A qualitative analysis by high-performance liquid chromatography coupled with time-of-flight mass spectrometry (HPLC-QTOF-MS/MS) was performed for the identification of main constituents in Gentiana veitchiorum. High-performance liquid chromatography (HPLC) was developed for the quantification of seven major components, including loganic acid (1), swertiamarin (2), gentiopicroside (3), sweroside (4), isoorientin (5), isoscoparin (6), and gentiournoside A (7). A total of 42 compounds, including 31 flavonoids, and 11 Iridoids, were identified based on their retention behaviors, and MS fragment information. Furthermore, regression equations for these seven chemical components were established, with good linear relationships (r2 > 0.9999), and the sample recovery rate was 97.02%-103.08%. This method was successfully applied for simultaneous determination of seven components in 7 batches of G. veitchiorum samples by HPLC-UV method. The method established in this study is simple and reliable, capable of qualitatively and quantitatively analyzing the main chemical components of G. veitchiorum, and is applicable to its quality evaluation.

Objective:To explore the impact of lymph node ratio (LNR) on the prognosis of gastric cancer patients.Methods:A retrospective case series study was conducted. The clinical data of 146 gastric cancer patients who underwent radical surgery and standardized chemotherapy at Lianyungang Department of General Surgery, Lianyungang Municipal Oriental Hospital from January 2015 to January 2019 were retrospectively analyzed. LNR was calculated, and receiver operating characteristic (ROC) curves of LNR were plotted based on overall survival (OS) to find the optimal cut-off value, and the patients were divided into high LNR group and low LNR group according to the cut-off value. The clinicopathological characteristics and prognosis of the two groups of patients were analyzed, and further the survival difference between patients aged ≥ 65 years and <65 years were analyzed in the two groups.Results:Out of 146 patients, 103 were male and 43 were female, with an age of (69±2) years; Pearson correlation analysis showed a positive correlation between the number of metastatic lymph nodes and LNR ( r = 0.899, P < 0.001). There was no significant correlation between the total number of cleared lymph nodes and LNR ( r = -0.069, P = 0.410). The 5-year cumulative OS rate for all patients was 30.1%, with a median OS time of 42.5 months. According to the 5-year OS, the ROC curve of LNR was plotted, and the area under the curve (AUC) of LNR was 0.699 (95% CI: 0.614-0.785, P < 0.001), with the optimal cut-off value of 0.355. LNR was related to age, tumor differentiation degree and TNM staging (all P < 0.05). The results of multivariate Cox regression analysis showed that LNR, age, TNM staging, and tumor differentiation degree were factors affecting the OS of patients. According to Kaplan-Meier analysis, the 5-year OS rate of patients with high LNR was lower than that of patients with low LNR (10.3% vs. 42.0%, χ2 = 69.89, P < 0.001). Among all patients, the 5-year OS rate of patients aged ≥ 65 years was lower than that of patients aged < 65 years (18.8% vs. 50.0%, χ2 = 15.77, P < 0.001). In the high LNR group, the 5-year OS rate of patients aged ≥ 65 years was lower than that of patients aged <65 years (2.3% vs. 33.3%, χ2 = 6.29, P = 0.012); in the low LNR group, the 5-year OS rate of patients aged ≥ 65 years was lower than that of patients aged < 65 years (32.3% vs. 55.9%, χ2 = 4.78, P = 0.029). Conclusions:LNR is an independent prognostic factor for gastric cancer patients, and gastric cancer patients with LNR > 0.355 have poor prognosis. With the same LNR, the prognosis of elderly patients is worse than that of young patients.

