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 compare the immediate therapeutic effects between endovenous laser treatment and conventional surgery for lower extremity varicosity.Methods Data of 350 limbs of 275 patients treated by endovenous laser treatment (endovenous laser treatment group) and 310 limbs of 210 patients by traditional surgery (traditional surgery group) were analyzed and compared in terms of operation time,number of the incision,postoperative pain,complications,postoperative hospitalized duration and 1-year recurrence rate.Results Endovenous laser treatment group had shorter operation time,fewer incisions,less postoperative pain and shorter postoperative hospitalized duration than traditional surgery group (P0.05).Conclusion As a safe and effective new treatment for lower extremity varicosity with minimal invasiveness and no left scars,endovenous laser treatment has the potential to replace conventional surgery and extend the surgical indications for lower extremity varicosity treatment.

Objective:The different subtypes of voltagE-gated sodium channels (VGSCs) are known to correlate with the migration of many malignant cancers. This study was to investigate the significance of functional expression of SCN5A/Nav1.5 in human ovarian cancer and its effects on migration capability of ovarian cancer cells in vitro. Methods: Sodium indicator SBFI and immunofluorescence method were used to detect the distribution of intracellular Na+. Real-time PCR, Western blotting, and immunohistochemistry were used to detect the mRNA and protein expression of SCN5A/Nav1.5. The effect of specific voltagE-gated sodium channels inhibitor tetrodotoxin (TTX) on cell viability was measured by CCK-8 kit. The migration and invasion of ovarian cancer cell lines SKOV-3 were tested by Transwell chamber assay. Results:SCN5A/Nav1.5 were over-expressed in ovarian cancer cell lines SKOV-3 and ovarian cancer specimens at mRNA and protein levels. TTX 30 μmol/L inhibited the intracellular Na+ concentration by (41.51±0.41)%. TTX also suppressed the invasion and migration capacities of SKOV-3 cells by (33.80±1.6)% and (43.60±2.9)%, respectively. The difference was significant (P<0.05). Conclusion:SCN5A/Nav1.5 is involved in the metastasis progression of ovarian cancer in vitro and plays an important role in the initiation and progression of ovarian cancer. It may become a target for ovarian cancer therapy.

Objective To investigate the irradiating effect of low intensive microwave(LIM) on pathological process of blood vessel restenosis(RS) and assess the probability of LIM irradiation to prevent was used RS.(Methods Forty-four) male healthy New Zealand rabbits were randomly divided into 2 groups.Fogarty catheter traumatize to the tunica intima of iliac artery so as to establish RS models.Two thousand four hundred and fifty MHz microwave with different power of 2,5 and 10 mW/cm~2 was used,locally to irradiate EIA in irradiating group(1 h/d).Specimens were obtained at different time of 3,7,14 and 28 d after operation.Morphological changes of tissues were observed with HE and EF staining and the area of tunica intima,tunica media and the rate of cavity stenosis were analyzed with image analysis system;apoptosis was detected with TUNEL;phenotype and microstructure of VSMC were observed with TEM.Results After microwave irradiating,inflammatory reaction in early period was suppressed,mural thrombus decreased,the proliferation and migration of VSMC depressed,the area of tunica intima and the rate of cavity stenosis obviously reduced comparing with the control group(P0.05).At other different time,however,the rate of apoptosis cells in irradiating groups obviously increased than that of the control group(P

For treatment of pediatric inguinal hernia, we fabricated a device, i.e. so called "filling type pediatric hernia sac", which treats the problem from the abdominal cavity, through the abdominal and is a self-adaptive closer, using synthetic material. The device includes filling rack, self-adaptive umbrella support bar, bottom piece, outside pulling line and device fixing lines. The filling rack is composed of 2 concentric circles of 3.0 cm diameter with peripherally fixed together and can be pulled into the shapes of a ball or an olive. The supporting bar is structured of 3 pieces with 0.5 cm wide, 4.0 cm long, cross-fixed on top of the filling rack. The bottom piece is in a circular structure with a diameter of 3.0 cm, and it is connected to the filling rack bottom. Adjust positioning stay outside the fixed on the top of the device are connected at one end, and the other end free through filling the top frame connected with the bottom slice of central fixation. By using this device, we treated 37 pediatric inguinal hernia cases with 38 side-inguinal hernia successfully. The mean duration of post-operation follow-ups was 14.6 ± 5.89 months, without hernia recurrence, obvious scar and hard sections of inguinal region. This device could provide a convenient, safe and effective plugging technology for children's pediatric hernia.
Child ; Hernia ; therapy ; Herniorrhaphy ; instrumentation ; Humans

Objective To investigate the effects of early rehabilitation therapy in mechanically ventilated ICU in patients.Methods A randomized controlled trial was carried out.Sixty mechanically ventilated patients,with tracheal intubation or tracheostomy more than 48 h and less than 72 h,were admitted to the intensive care unit (ICU) of the Affiliated Hospital of Medical College Qingdao University from May 2010 to May 2012.These patients were randomly (random number) divided into two groups,the rehabilitation group and the control group,30 patients in each group.In rehabilitation group,rehabilitation therapy was taken twice daily,and the training time and intensity was adjusted according to the condition of the patients.Early rehabilitation therapy included head up actively,transfer from the supine to sit,out of bed,transfer to a chair,standing bedside bed and walking bedside bed.The patient' s body mass index,days to first out of bed,duration of mechanical ventilation,ICU stay,APACHE Ⅱ score,highest FiO2,lowest PaO2/FiO2 and hospital mortality of patients were compared between rehabilitation group and control group.The differences between the groups were compared using t test.Results There was no significant difference in body mass index,APACHE Ⅱ score,highest FiO2,lowest PaO2/FiO2 and hospital mortality between rehabilitation group and control group (P ＞ 0.05).Patients in the rehabilitation group had shorter first out of bed time [(3.8 ± 1.2) d vs.(14.9 ±4.7) d; P =0.00],duration of mechanical ventilation [(5.6±2.1) dvs.(12.7±4.1) d; P=0.005] andICUstay [(7.3±2.8) dvs.(15.2±4.5) d;P =0.01] compared with control group.Conclusions Early rehabilitation therapy was safe and effective in improving the outcomes of mechanical ventilation patient.

