As the major member of serine/threonine protein ki-nases family, glycogen synthase kinase 3β ( GSK-3β) has well characterized roles in the development of a variety of diseases. However, it is noticed that modulation of GSK-3β in tumor pro-gress is two-faced. Once the activity of GSK-3βas a“pro-onco-genic factor” is inhibited, opposing role as a“tumor suppressor”can also be disrupted, which will trigger the consequent side effect on activation of Wnt/β-catenin signaling pathway. The is-sue has placed a major obstacle to anti-GSK-3β in cancer treat-ment. In fact, functional compartmentalization of a large number of intracellular signaling events cross-talked with GSK-3β can prevent their mutual interference and determine the cell fate. Therefore, understanding the specific mechanisms of GSK-3β in regulation of diverse signaling systems or refinement of a sub-strate competitive inhibitor may have great significance to exploit approaches selectively target GSK-3β in tumor treatment.

To establish a prokaryotic expression and purification protocol for nuclease P1 (NP1), we first obtained a synthetic NP1 by splicing 22 oligonucleotides with overlapping PCR. We constructed and transformed a secretory expression vector pMAL-p4X-NP1 into Escherichia coli host strains T7 Express and Origami B (DE3) separately. Then, the recombinant NP1 was purified by amylose affinity chromatography, and its activity, thermo-stability and metal-ion dependence were investigated systematically. The results indicated that the expressed fusion proteins MBP-NP1 (Maltose binding protein-NP1) existed mainly in soluble form both in host strains T7 Express and Origami B (DE3), but the specific activity of recombinant protein from Origami B(DE3) strain was higher than T7 Express strain (75.48 U/mg : 51.50 U/mg). When the MBP-tag was cleaved by protease Factor Xa, the specific activity both increased up to 258.1 U/mg and 139.2 U/mg. The thermal inactivation experiments demonstrated that the recombinant NP1 was quite stable, and it retained more than 90% of original activity after incubated for 30 min at 80 degrees C. Zn2+ (2.0 mmol/L) could increase enzyme activity (to 119.1%), on the contrary, the enzyme activity was reduced by 2.0 mmol/L Cu2+ (to 63.12%). This research realized the functional expression of NP1 in the prokaryotic system for the first time, and provided an alternative pathway for NP1 preparation.
Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; genetics ; metabolism ; Fungal Proteins ; biosynthesis ; genetics ; metabolism ; Genes, Synthetic ; Genetic Vectors ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; metabolism ; Single-Strand Specific DNA and RNA Endonucleases ; biosynthesis ; genetics ; metabolism

Objective: To explore determinants of childhood trauma among college students with left-behind experience, and to provide a reference for effective intervention among students with left-behind experience. Methods: A total of 2 468 students selected from 5 universities and 2 higher vocational colleges in tianjin by stratified cluster sampling method were investigated by self-compiled questionnaire and childhood trauma questionnaire. Results: The scores in emotional abuse, sexual abuse, emotional neglect, physical neglect and childhood trauma of students with left-behind experience were significantly higher than those without left-behind experience(t=3.01，3.13，3.24，2.27，3.60，P<0.05)；parental separation times and the frequency of parental return had significant interaction effect on the total score of childhood trauma of students with left-behind experience (F=2.37, P<0.05)；the gender had a significant major effect on the total score of childhood trauma of students with left-behind experience under the interaction with the place of origin, age at first separation,the cumulative time of leftbehind experiences and the frequency of parents contacting (F=4.49，5.23，5.93，5.11，P<0.05)；the age of subjects when parents going out under the interaction with the place of origin, the gender, if only-child,parental separation times and the frequency of parental return；as well as the frequency of parents contacting under the interaction with the place of origin，the household registration, the gender, if only-child and the cumulative time of left-behind experiences also had significantly main effect(F=3.88，4.25，3.32，2.86，3.45；3.82，4.02，2.64，3.29，P<0.05). Conclusion It is necessary to attach great importance to demographic and context information regarding left-behind experiences,which lead to more specific and effective prevention and intervention strategy for individual with left-behind experiences.

Aim To screen the potential inhibitors of post-transcriptional activity of pro-inflammatory media-tor TNF-α from the lipophilic constituents in Chinese Medicine Salvia miltiorrhiza Bunge ( Danshen) , we es-tablished dual luciferase reporter gene system pGL3-TNF-α3′UTR ( 3′untranslated region ) co-transfected with Renilla control gene. Methods Complementary DNA ( cDNA) template was obtained from human um-bilical vein endothelial cells ( HUVECs ) . The full length DNA of TNF-α 3′-UTR was amplified through PCR, and then connected the luciferase reporter vector pGL3-control after enzyme digestion. pGL3-TNF-α 3′UTR constructs were co-transfected with pSVRenilla into the mononuclear macrophages RAW264. 7 cells. The relative activity of reporter genes was measured by dual luciferase reporter ( DLR ) assay system after the stimulus of lipopolysaccharide ( LPS ) in presence or absence of tanshinones compounds. Results The pGL3-TNF-α3′UTR luciferase reporter gene was suc-cessfully constructed. The cloning DNA fragment and sequence were both consistent with the GENBANK da-tabase. LPS significantly induced the relative reporter activityof RAW264 . 7 cells transfected with pGL3-TNF-α 3′UTR. Among four tanshinones compounds, we found only cryptotanshinone could significantly de-crease LPS-induced relative reporter activity. Conclu-sion The pGL3-TNF-α 3′UTR construct combined with DLR assay system was successfully established, which can be used to discover the agents such as cryp-totanshinone that regulate the post-transcription of TNF-α in treatment of inflammatory and malignant dis-eases.

