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Objective To investigate the effect of targeted silencing Notch1 on proliferation and apoptosis of human non-small cell lung cancer stem cells.Methods Lung cancer A549 cells and SPC-A-1 cells were selected and divided into control group,Nc-shRNA group and Notch1-shRNA group.The Nc-shRNA group was a negative control RNAi lentivirus group,and the Notch1-shRNA group was a Notch1 inhibitory RNAi lentivirus group.The lentiviral-mediated shRNA interference technology was used to target the silencing of Notch1.The silencing effect of Notch1 gene was verified by quantitative real time polymerase chain reaction (qRT-PCR) and Western blotting.Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) and sarcosphere formation assay.Apoptosis was detected by Annexin V/7-AAD double staining.Western blotting was used to detect the expression of proliferating cell nuclear antigen (PCNA),B-cell lymphoma-2 (Bcl-2) and Notch1 downstream gene Hes-1.Results The results of qRT-PCR showed that the relative expression levels of Notch1 in control group,Nc-shRNA group and Notch1-shRNA group in A549 cells and SPC-A-1 cells were 1.000 ± 0.000,0.937 ± 0.025,0.490 ± 0.036 and 1.000 ± 0.000,1.077 ± 0.070,0.373± 0.038,with statistically significant differences (F =359.707,P ＜0.001;F =210.455,P ＜0.001),further paired comparison,the relative expression of Notch1 in Notch1-shRNA group was significantly lower than that in Nc-shRNA group (all P ＜ 0.05).Western blotting showed that the expressions of Notch1 protein in A549 cells and SPC-A-1 cells were consistent with the mRNA results.MTT assay showed that the 24 h A values of A549 cells in control group,Nc-shRNA group and Notch1-shRNA group were 0.209 ± 0.005,0.219 ± 0.009,0.159 ±0.006,48 h A values were 0.293 ± 0.004,0.302 ± 0.004,0.205 ± 0.005,72 h A values were 0.450 ± 0.003,0.430 ± 0.012,0.348 ± 0.017,with statistically significant differences (F =79.487,P＜0.001;F =508.664,P ＜0.001;F =57.156,P ＜0.001),further paired comparison,the proliferation ability of Notch1-shRNA group was significantly lower than that of Nc-shRNA group at 24,48,72 h (all P ＜ 0.05).The 48 h A values of SPC-A-1 cells in control group,Nc-shRNA group and Notch1-shRNA group were 0.438 ±0.022,0.412 ± 0.015,0.364 ± 0.010,72h A values were 0.540 ± 0.016,0.519 ± 0.009,0.438 ± 0.019,with statistically significant differences (F =15.667,P =0.004;F =37.299,P ＜ 0.001),further paired comparison,the proliferation ability of Notch1-shRNA group was significantly lower than that of Nc-shRNA group at 48 h and 72 h (all P ＜ 0.05).The sphere sizes of control group,Nc-shRNA group and Notch1-shRNA group in A549 cells were (149.667 ± 6.506) μm,(136.667 ± 7.095) μm,(86.676 ± 7.638) μm,with statistically significant difference (F =65.940,P ＜ 0.001).The sphere sizes of the three groups in SPC-A-1 cells were (118.667 ± 6.658) μm,(128.000 ± 7.000) μm,(60.675 ± 4.509) μm,with statistically significant difference (F =105.372,P ＜0.001).Further paired comparison,the sphere size of Notch1shRNA group was significandy smaller than that of Nc-shRNA group in the two kinds of cells (all P ＜ 0.05).The apoptosis rates of control group,Nc-shRNA group and Notch1-shRNA group in A549 cells and SPC-A-1cells were (0.489 ± 0.014)％,(0.633 ± 0.021)％,(1.683 ± 0.221)％ and (1.323 ± 0.194)％,(1.690 ± 0.188) ％,(3.017 ± 0.356) ％,with statistically significant differences (F =77.660,P ＜ 0.001;F=32.200,P =0.001),further paired comparison,the apoptosis rate of Notch1-shRNA group was significantly higher than that of Nc-shRNA group in the two kinds of cells (all P ＜ 0.05).Western blotting showed that the expressions of PCNA,Bcl-2 and Hes-1 in control group,Nc-shRNA group and Notch1-shRNA group in A549 cells were statistically significant (F =155.343,P ＜ 0.001;F =22.576,P =0.002;F =70.108,P＜0.001),and the expressions of PCNA,Bcl-2 and Hes-1 in the three groups in SPC-A-1 cells were statistically significant (F =49.419,P ＜0.001;F =28.090,P =0.001;F =12.040,P =0.007).Further paired comparison,the expressions of PCNA,Bcl-2 and Hes-1 in Notch1-shRNA group were significantly lower than those in Nc-shRNA group in the two kinds of cells,and the differences were statistically significant (all P ＜0.05).Conclusion Targeted silencing of Notch1 can reduce the proliferation activity of lung cancer stem cells and promote apoptosis,which may be related to the down-regulation of its downstream gene Hes-1.
