Objective To observe the effect of patient-controlled intravenous analgesia (PCIA)with dezocine combined with sufentanil on inflammatory response and pain after laparoscopic hepatectomy for hepatocellular carcinoma.Methods Sixty patients (43 males,17 females,aged 18-60 years,ASA grade Ⅰ or Ⅱ) scheduled for laparoscopic hepatectomy for hepatocellular carcinoma were divided into sufentanil group (group S) and dezocine+sufentanil group (group DS) according to the random number table,n=30 each.Patients in group S were given 100 ml normal saline containing sufentanil 2.0 μg/kg and tropisetron 5 mg.Patients in group DS were given 100 ml normal saline containing sufentanil 2.0 μg/kg,dezocine 0.5 mg/kg and tropisetron 5 mg.VAS scores and numeric sedation scale (NSS) scores were recorded at 4,24,48 h after operation and patients' satisfaction scores were recorded at 48 h after operation.The levels of serum tumor necrosis factor-α (TNF-α),interleukin-2 (IL-2),interleukin-6 (IL-6) in blood samples harvested before induction of anesthesia and 0,4,24 and 48 h after operation were measured by ELISA.The times of efficient injection and incidence of adverse effect within 48 h after operation were recored.Results Compared with group S,the VAS scores in group DS were decreased significantly while the satisfaction of patients to analgesia were increased significantly at 4,24,48 h after operation (P<0.05).There were no obvious differences in NSS scores between two groups.Compared with before induction of anesthesia,the concentrations of TNF-α and IL-6 were increased significantly while the concentrations of IL-2 was decreased significantly in both groups at 4,24,48 h after operation (P<0.05).Compared with group S,the concentrations of TNF-α and IL-6 were decreased significantly while the concentrations of IL-2 was increased significantly in group DS at 24,48 h after operation (P<0.05).The times of efficient injection in group DS were less than that in group S significantly within 48 h after operation [(2.0±0.7) times vs.(7.2±1.3) times] (P<0.05).There were no obvious differences in adverse effects between two groups.Conclusion PCIA with dezocine 0.5 mg/kg combined with sufentanil 2.0 μg/kg can alleviate the inflammatory response to some extent in patients after laparoscopic hepatectomy for hepatocellular carcinoma,and it can offer a safe and effective analgesic effect.

Objective To construct a eukaryotic expression plasmid vector encoding Cbl-b gene-specific short hairpin RNAs (shRNAs),and to evaluate its interference effect,so as to lay a foundation for further study on the role of Cbl-b in the immunotherapy of malignant melanoma.Methods According to the sequence of Cbl-b cDNA,4 pairs of shRNAs targeting the Cbl-b gene were designed and synthesized,and then inserted into the plasmid PGPU6/GFP/Neo to construct recombinant plasmids.After identification by DNA sequencing,the 4 shRNA expression vectors were cotransfected into 293T cells with the Cbl-b gene eukarytic expresson plasmid,respectively.The knockdown efficiency of these shRNA expression plasmids on Cbl-b expression was evaluated by real-time (RT) fluorescence-based quantitative PCR and Western blot at 48 hours aftert transfection.Results Sequencing analysis revealed that all the 4 pairs of shRNAs were successfully inserted into the eukarytic expression vector PGPU6/GFP/Neo.As RT-PCR and Western blot showed,all the 4 shRNA-expressing vectors could downregulate Cbl-b expession,and the NO.1 shRNA-expressing vector displayed the strongest interference effect(P ＜ 0.05).Conclusions A eukaryotic expression plasmid vector was successfully constructed for Cbl-b gene-specific shRNAs,and the most effective shRNA was selected in this study.

