Objective To provide a simple, convenient, and safe anesthesia method for the establishment of a M1 segment of middle cerebral artery occlusion model in rhesus monkey or other large laboratory animals.Method Twenty male rhesus monkeys weighing 7-11 kg (ages 7-9 years) from Academy of Military Medical Sciences were used in this study.Sumianxin injection combined with 0.1 mg/kg ketamine was given before endotracheal intubation (ID:4.5-5.5#).The animals were then transported to an interventional operation room, where the intravenous access was established and a urinary catheter was inserted into the urinary bladder.Mechanical ventilation was used during the surgery, propofol was continuously injected in a speed of 2-4 mg/kg/h, and Sumianxin-ketamine could be given if necessary to maintain adequate anesthesia depth.The dose was adjusted according to vital signs of the rhesus such as body movements, physiological parameters, and demand of surgery.Brain MRI examination was performed before and after thrombolysis.Anesthetic injection was suspended and the animals were allowed to have a spontaneous breathing every time before the MRI text.Heart rates, temperature, non-invasive blood pressure, and SpO2 were monitored during the whole surgery.Blood samples were taken from the radial artery for blood gas analysis after anesthesia induction and during operation.Results All the 20 animals underwent the operation successfully, no animal had restlessness, respiratory depression, arrhythmia and other serious complications.At the end of the study, animals awake soon.Fifteen of them survived longer than 24 hours, only 5 died from serious cerebral hemorrhage and larger cerebral infarction.Conclusions General endotracheal anesthesia is safe for rhesus monkeys during such interventional operation and MRI examination.

BACKGROUND:n mandibular posterior dental implantation,injury to the inferior alveolar nerve sometimes occurs because of mandibular canal going across mandibular body.This restricts the use of dental implantation at this site.Therefore,it is essential to understand the anatomic structure of inferior alveolar nerve canal in mandibular posterior dental implantation.OBJECTIVE:To observe the intramandibular course of and anatomic structure of inferior alveolar nerve canal.METHODS:Fifteen adult complete mandible specimens with teeth and 4 fresh mandible arterial infusion specimens were researched.All the specimens had complete dentition and there were no obvious absorption in alveolar bone.The course of inferior alveolar nerve canal and its dimension including transverse and longitudinal diameters of mandibular canal and the distance between mandibular canal and mandible each side (superior,inferior,buccal and lingual side) were measured in 15 adult mandibles with teeth.The relationship between blood vessels and nerve of the canal was observed in 4 fresh arterial infusion specimens.RESULTS AND CONCLUSION:The distance between the medial border of the mandibular canal and the lingual wall was shorter than that of the lateral wall of the mandibular canal to the buccal wall (P < 0.01);The length from the upper wall of mandibiular canal to the top of the alveolar ridge was longer than that of the inferior border of the mandibular canal to the inferior border of the mandible (P < 0.01).The longitudinal diameter was smaller than the transverse diameter (P < 0.05),namely,the cross section of the mandibular canal was an ellipse with a longer longitudinal diameter.There was no significant difference between the transverse and longitudinal diameters of the canal in the anterior and posterior teeth region of the mandible.The inferior alveolar nerve and its associated blood vessels were located within a nervous vascular bunch in the mandibular canals.In every fresh specimen the blood vessels lay above the nerve.There were small branches of blood vessels surrounding thenerve.The mandibular canal ran towards the lingual side and was close to the inferior margin of the mandible.

