The Existence Of The ICR Mice Kidney Circadian Rhythm And Existence Significance

Objective:In this experiment, intervention was made to the black boxed mice forthe purpose of observing the expression of circadian genes: Bmal1、Clock、Cry1-2、Per1-2and Clock controlled genes DBP、Rev-erbα、Rev-erbβin themouse kidney, in order to verify the existence of circadian rhythm on themouse kidney.Method：1、Make sure the black box contains adequate ventilation and alsois strictly shaded with dark cloth.2、Select24mice that were born after3months. Weight them—theweight should not exceed3g. Put them inside the black box and providethem with enough food and water. After72hours, randomly take out twomice at a time for every4hours and kill them by breaking their necks.Then take out their kidney tissues and store them in the liquid nitrogen.3、Extract and quantify mRNA. Utilize3ug for reverse transcriptionand dilute the product concentration to300ul.Use regular PCR to test theexpression of target genes. Then utilize RT-PCR to detect the expressionof clock genes such as Clock、Bmal1、Cry1、Cry2、Per1、Per2and Dbp.4、Collect and organized the data from above experiment. Use GraphPadstatistical software for data analysis and graphics. Result：Utilize β-action as a reference gene, we observed the ups and downsof the expression of Bmal1, Cry1，Cry2, DBP, Rev-erb α, Rev-erb β, Per1over time in a rhythmic fashion, while Clock, Per2did not show any clearsign of circadian rhythm.Conclusion：Circadian gene and clock control existed in the kidneys of ICR micebut also it is expressed in a rhythmic pattern changing with time.The expression of circadian rhythm fromthe proteinuria of patientsObjective:In this experiment, we observe three different kidney disease clinicalpatients urinary protein excretion changes, discuss the differentpathological types of kidney disease patient albuminuria excretioncondition and its relations with variable degrees of kidney disease.Method：1、Based on the clinical charts of patients, select all subjectswith normal creatinine level and urine protein concentration greater than3.5g in24hr. Select3patients who did renal biopsy that shows only mildpathologic changes,3with renal insufficiency syndrome and3with uremia.2、Collect urine sample from above patients at7-8:00am and4-5:00pm.Take a certain amount of each urine sample and test its creatinineconcentration with creatinine measurement kit. Use the same amount for running the urine protein electrophoresis, then stain and discolor thegels to reveal changes in the urinary proteins.3、Collect and organize the data obtained. Utilize SPSS17.0statistical software for data analysis.Result：In patients with mild renal disease, the concentration of proteinsin the proteinuria in the morning is lower than the concentration of thatin the afternoon. In patients with renal insufficiency syndrome, thereis no difference between the morning and the afternoon urine proteinconcentrations. In patients with uremia, the urine protein concentrationis higher in the morning than in the afternoon.Conclusion：There is rhythm existed in the urine proteins. Renal pathology typesfor minor lesions in patients with urinary protein excretion in themorning is lower than the afternoon,as the patient’s condition worse,when to uremia, the trend becomes the opposite to the mild disease patients,mean the morning urine protein excretion is higher than in the afternoon.