ObjectiveTo explore the effects of genetic variation of obesity-related biological pathways and gene-obesity interactions on the incidence of gastric cancer, so as to better understand the pathogenesis of gastric cancer and help identify high-risk populations for individualized prevention of gastric cancer. MethodsA case-control study based on the Shanghai Suburban Adult Cohort and Biobank study (SSACB) was conducted on the cases with gastric cancer. A total of 267 cases with gastric cancer and 267 healthy controls matched 1∶1 by age and gender using propensity score were included in the study. After genome-wide genotyping, quality control and imputation, 19 250 single nucleotide polymorphism (SNP) sites from 115 genes in 4 obesity-related biological pathways were extracted. Univariate and multivariate logistic regression analyses were used to evaluate the association between these SNP sites and the risk of gastric cancer, and false positive report probability (FPRP) was used for multiple test correction.Data from Biobank Japan (BBJ) and FinnGen public accessible databases were used to validate significant SNP sites. For validated sites, expression quantitative trait loci (eQTL) analysis and differentially expressed genes analysis were further performed. Additive and multiplicative interactions were used to evaluate the gene-obesity interactions on the incidence of gastric cancer. Additive interaction evaluation indicators included relative excess risk due to interaction (RERI), attributable proportion due to interaction (AP) and synergy index (SI), while multiplicative interaction evaluation indicators include OR_GxE and P_inter. ResultsA total of 41 SNP sites were significantly associated with the onset of gastric cancer (P_adj<0.05, FPRP_0.1<0.1), among which 7 groups of haplotype blocks were formed. ACACB/ rs2268401 [SSACB: P=0.005, BBJ: P=0.049], HRAS/ rs12785860 (SSACB: P<0.001, FinnGen: P=0.045), and PTPN1/ rs6095985 (SSACB: P<0.001, FinnGen: P=0.023) were significantly associated with the risk of gastric cancer after validation in different populations. Among which, the G allele of HRAS/ rs12785860 was correlated with the downregulation of HRAS mRNA expression (P<0.001), and the expression level of HRAS in gastric cancer tissues was higher than that in adjacent normal tissues (P<0.001). Additionaly, JAK1/rs11208559 showed a positive additive interaction with waist circumstance (WC) on the risk of gastric cancer [RERI=2.29(0.06~4.53), AP=0.57(0.23~0.90), SI=4.03(2.20~5.87)]. ConclusionObesity-related biological pathway SNP sites and their haplotypes are associated with the risk of gastric cancer, suggesting that genetic variations in obesity pathways may affect gastric cancer. The HRAS/ rs12785860 is significantly associated with downregulation of HRAS gene expression, which may serve as a potential genetic marker for gastric cancer. JAK1/rs11208559 interacts with obesity additively on the risk of gastric cancer. Individuals with GC+CC genotypes and pre-central or central obesity have an increased risk of gastric cancer, providing clues and evidences for individualized prevention of gastric cancer.

Gastric cancer (GC) is characterized by high morbidity and mortality rates. Chinese agarwood comprises the resin-containing wood of Aquilaria sinensis (Lour.) Gilg., traditionally utilized for treating asthma, cardiac ischemia, and tumors. However, comprehensive research regarding its anti-GC effects and underlying mechanisms remains limited. In this study, Chinese agarwood petroleum ether extract (CAPEE) demonstrated potent cytotoxicity against human GC cells, with half maximal inhibitory concentration (IC₅₀) values for AGS, HGC27, and MGC803 cells of 2.89, 2.46, and 2.37 μg·mL^-1, respectively, at 48 h. CAPEE significantly induced apoptosis in these GC cells, with B-cell lymphoma-2 (BCL-2) associated X protein (BAX)/BCL-2 antagonist killer 1 (BAK) likely mediating CAPEE-induced apoptosis. Furthermore, CAPEE induced G₀/G₁ phase cell cycle arrest in human GC cells via activation of the deoxyribonucleic acid (DNA) damage-p21-cyclin D1/cyclin-dependent kinase 4 (CDK4) signaling axis, and increased Fe²⁺, lipid peroxides and reactive oxygen species (ROS) levels, thereby inducing ferroptosis. Ribonucleic acid (RNA) sequencing, real-time quantitative polymerase chain reaction (RT-qPCR), and Western blotting analyses revealed CAPEE-mediated upregulation of heme oxygenase-1 (HO-1) in human GC cells. RNA interference studies demonstrated that HO-1 knockdown reduced CAPEE sensitivity and inhibited CAPEE-induced ferroptosis in human GC cells. Additionally, CAPEE administration exhibited robust in vivo anti-GC activity without significant toxicity in nude mice while inhibiting tumor cell growth and promoting apoptosis in tumor tissues. These findings indicate that CAPEE suppresses human GC cell growth through upregulation of the DNA damage-p21-cyclin D1/CDK4 signaling axis and HO-1-mediated ferroptosis, suggesting its potential as a candidate drug for GC treatment.
