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Objective: A variety of interstitial cells in tumor microenvironment (TME) based on glioma stem cells(GSC) have the function to promote malignant progression of tumors, but whether these interstitial cells have already undergone malignant transformation and their related molecular characteristics are still poorly understood. Methods: Human SU3-RFP glioma stem cells (GSC) stably transfected with red fluorescent protein (RFP) and interstitial cells from enhanced green fluorescent protein (EGFP) transgenic nude mice were co-cultured in vitro. SU3-RFP cells were also inoculated in different tissues of EGFP-Balb/c nude mice. Immortal EGFP(+) cells were monocloned either from co-culture cells in vitro, or from their xenografts in vivo. These immortal EGFP(+) cells were confirmed to bear characteristics of tumor cell via chromosomal analysis and tumorigenicity assay. Related molecular phenotypes of these cells were further detected through RT-PCR, flow cytometry and immunochemistry(IHC) techniques. Results: (1) Two EGFP(+) cell lines were obtained in vitro, and 5 EGFP(+) cell lines were obtained in vivo tumorigenic experiments. Seven EGFP(+) cell lines all have characteristics of self-renewal, heteroploid of chromosomes and 100% tumorigenicity. (2) Cell surface marker analysis showed cell origin of these cell lines were macrophages (tMΦ1 and tMΦ2 ), dendritic cells (tDC1 and tDC2), fibroblasts (tFB), oligodendrocytes (tOG) and BMSC cells (tBMSC), respectively. (3)All of these seven cell lines co-expressed Sca-1 and c-myc, and have Sox-2 or Nanog expression also, which suggest that they may bear molecular characteristics of mesenchymal stem cells or pluripotent stem cells. Conclusions: (1) Tumor stromal cells in TME have undergone malignant transformation, which is related to the tissue remodeling of TME by GSCs, and not limit to the specific type of their parasitic tissues. (2) Tumor cells originated from GSC and tumor interstitial cells, respectively, are two major types of tumor cells with different origins in glioma parenchyma, can not be simply regarded as tumor heterogeneity, transformed interstitial cells of TME may have the potential to serve as new targets for target diagnosis and therapy.

Human facial morphology is one of the important visible biological characteristics. Understanding the genetic basis underlying facial shape traits has important implications in population genetics, developmental biology, and forensic science. This study extracted 136 Euclidean distance phenotypes from 17 facial features of high-resolution 3D facial images in 1177 Chinese Han adult males. Based on 3× low-depth sequencing data, linear regression was used to analyze the correlation between 125 reported SNPs significantly associated with facial morphology and 136 facial phenotypes. As a result, a total of twelve SNPs from ten genes demonstrated significant association with one or more facial shape traits after adjusting for multiple testing (significance threshold P < 1.35 × 10 -3 ) , together explaining up to 3.89% of age-, and BMI-adjusted facial phenotype variance. These included TEX41 rs17479393, PAX3 rs974448, RAB7A/ACAD9 rs2977562, DCHS2 rs9995821, DCHS2 rs2045323, C5orf50 rs6555969, SUPT3H/RUNX2 rs1852985, MSRA rs11782517, EYA1 rs10504499, GSC rs2224309, DICER1 rs7161418 and DHX35 rs2206437.These results revealed the genetics basis of facial morphology of Han Chinese population, and provided reference data for DNA-based face prediction.

