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Objective: To develop and validate a deep learning model based on fundus photos for the identification of coronary heart disease (CHD) and associated risk factors. Methods: Subjects aged>18 years with complete clinical examination data from 149 hospitals and medical examination centers in China were included in this retrospective study. Two radiologists, who were not aware of the study design, independently evaluated the coronary angiography images of each subject to make CHD diagnosis. A deep learning model using convolutional neural networks (CNN) was used to label the fundus images according to the presence or absence of CHD, and the model was proportionally divided into training and test sets for model training. The prediction performance of the model was evaluated in the test set using monocular and binocular fundus images respectively. Prediction efficacy of the algorithm for cardiovascular risk factors (e.g., age, systolic blood pressure, gender) and coronary events were evaluated by regression analysis using the area under the receiver operating characteristic curve (AUC) and R2 correlation coefficient. Results: The study retrospectively collected 51 765 fundus images from 25 222 subjects, including 10 255 patients with CHD, and there were 14 419 male subjects in this cohort. Of these, 46 603 fundus images from 22 701 subjects were included in the training set and 5 162 fundus images from 2 521 subjects were included in the test set. In the test set, the deep learning model could accurately predict patients' age with an R2 value of 0.931 (95%CI 0.929-0.933) for monocular photos and 0.938 (95%CI 0.936-0.940) for binocular photos. The AUC values for sex identification from single eye and binocular retinal fundus images were 0.983 (95%CI 0.982-0.984) and 0.988 (95%CI 0.987-0.989), respectively. The AUC value of the model was 0.876 (95%CI 0.874-0.877) with either monocular fundus photographs and AUC value was 0.885 (95%CI 0.884-0.888) with binocular fundus photographs to predict CHD, the sensitivity of the model was 0.894 and specificity was 0.755 with accuracy of 0.714 using binocular fundus photographs for the prediction of CHD. Conclusion: The deep learning model based on fundus photographs performs well in identifying coronary heart disease and assessing related risk factors such as age and sex.
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Humans , Male , Retrospective Studies , Deep Learning , Fundus Oculi , ROC Curve , Algorithms , Risk Factors , Coronary Disease/diagnostic imagingABSTRACT
OBJECTIVE@#To explore clinical effect of locking plate external fixation combined with membrane induction technology in treating open and comminuted tibial fractures with bone defects.@*METHODS@#Totally 92 patients of open and comminuted tibial fractures with bone defects were chosen form January 2018 to July 2019, and randomly divided into external fixation group and internal fixation group, 46 patients in each group. In external fixation group, there were 29 males and 17 females, aged from 25 to 62 years old, with an average of (37.45±10.92) years old;according to AO classification, 15 patients were type A, 22 patients were type B and 9 patients were type C;according to Gustilo classification, 21 patients were typeⅡ, 10 patients were type ⅢA, 10 patients were type ⅢB, 5 patients were type Ⅲ C;treated by fracture reduction with locking plate external fixation. In internal fixation group, there were 31 males and 15 females, aged from 23 to 60 years old, with an average of(36.88±10.64) years old;according to AO classification, 18 patients were type A, 20 patients were type B and 8 patients were type C; according to Gustilo classification, 22 patients were typeⅡ, 11 patients were type ⅢA, 7 patients were type ⅢB, 6 patients were type Ⅲ C;treated by traditional open reduction with plate internal fixation. Operation time, intraoperative blood loss, incision length, hospital stay, fracture healing time and lower limb full weight-bearing time and postoperative complications between two groups were observed and compared, bone mineral density, osteocalcin, blood calcium and phosphorus before operation and 1 month after operation.@*RESULTS@#All patients were followed up from 12 to 18 months with an average of (14.92±2.46) months. Operation time, intraoperative blood loss, incision length, hospital stay, fracture healing time and lower limb full weight-bearing time of external fixation group were significantly better than that of internal fixation group(@*CONCLUSION@#Locking plate external fixation combined with membrane induction technology in treating open and comminuted tibial fractures with severe post-traumatic bone defects has advantages of less trauma, reliable fixation, shorter fracture healing time, and could improve bone metabolic activity with less postoperative complications.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Bone Plates , External Fixators , Fracture Fixation , Fracture Fixation, Internal , Fractures, Comminuted/surgery , Technology , Tibial Fractures/surgery , Treatment OutcomeABSTRACT