Animals ; Humans ; Mice ; Antineoplastic Agents, Phytogenic ; Apoptosis/drug effects* ; Cell Line, Tumor ; Cyclin D1/genetics* ; Cyclin-Dependent Kinase 4/genetics* ; DNA Damage/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Ferroptosis/drug effects* ; G1 Phase Cell Cycle Checkpoints/drug effects* ; Heme Oxygenase-1/genetics* ; Mice, Inbred BALB C ; Mice, Nude ; Plant Extracts/pharmacology* ; Stomach Neoplasms/physiopathology* ; Thymelaeaceae/chemistry* ; Up-Regulation/drug effects*

BACKGROUND: Lung cancer is one of the malignant cancers with the highest incidence rate, and it is important to identify the factors contributing to lung cancer carcinogenesis for prevention. Lifestyle and genetic factors play important roles in cancer development, however the impact of dietary factors, such as soy product intake, on lung cancer risk remains inadequately understood. This study aims to explore the associations between soy product intake, genetic risk, and lung cancer incidence, and validate the consistent effects of soy product intake in European populations, thereby providing new insights for lung cancer prevention. METHODS: Utilizing the Shanghai Suburban Adult Cohort and Biobank (SSACB) (n=66,311), Cox proportional hazards model was adopted to assess the association between soy product intake and lung cancer incidents, followed by subgroup analyses stratified by gender, smoking status, and pathological types of lung cancer. The UK Biobank (UKB) was used for validation of the effect of soy product intake on lung cancer. To investigate the association between genetic factors and lung cancer, in addition to previously reported loci, we incorporated newly identified loci from two independent studies in Southeast China: a nested case-control population from the SSACB cohort (433 cases/650 controls) and a case-control study from the Shanghai Cancer Center-Taizhou cohort (1359 cases/1359 controls). Meta-analysis and Linkage disequilibrium clumping (LD clumping) of the association results identified 23 loci for polygenic risk score (PRS) construction. Subsequently, conditional Logistic regression model was used to assess the association between genetic risk and lung cancer. RESULTS: In SSACB cohort, after adjusting for age, gender, smoking, chronic bronchitis, body mass index (BMI), vegetable intake and red meat intake, sufficient soy product intake was significantly associated with a reduced risk of lung cancer [hazard ratio (HR)=0.60, 95%CI: 0.47-0.77, Padj=6.69E-05], an effect that was consistent in males and females, smokers and non-smokers. In UKB, although the association did not reach statistical significance, a protective trend against lung cancer was also observed (HR=0.76, 95%CI: 0.55-1.06, Padj=0.10). In the nested case-control population within SSACB, a PRS score generated in the Chinese population was significantly correlated with lung cancer risk. After adjustment of age, gender, smoking, chronic bronchitis, and soy product intake, the high-PRS group had a 1.88 times higher risk of lung cancer compared to the low-PRS group (Padj=1.84E-03). CONCLUSIONS The prospective cohort study found that adequate intake of soy products was significantly associated with a reduced risk of lung cancer, while a high PRS is a risk factor for lung cancer development. Integrating soy product intake and PRS into traditional epidemiological risk factor prediction will guide personalized lung cancer prevention and high-risk population stratification.