Objective: To observe the lipid metabolism characteristics of tumor-associated macrophages (TAM) after malignant transformation in the glioma micro-environment, and analyze the biological phenotype changes and regulatory mechanisms after inhibiting the lipid metabolism remodeling. Methods: Twelve male Balb/c mice of 6-8 weeks were used in the study. Macrophages (Mφ) were derived from mouse bone marrow, and malignantly transformed macrophages (tMφ 1 and tMφ 2 ) were cloned from the model of glioma stem cell (GSC) through interaction with Mφ in vivo and in vitro . Intracellular lipid droplet formation and cellular cholesterol content were measured respectively in Mφ, tMφ 1 and tMφ 2 . qRT-PCR was performed to detect the genes expression level related with lipid metabolism, including sterol regulatory element binding protein (SREBP), fatty acid synthase (FASN), and 3-hydroxy-3-methylglutarate monoacyl coenzyme A reductase (HMG-CoA). Simvastatin (SIM) was used to analyze the proliferation, immigration and invasiveness ability in tMφ 1 and tMφ 2 after inhibition of the lipid metabolism. Differential expression profiles of miRNAs after SIM treatment were constructed in t-Mφ 1 and bio-informatics analysis was screened and verified for miR449a and its target gene sorting micro-tubule connectin 17 (SNX17) associated with lipid metabolism remodeling. The effect on SNX17 by up-regulated miR-449a were analyzed by qRT-PCR and Western blot, meanwhile, the biological phenotype and cholesterol content were observed after up-regulation of miR449a. Low-density lipoprotein receptor (LDLR) protein levels after SNX17 knockdown and intracellular cholesterol content after LDLR knockdown were detected respectively. Results: The numbers of intracellular lipid droplet formation in tMφ 1 and tMφ 2 were more than that in Mφ ( P <0.001). Likewise, the relative contents of cholesterol (3.89±0.68 and 3.56±0.53), SREBP (4.78±0.60 and 2.84±0.41), FASN (4.65±0.70 and 3.01±0.45), and HMG-CoA (5.74±0.55 and 2.97±0.34) were significantly higher in tMφ 1 and tMφ 2 than those of Mφ (1.01 wel, 1.02 wel and 0.99 wel, respectively) (all P<0.001). The proliferation rates of tMφ 1 and tMφ 2 decreased from (47.06±5.88) % and (45.29±5.64)% to (23.53±4.70)% and (18.74±5.76)%, respectively after treatment with SIM (both P <0.05). The numbers of migrated cells decreased from 1 025±138 and 350±47 to 205±63 and 99±25, respectively (both P <0.001). And the numbers of invasiveness cells decreased from 919±45 and 527±34 to 220±23 and 114±21, respectively (both P <0.001). While the relative intracellular cholesterol content decreased to 0.52±0.08 and 0.58±0.07 (both P <0.05), respectively. MiR-449a was screened from tMφ 1 by SIM, and the target gene was analyzed and verified to be SNX17. SNX17 expression was down-regulated, and the proliferation rate, the number of migration and invasiveness was significantly decreased after miR-449a over-expression (all P <0.05). Low-density lipoprotein receptor (LDLR) expression was down-regulated after knock-down of SNX17, while the cholesterol content was decreased after knock-down of LDLR in tMφ 1 and tMφ 2 (all P <0.05). Conclusions: Malignantly transformed TAMs undergo lipid metabolism remodeling characterized with enhanced lipid metabolism. MiR-449a regulates the LDLR by targeting SNX17, thereby affecting the lipid metabolism of malignantly transformed macrophages, and subsequently inhibiting its proliferation, migration, and invasion ability. Precise intervention with miR-449a/SNX17/LDLR axis could provide an experimental basis for reversing its tumor-promoting micro-environment remodeled by GSC through metabolic intervention.

This study aims to comprehensively summarize articles on the treatment of coronary heart disease angina pectoris with heart blood stasis syndrome(CHD-AP-HBSS) by Guanxin Shutong Capsules(GSC), and comprehensively evaluate the evidence and value of the formula in &quot;6+1&quot; dimensions based on multi-criteria decision analysis(MCDA) model and Guidelines for Management of Comprehensive Clinical Evaluation of Drugs(trial version 2021) with quantitative and qualitative methods, which is expected to highlight the clinical characteristics and advantages of the prescription and serve as a reference for medical and health departments. The dimensions are grade A, B, C, or D. In terms of safety, according to the reports from the spontaneous reporting system of National Center for ADR Monitoring, and literature analysis, GSC mainly results in the adverse reactions of mild or moderate nausea, diarrhea, rash, palpitation, and headache, with favorable prognosis in patients. Nevertheless, severe adverse reactions have been reported after marketing. Thus, additional evidence for its safety should be accumulated, and the current safety is grade B. Meta-analysis proves that GSC in combination with conventional treatment is superior to conventional treatment alone in alleviating AP and symptoms indicated by electrocardiogram(ECG). Therefore, the effectiveness of the medicine is grade A. As for the pharmacoeconomic value, based on the assumption that the per capita disposable income in 2020 is the willingness-to-pay threshold, it is more economical for CHD-AP patients to use GSC combined with conventional treatment than conventional treatment alone. However, as the currently available clinical parameters fail to support the difference in indirect cost between the two groups, the accuracy in the cost of intervention plan needs to be further improved. Therefore, the economy of the prescription is grade B. GSC has been awarded 13 national invention patents and 1 utility model patent, and won a lot of national and provincial patent awards, marking the enterprise service awareness and innovativeness. As a result, it is grade A in aspect of innovation. A questionnaire on the suitability of GSC suggests that publicity and promotion influence patients' choice and thus additional efforts should be made in this aspect. The suitability of this formula is grade B. Both rural and urban residents can afford the medicine for the whole course and the price is close to that of similar Chinese patent medicines. In addition, it is accessible regardless of season and place, so accessibility is grade A. As a Mongolian empirical formula destined to treat heart stabbing pain, it has the effects of activating blood and resolving stasis, dredging channels and activating collaterals, and moving Qi to relieve pain, and boasts the empirical evidence of more than 2 000 human cases. With prominent characteristics of traditional Chinese medicine, it is grade B. Based on the MCDA model and CSC v2.0, GSC is comprehensively class A in the treatment of CHD-AP-HBSS. The result can serve as a reference for basic clinical medication management.

