BACKGROUND: Soft tissue filling is a problem in clinic. It has been proved that ultrasonic wave-treated mature adipocytes cannot be used for cell transplantation.It is hopeful to solve the problem with adipose tissue engineering. OBJECTIVE: To observe the effect of ultrasonic wave on the in vitro culture and proliferation of preadipocytes, and validate the possibility of preadipocyte as seed cell in adipose tissue engineering following ultrasound-assisted liposuction. DESIGN: Controlled observation experiment. SETTING: Department of Plastic Surgery, Rennin Hospital, Wuhan University. MATERIALS: This experiment was carried out in the Medical College of Wuhan University from July 2003 to September 2004. One local 3-month-old hybridized pig was provided by the Experimental Animal Center of Medical College of Wuhan University. After the pig was anesthetized with ketamine, an area of 10 cm×20 cm was labeled on both sides of back respectively, and the tissue in the labeled area was harvested. The tissue on the right side served as experimental group and that on the left side served as control group. METHODS: The tissue of the expedmental group was pretreated for 8 minutes by ultrasonic wave with the energy of 3W/cm2. Under aseptic condition, the skin layer was open, and 100 g subcutaneous adipose tissue was resected from each side and then placed in prepared container containing cold D-hanks solution for later use. The ultrasonic wave-treated porcine preadipocytes of adipose tissue of experimental group were isolated and cultured in vitro, and the number and lipid content of preadipocytes were measured every other 2 days. Results were presented as the mean val ue of the number of cells of three wells. In the control group, pretreatment of ultrasonic wave was omitted. The growth curves of two groups were drawn. Intracellular adipose content was measured by oil red O staining. Absorbance (A) was measured with spectrophotometer (HITACHI G2000), which was regulated at the wavelength of 510 nm. MAIN OUTCOME MEASURES: ① Morphological observation of culture of porcine preadipocytes. ②The growth curves of experimental and control groups. ③3 Change in the lipid content of experimental and control groups.RESULTS: ① Cells in the control group adhered to the wall within 6 to 24 hours, and those in the experimental group basically adhered to the wall after 1 to 2 days, and there were many non-adhesive cells in the exoerimental group. Accumulation of adipose granules in the cells was found in the control group after 4 days and that was found in the experimental group after 6 days. On day 12, the cells in the control group basically differentiated into the cells with single lipid drop or multiple lipid drops, and a small amount of natant mature adipocytes were found, while few cells with lipid drop were found in the experimental group. ②Under the condition of the same amount of cells, the cell doubling time of experimental group was about 72 hours and that of control group was 36 hours. After 9-day culture, the number of cells in the experimental group and control group was 13×104 cells/well and 18×104 cells/well, respectively. The rate of cell proliferation of experimental group was very slow. ③Lipid of the experimental group appeared on the 6th day after culture, and that of the control group appeared on the 4th day after culture. The peak value of A was 0.32 and 0.68 in e penmental group and control group respectively.CONCLUSION: Ultrasonic wave has obviously inhibitory effect on the proliferation and differentiation of preadipocytes.The preadipocytes treated by ultrasonic wave are not suitable as seed cells.

Objective To observe the effect of subarachnoid block with 0.5% bupivacaine at different temperatures during cesarean section.Methods 100 cases of elective cesarean section were randomly divided into room temperature group and heating group,50 cases in each group.Room temperature group: bupivacaine hydrochloride injection and glucose injection equilibrated group in a constant temperature thermostatic bath of 24 degrees thermostatic bath heating for above 30 min.Heating group: bupivacaine hydrochloride injection and glucose injection heated in the constant temperature thermostatic bath of 37 degrees thermostatic bath heatingfor above 30 min.Anesthesia was injected into the subarachnoid space at different temperatures to observe the anesthetic effect.Results The anesthesia increased rapidly, and the analgesia and muscle relaxation effects were better in the heating group than room temperature group, but the heating group had hypotension rate was higher than the room temperature group (36.0% vs.16.0%).There was no obvious difference between the incidence of adverse reactions such as nausea and vomiting in both groups.Conclusion Different temperatures of bupivacaine can be used safely for section anesthesia.The anesthesia effect of the heateding bupivacaine is faster, the anesthesia level is higher, the anesthesic and muscle relaxant effect is better.Bupivacaine at room temperature has relatively small effect on hemodynamics.

