1.Effect of different stretching lengths of lingual movable wing on the adduction of mandibular anterior teeth:a biomechanical study
Guorui ZHANG ; Kunwu ZHANG ; Wenyuanfeng CHEN ; Yining LIU ; Duhong LI ; Xinzhu ZHANG ; Baocheng CAO
Chinese Journal of Tissue Engineering Research 2024;28(2):247-251
		                        		
		                        			
		                        			BACKGROUND:Lingual movable wing is a new type of lingual orthodontic technique and the different stretching lengths of the wring affect the torque control effect of anterior teeth.However,there is yet no related biomechanical research. OBJECTIVE:To investigate the displacement trend of dentition during adduction of mandibular anterior teeth and the effect of different wing stretching lengths on the biomechanical effect of mandibular anterior teeth. METHODS:The data of the mandible and lower dentition were collected by cone-beam CT and reconstructed using Mimics software to establish a three-dimensional finite element model of mandibular anterior teeth adducted by the lingual movable wing.The ANSYS software was used to analyze the initial displacement of the mandibular anterior teeth under the following conditions:A,2 mm stretching length;B,2.5 mm stretching length;C,3 mm stretching length;and D,3.5 mm stretching length. RESULTS AND CONCLUSION:The trend of initial displacement of lower dentition:The central incisors moved lingually with depression,the lateral incisors and canines moved mildly lingually with mesial lingual torsion,the second premolar was tilted distally with a marked lingual inclination and the first molar showed an overall mesial inclination with mesial crown eversion.Therefore,in the adduction cases of mandibular tooth extraction,attention should be paid to the lingual movement of the second premolar,which could be offset by corresponding techniques in clinic.The trend of anterior tooth displacement in all directions:from condition A to condition D,in the sagittal direction,the difference value in crown-root displacement of central incisors changed from-11.891 μm to-5.757 4 μm,indicating that the central incisor changes from oblique movement to overall movement.The difference value in crown-root displacement of lateral incisors changed from-11.828 1 μm to-6.711 45 μm,and that of canines changed from-7.572 3 μm to-4.695 5 μm,indicating that the oblique movement of the lateral incisors and canines is also changing to an overall movement.In the vertical direction,from condition A to condition D,the reduction of incisors was gradually increased,while that of canines was gradually decreased.These findings indicate that the stretching length of the wing can affect the oblique movement trend of the anterior teeth.As the wing continues to stretch,the torque control of the lower anterior teeth will become better.
		                        		
		                        		
		                        		
		                        	
2.Optical coherence tomography imaging features of Coats disease and their correlation with macular fibrosis
Ziyi ZHOU ; Guorui DOU ; Hongxiang YAN ; Guoheng ZHANG ; Jinting ZHU ; Dongjie SUN ; Zifeng ZHANG ; Manhong LI ; Yusheng WANG
Chinese Journal of Experimental Ophthalmology 2024;42(5):436-441
		                        		
		                        			
		                        			Objective:To analyze the optical coherence tomography (OCT) imaging characteristics in patients with Coats disease and their value in predicting macular fibrosis.Methods:A nested case-control study was performed.A total of 43 patients (43 eyes) diagnosed with Coats disease through color fundus photography, ocular B-scan ultrasonography, fundus fluorescein angiography, and spectral-domain OCT examination were enrolled from January 2008 to October 2021 at the Xijing Hospital.Among them, there were 40 males and 3 females, aged from 2 to 60 years old, with a median age of 13 years.Macular fibrosis was used as an indicator of poor prognosis, and patients were divided into two groups based on whether macular fibrosis occurred at the end of follow-up.The differences in OCT characteristics between two groups were compared and logistic regression analysis was used to identify the risk factors for macular fibrosis.This study adhered to the Declaration of Helsinki and was approved by the Ethics Committee of Xijing Hospital of Fourth Military Medical University (No.KY20202009-C-1).Results:The OCT clinical features of 43 cases of Coats disease included intraretinal hard exudates in 43 eyes (100%), subretinal fluid in 21 eyes (48.8%), macular cysts in 17 eyes (27.9%), subretinal exudates in 9 eyes (20.9%), anterior retinal hyperreflective dots in 7 eyes (16.3%), epiretinal membrane in 21 eyes (48.8%), and intraretinal fluid in 22 eyes (51.2%).In color fundus photos of 41 eyes, 38 eyes (93.0%) had hard exudates distributed in the posterior pole and 27 eyes (65.9%) had the mid-peripheral region.OCT examination showed that hard exudates were distributed in the inner nuclear layer in 35 eyes (81.4%) and the outer nuclear layer in 33 eyes (76.7%).Among 21 eyes with exudative retinal detachment detected by OCT, 9 eyes (42.9%) were detected by fundus photography and 18 eyes (85.7%) were detected by B-scan ultrasonography.The proportions of eyes with subretinal fluid and subretinal exudates were higher in the macular fibrosis group than in the non-macular fibrosis group, and the differences were statistically significant ( χ2=20.755, P<0.001; χ2=6.133, P=0.013).Logistic regression analysis showed that the presence of subretinal fluid was a risk factor for macular fibrosis (odds ratio=48.345, 95% confidence interval: 4.272-547.066, P=0.002). Conclusions:OCT examination can detect subretinal fluid, subretinal exudates, macular cysts, macular exudates, and hyperreflective spots in the retina of patients with Coats disease.Subretinal fluid is a risk factor for macular fibrosis.
		                        		
