Objective To construct the eukaryotic expression vector for mouse microRNA miR-21 and identification its expression activ-ity in 293 cells. Methods The genomic sequence containing pre-miR-21 was amplified from mouse genomic DNA by PCR and cloned into the pRC/CMV plasmid. The constructed recombinant plasmid pRC/CMV-mmu-miR-21 was transfected to 293 cells by lipofectamine 2000, and the stably transfected cells were screened with G418,from which total RNA was extracted for detecting the expression of mature miR-21 by northern blot. In the meantime,a luciferase report plasmid examing the activity of miR-21 named pmiR-21-Luc reporter was also construc-ted,and luciferase activity analysis indicated the product of pRC/CMV-mmu-miR-21 indeed had biological activity. Results Both restriction enzyme digestion analysis and sequencing proved the recombinant plasmids were constructed correctly. The miR-21 was highly expressed in the screened clones of 293 cells and it had good biological activity. Conclusion The eukaryotic expression plasmid of mouse miR-21 was successfully constructed,which laid the foundation of further investigation of the role of miR-21 during skin wound healing.

Objective To evaluate the importance of three-dimensional radiotherapy for elderly patients of stage Ⅳ non-small cell lung cancer (NSCLC).Methods Comparing with treatment outcome of ≥65 years 67 patients and ＜ 65 years 134 patients using concurrent chemotherapy and thoracic threedimensional radiotherapy during 2003 to 2010 years.Survival analysis was taken by Kaplan-Meier method.The multivariate prognosis was analyzed by Cox model.Results The follow-up was 97.8％.The percentage of ≥65 years and ＜ 65 years patients accepted with concurrent 4-5 cycles chemotherapy were 30％ and 55％,and with 42％ and 49％ patients with radiotherapy ≥63 Gy.The median survival time (MST) were 17 months and 14 months (x2 =0.76,P =0.384) for ≥65 years and ＜ 65 years patients accepted with concurrent 4-5 cycles chemotherapy concurrent ≥63 Gy radiotherapy respectively.The MST and 1-,2-,3year overall survival rate were 17 months and 8 months,65％ and 23％,30％ and 13％,24％ and 9％(x2 =7.90,P =0.005) for whole groups patients treated with chemotherapy concurrent ≥63 Gy and ＜ 63 Gy radiotherapy.And the MST of patients ≥ 63 Gy was significantly longer than those with ＜ 63 Gy either concurrent chemotherapy any cycles (x2 =9.54,P =0.023).The MST were 14 months and 8 months (x2 =1.82,P=0.178),17 months and 17 months (x2 =0.47,P=0.492) for ≥ 65 years and ≥ 63 Gy radiotherapy patients accepted with concurrent 4-5 cycles and 2-3 cycles chemotherapy concurrent respectively.Multivariate analysis showed local response (β =0.600,P =0.003) and numbers of tumor metastasis (β =0.670,P =0.040) were independent factors for survival.Conclusions For a part of elderly patients of stage Ⅳ NSCLC,concurrent chemotherapy and thoracic three-dimensional radiotherapy can prolong survival time with acceptable toxicity.Perhaps radiotherapy is more important.

