OBJECTIVES: To reprogram human placental fibroblasts (HPFs) into chemically induced epithelioid-like cells (ciEP-Ls) using a combination of small-molecule compounds. METHODS: HPFs cultured under normoxic conditions were identified using immunofluorescence assay, PCR and chromosomal karyotyping. Under hypoxic conditions (37 ℃, 5% O₂), HPFs were cultured in a medium containing small-molecule compounds C6TSEDRVAJZ (CHIR99021, 616452, TTNPB, SAG, EPZ5676, DZNep, Ruxolitinib, VTP50469, Afuresertib, JNK-IN-8, and EZM0414), and the cell morphology was observed daily. The expression levels of epithelial cell markers in the induced cells were detected by immunofluorescence, Western blotting and PCR. Chromosomal karyotyping of the induced cells was performed and the induction efficiency was calculated. RESULTS: Before induction, HPFs showed positive expressions of fibroblast surface markers CD34 and vimentin and were negative for epithelial surface markers. PCR results showed high expressions of fibroblast-specific genes S100A4 and COL1A1 in HPFs with a normal human diploid karyotype. After one day of induction, the HPFs underwent morphological changes from a multinodular spindle shape to a round or polygonal shape, which was morphologically characteristic of ciEP-Ls. On day 4 of induction, the cells exhibited high expressions of the epithelial cell markers E-cadherin and Lin28A. RT-qPCR results also showed that the cells expressed the epithelial markers Smad3, GLi3, PAX8, WT1, KRT19, and KRT18 with significantly down-regulated expressions of all the fibroblast surface markers and a normal human diploid karyotype. The reprogramming efficiency of HPFs into ciEP-Ls ranged from (64.53±2.8)% to (68.10±3.6)%. CONCLUSIONS The small-molecule compound combination C6TSEDRVAJZ is capable of inducing HPFs into ciEP-Ls under hypoxic conditions with a high induction efficiency.
Humans ; Fibroblasts/drug effects* ; Pregnancy ; Female ; Cell Differentiation/drug effects* ; Pyrimidines/pharmacology* ; Placenta/cytology* ; Cells, Cultured ; Pyridines/pharmacology* ; Pyrazoles/pharmacology* ; Epithelial Cells/cytology*

Objective To determine the expression and clinical significance of pseudogene FMO6P in gastric cancer,and explore its functions and underlying molecular mechanism in regulating the invasion and metastasis of gastric cancer cells.Methods The expression level of FMO6P in gastric cancer tissues and cell lines was detected by quantitative real time polymerase chain reaction(qRT-PCR).The migration and invasion abilities of gastric cancer cells were detected by transwell assay.The effect of FMO6P on the tumor formation and peritoneal dissemination of gastric cancer cells were evaluated by injecting FMO6P-overexpressing gastric cancer cells into the subcutaneous or peritoneal cavity of nude mice respectively.The expression levels of epithelial-mesenchymal transition(EMT)markers,including E-cadherin,N-cadherin,ZEB1,MMP2,and the activation of AKT/mTOR pathway in FMO6P-overexpressing or knockdown gastric cancer cells were measured by Western blot.Results The expression of FMO6P was significantly reduced in tumor tissues compared to its adjacent non-tumor tissues of gastric cancer,FMO6P expression level in tumor tissues was correlated with tumor size and TNM stage.Overexpression of FMO6P significantly inhibited the invasion and migration abilities of gastric cancer cells,while downregulation of FMO6P expression promoted the invasion and migration ability of gastric cancer cells.Overexpression of FMO6P in gastric cancer cells significantly inhibited the subcutaneous tumor formation and peritoneal dissemination of gastric cancer cells in nude mice.Moreover,overexpression of FMO6P promoted the expression of E-cadherin,and inhibited the expression of N-cadherin,ZEB1,and MMP2 in gastric cancer cells.The phosphorylation levels of AKT and mTOR were also downregulated in gastric cancer cells overexpressing FMO6P.Conclusions All these findings suggested that pseudogene FMO6P suppresses the invasion and migration potential of gastric cancer cells in vitro and in vivo,which is possibly through the inhibition of the AKT/mTOR signaling pathway.