		                        		
		                        		
		                        	
3.Effect and mechanism of melatonin on streptozotocin-induced diabetic pulmonary fibrosis
Tinglyu FU ; Guorui LI ; Rui XIONG ; Ruyuan HE ; Bohao LIU ; Ning LI ; Qing GENG
Chinese Journal of Endocrinology and Metabolism 2024;40(6):505-514
		                        		
		                        			
		                        			Objective:To explore the effects of melatonin on streptozotocin(STZ)-induced diabetic pulmonary fibrosis and regulatory mechanisms.Methods:C57BL/6 mice were randomly divided into the control group, STZ group, STZ+ low-dose melatonin(5 mg/kg) group, STZ+ high-dose melatonin(30 mg/kg) group using random number table, and a single intraperitoneal injection of STZ(150 mg/kg) was administered to establish a diabetic pulmonary fibrosis mouse model. Two weeks later, blood glucose levels ≥16.7 mmol/L confirmed successful modeling. Subsequently, melatonin was administered orally for 4 weeks, and the mice were sacrificed at 16 weeks for tissue sampling. Human umbilical vein endothelial cells were divided into the control group(glucose concentration is 5.5 mmol/L), high glucose group(glucose concentration is 33.3 mmol/L), high glucose+ low-dose melatonin(5 μmol/L) group, high glucose+ high-dose melatonin(20 μmol/L) group, and cells in each group were collected for subsequent detection after drug stimulation. Masson staining and immunofluorescence staining were used to observe fibrotic lesions, Western blotting was used to detect the expression related proteins, and sirtuin 3(Sirt3) siRNA was transfected to knock down Sirt3.Results:Significant fibrotic lesions were observed in the lung tissue of the STZ group compared to the control group, however, the STZ+ low-dose melatonin group and STZ+ high-dose melatonin group showed reduced fibrosis compared to the STZ group. In addition, compared to the control group, the endothelial cell marker platelet endothelial cell adhesion molecule-1(CD31) was significantly decreased in the STZ/high glucose group( P<0.001; P<0.001), and the interstitial fibrosis markers collagen 3, Vimentin, and α-smooth muscle actin(α-SMA) were significantly increased( P<0.001, P=0.035, P<0.001; P<0.001, P<0.001, P<0.001), but these trends were partially reversed after melatonin treatment in the STZ/high glucose+ low-dose melatonin group and the STZ/high glucose+ high-dose melatonin group. Moreover, the protein expression of Sirt3 was significantly reduced in the STZ/high glucose group compared to the control group( P<0.001; P<0.001), while it was increased in the STZ/high glucose+ low-dose melatonin and STZ/high glucose+ high-dose melatonin groups compared to the STZ/high glucose group( P=0.047, P<0.001; P=0.048, P<0.001). After transfecting Sirt3 siRNA to knock down the expression of Sirt3, the endothelial cell marker CD31 was significantly reduced( P=0.026), and interstitial fibrosis markers collagen 3, Vimentin, and α-SMA were significantly increased in the high glucose+ high-dose melatonin+ Sirt3 siRNA group compared to the high glucose+ high-dose melatonin group( P<0.001, P<0.001, P<0.001). Conclusion:Melatonin inhibits endothelial-mesenchymal transition by activating Sirt3 expression, thereby alleviating pulmonary fibrosis in STZ-induced diabetic mice.
		                        		
