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Objective To investigate the protective effect and mechanism of active vitamin D3 on podocyte injury in type 1 diabetic rats.Methods Animals were randomly divided into normal control group (NC group),diabetic nephropathy group (DN group),diabetes nephropathy plus active vitamin D3 group (DN + VD group).Random tail vein blood glucose was measured and 24 hours of urine was collected every 3 weeks to observe the dynamic changes of blood glucose and 24-hour urine volume and urinary albumin.Rats were sacrificed at the end of 18th week,the kidney weight to body weight ratio,serum creatinine,blood urea nitrogen,serum calcium,and serum phosphorus levels were measured.Pathological in glomeruli were observed by PAS staining.Immunohistochemistry and Western blotting were used to observe the expression of slit diaphragms proteins including Nephrin,Podocin,and vitamin D receptor protein VDR.The mRNA level of autophagy-related protein P62 was detected by realtime quantitative PCR,and expression of autophagy-related protein including LC3B/A,Beclin1,and P62 were detected by Western blotting.Ultrastructure of podocytes and autopbagosomes in podocytes were observed by electron microscopy.Results Levels of serum creatinine,blood urea nitrogen,and blood glucose in diabetic rats were higher than those in NC group (P＜0.05),but without significant difference between DN and DN+VD groups (P＞0.05).Compared with the DN group,the urinary protein and kidney weight to body weight ratio in the DN +VD group were significantly lower (P＜ 0.05).Mesangial matrix hyperplasia and basement membrane thickening were improved,and podocyte fusion and shedding were partially reversed.The expressions of Nephrin,Podocin,VDR,LC3B/A and Beclin1 were increased,and P62 mRNA and protein were down-regulated (P ＜ 0.05).The number of autophagosomes in podocytes increased.Besides,positive correlations were found between Nephrin and Beclin 1 (r =0.939 8,P＜0.05),as well as Nephrin and VDR (r=0.948 3,P＜0.05),and Beclin1 andVDR (r=0.9093,P＜0.05).Conclusion Active vitamin D3 inhibits the injury of diabetic nephropathy podocytes by up-regulating VDR expression and enhancing autophagy activity,thereby reducing proteinuria and delaying the development of diabetic nephropathy.
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OBJECTIVE@#To investigate the mechanism.@*METHOD@#We isolate the mouse bone marrow cells and cultured with rGM-CSF and rIL-4 to stimulate bone marrow cells to transfer to immature dendritic cells. And then the immature dendritic cells were costimulated with ILPS and different concentrations of Bimingan Granule.@*RESULT@#MHC II, CD80, CD86 were detected by flow cytometer and TNF-alpha, IL-1beta, IL-6 and IL-10 were quantified by ELISA.@*CONCLUSION@#Our results showed that Bimingan Granule may significantly inhibit the differentiation of immature dendritic cells to mature dendritic cells.
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Animals , Mice , Cell Differentiation , Cells, Cultured , Dendritic Cells , Allergy and Immunology , Drugs, Chinese Herbal , Pharmacology , Mice, Inbred C57BL , Rhinitis, Allergic , Drug Therapy , Allergy and ImmunologyABSTRACT
Objective:To investigate the curative effect of electroacupuncture on urine retention following cervical cancer operation. Methods: A treatment group was treated with electroacupuncture according to classification based on syndrome differentiation and a control group, by a conventional method. One course of treatment consisted of 5 days. Results: The curative rate and cure rate were 96.67% and 76.67%, respectively, in the treatment group and 70% and 36.67%, respectively, in the control group. There were significant differences in curative rate and cure rate between the two groups (P＜0.01). Conclusion:Electroacupuncture is a good method for treating urine retention subsequent to cervical cancer operation.
