RESUMO
Objective:To investigate the synergistic effects and molecular mechanisms of dihydroartemisinin(DHA) and sorafenib(SOR) in inducing ferroptosis in anaplastic thyroid cancer(ATC) cells.Methods:CCK-8 and flow cytometry assays were performed to detect the effects of DHA and SOR on the proliferation and ferroptosis of ATC cells(CAL-62). Real-time fluorescence quantitative PCR and Western blotting assays were performed to detect the expressions of ferroptosis-related genes glutathione peroxidase 4(GPX4), solute carrier family 7 member 11 gene(SCL7A11), lipoxygenase-15(LOX-15), and p53. The levels of iron death intermediate metabolites including lactate dehydrogenase(LDH), glutathione(GSH), malondialdehyde(MDA), ferrous ion(Fe 2+ ), nitric oxide(NO), and reactive oxygen species(ROS)were measured by corresponding assay kits. The corresponding inhibition of DHA and SOR on ATC in vivo was analyzed in a tumor model in nude mice. Results:Compared with the control group, DHA, SOR, and DHA+ SOR treatment significantly inhibited cell proliferation and apoptosis in a dose-dependent manner( P<0.001), with increased LDH, Fe 2+, MDA, and ROS contents and reduced GSH activity( P<0.001), which were promoted by ferrous sulfate(FeSO 4)and reversed by ferroptosis inhibitor-1. Compared with the control group and the drug monotherapy group, 15-LOX-2 and p53 expressions were upregulated in DHA+ SOR group while GPX4 and SCL7A11 expressions were decreased( P<0.001), without significant difference in 15-LOX-1 protein content. In addition, NO level was significantly increased in DHA+ SOR group( P<0.001). DHA and SOR inhibited tumor growth of ATC in vivo. Conclusion:DHA and SOR synergistically induced ferroptosis via upregulating the expression of 15-LOX-2 gene and inhibiting NO synthesis in ATC cells.
RESUMO
Objective:To study the effect of ranibizumab on retinal oxidative stress in a rat model of choroidal neovascularization (CNV) and its mechanism.Methods:Sixty SPF male SD rats aged 10 weeks were randomly divided into normal control group, model control group, ranibizumab group, nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor (ML385) group, ranibizumab+ ML385 group, with 12 rats in each group according to a random number table.Except for the normal control group, the CNV model was established in the other four groups via krypton laser induction.According to grouping, the ranibizumab group, ML385 group and ranibizumab+ ML385 group were intravitreally injected with 1 μl of ranibizumab, ML385 and ranibizumab+ ML385, respectively.Model control group and normal control group received an intravitreal injection of normal saline of equal volume.The CNV area was measured through choroidal wholemounts.Pathological change of the retina was observed by hematoxylin and eosin staining.Expressions of Nrf2, superoxide dismutase (SOD) and quinone oxidoreductase 1 (NQO1) were detected using Western blot and real-time PCR.The use and care of animals complied with laboratory animal welfare guidelines.The study protocol was approved by the Laboratory Animal Welfare and Ethics Committee of Tengzhou Central People's Hospital (No.JN.No20210214S1200430[121]).Results:CNV areas of the model control group, ML385 group and ranibizumab+ ML385 group were (23.01±1.52)×10 3, (30.23±2.01)×10 3 and (18.56±1.85)×10 3 μm 2, respectively, which were significantly higher than (12.35±1.22)×10 3 μm 2 of ranibizumab group (all at P<0.001). The CNV area of ranibizumab+ ML385 group was smaller than that of model control group and ML385 group, and the CNV area of ML385 group was larger than that of model control group, showing statistically significant differences (all at P<0.001). Hematoxylin and eosin staining showed that the structural damage of the retinal pigment epithelium-choroid-sclera complex was slighter in ranibizumab group than model control group, severer in ranibizumab+ ML385 group than ranibizumab group but slighter than model control group, severer in ML385 group than model control group.The mRNA and protein expressions of Nrf2, SOD and NQO1 of ranibizumab group were lower than those of normal control group but higher than those of model control group, ML385 group and ranibizumab+ ML385 group, and the differences were statistically significant (all at P<0.05). The mRNA and protein expressions of Nrf2, SOD and NQO1 were higher in ranibizumab+ ML385 group than model control group and ML385 group, showing statistically significant differences (all at P<0.05). Conclusions:Ranibizumab can inhibit the growth of CNV induced by krypton laser and reduce RPE damage caused by retinal oxidative stress.The mechanism is related to the activation of Nrf2/ARE pathway.
