Objective:To analyze the invasion characteristics and prognostic factors of patients with Masaoka-Koga stage Ⅲ thymoma.Methods:The tumor invasion characteristics of 179 patients who were diagnosed with Masaoka-Koga stage Ⅲ thymoma and treated in Affiliated Cancer Hospital of Zhengzhou University from January 2000 to June 2018 were analyzed retrospectively. According to the treatment methods, all patients were divided into the radical operation group ( n=94), palliative operation group ( n=39) and simple biopsy group ( n=46). The χ2 test was used to compare the classified variables, Kaplan- Meier method was utilized to calculate the cumulative survival rate, log-rank method was used for group comparison and univariate analysis, and Cox’s regression model was used for multivariate analysis. Results:Mediastinal pleural invasion (86.0%) was the most common site, followed by pericardium (50.8%), great vessel (40.8%) and lung (36.3%). The proportion of macrovascular invasion in the radical operation group was 14.9%, significantly lower than 79.5% and 60.9% in the palliative surgery group and biopsy group (both P<0.001). Multivariate analysis showed that the nature of operation ( P<0.001), age ( P=0.011), radiotherapy ( P=0.020) were the independent factors affecting overall survival (OS), while nature of operation ( P<0.001), age ( P=0.004), radiotherapy ( P=0.020), number of invasive organs ( P=0.023) and pathological type ( P=0.016) were the independent factors affecting progress-free survival (PFS). Conclusions:For patients with Masaoka-Koga stage Ⅲ thymoma, mediastinal pleura is the most common site of invasion, pericardium, lung and great vessels are also commonly invaded. The invasion of mediastinal pleura, pericardium and lung exerts slight effect on surgical resectability, whereas great vessel involvement can significantly affect surgical resectability. OS and PFS in patients undergoing radical resection are significantly better than those in patients treated with palliative resection and biopsy. Radical resection is the most important factor affecting prognosis.

OBJECTIVETo investigate whether direct administration of adenoviral vectors (Ad) containing the complementary deoxyribonucleic acid (cDNA) of vascular endothelial growth factor 165 (Ad-VEGF165) induces porcine coronary collateral vessel formation, improves regional myocardial perfusion and function and is safe.

METHODSThree weeks after miniature swine underwent left thoracotomy and placement of an Ameroid constrictor on the left circumflex coronary artery (LCX), Ad-VEGF165 (n = 6) or the control, Ad expressing beta-galactosidase cDNA (Ad-Gal, n = 6), was directly administered into the ischemic myocardium in the circumflex distribution. All animals were sacrificed 4 wk after the second surgery. Myocardial perfusion and function were assessed by electrocardiogram-gated single photon emission computed tomography (GSPECT) imaging. Ex vivo coronary angiography was performed to examine collateral vessels. Toxicity was assessed by blood analyses on the day just before (day 0) and on day 1, 3, 7, 28 after vector delivery and by vascular, myocardial and liver histology after sacrifice.

RESULTSGSPECT imaging 4 wk after administration of Ad-VEGF165 demonstrated significant reduction in ischemic area (P < 0.01) and rest ischemic severity (P < 0.01) and significant improvement in the left ventricular ejection fraction (P < 0.01) and regional wall motion (P < 0.05) compared with that of Ad-Gal and before administration of Ad-VEGF165. Collateral vessel development assessed by coronary angiography was significantly greater in the Ad-VEGF165 group than in the Ad-Gal group (P < 0.05). General safety parameters, including routine blood parameters, liver and kidney function and cardiac specific parameters demonstrated no difference between Ad-VEGF165 and Ad-Gal animals except for the red blood cell count on day 28 (P < 0.05) and blood urea nitrogen on day 7 (P < 0.05). Only transient elevations in creatine phosphokinase (P < 0.05) and aspartate transaminase (P < 0.05) on day 1 were revealed compared with that before vector administration in both groups. Histologically, no atherosclerotic lesion in the circumflex and no inflammation in liver were revealed and only a small myocardial necrosis was observed in one Ad-VEGF165 animal (area < or = 20%) and one Ad-Gal animal (area < 10%).

CONCLUSIONSAd-VEGF165 can induce coronary collateral vessel formation, improve regional myocardial perfusion and function and is safe by means of direct injection, which suggesting that this strategy may be useful in treating human ischemic heart disease.

