Objective Shkbp is also called Shkbp1,can competitively inhibit binding CIN85 and c-Cbl,thereby blocking the epidermal growth factor receptor (EGFR) endocytosis and degradation,to play a role in tumor promotion.This study aims to explore the changes in blood cell classification and T cell subsets in blood,bone marrow,and spleen in Shkbp1-deletion (Shkbp-1-/-) mice.Methods Shkbp-1-/-transgenic mice were identified by PCR genotyping.Blood cell classification was performed using an automatic classification system.Flow cytometry was used to detect the T lymphocyte subsets in the blood,bone marrow,and spleen of Shkbp-1-/-and control mice.Results Routine blood examination showed that neutrophils and eosinophils tended to increase and showing significant differences,and there was no significant difference in lymphocytes.The flow cytometry results showed that there was a decrease of CD4+CD8+ double positive cells and increase of bone marrow CD3+ and CD4+ cells in the control group.However,there was a decreasing trend of CD3+,CD4+,CD8+,and CD4+CD8+ cells in the spleen tissues.Conclusions Shkbp1 is involved in the maturation and differentiation of blood cells,and affects the number of immune cells.This study lays a foundation for the study of how Shkbp1 is involved in the differentiation of blood cells.

Objective To investigate the effects of anxiety ,depression and life events on patients with non-erosive gastroesophageal reflux disease (NERD) .Methods From November 2016 to December 2017 ,at The First Affiliated Hospital of the Medical College ,Shihezi University ,135 patients with NERD (NERD group) and 133 volunteers who received physical examination at the same period were enrolled (healthy control group) .The scores of self-rating anxiety scale (SAS) ,self-rating depression scale (SDS ) ,life event scale (LES ) and gastroesophageal reflux disease questionnaire (GerdQ ) of the enrolled individuals were obtained by questionnaire .Mann-Whitney ranking test ,chi square test and Spearman rank correlation test were performed for statistical analysis .Results The patients with anxiety in NERD group accounted for 46 .67％ (63/135) ,which was higher than that in healthy control group (1 .50％ ,2/133) ,and the difference was statistically significant (χ2 =74 .38 , P< 0 .01) .The patients with depression in NERD group accounted for 12 .59％ (17/135) ,which was higher than that of healthy control group (0 ,0/133) ,and the difference was statistically significant (χ2 = 17 .88 ,P< 0 .01) .The SAS score of NERD group was 48 points (45 points ,52 points) ,which was higher than that of healthy control group (37 points (33 points ,43 points)) ,and the difference was statistically significant (Z=-11 .03 ,P<0 .01) .The SDS score of NERD group was 46 points (41 points ,50 points) ,which was higher than that of healthy control group (36 points ,32 points to 40 points) ,and the difference was statistically significant (Z= -11 .03 , P<0 .01) .The LES score of NERD group was 32 points (10 points ,45 points) , which was higher than that of healthy control group (3 points ,0 points to 32 points) ,and the difference was statistically significant (Z= -2 .18 ,P=0 .03) .The score of LES negative events in NERD group was 21 points (3 points ,36 points) ,which was higher than that of the healthy control group (0 points ,0 points to 23 points) , and the difference was statistically significant (Z= -2 .19 , P=0 .03) .GerdQ score was positively correlated with SAS ,SDS ,LES scores and LES negative event score ,and the differences were all statistically significant (r=0 .65 ,0 .60 ,0 .29 and 0 .29 ,all P< 0 .05) .Conclusions Anxiety ,depression and life events are the influencing factors of NERD .The greater the impact of anxiety ,depression and life event (especially negative live events) ,the more severe the NERD symptoms .

Objective To explore the effect and mechanism of necroptosis related proteins in middle cerebral artery occlusion (MCAO) induced brain ischemia/reperfusion injury in mice.Methods C57BL/6 mice were used to establish the brain ischemia/reperfusion injury model induced by MCAO.MCAO mice were treated with z-VAD.fmk (zVAD,1.1 g/kg),GSK'872 (0.7 g/kg) and combined intervention of zVAD and GSK'872,and neurological defect was evaluated by mNSS while brain infarct volume was measured by TTC staining.Western blot and immunofluorescence assay were used to detect protein expression and location of RIP1,RIP3 and MLKL,respectively.Results Neurological defect and brain infarction were caused by MCAO.Compared with MCAO group,zVAD,GSK'872 and the combined intervention alleviated neurological defect and reduced brain infarct volume significantly (P＜0.05 or P＜0.01).The protein levels of RIP3 and RIP1 MLKL were increased in mice of MCAO group,while GSK'872 and the combined intervention obviously downregulated the aforementioned protein expression [RIP1 (GSK'872:0.64± 0.02 vs MCAO:1.28±0.02,P＜0.01);RIP3 (GSK'872:1.08±0.02 vs MCAO:1.45±0.02,P＜0.01);MLKL (GSK'872:0.54±0.01 vs MCAO:1.00±0.01,P＜0.01)].However,zVAD only slightly reduced protein expression of MLKL (P＜0.05) but didn't change the protein expression of RIP1 and RIP3 (P＞0.05).Conclusion RIP1,RIP3 and MLKL are involved in the execution of necroptosis and contribute to the pathological progress of brain ischemia/reperfusion injury.

