0.05). Conclusion:The stress reaction to human injury stimulation are inhibited effectively by propofol - fentanyl intraveous anesthesia

Objective To construct the recombinant adeno-associated virus vector with human preproenkephalin gene (rAAV-hPPE). Methods The human preproenkephalin (hPPE) gene was cloned into the adeno-associated virus (AAV) vector plasmid pSNAV which contained neo expression box. The recombinant pSNAV-hPPE was then transfected into BHK cells using lipofectamine?2000. The G418-resistant cells, BHK / SH1, were obtained. The BHK / SH1 cells were infected with HSV1-rc /△UL2 which has the function of packaging the recombinant AAV(rAAV) . After purification, the construction of rAAV-hPPE was achieved.Results The construction of pSNAV-hPPE was confirmed by digestion with restriction enzyme. Southeon bolting was used to detect the virus liters (2.5 ? 1212 v.g /ml) .Conclusion This rAAV-hPPE virus vector with high liter and strong infectivity can be used in transgenic analgesic research.

Objective To study the effects of profound hemodilution with 6% HES (200/0.5) on coagulation.Methods Ten male new Zealand long-ear rabbits weighing (2.43 ?0.19) kg were anesthetized with intravenous thiopentone 8 mg-1 and tracheotomized and mechanically ventilated (VT = 15 ml-1 , RR = 24 bpm) . Anesthesia was maintained with intravenous infusion of thiopentone and succinylcholine. Femoral artery and vein were cannulated for BP and CVP monitoring. Hemodilution was performed in 6 steps at 30 min intervals. Blood was withdrawn from artery and simultaneously replaced by intravenous infusion of equal volume of 6% HES until Hct was 5 % -8 % . Blood samples were taken before hemodilution and 30 min after each step of hemodilution for determination of coagulation function curve using Sonoclot coagulation and platelet function analyzer. Results When Hct was 25%-30% there was no significant difference in coagulation function before and after hemodilution. When Hct was 15%-20% there was significant difference in ACT, TTP, clot rate and MCS before and after hemodilution but the normal coagulation process was not affected. When Hct

