Objective To evaluate the effect of intrathecal (IT) NR2B antisense oligonucleotide (aNR2B) on cognitive function in morphine-dependent rats.Methods Male SD rats weighing 230-270 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital 60 mg/kg.IT catheter was placed at L3-4 interspace according to the technique described by Yang. Thirty rats in which IT catheter was successfully placed without any complication were randomly divided into 3 groups(n=10 each):control group (group C), morphine dependence group (group MD) and group aNR2B.Morphine dependence was induced in group MD and aNR2B by increasing doses of morphine for 6 days. The initial dose of morphine was 10mg/kg injected subcutaneously (SC) twice a day and was increased by 10 mg/kg.every other day.The final dose was 50mg/kg. Then morphine 30 mg/kg was administered SC once a day for 4 weeks. aNR2B 15 nmol was administered IT at 30 min before SC morphine every day in group aNR2B.In control group normal saline was administered instead of morphine. Morris water maze was used to assess the cognitive function at 0 (T0, baseline),1 and 3 weeks of morphine administration (T1,T2).The escape latency and the number of times the animals crossing the plateform were recorded. The animals were killed after the test and the hippocampus was isolated for determination of choline acetytransferase(ChAT)expression.Results There was no significant difference in the baseline escape latency and the baseline number of times the animals crossing the plateform at T0 among the 3 groups. The escape latency was significantly prolonged and the number of times the animals crossing the plateform decreased at T1 and T2 as compared with the baseline at T0 in group MD.The ChAT expression was significantly down-regulated in group MD as compared with control group. IT aNR2B significantly ameliorated cognitive dysfunction at T1 and T2 and increased ChAT expression in group aNR2B compared with group MD.Conclusion IT NR2B antisense oligonucleotide can attenuate cognitive dysfunction through up-regulation of ChAT expression in hippocampus in morphine-dependent rats.

Objective To investigate the effect of NR2B antisense oligonucleotide on naloxone-induced withdrawal responses in morphine-dependent rats. Methods Famale SD rats weighing 230-270 g were anesthetized with intraperitoneal pentobarhital 60 mg/kg. Intrathecal (IT)catheter was placed at L3,4 interspace.Thirty-two rats in which FT catheter was successfully placed were randomly divided into 4 groups ( n = 8 each) : group C control; group MD morphine dependence; group AO NR2B antisense oligonucleotide (aNR2B) and group SO NR2B sense oligonucleotide (sNR2B) . In group MD, AO, SO chronic morphine dependence was induced by increasing doses of subcutaneous morphine for 6 days. The initial dose of morphine was 10 mg/kg twice a day and was increased by 10 mg/kg twice every other day and reached 50 mg/kg on the 6th day. In group AO and SO IT aNR2B or sNR2B 15 nmol was administered simultaneously with subcutaneous morphine. Morphine withdrawal responses was induced by IT naloxone 4 mg/kg and scored based on the responses (0 = normal; higher scores signify severer responses) . The weight loss was calculated.The expression of NR1, NR2A and NR2B mRNA in hippocampus was determined by RT-PCR. Results The morphine withdrawal syndrome and weight loss were significantly incresed in group MD, AO and SO, while NR2B mRNA expression in hippocampus was up-regulated in group MD and SO compared with group C. The morphine withdrawal syndrome and weight loss were significantly decreased, NR2A mRNA expression in hippocampus was up-regulated and NR2B mRNA expression was down-regulated in group AO compared with group MD. There was no significant difference in NR1 mRNA expression between the 4 groups . Conclusion NR2B antisense oligonucleotide can suppress morphine withdrawal responses through the regulation of NMDA receptor level and construction in hippocampus.