Voltage-gated sodium channels (VGSCs) are known to be involved in the initiation and progression of many malignancies, and the different subtypes of VGSCs play important roles in the metastasis cascade of many tumors. This study investigated the functional expression of Nav1.5 and its effect on invasion behavior of human breast cancer cell line MDA-MB-231. The mRNA and protein expression of Nav1.5 was detected by real time PCR, Western Blot and immunofluorescence. The effects of Nav1.5 on cell proliferation, migration and invasion were respectively assessed by MTT and Transwell. The effects of Nav1.5 on the secretion of matrix metalloproteases (MMPs) by MDA-MB-231 were analyzed by RT-PCR. The over-expressed Nav1.5 was present on the membrane of MDA-MB-231 cells. The invasion ability in vitro and the MMP-9 mRNA expression were respectively decreased to (47.82+/-0.53)% and (43.97+/-0.64)% (P<0.05) respectively in MDA-MB-231 cells treated with VGSCs specific inhibitor tetrodotoxin (TTX) by blocking Nav1.5 activity. It was concluded that Nav1.5 functional expression potentiated the invasive behavior of human breast cancer cell line MDA-MB-231 by increasing the secretion of MMP-9.

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.

BACKGROUND:Hip arthroplasty and internal fixation are presently effective therapeutic methods in treatment of femoral neck fracture in the elderly. However, which method can reduce the incidence of postoperative complications remains controversial.
OBJECTIVE:To systematical y review the reoperation, postoperative complications and 1-year and 2-year mortality after hip arthroplasty and internal fixation in the elderly with femoral neck fracture.
METHODS:Pubmed/Medline, EMBASE, and Cochrane CENTRAL databases were retrieved by computer for articles published before May 2013. Systematic review on randomized control ed trials of hip arthroplasty versus internal fixation for femoral neck fractures in the elderly was conducted using the Cochrane Col aboration’s RevMan 5.2 software. Outcome measures included reoperation, main complications related to the surgery and mortality.
RESULTS AND CONCLUSION:Six published randomized control ed trials of nine literatures containing 1 496 cases were involved in this review. Meta-analysis results indicated that reoperation rate was greater in the internal fixation group within and more than 2 years after the surgery compared with the hip arthroplasty group (P<0.000 01). Compared with hip arthroplasty group, internal fixation significantly increased the main complications related to the surgery [OR=8.79, 95%CI(6.50-11.88), P<0.000 01]. No significant difference in 1-year and 2-year mortality after surgery was detected between the internal fixation and hip arthroplasty groups [OR=0.85, 95%CI(0.66-1.11), P=0.23;OR=0.88, 95%CI(0.70-1.10), P=0.27]. These data suggested that the long-term reoperation rate and incidence of main complications were obviously higher in internal fixation compared with hip arthroplasty for femoral neck fracture in the elderly, and no significant difference in 1-year and 2-year mortality after the surgery was detectable between the two methods. Clinical recommended hip arthroplasty in the repair of femoral neck fracture in the elderly.

Objective To observe the effects of TLR9 on the nude mouse transplanted tumor growth of human pancreanc cancer and its drug resistance.Methods The nude mouse transplated tumor of human pancreatic cancer PANC-1 was established and randomly divided into 6 groups for conducting the experiment:sterile normal saline group,TLR9 agonist,TLR9 inhibitor group,gemcitabine group,TLR9 inhibitor plus gemcitabine Bin group,TLR9 agonist plus gemcitabine.The tumor size and growth situation were recorded by the vernier caliper.The immunohistochemical method was used to detect tumor TLR9 receptor expression.The tumor growth,metastasis and paracancerous nssue invasion situation were observed by the magnetic resonance imaging (MRI).Results The volume and growth speed of resected tumor mass in the gemcitabine group,TLR9 agonist + gemcitabine group,TLR9 inhibitor plus gemcitabine group was significantly smaller than those in other groups (P＜0.05),which in the TLR9 agonist + gemcitabine group were significantly greater than those in the TLR9 inhibitor plus gemcitabine group and gemcitabine group (P＜0.05),the difference between the TLR9 inhibitor plus gemcitabine group and gemcitabine group had statistical significance (P＜0.05),while the difference among the TLR9 agonist group,TLR9 inhibitor group and normal saline group had no stastistical significance (P＞0.05).The tumor in mice at 7 weeks after planting showed oval shape with clear boundary by MRI observation,no obvious metastais and paracancerous invasion were seen in paracancerous nssues no statistically significant,5 weeks,6 weeks after planting,seven weeks mice observed in MRI,the tumor into an,state clearly that the transfer of the surrounding tissue,no significant vascular invasion,heart,liver,kidney disease.The TLR9 expression on the surface of tumor tissue was detected and identified.Conclusion Pancreatic cancer nude mouse transplated tumor has definitely positive expression of TLR9,TLR9 activation can significantly decrease the sensitivity of pancreatic cancer to chemotherapy,increases the drug resistance of tumor,on contrary promotes the tumor growth.