Objective: Based on the central-composite design (CCD), the genetic neural network (GNN) and genetic algorithm (GA) were applied to optimize the microwave extraction conditions of astragalus saponins. Methods: The HPLC fingerprint of astragaloside was constructed, and seven components (astragaloside I—V, isoastragaloside I, II) were selected to calculate the comprehensive score by the entropy weight method. On the basis of single factor experiment, CCD was used to designed the experimental condition. The quantitative relationship between extraction conditions and comprehensive score was established by GNN, and the optimal microwave extraction parameters of astragalus saponins were optimized by GA. Results: The optimal extraction conditions were obtained by GA-GNN. The extraction time was 260 s, the extraction power was 695 W, the ethanol content was 50%, the ratio of material to liquid was 21.5, and the comprehensive score of seven astragalosides was 1 432.584. Meanwhile, the optimal extraction conditions and comprehensive evaluation scores obtained were by response surface methodology (RSM). The extraction time was 190 s, the extraction power was 880 W, the ethanol content was 70%, the ratio of material to liquid was 18.5, and the comprehensive scores of seven astragaloside were 1 066.236. The experimental results showed that the extraction conditions obtained by GA-GNN can effectively increase the comprehensive score. Conclusion: It is feasible to construct a mathematical model between astragaloside components and microwave extraction conditions by using entropy weight method combined with GNN, which can provide a new scientific method for optimizing the extraction, separation, and purification of effective components of traditional Chinese medicine.

Objective To explore the changes in serum plasminogen activator inhibitor 1(PAI-1),tissue factor(TF) and antithrombin Ⅲ(ATⅢ) in early period of severe limb injury,and their relationship with the occurrence of traumatic pre-DIC and DIC after trauma.Methods Thirty-five patients were divided into severe injury group(AIS score ≥3,20 cases),minor injury group(15 cases),and 10 healthy subjects served as control.The 35 patients with injury were divided again into pre-DIC group(10 cases),DIC group(3 cases),and others(22 cases).Fasting peripheral venous blood was collected on day 1,3,6 from the patients and healthy subjects.The changes in TF,ATⅢ and PAI-1 levels were detected and statistically analyzed.Results The PAI-1 levels were higher in minor injury group and severe injury group than that in control group on day 1(P

Objective To explore the correlation between the strength of trunk flexors and extensors obtained by isometric and isokinetic concentric tests. Methods The strength of trunk extensors and flexors were measured by isometric and isokinetic concentric tests during trunk flexion and extension in 50 healthy subjects, and the peak torque of flexion and extension (FPT and EPT) and the ratios of FPT/EPT were analyzed with correlation and regression analysis. Results The FPT and EPT of trunk flexors and extensors were positively correlated (r_F=0.827,P0.05, respectively), the regression equations of FPT and EPT were y^=8.158+1.097X and y^=-12.375+0.781X, respectively. Conclusion The isometric test of the trunk extensors and flexors could assess accurately the changes of the trunk muscle strength without aggravating pain and diseases. It is more reasonable to evaluate the stability of trunk with the FPT/EPT ratio at neutral position.

Objective To explore the related factors with skin flap necrosis,we concluded the cases of patients with skin defects after free flap plantation.Methods From 2001 to 2016,188 cases about 20 influencing factors were analyzed (The characteristics of patients:age,sex,smoke,diabetes,high blood pressure;Preoperative factors:injured sections,injured causes,preoperative wound infection,preoperative wound osteomyelitis,the time from injury to operation;Intraoperative factors:operator,operation time,anesthesia time,intraoperative rehydration fluids,the way of vascular anastomosis,the number of venous anastomosis,the area of flap;Postoperative factors:flap hematoma,flap infection,vascular crisis) and multivariate logistic regression analysis was used to analyze the relationship between these risk factors and flap necrosis.Results All 188 cases were treated with free anterolateral thigh flap to repair soft tissue defect and it was revealed that the 174 cases were successful (92.55％) and 23 cases were occured vascular crisis (12.23％),8 cases were arterial crisis,11 cases were vein crisis,4 cases were ateriovenous crisis.After the treatment,the rescue was successful in 5 cases (38.46％).After the analysis we made the conclusion that the number of venous anastomoses,flap hematoma and vascular crisis were related with the skin flap necrosis.Conclusion The number of venous anastomose (≥2) will increase blood return to make the flap easier to survive.Intraoperative stanching and drainage tube placement work will reduce the skin flap hematoma as a result of reducing the skin flap necrosis.Artery and venous crisis handled in time,can enhance the survival rate of flap.