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BACKGROUND:Crainocerebral missile wound (CMW) is the common severe trauma of war. Environmental factor is the important factor that aggravates the development of sickness and threatens the life of the sick and wounded at early stage. The study on basic theory and treatment of CMW under hyperthermia and high-humidity is the major task in military medicine.OBJECTIVE: To observe the characters of bacterial growth in wound cavity and peripheral tissue after CMW in cats under hyperthermia and high-humidity so as to provide the reference data to the tissue repair and its functional recovery in CMW.DESIGN: Randomized controlled experiment.SETTING: Surgery Department of Neurology in Zhujiang Hospital affiliated to Southern Medical University(First Military Medical University).MATERIALS: The experiment was performed in Department of Tropical Medicine and Hygiene in Southern Medical University from April to May 2003. In the experiment, 24 hybrid cats were employed, of clean grade, of either sex, body-weighted (2.5±0.2) kg, provided from Experimental Animal Center of Southern Medical University. The animals were randomized into 4 groups, named the control at common temperature (5 cats), common temperature group (8 cats), hyperthermia and high-humidity group (8 cats) and blank control (3 cats).METHODS: After weighted, the cat was injected abdominally with 30 g/L pentobarbitol sodium solution at 30 mg/kg. After successful establishment of craniocerebral blindgut wound model, in common temperature group, the cats were placed in simulated climatic cabin at 25 ℃ and 50% relevant humidity for 6 hours and in hyperthermia and high-humidity group, the cats were placed in the hyperthermia and high-humidity environment for 6 hours. With biopsy, the fragmented brain tissue in wound tract and the brain tissue on the wall of wound tract far from 0-5 mm and 5-10 mm were collected and preparedinto successively natural solution and homogenate, and counting was done in 24 hours of bacterial culture to calculate bacterial content in wound cavity and peripheral tissue. In blank control, the cats were sacrificed with anesthesia and the material was collected directly, the rest steps were same as the control at common temperature.MAIN OUTCOME MEASURES: Bacterial content in wound cavity and peripheral tissue in each group.RESULTS: Totally 23 animals were employed in outcome analysis. Bacterial contents in same regions were different remarkably in different groups (F=171.423, P=0.000). It was indicated in paired comparison that significant difference happens between the control at common temperature, common temperature group and hyperthermia and high-humidity group and blank control successively (P ＜ 0.01). The difference was not significant in the comparison among the control at common temperature, common temperature group and hyperthermia and high-humidity group (P ＞ 0.05). Bacterial contents at different distances from the wound cavity were different remarkably in each group (F=14.865, P = 0.000).CONCLUSION: Hyperthermia and high-humidity does not impact significantly the bacterial growth of CMW in 6 hours and bacterial reproduction has not entered high-rate increasing stage yet, due to which, it is still safe for debridement repair.
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Free lysine in a preparation of queen bee's embryo was isolated using cation exchange resin- The Q-aminoand carboxyl group of lysine was complexed with cupric ion and the E-amino group reacted with FDNB(Fluoro-dinitrobenzene). After acid treatment and extraction with ether,the absorbance of the solution at 395nm was determined by spectrophotometry. Variation coefficient of the determination was less than 6% and the recovery of lysine was 95 % ~ 110 %.
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AIM To investigate the positive inotropic effect of aconitine on isolated hearts from cardiac dysfunctional rats when combined with pinacidil. METHODS In our experiment Langendorff perfusion equipment was used to investigate the following cardiac indexes:LVSP(left ventricular systolic pressure), LVDP(left ventricular diastolic pressure), +d p /d t max (the maximum going up rate of left ventricular pressure) and -d p /d t max (the maximum going down rate of left ventricular pressure) from rat hearts under two conditions:① aconitine only,② aconitine plus pinacidil(an agonist of K ATP channel) to observe the positive inotropic effect of aconitine on cardiac dysfunctional rat hearts. RESULS ①Aconitine had certain positive inotropic effect on isolated hearts from cardiac dysfunctional rats; ②Combing aconitine with potassium channel activator markedly winded the doses range between the effective and toxic concentration and enhanced the cardiotonic effect of aconitine. CONCLUSION Aconitine had positive inotropic effect on dysfunctional isolated rat hearts; the combination of aconitine with potassium channel activator markedly widened the cardiotonic doses range of aconitine and enhanced its cardiotonic effect.
