Ophiopogon japonicus, a precious medicinal plant endemic to Zhejiang Province. Its tuberous roots are rich in bioactive components such as flavonoids, possessing anti-inflammatory, antioxidant, and immunomodulatory properties. To elucidate the impact of cadmium (Cd) stress on the accumulation and biosynthetic pathway of flavonoids in O. japonicus, this study exposed O. japonicus to different concentrations of Cd stress and explored the changes through integrated transcriptomics and metabolomics analysis. The results demonstrated that Cd stress (1 mg/L and 10 mg/L) significantly increased the content of flavonoids in O. japonicus in a concentration-dependent manner. The metabolomics analysis revealed a total of 110 flavonoids including flavones, flavanols, flavonols, flavone and flavonol derivatives, flavanones, isoflavonoids, chalcones and dihydrochalcones, and anthocyanins in O. japonicus, among which flavones, flavonols, flavone and flavonol derivatives, and anthocyanins increased under Cd stress. The transcriptomics analysis identified several key flavonoid biosynthesis-associated genes with up-regulated expression under Cd stress, including 14 genes encoding 4-coumarate CoA ligase (4CL), 2 genes encoding chalcone isomerase (CHI), and 14 genes encoding phenylalanine ammonia lyase (PAL). The gene-metabolite regulatory network indicated significant positive correlations of 4CL (Cluster-21637.5012, Cluster-21637.90648, and Cluster-21637.62637) and CHI (Cluster-21637.111909 and Cluster-21637.123300) with flavonoid metabolites, suggesting that these genes promoted the synthesis of specific flavonoid metabolites, which led to the accumulation of total flavonoids under Cd stress. These findings provide theoretical support for the cultivation and utilization of medicinal plants in Cd-contaminated environments and offered new perspectives for studying plant responses to heavy metal stress.
Cadmium/toxicity* ; Flavonoids/biosynthesis* ; Metabolomics ; Ophiopogon/drug effects* ; Stress, Physiological ; Transcriptome ; Gene Expression Profiling ; Gene Expression Regulation, Plant

Soil cadmium (Cd) pollution is one of the major environmental problems globally. Ophiopogon japonicus, a multifunctional plant extensively used in traditional Chinese medicine, has demonstrated potential in environmental remediation. This study investigated the Cd accumulation pattern of O. japonicus under cadmium stress and identified the heavy metal ATPase (HMA) family members in this plant. Our results demonstrated that O. japonicus exhibited a Cd enrichment factor (EF) of 2.75, demonstrating strong potential for soil Cd pollution remediation. Nine heavy metal ATPase (HMA) members of P1B-ATPases were successfully identified from the transcriptome data of O. japonicus, with OjHMA1-OjHMA6 classified as the Zn/Co/Cd/Pb-ATPases and OjHMA7-OjHMA9 as the Cu/Ag-ATPases. The expression levels of OjHMA1, OjHMA2, OjHMA3, and OjHMA7 were significantly up-regulated under Cd stress, highlighting their crucial roles in cadmium ion absorption and transport. The topological analysis revealed that these proteins possessed characteristic transmembrane (TM) segments of the family, along with functional A, P, and N domains involved in regulating ion absorption and release. Metal ion-binding sites (M4, M5, and M6) existed on the TM segments. Based on the number of transmembrane domains and the residues at metal ion-binding sites, the plant HMA family members were categorized into three subgroups: P1B-1 ATPases, P1B-2 ATPases, and P1B-4 ATPases. Specifically, the P1B-1 ATPase subgroup included the motifs TM4(CPC), TM5(YN[X]₄P), and TM6(M[XX]SS); the P1B-2 ATPase subgroup featured the motifs TM4(CPC), TM5(K), and TM6(DKTGT); the P1B-4 ATPase subgroup contained the motifs TM4(SPC) and TM6(HE[X]GT), all of which were critical for protein functions. Molecular docking results revealed the importance of conserved sequences such as CPC/SPC, DKTGT, and HE[X]GT in metal ion coordination and stabilization. These findings provide potential molecular targets for enhancing Cd uptake and tolerance of O. japonicus by genetic engineering and lay a theoretical foundation for developing new cultivars with high Cd accumulation capacity.
Cadmium/metabolism* ; Adenosine Triphosphatases/metabolism* ; Ophiopogon/drug effects* ; Soil Pollutants/toxicity* ; Plant Proteins/metabolism* ; Stress, Physiological ; Multigene Family ; Gene Expression Regulation, Plant

