1.Optimization of honey processing technology of Phellinus igniarius by multi-index comprehensive score combined with analytic hierarchy process
Yidan HONG ; Wei WANG ; Liqiang ZHANG ; Yimei ZHANG ; Lihua LI ; Chengguo JU
China Pharmacy 2024;35(11):1309-1314
		                        		
		                        			
		                        			OBJECTIVE To optimize the honey processing technology of Phellinus igniarius. METHODS The key factors of honey processing technology of P. igniarius (honey-water ratio, the mass ratio of honey-water to P. igniarius, the frying temperature and the frying time) were investigated by orthogonal test combined with analytic hierarchy process to determine the optimal technological parameters, using the internal quality (the contents of ergosterol, protocatechuic aldehyde and protocatechuic acid) and appearance traits as evaluation indexes. RESULTS The optimal process of honey-roasting P. igniarius was to take raw P. igniarius (1 cm3 square block), add the appropriate amount of auxiliary materials (with 25 kg of refined honey and water for every 100 kg of P. igniarius), mix well, moisten for 2 h until the auxiliary materials were completely absorbed; put it in a frying container, fry at the frying temperature of 130-140 ℃ for 5 min; take it out, put it in an oven at 50 ℃ for 2 h; take it out, and let it cool. The RSD of the results of three validation experiments was 0.68%. CONCLUSIONS The optimized honey processing technology of P. igniarius is stable and feasible.
		                        		
		                        		
		                        		
		                        	
2.Optimization of honey processing technology of Phellinus igniarius by multi-index comprehensive score combined with analytic hierarchy process
Yidan HONG ; Wei WANG ; Liqiang ZHANG ; Yimei ZHANG ; Lihua LI ; Chengguo JU
China Pharmacy 2024;35(11):1309-1314
		                        		
		                        			
		                        			OBJECTIVE To optimize the honey processing technology of Phellinus igniarius. METHODS The key factors of honey processing technology of P. igniarius (honey-water ratio, the mass ratio of honey-water to P. igniarius, the frying temperature and the frying time) were investigated by orthogonal test combined with analytic hierarchy process to determine the optimal technological parameters, using the internal quality (the contents of ergosterol, protocatechuic aldehyde and protocatechuic acid) and appearance traits as evaluation indexes. RESULTS The optimal process of honey-roasting P. igniarius was to take raw P. igniarius (1 cm3 square block), add the appropriate amount of auxiliary materials (with 25 kg of refined honey and water for every 100 kg of P. igniarius), mix well, moisten for 2 h until the auxiliary materials were completely absorbed; put it in a frying container, fry at the frying temperature of 130-140 ℃ for 5 min; take it out, put it in an oven at 50 ℃ for 2 h; take it out, and let it cool. The RSD of the results of three validation experiments was 0.68%. CONCLUSIONS The optimized honey processing technology of P. igniarius is stable and feasible.
		                        		
		                        		
		                        		
		                        	
3.CEP33779 ameliorates pulmonary fibrosis associated with rheumatoid arthritis via the cross-talk between JAK2 and TGF-β 1 signaling pathway
Yimei WEI ; Dandan WANG ; Xinmeng WANG ; Jie ZHANG
Chinese Journal of Rheumatology 2023;27(1):10-16,C1-1-C1-2
		                        		
