Objective To determine the median effective effect-site concentration (EC50 ) of remifentanil inhibiting responses to laryngeal mask airway insertion when combined with propofol in elderly male patients . Methods Thirty ASA physical status Ⅰ or Ⅱ male patients ,aged 65>yr ,with body mass index <30 kg/m2 , scheduled for elective transurethral resection of bladder tumor or prostate under general anesthesia ,were enrolled in this study .Anesthesia was induced with target-controlled infusion of propofol with a target plasma concentration (Cp) of 3 μg/ml .When Observer′s Assessment of Alertness/Sedation (OAA/S ) score ≤1 ,remifentanil target-controlled infusion was started with the initial target Cp set at 4.0 ng/ml . The concentration of propofol was adjusted until BIS value reached 55-65 ,and then the laryngeal mask airway was inserted .Modified Dixon’s up-and-down method was used to determine the Cp of remifentanil . Each time the Cp of remifentanil increased/decreased in the next patient depending on whether or not the response to laryngeal mask airway insertion occurred . The ratio of the two successive Cps was 1.2 .The response to laryngeal mask airway insertion was defined as development of coughing ,laryngospasm and/or body movement during insertion or within 3 min after insertion .The number of patients in whom inhibition of responses to insertion was effective/ineffective was recorded .The EC50 of remifentanil required to inhibit responses to laryngeal mask airway insertion and the 95% confidence interval when combined with propofol were calculated .Results The EC50 (95% confidence interval ) of remifentanil required to inhibit responses to laryngeal mask airway insertion was 1.86 (1.64-2.12) ng/ml when combined with propofol in elderly male patients .Conclusion The EC50 of remifentanil required to inhibit responses to laryngeal mask airway insertion is 1.86 ng/ml when combined with propofol in elderly male patients .

Objective Comparing the effects of ultrasound with nerve stimulation guided obtu-rator nerve block(ONB)with simple nerve stimulation guided ONB for transurethral resection of blad-der tumor(TURBT),so as to realize the validity and advantages of ultrasound with nerve stimulation guided ONB.Methods Sixty ASA Ⅰ or Ⅱ,male or female,age 39-77 years old,BMI 1 9-30 kg/m2 patients undergoing elective TURBT were randomly divided into two groups,nerve stimulation group (group S)and ultrasound and nerve stimulation group (group US).Success rate of the first puncture, visual analog scale (VAS)pain score,insertion-adductor contraction interval (ICI),puncture times corresponding to ICI,adductor strength,incidence of complications and validity were observed during and after ONB.Results There was no significant difference of the general validity,adductor strength and complication incidence between the two groups.The success rate of the first puncture was signifi-cantly higher in group US than that in group S (P < 0.01).VAS pain score,ICI and puncture times were significantly lower in group US than those in group S (P < 0.05).Conclusion Compared with simple nerve stimulation guided ONB,ultrasound with nerve stimulation guidance showed less punc-ture time,more accurate positioning and more comfort.

Objective To evaluate development of gastro-esophageal reflux (GER) during laparoscopic surgery in lateral jack-knife position under general anesthesia through comparing with reverse Trendelenburg/ Trendelenburg position in the patients lying supine.Methods Ninety patients of both sexes,aged 18-64 yr,of ASA physical status Ⅰ or Ⅱ,with body mass index of 18-30 kg/m2,scheduled for elective laparoscopic surgery under general anesthesia,were randomly divided into 3 groups (n =30 each):lateral jack-knife position group (group L),Trendelenburg position group (group T) and reverse Trendelenburg position group (group Tre).Anesthesia was induced with midazolam,sufentanil,propofol and cisatracurium besylate and maintained with propofol and remifentanil given by target-controlled infusion.A pH-sensitive probe was inserted through nose into the lower esophagus and pH value was continuously recorded until 1 min after extubation.GER was defined as pH value ≤ 4 lasting for ≥ 1 min in the lower esophagus during surgery.The development of GER during surgery and the lowest pH value in the lower esophagus when GER developed were recorded.Results Compared with group Tre,the incidence of GER (27％) and total number of times GER had occurred were significantly increased in group L,and no significant changes were found in the indices mentioned above in L and T groups.When GER developed,the lowest pH value in the lower esophagus was 2.1 ± 1.3,2.6 ± 1.2 and 3.5 in L,T and Tre groups,respectively.Conclusion The incidence of GER is 27 ％ during laparoscopic surgery when the patients are in lateral jack-knife position and it is higher than that obtained with reverse Trendelenburg position in the patients lying supine.