		                        		
		                        		
		                        	
4.Investigation of target resistance kdr, ace-1 mutations in Anopheles gambiae s.l. in Togo
LIANG Guorui ; ZHOU Xian ; LI Guoming
China Tropical Medicine 2024;24(6):652-
		                        		
		                        			
		                        			Abstract:  Objective To investigate the allelic mutations of kdr and ace-1 in Anopheles gambiae s.l., providing a scientific basis for mosquito-borne disease prevention and control in the region. Methods    Mosquitoes were collected from six villages in two malaria-prone areas, Est-mono (Ogou koulidé, Kepssi, Igbomedji) and Anié (Kolocopé, Bana, Afolé). All mosquitoes were used for DNA extraction, species identification, and genotyping of kdr and ace-1. Results    An. coluzzii and An. gambiae, two closely related species within the An. gambiae complex, were identified across six collection sites, with An. gambiae being predominant, constituting approximately 96.53% of the samples. Among all mosquito samples detected, high-frequency mutations were observed at the kdr 1014 locus in the An. gambiae complex, with mutation frequencies reaching 100.0% in Ogou koulidé, Bana, and Afolé, 97.5% in Kepssi and Igbomedji, and 91.1% in Kolocopé district. The allelic frequency of 1014F ranged from 85.5% to 100%. Mutation frequency of An. coluzzii 1014 locus in all three regions was 100%, with an allelic frequency of 1014F ranging from 90% to 100%, indicating a high rate of homozygosity. The kdr 1575 mutation frequency in An. gambiae ranged from 6.6% to 24.3%, with the allelic frequency of 1575Y ranging from 3.2% to 12.2%, all detected mutations were heterozygous, and no mutations were detected at the 1575 locus in An. coluzzii. Mutations at the ace-1 G119 locus were also observed in the An. gambiae complex, with mutation frequencies in An. gambiae ranging from 8.8% to 15% and the allelic frequency of 119S from 4.4% to 7.5%. In An. coluzzii, mutation frequencies ranged from 0 to 20.0%, with an allelic frequency of 119S from 0 to 10%, and all detected 119S mutations were heterozygous. Conclusions    The high frequencies of the Anopheles gambiae kdr mutation observed suggest that malaria mosquito vectors in Togo may have developed a high level of resistance to pyrethroid insecticides and possibly a moderate resistance to organophosphorus or carbamate insecticides. This indicates that local vector management authorities should prioritize insecticide resistance issues in future malaria vector control and resistance management planning, and conduct regular monitoring.
		                        		
		                        		
		                        		
		                        	
5.Characterization of the antigens in inactivated porcine circovirus type 2 vaccines and virus-like particle vaccines by high-performance size-exclusion chromatography coupled with multi-angle laser light scattering.
Yuan XU ; Yanli YANG ; Xingqi ZOU ; Cui LI ; Yuanyuan ZHU ; Yixian QIN ; Yan LI ; Ya Nan SHENG ; Yebing LIU ; Guorui PENG ; Xiaoai XU ; Songping ZHANG ; Qizu ZHAO
Chinese Journal of Biotechnology 2022;38(8):2948-2958
		                        		
