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@#[摘 要] 目的:探讨碱性亮氨酸拉链转录因子ATF样蛋白3(BATF3)在肾透明细胞癌(ccRCC)组织中的表达及其调控ccRCC细胞恶性生物学行为的分子机制。方法:收集2019年3月至2022年1月间在中国人民解放军联勤保障部队第九八〇医院手术切除的64例ccRCC组织和相应癌旁组织,qPCR法检测ccRCC组织、癌旁组织和肾癌ACHN、786-O细胞中BATF3 mRNA的表达,免疫组化检测ccRCC组织、癌旁组织中BATF3蛋白的表达,并分析其与临床病理特征的关系。构建BATF3敲减及过表达质粒,分别转染786-O、ACHN细胞,通过MTS法、Transwell实验检测BATF3对786-O或ACHN细胞增殖、迁移、侵袭能力的影响,qPCR法检测敲减或过表达BATF3对786-O或ACHN细胞EMT相关基因表达的影响,CHIP、双荧光素酶报告基因实验检测BATF3是否与波形蛋白(VIM)启动子区结合并调控其转录,MTS法、Transwell实验检测同时过表达BATF3及敲减VIM对786-O细胞增殖、迁移、侵袭能力的影响。结果:与癌旁组织比较,ccRCC组织中BATF3的mRNA和蛋白均呈高表达(均P<0.01),并且BATF3 mRNA与ccRCC的分化程度和TNM分期密切关联(均P<0.01);与正常肾上皮细胞293T相比,BATF3在ACHN及786-O细胞中均呈高表达(均P<0.01)。敲减BATF3表达均能明显抑制786-O细胞的增殖、迁移、侵袭能力(均P<0.01),过表达BATF3则均能促进ACHN细胞的增殖、迁移和侵袭能力(均P<0.01),敲减或过表达BATF3能抑制786-O细胞或促进ACHN细胞的EMT相关基因的表达(均P<0.01)。BATF3可与VIM启动子区的位点结合,直接调控VIM的转录表达。同时过表达BATF3及敲减VIM可逆转过表达BATF3对786-O细胞增殖、迁移、侵袭能力的影响。结论:BATF3在ccRCC组织中呈高表达,并与其分化程度和TNM分期密切关联;BATF3通过调控VIM的表达影响ACHN、786-O细胞的恶性生物学行为,其可作为临床治疗ccRCC的潜在靶点。
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@#Objective To investigate the underlying drug enhancement mechanisms of the Chuanwu (Aconiti Radix) and Huangqi (Astragali Radix) combination and toxicity reduction of Chuanwu combined with Gancao (Glycyrrhizae Radix et Rhizoma) in Wutou Decoction (乌头汤, WTD), and to elucidate the compatibility principle. Methods The active compounds and potential effective targets of the selected combinations were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Traditional Chinese Medicines Integrated Database (TCMID). The toxicity of Chuanwu (Aconiti Radix) was investigated by selecting all five toxic compounds from the literature and the TCMSP database, and obtaining their targets through SwissTargetPrediction. Targets related to rheumatoid arthritis (RA) were searched using DisGeNET, GenCards, and Online Mendelian Inheritance in Man (OMIM). Mutual targets between the drug pairs and RA were selected as potential RA therapy targets. The medicinally active compound-target network was constructed using Cytoscape 3.9.0. Gene ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were performed using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) platform. Results We obtained 191 active compound targets for Gancao (Glycyrrhizae Radix et Rhizoma), 171 for Huangqi (Astragali Radix), and 103 for Chuanwu (Radix Aconiti) (hypoaconitine’s target was obtained through literature and SwissTargetPrediction). A total of 5872 genes were obtained for RA. A drug-active compound-target network involving 13 effect-enhancing and nine toxicity reduction targets was constructed. PGR was the main effect enhancement target, and KCNH2 was the main toxicity reduction target. The effect-enhancing targets were related to 23 GO terms (such as positive regulation of transcription from RNA polymerase II promoter, steroid hormone-mediated signaling pathway, plasma membrane, and protein binding) (P < 0.01), and 13 KEGG pathways related to synergism [such as estrogen signaling pathway, cholinergic synapse, and phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway]. The toxicity reduction targets were related to 28 GO terms (mainly involes G-protein coupled receptor signaling pathway, plasma membrane, and drug binding) (P < 0.01), and five KEGG pathways related to toxicity reduction (cholinergic synapse, calcium signaling pathway, regulation of actin cytoskeleton, neuroactive ligand-receptor interaction, and serotonergic synapse). Conclusion The combination of Chuanwu (Aconiti Radix) and Huangqi (Astragali Radix) plays an important effect-enhancing role in WTD and involves the estrogen and PI3K/Akt signaling pathways, with PGR as the core. The Chuanwu (Aconiti Radix) and Gancao (Glycyrrhizae Radix et Rhizoma) combination decreases toxicity in WTD and is associated with the cholinergic synapse and calcium signaling pathways, with KCNH2 as the core.
