The effects of nitroquine on the sporogonic development of Plasmo-dium yoelii were observed under electron microscopy. The female mosquitoes were fed directly with 10% sucrose solution containing 0.1%Nitroquine.It was found that the oocysts were smaller and markedly degenerated as compared with that of the control. The surface of the oocysts was rough and uneven. Under a transmission electron microscope, the cytoplasm of the affected oocysts contained vacuoles; the membane of mitochondria and uncleus was damaged; and the number of residual bodies increased.No sporoblast formation was seen in most of the affectes oocysts. The nuclear membrane of the degenerated sporozoites was thickened and the density of nuclear matrix decreased markedly as compared with that of the control. These results indicate that the nucleus and the membrane are mainly affected during the sporogonic development of P. yoelii by nitroquine.

Anopheles Stephensi,infected with Plasmodium yoelii.was fed on 10% sucrose solution containing 1%.difluoromethylornothine(DFMO)to induce the degeneration of its oocysts.On the 11th day after infection,the effects of hemocytes on the normal and degenerated oocysts were observed wtih transmission electron microscopy.In the control group.no hemacytes could be found around the normally-developed oocysts except those degenerated ones.In the DFMO group,all the oocysts underwent degeneration in various degrees and some of them were melanized.All the oocysts were attached by one or more hemocytes which belonged to the category of granulo-cytes morphologically.There were many granules with microtubular construction in the cytoplasm of the hemocytes and in the spaces between a hemocyte and an oocyst.The findings indicate that the degeneration of oocysts can exert a taxic effect on hemocytes and the latter may release the granules and possibly other substances to result in the encapsulation and melanization of the oocysts.

Objective To clone, locate and differentially analyze the transcription factor Relish gene from Anopheles stephensi, and to examine its signals-modulating action on prophenoloxidase cascade and melanization of Plasmodium yoelii oocysts. Methods Relish cDNA of total mosquitoes was amplified by RT-PCR with degenerated primers. Target PCR product was purified,cloned,sequenced and identified. Special Relish gene was amplified with specific primers from hemocytes or midgut,respectively. Semi-quantitative analysis was made under different feeding conditions. Relish message ribonucleic acid was identified with hybridization in situ. Results One cDNA segment of Relish similar to An.gambiae was acquired from An. stephensi. The same Relish gene was also manifested in the hemocytes and midgut. Marked up-regulation expression of Relish was observed at 6,12,24 or 48 h of Plasmodium yoelii infection and at 12 and 24 h after sucking nitroquine-acetate sucrose solution,that was before inducible oocyst melanization. Relish was also expressed in the hemocytes and midguts by ISH. Conclusion Transcription factor Relish of An. stephensi might play a role in signal modulation of Plasmodium yoelii infection and oocyst melanization.

The percentage of sporozoites developed in exoerythrocytic forms in the rats treated intravenously with cyclophosphamide 24 hrs before inoculation of sporozoites was higher than that in untreated controls, so was the mean size of exoerythrocytic form. Cyclophosp-hamide could decrease the sensitivity of hepatocytes to the vector's tissues inoculated together with sporozoites, thus decreasing the incidence of cloudy swelling of hepatocytes induced by the vector's tissues and inhibiting the white blood cells infiltration induced by the ruptured mature exoerythrocytic form.The fact that cyclophosphamide might enhance the invasion of sporozoites into the hepatocytes indicates that the phagocytic activity of Kupffer's cells to destroy the sporozoites might be inhibited by cyclophosphamide.

Objective To prepare and optimize formulation of ATRA liposomes. Methods ATRA liposomes was prepared by reverse evaporating method and to determine the concentration and the entrapment efficiency of ATRA liposomes by HPLC. Results The entrapment efficiency of the three ATRA liposomes formulations reached 33.98%, 42.11%, 70.34% respectively. The liposome was stable. The ATRA content of three formulations almost showed no change during the observing days. Conclusion The technique of preparing the ATRA liposomes is feasible and the method of quality control is simple and accurate.

Objective To investigate the association of plasma metallothionein 3 (MT3) and its polymorphisms with childhood autism,in order to provide the objective evidence for autistic etiology and molecular diagnosis.Methods A total of 132 autistic children were recruited from several special autism training schools in Wuhan and the Hubei Maternal and Child Health Hospital between January 2011 and November 2014.Three hundred and sixteen healthy children from the out-patients of Zhongnan Hospital of Wuhan University during the same period were enrolled as healthy controls.Enzyme linked immunosorbent assay was utilized to measure plasma MT3 protein levels in a dataset of 81 cases and 80 controls,while eight single nucleotide polymorphisms (SNP) located in MT3 gene were genotyped in another greater dataset that included 132 cases and 236 controls by the matrix-assisted laser desorption/ionization time of flight mass spectrometry within the Sequenom platform.Results Plasma MT3 protein level was significantly lower in autistic group compared to healthy controls [(740.0 ± 327.4) ng/L vs (1 007.1 ± 554.3) ng/L,P ＜ 0.001],particularly for boys when stratified by gender (P =0.005).No difference existed in any allele or genotype frequencies between the 2 groups (all P ＞ 0.05).Conclusions The selected autistic children harbored abnormal expression profiles of plasma MT3 protein,which may have no connection with its gene polymorphisms.