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Objective@#To investigate the principles of diagnosis and treatment of breast cancer during pregnancy.@*Methods@#Clinical data of patients with breast cancer during pregnancy admitted to Obstetrics and Gynecology Hospital of Fudan University between January 2012 to July 2017 were analyzed retrospectively. A total of 17 patients were diagnosed with breast cancer in pregnancy, the median age was 32 years (range from 25 to 45 years old), pathological staging revealed 2 patient with stage 0, 1 with stage Ⅱa, 7 with stage Ⅱb, 1 with stage Ⅲa, 2 with stage Ⅲc, 4 with stage Ⅳ.@*Results@#Thirteen patients received surgical treatment in pregnancy, the gestational age at surgery was (27.7±4.6) weeks; 2 patients with ductal carcinoma in situ received mastectomy, 11 patients with breast cancer underwent modified radical mastectomy. In patients undergoing surgery during pregnancy, no prophylactic contractions were used in 4 patients who had been treated earlier, there were 2 patients with frequent contractions within 24 hours after operation in these patients. Follow-up 9 patients were given oral nifedipine to prevent contractions, no obvious contractions occurred after the operation. Seven patients received chemotherapy during pregnancy; the chemotherapy of 4 cases of triple negative breast cancer was weekly paclitaxel sequential epirubicin and cyclophosphamide, the chemotherapy of the other three patients was docetaxel sequential epirubicin and cyclophosphamide. Fifteen patients underwent cesarean section to terminate pregnancy, 2 patients underwent spontaneous labor. The gestational age of birth was (36.9 ±1.3) weeks. Less than 35 weeks of termination of pregnancy occurred in one patient, the fetus was delivered to the neonatal intensive care unit due to neonatal respiratory distress syndrome, and suffered from congenital dysaudia. The prognosis of the other 16 survived infants was good. The median follow-up time was 10 months (range from 4 to 27) months, in 13 patients of stage 0 to Ⅲc, one patient were diagnosed with bone metastasis at 12 months after surgery, the remaining 12 patients had no disease progression, the progression free survival rate was 12/13, the overall survival rate was 13/13. Among the 4 patients with stage Ⅳ, one died in 7 months after delivery, one had new liver metastasis in 8 months after delivery. The remaining 2 patients were in stable condition.@*Conclusions@#Breast cancer in pregnancy can be treated effectively, multidisciplinary cooperation and detailed assessment of maternal-fetal risks and benefits are necessary. Chemotherapy during pregnancy is safe for maternal-fetal, but it needed a large sample of clinical studies and long-term follow-up. The neonatal outcome was associated with gestational age, and therefore premature delivery was avoided as much as possible during treatment.
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Notch signaling pathway is involved in the abnormal differentiation and self-renewal of lung cancer stem cells.The further studies for the roles of Notch signaling pathway in the regulation of lung cancer stem cells are expected to find new targets in the diagnosis and treatment of lung cancer.Inhibitors of the Notch signaling pathway may be effective in the treatment of lung cancer.Lung cancer stem cells are thought to be a major cause of recurrence of lung cancer,therefore,targeted therapy for lung cancer stem cells may be more effective than treatment for the entire tumor.
