1.Effect of Plasma Epstein-Barr Virus Nucleic Acid Loads on the Clinical Features and Prognosis in Adult Secondary Hemopha-gocytic Lymphohistiocytosis
Li-Min DUAN ; Guang-Li YIN ; Tian TIAN ; Ju-Juan WANG ; Xin GAO ; Wan-Ying CHENG ; Zi-Wei FANG ; Hong-Xia QIU ; Ji XU
Journal of Experimental Hematology 2024;32(4):1238-1247
		                        		
		                        			
		                        			Objective:To investigate the effect of pre-treatment plasma Epstein-Barr virus(EBV)DNA copy number on the clinical features and prognosis of patients with adult secondary hemophagocytic lymphohistiocytosis(sHLH).Methods:The clinical characteristics,survival rate,and prognostic factors of 171 patients with adult sHLH treated at Jiangsu Province Hospital from June 2017 to January 2022 were retrospectively analyzed in this study.Patients were divided into three groups,including the EBV DNA-negative group(<5.0 × 102 copies/ml),lower EBV-DNA loads group(5.0 × 102-8.51 × 104 copies/ml),and higher EBV-DNA loads group(>8.51 × 104 copies/ml),according to pre-treatment plasma EBV-DNA copy number.Cox regression model was established for screening prognostic factors.Adult sHLH survival prediction model was constructed and realized through the nomogram based on EBV-DNA load after adjusted the factors affecting survival of etiology and treatment strategy.Concordance index(C-index)and calibration curves were calculated to verify model predictive and discriminatory capacity.Results:Among 171 adult sHLH patients,84 patients were not infected with EBV(EBV DNA-negative group),and 87 with EBV(EBV DNA-positive group,48 lower EBV-DNA loads group and 39 higher EBV-DNA loads group).Consistent elevations in the levels of liver enzymes(ALT and AST),LDH,TG,β2-microglobulin and ferritin across the increasing of EBV-DNA load(all P<0.05),while the levels of fibrinogen decrease(P<0.001).The median follow-up time was 52 days(range 20-230 days),and 123 patients died.The overall survival(OS)rate of patients in EBV DNA-positive group was lower than that in EBV DNA-negative group(median OS:40 days vs 118 days,P<0.001).Higher EBV-DNA loads had worse OS(median OS:24 days vs 45 days vs 118 days,P<0.0001 for trend)compared to lower EBV-DNA loads and EBV DNA-negative group.Multivariate Cox analysis revealed that higher EBV-DNA loads(P=0.005),fibrinogen≤ 1.5 g/L(P=0.012),ferritin(P=0.041),associated lymphoma(P=0.002),and anti-tumor based strategy(P=0.001)were independent prognostic factors for OS.The C-indexes of 30 day,90 days,365 days survival rate were all greater than 0.8 of the nomogram model and calibration curves provided credibility to their predictive capability.Subgroup analysis showed that patients with higher EBV-DNA loads had a significantly worse prognosis in adult sHLH who were women,ferritin>5 000 μg/L,β2-microglobulin>7.4 mmol/L and regardless of age,etiologies,HScore points.Conclusion:The EBV-DNA load is a strong and independent predictor for survival in patients with sHLH.The prognostic nomogram based on EBV-DNA loads was dependable and provides a visual tool for evaluating the survival of adult sHLH.
		                        		
		                        		
		                        		
		                        	
2.Expression and Clinical Significance of Serum sFas/sFasL in Patients with Secondary Hemophagocytic Lymphohistiocytosis.
Ling Ling WANG ; Wan Ying CHEN ; Ju Juan WANG ; Guang Li YIN ; Li Min DUAN ; Tian TIAN ; Hong Xia QIU
Journal of Experimental Hematology 2023;31(3):889-895
		                        		
		                        			OBJECTIVE:
		                        			To investigate the expression and clinical significance of soluble Fas (sFas) and sFasL in patients with secondary hemophagocytic lymphohistiocytosis (sHLH).
		                        		
