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This article discusses the design and implementation strategy of Autopsy and Accident Authentication case seminar, with the aim of linking pathology to clinic medicine, theory to practice, and cultivating and improving the thinking ability of analyzing and solving problems for medical students in their medical study. The article investigates the effects and feedback of six rounds of classes, reports three aspects as strengthening the systemic understanding of basic medicine, improving the application and thinking ability, and promoting the thinking of medical humanism, and also discusses the teaching effects, students' feedback, and the significance and existing defects of the seminar.
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Objective To investigate the effect of albumin on expression of NLRP3 inflammasome and its downstream cytokines IL-1β and IL-18 in tubular epithelial cells.Methods Thirty mesangioproliferative glomerulonephritis (MsPGN) patients with different levels of proteinuria were selected, and their renal biopsy samples were stained by PAS and Masson to observe tubular epithelial cells injury and inflammatory cells infiltration.NLRP3, caspase-1, IL-1β and IL-18, as well as different inflammatory cells, were detected by immunohistostaining.In vitro, Western blotting and real-time PCR were employed to detect NLRP3, caspase-1, IL-1β and IL-18 protein and mRNA in HK-2 cells stimulated by bovine serum albumin (BSA) (20 g/L).Results In MsPGN patients with high levels of proteinuria, there were obvious renal tubular epithelial cell injury and inflammatory cells infiltration (all P ＜ 0.05), and the expressions of NLRP3, caspase-1, IL-1β and IL-18 were up-regulated compared to patients with low levels of proteinuria (all P ＜ 0.05).Furthermore, IL-1β and IL-18expressions were positively correlated with the degree of proteinuria (r=0.836, P ＜ 0.05;r=0.901, P ＜0.05).NLRP3, caspase-1, IL-1β and IL-18 protein and mRNA were significantly increased in HK-2cells stimulated by BSA compared to the control group (all P ＜ 0.05).Conclusions Albumin is able to induce NLRP3 inflammasome activation in tubular epithelial cells, which may be the mechanism of tubulointerstitial injury and inflammation caused by proteinuria.
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Abtract:Purpose To analyze epidemiological characteristics and pathological types of 1 645 renal biopsies in Jiangsu province. Methods The reports of 1 654 percutaneous renal biopsies performed from January 2009 to June 2013 were retrospectively analysed . Results 1 597 out of 1 645 renal patients were successfully biopsied with a success rate of 97. 1%. Primary glomerular diseases ac-counted for 78. 56% of the total patients, secondary glomerular diseases 18. 71%. IgA nephropathy and mesangial proliferative lession accounted for high percent of primary glomerular diseases. Lupus nephritis was the most frequent pathologic type of secondary glomeru-lar diseases, followed by allergic purpura nephritis and diabetic nephropathy. Mesangial proliferative glomerulonephritis and hyperten-sive renal injury were more common in the Southern than in the Northern Jiangsu province, while acute tubular necrosis and allergic purpura nephritis were less in the Southern Jiangsu province. Conclusions Primary glomerular disease is still the most frequent glo-merular diseases in Jiangsu province, among which the IgA nephropathy was predominated. In secondary glomerular disease, lupus ne-phritis is the most frequent pathological type. The incidences of kidney diseases have geographical variation.
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Objective To investigate the effect of advanced glycation end products(AGEs)on expression of vascular endothelial growth factor(VEGF)in rabbit retinal M?ller cells in vitro.Methods We successfully cultured rabbit retinal M?ller cells and made AGEs-BSA as well as its control in vitro.Study with M?ller cells were divided into AGEs-BSA group,AGEs-BSA control group and blank control group.AGEs-BSA group and AGEs-BSA control group were respectively treated with 5 different concentration series of AGEs-BSA and AGEs-BSA control and cultured for 3,6 and 9 days,while blank control group was incubated without any intervention.Then VEGF expression in M?ller cells was observed by immunocytochemistry(ICC).Results Compared with control group,VEGF expression in cultured retinal M?ller cells was significantly enhanced in AGEs-BSA group.And the effects were in the time-and concentration-dependent manners.Conclusions AGEs increases VEGF expression in rabbit retinal M?ller cells in vitro,which indicates that AGEs may promote the progression of diabetic retinopathy(DR)through the induction of VEGF expression.
