Objective: To study the effect of HSVtk/GCV system on ascites tumors. Methods: Using the methods of modified XTT assay, animal experiment, transmission electron microscopy (TEM ) and flow cytometric ( FCM) assay. Results: The sensitivity of P388tk cell was about 37-fold than that of their patental P388 cell. Furthermore we found P388tk cells exhibited potent bystander killing. In vivo , the growth of tumors, which were produced by injecting P388tk cells or a mixture of 50% P388tk cells and 50% P388 cells into DBA/2 mice, was inhibited , furthermore mice survival periods were prolonged contrasted to control groups. Conclusion: HSVtk/GCV suicide gene system could effectively killed the HSVtk-gene-positive P388tk cells and nearby HSVtk-gene-negative cells by the bystander effect in vitro and in vivo.

Objective:To establish a mouse model with the main characteristics of asthma and to study the pathogenesis of its airway inflammation.Methods:Mice were given latex by the intraperitoneal sensitization/intranasal challenge.BALF histology were examined 24 hours following tracheal challenge;cytokines and serum total and specific IgE were examined in one month and the expression of IL-5?IFN-? were performed by ELISA ?RT-PCR.Results:After the latex challenge,in experimental group,the number of total BALF cells and the percentage of eosinophils elevated;the histological analysis showed the elevation of mucus production and goblet cell development within bronchial epithelium,spasm and constraction of bronchia ,marked peribronchial infiltration of eosinophils and lymphocytes; the elevation of total and specific IgE;reduction of IFN-? and increase of IL-5 in the BALF and lung.Conclusion:Latex sensitization and challenge can lead to airway eosinophilia,allergen-specific IgE induction and production of Th2 type cytokines, which were the main characteristics of asthma.

Objective To evaluate the value of neutrophil CD64 positive cells percentage(CD64%)detec-tion to stop using antibiotic in patients with severe pneumonia.Methods 60 accepted antibiotic therapy patients with severe pneumonia,in accordance with the random number table,were separated into observation group(n =30)and control group(n =30).Antibiotics were stopped according to CD64% in observation group,while it according to the clinical symptoms,the plasma level of white blood cell and C -reactive protein in control group.The main observation indexes included the days of antibiotics use,the length of Intensive Care Unit(ICU)stay,clinical efficacy and the case fatality rate.Results The days of antibiotics use in the observation group was (10.3 ±5.2)d,while it was (16.8 ± 5.8)d for patients in the control group,and it had significant difference(t =-4.570,P <0.01).The length of ICU stay in the observation group was shorter than that in the control group[(6.5 ±3.5)d vs (10.5 ±4.5)d],and it had significant difference(t =3.843,P <0.01).The clinical efficacy were 83.67% and 82.12%,and the case fatality rate were 9.68% and 10.24% in the observation group and control group,respectively,and both had no significant difference(P >0.05).Conclusion Stop using antibiotics according to the neutrophil CD64 % is safe,reliable,and can effectively reduce the excessive use of antibiotics and shorten the length of ICU stay in patients with severe pneumonia.

