Study On The Pathogenicity Of BALB/C Mice And The Primary Cardiac Muscle Cells Infected With A CVB3 Strain From Sichuan Golden Monkey

Backgroud:Coxsackievirus belongs to micro-RNA viraceae, the member of Enterovirus genus, since 1948 discovered, is widespread in the crowd, is one of the major pathogeny of human viral myocarditis which people pay close attention to for the past few years. CV can be separated to two subtypes, A and B. CVB has six serotypes (CVB1~CVB6). Besides human, CVB can infect orangutan and beagle, even to death. Most information report CVB3 encroachment myocardium, cause myocardium damage, especially in children. Except myocardiophilic CVB3 can cause typical myocarditis, some strain can cause pancreas, kidney, nerves, uterus damage, some strain has no pathogenicity. It can be seen that different strain can cause different pathological changes. In order to identify the pathopoiesis characteristics of CVB3 Strain from Sichuan Golden Monkey, this study by means of BALB/C experimental animal, carry out correlative study of pathogenicity .Cultivate Primary Cardiac Muscle Cells of BALB/C successfully by means of primary cell culture . Inoculate the virus in the mouse myocardial cell of primary culture, we can see the apparente pathological changes in the poison cell, including pyknosis, desquamation, lost rhythmic shrink which is myocardial cell characteristic. Collect CPE cell, centrifugate, fix in 2.5% glutaraldehyde, make virus ultramicrocut, there are a lot of crystal lattice structure caused by CVB3 nucleocapsid in electron microscope, which demonstrate the virus was massive duplicate in myocardial cell, endochylema vacuolization , most organoid lost its internal structural feature. Inoculate the virus in healthy BALB/C mouse by peritoneal injection, observe the pathogenicity to mouse. The mouse began to die after inoculate the virus 60h, death rate is 80%. Appearing typical conjunctivitis. Analyze the death mouse, we can find the most apparente pathological changes is abdominal cavity fat necrosis,liver swell, needlepoint hoariness tittle in the surface, some mouse spleen mouse,conglutination in balance and necroticfat . Select the heart, liver, spleen, lung, kidney, brain and fat pathology tissue, make paraffin section, observe its pathological change characteristic in light microscope, we can find adiponecrosis is obvious; The parenchymal organ such as heart, liver, kidney have different extent of Cell degeneration, liver neorobiosisis very severe; brain tissue has light blood clot; stomach intestine mucosa epithelium necrosis and desquamate. which indicated BALB/C Mice infected with a CVB3 Strain from Sichuan Golden Monkey can cause most organ damage. Immunity class result show, virus positive reaction in myocardial cell. make myocardial cell ultramicrocut, there is no apparente ultramicro pathological changes in electron microscope.Collect the blood of Mice infected with a CVB3 Strain from Sichuan Golden Monkey, separate the serum, detect the myocardial enzyme, liver duty, kidney duty, blood sugar, amylopsin. Result demonstrate the Erzymes in serum show different extent changing characteristic. BALB/C mouse of peritoneal injection inoculated CVB3/SGM-05, every myocardial enzyme apparentely heighten, compared to control group P<0.01, the difference extremely significant. Oral nasal way is the same as peritoneal injection, but the rangeability is lower. Except individual mouse liver duty has marked change (P<0.05), most has no significant deviation. Compared to control group, group of peritoneal injection mouse BUN significantly decrease after virus inoculation, then return to normal gradually; Oral nasal group BUN most lower thannormal; CRE higher than control group,but no significant deviation. The two groups AMY heighten significantly, P<0.01, difference extremely significant. These detection is different from CVB3infected BALB/C, suggest that clinical diagnosis of CVB3 infection, should aggregate analysis except enzymology.This study initial identify pathology characteristic of CVB3/SGM-05 to BALB/C mouse and in vitro cardiac infection,not only settle the grounding for further construct animal model of this strain,but also offer the reference for clinical hematology diagnosis of this strain infection .