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Objective@#To explore the impact of weight management and related medication intervention based on body weight changes on cardiac function among patients with chronic congestive heart failure (CHF).@*Methods@#Using prospective, randomized, controlled study methods, consecutive CHF patients, who hospitalized in our department from June 2014 to June 2016 (n=350), were randomly divided into intervention group (n=175) and control group (n=175). Patients in the intervention group received weight management guidance and the post discharge diuretic drugs regimen was adjusted based on body weight changes. The control group received routine medical care post discharge. Left ventricular ejection fraction (LVEF), B type natriuretic peptide precursor (NT-proBNP), 6 minutes walk distance and NYHA classification at one day before discharge and after 6 months were compared between the two groups respectively.@*Results@#Follow-up visit data were not available from 6 patients in the control and intervention group respectively. NYHA classification, LVEF, NT-proBNP and 6 minutes walk distance were similar between the two groups at one day before discharge (all P>0.05). After 6 months, the LVEF and 6 minutes walk distance were significantly higher while NT-proBNP level was significantly lower in the intervention group compared to the control group (all P<0.01). Meanwhile, the LVEF and 6 minutes walk distance were significantly increased, while NT-proBNP was significantly reduced at 6 months post discharge compared to one day before discharge in the intervention group (all P<0.01). The LVEF was also significantly improved (P=0.035), but the NT-proBNP and 6 minutes walk distance were similar (P were 0.328 and 0.807 respectively) at 6 months after discharge compared to one day before discharge in the control group. The NYHA classification was significantly lower in intervention group and in control group at 6 months after discharge compared to one day before discharge (Z=5.154, P<0.01 and Z=10.497, P<0.01), and the NYHA classification improved more in the intervention group than in control group at 6 months after discharge (Z=9.235, P<0.01). The re-hospitalization rate of CHF patients in intervention group was 11.83% (20/169), which was significantly lower than the control group (33.14% (56/169), χ2=21.99, P<0.01). At 6 months follow up, body weight remained unchanged in the intervention group, while body weight tended to be higher in the control group compared to one day before discharge.@*Conclusion@#The weight management and diuretic drug regimen adjudgment intervention based on body weight changes can improve cardiac function and reduced re-hospitalization rate in CHF patients.
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The stress gastric ulcer is due to a stress-induced gastric erosions, superficial ulcers and bleeding gastric mucosal le-sions characterized by stress. Its etiology is closely associated with lives, the incidence is increasing. In recent years, the the basic re-search of drugs focuses on inhibition of oxidative damage caused by stress, anti-inflammatory cytokine expression, and regulation of gastric acid, etc., and made a lot of success. Treatment choice of stress ulcer drug gradually increased. The effects of plant extracts on stress ul-cer get more and more attention. This article reviews research progress of the treatment drugs of stress ulcer in the current stage.
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Aim To investigate the alteration of Wnt/β-catenin signaling pathway in early diabetic rat myo-cardium and clarify its role in development of diabetic cardiomyopathy. Methods The diabetes mellitus ( DM) model was prepared by intraperitoneal injection of streptozotocin ( STZ, 60 μg · g-1 ) . The alteration of Wnt/β-catenin signaling pathway was determined by Western blot and immunohistochemistry. HE staining was used to analyze the change of myocardial pathologi-cal structure. Results Cardiac histological analyses revealed that DM induced cardiomyocyte degeneration and necrosis. Myocardial Wnt2, β-catenin and c-Myc were enhanced in 2 wk DM compared with control group while DKK1 showed no significant alteration. Conclusion Wnt/β-catenin signaling pathway is acti-vated in early diabetic myocardial injury. Further re-searches on its role in DM myocardium may find a new therapeutic target for diabetic cardiomyopathy.