RESUMO
Objective:To investigate and evaluate the efficacy of HeartCon left ventricular assist device (LVAD) in the treatment of adult patients with end-stage heart failure (ESHF).Methods:A prospective and observational study was conducted. Patients with ESHF who underwent LVAD implantation in the department of cardiac surgery of Teda International Cardiovascular Hospital from September 2020 to August 2021 were selected. The left ventricular end-diastolic diameter (LVEDD), left ventricular ejection fraction (LVEF), New York Heart Association (NYHA) classification, N-terminal pro-B type natriuretic peptide (NT-proBNP), and six minute walk distance (6MWD) before operation and 90 days after operation were compared. The incidence of equipment failures and major adverse events within 90 days after operation were recorded.Results:A total of 20 patients with ESHF were included, with 15 males and 5 females. Patients' age ranged from 20 to 67 years old, with an average of (50.2±13.6) years old. The range of body weight was 49.8-106.1 kg, with an average of (67.9±15.5) kg, and the body surface area (BSA) was from 1.49 to 2.32 m 2, with an average of (17.6±0.22) m 2. The operation process of all the patients were successful. The length of hospital stay ranged from 33 to 90 days, and the average was 56.0 (42.8, 75.0) days. Complications within 90 days after operation as follows, 2 cases with pericardial tamponade (10%), 1 case with cerebral hemorrhage (5%), 1 case with mediastinum infection (5%), 3 cases with acute renal injury (AKI, 15%), 5 cases with gastrointestinal bleeding (25%). There were no mechanical failure of LVAD and hemolysis events, right ventricular failure (RVF), cerebral infarction and death occurred. Compared with preoperative, the LVEDD significantly decreased (mm: 67.50±13.98 vs. 77.40±9.73), LVEF significantly increased (%: 34.80±9.76 vs. 22.70±5.62), NT-proBNP significantly decreased (ng/L: 2 028.65±1 752.05 vs. 4 796.45±4 355.40), 6MWD significantly increased (m: 385.20±144.12 vs. 85.81±63.50) at 90 days after operation, and the differences were statistically significant (all P < 0.05). 18 cases (90%) of the 20 patients reached NYHA classification Ⅰ and 2 cases (10%) reached NYHA classification Ⅱ, which were significantly improved compared with those before surgery (all patients' NYHA classification were Ⅳ before surgery). Conclusion:HeartCon LVAD can effectively improve the life quality of patients with ESHF, which has been proved safe and effective in clinical trials, but its long-term effects and complications need further observation and study.
RESUMO
Cerebral small vessel disease (CSVD) is a common and slowly progressive cerebrovascular disease. Its pathological mechanism involves vascular endothelial dysfunction, blood-brain barrier destruction, neuronal apoptosis, glial cell activation, and inflammatory reaction. Neurovascular unit is the basic unit of brain structure and function, and its pathological changes are closely associated with many cerebrovascular diseases. At present, the damage mechanism of neurovascular unit in CSVD has been paid more and more attention. This article reviews the damage mechanism of neurovascular unit in CSVD.
