Objective To investigate whether RNA interference in mediated Bcl-2 gene silence can decrease the gallbladder carcinoma xenograft formation rate and inhibit growth of xenograft in nude mice.Methods In the xenograft formation ability group,the Bcl-2-siRNAs were transfected into the gallbladder carcinoma cell.Transfected gallbladder carcinoma and gallbladder carcinoma were injected into nude mice hypodermically to form a gallbladder carcinoma tumor.The formation rate and volume of the tumors were measured and the tumor growth rates were calculated.In the therapy group,the animal models of human gallbladder carcinoma in nude mouse body were constructed.The nude mice were randomly divided into pSilencerTM-EGFP sh515 group,pSilencerTM-EGFP shCon group,and normal control group.After the formation of tumor knot,siRNA expressing plasmids pSilencerTM-EGFP sh515 and pSilencerTM-EGFP shCon were injected into the knots of pSilencerTM-EGFP sh515 group and pSilencerTM-EGFP shCon group.The tumor volume was measured and tumor growth rate was calculated.All mice were sacrificed after 6 weeks.The distribution of Bcl-2 in the tumors was detected by immunohistochemistry.Results The nude mouse model of human gallbladder cancer xenograft tumor was constructed successfully.Xenograft formation ability:The xenograft formation rate of gallbladder carcinoma and Bcl-2 siRNA transfected gallbladder carcinoma was 100% and 60%,respectively;the average tumor volume in gallbladder carcinoma group and transfected gallbladder carcinoma group was(1 914.6?125.0)mm3 and(629.7?78.9)mm3;the growth rate was 45.58% and 14.99%,and the average tumor weight was(2.24?0.33)g and(0.77?0.12)g,respectively.The tumor volume,weight and growth rate in the transfected gallbladder carcinoma group decreased significantly when compared with those in control group(P

Objective To investigate the related mechanism of ligamentum flavum (LF) hypertrophy in diabetic pa-tients with lumbar spinal canal stenosis ( LSCS ) .Methods Twenty-four diabetes mellitus patients [ DM (+) ] and twenty normoglycemic patients [ DM (-) ] with LSCS were enrolled in this study .Sorbitol in LF was analyzed using D-Sorbitol/Xylitol test kit .The thickness of LF was measured by CT .The structure of LF was observed after HE and Masson's trichrome staining .The cell cycle and proliferation of fibroblastic cell NIH 3T3 line cultured in high glucose were analyzed .Sorbitol of NIH3T3 was detected under different backgrounds in vitro, normal glucose , high glucose and high glucose burdened with aldose reductase inhibitor ( ARI) , Epalrestat .The expression of inflammatory factors was detected by qPCR and Western blot under above different backgrounds .Results LF of diabetic patients exhibi-ted significantly higher level of sorbitol and pro-inflammatory cytokines , TGF-βand of CD68-positive staining than that of the normoglycemic subjects ( P<0.01 ) .The diabetic LF was significantly thicker than that of the controls , and showed evidence of degeneration .The high glucose-cultured fibroblasts exhibited significantly higher levels of sorbitol , pro-inflammatory factors , and TGF-βcompared to the low glucose-cultured cells , and these levels were dose-dependently reduced by treatment with the aldose reductase inhibitor (P<0.05).Conclusions Sorbitol level of the LF is significantly increased in the DM patients with LSCS .Increased sorbitol recruites inflammatory factors and fibrogenic-related factor TGF-βin LF of DM patients with LSCS which may contributes to the LF hypertrophy .

Objective To investigate the effect of parecoxib pretreatment on the expression of aquaporin-4 (AQP4) during focal cerebral ischemia-reperfusion (I/R) in rats.Methods One hundred and twenty-eight adult male Sprague-Dawley rats,aged 6-8 weeks,weighing 230-280 g,were randomly divided into 4 groups (n =32 each):sham operation group (S group),I/R group,parecoxib 5 mg/kg group (group P5) and parecoxib 10 mg/kg group (group P10).The rats were anesthetized with 10％ chloral hydrate 3.5 ml/kg.Focal cerebral I/R was induced by 2 h middle cerebral artery occlusion followed by reperfusion.Parecoxib 5 and 10 mg/kg were injected via the right internal jugular vein at 30 min before middle cerebral artery occlusion in groups P5 and P10,respectively.The equal volume of normal saline was injected instead of parecoxib in groups S and I/R.Neurological deficit score (NDS) was assessed at 2 and 24 h of reperfusion.The rats were then sacrificed and brains were removed for determination of infract volume (by TTC staining).Brain water content was measured by wet/dry brain weight ratio.Their brains were cut into sections which were stained with haematoxylin and eosin and examined under microscope.The expression of AQP4 in brain tissues was measured.Results Compared with S group,NDS and brain water content were significandy increased,the infarct volume was enlarged,and the expression of AQP4 in brain tissues was up-regulated in I/R group (P ＜ 0.05 or 0.01).NDS and brain water content were significantly lower,the infarct volume was smaller,and the expression of AQP4 in brain tissues was lower in groups P5 and P10 than in I/R group (P ＜ 0.05 or 0.01).Microscopic examination showed that brain injury was significantly attenuated in groups P5 and P10 as compared with I/R group.Conclusion The mechanism by which parecoxib pretreatment alleviates the focal cerebral I/R injury in rats is related to down-regulation of the expression of AQP4.

