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 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 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 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 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.

Objective To investigate the efficacy of dexmedetomidine combined with target-controlled infusion (TCI) of propofol and remifentanil for fiberoptic bronchoscopy in the elderly patients.Methods Forty ASA Ⅱ or Ⅲ patients,aged 65-75 yr,with body mass index of 20-30 kg/m2,scheduled for elective fiberoptic bronchoscopy,were randomly divided into 2 groups (n =20 each):control group (group C) and dexmedetomidine group (group D).In group D,a loading dose of dexmedetomidine 0.5/μg/kg was injected at 10 min before induction of anesthesia,followed by infusion at 0.5 μg· kg-1 · h-1 until the end of fiberoptic bronchoscopy.While the equal volume of normal saline was given instead in group C.Anesthesia was induced with TCI of propofol and remifentanil.The target effect-site concentration (Ce) of propofol was 3 μg/ml.When the plasma concentration and Ce were balanced,TCI of remifentanil (target Ce 4 ng/ml) was started.The fiberoptic bronchoscope was placed after consciousness was lost and then the Ces of propofol and remifentanil were adjusted to 1-3 μg/ml and 2-4 ng/ml,respectively.MAP,HR and OAA/S score were recorded before induction (T0),immediately after induction (T1),when the tip of fiberoptic bronchoscope reached the glottis (T2) and carina (T3),at the end of bronchoscopy (T4)and 10 min after the end of bronchoscopy (T5).The consumption of propofol and remifentanil,duration of bron-choscopy,emergence time,adverse cardiovascular events and side effects such as hyoxemia,nausea and vomiting,regurgitation and aspiration were recorded.Results Compared with group C,OAA/S score at T5 and the consumption of propofol and remifentanil was reduced,and emergence time was shortened,and the incidence of hypotension and hyoxemia was decreased in group D (P ＜ 0.05).No patients developed side effects such as hyoxemia,nausea and vomiting,regurgitation and aspiration in both groups.Conclusion Dexmedetomidine (infusion at 0.5 μg·kg-1 ·h-1 after a loading dose of 0.5 μg/kg) combined with TCI of propofol and remifentanil can be safely and effectively used for fiberoptic bronchoscopy in the elderly patients.

BACKGROUND:The main component of cartilage, type Ⅱ col agen gene expression in chondrocyte is positively correlated with SOX9 concentration in a dose-dependent manner.
OBJECTIVE:To observe the variation of SOX9 and type Ⅱ col agen mRNA content at different periods in the differentiation process (osteogenic, chondrogenic, adipogenic induction) of mesenchymal stem cel s, and to explore the correlation of SOX9 expression and type Ⅱ col agen.
METHODS:Bone marrow mesenchymal stem cel s were isolated from 4-week-old Kunming mice, and cultured in vitro to passage 3. The cel phenotype was identified with flow cytometry. Cel s were divided into three groups and subjected to three kinds of induction conditions favorable for adipogenic, chondrogenic and osteogenic differentiation, and each group was observed at three time points. In addition, the non-induced cel s were used as a control group. The total RNA of cel s was extracted at 3, 7, 14 days after induction, and SOX9 and type Ⅱ col agen mRNA was quantified with reverse transcription-polymerase chain reaction. The induced cel s were stained by immunofluorescence to observe the differentiation and perform statistical analysis.
RESULTS AND CONCLUSION:Passage 3 bone marrow mesenchymal stem cel s grew wel , and cel phenotype was confirmed as stem cel s by flow cytometry. The staining results showed that, the cel s differentiated into chondrocytes, adipocytes and osteoblasts. The SOX9 mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in osteogenic differentiation group, and the lowest in adipogenic differentiation group. Type Ⅱ col agen mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in adipogenic differentiation group, and the lowest in osteogenic differentiation group. SOX9 expression in chondrogenic differentiation group increased at 3 and 7 days, and then decreased at 14 days. While type Ⅱ col agen expression increased at 3, 7, 14 days. SOX9 mRNA levels increased as the osteogenic differentiation, while type Ⅱ col agen expression gradual y decreased. There was no significant difference in the SOX9 mRNA expression between adipogenic differentiation group and control group (P>0.05), while type Ⅱ col agen expression was not regularly changed. Experimental findings suggest that, critical effect of SOX9 in chondrogenic differentiation is better than that in osteogenic and adipogenic differentiation. SOX9 is associated with type Ⅱcol agen, which may alter along with the SOX9 in the early chondrogenic differentiation;SOX9 may play a fine-tuning role in the process of chondrogenic and osteogenic differentiation.

Objective To observe effects of electro-acupuncture (EA) on the expression of apoptosis related protein B-cell lymphoma/leukemia gene 2 (Bcl-2) and Bcl-associated protein X (Bax) in the hippoeampal CA1 region of rats with cerebral infarction,and on their behavior. MethodsAn animal model of cerebral ischemia was established by right side middle cerebral artery occlusion using thread in 48 male,adult Wistar rats,who were then randomly divided equally into an EA group and a control group.Both groups were sub-divided into 1 week,2 weeks and 3 weeks subgroups.The EA group began receiving EA 24h after the occlusion,applied at the Baihui (DU20) and Dazhui (BU14) points,once daily,for one,two or three weeks.The control group was reared conventionally and was not given any special treatment.The rats'learning and memory,motion and neural function were evaluated.The expression of Bcl-2 and Bax in the CA1 region of the hippocampus on the infarcted side and changes of apoptosis indexes were detected using immunohistochemical techniques.ResultsThe learning,memory,motion and neural function of the EA group rats were better on average than those of the control group at all observation time points.The expression of Bcl-2 protein increased along with reduced expression of Bax protein in the EA group significantly more than among the controls.TUNEL positive cells in the CA1 region of the hippocampus were reduced significantly in the EA group compared with the control group.ConclusionsBehavioral ability after cerebral infarction can be improved by EA at the Baihui (DU20) and Dazhui (BU14) points.EA can up-regulate the expression of Bcl-2 and down-regulate the expression of Bax and reduce TUNEL positive cells in the CA1 region of the hippocampus after cerebral infarction,which might be the mechanism of its neuroprotection.

Duck circovirus(DuCV),a potential immunosuppressive virus,was investigated in Southern China from March 2006 to December 2009 by using a polymerase chain reaction(PCR)based method. In this study,a total of 138 sick or dead duck samples from 18 different farms were examined with an average DuCV infection rate of～35%. It was found that ducks between the ages of 40～60 days were more susceptible to DuCV. There was no evidence showing that the DuCV virus was capable of vertical transmission. Farms with positive PCR results exhibited no regularly apparent clinical abnormalities such as feathering disorders,growth retardation or lower-than-average weight. The complete genomes of 9. strains from Fujian Province and 1 from Zhejiang Province were sequenced and analyzed. The 10 DuCV genomes,compared with others genomes downloaded from GenBank,ranged in size from 1988 to 1996 base pairs,with sequence identities ranging from 83.2% to 99.8%. Phylogenetic analysis based on genome sequences demonstrated that DuCVs can be divided into two distinct genetic genotypes,Group I(the Euro-USA lineage)and Group II(the Taiwan lineage),with approximately 10.0% genetic difference between the two types. Molecular epidemiological data suggest there is no obvious difference among DuCV strains isolated from different geographic locations or different species,including Duck,Muscovy duck,Mule duck,Cheery duck,Mulard duck and Pekin duck.