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BacKground:Acetic acid spray combined with narrow-band imaging ( NBI ) endoscopy can improve the earIy diagnosis of coIorectaI cancer and precancerous Iesions. Aims:To assess the vaIue of acetic acid spray combined with NBI in diagnosis of earIy coIorectaI cancer and precancerous Iesions. Methods:A totaI of 236 coIorectaI cancer and precancerous Iesions in 180 patients detected by screening endoscopy from Nov. 2011 to JuIy 2013 were enroIIed in this study. Acetic acid spray combined with NBI( combination group),singIe NBI,and indigo carmine staining were used to observe pit pattern,bIood capiIIary forms of the Iesion and mucosa whitening time,and vaIues of the three methods in diagnosis of earIy coIorectaI cancer and precancerous Iesions were evaIuated. Results:Image cIarity of pit pattern in combination group was significantIy improved than that in NBI group and indigo carmine group(χ2 =15. 113,P=0. 000;χ2 =6. 128,P=0. 013);image cIarity of bIood capiIIary forms in combination group was significantIy improved than that in NBI group(χ2 =7. 384,P =0. 007). The overaII coincidence rates of diagnosis in combination group,NBI group, indigo carmine group were 91. 9%,89. 0%,86. 9%,respectiveIy. The diagnosis rate of pit pattern in combination group was significantIy higher than that in NBI group(χ2 =4. 490,P=0. 034),but no significant difference in bIood capiIIary forms was found between combination group and NBI group. The sensitivity of type Ⅲ,Ⅳ,Ⅴ pit pattern,type Ⅱ,ⅢbIood capiIIary forms,mucosa whitening time <30 seconds for diagnosis of coIorectaI cancer and precancerous Iesions in combination group were 94. 0%,92. 8%,92. 2%,respectiveIy,the specificity were 87. 4%,88. 1%,88. 0%, respectiveIy,and the accuracy were 91. 5%,91. 1%,90. 7%,respectiveIy. The sensitivity,specificity and accuracy of type ⅤN or highIy irreguIar ⅤI pit pattern,type ⅢB bIood capiIIary forms and mucosa whitening time 0-5 seconds for predicting invasive carcinoma in combination group were aII quite weII. Conclusions:Acetic acid spray combined with NBI can cIearIy show the pit pattern and bIood capiIIary forms of earIy coIorectaI cancer and precancerous Iesions,and is fairIy consistent with the pathoIogicaI diagnosis. It is heIpfuI for diagnosing the nature of Iesion and depth of invasion.
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Objective To evaluate the effect of ultrasound-guided vacuum-assisted minimal invasive resection(Mammotome procedure)of breast lumps through the retromammary space. Methods Seven hundred and eighty-seven patients in Obstetrics and Gynecology Hospital Affiliated to Fudan University from Jan. 2011 to May 2012 were underwent ultrasound-guided Mammotome operation through the retromammary space (retromammary space group,385 cases),or adjacent the lumps,and followed by post-operation visits regularly (Mammotome operation adjacent the lumps group,402 cases). The operation effects were compared between the two groups. Results All cases were followed up for 12 months. The period of Mmmotome operation through the retromam mary space and the rate of resection were(48 ± 6)min and(52 ± 4)min,99. 48%(383 / 385), 99. 25%(399 / 402),perspectively,in group of Mammotome operation through the retromammary space and Mammotome operation adjacent lumpsand. There was no significant difference between the two groups( P> 0. 05). The amount of procedural bleeding,the incidence of ecchymosis,local hematoma and the number of incision in group of Mammotome operation through the retromammary space were(8 ± 3)ml,2. 34%(9 / 385), 0. 52%(2 / 385),(1. 3 ± 0. 6)respectively,which were significantly lower than those in Mammotome operation adjacent the lumps group((14 ± 6)ml,8. 71%(35 / 402),2. 74%(11 / 402),(2. 4 ± 0. 3)respectively). There were statistical difference between two groups( P = 0. 003,P < 0. 001,P = 0. 001,P = 0. 006). The rate of satisfaction in group of Mammotome operation through the retro-mammary space was 98. 70%(380 / 385),which is significantly higher than in group of Mammotome operation adjacent the lumps(89. 30%(359 / 402),P< 0. 01). Conclusion The therapy approach of ultrasound-guided Mammotome operation through the retromammary space has lower hemorrhagic complication,as well as the better effect with special advantages. Therefore it has prospective clinical application.