Humans ; Lung Neoplasms/etiology* ; Male ; Female ; China/epidemiology* ; Middle Aged ; Adult ; Aged ; Prospective Studies ; Biological Specimen Banks ; Risk Factors ; Case-Control Studies ; Cohort Studies

Objective:To explore the effects of carcinoma-associated fibroblasts (CAF) in gastric cancer microenvironment on the proliferation of gastric cancer organoids and the sensitivity to chemotherapeutic drugs, as well as the underlying mechanisms.Methods:Tissue specimens of 6 patients with gastric cancer who underwent surgical resection in Fujian Cancer Hospital from June 2022 to January 2024 were collected from the biobank. Organoids derived from gastric cancer and isolated tumor tissue CAF and paracancerous tissue normal fibroblasts (NF) were constructed by the primary tissue culture method. The morphology of organoids and fibroblasts was observed under microscope. Immunohistochemistry (IHC) method was used to detect the expressions of gastric cancer markers in organoids. The expression levels of fibroblast related markers were detected by using immunofluorescence, reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blotting methods. A co-culture model of gastric cancer organoids and fibroblasts was constructed by using the Transwell chamber and then was divided into 3 groups: NF and gastric cancer organoids co-culture group (NF co-culture group), CAF and gastric cancer organoids co-culture group (CAF co-culture group), and gastric cancer organoid separate culture group (separate culture group). Different concentrations of oxaliplatin, paclitaxel, and 5-fluorouracil (5-FU) were applied to the co-culture system of fibroblasts and gastric cancer organoids. The morphology of gastric cancer organoids in each group was observed under microscope, and the relative viability of the organoids was detected by using the CellTiter-Glo 3D luminescence method. CAF and NF of 3 patients who successfully constructed organoids were collected for transcriptome sequencing, and the differentially expressed genes (with |log 2 fold change| > 0 and P < 0.05) between CAF and NF were analyzed. Pathway enrichment analysis of the upregulated differentially expressed genes in CAF was performed by using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Results:The organoids of 4 patients were successfully constructed, among which 3 organoids with relatively consistent results and covering the heterogeneity of the disease were performed for further analysis. After HE staining, microscopic observation showed that the morphological characteristics of the organoids were homogeneous with those of gastric cancer tissues. IHC method detection showed that both organoids and primary tumor tissues both expressed gastric cancer markers CDX2, Villin, and CK20. Inverted microscopic observation revealed that NF was morphologically similar to CAF, but NF had fewer cytoplasmic protrusions. RT-qPCR method, immunofluorescence and Western blotting detections showed that the mRNA and protein expression levels of α-smooth muscle actin (α-SMA), fibroblast activation protein (FAP) and Vimentin in CAF were all higher than those in NF, and the differences were statistically significant (all P < 0.05). Inverted microscope observation showed that after co-cultured for 96 h, the size of organoids in CAF co-culture group [(308±61) μm] was larger than that in NF co-culture group [(155±33) μm] and separate culture group [(91±17) μm], and the differences were statistically significant (all P < 0.05). CellTiter-Glo 3D luminescence assay showed that the relative viability of organoids in the CAF co-culture group was enhanced after 96 h of co-culture compared with NF co-culture group and separate culture group, and the differences were statistically significant (all P < 0.05); there was no statistically significant difference in the relative viability between NF co-culture group and separate culture group ( P > 0.05). With the increase of the concentrations of oxaliplatin, 5-FU and paclitaxel, the relative activity of organoids after drug effect for 96 h in the 3 groups gradually decreased. The relative viability of the organoids in CAF co-culture groups treated with 6.25, 12.5, 25, 50, and 100 μmol/L oxaliplatin, 12.5, 25, 50, and 100 μmol/L 5-FU, and 0.625, 1.25, 2.5, and 5 μmol/L paclitaxel was higher than that of the other 2 groups, and the differences were statistically significant (all P < 0.05). Transcriptome sequencing analysis revealed that compared with NF, there were 893 differentially expressed genes with upregulated mRNA expression and 424 differentially expressed genes with downregulated mRNA expression in CAF. KEGG pathway enrichment analysis showed that the upregulated differentially expressed genes were mainly involved in the calcium signaling pathway. Conclusions:CAF can promote the proliferation ability of gastric cancer organoids and decrease the sensitivity to chemotherapy drugs, which may be related to the calcium signaling pathway.