		                        			
		                        			Objective:Rat model of RA complicated with pulmonary fibrosis were constructed to observe the degree of improvement of pulmonary fibrosis in RA rats by JAK2 inhibitor CEP33779 and the possible mechanisms.Methods:①The RA models were constructed by subcutaneous injection of 0.2 ml (1 mg/ml) of bovine type Ⅱ collagen into the tail of the rats on the day of modeling development (d0); intratracheal injection of 100 μl bleomycin (2.5 mg/kg) was used to induce pulmonary fibrosis model at d13. In vivo study: model rats were randomly divided into the normal group, pulmonary fibrosis group, pulmonary fibrosis CEP treatment group, RA complicated with pulmonary fibrosis group, and RA complicated pulmonary fibrosis CEP treatment group. Rats in the treatment group was given CEP (10 ml/kg) qd by gavage from d14 to week 4. The right hind foot of the rats was measured for joints swelling and the arthritis index score were measured, lung compliance (Cst) and lung specific gravity were measured. In addition, the pathological changes of the left lung were observed by HE and Masson staining, and the extracellular matrix level of the right lung was measured by protein immunoblotting (WB). ② In vitro study: TGF-β 1 (10 ng/ml) was applied to stimulate human embryonic lung fibroblasts (HFL1) for 24 h and 48h, and p-JAK2 expression was detected by immunofluorescence. After HFL1 inoculation of culture plates, the control group, TGF-β 1 stimulation group, TGF-β 1+ LY2109761 (TGFβ-R1/2 inhibitor group, 0.5 mmol/L and 2 mmol/L) group, TGF-β 1+CEP (0.1 mmol/L and 1.0 mmol/L) group were co-incubated for 48 h, and the expression levels of TGFβ-R2, α-SMA, JAK2, and Col 1 were measured by WB. Comparisons between multiple groups were made by Tukey′s test, and comparisons between the two groups were analyzed by independent t-test. Results:① In vivo study, compared with the control group (1.45±0.04), joint swelling was increased at d13 [(2.54±0.16) in RA+PF+Vehicle group, t=16.02, P<0.001], and the mean arthritis index score and toe volume were decreased 3 days after CEP treatment(d16) [(2.89±0.11), t=5.78, P<0.001; (1.92±0.13), t=6.85, P<0.001]. For rats with pulmonary fibrosis, all had different degrees of lung enlargement, increased lung specific gravity, decreased Cst, and increased lung inflammation and fibrosis[(0.96±0.06), t=19.76, P<0.001; (0.26±0.09), t=17.64, P<0.001; (3.63±1.51), t=6.00, P<0.001; (1.75±0.71), t=5.84, P<0.001]. After CEP gavage, rats that had RA complicated with pulmonary fibrosis had reduced lung swelling, decreased lung specific gravity, increased Cst [(0.82±0.05), t=5.76, P<0.001; (0.43±0.18), t=2.31, P=0.038], and the scores of pulmonary fibrosis and inflammation [(3.00±1.00); (1.56±0.52)] all showed a trend of decrease, but did not reach statistical difference CEP inhibited the expression of TGF-β 1, TGFβ-R2, α-SMA, Fn and JAK2 in lung tissue of pulmonary fibrosis rats, and the differences among the five groups were statistically significant ( F=9.02, P=0.017; F=4.86, P=0.048; F=6.57, P=0.032; F=11.26, P=0.010; F=13.32, P=0.007). ② In vitro study, TGF-β 1 stimulated HFL1 showed stronger phosphorylated JAK2 (p-JAK2) fluorescent signal WB showed a significant increase in the expression of TGFβ-R2, α-SMA, JAK2 and Col1, and after LY and CEP intervention, the above proteins were reduced in a concentration-dependent manner, with statistically significant differences among all five groups ( F=337.30, P<0.001; F=20.61, P<0.001; F=100.60, P<0.001; F=180.90, P<0.001). Conclusion:JAK2 inhibitors can ameliorate RA-related pulmonary fibrosis, and the mechanism may be through interfering with the "crosstalk" between JAK2 and TGF-β 1 signaling pathway.
		                        		
		                        		
		                        		
		                        	
4.Construction of the evaluation index system for comprehensive budget management in public hospitals
Wei HONG ; Yimei SUN ; Ronghua DENG ; Yizhi ZHANG ; Weiyun CHEN
Chinese Journal of Hospital Administration 2023;39(9):656-661
		                        		
