Objective:To develop HMGA1-silencing SMMC-7721 cell and investigate its impacts on apoptosis and cell cycle in tumor cells.Methods:Constructing HMGA1-silencing SMMC-7721 cell through G418 selection of RNAi plasmid transfected cells;surveying the apoptosis,prliferation and cell cycle of the tumor cells by FACS and MTT.Results:Stable HMGA1-silencing cells were obtained successfully.The apoptosis percentage in RNAi group(29.46?3.04%) was significantly higher than that in the negative control (1.96?0.76%) or control (2.04?0.70%)within each P

Objective:To establish a mouse fibroblastic cell line stably transfected with HMGA1 gene,and to use the cell line to investigate the role of HMGA1 in tumor development and progression.Methods:Eukaryotic expression vector pcDNA3.0-HMGA1 was transfected into NIH3T3 by lipofectamine-mediation.Stable transfectants were selected by G418.The expression of extraneous HMGA1 gene was analyzed by RT-PCR and DNA sequencing.Cell growth was measured through MTT and the cell cycle by FCM.Soft agar colony formation was employed to analyze the ability of anchorage-independent growth.The expression of the immune inhibition factors of VEGF and FasL mRNA was detected by RT-PCR.Results:NIH3T3 cell lines stably expressing HMGA1 gene were successfully established.Comparing with controls,the HMGA1 gene-transfected cells grew faster,acquired the ability of anchorage-independent growth and expressed the immune inhibition factors of VEGF and FasL mRNA.Conclusion:Extraneous expression of HMGA1 gene can induce malignant transformation of NIH3T3 and modulate mRNA expression of immune inhibition factors.HMGA1 gene plays a key role in tumor development,progression and immune escape.

Objective To investigate the methods and effects of posterior fixation and fusion in treatment of complete thoracolumbar fracture and dislocation.Methods The study enrolled 8 cases of complete thoracolumbar fracture and dislocation treated by posterior fixation and fusion with pedicle screwrod system between January 2006 and December 2012.There were 7 males and 1 female,at mean age of 31.9 years (range,19-49 years).Mean time interval between injury and surgery was 8.1 days (range,4-12 days).Fracture-dislocation classification was AO type C,Denis three-column injury,and Meyerding grade V.According to American Spinal Injury Association (ASIA),there 5 cases at grades A,1 at grade B,1 at grade C 1 at grade E.Fracture-dislocation segments included T5-T6 in 1 case,T12-L1 in 3,L1-L2 in 2,L3-L4 in 1 and L4-L5 in 1.Results Mean duration of surgery was 220.6 minutes (range,135-335 minutes) and mean intraoperative blood loss was 1 150 ml (range,500-2 400 ml).Seven cases sustained dural laceration during the operation,which were sutured or covered with autologous fat grafts,but 3 of them were subjected to cerebrospinal fluid leakage and healed after conservative therapy.Anatomic reduction was achieved in 6 cases,partial reduction in 1 and non-reduction in 1.Mean Cobb angle improved from 29.3 ° (range,8 °-51 °) preoperatively to 1.9 ° (range,-5°-10 °) postoperatively.After a mean follow-up of 39.3 months (range,2-76 months),2 cases were recovered from preoperative ASIA grade A and B to C respectively and 6 cases (4 A,1 C,1 E) revealed no significant improvement.There was no implant loosening or breakage.One case was died of lung-related complications at postoperative 4 years.One case sustained lumbar deep infection at postoperative 3 weeks and managed by debridement,irrigation,drainage and implant retention.Conclusion Posterior fixation and fusion is the general treatment principle for complete thoracolumbar fracture and dislocation,but the degree of reduction depends on severity of the injured spinal cord.