Objective To evaluate the effect ofT-cell immunoglobulin and mucin domain-3 (TIM-3) on TRP-2180-188 peptide-stimulated murine spleen lymphocytes co-cultured with B16F10 murine melanoma cells.Methods A recombinant plasmid pFUSE-TIM-3-mIgG2Aae1-Fc2 encoding TIM-3 was constructed.Then,the recombinant plasmid and an empty plasmid pFUSE-mIgG2Aae1-Fc2 were transfected into human 293T epithelial cells followed by 48-hour culture for the preparation of supernatants containing TIM-3 and Ig-tail respectively.C57BL/6 mice were immunized with the TRP-2180-188 peptide vaccine for 4 sessions.One week after the last vaccination,C57BL/6 mice were sacrificed,and spleen lymphocytes were collected and then cultured with the TRP-21180-188 peptide and interleukin-2 (IL-2) for 5 days,with lymphocytes untreated with the TRP-2180-188 peptide or IL-2 serving as the control group.Mitomycin-treated B16F10 murine melanoma cells and TRP-2180-188 peptide-stimulated lymphocytes were co-cultured with the presence of supernatants of 293T cells that had been cultured for 48 hours (blank control group),TIM-3-containing supernatants (TIM-3 group) and Ig-tail-containing supernatants (negative control group) separately.After 24 and 48 hours of co-culture,cell counting kit-8 (CCK-8) assay was performed to estimate the proliferative activity of lymphocytes,enzyme-linked immunosorbent assay (ELISA) to determine the supernatant levels of interferon (INF)-γ and tumor necrosis factor (TNF)-α,flow cytometry to determine the percentage of CD8 + T cells in the co-culture system.Results Enzyme digestion and sequence analysis showed that the TIM-3 gene was successfully inserted into the eukaryotic expression plasmid.After 48-hour culture,TIM-3 and Ig-tail expressions were detected in the supernatants of 293T cells transfected with the recombinant plasmid and empty plasmid respectively.As CCK-8 assay showed,the proliferative activity of lymphocytes was significantly lower in the TIM-3 group than in the blank control group and negative control group after 24-and 48-hour culture (78.06％ ± 6.37％ vs.100.00％ ± 10.42％ and 108.70％ ± 9.90％ at 24 hours,42.93％ ± 5.93％ vs.100.00％ ± 6.24％ and 168.00％ ± 2.98％at 48 hours,all P ＜ 0.05),so was the ratio of cellular proliferative activity at 48 hours to that at 24 hours (all P ＜ 0.05).Compared with the blank control group and negative control group,the TIM-3 group showed significantly decreased supernatant levels of IFN-γ and TNF-α after 24-hour (IFN-γ:192.96 γ 5.05 ng/L vs.216.44 ± 7.85 ng/L and 223.67 ±7.79 ng/L,both P＜ 0.05;TNF-α:58.43 ± 0.26 ng/L vs.26.43 ± 0.01 ng/L and 86.85 ± 1.12 ng/L,both P＜ 0.05) and 48-hour culture (IFN-γ:54.95 ± 0.57 ng/L vs.230.06 ± 4.23 ng/L and 167.24 ± 3.33 ng/L,both P ＜ 0.05;TNF-α:30.23 ±0.26 ng/L vs.26.84 ± 0.20 ng/L and 45.34 ± 0.22 ng/L,both P ＜ 0.05).In addition,the median percentage of CD8+ T cells was significantly increased in the TIM-3 group compared with the blank control group and negative control group after 24-and 48-hour culture (3.30％ vs.0.421％ and 2.22％ at 24 hours,4.06％ vs.0.577％ and 0.691％ at 48 hours,all P＜ 0.05).Conclusion TIM-3 in vitro can suppress the proliferative activity of and secretion of IFN-γand TNF-α by lymphocytes,but increase the percentage of CD8 + T cells in the co-culture system of TRP-2180-188 peptide-stimulated lymphocytes and B16F10 cells.

