Objective To Discuss the causes,prevention and treatment of delayed bile leakage following laparoscopic cholecystectomy (LC). Methods From November 1994 to December 2007,totally 11 000 cases of LC were performed in our hospital,12 of them developed delayed bile leakage after the procedure. The mechanism,characteristics,treatment,and prognosis of the disease were analyzed retrospectively. Results Among the twelve cases of delayed bile leakage,eight cases showed perforation of the bile duct caused by thermoelectric effect,two cases had leakage of the aberrant bile-duct,and two cases were injured by unknown reasons. B-ultrasonography-guided drainage was performed in four of the patients,laparoscopic or open repair combined with T-tube drainage were carried out in three,abdominal puncture combined with endoscopic nasobiliary drainage (ENBD) was done in three,abdominal puncture with endoscopic retrograde biliary drainage (ERBD) was made in one,and open repair and bilio-jejunal anastomosis was performed in one. All patients were cured and discharged in 8 to 21 days with a mean of 15 days. No intra-abdominal infection,hemorrhage,recurrent bile leakage or intestinal leakage occurred after the operations. Re-examination by B-ultrasonography or cholangiopancreatography performed ten days after the drainage showed no stricture or dilation of the bile duct,and contrast media drained out fluently. The 12 cases were followed up for one year,none of them showed biliary stenosis or infections during the period. Conclusions Post-LC injury of the bile duct is mostly caused by thermoelectric effect. Gentle operation and separation strictly following the anatomical levels are the keys in prevention of postoperative delayed bile leakage; while endoscopy and interventional treatment are main methods for the diagnosis and treatment of the disease.

Objective To construct the vector that expresses the fusion protein of HIV-Tat protein and red fluorescent protein(mCherry) in mammalian cells,and observe by fluorescence microscopy the intracellular transduction and localization of recombinant protein in cells,in order to obtain a useful tool for the study of the uptake mechanism and intracellular localization of HIV-TAT.Methods With the designed primer coding mCherry sequence,the mCherry gene was amplified by PCR with the vector pmCherry-C2 as template,and inserted into vector pET14b-His-TAT to construct the expression vector pET14b-His-TAT-mCherry.The constructed vector was then transformed into E.coli BL21(DE3),which had been identified by PCR and double digested with restriction endonuclease,followed by sequencing.After IPTG induction,the recombinant protein of His-TAT-mCherry was lyzed and analyzed with SDS-PAGE.Purified His-TAT-mCherry recombinant protein was added to Hela cells and the fluorescence was observed to evaluate the transduction efficiency.Results The results of identification by PCR,digestion with restriction endonuclease and sequencing indicated that the vector His-TAT-mCherry was correctly constructed.His-TAT-mCherry fusion protein was expressed in mammalian Hela cell line and purified successfully,and the fusion protein showed cellular transduction activity.It was found by fluorescence microscopy that the red fluorescence protein located mainly over the cytoplasm,and also the membrane to some extent.Conclusion The expression vector is successfully constructed for HIV-TAT labeled with mCherry sequence.Effective expression and purification of this fusion protein is achieved.It has been observed that the constructed vector may be expressed in mammalian Hela cell under active condition.Thus,it might be useful in the study of uptake mechanism and intracellular localization of HIV-TAT.

OBJECTIVE To investigate the distribution and drug resistanc of nosocomial infection pathogens from lower respiratory tract in senile patients in different season.METHODS The sputum samples from lower respiratory tract infection in senile patients in two years,were collected to identify pathogens and drug sensitivity test j udged according to NCCLS standard.RESULTS The gram-negative bacilli accounted for 85.7%.The gram- posutive bacilli accounted for 14.3%.The predominant pathogens were Pseudomonas aeruginosa(20.3%),Kleb- siella pneumoniae(14.5%),Acinetobacter lwof fi(9.8%),coagulase-negative staphylococci(9.0%).The distribution and antibiotic-resistanc of main pathogens had different characteristics in different season. CONCLUSIONS Understanding the characteristics of local pathogen spectruma and the antibiotic-resistanc of main pathogens in different season were signincant on prevention and therapy of the lower respiratory tract infection in senile patients.

