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Objective To investigate the gene expression of sigma factors in vivo, and to explore the sigma factors that may be closely related to the virulence of pathogenic Mycobacterium tuberculosis Methods Tuberculosis (TB) patients diagnosed in the outpatient department of Tianjin Tuberculosis Control Center from January to December 2018 were selected, and 20 sputum-positive specimens were randomly selected from TB patients confirmed with Xpert-positive for the present study. Two immediate sputum specimens were collected from each case of pulmonary tuberculosis before treatment, one for RNA extraction and one for in vitro culture. In vitro cultured strains in the logarithmic phase of growth were harvested for RNA extraction. The specific primers for 13 sigma factors were designed. The differential expression of the 13 sigma factors between sputum isolates and in vitro cultured strains was analyzed by fluorescence quantitative PCR. Taking ribosomal 16s as the reference gene, the transcription level of sigma factors was analyzed by 2ΔCt. Using the stably expressed sigA as the control reference, the expression differences of other sigma factors were analyzed by one-way ANOVA. Results Within 0 days, stress-associated sigma factors have a different expression profile in clinical isolate strains vs H37Rv or in vitro. All the sigma factors induced up regulation in sputum ,while no difference transcription between clinical isolate strains vs H37Rv（P＞0.05）. When compared to in vitro culture ,only sigM transcript highest in sputum(P＜0.05). Conclusion SigM plays an important role in the initial stages of bacterial infection, but its exact role is unclear.We assumed it could have a role in the interplay between the host immune defenses and the bacterial escape mechanisms.
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Chronic hepatitis B virus infection (CBI) and nonalcoholic fatty liver disease (NAFLD) are the two main etiologies of chronic liver diseases worldwide, and therefore, concomitant CBI and NAFLD (Co-CBI&NAFLD) is relatively common. There are still controversies over the influence of the interaction between CBI and NAFLD, especially NAFLD, on the progression, antiviral response, and outcome of chronic hepatitis B (CHB). Current clinical and basic research on Co-CBI&NAFLD have shown that NAFLD could inhibit HBV DNA replication to a certain degree, manifesting as a relatively low HBV DNA load, and it might increase HBsAg clearance rate in patients with CHB. Prospective or retrospective cohort studies have shown that patients with Co-CBI&NAFLD tend to have more rapid progression of liver fibrosis than those with CHB alone, as well as increased incidence rates of liver cirrhosis and hepatocellular carcinoma. Histologically, Co-CBI&NAFLD has the pathological changes of both CHB and NAFLD, and therefore, it is difficult to identify their own characteristic lesions. At present, the natural history and pathogenesis of Co-CBI&NAFLD remain unclear, and its pathological characteristics have not been fully identified. There is still a lack of high-level evidence-based supporting information on the influence of NAFLD on the course of CHB, especially its impact on antiviral response and disease outcome, and there are also no guidelines for the diagnosis/treatment or management of Co-CBI&NAFLD in China and globally. Solutions to the above issues will definitely deepen the understanding of Co-CBI&NAFLD, standardize and improve clinical diagnosis/treatment or management, and thus reduce the incidence and mortality rates of related end-stage liver diseases.
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Objective To evaluate the performance of GenoType®MTBDRplus VER2.0 kit for detecting Mycobacterium tuberculosis complex (MTBC) in sputum. Methods Sputum samples from 177 patients with suspected tuberculosis were collected, and tested by smear, MGIT liquid culture and GenoType® MTBDRplus VER2.0. When the liquid culture was positive, identification of strains was carried out. The results were statistically analyzed using SPSS 22.0, and the consistency of the count data was compared using the Kappa test. Good consistency was defined as K≥0.75. The sensitivity and specificity were applied to evaluate the performance of GenoType® MTBDRplus VER2.0 kit for detecting MTBC in sputum. Results Based on the MGIT liquid culture results,，the sensitivity, specificity, positive predictive value, negative predictive value and total coincidence rate of the GenoType® MTBDRplus VER2.0 kit was 91.67%, 95.58%, 91.67%, 95.58% and 94.22%，respectively．The Kappa test was performed on both methods, K=0.872 (P® MTBDRplus VER2.0 kit had high sensitivity and specificity for detecting MTBC in sputum, and it has good application value for early diagnosis of tuberculosis.
