Objective To investigate the effect of pregnancy on the potency of bupivacaine for spinal anesthesia in rats. Methods Female non-pregnant SD rats weighing 180-220 g and 17 day pregnant SD rats weighing 350-400 g were used in this study. The rats ( 18 non-pregnant, 18 pregnant) in which PE-10 catheter were successfully placed without complications were selected. The 18 non-pregnant rats were randomly divided into 3 groups (n =6 each): control group (group C), 2% bupivacaine group (group B2 ) and 4% bupivacaine group (group B4). The 18 pregnant rats were also randomly divided into 3 groups (n = 6 each): control group (group PC),2% bupivacaine group (group PB2 ) and 4% bupivacaine group (group PB4 ). Group C and PC received intrathecal (IT) normal saline 30 μl, and the other 4 groups received 2% or 4% bupivacaine 30 μl intrathecally. Analgesia was determined using the taifllick latency (TFL) before IT administration (baseline), and at 10 min, 20 min,30 min, 1 h, 2 h, 4 h, 1 d, 2 d, 3 d and 4 d after IT administtation. The percentage of the maximal possible effect (MPE) was calculated. Hind-limb motor function (MF) was also assessed. Results Compared with the baseline value, MPE at 10 min-2 h after administration and MF scores at 10 min-1 h after administration were significantly increased in group B2, MPE at 10 min-4 h after administration and MF scores at 10 min-1 h after administration were significantly increased in group B4;MPE at 10 min-1 d after administration and MF scores at 10 min2 h were significantly increased in group PB2 and MPE at 10 min-1 d after administration and MF scores at 10 min4 h were significantly increased in group PB4 ( P ＜ 0.05 ). Conclusion Pregnancy can enhance the potency of bupivacaine for spinal anesthesia in rats.

BACKGROUND:Bladder acellular matrix graft (BAMG) is frequently used for domains of tissue engineering scaffold due to its great biocompatibility and cell adhesion. OBJECTIVE:To verify the biological characteristics of BAMG after frozen and lyophilized processing. DESIGN,TIME AND SETTING:A biocompatibility experiment was performed at Shanghai Tissue Engineering Research and Development Center and Experimental Animal Department of the Sixth People's Hospital of Shanghai between May and November 2008. MATERIALS:Two New Zealand rabbits were used in this study for BAMG preparation. METHODS:After midsection of rabbit bladder,mucous membrane of urinary bladder was isolated and dipped in three-distilled water for 24 hours. Thereafter,the samples were incubated with acellular solution containing 0.1% Triton X-100 and 0.15% aqueous ammonia for 14 days. The culture medium was changed regularly. The samples in the control group were stored in 75% ethanol,while samples in the experimental group were frozen for 24 hours at -80 ℃,vacuum-dried for 24 hours,and stored in 75% ethanol. MAIN OUTCOME MEASURES:Biological characteristics of BAMG were detected using hematoxylin and eosin staining,Masson staining,and scanning electron microscopy; biological characteristics were compared between the two groups using cell adhesion test,MTT assay,and subcutaneously embedding test. RESULTS:Hematoxylin and eosin staining and Masson staining revealed that no residual cells were detected in the BAMG,and collagen was intact. Scanning electron microscopy demonstrated that cells exhibited a slit-shaped structure mainly containing collagen which was beneficial for cell adhesion. Mechanical test revealed that the BAMG after frozen and lyophilized processing not only reserved the mechanical properties of the raw BAMG,but also had a great elongation. MTT assay confirmed that cytotoxicity was grade 0,and BAMG had a good compatibility to smooth muscle cells. After subcutaneously embedding for one month,BAMGs had good adhesions to subcutaneous tissues,and muscular adhesion and vascular proliferation were observed. CONCLUSION:BAMG after frozen and lyophilized processing reserves original biocompatibility and has great elongation; therefore,it will become a useful and ideal biomaterial for tissue engineering scaffold.