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Snakebites remain a major life-threatening event worldwide. It is still difficult to make a positive identification of snake species by clinicians in both Western medicine and Chinese medicine. The main reason for this is a shortage of diagnostic biomarkers and lack of knowledge about pathways of venom-induced toxicity. In traditional Chinese medicine, snakebites are considered to be treated with wind, fire, and wind-fire toxin, but additional studies are required. Methods: Cases of snakebite seen at the Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine were grouped as follows: fire toxin - including four cases of bites by Agkistrodon acutus and three bites by Trimeresurus stejnegeri - and wind-fire toxin - four cases of bites by vipers and three bites by cobras. Serum protein quantification was performed using LC-MS/MS. Differential abundance proteins (DAPs) were identified from comparison of snakebites of each snake species and healthy controls. The protein interaction network was constructed using STITCH database. Results: Principal component analysis and hierarchical clustering of 474 unique proteins exhibited protein expression profiles of wind-fire toxins that are distinct from that of fire toxins. Ninety-three DAPs were identified in each snakebite subgroup as compared with healthy control, of which 38 proteins were found to have significantly different expression levels and 55 proteins displayed no expression in one subgroup, by subgroup comparison. GO analysis revealed that the DAPs participated in bicarbonate/oxygen transport and hydrogen peroxide catabolic process, and affected carbon-oxygen lyase activity and heme binding. Thirty DAPs directly or indirectly acted on hydrogen peroxide in the interaction network of proteins and drug compounds. The network was clustered into four groups: lipid metabolism and transport; IGF-mediated growth; oxygen transport; and innate immunity. Conclusions: Our results show that the pathways of snake venom-induced toxicity may form a protein network of antioxidant defense by regulating oxidative stress through interaction with hydrogen peroxide.(AU)
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Animales , Venenos de Serpiente , Biomarcadores , Estrés Oxidativo , Peróxido de Hidrógeno , Antioxidantes , Trimeresurus , Proteoma/análisisRESUMEN
@# Objective: To explore the relationship between monocyte-to-lymphocyte ratio (MLR) in peripheral blood of patients with pulmonary sarcomatoid carcinoma (PSC) and their clinicopathological features and prognosis, and to investigate its clinical significance. Methods: A retrospective analysis was carried out to analyze the complete case data of 80 patients with PSC from October 2010 to April 2017 in Tianjin Cancer Hospital (monocyte and lymphocyte counts of peripheral blood, clinicopathological features, and survival follow-up). The receiver operating curve (ROC) was used to determine the best cut-off value of MLR for the prediction of overall survival time (OS). The patients were divided into high MLR group and low MLR group. Kaplan-Meier method was used to calculate OS and draw survival curves. The Log-Rank test was used to compare the difference in OS between the two groups. The variables with statistical significance in univariate analysis were included into the COX risk regression model to verify and calculate thehazard ratio (HR)and 95% confidence interval (95%CI). Results: The absolute median values of monocytes and lymphocytes were 0.63×109/L and 1.84×109/L, respectively. The best cut-off value of MLR is 0.44. Univariate analysis shows that MLR≥0.44 (P<0.01), no radical surgery (P<0.01), clinical stage Ⅲ+Ⅳ (P<0.01), tumor maximal diameter > 3 cm (P<0.01), and LDH>247 U /L (P<0.01) are the poor prognostic factors affecting overall survival. Multivariate analysis shows that MLR≥0.44(HR=3.554; 95%CI=1.671-6.125; P<0.01), and clinical stage Ⅲ+Ⅳ(HR=3.275; 95%CI=2.047-9.399; P<0.01) are the independent risk factors for the overall survival of PSC, and radical surgery is an independent protective factor affecting the overall survival of PSC(HR=0.360; 95%CI=0.195-0.848; P<0.01). Conclusion: High MLR is an independent risk factor for poor prognosis in patients with PSC.