The transition of cancer cells from epithelial state to mesenchymal state awarded hepatocellular carcinoma (HCC) stem cell properties and induced tumorigenicity, drug resistance, and high recurrence rate. Reversing the mesenchymal state to epithelial state by inducing mesenchymal-epithelial remodeling could inhibit the progression of HCC. Using high-throughput screening, chrysin was selected from natural products to reverse epithelial-mesenchymal transition (EMT) by selectively increasing CDH1 expression. The target identification suggested chrysin exerted its anti-HCC effect through covalently and specifically binding threonine 205 (Thr205) of alpha-enolase (ENO1). For the first time, we revealed that ENO1 bound β-catenin mRNA, and recruited YTHDF2 to identify the m6A modified β-catenin in the 3'-UTR region to degrade β-catenin mRNA. Eventually, the CDH1 gene expression was improved through the regulation of β-catenin mRNA. ENO1/β-catenin mRNA interaction might be a promising target for cellular plasticity reprogramming. Moreover, chrysin could mediate mesenchymal‒epithelial remodeling through increasing degradation of β-catenin mRNA by promoting the binding of ENO1 and β-catenin mRNA. To the best of our knowledge, chrysin is the first reported small molecule inducing β-catenin mRNA degradation through binding to ENO1. The water-soluble derivative of chrysin may be a natural product-derived lead compound for circumventing metastasis, recurrence, and drug resistance of HCC by mediating mesenchymal‒epithelial remodeling.

Screening carbonyl reductases with the ability to catalyze the reduction of complex carbonyl compounds is of great significance for the biosynthesis of R-tolvaptan(R-TVP). In this study, the target carbonyl reductase in the crude enzyme extract of rabbit liver was separated, purified, and identified by ammonium sulfate precipitation, gel-filtration chromatography, ion exchange chromatography, affinity chromatography, and protein mass spectrometry. With the rabbit liver genome as the template, the gene encoding the carbonyl reductase rlsr5 was amplified by PCR and the recombinant strain was successfully constructed. After RLSR5 was purified by affinity chromatography, its enzymatic properties were characterized. The results indicated that the gene sequence of rlsr5 was 972 bp, encoding a protein with a molecular weight of 40 kDa. RLSR5 was a dimeric protein, and each monomer was composed of a (α/β)₈-barrel structure. RLSR5 could asymmetrically reduce 7-chloro-1-[2-methyl-4-[(2- methylbenzoyl)amino]benzoyl]-5-oxo-2,3,4,5-tetrahydro-1H-1-benzazepine (prochiral ketone, PK) to synthesize R-TVP. The specific activity of the enzyme was 36.64 U/mg, and the optical purity of the product was 99%. This enzyme showcased the optimal performance at pH 6.0 and 30 °C. It was independent of metal ions, with the activity enhanced by Mn²⁺. This study lays a foundation for the biosynthesis of tolvaptan of optical grade.
Animals ; Rabbits ; Alcohol Oxidoreductases/biosynthesis* ; Recombinant Proteins/metabolism* ; Escherichia coli/metabolism* ; Liver/enzymology*