OBJECTIVE:To prepare Puerarin polymeric micelles and establish a method to determine its entrapment efficiency. METHODS:Puerarin polymeric micelles were prepared by film dispersion method. The polymeric micelles and free drug were sepa-rated by centrifugal-millipore filter filtration method. The entrapment efficiency of puerarin polymeric micelles was determined by HPLC. Diamonsil C18(2)column was used with 1% citric acid solution-methanol(65∶35)at the flow rate of 1 ml/min. The detec-tion wavelength was set at 250 nm,and column temperature was room temperature. RESULTS:The prepared polymeric micelles were spherical and spherical-like in shape with a mean particle size of 54.12 nm,polydispersity index of 0.122,Zeta potential of -13.60 mV;the linear range of puerarin was 2-10μg/ml(R2=0.999 4)with average recovery rate of 99.2%(RSD=0.9%,n=3). The re-covery rate of free drug was 95.3%(RSD=1.7%,n=3). The mean entrapment efficiency and drug-loading amount of puerarin were(35.5±2.12)% and(0.3±0.07)%,respectively(n=3). CONCLUSIONS:Film dispersion method is suitable for the prepara-tion of Puerarin polymeric micelles. Established method is convenient,accurate and reliable for the content and entrapment efficien-cy determination of Puerarin polymeric micelles.

Objective To investigate the role of opioid receptors in the protective effects of isoflurane-induced delayed preconditioning against myocardial ischemia-reperfusion (I/R) injury in rabbits. Methods Forty male New Zealand white rabbits weighing 2.0-2.5 kg were randomly assigned into 4 groups ( n = 10 each) : group I sham operation (S); group II I/R; group Ⅲ isoflurane + I/R (Iso) and group IV Iso + naloxone + I/R (Nal). Myocardial I/R was induced by 40 min occlusion of left anterior descending branch (LAD) of coronary artery followed by 120 min reperfusion. In group Ⅲ (Iso) 2% isoflurane in 100% O2 was inhaled for 2 h and I/R was produced 24 h later. In group IV (Nal) naloxone 6 mg/kg was given iv 10 min before 2 h of 2% isoflurane inhalation and I/R was produced 24 h later. At the end of 120 min reperfusion, infarct size (IS) and area at risk (AAR) were determined by Evan's blue and TTC staining. Myocardial ultrastructure was examined by electron microscopy. The phosphorylated p38MAPK protein expression in myocardium was determined by Western blot. Results The IS was significantly smaller in group Iso ( Ⅲ ) ( 19.7% ± 2.8%) than in I/R group ( II ) (37.8% ±1.7%) (P<0.05). The phosphorylated p38MAPK protein expression in myocardium was significantly lower in group Iso than in group I/R. Microscopic examination showed less myocardial damage in Iso group than in group I/R. The protective effects of delayed preconditioning by isoflurane was prevented by naloxone pretreatment. ConclusionOpioid receptors may be involved in the protective effects of delayed preconditioning by isoflurane against myocardial I/R injury.

OBJECTIVETo investigate the modulatory effects of morphine on the susceptibility to pentylenetetrazole-induced seizures, and the involvement of endogenous histamine in this process.

METHODSBoth the wild-type (WT) mice and histidine decarboxylase (a key enzyme for histamine biosynthesis) deficient (HDC-KO) mice were subcutaneously injected with different doses of morphine, and 1 hour later the pentylenetetrazole solution (1.5 %) was infused into the tail vein at a constant rate of 0.3 ml/min. The minimal dose of pentylenetetrazole (mg/kg) needed to induce myoclonic jerks and clonus convulsion was recorded as the thresholds of seizures.

RESULTIn WT mice, morphine dose-dependently decreased the thresholds of both myoclonic jerks and clonus convulsion. In HDC-KO mice, morphine at 10 mg/kg only significantly decreased the threshold of myoclonic jerks from (38.6 +/-2.9)mg/kg to (32.5 +/-0.7)mg/kg, but had no significant effect on the threshold of clonus convulsion [from (51.8 +/-2.1)mg/kg to (47.6 +/-1.2)mg/kg]. In addition, the value of decreased myoclonic jerks (15.8 +/-1.4)% and clonus convulsion (8.3 +/-0.9)% thresholds were much lower in HDC-KO mice than in WT mice [(26.1 +/-2.5)% and (20.8 +/-2.4)%, respectively].

CONCLUSIONMorphine can decrease the thresholds of pentylenetetrazole in induction of seizure, and the endogenous histamine may be involved in this process.