Objective To compare the kidney injury in diabetic patients undergoing liver cancer resection performed under different methods of anesthesia.Methods Sixty diabetic patients of both sexes,aged 40-64 yr,weighing 48-75 kg,of ASA physical shatus Ⅱ or Ⅲ (liver function Child-Pugh grade A),scheduled for elective liver cancer resection,were randomly divided into 2 groups (n =30 each) using a random number table:total intravenous anesthesia with propofol group (group P) and combined intravenous-inhalational anesthesia with sevoflurane group (group S).In group S,8％ sevoflurane was inhaled (FGF 8 L/min),and sufentanil 0.4 μg/kg and cisatracurium besylate 0.2 mg/kg were injected intravenously after the patients lost consciousness.In group P,propofol 1-2 mg/kg,sufentanil 0.4 μg/kg and cisatracurium besylate 0.2 mg/kg were injected intravenously.The patients were tracheally intubated and mechanically ventilated.Anesthesia was maintained with inhalation of 2 ％-3 ％ sevoflurane (FGF 2 L/min) in group S,or with iv infusion of propofol 0.5-0.8 mg· kg-1 · h-1 in group P,and with iv sufentanil 10 μg and cisatracurium 0.1 mg/kg when needed in both groups.BIS value was maintained at 40-60 and PET CO2 at 35-45 mmHg during operation.Before induction of anesthesia,at the end of operation,and at 24 and 72 h after operation,blood samples were collected from the central vein for determination of the levels of serum creatinine,blood urea nitrogen,serum cystatin C and 24 h.urinary microalbuminuria.Results Compared with group S,the levels of serum cystatin C at 24 and 72 h after operation and 24 h urinary microalbuminuria were significantly increased,and no significant changes were found in the levels of serum creatinine and blood urea nitrogen at each time point in group P.Conclusion The kidney injury is reduced in the diabetic patients undergoing liver cancer resection performed under combined intravenous-inhalational anesthesia with sevoflurane as compared with that under total intravenous anesthesia with propofol.

Objective To evaluate the effects of sulforaphane preconditioning on cognitive dysfunction induced by sevoflurane anesthesia in aged rats.Methods Thirty Sprague-Dawley rats,aged 22 months,weighing 380-560 g,were randomly divided into 3 groups (n =10 each) using a random number table:control group,sevoflurane group (Sev group),and sulforaphane group (Sul group).In group Sul,sulforaphane25 mg/kg was administered by oral gavage once a day for 7 consecutive days,while the equal volume of distilled water was given instead of sulforaphane in Sev and C groups.Groups Sev and Sul inhaled 3％ sevoflurane in oxygen (2.5 L/min) and group C inhaled air (100 min/d) for 5 consecutive days starting from the end of gavage.At 24 h after sevoflurane inhalation,cognitive function was detected using Morris water maze and open field tests.The escape latency,frequency of crossing the original platform,the number of crossing the grid,the number of standing on the back legs and the time the animals spent in the central square were recorded.Results Compared with group C,the frequency of crossing the original platform,the number of crossing the grid,and the number of standing on the back legs were significantly reduced,the time the animals spent in the central square and escape latency on 1st day were prolonged and no significant changes were found in the escape latency on 2nd-4th days in group Sev.Compared with group Sev,the frequency of crossing the original platform,the number of crossing the grid,and the number of standing on the back legs were significantly increased,and the time the animals spent in the central square and escape latency on 1st day were shortened.Conclusion Sulforaphane preconditioning can improve the cognitive dysfunction induced by sevoflurane anesthesia in aged rats.

Objective To investigate the effect of sevoflurane anaesthesia on the cognitive function and phosphorylation of tau protein in hippocampal neurons in amyloid precursor protein (APP) transgenic mice.Methods Male APP gene mutation mice,weighing 18-22 g and aged 8-12 weeks,were used in this study.Forty-four APP positive mice were randomly divided into two groups:sevoflurane group (group AS,n =28) and control group (group AC,n =16).And other forty-four APP negative mice were randomly divided into two groups:sevoflurane group (group S,n =28) and control group (group C,n =16).The animals in groups S and AS inhaled 3％ sevoflurane for 4 hours.While in groups C and AC,the animals inhaled pure oxygen for 4 hours.Morris water maze was performed 24 hours after sevoflurane or pure oxygen inhalation.The phosphorylation of tau protein at Ser262 and Ser396 sites was detected by Western blotting on 1 day after pure oxygen inhalation in groups AC and C,and on 1,3 and 7 days after sevoflurane inhalation in groups AS and S.Results Compared with group C,the escape latency was significantly prolonged and the duration of staying at the original platform quadrant was shortened in groups S and AC,and the phosphorylation of tau protein at Ser262 site in group S and phosphorylation of tau protein at Ser262 and Ser396 sites in group AS were increased (P ＜ 0.05).Compared with group S,the escape latency was significantly prolonged,the duration of staying at the original platform quadrant was shortened,and the phosphorylation of tau protein at Ser262 and Ser396 sites was increased in group AS (P ＜ 0.05).Compared with group AC,the escape latency was significantly prolonged,the duration of staying at the original platform quadrant was shortened,and the phosphorylation of tau protein at Ser262 and Ser396 sites was increased in group AS (P＜0.05).Conclusion Sevoflurane anesthesia can aggravate the impairment of cognitive function in APP positive mice and the increase in the phosphorylation of tau protein at Ser262 and Ser396 sites is involved in the mechanism.