Objective:To adapt the Meaning in Life in Children Questionnaire(MIL-CQ)into a Chinese ver-sion and examine its validity and reliability.Methods:A total of 1 649 primary school students were recruited and divided into 4 samples.Sample 1(n=527)was used for item analysis and exploratory factor analysis.Sample 2(n=503)was used for confirmatory factor analysis(CFA).In sample 3(n=512),criterion validity was assessed u-sing the Chinese version of Satisfaction with Life Scale(C-SWLS),Williams Creativity Assessment Packet(CAP),Quality of Life Scale for Children and Adolescents(QLSCA),and Strength and Difficulty Questionnaire(SDQ),followed by a second confirmatory factor analysis.Sample 1 and Sample 2 were combined for internal con-sistency reliability analysis;Sample 4(n=107)was used for test-retest reliability analysis.Results:The exploratory factor analysis showed that the Chinese version of the MIL-CQ contained 18 items,including three factors(attitude,creativity,and experience).Both CFAs showed excellent fit(CFI=0.95/0.96,TLI=0.94/0.96,SRMR=0.04,RMSEA=0.04,x2/df=1.60/1.71,Ps＜0.001).The total scores and subscale scores of the Chinese version of MIL-CQ were positively correlated with the scores of C-SWLS,CAP,QLSCA,and the prosocial behavior section of the SDQ(r=0.17-0.58,Ps＜0.01),and negatively correlated with the difficulty section of the SDQ(r=-0.06--0.39,Ps＜0.01).The Cronbach's α coefficients for the total scale and the three subscales ranged from 0.64 to 0.83,and the test-retest reliability(ICC)over a 1-month interval ranged from 0.69 to 0.77.Conclusion:The Chi-nese version of the Meaning in Life in Children Questionnaire has good validity and reliability for assessing mean-ing in life among children.

Objective:To observe the therapeutic effects of electroacupuncture(EA)stimulation on chronic pain and associated negative emotions,and to investigate the activity changes of glutamatergic neurons in the primary motor cortex(M1)following EA stimulation.Methods:Male C57BL/6J mice were randomly divided into four groups:sham surgery(Sham),pure electroacupuncture stimulation(EA),nerve injury(SNI)and electroacupuncture treatment of nerve injury(SNI+EA).Thirty days after the SNI model establishment,EA stimulation was administered bilaterally to the Zusanli(ST36)acupoints in mice.von Frey filaments and Hargreaves heat sensitivity testing were used to detect mechanical and thermal hyperalgesia.The elevated plus maze test was conducted to assess the impact on anxiety-like behaviors in mice.Dual immunofluorescence staining was employed to observe changes in c-Fos expression in M1 gluta-matergic neurons.Results:As compared to Sham group,SNI-treated mice exhibited significant mechanical and thermal hyperalgesia in bilateral hindpaws at 30 days post-modeling(P＜0.01)and displayed obvious anxiety-like behaviors(P＜0.01).The SNI+EA group showed significant relief in pain and anxiety-like behaviors(P＜0.01).c-Fos expression in M1 glutamatergic neurons was significantly decreased in SNI mice.Conversely,after electroacupuncture(EA)treatment,compared to the SNI group,M1 glutamatergic neurons in the SNI+EA group showed significantly in-creased c-Fos expression(P＜0.01).Conclusion:Electroacupuncture significantly alleviated chronic pain and associ-ated anxiety-like behaviors induced by SNI,which might involve the activation of glutamatergic neurons in the M1 in this process.

Objective:To adapt the Meaning in Life in Children Questionnaire(MIL-CQ)into a Chinese ver-sion and examine its validity and reliability.Methods:A total of 1 649 primary school students were recruited and divided into 4 samples.Sample 1(n=527)was used for item analysis and exploratory factor analysis.Sample 2(n=503)was used for confirmatory factor analysis(CFA).In sample 3(n=512),criterion validity was assessed u-sing the Chinese version of Satisfaction with Life Scale(C-SWLS),Williams Creativity Assessment Packet(CAP),Quality of Life Scale for Children and Adolescents(QLSCA),and Strength and Difficulty Questionnaire(SDQ),followed by a second confirmatory factor analysis.Sample 1 and Sample 2 were combined for internal con-sistency reliability analysis;Sample 4(n=107)was used for test-retest reliability analysis.Results:The exploratory factor analysis showed that the Chinese version of the MIL-CQ contained 18 items,including three factors(attitude,creativity,and experience).Both CFAs showed excellent fit(CFI=0.95/0.96,TLI=0.94/0.96,SRMR=0.04,RMSEA=0.04,x2/df=1.60/1.71,Ps＜0.001).The total scores and subscale scores of the Chinese version of MIL-CQ were positively correlated with the scores of C-SWLS,CAP,QLSCA,and the prosocial behavior section of the SDQ(r=0.17-0.58,Ps＜0.01),and negatively correlated with the difficulty section of the SDQ(r=-0.06--0.39,Ps＜0.01).The Cronbach's α coefficients for the total scale and the three subscales ranged from 0.64 to 0.83,and the test-retest reliability(ICC)over a 1-month interval ranged from 0.69 to 0.77.Conclusion:The Chi-nese version of the Meaning in Life in Children Questionnaire has good validity and reliability for assessing mean-ing in life among children.