Objective:To establish and validate a nomogram for predicting the probability of central lymph node metastasis (CLNM) by evaluating risk factors in clinical N 0(cN 0) stage patients with iisolated papillary thyroid carcinoma of isthmus (PTCI). Methods:Retrospectively analyzed the clinicopathological data of 112 PTCI patients who underwent surgical treatment at Lianyungang Municipal Oriental Hospital between January 2019 and December 2024. There were 35 males and 77 females, with an average age of (47.2±12.6) years, ranging from 28 to 71 years. Univariate and multivariate logistic regression analyses were performed to identify independent risk factors for CLNM. Nomogram method was used to build the transfer risk prediction model and verify it.Results:The study enrolled 112 patients, with 42 cases (37.5%) exhibiting CLNM. ROC curve analysis demonstrated that tumor diameter predicted CLNM with an AUC of 0.781 (95% CI: 0.694-0.868), identifying an optimal cutoff value of 0.9 cm (sensitivity: 0.762, specificity: 0.700). Multivariate logistic regression revealed that CLNM was independently associated with age, tumor diameter, capsular invasion, and Hashimoto′s thyroiditis (HT), all P<0.05. The logistic regression analysis results were used to construct the nomogram model of CLNM risk prediction. Hosmer lemeshow test resulted χ2=6.876, P=0.442. The calibration curves of the model group and the validation group were basically close, with an average absolute error of 0.065, P=0.099. ROC curve analysis showed that the best cut-off value of nomogram was 160 points, C-index was 0.849 (95% CI: 0.771-0.927), sensitivity was 0.786, specificity was 0.729. Decision curve analysis (DCA) based on net benefit and threshold probability shows that the nomogram can obtain net benefit in a large threshold probability range (0.13-1.00). Temporary recurrent laryngeal nerve injury occurred in 5 patients (4.5%). Temporary parathyroid injury occurred in 4 cases (3.6%), all of which were normal within one month. Conclusions:For patients with cN 0 stage isolated PTCI, CLNM is related to age ≤45 years, tumor diameter>0.9 cm, envelope invasion and HT. HT is a protective factor for CLNM. The ROC curve, calibration curve and DCA curve verified internally showed that the nomogram obtained had good discrimination ability, calibration ability and clinical applicability. When the total score ≥ 160 points indicates that the risk of CLNM is increased.

Objective:To evaluate the clinical efficacy of Sanren Decoction combined with triple therapy in the treatment of Hp-related chronic atrophic gastritis (CAG).Methods:A randomized controlled trial was conducted. A total of 118 patients with Hp-related CAG admitted to Department of Spleen and Stomach Diseases of Xi'an Affiliated Hospital of Shaanxi University of Chinese Medicine from February 2021 to February 2023 were selected as observation subjects, and were divided into two groups according to random number table method, with 59 cases in each group. The control group received triple therapy (amoxicillin+clarithromycin+omeprazole), whereas the observation group was combined with modified Sanren Decoction on the basis of the control group. Both groups were continuously treated for 10 days. TCM symptoms were scored before and after treatment. Updated Sydney System (UAS) was applied to evaluate the gastric mucosal lesions. ELISA was applied to detect serum levels of cluster of differentiation 47 (CD47), VEGF and pepsinogenⅠ/pepsinogen Ⅱ (PGⅠ/PGⅡ). The Hp infection status was evaluated by Urea Breath Test (UBT). In addition, the adverse reactions were observed and recorded during treatment.Results:After treatment, the scores of gastric fullness, stomachache, acid reflux and bitter taste in observation group were lower than those in the control group ( t=4.52, 3.46, 4.34, 4.83, P<0.01), and the scores of gastric mucosal inflammation, atrophy, intestinal metaplasia and dysplasia were also lower compared to control group ( t=3.68, 2.88, 3.73, 4.45, P<0.01 or P<0.05). After treatment, the levels of serum CD47 [(4.46±1.29) μg/L vs. (5.01±1.37) μg/L, t=2.25] and VEGF [(70.53±10.47) ng/L vs. (79.34±10.56) ng/L, t=4.55] in observation group were lower ( P<0.05 or P<0.01), while the level of PGⅠ/PGⅡ [(12.45±1.78) vs. (10.82±1.62), t=5.20] was higher compared with that of the control group ( P<0.01). After treatment, the negative conversion rate of Hp was 96.61% (57/59) in observation group and that in control group was 81.36% (48/59) ( χ2=7.00, P=0.008). During treatment, the incidence of adverse reactions was 23.73% (14/59) in observation group and 15.25% (9/59) in control group, revealing no statistical significance between the two groups ( χ2=1.35, P=0.245). Conclusion:Modified Sanren Decoction combined with triple therapy can improve gastric mucosal lesions, reduce serum CD47 and VEGF levels, and increase PGⅠ/PGⅡ ratio and Hp negative conversion rate in patients with Hp-related CAG, with good safety.

