Infection of Entamoeba histolytica with chytridiaceous fungus Sphaerlta was observed in some specimens obtained from a farmer and stained with iron-haematoxylin. The fungi were found in 78% of tne cysts, mostly immature ones. Within the amoebae this parasite occurred singly, in groups, or in the form of a sporangium. It was located in the cytoplasm, the glycogen mass or the chromatoidal bars.In the same specimen, the parasitic fungus was also found in 18% of E. coli cysts; in 11% of E. nana cysts; while only one of 16 E. hartmanni cysts was parasitized.It is an interesting case of superimposed parasitism so far reported in China as well as a rare rase of several species of amoebae being heavily involved with the same in the scientific literarure.

Purpose　The aim is to study the influence factors of recombinant E.coli cell growth,product expression and purification,in order to raise the product rate of rhGM-CSF.Methods　Three pH media were planed to observe the effects of pH on E.coli cell growth and expression with the indexes of the cell density and expression level.The influences of various treatments on product rate were observed with the indexes of the volume and purity of inclusion bodies and the yield,purity,activity of product.Using SDS-PAGE method for analysis of purity.The product bioactivity was determined by using TF1 cell and MTT colorimetric method.Results　The cell density in the medium pH 7.0 was 2.22 g/L;the product expression rate was 23%.It was two times higher than other pH media.The inclusion body prepared by bacteria without freezing store was 2.5 times volume than freezing store.During renatura-tion PEG 4000 could raise 0.8 times active product.The optimum conditions used rhGM-CSF product rate could raise 8 times.Concluston　The product rate of rhGM-CSF expressed in E.coli was affected by many factors.The optimum condition for culture,expression and purification could raise quite a number of product rate.

Objective To discuss the treatment efficacy of the femoral-head osteonecrosis by umbrella strut bone grafting and bone marrow mesenchymal stem cells. Methods The femoral-head osteonecrosis models on one side were established in forty-two goats using liquid nitrogen, which were randomly classified into three groups after 8 weeks. Group A were conducted umbrella strut bone grafting and bone marrow mesenchymal stem cells transplantation, group B were conducted simple unit side muscle bone flap grafting, and group C were conducted umbrella strut bone grafting and Cancellous bone grafting transplantation. The health side of three groups was treated as self-control. The radiological and histology examination were made at 3,6treated lateral femoral-head shape restored,cystic cnlow-density area disappeared, the original collapse had repaired, and no further collapse of the femoral-head shaped for both group A and B. Bone fusion was good and bone column shadow had been vague for group A, while the bone fusion healed somewhat less and bone column shadow still clear for group B. Group C had irregular contour of femoral head, flat collapse, uneven density, and fracture cracks could be seen at weight-bearing area of femoral head, obviously was the poorest bone had different states: mature and arranged ruled for group A, childish and irregular arrangement for group B, thinning, sparse and fracture for group C. The percentage of empty lacunae counts decreased, while the percentage of trabecular bone area fraction increased for group A. In contrast with group B, and C, the difference was statistically significant (P ＜ 0.05), but the difference was no longer statistically significant when contrast with healthy sides (P ＞ 0.05). 6 months later, the radiological and histology examination showed that:group A and group B had been no significant difference comparing to normal femoral heads, while the femoral head of group C collapsed and disappeared completely. Group A and group B were no difference comparing the healthy side (P ＞ 0.05)in the area fraction of trabecualr bone and percentage of empty lacunae counts, neither it was between group A and group B(P ＞ 0. 05). Conclusion The treatment efficacy of the femoral-head osteonecrosis by umbrella strut bone grafting and bone marrow mesenchymal stem cells was good and was better than cancellous bone grafting transplantation and simple unit side muscle bone flap grafting.

Objective To isolate and identify Cryptosporidium oocysts from feces of naturally infected cow. Methods Fecal samples were collected from Cryptosporidium infected cows confirmed by modified acid-fast staining method. Oocysts were isolated and purified with Sheather sucrose density gradient centrifugation technique. Genomic DNA was isolated with Chelex-100. Both primers were designed to amplify Cryptosporidium small subunit ribosome RNA gene (SSU rRNA) and Cryptosporidium oocyst wall protein gene (COWP), respectively. The PCR products were cloned into pGEM-T and pGEM-T Easy vector and sequenced subsequently. Homology and phylogeny were analyzed with BLASTn and MEGA software. Results The results suggested that the size of oocysts was (7.4?0.32)?m by(5.4?0.21)?m and the ratio of length and width was 1.37?0.07 (n=20). BLASTn revealed that the identity of SSU rRNA and COWP gene of Cryptosporidium isolated from cow to the counterparts of C.andersoni was 100% and 99% respectively. Phylogenetic reconstruction placed the isolated Cryptosporidium within the C.andersoni clade based on the sequence of SSU rRNA and COWP gene. Conclusion What isolated from naturally infected cow feces has been identified as C. andersoni.