		                        			
		                        			Objective:To construct an evaluation index system for comprehensive budget management at public hospitals and conduct empirical research for reference in evaluation of comprehensive budget management at public hospitals.Methods:From May to June 2022, based on the Implementation Measures for the Comprehensive Budget Management System of Public Hospitals and its interpretations, literature analysis, and expert interviews, a preliminary evaluation index system was developed for comprehensive budget management of public hospitals. Then the Delphi method was applied for two rounds of expert consultation, with experts invited to evaluate the importance of indexes, while the analytic hierarchy process was used to calculate the weight values of indexes. Based on such a comprehensive budget management evaluation index system, a questionnaire was developed, and tertiary public hospitals were selected based on the principle of convenient sampling. Then a questionnaire survey was conducted from June to October 2022. The questionnaire was filled out by hospital level leaders(chief accountants), finance/audit leaders, and directors of finance/audit/operation management departments from the sample hospitals with professional knowledge and rich operational management experience in budget management. The comprehensive budget management level of the sample hospital was thus evaluated based on the survey results. Results:The comprehensive budget management evaluation index system for public hospitals was composed of 2 first-level indexes, 4 second-level indexes, and 20 third-level ones. The weights for the 2 first-level indexes of target building and management orientation were both 0.500, while the weights for the 4 second-level indexes, namely strategic implementation, comprehensive coverage, performance evaluation, and adaptation matching were all 0.250. Among the third-level indexes, the weights of budget execution, system improvement, system establishment, organizational construction, and revenue and expenditure business ranked top five, with weights of 0.070, 0.069, 0.066, 0.066, and 0.064, respectively. A total of 95 hospitals were selected for questionnaire survey, and the evaluation found 46 of them with a comprehensive budget management level of " excellent" and " good" , 27 with a " moderate" level, and 22 with a " poor" and " very poor" level. Scores of the third-level indexes were between 39-94 points, while the score of 6 third-level indexes were below 60 points, namely interconnection, budget analysis, compatibility and adaptation, system improvement, participation level, and assessment application.Conclusions:The evaluation index system for comprehensive budget management at public hospitals in this study proves rational and practical, serving as a reference for the evaluation of comprehensive budget management at public hospitals and related researches.
		                        		
		                        		
		                        		
		                        	
5.Longitudinal study on the trajectory and influencing factors of cancer-related fatigue in breast cancer patients during chemotherapy
Junwei MA ; Yimei ZHAO ; Shan YE ; Ping LU ; Shengmei QIN ; Yuqiao ZHUO ; Wei ZHENG ; Juan YANG ; Lei WANG ; Qi LI ; Zhaohui GENG
Chinese Journal of Practical Nursing 2022;38(15):1121-1129
		                        		
		                        			
		                        			Objective:To investigate the longitudinal trajectory and influencing factors of cancer-related fatigue (CRF) in breast cancer patients during chemotherapy.Methods:From March 2019 to January 2020, breast cancer patients in Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine and Peking Union Medical College Hospital were selected as the research objects to conduct follow-up investigation. The survey tools included general information questionnaire, Cancer-related Fatigue Assessment Scale, International Physical Activity Questionnaire, and the Pittsburgh Sleep Quality Index Scale.Results:A total of 91 patients were included in the study. The incidence of severe CRF at each time point before chemotherapy, 3 weeks after chemotherapy and 6 weeks after chemotherapy were 1.1% (1/91), 8.8% (5/57) and 2.1% (1/48), respectively. The results of the generalized estimation equation showed that the trajectory of the total score CRF firstly increased and then decreased, reaching a peak at 3 weeks after chemotherapy (35.45±13.07), and mild CRF change showed statistical difference ( P<0.05). In addition, BMI and sleep and daytime dysfunction were the influencing factors of the total score of CRF. Disease stage, sleep disturbance and daytime dysfunction were the influencing factors of CRF with different severity. Conclusions:CRF is a prominent problem in breast cancer patients during chemotherapy. Attention should be paid to high-risk individuals with abnormal BMI and daytime function by medical staff.
		                        		
		                        		
		                        		
		                        	
6.Nucleomodulin BspJ as an effector promotes the colonization of Brucella abortus in the host
Zhongchen MA ; Shuifa YU ; Kejian CHENG ; Yuhe MIAO ; Yimei XU ; Ruirui HU ; Wei ZHENG ; Jihai YI ; Huan ZHANG ; Ruirui LI ; Zhiqiang LI ; Yong WANG ; Chuangfu CHEN
Journal of Veterinary Science 2022;23(1):e8-
		                        		
		                        			 Background:
		                        			Brucella infection induces brucellosis, a zoonotic disease. The intracellular circulation process and virulence of Brucella mainly depend on its type IV secretion system (T4SS) expressing secretory effectors. Secreted protein BspJ is a nucleomodulin of Brucella that invades the host cell nucleus. BspJ mediates host energy synthesis and apoptosis through interaction with proteins. However, the mechanism of BspJ as it affects the intracellular survival of Brucella remains to be clarified. 
		                        		