Adenoviridae ; genetics ; Animals ; Collateral Circulation ; Coronary Angiography ; Coronary Vessels ; physiopathology ; DNA, Complementary ; administration & dosage ; genetics ; Electrocardiography ; Endothelial Growth Factors ; genetics ; physiology ; Female ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Genetic Vectors ; administration & dosage ; genetics ; Lymphokines ; genetics ; physiology ; Male ; Myocardial Ischemia ; diagnostic imaging ; genetics ; therapy ; Neovascularization, Physiologic ; physiology ; Swine ; Swine, Miniature ; Tomography, Emission-Computed, Single-Photon ; methods ; Treatment Outcome ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

Objective: To compare the effect of neoadjuvant chemoradiotherapy (NCRT) and neoadjuvant chemotherapy (NCT) on the survival of patients with esophageal cancer. Methods: Clinical data of 275 cases of thoracic esophageal squamous cell carcinoma treated with neoadjuvant therapy combined with surgery from December 2011 to December 2015 were analyzed retrospectively. The data of treatment and follow-up were complete and analyzable. There were 70 cases in the NCRT group and 205 cases in the NCT group. The survival rate was calculated by Kaplan-Meier method and statistically compared by log-rank test, and multivariate analysis was performed by Cox regression model. Results: The median follow-up time was 32(3-84) months. The median survival time and recurrence-free survival time was 42(3-84) months and 30(3-84) months, respectively. The overall 3-and 5-year survival rates were 56.8% and 45.9%, respectively, and the 3-and 5-year recurrence-free survival rates were 45.1% and 38.9%, respectively. The median survival time in the NCRT and NCT groups was 46(7-84) and 40(4-74) months, and the median recurrence-free survival time was 31(3-84) and 28(3-69) months, respectively. The 3-and 5-year overall survival of the two groups were 59.1%, 47.1% and 56.3%, 47.5%(P=0.515), and the 3-and 5-year recurrence-free survival were 44.5%, 40.1% and 47%, 39%, respectively. There was no significant difference in the survival between two neoadjuvant therapy modes (P=0.554). Multivariate analysis showed that postoperative pathological TNM staging was an independent factor affecting the prognosis of patients with esophageal cancer (P=0.001). Conclusions The survival results of NCRT are similar to those of NCT. Postoperative pathological staging is an independent survival factor.

Objective To investigate the effect and mechanism of long noncoding RNA (lncRNA) PTEN pseudogene 1 (PTENP1) on the proliferation and migration of hepatocellular carcinoma (HCC) cells. Methods Lentiviral vectors expressing PTENP1 were constructed. HCC cells BEL-7404 were infected with LV003-GFP-PTENP1 and control vectors LV003-GFP. BEL-7404 cells stably expressing PTENP1 were constructed and the experimental and control groups were established. The proliferation and clone formation abilities of HCC cells in two groups were detected by CCK-8 assay and clonogenic assay. The migration ability of HCC cells was detected by wound healing assay. The expression of p44/42 mitogen-activated protein kinase (MAPK) and p38 MAPK proteins were detected by Western blot. Results The absorbance values A450 of the cells at 48 and 72 h in the experimental group were 1.4±0.3 and 2.3±1.1, signiifcantly lower compared with 3.2±1.7 and 3.4±1.1 in the control group (t=-5.78,-4.23;P<0.05). The number of cell clone formation in the experimental group was 55±12, signiifcantly less than 154±45 in the control group (t=-3.98, P<0.05). The percentage of cell migration in the experimental group was (21.7±2.6)%, signiifcantly lower than (57.7±4.9)%in the control group (t=-8.34, P<0.05). Western blot revealed that the expression of p44/42 MAPK and p38 MAPK proteins in the experimental group was significantly down-regulated compared with those in the control group. Conclusion lncRNA PTENP1 can inhibit the proliferation and migration of HCC cells probably through regulating MAPK signaling pathway.