		                        			
		                        			This paper aims to detect the antigens in porcine circovirus type 2 (PCV2) vaccines by high-performance size-exclusion chromatography (HPSEC) coupled with multi-angle laser light scattering (MALLS). With purified inactivated PCV2 and PCV2 virus-like particles (VLP) as references, two inactivated vaccines (a and b) and two VLP vaccines (c and d) for PCV2 from four manufacturers were analyzed by HPSEC-MALLS after demulsification. The antigen peaks in HPSEC-MALLS were identified by PCV2 antigen test strips, Western blotting and transmission electron microscope (TEM). The repeatability and linearity of the method were investigated. The results showed the virus antigens in the two inactivated vaccines were eluted at about 13.3 min in HPSEC. The molecular weight of these antigens was 2.61×106 (±4.34%) Da and 2.40×106 (±2.51%) Da, respectively, as calculated by MALLS. The antigen peaks of the two VLP vaccines also appeared at 13.3 min and the molecular weight was 2.09×106 (±2.94%) Da and 2.88×106 (±11.85%) Da, respectively, which was close to the theoretical molecular weight of PCV2. Moreover, an antigen peak of VLP vaccine c was observed at 11.4 min and the molecular weight was 4.37×106 (±0.42%) Da. The antigen was verified to be the dimer of VLP by TEM. Vaccine d and purified Cap VLP antigens were tested repeatedly, and the RSD of the peak area (n=3) was all < 1.5%, indicating that the method was repeatable. The purified VLP were diluted in serial and tested for linearity. The result suggested good linear relationship between the peak area of VLP or VLP aggregates and the protein concentration of the sample with R2 of 0.999 and 0.997, respectively. Thus, the method met the requirement for quantification and aggregate analysis. This method is accurate and efficient in in vitro quality evaluation and improvement of PCV2 vaccine.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Antibodies, Viral
		                        			;
		                        		
		                        			Capsid Proteins
		                        			;
		                        		
		                        			Chromatography, Gel
		                        			;
		                        		
		                        			Circoviridae Infections/prevention & control*
		                        			;
		                        		
		                        			Circovirus
		                        			;
		                        		
		                        			Lasers
		                        			;
		                        		
		                        			Swine
		                        			;
		                        		
		                        			Vaccines, Inactivated
		                        			;
		                        		
		                        			Vaccines, Virus-Like Particle
		                        			;
		                        		
		                        			Viral Vaccines
		                        			
		                        		
		                        	
6.NLRP3 inflammasome: an important therapeutic target for SARS-CoV-2 infection
Guorui LI ; Ning LI ; Qing GENG
Chinese Journal of Microbiology and Immunology 2022;42(3):171-177
		                        		
		                        			
		                        			SARS-CoV-2, the pathogen of the COVID-19 pandemic, causes serious damage to human health and social stability. In severe COVID-19 cases, the infection triggers cytokine storm, resulting in multi-organ excessive inflammatory responses and even failure, which eventually leads to death. Recent studies have shown the activation of nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3) inflammasome plays an essential role in the pathogenesis of COVID-19. SARS-CoV-2 can activate NLRP3 inflammasome through several pathways, thereby inducing the release of a large number of pro-inflammatory cytokines. This article reviews the activation of NLRP3 inflammasome caused by SARS-CoV-2 infection and the possible molecular mechanisms, and summarizes the progress in targeted inhibition of NLRP3 inflammation, aiming to provide a new strategy for the treatment of SARS-CoV-2 infection.
		                        		
		                        		
		                        		
		                        	
7.Genetic analysis of PYGL gene variants for a child with Glycogen storage disease VI.
Yucan ZHENG ; Guiping KONG ; Guorui HU ; Bixia ZHENG ; Mei LI
Chinese Journal of Medical Genetics 2022;39(2):209-212
		                        		
		                        			OBJECTIVE:
		                        			To explore the clinical features and genetic basis of a patient with glycogen storage disease type VI (GSD-VI).
		                        		
		                        			METHODS:
		                        			Clinical data of the patient was collected. Genomic DNA was extracted from peripheral blood samples of the proband and his parents. Genetic variants were detected by using whole exome sequencing. Candidate variants were verified by Sanger sequencing followed by bioinformatics analysis.
		                        		
		                        			RESULTS:
		                        			The proband presented fasting hypoglycemia, hepatomegaly, growth retardation, transaminitis, metabolic acidosis and hyperlactatemia. Liver biopsy indicated GSD. Novel compound heterozygous PYGL gene variants (c.2089A>G/c.158_160delACT) were detected in the proband. Compound heterozygosity was confirmed by Sanger sequencing of the patient's genomic DNA. Provean and MutationTaster predicted the two variants as deleterious and the variant sites are highly conserved.
		                        		
		                        			CONCLUSION
		                        			The compound heterozygous variants (c.2089A>G/c.158_160delACT) of PYGL gene probably underlay the GSD in the patient. The two novel variants have expanded the spectrum of PYGL gene variants and provided the basis for genetic counseling of the family.
		                        		