Objective: To investigate and compare the clinicopathological characteristics of gastric stump cancer(GSC) and the recurrent of gastric cancer (RGC) following radical gastrectomy, and to evaluate survival prognosis., Methods: A retrospective cohort study was performed on clinicopathological and survival data of patients with GSC (n=31) and with RGC (n=105) following radical gastrectomy at the Chinese People's Liberation Army General Hospital between January 1992 and August 2017. GSC was defined as cancer occured in remnant stomach ≥10 years after radical gastrectomy, while RGC was defined as <10 years. Patients of both groups received radical resection or palliative operation with tumor resection and had complete clinicopathological data regarding the first operation and gastric stump operation. T-test was used to compare quantitative data between the two groups, and Pearson χ 2 test was used to compare qualitative data between the two groups. Kaplan-Meier method was applied to draw survival curves and log-rank test to assess survival differences., Results: Of the 136 enrolled patients, 113 were male and 23 were female. In the first operation, compared with RGC group, in GSC group, the Borrmann type and histological differentiation were more better [Borrmann I(: 11/31 (35.5%) vs. 5/105 (4.8%), χ 2 =23.003, P=0.001; the high differentiation: 15/31 (48.4%) vs. 1/105 (1.0%), χ 2 =57.137, P=0.001]; the tumor diameter was smaller [<4 cm: 28/31(90.3%) vs. 56/105(53.3%), χ 2 =14.045, P=0.001]; the pT stage [pT1: 12/31 (38.7%) vs. 3/105 (2.9%), χ 2 =50.373, P=0.001], pN stage [pN0: 28/31 (90.3%) vs. 19/105 (18.1%), χ 2 =55.722, P=0.001] and pTNM staging [I(: 26/31 (83.9%) vs. 11/105 (10.5%), χ 2 =66.688, P=0.001] were earlier. Most of the GSC occurred at non-anastomotic sites, while the recurrence mostly occurred at anastomotic sites [51.6%(16/31) vs. 61.9%(65/105), χ 2 =7.520, P=0.023]. Compared with RGC group, GSC group had better histological differentiation [high differentiation: 5/31 (16.1%) vs. 2/105(1.9%), χ 2 =10.029, P=0.007]. There was more histological type change between the first and the second operation in GSC group than that in RGC group[48.4%(15/31) vs. 26.7%(28/105), χ 2 =5.222, P=0.022]. The overall survival time of GSC group was significantly longer than that of RGC group [mean: (161.0±18.6) months vs. (50.8±27.6) months, respectively, Log-rank: 76.818, P=0.001]. The survival time after the second surgery of GSC group was longer than that of RGC group [mean: (30.7±18.4) months vs. (20.5±15.0) months, P=0.003]. In the subgroup analysis of all the 136 patients according to histological type change between the two surgeries (unchanged 93 patients, changed 43 patients), compared with unchanged group, the overall survival time of changed group was longer [mean: (99.6±56.5) months vs. (72.1±58.1) months, P=0.008]., Conclusions: GSC patients have better histological differentiation and earlier clinical stage of primary gastric cancer, and longer survival time compared with RGC patients. The histological type change between two operations may be used as a new factor to define GSC.

Due to different disease background of gastric stump cancer(GSC) patients (benign or malignant lesion, reconstruction of digestive tract, etc.), the GSC surgical procedure and the difficulty of lymphadenectomy are also different. The extent of radical lymphadenectomy for gastric stump cancer should extend beyond D2 lymphadenectomy, according to the different backgrounds of initial disease, reconstructions, and tumor location. A lymphadenectomy should include the lymph nodes in anterior (No.17) and posterior (No.13) surface of the pancreatic head, along the superior mesenteric vein (No.14v), jejunal mesenteric lymph nodes, the splenic hilum (No.10), infra-diaphragm (No.19), esophageal hiatus of the diaphragm(No.20), para-esophageal nodes in the lower thorax (No.110) and supra-diaphragm (No.111). To achieve R0 resection, combined adjacent organ (transverse colon and spleen) resection is necessary in some cases. The indication for endoscopic submucosal dissection of early remnant gastric cancer is similar to primary early gastric cancer in selected patients. The advantages of laparoscopic and robot-assisted surgery for early remnant gastric cancer need to be confirmed through multicenter studies. Compared with primary proximal gastric cancer, remnant gastric cancer showed an equivalent survival rate. Roux-en-Y is the first choice for digestive tract reconstruction.