ObjectiveTo investigate the changes of serum apolipoprotein E (ApoE) in children with infectious diseases.MethodsA total of 279 pediatric patients with infectious diseases were enrolled in this study,including 65 patients with sepsis,47 patients with bacterial meningitis,67 patients with bacterial pneumonia, 47 patients with aseptic meningitis and 53 patients with mycoplasmapneumonia. TheserumApoEcollectedfromallpatientswasdetectedby immunoturbidimetric assay (IA).The septic mouse model was established by intraperitoneal injection of group B Salmonella typhimurium.Mouse serum ApoE levels were detected by IA,and the hepatic ApoE mRNA and protein expressions of mice were detected by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blot,respectively.Data in two groups were compared by independent-sample t test.ResultsSerum ApoE levels in patients with bacterial infections were increased dramatically,which was (59.8±23.5) mg/L in patients with sepsis (t=-5.118,P＜0.01),while no significant differences were found in patients with aseptic meningitis and myeoplasma pneumonia.Moreover,a high level of serum ApoE was detected in septic mouse model,while the hepatic ApoE mRNA and protein expressions of the mice were both decreased,with mRNA decreased 71％ at 3 hour (t=5.022,P＜0.01) and 73％ at 24 hour (t=4.181,P＜0.01).Conclusions Serum ApoE levels in bacterial infections increase dramatically,while its hepatic expression in septic mouse model is decreased,which indicates that the elevated serum ApoE level is not related to the changes of hepatic ApoE expression.

Objective To evaluate the effects of micro-screw on the intrusion of lower incisors and the leveling of curve of Spee in the process of orthotherapy.Methods Fifteen patients (5 males,10 females) with deep-bite were selected,aged from 11.3 to 28.8 years,with average of 17.2 years.All of them were Angle Class Ⅱ Classification 1 malocclusion and had a curve of Spee larger than 3 mm.Orthotherapy of extraction was adopted for the first premolars of two upper mandible and second premolars of two lower mandible.All these patients had micro-screws implanted at root apex area between lower lateral incisor and canine bilaterally,and when the 0.457 mm× 0.635 mm (0.018 in ×0.025 in) Ni-Tisquare wire was internalized,the strength of intrusion from micro-screws was used.Lateral cephalometric radiographs were taken before loading the micro-screw and immediately when the curve of Spee was leveled.Two linear and two angular measurements were selected for cephalometric analysis.Results The following results were noted: the lower incisors were significantly intruded for a mean of (4.10±0.98) mm; the mandibular molars were not significantly extruded for a mean of (0.91±0.69) mm; some lower incisors proclined and some lower incisors retroclined for a mean of (-0.09±4.12)°.After bite opening,the mandibular plane angle was minimally altered for a mean of (0.25±0.70)°; the ratio of lower incisor intrusioin to lower molar extrusion was 4.5: 1.0.Conclusions The combined use of micro screw with Ni-Ti wire is capable of enhancing the intrusion of lower incisors and leveling the curve of Spee,and it has minimal side effects on the posterior teeth.

Objective To discuss the method of implant for alveolar bone deficiency.Methods 37 cases of bone deficiency were chosen to use maxillary sinus augmentation,localized management of sinus floor,autologous jaw chips transplantation,alveolar bone distraction.Results All of the implants osseointegration except 1 implant loosed because of improper prosthesis.Conclusion All of the methods above provide can good ways for wider indication of implantation.

OBJECTIVE:To investigate the compatibility stability of Ulinastatin for injection with 2 commonly used solvents in the infusion pump.METHODS:After Ulinastatin for injection 500 000 U was respectively added into 0.9％ Sodium chloride injection and 5％ Glucose injection 50 mL,the appearance of the mixture at 25 ℃,37 ℃ were observed at 0,1,2,4,8,12,24 h,re spctively,pH value and the number of insoluble particles were measured.The relative percentage of ulinastatin in the mixture was determined by HPGFC.RESULTS:Under this condition,the appearance and pH value of the mixture had no significant change within 24 h;the number of particles ≥10 μm was lower than 25 particle/mL,and that ≥25 μm was lower than 3 particle/mL,which was in line with the pharmacopeia standard.The relative percentage of ulinastatin within 24 h ranged 99.45％-102.55％.CONCLUSIONS:After mixed with 0.9％ Sodium chloride injection and 5％ Glucose injection,Ulinastatin for injection keep stable within 24 h at 25 ℃,37 ℃ and can be used for continuous administration in the infusion pump.