Objective To explore high-risk factors that affect the severity of diabetic foot ulcers and the relationship between the severity of diabetic foot ulcers and the general somatic conditions by analyzing clinical data of 218 diabetic foot patients.Methods A retrospective analysis of the clinical data of 218 diabetic foot patients,admitted from 2006 to 2010 was performed.Fasting and postprandial blood glucose ( FBG and PBG ),glycosylated hemoglobin,hemoglobin,serum albumin,serum creatinine,as well as serum urea nitrogen and fibrinogen were determined in patients with different Wagner levels.The effects of peripheral arterial disease,neuropathy,infections and other factors on severity of diabetic foot ulcers,and the prognosis of those patients with different Wagner levels were analyzed.Results ( 1 ) FBG and PBG in Wagner 5 patients were significantly higher than those in Wagner 1,but there was no statistical significance in HbA1C difference.(2) Hemoglobin,total cholesterol,serum albumin in Wagner 5 patients were decreased significantly ; white blood cells,serum creatinine and fibrinogen were increased significantly ( P＜0.01 ).(3) The intima-media thickness(IMT) of femoral artery in Wagner 2-and-above patients was increased significantly compared with that in Wagner 1 ( P＜0.05 ).( 4 ) Logistic regression analysis showed that hemoglobin,fibrinogen,glycated hemoglobin,femoral artery intima-media thickness,and bacterial infection were correlated independently with the severity of diabetic foot ulcers.(5) The hospitalized days were increased with rising Wagner levels ; the prognosis of Wagner 5 patients was the worst,with the highest mortality in the group.Conclusions Changes in blood level of hemoglobin,fibrinogen,along with higher HbA1C and femoral artery IMT,as well as infections are high-risk factors of severity in diabetic foot patients.HbA1C is not a good indicator of blood glucose control in patients with Wagner 5 grade.The higher the Wagner grade,the longer the time spent in hospital,and the worse the prognosis wouldbe.

ObjectiveTo prospectively investigate the impact of short-time response on survival of concurrent chemotherapy and thoracic three-dimensional radiotherapy (CCTTRT) for stage Ⅳ non-small cell lung cancer (NSCLC). Methods From Jan.2003 to Oct.2010,201 patients with pathologically or cytologically proven stage Ⅳ NSCLC were included.All patients received platinum-based chemotherapy.Of the 167 patients eligible for analysis,the median number of chemotherapy were 4 cycles.The median dose for planning target volume (PTV) of thoracic primary tumor was 63 Gy.Response was scored according to WHO criteria. Survival was calculated by Kaplan-Meier method and compared using the Logrank. Cox regression model were used to examine the effect of response on overall survival.ResultsThe follow-up rate of 201 patients was 97.5％.with 201,170 and 134 patients finished ＜ 1,1 -2 and ≥3 years' follow-up.For the 167 patients eligible for analysis,the CR,PR,NC and PD rate of primary tumor was 5.4％,65.9％,21.0％ and 7.7％,respectively.The effective group ( CR + PR) and ineffective group ( NC + PD) was 71.3％ and 28.7％,respectively.The median survival time (MST) for patients with CR,PR,NC and PD was 22.6,13.4,8.8 and 4.8 months,respectively ( χ2 =44.79,P =0.000).The MST for effective and ineffective group was 13.9 and 7.6 months,respectively in the whole group ( χ2 =8.3 0,P =0.004 ),12.1months and 7.3 months in those treated with 2 - 3 cycles chemotherapy ( χ2 =7.71,P =0.007 ),and 13.9months and 7.9 months in those treated with 2 -5 cycles chemotherapy and radiation dose to PTV ≥36 Gy ( χ2 =4.00,P =0.045 ).No significant MST difference was detected between patients of effective group and ineffective group treated with 4 -5 cycles chemotherapy ( χ2 =0.67,P =0.413),or those treated with 4 -5 cycles of chemotherapy and radiation dose to primary lesion ≥36 Gy (χ2 =0.00,P =0.956).Multivariate analysis showed that 4-5 cycles of chemotherapy and CR and PR achieved in primary tumor (β =0.182,P=0.041 ) were independent favorable factors for survival. Conclusion CCTTRT can improve local control,and prolong the survival time for Stage Ⅳ NSCLC.