Objective To evaluate the effect of ketamine on the expression of tryptophan hydroxylase 2 (TPH2) in the median raphe nuclei of mentally depressed mice.Methods Thirty-six healthy SPF male C57BL/6J mice,aged 8-12 weeks,weighing 20-26 g,were divided into 3 groups (n=12 each) using a random number table:control group (C group),depression group (D group) and depression plus ketamine group (D+K group).Mental depression was induced by forcing the animals to swim in a narrow cylinder from which they can not escape.Ketamine 15 mg/kg was intraperitoneally injected once a day for 7 consecutive days starting from 1 day after successful establishment of the model in group D+K.The equal volume of normal saline was given instead of ketamine in C and D groups_ Forced swimming test was performed again at 30 min after the last administration,and the immobility time was recorded.Open field test was also performed at 30 min after the last administration,and the total horizontal distance and the number of standing on the back legs were recorded.The mice were sacrificed after the end of the behavioral testing,and the hippocampi and median raphe nuclei were isolated.High-performance liquid chromatography-electrochemical detection assay was used to measure the content of 5-hydroxytryptamine (5-HT) in hippocampi.The expression of TPH2 protein and mRNA in the median raphe nuclei was detected using Western blot and real-time polymerase chain reaction,respectively.Results Compared with group C,the immobility time was significantly prolonged,the total horizontal distance was shortened,the number of standing on the back legs and content of 5-HT in hippocampi were deceased,and the expression of TPH2 protein and mRNA in the median raphe nuclei was down-regulated in group D,and the total horizontal distance was significantly shortened,the number of standing on the back legs was decreased (P＜0.05),and no significant change was found in the immobility time,content of 5-HT in hippocampi or expression of TPH2 protein and mRNA in the median raphe nuclei in group D +K (P＞0.05).Compared with group D,the immobility time was significantly shortened,the content of 5-HT in hippocampus was increased,the expression of TPH2 protein and mRNA in the median raphe nuclei was up-regulated (P＜0.05),and no significant change was found in the total horizontal distance or the number of standing on the back legs in group D+K (P＞0.05).Conclusion The mechanism by which ketamine produces anti-depressant effect may be related to up-regulation of TPH2 expression in the median raphe nuclei and increase in the synthesis of 5-HT in hippocampi of mice.

Objective:To investigate the feasibility of periodontal mechanical therapy for chronic periodontitis and coronary heart disease patients with low dose of aspirin.Methods:Sixty nine chronic periodontitis patients with coronary heart disease were randomly selected as the experimental group (medication group,group A),the control group (withdrawal group,group B) including 20 chronic periodontitis patients with coronary artery disease,stopping the drug for one week and another control group with 50 chronic periodontitis patients (group C).The three groups were examined with pocket probing,and received supragingival scaling,subgingival scaling,and root planning.Local bleeding after operation was observed.In 30 minutes after periodontal mechanical treatment,there was still a need to take some hemostatic measures (containing the oxidized cellulose putting in the periodontal pocket,gauze oppressing,and suturing).Nd:YAG laser was used to stop bleeding 60 minutes after operation.Results:At baseline,there was no significant difference in the three groups,as to the plaque index (PLI),the probing depth (PD),and the attachment loss (AL).The bleeding index (BI)in group A was significantly higher than that in group C (P =0.024),higher than that in group B (P =0.088).The platelet maximum aggregation rate (Aggmax) was detected in some subjects.The average Aggmax value group A was 15.2％,which was much greater than that in group B (60.7％) and group C (62.5 ％).The three groups were all safe in the treatment of periodontal therapy.There were five cases of active bleeding in group A,one case in group B and one case in group C in 30 minutes after operation.In 60 minutes after operation,there was one case of bleeding actively in group A.Nd:YAG laser was used to stop bleeding successfully.Conclusion:The chronic periodontitis and coronary heart disease patients with long-term oral administration of low dose of aspirin can be safely treated with periodontal mechanical treatment,and the effect of local hemostasis is positive without stopping the drug.

