Objective To establish an HPLC method to determine the concentration of inositol nicotinate（IN） in rat skin, and study the pharmacokinetic characteristics of IN after transdermal administration of heparin sodium inositol nicotinate cream in rats. Methods HPLC method was used to establish a simple and rapid analytical method for the determination of IN concentration in the skin of rats at different time points after administration. The established method was used to study the pharmacokinetics of IN after transdermal administration of heparin sodium inositol nicotinate cream in rats, and the pharmacokinetic parameters were fitted with DAS software. Results The linearity of the analytical method was good in the concentration range of 0.25-20 μg/ml, the quantitative limit was 0.25 μg/ml, and the average recovery rate was 96.18%. The pharmacokinetic parameters of IN after transdermal administration of heparin sodium inositol nicotinate cream in rats were as follows: t_1/2 was （4.555±2.054） h, T_max was （6±0）h, C_max was （16.929±2.153）mg/L, AUC_0−t was （150.665±16.568） mg·h /L ,AUC_0−∞ was （161.074±23.917） mg·h /L, MRT_（0−t） was （9.044±0.618）h, MRT_{（0−∞）} was （10.444±1.91） h, CLz/F was （0.19±0.03） L/（h·kg）, and Vz/F was （1.19±0.437） L/（h·kg）. Conclusion IN could quickly penetrate the skin and accumulate in the skin for a long time, which was beneficial to the pharmacological action of drugs on the lesion site for a long time. The method is simple, rapid, specific and reproducible, which could be successfully applied to the pharmacokinetic study of IN after transdermal administration in rats.

Objective To investigatethe relationship between gastroesophageal reflux disease (GERD) symptoms and clinicopathological characteristics, p53 expression, and survival of Chinese patients with esophageal adenocarcinoma. Methods A total of 3882 patients with esophageal adenocarcinoma, 970 matched patients with esophageal squamous cell carcinoma, and 970 matched patients with gastric cardia adenocarcinoma were included to compare the incidence of GERD symptoms. Patients with esophageal adenocarcinoma were divided into two groups according to the presence and absence of GERD symptoms. The differences in clinicopathological characteristics and p53 expression between the two groups were compared by chi-square test, and the differences in survival between the two groups were compared by landmark analysis. Results The incidence of GERD symptoms increased successively in esophageal squamous cell carcinoma, esophageal adenocarcinoma, and gastric cardia adenocarcinoma. In patients with esophageal adenocarcinoma, the proportions of male, less than 65 years old, lower thoracic tumors, tumors without squamous cell carcinoma, poorly differentiation, early tumors (stage 0-I), and p53-positive tumors in the group with GERD symptoms were higher than those in the group without GERD symptoms; moreover, the long-term survival (≥15 years) was better. Conclusion GERD symptom is an important clinical sign of esophageal adenocarcinoma. It is closely related to specific clinicopathological characteristics, p53 overexpression, and good long-term prognosis.

Objective To establish the fingerprint of Jianggui granules, and evaluate it by chemometrics. Methods The fingerprint of Jianggui granules was established by HPLC. Similarity evaluation system of chromatographic fingerprint of TCM （2012 edition） was used to evaluate the similarity evaluation. Then, the quality of the drug was assessed by cluster analysis （CA）, principal components analysis （PCA） and partial least squares discriminant analysis （PLS-DA）. Results The characteristic fingerprint of Jianggui granules was established and 18 common peaks were verified. Five chromatographic peaks were identified，i.e. Puerarin, glycyrrhizin, cinnamic acid, cinnamaldehyde and ammonium glycyrrhizinate. The similarities of samples were >0.9. Results of CA showed that 14 batches of samples could be classified into two categories：S1 and S4 were grouped into one category；others were grouped into the other category. The results of PCA showed that the cumulative contribution rate of the first two principal components was 96.61%. The results of OPLS-DA showed that the eleven peaks with VIP value >1 were puerarin （peak 8）, glycyrrhizin （peak 14）, cinnamaldehyde （peak 17） and ammonium glycyrrhizinate （peak 18）. Conclusion HPLC fingerprint of Jianggui granules was established. The established method was accurate and reliable，which could be used in quality evaluation of Jianggui granules.