Objective To investigate the clinical significance of pulse high-volume hemofiltration (PHVHF) on expressions of Toll-like receptor (TLR) 2 and TLR4 mRNA in peripheral blood mononuclear cells (PBMCs) in patients with severe sepsis.Methods Forty patients with severe sepsis were divided into conventional treatment group (n =20) and PHVHF group (n =20) according to random number table.Another fifteen healthy volunteers served as controls.TLR2 and TLR4 mRNA expressions in PBMCs were detected using RT-PCR and plasma concentrations of TNF-α and IL-6 were measured using ELISA method before therapy and at 24 h,48 h and 72 h after therapy.Vital signs,BIL,Cr,BUN,Lac,PaO2/FiO2,acute physiology,chronic health evaluation Ⅱ (APPACHE Ⅱ),sequential organ failure assessment (SOFA) and prognosis were compared among the groups.Besides,complications associated with PHVHF therapy were monitored.Results Expressions of TLR2 and TLR4 mRNA in PBMCs and concentrations of TNF-α and IL-6 were significantly higher in patients with severe sepsis than in the controls (P ＜ 0.01).At 72 hours after therapy,PHVHF group showed significantly lower concentrations of TNF-α and IL-6 than those before therapy (P ＜ 0.01) as well as significantly lower expressions of TLR2 and TLR4 mRNA in PBMCs than those in conventional treatment group (P ＜ 0.O1).However,no significant decline in the levels of TNF-α and IL-6 and the expressions of TLR2 and TLR4 mRNA in PBMCs were shown in conventional treatment group after therapy.At 72 hours after therapy,PHVHF group showed significant increases of MAP and PaO2/FiO2 and significant decreases of Cr,BUN,Lac,APACHE Ⅱ and SOFA as compared to those before therapy (P ＜ 0.05).Moreover,the differences of MAP,PaO2/FiO2,Cr,BUN,Lac,APACHE Ⅱ and SOFA were statistically significant between and conventional treatment group at 72 hours after therapy (P ＜ 0.05).Conclusion PHVHF achieves a reduced systemic inflammatory response,improved major organ functions,shortened length of stay in ICU and down-regulated TLR2 and TLR4 expressions in PBMCs that may be a novel mechanism of PHVHF in treatment of severe sepsis.

This paper introduces a brand-new three-way trachea tube reinforced by steel.It has a three-way junction.The main tube has the function of ventilation and connection,while the side tube which can be connected to oxygen-breath tube is used for the autonomic breath of the patient or for nurses to draw secretions.Furthermore,the steel wire is inserted in the tubes in case it is snapped.

Objective To analyze indications,complications and outcomes of amputation.Methods A total of 15 patients undergone amputation in field or at tent hospital were collected for analy-zing injury severity,place where amputation was done,whether open or closed amputation and stitch re-moval time. Results There were 9 males and 6 females.at an average age of 32 years(11-51years).There were 16 amputations including Gustilo IIIB in 2 patients, Gustilo IIIC in 9 and Tscheme Ⅲin 5 according to Gustiln classification or Tscheme classification.Four patients who received amputation in field or at tent hospital developed infection and had to receive amputation again at a higher level on the limb and drainage of open wounds because of a higher infection rate due to the amputation location.Ten patients received first amputation at higher levels with open wound at station hospital but only 2 manifested infected incision.High level amputation with one stage closure was done in 1 patient who was infected and suppurated after operation and even developed bacteremia. Conclusions Infection rate following am-putation 4n field and tent clinics is rather higher,so secondary open amputations should be performed at a higher level as soon as possible.One-time and high-level open amputation plays an important role in treat-ment of severe lower limb injuries following earthquake.

