ObjectiveTo determine the depth of sedation induced by target-controlled infusion (TCI) of propofol required to inhibit the swallowing reflex.MethodsThirty-five ASA Ⅰ or Ⅱ healthy volunteers,aged 20-60 yr,with body mass index ＜ 30 kg/m2,were enrolled in the study.Sedation was performed with TCI of propofol.The initial target plasma concentration (Cp) of propofol was set at 2.4 μg/ml and the Cp increased or decreased by 0.2 μg/ml every 5 min until the BIS value reached the predetermined level.The initial Cp of propofol was increased or decreased by 0.3 μg/ml in the next subject according to the BIS value in the previous one.The up-and-down sequential experiment was pedormed.The BIS value was set at 65 in the first subject and the ratio of the BIS value between the two consecutive subjects was 1.1.The BIS value was increased or decreased depending on the occurrence of the swallowing reflex in the previous subject.The half-effective BIS value (BIS50) and 95％confidence interval (CI) of propofol inhibiting the swallowing reflex were calculated by up-and-down sequential method.ResultsThe BIS50 of propofol required to inhibit the swallowing reflex was 64.4 (95％ CI 59.3-69.8).ConclusionThe BIS50 and 95％ CI of propofol administered by TCI and required to inhibit the swallowing reflex is 64,4 (59.3-69.8).

BACKGROUND:Kunming mouse embryonic fibroblasts are the most common feeder layers at present, and there are rare reports addressing C57BL/6 mouse embryonic fibroblasts as feeder layers.
OBJECTIVE:To separate and culture C57BL/6 mouse embryonic fibroblasts in vitro, and produce feeder layers to enlarge the resources of mouse embryonic fibroblasts.
METHODS:C57BL/6 mouse embryonic fibroblasts were isolated and cultured by trypsin digestion method in vitro. The biological characteristics and growth rule of the fibroblasts were investigated, then the feeder layers for the cel culture were produced. The growth of cel colonies on the prepared feeder layer was tested.
RESULTS AND CONCLUSION:C57BL/6 mouse embryonic fibroblasts grew wel with a large amount, by trypsin digestion method at different concentrations. There was no significance in the survival rate after cryopreservation for 1 week, 2 weeks, 1 month, 3 months and 6 months. The cel s were proliferative from the second to fifth passage and declined sharply after the sixth passage. The planted mouse embryonic fibroblasts feeder layers had a high activity within 3 days, but got a sharp decline after 4 days. So it is best to use C57BL/6 mouse embryonic fibroblast feeder layers within 3 days after they’re inactivated. C57BL/6 mouse embryonic fibroblast feeder layer can support embryonic stem cel s and induce pluripotent stem cel s to grow as Kunming mouse embryonic fibroblasts.

Objective To observe the effect of early rehabilitation training combined with continuous passive motion (CPM) on knee joint function after total knee arthroplasty (TKA).Methods23 patients (28 knees) post TKA were randomly divided into the treatment group (15 knees) and control group (13 knees). The patients of the treatment group were treated with early rehabilitation training combined with CPM, and those of the control group were treated with CPM only.ResultsThe scores of knee grade and function grade of the treatment group were not different from those of the control group before treatment ( P>0.05), but two weeks later, there was a significnat difference between two groups ( P<0.01).ConclusionThe early rehabilitation training combined with CPM can improve recovery of function of knee joint after TKA.