Circulating tumor DNA (ctDNA) is cell-free DNA released by tumors or circulating tumor cells, containing abundant tumor-specific information that can serve as biomarkers for cancer early screening, monitoring, prognosis, and prediction of treatment response. This is particularly attractive in the field of gastric cancer, where high-quality screening, monitoring, and prediction methods are currently lacking. Gastric cancer exhibits significant tumor heterogeneity, with large differences in genetic and epigenetic characteristics among different subgroups. Methylated ctDNA has high sensitivity and specificity, which can help clarify tumor genotyping and facilitate the formulation of precise diagnostic and therapeutic strategies. Furthermore, numerous studies have confirmed the unique advantages of methylated DNA in predicting treatment response, adjuvant therapy, and drug resistance assessment, which may be used in the future to enhance the efficacy of chemotherapy regimens and improve patient chemotherapeutic response, and even treat multidrug resistance. However, there are several challenges associated with methylated ctDNA, such as low sensitivity and specificity at single-target sites, limited association between some gastric cancer subtypes and ctDNA, off-target risks, and the lack of large-scale and high-quality clinical research evidence. This review mainly summarizes current research on the methylation status of ctDNA in gastric cancer and connects these findings to early screening, recurrence monitoring, and potential treatment opportunities for gastric cancer. With advances in technology and the deepening of interdisciplinary research, ctDNA detection will reveal more disease information and become an essential foundation for gastric cancer research and precision medicine treatment.

Circulating tumor DNA (ctDNA) is cell-free DNA released by tumors or circulating tumor cells, containing abundant tumor-specific information that can serve as biomarkers for cancer early screening, monitoring, prognosis, and prediction of treatment response. This is particularly attractive in the field of gastric cancer, where high-quality screening, monitoring, and prediction methods are currently lacking. Gastric cancer exhibits significant tumor heterogeneity, with large differences in genetic and epigenetic characteristics among different subgroups. Methylated ctDNA has high sensitivity and specificity, which can help clarify tumor genotyping and facilitate the formulation of precise diagnostic and therapeutic strategies. Furthermore, numerous studies have confirmed the unique advantages of methylated DNA in predicting treatment response, adjuvant therapy, and drug resistance assessment, which may be used in the future to enhance the efficacy of chemotherapy regimens and improve patient chemotherapeutic response, and even treat multidrug resistance. However, there are several challenges associated with methylated ctDNA, such as low sensitivity and specificity at single-target sites, limited association between some gastric cancer subtypes and ctDNA, off-target risks, and the lack of large-scale and high-quality clinical research evidence. This review mainly summarizes current research on the methylation status of ctDNA in gastric cancer and connects these findings to early screening, recurrence monitoring, and potential treatment opportunities for gastric cancer. With advances in technology and the deepening of interdisciplinary research, ctDNA detection will reveal more disease information and become an essential foundation for gastric cancer research and precision medicine treatment.

Objective To determine the feasibility of neck vessel wall imaging technology with three?dimensional variable?flip?angle turbo spin?echo (3D T1w?SPACE) for the detection of carotid atherosclerotic disease before revascularization. Methods Thirty?one patients who underwent carotid endarterectomy (CEA) and fifty?three patients who underwent carotid stenting (CAS) were enrolled prospectively. Neck vessel wall imaging examination were performed in all patients whilecarotid artery DSA were performed in all CAS patients. Quantitative measurements including stenosis, lesion length, and the presence or absence of plaque ulceration obtained with 3D T1w?SPACE and DSA were independently determined. And images of the 3D T1w?SPACE were compared with corresponding histology to identify major plaque components including intraplaque hemorrhage (IPH), lipid rich necrotic core (LRNC), and calcification (CA). The consistency rate, sensitivity, specificity, positive predictive value and negative predictive value were used to assess diagnostic value. Bland?Altman plots, intraclass correlation coefficient (ICC), and Cohen Kappa were determined. Results DSA was served as the reference standard. There was an excellent correlation between 3D T1w?SPACE and DSA images in measuring stenosis (r=0.984, P<0.01) and luminal stenosis [ICC=0.98 (95% confidence interval: 0.96-0.99)]. Bland?Altman plots showed that the two examinations were in good consistency in evaluating the extent of stenosis. Sensitivity (89.5%) and specificity (95.1%) was high in 3D T1w?SPACE images compared to DSA for the detection of ulcers. The consistency rate between 3D T1w?SPACE images and histological results for IPH, LRNC and CA detection were 85.7%, 82.1% and 92.9%, respectively. Sensitivity and specificity were 90.0% and 75.0% for IPH;83.3% and 80.0% for LRNC; 91.3% and 100.0% for CA respectively. However, lesion length measurements by using 3D T1w?SPACE were longer than those measured by using DSA (P<0.01).Conclusion Neck vessel wall imaging technology with 3D T1w?SPACE is a noninvasive and accurate technique for the diagnosis of carotid artery atherosclerotic disease before revascularization.