Objective:To construct a prognosis associated micro RNA(miRNA) prediction model based on bioinformatics analysis and evaluate its application value in pancreatic cancer patients.Methods:The retrospective cohort study was conducted. The clinicopathological data of 171 pancreatic cancer patients from the Cancer Genome Atlas (TCGA) (https: //cancergenome.nih.gov/) between establishment of database and September 2017 were collected. There were 93 males and 78 females, aged from 35 to 88 years, with a median age of 65 years. Of the 171 patients, 64 had complete clinicopathological data. Patients were allocated into training dataset consisting of 123 patients and validation dataset consisting of 48 patients using the random sampling method, with a ratio of 7∶3. The training dataset was used to construct a prediction model, and the validation dataset was used to evaluate performance of the prediction model. Nine pairs of miRNA sequencing data (GSE41372) of pancreatic cancer and adjacent tissues were downloaded from Gene Expression Omnibus database. The candidate miRNAs were selected from differentially expressed miRNAs in pancreatic cancer and adjacent tissues for LASSO-COX regression analysis based on the patients of training dataset. A prognosis associated miRNA prediction model was constructed upon survival associated miRNAs which were selected from candidate differentially expressed miRNAs. The performance of prognosis associated miRNA prediction model was validated in training dataset and validation dataset, the accuracy of model was evaluated using the area under curve (AUC) of the receiver operating characteristic curves and the efficiency was evaluated using the consistency index (C-index). Observation indicarors: (1) survival of patients; (2) screening results of differentially expressed miRNAs; (3) construction of prognosis associated miRNA model; (4) validation of prognosis associated miRNA model; (5) comparison of clinicopathological factors in pancreatic cancer patients; (6) analysis of factors for prognosis of pancreatic cancer patients; (7) comparison of prediction performance between prognosis associated miRNA model and the eighth edition TNM staging. Measurement data with normal distribution were represented as Mean± SD, comparison between groups was analyzed by the student- t test, and comparison between multiple groups was analyzed by the AVONA. Measurement data with skewed data were represented as M (range), and comparison between groups was analyzed using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Ordinal data were analyzed using the rank sum test. Correlation analysis was conducted based on count data to mine the correlation between prognosis associated miRNA model and clinicopathological factors. COX univariate analysis and multivariate analysis were applied to evaluate correlation with the results described as hazard ratio ( HR) and 95% confidence interval ( CI). HR<1 indicated the factor as a protective factor, HR>1 indicated the factor as a risk factor, and HR equal to 1 indicated no influence on survival. The Kaplan-Meier method was used to draw survival curve and calculate survival rates, and the Log-rank test was used for survival analysis. Results:(1) Survival of patients: 123 patients in the training dataset were followed up for 31-2 141 days, with a median follow-up time of 449 days. The 3- and 5-year survival rates were 16.67% and 8.06%. Forty-eight patients in the validation dataset were followed up for 41-2 182 days, with a median follow-up time of 457 days. The 3- and 5-year survival rates were 15.63% and 9.68%. There was no significant difference in the 3- or 5-year survival rates between the two groups ( χ2=0.017, 0.068, P>0.05). (2) Screening results of differentially expressed miRNAs. Results of bioinformatics analysis showed that 102 candidate differentially expressed miRNAs were selected, of which 63 were up-regulated in tumor tissues while 39 were down-regulated. (3) Construction of prognosis associated miRNA model: of the 102 candidate differentially expressed miRNAs, 5 survival associated miRNAs were selected, including miR-21, miR-125a-5p, miR-744, miR-374b, miR-664. The differential expression patterns of pancreatic cancer to adjacent tissues were up-regulation, up-regulation, down-regulation, up-regulation, and down-regulation, respectively, with the fold change of 4.00, 3.43, 3.85, 2.62, and 2.35. A prognostic expression equation constructed based on 5 survival associated miRNAs = 0.454×miR-21 expression level-0.492×miR-125a-5p expression level-0.49×miR-744 expression level-0.419×miR-374b expression level-0.036×miR-664 expression level. (4) Validation of prognosis associated miRNA model: The C-index of prognosis associated miRNA model was 0.643 and 0.642 for the training dataset and validation dataset, respectively. (5) Comparison of clinicopathological factors in pancreatic cancer patients: results of COX analysis showed that the prognosis associated miRNA model was highly related with pathological T stage and location of pancreatic cancer ( Z=45.481, χ2=10.176, P<0.05). (6) Analysis of factors for prognosis of pancreatic cancer patients: results of univariate analysis showed that pathological N stage, radiotherapy, molecular targeted therapy, score of prognosis associated miRNA model were related factors for prognosis pf pancreatic cancer patients ( HR=2.471, 0.290, 0.172, 2.001, 95% CI: 1.012-6.032, 0.101-0.833, 0.082-0.364, 1.371-2.922, P<0.05). Results of multivariate analysis showed that molecular targeted therapy was an independent protective factor for prognosis of pancreatic cancer patients ( HR=0.261, 95% CI: 0.116-0.588, P<0.05) and score of prognosis associated miRNA model≥1.16 was an independent risk factor for prognosis of pancreatic cancer patients ( HR=1.608, 95% CI: 1.091-2.369, P<0.05). (7) Comparison of prediction performance between prognosis associated miRNA model and the eighth edition TNM staging: in the training dataset, there was a significant difference in the prediction probability for 3- and 5-year survival of pancreatic cancer patients between prognosis associated miRNA model and the eighth edition TNM staging ( Z=-1.671, -1.867, P<0.05). The AUC of the prognosis associated miRNA model and the eight edition TNM staging for 3- and 5-year survival prediction was 0.797, 0.935 and 0.737 , 0.703, with the 95% CI of 0.622-0.972, 0.828-1.042 and 0.571-0.904 , 0.456-0.951. The C-index was 0.643 and 0.534. In the validation dataset, there was a significant difference in the prediction probability for 3- and 5-year survival of pancreatic cancer patients between prognosis associated miRNA model and the eighth edition TNM staging ( Z=-1.729, -1.923, P<0.05). The AUC of the prognosis associated miRNA model and the eight edition TNM staging was 0.750, 0.873 and 0.721 , 0.703, with the 95% CI of 0.553-0.948, 0.720-1.025 and 0.553-0.889, 0.456-0.950, respectively. The C-index was 0.642 and 0.544. Conclusions:A prognosis associated miRNA prediction model can be constructed based on 5 survival associated miRNAs in pancreatic cancer patients, as a complementation to current TNM staging and other clinicopathological parameters, which provides individual and accurate prediction of survival for reference in the clinical treatment.