Objective To investigate the expressions and correlations of Kiss-1 and matrix metalloproteinases-9 (MMP9) proteins in the colorectal cancer (CRC) tissues of patients with synchronous colorectal liver metastasis (SCRLM),and the association with the clinicopathologic factors and prognosis of patients.Methods The retrospective case-control study was adopted.The clinicopathological data of 96 patients with SCRLM and 69 patients with CRC and no metastasis who were admitted to the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University from January 2000 to May 2013 were collected.The 96 CRC tissues and 50 adjacent normal tissues (distance from resection margin ≥ 5 cm) were collected from 96 patients with SCRLM,and 69 CRC tissues were collected from 69 patients with CRC and no metastasis.Expressions of Kiss-1 and MMP9 protein were detected by immunohistochemistry (IHC).The follow-up of outpatient examination and telephone interview was performed to detect survival of patients till August 2014.Comparison of count data and correlation between expressions of Kiss-1 or MMP9 protein and clinicopathological factors were analyzed by the chi-square test and Fisher exact probability.Survival curve was drawn using the Kaplan-Meier method,and survival analysis was done using the Log-rank test.Correlation analysis was done by the Pearson correlation.Results Expression of Kiss-1 protein was located in the cytoplasm of tissue cells.The positive expression rates of Kiss-1 protein in CRC tissues of patients with SCRLM and with CRC and no metastasis and in adjacent normal tissues were 24.0％ (23/96),43.5％ (30/69) and 52.0％ (26/50),respectively,with a significant difference among the 3 tissues (x2 =14.307,P ＜ 0.05) and no significant difference between CRC tissues of patients with CRC and no metastasis and patients with SCRLM (x2 =0.845,P ＞ 0.05).The positive expression rate of Kiss-1 protein in CRC tissues of patients with SCRLM was significantly different from that in CRC tissues of patients with CRC and no metastasis and in adjacent normal tissues (x2 =0.702,11.594,P ＜ 0.05).Expression of MMP9 protein was located in the cytoplasm of tissue cells.The positive expression rates of MMP9 protein in CRC tissues of patients with SCRLM and with CRC and no metastasis and in adjacent normal tissues were 67.7 ％ (65/96),62.3 ％ (43/69) and 36.0％ (18/50),respectively,with a significant difference among the 3 tissues (x2=14.203,P ＜0.05) and no significant difference between CRC tissues of patients with CRC and no metastasis and patients with SCRLM (x2=8.038,P ＞ 0.05).The positive expression rate of MMP9 protein in CRC tissues of patients with SCRLM was significantly different from that in CRC tissues of patients with CRC and no metastasis and in adjacent normal tissues (x2 =13.475,13.475,P ＜ 0.05).The positive expression rates of Kiss-1 protein in CRC tissues of patients with SCRLM were 66.7％,21.9％ and 17.6％ in the high-,mederate-and low-differentiated tumor,50.0％,28.6％ and 17.5％ in the muscular layer of tumor invasion,outside of serosa and serosal layer,44.0％ and 16.9％ in patients with and without lymph node metastasis,respectively,showing significant differences among the tumor differentiation degree,depth of tumor invasion and lymph node metastasis (x2=6.546,6.172,7.453,P ＜0.05).The positive expression rates of MMP9 protein in CRC tissues of patients with SCRLM were 25.0％,66.7％ and 76.2％ in the muscular layer of tumor invasion,outside of serosa and serosal layer,44.0％ and 76.1％ in patients with and without lymph node metastasis,respectively,showing significant differences between the depth of tumor invasion and lymph node metastasis (x2 =12.094,8.690,P ＜ 0.05).All the 96 patients with SCRLM were followed up for a median time of 68 months (range,12-176 months).The median overall survival time,median tumor-free survival time,5-year cumulative survival rate and 5-year tumor-free survival rate were 31 months,26 months,69.6％ and 26.1％ in patients with SCRLM and positive expression of Kiss-1 protein and 26 months,19 months,24.7％ and 12.3％ in patients with SCRLM and negative expression of Kiss-1 protein,respectively,showing significant differences between the overall survival and tumor-free survival (x2=16.578,14.436,P ＜ 0.05).The median overall survival time,median tumor-free survival time,5-year cumulative survival rate and 5-year tumor-free survival rate were 31 months,19 months,24.6％ and 12.3％ in patients with SCRLM and positive expression of MMP9 protein and 28 months,16 months,58.1％ and 22.6％ in patients with SCRLM and negative expression of MMP9 protein,respectively,showing significant differences between the overall survival and tumor-free survival (x2=14.073,8.532,P ＜0.05).Of 96 patients with SCRLM,there were 23 patients with positive expression of Kiss-1 protein (10 with positive expression of MMP9 protein and 13 with negative expression of MMP9 protein) and 73 with negative expression of Kiss-1 protein (55 with positive expression of MMP9 protein and 18 with negative expression of MMP9 protein),with a negative correlation between expressions of Kiss-1 protein and MMP9 protein (r =-0.291,P ＜ 0.05).Conclusions The reduced expression of Kiss-1 protein and elevated expression of MMP9 protein are closely associated with invasion and metastasis of CRC and prognosis of patients.A combination detection of Kiss-1 and MMP9 proteins is expected to become a marker for predicting the prognosis of patients with SCRLM based on the negative correlation between them.

