Objective To study protective immunity effects of co-immunization with P30 DNA vaccine and protein vaccine. Methods Forty-eight 5-6 weeks old BALB/c female mice were divided into four groups (A,B,C,D), 12 mice of each group. In group A (control group) each mouse was immunized with 100 ?g pcDNA3.1 plasmid DNA by intramuscular (i.m.) for three times at week 0,2 and 4; in group B (P30 protein group) each mouse was immunized (i.m.) with 50 ?g rP30+50 ?g CFA for three times at week 0, 2 and 4; in group C (pcDNA3.1-P30 group) each mouse was immunized with 100 ?g pcDNA3.1-P30 plasmid DNA (i.m.) for three times at week 0, 2 and 4; in group D (P30 DNA+rP30 co-immunization group) each mouse was immunized with 100 ?g pcDNA3.1-P30 plasmid DNA (i.m.) for two times at week 0, 2 and immunized by subcutaneous with 50 ?g rP30+50 ?g CFA at week 4. Each mouse was infected with 100 tachyzoites of Toxoplasma gondii RH strain four weeks later after last immunization. The anti-P30 antibodies were detected with ELISA before the challenge. Results The P30 DNA vaccine was successfully constructed. High titers of anti-P30 antibodies were induced in each mouse immunized with DNA vaccine. The protective trial proved that there was no significant difference between control group and experimental group though the survival time of mouse from experimental group had been prolonged. Conclusion The P30 DNA vaccine could induced high titers of anti- P30 antibodies in immunized mice, and it may be a potential DNA vaccine candidate.

Adolescent ; Female ; Humans ; Immunoglobulins, Intravenous ; administration & dosage ; Immunosuppressive Agents ; administration & dosage ; Polyradiculoneuropathy, Chronic Inflammatory Demyelinating ; diagnosis ; therapy

Objective To research the significanec of genes bel-2 and bax in chondrocyte apoptosis in experimental osteoarthritis in rabbits.Method Twelve New Zealand rabbits divided into two groups at random:model group and control group.Model group with osteoarthritis was duplicated by immobilizing with gypsum at extension position in right hind limb of rabbits.Rabbits of model group were executed after 6 weeks and chondrocytes were taken.Positive expression rate of bcl-2 and bax mRNA was measured by immunohisto- chemistry and Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL).Results Positive expression rate of mRNA in bel-2 and bax of model group was obviously more than control group(P＜0.05).According to immunohistochemistry,and grey grade of positive expression of protein in bcl-2 and bax of chondrocytes in model group was lower and positive expression rate was gigher compared with control group (P＜0.05,P＜0.01).bcl-2 and bax of model group were lower than control group(P＜0.05).Conclusion Genes bcl-2 and bax participate and thus accelerate chondrocyte apoptosis of osteoarthritis together.

Three new compounds, including a naphthoquinone, a reduced naphthoquinone derivative naphthalenone, and a tricarboxylic acid, along with five known naphthalenone derivatives were isolated from ethyl acetate extract of rice fermentation products of the fungus Pleosporales sp. by multiple column chromatographic methods, including Sephadex LH-20 gel column chromatography, silica gel column chromatography, reversed-phase HPLC, and chiral chromatography. Their structures were elucidated by MS, NMR, and specific rotation spectroscopic analyses as well as ECD calculations. Three novel compounds were named as pleospathone A (1), pleospathone B (2), and pleosporalic acid A (3). Five known compounds were separately identified as (3S,4R)-3,4,8-trihydroxy-6-methyl-3,4-dihydronaphthalen-1(2H)-one (4), (4R)-3,4-dihydro-4,6,8-trihydroxy-1(2H)-naphthalenone (5), (-)-scytalone (6), (3S,4S)-3,4-dihydro-3,4,6,8-tetrahydroxy-1(2H)-naphthalenone (7), and cis-4-hydroxyscytalone (8). Compounds 4-8 were isolated from the Pleosporales fungi for the first time. Compound 1 shows inhibitory activities against human cervical carcinoma cell line HeLa and murine leukemia cell line P388 with the IC₅₀ values of 78.93 and 98.80 μmol·L^-1, respectively.