Objective To construct 6 kinds of human soluble proliferation-inducing ligand (sAPRIL) cDNA mutants. Methods Six pair primers according to the cDNA sequence of human sAPRIL for different mutants were designed as follows: HEL Th epitope was added at C or N end, the last or first 45 bp DNA was deleted and HEL Th epitope was added, the last or first 90 bp DNA was deleted and HEL Th epitope was added. Different enzyme site was designed at 5′ end of 12 primers, BamH Ⅰ or SalⅠ or HandⅢ. At the same time, 2 pairs of HEL Th epitope sequence were designed and synthesized, and different incision enzyme sticky end sequence was added at the 5′ or 3′ end of each epitope DNA. Mutant sequences were amplified by 6 pairs primers with sAPRIL cloning plasmid as template. The PCR fragment was digested and then ligated with the HEL Th epitope DNA that had been reannealed. The ligation fragments of the head were ligated with digested pQE80 vector fragment again, and were transformed into DH5? competent cell. Recombinant plasmids were identified by digestion and sequencing. Results Six sAPRIL cDNA mutant DNA sequences were obtained by PCR, and the expression plasmids of sAPRIL cDNA mutants including HEL Th epitope sequence were constructed successfully. The result of sequencing proved that mutations generated as designed fully. Conclusion Six sAPRIL cDNA mutant expression plasmids were constructed successfully.

Objective To construct, express, purify and screen immunosuppressive molecule against human soluble APRIL (a proliferation-inducing ligand). Methods The cDNA of soluble APRIL (sAPRIL) was mutated and TEL Th epitope was added. Mutants was expressed by pQE-80L/DH5? system, identified by SDS-PAGE and Western blotting, purified by Ni-NTA resin. Their activity on stimulating the proliferation of Raji cell was detected. Results Four sAPRIL mutants were constructed and expressed as follows: HEL Th epitope was added at C end (Ⅰ); HEL Th epitope was added at N end (Ⅱ); the last 45-bp DNA was deleted at C end and HEL Th epitope was added (Ⅲ); the first 45-bp DNA was deleted at N end and HEL Th epitope was added (Ⅳ). Specific protein bands according to mutant Ⅰ-Ⅳ were detected by SDS-PAGE and Western blotting. Mutant protein was purified by Ni-NTA successfully. Mutants Ⅰand Ⅱ promoted cell proliferation remarkably, and mutant Ⅲand Ⅳnot. Conclusion Four sAPRIL mutants was constructed and expressed successfully. Immunosuppressive molecule against sAPRIL that can not promote cell proliferation was screened out, and it laid a foundation for further study on their immunosuppressive function.

Objective To investigate the role of ribosomal protein S7(rpS7) in the defense of Anopheles dirus against infection. Methods rpS7 was amplified from Anopheles dirus hemocytes with degenerated primers designed according to the conservative region of S7, rpS7 was then cloned using T/A cloning kit and the inserted fragment was sequenced. The difference of the transcript abundance of rpS7 from Anopheles dirus hemocyte among non-blood-fed (N), normal-blood-fed (B) and Plasmodium yoelii infected groups (I) was also analyzed by RT-PCR and gel scanning system at d1, d2, d3, d4, d7 and d11 after blood feeding. Results There is no significant difference of rpS7 signal between the three groups. Conclusion Anopheles dirus S7 can be used as an internal control to study the role of Anopheles dirus related immune factors in Plasmodium infection.

OBJECTIVE To investigate the distribution and drug resistance of the bacteria encountered from the(clinical) infection,and provide reference for the rational use of agents in clinics.METHODS To culture the(specimens) from inpatient and outpatient clinic from Dec 2004 to Mar 2005,and perform the drug (sensitivity) test.RESULTS In 1584 isolated strains,the first four were coagulase negative Staphylococcus((15.6%),) Klebsiella(13.0%),Escherichia coli(10.0%),and Candida albicans(9.9%).The highest resistance of Gram(-negative) bacilli was to ampicillin(86.7%),then to cephalothin(79.5%),and mezlocillin(75.7%);G~-bacilli were(sensitive) to meropenem(2.3%),piperacillin/tazobactam(19.0%),and fosfomycin(29.1%).Gram~+ cocci had drug(resistance) to 18 antibacterials except vancomycin,MRSA was higher than MSSA;(Enterococcus) faecium showed poor sensitive rate to penicillin and erythromycin.CONCLUSIONS We should give more attantion to the Rules on Antibacterials in Clinical Application and use drugs reasonably.