Objective To investigate the effect of L-carnitine on the apoptosis in Schwann cells induced by high glucose.Methods The cell line RSC96 cultured in vitro were seeded in 96-well plates at a density of 1.5 × 104/ml (200 μl/well) or in 6-well plates at a density of 2 × 105/ml (2 ml/well) and cultured for 24 h.The cells were randomly divided into 4 groups (n =24 each) using a random number table:normal control group (group C),high glucose group (group H),high glucose + L-carnitine group (group H + L),and mannitol osmotic control group (group M).The cells in group C were incubated in the plain culture medium containing normal glucose (5.6 mmol/L).The cells were incubated in the medium containing glucose 50 mmol/L in group H or in the medium containing glucose 50 mmol/L and L-carnitine 50 μmol/L (final concentration) in group H + L.The cells were incubated in the medium containing normal glucose (5.6 mtmol/L) and mannitol 44.4 mmol/L in group M.At 48 h of incubation,cell growth conditions were observed under inverted microscope,superoxide dismutase (SOD) activity was measured by xanthine oxidase method,malondialdehyde (MDA) content was measured by thiobarbituric acid test,cell viability was measured by MTT assay and cell apoptosis was measured by flow cytometry.The expression of activated caspase-3 and poly (ADP-ribose) polymerase-1 (PARP-1) protein was detected by Western blot.Results Compared with group C,the cell viability and SOD activity were significantly decreased,MDA content and apoptotic rate were increased,and the expression of activated caspase-3 and PARP-1 protein was up-regulated in H and H + L groups,and no significant changes were found in group M.Compared with group H,the cell viability and SOD activity were significantly increased,MDA content and apoptotic rate were decreased,and the expression of activated caspase-3 and PARP-1 protein was down-regulated in group H + L.Conclusion L-camitine can attenuate high glucose-induced apoptosis in Schwann cells by inhibiting oxidative stress responses and down-regulating the expression of activated caspase-3 and PARP-1.

Objective To determine the effective volume of 1.5％ lidocaine for obturator nerve block (ONB) in 50％ of patients (EV50) undergoing transurethral resection of bladder tumor (TURBT).Methods Thirty-six ASA physical status Ⅰ or Ⅱ patients with bladder tumor,aged 18-64 yr,with body mass index of 19-30 kg/m2,scheduled for elective TURBT and required ONB before TURBT,were enrolled in the study.ONB was performed with 1.5 ％ lidocaine using the pubic approach under the guidance of a nerve stimulator.The volume of 1.5％ lidocaine was determined by up-and-down sequential trial.The initial volume of hdocaine was 10 ml and the ratio between the two successive volumes was 1.1.Successful ONB was considered to be positive response.The EV50 and 95 ％ confidence interval (CI) of 1.5 ％ lidocaine for ONB were calculated.Results The EV50 of 1.5 ％ lidocaine for ONB was 5.53 rnl and the 95 ％ CI was 5.10-6.00 ml.Conclusion The EV50 of 1.5 ％ lidocaine is 5.53 ml when used for ONB in patients undergoing TURBT.

