ABSTRACT
Objective:To investigate the role and mechanism of pancreatic stellate cells (PSCs) and pancreatic cancer cells (PCCs) in the angiogenesis of pancreatic cancer.Methods:The experimental study was conducted. The human PSCs and PCCs and human umbilical vein endothelial cells (HUVECs) were cultured in vitro. HUVECs was treated with PSCs/PCCs supernatants and matrix metalloproteinase (MMP) inhibitor of different types and concentrations. As controls, HUVECs treated with complete endoprime medium (C/E) and DMEM/Ham's F12 medium (D/F) were set as the C/E group and the D/F group, respectively. Observation indicators: (1) proliferation of HUVECs under different conditions; (2) tube formation of HUVECs under different conditions; (3) migration of HUVECs under different conditions; (4) expression of MMP-2 in the supernatants of PSCs and PCCs; (5) effect of MMP inhibitor GM6001 on migration of HUVECs. Measurement data with normal distribution were represented as Mean± SD, comparison among groups was conducted using the one way ANOVA and comparison between groups was conducted using the LSD- t test. Results:(1) Proliferation of HUVECs under different conditions. Results of HUVECs proliferation assay using 5-ethynyl-2′-deoxyuridine (EdU) labeling showed that the binding rate of EdU in the HUVECs of D/F group and HUVECs treated with supernatants of different concentration (25%, 50%, 75%, 95%) of PSCs was 12.4%±1.0%, 24.5%±2.9%, 25.3%±3.0%, 22.8%±2.0%, 22.9%±2.8%, respectively, showing a significant difference among them ( F=8.60, P<0.05). There were significant differences in the binding rate of EdU between HUVECs in the D/F group and HUVECs treated with supernatants of different concentration (25%, 50%, 75%, 95%) of PSCs, respectively ( P<0.05). The binding rate of EdU between HUVECs in the D/F group and HUVECs treated with supernatants of different concentration (25%, 50%, 75%, 95%) of PCCs was 12.4%±1.0%, 30.0%±3.2%, 32.1%±1.0%, 32.3%±3.5%, 26.2%±5.6%, respectively, showing a significant difference among them ( F=11.93, P<0.05). There were significant differences in the binding rate of EdU between HUVECs in the D/F group and HUVECs treated with supernatants of different concentration (25%, 50%, 75%, 95%) of PSCs, respectively ( P<0.05). (2) Tube formation of HUVECs under different conditions. Number of tube formation, length of tube in the HUVECs of D/F group and HUVECs treated with PSCs supernatants was 15.2±2.3, (12.1±1.5)mm and 49.7±3.2, (39.8±2.3)mm, respectively, showing significant differences between the two groups of HUVECs ( P<0.05). (3) Migration of HUVECs under different conditions. Results of single cell tracing experiment showed that the migration rate of HUVECs treated with supernatants of different ratio of PSCs and PCCs was faster than that of HUVECs in the D/F group, and the enhancement effect of supernatants of PSCs and PCCs was dose-dependent. The migration rate of HUVECs treated with mix supernatants of different concentration of PSCs and PCCs and supernatants of co-cultured PSCs and PCCs was faster than that of HUVECs in the D/F group. The migration rate of HUVECs treated supernatants of co-cultured PSCs and PCCs was faster than that of HUVECs treated with mix supernatants of different concentration of PSCs and PCCs, showing a synergistic effect in the HUVECs treated supernatants of co-cultured PSCs and PCCs. (4) Expression of MMP-2 in the supernatants of PSCs and PCCs. Results of gelatine zymography showed that the MMP-2 expression levels decreased sequentially in super-natants of co-cultured PSCs and PCCs, supernatants of PSCs, mix supernatants of PSCs and PCCs and supernatants PCCs. (5) Effect of MMP inhibitor GM6001 on migration of HUVECs. Results of single cell tracing experiment showed that the migration rate of HUVECs treated with PSCs supernatants combined with different concentration of GM6001 (0, 1, 10, 25 μmol/L) was (25.70±2.06)μm/h, (18.37±1.61)μm/h, (16.20±0.26)μm/h, (15.99±0.58)μm/h, respectively, showing a significant difference among them ( F=11.39, P<0.05). There were significant differences in the migration rate between HUVECs treated with PSCs supernatants combined with 1, 10, 25 μmol/L GM6001 and HUVECs treated with PSCs supernatants ( P<0.05). The migration rate of HUVECs treated with mix super-natants of PSCs and PCCs combined with different concentration of GM6001 (0, 1, 10, 25 μmol/L) was (30.06±3.70)μm/h, (22.76±1.56)μm/h, (23.87±2.84)μm/h, (22.10±2.35)μm/h, respectively, showing a significant difference among them ( F=4.06, P<0.05). There were significant differences in the migration rate between HUVECs treated with mix supernatants of PSCs and PCCs combined with 1, 10, 25 μmol/L GM6001 and HUVECs treated with mix supernatants of PSCs and PCCs ( P<0.05). Conclusions:Both PSCs and PCCs can promote the proliferation, migration and angiogenesis of HUVECs in vitro experiment. Releasing of MMP-2 by interaction between PSCs and PCCs is an important factor to stimulate endothelial cell migration, which increases the stimulating activity of angiogenesis, especially the migration ability of HUVECs.
