As one of important B vitamins,folate is an essential nutrient for the body and involves in various biochemical and metabolic reactions.Studies have shown that as an important methyl donor in the carbon metabolism cycle,folate has an important impact on pregnancy,pregnancy complications,and birth defects.Further studies have shown that in the folate metabolic pathway,the gene polymorphism and folic acid metabolism of a key enzyme,5,10-methylenetetetrahydrofolate reductase (MTHFR),play an important role in ovarian function.Gene polymorphism and the high homocysteine levels caused by it can lead to the damage of reproductive function and endocrine function,including follicular development,embryonic development,and hormone secretion.Gene polymorphism is also associated with the occurrence and development of ovarian disease and its response to treatment.With the deep understanding of folate metabolism,MTHFR gene polymorphism may become a new genetic marker for predicting the risk of disease and a new target for related gene therapies.

It remains an urgent need to develop effective malaria vaccine for global control of malaria.Application of high technologies such as biotechnology has facilitated the process of vaccine development for malaria.In the past 30 years,a large number of vaccine candidate antigens for malaria have been identified and some of them are currently in clinical trials.Major progress in malaria vaccine development has also been made in China.The PfCP-2.9 blood stage vaccine for malaria has entered clinical studies and some other vaccine candidates including combination malaria vaccine are currently in pre-clinical studies.The availability of various national research programs and international funding has stimulated laboratory and pre-clinical studies of malaria vaccine candidates.It remains a long-term goal to develop a safe and effective malaria vaccine to control and even eliminate the disease in the world,and many issues including malaria immunology and various types of technologies need to be addressed.However,efforts need to be continued toward the goal.

Objective:To synthesize the eba 175Ⅱf2 gene of Plasmodium falciparum and express it in Pichia pastoris in the secreting form. The recombinant protein and PfCP 2.9 protein were used for combined immunization to see if there is antigen competition. Methods: Asymmetric PCR based metho d was utilized to synthesize the 959bp eba 175Ⅱf2 gene. Plasmid containing the synthetic gene was introduced into Pichi a pa storis by electroporation for inducible expression. The recombinant EBA 175Ⅱ F2 p rotein was purified by ion exchange and gel filtration ch romatography. Results: The eba 175Ⅱf2 gene was successfully ex p ressed in Pichia pastoris in the secreting form. The antibody titers in mice immunized with the combined EBA 175ⅡF2 and PfCP 2.9 proteins w ere much higher tha n those with individual protein by ELISA. Conclusion: No antigen competition is found when using the combined EBA 175ⅡF2 and PfCP 2.9 immunization in mice, indicating the potential of combining the 2 antigens for vaccination.

Objective To analyze the imaging features of solitary focal ground-glass opacity nodules (fGGO)by multi-slice spiral CT(MSCT)reconstruction technique in order to improve the differential diagnosis between benign and malignant fGGO and the diag-nosis of early-stage lung cancer.Methods 50 lesions confirmed by pathology were divided into three groups including preinvasive nodules in 19,invasive adenocarcinoma in 10 and benign ones in 21.All CT images were processed by coronal and sagittal recon-struction,maximum and minimum intensity projection (MIP & minIP)and VR.The relationships between fGGO and bronchus were divided into four types:Type Ⅰ with abruptly obstructed bronchus by the fGGO;Type Ⅱ with penetrated and conical inter-rupted bronchus by the fGGO;Type Ⅲ with normal bronchus;and Type Ⅳ with fGGO neighboring the bronchus.In addition,the relationships between fGGO and vessel were divided into three types:Type Ⅰ with normal vessel in or near the fGGO;Type Ⅱ with taper-like narrowed or interrupted one in fGGO;and Type Ⅲ with obstructed one by the fGGO.The clinical data,lesion type,mar-gin,internal structure (air bronchograms/vacuole sign),adjacent structures (vascular convergence/pleural retraction)and the rela-tionships between lesion and adjacent bronchus or vessel were statistically analyzed.Results No statistical differences between three groups were found in the sex of patient and lesion type.A significant difference was found in the age of patients (P=0.005)with less age in benign group than that in preinvasive or invasive adenocarcinoma group.The margin,internal and adjacent structures of the lesions were significantly different (P<0.05).Among the different types of relationship between fGGO and brochus,type Ⅱand Ⅲ were often seen in the preinvasive and the invasive adenocarcinoma groups (the invasive adenocarcinoma often with type Ⅱ), and the type Ⅲ and Ⅳ were in the benign group (benign nodules only with the type Ⅳ).Among the types of relationship between fGGO and vessel,type Ⅱ was seen commonly in the preinvasive group, type Ⅲ often in the invasive adenocarcinoma group,and type Ⅰ only in the benign group.Conclusion The suggested signs with malignant possibility may include the older age,the lesions with lobulated and/or speculated margin,air bronchograms,vacuole sign,pleural retraction around the lesion,vascular convergence sign,and obstructed or cone-shaped narrowed bronchi or vessels in or near the nodules.Thin-section imaging reconstruction techniques help to fully display these signs.

