Objective To clone and express the extracellular portion of mouse Fcγreceptor Ⅱ-b ( FcγRⅡb) and to analyze the functions of the expressed proteins in a mouse model of systemic lupus ery-thematosus ( SLE) .Methods The gene fragment encoding FcγRⅡb was amplified by PCR, and then in-serted into the prokaryotic expression vector pET-32a(+) to construct the recombinant expression plasmid pET-FcγRⅡb.The expression plasmid was identified with restriction enzymes and then sent to the Shanghai Bio-Engineering Co.LTD for further sequencing analysis.The transformed Escherichia coli ( E.coli) BL21 ( DE3) strains carrying the recombinant expression plasmid pET-FcγRⅡb were induced by isopropylβ-D-1-thiogalactoside ( IPTG) .The expressed fusion proteins were analyzed by Western blot assay and purified with purification kits.The immune complex ( IC)-binding ability of FcγRⅡb was measured by ELISA.MRL/lpr mice with SLE in both the prevention (12 weeks old, n=40) and the treatment groups (19 weeks old, n=40) were randomly divided into four groups including 60 μl (4.8 μg) treatment group, 120 μl (9.6 μg) treatment group, 180μl (14.4μg) treatment group and PBS treatment group with 10 mouse in each group. The MRL/lpr mice with SLE were injected with the fusion protein through tail vein once a week for four con-secutive weeks.Serum samples were collected from each mouse after one week of observation.The levels of FcγRⅡb in soluble form in mice form both the prevention and treatment groups as well as the levels of anti-double stranded DNA antibodies were detected by ELISA.Results The gene encoding FcγRⅡb was ampli-fied and the recombinant expression plasmid pET-FcγRⅡb was successfully constructed.The recombinant proteins were expressed in the prokaryotic expression system, and then successfully purified.The recombi-nant proteins could bind to IC.Compared with the corresponding PBS control group, the levels of FcγRⅡb in soluble form were increased in mice from both prevention and treatment groups after treating with various concentrations of the recombinant protein (P<0.05).Significant differences in the levels of FcγRⅡb were found among mice of the same age after treating with different concentrations of the recombinant protein ( P<0.05).Compared with the corresponding PBS control group, the levels of anti-double stranded DNA anti-bodies were decreased in mice from both prevention and treatment groups after treating with various concen-trations of the recombinant protein (P<0.05).The levels of anti-double stranded DNA antibodies were grad-ually decreased along with the increasing dosage of protein (P<0.05).Conclusion The extracellular por-tion of murine FcγRⅡb in soluble form was successfully expressed.The recombinant proteins played a cer-tain role in the prevention and treatment of SLE in a mouse model.

Objective:To analyze the effects of silver ion on the integration frequency of the class 1 integron in Escherichia coli ( E. coli) BL21(DE3) host. Methods:Two recombinant plasmids, pUCINT and pACINAD, were successively transformed into E. coli BL21 (DE3) to construct HS2 strains. Three experimental groups were set up using 0.3 μg/ml, 0.6 μg/ml and 0.8 μg/ml silver ion LB liquid medium, while control group used common LB liquid medium. Silver ion was supplied by silver nitrate and HS2 strains were cultured at 37℃ for 24 h. The copy number of cointegrates and the total copy number of integrons in each group were detected by real-time polymerase chain reaction (qPCR), and the ratio of them was the integration frequency. Changes in the integration frequency were analyzed by three independent phenotypic screening method and the protein expression in HS2 strains was analyzed by mass spectrometry. Results:The integration frequency in HS2 strains in the control group and three experimental groups (0.3 μg/ml, 0.6 μg/ml and 0.8 μg/ml silver ion) was 1.79×10 -5 (1.44×10 -5, 3.13×10 -5), 2.07×10 -5 (1.49×10 -5, 2.67×10 -5), 2.25×10 -6 (1.47×10 -6, 4.54×10 -6) and 1.69×10 -6 (0.22×10 -6, 3.08×10 -6), respectively. The integration frequency in the 0.6 μg/ml and 0.8 μg/ml silver ion groups was significantly lower than that in the control group ( P<0.01), but there was no significant difference between the 0.3 μg/ml silver ion group and the control group. Results of three independent phenotypic screening method were consistent with those obtained by qPCR. Mass spectrometry analysis showed that there were differences in protein expression in HS2 strains between the control group and the experimental groups. Conclusions:Silver ion at a certain concentration had an inhibitory effect on the frequency of drug resistance gene cassette captured by bacterial integron.

