Guided by the turbidity toxin theory， it is believed that the key pathogenesis of constipation-predominant irritable bowel syndrome is the obstruction of turbidity toxin and the disruption of intestinal function. Treatment is based on the principles of dispelling turbidity toxin and promoting intestinal function. The clinical patterns can be divided into three types， turbidity toxin heat accumulation pattern， turbidity toxin combined with liver depression and qi stagnation pattern， and turbidity toxin combined with qi and yin deficiency pattern. The treatment can respectively use self-prescribed Tongfu Jiangzhuo Formula （通腑降浊方） to clear heat and unblock the bowels， direct the turbid downward and resolve toxins； use self-prescribed Shugan Jiangzhuo Formula （疏肝降浊方） to soothe the liver and move qi， direct the turbid downward and resolve toxins； use self-prescribed Mazhi Jiangzhuo Formula （麻枳降浊方） to boost qi and nourish yin， moisten the intestines to remove turbidity and resolve toxins.

ObjectiveTo develop the Evidence Grading Scale for Ancient classical prescriptions in Traditional Chinese medicine， assess its reliability and validity， and apply it in practice to provide multi-source evidence for clinical practice guidelines development. MethodsLiterature retrieval was conducted to extract and screen existing evaluation dimensions， then the initial items were summarized using thematic analysis. Experts in the clinical medicine， medical history and literature participated in the Delphi questionnaire survey to evaluate and refine the items. An expert consensus meeting was conducted to finalize the included items， refine the method for items evaluation and evidence grading. The evidence quality rating scale for ancient classical traditional Chinese medicine （TCM） prescriptions was then established and tested for reliability and validity. ResultsThrough literature review， extraction， screening and summarization， a total of 3 dimensions and 12 initial items were formed. Questionnaires were sent to 69 experts to evaluate the initial items， with a questionnaire response rate of 100% and an expert authority coefficient of 0.92. All 12 items were retained for they had importance scores above 4. The Evidence Grading Scale on Ancient classical prescriptions in Traditional Chinese medicine includes 3 dimensions with 12 items. The 3 dimensions includes ancient evidence， inheritance status， and modern application. Each dimension contains 4 items， and each item has a full score of 5 points. The evidence was rated as high-level， moderate-level， and low-level according to the final scores. The content validity index （CVI） of the 12 items was ＞0.9， the average CVI of the scale was 0.98， and the intraclass correlation coefficient （ICC） was 0.90. ConclusionThe Evidence Grading Scale on Ancient classical prescriptions in Traditional Chinese medicine has good reliability and validity， which is practical for use in the development of TCM clinical guidelines and can better support clinical decision-making.

The clinical manifestations of colorectal polyps are consistent with the characteristics of dampness, stickiness and heaviness. The TCM constitutions in the prone population are mostly related to dampness. The pathological changes of intestinal flora imbalance, intestinal micro inflammation, neuroendocrine immune network and abnormal aquaporin in colorectal polyps are consistent with the research results of modern mechanism of dampness pathogen. This article believed that the TCM pathogenesis of colorectal polyps caused by damp pathogen is the accumulation of spleen deficiency and dampness caused by improper diet, poor emotion and other factors, and the interweaving of various diseases and pathogens to form tangible foreign bodies. According to the pathogenic characteristics of damp pathogen and the pathogenic factors of colorectal polyps, the influence of damp pathogen on the pathogenesis of colorectal polyps was discussed, in order to provide an effective TCM theoretical basis for the diagnosis and treatment of colorectal polyps in clinic.