Objective To explore the targeted nursing care for patients with severe traumatic brain injury complicated with intracranial infection. Methods 35 patients of severe traumatic brain injury complicated with intracranial infection from January, 2009 to December, 2011 were reviewed. Results There were 10 cases cured, 12 cases improved very well, 4 cases improved, 5 cases invalid and 4 cases died. The total effective rate was 62.86%and the fatality rate was 11.43%. Conclusion Intracranial infection can be controlled effectively by the venous and intrathecal injection of antibiotics and targeted care in the patients with severe traumatic brain injury.

BACKGROUND: The application of adipose derived stromal cells to tissue engineering has been more and more popular around the world. Compatibility of scaffold material is the key point for its further research.OBJECTIVE: To determine the growth rules of rabbit adipose-derived stromal cells with polylactic-co-glycolic acid (PLGA).DESIGN, TIME AND SETTING: An in vitro study was performed at the Institute of Ophthalmology, Otolaryngology Hospital,Fudan University and Shanghai Tissue Engineering Center from September 2007 to March 2009.MATERIALS: Six female New Zealand rabbits aged six months were used for extraction of adipose-derived stromal cells. PLGA was provided by Sigma, USA.METHODS: Adipose tissue was harvested from the nape fat pad of the rabbits following anesthesia. Primary cultured cells were established using type I collagenase and cell cultures were maintained with DMEM containing 10% volume fraction of fetal bovine serum. Cells were passaged when 80% was confluent. The fourth passages of cells were utilized for the study. PLGA consisted of polylactic acid and polyglycolic acid as the ratio of 7:3, and the relative molecular mass was 104900.MAIN OUTCOME MEASURES: Initially, the adherent rate of cells to scaffold was detected. After one week co-culture, the scaffold bearing adipose-derived stromal cells labeled with Dio agent were investigated by fluorescence inverse microscope,scanning electron microscopy and laser scanning microscope.RESULTS: The best adherent rate of adipose-derived stromal cells with PLGA reached 99%. After one-week-incubation in vitro,cells of fiber-shaped or ovule-shaped proliferated well and exhibited stratified growth on the surface of PLGA scaffold. In addition,it also secreted visible extracellular matrices, which could be examined by scanning electron microscopic examination.Meanwhile, the adipose-derived stromal cells grew well and distributed equably inside the PLGA in terms of the investigation with laser scanning microscope.CONCLUSION: The compatibility of adipose-derived stromal cells to PLGA in vitro was well.

Objective To evaluate the efficacy of decortication by video-assisted thoracic surgery (VATS) in pa﹣tients with tuberculous empyema, and discuss its indications. Methods 60 patients with tuberculous empyema who underwent decortication by VATS for surgical management from December 2010 to December 2015 were included. Under a thoracoscope, we cleaned up the pus, separated adhesions, scraped granulation tissues and caseous necrosis on the inner wall of the abscess cavity, and stripped the thickened fiberboard of the parietal and visceral pleurae. Af﹣ter the procedure, sufficient drainage and antituberculosis therapy were carried out. Results All the patients in this group were operated successfully. All the patients were cured without perioperative death and complications. No re﹣currence of empyema was observed at the follow-up examination from 2 months to 5 years, and suffered pulmonary reexpansions were better. Conclusions The decortication by VATS for tuberculous empyema is safe, effective, mini﹣mally invasive. The imaging manifestations of pleural thickening in 1 cm, no obvious calcification, no serious lesions in the lungs are the indications for the operation.