		                        			METHODS:
		                        			From September 2015 to December 2020, 86 sHLH patients who met the HLH2004 diagnostic criteria were collected. They were divided into 55 cases in the MAHLH group and 31 cases in the NonMAHLH group according to the etiology. Thirty healthy persons were chosen as the normal control group, and 20 patients with systemic lupus erythematosus (SLE) were chosen as the disease control group. The expression levels of sFas and sFasL in the serum of patients with each group were detected by ELISA, and the clinical data were collected for statistical analysis. The significance of sFas and sFasL in sHLH was analyzed by ROC curve.
		                        		
		                        			RESULTS:
		                        			Serum levels of sFas and sFasL in patients with newly diagnosed sHLH were significantly higher than those in disease control group and normal control group (P<0.01). The levels of sFas and sFasL in MAHLH group were significantly higher than those in nonMAHLH (infection related HLH and autoimmune disease related HLH) group (P<0.01). The serum levels of sFas and sFasL in 17 newly treated patients with sHLH (17/86) after treatment were significantly lower than those before treatment (P<0.01). The serum sFas level in newly diagnosed sHLH patients was positively correlated with SF(r=0.35), sCD25(r=0.79) and sFasL(r=0.73). The serum sFasL level was positively correlated with SF(r=0.39), sCD25(r=0.64) and sFas(r=0.73). Compared with the NonMAHLH group, the area under the ROC curve was 0.707 (95% CI: 0.593-0.821) (P=0.0015). The optimal critical value for diagnosing MAHLH by sFas level was 12 743 pg/ml, and the sensitivity and specificity were 70.9% and 71% respectively. Compared with the NonMAHLH group, the area under the ROC curve was 0.765(95% CI: 0.659-0.87)(P<0.01). The median OS time of sFas high expression group (≥16798.5 pg/ml) and sFasL high expression group (≥4 785 pg/ml) was significantly shorter than that of the low expression group (P<0.001).
		                        		
		                        			CONCLUSION
		                        			Serum levels of sFas and sFasL can be used for the early diagnosis and differential diagnosis of sHLH disease, and are the factor related to the poor prognosis of sHLH.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Lymphohistiocytosis, Hemophagocytic
		                        			;
		                        		
		                        			Clinical Relevance
		                        			;
		                        		
		                        			ROC Curve
		                        			;
		                        		
		                        			Sensitivity and Specificity
		                        			;
		                        		
		                        			Lupus Erythematosus, Systemic
		                        			
		                        		
		                        	
3.Evaluation of Renal Impairment in Patients with Diabetic Kidney Disease by Integrated Chinese and Western Medicine.
Yi-Lun QU ; Zhe-Yi DONG ; Hai-Mei CHENG ; Qian LIU ; Qian WANG ; Hong-Tao YANG ; Yong-Hui MAO ; Ji-Jun LI ; Hong-Fang LIU ; Yan-Qiu GENG ; Wen HUANG ; Wen-Hu LIU ; Hui-di XIE ; Fei PENG ; Shuang LI ; Shuang-Shuang JIANG ; Wei-Zhen LI ; Shu-Wei DUAN ; Zhe FENG ; Wei-Guang ZHANG ; Yu-Ning LIU ; Jin-Zhou TIAN ; Xiang-Mei CHEN
Chinese journal of integrative medicine 2023;29(4):308-315
		                        		
		                        			OBJECTIVE:
		                        			To investigate the factors related to renal impairment in patients with diabetic kidney disease (DKD) from the perspective of integrated Chinese and Western medicine.
		                        		
		                        			METHODS:
		                        			Totally 492 patients with DKD in 8 Chinese hospitals from October 2017 to July 2019 were included. According to Kidney Disease Improving Global Outcomes (KDIGO) staging guidelines, patients were divided into a chronic kidney disease (CKD) 1-3 group and a CKD 4-5 group. Clinical data were collected, and logistic regression was used to analyze the factors related to different CKD stages in DKD patients.
		                        		
		                        			RESULTS:
		                        			Demographically, male was a factor related to increased CKD staging in patients with DKD (OR=3.100, P=0.002). In clinical characteristics, course of diabetes >60 months (OR=3.562, P=0.010), anemia (OR=4.176, P<0.001), hyperuricemia (OR=3.352, P<0.001), massive albuminuria (OR=4.058, P=0.002), atherosclerosis (OR=2.153, P=0.007) and blood deficiency syndrome (OR=1.945, P=0.020) were factors related to increased CKD staging in patients with DKD.
		                        		