Objective To analyze the characteristics and clinical value of time intensity curve (TIC) of contrast-enhanced ultrasound (CEUS) in recurrent small hepatocellular carcinoma (RSHCC) and primary small hepatocellular carrcinoma (PSHCC). Methods Sixty-five cases of RSHCC (all lesions ≤3 cm) were devided into group B1 with 42 cases of RSHCC (≤2 years ) , and group B2 with 23 cases of RSHCC ( > 2 yeras ) and group A invloved 49 cases of PSHCC (all lesions ≤3 cm). Enhancement patterns in arterial, portal and delayed phase were evaluated respectively in three groups through CEUS and analytic software Sonoliver was applied to obtain quantitative features of CEUS in the region of interest. Receiver operating characteristic (ROC) was drawn and the area under curve (AUC) was calculated. Results CEUS showed hyper-enhancement difference in arte-rial phase in group B2 (72.4%) and group A (94.8%)(P′ = 0.008) showed statistical significance, but no sig-nificance was found in enhanced iso in portal phase (P = 0.078). Hypo-enhancement in the delayed phase in group B2 (75.9%), group A (96.6%) and group B1 (95.3%) (P′ = 0.003, P′ = 0.005). TIC showed HT difference (half time of descending) in B2 group, A group and B1 group (P′ = 0.007, P′ = 0.013) indicated statistical significance but RT, TTP, MTT(P = 0.319,P = 0.104, P = 0.461) showed no difference. AUC was 0.841 (half time of descending). Conclusions Enhancement patterns of CEUS (RSHCC) are related to recur-rent time . En hancement patterns of RSHCC (> 2 years ) is not typical so CEUS should be combined with quanti-tative analysis of TIC to provide reference for its treatment and prognosis.

Objective To identify,construct and express scFv CD133,verify its biological function.Methods VL and VH were isolated from hybridoma of mAb CD133 by using antibody engineering technology.Its DNA sequencing and CDR were determined.scFv CD133 was then cloned into pET32a,transformed into Origami,induced by IPTG,purified by Ni2+-NTA His resin.Its affinity and specificity were tested by NH4SCN elution and ELISA.Results The size of VL and VH of scFv CD133 was 339 bp and 342 bp,which coded 113 and 114 amino acid separately.Its VL belonged to mouse Igκ chain and VH belonged to mouse IgG heavy chain subtype I.The molecular weight of scFv CD133 was about 27 × 103 which was testified by SDSPAGE and Western blot.Its affinity and specificity were also verified.Conclusion scFv CD133 has been successfully constructed and expressed in Origami,which could supply basis for target therapy of CD+133 cancer stem cell.

Objective To investigate the value of dynamic contrast?enhanced MRI (DCE?MRI) in the differential diagnosis of glioblastoma and brain metastases. Methods Twenty patients with high grade gliomas and 20 cases patients with brain metastases proved by surgery and pathology were collected, and patients were examined with conventional MRI and DCE?MRI preoperatively. The ROIs were manually placed in solid parts of the tumors and their surrounding tissues to calculate Ktrans, Kep and Ve values. The Ktrans, Kep and Ve values differences for the solid part and surrounding tissues of the two brain tumors were compared by two independent sample t test. The correlation between Ktrans of the solid parts of the two brain tumors and Ktrans, Kep and Ve values of their surrounding tissues were studied by Pearson correlation analysis. Results The Ktrans, Kep and Ve values of glioblastoma were(0.258 ± 0.063)min-1,(0.398 ± 0.082)min-1, 0.632±0.084, the Ktrans, Kep and Ve values of brain metastases were(0.233±0.053)min-1,(0.357±0.042)min-1, 0.672±0.113. There were no significant differences between the glioblastoma and brain metastases for Ktrans, Kep and Ve values(t=-1.354,-1.982, 1.276, all P>0.05). The Ktrans, Kep and Ve values of surrounding tissues of glioblastoma were(0.093±0.032)min-1,(0.411±0.089)min-1, 0.107±0.021, the Ktrans, Kep and Ve values of surrounding tissues of brain metastases were(0.033±0.010)min-1,(0.204±0.045)min-1, 0.069±0.017. The Ktrans, Kep and Ve values of surrounding tissues between glioblastoma and brain metastases had significant difference (t=-7.978,-9.303,-6.203, all P<0.05). The Ktrans of glioblastoma were correlated with Ktrans, Kep and Ve values of their surrounding tissues (r=0.759, 0.464, 0.651, all P<0.05); The Ktrans values of brain metastases had no relationship with Ktrans, Kep and Ve values of their surrounding tissues (P>0.05). Conclusion The DCE?MRI can quantitatively display the microvascular permeability and accurately evaluate the damage of blood?brain barrier of glioblastoma and brain metastases, which has an important value in studying biological characteristics and differential diagnosis of the two brain tumors.

