Objective To construct and identify the transgenic plant vector recombinant pBI-Eg95 plasmid of Echinococcus granulosus. Methods Total RNA was extracted from hydatid cyst protoscoleces of Echinococcus granulosus after sonication. A couple of specific primers were designed on the basis of known sequences of Eg95 gene. The desired gene was amplified by PCR technique from the cDNA, and then was cloned into the plant expression vector pBI121 to construct the recombinant pBI-Eg95 plasmid. The recombinant plasmid was electroporated into Agrobocterium tumefaciens (At) LBA4404 strain. The positive recombinant clones were confirmed by restriction endonuclease digestion and characterized by PCR. Results For RT-PCR, a specific band around 471 bp was amplified. The result of DNA sequencing of Eg95 showed the identity with the published sequence. The same band was obtained by restriction endonuclease digestion and PER from the plasmids of positive recombinant At(rAt). Conclusions The recombinant pBI-Eg95 plasmid was successfully constructed, and it provides the basis to further research of the transgenic plant vaccine of Echinococcus granulosus.

Neutralizing antibodies are recognized to be one of the essential elements of the adaptive immune response that must be induced by an effective vaccine against HIV. However, only a limited number of antibodies have been identified to neutralize a broad range of primary isolates of HIV-1 and attempts to induce such antibodies by immunization were unsuccessful. The difficulties to generate such antibodies are mainly due to intrinsic properties of HIV-1 envelope spikes, such as high sequence diversity, heavy glycosylation, and inducible and transient nature of certain epitopes. In vitro neutralizing antibodies are identified using "conventional" neutralization assay which uses phytohemagglutinin (PHA)-stimulated human PBMCs as target cells. Thus, in essence the assay evaluates HIV-1 replication in CD4+ T cells. Recently, several laboratories including us demonstrated that some monoclonal antibodies and HIV-1-specific polyclonal IgG purified from patient sera, although they do not have neutralizing activity when tested by the "conventional" neutralization assay, do exhibit potent and broad neutralizing activity in "unconventional" ways. The neutralizing activity of these antibodies and IgG fractions is acquired through post-translational modifications, through opsonization of virus particles into macrophages and immature dendritic cells (iDCs), or through expression of antibodies on the surface of HIV-1-susceptible cells. This review will focus on recent findings of this area and point out their potential applications in the development of preventive strategies against HIV.

OBJECTIVETo explore the expression patterns of the chemokine CXCL12 and its receptor CXCR4 in human sperm.

METHODSWe collected semen samples from 10 fertile men, performed density gradient centrifugation, and then determined the expressions of both CXCL12 and CXCR4 in the sperm by RT-PCR and immunofluorescence staining.

RESULTSRT-PCR revealed the mRNA expressions of CXCL12 (0.641 +/- 0.180) and CXCR4 (0.464 +/- 0.100) in the sperm. However, only CXCR4 rather than CXCL12 was expressed at the protein level, and the positive staining for CXCR4 was observed mainly in the posterior part of the acrosome.

CONCLUSIONCXCL12 and CXCR4 are involved as important molecules in regulating the function of human sperm.

Acrosome ; metabolism ; Centrifugation, Density Gradient ; Chemokine CXCL12 ; metabolism ; Humans ; Male ; Receptors, CXCR4 ; metabolism ; Signal Transduction ; Spermatozoa ; metabolism

Recombinant expression vector pcDNA3-MCPCD59DP containing human membrane complement regulatory proteins(hCRPs) MCP and CD59 cDNA was constructed successfully by using two independent promoters.After transfected into NIH3T3 cells with calcium phosphate-DNA precipitate method,NIH3T3 pcDNA3-MCPCD59-DP transfectants were obtained by G418 selection.Extraneous genes integration was identified by PCR.The co-expression of human CD59 and MCP at both mRNA and protein levels was confirmed by using RT-PCR and Western blot analysis.Human MCP and CD59 cDNA were integrated in NIH3T3 pcDNA3-MCPCD59-DP genomic DNA after continuous 30 times passages,indicating that NIH3T3 pcDNA3-MCPCD59-DP were stable cell lines.Human complement-mediated cytolysis assays showed that NIH3T3 cells transfected stably with pcDNA3-CD59,pcDNA3-MCP,and pcDNA3-MCPCD59-DP were protected from C-mediated damage and co-expressed human MCP and CD59 provided more excellent protection against C-mediated attack as compared with either CD59 or MCP expressed alone.The dicistronic vector represents an effective and efficacy strategy to overcome C-mediated damage and has potential therapeutic value for effectively controlling complement activation and finally for preventing hyperacute rejection in clinical gene therapy.

Objective To cultivate and identify the transgenic affalfa containing Echinococcus granulosus Eg95 gene. Methods The alfalfa plants were transformed by co-cultivating alfalfa cotyledons via recombinant Agrobacterium tumefaciens LBA4404 harboring pBI-Eg95. The transgenic alfalfa explants were selected by kanamyein after calli formation, shoots and roots regeneration in the selective medium, the seedlings of transgenic plants were obtained which were finally transplanted into pots containing nutrient soil. After 2-3 months growth, the complete transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene were obtained. To identify the transgenic alfalfa plants, the total DNA, RNA and leaf protein were extracted from fresh leaf tissue of the transgenic alfalfa plants and confirmed by PCR, RT-PCR, SDS-PAGE and Western blot assay. Results A specific band around 471 bp was amplified by PCR with total DNA, and the same band was obtained by RT-PCR with total RNA, which confirmed that the Eg95 gene was stably integrated into the transformed alfalfa genome. SDS-PAGE analysis showed that the relative molecular mass(Mr) of the expressed protein was about 16.5×103, consistent with the Eg95 protein, and the level of Eg95 expression was up to 0.06% of total soluble leaf protein by Bio-Rad Quantity one assay. Western blot verified the expressed protein was reactive with the sera of mice infected with Echinococcus granulosus. Conclusion The transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene are successfully cultivated.