Objective To evaluate the relationship between inflammatory responses induced by perioperative infection and surgical stress and postoperative cognitive dysfunction in mice. Methods One hundred forty?four healthy male C57BL∕6 mice, aged 8-12 weeks, weighing 20-25 g, were divided into 4 groups ( n=36 each) using a random number table: control group ( group C) , surgery group ( group S) , infection group ( group I) , and infection+surgery group ( group I+S) . In group S, the open reduction and internal fixation was performed after tibial fracture was induced. Lipopolysaccharide ( LPS) 100 μg∕kg was injected intraperitoneally at the same time every day for 5 consecutive days starting from 1 day before surgery in group I. In group I+S, LPS 100 μg∕kg was injected intraperitoneally at the same time every day for 5 consecutive days starting from 1 day before surgery, and the open reduction and internal fixation was per?formed after tibial fracture was induced at 2 h after LPS injection on the day of surgery. Contextual fear con?ditioning test was performed on 1 and 3 days after surgery, and cognitive function was assessed. The rate of freezing time was calculated. The peripheral venous blood samples were collected for determination of plas?ma interleukin?6 ( IL?6) and IL?1β concentrations by enzyme?linked immunosorbent assay. The animals were then sacrificed, and the hippocampi were isolated for determination of IL?6, IL?1β and prostaglandin E2 ( PGE2 ) contents in hippocampal tissues by enzyme?linked immunosorbent assay. Results Compared with group C, the rate of freezing time was significantly decreased on 1 and 3 days after surgery, and the contents of IL?6, IL?1βand PGE2 in hippocampal tissues were significantly increased on 1 and 3 days after surgery in S and I+S groups, the concentrations of plasma IL?6 and IL?1βwere significantly increased on 1 day after surgery, and the concentration of plasma IL?1βwas significantly increased on 3 days after surgery in group S, the concentrations of plasma IL?6 and IL?1β were significantly increased on 1 and 3 days after surgery in I and I+S groups ( P<0?01) , and no significant change was found in the rate of freezing time on 1 and 3 days after surgery in group I ( P>0?05) . Compared with group S or group I, the rate of freezing time was significantly decreased on 1 and 3 days after surgery, and the concentrations of IL?6 and IL?1βin plasma and contents of IL?6, IL?1β and PGE2 in hippocampal tissues were significantly increased on 1 and 3 days after surgery in group I+S ( P<0?01) . Conclusion Inflammatory responses induced by periopera?tive infection and surgical stress can aggravate postoperative cognitive dysfunction in mice.

Objective To evaluate the efficacy of oxycodone or hydromorphone combined with propofol for colonoscopy.Methods A total of 150 patients of both sexes,aged 18-64 yr,weighing 45-85 kg,of ASA physical status Ⅰ or Ⅱ,undergoing colonoscopy,were randomly divided into 3 groups (n=50) using a random number table:fentanyl combined with propofol group (group F),oxycodone combined with propofol group (group O) and hydromorphone combined with propofol (group H).In F,O and H groups,fentanyl 1 μg/kg,oxycodone 0.1 mg/kg and hydromorphone 0.02 mg/kg were injected over 60 s,respectively,and then propofol 1.5 mg/kg was injected intravenously.After eyelash reflex disappeared,a colonoscope was placed.When body movement occurred during examination,half of the initial dose of propofol was added.The time for induction of anesthesia,operation time,emergence time,recovery time,occurrence of adverse cardiovascular events,nausea and vomiting and respiratory depression,and amount of propofol consumed were recorded.Results There was no significant difference between the three groups in the time for induction of anesthesia,operation time,emergence time,recovery time,adverse cardiovascular events,respiratory depression,and amount of propofol consumed.Compared with group F,the incidence of nausea and vomiting and respiratory depression was significantly decreased,and the degree was reduced in H and O groups.No significant difference was found between group O and group H in the incidence of nausea and vomiting and respiratory depression and the degree.Conclusion Oxycodone or hydromorphone combined with propofol can be safely and effectively used for colonoscopy and the efficacy is better than that of fentanyl combined with propofol.

AIM: To construct human Egr-1 promoter luciferase reporter system and study its activity induced by i-onizing radiation. METHODS: Egr-1 promoter was obtained by human genomic PCR and cloned into pGL3-basic vector. After transfection of recombinant plasmid into human tumor cells, the Egr-1 promoter activity induced by ionizing radiation was detected by luciferase reporter assay. RESULTS: The luciferasy reporter system of Egr-1 promoter was successfully constructed. The activity of Egr-1 promoter was substantially increased after different doses of IR and reached to the peak at the time point of 48h after IR. CONCLUSION: The Egr-1 promoter was constructed in this study showed IR inducible activity in tumor cells, laying foundation for the research of radiation, mediated gene therapy.

OBJECTIVETo reconstruct mandibular defect using tissue engineering bone with bone marrow stem cells (BMSCs) cell sheets and investigate the effect of cell sheets on osteogenesis.