The change in serum laminin (LN) level and its clinical significance in epithelial ovarian tumor were investigated. The LN levels in serum and ascites samples from 69 patients with epithelial ovarian tumor and 42 cases as control group before and after operation were analyzed by radioimmunoassay. The results showed that the serum LN levels in the patients with malignant tumors (157.85 +/- 14.37 ng/ml) were significantly higher than that in the control group (125.14 +/- 7.03 ng/ml) and in the patients with benign tumors (128.36 +/- 8.75 ng/ml) (both P < 0.01) before operation. The serum LN levels in the malignant group were decreased significantly after operation as compared with those before operation (P < 0.05). The serum LN levels in low-differentiated tumors was higher than those in moderate-differentiated tumors and high-differentiated tumors (P < 0.05). The LN levels in ascites (172.94 +/- 15.26 ng/ml) was significantly higher than in serum (161.34 +/- 6.59 ng/ml) (P < 0.05) in malignant tumors. The serum LN levels in the patients with lymph node metastasis (165.41 +/- 19.91 ng/ml) was obviously higher than those without lymph node metastasis (152.35 +/- 10.34 ng/ml) (P < 0.05). It was concluded that LN levels in serum and acistis were remarkably increased in malignant epithelial ovarian tumors, suggesting that LN might be one of important diameters reflecting tumor biological characteristics.
Ascitic Fluid/*metabolism ; Carcinoma/blood ; Carcinoma/metabolism ; Laminin/*blood ; Laminin/metabolism ; Ovarian Neoplasms/*metabolism ; Tumor Markers, Biological/*blood ; Tumor Markers, Biological/metabolism

Brain-computer interface (BCI) system is a system that achieves communication and control among humans and computers and other electronic equipment with the electroencephalogram (EEG) signals. This paper describes the working theory of the wireless smart home system based on the BCI technology. We started to get the steady-state visual evoked potential (SSVEP) using the single chip microcomputer and the visual stimulation which composed by LED lamp to stimulate human eyes. Then, through building the power spectral transformation on the LabVIEW platform, we processed timely those EEG signals under different frequency stimulation so as to transfer them to different instructions. Those instructions could be received by the wireless transceiver equipment to control the household appliances and to achieve the intelligent control towards the specified devices. The experimental results showed that the correct rate for the 10 subjects reached 100%, and the control time of average single device was 4 seconds, thus this design could totally achieve the original purpose of smart home system.
Brain-Computer Interfaces ; Electroencephalography ; Evoked Potentials, Visual ; Humans ; Microcomputers ; Wireless Technology

Objective To establish a system for detecting integration frequency of antibiotic resist-ante integron.Methods We cloned integron and aadA2 gene cassette into different sites of plasmid pACYC 184,and the plasmid was transformed into E.coli BL21(DE3)containing plasmid overexpressing integrase.The positive clone was cultured overnight and then was spread on LB agar plate with or without streptomycin respectively,and with appropriate amount of bacteria.Clones after cultured overnight were counted to detect the integration frequency.Meanwhile we used positive clones in LB agar plate containing streptomycin as templates to carry out PCR.The purified PCR products were sequenced to identify the integration sites.Re-suits The integration frequency of integron capturing aadA2 gene cassette in BL21(DE3) host was 1.1 x 10-3 mainly at attI site.Conclusion This system can be used to detect integration frequency.

Objective To observe the osteogenic effect of tissue-engineered bone constructed by poly-L-lysine-demineralized bone matrix (PLL-DBM) enriched bone marrow stem cells in the space of goat transverse process bone fusion model and explore a new tissue-engineered bone construction method. Methods PLL was used to decorate goat DBM to prepare a matrix material (PLL-DBM). The osteo-genic effect of tissue-engineered bone constructed by PLL-DBM enriched bone marrow cells ( Group Ⅰ A) was detected in goat lumbar intertransverse graft bone model; autogenous iliac bone (Group Ⅰ B), DBM enriched bone marrow (Group Ⅱ C) and DBM (Group Ⅱ D) were used as controls. The osteogenesis of the bones in the fused segments of four groups were compared and evaluated by X-ray, three-dimensional CT, CT value testing and biomechanical testing. Results The results of X-ray showed that the fusion ranges in groups ⅠA and ⅠB were basically the same, which were significantly wider than that in Group Ⅱ, with no fusion detected in Group Ⅱ D. The CT value was (696.76±10275) HU in Group Ⅰ A and (766.03±69.24) HU in Group B, which were significantly higher than that in Group Ⅱ C (P <0.05), but there was no statistical difference in CT value between Groups Ⅰ A and Ⅰ B (P > 0.05). The CT val-ue in Group Ⅱ C was significantly higher than in Group ⅡD (P <0.01). There was no statistical differ-ence between Groups Ⅰ A and Ⅰ B in the maximum load and bending strength (P > 0.05). The maxi-mum load and bending strength in Groups Ⅰ A and Ⅰ B were significantly higher than that in Group Ⅱ C (P < 0.05), and the two indices in Group Ⅱ C were significantly higher than that in Group Ⅱ D (P <0.01). Conclusion Tissue-engineered bone constructed by PLL-DBM enriched bone marrow cells is an ideal tissue engineered bone and its osteogenic potential is similar to that of autologous bone.