Objective To evaluate the efficacy of ultrasound-guided continuous transverses abdominis plane (TAP) block when used for postoperative analgesia in the patients undergoing total hysterectomy.Methods Forty American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients,aged 38-64 yr,weighing 50-80 kg,undergoing elective total hysterectomy with general anesthesia,were divided into 2 groups using a random number table:continuous TAP block group (CTAP group,n =21) and patient-controlled intravenous analgesia (PCIA) group (n=19).In group CTAP,bilateral TAP block was performed with 0.2％ ropivacaine 20 ml under ultrasound guidance before operation,and 0.2％ ropivacaine 5 ml/h was infused into bilateral TAPs after extubation.In group PCIA,the patients received PCIA with sufentanil 1 μg/ml after extubation,and the PCIA pump was set up to deliver a 2 ml bolus dose,a 15 min lockout interval and background infusion at a rate of 2 ml/h.Analgesia lasted until 72 h after operation in both groups.When visual analog scale＞4,morphine 5 mg was intramuscularly injected as rescue analgesic.The recovery time of postoperative intestinal function,length of hospital stay,patient's satisfaction with analgesia,requirement for rescue analgesia,TAP block-related adverse reactions and development of postoperative nausea and vomiting were recorded.In group CTAP,blood samples were collected from the peripheral vein immediately after the end of operation and at 2,6,12,24,48 and 72 h after operation for determination of concentrations of ropivacaine in plasma and free ropivacaine in plasma using high-performance liquid chromatography.Results Compared with group PCIA,the requirement for rescue analgesia and incidence of nausea and vomiting were significantly decreased,the recovery time of postoperative intestinal function was shortened,the score for patient's satisfaction with analgesia was increased (P＜0.05),and no significant change was found in the length of hospital stay in group CTAP (P＞0.05).No TAP block-related adverse reactions were found in group CTAP.In group CTAP,the concentration of ropivacaine in plasma began to increase at 2 h after operation and peaked at 48 h after operation,the concentration of free ropivacaine in plasma began to increase at 2 h after operation and peaked at 24 h after operation (P＜0.05).Conclusion Ultrasound-guided continuous TAP block produces good analgesic efficacy when used for the patients undergoing total hysterectomy.

Objective To compare two different regimens of ultrasound-guided Continuous ad-ductor canal block (CACB)for postoperative analgesia and early ambulation after total knee arthro-plasty (TKA).Methods Sixty-seven patients scheduled for unilateral TKA undergoing spinal anes-thesia,13 males and 54 females,aged 18-85 years,BMI 18-30 kg/m2,ASA physical status Ⅰ-Ⅲ, were randomly divided into the continuous infusion group A (n=34)and the intermittent boluses group B (n=33).After the operations,ultrasound-guided CACB were administered and 20 ml of 0.2% ropivacaine was given as the loading dose.From then on,patients in both groups used electronic analgesic pumps containing 240 ml of 0.2% ropivacaine for postoperative analgesia.5 ml/h of 0.2% ropivacaine was continuously infused for 48 hours in the group A.5 ml of 0.2% ropivacaine was automated injected every 60 minutes in the group B.All infusion pumps were setted at a bolus dose of 5 ml,with a lock time of 30 minutes.The total consumptions of analgestic pum solution and dezoine, quadriceps muscle strength, active range of knee flexion, ambulation distance and occurrences of adverse reactions such as nausea and vomiting,dizziness,drowsiness,extravasating and errhysis were recorded at different time points postoperatively.Results The total consumptions of analgestic pum solution at 12,24 h postoperatively of group B were significantly reduced than that of group A (P<0.05).The 48 h total dezoine consumption of group B was significantly reduced than group A (P<0.05).Active range of knee flexion at 24,48 h and ambulation distance at 48,72 h of group B were significantly higher than group A (P<0.05).There was no statistical difference in quadriceps muscle strength between group A and group B.The incidence of nausea and vomiting in group A was significantly higher than that in group B,and there were no statistical difference in other adverse reactions between group A and group B.Conclusion Compared with the continuous infusion group,the intermittent bolus group for CACB after TKA can provide better analgesic effect and de-crease opioid use postoperatively,with little effect on motor nerve,promoting early ambulation.