METHODSBMSCs were isolated with the method of density gradient centrifugation from canine and cultured. BMSCs were induced to differentiate to osteoblasts. BMSCs induced were fabricated to BMSCs cell sheets. The poly (lactic-co-glycolic acid) (PLGA) wrapped with cell sheets were implanted into the mandibular defect in the left side (experimental side). PLGA wrapped without cell sheets were implanted into the right side (control side) of mandibles. 16 dogs were evenly divided into 4 groups, and one group of them was executed in 4, 8, 12, 16 weeks for gross investigation and histological observation.

RESULTSThe osteogenesis of experimental side was better than that of control side. 16 weeks after implantation, most areas of the mandibular defect were replaced by fresh bone tissue. Compact bone similar to normal bone tissue formed in the lingual defect of mandible and had bony union with the bone stump. The optical density of the fresh bone in the experimental side was higher than that of the control side, there was a significant difference between the two methods (P<0.05). Plenty of lamellar bones formed in experimental side and Haversian system, as well as red marrow, were observed.

CONCLUSIONTissue engineering bone with the structure of lamellar bones can be formed by the technology of BMSCs cell sheets.

Animals ; Bone Marrow Cells ; Bone and Bones ; Dogs ; Lactic Acid ; Mandible ; Osteoblasts ; Osteogenesis ; Polyesters ; Polyglycolic Acid ; Polymers ; Tissue Engineering

OBJECTIVETo construct functional tissue-engineered bone with cell sheet technology and method of traditional bone tissue engineering.

METHODSCanine bone marrow mesenchymal stem cells (BMSCs) were isolated with the method of density gradient centrifugation and cultured. BMSCs were induced to differentiate into osteoblasts and cultured in temperature-responsive culture dishes at 37 degrees C, 5% CO2 and saturated humidity. BMSCs cell sheet was prepared when temperature was changed to 20 degrees C. Demineralized bone matrix (DBM) and platelet-rich plasma (PRP) were prepared, and complex of DBM/PRP/BMSCs cell sheet/BMSCs was construsted and implanted under the left latissimus dorsi muscle. Complex of DBM/PRP/BMSCs was implanted under the right latissimus dorsi muscle.

RESULTSWhen temperature dropped at 20 degrees C, BMSCs detached automatically from the temperature-responsive culture dishes and formed an intact cell sheet. The osteogenesis of the DBM/PRP/BMSCs cell sheet/BMSCs group was better than that of the DBM/PRP/ BMSCs group.

CONCLUSIONCell sheet technology combined with traditional bone tissue provides a new way for construction of ideal functional tissue-engineered bone.

Animal Experimentation ; Animals ; Bone Marrow Cells ; Bone and Bones ; Cells, Cultured ; Mesenchymal Stromal Cells ; Osteoblasts ; Osteogenesis ; Platelet-Rich Plasma ; Stromal Cells ; Tissue Engineering

Objective To summarize the postoperative complications of reconstruction of mandible defect with titanium reconstruction plate.Methods A total of 111 cases of the mandibular defect caused by various reasons and repaired by titanium reconstruction plate in the Department Oral and Maxillofacial Surgery of the affiliated Hospital of Qingdao University from 2003 to 2012 were collected and followed up.The complications were analyzed.Results Thirty-seven percent of 111 cases showed long term complications.The titanium fracture was the main complication(16％[18/111]),followed by stress-shielding (9％[10/111]),infection(8％[9/111]),and titanium plate exposure(4％[4/111]).Titanium plate fracture occurred within 8 months and 3 years after surgery.The simple titanium plate reconstruction had the highest rate of plate fracture(30％[15/50]).Stress-shielding in non-vascularized bone graft was more significant than that in vascularized bone graft(P＜0.05).When replaced by mini-titanium plate,the stress-shielding effect disappeared gradually.When the retention of mandibular margin height was less than 1 cm with the use of reconstruction plate,the postoperative complications were prone to occur.Conclusions Bone graft is the best way to reconstruct mandibular defect,and simple reconstruction plate repair is applied only as a transitional means for high degree of malignancy,obvious recurrence tendency tumor or special reasons such as age etc,which are not suitable for bone graft.The reconstruction plate fixation is not recommended for bone graft,especially non-vascularized bone graft.The retention of mandibular margin with reconstruction plate fixation is open to discussion.

