Objective To obtain sufficient recombinant multiepitope peptide of ESAT-6.Methods DNA encoding ESAT-6 peptide was amplified by PCR using a pair of primers which were designed in accordance with the reported ESAT-6 gene sequence.The ESAT-6P gene was inserted into the prokaryotic expression vector pET-30a with the N-terminal 6His-tag.The recombinant peptide vector pET30a-ESAT-6P was transformed into E.coli BL21(DE3) via chemical transformation.The positive clone was screened by means of PCR.The target peptide was expressed in E.coli after induction with IPTG and analyzed by SDS-PAGE.The immunogenicity of the peptide was evaluated by dot immunobinding assay.Results The target peptide was successfully expressed and purified.The solubility analysis showed that the recombinant peptide existed as inclusion body in the E.coli.DIBA indicated that the ESAT-6P was specifically reactive to sera from TB patients.Conclusions The recombinant multiepitope peptide of ESAT-6 engineering bacteria has been obtained,which will be quite helpful to develop novel specific diagnostic reagents and effective vaccine.

ObjectiveTo investigate the role of T-type calcium channel in the spinal neurotoxicity of intrathecal (IT) lidocaine in rats.MethodsForty-eight adult male SD rats in which IT catheter was successfully implanted,weighing 230-270 g,were randomly divided into 4 groups ( n =12 each):dimethyl sulfoxide (DMSO)group (group D),lidocaine group (group L),mibefradil + lidocaine group (group M),normal saline + lidocaine group (group N).Another 12 rats served as control group (group C).DMSO and 10％ lidocaine 20μl were injected intrathecally in groups D and L respectively.After mibefradil 200 μg/10μl and normal saline 10 μl were injected intrathecally in groups M and N respectively,10％ lidocaine 20 μl was injected intrathecally in the two groups.The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured before IT injection and at 2,4,8 and 12 h and 1,2,3,4 and 5 d after IT injection (T0-9).Four rats were sacrificed at T6 in each group and their lumbar enlargements were removed for microscopic examination.ResultsCompared with group C,no significant change in MWT and TWL was found at each time point in group D,MWT was significantly increased at T1-8 and TWL was significantly prolonged at T1-7 in groups L and N,and MWT was significantly increased at T1-6 and TWL was significantly prolonged at T1-6 in group M ( P ＜ 0.05 ).Compared with groups L and N,MWT was significantly decreased at T1-4 and TWL was significantly shortened at T1-4 in group M ( P ＜ 0.05).Pathological injury was significantly reduced in group M as compared with groups L and N.ConclusionT-type calcium channel is involved in the spinal neurotoxicity of IT lidocaine in rats.

AIM　To investigate the effect of hydroxyl camptothecinum(HCPT) on pharmocokinetics of cyclosprine A(CsA) and the membrane lipid fluidity (LFU)of red cell in rabbits. METHODS　The whole blood concentration of CsA in rabbits was monitored using method of FPIA. The LFU of red cell was measured using method of DPH fluorescence polarization technique. RESULTS　The pharmocokinetic parameters of CsA in group CsA+HCPT such as a,V(c),K12 and CL(s) were significantly lower and T1/2(a) was significantly higher (P<0.05) than in group CsA. The concentration of CsA of group CsA+HCPT was higher after injection 0.25, 0.5 and 1.0 h, but have significantly high at 1.0 h point (P<0.05). After injection 0.5,1.0, 1.5 h the LFU of red cell in group CsA+HCPT was higher than in group CsA (P<0.05). The LFU in group CsA was lower that in control at 0.5,1.0 and 1.5 h point (P<0.05). CONCLUSION　HCPT can enhance the whole blood concentration of CsA in rabbits. CsA administrated with HCPT is feasible.

