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· AIM:To compare the effect of fixation and select the optimal fixation solution and condition for PAS staining of guinea pig eyes among three different fixation solution:4％ paraformaldehyde solution,4％ glutaraldehyde solution and Davidson solution.· METHODS:Totally 30 healthy guinea pigs were divided into 6 groups:Group Ⅰ-Ⅱ were fixed in 4％ paraformaldehyde solution and 4％ glutaraldehyde solution for 24h,respectively;Group Ⅲ-Ⅴ were fixed in Davidson solution for 3,6 and 24h,respectively;and Group Ⅵ were fixed in Davidson solution for 3h and then transferred into 10％ neutral formaldehyde solution for 48h.All groups were sectioned by routine section method and undergone PAS staining,and then observed by light microscope.· RESULTS:It was found that the group which was fixed by Davidson solution for 3h,remained the most complete structure for PAS staining (Group Ⅲ).While the effect of fixation for the group which was transferred into 10％ neutral formaldehyde solution for preserving for 48h after fixing in Davidson solution for 3h was also acceptable for PAS staining (Group Ⅵ).The retinal cells remained clear and in order for both groups which was mentioned above.· CONCLUSION:The best fixation condition for PAS staining for eyes of guinea pigs is fixation in Davidson solution for 3h among these fixation conditions,while it is also suitable to transfer the eyes into neutral formaldehyde solution after fixing in Davidson solution for 3h for preserving for long periods,which is not severely reduced the effect of fixation for PAS staining.
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Objective To investigate the diagnosis and treatment of pulmonary alveolar proteinosis (PAP). Methods Clinical data of 3 children with PAP admitted from 2010 to 2016 were retrospectively analyzed. Results Three children were female and aged 3 years and 1 month, 3 years and 9 months, and 6 years and 4 months, respectively. The main symptom of the three children was cough along with or without anhelation. Two of them had longer course of disease. The therapeutic effect of conventional treatment was poor, and their chest CT indicated significant lesions. Fiberoptic bronchoscopy showed positive reaction by PAS staining, and the treatment effect was improved after bronchoalveolar lavage. Conclusions The main clinical symptoms of PAP are non-specific cough and shortness of breath. PAS staining and bronchoalveolar lavage are of great value in the diagnosis and treatment of PAP.
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Objective:To observe the bone marrow mesenchymal stem cell isolation,identification methods and protection mechanisms in the kidney of diabetic rats.Methods: Thirty-one rats were used to isolate bone marrow mesenchymal stem cells (BMSCs) by adherent culture method.Cell morphology was observed by phase contrast microscope.The surface molecular weight of cultured cells was detected by Real-time PCR.30 rats were induced by intraperitoneal injection of streptozotocin to establish diabetic rat model (n=15).And stem cell treatment group (n=15) according to the different treatment methods.Rats in the control group were treated with insulin combined with probucol.Stem cell treatment group was implanted with stem cells.Before and after treatment,fasting blood glucose,24 h urinary protein excretion,creatinine clearance rate and kidney weight were measured.PAS staining.And the renal tissue was detected by Western blot.Results: The primary bone marrow mesenchymal stem cells (MSCs) were cultured for 24 hours,and the cells were irregularly distributed in the culture flask.After 7 days of culture,the cells were regular,elongated,elliptical,and strong refraction,and the cells were fused with each other.The morphology of BMSCs was uniform and showed a long fusiform shape,CD44,CD31 and CD34 were observed by RT-PCR.The levels of fasting blood glucose,24 h urine protein excretion and creatinine clearance rate in the observation group were significantly higher than those in the control group (P0.05).Conclusion: It is suggested that BMSCs can be isolated from rat bone marrow mesenchymal stem cells by using adherent culture method and labeled with green fluorescent protein in vitro.Bone marrow mesenchymal stem cells can protect the kidney and inhibit the oxidative stress of diabetic kidney.
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We have developed a method for estimating the depth and intensity of muscular unit represented as equivalent current dipoles by the inverse analysis of surface electromyograms (EMGs) . In this study, the validity of the locations of current dipoles estimated through the inverse analysis was verified by animal experiments. Surface motor unit action potentials (MUAPs) were recorded from the gastrocnemius muscle activated by electrical stimulation at the ventral root of lumbar spinal cord (L4 or L5) of rats. After recording the surface MUAPs for the inverse analysis and glycogen depletion of active muscle fibers by repeated electrical stimulation, periodic acid-Schiff (PAS) staining was used to determine the position of muscle fibers belonging to an active single motor unit. In the results of the inverse analysis, the values of ‘goodness of fit’ between measured and calculated MUAP were 71%, 79% and 85%. Estimated depths of current dipoles ranged from 1.8 mm to 5.9 mm. The locations estimated through the inverse analysis were more medial and shallower than the actual distribution of active muscle fibers determined by PAS staining. These errors were probably caused by the effects of the boundary in the model, the relationship between the measurement area and the location of an active motor unit, and the artifacts such as deformation of the muscle during dissection and freezing.
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Objective:To compare Lyon′s PAS standardized staining and traditional PAS staining.Methods:The sections of the same tissue were stained with the above two methods simultaneously.The results and replicability of the two methods were observed.Results:The posititive products and replicability of Lyon′s PAS staining were stronger than that of traditional PAS staining in the same tissue-sections.Coclusion:Lyon′s PAS standardized staining method is better than the traditional PAS staining method.