Objective To explore the method of culture of rat adipose-derived stem cells(ADSCs) and differentiation induction into myoblasts. Methods Adipose tissues were obtained from SD rats,and were isolated by enzyme digestion and cultured into ADSCs.The expression of surface antigen CD90,CD105 and CD34 was detected by immunofluorescence and flow cytometry.ADSCs of the second passage with logarithmic growth were obtained,and culture media containing 5-azacytidine(5-aza) and basic culture media were employed for cells in induction group and control group,respectively.The induction lasted for 7 d,14 d,21 d,28 d and 35 d,respectively.Cell growth and cell morphology were observed by inverted phase contrast microscope,and immunofluorescence and flow cytometry were utilized to detect the expression of myoblast specific antigens desmin and myosin. Results ADSCs were successfully isolated and cultured,and were identified to be stem cells.On the 28th day of induction,cells in induction group displayed "swirl" morpholgy,and multinucleation was observed.It was revealed by immunofluorescence and flow cytometry that the highest expression rates of desmin and myosin were 52.57% and 50.04%,respectively on the 28th day of induction,while there was no expression before induction and in control group. Conclusion ADSCs can be isolated and cultured from rat adipose tissues,and can further differentiate into myoblasts after induction by culture media with 5-aza.The expression of myoblast specific antigen is the highest on the 28th day of induction.

BACKGROUND:Alveolar bone remodeling and sustained absorption due to tooth extraction seriously affect the implanting conditions and morphology of hard and soft tissue in implant zone. OBJECTIVE:To evaluate the effect of nano-hydroxyapatite/polycaprolactone electrospinning scaffolds to improve the osteogenic effect of bone defects around immediate implants. METHODS:Tissue-engineered bone was prepared by combining canine bone marrow mesenchymal stem cels with nano-hydroxyapatite/polycaprolactone electrospinning scaffold. Bilateral mandibular second premolars from six dogs were extracted mandibular second premolar, and an immediate implant was placed in the mesial fossa of the mandibular second premolar. Three-wal bone defects was made buccaly using titanium nails, then tissue-engineered bone and Bio-Oss bone powders were implanted bilateraly covered by colagen membranes (Bio-Gide). Imageology examination was performed to measure bone gray levels immediately, 4, 8, 12 weeks after surgery. After 12 weeks, the mandible was removed completely, toluidine blue staining was used for observation of microstructure, new bone formation, bone morphology and implant osseointegration. RESULTS AND CONCLUSION: Between the two groups, there was no difference in bone mineral density at each time point after surgery, indicating that the effects of the two materials to promote bone regeneration process are basicaly the same. After implantation, the dense lamelar bone formed in the bone defect region of tissue-engineered bone group, mature bone cels, Haversian canal, and implant osseointegration were visible. While, in the Bio-Oss group, the lamelar bone was dense, a smal amount of Bio-Oss particles distributed within new bone tissues, fewer bone cels were found, a part of Haversian canal was shown to have blood capilaries, and new bone was in close conjunction with the implant. These findings indicate that the nano-hydroxyapatite/polycaprolactone electrospinning scaffold combined with bone marrow mesenchymal stem cels and Bio-Gide colagen membrane can promote the regeneration of alveolar bone around the implant.

BACKGROUND:Fibrin glue introduced into calcium phosphate cement has not been confirmed whether this way could overcome the compressive limits and the low degradation of calcium phosphate cement and to modify the biological properties of calcium phosphate cement. OBJECTIVE:To explore the mechanical and biological properties of calcium phosphate cement/fibrin glue at different powder/liquid ratio for bone regeneration in vitro. METHODS:Calcium phosphate cement and fibrin glue were mixed at ratios of 1:1, 3:1, 5:1 (mL/g), and the pure calcium phosphate cement served as controls. Setting time, scanning electron microscope and the biomechanical test were used to analyze the composite scaffold structure, physical performance and the mechanical properties. Passage 3 osteoblasts were respectively inoculated on the material surface of the four groups, and pure cells served as blank controls. celladhesion, proliferation and alkaline phosphatase activity were observed. RESULTS AND CONCLUSION:The initial and final setting time of calcium phosphate cement/fibrin glue at 1:1 and 3:1 (mL/g) was higher than that in the control group (P<0.05), while the initial and final setting time of calcium phosphate cement/fibrin glue at 5:1 (mL/g) was lower than that of the control group (P<0.05). Scanning electron microscope showed smoother and denser surface of composite scaffolds compared with the pure calcium phosphate cement. The aperture of the composite scaffolds was decreased with the increasing concentration of fibrin glue. The compressive strength of composite scaffolds at 3:1 and 5:1 was higher than that of the control group (P<0.05), while the modulus of the composite scaffolds at 1:1, 3:1, 5:1 was higher than that of the control group (P<0.05). celladhesion, proliferation and alkaline phosphatase activity showed no difference among the three composite scaffold and control groups, but al higher than the blank control group (P<0.05). These findings indicate that fibrin glue introduced into calcium phosphate cement can overcome the low-strength limits of calcium phosphate cement, and maintain the good biological properties of calcium phosphate cement for bone regeneration.