Based on the theory of "mutual interference of clear and turbid Qi" in Huangdi Neijing（《黄帝内经》）， this study explored the pathogenesis of spleen and stomach diseases and the therapeutic effects of Banxia Xiexintang on them. It suggested that "mutual interference of clear and turbid Qi" represents a pathological state of Yin and Yang disturbance and imbalance in Qi circulation due to the mixture of clear and turbid Qi， which can elucidate the pathogenesis of spleen and stomach diseases. According to this theory， the pathogenesis of spleen and stomach diseases was summarized as Qi disorder in spleen and stomach， disharmony between Ying Qi and Wei Qi， and conflict between cold and heat. Banxia Xiexintang， as a crucial prescription for treating spleen and stomach diseases， achieves its therapeutic effects by dispersing stagnation with pungent flavor， descending adverse Qi with bitterness， regulating Ying Qi and Wei Qi， and harmonizing cold and heat. By regulating Qi circulation， balancing internal and external factors， and addressing deficiency and excess， it can rectify the pathological state of "mutual interference of clear and turbid Qi" of spleen and stomach diseases. Modern research reveals that Banxia Xiexintang can modulate gastrointestinal motility， restore mucosal immune barrier function of the digestive system， and exhibit optimal therapeutic effects when combined with both cold-cool and warm-hot medicines， aligning with its therapeutic role under the theory of "mutual interference of clear and turbid Qi". By delving into the essence of the "mutual interference of clear and turbid Qi" theory and exploring the pathogenesis of spleen and stomach diseases and the therapeutic effects of Banxia Xiexintang based on this theory， this study further elucidated the inherent connection between spleen and stomach diseases and the "mutual interference of clear and turbid Qi" theory， offering insights and theoretical references for the clinical diagnosis and treatment of spleen and stomach diseases.

Background::Pulmonary embolism (PE) is a severe and acute cardiovascular syndrome with high mortality among patients with autoimmune inflammatory rheumatic diseases (AIIRDs). Accurate prediction and timely intervention play a pivotal role in enhancing survival rates. However, there is a notable scarcity of practical early prediction and risk assessment systems of PE in patients with AIIRD.Methods::In the training cohort, 60 AIIRD with PE cases and 180 age-, gender-, and disease-matched AIIRD non-PE cases were identified from 7254 AIIRD cases in Tongji Hospital from 2014 to 2022. Univariable logistic regression (LR) and least absolute shrinkage and selection operator (LASSO) were used to select the clinical features for further training with machine learning (ML) methods, including random forest (RF), support vector machines (SVM), neural network (NN), logistic regression (LR), gradient boosted decision tree (GBDT), classification and regression trees (CART), and C5.0 models. The performances of these models were subsequently validated using a multicenter validation cohort.Results::In the training cohort, 24 and 13 clinical features were selected by univariable LR and LASSO strategies, respectively. The five ML models (RF, SVM, NN, LR, and GBDT) showed promising performances, with an area under the receiver operating characteristic (ROC) curve (AUC) of 0.962-1.000 in the training cohort and 0.969-0.999 in the validation cohort. CART and C5.0 models achieved AUCs of 0.850 and 0.932, respectively, in the training cohort. Using D-dimer as a pre-screening index, the refined C5.0 model achieved an AUC exceeding 0.948 in the training cohort and an AUC above 0.925 in the validation cohort. These results markedly outperformed the use of D-dimer levels alone.Conclusion::ML-based models are proven to be precise for predicting the onset of PE in patients with AIIRD exhibiting clinical suspicion of PE.Trial Registration::Chictr.org.cn: ChiCTR2200059599.