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OBJECTIVE To explore the estrogen- like neuroprotective effects and the related mechanism of quercetin by using PC12 cells induced with Aβ25-35, provided thought and strategy for the drug therapy of AD. METHODS Cells were cultured with Aβ25- 35 for 24 h, 17β-estradiol (0.1 μmol·L- 1), genistein(50 μmol·L-1) and three different concentrations of quercetin (200 μmol·L-1, 300 μmol·L-1 and 400 μmol·L-1) were respectively added after 24h. The effects of quercetin on activity of AD model were tested by MTT. Immunohistochemical stain and Western blot were used to detect the expression of estrogen receptors alpha and beta, p-ERK1/2 and apoptosis related proteins.The mechanism of quercetin estrogen-like neuroprotective effects was detected using estrogen receptor antagonist ICI182,780 and MAPKK inhibitor U0126. RESULTS The results revealed thatthe toxic effects showed in a dose-dependent increase of Aβ25- 35 on PC12 cells.Comparing with the control group,cells injury was observed after cultured with 10 μmol·L-1 Aβ25-35 for 24 h(P<0.01). The MTT results showed that 17β-estradiol, genistein and three different concentrations of quercetin could significantly enhance the cell survival rate compared with the model group (P<0.05). Compared with model group,Immunofluorescence and Western blot results show that quercetin could improve the estrogen receptor alpha and p- ERK1/2 protein expression (P<0.05), and the expression of estrogen receptor beta protein is increased without significant difference. And in the Western blot experiments, the ratio of Bcl- 2 and Bax was increased and the expression of Caspase 3 was decreased( P<0.05).When estrogen receptor inhibition ICI182,780 were reduced,the expression of p- ERK1/2 was decreased (P<0.05) and the ratio of Bcl- 2 and Bax was decreased, Caspase 3 protein expression was increased (P<0.05). In addition,pretreatment of cells with U0126 would reduce Bcl-2/Bax ratio and increase Caspase 3 protein expression increased (P<0.05). CONCLUSION Quercetin protected PC12 cells, which suffered from Aβ25- 35-induced cytotoxicity and exert neuroprotective effects. The estrogen-like neuroprotective effect can reduce the apoptosis in the classic estrogen receptor pathway and MAPK pathway. And quercetin can also active MAPK signaling pathways by the mediation of estrogen receptor alpha.
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OBJECTIVE To investigate the effect of quercetin on primary cultured newborn rat cortex neuron cell which is estrogen depletion, and discuss the possible mechanism, to provide new ideas and strategies for developing a drug of neurodegenerative disease. METHODS Rat cortex neurons were isolated from one day old Sprague Dawley rats and treated with estrogen, quercetin and estrogen receptor antagonists (ICI182,780). Cell viability was determined by MTT assay, neurite outgrowth was measured by fluorescent microsope and estrogen receptors were determine by Western blot. RESULTS Quercetin functions like estrogen to increase cortex neuronal cell viability, the Que (50, 100 μmol·L-1) group compared with the control group could significantly improve the activity of the cortical neurons(P<0.05). It can also increase neurite out growth, the Que (50,100 μmol·L-1) group significantly promoted the formation of synapse, most of the neurons were full, and the synapses of neurons became thick, growth, and connect to a dense neural network. And in the Western blot experiments, Que (50, 100 μmol·L-1) group could obviously increase the expression of estrogen receptor alpha protein, in addition, the neural protective effect of quercetin can be inhibited by ICI182,780. CONCLUSION Quercetin like estrogen can protected cortex neuronal and the effect of quercetin on cortex neuronal cells was mediated by estrogen receptor alpha.