Objective To investigate the effects of simvastatin on the expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) induced by oxidized low-density lipoprotein (ox-LDL) or Glucose in U937 macrophages, and explore the role of NF-κB in modulating of LOX-1 expression. Methods U937 macrophages were treated with PMA to induce differentiation, which were co-cultured with 50 mg/L ox-LDL or/and 25 mmol/L glucose. Pyrrolidine dithiocarbamate (PDTC) and simvastatin (1 μmol/L or 10 μmol/L) were used to treat cells. The expression of LOX-1 protein and NF-κB ac- tivity were detected by enzyme-linked immunosorbent assay technology. The expression of LOX-1 mRNA was measured by RT-PCR. Results The expression of LOX-1 was up regulated by ox-LDL, glucose and combination of both. The inhibitor of NF-κB PDTC suppressed this up-regulation. Simvastatin suppressed the expression of LOX-1 induced by ox-LDL, and showed a significant effect in the higher concentration. There was no significant effect of simvastatin on the expression of LOX-1 induced by glucose. The variation of NF-κB activity was similar to that of LOX-1 expression. Conclusion Simvas- tatin suppressed the expression of LOX-1 induced by ox-LDL, while no significant effect on the expression of LOX-1 in- duced by glucose. The expression and regulation of LOX-1 were related with NF-κB pathway.

OBJECTIVETo explore the clinical effects of minor bone anchors and palmaris longus tendon graft in treating chronic mallet fingers deformity.

METHODSFrom January 2008 to June 2013, 26 patients with chronic mallet fingers deformity were treated with minor bone anchors and palmaris longus tendon graft. There were 18 males and 8 females, aged from 18 to 52 years old with an average of (32.0±1.3) years. Among them, 8 cases caused by machine injury, 6 cases by fall injury, 6 cases by sprain from fight, 4 cases by tendon spontaneous rupture, 2 cases by knife trauma. There was no tendon attachment of extensor tendon check in 16 cases, and with 0.3 to 0.5 cm tendon attachment in 10 cases. All patients had the flexion deformity and the disability of dorsiflexion activity. During operation, the distal interphalangeal joint was fixed in 10° to 20° dorsiflexion by a Kirshner wire, the minor bone anchor was used to reconstruct the extensor tendon insertion, the palmaris longus tendon slice was transplanted the decayed area of extensor tendon insertion. Four weeks postoperatively, the Kirshner wire was removed and the plaster external fixation was used, and the patient began function exercises. Postoperative complications were observed and fingers functions were assessed according to Dargan standard.

RESULTSThe patients were followed up from 6 to 14 months with an average of (5.0±0.3) months. Wound superficial infection occurred in 2 cases, the skin pressure ulcer in 2 cases, joint activities disability in 1 case; these symptoms got improvement after symptomatic treatment. Traumatic arthritis occurred in 2 cases, 1 case was improved after treatment, and 1 case had chronic pain for a long time. No internal fixation loosening or breakage and tendon rupture were found. According to Dargan standard to evaluate the finger function, 17 cases got excellent results, 8 good, and 1 poor.

CONCLUSIONIt is an effective way to treat the chronic mallet finger deformity using minor bone anchors and palmaris longus tendon graft, and the method has advantages of reliable fixation, easy operation, satisfactory effect and less complication.

Adolescent ; Adult ; Female ; Finger Injuries ; surgery ; Fracture Fixation, Internal ; Hand Deformities, Acquired ; surgery ; Humans ; Male ; Middle Aged ; Suture Anchors ; Tendon Transfer

Adult ; Choanal Atresia ; etiology ; therapy ; Constriction, Pathologic ; etiology ; Humans ; Male ; Nasopharyngeal Diseases ; etiology ; therapy ; Otorhinolaryngologic Surgical Procedures ; adverse effects ; Palate ; surgery ; Postoperative Complications ; therapy ; Uvula ; surgery

Anemia, Aplastic ; therapy ; Cytomegalovirus Infections ; etiology ; Fatal Outcome ; Humans ; Immunotherapy ; adverse effects ; Male ; Pneumonia, Viral ; etiology ; Young Adult

ADAM Proteins ; blood ; immunology ; ADAMTS13 Protein ; Adult ; Female ; Humans ; Male ; Middle Aged ; Plasma Exchange ; Purpura, Thrombotic Thrombocytopenic ; blood ; pathology ; therapy

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