		                        			CONCLUSIONS
		                        			Male, course of diabetes >60 months, anemia, hyperuricemia, massive proteinuria, atherosclerosis, and blood deficiency syndrome might indicate more severe degree of renal function damage in patients with DKD. (Registration No. NCT03865914).
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Diabetes Mellitus, Type 2
		                        			;
		                        		
		                        			Diabetic Nephropathies
		                        			;
		                        		
		                        			Hyperuricemia
		                        			;
		                        		
		                        			Kidney
		                        			;
		                        		
		                        			Proteinuria
		                        			;
		                        		
		                        			Renal Insufficiency, Chronic/complications*
		                        			
		                        		
		                        	
4.Conceptual framework and its application of World Health Organization rehabilitation competency framework in rehabilitation human resource management
Fang WU ; Qi JI ; Zhouying QIU ; Hong SUN ; Anqiao LI ; Mingxue DUAN ; Qianqian GAO ; Weiqin CAI ; Wengui ZHENG
Chinese Journal of Rehabilitation Theory and Practice 2022;28(3):275-284
		                        		
		                        			
		                        			Objective To explore the application and method of World Health Organization Rehabilitation Competency Framework (RCF) in rehabilitation human resource management, and provide theoretical and practical reference for rehabilitation human resource management.Methods Based on the International Standard Classification of Occupations issued by the International Labour Organization, the application scope of RCF was systematically analyzed. The theoretical framework of rehabilitation human resource management based on RCF was constructed in combination with competency-based human resource management framework. On this basis, taking rehabilitation nurses as an example, the corresponding management methods and paths are proposed from four aspects of human resource management, including career management, recruitment and selection, training and development, and performance appraisal and salary management.Results Based on RCF, the theoretical framework of human resource management for rehabilitation nursing positions was constructed, and the main contents and processes of human resource management based on RCF were clarified from four aspects: recruitment and selection, training and development, performance appraisal and salary management, and career management. Occupational competency of rehabilitation professionals in International Standard Classification of Occupations was analyzed by RCF.Conclusion The rehabilitation competency framework constructed by WHO can be used to supplement the responsibilities of rehabilitation-related professionals in the International Standard Classification of Occupations issued by the International Labour Organization. RCF serves as a tool for practice managers and rehabilitation professionals in identifying selection criteria, learning objectives, and professional title assessment criteria. It can also be used in recruitment and selection, training and development, performance appraisal, salary management and career management in the field of rehabilitation human resource management.
		                        		
		                        		
		                        		
		                        	
5.Application of limiting antigen avidity enzyme immunoassay for estimating HIV-1 incidence in men who have sex with men.
Xi Jia TANG ; Lei Jing DUAN ; Wen Li LIANG ; Si CHENG ; Ting Li DONG ; Zhen XIE ; Kang Mai LIU ; Fei YU ; Zi Huang CHEN ; Guo Dong MI ; Liang LIANG ; Hong Jing YAN ; Lin CHEN ; Li LIN ; Dian Min KANG ; Xiao Bing FU ; Mao Feng QIU ; Zhen JIANG ; Jie XU ; Zun You WU
Chinese Journal of Epidemiology 2022;43(1):72-77
		                        		