Objective: To investigate the expression of Flotillin-2 (Flot-2) protein in gastric cancer tissues and its relationship with clinicopathological features and prognosis of gastric cancer (GC) patients. Methods: 112 samples of gastric cancer tissue and the corresponding paracancerous tissue that resected at the gastrointestinal surgery department of the Second Affiliated Hospital of Nanchang University between January 2009 andApril 2010 were collected for this study. The expression of Flot-2 protein in tumor tissues was detected by immunohistochemistry. The survival data were analyzed by Kaplan-Meier and Log-Rank test, and the survival curve was plotted. Spearman correlation analysis was used to examine the relationship between Flot-2 protein expression and clinicopathological characteristics and prognosis of GC patients. Results: In gastric cancer tissues, Flot-2 was primary stained in cytoplasm. Level of Flot-2 was significantly higher in gastric cancer tissues compared with that in paracancerous tissues (53.57% vs 46.43%, P<0.05). Expression of Flot-2 in tumor tissues was significantly associated with tumor size, depth of invasion, lymph node metastasis, distant metastasis and AJCC stage (all P<0.01), but not with gender, age, differentiation degree and tumor location (P>0.05). Moreover, survival analysis showed that the overall survival of patients with low Flot-2 expression was significantly higher than that of the patients with high level (P<0.01). Cox regression analysis indicated that distant metastasis, AJCC stage and Flot-2 expression were the independent risk factors for the prognosis of GC patients. Conclusion: Flot-2 protein was highly expressed in gastric cancer tissues and closely correlated with the poor prognosis of GC patients; Flot-2 is an independent risk factor for GC prognosis and may be served as a potential therapeutic target for gastric cancer.

Objective To explore the clinical causes and preventive measures of complicating ascites of mini-percutaneous nephrolithotripsy (MPCNL).Methods Retrospective analysis of 285 patients with MPCNL for upper urinary tract calculus,which were divided into ascites group and no-ascites group.Results All the procedures were successful.Ascites group of 21 cases,no-ascites group of 264 cases.Univariate analysis showed that the diameter and number of calculus,perfusion pressure,perfusion time,pressure volume of irrigation fluid,preoperative upper urinary tract infection,history of treatment associated with complicating ascites (P＜ 0.05),with age,gender,body mass index no correlation (P＞ 0.05).Logistic regression analysis showed that perfusion pressure,perfusion time,pressure volume of irrigation fluid was independent risk factors after MPCNL concurrent ascites (P ＜ 0.05).Conclusions MPCNL concurrent ascites are closely related to the large perfusion volume,the long operative perfusion time,the high perfusion pressure of irrigation fluid.On the premise of keeping the operative visual field clear,as far as possible to reduce the perfusion pressure,control irrigation fluid-flow rate,reduce the large peffusion volume.These could decrease the coincidence of the ascites.