Glucose-6-phosphate dehydrogenase (G6PD) derives from the expression of the house-keeping gene G6PD. Recent studies have indicated that G6PD is related to tumor genesis, growth, clinical phenotype, therapy, and prognosis. To elucidate the relationship between G6PD and cancer, three siRNA sequences and one negative control sequence were designed based on the 3' noncoding region of the human G6PD gene. Two complementary single-strand DNA (sense and antisense) were designed and synthesized based on siRNA sequences. The DNA fragments were annealed and ligated to the GFP expression vector pRNAT-U6.2/Lenti. One siRNA with higher interference efficiency than the other two was found after siRNA plasmid transfecting human skin A375 melanoma cells. After lentivirus particle packaging and virus production, the A375 cells were infected, and the single cell clone was acquired and cultured to establish the stable cell strain. Western blotting showed that the endogenous G6PD in the stable A375 cell strain was 0.257 ? 0.074, which was 11.17% of G6PD expression (2.301 ? 0.286) in wild type A375 cells. The final siRNA interference efficiency in this stable cell strain was 88.83%. The G6PD activity of A375-G6PD?驻 was 21.53% of A375-WT. Further study showed that A375-G6PD△ doubling generation time prolonged, and its proliferation was greatly inhibited and the cloning efficiency lowered 25%(P

Objective To determine the median effective concentration (EC50) of lidocaine for obturator nerve block (ONB) guided by a nerve stimulator in patients undergoing transurethral resection of bladder tumor (TURBT).Methods American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients with bladder tumor,scheduled for elective TURBT,required ONB according to the results of cystoscopy or CT examination performed before operation,with body mass index of 19-30 kg/m2,aged 18-64 yr,were enrolled in the study.ONB was performed with lidocaine using the suprainguinal approach under the guidance of a nerve stimulator.The concentration of lidocaine was determined by up-and-down sequential trial.The initial concentration of lidocaine was 1.5％,and the ratio between the two successive concentrations was 1.2.Successful ONB was considered to be positive response.The EC50 and 95％ confidence interval of lidocaine for ONB guided by a nerve stimulator was calculated.Results The EC50 of lidocaine was 0.57％,and the 95％ confidence interval was 0.55％-0.59％ when used for ONB guided by a nerve stimulator.Conclusion The EC50 of lidocaine is 0.57％ when used for ONB guided by a nerve stimulator in the patients undergoing TURBT.

Objective To compare the efficacy of suprainguinal approach and pubic tubercle approach to obturator nerve block (ONB) in patients undergoing transurethral resection of bladder tumor.Methods Sixty ASA physical status Ⅰ or Ⅱ patients of both sexes,aged 41-80 yr,with body mass index of 17.5-31.0 kg/m2,scheduled for elective transurethral resection of bladder tumor,were randomly divided into 2 groups (n =30 each) using a random number table:pubic tubercle approach group (group P) and suprainguinal approach group (group S).Nerve blocks were performed using a 100-mm insulated needle for ONB (21-gauge) under the guidance of a nerve stimulator.In group P,the insertion point of the needle was 1.5 cm lateral and 1.5 cm inferior to the pubic tubercle.In S group,the insertion point of the needle was at the midpoint of the line drawn in the inguinal crease between the femoral artery and the inner border of the adductor longus tendon and the needle was advanced 3 cm cephalad in the major axis of thigh.The number of puncture eliciting contraction of adductor muscle,time taken to elicit contraction of adductor muscle starting from onset of puncture,depth of puncture,and highest visual analog scale (VAS) pain scores during application of the block were recorded.The myodynamia of adductor muscle was evaluated.The development of complications was also recorded.Results Compared with group P,the number of puncture,highest VAS scores,and myodynamia of adductor muscle at 4 and 6 min of blockade were significantly decreased,the time taken to elicit contraction of adductor muscle was shortened,and the success rate of puncture at first attempt was increased in group S (P ＜ 0.05 or 0.01).There was no significant difference in the incidence of puncture point bleedings between the two groups (P ＞ 0.05).Conclusion The suprainguinal approach for ONB offers more accurate location,faster onset,lighter degree of noxious stimulation and better safety than the pubic tubercle approach in patients undergoing transurethral resection of bladder tumor.