ObjectiveTo investigate the effects of Yiqi Wenyang Huoxue Lishui components on the cardiac function and mitochondrial energy metabolism in the rat model of chronic heart failure （CHF） and explore the underlying mechanism. MethodsThe rat model of CHF was prepared by transverse aortic constriction （TAC）. Eight of the 50 SD rats were randomly selected as the sham group， and the remaining 42 underwent TAC surgery. The 24 SD rats successfully modeled were randomized into model， trimetazidine （6.3 mg·kg^-1）， and Yiqi Wenyang Huoxue Lishui components （60 mg·kg^-1 total saponins of Astragali Radix， 10 mg·kg^-1 total phenolic acids of Salviae Miltiorrhizae Radix et Rhizoma， 190 mg·kg^-1 aqueous extract of Lepidii Semen， and 100 mg·kg^-1 cinnamaldehyde） groups. The rats were administrated with corresponding agents by gavage， and those in the sham and model groups were administrated with the same amount of normal saline at a dose of 10 mL·kg^-1 for 8 weeks. Echocardiography was used to examine the cardiac function in rats. Enzyme-linked immunosorbent assay was employed to determine the serum levels of N-terminal pro-B-type natriuretic peptide （NT-ProBNP）， hypersensitive troponin（cTnI）， creatine kinase （CK）， lactate dehydrogenase （LD）， free fatty acids （FFA）， superoxide dismutase （SOD）， and malondialdehyde （MDA）. The colorimetric assay was employed to measure the levels of adenosine triphosphate （ATP）， adenosine diphosphate （ADP）， and adenosine monophosphate （AMP） in the myocardial tissue. The pathological changes in the myocardial tissue were observed by hematoxylin-eosin staining and Masson staining. The Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase activities in the myocardial tissue were determined by the colorimetric assay. The ultrastructural changes of myocardial mitochondria were observed by transmission electron microscopy. Western blot was employed to determine the protein levels of ATP synthase subunit delta （ATP5D）， glucose transporter 4 （GLUT4）， and carnitine palmitoyltransferase-1 （CPT-1）. The mitochondrial complex assay kits were used to determine the activities of mitochondrial complexes Ⅰ， Ⅱ， Ⅲ， and Ⅳ. ResultsCompared with the sham group， the model group showed a loosening arrangement of cardiac fibers， fracture and necrosis of partial cardiac fibers， inflammatory cells in necrotic areas， massive blue fibrotic tissue in the myocardial interstitium， increased collagen fiber area and myocardial fibrosis， destroyed mitochondria， myofibril disarrangement， sparse myofilaments， and fractured and reduced cristae. In addition， the rats in the model group showed declined ejection fraction （EF） and fractional shortening （FS）， risen left ventricular end-diastolic diameter （LVIDd）， left ventricular end-systolic diameter （LVIDs）， left ventricular end-diastolic posterior wall thickness （LVPWd）， left ventricular end-systolic posterior wall thickness （LVPWs）， left ventricular end-diastolic volume （LVVOLd）， and left ventricular end-systolic volume （LVVOLs）， elevated levels of NT-ProBNP， cTnI， CK， MDA， FFA， and LD， lowered level of SOD， down-regulated protein levels of GLUT4 and CPT-1， decreased activities of Na⁺-K⁺-ATPase， Ca²⁺-Mg²⁺-ATPase， and respiratory complexes Ⅰ-Ⅳ， and declined levels of ATP5D， ATP， ADP， and AMP （P<0.05， P<0.01）. Compared with the model group， the Yiqi Wenyang Huoxue Lishui components and trimetazidine groups showed alleviated pathological damage of the mitochondria and mycardial tissue， risen EF and FS， declined LVIDd， LVIDs， LVPWd， LVPWs， LVVOLd， and LVVOLs， lowered levels of NT-ProBNP， cTnI， CK， MDA， FFA， and LD， elevated level of SOD， up-regulated protein levels of GLUT4 and CPT-1， increased activities of Na⁺-K⁺-ATPase， Ca²⁺-Mg²⁺-ATPase， and respiratory complexes Ⅰ-Ⅳ， and elevated levels of ATP5D， ATP， ADP， and AMP （P<0.05， P<0.01）. ConclusionYiqi Wenyang Huoxue Lishui components can improve the cardiac function， reduce myocardial injury， regulate glucose and lipid metabolism， optimize the utilization of substrates， and alleviate the damage of mitochondrial structure and function， thus improving the energy metabolism of the myocardium in the rat model of CHF.