		                        			Objectives:
		                        			To verify the functions of nucleomodulin BspJ in Brucella's intracellular infection cycles. 
		                        		
		                        			Methods:
		                        			Constructed Brucella abortus BspJ gene deletion strain (B. abortus ΔBspJ) and complement strain (B. abortus pBspJ) and studied their roles in the proliferation of Brucella both in vivo and in vitro. 
		                        		
		                        			Results:
		                        			BspJ gene deletion reduced the survival and intracellular proliferation of Brucellaat the replicating Brucella-containing vacuoles (rBCV) stage. Compared with the parent strain, the colonization ability of the bacteria in mice was significantly reduced, causing less inflammatory infiltration and pathological damage. We also found that the knockout of BspJ altered the secretion of cytokines (interleukin [IL]-6, IL-1β, IL-10, tumor necrosis factor-α, interferon-γ) in host cells and in mice to affect the intracellular survival of Brucella. 
		                        		
		                        			Conclusions
		                        			BspJ is extremely important for the circulatory proliferation of Brucella in the host, and it may be involved in a previously unknown mechanism of Brucella's intracellular survival. 
		                        		
		                        		
		                        		
		                        	
7.Value of peritoneal protein clearance as a predictor of cardiovascular outcomes in peritoneal dialysis patients
Wei NIU ; Xiaoxiao YANG ; Yiwei SHEN ; Dahua MA ; Yimei XU ; Qianhui SONG ; Zanzhe YU ; Hao YAN ; Zhenyuan LI ; Zhaohui NI ; Wei FANG
Chinese Journal of Nephrology 2021;37(7):576-582
		                        		
		                        			
		                        			Objective:To investigate the predictive value of peritoneal protein clearance (Pcl) for cardiovascular events and cardiovascular mortality in peritoneal dialysis (PD) patients.Methods:Eligible PD patients were prospectively enrolled from January 2014 to April 2015 in the PD Center of Renji Hospital, School of Medicine, Shanghai Jiao Tong University. All patients were followed up until death, withdrawing from PD, transferring to other centers, or the end of study period (October 1, 2018). The patients were divided into high Pcl group and low Pcl group by the median Pcl, and the differences of related indicators between the two groups were compared. A multiple linear regression model was used to analyze the influencing factors of Pcl. The Kaplan-Meier method and Log-rank test were used to compare the cumulative survival rates of patients between the two groups. A multivariate Cox regression model was used to estimate the risk of cardiovascular events and cardiovascular mortality in relation to Pcl in PD patients.Results:A total of 271 patients were enrolled, with 135 males (49.8%), age of (56.92±0.84) years old and a median PD duration of 38.77(19.00, 63.10) months. There were 70 patients (25.8%) comorbiding with diabetes and 81 patients (29.9%) with cardiovascular diseases (CVD). The median Pcl of this cohort was 67.93(52.31, 88.36) ml/d. Compared with the low Pcl group (Pcl<67.93 ml/d), the high Pcl group (Pcl≥67.93 ml/d) had older age, and greater proportion of CVD, body mass index (BMI), pulse pressure, brain natriuretic peptide, mass transfer area coefficient of creatinine (MTACcr), and lower serum albumin (all P<0.05). There was no significant difference in gender, dialysis duration, proportion of diabetes, proportion of angiotensin converting enzyme inhibitor and angiotensin receptor blocker, proportion of continuous ambulatory PD, high sensitivity C reactive protein, fluid removal including 24 h urine volume and 24 h ultrafiltration, and residual renal function between the two groups (all P>0.05). Multiple linear regression analysis showed that serum albumin ( β=-0.388, P<0.001), BMI ( β=0.189, P<0.001), and MTACcr ( β=0.247, P<0.001) were independently related to lg(Pcl). During the study period, 55 patients experienced one or more cardiovascular events and 39 patients had cardiovascular mortality. According to Kaplan-Meier analysis, cardiovascular mortality in the high Pcl group was higher than that of low Pcl group (Log-rank χ2=6.902, P=0.009). Multivariate Cox regression analysis showed that, high lg(Pcl) was an independent influencing factor of cardiovascular events in PD patients ( HR=7.654, 95% CI 1.676-34.945, P=0.009). Conclusions:Serum albumin, BMI and MTACcr are independently associated with Pcl, and Pcl is an independent predictor of cardiovascular events in PD patients.
		                        		