OBJECTIVE@#Pulmonary nodules (PNs) are one of the imaging manifestations of early lung cancer screening, which should receive more attention. Traditional Chinese medicine believes that voice changes occur in patients with pulmonary diseases. The purpose of this study is to explore the differences in phonetic characteristics between patients with PNs and able-bodied persons.@*METHODS@#This study explores the phonetic characteristics of patients with PNs in order to provide a simpler and cheaper method for PN screening. It is a case-control study to explore the differences in phonetic characteristics between individuals with and without PNs. This study performed non-parametric statistics on acoustic parameters of vocalizations, collected from January 2017 to March 2018 in Shanghai, China, from these two groups; it explores the differences in third and fourth acoustic parameters between patients with PNs and a normal control group. At the same time, computed tomography (CT) scans, course of disease, combined disease and other risk factors of the patients were collected in the form of questionnaire. According to the grouping of risk factors, the phonetic characteristics of the patients with PNs were analyzed.@*RESULTS@#This study was comprised of 200 patients with PNs, as confirmed by CT, and 86 healthy people that served as a control group. Among patients with PNs, 43% had ground glass opacity, 32% had nodules with a diameter ≥ 8 mm, 19% had a history of smoking and 31% had hyperlipidemia. Compared with the normal group, there were statistically significant differences in pitch, intensity and shimmer in patients with PNs. Among patients with PNs, patients with diameters ≥ 8 mm had a significantly higher third formant. There was a significant difference in intensity, fourth formant and harmonics-to-noise ratio (HNR) between smoking and non-smoking patients. Compared with non-hyperlipidemia patients, the pitch, jitter and shimmer of patients with PNs and hyperlipidemia were higher and the HNR was lower; these differences were statistically significant.@*CONCLUSION@#This measurable changes in vocalizations can be in patients with PNs. Patients with PNs had lower and weaker voices. The size of PNs had an effect on the phonetic formant. Smoking may contribute to damage to the voice and formant changes. Voice damage is more pronounced in individuals who have PNs accompanied by hyperlipidemia.
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BACKGROUND@#Malignant pleural effusion (MPE) is a complicated condition of patients with advanced tumors. Further dissecting the microenvironment of infiltrated immune cells and malignant cells are warranted to understand the immune-evasion mechanisms of tumor development and progression.@*METHODS@#The possible involvement of microRNAs (miRNAs) in malignant pleural fluid was investigated using small RNA sequencing. Regulatory T cell (Treg) markers (CD4, CD25, forkhead box P3), and Helios (also known as IKAROS Family Zinc Finger 2 [IKZF2]) were detected using flow cytometry. The expression levels of IKZF2 and miR-4772-3p were measured using quantitative real-time reverse transcription polymerase chain reaction. The interaction between miR-4772-3p and Helios was determined using dual-luciferase reporter assays. The effects of miR-4772-3p on Helios expression were evaluated using an in vitro system. Correlation assays between miR-4772-3p and functional molecules of Tregs were performed.@*RESULTS@#Compared with non-malignant controls, patients with non-small cell lung cancer had an increased Tregs frequency with Helios expression in the MPE and peripheral blood mononuclear cells. The verified downregulation of miR-4772-3p was inversely related to the Helios Tregs frequency and Helios expression in the MPE. Overexpression of miR-4772-3p could inhibit Helios expression in in vitro experiments. However, ectopic expression of Helios in induced Tregs reversed the effects induced by miR-4772-3p overexpression. Additionally, miR-4772-3p could regulate Helios expression by directly targeting IKZF2 mRNA.@*CONCLUSION@#Downregulation of miR-4772-3p, by targeting Helios, contributes to enhanced Tregs activities in the MPE microenvironment.