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This article reported a case of malignant pheochromocytoma and paraganglioma with cranial metastasis. The patient was a 25-year-old man. At the age of 10, he was diagnosed with retroperitoneal paraganglioma. He received radiotherapy after partial resection of the tumor. At the age of 15, when he consulted for headache and hypertension, a space occupying lesion was found on the right adrenal without further diagnosis and treatment. His headache and hypertension were aggravated at the age of 25. Then he visited our medical center and diagnosed with right adrenal pheochromocytoma. He undertook an operation to remove the right adrenal lesion, and his blood pressured could be controlled in the normal range with oral antihypertensive medication. One year after the surgery, a cranial lesion with positive 131I-metaiodobenzylguanidine imaging was found following the increase of norepinephrine and dopamine in both blood and urine. The diagnosis of malignant pheochromocytoma with cranial metastasis was confirmed. The cranial lesion was treated with cyberknife therapy for 4 times, and the lesion was slightly reduced. Four year after the removal of right adrenal tumor, he developed multiple bone metastasis in the right ribs and left ilium, and then he received 131I-metaiodobenzylguanidine radiotherapy. Additionally, the results of gene test indicated a pathogenic mutation of SDHB. During the follow-up period of one year after the treatment, his blood pressure fluctuated to about 140/90mmHg with 10mg of phenoxybenzamine once a day. The case is a malignant pheochromocytoma with multiple bone metastasis characterized by a long course with an ideal prognosis in a way. Consequently, we suggested the diagnostic pathway with genetic testing based on clinical phenotype to approach more timely diagnosis and reasonable evaluation of the prognosis in pheochromocytoma and paraganglioma. Furthermore, clinicians should pay more attention to the long-term surveillance.
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Objective@#To investigate the relationship between after-school schedule and bone mineral density in middle-school students in Shanghai, to provide a reference for taking targeted measures.@*Methods@#From November 2017 to April 2018，eighteen classes of six middle schools in Shanghai (from urban districts, urban-suburb combined districts and suburban districts, respectively) were selected based on cluster random sampling. In each school, 2-4 classes were further randomly chosen in the 6th grade. Questionnaires were completed by 518 students and their parents, as well as physical examination and bone mineral density assessment.@*Results@#The average time spent on after-school academic learning during weekends was 4.0 (IQR: 2.0, 6.0) h. The Z-score of bone density was 0.3 (IQR: -0.7, 1.0). The smooth curve fit model showed a non-linear relationship between after-school academic learning time during weekends and the Z-scores of bone mineral densities. A two-stage multiple linear regression analysis was further applied according to the fit results, and the results showed that when total afterschool academic learning time <4.5 hours during weekends, the learning time was inversely correlated with the bone density Z-scores (β=-0.11,P=0.01), and when the learning time ≥4.5 hours, there was no significant correlation between the learning time and bone density Z-scores (β=0.02, P=0.65). Parent and student questionnaires showed that there was a non-linear relationship between students’ daily time spent on outdoor sports-related activities and bone density Z-scores. When time spent on the outdoor activities ≥45 minutes per day, outdoor activities were positively correlated with bone density Z-scores (P<0.05). However, when the time spent on outdoor activities <45 minutes per day, there was no significant relationship between outdoor activities and bone density Z-scores (P>0.05).@*Conclusion@#Too much time on after-school academic learning during weekends or limited time on outdoor activities are both related to impaired bone mineral densities. Therefore, a reasonable after-school schedule for middle-school students is important for physical development, especially during weekends.
