ABSTRACT
The objective of present study was to develop a simple and reliable voiding spot assay (VSA) system to evaluate the lower urinary tract function of mice, and to establish it as a standardized protocol. Ultraviolet (UV) light was used to screen out the filter paper without autofluorescence and with optimal urine diffusion properties. Next, the appropriate wavelength of UV was determined based on the quality of the photographic image of urine spots on the filter paper. To confirm that the urine stain area on the filter paper was correlated with the amount of urine, a volume-area standard curve was constructed. The utility of this VSA system was validated using female wild-type C57BL/6J mice aged 12-13 weeks, and the data generated under identical procedural settings were compared among laboratories. Furthermore, this VSA system was employed to analyze the changes in voiding patterns in mice with urinary tract infections or transportation stress. No. 4 filter paper with a thickness of 0.7 mm was identified as the most suitable material for VSA, exhibiting no autofluorescence and facilitating optimal urine diffusion. The filter paper retained its integrity during the assay, and there was a linear correlation between urine volume and stained area under 365 nm UV light. Utilizing this VSA system, we determined that female wild-type C57BL/6J mice produced approximately 695.8 μL total urine and 5.5 primary voiding spots (PVS) with an average size of 126.4 μL/spot within 4-h period. Over 84% of PVS volumes ranged from 20 to 200 μL. Notably, PVS volumes of mice were similar across different laboratories. Mice with urinary tract infections or transportation stress exhibited significant changes in VSA parameters, including increased voiding frequency, PVS number, and decreased PVS volume. Therefore, this VSA system can be used to evaluate the urinary function of normal mice, as well as those with urinary tract infection or transportation stress.
Subject(s)
Mice , Female , Animals , Urodynamics , Mice, Inbred C57BL , Urination , Urinary Bladder , Urinary Tract InfectionsABSTRACT
Objective:To observe the effects of Huazhuo Jiedu Shugan Prescription （HZJDSG） on learning， memory， and the expression of phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt）/glycogen synthase kinase-3<italic>β</italic> （GSK-3<italic>β</italic>） pathway-related proteins in epileptic rats， and to explore its possible mechanism. Method:Forty-eight SPF male SD rats were randomly divided into a normal group， a model group， a sodium valproate （0.19 g·kg<sup>-1</sup>） group， and low- （2.7 g·kg<sup>-1</sup>）， medium- （5.4 g·kg<sup>-1</sup>）， and high-dose （10.8 g·kg<sup>-1</sup>） HZJDSG groups， with eight rats in each group. The normal group received 0.9% sodium chloride solution （0.035 g·kg<sup>-1</sup>） by intraperitoneal injection， and the other five groups received pentetrazol （PTZ） at the same dose to induce a chronic epilepsy model for a total of 14 times. The drug groups received corresponding drugs and the normal group and the model group received 0.9% sodium chloride solution at the same volume once a day for 28 days. During the drug intervention period， epilepsy was maintained in each modeling group by intraperitoneal injection of PTZ on day 7， 14， 21， and 28. The behavioral changes of rats were observed by Morris water maze and the pathomorphological changes of rat hippocampal neurons by hematoxylin-eosin （HE） staining. The protein expression of phosphorylation Akt（p-Akt）and p-GSK-3<italic>β</italic> was detected by immunohistochemistry and the protein expression of PI3K， Akt， p-Akt， GSK-3<italic>β</italic>， and p-GSK-3<italic>β</italic> by Western blot. Result:Compared with the normal group， the model group showed prolonged platform finding time （<italic>P</italic><0.01）， reduced number of platform crossings （<italic>P</italic><0.01）， structural damage of neurons in the CA1 region of the hippocampus， down-regulated protein expression of p-Akt and p-GSK-3<italic>β </italic>in the CA1 region of the hippocampus （<italic>P</italic><0.05）， and reduced relative expression of PI3K， p-Akt， and p-GSK-3<italic>β</italic> in the hippocampus （<italic>P</italic><0.01）. Compared with the model group， the sodium valproate group and the HZJDSG groups showed shortened platform finding time （<italic>P</italic><0.01） and improved neuronal structure in the CA1 region of the hippocampus， while the sodium valproate group and the high- and medium-dose HZJDSG groups exhibited increased number of platform crossings （<italic>P</italic><0.01）， up-regulated protein expression of p-Akt and p-GSK-3<italic>β</italic> in the CA1 region of the hippocampus （<italic>P</italic><0.05）， and elevated relative expression of PI3K， p-Akt， and p-GSK-3<italic>β</italic> （<italic>P</italic><0.01）. Conclusion:HZJDSG can improve the learning and memory of epileptic rats， and its antiepileptic effect may be achieved by the activation of PI3K/Akt/GSK-3<italic>β</italic> pathway-related proteins.
ABSTRACT
OBJECTIVE@#To analyze the causes of positive irregular antibody screening test and incompatibility of cross matching in one patient with autoimmune hemolytic anemia complicated with neonatal hemolytic disease, and to accurately identify the type of antibodies in patients, and to select a reasonable strategy for blood transfusion.@*METHODS@#One children was enrolled, blood group positive and reverse typing, Rh typing, direct anti-human globulin test, free test, dispersal test and cross matching test were carried out by test tube method and microcolumn gel card; irregular antibodies were identified by the reaction of DTT treatment and untreated panel cells with patients' plasma.@*RESULTS@#The blood group of the patient was RhD positive B and irregular antibody screening positive, while the blood group of the mother was RhD positive O and irregular anti-screening negative, the result showed that the anti-LW detected in the plasma of the patient was autoantibody and ABO neonatal hemolytic disease (ABO-HDN) was present. Both O type RhD positive washing RBCs and B type RhD negative RBCs were transfused effectively.@*CONCLUSION@#Irregular antibodies in patients are anti-LW antibodies, and transfusion of homotype RhD negative suspended erythrocytes after the exclusion of ABO-HDN shows a better effect.