【Objective】 To establish an enzyme-linked immunosorbent assay (ELISA) method for the determination of residual human coagulation factor Ⅺ in human prothrombin complex and validate the method. 【Methods】 Human factor Ⅺ was reacted with the capture antibody coated on the microtiter plate. After appropriate washing steps, biotinylated primary antibody was bound to the captured protein. Excess primary antibody was washed away and bound antibody was reacted with horseradish peroxidase conjugated streptavidin. TMB substrate was used for color development at 450 nm. The dilution reliability, accuracy, specificity, repeatability, intermediate precision, linearity, range and durability were verified. 【Results】 The verification results showed that the accuracy and specificity of this method met the experimental requirements, with an average recovery rate of 109.2% and RSD of 6.93%. The repeatability RSD was 6.78%, and the intermediate precision RSD was 6.75%, indicating good precision. The linear regression correlation coefficient of standard curve was 0.999 9, showing good accuracy and precision within the linear range. The durability was verified by the incubation time and the validity period of reagent kit opening. The results showed that the RSD of the incubation time change was 6.62%, indicating that the incubation time of this detection method was controlled between 28 to 32 minutes, and there was no significant impact on the results. The RSD of the detection results before and after the reagent kit was opened and stored under conditions for 7 days was 3.84%, indicating that the preservation of the reagent kit according to the conditions for 7 days after opening has no effect on the FⅪ detection results. Both indicated that the method had good durability. The dilution reliability results showed that there was a "hook" effect in the detection of FⅪ residue in human prothrombin complex, which could be solved by diluting 100 to 200 times. 【Conclusion】 This method can be used for the determination of FⅪ residues of human prothrombin complex in laboratory.

OBJECTIVE：To stud y the effects of sinapine thiocyanate （ST） on the proliferation ，epithelial mesenchymal transformation（EMT）and metastasis of human cutaneous squamous cell carcinoma SCL- 1 cells，and to investigate its possible mechanism. METHODS ：Human cutaneous squamous cell carcinoma SCL- 1 cells were divided into blank control group （0.1％ DMSO） and ST different concentration groups （5，10，20 μmol/L）. CCK- 8 assay，5-ethynyl-2′-deoxyuridine（EDU）test， scratch test and Transwell chamber invasion test were adopted to test the proliferation ，migration and invasion ability. The expression of N-cadherin and E-cadherin were detected by Western blot and immunofluorescence assay . Other SCL- 1 cells were collected and divided into blank control group （0.1％ DMSO），ST group （20 μmol/L），ST+NSC228155 group [ 20 μmol/L ST+100 μmol/L NSC228155（EGFR agonist ）] and ST+SC 79 group [ 20 μmol/L ST+20 μmol/L SC79（PI3K/Akt agonist ）]. The proliferation ，migration and invasion ability of SCL- 1 cells in each group were detected by CCK- 8 assay，scratch test and Transwell chamber invasion assay. The expression of epidermal growth factor receptor （EGFR），phosphatidylinositol 3 kinase（PI3K），phosphorylated phosphatidylinositol 3 kinase（p-PI3k），protein kinase B （Akt）and phosphorylated protein Akt （p-Akt）protein of cells in blank control group and ST different concentration groups（5，10，20 μmol/L）were determined by Western blot assay so as to validate the relationship between ST effect and EGFR/ PI3K/Akt signaling pathway. SCL- 1 cells and human normal skin fibroblasts cell WS 1 were divided into blank control group （0.1％ DMSO），ST group （20 μmol//L），ZD1839 group（positive control ，20 μmol//L，EGFR inhibitor ）and LY 294002 group（positive control，20 μmol//L，PI3K/Akt inhibitor ）. CCK- 8 assay was used to detect the cell proliferation in order to evaluate the cells cytotoxicity of ST. RESULTS ：Compared with blank control group ，the proliferation ，migration and invasion ability of SCL- 1 cells were significantly decreased in 5，10，20 μmol/L ST groups（P＜0.05）. Western blot and immunofluorescence assay showed that the expression of N-cadherin in SCL- 1 cells were decreased significantly in 5，10，20 μmol/L ST groups（P＜0.05），while the protein expression of E-cadherin was increased significantly （P＜0.05）；the protein expressions of EGFR ，p-PI3K and p-Akt were significantly decreased （P＜0.05）. Compared with ST group ，the proliferation ，migration and invasion ability of SCL- 1 cells were increased significantly in ST + NSC 228155 group and ST + SC 79 group （P＜0.05）. Compared with blank control group ，the proliferation ability of WS 1 cells had no significant change in ST group ，while the proliferation ability of SCL- 1 cells was decreased significantly （P＜0.05）；the proliferation ability of the two kinds of cells were decreased significantly in ZD 1839 group and LY 294002 group（P＜0.05）. Compared with ST group ，the proliferation ability of WS 1 cells was decreased significantly in ZD1839 group and LY 294002 group（P＜0.05），but there was no significant difference in the proliferation ability of SCL- 1 cells （P＞0.05）. CONCLUSIONS ：ST may inhibit the proliferation ，EMT and metastasis of SCL- 1 cells through inhibiting the activation of EGFR/PI 3K/Akt signaling pathway ，and its side effects are few.