		                        			
		                        			Objective: To estimate the incidence of HIV-1 infection in men who have sex with men (MSM) in key areas of China through HIV-1 limiting antigen avidity enzyme immunoassay (LAg-Avidity EIA), analyze the deviation from the actual results and identify influencing factors, and provided reference for improving the accuracy of estimation results. Methods: Based on the principle of the cohort randomized study design, 20 cities were selected in China based on population size and the number of HIV-positive MSM. The sample size was estimated to be 700 according to the HIV-1 infection rate in MSM. MSM mobile phone app. was used to establish a detection appointment and questionnaire system, and the baseline cross-sectional survey was conducted from April to November 2019. LAg-Avidity EIA was used to identify the recent infected samples. The incidence of HIV-1 infection was calculated and then adjusted based on the estimation formula designed by WHO. The influencing factors were identified by analyzing the sample collection and detection processes. Results: Among the 10 650 blood samples from the participants, 799 were HIV-positive in initial screening, in which 198 samples (24.78%) missed during confirmation test. Only 621 samples were received by the laboratory. After excluding misreported samples, 520 samples were qualified for testing. A total of 155 samples were eventually determined as recent infection through LAg-Avidity EIA; Based on the estimation formula , the incidence of HIV-1 infection in MSM in 20 cities was 4.06% (95%CI:3.27%-4.85%), it increased to 5.53% (95%CI: 4.45%-6.60%)after the adjusting for sample missing rate. When the sample missing rate and misreporting rate were both adjusted, the incidence of HIV-1 infection in the MSM increased to 5.66% (95%CI:4.67%-6.65%). The actual incidence of HIV-1 infection in MSM in the 20 cities might be between 4.06% and 5.66%. Conclusions: Sample missing and misreporting might cause the deviation of the estimation of HIV-1 infection incidence. It is important to ensure the sample source and the quality of sample collection and detection to reduce the deviation in the estimation of HIV-1 infection incidence.
		                        		
		                        		
		                        		
		                        			Cross-Sectional Studies
		                        			;
		                        		
		                        			HIV Infections/epidemiology*
		                        			;
		                        		
		                        			HIV-1
		                        			;
		                        		
		                        			Homosexuality, Male
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Immunoenzyme Techniques
		                        			;
		                        		
		                        			Incidence
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Sexual and Gender Minorities
		                        			
		                        		
		                        	
7.Human Serum-derived Extracellular Vesicles Protect A549 from PM
Qiu Lian ZHOU ; Yu Zheng BAI ; Juan GAO ; Yi DUAN ; Yi Cheng LYU ; Long Fei GUAN ; Kenneth ELKIN ; Yu Ling XIE ; Zheng JIAO ; Hong Yun WANG
Biomedical and Environmental Sciences 2021;34(1):40-49
		                        		
		                        			Objective:
		                        			Epidemiological studies reveal that exposure to fine particulate matter (aerodynamic diameter ≤ 2.5 μm, PM 
		                        		
		                        			Methods:
		                        			EVs were isolated from the serum of healthy subjects, quantified 
		                        		
		                        			Results:
		                        			PM 
		                        		
		                        			Conclusions
		                        			EVs treatment promotes cell survival and attenuates PM
		                        		
		                        		
		                        		
		                        			A549 Cells
		                        			;
		                        		
		                        			Air Pollutants/toxicity*
		                        			;
		                        		
		                        			Apoptosis/drug effects*
		                        			;
		                        		
		                        			Cell Survival/drug effects*
		                        			;
		                        		
		                        			Extracellular Vesicles
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Middle Aged
		                        			;
		                        		
		                        			Particulate Matter/toxicity*
		                        			;
		                        		
		                        			Protective Agents/pharmacology*
		                        			;
		                        		
		                        			Proto-Oncogene Proteins c-akt/metabolism*
		                        			;
		                        		
		                        			Serum
		                        			
		                        		
		                        	
8.Expression and Clinical Significance of Soluble B7-H3 in Patient with Secondary Hemophagocytic Lymphohistiocytosis.
Xin LYU ; Yue WANG ; Ling-Ling WANG ; Guang-Li YIN ; Ling-Ling LIU ; Li-Min DUAN ; Tian TIAN ; Ji XU ; Hong-Xia QIU
Journal of Experimental Hematology 2020;28(3):1006-1011
		                        		
		                        			OBJECTIVE:
		                        			To investigate the expression and clinical significance of soluble B7-H3 (sB7-H3) in patients with secondary hemophagocytic lymphohistiocytosis (sHLH).
		                        		
		                        			METHODS:
		                        			The plasma samples of 85 newly diagnosed sHLH patients from December 2012 to April 2018 were collected. The patients were divided into lymphoma-related HLH(LHLH)group and infection-related HLH(IHLH)group. The expression of sB7-H3 in plasma was detected by ELISA, and the clinical data were collected for analysis. Fifteen healthy people were chosen as control group.
		                        		