Objective:To observe the therapeutic effects of electroacupuncture(EA)stimulation on chronic pain and associated negative emotions,and to investigate the activity changes of glutamatergic neurons in the primary motor cortex(M1)following EA stimulation.Methods:Male C57BL/6J mice were randomly divided into four groups:sham surgery(Sham),pure electroacupuncture stimulation(EA),nerve injury(SNI)and electroacupuncture treatment of nerve injury(SNI+EA).Thirty days after the SNI model establishment,EA stimulation was administered bilaterally to the Zusanli(ST36)acupoints in mice.von Frey filaments and Hargreaves heat sensitivity testing were used to detect mechanical and thermal hyperalgesia.The elevated plus maze test was conducted to assess the impact on anxiety-like behaviors in mice.Dual immunofluorescence staining was employed to observe changes in c-Fos expression in M1 gluta-matergic neurons.Results:As compared to Sham group,SNI-treated mice exhibited significant mechanical and thermal hyperalgesia in bilateral hindpaws at 30 days post-modeling(P＜0.01)and displayed obvious anxiety-like behaviors(P＜0.01).The SNI+EA group showed significant relief in pain and anxiety-like behaviors(P＜0.01).c-Fos expression in M1 glutamatergic neurons was significantly decreased in SNI mice.Conversely,after electroacupuncture(EA)treatment,compared to the SNI group,M1 glutamatergic neurons in the SNI+EA group showed significantly in-creased c-Fos expression(P＜0.01).Conclusion:Electroacupuncture significantly alleviated chronic pain and associ-ated anxiety-like behaviors induced by SNI,which might involve the activation of glutamatergic neurons in the M1 in this process.

Objective:To explore the effects of carcinoma-associated fibroblasts (CAF) in gastric cancer microenvironment on the proliferation of gastric cancer organoids and the sensitivity to chemotherapeutic drugs, as well as the underlying mechanisms.Methods:Tissue specimens of 6 patients with gastric cancer who underwent surgical resection in Fujian Cancer Hospital from June 2022 to January 2024 were collected from the biobank. Organoids derived from gastric cancer and isolated tumor tissue CAF and paracancerous tissue normal fibroblasts (NF) were constructed by the primary tissue culture method. The morphology of organoids and fibroblasts was observed under microscope. Immunohistochemistry (IHC) method was used to detect the expressions of gastric cancer markers in organoids. The expression levels of fibroblast related markers were detected by using immunofluorescence, reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blotting methods. A co-culture model of gastric cancer organoids and fibroblasts was constructed by using the Transwell chamber and then was divided into 3 groups: NF and gastric cancer organoids co-culture group (NF co-culture group), CAF and gastric cancer organoids co-culture group (CAF co-culture group), and gastric cancer organoid separate culture group (separate culture group). Different concentrations of oxaliplatin, paclitaxel, and 5-fluorouracil (5-FU) were applied to the co-culture system of fibroblasts and gastric cancer organoids. The morphology of gastric cancer organoids in each group was observed under microscope, and the relative viability of the organoids was detected by using the CellTiter-Glo 3D luminescence method. CAF and NF of 3 patients who successfully constructed organoids were collected for transcriptome sequencing, and the differentially expressed genes (with |log 2 fold change| > 0 and P < 0.05) between CAF and NF were analyzed. Pathway enrichment analysis of the upregulated differentially expressed genes in CAF was performed by using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Results:The organoids of 4 patients were successfully constructed, among which 3 organoids with relatively consistent results and covering the heterogeneity of the disease were performed for further analysis. After HE staining, microscopic observation showed that the morphological characteristics of the organoids were homogeneous with those of gastric cancer tissues. IHC method detection showed that both organoids and primary tumor tissues both expressed gastric cancer markers CDX2, Villin, and CK20. Inverted microscopic observation revealed that NF was morphologically similar to CAF, but NF had fewer cytoplasmic protrusions. RT-qPCR method, immunofluorescence and Western blotting detections