Objective To explore on FK506 promoting proliferation of Schwann cells in vitro and NGF of Schwaun cells secreted highly by itself. Methods Purified Schwann cells divide into six groups:group A (control group) DMEM/F12 contained 10% calf bloodserum; group B contained 0.1 ng/ml FK506; group C contained 0.5 ng/ml FK506; group D contained FK506:1.0 ng/ml;group E contained FK506:5.0 ng/ml; group F contained FK506:10 ng/ml. Morphology of Schwann cells were oboyrved by invert microscope and evaluated Schwann cells in immunocytochemistry staining with anti-S-100. The best concentration of FK506 who promoted proliferation of Schwann cells by MTT. Cell cycle of Sehwarm cells were determined by flow cytometry. The level of NGF in the conditioned media was determined by an enzyme-linked immunoadsordcnt assay after 72 h. Results Group C was the best concentration which promoting proliferation of Schwann cells among 5 groups, when the concentration 1.0 ng/ml FKS06 to promote Schwann cell proliferation gradually weakened. Detected by flow cytometry showed that: containing 10% fetal DMEM/F12 bovine serum for 24 h,72 h and 48 h after Schwann cells in S phase percentage were 27.8%,39.3% and 58.4% in the 0.5 ng / ml FK506 for 24 h,72 h and 48 h after Schwann cells S percentage period were 54.2% ,60.3% and 94.6%. S phase of the latter than the former in 24 h,72 h and 48 h, respectively higher: 26.4% and 21% and 36.2%. FK506 detected by ELISA promote Schwann cell proliferation after the expression of NGF in the experimental study found: 0.5 ng/ml FK506 for 72 h after the Schwann cells secreted by the NGF as high as 0.188 ng/ml, rcspectiveIy. Conclusion FKS06 can promote proliferation of Schwann cells at early time in vitro and Schwann cells' good situation is highly kept to secrete NGF.

Objective To assess roles of vascular endothelial growth factor(VEGF)and plateletderived growth factor(PDGF)in the mechanisms of lymphangiogenesis in epithelial ovarian carcinoma.Methods (1)Expression of Proxl,a newly described lymphatic endothelial cell nucleus marker,VEGF-A,VEGF-C,VEGF-D and PDGF-A,PDGF-B,PDGF-C,PDGF-D were detected bv RT-PCR in SKOV3 cell line and in 90 ovarian tissue samples,included 15 benigh tumors 10 borderline tumors, 45 malignant tumors and 20 normal ovarian samples.(2)Expression levels of Proxl,VEGF-A,-C,-D and PDGF-A,-B,-C,-D were detected in 90 ovarian tissue sample mentioned above by real-time quantitative PCR(RTQPCR).Resuls (1)Proxl was expressed in ovarian samples mentioned above,while not detected in SKOV3 cell. VEGF-A,-C,-D and PDGF-A,-B,-C,-D were found in SKOV3 cell and various ovarian tissues.(2)Expression levels of Proxl(2.2±1.3,P＜0.01),VEGF-A(3.5±1.5,P＜0.01),VEGF-C(19±14.P＜0.01),VEGF-D(3.0±1.8,P＜0.01)and PDGF-A(3.3±3.3,P＜0.05),PDGF-C(6.9±4.6,P＜0.01)in malignant group were found to be significantly higher than those in borderline group and benign group.(3)The expression levels of Proxl,VEGF-A and PDGF-A were significantly greater in samples from the patients with lymph node metastasis(Proxl:3.0±1.4,VEGF-A:4.1± 1.7,PDGF-A:4.9±4.1),peritoneum metastasis(Proxl:2.8±0.9,VEGF-A:4.0±1. 8,PDGF-A:4.5±4.0)and in stage Ⅲ-Ⅳ(Proxl:2.6±1.3,VEGF-A:4.0±1.4,PDGF-A:4.1±3.7)than those without lymph node metastasis,without peritoneum metastasis and in stage Ⅰ-Ⅱ.There was a significant increased in the degree of VEGF-C and VEGF-D expression in positive lymph node metastasis group(VEGF-C:24±13,VEGF-D:3.9±2.0)compared with negative group(P＜0.05).(4)There were significant positive correlations between the expression levels of Proxl and VEGF-D(r=0.62,P＜0.01),PDGF-C(r=0.91,P＜0.01)or PDGF-D(r=0.61.P＜0.01).Conclusions VEGF-A,VEGF-C and PDGF-A may promote lymphatic metastasis in epithelial ovarian carcinoma though else mechanisms other than lymphangiogenesis.VEGF-D may facilitate lymphangiogenesis and lymph node metastasis in epithelial ovarian cancer.There is no significant correlation between the expression of PDGF-B and lymphangiogenesis and lymph node metastasis.PCGF-C and PDGF-D may motivate lymphangiogenesis,but could not participate in lymph node metastasis in ovarian carcinoma.