Objective To establish the differential templates of juvenile and children in Shandong with normal occlusion, according to the group of skeletal age. Methods 212 juvenile and children at the age of 8-16 years (107 males and 105 females) with normal occlusion in Shandong natives were took cephalomatric radiographs, and divided into different groups by cervical vertebrae skeletal age. Then these cephalomatric radiographs were scanned on the computer and 14 skeletal landmarks were vectorizated. An analytical method of Ricketts and McNamara with WinCept 7.0 was used to make statistics and variance analysis among gender and every group of cervical vertebrae skeletal age,and then established the templates. Results The juvenile and children in Shandong natives with normal occlusion had different templates. Conclusion By overlapping the same skeletal age of templates by SN plane, we can see that male face outline is greater than female and male mandibular plane angle is smaller than female. By overlapping the same gender of templates, there is a developmental trend of mandibular bone to be forward and downward, and to revolve anticlockwise with age.

Objective To compare the biomechanical pull-out strength (POS) of three different fixations in upper thoracic vertebras using translaminar screws (TLS), translaminar facet screws (TLFS), and transpedicle screws (TPS), respectively. Methods Nine fresh human cadaveric cervicothoracic junction spines specimens which including T1-T3 vertebras were harvested. The vertebras specimens were scanned using dual-energy radiograph absorptiometry for bone mineral density. Both of screw insertion techniques at each vertebrae was randomized. All the screw insertions were based on direct observation and the CT scan on the pedicles. The peak of insertional torque (IT) was recorded and axial pull-out testing was performed to simulate intraoperative failure of fixation. Results The mean peak IT of the TFLS, TPS and TLS were (0.43±0.01), (0.40±0.01), (0.35±).01) N·m, respectively. There was no statistically significant difference between the TFLS and TPS, and between the TPS and TLS was same. But the TFLS generated statistically greater peak 1T in comparison with the TLS(t=-13.86, P＜0.05). The mean POS of TLFS was (771±106) N,which had no statistically significant difference in comparison with the TPS(733±65) N. And the TLS (663±86) N was same. But the TFLS generated statistically greater POS in comparison with the TLS (t=9.907, P＜0.05). The peak IT showed a strong positive correlation with POS in three screw techniques. Bone mineral density correlation with POS in all methods of fixation. Conclusion It was not a significant difference to compare POS of TLS and TLFS to that of TPS respectively. TLS and TLFS appear to be a biomechanically sound alternative in the upper thoracic spine, and appear to be a safe and effective technique for instrumenting the upper thoracic spine.

Objective To investigate the clinical and renal pathological features in the new rat model induced by anti-human podocyte-protein antibody. Methods The rat model was induced by once intravenous injection of rabbit anti-human podocyte-protein antiserum which was prepared at first. Male Sprague-Dawley rats (n=36) were randomly divided into six groups (6rats in each group): control group (CG), the time points of day 7 group (D7), day 14 group (D14),day 21 group (D21) and day 28 (D28) group after antiserum injection, and day 28 group after the normal rabbit serum injection (NRG). The level of 24 hour proteinuria, the clearance of creatinine,albumin, blood urea nitrogen, triglyceride, cholesterol and serum creatinine were measured. The renal morphology was detected under the light microscope, immunofluorescence microscope, and electron microscope. Results 24-hour proteinuria (mg) was gradually increased, and the level of proteinuria in D28 (48.56±13.80) was significantly higher than that in CG (5.34±2.77, P＜0.01)and NRG (11.32±4.90, P＜0.01). The clearance of creatinine (ml/min) and serum creatinine (μmol/L) in D28 (0.90±0.47, 33.48±9.94) were significantly different from CG (1.68±0.54, P＜0.05;26.03±2.67, P＜0.05), but showed no difference with NRG (1.34±0.87, P＞0.05; 27.40±4.73, P＞0.05). The level of albumin (g/L) was lower in D7, D14, D21, D28 (28.20±0.87, 27.80±1.97,27.42±1.66, 27.77±1.95) than CG (29.98±0.76, P＜0.05). But there was no difference in the level of albumin among the groups after antiserum injection and NRG (28.68±1.18, P＞0.05). The level of blood urea nitrogen, triglyceride, cholesterol showed no difference among the groups (P＞0.05). The renal morphology showed no obvious changes between CG and NRG. Among the groups after antiserum injection, the renal pathological changes under the light microscope were some spikes formation in D28. Immunostained for rabbit IgG in rat glomeruli progressively decreased over the 28 days, while rat IgG progressively increased. The renal section deposition for rat complement 3 reached a maximum at day 21 then decreased afterward. Under the electron microscope, there were immune complexes and foot process fusion at day 14. Conclusions The new rat model induced by anti-human podocyte-protein antibody showing typically clinical and pathological changes of the membranous nephropathy is successfully established.