Objective Based on the Ancient and Modern Medical Case Cloud Platform V2.3.7,analyze the medication pattern and mechanism of action of Chinese medicines commonly used in famous medical cases for the treatment of Alzheimer's disease(AD).Methods The core Chinese medicines for AD were screened according to the inclusion and exclusion criteria by searching the famous Chinese medicine cases in the platform,then established the disease-core Chinese medicines-active ingredients-common targets network.The protein-protein interaction network(PPI)was established through the common targets.Gene oncology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed by David platform.Molecular docking and dynamics simulations were employed to evaluate the binding affinity and stability between key targets and active ingredients.Results Data mining was performed on all the famous medical cases from the time of database construction to October 2023,and the results were summarized as follows.The high-frequency regulating Qi medications and tonifying Qi medications were Citri Reticulatae Pericarpium and Aurantii Fructus Immaturus;Panax ginseng,Ziziphi Jujubae Fructus,Codonopsis pilosula,Astragali Radix,Atractylodis Macrocephalae Rhizoma,Dioscoreae Rhizoma,and Glycyrrhizae Radix et Rhizoma.The core herbs contained 778 active ingredients and 377 targets;1986 disease targets;79 common targets were obtained after intersection with AD targets,and 7 core targets were identified through PPI network topology analysis,including TNF,AKT1,TP53,PPARG,etc.GO and KEGG pathway enrichment analyses confirmed that the targets of the active ingredients of the core herbs involved 138 pathways,and the core herbs could regulate chemical carcinogenesis-receptor activation、serotonergic synapse、chemical carcinogenesis-reactive oxygen species pathways to treat AD.Molecular docking and molecular dynamics simulation results showed good binding ability and stability between the key targets and the core components.Conclusion Through data mining to analyze the core herbs for the treatment of AD,the present investigation showing the pharmacological mechanism of 12-O-Nicotinoylisolineolone and Odoratin in the treatment of Alzheimer's disease at the molecular level,lay a certain theoretical foundation of 12-O-Nicotinoylisolineolone and Odoratin in the future.

Objective Based on the Ancient and Modern Medical Case Cloud Platform V2.3.7,analyze the medication pattern and mechanism of action of Chinese medicines commonly used in famous medical cases for the treatment of Alzheimer's disease(AD).Methods The core Chinese medicines for AD were screened according to the inclusion and exclusion criteria by searching the famous Chinese medicine cases in the platform,then established the disease-core Chinese medicines-active ingredients-common targets network.The protein-protein interaction network(PPI)was established through the common targets.Gene oncology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed by David platform.Molecular docking and dynamics simulations were employed to evaluate the binding affinity and stability between key targets and active ingredients.Results Data mining was performed on all the famous medical cases from the time of database construction to October 2023,and the results were summarized as follows.The high-frequency regulating Qi medications and tonifying Qi medications were Citri Reticulatae Pericarpium and Aurantii Fructus Immaturus;Panax ginseng,Ziziphi Jujubae Fructus,Codonopsis pilosula,Astragali Radix,Atractylodis Macrocephalae Rhizoma,Dioscoreae Rhizoma,and Glycyrrhizae Radix et Rhizoma.The core herbs contained 778 active ingredients and 377 targets;1986 disease targets;79 common targets were obtained after intersection with AD targets,and 7 core targets were identified through PPI network topology analysis,including TNF,AKT1,TP53,PPARG,etc.GO and KEGG pathway enrichment analyses confirmed that the targets of the active ingredients of the core herbs involved 138 pathways,and the core herbs could regulate chemical carcinogenesis-receptor activation、serotonergic synapse、chemical carcinogenesis-reactive oxygen species pathways to treat AD.Molecular docking and molecular dynamics simulation results showed good binding ability and stability between the key targets and the core components.Conclusion Through data mining to analyze the core herbs for the treatment of AD,the present investigation showing the pharmacological mechanism of 12-O-Nicotinoylisolineolone and Odoratin in the treatment of Alzheimer's disease at the molecular level,lay a certain theoretical foundation of 12-O-Nicotinoylisolineolone and Odoratin in the future.

"Quality evaluation through identification"(QECI)is a theory that summarizes the essence of quality identification experience about traditional Chinese medicine(TCM).As an important component of the current quality evaluation system of TCM,the theory provides theoretical guidance for controlling and evaluating TCM's quality.Identifying based on human senses,traditional appearance identification relies on experience,which is subjective,difficult to quantify,and poorly repeatable.Modern measurement tools and biomimetic technology can quantify appearance characters(shape,color,smell,taste)of TCM.Then associate the results with content of active ingredients or biological effects.Make use of the visualization,quantifiability,portability,and repeatability of modern scientific means and screen feasible and controllable quality control indicators,which can be applied to quality evaluation and control of TCM's production and processing in each stage.The origin,connotation,modern research progress,and application value of QECI are reviewed,aiming to provide reference for addressing the shortcomings of traditional appearance evaluation,expanding the application scope of QECI,and promoting the construction of TCM's quality evaluation system.