Objective: To study the clinicopathological features and prognosis of gastric stump cancer (GSC) following subtotal gastrectomy for gastric cancer, to compare the clinicopathologic differences between narrow GSC and generalized GSC, and to compare the prognosis between GSC and primary proximal gastric cancer (PPGC) after radical resection., Methods: Literatures of GSC-associated clinical study were searched by computer from the Cochrane Library, Medline, PubMed, CNKI, Wanfang and VIP databases, and the retrieval period was from the establishment of database to December 31, 2017., Inclusion Criteria: (1) GSC was defined as a carcinoma arising in the gastric remnant after radical gastrectomy for gastric cancer, and confirmed by the pathological or histological examination, the elapsed time from the initial operation was not considered in the definition. (2) Retrospective or prospective clinical cohort study. (3) Study included at least one of below items: gender, anastomotic type in gastric cancer surgery, the interval between the initial surgery and diagnosis of GSC, the location, treatment, pathological differentiation, pathologic stage, lymph node metastasis rate and prognosis of GSC. (4) When similar studies were reported by the same institution or author, either the better quality study or the newest publication was chosen., Exclusion Criteria: (1) Abstracts, reviews, case reports, meeting record, editorials and repeated research. (2) Studies including patients with initial non-gastric cancer. In this study, gastric stump cancer(GSC) after gastric cancer was divided into two groups: the incidence without limit interval time (generalized GSC group) and above 10 years (narrow GSC group). Selective trials were Meta-analyzed by the Stata13.0 software and statistical analysis was performed using SPSS 21.0 software., Results: A total of 27 literatures were finally enrolled, which comprised 1463 GSC patients, including 1146 males and 317 females. The generalized group and narrow GSC group had 921 and 542 patients respectively. The generalized GSC group and the narrow GSC group did not significantly differ in terms of previous reconstruction mode, types of differentiation, pathologic T staging, postoperative pathology tumor-node-metastases staging, and distant metastasis rate (χ 2 =2.341, 0.926, 0.350, 0.965, 2.311 respectively, all P>0.05). As compared to generalized GSC group, narrow GSC group had higher ratio of male patients (82.8% vs. 75.7%, χ 2 =9.909, P=0.002), more lesions locating in anastomotic stoma (37.8% vs. 26.1%, χ 2 =18.091, P=0.000), higher ratio of patients undergoing radical resection (84.2% vs. 70.3%, χ 2 =11.738, P=0.001), higher positive rate of postoperative lymph node (45.8% vs. 34.5%, χ 2 =6.319, P=0.012), and larger size of tumor [(5.9±2.2) cm vs. (4.5±1.9) cm, t=9.151, P=0.000]. The overall 5-year survival rate and postoperative pathology stage III(-IIII( survival ratio in narrow GSC group were higher compared to general GSC group (42.7% vs. 30.6% and 27.5% vs. 18.1%, respectively), which were significantly different (χ 2 =10.938, P=0.000; χ 2 =4.128, P=0.042), while the postoperative pathology stage I(-II( survival ratio was not significantly different between two groups (67.3% vs. 67.0% respectively, χ 2 =0.015, P=0.92). There was no significant difference in the 5-year survival rate between GSC with radical resection and PPGC(RR=1.04, 95%CI:0.79-1.36, P=0.805) and the 5-year survival rate of same postoperative pathology stage was not significantly different between two groups (I(-II( stage: RR=1.08, 95%CI:0.93-1.26, P=0.328; III(-IIII( stage: RR=0.59, 95%CI:0.33-1.04, P=0.111)., Conclusions: There are some different clinicopathological features between the generalized and the narrow GSC after gastric cancer surgery. The prognosis of GSC after radical resection is similar to primary proximal gastric cancer.

Gastric stump cancer (GSC) is a carcinoma arising from the remnant stomach following gastric surgery for benign or malignant disease, and is more common in men. The risk of morbidity has an obvious time dependence. GSC incidence is likely to rise with lengthening of the initial operation interval. The GSC time interval after malignant disease is significantly shorter than that of benign disease. GSC etiologies mainly include duodenogastric reflux and denervation of the gastric mucosa resulting in the change of the gastric environment after gastrectomy and the Helicobacter pylori infection. Due to atypical clinical symptoms, GSC is always identified at an advanced stage and the long-term survival rate is low. An optimal endoscopic surveillance system is essential to improve early detection rates. Treatments in GSC and primary gastric cancer are the same and include resection of the lesion and radical lymph node dissection. R0 resection is an important prognostic factor. Here we review previous reports with respect to epidemiological characteristics, etiology, clinical symptoms, treatment, and prognosis of GSC.