Objective To investigate and assess the best seeding method for constructing three dimensional urethral tissue in vitro. Methods High speed agitation decellular method was used for preparing the porcine acellular corporous spongiosum matrix (ACSM). Before seeding, the matrix was sterilized via soaking compound iodine solution. Rabbit tongue epithelial cells and cavernosal smooth muscle cells were isolated and cultured. Three different groups of seeding method was used in this study. Group A (sandwich seeding group): The smooth muscle cells and epithelial cells were seeded onto the different side of ACSM by static method. Group B (injection seeding group): The smooth muscle cells were injected into the scaffold. Then, epithelial cells were seeded onto the urethral surface of ACSM by static method. Group C (agitation seeding group): The smooth muscle cells were seeded into the scaffold by agitation method. Then, epithelial cells were seeded onto the urethral surface of ACSM by static method. After being seeded, all matrixes were cultured in vitro for 14 d. HE and immunoassay staining were used to examine the results of seeding. Results Looser matrix was obtained after using high speed agitation decellular method. Compound iodine solution could not only sterilize efficiently but also reserve the original structure of biomaterial. An intact epithelial cellular layer onto the surface of scaffold could be observed in HE staining section after 14 d culturing in vitro.Few smooth muscle cells could be found in big space of biomaterial in group A. In group B, smooth muscle cells were restrained in some regions of the matrix. Smooth muscle cells were well distributed into the scaffold in group C. Conclusions After using high speed agitation decellular method, an ideal matrix with three dimensional structure can be obtained. Combined with agitated seeding method, three dimensional urethral tissue can be constructed.

Objective To explore the cultural method of oral keratinocytes and make preparations for further investigation in using oral keratinocytes as a new choice of seed cells for the reconstruction of tissue-engineered urethra. Methods Oral keratinocytes of rabbits were isolated in vitro and seeded onto a culture dish with a feeder layer of 3T3 mouse fibroblasts inhibited by mitomycin(i3T3) or a culture dish without i3T3 respectively. Cell morphology was observed and cell growth was detected at intervals.Meanwhile,oral keratinocytes obtained from in vitro culture were performed immunofluorescence staining with broad-spectrum keratin antibody(AE1/AE3) and keratin 19 antibody(K19).The percentage of positive cells of passage 2 reactive to AE1/AE3 was assessed by flow cytometry. ResultsOral keratinocytes seeded onto a feeder layer of i3T3 exhibited finer morphous,better amplification capability,and could be passed for 7 or 8 generations.However,those cultured without i3T3 took on various morphous and could only be subcultured 2 generations before ageing.It was indicated by immunofluorescence staining that oral keratinocytes obtained from in vitro culture were positive for AE1/AE3 staining and 40% were positive for K19 staining.The result of flow cytometry revealed that the amout of positive keratinocytes reactive to AE1/AE3 was more than 95% of total cellular score. Conclusion Oral keratinocytes of rabbits can be cocultured with i3T3 in vitro and magnitude quantity can be attained,laying a favourable foundation for oral keratinocytes as a new choice of seed cells for urethral reconstruction with tissue engineering.