Objective To prospectively evaluate the survival of different metastasis organs with concurrent chemotherapy and thoracic three-dimensional radiotherapy (CCTTRT) for stage Ⅳ non-small cell lung cancer (NSCLC).Methods Two hundred and one patients of stage Ⅳ NSCLC were enrolled from January,2003 to July,2010.Of the 182 patients eligible for analysis,The number of patients with single-organ metastasis or multiple-organ metastasis was 107 and 75,respectively.Patients were treated by platinum-based chemotherapy,the median number of cycle was 4.The median dose to planning target volume of primary tumor (DTPTv) was 63 Gy.Survival was calculated by Kaplan-Meier method and compared using the Logrank.Results The follow-up rate of 201 patients was 97.5％.with 201,170 and 134 patients finished ＜ 1,1 -2 and ≥3 years'follow up.Of 182 patients,the 1-,2-,and 3-year overall survival (OS) rate and median survival time (MST) was 41.0％,17.0％,10.0％ and 10.5 months,respectively ;with single-organ metastasis and multi-organ metastasis were 50％,20％,14％ and 13 months and 29％,12％,0％ and 8.5 months ( x2 =10.10,P =0.001 ),respectively; compared with multi-organ metastasis,the 1-,2-,and 3-year OS arte and MST of patients with bone,lung metastasis only was 58％,25％,16％ and 14 months (x2 =10.42,P=0.001 ) and 49％,21％,21％ and 11 months (x2 =6.39,P=0.011 ) respectively;patients with brain metastasis only did not show advantage of survival comparing with patients with multi-organ metastasis (49％,8％,0％ and 12 months and 29％,12％,0％ and 8 months,respectively;x2 =0.71,P =0.401 ) ;the 1-,2-,and 3-year OS rate and MST was 63％,23％,19％ and 15 months and 42％,15％,0％ and 10 months,respectively for patients with single-organ metastasis and multi-organ metastasis patients who accepted 4 - 5 cycles of chemotherapy ( x2 =6.47,P =0.011 ) ; for patients under the same metastasis and 4 - 5 cycles of chemotherapy,no matter whether single-organ or multiple-organ metastases,the 1 -,2-,3-year OS rate and MST of patients with enough radiotherapy on DTPTV ≥63 Gy were better than patients without enough radiotherapy ( DTPTV ＜ 63 Gy ) ( 71％,25 ％,25％ and 16.8 months and 33％,17％,0％ and 10.5 months,respectively;x2 ＝4.73,P =0.030 ;54％,21％,0％ and 14.3 months and 29％,10％,0％ and 7.6 months,respectively,x2 =8.16,P =0.004).The MST of liver metastases was 6 months,there was significantly difference when comparing with non liver matastasis ( x2 =17.21,P =0.000).Conclusions It is very important to treat stage Ⅳ NSCLC with CCTTRT,especially patients with single-organ metastasis.Liver metastases is a unfavorable prognostic factor.