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<p><b>OBJECTIVE</b>To investigate the prevalence and risk factors of peripheral arterial disease (PAD) in male Chinese octogenarians and nonagenarians with hypertension.</p><p><b>METHODS</b>Ankle-brachial index (ABI) was measured in the noninvasive vascular laboratory for hypertensive male octogenarians and nonagenarians enrolled from outpatient and inpatient departments. The baseline conditions were investigated using standard questionnaire and by routine physical examinations. PAD was diagnosed when an ABI≤0.9 in either lower extremity.</p><p><b>RESULTS</b>Totally 290 male Chinese octogenarians and nonagenarians [age: (84.61±4.20) years] with hypertension who were receiving antihypertensive therapy were enrolled in this study, among whom 9 men with missing data except age and ABI measurements. The ABI was 0.948±0.258, with the range of highest frequency of 0.91-1.30, and 106 patients were diagnosed as PAD, 182 as non-PAD, and 2 had ABI>1.3. ABI in hypertensive men with PAD were significantly lower than in those without PAD (P<0.05). On the contrary, age, blood urea nitrogen, white blood cell counts, platelets and aspartic transaminase were significantly higher in PAD patients than in non-PAD patients (all P<0.05). The prevalence of PAD in this study population were 36.5%; more specifically, it significantly differed between different subgroups when stratified by age (28.6% vs. 46.3%, below and above 85 years), with and without hypertension (27.5% vs. 40.1%), stroke (44.7% vs. 27.5%), dyslipidemia (41.4% vs. 33.3%), coronary artery disease (44.1% vs. 13.9%), and diabetes mellitus (53.7% vs. 21.8%) (all P<0.05). The prevalences of PAD in hypertensive patients treated with diuretics, calcium antagonists, beta-blocker, or angiotensin receptor antagonist were 41.4%, 36.1%, 22.4%, and 26.8%, respectively. No association was observed between the prevalence of PAD and smoking/alcohol drinking in these subjects. Multivariate analysis showed that age (OR 1.12, 95%CI 1.014-1.238), blood urea nitrogen (OR 1.15, 95%CI 1.025-1.301), aspartic transaminase (OR 1.05, 95%CI 1.005-1.089), diabetes mellitus (OR 4.02, 95%CI 1.797-9.009), coronary artery disease (OR 6.34, 95%CI 1.734-23.214) were strong risk factors of PAD.</p><p><b>CONCLUSION</b>PAD is highly prevalent among aged Chinese hypertensive men, in which age, blood urea nitrogen, aspartic transaminase, diabetes mellitus, coronary artery disease may be involved in the development of this condition.</p>
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Anciano de 80 o más Años , Humanos , Masculino , Pueblo Asiatico , Hipertensión , Enfermedad Arterial Periférica , Epidemiología , Prevalencia , Factores de RiesgoRESUMEN
This study investigated the role of reactive oxygen species (ROS) in the pathogenesis of triptolide-induced renal injury in vivo.Rats were randomly divided into 4 groups (n=5 in each):triptolide group in which the rats were intraperitoneally injected with triptolide solution at a dose of 1 mg/kg of body weight on day 8; control group in which the rats received a single intraperitoneal injection of 0.9% physiological saline on day 8; vitamin C group in which the rats were pretreated with vitamin C by gavage at a dose of 250 mg/kg of body weight per day for 7 days before the same treatment as the control group on day 8; triptolide+vitamin C group in which the rats were first subjected to an oral administration of vitamin C at a dose of 250 mg/kg of body weight per day for 7 days,and then to the same treatment as the triptolide group on day 8.All the rats were sacrificed on day 10.Blood samples were collected for detection of plasma creatinine (Pcr) and plasma urea nitrogen (PUN) concentrations.Both kidneys were removed.The histological changes were measured by haematoxylin-eosin (HE)staining.The production of ROS was determined by detecting the fluorescent intensity of the oxidation-sensitive probe rhodamine 123 in renal tissue.Renal malondialdehyde (MDA) content was measured to evaluate lipid peroxidation level in renal tissue.TUNEL staining was performed to assess apoptosis of renal tubular cells.Renal expression of apoptosis-related proteins Bcl-2,Bax,Bid,Bad,Fas and FasL,as well as corresponding encoding genes were assessed by Western Blotting and real-time PCR.The results showed that triptolide treatment promoted the generation of a great amount of ROS,up-regulated the expression of Bax,Bid,Bad,Fas and FasL at both protein and mRNA levels,as well as the ratio of Bax to Bcl-2,and caused the apoptosis of renal tubular cells and renal injury.However,pretreatment with an antioxidant,vitamin C,significantly reduced the generation of ROS and effectively inhibited the triptolide-induced apoptosis of renal tubular cells and renal injury.It was concluded that ROS plays a critical role in triptolide-induced apoptosis of renal tubular cells and renal injury.The protective administration of vitamin C may help alleviate triptolide-induced renal injury and nephrotoxicity.