		                        		
		                        		
		                        	
8.Hair Growth Promoting Effects of 650 nm Red Light Stimulation on Human Hair Follicles and Study of Its Mechanisms via RNA Sequencing Transcriptome Analysis
Kai YANG ; Yulong TANG ; Yanyun MA ; Qingmei LIU ; Yan HUANG ; Yuting ZHANG ; Xiangguang SHI ; Li ZHANG ; Yue ZHANG ; Ji’an WANG ; Yifei ZHU ; Wei LIU ; Yimei TAN ; Jinran LIN ; Wenyu WU
Annals of Dermatology 2021;33(6):553-561
		                        		
		                        			 Background:
		                        			Androgenetic alopecia (AGA) leads to thinning of scalp hair and affects 60%~70% of the adult population worldwide. Developing more effective treatments and studying its mechanism are of great significance. Previous clinical studies have revealed that hair growth is stimulated by 650-nm red light. 
		                        		
		                        			Objective:
		                        			This study aimed to explore the effect and mechanism of 650-nm red light on the treatment of AGA by using ex vivo hair follicle culture. 
		                        		
		                        			Methods:
		                        			Human hair follicles were obtained from hair transplant patients with AGA. Hair follicles were cultured in Williams E medium and treated with or without 650-nm red light.Real-time RT-PCR and immunofluorescence staining were used to detect the expression level of genes and proteins in hair follicles, respectively. RNA-sequencing analysis was carried out to reveal the distinct gene signatures upon 650 nm treatment. 
		                        		
		                        			Results:
		                        			Low-level 650 nm red light promoted the proliferation of human hair follicles in the experimental cultured-tissue model. Consistently, 650 nm red light significantly delayed the transition of hair cycle from anagen to catagen in vitro. RNA-seq analysis and gene clustering for the differentially expressed genes suggests that leukocyte transendothelial migration, metabolism, adherens junction and other biological process maybe involved in stimulation of hair follicles by 650-nm red light treatment. 
		                        		
		                        			Conclusion
		                        			The effect of 650-nm red light on ex vivo hair follicles and the transcriptome set which implicates the role of red light in promoting hair growth and reversing of miniaturization process of AGA were identified. 
		                        		
		                        		
		                        		
		                        	
9.Influencing factors of carotid-femoral pulse wave velocity and its prognostic value in peritoneal dialysis patients
Yimei XU ; Hao YAN ; Zanzhe YU ; Zhenyuan LI ; Dahua MA ; Yiwei SHEN ; Xinyu SU ; Jiangzi YUAN ; Zhaohui NI ; Wei FANG
Chinese Journal of Nephrology 2021;37(4):305-312
		                        		
		                        			
		                        			Objective:To evaluate the influencing factors of carotid-femoral pulse wave velocity (CF-PWV) and its value to predict outcomes in peritoneal dialysis (PD) patients.Methods:Eligible patients undergoing PD in Renji Hospital of Shanghai Jiao Tong University between August 2016 and July 2018 were recruited and prospectively followed up until death, PD cessation, or to the end of the study. CF-PWV was measured by an arterial pulse wave velocity meter to assess arterial stiffness (July 31, 2020). Overhydration was measured by bioimpedance spectroscopy. The patients were divided into CF-PWV≤10 m/s group and CF-PWV>10 m/s group according to the measured value of CF-PWV. The influencing factors of elevated CF-PWV were analyzed by multivariate logistic regression. Survival curves were generated using the Kaplan-Meier method and multivariate Cox proportional hazards models were used to analyze the difference for all-cause mortality and cardiovascular disease (CVD) mortality between the two groups.Results:A total of 224 PD patients were enrolled, including 133 males (59.4%). The age was (55.2±13.4) years old, and median PD vintage was 22.3(6.5, 59.3) months. Among them, 47(21.0%) patients were comorbid with diabetes, and 37(16.5%) patients had CVD history. The median CF-PWV was 9.6(8.4, 11.4) m/s for the cohort, and 105(46.9%) participants had CF-PWV over 10 m/s. Compared with CF-PWV≤10 m/s group, CF-PWV>10 m/s group patients had older age, increased percentage of diabetes and CVD (all P<0.05). Multivariate logistic analysis showed that increased age ( OR=1.070, 95% CI 1.043-1.099, P<0.001), diabetes ( OR=3.693, 95% CI 1.646-8.287, P=0.002) and higher overhydration ( OR=1.238, 95% CI 1.034-1.483, P=0.020) were independent influencing factors for elevated CF-PWV in PD patients. After followed up for 37.4(25.6, 41.7) months, 24 patients died, including 19 cases of CVD-related deaths. Kaplan-Meier survival analysis showed that all-cause mortality and CVD mortality were significantly higher in the CF-PWV>10 m/s group than those in CF-PWV≤10 m/s group (Log-rank χ2=6.423, P=0.011; Log-rank χ2=6.243, P=0.012, respectively). Multivariate Cox proportional hazards models showed that increased age was an independent influencing factor for both all-cause mortality and CVD mortality ( HR=1.057, 95% CI 1.010-1.107, P=0.018; HR=1.062, 95% CI 1.009-1.118, P=0.022). Conclusions:Increased arterial stiffness is relatively common in PD patients. Higher CF-PWV in PD patients is associated with increased age, diabetes and higher overhydration, and it is probably a valuable predictor of outcome in PD patients.
		                        		