RESUMO
Objective@#To evaluate the clinical value of 16-slice spiral CT low dose chest scanning in the diagnosis of pulmonary tuberculosis.@*Methods@#From June 13, 2014 to June 20, 2017, 80 patients with pulmonary tuberculosis in Yuyao People's Hospital were selected in the study.The conventional chest dose(control group) and low dose(observation group) of 16-slice spiral CT were used.The diagnostic accuracy, radiation dose of the two methods, lymph node or vascular space display and image features were observed.@*Results@#The diagnostic accuracy of the two methods had no statistically significant difference (P>0.05). The dose product length, the CT dose index in the observation group were (32.98±2.57) mGycm, (44.29±3.47), respectively, which were significantly lower than those in the control group[(127.66±5.03)mGycm, (44.29±3.47)](t=106.01, 21.05, all P<0.05). The display clarity of lymph nodes or vascular space of the observation group was 90.00%, which of the control group was 92.50%, there was no statistically significant difference between the two groups (P>0.05). The rates of ground glass shadow, burr of the observation group were 29.32%, 31.58%, respectively, which were significantly lower than those of the control group(41.35%, 47.37%), the differences were statistically significant (χ2=4.21, 6.94, all P<0.05). Other signs detection probability are similar between the two groups and had no statistically significant differences (all P>0.05).@*Conclusion@#The low dose chest CT scan with 16 rows spiral CT has high clinical value, low radiation dose and high diagnostic accuracy in the diagnosis of pulmonary tuberculosis.
RESUMO
Objective To evaluate the clinical value of 16-slice spiral CT low dose chest scanning in the diagnosis of pulmonary tuberculosis.Methods From June 13,2014 to June 20,2017,80 patients with pulmonary tuberculosis in Yuyao People＇s Hospital were selected in the study.The conventional chest dose( control group) and low dose(observation group) of 16 -slice spiral CT were used.The diagnostic accuracy,radiation dose of the two methods,lymph node or vascular space display and image features were observed.Results The diagnostic accuracy of the two methods had no statistically significant difference (P>0.05).The dose product length,the CT dose index in the observation group were (32.98 ± 2.57) mGycm,(44.29 ± 3.47),respectively,which were significantly lower than those in the control group[(127.66 ± 5.03)mGycm,(44.29 ± 3.47)] ( t=106.01,21.05,all P<0.05).The display clarity of lymph nodes or vascular space of the observation group was 90.00% ,which of the control group was 92.50% ,there was no statistically significant difference between the two groups (P>0.05).The rates of ground glass shadow,burr of the observation group were 29.32% ,31.58% ,respectively,which were significantly lower than those of the control group (41.35% ,47.37% ),the differences were statistically significant ( χ2 =4.21,6.94,all P <0.05).Other signs detection probability are similar between the two groups and had no statistically significant differences (all P>0.05).Conclusion The low dose chest CT scan with 16 rows spiral CT has high clinical value, low radiation dose and high diagnostic accuracy in the diagnosis of pulmonary tuberculosis.
RESUMO
OBJECTIVE:To study the inhibitory effects of recombinant adenovirus Ad-GFP-C197, which prepared by adenovirus vector system-loading human telomerase reverse transcriptase(hTERT)C fragment(C197),on the proliferation of 3 kinds of tumor cells in vitro. METHODS:Ad-GFP-C197 was amplified and purified with HEK293 cells. Human gastric cancer cells SGC7901,human breast cancer cells MCF7 and human colorectal cancer cells CaCO2 were infected by Ad-GFP-C197 respectively. Using blank adenovirus carrier (Ad-GFP) as reference,the protein expression of C197 in 3 kinds of tumor cells infected by Ad-GFP-C197 was detected by Western blot assay. The inhibitory effects of Ad-GFP-C197 on 3 kinds of tumor cells were detected by MTT assay. The cell proliferation curve was drawn and the proliferation inhibition rate was calculated. RESULTS:The protein expression of C197 was not detected in 3 kinds of tumor cells infected by Ad-GFP,while significant protein expression of C197 was found in above cells infected by Ad-GFP-C197. The proliferation curves of the 3 kinds of tumor cells infected by Ad-GFP-C197 were significantly inhibited with the time extended,and the proliferation inhibitory rate reached 37.31%-41.42%. CONCLUSIONS:Ad-GFP-C197 shows significant inhibitory effects on the proliferation of SGC7901,MCF7 and CaCO2 cells, which is rapid to make up for the slow effect of other telomerase inhibitors.