Objective To observe the clinical effects of Dexmedetomidine in combination with Sevoflurane for hysteroscopic resection. Methods 72 patients underwent hysteroscopic resection from January 2014 to December 2015 were divided into study group and control group according to the anesthesia mode. Patients in study group were in combination anesthesia mode of intravenous infusion of Dexmedetomidine and Sevoflurane inhalation, while patients in control group received Sevoflurane inhalation only. The clinical parameters and adverse reactions were observed and studied. Results Compared with control group, significant differences were shown in patients heart rate at T2, T3 and T4 and breathing rate at T2 in observation group (P < 0.05). Compared with T1 phase, there was statistical significance in study group patients systolic blood pressure (SBP), diastolic blood pressure (DBP), RR at T2, HR at T3, T4 (P < 0.05); Compared with control group, patients in study group had short onset time of anesthesia and long recovery of consciousness, while there has no statistical significance of the two groups (P > 0.05). Compared with control group, the respiratory depression, sedative effect of Ramsay sedation score, wakefulness, alertness / sedation score of observation method (OAA/S), digital pain score (NPRS), postoperative uterine contraction pain score, postoperative nausea and vomiting of study group had significant statistical differences (P < 0.05). Conclusion Combination anesthesia mode of intravenous pumping Dexmedetomidine and inhalation of Sevoflurane has good clinical effects for patients undergoing hysteroscopic resection.

AIM To investigate the effect of hydroxyl camptothecinum(HCPT) on pharmocokinetics of cyclosprine A(CsA) and the membrane lipid fluidity (LFU)of red cell in rabbits. METHODS The whole blood concentration of CsA in rabbits was monitored using method of FPIA. The LFU of red cell was measured using method of DPH fluorescence polarization technique. RESULTS The pharmocokinetic parameters of CsA in group CsA+HCPT such as a, V (c), K 12 and CL (s) were significantly lower and T 1/2(a) was significantly higher ( P

Objective To discuss the repairing methods of the wound after upper lip lesion excision.Methods The wound after upper lip lesion excision was repaired by expanded pedicled submental flap.The 3 cm-long incision was located in 1 cm to sub-mandible.The 100 ml expander was placed beneath the platysma,and the aqueducts and spigots of the expanders were laid out of the skin.After complete expansion,the spastic scars of the upper lip and nasal bottom were resolved,the nasal columella and upper lip were put back to the normal position.The pedicled submental flap was transferred to the wound after upperlip excision according to the size of the wound.The pedicle was severed after 3 weeks.Results There were 5 cases of the expanded pedicled submental flap to repair the wound after upper lip excision.The flap survived without complications.The appearances were satisfied by the patients.Conclusions The method of the expanded submental flap is suitable for the wound after upper lip excision.

Objective To evaluate the effects of dexmedetomidine on the cellular immune function of the rats with scald.Methods Seventy-two healthy male Sprague-Dawley rats,weighing 200-220 g,aged 120-150 days,were randomly divided into 3 groups (n =24 each):normal control group (group C),scald group (group S) and dexmedetomidine group (group D).Thirty percent of the total body surface was shaved and then exposed to 94 ℃ water for 12 s in S and D groups.The rats were resuscitated according to Parkland formula after scald in S and D groups,and in addition,dexmedetomidine 30 μg/kg was also intraperitoneally injected immediately after scald in D group.Before the model was established (T1) and at 12 and 24 h after scald (T2,3),blood samples from the inferior vena cava were collected for determination of T lymphocyte subsets CD3 +,CD4 + and CD8 +,NK cell,C-reactive protein (CRP),interleukin-6 (IL-6),IL-10 and tumor necrosis factor-alpha (TNF-α) level.CD4+/CD8+ was calculated.Arterial blood samples were collected for blood gas analysis.Results Compared with C,the CD3+,CD4+ and NK cell levels,CD4+/CD8+,pH value,PaCO2 and PaO2 were significantly decreased,and CD8+ levels,IL-6,IL-10,TNF-α,CRP and BE negative value were increased at T2,3 in S and D groups.Compared with group S,the CD3+,CD4+,NK cell and IL-10 levels,CD4+/CD8+,pH value,PaCO2 and PaO2 were significantly increased,and CD8+ levels,IL-6,TNF-α,CRP and BE negative value were decreased at T2,3 in group D.Conclusion Dexmedetomidine can improve the cellular immune function of the rats with scald.