Marsdenia tenacissima injection, a standard Marsdenia tenacissima extract (MTE), has been approved as an adjuvant therapeutic agent for various cancers. Our previous study showed that MTE inhibited the proliferation and metastasis of prostate cancer (PCa) cells. However, the underlying mechanisms and active ingredients of MTE against PCa were not completely understood. This study revealed that MTE induced significant decreases in cell viability and clonal growth in PCa cells. In addition, MTE induced the apoptosis of DU145 cells by reducing the mitochondrial membrane potential and increasing the expression of Cleaved Caspase 3/7, Cyt c, and Bax. In vivo, DU145 xenografted NOD-SCID mice treated with MTE showed significantly decreased tumor size. TUNEL staining and Western blot confirmed the pro-apoptotic effects of MTE. Network pharmacology analysis collected 196 ingredients of MTE linked to 655 potential targets, and 709 PCa-associated targets were retrieved, from which 149 overlapped targets were screened out. Pathway enrichment analysis showed that the HIF-1, PI3K-AKT, and ErbB signaling pathways were closely related to tumor apoptosis. Western blot results confirmed that MTE increased the expression of p-AKT^Ser473 and p-GSK3β^Ser9, and decreased the expression of p-STAT3^Tyr705in vitro and in vivo. A total of 13 compounds in MTE were identified by HPLC-CAD-QTOF-MS/MS and UPLC-QTOF-MS/MS. Molecular docking analysis indicated that six compounds may interact with AKT, GSK3β, and STAT3. In conclusion, MTE induces the endogenous mitochondrial apoptosis of PCa by regulating the AKT/GSK3β/STAT3 signaling axis, resulting in inhibition of PCa growth in vitro and in vivo.
Mice ; Animals ; Male ; Humans ; Mice, Inbred NOD ; Mice, SCID ; Marsdenia ; Proto-Oncogene Proteins c-akt ; Glycogen Synthase Kinase 3 beta ; Molecular Docking Simulation ; Phosphatidylinositol 3-Kinases ; Tandem Mass Spectrometry ; Prostatic Neoplasms ; Apoptosis ; STAT3 Transcription Factor

ObjectiveTo explore the effect of Qingre Huashi Tiaoshu prescription on mice with ulcerative colitis （UC） based on the change of the intestinal mucosal barrier function. MethodSix of thirty-six C57BL/6 mice were randomly selected as the normal group. The remaining mice were free to drink 2.5% dextran sulfate sodium （DSS） solution for 7 days to prepare UC model mice. After modeling， they were randomly divided into model group， mesalazine group， and Qingre Huashi Tiaoshu prescription high-， medium- and low-dose groups， 6 in each group. Mesalazine group was given 0.52 g·kg^-1·d^-1 suspension （ig）. Qingre Huashi Tiaoshu prescription high-， medium- and low-dose groups were given 3.38， 1.69， 0.845 g·kg^-1·d^-1 suspension， respectively （ig）. The normal group and the model group were treated with an equal volume of purified water for 10 days （ig）. After intervention， the general condition， body weight， disease activity index （DAI） score， and colon length of the 6 groups of mice were compared. The colon tissue was stained with hematoxylin and eosin （HE） for pathological analysis， and the expression of Ki67 in colon tissue was detected by immunohistochemistry. Western blot was used to determine the protein expression of matrix metalloproteinase-9 （MMP-9）， cleaved cysteine aspartate-specific protease-3 （Caspase-3） and Caspase-3 in colon tissue. ResultCompared with the model group， the mesalazine group and the Qingre Huashi Tiaoshu prescription high- and medium-dose groups alleviated the diarrhea， blood in the stool， weight loss， and other symptoms in UC mice， reduced the DAI score （P<0.05）， and restored the colon shape （P<0.05）. The colonic mucosal defect and inflammatory cell infiltration were improved in the mesalazine group and the Qingre Huashi Tiaoshu prescription high- and medium-dose groups，and there were more intact crypt stem cells. Western blot showed that the protein expression of MMP-9 and cleaved Caspase-3 in the mesalazine group and the Qingre Huashi Tiaoshu prescription high-dose group was lower than that in the model group （P<0.05）， and there was no significant difference between the groups. The expression of Caspase-3 in the Qingre Huashi Tiaoshu high-dose prescription group was decreased compared with the condition in the model group （P<0.05）. ConclusionQingre Huashi Tiaoshu prescription could effectively relieve DSS-induced UC， and its mechanism of action might be related to regulating the expression of MMP-9， cleaved Caspase-3， and Caspase-3， and promoting the repair of intestinal mucosal barrier.