showed that the mRNA and protein expression levels of α-smooth muscle actin (α-SMA), fibroblast activation protein (FAP) and Vimentin in CAF were all higher than those in NF, and the differences were statistically significant (all P < 0.05). Inverted microscope observation showed that after co-cultured for 96 h, the size of organoids in CAF co-culture group [(308±61) μm] was larger than that in NF co-culture group [(155±33) μm] and separate culture group [(91±17) μm], and the differences were statistically significant (all P < 0.05). CellTiter-Glo 3D luminescence assay showed that the relative viability of organoids in the CAF co-culture group was enhanced after 96 h of co-culture compared with NF co-culture group and separate culture group, and the differences were statistically significant (all P < 0.05); there was no statistically significant difference in the relative viability between NF co-culture group and separate culture group ( P > 0.05). With the increase of the concentrations of oxaliplatin, 5-FU and paclitaxel, the relative activity of organoids after drug effect for 96 h in the 3 groups gradually decreased. The relative viability of the organoids in CAF co-culture groups treated with 6.25, 12.5, 25, 50, and 100 μmol/L oxaliplatin, 12.5, 25, 50, and 100 μmol/L 5-FU, and 0.625, 1.25, 2.5, and 5 μmol/L paclitaxel was higher than that of the other 2 groups, and the differences were statistically significant (all P < 0.05). Transcriptome sequencing analysis revealed that compared with NF, there were 893 differentially expressed genes with upregulated mRNA expression and 424 differentially expressed genes with downregulated mRNA expression in CAF. KEGG pathway enrichment analysis showed that the upregulated differentially expressed genes were mainly involved in the calcium signaling pathway. Conclusions:CAF can promote the proliferation ability of gastric cancer organoids and decrease the sensitivity to chemotherapy drugs, which may be related to the calcium signaling pathway.

Objective:To evaluate the endometrial implantation window in patients with recurrent implantation failure using endometrial receptive array(ERA)sequencing or endometrial histological detection methods,and to explore the effectiveness and cost-effectiveness analysis of two technologies for improving clinical outcomes in such patients.Methods:A retrospective cohort study was conducted on clinical data of 125 patients diagnosed with repeated implantation failure in Gansu Maternal and Child Health Hospital from January 2018 to December 2022.According to whether endometrial receptivity testing was accepted and different detection techniques were used,they were divided into a control group(n=36),a genomic group(n=35),and a histological group(n=54).The clinical data and pregnancy outcomes of the three groups were compared.Results:①The results of one-way ANOVA showed that the embryo implantation rate in the genomic group and histological group was significantly higher than that in the control group,and the difference was statistically significant(P＜0.05).There was no sta-tistically significant difference in embryo implantation rate between genomic and histological groups(P=0.48).②There was no statistically significant difference in clinical pregnancy rate and live birth rate among the three groups(P＞0.05).③Log rank test showed:The time for 50％of patients to reach live labor was significantly shorter than that of the control group,and the difference was statistically significant(P＜0.05);There was no sta-tistically significant difference in the time to live birth in 50％of patients between the genomic and histological groups of 50％of patients(P＞0.05).④The average number of embryos transferred in the control group was significantly higher than that in the genomic and histological groups,with statistical significance(P＜0.05).The cost of genomic patients was significantly higher than that of histology group,and the difference was statistically significant(P＜0.05).Conclusions:①Endometrial implantation window detection is feasible for patients with re-peated implantation failure,which can effectively shorten the time to live birth and reduce the number of transplan-ted embryos;②Both ERA sequencing and endometrial histology detection have limitations as methods to evaluate endometrial implantation window,and it is not clear which detection method has more advantages in accuracy and practicability.