This study aimed to explore the therapeutic mechanisms of Colquhounia Root Tablets(CRT) in treating diabetic kidney disease(DKD) by integrating biomolecular network mining with animal model verification. By analyzing clinical transcriptomics data, an interaction network was constructed between candidate targets of CRT and DKD-related genes. Based on the topological eigenvalues of network nodes, 101 core network targets of CRT against DKD were identified. These targets were found to be closely related to multiple pathways associated with type 2 diabetes, immune response, and metabolic reprogramming. Given that immune-inflammatory imbalance driven by metabolic reprogramming is one of the key pathogenic mechanisms of DKD, and that many core network targets of CRT are involved in this pathological process, receptor for advanced glycation end products(RAGE)-reactive oxygen species(ROS)-phosphatidylinositol 3-kinase(PI3K)-protein kinase B(AKT)-nuclear factor-κB(NF-κB)-NOD-like receptor family pyrin domain containing 3(NLRP3) signaling axis was selected as a candidate target for in-depth research. Further, a rat model of DKD induced by a high-sugar, high-fat diet and streptozotocin was established to evaluate the pharmacological effects of CRT and verify the expression of related targets. The experimental results showed that CRT could effectively correct metabolic disturbances in DKD, restore immune-inflammatory balance, and improve renal function and its pathological changes by inhibiting the activation of the RAGE-ROS-PI3K-AKT-NF-κB-NLRP3 signaling axis. In conclusion, this study reveals that CRT alleviates the progression of DKD through dual regulation of metabolic reprogramming and immune-inflammatory responses, providing strong experimental evidence for its clinical application in DKD.
Animals ; Diabetic Nephropathies/metabolism* ; Receptor for Advanced Glycation End Products/genetics* ; NF-kappa B/genetics* ; Signal Transduction/drug effects* ; Rats ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Proto-Oncogene Proteins c-akt/genetics* ; Drugs, Chinese Herbal/administration & dosage* ; Male ; Phosphatidylinositol 3-Kinases/genetics* ; Reactive Oxygen Species/metabolism* ; Humans ; Plant Roots/chemistry* ; Rats, Sprague-Dawley ; Tablets/administration & dosage*

Objective:To investigate the related factors affecting the survival of adult patients with newly diagnosed chronic myeloid leukemia in the chronic phase (CML-CP) treated with tyrosine kinase inhibitor (TKI) and the prognostic predictive effect of the nomogram constructed according to them.Methods:A retrospective cohort study was conducted. Clinical general information and laboratory index data of 243 newly diagnosed adult CML-CP patients treated with TKI in the First Hospital of Lanzhou University from December 2008 to June 2023 were collected, and they were divided into a training set (194 patients) and a validation set (49 patients) by complete randomization in the ratio of 8∶2. In the training set, variables affecting poor overall survival (OS) of patients were analyzed using univariate and multivariate Cox proportional hazards models by R4.3.2 software to obtain the independent influences of poor OS, on the basis of which the Cox regression model was constructed and the nomogram predicting the OS rate at 8 and 10 years was plotted. Kaplan-Meier method was applied to analyze the OS in all 243 patients and patients stratified by the screened independent influencing factors for poor OS, and log-rank test was used for comparison between the groups. In the training and validation sets, receiver operating characteristic (ROC) curve was used to analyze the effect of the nomogram on predicting 8- and 10-year OS rates of patients with actual survival as the gold standard; calibration curve was used to assess the accuracy of predictions of the nomogram; decision curve analysis (DCA) was used to assess the clinical utility of the nomogram.Results:The median age of 243 CML-CP patients [ M ( Q1, Q3)] was 46 (35, 58) years old, 150 cases (61.7%) were male and 9 cases (38.3%) were female, 119 cases (49.0%) had comorbidities, and the efficacy of 82 cases (33.7%) reached molecular response (MR) 5.0. Differences in patient compositions for age and gender, levels of major indicators for peripheral blood and bone marrow, spleen size, comorbidities, short-term efficacy, Sokal score, and long-term survival score of European Treatment and Outcomes Study between the training and validation sets were not statistically significant (all P > 0.05). Multivariate Cox regression analysis showed that patients with elevated age ( HR = 1.04, 95% CI: 1.01-1.08, P = 0.041), comorbidities (with vs. without, HR = 3.48, 95% CI: 1.23-9.86, P = 0.019), and those who did not achieve MR5.0 (achieved vs. unachieved, HR = 0.13, 95% CI: 0.02-0.97, P = 0.046) were independent risk factors for poor OS in TKI-treated newly diagnosed adult CML-CP patients. By the last follow-up (December 2023), the median follow-up was 72 months, with the range of 6-180 months. Kaplan-Meier method analysis showed that the 8- and 10-year OS rates of 243 patients were 83.7% and 81.6%, respectively; patients with age ≥46 years compared to <46 years, with comorbidities compared to without comorbidities, and who did not achieve MR5.0 in terms of efficacy compared to who achieved MR5.0, the OS was poorer, and the differences were statistically significant (all P < 0.01). The nomogram of 8- and 10-year OS rates in TKI-treated newly diagnosed adult CML-CP patients was constructed based on the screened independent influencing factors of poor OS. The area under the ROC curve was 0.910 and 0.851 in the training set and 0.778 and 0.764 in the validation set for the predicted 8- and 10-year OS rates based on the nomogram, respectively, and the calibration curve showed that the predicted 8- and 10-year OS rates based on the nomogram were in high agreement with the actual ones in the training and validation sets; the DCA showed that the nomogram within a certain prediction threshold could benefit the clinical decision-making in both the training and validation sets. Conclusions:Having comorbidities, not reaching MR5.0 in efficacy and old age are independent risk factors for poor survival of TKI-treated adult patients with newly diagnosed CML-CP, and the nomogram constructed based on these 3 factors has a good predictive ability for the survival of such patients.