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Corneal opacity is one of the most commonly used parameters for estimating postmortem interval(PMI).This paper proposes a new method to study the relationship between changes of corneal opacity and PMI by processing and analyzing cornea images.Corneal regions were extracted from images of rabbits' eyes and described by color-based and texture-based features,which could represent the changes of cornea at different PMI.A KNN classifier was used to reveal the association of image features and PMI.The result of the classification showed that the new method was reliable and effective.
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The aim of the present study was to assess whether Fourier transform infrared spectrometry (FTIR)micro-spectroscopy could produce distinct spectral information on protein of old myocardial infarction(OMI)and to set them as molecular markers to diagnose atypical OMI.Paraffin-embedded heart samples were derived from victims dying of OMI.In combination with histological stain,FTIR and infrared micro-spectroscopy,the characteristics of OMI were analyzed morphologicallyand molecularly.The most relevant bands identified were the amide A,B,Ⅰ and Ⅱ,showing crucial spectral differences between apparent normal region and OMI region,including the peak position blue shift and the increased intensity of OMI,moreover relative increase in a-helix and decrease in β-sheet of protein secondary structures in OMI.Comparing to single spectral band,the I1650/I1550 ratio was increased and rationally used as a molecular marker for diagnosing OMI.These novel preliminary findings supported further exploration of FTIR molecular profiling in clinical or forensic study,and were in accordance with histopathology.
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To study the relationship between changes of microbial ATP in four kinds of murine tissues and the postmortem interval (PMI),healthy SD rats were sacrificed and their muscles,livers,spleens and kidneys were sampled at different postmortem intervals. The concentration of microbial ATP was detected using bioluminescent assay and the data was statistically analyzed. The concentration of microbial ATP in muscle increased with PMI time. The peak appeared at the 7th day after death,and at the 10th day,microbial ATP in muscle tissue increased again. In internal organs,the peaks of microbial ATP were observed at the 8th day after death and the level decreased during 8-10 d. The differences in microbial ATP concentration in liver,spleen and kidney were not statisticallysignificant. During day 0 to day 9 after death,the correlation was best between PMI and microbial ATP in muscle. With PMI as the independent variable,the cubic polynomial regression equation was Y=0.02X3-0.166X2-0.666X+13.412 (R2=0.989,P<0.01). In internal organs,the best correlation was found between PMI and microbial ATP during day 0 to day 10. With PMI as the independent variable,the cubic polynomial regression equation was Y=0.016X3-0.127X2-0.809X+13.324 (R2=0.986,P<0.01). There existed high correlations between PMI and microbial ATP concentration in rat tissues.Since only a small amount of tissue was needed for the detection and the sample was not affected by self-decomposition,the method may extend the time range of PMI estimation.