Gastric stump cancer(GSC) is defined as newly developed remnant stomach cancer following gastrectomy. This definition initially referred to carcinoma detected in the remnant stomach more than 5 years after the primary surgery for a benign disease. Subsequently, this timeframe was extended to 10 years after the primary surgery for a malignant disease. Recently, the concept of "carcinoma in the remnant stomach(CRS)" proposed by the Japanese Gastric Cancer Association was introduced in China. The new definition encompasses all carcinomas arising in the remnant stomach following gastrectomy, irrespective of the histology of the primary lesion, extent of resection, or reconstruction method. It includes all carcinoma types that have developed in the remnant stomach, such as newly developed cancer, recurrent cancer, remaining cancer, and multiple cancers. Considering the current diagnosis and treatment status of gastric cancer in China, if CRS is to be used as a direct equivalent to GSC in clinical practice, confusion may arise concerning disease identification and diagnosis. Following several discussion rounds, a meta-analysis of the literatures at home and abroad, and a multicenter national retrospective study with a large sample population, the "Chinese surgeons' consensus opinion for the definition of gastric stump cancer (version 2018)" was completed. By reviewing the detailed evidence-based medicine supporting the consensus document, this paper aims to assist clinical diagnosis and enhance future academic exchange.

Objective: To investigate the tumor microenvironment of immune tolerance induced by glioma stem cells (GSC). Methods: Human GSC SU3 cells transfected with red fluorescent protein (SU3-RFP) gene were implanted into the brain, subcutis (armpit and foot), liver and abdominal cavity of transgenic green fluorescence protein (GFP) nude mice to establish RFP(+) /GFP(+) dual fluorescence solid tumor model. The re-cultured cells derived from implanted tumor tissues, SU3-RFP cells co-cultured with peritoneal fluid of transgenic GFP nude mice and malignant ascites of tumor-bearing mice were observed by fluorescence microscopy and real-time video image tracing to analyze the microenvironment of immune tolerance mediated by RFP(+) /GFP(+) implanted tumor. Results: Dual fluorescence labeled frozen section showed that all of cells in the tumor microenvironment were GFP(+) , while the pressed tissue-patch showed that the tumor blood vessels exhibited a RFP(+) /GFP(+) double-positioning yellow. In the GFP single fluorescence labeled tumor tissue, all of cells in the microenvironment were green, including tumor edge, necrotic foci and blood vessel. Among them, CD68(+) , F4/80(+) , CD11c(+) , CD11b(+) and CD80(+) cells were observed. In the dual fluorescence labeled co-cultured cells, the phagocytosis and fusion between green host cells and red tumor cells were also observed, and these fusion cells might transfer to the malignant dendritic cells and macrophages. Conclusions: The tumor microenvironment of immune tolerance induced by GSC is not affected by the tissue types of tumor-inoculated sites, and the immune tolerance mediated by inflammatory cells is associated with the inducible malignant transformation, which may be driven by cell fusion.

Objectives: To investigate the clinical feature and surgical procedures of gastric stump carcinoma (GSC) and to identify the prognostic factors which influence survival rate of GSC patients., Methods: Clinical data of 167 patients who underwent R0 resection for gastric stump carcinoma at Chinese People's Liberation Army General Hospital between January 1990 and December 2012 was collected. There were 144 male and 23 female cases. The clinicopathological features of GSC patients were compared between those who underwent initial surgery for benign disease (GSC-B group, 78 cases) and for gastric cancer (GSC-M group, 89 cases). The analysis of therapeutic methods and survival time were also performed.t-test was used to compare the quantitative data between two groups. Pearson χ(2) test was used to compare the various clinicopathological characteristics between the two groups. Kaplan-Meier method was used to analyze the survival rate. Multivariate survival analysis was based on the Cox proportional hazard model., Results: Compared with GSC-M group, the interval time between initial gastrectomy and surgery in GSC-B group was longer ( (28.2±10.2) years vs. (10.8±1.0) years, t=15.902, P=0.001). There were 56 patients (71.8%) who received BillrothⅠ reconstruction in GSC-B group, and 49 patients (55.1%) who received BillrothⅡ reconstruction in GSC-M group, the difference of anastomosis method between the two groups was statistically significant (χ(2)=25.770, P=0.001). Compared with GSC-M group, the tumor of GSC-B group was usually located at the anastomotic site (χ(2)=6.975, P=0.031). The overall 1-, 3-, and 5-year survival rates of the 167 patients were 87%, 60%, and 41%. The 5-year survival rates for TNM stagesⅠ, Ⅱ, and Ⅲ were 65%, 43%, and 22%, respectively (P= 0.001). Multivariate analysis showed that small intestinal or esophageal infiltration (HR=1.957, 95%CI: 1.096 to 3.494, P=0.023), tumor location (HR=1.618, 95%CI: 1.104 to 2.372, P=0.014), and TNM stage (HR=2.307, 95%CI: 1.708 to 3.118, P=0.001) have independent effect on survival. The metastasis rates of perigastric lymph nodes, jejunum anastomosis and mesenteric lymph nodes were very high (56.3% and 65.2%, respectively)., Conclusions: The GSC appears earlier in patients with gastrectomy for malignant disease than those with benign disease. Appropriate curative resection including residual lymph node dissection is very important to improve the prognosis. Small intestinal or esophageal infiltration, tumor location, and TNM stage have independent effect on survival.