Objective To observe and compare the control effect of monitored anesthesia care on stress reaction of operation under local anesthesia.Methods Fourty-five patients who undered ophthalmology and otolaryngology operation and ASA Ⅰ or Ⅱ class,aged 20-55 years were enrolled.They were randomly divided into 3 groups:monitored anesthesia care(M group),general anesthesia (G group),and local anesthesia (L group).All of three groups were performed local nerve block anesthesia.Mean arterial blood pressure (MAP)、heart rate (HR) were recorded at enter room (T1),the strongest operation stimulation (T2),the end of operation (T3).At the same time,the levels of blood glucose,plasma cortisol (Cor) were assayed.The scores of state of anxiety (S-AI) before operation and after operation in two groups were compared.Results The levels of MAP and Cor at T2,T3 and HR at T2 in G group and M group were significantly lower than those at T1:(68.1 ± 8.2),(78.8 ± 12.8) mmHg (1 mmHg =0.133 kPa) vs.(95.7 ± 11.3) mmHg;(80.8 ± 11.7),(86.2 ± 9.0)mmHg vs.(94.7 ± 11.5) mmHg;(207.0 ±71.8),(135.2 ± 60.9) nmol/L vs.(336.7 ± 121.4) nmol/L;(220.8 ± 113.2),(190.4 ± 149.0) nmol/L vs.(347.8 ± 154.6) nmol/L;(68.1 ± 10.6) beats/min vs.(79.9 ± 14.2)beats/min;(70.3 ± 10.1) beats/min vs.(80.6 ± 12.2) beats/min,there were significant differences (P ＜0.05).The level of blood glucose at T3 was significantly lower than that at T1 in G group [(4.9 ± 0.7) mmol/L vs.(5.5 ± 0.6) mmol/L],there was significant difference (P ＜ 0.05).The levels of MAP,Cor at T2 in G group and M group,at T3 in G group were significantly lower than those at the same point in L group:[(68.1 ± 8.2),(80.8 ± 11.7) mmHg vs.(93.4 ± 12.5) mmHg,(207.0 ± 71.8),(220.8± 113.2) nmol/L vs.(367.1 ± 157.3)nmol/L,(78.8 ± 12.8) mmHg vs.(92.6 ± 15.3) mmHg,(135.2 ± 60.7) nmol/L vs.(311.9 ± 165.6) nmol/L],there were significant differences (P ＜ 0.05).The scores of S-AI at postoperative in G group and L group were significantly lower than that at preoperative:(31.5 ± 6.6) scores vs.(44.3 ± 15.0) scores,(35.2 ± 11.5) scores vs.(49.3 ± 14.2) scores,there were significant differences (P ＜ 0.05).Conclusion Monitored anesthesia care can alleviate stress reaction in operation with local anesthesia,while its effect is similar with general nesthesia.

Objective To study the effects of dexmedetomidine on the swallowing reflex. Methods Sixty adult volunteers (ASAⅠ-Ⅱ) were randomly divided into two groups, dexmedetomidine sedation group and propofol sedation group respectively. The BIS value was maintained between 60-65 in both groups. Swallowing reflex was induced by water injection in the pharynx at different depths of sedation, and the swallowing movements were observed. Meanwhile, the RR, SpO2 and OAA/S scores were recorded. Results The swallowing reflex existed in dexmedetomidine sedation group when BIS values was maintained at 60, which had significant difference compared with that in propofol sedation group. The RR, SpO2, OAA/S score in dexmedetomidine sedation group also had statistical difference compared with that in propofol sedation group during swallowing reflex test. Conclusions Slow infusion of dexmedetomidine to maintain the BIS value above 60 will not affect swallowing reflex, and can obtain satisfactory hemodynamic stability. It′clinical safety is high.

AIM: To investigate the effects of caveolin-1 (Cav-1) scaffolding domain peptide, cavtratin, on lipopolysaccharide (LPS)-induced mouse acute lung injury and heme oxygenase-1 (HO-1) activity.METHODS: Adult male BALB/c mice were randomly divided into 6 groups (n=8 to 10): control, Antennapedia internalization sequence (AP), LPS, LPS+hemin, LPS+ hemin+cavtratin and LPS+hemin+cavtratin+zinc protoporphyrin IX (ZnPP) groups.After LPS administration for 24 h, the lung pathological changes, the wet/dry weight (W/D) ratio of lung tissues, total cell number in bronchoalveolar lavage fluid and serum lactate dehydrogenase activity were measured.The co-localization of HO-1 and Cav-1 was displayed by immunofluorescence, and the HO-1 activity were detected.The mRNA expression of pro-inflammatory cytokines IL-1β, IL-6, TNF-α, MCP-1 and iNOS was detected by real-time PCR.RESULTS: The mice in LPS+hemin+cavtratin group had the decreased interaction between HO-1 and Cav-1, and the increased HO-1 activity compare with LPS group (P<0.05).Compared with LPS group, the pulmonary damage was attenuated in LPS+hemin+cavtratin group, and the injury indexes, including W/D ratio, total cell number in bronchoalveolar lavage fluid and lactate dehydrogenase activity in the serum, and the mRNA expression of inflammatory cytokines all decreased (P<0.05).HO-1 activity inhibitor ZnPP abolished the above protective effect of cavtratin on the lung tissues with LPS-induced acute lung injury.CONCLUSION: Cavtratin has beneficial effects on the lung with LPS-induced acute injury by restoring the HO-1 activity.

Objective To evaluate the value of whole genome amplification (WGA) combined with array comparative genomic hybridization (aCGH) in prenatal diagnosis. Methods Array CGH were performed by the DNA of 18 prenatal specimens , which were amplified by WGA because of the low DNA yield. Result 3 of the 18 fetuses were 45, X0 and 9 of 15 fetuses with normal aCGH results showed healthy outcome. Conclusion It’ s feasible for prenatal diagnosis using WGA combined with aCGH which not only can shorten the reporting time but also keep the sensitivity and accuracy of detection.

