Objective To compare four PCR genotyping method for Clostridium botulinum,and provide the reliable method for detection and identification of Clostridium botulinum from surveillance and foodborn poisoning in Sichuan Province.Methods Six strains of C.botulinum types A,B and E were used to compare four PCR genotyping method-one muhiplex PCR method was from US FDA,two multiplex PCR method and one real-time PCR method were from ISO,and the differences were preliminarily analyzed.Results Three multiplex PCR method could detect C.botulinum types A,B and E in a single reaction.The expected bands for type A were vague using ISO multiplex PCR method 1,whereas bright expected bands could be obtained in the identification of C.botulinum by the other two multiplex PCR method.Real-time multiplex PCR method could detect different types of C.botulinum simultaneously;however,classification should be carried out separately because fluorescent labels were the same.Conclusion Multiplex PCR method from FDA and multiplex PCR method 2 from ISO were relatively simple and could be recommended for C.botulinum surveillance in Sichuan Province.

PURPOSE: This study investigated the role of natriuretic peptide receptor 2 (NPR2) on cell proliferation and testosterone secretion in mouse Leydig cells. MATERIALS AND METHODS: Mouse testis of different postnatal stages was isolated to detect the expression C-type natriuretic peptide (CNP) and its receptor NPR2 by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Leydig cells isolated from mouse testis were cultured and treated with shNPR2 lentiviruses or CNP. And then the cyclic guanosine monophosphate production, testosterone secretion, cell proliferation, cell cycle and cell apoptosis in mouse Leydig cells were analyzed by ELISA, RT-qPCR, Cell Counting Kit-8, and flow cytometry. Moreover, the expression of NPR2, cell cycle, apoptosis proliferation and cell cycle related gene were detected by RT-qPCR and Western blot. RESULTS: Knockdown of NPR2 by RNAi resulted in S phase cell cycle arrest, cell apoptosis, and decreased testosterone secretion in mouse Leydig cells. CONCLUSIONS: Our study provides more evidences to better understand the function of CNP/NPR2 pathway in male reproduction, which may help us to treat male infertility.
Animals ; Apoptosis ; Blotting, Western ; Cell Count ; Cell Cycle ; Cell Cycle Checkpoints ; Cell Proliferation ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Germ Cells ; Guanosine Monophosphate ; Humans ; Infertility, Male ; Lentivirus ; Leydig Cells ; Male ; Mice ; Natriuretic Peptide, C-Type ; Polymerase Chain Reaction ; Receptors, Peptide ; Reproduction ; Reverse Transcription ; RNA Interference ; S Phase ; Testicular Diseases ; Testis ; Testosterone