Background and purpose:Differentiation of tumor tissue is an important factor on determining the prognosis of gastric cancer. This study aimed to investigate the expression levels and clinical signiifcance of gender determining region Y-box 2 (SOX2) gene and octamer binding factor 4 (OCT4) gene in gastric cancer tissues varying different differentiation degrees. Methods: Sixty cases with gastric cancer were recruited in this study. The gastric cancer tissues and corresponding normal mucosa of the 60 cases were obtained. The mRNA and protein level of SOX2, OCT4 gene are evaluated by the quantitative real-time PCR (qRT-PCR), Western blot and immunohistochemistry, respectively. The relationship between the expression levels of SOX2, OCT4 gene and clinical pathological parameters were also analyzed in this study. Results:The expression of SOX2 in both mRNA and protein levels had no signiifcant difference between the well-differentiated gastric cancer tissues and normal gastric mucosa (mRNA levels:t=0.1033, P>0.05;protein levels:t=0.116, P>0.05). However, both the mRNA and protein expression of SOX2 in patients with well-differentiated gastric cancer tissues were signiifcant higher than not only in patients with moderately differentiated gastric carcinoma (mRNA levels: t=12.48, P<0.05; protein levels: t=22.78, P<0.05) but also in patients with than poorly differentiated gastric carcinoma (mRNA levels:t=17.56, P<0.05;protein levels:t=30.00, P<0.05). In contrast to SOX2, both the mRNA and protein expression of OCT4 in patients with well-differentiated gastric cancer tissues were signiifcant lower than not only in patients with moderately differentiated gastric carcinoma (mRNA levels:t=13.23, P<0.05; protein levels: t=25.56, P<0.05) but also in patients with poorly differentiated gastric carcinoma (mRNA levels: t=12.10, P<0.05; protein levels: t=69.48, P<0.05). There was no significance of OCT4 mRNA and protein expression between the well-differentiated gastric cancer tissues and normal gastric mucosa (mRNA levels:t=2.436, P>0.05;protein levels:t=1.064, P>0.05). Immunohistochemical study demonstrated that the positive rate of SOX2 in patients with well-differentiated gastric cancer tissues (10/21) were higher than in patients with not only moderately differentiated gastric carcinoma (7/20) but also poorly differentiated gastric carcinoma (2/19, P<0.05), while the positive rate of OCT4 in cases with well-differentiated gastric cancer tissues (2/21) were lower than in cases with not only moderately differentiated gastric carcinoma (6/20) but also the poorly differentiated gastric carcinoma (12/19, P<0.05). There was no correlation between the expression of SOX2, OCT4 in gastric cancer and gender or age (P>0.05). Nevertheless, the expression of SOX2, OCT4 were positive or negative correlated with the pathological staging, the degree of inifltration and lymph node metastasis (P<0.05). Conclusion:Decreased SOX2 expression and increased expression level of OCT4 can promote the formation, development and invasion of gastric cancer and they may become biomarkers or the diagnosis, treatment and prognosis evaluation in gastric carcinoma.

Objective To study on the procedure, safety and effectiveness of polyetheretherketone (PEEK) implant applied to cranioplasty of skull defect. Methods A total of 11 cases (10 male, 1 female) of unilateral skull defect, more than 6 months post operation, were included in this study. PEEK implant was custom-made by three-dimensional numerically controlled processing depended on the data obtained from 1 mm-slice CT scan before cranioplasty individually. Autoclaved implants were applied to cranioplastic surgeries under general anesthesia. Findings of imaging examination and vital signs were compared before and after operation. Vital sign changes and circumstances during procedure were noted, and following-up reviews were performed on 2-week, 3-month and 1 year after operation respectively. Results Wounds healing were uneventful in 11 cases, no postoperative paresthesia presented. Surgical complications including temporary subcutaneous exudates were cleared up by puncture and aspiration in 5 cases, subcutaneous hematoma duo to drainage removal in 1 case, of which an emergency evacuation was performed and the implant was still with instant incision closure and then smooth recovery eventually. No significant abnormal fluctuation of laboratory examination was reported, moreover no artifact interference was found on postoperative CT scan or MR image. The subjective feelings and external sensory effects are satisfactory in patients. No emerging dysfunction of central nervous system or other organs was found, and no long-term complication was appeared. Conclusion It is feasible and safe to apply PEEK implant to cranioplasty without additional operative difficulty. This kind of material is an ideal alternative for repairing skull defect to patients with good financial condition and specific demands for postoperative status especially.