Objective To study the clinical efficacy of anus-preserving rectectomy by using telescopic anastomosis of colon and rectal mucosa for the middle and lower rectal cancer. Methods A retrospective analysis was carried out in 402 cases with middle and lower rectal cancer undergoing telescopic anastomosis for anus-preserving procedure, including 241 males and 161 females, age ranging from 21 to 99 years, averaging at 55. 7 years. The distal margins of the tumors were within 6 - 9 cm to anal verge. According to TNM staging, there were 123 cases in Stage Ⅰ , 244 cases in Stage Ⅱ , 31 cases in Stage Ⅲ,and 4 cases in Stage Ⅳ. Results 345(345/402, 85. 8% ) cases were followed up, the median time of the follow-up was 6. 1 years. Postoperative complications included 17(4.2%) cases of stomal leakage, 11(2.7% ) cases of stomal stenosis. All patients recovered normal defecating function 12-24 weeks post operation. Local recurrence rate was 6. 3% (22/345). Hepatic and lung metastasis was 13. 6% (47/345) and 2. 6% (9/345)respectively. The five year survival rate was 68. 7% (112/163). Conclusions Anuspreserving rectectomy by using telescopic anastomosis is safe and effective procedure to treat middle and lower rectal cancer, with the preservation of anal function and without the increasing rate of local recurrence.

Objective To evaluate the efficacy of ~(125)I radioactive seeds implantation and ~(125)I plaque brachytherapy for oral carcinoma. Methods Eighteen patients with oral carcinoma confirmed by cytology or histopathology were included in this study, Twelve patients with tongue cancer and six patients with gingival carcinoma, there were 20 ulcerative lesions and 10 metastatic cervical lymph nodes. The mean diameter is (2.3±0.7)cm and(2.8±1.7)cm respectively. The patients were treated with both interstitial implantation of ~(125)I seeds and ~(125)I plaque brachytherapy or with ~(125)I plaque brachytherapy only according to patient's individual conditions.The metastatic cervical lymph nodes were treated with CT-guided interstitial implantation of ~(125)I seeds.The sizes of ulcerative lesions and lymph nodes were observed at 1,3,6 months following treatment,and statistical analysis of the sizes of ulcerative lesions were evaluated by paired t-test.Results After 1,3,6 months follow-up,The mean diameters of ulcerative lesions were(2.1±0.6)cm(t=3.559,P＜0.01),(1.7±0.5)cm(t=7.609,P＜0.01),(0.7±0.6)cm(t=11.508,P＜0.01),the cervical lymph nodes showed reduced size. Furthermore, PET-CT images showed a significant decrease in the metabolic activity of treated tumor. After six months, the focus of infection were healing in 8 patients, the cervical lymph nodes of one patient relapsed after ~(125)I implantation again. Patients were followed for 7 to 28 months, all patients were still alive. Conclusion Interstitial ~(125)I radioactive seeds implantation and ~(125)I plaque brachytherapy provide an effective, safe treatment for oral cancer.

Objective To study clinical therapeutic effects of anus-preserving operation with resecting anal intersphincter to treat ultra-low rectal cancer through abdominal cavity. Methods We retrospectively analyzed 52 cases of ultra-low rectal cancer, with the inferior border of the cancers within 2 cm to anocutaneous line or 5 cm to the edge of anus treated by anus-preserving operation with resecting archos internal sphincter muscles through abdominal cavity and anus. There were 29 males, and 23 females, with age 28 to 76 years old, averaging 56. 3 years old. The inferior border of the cancer were within 4 cm to the edge of anus in 18 cases, including 6 cases of adenoma cancerization, and 5 cm to the anus in 34 cases. Pathologic diagnosis was well-differentiated adenocarcinoma in 21 cases, moderately differentiated in 29 cases, low differentiated in 2 cases, there were 6 cases with adenoma cancerization. 28 cases were Dukes A stage, and 24 B stage. Results The follow-up rate was 88. 4% (46/52), and the median time was 5.9 years. 2 case developed stoma leak (3.8%), and 3 developed stoma stenosis(5.7% ) after operation. The anus could roughly control defecation in 6 ～ 12 mouths after operation. The local recurrence rate was 5.7%, and the 5-year-survival rate was 72.7%. Conclusion By anus-preserving operation with resecting archos internal sphincter muscles, defecation controlling was well reserved by anus, and the 5-year-survival rate was not cut down. This operation is one of the safe and effective operations of anus-preserving procedure.