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Objective To determine the correlation between papillary thyroid cancer( PTC) malignancy with thyroid antibody expression level. Methods In this retrospective study, 4046 goiter patients with complete clinical data who have undergone thyroidectomy were separated into the benign nodule ( n=1357) and PTC groups ( n=2689), and their preoperative thyroglobulin antibody ( TgAb) and thyroid peroxidase antibody ( TPOAb) were assessed through antibody analysis. HT was determined based on the postoperative histopathology. The collected data were then evaluated for the correlation with PTC malignancy through statistical tests. Results In general, patients in the PTC group showed a significantly higher TgAb positivity ( 10. 9％) than those in the benign group ( 5. 6％, P<0.01) . Among female patients, a higher TgAb positivity was observed for those in the PTC group than those in the benign group (14.0％ vs 7.0％, P<0.01), while no significant difference between groups was observed in male patients. The high odds ratio of 2.18 supports the significant correlation between TgAb-positive status and higher risk of developing PTC in women (P<0.05). However, the risk of developing PTC was not correlated with higher TgAb titre. TgAb was not significantly correlated with the mass size, multifocality, lymph node metastasis or the TNM classification of malignant tumor stage Ⅲ/Ⅳ. Conclusion The increasing risk of PTC is related to TgAb among female patients, but it will not increase as TgAb titer rises. It is not associated with PTC aggressiveness.
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Objective To explore the clinical and MRI features of central neurocytoma(CNC).Methods The clinical and MRI data of the pathologically confirmed CNC were retrospectively collected and compared,all the patients had undergone pre-surgery MRI plain and contrast enhanced scans.Patients were divided according to the location of the tumor as intra-lateral ventricle and ex-lateral ventricle group.Results Forty patients were finally enrolled,all tumors were solitary,39 of which were located in the ventricular system,including 2 7 cases located in lateral ventricle and 1 2 in the lateral ventricle,2 in third ventricle,5 in both lateral and third ventricle and 5 involving the whole ventricles;and 1 located in ex-ventricle(left temporal lobe).The tumors showed iso-intense on T1WI and T2WI with varying degrees of cystic degeneration.Hemorrhage was shown in 5 cases,calcification in 2 cases,markedly hyper-vascular enhancements were shown in 36 cases with 26 cases presented visible enhanced vessel sign.The comparison analysis between intra-and ex-lateral ventricle groups revealed that,the patients age,tumor size and Ki-67 expression were higher in ex-lateral ventricle group(P<0.05);hypervascular enhancement (P<0.05),gender ratio,cystic/cystic-solid appearance and vessel sigh(P>0.05)were lower than those in intra-lateral ventricle group.Conclusion CNC is typically located in lateral ventricle,with cystic and necrotic degeneration areas,hypervascular enhancement with visible vessels in solid portion on enhanced MR images;The ex-lateral ventricle CNC is rare,it commonly presents a larger tumor with high Ki-67 expression,and moderate enhancement with less visible vessels on enhanced MRI.
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Liver biopsy is the “gold standard” for the diagnosis of nonalcoholic fatty liver disease (NAFLD)/nonalcoholic steatohepatitis (NASH). Although liver biopsy has several limitations including invasiveness and errors in sampling and evaluation, in clinical trials on NAFLD/NASH follow-up and new drugs, liver histological evaluation is still a main and irreplaceable method for inclusion/exclusion diagnosis and assessment of primary endpoints in cohorts. It often takes 10-20 years for NASH to progress to liver cirrhosis, which is a surrogate endpoint in clinical trials for new drugs, and the NASH Clinical Research Network (NASH-CRN) system is recommended as the histological evaluation system in clinical trials for tracking NAFLD/NASH. The primary endpoint for NASH treatment is usually set as the reversal of NASH without progression of fibrosis; an alternative one is a reduction in NAS score by at least 2 points and a reduction in one or more histological parameters by at least 1 point, without progression of fibrosis, during the full-course treatment. Patients in phase 2b and 3 clinical trials should be monitored for at least 12 months, and if the improvement of fibrosis is set as the main assessment index, they should be monitored for at least 1-2 years and should be followed up for more than 6 months after drug withdrawal. Histological evaluation is affected by various factors. In order to ensure the quality of such evaluation, the length of tissue for liver biopsy should be larger than 2 cm (containing more than 10 portal areas). Staining and section preparation should be performed at the same time, and more than two experts specializing in liver pathology should perform single- or double-blinded review of liver biopsies to avoid evaluation bias.