OBJECTIVE To investigate the distribution and drug resistance of nosocomial infection pathogens from lower respiratory tract in senile patients. METHODS The sputum and lower respiratory tract secretion in the senile patients with lower respiratory tract infection were collected nearly five years,and identified.The drug sensitivity test,the results of examination were judged according to NCCLS standard. RESULTS The Gram-negative bacilli accounted for 79.5%.The Gram-positive bacteria accounted for 20.5%.The predominant pathogens were Pseudomonas aeruginosa(19.6%),coagulase-negative Staphylococci(CNS)(18.7%),and acinetobacter lwoffi(11.6%).The resistant bacteria were markedly increasing. CONCLUSIONS The Gram-negative bacilli are the major pathogens in the senile deceased patients.The incidence of CNS infection is markedly increasing.The isolating rate of meticillin-resistant CNS is 100.0%.

Objective To analyze the clinical characteristics,treatment and prognosis of primary esophageal small cell carcinoma (PESC).Methods Fifty-five patients with PESC were analyzed retrospectively.The survival rate was calculated by the Kaplan-Meier method and the Log-rank test using SPSS 17.0 software.Results The median survival time of 55 patients was 12 months.The 6,12,24.36months survival rates of these patients were 87.3 ％ (48/55),50.9 ％ (32/55),12.7 ％ (7/55),7.3 ％ (4/55),respectively.In multivariate analysis,stage and type of treatment were shown to be independent predictive.Conclusion PESC is a malignant tumor with early metastasis and poor prognosis.Combined therapy may improve the survival time of PESC patients.

Objective To evaluate the changes in sagittal profiles and complications during treatment with growing rods (GRs) in hyperkyphotic early-onset scoliosis (EOS).Methods From December 2009 to December 2016,a total of 32 EOS patients who received growing rods treatment in our center,including 8 males and 24 females,were reviewed retrospectively.All the patients had minimum 2-year follow-up and over 2 lengthenings.Based on the reference value of thoracic kyphosis (TK) in T2-12 of normal children,the patients were categorized into an N group (20°≤TK≤50°,15 cases,4 males and 11 females) or K group (TK≥50°,17 cases,4 males and 13 females).The distribution of etiology was similar between the two groups.The average age was (6.2±2.0) years and (6.3±2.3) years respectively,curve flexibility was 34.6％± 10.4％ and 35.8％± 11.2％ before surgery.The precontoured rods were tunneled submuscularly,connecting proximal and distal anchors,and tandem or domino connectors.The rods were then locked after applying direct distraction that allowed appropriate elongation.The connectors were all placed under the deep fascia.Results The mean follow-up in the N and K groups was (5.5±1.9) years and (5.5±2.1) years,respectively.The distribution of proximal and distal anchors was similar between the two groups.The N and K groups,respectively,had an average number of lengthenings of 5.1±2.0 and 5.3±2.3,with mean lengthening intervals of (11.3±2.3) months and (10.9±1.9) months,respectively.In the N group,TK was decreased from 36.0°±9.4° to 30.6°±.8.3° after surgery,and to 32.2°±7.8° at the last follow-up,demonstrating it was maintained within the normal range.In the K group,TK was markedly reduced from 67.6°±11.6° to 41.7°±8.7° after the index surgery,with a correction rate of 38.3％± 14.6％,and the difference was statistically significant.And then it slightly increased to 46.5°±8.4° at the last follow-up,with correction loss of 7.1％±4.2％,and the difference was not statistically significant compared with the postoperatiom.The complication rate in the K group was significantly higher than in the N group (76.5％ vs.33.3％,P=0.031).The most common implant-and alignment-related complication in both groups was rod fracture (15.6％) and proximal junctional kyphosis (21.9％),respectively.The incidence of rod fracture in the N group and K group was 6.7％ and 23.5％,respectively.And the incidence of proximal junctional kyphosis was noted as 13.3％ and 29.4％ in the N group and K group,respectively.Proximal junctional angle (PJA) in the K group was greater than that in the N group preoperatively,postoperatively and at the last follow-up.Moreover,the increasing amount of PJA was significantly greater in the K group compared to the N group (1.6°± 1.0° vs.0.7°±0.8°).Four and seven complication events in the N and K groups,respectively,were evaluated with Grade Ⅰ.Four and seven complication events in the N and K groups,respectively,were classified as Grade Ⅱ A.Conclusion GRs can effectively restore the sagittal profile in hyperkyphotie EOS patients,but with a higher complication rate compared to the patients with normal kyphosis.