Objective To evaluate the efficacy of high frequency two-lung ventilation (TLV) with low tidal volume assisted by CO2 pneumothorax for airway management in patients undergoing thoracoscopic radical resection of esophagus cancer.Methods Thirty patients of both sexes,aged 48-64 yr,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective thoracoscopic radical resection of esophagus cancer,were divided into 2 groups (n =15 each) using a random number table:onelung ventilation group (group O) and TLV group (group T).A left-sided double-lumen tube was inserted orally in group O,and a single-lumen tube was placed orally in group T.During thoracoscopic surgery,the left lung was ventilated,with tidal volume 8 ml/kg and respiratory rate 14 breaths/min in group O.In group T,artificial pneumothorax was induced by continuous CO2 insufflation with CO2 pressure at 10 mmHg,and bilateral lungs were ventilated,with tidal volume 5 ml/kg and respiratory rate 20 breaths/min.Mean arterial pressure and heart rate were recorded before induction of anesthesia,immediately after intubation (T1),at 10 min after intubation (T2),at 30 min after the start of thoracoscopic surgery (T3),immediately after the end of thoracoscopic surgery (T4) and at 30 min of TLV (T5).Arterial blood samples were collected for blood gas analysis at T2,T3,T4 and T5.The exposure of the surgical field and the number of lymph node dissection in the left recurrent laryngeal nerve chain were recorded during surgery.The emergence time,extubation time and time for recovery of consciousness were recorded.Results Arterial oxygen partial pressure was significantly lower at T3,4 than at T2 in the two groups,and arterial carbon dioxide partial pressure was significantly higher,and the pH value was lower at T3,4 than at T2 in group T (P＜0.05).Compared with group O,arterial carbon dioxide partial pressure was significantly increased,the pH value was decreased,and the number of lymph node dissection in the left recurrent laryngeal nerve chain was increased at T3,4 in group T (P＜0.05).There were no significant differences between the two groups in the good exposure of the surgical field,emergence time,extubation time and time for recovery of consciousness (P＞0.05).Conclusion High frequency TLV with low tidal volume when assisted by CO2 pneumothorax can serve as a feasible mode for airway management in patients undergoing thoracoscopic radical resection of esophagus cancer.

Objective To evaluate in vitro effects of specific small interfering RNA (siRNA)-silencing of the casitas B-lineage lymphoma b (Cbl-b)gene on immunocompetence of primary murine lymphocytes. Methods Spleens were resected from C57BL/6 mice, and splenic lymphocytes were sterily isolated and cultured in vitro. These lymphocytes were divided into 3 groups: silence group transfected with a Cbl-b-specific siRNA using the EntransterTM-R 4000 reagent, negative control group transfected with a negative control siRNA using the EntransterTM-R4000 reagent, blank control group receiving no treatment. After additional culture for 72 hours, ELISA was performed to measure levels of interferon γ(IFN-γ)and tumor necrosis factor α (TNF-α)in culture supernatants of lymphocytes. In addition, the Cbl-b gene-silenced lymphocytes were co-cultured with B16F10 melanoma cells to evaluate their immunocytotoxic effects on melanoma cells. Results Splenic lymphocytes were successfully isolated from C57BL/6 mice and cultured in vitro, and the Cbl-b-specific siRNA was also successfully transfected into the primary murine lymphocytes and effectively down-regulated the expression of Cbl-b gene in them. Compared with the negative control group and blank control group, the silence group showed significantly increased supernatant levels of IFN-γ and TNF-α(all P < 0.05). The immunocytotoxic effect of lymphocytes on melanoma cells was significantly stronger in the silence group than in the negative control group. Conclusion Cbl-b gene silencing can promote secretion of IFN-γ and TNF-α by murine lymphocytes, and enhance their immunocytotoxic effects on B16F10 melanoma cells in vitro.