Objective: To study the phenotype and tumorigenicity of SHG-44 glioma stem cell spheres and the pathological characteristics of their xenograft tumors., Methods: SHG-44 glioma cells were cultured under neural stem cell medium and glioma stem cell spheres were collected. Immunocytochemistry was used to dectet the expression of CD133, nestin, A2B5, vimentin, VEGFR-2 and IDH R132H. Cell spheres were induced using serum-containing medium, and the expression of CD133, nestin, vimentin, GFAP, β-III tubulin and GalC in the cell spheres were detected. The expression of CD133, nestin, VEGFR-2, GFAP, S-100 and CD34 in the intracranial xenograft tumor tissues was detected using immunohistochemistry. The pathological characteristics of orthotopic xenograft tumors generated from the SHG-44 glioma cells and SHG-44 glioma stem cell spheres were compared., Results: SHG-44 glioma stem cell spheres were collected successfully after cultured under neural stem cell medium. The ratio of CD133(+) cells in the passage 10 SHG-44 glioma stem cell spheres was (71.63 ± 5.92)%, significantly higher than that in the SHG-44 glioma cells [(1.95 ± 1.45)%]. Immunocytochemistry showed that in the SHG-44 glioma cell spheres, the ratio of nestin(+) cells was (84.06 ± 7.58)%, vimentin(+) cells (29.11 ± 3.44)%, VEGFR 2(+) cells (64.44 ± 3.69)%, and A2B5(+) cells (14.08 ± 2.19)%. A subpopulation of cells with mutation of IDH R132H was detected harboring in the SHG-44 glioma cell spheres. After induction of differentiation with serum-containing medium, the ratio of CD133(+) cells was (1.89 ± 1.27)%, nestin(+) cells (6.67 ± 2.75)%, vimentin(+) cells (93.75 ± 2.95)%, GFAP (+) cells (91.33 ± 4.75)%, β-III tubulin(+) cells (82.36 ± 4.02)%, and GalC(+) cells (8.92 ± 3.19)%. Immunohistochemistry showed positive expression of GFAP, S-100, VEGFR-2, and negative of CD133 and nestin in the orthotopic xenograft tumors. A very small amount of human-specific CD34 cells formed a tubular structure. Compared with the SHG-44 glioma cell-formed xenograft tumor, the SHG-44 glioma stem cell-formed xenograft tumor exhibited a higher local invasiveness., Conclusions: SHG-44 glioma cell spheres are successfully collected after cultured under neural stem cell medium. They belong to the CD133(+)A2B5(-) GSC subpopulation, highly expressing VEGFR-2, possess the ability of both self-renewal and multi-directional differentiation, and may participate in the formation of vasculogenic mimicry.

Objective: To explore the effects of B-cell specific Maloney leukemia virus integration site 1 (Bmi1) gene on endothelial cells promoting glioma stem cell (GSC)-like phenotype., Methods: Glioblastoma cell line GL261 and brain micro-vessel endothelial cell line b.END3 were used. Transwell co-culture system, limit dilution assay, xenograft, real-time polymerase chain reaction (PCR), Western blot, fluorescence activating cell sorter (FACS) and gene knock-down assay were used to determine the GSC-like phenotype and Bmi1 gene expression in glioma cells., Results: Compared with the control of GL261 cell alone, (1) more and larger tumor spheres formed after co-culturing with endothelial cells (62.5% ± 1.5% vs 25.0% ± 4.6% at 40 cells/well, P = 0.000). Xenografts generated by GL261 cells with b.END3 cells appeared earlier and were larger than that by GL261 cells alone ((0.798 ± 0.297) cm(3) vs (0.362 ± 0.123) cm(3), P = 0.000); (2) CD133 positive glioma cells increased after co-culturing with endothelial cells (8.48% ± 0.78% vs 4.81% ± 0.37%, P = 0.000); (3) the expression of Bmi1 in co-cultured glioma cells was up-regulated at mRNA level (2.72 ± 0.18 vs 1.00 ± 0.15, P = 0.000) and at protein level; (4) the above phenomenon was attenuated when Bmi1 gene expression was inhibited by siRNA in glioma cells, CD133 positive portion of Bmi1-knockdown GL261 cells co-culturing with b.END3 cells decreased than that of wildtype GL261 cells (0.34% ± 0.21% vs 1.70% ± 0.69%, P = 0.025)., Conclusion: Endothelial cells promote GSC-like phenotype by up-regulating the expression of Bmi1 in glioma cells.