Objective:To investigate the correlation of combined detection of p16 and Rb with high-risk human papilloma virus (HR-HPV) infection in non-oropharyngeal squamous cell carcinoma (NOPSCC) of the head and neck.Methods:A total of 68 NOPSCC cases of the head and neck (23 cases of the nasal cavity and paranasal sinuses and 45 cases of larynx) with complete clinical and pathological data, diagnosed at the Beijing Tongren Hospital, Capital Medical University, Beijing, China from November 2013 to December 2023, were collected. The expression of p16 and Rb was detected using immunohistochemistry of the EnVision two-step method, while the HR-HPV mRNA expression was detected using in situ hybridization. The concordance, sensitivity, and specificity of p16 alone and the combined detection of p16 and Rb for detecting HR-HPV infection were analyzed.Results:Among the 68 patients with NOPSCC, 53 were male and 15 were female, with a median age of 63.5 (range, 57.3 to 66.8) years. 41 patients had a smoking history and 27 did not. 33 patients had an early T stage (T1/T2) and 35 had advanced T stage (T3/T4). 14 patients had lymph node metastasis and 2 had distant metastasis. Histological types included 62 cases of keratinized squamous cell carcinoma, 5 cases of non-keratinized squamous cell carcinoma, and 1 case of basal-like squamous cell carcinoma. 25 cases were positive for p16. Among the 25 cases, 16 cases were positive for Rb, and 6 cases were positive for HR-HPV mRNA. 43 cases were negative for p16, including 38 cases positive for Rb and no cases positive for HR-HPV mRNA. The concordance between p16 and HR-HPV mRNA expression was poor ( Kappa=0.285, P=0.001), with a sensitivity of 100.0% and specificity of 69.4%. In contrast, the combined detection of p16+/Rb- showed high concordance with HR-HPV mRNA expression ( Kappa=0.719, P<0.001), with a sensitivity of 100.0% and specificity of 95.2%. Conclusions:In NOPSCC of the head and neck, the combined detection of p16 and Rb may be used as a marker for assessing HR-HPV infection. Recognizing the p16+/Rb- expression pattern in NOPSCC can improve the specificity of HR-HPV detection.

Objective:To investigate the correlation of combined detection of p16 and Rb with high-risk human papilloma virus (HR-HPV) infection in non-oropharyngeal squamous cell carcinoma (NOPSCC) of the head and neck.Methods:A total of 68 NOPSCC cases of the head and neck (23 cases of the nasal cavity and paranasal sinuses and 45 cases of larynx) with complete clinical and pathological data, diagnosed at the Beijing Tongren Hospital, Capital Medical University, Beijing, China from November 2013 to December 2023, were collected. The expression of p16 and Rb was detected using immunohistochemistry of the EnVision two-step method, while the HR-HPV mRNA expression was detected using in situ hybridization. The concordance, sensitivity, and specificity of p16 alone and the combined detection of p16 and Rb for detecting HR-HPV infection were analyzed.Results:Among the 68 patients with NOPSCC, 53 were male and 15 were female, with a median age of 63.5 (range, 57.3 to 66.8) years. 41 patients had a smoking history and 27 did not. 33 patients had an early T stage (T1/T2) and 35 had advanced T stage (T3/T4). 14 patients had lymph node metastasis and 2 had distant metastasis. Histological types included 62 cases of keratinized squamous cell carcinoma, 5 cases of non-keratinized squamous cell carcinoma, and 1 case of basal-like squamous cell carcinoma. 25 cases were positive for p16. Among the 25 cases, 16 cases were positive for Rb, and 6 cases were positive for HR-HPV mRNA. 43 cases were negative for p16, including 38 cases positive for Rb and no cases positive for HR-HPV mRNA. The concordance between p16 and HR-HPV mRNA expression was poor ( Kappa=0.285, P=0.001), with a sensitivity of 100.0% and specificity of 69.4%. In contrast, the combined detection of p16+/Rb- showed high concordance with HR-HPV mRNA expression ( Kappa=0.719, P<0.001), with a sensitivity of 100.0% and specificity of 95.2%. Conclusions:In NOPSCC of the head and neck, the combined detection of p16 and Rb may be used as a marker for assessing HR-HPV infection. Recognizing the p16+/Rb- expression pattern in NOPSCC can improve the specificity of HR-HPV detection.

