ABSTRACT
The early diagnosis and effective treatment of hepatocellular carcinoma (HCC) still remains a difficult problem that plagues the medical community. Exosomes are microvesicles with a diameter of 40~100 nm, and contains proteins, lipids and nucleic acids (mRNAs, lncRNAs, circRNAs, and microRNAs). They serve as an information exchange carrier, and play an important role in regulating and controlling the biomolecular function to maintain the stability of the intracellular environment. The function of exosomes in HCC includes intercellular communication, neoangiogenesis, cancer cell metastasis and multidrug resistance, which mediates the transformation of microRNAs (miRNA) and regulate the microenvironment of tumor progression, and then affect the pathophysiological behavior of cancer cells. Exosome-derived miRNA can be used for HCC monitoring or potential specific markers of early diagnosis. In addition, with the development and application prospects it could be a therapeutic goal for HCC. This paper summarizes the recent progress in the study of HCC-derived exosomal miRNA.
ABSTRACT
Objective@#To investigate the Wnt3a expression in tissues of HCC and its gene knockout on effects of HepG2 cell proliferation or xenograft tumor growth.@*Methods@#Hepatic Wnt3a expressions in 87 HCC and their matched surrounding tissues were observed by tissue microarray and immunohistochemistry for analyzing its clinicopathological characteristics; Wnt3a-knockout HepG2 cell lines were established by Crispr/cas9-sgRNA system and genomic cleavage efficiency was verified at gene level by surveyor assay. The relative proteins were confirmed by Western blotting; Cell Counting Kit-8 assay was used to examine cell proliferation after knocking-out Wnt3a successfully, and the nude mice HepG2 cell xenograft tumors delete that the relationship between Wnt3a and HCC growth.@*Results@#The positive Wnt3a with brown staining particles was mainly distributed in cytosol and membrane of hepatocytes. The incidence of hepatic Wnt3a expression in cancerous tissues (95.4%) was significantly higher (χ 2 = 47.754, P < 0.001) than that in their surrounding tissues (49.4%). The high Wnt3a expression was 70.1% in the HCC and only 14.9% in the surrounding tissues. High Wnt3a expression was associated with poorly-differentiated grade, liver cirrhosis, HBV infection, portal vein invasion, TNM stage and 5-year survival rate. After knocked-out by Crispr/cas9-sgRNA system successfully, Wnt3a expression was down-regulated significantly at gene or protein level. Key molecule β-catenin in cytoplasma was obviously inhibited. HepG2 cell lines proliferation was suppressed in time-dependent manner. The nude mice HepG2 cell xenograft tumors confirmed that the knock-out of Wnt3a could significantly supressed HCC growth with slower speed (t = 6.418, P < 0.001), smaller volume(869.4 ± 222.5 mm3 vs 355.0 ± 99.9 mm3, t = 5.168, P < 0.001), and lighter weight (0.88 ± 0.20 g vs 0.35 ± 0.11 g, t = 5.628, P < 0.001)compared with the control group.@*Conclusion@#Abnormal expression of Wnt3a could be expected as a promising target for HCC gene therapy.