ABSTRACT
Objective To investigate the features of time distribution in the onset of aortic dissection (AD).Methods The retrospective cross-sectional study was conducted.The clinical data of 476 AD patients who were admitted to the First Affiliated Hospital of Anhui Medical University from January 2009 to June 2017 were collected.The patients were divided by the following criteria:gender,age [youth(＜45 years),middle-age (45-59 years) and elderly (≥ 60 years)],Stanford types (type A and type B),with or without hypertension.All variables were analyzed by circular distribution statistics to illuminate the features of time distribution in the onset of AD (monthly rhythm and circadian rhythm).Observation indicators:(1) overall time distribution of AD;(2) time distribution of subgroups with different genders;(3) time distribution of subgroups with different age;(4) time distribution of subgroups with different Stanford types;(5) time distribution of subgroups with or without hypertension.Measurement data with normal distribution were represented as (x)±s and count data were described as constituent ratio.The circular distrbution statistics were used to calculate time data of onset after trigonometric function transformation.The monthly rhythm and circadian rhythm were done using the Rayleigh test (Z value).Results (1) Overall time distribution of AD:the AD patients had the monthly rhythm and circadian rhythm (Z=14.79,31.60,P＜0.05).The months with the maximum and minimum cases were November (59 cases) and August (24 cases) respectively,the peak day was on January 12.AD often occurred from 16:00 to 17:00 (37 cases) but barely occurred from 3:00 to 4:00 (8 cases),with a peak of 14:55.(2) Time distribution of subgroups with different gender:male subgroup had the monthly rhythm and circadian rhythm (Z =11.28,27.81,P＜0.05);female subgroup had the monthly rhythm and circadian rhythm (Z=3.48,4.37,P＜0.05).(3) Time distribution of subgroups with different age:patients in the youth subgroup had no monthly rhythm (Z=1.33,P＞0.05),and there was the circadian rhythm (Z=4.29,P＜0.05);patients in the middle-age subgroup had the monthly rhythm and circadian rhythm (Z =7.48,17.41,P＜0.05);patients in the old-age subgroup had the monthly rhythm and circadian rhythm (Z =6.62,11.04,P ＜ 0.05).(4) Time distribution of subgroups with different Stanford type:patients inthe type A subgroup had no monthly rhythm (Z=1.60,P＞0.05),and there was the circadian rhythm (Z=10.51,P＜0.05);patients in the type B subgroup had the monthly rhythm and circadian rhythm (Z=13.94,21.70,P＜0.05).(5) Time distribution of subgroups with or without hypertension:subgroups with hypertension had the monthly rhythm and circadian rhythm (Z =12.08,29.81,P＜ 0.05).Subgroups without hypertension had no monthly rhythm (Z=3.84,P＞0.05),showing a statistically significant difference in the circadian rhythm (Z=4.78,P＜0.05).Conclusion AD often occurs in cold months and afternoon.
ABSTRACT
Objective To explore clinical implications of pleural effusion in thoracic endovascular aorta repair (TEVAR) of type B aortic dissection.Methods Clinical data of 28 patients (23 males,5 females) hospitalized from Jan 2015 to Dec 2016 were analyzed retrospectively.There were ruptured aortic dissection (RAD) (n =7) and the contained aortic dissection (CAD) (n =21).26 patients underwent TEVAR,and two patients received conservative treatment.Results 26 patients received TEVAR and operations were successful.2 patients treated conservatively died.Six patients had bilateral pleural effusion,while 20 had left pleural effusion and two had right pleural effusion.The distribution of pleural effusion was significantly different between CAD and RAD group (x2 =10.4,P ＜ 0.05),and the rupture risk was the highest in right sided pleural effusion.The median volume of pleural effusion on right side in RAD group are higher than that in CAD group (Z =-3.293,P =0.001).One patient died of sudden death on post-op 9th day.Pleural effusion disappeared in all 24 patients who were followed-up for more than 3 months.There were no ensuing pleural thickening,pulmonary atelectasis,and lung consolidation.Conclusious Pleural effusion on left side are common in type B aortic dissection,while bulk right pleural effusion may indicate impending rupture.Endovascular therapy is a feasible,safe and effective therapy for aortic dissection with pleural effusion.