Objective To evaluate the overall survival and safety among patients for stage Ⅳ non-small cell lung cancer (NSCLC) treated with concurrent chemotherapy and thoracic three-dimensional radiotherapy (CCTTRT).Methods From Jan.2003 to July 2010,201 patients with stage Ⅳ NSCLC were included.All patients were treated with CCTTRT.Those patients who received only one cycle chemotherapy were not included in survival analysis,but analysis of toxicity.One hundred and eighty-two patients were eligible for survival analysis.All patients received platinum-based two-drug chemotherapy.The median number of cycles was 4.The median dose to planning target volume of primary tumor ( DTPTV ) was 63 Gy.Treatment-related gastrointestinal and hematological toxicity were scored according to WHO criteria.Radiation-related pneumonitis and esophagitis were evaluated according to the National Cancer Institute's Common Terminology Criteria for Adverse Events (CTC) version 3.0.Survival was calculated by Kaplan-Meier method and compared using the Logrank.Cox regression model was used to examine the effect of CCTTRT on overall survival.Results The follow-up rate of 201 patients was 97.5％.with 201,170 and 134 patients finished ＜ 1,1 -2 and ≥3 years' follow-up,respectively.Of the 182 patients eligible for survival analysis,further stratified analysis showed that the 1-,2-and 3-year overall survival rate and median survival time (MST) was 54％,20％,13％ and 14.3 months,respectively for patients treated with concurrent 4 -5 cycles chemotherapy and CCTTRT,and 66％,23％,19％ and 16.1 months,respectively for those treated with 4 -5 cycles chemotherapy and DTPTV ≥ 63 Gy.Under similar chemoradiotherapy intensity,the MST of patients with single organ metastasis was significantly longer than that with multiple organ metastases ( 13.0 months versus 8.5 months,x2 =10.10,P =0.001 ).For patients eligible for survival analysis and received 4 - 5 cycles of systemic chemotherapy,MST of patients treated with DTPTV≥63 Gy was significantly longer than those treated with DTPTV ＜63 Gy[14.9 months vs.8.4 months (x2 ＝20.48,P =0.000) and 16.1 months vs.8.8 months ( x2 =11.75,P =0.001 )].For patients with single organ metastasis,MST was 16 months for those treated with DTPTV ≥63 Gy and 9 months for those with DTPTV ＜63 Gy (x2 =10.51,P=0.000) ;for patients with multiple organ metastasis,it was 11 months and 7 months,respectively ( x2 =7.90,P =0.005 ).Multivariate analysis showed that concurrent 4 - 5 cycles chemotherapy and DTPTV ≥63 Gy (β =0.243,P=0.019) and improved KPS (β =1.268,P=0.000) were independent factors for survival.For the whole group,45％ patients had Grade 2 -3 gastrointestinal toxicity,35.0％ grade 3- 4 leukopenia,18％ grade 3- 4 thrombocytopenia.15.0％ grade 3- 4 anemia,9.5％ Grade 2 - 3 radiation pneumonia and 13.4％ radiation esophagitis,respectively.Conclusions For stage Ⅳ NSCLC,CCTTRT can prolong survival time with acceptable toxicity.Radiotherapy to thoracic primary tumor should be under consideration.

Objective To observe the therapeutic effect of Qilin Xintongshu pill for treatment of patients with non-ST segment elevation acute coronary syndromes(NSTE-ACS)accompanied by gastrointestinal bleeding (GIB). Methods A prospective randomized controlled trial was conducted. A total of 67 hospitalized patients in Cardiovascular Department of Qionghai City Hospital of Traditional Chinese Medicine were divided into therapy group(35 cases)and control group(32 cases). Both groups were given conventional medical treatment(except anti-coagulative agent). In the therapy group,the patients received Qilin Xintongshu pill(the ingredients consisting of panax pseudo-ginsen,dragon's blood,immature orange fruit,etc.)oral administration,once 5 mg and 3 times a day,while in the control group,the patients accepted clopidogrel orally,once 75 g and once a day,the therapeutic course in both groups being 30 days. The major cardiovascular events(such as death,a newly-happened myocardial infarction(MI),MI secondary,obstinately ischemia)and incidence of massive hemorrhage of gastrointestinal tract were compared between the two groups,and the adverse events were observed. Results After treatment,the incidence of cardiovascular events and massive hemorrhage of gastrointestinal tract in the therapy group were decreased significantly compared to those in the control group 〔incidence of cardiovascular events:8.57%(3/35)vs. 28.13%(9/32), incidence of massive hemorrhage of gastrointestinal tract:2.86%(1/35)vs. 21.88%(7/32),both P<0.05〕. Adverse events were not observed in both groups,and the examinations of blood,urine,liver and renal functions were of no abnormalities before and after treatment. Conclusion Qilin Xintongshu pill can effectively reduce the incidences of cardiovascular events and massive hemorrhage of gastrointestinal tract in patients with NSTE-ACS accompanied by GIB in the therapeutic course of 30 days,therefore this traditional Chinese herbal medicine is an ideal agent for treatment of such disease.