ObjectiveTo investigate the value of interleukin-32 (IL-32) combined with Model for End-Stage Liver Disease (MELD) in predicting the prognosis of patients with hepatitis B virus (HBV)-related acute-on-chronic liver failure (HBV-ACLF). MethodsA total of 92 patients with HBV-ACLF who were hospitalized in The First Affiliated Hospital of Soochow University from January 2015 to December 2018 were enrolled, and according to the follow-up results at 3 months after diagnosis, the patients were divided into survival group with 40 patients and death group with 52 patients. ELISA was used to measure the serum level of IL-32. Clinical data of the patients were collected, including age, sex, underlying diseases, major complications, white blood cell count (WBC), platelet count (PLT), hematocrit (HCT), total bilirubin (TBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (Alb), serum creatinine (SCr), prothrombin time (PT), international normalized ratio (INR), and HBV DNA. The t-test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups; a Pearson correlation analysis was performed for IL-32 and other variables; a binary logistic regression analysis was performed to investigate the independent risk factors for the prognosis of patients with HBV-ACLF. The receiver operating characteristic(ROC) curve(AUC) was used to evaluate the value of IL-32 combined with MELD score in predicting the prognosis of patients with HBV-ACLF. The normal Z test was used for comparison of AUC. ResultsThere were significant differences between the two groups in HCT, PLT, TBil, SCr, PT, INR, HBV DNA, IL-32, and MELD score (all P＜0.05). IL-32 was positively correlated with TBil (r=0.952, P＜0.001) and MELD score (r=0850, P＜0.001). IL-32 (odds ratio ［OR］=1.137, 95% confidence interval ［CI］: 1040-1243, P=0.005) and MELD score (OR=1.055, 95% CI: 1.001-1.109, P=0.025) were independent risk factors for the death of HBV-ACLF patients. IL-32 combined with MELD score had the highest value in predicting the prognosis of patients with HBV-ACLF (AUC=0.992, 95% CI: 0.981-1000), with a significantly higher AUC than IL-32 (0.992 vs 0.984, Z=2.265, P＜0.05) and MELD score (0.992 vs 0877, Z=3182, P＜0.05). ConclusionBoth IL-32 and MELD score can predict the prognosis of patients with HBV-ACLF, and the combination of these two indicators has a better predictive value.

ObjectiveTo investigate the level of vascular endothelial growth factor (VEGF) in the peripheral blood of patients with liver cirrhosis and portal vein thrombosis (PVT) and its clinical significance in the diagnosis of liver cirrhosis with PVT. MethodsA total of 60 patients with liver cirrhosis and PVT who were followed up or hospitalized in the outpatient service of The First Affiliated Hospital of Soochow University from January 2017 to December 2019 were enrolled as PVT group, and 161 patients with liver cirrhosis and portal hypertension who had no thrombi were enrolled as LC group. Related clinical data were collected, including sex, age, white blood cell count, platelet count, total bilirubin, alanine aminotransferase, albumin, prothrombin time, international normalized ratio, and antithrombin Ⅲ. Double-antibody sandwich ELISA was used to measure the serum level of VEGF. The t-test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. The Pearson correlation test was used for correlation analysis, and a binary logistic regression analysis was used to investigate the risk factors for PVT in patients with liver cirrhosis. ResultsThe comparison of baseline indices showed that there was a significant difference in serum VEGF between the PVT group and the LC group (P＜0.05). The comparison of the patients with Child-Pugh class A, B or C disease showed that there was a significant difference in serum VEGF between the two groups (Z=3.749， 5.469， all P＜0.05). The stepwise logistic regression analysis showed that only serum VEGF (odds ratio=1004, 95% confidence interval: 1.003-1.006, P＜0.001) was an independent risk factor for liver cirrhosis with PVT. ConclusionPatients with liver cirrhosis and PVT tend to have a high level of VEGF in peripheral blood, which provides guidance for clinical diagnosis.