Objective To discuss the anatomic features, clinical presentations, diagnosis,differentiations and treatments of congenital fourth branchial anomaly(CFBA). Methods The clinical data of 8 patients with CFBA were retrospectively analyzed. Results Of the 8 patients aging from 27 to 300 months(median age: 114 months), 4 male and 4 female; 3 untreated previously and 5 recurrent. All lesions, including 1 cyst, 3 sinus (with internal opening) and 4 fistula, located in the left necks. Three patients presented acute suppurative thyroiditis, 4 deep neck abscesses, and 1 neck lump. Preoperative examinations included barium esophagogram, direct laryngoscopy, ultrasonography, CT, MRI, and so on.The principles of managements were adequate drainage, infection control during acute period and radical surgery during quiescent period. Classic surgical approach consisted of complete excision of branchial lesions, dissection of recurrent laryngeal nerve and partial thyroidectomy. Selective neck dissection was applied in recurrent cases to extirpate branchial lesions, scarrings and inflammatory granuloma.Postoperatively, 1 case was with local incision infection which healed by wound care; 1 case was with temporary vocal cord paralysis which completely recovered 1 month after operation. No recurrence was found in all of 8 cases with follow-up of 13 to 42 months (median: 21 months). Conclusions CFBA relates closely anatomically with recurrent laryngeal nerve and thyroid grand. The barium esophagogram and direct laryngoscopy are the most useful diagnostic tools. CT and MRI are all beneficial to the diagnosis of CFBA.The treatment key to CFBA is the complete excision of lesion during a quiescent period after inflammatory control, together with the dissection of recurrent laryngeal nerve ,partial thyroidectomy and partial resection of lamina of thyroid cartilage (if necessary), which all can decrease the risk of complications and recurrence.For recurrent cases, selective neck dissection is a safe and effective surgical procedure.

Alveolar epithelial type II (AT II) cells are essential for lung development and remodeling, as they are precursors for type I cells and also produce other non-repair cells (fibroblasts). Progenitor cells are believed to possess capability of multi-potent transdifferentiation, which is closely related to the niche, suggesting the importance of establishment of a lung progenitor cell niche model. We hypothesized that pulmonary surfactant-associated protein A (SPA) suicide gene system would cause AT II cell to kill itself through apoptosis and leave its niche. In vitro, the recombinant adeno-associated virus vectors-SPA-thymidine kinase (rAAV-SPA-TK) system was established to get targeted apoptotic AT II cells. The apoptosis of AT II cells was detected by using MTT. The results showed that cloned SPA gene promoter had specific transcriptional activity in SPA high expression cells, and SPA high expression cells (H441) transfected with TK gene had higher sensitivity to ganciclovir (GCV) than SPA low expression cells (A549). In vivo, increased apoptosis of AT II cells induced by GCV in rAAV-SPA-TK system was observed by TUNEL. Finally, the successful packaging and application of rAAV-SPA-TK system provide experimental basis to get specific lung progenitor cell (AT II) niche in vitro and in vivo.

OBJECTIVETo explore the effect of basic fibroblast growth factor 1 (bFGF1) during embryonic development on hematopoiesis, to study the expression of FGF1, vascular endothelial growth factor receptor (KDR), CD133, CD34 and transcription factors Ihh, SCL, GATA-1, GATA-2 and PU. 1 in the yolk sac, and to learn about the role and relationships of FGF1, hematopoietic cells and transcription factors during embryonic hematopoiesis.

METHODS10 microm sections and total RNA were prepared from 107 human embryos aged 3-12 weeks. Immunohischemical SP staining and RT-PCR were performed.

RESULTSThe yolk sac blood islands of human 3 approximately 12 weeks embryos consisted of peripheral vascular endothelial cells and central hematopoietic cells. The expression of FGF1 was firstly found in visceral mesoderm around periphery of yolk sac blood island at day 16, while was little inside it. KDR was not or lowly expressed and CD34 and CD133 were not expressed then. The expression increased, gray value decreased and staining enhanced at day 21. Strong staining of CD34+, CD133+ and KDR+ cells were found in blood island and mesoderm at day 30, their gray values changed from 156 +/- 16, 173 +/- 18 and 160 +/- 14 to 53 +/- 7, 52 +/- 6 and 69 +/- 8 respectively. FGF1 expression was strong positive, the gray value declined dramatically from 161 +/- 13 to 40 +/- 5. Some positive cells formed vessel-like structure along the periphery of blood island. Moderate expression of CD34+, CD133+, KDR+ cells increased at day 45, the cells aggregated into mass in blood island and FGF1+ cells did the same in blood island, while little in mesoderm. Its gray valve was increased. After 7 weeks, CD133+, KDR+, CD34+ cells significantly decreased their gray values increased, the staining became week. FGF1 was weakly expressed in yolk sac and its gray value increased to 179 +/- 22. RT-PCR showed Ihh, SCL, GATA-1 and GATA-2 were expressed at different time in yolk sac. PU. 1 were not expressed at day 16, and then expressed.