Objective:To use three-dimensional (3D) scanning to measure the preoperative and postoperative nasal parameters of the patients received rhinoplasty with ear cartilage and silicone prosthesis, and to evaluate the clinical effect of the surgery.Methods:Sixteen female patients with an average age of 28.3 years, ranged from 21 to 35 years, received rhinoplasty with ear cartilage and silicone prosthesis in Wuhan Tongji Hospital from June 2018 to February 2019. Preoperative and postoperative 3D scanning was performed to measure nasal parameters, including linear length, angle, and proportional index.Results:All patients were satisfied with the postoperative outcomes. The postoperative nasal length, nasal height, and nasal depth increased significantly, and the postoperative nasal width and nasal tip width decreased. The postoperative nasolabial angle and nasofrontal angle were statistically improved, while the preoperative and postoperative data of columellar facial angle, nasal tip angle, and nasal column-lobular angle were not significantly different. The ratios of nasal depth and nasal width, nasal index and nasal tip protrusion were improved after surgery, while the postoperative ratios of nasal columella and nasal lobules length were not statistically improved.Conclusions:The 3D scanning allows for comprehensive and accurate measurement of the nasal parameters. The rhinoplasty with ear cartilage and silicone prosthesis is more effective in improving the overall contour of the nose, but less effective in improving the aesthetics of the nasal tip.

Objective To explore the key points of surgical techniques in individualized breast augmentation via inframammary fold (IMF) incision.Methods Fifty-six patients of our department from March 2014 to March 2017 in this group were followed up for 3-18 months,with average 12 months.According to the patients' characteristics to determine the size and dimensions of the implant and the exact position of new IMF,via IMF incision we inserted an implant part under the pectoralis major muscle and part posterior to the breast after precisely stripping and forming the implant pocket,which were the dual-plane augmentation mammoplasty.Results Compared with traditional endoscope-assisted breast augmentation via axillary incision,the breast augmentation via inframammary fold incision was more accurate,the operation time was shortened by an average of one hour,the intraoperative hemorrhage decreased by about 20 ml on average,the postoperative drainage fluid was reduced by about 70 ml on average,the hospitalization time was shortened by about 3 days,the postoperative breast shape was naturally upright,and the incision was hidden at the new inframammary fold.No hematoma,infection,significant hypertrophic scar,capsule contracture and other complications were noted.Conclusions The individualized dual-plane augmentation mammoplasty via IMF incision can hide the incision in the inframammary fold.The operation under direct vision and no use of special equipments make it an ideal breast augmentation surgery.