Objective: To analyze the subjective and objective misdiagnostic reasons of brucellosis in Suzhou City. Methods: The clinical data of inpatients with brucellosis from the department of infectious diseases, the First Affiliated Hospital of Soochow University from 2016 to 2018 were collected. The main onset methods, the first consultation department, the diagnosis and treatment process, and the main diagnosis were analyzed. Simultaneously, a questionnaire survey was conducted among doctors from different departments in Suzhou City. To investigate the knowledge and diagnosis of brucellosis by electronic questionnaire, and analyze the results. Results: Totally 24 brucellosis cases were collected. Fever (13 cases, 54.17%), lumbago/arthralgia (6 cases, 25.00%) or both (3 cases, 12.50%) were the main initial symptoms, while a patient with fever, arthralgia and orchitis(1 case, 4.17%) and another with purpura (1 case, 4.17%) were also admitted. The 24 patients were distributed among 10 different departments, in which the department of infectious diseases (6 cases, 25.00%) and orthopedics (5 cases, 20.83%) dominated. Brucellosis was considered in only 2 patients (8.33%) before pathogen results, and the main suspected diagnosis were malignant hematopathy/lymphoma (7 cases, 29.17%), sepsis (5 cases, 20.83%), and tuberculosis (4 cases, 16.67%). Totally 124 questionnaires were collected and analyzed. We divided them into 2 groups: department of infectious diseases (52/124, 41.94%) and other departments (72/124, 58.06%). The proportion of answer "yes" for the question Suzhou is the epidemic area of brucellosis, question whether brucellosis is considered in suspected patients, question whether the epidemiology is collected and question how and where to conduct the serological test was 38.71% (48/124), 32.26% (40/124), 53.23% (66/124) and 29.84% (37/124), respectively. The positive answer in the department of infectious diseases (24/52, 46.15%) was significantly higher than those of others (16/72, 22.22%, χ²=7.913, P < 0.05). The respondents thought the serological test could not be conducted, because of that the non-pasture doctors were not familiar with the test procedure, which was complex (93/124, 75.00%) and the disease was not considered (85/124, 68.55%). And the respondents thought measures helpful to reduce misdiagnosis should include setting up serological test or simplifying the procedure (96/124, 77.42%), massed learning organized by related administrations (95/124, 76.61%) and learning by books, papers and classes (72/124, 58.06%). Conclusion There is obvious clinical misdiagnosis of brucellosis in Suzhou City, which may be caused by Suzhou doctors not paying enough attention to brucellosis, being unfamiliar with the inspection process and complicated inspection process.

Objective: To explore the prognostic value of model for end-stage liver disease (MELD) combined with arterial blood lactate (Lac) in admitted patients with hepatitis B virus-associated acute- on-chronic liver failure (HBV-ACLF). Methods: Clinical data of 97 cases with hepatitis B virus-associated acute- on-chronic liver failure (HBV-ACLF) admitted to the First Affiliated Hospital of Suzhou University between March 2016 and March 2018 was retrospectively analyzed. Age, gender, complications, MELD score, lactic acid (Lac), total bilirubin (TBil), creatinine (Cr), serum albumin (Alb), high-sensitivity C-reactive protein (CRP), white blood cell count (WBC), platelet count (PLT), hematocrit (Hct), quantification of HBV DNA and HBsAg, and organ support treatment (artificial liver support system, renal replacement therapy and mechanical ventilation ) were documented after admission. The primary endpoint of treatment was death due to ineffective medical treatment during hospitalization, abandonment of medical treatment due to deterioration of the health condition, and switch to liver transplantation for patients with poor medical treatment. The risk factors for primary endpoint of treatment were analyzed by binary logistic regression. Hosmer-Lemeshow test was used to evaluate the goodness of fit for the scoring system, and the ROC to predict the prognosis of MELD-Lac. Results: Ninety-seven cases with HBV-ACLF were included, 56 cases had good prognosis, and 41 cases had bad prognosis (including two cases with poor medical treatment and liver transplantation). The overall improvement rate was 57.7%. MELD score and Lac value in treated group was significantly lower than non-treated group. Bivariable and multivariable logistic regression analysis showed that the MELD score [odds ratio (OR = 1.806)], and Lac score [odds ratio (OR = 1.820)] was the risk factor for hospitalization and mortality in patients with liver failure (P < 0.05). The area under the ROC curve (AUC) and the 95% confidence interval (95% CI) of prognostic patients with MELD-Lac were significantly better than Lac and MELD scores [0.923 (0.84 to 1.00) vs. 0.804 (0.067 to 0.942) and 0.864 (0.75). 0.977)], P < 0.05. When the MELD-Lac Youden index was set at 0.746, the optimal threshold was 18.36, and the sensitivity and specificity were 91.3% and 83.3%, respectively. Conclusion MELD-Lac score has a high prognostic value in HBV-ACLF patients.