Objective To evaluate the effect of obstructive jaundice on the accuracy of left ventricular end-diastolic volume (LVEDV) and stroke volume variability (SVV) in monitoring fluid responsiveness.Methods Thirty patients of both sexes,aged 45-60 yr,weighing 55-70 kg,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,with New York Heart Association Ⅰ,scheduled for elective pancreatoduodenectomy,were divided into 2 groups according to the serum total bilirubin levels:A group (serum total bilirubin ≥ 17 μmmol/L,n =16) and B group (serum total bilirubin＜ 17 μmmol/L,n =14).Six percent hydroxyethyl starch 500 ml was infused over 40 min after anesthesia induction.The parameters of VigileoTM such as cardiac output (CO),SVV,systemic vascular resistance (SVR) and pulmonary capillary wedge pressure and indices measured by transesophageal three-dimensional echocardiography such as LVEDV,left ventricular end-systolic volume,CO',left ventricular ejection fraction (LVEF) and ratio of mitral peak velocity of early filling (E) to early diastolic mitral annular velocity (e',E/e'ratio) were recorded before and after fluid loading.Results Compared with that before fluid loading,SVV was significantly decreased in two groups,and CO,LVEDV,CO'and LVEF were significantly increased in group B,and E/e'ratio was significantly increased in group A (P＜0.05).Compared with group B,CO,SVR,CO'and LVEF were significantly decreased,and pulmonary capillary wedge pressure was increased in group A (P＜0.05).Conclusion Obstructive jaundice causes decrease in the accuracy of LVEDV in monitoring fluid responsiveness and no effect on SVV.

Objective:To evaluate the influence of goal-directed volume management based on cardiac output index (CI), intrathoracic blood volume index (ITBVI) and extravascular lung water index (EVLWI) in patients undergoing off-pump coronary artery bypass surgery.Methods:Forty patients (ASA 2 to 3 grade) undergoing off-pump coronary artery bypass surgery in Lanzhou University Second Hospital from January 2017 to December 2018 were selected. The patients were divided into 2 groups by random digits table method with 20 cases in each group: study group (goal-directed fluid therapy treatment with CI, ITBVI and EVLWI) and control group (conventional fluid therapy). The control group was given central venous pressure (CVP) monitoring rehydration, and the study group was given PiCCO hemodynamic monitoring indicators. The CVP, CI, ITBVI and EVLWI for fluid management were measured. Accurate assessment of volume status of patients was done. The study group received goal-directed fluid therapy based on CVP, CI, ITBVI and EVLWI, with the goal of CI in the 3.0 to 5.0 L/(min·m 2) range, ITBVI in the 800 to 1 000 ml/m 2 range and EVLWI in the 3.0 to 7.0 ml/kg range. The heart rate, mean arterial pressure (MAP), urine volume, central venous oxygen saturation (ScvO 2), lactic acid and renal function were monitored. The ventilator withdrawal time, hospitalization in ICU, length of stay, incidence of acute pulmonary edema, incidence of acute renal failure, mortality of 30 d after surgery were recorded and compared between the two groups. Results:Tissue perfusion and urine volume of the study group was significantly improved compared with that of control group ( P<0.05). ScvO 2 of the study group was higher than that of the routine group ( P<0.05). The concentration of lactic acid of the study group was lower than that of the routine group ( P<0.05). The incidences of acute pulmonary edema, acute renal insufficiency and mortality of the study group were lower than those of the routine group (5.0% vs. 15.0%, 5.0% vs. 10.0% and 5.0% vs. 15.0%), and there were statistical differences ( P<0.05). The length of stay and hospitalization in ICU were both lower than those in the control group ( P<0.01). Conclusions:Goal-directed fluid therapy based on CI, ITBVI and EVLWI can effectively optimize the cardiac preload of patients undergoing off-pump coronary artery bypass surgery, improve cardiac output, ensure microcirculation perfusion, maintain the balance of oxygen supply and demand, and reduce the incidence of complications and mortality.