		                        			RESULTS:
		                        			The expression level of sB7-H3 in lymphoma-related HLH and infection-related HLH group significant increased as compared with the control group, (P<0.05), and the expression level of sB7-H3 in lymphoma-related HLH group was significant higher than that in infection-related HLH group [(35.75± 9.90) vs (28.70±8.95) ng/ml)] (P<0.001). There were no significant statistical difference in the expression of some clinical factors (including age, fever, splenomegaly, ANC, Plt, FIB, calcium ion, serum albumin, LDH, serum ferritin, sCD25) in lymphoma-related HLH and infection-related HLH group (P>0.05). The evaluation of expression and significance of sB7-H3 in sHLH by using ROC curve, showed that the area under ROC curve comparison of patients in lymphoma-related HLH group and infection-related HLH group was 0.718 (95% CI 0.610-0.810) (P=0.0002), and predicting the sensitivity and specificity of the lymphoma-related HLH patients were 77.36% and 59.38%, respectively. The best cut-off value of patients in sB7-H3 was 29.81 ng/ml, the overall survival time of sB7-H3 high-expression group (≥29.81 ng/ml) was significant shorter than that in low-expression group (<29.81 ng/ml) (24 vs 440 d) (P<0.001). The clinical factors affecting the survival status of sHLH patients were neutrophils, albumin, serum ferritin, serum calcium ions and sB7-H3 levels.
		                        		
		                        			CONCLUSION
		                        			sB7-H3 associates with poor prognosis of sHLH patients, and may be involved in disease progression.
		                        		
		                        		
		                        		
		                        			Enzyme-Linked Immunosorbent Assay
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Lymphohistiocytosis, Hemophagocytic
		                        			;
		                        		
		                        			Lymphoma
		                        			;
		                        		
		                        			ROC Curve
		                        			
		                        		
		                        	
9.Spectrometric analyses of larotaxel and larotaxel liposomes quantification by high performance liquid chromatography.
Xue Qi LI ; Jian Wei LI ; Qiu Hong LI ; Yan YAN ; Jia Lun DUAN ; Yi Nuo CUI ; Zhan Bo SU ; Qian LUO ; Jia Rui XU ; Ya Fei DU ; Gui Ling WANG ; Ying XIE ; Wan Liang LU
Journal of Peking University(Health Sciences) 2019;51(3):467-476
		                        		
		                        			OBJECTIVE:
		                        			Larotaxel is a new chemical structure drug, which has not been marketed worldwide. Accordingly, the standard identification and quantification methods for larotaxel remain unclear. The spectrometric analyses were performed for verifying weight molecular formula, molecular weight and chemical structure of larotaxel. Besides, a quantification method was developed for measuring larotaxel in the liposomes.
		                        		
		                        			METHODS:
		                        			The molecular formula, molecular weight and chemical structure of larotaxel were studied by using mass spectrometry (MS), infra-red (IR), nuclear magnetic resonance (NMR) and ultraviolet-visible (UV-vis) spectrometric techniques. The absorption wavelength of larotaxel was investigated by UV-vis spectrophotometry full-wavelength scanning. Besides, a quantification method was developed by high performance liquid chromatography (HPLC), and then validated by measuring the encapsulation efficacy of larotaxel liposomes.
		                        		
		                        			RESULTS:
		                        			The four spectral characteristics of larotaxel were revealed and the corresponding standard spectra were defined. It was confirmed that larotaxel had the structure of tricyclic diterpenoids, with the molecular formula of C45H53NO14, the molecular weight of 831.900 1, and the maximum absorption wavelength of 230 nm. The quantitative method of larotaxel was established by using HPLC with a reversed phase C18 column (5 μm, 250 mm×4.6 mm), a mobile phase of acetonitrile-water (75:25, volume/volume), and a detection wavelength of 230 nm. The validation study exhibited that the established HPLC method was stable, and had a high recovery and precision in the quantitative measurement of larotaxel in liposomes. In addition, a new kind of larotaxel liposomes was also successfully prepared. The particle size of the liposomes was about 105 nm, with an even size distribution. And the encapsulation efficiency of larotaxel in the liposomes was above 80%.
		                        		
		                        			CONCLUSION
		                        			The present study offers reference standard spectra of larotaxel, including MS, IR, NMR, and UV-vis, and confirms the molecular formula, molecular weight and chemical structure of larotaxel. Besides, the study develops a rapid HPLC method for quality control of larotaxel liposomes.
		                        		