Objective To investigate the value of quantitative parameters of CEUS in predicting the effect of ultrasound ablation of uterine fibroids.Methods Sixty-three patients with uterine fibroids underwent CEUS before operation.The software of gray level measurement in JC HIFU system was used to obtain quantitative parameters of uterine fibroids perfusion.According to postoperative enhanced MR examination,the results were divided into satisfactory group (ablation rate≥70 ％) and partial ablation group (ablation rate＜ 70 ％).CEUS parameters of two groups were compared,and the correlation between parameters and ablation rate were analyzed.The predictive value was evaluated by ROC curve.Results There were significant differences in the time of arrival (AT),time to peak (TTP),enhancement time (ET),enhancement intensity (EI) and enhancement rate (ER) between the two groups (all P＜0.01).The AT,TTP and ET were positively correlated with the ablation rate (all P＜0.01),and the EI and ER were negatively correlated with the ablation rate (both P＜0.001).The area under the curve (AUC) of AT,TTP,ET,EI,ER were all greater than 0.7,among which ER had the highest predictive value (AUC-0.935).Using ER=5.23 as the cut-off point,the sensitivity of predicting ablation effect was 80.0％,specificity was 95.8％.Conclusion There is a certain correlation between the ultrasound ablation effect of uterine fibroids and the quantitative parameters of CEUS.ER can be used as a good predictor of ablation effect.

Objective: To investigate the short term clinical efifcacy of endovascular repair for complicated acute type Stanford B aortic dissection.
Methods: To retrospectively analyze the clinical data of 36 patients with complicated acute type Stanford B aortic dissection who received endovascular repair in our hospital from 2010-01 to 2014-06 including operational procedure and post-operative follow-up of CT angiography. There were 27 male and 9 female patients with the average age of 43.7 years (41-62) years.
Results: Successful operations were conducted in all 36 patients. 22 patients received endovascular repair combined with covering left subclavian artery (LSA),10 received endovascular repair combined with chimney technique, 2 received endovascular repair combined with vascular prosthesis bypass from left common carotid artery to LSA, 2 received endovascular repair combined with vascular prosthesis bypass from right common carotid artery to left common carotid artery, whose proximal parts were ligated. Viscera artery and lower extremity artery supply were restored gradually. No complication of endoleak occurred. There 30/36 (83.33%) patients were followed-up for 1 year, and 10 patients developed thrombus in full false lumen and 20 developed thrombus in partial false lumen after 1 year. Compared with pre-operative values, thoracic aortic true lumen volume increased in either thrombus in full false lumen (190 ± 68.7) ml vs, (125.3 ± 63.4) ml and thrombus in partial false lumen (166.2 ± 71.8) ml vs (110.1 ± 62.7) ml,P<0.001; thoracic aortic false lumen volume decreased (65.0 ± 67.4) ml vs (185.3 ± 66.6) ml and (132.3 ± 62.6) ml vs (224.5 ± 72.3) ml,P<0.001. Compared with pre-operative values, for patients with thrombus in full false lumen, the abdominal aortic true lumen volume increased (55.5 ± 12.4) ml vs (48.6 ± 12.2) ml,P<0.01; for patients with thrombus in partial false lumen, the abdominal aortic false lumen volume also increased (58.2 ± 21.5) ml vs (42.5 ± 18.5) ml,P<0.01.
Conclusion: For endovascular repair of complicated aortic dissection, covering LSA with chimney technique and hybrid operation of small incision could extend anchor zone and expand the range of endovascular repair which may improve the effect and reduce the complication for good short term effect.