Objective: To explore the effects of all-trans retinoic acid (ATRA) on glioma stem cell phenotype., Methods: The glioma stem cell (GSC) from surgically resected human glioma specimens were isolated and enriched by neurosphere assay and then its differentiation was induced with all-trans retinoic acid (ATRA, 1 µmol/L) for 1 week. Markers were determined by flow cytometry, Western blot and reverse transcription-polymerase chain reaction (RT-PCR). Side population cells were analyzed by flow cytometry. Growth characteristics were detected by neurosphere formation assay and cell cycle analysis. GSC and the differentiated cells (1×10(5)) were implanted stereotactically and intracranially into the Balb/c nude mice to compare the survival time. All data were analyzed with the SPSS software version 17.0., Results: ATRA potently induced the differentiation of GSC and reduced glioma stem cell phenotype. And there were an elevated expression of glial fibrillary acidic protein (GFAP) and a reduced expression of such stem cell makers as CD133 and Nestin. The side population rate decreased. ATRA inhibited the neurosphere formation of GSC and induced the arrest of cell growth. ATRA could prolong the survival time., Conclusion: GSC may be differentiated efficiently by ATRA. The phenotype of GSC decreases obviously after the differentiation of ATRA and the survival time is prolonged. Thus ATRA may be applied for targeted therapies of glioma stem cell.

Objective: To study the effects of Ginseng and Sanchi Compositions (GSC) on the protein and mRNA expressions of Ras, extracellular signal-regulated Kinase1/2 (ERK1/2), and C-Raf-1 of ischemic myocardium of rats with acute myocardial infarction (AMI)., Methods: By adopting myocardial ischemia Wistar rat model with the ligation of the left anterior descending coronary artery, the normal control group, the sham-operation group, the model group, the Betaloc group, the high and low dose GSC group were set up. The protein expressions of Ras, C-Raf-1, ERK1/2,and phosphor-ERK1/2, p-ERK1/2 (p-ERK1/2) were detected by Western blot. The mRNA expressions of Ras, C-Raf-1, ERK1, and ERK2 were detected using Real-time PCR., Results: The protein expressions of Ras, c-Raf-1, and p-ERK1/2 and their mRNA expressions in the model group increased more than those in the normal control group and the sham-operation group (P < 0.05). The protein expressions of Ras, c-Raf-1, and p-ERK1/2 and their mRNA expressions in the high and low dose GSC groups and the Betaloc group were significantly higher than those in the model group (P < 0.05). The protein expressions of Ras, c-Raf-1, and p-ERK1/2 and their mRNA expressions increased more obviously in the high dose GSC group than in the low dose GSC group (P < 0.05). The ERK1/2 protein expression was not significantly different among all groups (P > 0.05)., Conclusions: GSC could up-regulate the protein expressions and mRNA expressions of Ras, C-Raf-1, and p-ERK1/2. It suggested that GSC might promote the angiogenesis through Ras signal transduction pathway dose-dependently.

Objective: To explore the clinicopathological characteristics and prognostic factors of gastric stump cancer(GSC)., Methods: The clinical data of 138 patients with GSC treated from January 1992 to July 2008 were reviewed and analyzed. The patients included 122 males and 16 females with a mean age of 61.5 years, and the mean interval between the initial operation and second diagnosis was 21.9 years., Results: The endoscopy and pathological examination showed Borrmann III/IV in 127 (92.7%) patients and undifferentiated carcinoma in 115 (83.3%) patients. The resectability and radical resectability rate were 72.4% and 59.4%. The 1-, 3- and 5-year survival rates was 59.2%, 30.1% and 14.2%, respectively. The median overall survival time was 19.4 months. Univariate Log-rank test indicated that Borrmann type, histological type, tumor diameter, TNM stage, depth of invasion, number of metastatic lymph node, distant metastasis and option of treatment were significant prognostic factors for GSC. While TNM stage, depth of invasion, distant metastasis and option of treatment were prognostic factors on multivariable analysis. The median survival time of patients underwent radical resection was significantly longer (36 months) than that of patients received palliative resection (8 months, P < 0.05) and chemotherapy only (5 months, P < 0.05). Among patients with a tumor of T4 stage, the median survival time was statistically prolonged by combined evisceration (18.6 months) when compared with the patients received palliative surgery., Conclusions: TNM stage, depth of invasion, distant metastasis and option of treatment are independent prognostic factors for GSC. Early diagnosis and radical resection may play an important role in improving the prognosis of GSC.