Objective To evaluate the clinical efficacy of Qilinxintongshu pill for treatment of critical patients with unstable angina pectoris (UAP) accompanied by upper alimentary tract damage.Methods A prospective randomized controlled trial was conducted. A total of 60 critical outpatients or inpatients with confirmed diagnosis of UAP accompanied by upper alimentary tract damage in Cardiology Department of Hospital of Traditional Chinese Medicine of Qionghai City from November 2011 to January 2015 were enrolled in the study, and they were assigned to a therapy group (30 cases) and a control group (30 cases) by table of random number. The conventional medical basic therapy was given to both groups, in addition, the patients in therapy group took Qilinxintongshu pill (made of notoginseng, dragon's blood, fruit of immature citron, etc.) 5 g once, three times a day, and the patients in control group orally administered clopidogrel 75 mg once a day, the therapeutic course being consecutive 30 days. After treatment, the rates of main cardiovascular events [including death, a newly-happened myocardial infarction (MI), and obstinate ischemia] and events of alimentary damage were compared between the two groups, and adverse effects were observed.Results There was no statistically significant difference in incidence of cardiovascular events between the therapy group and control group [6.7% (2/30) vs. 16.7% (5/30),P > 0.05]. The incidence of massive hemorrhage of gastrointestinal tract was significantly lower in therapy group than that in control group [10.0% (3/30) vs. 36.7% (11/30),P < 0.05]. In the treatment process, there was no significant untoward side effect, and no abnormalities in routine blood and urine tests, liver and renal functions were found.Conclusion Qilinxintongshu pill for treatment of critical patients with UAP accompanied by upper alimentary damage is safe and effective, and does not enhance the incidence of massive hemorrhage of gastrointestinal tract.

This study was aimed to establish a method for sensorial color digitalization of Chinese herbal medicines (CHMs) with the application of spectrocolorimeter. The discussion was focused on difficulties of distinguishing surface and section color of CHMs. Based on uniform color space system of CIE1976L*a*b*, two methods for determination of section and surface color were constructed with two different kinds of spectrocolorimeters taking Glycyrrhizae Radix et Rhizoma as the experimental objective. In this paper, different kinds of sample preparation methods were used. Based on results, the method of scraping and grinding was proposed to prepare samples for section color determination. The method of wet pressing and peeling was proposed to prepare samples for surface color determination. Besides, RSD and dE*ab were served as evaluation indexes. This paper provided a simple, rapid and reliable analysis method for the color determination of CHMs. It also gave insight to future research on digitalization and modernization of CHMs' organoleptic characteristics based on traditional macroscopic identification.

This study was aimed to apply the electronic nose (E-nose) in the research of traditional Chinese medicine (TCM). The discussion was made on difficulties of using E-nose. The solution plan was proposed and the discrimination model was established. It provided a simple, rapid and effective analysi method in the identification of TCM. It also provided new ideas for the research and application of gas sensor arrays. E-nose was used in the ex-traction of TCM scent characteristics. Based on ion mobility spectrometry of MOS sensor, the fingerprint of TCM scent was established. The maximum response value of the sensor was used as analysis index. According to the diffi-culties of identification, two solution plans were proposed. Firstly, different detectors were employed to complete the classification. Secondly, radial basis function (RBF) and random forests (RF) were combined and then a cascade classifier was constructed in order to achieve the maximum of information obtained in conditions where the number of measurements, metal oxide semiconductor sensors in E-nose was limited. The results showed that both plans were accurate and practical with relatively high upper correct judge rate and better cross-validation (The highest upper correct judge rates were 95% and 100%, 96% and 80%, respectively). It was concluded that this study firstly ap-plied cascade classifier in the establishment of TCM identification by E-nose. With limited amount of sensors, the maximum information was received through data mining. Using E-nose in the identification of TCM was rapid and accurate. The established pattern recognition method was maneuverable with accurate identification rate and stability compared to conventional sensory identification method. It provided a simple and rapid analysis method for the iden-tification of TCM.