		                        		
		                        		
		                        			Child
		                        			;
		                        		
		                        			Family
		                        			;
		                        		
		                        			Genetic Testing
		                        			;
		                        		
		                        			Glycogen Storage Disease Type VI/genetics*
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Mutation
		                        			;
		                        		
		                        			Whole Exome Sequencing
		                        			
		                        		
		                        	
8.Electroacupuncture in the treatment of neurogenic urine retention through autophagy mediated by AMPK/mTOR pathway.
Zhengfei LI ; Ren ZHANG ; Guorui ZHAO ; Yao KUANG
Journal of Central South University(Medical Sciences) 2022;47(4):488-496
		                        		
		                        			OBJECTIVES:
		                        			Electroacupuncture can enhance autophagic flow, promote neuronal regeneration, axonal and myelin remodeling to achieve the protection of spinal cord injury, but its role in neurogenic urine retention is not completely clear. This study aims to investigate whether the mechanism of electroacupuncture in the treatment of neurogenic urine retention is through autophagy mediated by adenosine monophosphate activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway.
		                        		
		                        			METHODS:
		                        			A rat model of neurogenic urine retention after sacral spinal cord injury was established. The rats with successful model were randomly divided into a model group, an electroacupuncture group (electro-acupuncture for Ciliao, Zhongji, and Sanyinjiao by electronic stimulation, once a day, 20 min each time for 7 days), and an electroacupuncture+AMP-activated protein kinase (AMPK) inhibitor group (on the basis of the treatment of electroacupuncture group, 100 μg of AMPK inhibitor compound C was injected intramuscularly around the L2-3 intervertebral space on the 1st and 4th day). The normal group did not receive any treatment. The maximum bladder volume, bladder basal pressure, leak point pressure, and bladder compliance were recorded by multi-channel physiological recorder; the morphology of bladder tissue was observed by HE staining; autophagy was observed under transmission electron microscope; the expressions of LC3II and Beclin1 protein were observed by immunofluorescence staining; the protein levels of AMPK, phosphorylated-AMPK (p-AMPK), mTOR, phosphorylated-mTOR (p-mTOR), microtubule associated protein 1 light chain 3 (LC3) II and Beclin1 in bladder tissue were detected by Western blotting.
		                        		
		                        			RESULTS:
		                        			Compared with the normal group, the maximum bladder capacity, leak point pressure, bladder compliance, p-AMPK, LC3II, Beclin1 protein expressions in the bladder tissue of the model group increased, and the p-mTOR protein expressions were decreased (all P<0.05); compared with the model group, the maximum bladder capacity, bladder compliance, p-mTOR protein expression in the bladder tissue of the electroacupuncture group were decreased, and the p-AMPK, LC3II, and Beclin1 protein expressions were increased (all P<0.05); compared with the electroacupuncture group, the maximum bladder capacity, bladder compliance, p-mTOR protein expression in the bladder tissue of the electroacupuncture+AMPK inhibitor group were increased, the p-AMPK, LC3II, and Beclin1 protein expressions were decreased (all P<0.05). In the model group, the bladder became larger, with unclear and varying degrees of degeneration, severe tissue damage and autophagosome appeared; the bladder of the electroacupuncture group was smaller than that of the model group, and all levels were clearly visible with autophagy bodies; the layers were slightly disordered and damaged in the electroacupuncture + AMPK inhibitor group.
		                        		
		                        			CONCLUSIONS
		                        			Electroacupuncture can activate autophagy through AMPK/mTOR pathway, thereby reducing neurogenic urine retention caused by spinal cord injury.
		                        		