Objective To evaluate the concept of artery first and total mesopancreatic excision in radical resection of pancreatic head carcinoma through both anterior and posterior approaches.Method The anterior approach was to identify the superior mesenteric artery (SMA) and the posterior approach to confirm the possibility of negative margin at the origin of SMA,on the posterio-lateral vascular wall of superior mesenteric vein (SMV) and the supposed posterior of the mesopancreas.The resection scope were with the celiac trunk and common hepatic artery as the upper boarder,the SMA as the left boarder,the inferior mesenteric vein (IMA) level as the lower boarder,to achieve a complete mesopancreatic excision,namely the en bloc resection of all the involved nerve,the lymph tissue and vascular tissue along the right side of the axial composed by SMA and celiac trunk.Results Of the 15 patients,11 had radical Whipple procedure,among which 2 had a combining SMV resection and reconstruction.1 case suffered from delayed gastric emptying and 2 cases from bile leakage.There was no mortality.The postoperative pathology reported carcinoma in all 11 cases,with duodenum and low bile duct involved in 4 cases,with the duodenum involved in 6 cases,no surrounding tissue involvement was identified in 1 case.Nerve involvement was found in 7 (7/11),vascular involvement in 10 (10/11),and lymphnode metastasis was (2.5 ± 3.8/12.9 ± 4.9).Conclusions The radical resection of pancreatic head carcinoma using the concept of artery first and the total mesopancreatic excision is helpful for an early evaluation of the possibility of radical resection and guarantees negative margins.

AIM: To analyze the difference of endonuclease domain containing 1 (ENDOD1) expression be-tween benign prostatic hyperplasia ( BPH ) tissues and prostate cancer ( PCa ) tissues and to investigate the effect of ENDOD1 on the biological function of human prostate cancer cells .METHODS: The BPH samples ( n=20 ) and PCa samples (n=21) were processed and analyzed according to the instruction of immunohistochemical (IHC) staining.The mRNA and protein levels of ENDOD 1 in the normal prostate epithelial cells and prostate cancer cells were evaluated by RT -qPCR and Western blot , respectively .The recombinant plasmids pCMV-N-Flag-ENDOD1 was constructed and was trans-fected into the human prostate cancer cells .The proliferation , apoptosis , migration and invasion abilities of the prostate cancer cells were evaluated by MTT assay , flow cytometry, Transwell migration and Matrigel invasion assays , respectively. RESULTS:The analysis of variance of the immunoreactivity score showed that PCa tissues with high Gleason score dis -played significantly lower ENDOD1expression than that with low Gleason score and BPH (P<0.05).The expression of ENDOD1 at mRNA and protein levels in PC3 cells and DU145 cells was significantly lower than that in the LNCap cells (P<0.05).The proliferation of DU145 transfected with ENDOD1 was inhibited.The flow cytometry indicated that ENDOD1 over-expression in the DU145 cells resulted in a notable increase in G0/G1 phase arrest (P<0.05), but the ap-optotic rates showed no statistical difference .The results of Transwell assay showed that migration and invasion abilities of the cells were also inhibited after transfection with over-expressing ENDOD1 plasmid (P<0.05).CONCLUSION: The expression of ENDOD1 significantly decreased in prostate cancer with high Gleaon score .Meanwhile, the ENDOD1 is spe-cifically down-regulated in androgen independent prostate cancer (AIPC) cell lines.Over-expression of ENDOD1 remark-ably inhibits the proliferation , migration and invasion abilities of AIPC .

Objective To analyze the setup errors of external?beam and intracavitary radiotherapy for cervical cancer based on the data of cone?beam computed tomography ( CBCT) , and to provide a basis for clinical practice. Methods Sixty patients with stage ⅡA?ⅢB cervical cancer who were admitted to our hospital and underwent external?beam and intracavitary radiotherapy from March to June, 2015 were enrolled as subjects. Sixty patients with stage ⅡA?ⅢB cervical cancer undergoing conventional external?beam radiotherapy within the same period were also enrolled. The CBCT?based setup errors, containing setup errors in x, y, and z directions and three?dimensional vector deviation, in the initial treatment were obtained from each patient. Comparison of errors between the two treatment approaches was made by independent?samples t test. Results Fitting the data to a linear model revealed that the setup errors in x, y, and z directions increased with the increase in the distance between the position reference point and the center point of the target volume. External?beam radiation combined with intracavitary radiation had significantly reduced setup errors in x, y, and z directions than the conventional external?beam radiation ( 0.13± 0?12 vs. 0.31± 0?24, P=0?000;0.23±0?18 vs. 0.47±0?36, P=0?001;0.18±0?11 vs. 0.27±0?18, P=0?001). Conclusions In order to reduce the setup errors, CT scan needs not only a reference marker as close as possible to the center of the tumor, but also a reliable and accurate approach for postural fixation.