ABSTRACT
Pancreatic cancer is still a dismal disease.Angiogenesis is very important for the development of pancreatic cancer.Pancreatic stellate cells (PSCs) are the main source of extra-cellular matrix of pancreatic cancer and they provide advantageous microenvironment for cancer cells.PSC could promote the angiogenesis of pancreatic cancer both in vitro and in vivo.Further studies of the angiogenesis of pancreatic cancer are helpful in learning the characteristics of development and metastasis of pancreatic cancer,and provide new treatment method in the cellular and molecular levels.
ABSTRACT
Objective To detect aberrant methylation in the promoter of FHIT and RASSF1A genes in peripheral plasma and tumor tissues from patients with hepatocellular carcinoma (HCC) and to determine its clinical significance.Methods The methylation status of FHIT and RASSF1A genes in peripheral plasma and tumor tissues from 36 patients with HCC were detected by methylation-specific polymerase chain reaction(MSP).The correlation between methylation status in plasma and clinicopathological features was analyzed.Results The frequency of promoter methylation of FHIT in tissues was 75％ (27/36) and in plasma 52.8％ (19/36),and the correlation coefficient was r=0.482 (P=0.003).The frequency of promoter methylation of RASSF1A in tumor tissues was 83.3％ (30/36) and in plasma 61.1％ (22/36),and the correlation coefficient was r=0.561 (P=0.0004).Aberrant methylation of FHIT,RASSF1A gene in the plasma and tissues had no correlation with the patients' clinicopathological features such as gender,age,HBV/HCV infection,hepatic cirrhosis,tumor size,alpha-fetoprotein (AFP) level,pathological grade,staging,vascular tumour thrombus and recurrence.The sensitivity of AFP ≥400 μg/L was 44.4％,and AFP ≥20 μg/L 69.4％.The sensitivity of FHIT and RASSF1A gene promoter hypermethylation in 36 HCC patients was 72.2％.In 20 patients whose AFP ＜400 μg/L,the frequency of hypermethylation of the two genes together was 80％.When AFP ＜20 μg/L,the frequency of hypermethylation of the two genes together was 54.5 ％.Conclusions There was a significant concordance between plasma and tumor tissue methylation profiles.The methylation status in plasma and tumor tissues had no correlation with the patients' clinicopathological features.Combining promoter methylation of FHIT and RASSF1A genes was superior to AFP in the diagnosis of HCC.
ABSTRACT
Objective To sum up our experience on the diagnosis and treatment of rare hepatic tumors. Methods The data of 25 patients with rare liver tumors admitted in our hospital from May 2005 to January 2010 were analyzed retrospectively. Results The final pathologic diagnosis of focal nodular hyperplasia was made in 6 cases, and the diagnosis of vascular leiomyoma, hilar neurilemoma, intrahepatic aneurysm, biliary cystadenoma, hepatic hamartoma, biliary villous adenoma, and hepatic diffuse large B-cell lymphoma was established in one each case, respectively. The diagnosis of angiomyolipoma in 2patients, primary liver gastroimestinal stromal tumor in 2 patients, hepatoblastoma in 5 patients and liver undifferentiated sarcoma in 3 patients was established. Preoperative ultrasonography, CT and MRI were performed in 24, 22 and 6 patients respectively. Preoperative tentative diagnosis was finally confirmed by pathology in only 3 (16.7%) cases, all by CT report. Preoperative diagnosis was consistent with postoperative pathology in 5 patients (20%); All patients underwent liver resection including hemihepatectomy in 7 patients, hepatic lobectomy in 7 patients, segmentectomy in 9 patients and tumor enucleation in 2 patients; There was no recurrence after resection of benign, low malignant tumors and hepatic diffuse large B-cell lymphoma; Postoperative follow-up was made for all the 5 cases of malignant tumours, and there was recurrence in 3 cases. These 3 eases underwent second resection and there were no recurrences after reoperation. The two recurrent patients died with a mean survival of 4 months.Conclusions The preoperative correct imaging diagnostic rate for rare hepatic tumors is low. Surgery is the most effective therapy and reoperation should always be attempted for tumor recurrence in order to prolong survival.