		                        		
		                        		
		                        			AMP-Activated Protein Kinases
		                        			;
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Autophagy
		                        			;
		                        		
		                        			Beclin-1
		                        			;
		                        		
		                        			Electroacupuncture
		                        			;
		                        		
		                        			Mammals
		                        			;
		                        		
		                        			Rats
		                        			;
		                        		
		                        			Rats, Sprague-Dawley
		                        			;
		                        		
		                        			Spinal Cord Injuries
		                        			;
		                        		
		                        			TOR Serine-Threonine Kinases
		                        			
		                        		
		                        	
9.Characteristics of polysomnography in depression patients complicating moderate-to-severe obstructive sleep apnea hypopnea syndrome
Gang YE ; Zhe LI ; Wen PAN ; Shigeng GAO ; Yingying SHEN ; Ying LIU ; Ming YIN ; Guorui LIU ; Yu ZHAO ; Jianhong SHEN ; Xiangdong DU
Sichuan Mental Health 2021;34(5):424-428
		                        		
		                        			
		                        			ObjectiveTo investigate the characteristics of polysomnography (PSG) in depression patients complicating moderate-to-severe obstructive sleep apnea hypopnea syndrome (OSAHS). MethodsA retrospective analysis was conducted on the outpatients, inpatients and physical examination population who completed overnight PSG monitoring in the sleep medicine center of Suzhou Guangji Hospital from December 2017 to October 2019. Four groups of subjects were finally enrolled, including depression patients with moderate-to-severe OSAHS (n=31), depression patients without OSAHS (n=79), moderate-to-severe OSAHS patients (n=96) and normal control group (n=32). The sleep process related indicators (total sleep time, sleep latency, number of awakenings), sleep structure related indicators (N1, N2, N3, percentage of REM sleep, REM latency, REM sleep duration), sleep-related respiratory variables (oxygen reduction index) and other polysomnographic parameters of the four groups were compared. ResultsIn terms of sleep process, the total sleep time, sleep latency and number of awakenings yielded significant differences among the four groups (F=2.874, 3.959, 12.291, P<0.05 or 0.01). In terms of sleep structure, the percentage of total sleep time in N2 and N3 stages demonstrated significant differences among the four groups (F=13.885, 48.013, P<0.01). The REM latency, REM sleep duration and percentage of REM sleep manifested significant differences among the four groups (F=41.492, 11.827, 10.552, P<0.01). In terms of sleep-related respiratory variables, the oxygen reduction index exhibited significant differences among the four groups (F=170.585, P<0.05). ConclusionDepression patients complicating moderate-to-severe OSAHS suffer from severe sleep process and structural disturbances, accompanied by quite frequent and severe sleep-related respiratory events. 
		                        		
		                        		
		                        		
		                        	
10.Clinicalefficiencyofendovascularinterventionaltherapyforthesolitarykidneywithrenalaneurysm
Fangzheng LI ; Jianzhuang REN ; Wenguang ZHANG ; Xuhua DUAN ; Jiaxing WANG ; Li WAN ; Linhui XU ; Guorui ZHAO ; Pengfei CHEN
Journal of Practical Radiology 2019;35(7):1128-1131
		                        		
		                        			
		                        			Objective Toanalyzetheclinicalefficiencyofendovascularinterventionaltherapyinthetreatmentofsolitarykidney withrenalaneurysm.Methods FromJune2014toDecember2017,7casesofsolitarykidneypatientscombinedwithrenalaneurysm werecollected,including4casesofrenalaneurysmlocatedinthemainrenalartery,1caseinrenalsegmentalarteryand2casesinthe renalparenchyma.4casesofmainrenalarteryaneurysmsweretreatedwithstentimplantation;1caseofrenalsegmentalarteryaneurysm was treatedwithstent-assistedcoilembolization;2casesofrenalparenchymaaneurysm wererespectivelytreatedwithpolyvinylalcohol (PVA)embolizationandPVA withcoilembolization.After1month,3months,6monthsandevery6months,therenaldopplerexamination,blood routine,urineroutine,renalfunctionandotherbiochemicalexaminationswerereviewed,andrenalarterialCTAorrenalangiographywereperformed whennecessary.Results Thesuccessrateofoperationwas100%in7solitarykidneypatientswithrenalaneurysm,andtheaneurysmswere graduallyreducedafteroperationwithoutseriousadversereactionsandcomplications.Norecanalizationorrecurrenceofrenalaneurysmwasfound duringthefollow-up,andtherenalfunctionwasbetterthanbefore.Conclusion Endovascularinterventionaltherapyinthetreatment ofsolitarykidneywithrenalaneurysmisminimallyinvasiveandlessrisky.Anditismoretargetedandflexibleforthetreatmentof solitarykidneywithdifferenttypesofrenalaneurysm.
		                        		
		                        		
		                        		
		                        	
            
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