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BACKGROUND: Astragalus root can inhibit apoptosis through reducing the release and interstitial accumulation of excitatory amino acids, alleviating calcium overloading and antioxidative effect.OBJECTIVE: Astragalus root was used to treat anoxic-ischemic brain injury in immature brain. We evaluated the effect of astragalus root on caspase-3 mRNA expression, and meanwhile, labyrinth test was employed to investigate the intervention of astragalus root on learning and memory function of mature rats after anoxic-ischemic brain injury.DESIGN: Randomized and controlled study.SETTING: Pediatric Department, Zhongda Hospital Affiliated to the Medical College of Southeast University; Pathological Department, the Basic Medical Sciences Institute of Southeast University.MATERIALS: From October 2002 to June 2003, this study was conducted at the Experiment Center of the Medical College, Southeast University.A batch of 114 seven-day-old SD rats were selected from the same brood and divided into 3 groups, namely, sham-operation group (n=18), model group (n=48) and astragalus root group (n=48). Astragalus injection was produced by Chengdu DIAO Pharmaceutical Factory, with 10 mL astragalus injection corresponding to 20 g raw material.METHODS: Animal model of anoxic-ischemic brain injury was established in model group and astragalus root group, but was not established in sham-operation group. In astragalus root group, immediately after establishing anoxic-ischemic model and at the same time point each day, 0.08 mL astragalus injection was administered intraperitoneally until the 7th postoperative day. In model group, 0.08 mL normal saline was administered at the same time points. In sham-operation group, no treatment was given. In astragalus root group and model group, animals were decollatedat 24 hours and 5 days postoperatively to take out the brains. In sham-operation group,animals were decollated and their brains were taken out at 24 hours postoperatively. In all the groups, hippocampal brain injury was detected using histopathological method combined with semi-quantified RT-PCR methods for detecting caspase-3 mRNA. Adult rats aged 90 days were used in modified y maze to examine their learning and memory functions. All these three experiments were independent.MAIN OUTCOME MEASURES:① Hippocampal brain injury in each group was evaluated using pathological method.② Caspase-3 mRNA in the ligated side of hippocampus was detected.③ Results of modified Y maze test were analyzed.RESULTS:All of the 114 rats entered the statistical analysis.① Assessment ofhippocampal brain injury in each group with pathological method:In sham-operation group, the bilateral hippocampus showed no swelling or necrosis, and neural cells in this area had normal morphological features with a density of (87.7±0.6) × 103 per high amplification field. In model group, the ligated side of hippocampus was swollen with a widened spatium and the cell density decreased to (68.8±3.0) × 103 per high amplification field, which significantly differed from that in sham-operation group (P ＜ 0.01). At the fifth day, the volume of ligated side of hippocampus reduced with pyramid layer disorganized and neural cells sparse at a density of (48.7±2.2) × 103 per high amplification field. These changes were significantly different from those of sham-operation group and the same side at 24 hours (P ＜ 0.01). At 24 hours the ligated side of hippocampus was less swollen in astragalus root group than in model group.At day 5, the whole hippocampus was observed. At these two time points,cell death rate in astragalus root group was significant lower than that in model group(P ＜ 0.01).②Caspase-3 mRNA in the ligated side of hippocampus in all the groups: In sham-operation group, the expression of caspase-3 was low, with an absorbency value of 0.220±0.009. In model group, after ischemia and anoxia its expression increased. At 6 hours, it was 11% higher than that in sham-operation group. In astragalus root group, mRNA level reached its peak, which was 260% higher than that in sham-operation group (P ＜ 0.01). The peak of mRNA continued, decreased after 48 hours and returned to baseline at 5 days and 7 days. The fluctuation of mRNA was similar between astragalus root group and model group,but the peak value at 24 hours and 48 hours in astragalus root group was 44％-46％ lower than that in model group (P ＜ 0.01). ③ Results of modified Y maze test: As compared to model group, in astragalus root group, the number of training times for meeting the standard made by the Association was significantly smaller [(45.7±2.7), (16.1±2.5) times, P ＜ 0.01] and at 24 hours after anoxia and ischemia, memory retention was significantly higher [(48.3±11.7), (80.0±9.0)%, P ＜ 0.01].CONCLUSION: Astragalus root can effectively inhibit the apoptosis of neural cells in hippocampus in immature brain after anoxia and ischemia and enhance the survival rate of them. This protective effect may be related to its inhibitory effect on the expression of caspase-3. Meanwhile, astragalus root can dramatically improve learning and memory function of the immature brain after anoxia and ischemia.