Objective:To observe the neuroprotective effect of wind medicine (Puerariae Lobatue Radix and Acori Tatarinowii Rhizoma) in combination with Panax ginseng total saponin (PNS) on cerebral ischemia-reperfusion rats; To elucidate the mechanism of "wind medicine increasing effect".Methods:Totally 140 male SD rats were divided into sham-operation group, model group, PNS group, Puerariae Lobatue Radix group, Acori Tatarinowii Rhizoma group, Puerariae Lobatue Radix + PNS group, Acori Tatarinowii Rhizoma + PNS group according to the random number table method, with 20 rats in each group. Except for the sham-operation group, the cerebral ischemia/reperfusion rat model was established using the modified Longa line bolus method in the remaining groups. After 7 d of administration of the appropriate pharmacologic intervention in each group, neurological dysfunction was evaluated by Zea-longa score after final administration, cerebral infarct volume was determined by TTC staining; blood brain barrier (BBB) permeability of brain tissue on the ischemic side was detected by Evans blue content; BBB ultrastructure of each group of rats was observed by transmission electron microscopy; Claudin 5 protein expression level was detected by immunohistochemistry; Zonula occludens-1 (ZO-1), major facilitator supeffamily domain-containing protein 2a (Mfsd2a), Occludin, P-glycoprotein (P-gp), Monocarboxylate Transporters-1 (MCT1) and breast cancer resistance protein (BCRP) protein expression levels were detected by Western-blot.Results:Compared with the model group, the rat neurological function scores were reduced in each administration group ( P<0.05), infarct volume was reduced ( P<0.05), EB content of brain tissue decreased ( P<0.05), protein expressions of Claudin 5, ZO-1, Mfsd2a and Occludin in brain tissue were elevated ( P<0.05), the protein expressions of P-gp, BCRP and MCT1 were reduced ( P<0.05), and the protein expressions of Claudin 5, Mfsd2a, and Occludin was higher in the Puerariae Lobatue Radix + PNS group and Acori Tatarinowii Rhizoma + PNS group than that of each group of medication alone ( P<0.05), and the protein expression of MCT1 was lower than that of each group of medication alone ( P<0.05); the protein expression level of ZO-1 in the Puerariae Lobatue Radix + PNS group was higher than that of the group of medication alone ( P<0.05); P-gp protein expression was lower in Acori Tatarinowii Rhizoma + PNS group than in the PNS group and Acori Tatarinowii Rhizoma group ( P<0.05). Conclusion:Wind medicine (Puerariae Lobatue Radix and Acori Tatarinowii Rhizoma) may potentiate the neuroprotective effect of PNS on cerebral ischemia-reperfusion rats, and the mechanism may be related to the protection of BBB structural integrity and maintenance of central barrier properties, while regulating substance transport proteins and increasing the intracerebral content of the drug.

Using mutation PCR, we cloned the target gene containing 421-480nt (141-160aa) and 598-639nt (200-213aa) of VP1 gene of foot and mouth disease virus (FMDV) into the deleted region (508-532aa) of Nsp2 gene of a highly pathogenic porcine reproductive and respiratory syndrome virus derived vaccine strain (HuN4-F112) that was used as vector. The recombinant cDNA was in vitro transcribed followed by transfection of BHK-21 cells for 36 h. Then, the supernatant of the cell culture was continuously seeded to monolayer of MARC-145 cells for recovery of the recombinant virus. CPE was obviously visible after a couple of passages in the seeded MARC-145, and the rescued virus (designated as rPRRSV-F112-O/VP1ep) was identified by Mlu I digestion, sequencing and immunofluorescence assay. Meanwhile, expression of inserted FMDV epitopes was also detected by indirect immunofluorescence assay with polyclonal antibodies against VP1 protein of FMDV. The analysis of biological characteristics shows that the titer of the rescued recombinant PRRSV (TCID50 = -log10(-6.75)/0.1 mL) was similar to its direct parental virus rHuN4-F112-delta508-532, but higher than rHuN4-F112.
Animals ; Antigens, Viral ; immunology ; Base Sequence ; Capsid Proteins ; immunology ; Cell Line ; Cysteine Endopeptidases ; genetics ; Epitopes ; genetics ; Foot-and-Mouth Disease ; immunology ; prevention & control ; Foot-and-Mouth Disease Virus ; genetics ; immunology ; Molecular Sequence Data ; Mutation ; Porcine respiratory and reproductive syndrome virus ; genetics ; immunology ; Recombination, Genetic ; Swine ; Transfection ; Vaccines, Attenuated ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; immunology ; Viral Vaccines ; genetics ; immunology