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Liver biopsy is currently the gold standard for the diagnosis of nonalcoholic fatty liver disease (NAFLD),especially nonalcoholic steatohepatitis (NASH). Although liver biopsy has the limitations such as invasiveness,sampling error,and evaluation error,in clinical tri-als of NAFLD/ NASH and related new drugs,it is still used to evaluate the enrollment of patients into a cohort and primary and/ or secondary endpoints and cannot be replaced by other methods. The histological evaluation systems of NAFLD/ NASH mainly include Brunt system, NASH - CRN scoring system,European SAF/ FLIP scoring algorithm,and pediatric NAFLD histological scoring system,and an appropriate histological scoring system should be selected in clinical practice and research.
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Objective To examine the prevalence of chikungunya virus in brain tissue samples from rat?like animals in Xiamen, Shenzhen and Guangzhou, and to explore whether the rat?like animals are potential sources of human chikungunya fever infections and the host of the virus. Methods Rat?like animals were trapped in residential areas, city parks, hospitals, markets and schools in Xiamen, Shenzhen and Guangzhou (Yuexiu and Baiyun districts) between January 2013 and June 2016. Brain tissue samples of the trapped animals were collected under sterile. Chikungunya virus was detected by using reverse transcription polymerase chain reaction (RT?PCR). Results Totally 1092 rat?like animals were trapped, which belonged to 7 species, 3 genera, 2 families, 2 orders. Rattus norvegicus was the dominant species in the indoor environment, Rattus losea was dominant in wild environment, and 1092 brain tissue samples were collected. No detectable chikungunya virus was found in the brain tissue samples by RT?PCR. Conclusion There is a low possibility that rat?like animals act infectious sources of human chikungunya fever infections and the host of the virus.
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Objective To construct the eukaryotic expression vector of human telomerase RNA component ( hTR) and study its biological function tentatively .Methods hTR Gene was obtained by PCR from cDNA template , which was reverse transcribed from 293T mRNA and cloned into pCDNA3.0 vector.The recombinant plasmid and empty vector were trans-fected into 293T cells, and hTR expression was identified by qRT-PCR.HepG2 cells that stably transfected with pCDNA3.0-hTR were constructed and identified by qRT-PCR.These cells were used to assess the interaction of hTR with human telomerase revese transcriptase ( hTERT ) and dyskerin .Telomerase activity was also detected in HepG 2 cells transfected with pCDNA3.0-hTR.Results pCDNA3.0-hTR eukaryotic expression vector was successfully constructed by double digestion identification .The inserted fragment was confirmed by sequencing .The expression of hTR in human 293T cells and HepG2 pCDNA3.0-hTR stable cell line was identified.In addition, qRT-PCR and Western blotting results showed that hTR could interact with hTERT and dyskerin , while hTR overexpression could not regulate the telomerase activity in HepG2 cells.Conclusion The eukaryotic expression vector of pCDNA 3.0-hTR is successfully constructed and expressed.This study will contribute to the further study of cancer therapy targeting hTR .
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<p><b>OBJECTIVE</b>To investigate the correlation of serum osteoprotegerin (OPG) with the progression of nonalcoholic fatty liver disease (NAFLD) and the noninvasive prediction and diagnosis of nonalcoholic steatohepatitis (NASH).