Background The incidence of dry eye is gradually increased,and researches showed that inflammation participated in the pathogenesis and development of dry eye.The current therapy for dry eye can only relieve symptom but not achieve final cure.Stem cell therapy has been used in the treatment of limbal stem cell deficiency.However,whether it is feasible for the stem cell treating dry eye is still unclear.Objective This study attempted to investigate a new approach to treat dry eye syndrom by using human umbilical cord mesenchymal stem cells (HUCMSCs).Methods The use and care of experimental animals complied with the Tsinghua University School of Medicine Laboratory Animal Care Details.HUCMSCs were cultured and cell suspension was prepared with the cell density of 5×105/ml.Twenty 24-week-old male NOD/Ltj mice were randomized into 4 groups.0.1 ml PBSHUCMSCs suspension was injected via tail vein or lacrimal respectively in the caudal vein injection group and lacrimal injection group,and 10 μ1 PBS-HUCMSCs suspension was topically administed in the eye drops group.The NOD/Ltj mice without any treatment served as the model group.Five male ICR mice were used as the normal control group.Tear secretion was quantitatively detected with phenol red cotton thread in 1,2,3 weeks after injection,and corneal epithelial defect was scored by fluorescein staining.The serum levels of interleukin (IL)-6,IL-17a,tumor necrosis factor-α (TNF-αt) and interferon-γ (IFN-γ) were assayed by ELISA.The relative expression levels of p65,Stat3,Stat5 and extracellular signal-regulated kinases (Erk)-1 in lacrimal gland were detected by Western blot.Results The tear secretion amount was significantly different among the model group,caudal vein injection group,lacrimal injection group and eye drops group in various time points (1 week:F =3.700,P =0.040;2 weeks:F =5.150,P =0.008;3 weeks:F=10.130,P＜0.001).The tear secretion amount was increased in the caudal vein injection group and lacrimal injection group compared with the model group in different time points (all at P＜0.05),and no significant difference was seen in tear secretion amount between eye drops group and model group among various time points (all at P＞0.05).The fluorescein staining score was 3.00±0.63,9.40±1.62,5.20±1.17,4.20±1.17 and 7.20±0.98 in the ICR mouse control group,model group,caudal vein injection group,lacrimal injection group and eye drops group 1 week after injection respectively,and the scores were significantly lower in the caudal vein injection group,lacrimal injection group and eye drops group than those in the model group (P =0.001,0.000,0.033).The serum levels of IL-6,IL-17a and TNF-α in the caudal vein injection group were evidently lower than those in the model group (t =4.70,3.46,11.0,all at P＜0.01),but no significant difference was displayed in the serum IFN-γ level among the five groups (F=1.740,P=0.170).The expressions of STAT5 were significantly decreased in the mice treated with tail vein injection and lacrimal injection compared with mice without treatment (both at P＜0.05).Conclusions Administration of HUCMSCs via intravenous and lacrimal injection can alleviate the inflammatory response during progression of dry eye syndrome by down-regulating the serum level and expression of inflammation-related factors in NOD/Ltj mice.The topical administration of HUCMSCs eye drops can attenuate the symptom of dry eyes by lubricating the cornea and suppling nutrition.

Objective No uniform standards has been established for the clinical diagnosis of schizophrenia .The article was to investigate the differentially expressed microRNA ( miRNA) profile and explore its clinical significance . Methods Differentially expressed miRNAs were screened by FlashTag TM Biotin RNA chips and SAM software in serum of patients with schizophrenia and healthy controls , then the validation was performed by real-time fluorescent quantitative reverse transcription polymerase chain reaction ( RT-PCR) . Results Three differentially expressed miRNAs were screened , including up-regulated hsa-miR-1281 ( fold change =1.50881) and two down-regulated miRNAs:hsa-miR-2861(fold change=0.642) and hsa-miR-638 (fold change=0.516).The com-parative analysis of RT-PCR by SPSS 17.0 Kruskal Wallis H Test validated the expression levels of hsa-miR-2861 and hsa-miR-638 in patients with schizophrenia decreased significantly in comparison to healthy controls (χ2 =4.089,χ2 =4.083, P<0.001).While the expression level of hsa-miR-1281 in patients with schizophrenia was in higher expression level compared with control group (χ2 =5.333, P<0.001). Conclusion There are differentially expressed miRNA profile in serum of patients with schizophrenia , in which miR-1281 , miR-2861 and miR-638 could be new serum markers for diagnosis of schizophrenia .