RESUMO
OBJECTIVE:To systematically review the efficacy and safety of cinacalcet in the treatment of hemodialysis pa-tients with secondary hyperparathyroidism,and provide evidence-based reference for the clinical treatment. METHODS:Retrieved from Medline,Cochrane Library,EMBase and CBM,randomized controlled trials(RCT)about cinacalcet in the treatment of he-modialysis patients with secondary hyperparathyroidism (SHPT) were collected. Meta-analysis was performed by using Rev Man 5.3.5 software after data extract and quality evaluation by Cochrane systematic Rev Man 5.3.5. RESULTS:Totally 7 RCTs were en-rolled,involving 1 987 patients. Results of Meta-analysis showed cinacalcet can significantly reduce the rate of surgical parathyroid-ectomy[RR=0.23,95%CI(0.06,0.89),P=0.03],incidence of fracture[RR=0.26,95%CI(0.12,0.60),P=0.002] and increase the incidences of hypocalcemia[RR=9.81,95%CI(3.92,4.59),P<0.001],nausea[RR=1.97,95%CI(1.58,2.46),P<0.001] and vomit-ing[RR=1.91,95%CI(1.50,2.42),P<0.001],while it showed no significant effect on the the incidence of all-cause mortality and cardiovascular death. CONCLUSIONS:The clinical efficacy of cinacalcet in the treatment of hemodialysis patients with secondary hyperparathyroidism is good,but there are common adverse reactions such as nausea and vomiting,hypocalcemia.
RESUMO
Objective To compare volumetric‐modulated arc therapy(VMAT) with intensity‐modulated radiation therapy (IMRT) for brain metastases with regard to the dosimetric character .Methods Sixty patients who were diagnosed with brain me‐tastases were included in this study .The target area received two dose levels using late addition amount technique ,WBRT (30 Gy/10 F) with following addition (20 Gy/10 F) to 59 Gy .For a fair comparison ,VMAT and IMRT treatment plans were respectively designed for every patient with the same dosimetric constraints .Dosimetric comparisons between VMAT and IMRT plans were ana‐lyzed to evaluate :target coverage and homogeneity ,conformity of PTV ;sparing of OARs ;monitor units (MUs) .Results Two treatment plans all reached the treatment need .When compared with IMRT ,there was no significant difference in Dmean of eyeball , len ,optic never ,visual chiasma ,parotid ,brain stem ,and external auditory canal of VMAT (P>0 .05) .The Dmax of eyeball ,len ,pa‐rotid ,and external auditory canal of VMAT were lower than that in IMRT group (P<0 .05) .The VMAT group has the less MUs (P=0 .017) and less treatment time .Conclusion VMAT can reach the big‐dose radiotherapy need on brain metastases clinically . There are no significant diffference between VMAT and IMRT on Dmax ,Dmean ,CI ,and HI .The Dmax of eyeball ,len ,parotid ,and external auditory canal of VMAT were lower than that in IMRT group .The VMAT can reduce the radiotherapy time .