Pentostatin is a nucleoside antibiotics with a strong inhibitory effect on adenosine deaminase, and is widely used in the clinical treatment of malignant tumors. However, the high cost hampers its application. In the past 10 years, the biosynthesis of pentostatin were focused on strain breeding, optimization of medium composition and fermentation process. To date, there are no reviews summarizing the elucidated biosynthetic mechanism of pentostatin. This review starts by introducing the various chemical route for production of pentostatin, followed by summarizing the mechanisms of pentostatin biosynthesis in different microorganisms. Finally, challenges for biosynthesis of pentostatin were discussed, and strategies for regulating and improving the microbial synthesis of pentostatin were proposed.
Anti-Bacterial Agents ; Pentostatin

ObjectiveTo systematically review the clinical experience of four sessions of Masters of Traditional Chinese Medicine and two sessions of National Famous Chinese Medicine Practitioners in treating ulcerative colitis （UC）. Data mining and analysis were conducted to clarify the diagnosis and treatment ideas and characteristics of prescription used by these famous doctors in treating UC. MethodsRelevant literature on the treatment of UC by renowned doctors was retrieved from the establishment of the database until March 31， 2023. The literature was collected from databases such as China National Knowledge Infrastructure （CNKI）， Wanfang Data Knowledge Service Platform， Chinese Science and Technology Periodicals Database， and China Biomedical Literature Database. The data mining techniques including frequency analysis， association rules， and cluster analysis were conducted using the Ancient and Modern Medical Case Cloud Platform V2.3.5. ResultsA total of 157 literatures were included in this study， including 115 clinical case data. The study found that UC can be categorized into 14 types of syndrome patterns for treatment， including large intestine dampness-heat syndrome （75，65.22%）， syndrome of dampness stagnancy due to spleen deficiency （23， 20.00%）， spleen-kidney yang deficiency syndrome （21， 18.26%）. The main affected organs were the spleen （85， 73.91%） and large intestine （75， 65.22%）， and they were closely related to liver （24， 20.87%） and the kidney （21， 18.26%）. The predominant pathogenic factors were dampness （83， 72.17%） ， heat （80， 69.57%） and qi deficiency （65， 56.52%）. The treatment involved 30 kinds of treatment methods， including heat-clearing and dampness-draining method （75， 65.22%）， pleen-tonifying and qi-boosting method （25，21.74%） and spleen-invigorating and dampness-transforming method （23， 20.00%）. The medication involved 187 ingredients， with the most commonly used being heat-clearing herbs （37， 19.79%） and tonifying herbs （27， 14.44%）. The tastes of the herbs were mostly sweet （85， 45.45%） ， bitter （80， 42.78%） ， and pungent （71， 37.97%）. The association rules revealed 16 high-frequency combinations mainly composed of Huanglian （黄连）， Baishao （白芍） and Gancao （甘草） along with Baizhu （白术）， Fuling （茯苓）， Muxiang （木香） and Danggui （当归）. ConclusionFamous doctors are skilled in diagnosing and treating UC based on the differentiation of the zang-fu organs and qi-blood. The key pathological mechanism is “spleen deficiency as the root， and large intestine damp-heat as the manifestation”. The core treatment approach is “heat-clearing， spleen-tonifying， and dampness-draining”， with the inclusion of “regulating qi and blood， and balancing cold and heat”.