Objective To produce an erythrocytic stage chimeric protein of Plasmodium berghei in Pichia pastoris and evaluate its immunogenicity. Methods The DNA sequences of AMA1 (Ⅲ) and MSP1-19 from P. berghei homologous to the corresponding sequences of P. falciparum chimeric antigen 2.9 (PfCP-2.9) were fused to generate a chimeric gene, designated as PbCP-2.9. The resulting gene was redesigned using Pichia preferential coden usage and expressed in P. pastoris in the secreted form. The recombinant protein was purified by Ni-NTA affinity chromatography. Three groups each with 10 BALB/c mice were im- munized subcutaneously with 20 ?g of purified PbCP-2.9 antigen formulated in Freund’s adjuvant, Montanide ISA720 and Montanide IMS 1 312, respectively. Three control groups each with 10 mice received only adjuvants emulsified with PBS. All the mice received three immunizations at 2-week intervals with the same dose of antigen. Serum samples were collected at preimmunization and one week after each immunization, and were analyzed for specific antibodies by ELISA and reaction with natural P. berghei proteins by IFAT. Results The PbCP-2.9 antigen with Mr 26 400 was successfully expressed in P. pastoris in secret- ed form. The recombinant protein can be recognized by the serum against blood stage parasites of P. berghei. High antibody responses were detected in all three PbCP-2.9-immune groups of mice by ELISA. However, mice immunized with PbCP-2.9 antigen in Freund’s adjuvant produced higher antibody titers than those with PbCP-2.9 antigen in Montanide ISA 206 and Montanide IMS 1312 adjuvants. The mean antibody titer in Freund’s adjuvant was 6.9-fold higher than in Montanide ISA 206 adjuvant and 5.6-fold higher than in Montanide IMS 1312 adjuvant after the second immunization (F=81.06, P

Objective To explore the association between rheumatoid arthritis (RA) with TLR4 Asp299Gly functional variant. Method The literature about RA and TLR4 Asp299Gly functional variant were searched and the meta-analysis was performed. Results Three studies enrolled 718 RA patients and 1392 controls in total. The analysis showed that the TLR4 Asp299Gly functional variant was neither associated with the susceptibility of RA [OR=1.23 (0.67, 2.25), P=0.5]; nor associated with SE or RF [OR=0.67 and 1.02 respectively]; while the TLR4 Asp299Gly functional variant was associated with the age of onset [OR=-4.35 (-7.45, -1.25), P=0.006]. Conclusion The meta-analysis reveals that the TLR4 Asp299Gly functional variant is not associated with the susceptibility of RA, but the onset of RA is earlier in the AA type than those in the AG type. Further prospective corhort studies should be carreied out in the future, especially in oriential people.