Animals ; Disease Susceptibility ; chemically induced ; metabolism ; physiopathology ; Dose-Response Relationship, Drug ; Histamine ; metabolism ; physiology ; Histidine Decarboxylase ; genetics ; metabolism ; Male ; Mice ; Mice, Knockout ; Morphine ; pharmacology ; Myoclonus ; chemically induced ; metabolism ; physiopathology ; Narcotics ; pharmacology ; Pentylenetetrazole ; Random Allocation ; Seizures ; chemically induced ; genetics ; physiopathology ; Sensory Thresholds ; drug effects

Objective To prepare specific mouse monoclonal antibodies against Homo sapiens dynein,axonemal, heavy chain 2 (DNAH2). Methods Firstly, recombinant plasmid encoding His tagged immunogen, targeting N-terminal sequence of DNAH2 protein (1-300 aa), in E. coli was constructed. IPTG was used to induce the expression of His-immunogen, which was then purified and immunized in BALB/c mice. Hybridoma cells were obtained through the fusion between myeloma cells and splenocytes isolated from BALB/c mice. Finally, ELISA and Western blot assays were performed to screen the positive hybridoma. Results IPTG was used efficiently to induce the expression of DNAH2 immunogen in E. coli. DNAH2 protein bands were detected in screened positive hybridoma. Conclusion Mouse monoclonal anti-DNAH2 antibody is prepared successfully.

0.05).There was significant difference in the 3-year survival rate between A and C group. Conclusions The 3-year survival rate was dramatically increased with combined therapy mainly by cisplatin, the dose of 60～80mg is tolerant for the elderly aged above seventy years, and perioperation complications can be cured.

Objective To investigate the effect of calcium antagonist verapamil on the function of rat?- cells and tolbutamide (D860)-induced insulin release.Methods Insulin released from isolated islets were measured in control,verapamil,D860,and verapamil+D860 groups.Furthermore,intravenous glucose tolerance test (IVGTT) was conducted in acute experiments treated with verapamil and D860 respectively to assess?-cell function in rats with the same allocation as in vitro.Another IVGTT was performed in the end of 4 weeks' treatment.The insulin contents in pancreas were assayed and pancreas islets morphology were observed with immunohistochemistry.Results Verapamil could inhibit insulin release from isolated islets.Verapamil group was [(1.244?0.082)ng?ml~(-1)?islet~(-1)]and control group (2.623?0.226) ng?ml~(-1)?islet~(-1)(P0.05).Also,similar results were obtained in normal rats during acute experiments and verapamil reduce the hypoglycemic effect promoted by D860. However,above results were not observed in the end of 4 weeks experiments,and no difference for insulin content and morphological change in islets was found among four groups.Conclusion Treatment of verapamil chronically does not impair islet function and interfere with the hypoglycemic effect of D860 in rats .

Objective To observe the MRI features of traumatic brainin jury to corpus callosum and provide valuable diagnostic information for clinical application. Methods Clinical and radiological data were retrospectively analyzed in 20 MRI-diagnosed patients as having traunmtic brain mjury to corpus callosum. Results Traumatic injury to corpus callosum was rather rare in brain trauma.Most traumatic injury to corpus callosum was non-hemorrhagic and its lesion mainly laid at its body or splenium,sometimes at genu,but rarely at rostrum.Traumatic injury to corpus callosum,being one of the common manifestations in diffuse axonal injury(DAI), could be demonstrated slight hyperintense signal on TI WI and T2WI in early phase.The injury site shrank and liquefaction necrosis appeared in the lesion at the late-mid-injured period with softened focus or gliai scar.MRI showed hyperintense T1 and T2 signal, similar to the signal of the cerebrospinal fluid and demonstrated ventriculomegaly in the corresponding parts.DWI showed hyperintense signal in the lesion in acute and subacutc stages:with time prolonged,isointense signal of the normal brain tissue was found and signal of the cerebrospinal fluid appeared in the present of softened focus.Conclusion Attention should be paid to the traumatic injury to corpus callosum,an indicator for severe trauma or disease.MRI,a sensitive and multi-directional imaging,is superior in detecting non-hemorrhagic injury to the corpus callosum.