Objective To discuss the influences of programmed death-1 (PD-1) factor on osteosarcoma cells MG-63.Methods Osteosarcoma stem cells were sovted and identified through osteosarcoma cell strain MG-63 cells.The influence of PD-1 signal on T cells proliferation were detected by MTT. The expression of PD-1 mRNA was detected by RT-PCR.Results Cancer cells had an obvious proliferation within one week, which showed the strong ability of proliferation and aggressivity.The formation of tumor cells spheres depended on the support of serum nutrition.The number of MG-63 cells proliferation in serum culture medium was significantly higher than the osteosarcoma cells spheres in serum-free suspension culture ( P<0.05 ) .Pluripotent stem cell marks that the expression of CD133 in cancer cell sphere was significantly higher than that of MG-63(P<0.05).RT-PCR results showed the PD-1 expression level of cancer cell sphere and MG-63 were increased significantly.Conclusion MG-63 cell line has the character istics of osteosarcoma stem cells.MG-63 cell line can express the corresponding cell markers.The expression of PD-1 also increase significantly which can reduce immune function of patients and is closely related with the occurrence and development of tumors.

ObjectiveTo compare and evaluate the effectiveness of Airtraq laryngoscope combined with a Bougie and Airtraq laryngoscope alone for tracheal intubation in simulated difficult airway.Methods Four anesthetists and 4 clinical physicians of standardized training were enrolled in the study.The participants intubated the trachea of the ALS simulator manikin in 5 tongue edema scenarios simulating modified Cormack-Lehane grade 1,2a,2b,3,and 4 views and 1 cervical immobilization scenario.Results No significant difference in the rate of successful intubation was detected between two techniques(P＞ 0.05 ).In Cormack-Lehane grade 1,2a views,the duration of successful intubation in Airtraq laryngoscope alone [ ( 14.3 ± 1.3),( 17.1 ± 2.9) s] was shorter than that in Airtraq laryngoscope combined with a Bougie [ (26.6 ± 3.8),(36.4 ± 3.6) s ] with significant difference (P ＜ 0.01 ).Cormack-Lehane grade 2b,3,4 views,the duration of successful intubation in Airtraq laryngoscope alone[ (74.5 ± 6.5 ),(116.3 ± 9.8),(53.0 ± 6.1 )s] was longer than that in Airtraq laryngoscope combined with a Bougie [ (35.4 ± 4.3 ),(52.3 ± 5.0),(40.4 ± 3.8 ) s ] with significant difference (P ＜ 0.05).ConclusionAirtraq laryngoscope combined with a Bougie can be quickly intubated in simulated difficult airway compared with Airtraq laryngoscope alone.

Objective To construct a lentiviral expression vector of murine RelB for RNAi,and then to effectively silence the RelB gene expression of murine bone marrow derived dendritic cells for constructing the bone marrow tolerogenic dendritic cell,in order to provide an experimental foundation for a novel clinical therapy of autoimmune disease.Methods RelB shRNA sequence of mouse was designed by on-line designer software on Corp.Invitrogen,after synthesis and annealing,double strand oligonucleotides(dsoligoes)were cloned into the pENTRTM/U6 plasmid,then after sequencing,a positive clone was further subcloned into pLenti6/BLOCK-iTTM-DEST vector.It was then transformed into stb13 competent cell,and after sequencing,293FT cell line was transfected by above positive recombined plasmid and lentiviral packing materials.It was incubated for 48 to 72 h in a 37℃,5% CO2 incubator.Culture supernatant was harvested and stored at-80℃,then the virus titer was determined by serial dilution assay.Results It was showed from sequencing figures that all the pENTRTM/U6-RelB-shRNA plasmids were positive clone vector,and this positive recombinant vector was recombined with pLenti6/BLOCK-iTTM/U6-DEST vector.It was then transformed into stb13 competent cell and screen positive clone by ampicillin,and resequenced by the use of primer forward U6 primer.The results also showed that recombinant lentiviral vector was positive clone.Viral particle was packaged with other packaging material mediated by lipofectamine 2000 in 293FT cell line.Cultural supernatant was collected and stored at-80℃,and lentiviral particle titer was determined by serial dilution assay with 6?105/transduced unit.Conclusion Lentiviral shRNA expression vector of murine RelB gene for RNAi was successfully constructed.It might be a rational procedure to make tolerogenic DC,and then to develop a DC vaccine and DC-based immunotherapy for autoimmune diseases.