</p><p><b>METHODS</b>A total of 136 patients with NAFLD were enrolled, and their tissue samples for liver biopsy and serum samples obtained at 1 week after liver biopsy were collected; 83 healthy subjects without the symptoms of fatty liver disease proved by ultrasound examination were enrolled as controls. The physiological indicators including height, body weight, and waist circumference were measured, and body mass index was calculated. The biochemical parameters including alanine aminotransferase (ALT), aspartate aminotransferase (AST), AST/ALT, alkaline phosphatase, gamma-glutamyl transferase, total cholesterol, triglyceride (TG), high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol were measured. Double-antibody sandwich enzyme-linked immunosorbent assay was used to determine the serum level of OPG. The rank sum test, chi-square test, t-test, one-way analysis of variance, Spearman correlation analysis, least significant difference test, and receiver operating characteristic (ROC) curve were applied for statistical analysis of various data.</p><p><b>RESULTS</b>Serum OPG level was correlated with AST and TG (P < 0.05), and was highly correlated with hepatocyte fatty degeneration, ballooning degeneration, intralobular inflammation, portal inflammation, and fibrosis degree (P < 0.01). With the increasing NAFLD activity score (NAS), serum OPG level decreased, and there was a highly negative correlation between them (r = -0.928, P < 0.01). Serum OPG level was significantly lower in NASH patients than non-NASH patients. The area under the ROC curve of serum OPG level was 0.963, and according to the Youden index, its optimal sensitivity and specificity were 96.1% and 97.4%, respectively, at an optimal cut-off value of 242.96 ng/L, which suggested a high diagnostic power.</p><p><b>CONCLUSION</b>In NASH patients, serum OPG level decreases significantly. Serum OPG level can be used as an independent predictive factor to evaluate NASH and its severity, as well as a noninvasive diagnostic index for NASH.</p>
Subject(s)
Humans , Alanine Transaminase , Blood , Alkaline Phosphatase , Blood , Aspartate Aminotransferases , Blood , Biopsy , Body Mass Index , Case-Control Studies , Cholesterol , Blood , Disease Progression , Enzyme-Linked Immunosorbent Assay , Fibrosis , Inflammation , Pathology , Liver , Pathology , Non-alcoholic Fatty Liver Disease , Blood , Diagnosis , Osteoprotegerin , Blood , ROC Curve , Triglycerides , Blood , gamma-Glutamyltransferase , BloodABSTRACT
Objective To construct PES1 shRNA stable expression cell lines in tongue squamous cell carcinoma ( TSCC) cells and to study the effect of knockdown of PES 1 on the growth of TSCC cells .Methods Recombinant lentivirus carrying PES1 shRNA was packaged and obtained in 293T cells.TSCC cells (Tca8113, SCC6 and SCC15) were infected with the lentivirus and selected for stable cells .PES1 expression was identified by Western blot .The effect of inhibition of PES1 on the growth and cell cycle of TSCC cells was detected by growth curve and flow cytometry .Results TSCC cells stably expressing PES1 shRNA were constructed.Knockdown of PES1 inhibited cell proliferation and induced cell cycle ar-rest at G0/G1 phase.Knockdown of PES1 inhibited expression of cyclin D1 in TSCC cells.Conclusion Inhibition of PES1 results in reduced cell proliferation , cell cycle arrest at G 0/G1 phase and reduction of cyclin D 1 expression in TSCC cells . PES1 may be a target for TSCC gene therapy .
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AIM: To study the role of post-hemorrhagic shock mesenteric lymph (PHSML) drainage on the balance of angiotensin-converting enzyme ( ACE) and ACE2 in the kidney.METHODS:A hemorrhagic shock model was established and then fluid resuscitation was performed to the animals in shock and shock+drainage groups, and the PHMSL was drained in shock+drainage group after fluid resuscitation.After 6 h of resuscitation, the mRNA expression of ACE, ACE2, angiotensin II (Ang II) type 1 receptor (AT1R) and Mas-related G-protein-coupled receptor (MasR), and the levels of Ang II and Ang (1-7) in the renal tissues were observed.RESULTS:Hemorrhagic shock increased the levels of ACE mRNA, AT1R mRNA and Ang II, and decreased the levels of ACE2 mRNA, MasR mRNA and Ang(1-7) in the kid-ney.PHSML drainage abolished the effect of hemorrhagic shock on ACE2 and AT1R mRNA expression.Meanwhile, PHSML drainage reduced the hemorrhagic shock-induced increases in the ratios of ACE/ACE2, Ang II/Ang(1-7) and AT1R/MasR.CONCLUSION:The PHSML drainage restores the balance of ACE/ACE2, which is beneficial to alleviate acute kidney injury following hemorrhagic shock in the mice.
