Objective To investigate the effects of cold preservation on the expression of GATA in intrahepatic bile duct.Methods The intrahepatic bile duct tissues of SD rats were obtained by collagenase perfusion combined with mechanical separation.After being cut into fragments,the intrahepatic bile duct tissues were cultured in rat tail collagen gel for 48 hours before experiment.All the rats were divided into the control group,cold preservation 1 hour (CP1 h) group and cold preservation 12 hours (CP12 h) group.There were 5 rats in each group.The mRNA and protein expressions of GATA were detected by Real-Time polymerase chain reaction and Western blot.Measurement data with normal distribution were presented as (x) ± s.Comparison among 3 groups was done by ANOVA and pairwise comparisons were done by LSD test.Results The mRNA expressions of GATA3,GATA4,GATA6 were detected,while the mRNA expressions of GATA1,GATA2 and GATA5 were undetectable in intrahepatic bile duct tissue of the control group.The mRNA expressions of GATA4 in the CP1 h group,CP12 h group and the control group were 0.72 ± 0.08,0.56 ± 0.07 and 0.96 ± 0.06,with significant difference among the 3 groups (F =38.981,P ＜0.05).The mRNA expression of GATA4 in the CP12 h group was significantly lower than that in the CP1 h group and the control group,and the mRNA expression of GATA4 in the CP1 h group was significantly lower than that in the control group (P ＜ 0.05).The mRNA expression of GATA6 in the CP1 h group,CP12 h group and the control group were 0.83 ± 0.07,0.68 ± 0.12 and 0.98 ± 0.12,with significant difference among the 3 groups (F =10.175,P ＜ 0.05).The mRNA expression of GATA6 in the CP12 group was significantly lower than that in the CP1 h group and the control group,and the mRNA expression of GATA6 in the CP1 h group was significantly lower than that in the control group (P ＜ 0.05).The mRNA expressions of GATA3 in the CP1 h group,CP12 h group and the control group were 0.92 ± 0.06,0.89 ± 0.05 and 0.98 ± 0.11,with no significant difference among the 3 groups (F =1.674,P ＞ 0.05).The protein expressions of GATA4 in the CP1 h group,CP12 h group and the control group were 0.78 ± 0.07,0.64 ± 0.06 and 0.99 ± 0.10,with significant difference among the 3 groups (F =24.211,P ＜ 0.05).The protein expression of GATA4 in the CP12 h group was significantly lower than that in the CP1 h group and the control group,and the protein expression of GATA4 in the CP1 h group was significantly lower than that in the control group (P ＜ 0.05).The protein expressions of GATA6 in the CP1 h group,CP12 h group and the control group were 0.90 ± 0.04,0.75 ±0.06 and 0.98 ±0.11,with significant difference among the 3 groups (F=11.651,P＜0.05).The protein expression of GATA6 in the CP12 h group was significantly lower than that in the CP1 h group and the control group (P ＜ 0.05).Conclusion The expressions of GATA4 and GATA6 in the intrahepatic bile duct tissues are decreased significantly after cold preservation,which indicate that GATA4 and GATA6 might be involved in the pathophysiological process of the bile duct after cold preservation.

Objective To evaluate the intraoperative opioid-sparing effect of different frequency transcutaneous electrical acupoint stimulation (TEAS) in the patients undergoing video-assisted thoracoscopic pneumonectomy.Methods Eighty patients,aged 40-64 yr,weighing 50-90 kg,of ASA physical status Ⅰ-Ⅲ,scheduled for elective thoracoscopic pneumonectomy under general anesthesia,were randomly divided into 4 groups (n =20 each) using a random number table:control group (group Con),stimulation on Lieque (LU7)-Quchi (LI11)-Neiguan (PC6)-Hegu (LI4) at 2/100 Hz group (group 2/100 Hz),stimulation on LU7-LI11-PC6-LI4 at 2 Hz group (group 2 Hz),and stimulation on LU7-LI1 1-PC6-LI4 at 100 Hz group (group 100 Hz).The patients in group Con had the electrodes applied,but received no stimulation.In 2/100 Hz,2 Hz and 100 Hz groups,the patients received 2/100,2 and 100 Hz TEAS on LU7-LI11-PC6-LI4 acupoints ipsilateral to the surgery site,respectively,starting from 30 min before induction of anesthesia until the end of surgery,and the intensity was the maximum current that could be tolerated.Anesthesia was induced with iv midazolam,propofol,sufentanil and cisatracurim,and maintained with target-controlled infusion of remifentanil and propofol,continuous infusion of cisatracurim,and iv boluses of sufentanil when necessary.The target plasma concentration of propofol was adjusted to maintain BIS value at 40-60 during operation.The initial target effect-site concentration of remifentanil was 1 ng/ml,and adjusted to 4 ng/ml at skin incision.The concentration of remifentanil and consumption of sufentanil were adjusted to maintain Analgesia Nociception Index (ANI) at 50-70.When the concentration of remifentanil was increased to 4 ng/ml,ANI was still less than 50,and then 0.1 μg/kg sufentanil was given.The duration of operation and intraoperative consumption of remifentanil and sufentanil (the consumption of sufentanil was converted to the consumption of remifentanil producing the equivalent effect by 1:10) were recorded.Results The intraoperative consumption of remifentanil was significantly reduced in 2/100 Hz group as compared with Con,2 Hz and 100 Hz groups.There was no significant difference in the intraoperative consumption of remifentanil between Con group,2 Hz group and 100 Hz group.Conclusion The use of 2/100 Hz but not 2 and 100 Hz TEAS on LU7-LI11-PC6-LI4 significantly reduces intraoperative opioid consumption in the patients undergoing video-assisted thoracoscopic pneumonectomy.