In order to improve the management level of medicine list of medical institutions in China ，and help medical institutions build a medicine list of medical institutions with reasonable drug use structure ，standardized adjustment procedures ， convenient operation and application and scientific evaluation methods ，so as to meet the needs of clinical rational drug use to the greatest extent ，with the support of the Pharmaceutical Care Professional Committee of the Chinese Pharmaceutical Association ， China-Japan Friendship Hospital and the First Affiliated Hospital of University of Science and Technology of China （Anhui Provincial Hospital ）jointly initiate and complete Guideline for the Evaluation of Medicine List in Chinese Medical Institutions jointly with a number of medical institutions. In strict accordance with the methodological requirements of World Health Organization standard guidelines ，based on the Delphi method ，the guideline formulation working group has constructed the quality evaluation index system and quantitative scoring table of medicine list management in medical institutions from the 5 dimensions of organization and management ，structure，adjustment，application and e valuation of the list. It is used to help medical institutions evaluate the quality of their medicine list management ，so asto play a positive role in the fine management of medicine list in medical institutions.

Enzymes and cell factories are the core of industrial biotechnology. They play important roles in various fields such as medicine, chemical industry, food, agriculture, and energy. Usually, natural enzymes and cells need to be engineered to improve the catalytic efficiency, stability and enantioselectivity. Directed evolution makes it possible to rapidly improve the properties of enzymes and cell factories. Sensitive and reliable high-throughput screening approaches are the key for successful and efficient engineering of enzymes and cell factories. In this review, we first summarize the advantages and disadvantages of different screening methods and signal generation strategies as well as their application scope; we then describe the latest advances of ultra-high throughput screening technology applied in the directed evolution of enzymes and cell factories in the past three years. On this basis, we discuss the limiting factors that need to be further improved for high-throughput screening systems and forecast the future development trends of high-throughput screening methods, hoping that researchers in various fields including biotechnology and instrument development can cooperate closely to enhance the reliability and applicability of the high-throughput screening techniques.
Biotechnology ; Directed Molecular Evolution ; Enzymes ; High-Throughput Screening Assays ; Reproducibility of Results

Objective:To investigate the variation trend of peripheral blood CD34 + cells during the hematopoietic stem cell mobilization and its influence on the collection timing and results. Methods:The clinical data of 62 patients with hematologic diseases undergoing autologous peripheral blood hematopoietic stem cell mobilization from April 2012 to March 2017 in Shanxi Provincial Cancer Hospital were analyzed. Mobilization regimen used chemotherapy combined with granulocyte colony-stimulating factor (G-CSF) to monitor the number of white blood cells (WBC), mononuclear cells (MNC), CD34 + cells in peripheral blood and apheresis concentrates, and the correlation with CD34 + cells was analyzed. Furthermore, the receiver operating characteristic (ROC) curve was used to establish the threshold to start apheresis. Results:MNC (5.66±1.11)×10 8/kg and CD34 + cell count (2.15±1.20)×10 6/kg were obtained in 62 patients who received 136 times collection in total. The peak of peripheral blood CD34 + cells count appeared at day 4-5 after the treatment of G-CSF, and then it went down. CD34 + cell count in the product was correlated with the peripheral blood CD34 + cell count collected on the day ( r = 0.879, P < 0.01), and it was also correlated with the peripheral blood WBC and MNC collected on the day as well as MNC count in the product (all P < 0.05). Furthermore, the ROC curve analysis demonstrated that peripheral blood CD34 + cells count > 23/μl was the optimal threshold for stem cell collection on the day, 85.2% of patients reaching up to the threshold could be successfully collected at one time. Conclusions:The variation trend of peripheral blood CD34 + cell count can guide the best time of stem cell collection in clinic. Peripheral blood CD34 + cell count is the reliable index to predict CD34 + cells count in the products. Peripheral blood CD34 + cells count > 23/μl could be used as the collection threshold.