BACKGROUND:Recent studies have found that stem cellpluripotency and differentiation is regulated by many long non-coding RNAs (LncRNAs). The expression and effect of LncRNA AK089560 during differentiation of stem cells is unclear. OBJECTIVE:To investigate the expression of LncRNA AK089560 in mesenchymal stem cells C3H10T1/2 undergoing osteogenic and adipogenic differentiation. METHODS:Osteogenic differentiation of mesenchymal stem cells C3H10T1/2 was induced by recombinant human bone morphogenetic protein-2 and evaluated using alkaline phosphatase staining. The adipogenic differentiation of mesenchymal stem cells C3H10T1/2 was induced by three factors (dexamethasone, indomethacin and insulin) and evaluated by oil red O staining. The dynamical expression of LncRNA AK089560 was detected by qRT-PCR assay. The AK089560 secondary structure was predicted using RNAfold software. The relationship between AK089560 and neighboring protein-coding genes was analyzed using UCSC genome browser and visualized by fancyGENE online software. RESULTS AND CONCLUSION:Over 70%of C3H10T1/2 cells were positive for alkaline phosphatase after osteogenic induction and more than 80%of the cells positive for oil red O staining after adipogenic induction. qRT-PCR results showed that the expression of LncRNA AK089560 at days 2, 4, 6 of both osteogenic and adipogenic differentiation was significantly decreased compared with the control group (P<0.05). Bioinformatics analysis showed that there was a stem-loop structure for AK089560 and sense overlap relationship between AK089560 and protein-encoding gene Sema3a. These findings indicate that LncRNA AK089560 expression is reduced during osteogenic differentiation and adipogenic differentiation, showing that AK089560 may be involved in regulating the multi-directional differentiation of stem cells.

Objective To investigate the effects of ketamine combined with moderate hypothermia on brain ischemia-reperfusion (I/R) injury in a rat model of asphyxial cardiac arrest. Methods Fifty healthy Wistar rats of both sexes aged 4.0-4.5 months, weighing 410-510 g were randomly allocated into 5 groups (n = 10each): group Ⅰ sham operation (group S), group Ⅱ asphyxial cardiac arrest (group ACA), group Ⅲ ketamine (group K), group Ⅳ moderate hypothermia (group MH) and group Ⅴ K + MH. The animals were anesthetized with intraperitoneal (IP) phenobarbital 20 mg/100 g, tracheostomized and mechanically ventilated (RR 60 bpm,FiO2 50%), PaCO2 was maintained at 35-45 mm Hg. Cardiac arrest was induced by clamping tracheal tube until ECG activity disappeared and MAP ＜ 15 mm Hg. Resuscitated was started 5 min later. MAP ＞ 60 mm Hg and HR ＞ 250 bpm were considered to be signs of successful resuscitation. Dead animals and animals in which resuscitation time was longer than 5 min were excluded from the study. In group K ketamine 100 mg/kg was administered IP at 5 min before asphyxia. In group MH hypothermia was started as soon as asphyxia was started and body temperature was maintained at 30-35 ℃. After successful resuscitation, the animals were sacrificed. Their brains were removed for determination of brain water content and p-caspase-3 expression in hippocampus. Results Brain I/Rsignificantly increased brain water content and p-caspase-3 expression in group ACA. MH alone significantly attenuated 1/R-induced brain edema and decreased p-caspase-3 expression, while ketamine alone only significantly decreased p-caspase-3 expression but did not decrease I/R-induced brain edema. MH + K decreased p-caspase-3expression further but did not reduce brain edema further as compared with MH alone. Conclusion Ketamine combined with moderate hypothermia provides better protection against brain I/R injury.