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Background@#Malignant pleural effusion (MPE) is a complicated condition of patients with advanced tumors. Further dissecting the microenvironment of infiltrated immune cells and malignant cells are warranted to understand the immune-evasion mechanisms of tumor development and progression.@*Methods@#The possible involvement of microRNAs (miRNAs) in malignant pleural fluid was investigated using small RNA sequencing. Regulatory T cell (Treg) markers (CD4, CD25, forkhead box P3), and Helios (also known as IKAROS Family Zinc Finger 2 [IKZF2]) were detected using flow cytometry. The expression levels of IKZF2 and miR-4772-3p were measured using quantitative real-time reverse transcription polymerase chain reaction. The interaction between miR-4772-3p and Helios was determined using dual-luciferase reporter assays. The effects of miR-4772-3p on Helios expression were evaluated using an in vitro system. Correlation assays between miR-4772-3p and functional molecules of Tregs were performed.@*Results@#Compared with non-malignant controls, patients with non-small cell lung cancer had an increased Tregs frequency with Helios expression in the MPE and peripheral blood mononuclear cells. The verified downregulation of miR-4772-3p was inversely related to the Helios+ Tregs frequency and Helios expression in the MPE. Overexpression of miR-4772-3p could inhibit Helios expression in in vitro experiments. However, ectopic expression of Helios in induced Tregs reversed the effects induced by miR-4772-3p overexpression. Additionally, miR-4772-3p could regulate Helios expression by directly targeting IKZF2 mRNA.@*Conclusion@#Downregulation of miR-4772-3p, by targeting Helios, contributes to enhanced Tregs activities in the MPE microenvironment.
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During the uric acid production, excretion and reabsorption in the liver, kidney and intestine, several uric acid transporter proteins are involved in these processes. A large number of studies have shown that glucose transporter 9 plays an important role in the uric acid transport in the liver, kidney and intestine, and participates in the uric acid reabsorption. The ATP-binding cassette superfamily G member 2 is mainly expressed in the apical membrane of the proximal tubular epithelial cells of the kidney, which is involved in the uric acid secretion. The multidrug resistant protein 4 is expressed in the apical membrane of the renal tubular epithelial cells, which transfers uric acid from the renal tubular epithelial cells into the renal tubular lumen. The urate-anion transporter 1 as well as the organic anion transporters 1 and 3 are all the organic anion transporters belonging to the SLC22 A family of transmembrane transporters, and all participate in the uric acid transport in the kidney, especially the uric acid secretion and excretion. In this review, we summarize the research progress of these uric acid transporters, focusing on their effects on the regulation of the serum uric acid balance.
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In this paper, on the contrast of healthy leech, the bacterial diversities were analyzed by 16S rDNA sequence analysis of the bacteria of muscle and intestinal tract of Whitmania pigra, the environment water and sediment of cultivating the diseased Wh. pigra in high temperature by high-throughput sequencing to determine the possible pathogenic bacteria of bacterial diseases of Leech in high temperature. The results showed that the original sequence reached over 83 000, and the effective sequences accounted for more than 87%. The GC contents ranged from 52% to 54% and the bacterial diversities were abundant. Bacterial relative abundance analysis showed that the bacterial content of Proteobacteria, Bacteroidetes, and Firmicutes was the most abundant in all treatments. Compared with healthy leech muscles and intestines, the muscle and intestinal tract of pathogenic leech relative abundance of Bacteroides, Pseudomonas, and Desulfovibrio was significantly increased, and it was abundant in water and sediment of diseased leeches, Lead to the possibility that the pathogenic bacteria of this bacterial disease may be Bacteroides, Pseudomonas, Desulfovibrio.
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Objective To compare the curative effect of open and minimally invasive treatment of acute closed achilles tendon rupture with 5 years followed-up study.Methods From September,2010 to January,2012,28 patients with acute closed Achilles tendon rupture in our hospital were followed up for 5 years.There were 21 males and 7 females.Minimally invasive percutaneous suture in 11 cases;open suture in the treatment of 17 cases.The patients were followed up at 6 months,1 year,2 years,3 years,4 years,and 5 years after AOFAS and ATRS score.Results The follow-up time ranged from 60 to 72 months.All incisions healed by first intention and no incision related complications occurred.2 groups of patients with ATRS score at 6 months after operation:the open group was 81.23±3.99,minimally invasive percutaneous group of 88.27±4.27,the difference between the two groups was statistically significantly.After 1 year,there was no significant difference in the scores between the two groups.Two groups of patients with AOFAS score at 6 months after operation:the open group was 69.00±6.23,minimally invasive percutaneous group of 79.27±4.83,the difference between the two groups was statistically significantly.At one year after operation,the open group was 85.53±3.38,and the minimally invasive group was more than 89.90±3.38.The difference between the two groups was statistically significantly.After 2 years,there was no significant difference in the scores between the two groups.Conclusion There is no significant difference between the 2 years after surgery in the treatment of acute closed Achilles tendon rupture or open surger.