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Objective:To investigate the impact of polysomy 17 of breast cancer on testing results of human epidermal growth factor receptor 2 (HER2) and its clinicopathologic significance. Methods:Seventy-one patients with primary invasive breast carcinoma were studied. The HER2 gene and chromosome 17 copy numbers were determined by dual-color fluorescence in situ hybridization (FISH). The testing results were expressed by absolute HER2 gene copy number or the ratio of HER2 to chromosome 17. Based on the FISH testing results and HER2 protein expression determined by immunohistochemistry the results were compared between different groups divided by related clinicopathologic parameters.Results:All patients who had doubtable FISH results, either by absolute HER2 copy number (14 of 71 patients; 19.7%) or by the ratio HER2/chromosome 17 (2 of 71 patients, 2.8%), displayed polysomy 17. Polysomy 17-positive patients had no significant difference with HER2-negative patients in tumor grade, lymph node metastasis, and estrogen receptor (ER) expression (all P>0.05); but compared with HER2-positive patients, they showed lower tumor grade (50.0% vs 81.5%, P=0.025), higher rate of negative lymph node (55.6% vs 25.9%, P=0.045), and higher rate of ER positive expression (83.3% vs 41.7%, P=0.005) and progesterone receptor(PR)positive expression (87.5% vs 44.4%, P=0.003).Conclusion:Compared with HER2 gene amplification group, polysomy 17-positive group tends to have negative HER2 gene expression. Polysomy 17 influences the testing results of HER2 and may be the main factor that caused doubtable results in FISH examination.
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BACKGROUND: The small change of the membrane potential (Em) of vascular smooth muscle cells (VSMC) could result in significant alteration in tension, and the membrane depolarization was closely associated with the development of hypertension.OBJECTIVE: To study the resting Em of VSMC from the middle cerebral artery (MCA) of stroke-prone spontaneously hypertensive rats (SHRSP) and the reactivity of Em to KCl and norepinephrine (NE).DESIGN: An animal experiment of comparative analysis.SETTING: Institute of Materia Medica, Shandong Academy of Medical Sciences.MATERIALS: Eighteen male SHRSP and 26 male Wistar rats were used.METHODS: ① Rats in both groups were anesthetized with intraperitoneal injection of ethyl carbamate (1 g/kg), the brains were removed quickly, and the Em of VSMC was recorded by intracellular microelectrodes. ② The effects of different concentrations of KC1 (10, 20, 50 mmol/L) and NE (10-7,10-6, 10-5 mol/L) on the Em of VSMC were observed.MAIN OUTCOME MEASURES: ① Em of VSMC in both groups; ②Effects of different concentrations of KC1 and NE on the Em of VSMC.RESULTS: ① The Em was obviously lower in SHRSP than in the Wistar rats [(-48.2±3.1), (-64.4±4.3) mV]. ② Both KC1 (10, 20, 50 mmol/L)and NE (10-7, 10-6, 10-5 mol/L) could cause the depolarization of Em of MCA in a concentration-dependent manner. The reactivity to MCA of SHRSP was obviously increased as compared with the Wistar rats.CONCLUSION: The resting Em of VSMC from the MCA of SHRSP was lower, and the reactivity of VSMC to KCl and NE was significantly stronger than that of Wistar rats.
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The effect of Sangge Mixture (SM) on cerebral circulation and metabolism in rabbits was observed.The results showed that SM increased the blood flow of brain and cerebral cortex in a dose-dependent manner and had no effect on the blood flow of femoral arteries. Meanwhile, SM markedly increase the oxygen consumption and glucose intake in rabbit brain. It is suggested that SM can improve cerebral circulation, with a certain degree of selectivity.