Objective:To explore the effect of integrated application of multi-mode teaching in presentation-assimilation-discussion (PAD) class of medical microbiology.Methods:A total of 102 2017-batch clinical medical students of Hubei University of Medicine were collected, and the quasi-experimental study was adopted. The experimental class (50 students) was established by setting a PAD class, integrating the use of WeChat public account, Rain Classroom and WJX software during the teaching implement. And traditional mode of teaching was carried out in the control class (52 students). Moreover, the questionnaire survey and the test assessment were applied to compare the teaching effects of the two classes.Results:Questionnaire results showed that scores of indicators related to the teaching effectiveness in the experimental group were significantly higher than those in the control group ( P<0.01), and scores of tests in the experimental group were significantly better than those in the control group ( P<0.01). The t test was performed by SPSS 20 in the study. Conclusion:The integrated application of multi-mode teaching enriches the teaching mode of PAD class, which fully describes the student-centered concept, and increases students' initiative, participation, as well as students' knowledge mastery and innovation.

Objective:To explore the clinical effect of X-N advancement flap in repairing pressure ulcer on the buttock or back.Methods:From June 2018 to June 2019, 20 patients with grade Ⅳ pressure ulcers on the buttock or back were hospitalized and treated in the Department of Traumatology, Burns and Plastic Surgery of Fourth Affiliated Hospital of Guangxi Medical University, including 15 males and 5 females, aged 48-89 years. The area of the patient′s wound was 8 cm×5 cm-15 cm×12 cm after debridement, and all were repaired with the X-N advancement flap designed by the author. The flap was designed according to the direction of skin relaxation on both sides of the wound, and the skin was incised in X-shape and sutured in N-shape. The width and advancement distance of the flap were recorded, and the ratio of the advancement distance to the width of the flap was calculated. The flap survival, complication, and follow-up were observed and recorded.Results:The width of the flap was (5.9±1.2) cm, the advancement distance of the flap was (10.3±2.5) cm, and the ratio of the advancement distance to the width of the flap was 1.8±0.4. All the flaps survived, and none of the flaps had blood flow disorder. Local dehiscence occurred in the flap of one patient 1 week after surgery, which was healed after laying on the floating bed, strengthened care, and wound dressing change. The flap of one patient developed infection 5 days after surgery, which was healed after partial suture removal, smooth drainage, and replacement with sensitive antibiotics. The wounds of the remaining 18 patients were all cured. After 3 months of follow-up, the flaps survived well with good elasticity and texture.Conclusions:The X-N advancement flap can make the skin and soft tissue move forward effectively. It is simple and effective to repair pressure ulcers on the back or buttock of patients with this flap, which is worthy of clinical promotion and application.

Objective To analyze the risk factors of postoperative recurrence and metastasis of of BCLC stage A (BCLC-A) hepatocellular carcinoma (HCC).Methods Data of 210 cases of liver resection for HCC were retrospectively analyzed from Nov 2013 to June 2016.Multivariate logistic regression analysis was used to analyze the risk factors for postoperative recurrence.The cumulative survival rates were calculated using the Kaplan-Meier method and the diffrences of the related factors between various groups were analyzed by Log-rank test.Results Univariate analysis showed that alpha-fetoprotein (AFP),neutrophil-lymphocyte ratio (NLR),hepatitis B virus surface antigen (HBsAg) positive and intraoperative transfusion were associated with early recurrence and metastasis after hepatectomy (P ＜ 0.05).Multivariate analysis showed that AFP ＞ 400 ng/ml is an independent risk factor for early postoperative recurrence and metastasis (P =0.008).HBV DNA positive can affect the disease-free survival rate after HCC resection (P =0.030).Conclusion AFP is an independent risk factor for early postoperative recurrence and metastasis.Preoperative HBV DNA positive significantly decreases the disease-free survival rate in HCC patients.