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<p><b>OBJECTIVE</b>To investigate the prevalence of serum antibodies against Japanese encephalitis virus (JEV) in bats.</p><p><b>METHODS</b>Blood samples from the heart were obtained from bats captured in Guangdong and Hainan Provinces in 2013. The anti-JEV antibodies in bat sera were tested using indirect ELISA and virus neutralization test.</p><p><b>RESULTS</b>A total of 201 bat serum samples were tested, in which the total positivity rate of anti-JEV antibodies was 46.27% (93/201). The positive rate of anti-JEV antibodies in bats from Hainan and Guangdong Provinces was 88.89% (48/54) and 30.61% (45/147), respectively. All the samples from Rousettus leschenaultia, Miniopterus schreibersii, Pipistrellus abramus, and Rhinolophus macrotis were positive for anti-JEV antibodies, and up to 95.56% (43/45) of the samples from Miniopterus schreibersii (from Hainan Province) yielded positive results. Of the 28 samples with positive results by indirect ELISA, 15 showed positive results in virus neutralization test (53.57%) with neutralization antibody titers ranging from 1:10 to 1:28.22.</p><p><b>CONCLUSION</b>Bats from different regions and of different species can be naturally infected with JEV and have a high prevalence of anti-JEV antibodies in their sera. The role of bats in the natural cycle of JEV awaits further study.</p>
Subject(s)
Animals , Antibodies, Viral , Blood , China , Chiroptera , Allergy and Immunology , Virology , Encephalitis Virus, Japanese , Enzyme-Linked Immunosorbent Assay , Neutralization TestsABSTRACT
Hydrogel is a creative polymeric biomaterial which can resemble extracellular matrix (ECM) in vitro. Hydrogel is also a material with intrinsic bioinert, but it can offer mechanical support and developmental guide for cell growth and new tissue organization by designing physicochemical and biological properties of hydrogels precisely. This review mainly introduces design of hydrogels, properties and applications in tissue engineering and regenerative medicine, drug delivery, stem cell culture and cell therapy.
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Humans , Biocompatible Materials , Cell Culture Techniques , Extracellular Matrix , Hydrogels , Chemistry , Regenerative Medicine , Stem Cells , Tissue EngineeringABSTRACT
[ ABSTRACT] AIM:To study the effects of soybean isoflavones on mitochondrial ultrastructure, neuronal apopto-sis and expression of cytochrome C, caspase-9 and caspase-3 in the rats with cerebral ischemia/reperfusion.METHODS:Adult healthy SD rats ( n=60 ) were randomly divided into 3 groups: sham group, ischemia/reperfusion injury ( I/R ) group and soybean isoflavone ( SI) pretreatment group.Soybean isoflavones (120 mg· kg-1 · d-1 ) were fed by gastric lav-age for 21 d.The global ischemia/reperfusion model of the rats was established by blocking 3 vessels, and then reperfused for 1 h after 1 h of ischemia.The morphological change of the cerebral cortex cells was observed under light microscope. The mitochondrial ultrastructure of the cerebral cortex cells was determined by transmission electron microscope.The apop-totic rate of the cerebral cortex cells was detected by flow cytometry.The expression of cytochrome C, caspase-9 and caspase-3 in the cerebral cortex cells was determined by semi-quantitative RT-PCR and immunohistochemical techniques. RESULTS:Disintegration of mitochondria membrane and disappearance of the mitochondrial cristae were seen in I/R group.Compared with I/R group, the change of ultrastructure of mitochondria was significantly improved by soybean isofla-vone pretreatment, and the neuronal apoptotic rate was also significantly decreased (P<0.01).The mRNA expression and protein content of cytochrome C, caspase-9 and caspase-3 in I/R group were obviously higher than those in sham group ( P<0.01).Compared with I/R group, the mRNA expression and protein content of cytochrome C, caspase-9 and caspase-3 in SI group were significantly decreased (P<0.01).CONCLUSION: Soybean isoflavones attenuate cerebral ischemia/reperfusion injury by stabilizing the structure of mitochondria, preventing cytochrome C release to the cytoplasm, inhibiting the activation of caspase-9 and caspase-3 and decreasing cell apoptosis.
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The relatively independence and separation in the teaching of clinical medicine and basic medicine limits the ability training of students,and the combination of basic and clinic medicine in teaching has become the core issues and trends in medical teaching reform.So the conventional teaching patterns were reformed in pathophysiology by applying combination of basic and clinic medicine in teaching.In order to accelerate medical students to early exposure to clinical medicine Bengbu medical college changed the teachers' teaching ideas,promoted the local integration of the courses of basic and clinical medicine and the combination of basic and clinic medicine teachers in teaching,reformed mechanisms in teaching management and teaching operation.We also tested the preliminary reform effects by way of students' evaluation of the teaching and examination,and discussed the unfavorable factors affecting the educational reform and thought out some countermeasures.