Objective To investigate the regulations of lymphatic metastasis of rectal carcinoma which can provide help of clinical diagnosis and treatments.Methods A retrospective analysis was performed of 222 patients with rectal carcinoma who underwent surgical resection from November 2006 to January 2012.Summarize the relevance of each factor and lymph node metastasis by logistic regression analysis.Results (1) Gender and lymph node metastasis has no significant correlation (x2 =0.000,P ＞ 0.05).(2) Lymph node metastasis rate reduced with increasing age and the difference was statistically significant (x2 =5.001,P ＜ 0.05).(3) There is no significant difference between tumor location and lymph node metastasis rate (x2 =0.052,P ＞ 0.05),when the distance between anal rectal cancer and the anus was closer,the tendency of lymph node metastasis was higher,and there has the obvious correlation with lymph node metastasis rate of its size,2.5 to 5.5 cm group and more than 5.5 cm lymph node metastasis rate was significantly higher than less than 2.5 cm group with a significant difference (x2 =2.267,P ＜ 0.05).(4) Tumor invasion of the intestinal circumference was closely related with lymph node metastasis rate (x2 =2.323,P ＜0.01).(5) Lymph node metastasis rate was closely related to the depth of invasion,metastasis rate increased with the depth of invasion increasing(x2 =2.277,P ＜0.05).(6) Lymph node metastasis was significantly higher with the decrease of the degree of tumor differentiation (x2 =4.322,P ＜ 0.01).Logistic regression analysis showed that the degree of tumor differentiation,infiltration and invasion of intestinal circumference diameter lymph node metastasis influential,OR (95 ％ CI) was 5.562 (0.105-0.444),2.178 (0.144-0.971),1.563 (0.038-8.613) respectively (P ＜0.01).Conclusion The lymphatic metastasis of rectal carcinoma is related to age,tumor size,intestinal circumference invasion,depth of tumor invasion and the differentiating degree of the tumor,and the differentiating degree is the major factor.

Objective To evaluate whether the gastroesophageal reflux disease (GERD) questionnaire (GerdQ) symptom score could reflect the acid exposure indicated by 24-hour esophageal pH monitoring in patients with GERD, and to further validate the clinical application value of GerdQ.Methods From November 2008 to March 2010, 134 GERD patients from outpatient department with heartburn and acid reflux symptoms were enrolled.All the patients had undergone endoscopy examination, 24-hour esophageal pH monitoring and GerdQ scoring.According to 24-hour esophageal pH monitoring result, acid exposure group and none acid exposure group was divided by the percentage of time that pH was lower than 4 over 4.2% or DeMeester score over 14.72, the difference of GerdQ score was assessed.The receiver operating characteristic (ROC) curve of GerdQ score was drawn according to DeMeester score and the percentage of time that pH was lower than 4 as the gold standard.With SPSS 17.0 statistical software, the ratio was compared with the u test and Chi-square test, comparison between groups with t test.Results The BMI of acid exposure group was higher than that of none acid exposure group(t= 2.365, P= 0.019).The GerdQ score of acid exposure group (8.51± 2.34) was significantly higher than that of none acid exposure group (6.67 ± 2.79), the difference was statistically significant (t = 4.002, P = 0.000) .ROC curve was drawn according to DeMeester score over 14.72 as gold standard; the area under curve (AUC) was 0.70.When the critical value of GerdQ score was 7.5, Youden index was up to 0.3365, the sensitivity of diagnosis was 0.6909 and specificity was 0.6454.The percentage of time that PH was lower than 4 over 4.2% as the gold standard, the AUC of GerdQ score was 0.663.When the critical value of GerdQ score was 7.5, Youden index was up to 0.2790, the sensitivity of diagnosis was 0.6600 and specificity was 0.6190.Conclusions GerdQ score greater than 7.5 can better reflect acid exposure at the lower esophageal.There is stronger association between GerdQ score with the DeMeester acid exposure score over 14.72 than with the percentage of time that pH lower than 4 over 4.2% as the critical value for acid exposure.Obesity may aggravate the transient lower esophageal sphincter relaxation leading to occurrence of GERD symptoms.