Objective To explore the prognostic value of model for end-stage liver disease(MELD)combined with arterial blood lactate(Lac)in admitted patients with hepatitis B virus-associated acute-on-chronic liver failure(HBV-ACLF).Methods Clinical data of 97 cases with hepatitis B virus-associated acute-on-chronic liver failure(HBV-ACLF)admitted to the First Affiliated Hospital of Suzhou University between March 2016 and March 2018 was retrospectively analyzed.Age,gender,complications,MELD score,lactic acid(Lac),total bilirubin(TBil),creatinine(Cr),serum albumin(Alb),high-sensitivity C-reactive protein(CRP),white blood cell count(WBC),platelet count(PLT),hematocrit(Hct),quantification of HBV DNA and HBsAg,and organ support treatment(artificial liver support system,renal replacement therapy and mechanical ventilation)were documented after admission.The primary endpoint of treatment was death due to ineffective medical treatment during hospitalization,abandonment of medical treatment due to deterioration of the health condition,and switch to liver transplantation for patients with poor medical treatment.The risk factors for primary endpoint of treatment were analyzed by binary logistic regression.Hosmer-Lemeshow test was used to evaluate the goodness of fit for the scoring system,and the ROC to predict the prognosis of MELD-Lac.Results Ninety-seven cases with HBV-ACLF were included,56 cases had good prognosis,and 41 cases had bad prognosis(including two cases with poor medical treatment and liver transplantation).The overall improvement rate was 57.7%.MELD score and Lac value in treated group was significantly lower than non-treated group.Bivariable and multivariable logistic regression analysis showed that the MELD score [odds ratio(OR=1.806)],and Lac score [odds ratio(OR=1.820)] was the risk factor for hospitalization and mortality in patients with liver failure(P<0.05).The area under the ROC curve(AUC)and the 95% confidence interval(95% Cl)of prognostic patients with MELD-Lac were significantly better than Lac and MELD scores[0.923(0.84 to 1.00)vs.0.804(0.067 to 0.942)and 0.864(0.75).0.977)],P < 0.05.When the MELD-Lac Youden index was set at 0.746,the optimal threshold was 18.36,and the sensitivity and specificity were 91.3% and 83.3%,respectively.Conclusion MELD-Lac score has a high prognostic value in HBV-ACLF patients.

In this study, the induction of hairy roots of Bupleurum chinense DC. was explored and established after experiments at different conditions: A. rhizogenes A4 was used to infect the leaves bases of B. chinense tube seedlings. The explants were co-cultured on Phytagel-solidified media for 3 days and then, were turned into solid media, similar with the co-culture media except that bacteriostat was added. After 10 days, rootlets began to appear and after 4 to 5 weeks, rootlets can be converted into liquid shaking culture stage. Plants regeneration from hairy root was useful for the research of new germplasm production and the variety improvement breeding. In the present study, the regenerated plants were obtained. One approach was to continuously culture under light conditions the seedlings which parting off spontaneously from the hairy roots during liquid shaking culture. The other approach was to culture the callus-like tissues produced by hairy roots with the optimized regeneration media for the induction of regenerated plants. The results of present study provide a technique to induce hairy roots and plantlet regeneration of B. chinense and this technique is helpful for the researches on metabolism, especially on the Agrobacterium-mediated genetic transformation of B. chinense.

OBJECTIVETo clone the full-length cDNA of a uridine diphosphate glycosyltransferase (UGT) gene which may be involved in saikosaponin biosynthesis of Bupleurum chinense, and construct the transgenic vectors for over expression and RNAi of the cloned UGT. These works will provide foundation for further its function analysis by transgene study.

METHODRAGE and LD-PCR were used to clone the full-length cDNA of the UGT, on the basis of its partial cDNA sequence obtained from our previous 454-sequenced dataset. The ORF and partial sequences of the UGT were PCR cloned using primers with corresponding restriction enzymes cutting sites. The PCR products were digested with corresponding restriction enzymes and then were inserted into pCAMBIA-SUPER 1 300 and pHANNIBAL. The recombinant pHANNIBAL were digested with Not I and then were inserted into a binary vector, pART27. Finally, the transgenic vectors for over expression and RNAi of the cloned UGT were constructed.

RESULTThe full-length cDNA of a UGT were cloned from B. chinense. The recombinant vectors for over expression and RNAi of the UGT were obtained.

CONCLUSIONOur works on full-length cDNA cloning and transgenic vectors construction provide a substantial foundation for follow-up biofunction analysis of the UGT through transgenic research.

Amino Acid Sequence ; Bupleurum ; genetics ; Cloning, Molecular ; Genetic Vectors ; Glucuronosyltransferase ; chemistry ; genetics ; Molecular Sequence Data ; RNA Interference ; Transgenes