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This study explored the possibility of using event-related potentials (ERP) for the measurement of picture-recognition memory and examined its correlation with the Chinese Wechsler Memory Scale-revised (WMS-RC) in patients with memory disorder caused by severe traumatic brain injury (sTBI). The subjects included 20 sTBI patients with memory disorder and 22 healthy individuals. Memory function was measured by using WMS-RC. Behavioral and ERP responses were recorded on-line during performance on a battery of picture recognition and the responses were analyzed off-line for recognition memory effects. Mean memory quotient (MQ) of patients with sTBI was significantly lower than that of the control group. Mean reaction time (RT) was significantly longer and the mean correctness rate (CR) of picture recognition was significantly lower in sTBI group than that of the controls. In controls, the main components of average ERP of picture recognition includes two positive-going waves, designated as P170 and P500, that appear 170 ms and 500 ms after stimulation when the subject could later successfully recall and recognize the pictures. P500 amplitude of target stimulus was significantly higher than that of non-target stimulus. Compared to controls, P500 responses of sTBI group were significantly delayed in latency (P<0.001) and lower in amplitude (P<0.001). P500 latency showed significant negative correlation with MQ and the scores of "addition", "visual recognition", "picture recall", "visual reproduction" and "tactile memory" in WMS-RC. ERP of picture recognition provides a neurophysiological approach to directly assess memory impairment, and P500 may serve as a helpful index for memory disorder caused by sTBI in forensic practice.
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To study the relationship between the late postmortem interval (PMI) and trimethylamine-nitrogen (TMA-N) in postmortem tissues of cadaver, TMA-N in muscles, livers and kidneys of rats was measured at different postmortem intervals (PMD by using a modified spectrophotometric method. The results indicated that the detection sensitivity of TMA-N was 1 mg/L, and there was a good linear correlation between the value of absorbance (A value) and TMA-N at the concentration of 1-10 mg/L (R2 =0.9991). Although TMA variation in muscles was different from that in inner organs during the time since death, TMA-N changes in cadaver tissues was positively correlated with PMI. During 2 to 7 d since death, the best correlation between PMI and TMA-N concentration was found in muscles.With PMI as an independent variable, the cubic polynomial regression equation was y= --0.457x3+6.519x2-24.574x+27.207 (R2=0.969). During 3 to 8 days since death, PMI was best correlated with TMA-N concentration in inner organs. With PMI as the independent variable, the cubic polynomial regression equation was y=0.509x3-9.153x2+55.727x-95.819 (R2=0.953). It was concluded that TMA-N in tissues could be used as a new estimator for late PMI. The method used in this study offered advantages such as accuracy, sensitivity, little samples required and wide PMI estimation.
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Summary: In order to investigate the effects of aconitine on [Ca2+] oscillation patterns in cultured myocytes of neonatal rats, fluorescent Ca2+ indicator Fluo-4 NW and laser scanning confocal microscope (LSCM) were used to detect the real-time changes of [Ca2+] oscillation patterns in the cultured myocytes before and after aconitine (1.0 μmol/L) incubation or antiarrhythmic peptide (AAP) and aconitine co-incubation. The results showed under control conditions, [Ca2+] oscillations were irregular but relatively stable, occasionally accompanied by small calcium sparks. After incubation of the cultures with aconitine, high frequency [Ca2+] oscillations emerged in both nuclear and cytoplasmic regions, whereas typical calcium sparks disappeared and the average [Ca2+] in the cytoplasm of the cardiomyocyte did not change significantly. In AAP-treated cultures, intraecllular [Ca2+] oscillation also changed, with periodic frequency, increased amplitudes and prolonged duration of calcium sparks. These patterns were not altered significantly by subsequent aconitine incubation. The basal value of [Ca2+] in nuclear region was higher than that in the cytoplasmic region, in the presence or absence of drugs, the [Ca2+] oscillated synchronously in both the nuclear and cytoplasmic regions of the same cardiomyocyte. It was concluded that although oscillating strenuously at high frequency, the average [Ca2+] in the cytoplasm of cardiomyocyte did not change significantly after aconitine incubation, compared to the controls. The observations indicate that aconitine induces the changes in [Ca2+] oscillation frequency other than the Ca2+ overload.