ABSTRACT
Objective@#To quantitatively detect CD44 expression in patients with nonalcoholic fatty liver disease (NAFLD) for comparative analysis.@*Methods@#Patients with chronic liver diseases accompanied with or without NAFLD, including chronic hepatitis B, cirrhosis and hepatocellular carcinoma after chronic hepatitis B, and healthy blood donors as normal controls who admitted to the Affiliated Hospital of Nantong University from May to October 2018 were selected. The proportion of CD44 positive cells was analyzed by flow cytometry. CD44 level was quantified by an enzyme-linked immunosorbent assay, and the biochemical indicators such as serum aspartate aminotransferase, alanine aminotransferase activity, total cholesterol and triglyceride were routinely analyzed. The cancerous and adjacent cancerous tissues of patients accompanied with or without NAFLD were collected by self-matching method and analyzed by immunoblotting and histochemistry and compared by CD44 integrated optical density. Image-Pro Plus version 6.0, Image J, GraphPad Prism 5.0, Photoshop, Microsoft Excel and IBM SPSS statistics 23 were used to analyze and draw pictures. An independent sample t-test was used to compare the differences between groups.@*Results@#Patients accompanied with NAFLD had hepatocyte injury and dyslipidemia. NAFLD and chronic liver disease patients had significantly elevated serum CD44 levels than normal control group (P < 0.01). CD44 positive lymphocyte ratio was 78.19 % ± 16.33 % in NAFLD patients and 68.47% ± 20.91% in chronic hepatitis B group, which was higher than the control group (46.51% ± 20.52%). Chronic hepatitis B group with steatosis had significantly higher CD44 concentration (181.42 ± 49.36) ng/ml than chronic hepatitis B group (142.52 ± 53.87) ng/ml and normal control group (99.47 ± 15.23) ng/ml. CD44/GAPDH ratio in the liver cancer group (1.306 ± 0.614) was significantly higher than paracancerous group (0.477 ± 0.291) and the difference between the two groups was statistically significant (t = 3.451, P = 0.004). The integrated optical density of CD44 in the NAFLD-related liver cancer and paracancerous group were 25.721 ± 5.881 and 14.155 ± 4.001 and the difference between the groups was statistically significant (t = 14.544, P < 0.001). The pathological features of high expression of CD44 in patients with hepatocellular carcinoma were significantly correlated with HBV infection, tumor size, single/multi-center, and lymph node metastasis, degree of differentiation, TNM grade, Child-Pugh score, portal vein tumor thrombus and extrahepatic metastasis. HCC patients with NAFLD had significantly higher serum CD44 (234.62 ± 69.40) ng/ml than patients without NAFLD (186.49 ± 58.89) ng/ml (t = -3.191, P = 0.002), but there was no statistically significant difference in the clinicopathological characteristics between the high/low CD44 groups of HCC patients with NAFLD.@*Conclusion@#The results suggest that CD44 is abnormally activated and its mechanism may play an important role in the progression of NAFLD.
ABSTRACT
Objective To investigate the safety, effectiveness, and outcome influencing factors of alteplase intravenous thrombolytic therapy in patients with wake-up ischemic stroke (WUIS) under the guidance of MRI. Methods From May 2016 to June 2018, patients with WUIS admitted to the Department of Neurology and Emergency Center of Dongping County People's Hospital were enrolled retrospectively. Patients with WUIS were examined with emergency craniocerebral MRI. Patients with diffusion-weighted imaging and fluid attenuated inversion recovery sequence (DWI/FLAIR) mismatch were treated with alteplase intravenous thrombolysis, and the DWI/FLAIR match patients were not treated with it. The changes of the NIHSS scores at 24 h after admission compared with baseline and the modified Rankin Scale (mRS) score at 90 d were used to analyze the efficacy of treatment. The incidence of complications of intracerebral hemorrhage within 24 h after admission was used to analyze the safety of treatment. Results A total of 65 patients with WUIS were enrolled. Among them, DWI/FLAIR of 30 patients did not match, and DWI/ FLAIR of 35 matched. There was no significant difference in demographic and baseline clinical data between the 2 groups. Compared with the baseline, the NIHSS score was significantly improved at 24 h after admission in the DWI/FLAIR mismatch group (P < 0. 05), whereas the DWI/FLAIR match group was not. The NIHSS score of the DWI/FLAIR mismatch group at 24 h was significantly lower than that of the DWI/FLAIR match group (6. 35 ±2. 56 vs. 9. 69 ±3. 04; t = 4. 702, P = 0. 001), and the good outcome rate at 90 d (mRS score 0-2) was significantly higher than the DWI/FLAIR match group (66. 7％ vs. 22. 7％; χ2 = 5. 634, P = 0. 018 ). Multivariate logistic regression analysis showed that alteplase intravenous thrombolysis (odds ratio [OR] 1. 26, 95％ confidence interval [CI] 1.11-1. 43; P < 0. 001) was a good independent protective factor of the outcome, but hypertension (OR 0. 87, 95％ CI 0. 81-0. 92; P < 0. 001), diabetes (OR 0. 61, 95％ CI 0. 52-0. 72; P < 0. 001), and large-artery atherosclerosis (OR 0. 74, 95％ CI 0. 67-0. 82; P < 0. 001) were the independent risk factors for poor outcome. Two patients had intracerebral hemorrhage within 24 h after admission in the DWI/FLAIR mismatch group and the DWI/FLAIR match group did not have any intracerebral hemorrhage, but there was no significant difference between the 2 groups. Conclusion Intravenous thrombolysis with alteplase was safe and effective for DWI/FLAIR mismatch patients with WUIS.