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Objective:To investigate the effect of Nutrison MCT and Nutrison MF enteral nutrition on gut barrier function during the perioperative period in patient with malignant obstructive jaundice.Methods:82 cases of patients with malignant obstructive jaundice were randomized divided into Nutrison MCT group with 40 cases and Nutrison MF group with 42 cases.Enteral nutrition was given on admission.The level of urinary lactulose and mannitol(L/M) ratio,plasma endotoxin and diamine oxidase(DAO) activity on the 2nd day after admission,lday before surgery,5 days after full dose of enteral nutrition were collected.Results:Before surgery,no statistical significance difference of urinary L/M ratio,plasma endotoxin and DAO activity were found between the 2nd day after admission and 1day before surgery.After full dosing of enteral nutrition,urinary L/M ratio,plasma endotoxin and DAO activity were decreased,while Nutrison MCT group decreased more significantly than Nutrison MF group.Conclusion:Nutrison MCT can improve the gut barrier function of patients with malignant obstructive jaundice more earlier than Nutrison MF after surgery.
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Objective To observe the thickness and histologic changes of cartilage -perichondrium complex (CP) and vascular density changes in perichondrium after repairing perforated eardrum .Methods Forty -five male guinea pigs were randomly divided into three groups ,and were used to create left TM perforation .The left cavum conchae cartilage-perichondrium was immediately used to repair the perforation .The observation time for group A was two weeks ,group B was six weeks and group C was twelve weeks .The thickness of the composite was meas-ured before and after the implantation .The histologic changes of implants and the vascular change in perichondri were observed .Results The healing time for the left ear was 3 .8 ± 0 .84 ,and the right ear 7 .2 ± 0 .84 .The thick-ness of the composite for group A increased by 14 .4% (P0 .05) .Group C decrease by 14 .2% (P<0 .05) .After implantation ,the thickness of cartilage in group C was decreased by 34 .2% compared to that in group A (P<0 .05) .At the 2nd week in post -implantation ,the number of vessels within the unit area of fiber -perichondrium was up to 13 .28 ± 2 .49 ,while the above number reached to 7 .71 ± 2 .49 at the 6th week after implantation .The final histological structure of recon-structed tympanic membrane at the 2nd week and 6th week was composed of squamous epithelial layer ,fibrous -perichondrium layer ,cartilaginous layer and mucous layer .The above histological structure at 12th week was com-posed of squamous epithelial layer ,cartilaginous layer and mucous layer .Conclusion The healing time of the recon-structed tympanic membrane is shorter than the time spent on healing on its own .The thickness of the composite in post-implantation in three groups first increased and then decreased compared to that in pre -implantation .The number of blood vessels of fiber -perichondrium in the reconstructed tympanic membrane first increased and then re-duced .The layers of final histological structure of reconstructed tympanic membrane are similar to those of normal tympanic membrane .From the perspective of histology ,the cartilage-perichondrium composite is the suitable ma-terial for repairing tympanic membrane .
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Objective To evaluate the efficacy of 9 μ g tiotropium inhalation once daily in patients with moderate to severe chronic obstructive pulmonary disease(COPD).Methods One hundred seventy-one patients with moderate to severe COPD were divided into two groups by random digits table methods:86 patients in test group who used 9 μ g tiotropium powder inhaler once daily,and 85 patients in control group who used the recommended dose 18 μ g once daily,both for 6 months.Patients' pulmonary function and 6-minute walk test were measured before treatment,3 months after treatment and 6 months after treatment.Results Two groups were compared 6 months after treatment.The level of forced expiratory volume in first second(FEV1) in the control group increased an average of (0.21 ± 0.12) L,and the rising rate was (19.31 ± 10.61)％.The level of FEV1 in test group increased (0.22 ±0.13) L,and the rising rate was (19.25 ± 11.52)％.The level of forced vital capacity (FVC) in control group increased an average of (0.22 ±0.10) L,and the rising rate was (11.63 ±5.31)％.The level of FVC in test group increased (0.23 ± 0.15) L,and the rising rate was (11.62 ± 6.45)％.The distance of 6-minute walk test in control group increased an average of (29.24 ± 11.92) m after treatment for 6 months,and the rising rate was (18.30 ± 3.77)％.The distance of 6-minute walk test in test group increased an average of (29.14 ± 11.89) m,and the rising rate was (18.20 ± 3.85)％.The number of acute exacerbations and rehospitalizations in two groups at these period had no significant difference (P ＞ 0.05).As for the test group,though the number of acute exacerbations and rehospitalizations decreased,there were no significant differences (P ＞ 0.05).Conclusions The effect of inhalation therapy of 9 μ g tiotropium once daily is more or less the same with that of 18 μ g once daily.