OBJECTIVETo observe the effect of transcutaneous acupoint electrical stimulation (TAES) combined dexmedetomidine on hemodynamic of intracranial aneurysmal subarachnoid hemorrhage patients undergoing intervention, and their protection for brain Injury.

METHODSTotally 108 intracranial aneurysmal subarachnoid hemorrhage patients undergoing intervention were randomly assigned to the electroacupuncture (EA) group and the control group according to random digit table, 54 in each group. All patients were anesthetized with dexmedetomidine. Patients in the EA group were needled at bilateral Neiguan (PC6), Lieque (LU7), and Yunmen (LU2). Parameter setting was as follows: The dilatational wave at 1. 5 Hz, strength 2 - 4 mA, 30 min. Systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), and heart rate (HR) were compared between the two groups immediately after entry into the room (T0), after administration (T1), intubating (T2), resuscitation (T3), extubation (T4), and leaving the operating room (T5). Levels of S100β protein (S100β) and neuron specific enolase (NSE) were compared between the two groups at T0, immediately after surgery (T6), 6 h after operation (T7), 12 h after operation (T8), and 24 h after operation (T9).

RESULTSCompared with the same group at T0, SBP, DBP, MAP, and HR were significantly reduced in the two groups at T1-T5(P <0. 05), serum levels of S100β and NSE in the two groups were significantly increased at T6-T9 (P<0. 05). Compared with the control group at T1 - T5, SBP, DBP, MAP, and HR decreased in the EA group (P <0. 05). Compared with the control group at T6-T9, serum levels of S100β and NSE decreased in the EA group (P <0. 05).

CONCLUSIONTAES combined dexmedetomidine could effectively maintain stable hemodynamics of intracranial aneurysmal subarachnoid hemorrhage patients undergoing intervention, and regulate their serum levels of S100β and NSE.

Acupuncture Points ; Airway Extubation ; Blood Pressure ; Brain Injuries ; therapy ; Electric Stimulation ; Electroacupuncture ; Heart Rate ; Hemodynamics ; Humans ; Phosphopyruvate Hydratase ; S100 Calcium Binding Protein beta Subunit ; Transcutaneous Electric Nerve Stimulation

The aim of this study is to prepare hyaluronic acid (HA) modified core-shell liponanoparticles (pHA-LCS-NPs) as gene delivery system and investigate its gene transfection efficiency in human retinal pigment epithelium (ARPE-19) cells in vitro. The pHA-LCS-NPs was prepared by firstly hydrating dry lipid film with CS-NPs suspension to get LCS-NPs, then modifying the lipid bilayer with HA by amidation reaction between HA and dioleoyl phosphatidylethanolamine (DOPE). Its morphology, particle size and zeta potential were investigated. XTT assay was used to evaluate the cell safety of different vectors in vitro. The gene transfection efficiency of pHA-LCS-NPs modified with different contents of HA was investigated in ARPE-19 cells with green fluorescent protein (pEGFP) as the reporter gene. The results showed that the obtained pHA-LCS-NPs exhibited a clear core-shell structure with the average particles size of (214.9 +/- 7.2) nm and zeta potential of (-35 +/- 3.7) mV. The 24 h cumulative release of gene from pHA-LCS-NPs was less than 30%. After 48 h incubation, gene transfection efficiency of pHA-LCS-NPs/pEGFP was 1.81 times and 3.75 times higher than that of CS-NPs/pEGFP and naked pEGFP, respectively. Also no obvious cytotoxicity was observed on pHA-LCS-NPs. It suggested that the pHA-LCS-NPs might be promising non-viral gene delivery systems with high efficiency and low cytotoxicity.