		                        		
		                        		
		                        	
10.Study on the Effects of Gypenosides on Gene Expression of Major Urinary Proteins in Hypercholesterolemia Model Mice
Yanping YANG ; Yimei DU ; Lin QIN ; Wei WANG ; Yanliu LU ; Anjing LU ; Yao ZENG ; Daopeng TAN ; Yuqi HE
China Pharmacy 2020;31(15):1809-1815
		                        		
		                        			
		                        			OBJECTIVE:To investigate the effects of gypenosides (GPs)on gene expression of major urinary proteins (Mups) in liver tissue of hypercholesterolemia model mice. METHODS :C57BL/6J mice were divided into control (ND)group,model (HFD)group and GPs therapy (GP)group according to body weight (BW),with 11 mice in each group. Except for ND group , other groups were given high-lipid diet to induce hypercholesterolemia model. From the 17th week of feeding ,ND group and HFD group were given constant volume of 0.1%CMC-Na solution intragastrically ;GP group were given GPs suspension (250 mg/kg) intragastrically,once a day ,for consecutive 22 weeks. BW ,the levels of blood glucose (BG)and blood lipid (TC,LDL-C)were detected in each group. Total RNA of liver tissue was extracted ,and reverse transcription library was constructed and RNA-seq sequencing was performed. The differentially expressed genes were screened by PCA ,volcano map and scatter plot. RT-qPCR was used for verification for differentially expressed genes. The correlation between the expression of differentially expressed genes and the above pharmacodynamic indexes was analyzed by bivariate analysis. RESULTS :Compared with ND group ,BW,the levels of BG,TC and LDL-C in HFD group were increased significantly (P<0.05). Compared with HFD group ,above indexes of GP group were decreased significantly except for BW (P<0.05). PCA showed that the data of ND group and HFD group distributed in different quadrants ,and the data distribution of GP group was between above two groups. mRNA of Mup4,Mup5,Mup11,Mup15 and Mup21 in liver tissue of mice were increased significantly after treated with high-fat diet (P<0.05). mRNA of Mup3,Mup4, Mup5,Mup8,Mup12 and Mup21 were decreased significantly after treated with GPs (P<0.05). In ND group vs. HFD group and HFD group vs. GP group ,mRNA of Mup4,Mup5 and Mup21 genes changed significantly and the trend was opposite. Results of RT-qPCR verification showed that compared with ND group ,relative mRNA expression of Mup4,Mup5 and Mup21 gene were increased significantly in HFD group (P<0.05). Correlation analysis revealed that mRNA expression of Mup5 was positively correlated with the levels of TC and BG (r=0.727 1,0.670 6,P<0.05),mRNA expression of Mup4 and Mup21 were positively correlated with the level of BG (r=0.737 8,0.721 5,P<0.05). CONCLUSIONS :GPs can regulate the expression of Mups genes in liver tissue of hypercholesterolemia model mice , and reduce glucose and lipid level through regulating the mRNA over-expression of Mup4,Mup5 and Mup21.
		                        		
		                        		
		                        		
		                        	
            
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