AIM:To study the effects of methylene blue (MB) on myocardial ischemia/reperfusion (I/R)-induced mitochondrial injury in isolated rat hearts.METHODS:Spragure-Dawley (SD) rats were divided into 3 groups randomly (n=6): control group, I/R model group and MB treatment group (IR+MB group).The isolated rat hearts were prepared and set up to Langendorff perfusion.The rats in I/R+MB group received MB (2 mg/kg) by intraperitoneal injection 2 h before operation.The hearts in control group were perfused with K-H solution for 110 min consecutively.The hearts in I/R group and I/R+MB group were in equilibrium for 20 min, following by 45 min of global ischemia, and then reperfused for 60 min.The heart rate (HR), left ventricular developed pressure (LVDP), left ventricular pressure maximum change rate (±dp/dtmax) and left ventricular end-diastolic pressure (LVEDP) were recorded.The perfusate was collected to determine the activity of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH).The contents of reactive oxygen species (ROS), malondialdehyde (MDA) and adenosine triphosphate (ATP), and the activity of superoxide dismutase (SOD) in the myocardial tissues were all determined.Histopathological examination of left ventricle was performed.The mitochondria from the heart tissues was isolated and the mitochondrial swelling and mitochondrial membrane potential (MMP) were analyzed.RESULTS:Compared with control group, the hearts in I/R group showed poorer function, higher CK-MB and LDH levels in the perfusate, increased ROS and MDA contents, higher SOD activity and less ATP content in the heart tissues (P<0.05).Furthermore, the mitochondrial swelling level increased and MMP reduced in I/R group (P<0.05).Compared to I/R group, MB improved heart function and reduced the release of CK-MB and LDH (P<0.05).MB also decreased ROS and MDA contents, and increased the activity of SOD and the content of ATP (P<0.05).In addition, MB alleviated mitochondrial swelling and restored the reduced MMP (P<0.05).CONCLUSION: MB protects the isolated rat hearts from I/R-induced injury by attenuating the damage of mitochondria.

Objective The aim of this study was to investigate the polymorphism of SLA class II genes in Canadian SPF Yorkshire and Landrace pigs.Methods Blood samples were obtained from 15 SPF Yorkshire and 22 Landrace pigs for isolation of peripheral blood mononuclear cells respectively, and the DQB1, DRB1 and DQA genes were amplified by PCR after reverse transcription.SLA class II genes were obtained by analyzing the direct and cloning result.The polymorphism of alleles was analyzed using the DNAsp 5.0 software.Results A total of 25 alleles were identified at three genes, including eight DQB1, ten DRB1 and seven DQA, and three alleles were submitted the complete sequences for the first time.The official allele names were assigned as SLA-DQB1*0212 (KU754590), SLA-DQB1*0203 (KU754591) and DRB1*06:07(KU754601) by the SLA Nomenclature Committee.Three novel DQA alleles were discovered.Five of the 15 amino acids, one of the 16 amino acids and 11 of the 19 amino acids, which bind processing antigens, showed well conserved among the alleles of DQB1, DRB1 and DQA genes in the SPF Yorkshire and Landrace pigs, respectively.Neighbor-joining tree showed that the three genes were divided into two clusters, respectively.There was a close relationship between SPF Yorkshire and Landrace pigs and foreign Yucatan miniature pigs, and it showed no obvious genetic distance with other pigs.Conclusions A total of 25 SLA class II alleles have been identified successfully in this study, and there are more abundant polymorphism for them.There is a widely distribution for SLA class II alleles identified in this study in other pig breeds.It is critical for the eventual future use of SPF Yorkshire and Landrace pigs as classical laboratory animal models.

Objective To establish effective method for large-scale purification of islet cells from pig pan-cress. Methods Pig pancreas tissue was digested with collagenase P followed by purification in a HCA-Fi-coil dis continuous gradient using Cobe2991 cell separator. After isolation, the islet cell yield and purity were evaluated with light microscope with DTZ staining, and the islet function assessed by insulin release as-say in vitro. Results The number of the islets coll ected from each pancreas averaged (275 000±20 895)islet equivalents (IEQ) before purification, and (230 350±26 679) IEQ after the purification with discon-tinuous gradient centrifugation. From each gram of the pancreatic tissue, (2710±229) IEQ were obtained with an average purity of (50.2±1.95) %. The purified islets responded well to high-concentration (16.7 mmol/L) glucose stimulation with a 4. 74-fold increase of insulin secretion over the basal level (3.3 mmol/L, P <0.001). Conclusion The established method can be applicable for large-scale purifi-cation of fully functional islet cells from pig pancreas.