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Objective To compared two classical rat models of nephrotic syndrome and to provide some reference data to researchers.Methods Thirty male SD rats were randomly divided into control group,puromycin aminonucleoside-induced nephrotic syndrome (PAN) group and adriamycininduced nephrotic syndrome (ADR) group.The body weight,twenty four hour proteinuria level,serum albumin concentration,cholesterol concentration,creatinine and urea concentration were measured.The renal pathology change was evaluated.The drug toxic effects,administration methods and the costs were also compared.Results There was no significant difference in body weight and hair color between control group and PAN group.Compared to control group,the body weight of the rats significantly decreased at day 15 and day 21 in ADR group (P ＜ 0.01),accompanied by epilation and diarrhea.Compared to control group,the 24-hour urinary protein levels increased significantly at day 10 (P ＜ 0.01),day 15 (P ＜ 0.01),and reached the peak level at day 15 (P ＜ 0.01),day 21 (P ＜ 0.01) in PAN group and ADR group respectively.Compared to control group,the serum albumin concentration decreased significantly at day 10 (P＜0.01),and return to normal level at day 15.The serum cholesterol concentration was increased significantly at day 10 (P ＜ 0.01) and return to normal at day 15 in PAN group.Compared to control group,the serum albumin concentration was decreased significantly at day 15 (P＜0.05) and return to normal at day 21 in ADR group.No significant difference of serum creatinine and serum urea nitrogen levels were found among three groups.Compared to control group,the width of foot process increased significantly at day10 (P ＜ 0.01) and day 15 (P ＜ 0.05) in PAN group and ADR group respectively.To successfully induce a nephrotic rat model (per 100 g),the cost of PAN group was 3.1 times of ADR group (578.10 yuan vs 186.94 yuan).Conclusions Nephrotic syndrome can be induced by both PAN and ADR.The administration of PAN via intraperitoneal injection is more convenient as compared to ADR via tail intravenous injection.Compared to ADR,PAN can induce nephrotic syndrome model more rapidly,with more consistent detection index,and less toxic effects,but its cost is more expensive.
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Objective To explore the effects of the β3 adrenergic receptor (β3-AR) agonist BRL37344 on cardiac fibroblast proliferation and collagen fiber hyperplasia in Wistar rats by promoting the phosphaticlylinositol 3 kinase-protein kinase B(PI3K-Akt) signal transduction pathway. Method Cardiac fibroblasts(CFbs) were isolated from 1 - 3 day-old Wistar rats under the sterile environment in the laboratory of the First Affiliated Hospital of Hasbin Medical University, by using enzyme digestion and an modified technique of differential anchoring velocity.The cultured myocardial cells were randomly (random number) divided into five groups. ① In blank group, rats were not treated with drug; ②in BRL group, rats were treated with BRL37344; ③in LY group, rats were treated with LY294002(PI3K antagonist) for one hour before treated with BRL37344;④in Akt-Ⅰ group,rats were treated with Akt-Ⅰ (Akt antagonist) for one hour before treated with BRL37344;⑤in L-A group, rats were treated with LY294002 and Akt-Ⅰ for one hour before treated with BRL37344. MTT colorimetric method and RT-PCR were used to observe the role of β3-AR agonist with or without PI3K antagonist and/or Akt antagonist in cardiac fibroblast proliferation (CFP) and collagen fiber hyperplasia (CFH). Comparisons between groups were made by one-way ANOVA and Tukey test. Results ①β3-AR was present in the CFbs. ②Compared with BRL group, the CFP and CFH in LY and Akt-Ⅰ groups were lower (P ＜0.01) and those in L-A group were lowest (P ＜ 0.01). ③Compared with blank group,the expressions of type Ⅰ and type Ⅲ fiber mRNA obliviously increased in BRL group (P ＜ 0.01),and at 48 hours,the expressionrs reached peak. At 48 hours,compared with BRL group,the expressions in LY and Akt-Ⅰ groups were lower, and were lowest in L-A group ( P ＜0.01). Conclusions BRL37344 promotes cardiac fibroblast proliferation and expressions of type Ⅰ and Ⅲ collagen fiber mRNA by activating the PI3K-Akt signal transduction pathway.