The paper introduced the hospital's attempt to build a scientific and practical set of performance evaluation system based on KPI.The system determines the quantitative indicators for the efficiency and performance of each department,and effectively helps accomplish common objectives of the hospital and its departments,playing a key role in the sustaimble development of the hospital.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; immunology ; pathology ; Cytokines ; blood ; Esophageal Neoplasms ; immunology ; pathology ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-12 ; blood ; Interleukin-4 ; blood ; Interleukin-5 ; blood ; Lymphatic Metastasis ; Male ; Middle Aged ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Tumor Necrosis Factor-alpha ; blood

To construct an immortalized rat astrocyte strain genetically modified by rat preprogalanin gene (IAST/GAL) and detect its galanin (GAL) expression and secretion, a cDNA fragment of rat GAL in plasmid of pBS KS(+)-GAL was inserted into eukaryotic expression vector pcDNA3.1 (+) by DNA recombinant technology, then the restriction enzyme digestion and DNA sequencing were carried out to evaluate the recombinant. The pcDNA3.1 (+)-GAL and pcDNA3.1 (+) construct were transfected into immortalized rat astrocyte strain (IAST) by lipofectamine and the population of cells which stably integrated the construct was selected with 600 microg/mL G418. Individual clones were screened and expanded into clonal cell strains. Detection of Neo gene was used to validate the success of the transfection. Immunocytochemical staining, RT-PCR and radioimmunoassay were used to detect the expression and secretion level of GAL. The recombinant had been successfully constructed by restriction enzyme digestion and DNA sequencing. Detection of Neo gene showed that the pcDNA3.1 (+)-GAL and pcDNA3.1 (+) have been successfully transfected into IAST. After selection by using G418, IAST/GAL and IAST/Neo cell strains were obtained. IAST/GAL, IAST/Neo and IAST were immunostained positively for GAL, but the GAL average optical density of IAST/GAL was significantly higher than that of IAST/Neo and IAST (P< 0.01). The level of GAL mRNA expression and the supernatant concentration of GAL in cultured IAST/GAL were significantly higher than those of IAST and IAST/Neo (P<0.01), but no significant differences were found between the IAST and IAST/Neo (P>0.05). It was concluded that IAST/GAL strain was constructed successfully and it might provide a basis for the further study of pain therapy.
Astrocytes/cytology ; Astrocytes/*metabolism ; Cell Line, Transformed ; Cells, Cultured ; Galanin/*biosynthesis ; Galanin/genetics ; Genetic Vectors ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/genetics ; Transfection

Objective To construct lentivirus vector expressing antigene RNA and ferritin gene.Methods Intermediate plasmid pGC-FU-HF was constructed by transfecting lentivirus vector pGC-FU with heavy chain ferritin subunit gene.The target plasmid pGC-agRNA-HF was subsequently constructed by transfecting the intermediate plasmid with β-arrestin 2 antigene RNA.The NG108-15 cells were transfected with the target plasmid.The titre of lentivirus vector was measured by RT-PCR.The expression of antigene RNA and ferritin gene was determined by Western blot and RT-PCR.Results Lentivirus vector was successfully transfected with antigene RNA and ferritin gene.The titre of lentivirus vector was 2.00 × 109 TU/ml.The expression of β-arrestin2 protein was down-regulated and the expression of ferritin protein up-regulated in the NG108-15 cells after being transfected with the lentivirus vector.Conclusion Lentivirus vector expressing antigene RNA and ferritin gene has been successfully constructed.