BACKGROUND:Operation is an important measure to improve the function of spinal cord and to stop the pathological progress of multilevel cervical spondylotic myelopathy. There are controversies how to select the optimum operation mode, to reduce postoperative complications and to elevate clinical curative effects.
OBJECTIVE:To systematical y review patients’ profiles of multilevel cervical spondylotic myelopathy, and to evaluate the effects of simple anterior approach, simple posterior approach and one stage posterior anterior combined approach on cervical spinal curvature index and functional recovery in patients.
METHOD148 sample profiles of patients, who received multilevel cervical spondylotic myelopathy operation in The Affiliated Hospital of Qingdao University and Qingdao Municipal Hospital from February 2000 to February 2008, and met the inclusion and exclusion criteria, were selected. They were divided into simple anterior approach group, simple posterior approach group and one stage posterior anterior combined approach group. The differences in the functional recovery were assessed after treatment using different therapeutic methods.
RESULTS AND CONCLUSION:Cervical spinal curvature index was highest in the simple posterior approach group before treatment (P<0.01). Cervical spinal curvature index was highest in the one stage posterior anterior combined approach group after treatment (P<0.01). Changes in cervical spinal curvature index were most obvious in the simple anterior approach group before and after treatment (P<0.01). No significant difference in Japanese Orthopaedic Association Scores was detected among three groups after treatment (P>0.05).
Significant differences in improvement rate of Japanese Orthopaedic Association Scores were detectable after treatment between the one stage posterior anterior combined approach group and simple anterior approach and simple posterior approach groups (P<0.001). Significant differences in cervical dysfunction index and SF-36 scores were detectable among the three groups before and after treatment (P<0.05). Results indicated that compared with the simple anterior approach and simple posterior approach, decompression through one stage posterior anterior combined approach is a reliable and effective operative procedure for treatment of multilevel cervical spondylotic myelopathy.

BACKGROUND:It has been confirmed that carboxymethylated chitosan has an promoting effect on Schwann cel proliferation and secretion, but its impact on the cyclic adenosine monophosphate-mediated protein kinase A signaling pathway in schwann cel stil needs further study. OBJECTIVE:To investigate the effect of carboxymethylated chitosan on cyclic adenosine monophosphate/ protein kinase A signaling pathway in rat schwann cels. METHODS:The Schwann cels of the second generation neonatal rats were obtained and seeded in 6-wel plate at a concentration of 1×109/L. These Schwann cels were cultured and divided into four groups. The Schwann cels in the control group were cultured by adding PBS. The Schwann cels in the experimental groups were cultured by adding 50, 100 and 200 mg/L of carboxymethyl chitosan solution, respectively. After 24 hours, the concentration of cyclic adenosine monophosphate, protein kinase A activity and cyclic adenosine monophosphate response element binding protein mRNA expression were detected. RESULTS AND CONCLUSION:Compared with the control group, carboxymethyl chitosan increased cyclic adenosine monophosphate concentrations, the activity of protein kinase A and cyclic adenosine monophosphate response element binding protein mRNA expression within the Schwann cels in a dose-dependent manner (P < 0.05). These results demonstrate that carboxymethyl chitosan can increase the concentration of cyclic adenosine monophosphate within the Schwann cels and promote protein kinase A activity, thereby activating cyclic adenosine monophosphate/protein kinase A signaling pathway.

Objective To compare the effects of a single injection of palonosetron and tropise-tron to prevent postoperative nausea and vomiting (PONV ) in gynecological surgeries. Methods Sixty patients undergoing elective major gynecological surgeries with general anesthesia (Apfel score ≥ 3 ) were included and randomized to group P (palonosetron ) and group T (tropisetron),30 patients in each group.All patients received general anesthesia with tracheal intuba-tion,and palonosetron 0.25 mg or tropisetron 5 mg were injected before anesthesia respectively in two groups.Intravenous hydromorphine was delivered for postoperative analgesia in all patients. PONV were evaluated and followed up for 72 hours.The degree of PONV was recorded and the com-plete response rate (CR)and complete control rate (CC)were calculated.Results The degree of PONV in 0-24 h and 24-48 h was milder in group P than in group T (P <0.05),while in 48-72 h the degree of PONV was similar between the two groups.In group P,5 patients had vomiting postopera-tively with the failure period of treatment of (1 9.6±9.4)h,and no patients needed rescue treatments. While in group T,1 9 patients suffered from vomiting with the failure period of treatment of (20.6± 4.5)h,and rescue treatments were delivered 3 times in total.The CR and CC of preventing PONV in 0-24 h,24-48 h and 0-72 h were significantly higher in group P than in group T (P <0.05).The CR and CC of the two groups were comparable in 48-72 h postoperatively.Conclusion A preoperative single dose of palonosetron 0.25 mg is better than tropisetron 5 mg in preventing PONV within 48hours after gynecological surgery.