Objective To analyze the effect of microRNA-22-3p(miR-22-3p)of extracellular vesi-cles derived from bone marrow mesenchymal stem cells on premature ovarian failure.Methods Follicular fluids were provided by premature ovarian failure patients with in vitro fertilization or the second-gen-eration in vitro fertilization treatment and normal volunteers with the same treatment for infertility due to male factors;the exosomes derived from bone marrow mesenchymal stem cells were isolated by dif-ferential centrifugation;the morphology,particle size and marker proteins of isolated exosomes were analyzed by transmission electron microscopy,NanoSight LM10 analyzer and Western blotting.Granulosa cells were treated with cyclophosphamide(CTX),exosomes,miR-22-3 mimics and corresponding controls,and were divided into CTX group,control group,exosome incubation group,PBS treat-ment group,CTX+miR-22-3p group,CTX+miR-NC group,CTX+Exosomes group,and CTX+PBS group;gene expression was detected by Western blotting and quantitative real-time polymerase chain reaction(qRT-PCR).The 3-(4,5-Dimethylthazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)reagent and flow cytometry were used to detect cell viability and apoptosis.Results The extracted exosomes had typical goblet vesicles,the exosome marker proteins C cluster of differentia-tion 63(CD63)and C cluster of differentiation 9(CD9)were expressed in the extracted exosomes,and exosome particle size was 80 to 150 nm;miR-22-3p was significantly lowly expressed in patients with premature ovarian failure and ovarian granulosa cells induced by CTX(P＜0.05),but was significantly highly expressed in ovarian granulosa cells incubated with exosomes derived from bone marrow mesenchymal stem cells(P＜0.05);the activity of granulosa cells in the CTX group was significantly lower than that in the control group,but was significantly higher in the CTX+miR-22-3p group or CTX+Exosomes group than that in the control group(P＜0.05);the apoptosis rate of granulosa cells in the CTX group was significantly higher than that in the control group,but was sig-nificantly lower in the CTX+miR-22-3p or CTX+Exosomes group than that in the control group(P＜0.05).Conclusion The miR-22-3p of extracellular vesicles derived from bone marrow mes-enchymal stem cells can inhibit ovarian granulosa cell injury induced by CTX.

Objective To analyze the effect of microRNA-22-3p(miR-22-3p)of extracellular vesi-cles derived from bone marrow mesenchymal stem cells on premature ovarian failure.Methods Follicular fluids were provided by premature ovarian failure patients with in vitro fertilization or the second-gen-eration in vitro fertilization treatment and normal volunteers with the same treatment for infertility due to male factors;the exosomes derived from bone marrow mesenchymal stem cells were isolated by dif-ferential centrifugation;the morphology,particle size and marker proteins of isolated exosomes were analyzed by transmission electron microscopy,NanoSight LM10 analyzer and Western blotting.Granulosa cells were treated with cyclophosphamide(CTX),exosomes,miR-22-3 mimics and corresponding controls,and were divided into CTX group,control group,exosome incubation group,PBS treat-ment group,CTX+miR-22-3p group,CTX+miR-NC group,CTX+Exosomes group,and CTX+PBS group;gene expression was detected by Western blotting and quantitative real-time polymerase chain reaction(qRT-PCR).The 3-(4,5-Dimethylthazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)reagent and flow cytometry were used to detect cell viability and apoptosis.Results The extracted exosomes had typical goblet vesicles,the exosome marker proteins C cluster of differentia-tion 63(CD63)and C cluster of differentiation 9(CD9)were expressed in the extracted exosomes,and exosome particle size was 80 to 150 nm;miR-22-3p was significantly lowly expressed in patients with premature ovarian failure and ovarian granulosa cells induced by CTX(P＜0.05),but was significantly highly expressed in ovarian granulosa cells incubated with exosomes derived from bone marrow mesenchymal stem cells(P＜0.05);the activity of granulosa cells in the CTX group was significantly lower than that in the control group,but was significantly higher in the CTX+miR-22-3p group or CTX+Exosomes group than that in the control group(P＜0.05);the apoptosis rate of granulosa cells in the CTX group was significantly higher than that in the control group,but was sig-nificantly lower in the CTX+miR-22-3p or CTX+Exosomes group than that in the control group(P＜0.05).Conclusion The miR-22-3p of extracellular vesicles derived from bone marrow mes-enchymal stem cells can inhibit ovarian granulosa cell injury induced by CTX.