		                        		
		                        		
		                        			Chromatography, High Pressure Liquid
		                        			;
		                        		
		                        			Liposomes
		                        			;
		                        		
		                        			Magnetic Resonance Spectroscopy
		                        			;
		                        		
		                        			Taxoids
		                        			
		                        		
		                        	
10.Characteristic dysbiosis of gut microbiota of Chinese patients with diarrhea-predominant irritable bowel syndrome by an insight into the pan-microbiome.
Zhe WANG ; Cong-Min XU ; Yi-Xuan LIU ; Xiao-Qi WANG ; Lu ZHANG ; Mo LI ; Shi-Wei ZHU ; Zhong-Jie XIE ; Pei-Hong WANG ; Li-Ping DUAN ; Huai-Qiu ZHU
Chinese Medical Journal 2019;132(8):889-904
		                        		
		                        			BACKGROUND:
		                        			Irritable bowel syndrome (IBS) is reported associated with the alteration of gut microbial composition termed as dysbiosis. However, the pathogenic mechanism of IBS remains unclear, while the studies of Chinese individuals are scarce. This study aimed to understand the concept of dysbiosis among patients with Chinese diarrhea-predominant IBS (IBS-D), as a degree of variance between the gut microbiomes of IBS-D population and that of a healthy population.
		                        		
		                        			METHODS:
		                        			The patients with IBS-D were recruited (assessed according to the Rome III criteria, by IBS symptom severity score) from the Outpatient Department of Gastroenterology of Peking University Third Hospital, and volunteers as healthy controls (HCs) were enrolled, during 2013. The 16S rRNA sequences were extracted from fecal samples. Ribosomal database project resources, basic local alignment search tool, and SparCC software were used to obtain the phylotype composition of samples and the internal interactions of the microbial community. Herein, the non-parametric test, Wilcoxon rank-sum test was carried out to find the statistical significance between HC and IBS-D groups. All the P values were adjusted to q values to decrease the error rate.
		                        		
		                        			RESULTS:
		                        			The study characterized the gut microbiomes of Chinese patients with IBS-D, and demonstrated that the dysbiosis could be characterized as directed alteration of the microbiome composition leading to greater disparity between relative abundance of two phyla, Bacteroidetes (Z = 4.77, q = 1.59 × 10) and Firmicutes (Z = -3.87, q = 5.83 × 10). Moreover, it indicated that the IBS symptom features were associated with the dysbiosis of whole gut microbiome, instead of one or several certain genera even they were dominating. Two genera, Bacteroides and Lachnospiracea incertae sedis, were identified as the core genera, meanwhile, the non-core genera contribute to a larger pan-microbiome of the gut microbiome. Furthermore, the dysbiosis in patients with IBS-D was associated with a reduction of network complexity of the interacted microbial community (HC vs. IBS-D: 639 vs. 154). The disordered metabolic functions of patients with IBS-D were identified as the potential influence of gut microbiome on the host (significant difference with q < 0.01 between HC and IBS-D).
		                        		
		                        			CONCLUSIONS
		                        			This study supported the view of the potential influence of gut microbiome on the symptom of Chinese patients with IBS-D, and further characterized dysbiosis in Chinese patients with IBS-D, thus provided more pathological evidences for IBS-D with the further understanding of dysbiosis.
		                        		
		                        		
		                        		
		                        			Diarrhea
		                        			;
		                        		
		                        			microbiology
		                        			;
		                        		
		                        			Dysbiosis
		                        			;
		                        		
		                        			microbiology
		                        			;
		                        		
		                        			Feces
		                        			;
		                        		
		                        			microbiology
		                        			;
		                        		
		                        			Gastrointestinal Microbiome
		                        			;
		                        		
		                        			genetics
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Irritable Bowel Syndrome
		                        			;
		                        		
		                        			microbiology
		                        			;
		                        		
		                        			Models, Theoretical
		                        			;
		                        		
		                        			RNA, Ribosomal, 16S
		                        			;
		                        		
		                        			genetics
		                        			
		                        		
		                        	
            

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