Objce tive To investigate the effect and significance of down-regulation of Oct4 gene on biological characteristics of MDA-MB-231 breast cancer stem cells.Methods Breast cancer cell line MDA-MB-231 cells were used in this study.Breast cancer stem cells were isolated and enriched by serum-free culture.The obtained stem cells were identified through calculating the percentages of CD44 and CD24 stem cells by FACS and evaluating the paclitaxel resistance in vitro and tumorigenicity in mice.RT-PCR, real-time PCR (qPCR) and Western blot were used to detect Oct4 expression.RNA interference was applied to induce Oct4 down-regulation.The interference experiment set up a control group ( no siRNA transfection) , negative control group ( negative siRNA group,transfection of siRNA sequences without any interfering effect on the cells) and Oct4 siRNA group ( transfection of siRNA with interfering effect on the Oct4 gene) .Methyl thiazolyl tetrazolium ( MTT ) and Transwell chamber tests were conducted to detect the proliferation and invasion ability of MDA-MB-231 breast cancer stem cells after Oct4 knock-down, and paclitaxel inhibition test was applied to evaluate drug resistance of MDA-MB-231 breast cancer stem cells after Oct4 knock-down. Resulst MDA-MB-231 breast cancer stem cells grew as spheres cultured in serum-free suspension.MDA-MB-231 breast cancer stem cells showed a higher percentage of CD44+C/D24 -/low cells (97.2%) than that in MDA-MB-231 breast cancer cells ( 76.6%) ( P<0.05) .The tumor size in mice inoculated with MDA-MB-231 breast cancer stem cells was (124.60±13.65)mm3, significantly larger than that of mice inoculated with breast cancer cells (68.20±9.99 mm3) (P=0.0007).MDA-MB-231 breast cancer stem cells were less sensitive to paclitaxel inhibition than MDA-MB-231 breast cancer cells showing by 50% inhibitory concentration (IC50) [(4.40±0.48) μg/ml vs.(8.20±0.34) μg/ml, P<0.05].However, the expression of transcriptional factors Oct4 was higher in MDA-MB-231 breast cancer stem cells than that in breast cancer cells (P<0.05).The proliferation potential of MDA-MB-231 breast cancer stem cells with Oct4 siRNA interference was significantly lower than that in the negative siRNA and control groups ( P<0.05) from the third day.The invasion ability of MDA-MB-231 breast cancer stem cells with Oct4 siRNA interference was obviously reduced than that in the control and negative siRNA groups shown by number of penetrated cells [(46.52±2.58) vs.(79.67±3.85) and (77.29±2.13), P<0.05 for both].As for resistance to paclitaxel, IC50 of MDA-MB-231 breast cancer stem cells with Oct4siRNA interference was significantly decreased [(4.48±0.22) μg/ml] compared with that in the control [(7.99±0.59) μg/ml] and negative siRNA group [(8.10±0.68) μg/ml] (P<0.05 for both).Conclusions MDA-MB-231 breast cancer cells are successfully obtained by serum-free culture. The proliferation potential, invasion ability and drug resistance of breast cancer stem cells were down-regulated by Oct4 gene knock-down.