ObjectiveTo observe the effect of penehyclidine hydrochloride (PHC) on serum interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α ) concentrations following tourniquet deflation in patients undergoing lower limb surgery.Methods Thirty adult patients,scheduled for unilateral lower limb surgery,ASA classification [ - Ⅱ grades,were divided into control group and research group by random number table,each group of 15 cases.Before anesthesia 30 min,PHC in intravenous infusion of 0.01-0.02 mg/kg in research group,the corresponding volume in intravenous infusion of 0.9% sodium chloride in control group.Peripheral venous blood samples were collected immediately before tourniquet inflation (T0,baseline),immediately before tourniquet deflation(T1),30 min (T2) and 60 min (T3) after tourniquet deflation.Serum IL-6 and TNF- α concentrations were measured by enzyme linked immunosorbentassay.ResaltsSerum TNF- α change at the same time point after tourniquet deflation was not statistically significant between two groups (P ＞ 0.05).Serum IL-6 concentration was decreased at each time point after tourniquet deflation compared with T0 in research group,while increased in control group.Serum IL-6concentration difference of T3 and T0 had statistically significant between research group and control group [ (-8.8 ± 5.6) ng/L vs.( 10.2 ± 6.7) ng/L,P＜ 0.05].ConclusionsPHC in advance can decrease serum IL-6 concentration after tourniquet deflation in patients undergoing lower limb surgery.

ObjectiveTo summarize and analyse the perioperative management especially theanesthesia of 20 kidney transplant recipients in non-transplant surgery. MethodThe anesthesia management of 20 kidney transplant recipients in non-transplant surgery was analyzed retrospectively. ResultsIn 20 cases, 1 case (5％) was performed under local anesthesia,4 cases (20％) were performed under intravertebral anesthesia and 15 cases (75％) were performed under general anesthesia. The operation time was 30-260 min, all cases were managed successfully. ConclusionIt is still a clinical challenge to deal with the surgical patients after kidney transplantation, and it needs fully understanding of the pathophysiological status of the patient and closely collaboration of transplant physicians, anesthesiologists and the surgeons.

Objective To study the effects of carboxymethylated chitosan (CMCS) to nitric oxide (NO)-induced apoptosis on rat chondrocytes,and explore p38MAPK signal transduction pathway in the process and its mechanism.Methods The rat articular cartilage cells were cultured in vitro,collagen type-2 (collagen-2) immunohistochemical staining was used to identify the cartilage cells.The model of chondrocyte apoptosis was built by different concentrations of sodium nitroprusside (SNP) induction.The cells were divided into the control group,the SNP treated group SNP+CMCS treated group,and the SNP+p38 MAPK inhibitor SB203580 treated group.The apoptotic rate of chondrocytes was calculated by FCM,apoptotic nuclei was identified by Hoechst33342 stain,the mitochondrial membrane potential changes was detected by Rhodamine123 (Rho123) stain,the expression of p38 and p-p38 were detected by Western blotting analysis.Results 1-3 mmol/L SNP could induce chondrocyte apoptosis,the apoptotic rate was increased with the SNP increasing,the most obvious apoptosis was occurred in 3 mmol/L SNP treated chondrocytes,which was 69.8％ (P＜0.05).SNP could increase the nuclear fragmentation of chondrocytes,the cells with nuclear fragmentation was significantly higher than that in the control group.SNP could reduce mitochondrial membrane potential in chondrocytes,which decreased significantly compared with the control group.SNP could increase the p-p38 expression in chondrocytes,which was 4.3 times compared to the control group.CMCS of different concentrations could reduce the apoptotic rate of SNP-induced chondrocytes,which was 51.0％,29.9％ and 15.2％,which was decreased significantly (P＜0.05) when compared with 3 mmol/L SNP induced group,CMCS decreased the cells number of SNP-induced nuclear fragmentation.CMCS increased the mitochondrial membrane potential in SNP-induced chondrocytes.CMCS reduced the expression levels of p-p38 in SNP-induced chondrocytes.Conclusion CMCS has protective effect on SNP-induced apoptosis of chondrocytes.This process is completed by inhibiting the activity of p38 MAPK signal pathway.

