into osteoblasts in vitro.and the characteristics was indentified.The collagen type I expressions in early and late stage osteoblasts exposed to 1～4Gy radiation were examined by RT-PCR.Results Compared to control group,collagen type I gene expressions increased in early osteoblast after 1～3 Gy radiation (P＜0.05),while the gene expressions in late osteoblast that cultured 10 days decreased.Collagen type I gene expression in late stage ostoblast after 4 Gy irradiation was greatly higher than that in early stage osteoblast (P＜0.01).Conclusion After 1～3 Gy irradiation,the collagen type I expression in early osteoblast was enhanced,indicating the increased ability of bone formation.The exposure to 1～3 Gy decreased collagen type I expression in late osteoblast and weakened the ability of bone formation.The result of high expression of collagen type I in late osteoblast after 4 Gy irradiation may be the manifestation of compensatory function.