Cough variant asthma （CVA） is a chronic respiratory disease with cough as its main symptom. The occurrence of CVA is closely related to non-specific airway inflammation， and its pathogenesis involves environmental， genetic， immune， and other factors. In recent years， the advantages of traditional Chinese medicine （TCM） in the treatment of CVA have attracted the attention of experts and scholars in China and abroad， especially its prominent role in regulating immune balance， relieving cough symptoms in CVA patients， and reducing recurrence. T Helper cells 1 （Th1）， T helper cells 2 （Th2）， T helper cells 17 （Th17）， and regulatory T cells （Treg） are derived from CD4⁺ T cells. Immune imbalance of Th1/Th2 and Th17/Treg is a new hotspot in the pathogenesis of CVA and a potential key target in the treatment of CVA by TCM. Th cell subsets are in dynamic balance under physiological conditions， maintaining respiratory immune homeostasis in which pro-inflammatory cytokines and anti-inflammatory cytokines are balanced. Immature helper T cells （Th0） can be differentiated into Th1， Th2， Th17， Treg， and other cell subsets due to cytokine types in the microenvironment in the stage of CVA maturation. The proliferation of Th2 cells leads to eosinophilic airway inflammation. Excessive differentiation of Th17 cells induces neutrophil airway inflammation. Th1/Th2 and Th17/Treg cells are mutually restricted in number and function， and the immune imbalance of Th1/Th2 and Th17/Treg is easy to aggravate the generation of inflammatory response. Restoring immune balance is particularly important for the airway anti-inflammatory therapy of CVA. In this paper， the imbalance of Th1/Th2 and Th17/Treg and the pathogenesis of CVA were systematically expounded. Meanwhile， the latest research on the regulation of immune imbalance by TCM compound， single TCM， and its effective ingredients in the treatment of CVA was reviewed. It provides ideas and references for revealing the scientific connotation of TCM regulating immune balance therapy of CVA， as well as the development of clinical treatment and basic research of CVA.

ObjectiveBased on knowledge mapping， the studies on prediction models in the field of traditional Chinese medicine （TCM） were visually analyzed， which provided a reference basis for the excavation and evolution of the future research direction by combing the development process and summarizing the research hotspots and dynamic trends. MethodChina National Knowledge Infrastructure and Web of Science Core Collection databases were searched to obtain studies on prediction models in the field of TCM from inception to February 28， 2023. Endnote X20 software was used for document management. Knowledge mapping generated by CiteSpace software and VOSviewer software was used to visually analyze the characteristics of publication， institutional cooperation relationship， author cooperation network， co-citation， and keywords. ResultA total of 264 pieces of Chinese literature and 266 pieces of English literature were included， and the overall number of research publications showed an increasing trend year by year. The cooperation relationship between the issuing institutions showed obvious regional characteristics， with the closest cooperation relationship between the universities of TCM and their affiliated hospitals， as well as secondary units subordinate to scientific research institutions. The number of research teams and team members publishing papers in English was higher， and cooperation between different teams was more frequent. Groundbreaking and/or referential studies were widely cited and referred to. The highly cited literature was mainly published in complementary and alternative medicine journals and pharmaceutical journals. Research hotspots mainly focused on clinical prediction models of TCM， quantitative models of TCM， and specific modeling methods. The application of artificial intelligence technologies such as machine learning and deep learning in the field of TCM will be the most cutting-edge research direction in the future. ConclusionThe field of TCM is paying more and more attention to the studies on prediction models， while the research cooperation mode involving multiple organizations and teams has increasingly become the mainstream. With the continuous development of multi-disciplinary integration， studies on prediction models are closely related to the development and rise of innovative techniques and methods， and any breakthrough in theory or application will induce and guide a new round of research upsurge. Systematic reviews of topic-specific prediction models should be carried out in the future to provide evidence-based evidence.

As the most aggressive breast cancer, triple-negative breast cancer (TNBC) is still incurable and very prone to metastasis. The transform growth factor β (TGF-β)-induced epithelial-mesenchymal transition (EMT) is crucially involved in the growth and metastasis of TNBC. This study reported that a natural compound isotoosendanin (ITSN) reduced TNBC metastasis by inhibiting TGF-β-induced EMT and the formation of invadopodia. ITSN can directly interact with TGF-β receptor type-1 (TGFβR1) and abrogated the kinase activity of TGFβR1, thereby blocking the TGF-β-initiated downstream signaling pathway. Moreover, the ITSN-provided inhibition on metastasis obviously disappeared in TGFβR1-overexpressed TNBC cells in vitro as well as in mice bearing TNBC cells overexpressed TGFβR1. Furthermore, Lys232 and Asp351 residues in the kinase domain of TGFβR1 were found to be crucial for the interaction of ITSN with TGFβR1. Additionally, ITSN also improved the inhibitory efficacy of programmed cell death 1 ligand 1 (PD-L1) antibody for TNBC in vivo via inhibiting the TGF-β-mediated EMT in the tumor microenvironment. Our findings not only highlight the key role of TGFβR1 in TNBC metastasis, but also provide a leading compound targeting TGFβR1 for the treatment of TNBC metastasis. Moreover, this study also points out a potential strategy for TNBC treatment by using the combined application of anti-PD-L1 with a TGFβR1 inhibitor.