Diagnosis, Differential ; Erythema ; diagnosis ; etiology ; pathology ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Skin ; blood supply

Objective To determine the safety, pregnancy outcome and the affect on neonates in pregnant patients with systemic lupus erythematosus(SLE).Methods Sixty-two pregnant patients with SLE were evaluated retrospectively from 1999 to 2009 in our hospital. These patients were divided into two groups:selective pregnancy group and nonselective pregnancy group. The pregnancy outcomes, fetal outcomes, and lupus activity during pregnancy were compared between the two groups. The children of the SLE patients were followed up. Results There were 43 patients in the selective pregnancies group and 19 patients in nonselective pregnancies group. In the selective pregnancies group, lupus flare occurred in 10 pregnancies(23%), 35(81%)had a live birth, 7 had low birth weight infants and 7 had premature delivery; however, in the nonselective pregnancies group, lupus flare occurred in 16 pregnancies(84%), 13(68%) had abortion,6 had a live birth, but all neonates were low birth weight infants. The rates of lupus flare and pregnancy loss in the nonselective pregnancy group were higher than those of the selective pregnancy group(P＜0.05). None of the 22 children had SLE during the follow-up period. Conclusion Both selective and nonselective pregnancy may adversely affectmaternal and fetal outcomes, but patients with selective pregnancy have better outcomes either in lupus flare or maternal and fetal outcomes compared with those of the nonselective pregnancy.

This paper summarize the practice in the teaching reform of microbiology experiment in recent years. We identify the main contents of experimental teaching systems and pay much more attention to peo-ple-oriented. Through the reform of teaching and assessment methods,students are trained to cultivate their practical ability and spirit of innovation.

ObjectiveTo observe the rehabilitative effect of patients with shoulder-hand syndrome after stroke by manipulation treatment. MethodsThe patients with shoulder-hand syndrome were randomly divided into two groups, manipulation group (180cases) and control group (128 cases). Patients in the manipulation group were regularly given a passive quantitative movement on shoulder, elbow and hand joints,while patients in the control group were irregularly given a passive movement or ordered to perform an autonomic movement. The signs and symptoms of patients in these two groups were not much different. The rehabilitative effects were compared 3 months later. ResultsSigns and symptoms in the manipulation groups improved much better than that of the control group. Conclusions The manipulation treatment for the post-stroke patients with shoulder-hand syndrome is the method that is simple, effective and easy to perform.

Objective To investigate the clinical effect of pulmonary rehabilitation therapy including respiratory exercise and vibration expectoration on patients with pulmonary infection after abdominal surgery.Methods A retrospective case control study was conducted.Seventy-six patients with pulmonary infection after abdominal surgery admitted to the First Affiliated Hospital of Hunan Normal University from September 2015 to September 2016 were enrolled.According to whether accept the pulmonary rehabilitation therapy or not,the patients were divided into two groups.In the control group (n =35),the convemional expectoration method was adopted.The patients in pulmonary rehabilitation group (n =41) received both methods of the control group and pulmonary rehabilitation treatment,including respiratory exercise (effective cough,lip reduction breathing),respiratory exercise device (respiratory exerciser tri-ball),and vibrated expectoration.The 24-hour sputum volume,degree of comfort,inflammatory and pulmonary function parameters,and recovery situation were recorded in the two groups.Results ① There were no significant differences in the parameters of inflammation and pulmonary function before treatment between the two groups.After treatment,the white blood cell (WBC) and C-reactive protein (CRP) in both groups were significantly decreased,and the forced expiratory volume in 1 second (FEV1) and FEV1/forced vital capacity (FVC) were significantly increased.The above changes in pulmonary rehabilitation group were more significant than those of the control group [WBC (× 109/L):11.12 ± 2.88 vs.13.42 ± 2.62 at 3 days,8.22 ± 1.48 vs.9.27 ± 1.92 at 5 days;CRP (mg/L):13.47 ± 4.77vs.16.03±4.94 at 3 days,9.69±1.56 vs.11.77±1.41 at 5 days;FEV1 (L):2.48±0.14 vs.2.29±0.16 at 3 days,FEV1/FVC:0.78±0.04 vs.0.75±0.04 at 3 days;all P ＜ 0.05].② The 24-hour sputum volume within 3 days of pulmonary rehabilitation group were significantly higher than that of the control group (mL:30.51 ± 4.15 vs.18.30 ± 3.64at 1 day,31.08±3.22 vs.20.37±3.20 at 2 days,29.03±2.55 vs.19.03±2.51 at 3 days,all P ＜ 0.01].③ In the pulmonary rehabilitation group,the recovery time of pulmonary infection symptoms (days:5.44 ± 1.45 vs.6.20 ± 1.55),the days of antibiotic use (days:12.61 ± 3.15 vs.15.03 ± 3.78),the time of getting out of the bed (days:4.05 ± 0.74vs.4.51±0.89),and the hospital days (days:19.95±3.90 vs.22.00±4.42) were significantly shorter than those of the control group (all P ＜ 0.05),and the degree of comfort was significantly better than that of the control group (comfort score:2.71 ±0.90 vs.2.14±0.91,P ＜ 0.01).Conclusion The application of pulmonary rehabilitation including respiratory exercise and vibration expectoration in abdominal surgery patients with pulmonary infection can promote recovery,and it has a good clinical and practical application value.