Objective To evaluate the effect of lithium chloride (LiCl) pretreatment on isoflurane-induced cognitive dysfunction and inflammatory response in hippocampus in aged rats.Methods Eighty 20-month-old male Sprague-Dawley rats,weighing 350-400 g,were randomly assigned into 4 groups (n =20 each):control group (group C),1.4％ isoflurane group (group I),100 mg/kg LiCI + 1.4％ isoflurane group (group L+ I),and 100 mg/kg LiC1 group (group L).Group I was exposed to 1.4％ isoflurane in 30％ O2-70％ N2 for 6 h,while group C was exposed to 30％ O2-70％ N2 only.LiCl 100 mg/kg was injected intraperitoneally once a day for 3 consecutive days and isoflurane anesthesia was performed on 4th day in group L + I.LiCl 100 mg/kg was injected intraperitoneally once a day for 3 consecutive days and then the rats inhaled 30％ O2-70％ N2 for 6 h on 4th day in group L.Blood samples were taken immediately after the end of anesthesia for blood gas analysis.Hippocampi were isolated 24 h after the end of anesthesia for determination of the expression of glycogen synthase kinase-3β (GSK-3β) and acetyl-NF-κB (Lys310) (by Western blot) and TNF-α,IL-1β and IL-6 mRNA (by RT-PCR).The levels of TNF-α,IL-1β and IL-6 were determined by ELISA and the contents of TNF-α,IL-1β and IL-6 were calculated.The cognitive function was assessed on 2nd day after the end of anesthesia.Results Compared with group C,the expression of GSK-3β and acetyl-NF-κB (Lys310) was significantly up-regulated,the expression of TNF-α,IL-1β and IL-6 mRNA and contents of TNF-α,IL-1β and IL-6 were increased,the escape latency was prolonged,and the time of staying at the original platform quadrant was shortened in group I (P ＜ 0.05).Compared with group I,the expression of GSK-3β and acetyl-NF-κB (Lys310) was significantly down-regulated,the expression of TNF-α,IL-1β and IL-6 mRNA and contents of TNF-α,IL-1β and IL-6 were decreased,the escape latency was shortened,and the time of staying at the original platform quadrant was prolonged in group L + I (P ＜ 0.05).Conclusion LiC1 pretreatment can improve isoflurane-induced cognitive dysfunction and inhibition of inflammatory response in hippocampus is involved in the mechanism in aged rats.

Objective To investigate the effect of melatonin on isoflurane anesthesia-induced cognitive dysfunction in aged rats.Methods Seventy-five male SD rats,aged 18-20 months,weighing 350-400 g,were randomly assigned into 5 groups(n =15 each):control group(group C),1.5％ isoflurane group(group Ⅰ),melatonin 5 mg/kg group(group M1),melatonin 10 mg/kg group(group M2)and melatonin 20 mg/kg group(group M3).Group G inhaled a gas mixture of oxygen and air for4 h and group 1 inhsled 1.5％ isoflurane for4 h.Melatonin 5,10 and 20 mg/kg(in normal saline containing 1％ DMSO)were injected intrsperitoneally at 15 min before anesthesia and 3 h after the beginning of anesthesia in groups M1,M2 and M3 respectively,and then the animals inhaled 1.5％ isoflurane for 4 h.At the end of anesthesia,5 rats in each group were chosen and blood samples were taken to perform arterial blood gas analysis and to detect the blood glucose level and expression of phosphorylated Tau(p-Tau)protein in hippocampus.Ten rats in each group were chosen at 14 d after the end of anesthesia and Morris water maze was performed 3 times a day for 5 consecutive days to assess the cognitive function.Then the animals were sacrificed and hippocampi were removed for detection of p-Tau expression by Western blot.Results There were no significant differences in the parameters of arterial blood gas analysis and blood glucose level among the 5 groups(P ＞ 0.05).Compared with group C,the escape latency at 3-5 d was significantly prolonged,the probe time was significantly shortened,and the expression of p-Tau protein was up-regulated in groups I and M1(P ＜0.05),and no significant change was found in the indexes mentioned above in groups M2 and M3 (P ＞0.05).Compared with groups 1 and M1,the escape latency at 2-5 d was significantly shortened,the probe lime was significantly prolonged,and the expression of p-Tau protein was down-regulated in groups M2 and M3 (P ＜ 0.05).There were no significant differences in the indexes mentioned above between groups I and M1,and between groups M2 and M3(P ＞ 0.05).Conclusion Melatonin(10 and 20 mg/kg)can improve isoflurane anesthesia-induced cognitive dysfunclion in aged rats,which nay be related to inhibition of hyperphosphorylation of Tau protein in hippocampus.