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Objective To investigate the protective role of Jidesheng Sheyao tablet in lipopolysaccharide (LPS)-induced acute lung injury(ALI) in rats. Methods Fifty male SD rats were randomly divided into 6 groups:normal control group,dexamethasone positive group,LPS model group, Sheyao groups of low, middle and high dosages successively.All rats, except normal control group, were administrated with LPS by intravenous injection to induce acute lung injury. The rats in positive group and three different dosage groups were treated by dexamethasone(3 mg?kg -1, iv), low dosage(0.8 g?kg -1), middle dosage(1.6 g?kg -1) and high dosage(3.2 g?kg -1) of Jidesheng Sheyao tablet(oral) respectively before LPS-induced ALI. Each rat in normal control group received injection of 1ml normal saline. The animals were killed after injection of LPS for 2 hours, and then the lung index was calculated, the histopathology of the lung injury was observed by light microscope, the expression of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) proteins in the lung tissues and their activity in the bronchoalveolar lavage fluid (BALF) and the lung homogenates were detected by immunohistochemistry and zymography separately. Results Compared with model group, the lung indexes were significantly decreased in the drug groups(P
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<p><b>OBJECTIVE</b>To study the effects of hypoxia and hyperoxia on the expression and activity regulation of matrix metalloproteinase-2 (MMP-2) of the hepatic stellate cell (HSC).</p><p><b>METHODS</b>The expression of MMP-2, tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and membrane type matrix metalloproteinase-1 (MT1-MMP) in cultured rat HSC under hypoxic or hyperoxic conditions were detected with immunocytochemistry (LSAB method), the contents of MMP-2, TIMP-2 in culture supernatant with ELISA and the activity of MMP-2 in supernatant with zymography.</p><p><b>RESULTS</b>(1) In the situation of hypoxia for 12 h, the expression of MMP-2 increased (hypoxia group positive indexes: 5.7 +/- 2.0; control: 3.2 +/- 1.0; P < 0.01), while TIMP-2 decreased (hypoxia group positive indexes: 2.5 +/- 0.7; control: 3.6 +/- 1.0; P < 0.05) in HSC, and the activity of MMP-2 in supernatant declined obviously (hypoxia group: 7.334 +/- 1.922; control: 17.277 +/- 7.424; P < 0.01). At the different time courses of hypoxia, the change of expression and activity of MMP-2 was most notable at 6 h. (2) In the situation of hyperoxia for 12 h, the protein contents of MMP-2, TIMP-2 in supernatant were both higher than those of the control, especially the TIMP-2 (hyperoxia group A(450): 0.050 +/- 0.014; control: 0.022 +/- 0.010; P < 0.01), and so was the activity of MMP-2 (hyperoxia group total A: 5.252 +/- 0.771; control: 4.304 +/- 1.083; P < 0.05). The expression of MT1-MMP was also increased.</p><p><b>CONCLUSIONS</b>The HSC is sensitive to the oxygen. Hypoxia accelerates the expression of MMP-2 and the effect is more marked at the early stage. Hyperoxia increases the activity of MMP-2.</p>
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Animals , Male , Rats , Cell Hypoxia , Physiology , Hyperoxia , Liver , Cell Biology , Matrix Metalloproteinase 2 , Metabolism , Rats, Sprague-Dawley , Tissue Inhibitor of Metalloproteinase-2ABSTRACT
AIM:To investigate the relevance between metallothionein(MT) and matrix metalloproteinase-2 (MMP-2) expression and activity in hepatic stellate cells (HSCs). METHODS: Fifty Kunming male mice, weighing 25 g?5 g, were randomly divided into two groups: experiment group (40 mice) and control group (10 mice). CCl4 was used to induce the hepatic fibrosis model in the experiment group. Conditioned medium of HSCs (containing MMP-2) was added at different concentrations of MT, then MMP-2 activity was detected. Liver tissue microarray was used. Collagen fibers was detected by Sirius red staining. The expressions of MMP-2 and MT proteins in liver tissues were detected by immunohistochemistry staining. Gel zymography was used to confirm the activity of MMP-2 in liver tissue homogenate and in the conditioned medium. RESULTS: The expressions of both MMP-2 and MT proteins in liver tissues increased fluctuating with the process of fibrogenesis in the model mice alternately, but the proteins showed out of phase changes. The activity of MMP-2 in the liver tissues also increased gradually and fluctuated with the development of fibrosis. Apart from individual time points, the activity of MMP-2 and MT expression were negatively correlated. The activity of MMP-2 in the conditioned medium treated by MT declined in a dose-dependent manner (r=-0.9990, P
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AIM: To explore the expression of caspase -3 (cysteinyl aspartate-specific proteinase) in the neonatal rat cerebral cortex a nd hippocampal after hypoxia-ischemia. METHODS: Sham and hypoxia-ischemia (HI) groups were set up. The neonatal HI procedure was performed in 7-day-old rat pups. The double-lateral co rtex and hippocampal was subjected to pathological assessment, immunohistochemic al staining with caspase-3 antibody and half-quantitative reverse transcription and polymerization chain reaction (RT-PCR) to measure the change in caspase-3 pr otein and mRNA expression. RESULTS: Caspase-3 mRNA in ipsilateral cerebral cortex and hippo campal increased immediately after HI followed by a partial recovery. Thereafter caspase-3 mRNA and protein simultaneously increased with a maximum reached at 2 4-48 h after HI. CONCLUSION: Caspase-3 may play a key role in the development of apoptotic hypoxic-ischemic brain damage (HIBD) in immature rats. Neuroprotective medicine should be used before 24-48 h after HI.