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BACKGROUND:PA6 cells are bone marrow stromal stem cells from the mouse skul , and scientists have found that PA6 cells co-cultured with some kinds of stem cells have shown neural differentiation, based on which, PA6 cells can be used for repair of nerve injury. Therefore, researchers give more and more attentions to PA6, but few studies have addressed isolation and culture methods of bone marrow stromal stem cells from the skul . OBJECTIVE:To isolate and culture bone marrow stromal stem cells from the skul of Sprague-Dawley rats and to observe cellular morpholopy in vitro and perform immunofluorescence identification. METHODS:Under sterile conditions, newborn Sprague-Dawley rat’s skul was cut into pieces. Smal skul pieces were washed using Dulbecco’s modified Eagle’s medium containing 10%fetal bovine serum. Single cellsuspension was made, and placed in culture flask. The culture medium was changed many times for cellpurification. The second passage of cells was obtained for morphology observation under an inverted microscope. cellsurface markers were detected by using immunofluorescence staining. RESULTS AND CONCLUSION:After the primary culture for 24 hours, cells exhibited adherent growth;after 3 days, cells were increased in number, and presented with irregular shapes, such as polygon, triangle, spindle and flat shape. After passage, cells were uniform in morphology, arranged in radial and bunch patterns, and exhibited strong adherent ability. Some cells grew in cluster, and proliferated faster than primary cells. Immunofluorescence staining showed that cells were positive for CD105, CD73, CD44, CD90, but negative for CD45, D34,CD14, HLA-DR. Results indicate that this method can obtain bone marrow stromal stem cells.
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ObjectiveTo evaluate the effectiveness of treating tuberculous pleurisy with hydrogen peroxide solution washing and povidone iodine solution perfusing topically.MethodsAfter fixing position with B ultrasound or 64 rows spiral computed tomography,the thoracic cavity was firstly inspected with thoracoscope,the effusion was exhausted and the conglutionations was cut with thoracoscope or high frequency electrome,then the thoracic cavity was washed with 200 ml 3％ hydrogen peroxide solution,and lastly it was perfused continually with 0.5％ povidone iodine solution to control the intracavity infection quickly and to promote cicatrization.After routine antitubercular therapy for one year,the time of pleural effusion disappearing and the state of pleura hypertrophy was observed.ResultsThere were 86 cases in all.After antitubercular therapy and extracting pleural effusion at the same time in 48 cases.Twenty-two cases'effusion didn't increase basically after 4 weeks.They stopped extracting effusion and went on with antitubercular therapy (the first group).The rest 26 patients continued to accept the therapy of hydrogen peroxide solution washing and povidone iodine solution perfusing topically(the second group),and 23cases achieved remedial goal in a week with the effective rate of 88.46％ (23/26).The third group had 38 cases who didn't accept thoracocentesis but accepted the therapy of hydrogen peroxide solution washing and povidone iodine solution perfusing directly,and there were 36 cases that achieved remedial goal in a week with effective rate of 94.74％ (36/38).The time of pleural effusion disappearing in the third group was obviously shorter than that in the first group (1 week vs.4 weeks )(P ＜ 0.05 ),and there was no significant difference in the efftective rate between the second group and the third group (P ＞ 0.05 ).Half a year after therapy,the thickness of pleura was measured and it was found that in the first group was thickest and in the third group thinnest and even some without hypertrophy.It was the same effect after one year.Conclusions The effectiveness of the method treating tuberculous pleurisy with hydrogen peroxide solution washing and povidone iodine solution perfusing topically is defimite,and the method can shorten the time of pleural effusion disappearing and alleviate the pleural hypertrophy.