Objective To detect the role of differentially expressed microRNA (miRNA) in human cholangiocarcinoma and explore their effects on apoptosis and invasiveness of cholangiocarcinoma.Methods The differential expression of miRNA in 3 cholangiocarcinoma patients was detected by miRNA array.The expressions of miR-200a,miR-200b,miR-200c,and miR-141 in human cholangiocarcinoma tissues and normal bile duct tissues were detected by real-time PCR.After transfection with miR-200b mimic,apoptosis and invasiveness of human cholangiocarcinoma cell line QBC939 was evaluated by Annexin-V-FITC dyeing and Transwell assay.Results Comparad with normal bile duct tissues,the number of differential miRNAs in cholangiocarcinoma was 21,including 15 up regulated and 6 down regulated.The expressions of miR200a,miR-200b,miR-200c,and miR-141 in human cholangiocarcinoma tissues were significantly lower than levels in normal bile duct tissues.The invasive ability of QBC939 was decreased after miR-200b mimic transfection.The apoptosis cell number of QBC939 was increased after miR-200b mimic transfection.Conclusion These results indicate that the expression of miRNA is different between cholangiocarcinoma and normal bile duct tissues.Moreover,miR-200a,miR-200b,miR-200c,and miR-141 are likely involved in the invasion and metastasis of cholangiocarcinoma and have potential as a diagnostic and prognostic marker.

OBJECTIVETo investigate the early clinical efficacy of induced membrane technique for reconstruction of large bone defects after debridement in adults with chronic osteomyelitis of limbs.

METHODSFrom March 2010 to March 2012,a total of 23 adult patients with chronic osteomyelitis of limbs were treated in our department. There were 15 males and 8 females, with a mean age 35.2 years old (ranged from 26 to 49 years old). Sixteen patients had open fracture history. According to the lesion site, there were 12 cases of tibia, 7 cases of femur, 3 cases of humerus, and 1 case of both radius and ulna. Among them, 19 patients had diseases in diaphysis and 4 patients in the metaphysis. The mean interval from infection to operation was 6.9 months (ranged from 4 to 13 months). All the patients were treated by using induced membrane technique. The follow-up evaluation included clinical complications, time of bone healing and limbs function. The Chinese version of SF-36 scores was used in the assessment of quality of life pre- and post-operation.

RESULTSThe average duration of follow-up was (27.6 ± 5.3) months (ranged from 18 to 43 months). Two patients had postoperative flap edge necrosis, 1 patient had superficial iliac incision infection, no obvious complications were recorded. Twenty patients obtained radiological union at a mean time of 4.6 months (ranged from 3 to 7 months). Among them, 16 patients treated with lower limbs surgery achieved full weight-bearing at about 5.2 months (ranged from 4 to 8 months) postoperatively. Four patients suffered from reinfection during follow-up, but 3 of them achieved complete bone healing after the second surgeries with induced membrane technique. At the final follow-up, there was a substantial improvement in each dimension scores and total scores of SF-36 as compared with those before surgery.

CONCLUSIONWhen treating with adult chronic osteomyelitis of limbs, the induced membrane technique can effectively reconstruct large bone defects after debridement, significantly shorten treatment cycle, provide satisfactory results with minimal complications, promote good recovery of limbs function and require relatively simple operation technique.

Adult ; Chronic Disease ; Extremities ; surgery ; Female ; Humans ; Male ; Middle Aged ; Osteomyelitis ; surgery ; Reconstructive Surgical Procedures ; methods

Objective To summarize the author' s clinical experiences in the diagnosis and treatment of urachal carcinoma.Methods Thirteen cases with urachal carcinoma from 1990-2011 at Beijing Friendship Hospital affiliated to Capital Medical University were retrospectively reviewed and analyzed.Results The most common complaint was hematuria;B ultrasonic or CT scan demonstrated a parenchyma or vesica between the bladder dome and abdominal wall.Nine patients underwent extensive pattical excision of the bladder,3 paitents underwent radical resection,and 4 patients received comprihensive chemotherapy.The five-year survival rate was 30.7％.Conclusion Extensive partial excision of the bladder is recommended for urachal carcinoma.

Objective To determine the efficacy of liver cancer resection combined with splenectomy for patients with hepatocellular carcinoma with hypersplenism.Methods Among 35 patients with hepatocellular cancer and hypersplenism treated from March 2004 to January 2006 at our hospital,12 patients accepted simultaneous liver cancer resection and splenectomy (the splenectomy group)and 23 only accepted liver cancer resection (the non-splenectomy group).The liver function,platelets and white blood cells were analyzed retrospectively.Results All the operations were successfully carried out.Within 1 week after operation,the white blood cell count increased from (3.2 ± 1.7) × 109/L to (8.5±-5.3) × 109/L,the platelet count increased from (52.6±23.7) × 109/L to (245.3±94.6) ×109/L(P＜0.01) in the group of patients with combined splenectomy,while little change occurred in the non-splenectomy group.The liver function in the splenectomy group recovered to the preoperational value within 1 week.Two years after operation,7 (58.3％) patients were still surviving in the splenectomy group and the mean tumor-free survival was (16.4 ± 4.3) months compared with (14.3 ±5.2) months in 10 (43.5％) patients in the non-splenectomy group,(P＜0.005).Conclusion Liver cancer resection combined with splenectomy was efficacious to hepatocellular cancer with hypersplenism.