RESUMO
Fibroblast growth factor -21 (FGF-21) is a recently discovered metabolic regulation factor, regulating glucose and lipid metabolism and increasing insulin sensitivity. FGF-21 is expected to be a potential anti-diabetic drug. Expression of FGF-21 as inclusion bodies has advantages for high yield and purity, but the bioactivity of the protein is almost totally lost after denature and renature. That is why FGF-21 is currently expressed in soluble form. As a result, the yield is considerably low. In this study, we used SUMO vector to express SUMO-human FGF-21 (SUMO-hFGF-21) in form of inclusion body. We optimized the culture conditions to increase the yield of the bioactive human fibroblast growth factor-21. We applied the hollow fiber membrane filtration column to enrich the bacteria, wash, denature and renature inclusion bodies. After affinity and gel filtration chromatography, we examined the hypoglycemic activity of FGF-21 by the glucose uptake assay in HepG2 cells. We also detected the blood glucose concentration of type 2 diabetic db/db model mice after short or long-term treatment. The results show that the yield of ihFGF-21 was 4 times higher than that of shFGF-21. The yield was 20 mg/L for ihFGF-21 vs. 6 mg/L for shFGF-21. The purity of ihFGF-21 was above 95%, while shFGF-21 was 90%. Compared with the traditional method of extracting inclusion bodies, the production cycle was about three times shortened by application of hollow fiber membrane filtration column technology, but the bioactivity did not significantly differ. This method provides an efficient and cost-effective strategy to the pilot and industrial production of hFGF-21.
Assuntos
Animais , Humanos , Camundongos , Bactérias , Metabolismo , Diabetes Mellitus Experimental , Tratamento Farmacológico , Modelos Animais de Doenças , Fatores de Crescimento de Fibroblastos , Vetores Genéticos , Glucose , Metabolismo , Células Hep G2 , Hipoglicemiantes , Corpos de Inclusão , Metabolismo , Proteínas Recombinantes de Fusão , Proteínas Modificadoras Pequenas Relacionadas à UbiquitinaRESUMO
Previous studies proposed that the synergistic effect of fibroblast growth factor-21 (FGF-21) and insulin may be due to the improvement of insulin sensitivity by FGF-21. However, there is no experimental evidence to support this. This study was designed to elucidate the mechanism of synergistic effect of FGF-21 and insulin in the regulation of glucose metabolism. The synergistic effect of FGF-21 and insulin on regulating glucose metabolism was demonstrated by investigating the glucose absorption rate by insulin resistance HepG2 cell model and the blood glucose chances in type 2 diabetic db/db mice after treatments with different concentrations of FGF-21 or/and insulin; The synergistic metabolism was revealed through detecting GLUT1 and GLUT4 transcription levels in the liver by real-time PCR method. The experimental results showed that FGF-21 and insulin have a synergistic effect on the regulation of glucose metabolism. The results of real-time PCR showed that the effective dose of FGF-21 could up-regulate the transcription level of GLUT1 in a dose-dependent manner, but had no effect on the transcription level of GLUT4. Insulin (4 u) alone could up-regulate the transcription level of GLUT4, yet had no effect on that of GLUT1. Ineffective dose 0.1 mg kg(-1) FGF-21 alone could not change the transcription level of GLUT1 or GLUT4. However, when the ineffective dose 0.1 mg x kg(-1) FGF-21 was used in combination with insulin (4 u) significantly increased the transcription levels of both GLUT1 and GLUT4, the transcription level of GLUT1 was similar to that treated with 5 time concentration of FGF-21 alone; the transcription level of GLUT4 is higher than that treated with insulin (4 u) alone. In summary, in the presence of FGF-21, insulin increases the sensitivity of FGF-21 through enhancing GLUT1 transcription. Vice versa, FGF-21 increases the sensitivity of insulin by stimulating GLUT4 transcription in the presence of insulin. FGF-21 and insulin exert a synergistic effect on glucose metabolism through mutual sensitization.