Objective: To explore the regulatory mechanism of E2F1 transcription factor on M2 macrophages in full-thickness skin defect wounds of mice. Methods: E2F1 gene knockout heterozygotes C57BL/6 mice and wild-type C57BL/6 mice were introduced and self-reproduced. Two weeks after birth, E2F1 gene knockout homozygotes mice and wild-type mice were identified by polymerase chain reaction (PCR). Twelve identified 6-8 weeks old male E2F1 gene knockout homozygotes C57BL/6 mice and wild-type C57BL/6 mice were selected respectively according to the random number table and set as E2F1 gene knockout group and wild-type group. A full-thickness skin defect wound was made on the back of each mouse. On post injury day (PID) 2 and 7, 6 mice in each group were selected according to the random number table and sacrificed, and the wound tissue was excised. The expression of CD68 and CD206 double positive M2 macrophages was observed by immunofluorescence method, and the percentage of CD206 positive cells was calculated. The protein expression of CD206 was detected by Western blotting. The mRNA expression of arginase 1 was detected by real-time fluorescent quantitative reverse transcription PCR (RT-PCR). Wound tissue specimens of the two groups on PID 7 were obtained, and the protein and mRNA expressions of peroxisome proliferator-activated receptor gamma (PPAR-γ) were detected by Western blotting and real-time fluorescent quantitative RT-PCR respectively. The above-mentioned experiments were repeated four times. Three specimens of wound tissue of mice in wild-type group on PID 7 were obtained to detect the relationship between E2F1 and PPAR-γ by co-immunoprecipitation and Western blotting, and this experiment was repeated two times. Data were processed with unpaired t test. Results: The size of PCR products of E2F1 gene knockout homozygotes C57BL/6 mice and wild-type C57BL/6 mice were 227 and 172 bp respectively, which were the same as those of the designed DNA fragments. On PID 2 and 7, the number of CD68 and CD206 double positive M2 macrophages in the wound tissue of mice in E2F1 gene knockout group was more than that of wild-type group, and the percentages of CD206 positive cells in the wound tissue of mice in E2F1 gene knockout group were (0.234±0.032)% and (0.584±0.023)% respectively, which were significantly higher than (0.129±0.017)% and (0.282±0.071)% of wild-type group (t=3.29, 3.54, P<0.05). On PID 2 and 7, the protein expression of CD206 in the wound tissue of mice in E2F1 gene knockout group were 1.00±0.23 and 1.63±0.26 respectively, which were significantly higher than 0.43±0.06 and 0.97±0.08 of wild-type group (t=2.41, 2.45, P<0.05). On PID 2 and 7, the mRNA expressions of arginase 1 in the wound tissue of mice in E2F1 gene knockout group were 0.482±0.105 and 0.195±0.031 respectively, which were significantly higher than 0.163±0.026 and 0.108±0.017 of wild-type group (t=3.04, 2.86, P<0.05). On PID 7, the protein and mRNA expressions of PPAR-γ in the wound tissue of mice in E2F1 gene knockout group were 0.61±0.12 and 0.51±0.13 respectively, which were significantly higher than 0.20±0.04 and 0.20±0.04 of wild-type group (t=3.36, 2.86, P<0.05). On PID 7, detection of the wound tissue of mice in wild-type group showed that PPAR-γ had unidirectional effect on E2F1. Conclusions E2F1 transcription factor affects the polarization of M2 macrophages by inhibiting the expression of PPAR-γ, thereby inhibiting the healing process of full-thickness skin defect wounds in mice.