Objective:To evaluate the hemostatic effect of self-expanding polyurethane foam in an animal model of fatal hepatic trauma and hemorrhage.Methods:The fatal liver trauma hemorrhage model with swine was established. Then the damage-controlled resuscitation was performed. Thirty minutes after injury, the experimental animals were randomly divided into the gauze packing group (GP), foam packing group (FP) and blank control group (BC). The survival time, vital signs, the bleeding volume, coagulation function and other lab indicators were recorded for 48 h. Liver histopathological examination was performed after death or execution.Results:All the three groups had severe hemorrhagic shock after modeling. The 48-h survival rate of the FP group and the GP group was significantly higher than that of the BC group (6/6 vs 4/6 vs 0/6). The average survival time of the FP group was not statistically different from that of the GP group [48 h vs (44.58±5.53) h, P>0.05], and was significantly longer than that of the BC group [48 h vs (1.64±0.17) h, P<0.01]. The bleeding volume of the FP group was significantly less than the GP group and BC group [(19.2±7.3) g/kg vs (41.3±8.6) g/kg, (51.5±7.3) g/kg, both P<0.01]. Compared with the GP group and the BC group, the cardiac output of the FP group was significantly improved [(5.00±0.53) L/min vs (4.13±0.41) L/min, (2.38±0.48) L/min, both P<0.05]. The coagulation function, liver and kidney function and blood lactate level of the FP group and the GP group were better than those of the BC group; the intra-abdominal pressure of the FP group was significantly higher than that in the GP group [(18.83±3.25) cmH 2O vs (3.83±1.47) cmH 2O, P<0.05]. There was no abnormal increased in intra-abdominal pressure in the BC group. According to the histopathology examination, there was no obvious secondary damage in the FP group. Conclusions:The application of self-expanding polyurethane foam for intraperitoneal packing to stop bleeding can effectively reduce the amount of bleeding in the fatal liver trauma hemorrhage model, effectively maintain vital signs, and improve the short-term survival rate.

Objective To study the efficacy and safety of estradiol and drospirenone tablets (Angeliq)in treatment of menopausal symptoms among postmenopausal Chinese healthy women.Methods Total 244 postmenopausal Chinese healthy women who had moderate to severe hot flushes were randomly assigned for 16 weeks in this randomized multi-center double-blind placebo-controlled study.During the trial.the follow-up visits were conducted at week 4,8,12,16 of treatment and 2 weeks after treatment respectively.Height,weight,vital signs,hot flushes,other relevant menopausal symptoms and vaginal bleeding were observed in each follow-up visit,while the clinical global impression scale Was assessed at 16 weeks as well.Results It showed that hot flushes were reduced significantly more in observation group than that in placebo group ( P＜0.01 ), although both treatments were effective. The absolute values of mean severity index of total hot flushes decreased by - 0. 6± 0. 5 in observation group and - 0. 4 ± 0. 4 in placebo group from baseline respectively, which reached significant difference ( P ＜ 0. 05 ). However, the absolute values of mean severity index of moderate to severe hot flushes decreased by - 0. 6± 0. 8 in observation group and -0. 3± 0.6 in placebo group from baseline respectively, which had no significant difference (P ＞ 0. 05 ).After 16 weeks treatment, it also showed that estradiol and drospirenone had significant better efficacy than placebo on moderate to severe sweating, vaginal dryness and clinical global impression scale (P ＜0. 01 ).During the trial, blood pressure in observation group was stable. The rate of vaginal bleeding in observation group was higher than that in the placebo group, especially during the week 4 to week 8 when 48. 9% (87/178) in observation group and 10. 7% (6/56) in placebo group of patients bled. Although the cumulative amenorrhea rate of observation group was lower than that of placebo group in each cycle (28 days), it increased gradually along with duration of the treatment. The commonest adverse event in observation group was breast tenderness which accounted for 12.0% (22/183 ). The level of serum potassium was in the normal range in observation group mostly. Meanwhile, the other adverse events rate was low. Serious adverse events reported in this trial were assessed as not study drug related or as unlikely study drug related. Conclusion Estradiol and drospirenone tablets which could effectively alleviate menopausal symptoms in postmeuopausal Chinese healthy women is a novel hormone replacement therapy regimen with high safety and efficacy.