RESUMO
BACKGROUND:The etiological factor for rheumatoid arthritis remains unclear, but the effects of nuclear factor-κB on the onset of rheumatoid arthritis have been gradual y paid great attention by rheumatologists. OBJECTIVE:By using the RNA interference (RNAi) technique to block the signal pathway of nuclear factor-κB mRNA of human rheumatoid arthritis synovial cells, this study explored its application prospect in the treatment of rheumatoid arthritis. METHODS:The synovial cells were isolated, digested, and cultured for further use. In accordance with the design principle of smal interfering RNA (siRNA), target sequences of siRNA of nuclear factor-κB were identified, and siRNA expression vector of nuclear factor-κB was synthesized and constructed. The four pGenesil-1/nuclear factor-κB siRNA expression vectors were transfected into the first passage of synovial cells that wel grew. Blank and negative control groups were set. cells at 12, 24, 48, 72 hours, 5 and 7 days after transfection were col ected, and RNA was extracted. Intracellular nuclear factor-κB mRNA expression levels were measured, and siRNA plasmid vector that could effectively inhibit nuclear factor-κB mRNA expression was screened out. RESULTS AND CONCLUSION:Nuclear factor-κB highly expressed in synovial cells after human rheumatoid arthritis. 3#pGenesil-1/nuclear factor-κB apparently suppressed nuclear factor-κB mRNA expression in synovial cells with human rheumatoid arthritis. RNAi technique blocked nuclear factor-κB mRNA expression. Therefore, the block of nuclear factor-κB signal pathway might be a good target for rheumatoid arthritis gene therapy.
RESUMO
Objective:To clarify the effect of voltage-gated proton channel 1 (Hv1) on the migration and invasion of breast cancer cells. Methods:The protein expression of Hv1 was detected in human breast cancer cell lines with different metastatic abilities. SiRNA technique was used to down-regulate the expression of Hvl in breast cancer MDA-MB-231 cells. Scratch and matrigel invasion methods were used to observe the effect of Hvl on the migration and invasion of breast cancer cells, and the relevant molecular mechanism was explored. Results:Hv1 was highly expressed in the highly metastatic breast cancer cell line MDA-MB-231. Hvl was more highly expressed in MDA-MB-231 cells with higher metastatic ability. The SiRNA sequence target at Hvl inhibited Hvl expression. Scratch and matrigel invasion experiments showed that the migration and invasion of MDA-MB-231 cells were significantly attenuated when Hv1 was knocked down by siRNA targeting Hv1. Zymography experiment on matrix metalloproteinase indicated that the enzyme activities of MMP-2 markedly decreased. Conclusion:Hv1 promoted the migration and invasion ability of breast cancer cells.
RESUMO
Objective To construct a human phage single chain-antibody library, and to sieve out the antibody ScFv against lung cancer from the library. Methods Total RNA was abstracted from lymph node tissue of the lung cancer, and was used to amplify V_H and V_L gene by RT-PCR. V_H and V_L were joined by a DNA linker by SOE-PCR to form the single chain variable fragment ( ScFv) gene. ScFv gene was coloned into the phage vector pCANT-AB5E. Panning against lung cancer cell line A549 was performed and positive clones were chosen for soluble expression. Results A recombination phage single chain-antibody library was constructed. After 4 rounds panning, the number of eluted phages increased by 115 times. Positive reactions to A549 were detected in 7 of 10 random clones. The human ScFvs against lung cancer were produced and confirmed by SDS-PAGE and ELISA analysis. Conclusion ScFvs against lung cancer were acquired by the construction of phage single chain-antibody library. The soluble ScFvs has specificall avidity to human lung cancer cells.
RESUMO
Objective To construct a human phage single chain-antibody library,and to sieve out the antibody ScFv against lung cancer from the library.Methods Total RNA was abstracted from lymph node tissue of the lung cancer,and was used to amplify VH and VL gene by RT-PCR. VH and VL were joined by a DNA linker by SOE-PCR to form the single chain variable fragment (ScFv) gene. ScFv gene was coloned into the phage vector pCANTAB5E. Panning against lung cancer cell line A549 was performed and positive clones were chosen for soluble expression.Results A recombination phage single chain-antibody library was constructed. After 4 rounds panning,the number of eluted phages increased by 115 times. Positive reactions to A549 were detected in 7 of 10 random clones. The human ScFvs against lung cancer were produced and confirmed by SDS-PAGE and ELISA analysis.Conclusion ScFvs against lung cancer were acquired by the construction of phage single chain-antibody library. The soluble ScFvs has specificall avidity to human lung cancer cells.