ABSTRACT
Objective To investigate the risk factors and anticoagulation parameters in patients of acute exacerbation of chronic obstructive pulmonary disease (AECOPD) combined with deep venous thrombosis (DVT). Methods All of 110 AECOPD patients were divided into two groups according to Doppler examination of lower extremities: DVT group and non-DVT group. The risk factors and anticoagulation parameters were compared. Results Twelve cases (10.9%) were in DVT group,of whom 2 cases (1.8%)had pulmonary embolism. The rate of lying in bed > 3 d, smoke, mechanical ventilation, hospital stays and the levels of PaCO2 were significantly higher in DVT group than those in non-DVT group (P < 0.01 or <0.05 ). In DVT group, the activity of antithrombin Ⅲ and the level of protein S decreased (P < 0.05 ), and the level of D-Dimer increased (P < 0.05). Conclusions Long-term bed, smoke and mechanical ventilation requirement are the main risk factors of DVT in patients with AECOPD. Respiratory failure (type Ⅱ ) is easier to combine with DVT. Abnormality of coagulation and fibrinolytic system exists in AECOPD with DVT.
ABSTRACT
BACKGROUND: Status epilepticus can result in neuronal injury.OBJECTIVE: To observe the mitochondrial ultrastructural damage and the changes of Fas, Bax and Caspase-3 expressions in hippocampal CA3 neurons of rats of different kindling, so as to provide theoretical evidence for the neuronal injury after epilepsy.DESIGN: A randomized c ntrol animal experiment.SETTINGS: Department of Neurology and Department of Anesthesiology,Qilu Hospital of Shandong University.MATERIALS: The experiments were carried out in the pathological laboratory of Shandong Academy of Medical Sciences between March and July2005. Totally 150 adult male SD rats of 260-300 g were provided by the experimental animal center of Shandong University (SCXK20030004), they were raised at room temperature and were free to the access of food and water.METHODS: The adult male Sprague Dawley (SD) rats were divided into intraperitoneal injection of kainic acid group and caudal venous injection of kainic acid group respectively ac cording to the method of random number table, and the rats were administrated by kainic acid injected intraperitoneally (12 mg/kg) and via caudal vein (10 mg/kg) respectively. Each group was divided into 5 subgroups, which were 3, 6, 24, 48 and 72 hours after status epilepticus groups respectively. Twelve successfully induced rats were selected from each subgroup, hippocampi were removed at different time points after the termination of status epilepticus, 2 were used for examination under electron microscope, 5 for the reverse transcription-polymerase chain reaction (RT-PCR) detection of Fas and Bax, and 5 for the immunohistochemical assay of Caspase-3. Another 12 rats were used as normal controls without any treatment. The materials were taken at24 hours after corresponding status epilepticus in the control group, and the specific distributions were the same as those in the subgroups. The mitochondrial structure was observed under electron microscope, the levels of Fas and Bax mRNA were detected with semi-quantitative RT-PCR, and the expression of Caspase-3 protein was determined with the immunohistochemical assay.MAIN OUTCOME MEASURES: ① Results of ultrathin section under transmission electron mcroscope; ② RT-PCR results; ③ Immunohistochemical results.RESULTS: Totally 132 rats were involved in the analysis of results. ①Mitochondrial structure under electron microscope: In the intraperitoneal injection group, the mitochondria swelled, and the neurons showed characters of apoptosis. In the caudal venous injection group, the mitochondria swelled, and accompanied by the membranous collapse, and the neurons manifested the necrosis. ② No expression of Fas and Bax was detected in the control group and caudal venous injection. In the intraperitoneal injection group, Fas expression appeared at 6 hours after status epilepticus, increased at 24 hours, reached the peak value at 48 hours, and lasted till 72 hours. ③ The Caspase-3 expressions began to increase 6 hours after status epilepticus in both the intraperitoneal injection group and caudal venous injection group(10.27±0.34, 15.21±0.34; P ＜ 0.001), and reached the peak values at 24 hours (25.36±0.47, 28.23±0.47; P ＜ 0.001); The higher expression of Caspase-3 lasted till 72 hours in the intraperitoneal injection group, but sharply decreased in the caudal venous injection group.CONCLUSION: Two different methods of administration result in different severity of mitochondrial damage and different expressions of Fas, Bax and Caspase-3, which further determines the molecular mechanisms of neuronal death.