Objective: To study the role for piwil2 gene (zili) in the development of the ectoderm, mesoderm and endoderm during early embryogenesis of zebrafish., Methods: zili morpholino antisense oligonucleotide and 5 mis-pair control morpholino were used in this study. zili was cloned into expression vector. zili mRNA was synthesized in vitro. The antisense RNA probes of gsc, evel and sox17 were synthesized. zili-MO, zili-cMO and zili mRNA was microinjected into one-cell embryos, respectively. Whole-mount in situ hybridization was used to monitor the expressions of marker genes., Results: Microinjection of zili-MO, which knocked down the expression of zili, downregulated the expression of the ectodermal and mesodermal marker gene gsc, promoting the expression of the ectodermal marker gene evel and resulting in the decrease of endodermal cell expressed sox17. The overexpression of zili, promoting the expression of gsc, inhibiting the expression of eve1 and resulting in the decrease of endodermal cell expressed sox17 were observed after microinjection of zili-mRNA., Conclusion: zili might have some effect on the formation of the ectoderm, mesoderm and endoderm during early embryogenesis and might be important for normal embryonic development.

Background and Objective: Gastric stump cancer (GSC) has unique biological behaviors and poor prognosis. The surgical treatment for GSC is complex. This study was to explore the clinical characteristics of GSC and the effect of operation patterns on long-term survival, and investigate its prognostic factors., Methods: The clinical data of 81 GSC patients, treated in Zhejiang Cancer Hospital from January 1990 to December 2007, were analyzed. Patients' survival was analyzed by Kaplan-Meier method; the prognosis was analyzed by Cox multivariate regression model., Results: Of the 81 patients, 47 (58.0%) received radical resection, 34 received non-radical operation (including palliative operation, short circuit or fistulation and exploration). The overall 1-, 3-, and 5-year survival rates were 69.1%, 24.3%, and 11.8% in the whole group. The 1-, 3-, and 5-year survival rates were significantly higher in radical resection group than in non-radical operation group (93.6% vs. 35.3%, 42.0% vs. 5.9%, 20.8% vs. 0, P<0.05). Cox multivariate regression analysis showed that clinical stage, pathologic classification, radical resection, peritoneal seeding and liver metastasis were related with the survival of GSC patients., Conclusions: Radical resection may help to improve the prognosis of GSC. Clinical stage, pathologic classification, radical resection, peritoneal seeding and liver metastasis are independent prognosis factors of GSC.

Objective: To assess the efficacy and safety of Ganshao Capsule (GSC) in treating clomiphene (CC)-resistant hyperandrogenemic polycystic ovarian syndrome (PCOS), and to explore the feasibility of using CC for improving ovulation induction after withdrawal of GSC., Methods: Twenty-seven PCOS patients were given GSC for 8 weeks (2 cycles). Changes in serum reproductive hormone, body mass index (BMI), adverse reaction, as well as pelvic ultrasonographic feature were observed before treatment, at the end of 4 weeks and 8 weeks after treatment. After stopping GSC medication, CC was used to induce ovulation in patients whose serum testosterone (T) < (2.1+/-0.8) nmol/L but without ovulation. Condition of follicle growth and effect of GSC were monitored., Results: After 4 weeks of treatment, serum reproductive hormones were significantly changed. At the end of 8 weeks, bilateral ovarian volume, number of follicles, and diameter of follicles were significantly reduced, and endometrium obviously thinned. Within 2 months after stopping medication, 6 in the 27 patients got natural ovulation, and 2 natural pregnancy. In the other 19 patients who received ovulation induction during the 37th cycles, 17 had ovulation in the 25th cycles, 7 got pregnancy, the ovulation rate being 89.5%, ovulation cycle rate 67.6%, pregnant rate 36.8%, and pregnant cycle rate 28.0%., Conclusion: Endometrium, ovarian morphology and BMI got significant improvement in PCOS patients with CC- resistance and hyperandrogenemia. The sensitivity to ovulation induction with CC were also improved.