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<p><b>OBJECTIVE</b>To probe into therapeutic effects of Ziwuliuzhu combined selection of the source point and the collateral point on athletic injuries and the state of channels.</p><p><b>METHODS</b>Ninety cases meeting diagnostic criteria were randomly divided into a group of Ziwuliuzhu combined selection of the source point and the collateral point (group A), a group of routine selection of acupoint (group B) and a group of external application of medicine (group C), 30 cases in each group. The electric conduction amount on source points of 12 channels were determined before and after treatment with a point diagnosis and treatment instrument. Changes of the state of channels and clinical therapeutic effects in the 3 groups were investigated after treatment.</p><p><b>RESULTS</b>The therapeutic effect in the group A was better than those in the other two groups (P < 0.05).</p><p><b>CONCLUSION</b>Ziwuliuzhu combined selection of the source point and the collateral point has good effect on pain and other clinical symptoms of athletic injuries, and makes channels of imbalance tend to balance or recover balance.</p>
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Humanos , Puntos de Acupuntura , Terapia por Acupuntura , Traumatismos en Atletas , Medicina , DolorRESUMEN
<p><b>OBJECTIVE</b>To detect the expression and variation of p53 gene during tree shrews' hepatocarcinogenesis induced by hepatitis B virus (HBV) and aflatoxin B1 (AFB1).</p><p><b>METHODS</b>Tree shrews were divided into four groups: the tree shrews were infected with HBV and fed with AFB1 in group A, only infected with HBV in group B, fed with AFB1 alone in group C, and normal control in group D. All the tree shrews were performed liver biopsy every 15 weeks. The tissues of liver and tumor were detected by immunohistochemistry and molecular biotechnologies.</p><p><b>RESULTS</b>(1) The incidence of hepatocellular carcinoma (HCC) in group A (66.7%) was higher than that in Group B and C (30%). HCC appearance in group A was earlier than that in group C (120.0 weeks +/-16.6 weeks vs 153.3 weeks +/-5.8 weeks, t = 3.336, P<0.01). (2) Mutated p53 protein was not found before the 75th week of the experiment in each group. (3) At the 105th week, the expression rates of mutated p53 protein were 78.6%, 60% and 71.4% in group A, B and C respectively, which were much higher than that (10%) in group D (x2 > or = 5.03, P<0.05). An abnormal band of p53 gene was detected in both group A and C. (4) The mutation points of p53 gene in liver cancer of tree shrew were at codon 275, 78 and 13. The nucleotide sequence and amino acids sequence of tree shrew's wild-type p53 showed 91.7% and 93.4% homology with those of human p53 respectively.</p><p><b>CONCLUSIONS</b>There is a remarkable synergistic effect between HBV and AFB1 on HCC. Mutated p53 protein is expressed before HCC occurrence, which promotes the development and progress of HCC. HBV and AFB1 may synergistically induce p53 gene mutation.</p>
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Animales , Aflatoxina B1 , Toxicidad , Carcinoma Hepatocelular , Genética , Cocarcinogénesis , Regulación Neoplásica de la Expresión Génica , Variación Genética , Hepatitis B , Virología , Virus de la Hepatitis B , Neoplasias Hepáticas Experimentales , Genética , Mutación Puntual , ARN Neoplásico , Proteína p53 Supresora de Tumor , Genética , TupaiidaeRESUMEN
<p><b>OBJECTIVE</b>(1) To investigate the expression and gene diversity of the 7 major cancer/testis (CT) antigens, MAGE-1, MAGE-3, MAGE-4, MAGE-10, NY-ESO-1, SSX-2 and SCP-1, in hepatocellular carcinoma (HCC). (2) To analyze the correlations between the clinical characters and CT antigens' expression.</p><p><b>METHODS</b>The cancer and para-cancer tissues were collected from 30 HCC patients. The mRNAs of seven CT antigens were detected by reverse transcription-polymerase chain reaction (RT-PCR) with the specific primers. The PCR products were sequenced to analyze the CT genes.</p><p><b>RESULTS</b>The MAGE-1, MAGE-3, MAGE-4, MAGE-10, NY-ESO-1, SSX-2 and SCP-1 were expressed in 66.7%, 70.0%, 20.0%, 36.7%, 40.0%, 33.3% and 33.3% of the tumor tissues from HCC patients respectively, however, they were not expressed in the para-cancer tissues. Among the 30 patients investigated, 90.0% expressed one CT gene at least, 70.0% expressed two CT genes, and 53.3% expressed three CT genes of the seven CT genes. The coding genes of these CT antigens were highly conserved between in Chinese patients and patients abroad. There were discernible correlations between alpha-fetoprotein level and MAGE-10 or SCP-1 expression level, as well as between average age and MAGE-3 or SSX-2 expression levels (P<0.05).</p><p><b>CONCLUSIONS</b>With a highly conserved coding gene, seven CT antigens were expressed in 20.0% - 70.0% of Chinese HCC patients. CT antigens' expression had correlations with some clinical characters.</p>