CONCLUSIONSThe hematopoietic properties of yolk sac may be dependent on signaling through FGF receptors and FGF1 plays an important role in hematopoietic stem cell homeostasis. The FGF pathway regulates primitive hematopoiesis by modulating transcription factors such as Gata1 expression level and activity.

Embryo, Mammalian ; metabolism ; physiology ; Fibroblast Growth Factor 1 ; metabolism ; Hematopoiesis ; Humans ; In Vitro Techniques ; Yolk Sac ; metabolism ; physiology

Objective To investigate the application of SIEMENS Ysio DR amplification in the radiation dose reduction. Methods Totally 160 patients with similar heights and body shapes were divided equally into A,B,C and D groups,and then underwent radiological examinations with SIEMENS Ysio DR from September 2014 to May 2016.A and B groups went through right hand AP oblique examination with the same kV while different mAs, and C and D groups had chest posteroanterior examination with different kV and the same mAs. Image quality was regulated with the amplification coefficient, and then evaluated with double-blind method by two senior radiologists.Statistical analysis was executed over the values of mAs in A and B groups and those of kV in C and D groups. Results A group had the mAs value being(2.26 ±0.16)mAs, which was significantly different from that of B group [(4.05±0.19)mAs] (P<0.05);while C group had kV value being (104.79±6.39)kV, which was statistically different from that of D group [(83.48±3.67)kV] (P<0.05). Satisfactory image were obtained by regulation with amplication coefficient.There was no obvious difference between the image quality in either A and B groups or C and D groups(P>0.05).Based on the principle that low mAs resulted in low radiation dose and high kV lead to low effective dose,A group had the radiation dose lower than those of B group,and C group had the effective dose lower than those of D group. Conclusion SIEMENS Ysio DR has the photographing parameters regulated according to different sites and the amplification coefficient for enhancing image quality, which decreases X-ray radiation dose and effective dosage with the diagnosis results unaffected.

OBJECTIVETo study the effect of Hath1 gene transfer on the proliferation of colonic cancer cells in vitro.

METHODSThe recombinant plasmid pcDNA3.1(+)-Hath1 was transfected into HT29 colonic cancer cells, and 3 positive cell clones were randomly selected to test the levels of Hath1 mRNA, Muc2 mRNA, Hath1, Muc2, cyclin D1 and p27 by quantitative real-time RT-PCR and Western blotting. The proliferation of the transfected HT29 cells was observed by means of colony formation assay and xenograft growth in nude mice.

RESULTSHath1 significantly down-regulated of cyclin D1 and up-regulate of p27 expressions and inhibited the proliferation of HT29 cells.

CONCLUSIONHath1 gene may be an anti-oncogene in colon carcinogenesis.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; pathology ; Cyclin D1 ; genetics ; metabolism ; Gene Transfer Techniques ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Mucin-2 ; genetics ; metabolism ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; RNA, Messenger ; genetics

OBJECTIVETo study the role of the synchronized springy lengthening apparatus for the tibia and calcaneal tendon designed by the author in preventing the clubfoot of secondary to the Ilizarov tibia lengthening.

METHODSBased on the Ilizarov tibia lengthening apparatus, a special synchronized springy lengthening apparatus for the tibia and calcaneal tendon was designed. The tibial was made of distal and proximal 2 rings respectively and 4 threaded rods, and the calcaneal was made of a half ring, 2 hinges and a threaded rod with spring. The half ring was fixed to the calcaneus by 2 crossed wires. The fracture tibia and fibula, ankle joint, talocalcaneal joint were attached to the apparatus. At the same time of tibia lengthening, the soft tissue was simultaneously stretched, the ankle joint could move, and the leg could bear weight. If the clubfoot angle was larger, the percutaneous fasciotomy of calcaneal tendon was performed; if the angle was less than 20 degrees, the pes deformities were corrected only by the stretch of calcaneal tendon.

RESULTSSeventy-seven patients' tibia were lengthened averagely 4.6 cm, with an average speed of 0.7 mm/d. The healing made tibia lengthened, and the index was 1.35 months/cm. There were not the secondary varus and valgus deformities and clubfoot in all the patients. The clubfoot with 100-400 angle of the 16 patients were corrected after tibia lengthening.

CONCLUSIONSThe new apparatus coincides with the biomechanical principle and can effectively prevent the secondary deformities of foot such as clubfoot, talipes varus and valgus after tibia lengthening procedure.

Adolescent ; Adult ; Bone Lengthening ; instrumentation ; methods ; Child ; Equipment Design ; Female ; Humans ; Leg Length Inequality ; surgery ; Male ; Treatment Outcome