ABSTRACT
OBJECTIVE: Epileptic attack can cause neuronal damage and increase the risk of potential seizure. Analysis of the possible mechanism of neuronal damage following epileptic seizure may provide evidences for implementing preventive measures against brain damage due to epileptic seizures.DATA SOURCES: A computer-based search of the related publications in PubMed database between June 1995 and June 2004 with different combinations of the key words of "epilepsy", "neuron damage", "necrosis"and "apoptosis", limiting the results to the language of English.STUDY SELECTION: The retrieved articles were examined at first to select reports of experimental study on human and animals related to epilepsy and the subsequent neuronal damages, and their full-text publications were obtained with the other unrelated articles excluded.DATA EXTRACTION: Eighteen articles documenting randomized controlled experiment immediately related to neuronal damage after epilepsy seizure, 4 reporting non-randomized controlled experiments related to central neuronal excitatory toxic damage, and 3 concerning neuronal damage were collected for this review.DATA SYNTHESIS: In the 14 randomized controlled experiments, chemical or electric methods were used to induce epilepsy in the animal models in which the ultrastructural changes of the neurons and cell organelles were observed and the expression of apoptosis-related factors determined.In the 4 non-randomized controlled experiments, central neuronal ischemic and hypoxic models were adopted for observing the expression of various apoptotic factors in the neurons due to different damages with the assistance of electron microscope, to provide direct evidences for the mechanism of central neuronal excitatory toxic damage. The other three related literatures introduced the pathways of neuronal damages and the expression of the related factors.CONCLUSION: Neuronal death after epileptic seizure is correlated with the severity of the damage and mitochondrial functional status, and the mitochondria constitute the control center for neuronal survival. The release of cytochrome C and the activation of caspases are the final common pathway of neuronal damage.
ABSTRACT
Objective: Serum pleiotrophin(PTN) is closely related to tumor,particularly to lung cancer.Many studies focused on PTN recently.This study is to evaluate the diagnostic value of PTN,Cyfra21-l and CEA tests independently or in a combined style in lung cancer.Methods: Sixty-one patients with lung cancer and 17 patients with benign lung lesion disease all received PTN,Cyfra21-l and CEA tests before the treatment.They were compared with pathological results,and then we calculated the diagnostic value of each method and the combined styles.Results: The sensitivity of PTN together with Cyfra21-l and CEA tests is 95.08% and it is the highest among the methods.Conclusion: PTN together with Cyfra21-l and CEA tests has a important diagnostic value through mutual complementation in detecting lung cancer.
ABSTRACT
Objective To observe the mitochondrion and nucleus ultrastructural damage and caspase-3 expression in hippocampal CA_3 neurons during kainic acid(KA) induced status epilepticus(SE) in rats.Methods SE was induced for 2 h with KA in adult male Wistar rats.3,12 and 24 h later the rats were killed and the hippocampal CA_3 subareas were taken out to make brain sections.The neuronal damage on the whole with light microscope and the ultrastructure of mitochondrion and nucleus with electron microscope.Caspase-3 expression of the same area was examined with immunohistochemical staining.Results 24 h after SE,by the light microscope examination,the KA group showed that the neurons put scattered disorder and nucleus shrink firmly.3 h after SE,electron microscope examination showed swelling cristae and ruptured membrane of mitochondria.The change of nucleus were significant margination of chromatin 24 h after SE.Compared to normal control group,the caspase-3 expression increased 12 h after SE,the average number of positive cell and the gray scale were obviously higher(all(P
ABSTRACT
0.05) . Significant decreases of contents of 14 kinds of macro-elements and trace elements in purified water were found compared with those in water samples after activated carbon filtration (t = 2.496 9-5.261 7, F2 and showed significant dose-response relationship respectively (r = 0.980, P