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AIM: To investigate the role of mitochondrial calcium uniporter (MCU) in the cardioprotection by hypoxic preconditioning (HPC) and its relationship to mitochondrial permeability transition pore (MPTP). METHODS: Intraventricular balloon technique was employed to measure the left ventricular developed pressure (LVDP), the maximum rise/fall rate of left ventricular pressure (?dp/dt_ max ), and the left ventricular end-diastolic pressure (LVEDP) in Langendorff isolated rat heart. The hypoxia was achieved by ligation of left anterior coronary artery for 30 min followed by release of ligation for 120 min as reoxygenation. Hypoxic preconditioning was set as two episodes of 5 min global hypoxia and 5 min reoxygenation. RESULTS: Both HPC and treatment with ruthenium red (5 ?mol/L) during the first 10 min reoxygenation improved recovery of LVDP, ?dp/dt_ max and decreased LVEDP, which was associated with reduced infarct size and lactate dyhydrogenase release. These protective effects were attenuated by treatment with spermine (20 ?mol/L) during the first 10 min reoxygenation. Administration of cyclosporin A (0.2 ?mol/L) during the last 5 min of hypoxia period and first 15 min of reoxygenation period reduced the injury effect by spermine. CONCLUSION: These results indicate that inhibition of MCU is involved in the cardioprotection of HPC via inhibiting MPTP.
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AIM: To investigate whether tumor necrosis factor ? (TNF?) pretreatment can inhibit mitochondrial permeability transition pore opening in isolated rat hearts subjected to hypoxia and reoxygenation. METHODS: Isolated perfused rat hearts were subjected to 30 min regional hypoxia (occlusion of left anterior descending artery) and 120 min reoxygenation. The infarct size, lactate dehydrogenase (LDH) release during reoxygenation and ventricular hemodynamic parameters were measured. RESULTS: Pretreatment with TNF? at concentration of 1?104 U/L for 7 min followed by 10 min washout reduced the infarct size and LDH release, and improved the left ventricular performance (left ventricular developed pressure and rate-pressure product) and left ventricular end-diastolic pressure during hypoxia and reoxygenation. Administration of atractyloside (Atr, an opener of mitochondrial permeability transition pore, 20 ?mol/L) for 20 min (last 5 min of hypoxia and first 15 min of reoxygenation) and paxilline (Pax, a calcium activated potassium channel antagonist, 1 ?mol/L) for 5 min before hypoxia attenuated the reduction of infarct size and LDH release and improved the left ventricular performance induced by TNF?. CONCLUSION: The findings indicate that in the isolated rat heart model, TNF? protects myocardium against hypoxia and reoxygenation injury via inhibiting mitochondrial permeability transition pore opening as well as activating calcium, activated potassium channel.
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The spleen cells from the BALB/C mouse immunized with the type B toxoid were fused with aNS-1 myeloma cell line. The superatant containing the hybridoma-formed cells with growing pores wasscreened by EL1SA, in which 66.7% the pores could secrete the specific antibodies against botulin.After a Subclonizing culture by the limiting dilution technique four hybridoma cell lines(3B10, 3B11, 3G12 and 4A5) were established and could secrete the specific antibodies persistently in the culture medium. in which antibody titers came to 10-3-10-5, while they were injected into the BALB/C mice intrape ritoneally ascites rich in antibodies with a titer of 10-5-10-8 was produced. The results testing the (our monoclonal antibodies with the type A and B toxoids showed that the antibodies of 3G12and 4A5 were specific for the type B toxoid, and those of 3B10 and 3B11 had light cross reaction with the type A toxoid. Identification of 1g showed that the antibodies of 3B10 and 3G12 were of IgG1, while those of 3B11 and 4A5 of IgG2. The chromosomal assay confirmed the four cell lines to be hybridoma. The neutroligation test in mice revealed that those four monoclonal antibodies did not show any protective effects on botulin.