Objce tive To investigate the effect and significance of down-regulation of Oct4 gene on biological characteristics of MDA-MB-231 breast cancer stem cells.Methods Breast cancer cell line MDA-MB-231 cells were used in this study.Breast cancer stem cells were isolated and enriched by serum-free culture.The obtained stem cells were identified through calculating the percentages of CD44 and CD24 stem cells by FACS and evaluating the paclitaxel resistance in vitro and tumorigenicity in mice.RT-PCR, real-time PCR (qPCR) and Western blot were used to detect Oct4 expression.RNA interference was applied to induce Oct4 down-regulation.The interference experiment set up a control group ( no siRNA transfection) , negative control group ( negative siRNA group,transfection of siRNA sequences without any interfering effect on the cells) and Oct4 siRNA group ( transfection of siRNA with interfering effect on the Oct4 gene) .Methyl thiazolyl tetrazolium ( MTT ) and Transwell chamber tests were conducted to detect the proliferation and invasion ability of MDA-MB-231 breast cancer stem cells after Oct4 knock-down, and paclitaxel inhibition test was applied to evaluate drug resistance of MDA-MB-231 breast cancer stem cells after Oct4 knock-down. Resulst MDA-MB-231 breast cancer stem cells grew as spheres cultured in serum-free suspension.MDA-MB-231 breast cancer stem cells showed a higher percentage of CD44+C/D24 -/low cells (97.2%) than that in MDA-MB-231 breast cancer cells ( 76.6%) ( P<0.05) .The tumor size in mice inoculated with MDA-MB-231 breast cancer stem cells was (124.60±13.65)mm3, significantly larger than that of mice inoculated with breast cancer cells (68.20±9.99 mm3) (P=0.0007).MDA-MB-231 breast cancer stem cells were less sensitive to paclitaxel inhibition than MDA-MB-231 breast cancer cells showing by 50% inhibitory concentration (IC50) [(4.40±0.48) μg/ml vs.(8.20±0.34) μg/ml, P<0.05].However, the expression of transcriptional factors Oct4 was higher in MDA-MB-231 breast cancer stem cells than that in breast cancer cells (P<0.05).The proliferation potential of MDA-MB-231 breast cancer stem cells with Oct4 siRNA interference was significantly lower than that in the negative siRNA and control groups ( P<0.05) from the third day.The invasion ability of MDA-MB-231 breast cancer stem cells with Oct4 siRNA interference was obviously reduced than that in the control and negative siRNA groups shown by number of penetrated cells [(46.52±2.58) vs.(79.67±3.85) and (77.29±2.13), P<0.05 for both].As for resistance to paclitaxel, IC50 of MDA-MB-231 breast cancer stem cells with Oct4siRNA interference was significantly decreased [(4.48±0.22) μg/ml] compared with that in the control [(7.99±0.59) μg/ml] and negative siRNA group [(8.10±0.68) μg/ml] (P<0.05 for both).Conclusions MDA-MB-231 breast cancer cells are successfully obtained by serum-free culture. The proliferation potential, invasion ability and drug resistance of breast cancer stem cells were down-regulated by Oct4 gene knock-down.

Objective:To analyze the efficacy of high-throughput hemodialysis (HFHD) therapy on the application of serum ferritin (SF), transferrin saturation (TSAT), ferromodultin (Hepc) and soluble transferrin receptor (sTfR) levels in maintenance hemodialysis (MHD) with renal anemia.Methods:The uremic patients with MHD treatment in the Third People′s Hospital of Hangzhou City from August 2020 to July 2023 were selected as the study object. They were divided into high throughput (30 cases) and low throughput (30 cases) according the treatment methods. The general data; anemia indexes, including red blood cell count (RBC), hemoglobin (Hb), hematocrit (HCT), percentage of reticulocytes (Ret); iron metabolism indexes (SF, TSAT, Hepc and sTfR); inflammations indexes, including β 2-microglobulin (β 2-MG), C-reactive protein (CRP), interleukin-6 (IL-6); renal function indexes, including blood creatinine (SCr), urine creatinine (Cr), blood urea nitrogen (BUN); and adverse reaction were collected. Results:After treatment, the levels of RBC, Hb, HCT, SF and TSAT in the high-throughput group were increased compared with those before treatment: (3.33 ± 0.43) × 10 12/L vs. (2.12 ± 0.24) × 10 12/L, (118.08 ± 11.36) g/L vs. (98.23 ± 8.58) g/L, 0.354 ± 0.030 vs. 0.228 ± 0.037, (486.23 ± 68.22) μg/L vs. (149.34 ± 39.62) μg/L, (36.24 ± 5.82)% vs. (18.72 ± 6.14)%, After treatment, the levels of RBC, Hb, HCT, SF and TSAT in the low-throughput group were increased compared with those before treatment: (2.79 ± 0.32) × 10 12/L vs. (2.19 ± 0.27) × 10 12/L, (111.98 ± 9.07) g/L vs. (97.60 ± 8.33) g/L, 0.330 ± 0.036 vs. 0.214 ± 0.037, (332.73 ± 56.35) μg/L vs. (151.25 ± 39.90) μg/L, (22.23 ± 6.60)% vs. (17.97 ± 5.72)%. In the high-throughput group, after treatment compared with before treatment, Ret, Hepc, sTfR, β 2-MG, CRP, IL-6, SCr, Cr and BUN levels all decreased: (1.02 ± 0.58)% vs.