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<p><b>OBJECTIVE</b>To explore whether there are gene mutations of Tolloid-like 1 (TLL-1) gene in Chinese patients with sporadic congenital heart disease (CHD).</p><p><b>METHODS</b>One hundred and fifteen patients with sporadic CHD were selected as CHD group. One hundred and two age and gender-matched healthy people were recruited as control group. After amplifying the exon 10 of the TLL-1 gene by polymerase chain reaction, the polymerase chain reaction products were purified, sequenced and analyzed in order to investigate the TLL-1 gene mutation.</p><p><b>RESULTS</b>An insertion mutation of base A in the exon 10 of TLL-1 gene was identified in 7 out of 115 CHD patients, including 3 patients with atrial septal defect, 2 patients with ventricular septal defect, 1 patients with patent ductus arteriosus and 1 patients with complex CHD, the total mutation rate was 6.1% in CHD group and 0 in control group (P < 0.01).</p><p><b>CONCLUSIONS</b>TLL-1 gene mutation with an insertion mutation of base A in exon 10 is often in Chinese patients with various CHD. The underlying pathogenesis between TLL-1 gene mutation and occurrence of congenital heart disease in Chinese people remains unclear and warrants further investigations.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , Asian People , Genetics , Base Sequence , Case-Control Studies , Exons , Heart Defects, Congenital , Genetics , Mutagenesis, Insertional , Pedigree , Tolloid-Like Metalloproteinases , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of a novel metastasis-inducing protein human anterior gradient-2 (AGR2) in breast cancer and its clinical and prognostic significance.</p><p><b>METHODS</b>AGR2 expression was assessed in 160 cases of breast cancer and 20 cases of benign breast diseases by immunohistochemistry using tissue chip technology. In addition the expression of ERa, PR and c-erbB-2 in breast cancer was also evaluated. Follow-up information of 5-year duration was available in 127 patients with breast cancer. Kaplan-Meier analysis and COX regression model were used to analyze the correlation between AGR2 expression and the follow-up clinical data.</p><p><b>RESULTS</b>The expression of AGR2 was significantly higher in breast cancers than that in benign diseases (68.3% vs. 25.0% , P < 0.01). There was a negative correlation between AGR2 expression and the histological grade of breast cancer (P <0.05) , whereas positive correlations was found between the expression of AGR2 and ERalpha (P <0.05), and between the expression of AGR2 and PR (P <0.01). In the subgroup of ERalpha-positive breast cancer, Logistic regression model demonstrated AGR2 and TNM stage were important factors affecting lymph node metastasis (both P < 0.01). Kaplan-Meier analysis demonstrated that a positive expression of AGR2 was associated with poor overall survival and relapse-free survival (both P <0.01). Moreover, COX regression model confirmed the expression of AGR2 as an independent prognostic factor among patients with ERa-positive breast cancer (P <0.01).</p><p><b>CONCLUSIONS</b>The abnormal expression of AGR2 may play a role in the pathogenesis and progression of breast cancer. The metastasis-inducing capability of AGR2 may be partly regulated through the ER pathway. Therefore, AGR2 may be a useful molecular marker for prognostication for patient with hormone-responsive breast cancer.</p>
Subject(s)
Female , Humans , Antineoplastic Agents, Hormonal , BRCA2 Protein , Genetics , Metabolism , Biomarkers, Tumor , Breast Neoplasms , Diagnosis , Genetics , Metabolism , Estrogen Receptor alpha , Metabolism , Gene Expression Regulation, Neoplastic , Genetics , Immunohistochemistry , Neoplasm Metastasis , Diagnosis , Neoplasm Staging , Prognosis , Proteins , Genetics , Metabolism , Receptor, ErbB-2 , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of carbon disulfide (CS2) on inducible nitric oxide synthase (iNOS) and apoptosis in in the retina of rats.</p><p><b>METHODS</b>Twenty-four male SD rats were randomly divided into 3 groups the control group, the group receiving high-dose CS2 and the group receiving low-dose CS2. After treatment, retina were harvested and made into slice. The thickness of inner retina including outer plexiform layer, inner nuclear layer, ganglion cell layer, optic nerve fiber and inner limiting membrane was measured. Expression of iNOS in retina was measured by NADPH-NDP histochemical assay. Apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL).</p><p><b>RESULTS</b>(1) The thickness of inner retina decreased in groups receiving CS2. There was significant difference between control group and groups receiving CS2 (P < 0.05). There was also significant difference between group receiving high-dose CS2 and group receiving low-dose CS2 (P < 0.05). (2) The expression of iNOS increased in groups receiving CS2. There was significant difference between control group and groups receiving CS2 (P < 0.05 or P < 0.01). There was also significant difference between the group receiving high-dose CS2 and the group receiving low-dose CS2 (P < 0.05). (3) Apoptosis was observed in groups receiving CS2. There were significant differences in apoptosis index between the control group and groups receiving CS2 (P < 0.05 or P < 0.01). There was also significant difference between the group receiving high-dose CS2 and the group receiving low-dose CS2 (P < 0.05).</p><p><b>CONCLUSIONS</b>CS2 can evoke the expression of iNOS, and the nitric oxide thus produced can be one of the causes of retina destruction. And apoptosis is also one of the causes of retina destruction.</p>
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Animals , Male , Rats , Apoptosis , Carbon Disulfide , Toxicity , Nitric Oxide Synthase Type II , Metabolism , Rats, Sprague-Dawley , Retina , PathologyABSTRACT
To study the function of the GnRH protein, the recombinant pMAL-GnRH was constructed and expressed in TB1 E. coli. The cDNA encoding gonadotropin-releasing hormone (GnRH) and GnRH associated peptide (GAP) was amplified from total RNA of O. aurea pituitary glands by reverse transcription polymerase chain reaction (RT-PCR), and then blasted against other GnRH cDNA sequences in the GenBank. The analysis of the sequence data indicated that the coding region of the cDNA fragment, which encoded 89 amino acid residues, was about 400 bp in size. The amplified cDNA fragment was cloned into the prokaryotic expression vector, pMAL-c2x, to produce the expression vector pMAL-GnRH. The recombinant plasmid was transformed into E. coli TB1. GnRH-MBP fusion protein was obtained after the addition of IPTG into the growth media. SDS-PAGE analysis revealed that the GnRH-MBP was expressed after induction with IPTG for 4 h. A protein band of 56 kD appeared on SDS-PAGE gel and was proved by Western blot. The mass production of the recombinant protein was about 41.6% of total bacteria protein. After purification and cleavage of the fusion protein purified GnRH protein could be obtained. Then the fusion protein was used to immunise some ICR mice to produce anti-GnRH antibody. This fusion protein could significantly elicit specific antibody response in immunized mice compared with the blank groups, and the titers against GnRH reached peak 0.707 +/- 0.320 at the 5th week after immunization. These results demonstrated that recombinant protein could induce high GnRH antibody responses in laboratory animals.
Subject(s)
Animals , Mice , Cloning, Molecular , Escherichia coli , Genetics , Gonadotropin-Releasing Hormone , Genetics , Allergy and Immunology , Physiology , Mice, Inbred ICR , Plasmids , Recombinant Fusion Proteins , Allergy and Immunology , Reverse Transcriptase Polymerase Chain Reaction , Tilapia , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of uncoupling protein 2 (UCP2) and its relationship to the content of adenosine triphosphate (ATP) in livers of nonalcoholic fatty liver disease (NAFLD) rats fed a fat-rich diet.</p><p><b>METHODS</b>To produce a NAFLD model, a fat-rich diet, consisting of 10% lard oil + 2% cholesterol, was given to Sprague-Dawley rats for a period of 8, 12, 16 and 24 weeks. The normal control rats were fed normal diets. The expressions of UCP2 in the liver were detected by immunohistochemistry and semi-quantitative RT-PCR. The content of ATP of liver was measured by fluorometry.</p><p><b>RESULTS</b>Simple fatty livers were observed in the model group after 8 weeks. From 12 week to 24 week, the livers of the model group rats gradually progressed from simple steatohepatitis to steatohepatitis with pericellular fibrosis. Both immunohistochemistry and semi-quantitive RT-PCR suggested the up-regulated expression of UCP2 in these NAFLD rat livers. The hepatic expression of UCP2 mRNA in the model group was increased with time, and peaked in 24 week by 4.2 times compared to the control group ( t = 16.474, P < 0.01). The ATP content of livers was significantly reduced in the model group compared with the control group at 16 weeks [(2.97+/-0.48) x 10(-8) micromol/g vs. (2.25+/-0.55) x 10(-8) micromol/g, t = 2.419, P < 0.05] and 24 weeks [(2.97+/-0.48) x 10(-8) micromol/g vs. (1.99+/-0.66) x 10(-8) micromol/g, t = 3.248, P < 0.01]. Furthermore, there was a negative correlation between the UCP2 mRNA expression and the content of ATP in the livers of the NAFLD group (r = -0.93, P < 0.01).</p><p><b>CONCLUSIONS</b>The rat model of NAFLD could be replicated sucessfully by feeding a fat-rich diet for 24 weeks, and the mRNA and its protein of UCP2 were expressed un-regulated in livers of NAFLD. The increasing UCP2 might play a role in the reduction of ATP content in livers of the NAFLD rats.</p>
