| Objective1.To detect AKP and Calcified material in eggs deposited in liver and colon of mice with Sj infection before and after routine treatment of PQZ, in the hope of being markers for them to discriminate dead or live eggs and providing research basis on clinical therapeutic effect assessment.2.To observe pathology damage of liver and colon of different stages through examination of PDI and ATPase in liver and colon of mice with Sj infection before and after treatment,so as to add an effective index for evaluating schistosomiasis prognosis.Methods1.63healthy adult KM mice were rondomly divided into seven groups, namely normal group,infection groups(three times of45d,90d,120d) and treatment groups(three times of1M,2M,3M,).At the end of the model,liver and colorectal mucosa of mice were taken to made whole eggs,frozen sections and paraffin sections.2.Do AKP dyeing with the azo coupling method and NBT/BCIP and VON KOSSA,conunt the number of positive eggs and calculate the positive rate to analyse the effect of discriminating eggs of the two mathods with SPSS.3.observe and compare pathomorphological changes of tissues with routine HE and MAS SON dyeing, and the expression levels of PDI and ATPase with immnohistochemistry method.Results1.The Sj worms in each group:male-female pairing worms of different numbers were found in all infection groups,while in the treatment groups no live worms were found.2.the positive rate of eggs in145d group was above95%, and decreased gradually with the infection prolonged.the positive rates in T2M and T3M ramatically reduced,close to zero.the two AKP mathods represented no statistical difference between examination effect(p>0.05).the positive rate of eggs showed positive correlation between liver and colorectal mucosa,as the spearman rank correlation coefficient was0.876(p<0.01).3.calcification content tests of eggs in paraffin sections of liver and intestine tissue showed that,in the I45d group no eggs were positively colored,calcification rate of eggs in I90d and I120d groups were48%and45%(intestine) and51%and62%(liver), respectively.In TIM, T2M and T3M groups, the worm eggs calcification rates were45%,95%and95%(intestine) and22%,86%and96%(liver).calcification rate of eggs in each group between liver and intestine was positively related(correlation coefficient is0.841, p<0.05), but very few eggs in T3M group still presented calcified.4.There were only a few collagen deposition in liver and intestine of I45d group,along with infection time prolonged, the collage scale grew, distributed mainly surrounding eggs and in liver plate. While obvious collagen still existed in PQZ treatment groups.5.there were statistical differences among infection groups, treatment groups and normal group in liver of PDI expression. The expression of PDI was highest in I90d(p<0.05compared with each group) and reduced in treatment groups, each group had an enhanced expression of PDI in colorectal mucosa,compared with N. levels in I90d and I120d of PDI significantly higher than the others(p<0.05),and in treatment group the levels were decreased notably.6.ATPase activities increased in infection group,mainly expressing in or around the egg granuloma nodules.there were a statistical difference in infection groups,T1M and the others.No difference were found in the three infection groups,also were in T2M,T3M and N (p>0.05).ConclusionVisible fibrosis still developed in liver and intestine tissue of mice treated with PZQ effectively after infection45days, which suggest that we have to focus to both anti-pathogen and anti-fibrosis to therapy schistosomiasis. Detection of AKP and calcification of eggs had a similar efficiency to discriminate dead or live eggs. Expression levels of PDI and ATPase increased along with Sj infection period prolonged, and decreased along with treatment period extension, preliminarily showed that the two molecules may not only involved in pathological process, but also have an important effect in evaluating prognosis or outcome of schistosomiasis. |
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