Objective To investigate the application of the frontal branch of superficial temporal vessels pedicled flap in repairing lower eyelid ectropion.Methods Eight cases were collected from patients diagnosed with lower eyelid ectropion in our hospital from April 2012 to April 2015.In phase 1 of operation,the dilators were implanted into the frontal branch of superficial temporal vessels and fully expanded by normal saline injection;In phase 2,the scar of lower eyelid was incised,and the expanded forehead flaps were transferred to cover the wound after the lower eyelid released back to normal anatomy location;In phase 3,the flap delay operation was manipulated 3 weeks after phase 2,and the left wound after scar excision was finished by pedicle division 1 week later.Results All patients in the study showed a good appearance and function of lower eyelid.There were no complications such as flap congestion and necrosis occurred.Meantime there were no relapses observed according to the follow-ups ranging from 6 months to 1 year.Conclusions The application of the frontal branch of superficial temporal vessels pedicled flap shows a promising procedure in treatment of lower eyelid ectropion.

ObjectiveTo explore the clinical application of laparoscopic choledochoduodenstomy in Hepatobiliary Surgery Department.MethodsTwenty-six patients of laparoscopic choledochoduodenstomy were retrospectively analyzed between January 2007 and September 2011 in Hepatobiliary Surgery Department of PLA General Hospital,including 8 cases of choledochalcyst,11 cases of bile duct stone and 7 cases of bile duct cancer.ResultsAll of patients were successfully performed operation of total laparoscopic choledochal cyst excision.The operation time was 60 to 110 minutes.And the time of hospital stay was 3 to 7 days.One patient was found anastomotic stoma stricture,the other cases had no post-operative complication.There was no patient dead.ConclusionLaparoscopic choledochoduodenstomy needs mastering the indication of operation and laparoscopic anastomosis technique.Under this condition,the operation will be safe and feasible.

Objective To evaluate the role of C-Jun N-terminal kinase (JNK) signal transduction pathway in spinal neurotoxicity induced by lidocaine in rats.Methods Seventy-two adult male Sprague-Dawley rats,weighing 220-260 g,were randomly divided into 6 groups (n =12 each):control group (group Ⅰ),sham operation group (group Ⅱ),JNK inhibitor group (group Ⅲ),dimethyl sulfoxide (DMSO) group (group Ⅳ),lidocaine group (group Ⅴ),and JNK inhibitor and lidocaine group (group Ⅵ).Group Ⅰ received no treatment.Intrathecal catheter was placed in the subarachnoid space in group Ⅱ.SP600125 25 μg and DMSO 20 μl were injected intrathecally in Ⅲ and Ⅳ groups,respectively.In group Ⅴ,10％ lidocaine 20 μl was intrathecally injected.SP600125 25 μg was injected intrathecally and 30 min later 10％ lidocaine 20 μl was injected intrathecally in group Ⅵ.Paw withdrawal threshold to yon Frey filament stimulation (PWT) and paw withdrawal latency to nociceptive thermal stimulation (PWL) were measured before intrathecal catheter was implanted (T0),before intrathecal administration (T1) and at 4,8 and 12 h and on 1,2,3,4,5 and 6 days after intrathecal administration (T2-10).At 24 h after intrathecal administration,4 rats were randomly chosen from each group and sacrificed.Their lumbar enlargements were removed for determination of phosphorylated JNK (p-JNK) expression (using Western blot) and neuronal apoptosis (by TUNEL).The apoptotic index was calculated.Results Compared with group Ⅰ,no significant difference was found in MWT and TWL in Ⅱ,Ⅲ groups and expression of p-JNK in Ⅱ and Ⅳ groups (P ＞ 0.05),MWT at T2-4,6-8 and TWL at T2-4,7 in group Ⅴ and MWT at T2-6 and TWL at T2-5 in group Ⅵ were significantly increased,the expression of p-JNK was down-regulated and the apoptotic index was decreased in group Ⅲ (P ＜ 0.05),and the expression of p-JNK was up-regulated and the apoptotic index was increased in Ⅴ and Ⅵ groups (P ＜ 0.05).Compared with group Ⅴ,MWT and TWL were significantly decreased,the expression of pJNK was down-regulated and the apoptotic index was decreased in group Ⅵ (P ＜ 0.05).Conclusion Activation of JNK signal transduction pathway is involved in spinal neurotoxicity induced by lidocaine in rats possibly through promoting neuronal apoptosis in the spinal cord.