Objective To express and evaluate the immunogenicity of ectodomain and its subdomains of Plasmodium berghei apical membrane antigen 1(PbAMA-1).Methods Sequence of PbAMA-1 gene was isolated from the genome of P.berghei,and was redesigned and divided into three overlapped fragments according to its subdomain structure.The codon-optimized DNA fragments of PbAMA-1 were synthesized and inserted into vector pET32a for expression in E.coli and the recombinant proteins were purified by Ni-NTA column,followed by refolding in vitro.Mice and rabbits were immu-nized with the recombinant proteins formulated with Freund adjuvant.Titer of the specific antibodies was detected by ELISA and IFA.The immunized mice were challenged by P.berghei to evaluate protective efficacy in vivo.Results The sequence of the PbAMA-1 gene was shown to be identical to that published before.PbAMA-1 sequence was redesigned via codon optimization and synthesized.Both ectodomain and its subdomains of PbAMA-1 were successfully expressed in E.coli after induction.The proteins were isolated with the purity of more than 90% after Ni column purification and refolding in vitro.Immunization of mice with the recombinant proteins induced high level of specific antibodies.The antibody titer to ectodomain E after the 3rd immunization showed a strong immunogenicity at(34.4?0.15)?10-4.The antibodies interacted with the parasites by indirect fluorescence.The immunized mice were partially protected from the challenge of P.berghei.Conclusion The recombinant PbAMA-1 is highly immunogenic and induces protective immunity against the challenge of P.berghei.

Objective To study the effects of injecting recombinant human granulocyte colony stimulating factor (rhG-CSF) and basic fibroblast growth factor (bFGF) alone or combined on actue myocardial infarction(AMI).Methods AMI models were induced by ligation of the left anterior descending artery.The survived rats were divided into four groups randomly:AMI group (MI),rhG-CSF group (G),bFGF group (B),combined group (GB).Respectively,saline,rhG-CSF,bFGF,and rhG-CSF plus bFGF were injected intraperitoneally 24 h after AMI.Also,sham-operated group (S) was established with only chest-opeaned,without ligation,and no drugs intervention. The white blood cells (WBC) and mononuclear cells (MNC) proportion in peripheral blood were counted 1 week before and 1 week after the intervention,and the number of CD34+ cells was observed with immunohistochemical staining 1 week after AMI in order to compare the situation of mobilization in peripheral blood;the capillary density was evaluated by HE staining both 1 and 4 weeks after AMI;their cardiac fuction was determined in vivo,the infarction size in each group was calculated,and the pathological changes in rat myocardium were observed by HE staining 4 weeks after AMI.Results Compared with MI group,the number of WBC and MNC% in peripheral blood 1 week after AMI in G,B and GB groups were higher(P

Objective To investigate immunogenicity and protection efficacy of the recombinant hypoxanthine-guanine-xanthine (HGXPRT) of Plasmodium falciparum expressed in Pichia pastoris. Methods 35 BALB/c mice were divided randomly into five groups: HGXPRT+ISA720 experiment group, HGXPRT+Freund experiment group, ISA720 adjuvant control group, Freund adjuvant control group, and blank control group. BALB/c mice were subcutaneously immunized three times with the HGXPRT protein formulated by either Freund or ISA720 adjuvants at a three weeks interval. Mice were bled via tail vein at 2 weeks after each immunization. Specific antibodies were detected by ELISA as well as IFAT using cultured parasites. The immunized mice were challenged with 105 P.yoelii 10 days after the third immunization and parasitemia was monitored daily by examining Giemsa-stained thin film. Results Strong immune response was induced by the HGXPRT antigen formulated with the adjuvant. Antibody titers of more than 1∶105 were detected after the third immunization while no specific antibody was detected in the mice immunized with adjuvants only. The antibodies against HGXPRT recognized the cultured parasite by IFAT. Four days after mice were challenged with P.yoelii, high parasitemia appeared in the two control groups, which were 24 h earlier than experiment groups. The mean parasitemia of HGXPRT+ISA720 experiment group(29.3%) was significantly lower than that of control groups (70.0%) (P

Neuromyelitis optica ( NMO) is an idiopathic autoimmune inflammatory disorder of the central nervous system ( CNS) that predominantly affects the optic nerves and spinal cord.It is especially common in Asia. An association between NMO and other autoimmune diseases has been frequently reported, which is different from multiple sclerosis( MS) ,this phenomenon may due to different genetic background between these two diseases.There are reports on different populations to make this clear,which showed human leukocyte antigen( HLA) class II distribu-tion was unique entity different between NMO and MS or healthy controls,this would be a great help to make clear the mechanism underlying NMO pathogenesis.