Subject(s)
Animals , Male , Rats , Adenosine Triphosphate , Metabolism , Dietary Fats , Fatty Liver , Metabolism , Ion Channels , Genetics , Liver , Metabolism , Mitochondrial Proteins , Genetics , Random Allocation , Rats, Sprague-Dawley , Uncoupling Protein 2ABSTRACT
<p><b>OBJECTIVE</b>To investigate the dynamic changes of plasma levels of prostacycline (PGI2) and thromboxane A2 (TXA2) and their relationship with the severity of hepatic injury in rats with nonalcoholic fatty liver disease (NAFLD).</p><p><b>METHODS</b>We established a NAFLD model, with a fat-rich diet consisting of 10% lard oil + 2% cholesterol, which was given to Sprague-Dawley rats (n=48) for a period of 8, 12, 16 and 24 weeks. The other rats were fed standard diets and were used as normal controls (n=24). At sacrifice, liver pathology scores were evaluated and plasma levels of PGI2, its stable metabolic product 6-keto-PGF1 alpha and TXA2, and TXB2 were determined by radioimmunoassay.</p><p><b>RESULTS</b>Simple fatty livers were observed in the model group at 8 weeks. From 12 weeks to 24 weeks, the livers gradually progressed from simple steatohepatitis to liver fibrosis. Plasma levels of TXB2 in the model group increased higher than in the control group after 8 weeks [(52.4+/-3.15) ng/L vs (41.1+/-1.45) ng/L] and continued to increase over time, with the highest levels at 24 weeks [(117.7+/-7.47) ng/L]. A strong positive correlation (r=0.537) was seen between plasma TXB2 levels and the severity of liver injury. Plasma 6-keto-PGF1 alpha concentrations decreased in the model group in comparison with the control group after 8 weeks [(31.1+/-1.62) ng/L vs (36.5+/-1.68) ng/L] and continued to decrease over time, with the lowest concentrations at 24 weeks [(3.4+/-2.43) ng/L t=3.77]. A negative correlation was shown between the 6-keto-PGF1 alpha level and the severity of the liver injury.</p><p><b>CONCLUSION</b>A rat model of NAFLD was established successfully by feeding a fat-rich diet for 24 weeks. In this model, the imbalance of plasma PGI2 and TXA2 levels (increased TXB2 and decreased 6-keto-PGF1 alpha levels) may play a role in the pathogenesis of experimental NAFLD.</p>
Subject(s)
Animals , Male , Rats , 6-Ketoprostaglandin F1 alpha , Blood , Epoprostenol , Blood , Fatty Liver , Blood , Liver , Pathology , Rats, Sprague-Dawley , Thromboxane A2 , Blood , Thromboxane B2 , BloodABSTRACT
<p><b>OBJECTIVE</b>To explore the role of endotoxin in the pathogenesis of nonalcoholic steatohepatitis (NASH).</p><p><b>METHODS</b>Rat models of NASH were established by giving a fat-riched diet. These rats were sacrificed at the 4th, 8th, 12th, 16th and 24th weeks during the study. The other rats fed with normal diet were taken as normal controls at the same stage during the study. The blood of abdominal aorta was obtained and the levels of serum endotoxin, tumor necrosis factor-alpha (TNF-a), and interleukin-1 beta (IL-1 b) were measured. The expression of CD(14) and lysozyme in rats' livers were detected by immunohistochemistry.</p><p><b>RESULTS</b>Rat models of NASH with liver fibrosis were established successfully. The levels of endotoxin in aorta blood of NASH rats increased significantly at the 24th week (0.23 EU/L 0.06 EU/L vs 0.15 EU/L 0.03 EU/L, t>2.179, p <0.05) while the expression of CD(14) increased from the 4th week, and the Kupffer cells expressing lysozyme were activated, then kept increasing activation through the study. In NASH rats, the levels of serum TNF-a increased from the 8th week (26.39 pg/ml 24.21 pg/ml vs 9.82 pg/ml 9.29 pg/ml, t>2.145, p < 0.05) and serum IL-1beta increased from the 16th week (23.76 pg/ml 21.81 pg/ml vs 6.25 pg/ml 2.98 pg/ml, t>2.145, p<0.05).</p><p><b>CONCLUSION</b>Liver injury results from endotoxin existing in NASH rats which may play an important role in the pathogenesis of NASH by activating Kupffer cells and inducing the production of cytokines, such as TNF-a.</p>