Objective To develop and validate risk prediction models utilizing five machine learning algorithms for assessing postoperative pulmonary infection(PPI)risk in lung cancer patients undergoing grade Ⅳ thoracoscopic surgery.Methods A retrospective cohort study included 2,380 lung cancer patients who underwent grade Ⅳ thoracoscopic surgery at a tertiary hospital in Shanghai(January 2022 to June 2024).Patients were stratified into training(n=1,665)and validation(n=715)cohorts.Five machine learning algorithms—Logistic regression(LR),artificial neural network(ANN),support vector machine(S VM),random forest(RF),and extreme gradient boosting(XGB)—were employed to construct predictive models.A nomogram was developed for clinical utility.Results Among 2,380 patients,226(9.5％)developed PPI.The Least Absolute Shrinkage and Se-lection Operator(LASSO)regression identified eight predictive variables:daily cigarette consumption,diabetes history,preoperative diffusing capacity,maximal tumor diameter,24-hour postoperative chest drainage volume,perioperative oral nutritional supplementation(ONS),postoperative urinary cathe-terization,and intraoperative pleural adhesion severity.All models demonstrated robust discrimina-tion,with area under the curve(AUC)values ranging from 0.862 to 0.947.The XGB model a-chieved superior performance(AUC=0.947,95％CI,0.937 to 0.962),followed closely by the LR model(AUC=0.926,95％CI,0.918 to 0.933).Conclusion Machine learning-based algo-rithms models effectively stratify PPI risk in lung cancer patients following grade Ⅳ thoracoscopic surgery.The derived nomogram provides a practical tool for perioperative risk management by healthcare providers.

Objective To explore the symptom clusters in renal transplant recipients and con-struct a concurrent symptom network to identify core symptoms.Methods A total of 343 patients with followed up after renal transplantation were selected as the study subjects.A general information questionnaire and the Chinese version of the Modified Transplant Symptom Occurrence and Symptom Distress Scale were employed to analyze the occurrence of symptoms in patients.In this study,only symptoms with an incidence rate greater than 20％and Spearman correlation coefficient greater than 0.40 between symptom severity and total score were retained.Exploratory factor analysis was used to extract symptom clusters with a factor loading of ≥0.45 as the criterion.The R language was utilized to construct symptom network,based on which core symptoms and bridge symptoms were identified.Results A total of 5 symptom clusters were extracted in this study:the neuro-gastrointestinal symp-tom cluster,the mood-related symptom cluster,the hormone-related symptom cluster,the energy de-ficiency symptom cluster and the vision-related symptom cluster.The core symptoms were anxiety(rs=1.75),mood swings(rs=1.50)and muscle weakness(rs=1.27).The top three symptoms in terms of bridge strength were muscle weakness(rb=0.87),lack of vitality(rb=0.66)and fa-tigue(rb=0.65).Conclusion Multiple symptoms are presented in patients after renal transplanta-tion.Based on the results of symptom network analysis,clinicians can strengthen the assessment of core symptoms and bridge symptoms to develop precise intervention strategies and improve the effectiveness of symptom management.