Object To study the effect of geniposide in the prophylaxis of cholesterol calculus formation in gall ducts of Syria Golden hamster Methods Experimental hamster models with cholesterol calculus was prepared by feeding lithogenous diet with high content of fat and protein for 30 d with simultaneous administration of geniposide at doses of 50 and 100 mg/kg/d ig or ursodeoxycholic acid as positive control at the dose of 80 mg/kg/d After the last dosage, the hamsters were sacrificed and their gallstone formation rate were determined by examining the smear of gallbladder content and their lithogenous index (LI) were calculated from the assay of lipoid substances in bile such as cholesterol (Ch), bile acids (Bs) and phospholipid (Pl) Results The gallstone formation rate in the blank model group, geniposide 50 and 100 mg/kg were 100%, 40% (P

Humans ; MicroRNAs ; Occupational Medicine

BRCA1 Protein ; genetics ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Designer Drugs ; therapeutic use ; Humans ; Lung Neoplasms ; drug therapy ; Precision Medicine ; methods ; Tubulin ; genetics ; Xeroderma Pigmentosum Group D Protein ; genetics

Objective To explore the methods and clinical effects of dimple plasty with syringe needle guiding.Methods From February 2008 to May 2012,36 dimple plasties in 21 patients were performed with using a syringe needle guiding the absorbable suture,including 15 cases in bilateral side and 6 cases in unilateral side.There were 4 cases in unilaterally right side and 2 cases in unilaterally left side.Two patients with inborn dimple in unilateral side received the dimple plasty in the other side for facial symmetry.Results 34 dimples in 21 patients were satisfied with the results.Two dimples were re-operated after three months because of disappearance.There were scleromas in 2 cases,which was disappeared by hot compress in three months.There were no hemotoma and infection in 21 patients.Conclusions The procedure of dimple plasty under syringe neddle guiding is safe,reusable and effective.It is a minimally invasive surgery and the patients are satisfied with the dynamic results after operation.

Objective To develop an all-in-one multifunctional thoracolumbar fixation rescue vest for land and water rescue of thoracolumbar injury and investigate its application effect.Methods According to the thoracolumbar physiological curve,type Ⅰ thoracolumbar fixation rescue vest with keel and inflatable airbag and type Ⅱ thoracolumbar fixation rescue vest with additional lifesaving balloon and inflatable cylinder were developed using thermoplastic polyurethane (TPU)-nylon composites and polycarbonate (PC) as the main materials.Clinical application of type Ⅰ thoracolumbar fixation rescue vest in 532 cases of thoracolumbar injury was discussed.Type Ⅱ thoracolumbar fixation rescue vest were tested at marine rescue and related parameters measured were automatic inflation time,buoyancy force,floating time,floating condition and victims' face orientation.Clinical outcome was quantified by the MacNab standard,and VAS for pain was recorded.Results According to the MacNab standard,excellent outcome was achieved for all cases.VAS improved from 8.2 points to 2 points after the bracing was applied,showing 100％ improvements.The brace into the water showed automatic inflation time of 3-5 seconds and maximum buoyancy force of 100 kg,and ensured a 90 kg dummy of floating over 96 hours.At marine rescue,the wounded in braced condition showed face upward with 24-hour buoyancy loss ≤5％ and freedom of movement.Conclusions The multifunctional thoracolumbar fixation rescue vest provides dual immobilization and ensures marine rescue for its automatic inflation device.This invention provides the feasibility to remove,transport and evacuate the thoracolumbar fracture patients in cabin.

Objective To explore the effects of ligustrazine on secretion of nitric oxide(NO) in ventricular tissues of rats under physiologi-cal condition.Methods Twenty-four normal wistar rats were divided into three groups,eight each group.The left and right ventricular tissues of each rats were divided into control group and 2 ligustrazine groups and the final concentrations of ligustrazine were 15 mg/L and 150 mg/L respectively.The ventricular tissues that had been cutting into small pieces were incubated in 37 ℃ thermostatic water bath for 4 hours with Krebs solution as supernatant.After incubation of the ventricular tissues,the spectrophotometric method was used to detect the content of NO_2~-/NO_3~-in the supernatant.Results Compared with the content of NO in control group,it was significantly decreased in the 150 mg/L ligustrazine group(P

Objective To observe the influence of serum containing Rhizoma Chuanxiong on the proliferation and differentiation of mouse embryos stem cells(ESCs).Methods Mouse ESCs were co-cultured with serum containing Rhizoma Chuanxiong.The proliferation of ESCs was detected by CCK-8 method.The expression level of specific gene beta-myosin heavy chain (β-MHC) in cardiac myoblasts was detected by reverse transcription-polymerase chain reaction (RT-PCR).Results Compared with the blank rat serum group and the blank fetal bovine serum group,the differences of activities of ESCs in serum containing Rhizoma Chuanxiong group were in significant(P ＞0.05) and the expression level of specific gene β-MHC in cardiac myoblasts was increased (P ＜ 0.05).Conclusion Serum containing Rhizoma Chuanxiong can promote the differentiation of ESCs into cardiac myoblasts.