(1.64 ± 0.99)%, (71.56 ± 5.67) μg/L vs. (89.56 ± 7.62) μg/L, (395.07 ± 37.10) μg/L vs. (471.37 ± 41.18) μg/L, (8.38 ± 1.94) μg/L vs. (13.79 ± 2.09) μg/L, (1.07 ± 0.23) mg/L vs. (2.28 ± 0.20) mg/L, (11.40 ± 2.84) ng/L vs. (22.74 ± 6.38) ng/L, (351.54 ± 62.05) μmol/L vs. (530.04 ± 85.06) μmol/L, (6.32 ± 1.49) nmol/L vs. (11.52 ± 2.37) nmol/L, (6.75 ± 1.51) mmol/L vs. (18.37 ± 4.52) mmol/L, compared with before and after treatment in low throughput group, Ret value, Hepc, sTfR, β 2-MG, CRP, IL-6, SCr, Cr and BUN levels decreased: (1.40 ± 0.65)% vs. (1.67 ± 0.78)%, (84.33 ± 7.45) μg/L vs. (88.97 ± 7.79) μg/L, (431.20 ± 37.59) μg/L vs. (459.56 ± 42.22) μg/L, (11.35 ± 1.06) μg/L vs. (14.00 ± 1.90) μg/L, (1.57 ± 0.27) mg/L vs. (2.19 ± 0.23) mg/L, (16..85 ± 3.02) ng/L vs. (23.38 ± 6.17) ng/L, (389.48 ± 67.03) μmol/L vs. (531.02 ± 78.07) μmol/L, (9.80 ± 1.33) nmol/L vs. (11.34 ± 2.18) nmol/L, (7.55 ± 1.21) mmol/L vs. (18.37 ± 4.52) mmol/L, In addition, RBC, Hb, HCT, SF and TSAT levels in high-flux group were higher than those in low-flux group ( P<0.05), while Ret, Hepc, sTfR, β 2-MG, CRP, IL-6, SCr, Cr and BUN levels were lower than those in low-flux group, with statistical significance ( P<0.05). There was no significant difference in the incidence of adverse drug effects between the two groups ( P>0.05). Conclusions:HFHD treatment for patients with renal anemia in MHD has better treatment effect than LFHD, which can effectively correct the anemia state of patients, improve the body iron metabolism response, reduce inflammatory response and improve kidney function, and has certain clinical application value.

Objective: To analyze the clinical effect of low molecular weight heparin calcium(LMWH) combined with aspirin in preventing portal venous thrombosis (PVT) after devascularization. Methods: Retrospective cohort study was carried out. A total of 61 patients with portal hypertension in Zhongshan Hospital affiliated to Fudan University from December 2014 to April 2017 were included in the study, including 43 males and 18 females, aged 24~74 years. According to anticoagulation methods, the patients were divided into treatment group (31 cases of LMWH combined with aspirin) and control group (30 cases of aspirin). Splenectomy and pericardial devascularization or combined devascularization were performed in both groups. Platelet count, prothrombin time, Child-Pugh score, PVT status and surgical complications were observed after surgery. Results: There were no significant differences for platelet count and prothrombin time between the two groups after 7, 14, 30, 60 and 90 days (P>0.05). Child-Pugh scores at 3, 7, 14, 30, 60 and 90 d after operation of the two groups showed no significant differences (P>0.05). Seven days after the operation, PVT in the treatment group and control group was 7 (22.6%) and 7 (23.3%), respectively. Thirty days after the operation, PVT was detected in 8 patients (25.8%) in the treatment group and 15 patients (50.0%) in the control group. Sixty days after the operation, 6 patients (19.4%) had PVT in the treatment group and 13 patients (43.3%) in the control group. PVT was detected in 4 patients (12.9%) in the treatment group and 12 patients (40.0%) in the control group 90 days after operation. The incidence of PVT in the 30, 60 and 90 days after surgery was lower than that in the control group, with significant difference (P<0.05), and the degree of thrombosis was relatively mild. There were 6 cases (20.0%) of complications in the control group and 6 cases (19.4%) in the treatment group, there were no significant differences between the two groups (P>0.05). Conclusion LMWH combined with aspirin is safe and effective in preventing PVT after devascularization.