Human infection with avian influenza A (H7N9) is an acute contagious respiratory disease. Acute respiratory distress syndrome (ARDS) is a common complication in patients with severe avian influenza A (H7N9), for whom mechanical ventilation (MV) is an important supportive method. A patient, suffered from severe avian influenza A (H7N9) and complicated with ARDS, was admitted to the Second Affiliated Hospital of Guizhou Medical University in January 2017. With very intensive care for oxygenation, respiration and consciousness, and monitoring, she was successfully cured by comprehensive managements, among which noninvasive mechanical ventilation (NIV) was the major respiratory support method. The result demonstrate that, in patients with conscious state, satisfied expectoration ability and relatively good cooperation, and with close observation of oxygenation and respiratory rate, NIV may be accepted as an effective method for patient with ARDS caused by severe avian influenza A (H7N9).

Objective To systematically review ICU-nurse lightly sedated patients, in order to provide evidence for sedation. Methods Databases were searched including The Cochrane Library, Joanna Briggs Institute Library, PubMed, EMbase, Ovid, ELSEVIER CMB, CNKI and WANFAN from inception to May 2018, to collect qualitative studies in the experience of ICU nurses lightly sedated patients. The quality of included studies was evaluated according to JBI Critical Appraisal Tool for qualitative studies in Australia. The results were integrated by integrating methods. Results A total of seven studies were included. Thirty-five complete findings were grouped according to their similarities to form six categories. These categories resulted in two synthesized findings: Despite the complexity and unknowns of care, critical care nurses were slowly accepting the fact and try to cope with; there were some barriers on the implementation of sedation, there should have a multi-party collaboration. Conclusions the heath management system should pay more attention to implementing light sedation strategies, ICU-nurses should be given support, guidance and encouragement.

Objective To summarize the experience of patient positing,port placements setting and robot cart docking in pediatric robot-assisted laparoscopic upper urinary tract operations.Methods From March 2017 to December 2017,140 robot-assisted laparoscopic upper urinary tract procedures were performed in our institution,including 110 cases of pyeloplasty,15 upper pole heminephroureterectomy,12 simple nephrectomy and 3 adrenalectomy.There were 103 males and 37 females with a range age from 1 month to 18 years.The assistant surgeon was adjacent to the instrument nurse,and patients were placed in a supine position with 60°-80° inclination and keep the legs low to the body.Room setup and patient positioning were similar to the traditional laparoscopic surgery.Semi-hidden incision technique was used in 140 patients:the camera port was placed umbilicus,two additional arm ports (one 5 mm and one 8 mm) were placed under direct vision,the 8 mm arm port was placed on the line of a Pfannenstiel incision and the 5 mm arm port was placed below the Xiphoid along the midline.Finally,a 3 or 5 mm assistant port was placed approximately 3 cm lateral to the inferior arm port,the line of a Pfannenstiel incision.Results The average time was (11.5 ± 3.2) min (10.5-16.5 min) from skin incision to robot cart docking completed.All surgeries were successfully completed without open conversion.One patient required an additional assist port for severe adhesion after the previously open surgery,there was no injury to other viscera.Average operative time was (146.9 ± 48.7)min (78-259 min) and average post-operative hospitalization time was (5.7 ± 1.4) d(4-10 d),respectively.There was no visual scar on abdominal 6 weeks postoperatively,and all parents made comments about their satisfaction with the cosmetic appearance.All operations got complete success at a mean follow up of 6 (1-9) months.Conclusions A good room setup,patient positioning and the semi-hidden incision technique port placements are maintaining the safety of the patient,avoiding compression injuries,allowing maximum mobility of the robotic arms,and facilitating a smooth and efficient surgery,and improving post-operative recovery.

One confirmed diagnosiscase of severe human infection by avian influenza H7N9 admitted to intensive care unit (ICU) of the Second Affiliated Hospital of Guizhou Medical University on January 12th, 2017 was reported. The patient was treated with the sepsis bundle, and recovered finally, including a series of comprehensive treatments, such as respiratory support, circulation support, antiviral, anti-inflammation, immunization enhancement, critical nursing, fluid management, nutritional support and treatment of complications. The critical patient was admitted on January 27th, and the treatment was successful. It has important significance to rescue the severe human infection from avian influenza H7N9 by the sepsis bundle.