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Femenino , Humanos , Masculino , Antígenos de Neoplasias , Genética , Carcinoma Hepatocelular , Genética , Alergia e Inmunología , Regulación Neoplásica de la Expresión Génica , Genética , Neoplasias Hepáticas , Genética , Alergia e Inmunología , Antígenos Específicos del Melanoma , Proteínas de Neoplasias , Genética , ARN Mensajero , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To understand the molecular mechanism and find out the responsible genes for liver cancer by exploring the regulation of gene expression during hepatocarcinogenesis in tree shrew induced by aflatoxin B1 (AFB1).</p><p><b>METHODS</b>The tissues from tree shrew of different stages during the pathogenesis and development of hepatocellular carcinoma (HCC), liver cancer tissue, para-cancerous tissues, pre-cancerous liver tissues, liver tissues of the same stage from normal controls and the liver tissues taken before AFB1-treatment were analyzed for gene expression by cDNA array.</p><p><b>RESULTS</b>Four patterns of gene expression were observed during AFB1-induced hepatocarcinogenesis. They were: genes up-regulated in HCC tissue and para-cancerous tissue, especially in HCC tissues; genes with similar expressing level in both HCC tissue and para-cancerous tissue, but higher than that in pre-cancerous tissue; genes down-regulated in HCC tissue; genes up-regulated before HCC appeared but down-regulated after HCC appeared.</p><p><b>CONCLUSION</b>Dynamic observation of gene expression will be beneficial to elucidate the mechanisms of AFB1- induced hepatocarcinogenesis and locate the responsible genes.</p>
Asunto(s)
Animales , Aflatoxina B1 , Toxicidad , Perfilación de la Expresión Génica , Neoplasias Hepáticas Experimentales , Genética , Análisis de Secuencia por Matrices de Oligonucleótidos , Métodos , TupaiidaeRESUMEN
<p><b>OBJECTIVE</b>To examine sensitivity of the tree shrews and Macaca assamensis to human hepatitis B virus (HHBV) by serologic methods.</p><p><b>METHODS</b>Totally 233 tree shrews and 28 Macaca assamensis were inoculated with human sera containing HBV. After inoculation, the sera were collected weekly from them and HBV markers were detected with HBV ditecting ELISA kits.</p><p><b>RESULTS</b>Ninety percent of the tree shrews developed acute infection, among them, 44.4 % persisted for over one year, 33.3% of them developed chronic infection persisted for 2 years and one month; the persistence of HBV in Macaca assamensis was much shorter.</p><p><b>CONCLUSION</b>These data clearly indicated that tree shrew may be used as an animal model for study of chronic HBV infection, whereas, Macaca assamensis, showed only a transient sensitivity to HHBV.</p>
Asunto(s)
Animales , Femenino , Humanos , Masculino , Modelos Animales de Enfermedad , Hepatitis B , Sangre , Alergia e Inmunología , Virología , Antígenos de Superficie de la Hepatitis B , Sangre , Antígenos e de la Hepatitis B , Sangre , Virus de la Hepatitis B , Macaca , TupaiidaeRESUMEN
<p><b>BACKGROUND</b>To serially observe the pathologic changes in livers of tree shrews and macaca assamensises infected with HHBV.</p><p><b>METHODS</b>10 adult tree shrews and 28 macaca assamensises were inoculated with HBV rich human sera. The liver of the animals were regularly biopsied. The liver samples were examined histopathologically by HE staining. Some samples were stained for HBsAg by immunohistochemistry (IH), and HBV DNA by in situ hybridization (ISH).</p><p><b>RESULTS</b>HBsAg in 80% of tree shrews infected with HHBV can be detected by IH, HBV DNA in 50% of those can be found by ISH.The positive rates of HBsAg in macaca assamensises' livers were 25% by IH, none HBV DNA was detected.</p><p><b>CONCLUSION</b>The tree shrew model seems to be applicable for the research of human hepatitis B.</p>