Objective To explore the effect of intestinal nitrates on the growth of Klebsiella pneumoniae and its regulatory mechanisms. Methods K. pneumoniae strains with nitrate reductase narG and narZ single or double gene knockout or with NarXL gene knockout were constructed and observed for both aerobic and anaerobic growth in the presence of KNO3 using an automated bacterial growth analyzer and a spectrophotometer, respectively. The mRNA expressions of narG and narZ in K. pneumoniae in anaerobic cultures in the presence of KNO3 and the effect of the binary regulatory system NarXL on their expresisons were detected using qRT-PCR. Electrophoretic mobility shift assays (EMSA) and MST analysis were performed to explore the specific regulatory mechanisms of NarXL in sensing and utilizing nitrates. Competitive experiments were conducted to examine anaerobic growth advantages of narG and narZ gene knockout strains of K. pneumoniae in the presence of KNO3. Results The presence of KNO3 in anaerobic conditions, but not in aerobic conditions, promoted bacterial growth more effectively in the wild-type K. pneumoniae strain than in the narXL gene knockout strain. In anaerobic conditions, the narXL gene knockout strain showed significantly lowered mRNA expressions of narG and narZ (P<0.0001). EMSA and MST experiments demonstrated that the NarXL regulator could directly bind to narG and narZ promoter regions. The wild-type K. pneumoniae strain in anaerobic cultures showed significantly increased expressions of narG and narZ mRNAs in the presence of KNO3 (P<0.01), and narG gene knockout resulted in significantly attenuated anaerobic growth and competitive growth abilities of K. pneumoniae in the presence of KNO3 (P<0.01). Conclusion The binary regulatory system NarXL of K. pneumoniae can sense changes in intestinal nitrate concentration and directly regulate the expression of nitrate reductase genes narG and narZ to promote bacterial growth.

Objective:To explore the cause of inconsistency between the results of trisomy 7 by expanded non-invasive prenatal testing (NIPT-PLUS) and trisomy 18 by prenatal diagnosis.Methods:A pregnant woman who received genetic counseling at Jiaozuo Maternal and Child Health Care Hospital on July 5, 2020 was selected as the study subject. NIPT-PLUS, systematic ultrasound and interventional prenatal testing were carried out. The middle segment and root of umbilical cord, center and edge of the maternal and fatal surface of the placenta were sampled for the validation by copy number variation sequencing (CNV-seq).Results:The result of NIPT-PLUS indicated that the fetus has trisomy 7. Systematic ultrasound has shown multiple malformations including atrioventricular septal defect, horseshoe kidney, and rocker-bottom feet. However, QF-PCR, chromosomal karyotyping analysis, and CNV-seq of amniotic fluid samples all showed that the fetus was trisomy 18. Validation using multiple placental samples confirmed that the middle segment of the umbilical cord contains trisomy 18, the center of the placenta contained trisomy 7, and other placental sites were mosaicism for trisomy 7 and trisomy 18. Notably, the ratio of trisomy 18 became lower further away from the umbilical cord.Conclusion:The false positive results of trisomy 7 and false negative trisomy 18 by NIPT-PLUS was probably due to the existence of placental mosaicism. Strict prenatal diagnosis is required needed aneuploidy is detected by NIPT-PLUS to exclude the influence of placental mosaicisms.

This study design of specific identification primers for the ITS2 sequence of F. ussuriensis. The reaction system and conditions were optimized, and PCR-nucleic acid test strips were constructed to realize the visual detection of F. ussuriensis Bark. Through molecular cloning and sequencing technology, we constructed a positive control for F. ussuriensis DNA and formulated quality standards. The established method was evaluated for sensitivity, specificity and reproducibility, and the authenticity of the commercially available samples was identified. Results demonstrated that based on the ITS2 sequences, F. ussuriensis and its mixed forgeries could be distinguished. The PCR products of the authentic F. ussuriensis on test strips showed two bands in the T and C lines, while the pseudo products and negative control showed only one band in the C line, which was consistent with the results of agarose gel electrophoresis. The specificity was 100%, and the sensitivity of the PCR-nucleic acid test strip was up to 0.1 ng·μL^-1, which was 10 times higher than that of the gel electrophoresis assay. 11 out of 16 commercially available samples of F. ussuriensis were qualified, and 5 were unqualified. Collectively, the PCR-nucleic acid test strip method established in this study is specific, rapid, accurate and visualized, which can provide a new technical idea for the detection of F. ussuriensis.