Objective:To explore the impact of lymph node ratio (LNR) on the prognosis of gastric cancer patients.Methods:A retrospective case series study was conducted. The clinical data of 146 gastric cancer patients who underwent radical surgery and standardized chemotherapy at Lianyungang Department of General Surgery, Lianyungang Municipal Oriental Hospital from January 2015 to January 2019 were retrospectively analyzed. LNR was calculated, and receiver operating characteristic (ROC) curves of LNR were plotted based on overall survival (OS) to find the optimal cut-off value, and the patients were divided into high LNR group and low LNR group according to the cut-off value. The clinicopathological characteristics and prognosis of the two groups of patients were analyzed, and further the survival difference between patients aged ≥ 65 years and <65 years were analyzed in the two groups.Results:Out of 146 patients, 103 were male and 43 were female, with an age of (69±2) years; Pearson correlation analysis showed a positive correlation between the number of metastatic lymph nodes and LNR ( r = 0.899, P < 0.001). There was no significant correlation between the total number of cleared lymph nodes and LNR ( r = -0.069, P = 0.410). The 5-year cumulative OS rate for all patients was 30.1%, with a median OS time of 42.5 months. According to the 5-year OS, the ROC curve of LNR was plotted, and the area under the curve (AUC) of LNR was 0.699 (95% CI: 0.614-0.785, P < 0.001), with the optimal cut-off value of 0.355. LNR was related to age, tumor differentiation degree and TNM staging (all P < 0.05). The results of multivariate Cox regression analysis showed that LNR, age, TNM staging, and tumor differentiation degree were factors affecting the OS of patients. According to Kaplan-Meier analysis, the 5-year OS rate of patients with high LNR was lower than that of patients with low LNR (10.3% vs. 42.0%, χ2 = 69.89, P < 0.001). Among all patients, the 5-year OS rate of patients aged ≥ 65 years was lower than that of patients aged < 65 years (18.8% vs. 50.0%, χ2 = 15.77, P < 0.001). In the high LNR group, the 5-year OS rate of patients aged ≥ 65 years was lower than that of patients aged <65 years (2.3% vs. 33.3%, χ2 = 6.29, P = 0.012); in the low LNR group, the 5-year OS rate of patients aged ≥ 65 years was lower than that of patients aged < 65 years (32.3% vs. 55.9%, χ2 = 4.78, P = 0.029). Conclusions:LNR is an independent prognostic factor for gastric cancer patients, and gastric cancer patients with LNR > 0.355 have poor prognosis. With the same LNR, the prognosis of elderly patients is worse than that of young patients.

In order to understand the current status of the knowledge, attitude and practice (KAP) about prevention and control of COVID -19 in college students, and to provide theoretical basis for prevention and control work in college campus. This study investigated the KAP of COVID -19 of 1 847 college students in Shaanxi province by questionnaire using the convenience sampling method. Chisquare test and multivariate logistic regression analysis were used to analyze the influencing factors for the KAP of COVID -19. The results demonstrated that 48.3% of the students had a higher knowledge level of COVID -19, 11.7% had a fear attitude and 39.6% had good protective practices. Logistic regression results showed that female and urban household college students had higher cognitive level of COVID -19. The college students with anxiety state were more likely to have fear attitude. Students of female, urban household, anxiety, higher cognition and fear attitude showed better protective practices. The above results indicated that the knowledge level of COVID -19 in college students are not enough, and the attitude and protective practices need to be further improved. Therefore, relevant departments should follow the rules of KAP, carry out targeted propaganda and education on COVID -19 for college students, to improve their ability to cope with public health emergencies.