Objective:To investigate the mechanism of cathepsin G(CatG) in improving the treatment efficacy of T cells in gliomas.Methods:(1) Clinical data of 397 glioma patients in the glioma database were collected, Kaplan-Meier method was used to perform survival analysis, and the correlation between CTSG and β2-microglobulin ( β2M) mRNA expressions in glioma tissues was analyzed. (2) Glioma stem cell (GSC) 387 and GSC3565 were isolated from glioblastoma and differentiated into differentiated glioma cell (DGC) 387 and DGC3565, respectively; GSC387 was divided into CatG group and CatG inhibitor group, and cells in the CatG group and CatG inhibitor group were cultured with 0.1 μg/μL recombinant human leukocyte antigen (HLA)-A*02:01 and HLA-B*15:01 combined with 4 ng/μL CatG or 10 mol/L CatG inhibitor for 10 min, respectively; the expressions of HLA-A*02:01 and HLA-B*15:01 were detected by Thomas bright blue staining, and the protein expressions of major histocompatibility complex (MHC)-I and MHC-DR were detected by Western blotting. (3) GSC387, GSC3565, DGC387, and DGC3565 were divided into 4 groups, including CatG group, CatG inhibitory group, blank antibody group 1 and blank antibody group 2, respectively; 4 ng/μL CatG, 10 μmol/L CatG inhibitor, blank antibody 1 and blank antibody 2 were added into the cells from the 4 groups for 24 h, and the expression of HLA-ABC was detected by flow cytometry. (4) GSC387, GSC3565, DGC387, and DGC3565 were divided into CatG group and CatG inhibitory group, respectively; luciferase assay was used to detect the influence of CatG in the killing effects of T cells and natural killer cells. Results:(1) The survival rate in patients from CTSG mRNA high expression group was significantly higher than that in patients from CTSG mRNA low expression group, and the survival rate in patients from β2M mRNA low expression group was statistically higher than that in patients from β2M mRNA high expression group ( P<0.05); a negative correlation between CTSG mRNA and β2M mRNA expressions was noted in glioma tissues ( r=-9.160, P=0.000). (2) Thomas bright blue staining showed that the expressions of HLA-A*02:01 and HLA-B*15:01 obviously increased in the CatG group as compared with those in the CatG inhibitor group; Western blotting showed that as compared with the CatG inhibitor group, the CatG group had increased MHC-I expression, and decreased expressions of α and β chains of MHC-DR. (3) Flow cytometry showed that the HLA-ABC expressions in GSC387 and GSC3565 of the CatG group were statistically higher than those in the CatG inhibitor group ( P<0.05). (4) Luciferase assay showed that, as compared with the CatG inhibitor group, the CatG group had statistically higher proportion of T cells killing GSCs ( P<0.05). Conclusion:CatG can improve the immunotherapy efficacy in GSCs, mainly by increasing the MHC-I expression on the cell surface.