Tumor metastasis is one of the most important biologi-cal characteristics of malignant tumor, and it is also the main factors that cause treatment failure and poor prognosis. Clinical studies have shown that the number of platelets in patients with malignant tumor increased more significantly than that in benign tumor patients and healthy people, which indicate that platelet might be involved in the development process of tumor. Further study found that in the process of cancer spreading to blood, platelet could interact with tumor cells to form tumor emboli, helped tumor cells escape from immune surveillance, thus pro-moted the tumor metastasis. In recent years, related mechanisms on platelets in promoting tumor metastasis were revealed gradual-ly, and several targeted therapies based on platelets were also carried out. This paper reviews the role of platelet in mediating tumor metastasis by hematogenous spread and its mechanisms and discusses the therapy strategies that target platelet, which may provide references for follow-up research and clinical treat-ment.

Objective:To compare the differences in inhaled medication prescriptions among patients with chronic obstructive pulmonary disease (COPD) who visited the Global Initiative for Chronic Obstructive Pulmonary Disease (GOLD 2023) one year after its release and the previous year, and to analyze the impact of GOLD 2023 on physician inhaled medication prescriptions.Methods:This study was a cross-sectional study, with data sourced from the RealDTC study. The study subjects were chronic obstructive pulmonary disease patients who visited the respiratory and critical care departments of 13 hospitals in southern China from November 14, 2021 to November 15, 2023. According to the time of patient visits, they are divided into the following two groups: the group 1 year before the release of GOLD 2023 (November 14, 2021 to November 14, 2022), and the group 1 year after the release of GOLD 2023 (November 15, 2022 to November 15, 2023). We collected demographic characteristics, lung function, symptom scores, history of acute exacerbation in the past year, and inhaled medication prescriptions from patients. According to the symptom score of COPD patients in GOLD 2023 and their history of acute exacerbation in the past year, they were divided into three groups: A, B, and E. The treatment status of inhaled drugs in groups A, B, and E before and after the release of GOLD 2023 was compared.Results:There were statistically significant differences in COPD Assessment Test (CAT) scores, Modified Medical Research Council (mMRC) scores, and the number of acute exacerbations in the past year between patients with COPD before and after the release of GOLD 2023 (all P<0.05). Compared with the group one year before the release of GOLD 2023, the proportion of patients in the group one year after the release of GOLD 2023 using long-acting muscarinic antagonists (LAMA) and inhaled corticosteroids (ICS)+ long-acting β2-receptor agonists (LABA) was lower, while the proportion of patients using LABA+ LAMA and ICS+ LABA+ LAMA was higher (all P<0.05). There was no statistically significant difference ( P>0.05) in the proportion of patients in group A using LAMA between the year before and after the release of GOLD 2023. Compared to the year before the release of GOLD 2023, the proportion of patients in group A who prescribed ICS+ LABA was lower, while the proportion of using LABA+ LAMA and ICS+ LABA+ LAMA was higher (all P<0.05); The proportion of patients in group B who prescribed LAMA and ICS+ LABA was lower (all P<0.05), while the proportion of using LABA+ LAMA and ICS+ LABA+ LAMA was higher (all P<0.05); The proportion of patients in group E who prescribed LAMA and ICS+ LABA was lower (all P<0.05), while the proportion of using LABA+ LAMA and ICS+ LABA+ LAMA was higher (all P<0.05). Conclusions:After the release of GOLD 2023, the prescription of ICS+ LAMA in groups A, B, and E decreased, and the prescriptions of LABA+ LAMA and ICS+ LABA+ LAMA increased compared to before; However, in the real world, the compliance of physicians with GOLD 2023 is still not ideal.

The elaboration of the mechanism and pharmacodynamic substance are the main obstacles to the modernization of Chinese medicine.The rich ingredient of Chinese medicine is almost attention,with a research strategy of forward pharmacology.This strategy often neglects the study of trace components of Chinese medicine.Synthetic lethality,a extremely complex gene interactions,is to magnify the effects of the co-regulation of biological effects (＞ 1 000 times).The theory of synthetic lethality has achieved good results in the development of anti-tumor drugs,including the discovery of PARP inhibitors,the clinical use of chemotherapy drug addition and attenuation combination.In view of this,this research model may be used to elucidate trace effective substance.Based on the reverse thinking of "targetcomponent-effect"and clear synergistic targets,the mechanism of traces and weak-potency substance of traditional Chinese medicine was studied,and the synergistic combination of potential was found.

Objective To review the research of self-management intervention in patients with hematopoietic stem cell transplantation,clarify the intervention elements,so as to provide references for clinical practice and future related research.Methods According to the scoping review framework,a literature search was performed in the PubMed,Embase,CINAHL,Web of Science,Cochrane Library,CNKI,Wan fang Database,VIP,CBMdisc for all the studies on self-management intervention of patients with hematopoietic stem cell transplantation.The retrieval time was from the establishment of the database to December,2022.There were 2 researchers who extracted and analyzed the data of the included literature.Results A total of 12 articles were included,including 3 randomized controlled trials,8 quasi-experimental studies,and 1 mixed study.6 studies were conducted with interventions based on the theoretical basis or nursing model such as self-management oriented 5A nursing model,continuous nursing model,problem-based learning theory.The intervention methods of 12 studies included nurse-led self-management guidance,mobile applications,multidisciplinary collaboration,and graphic cards.The intervention time was mostly 1～3 months.The outcome indicators included feasibility evaluation indicators such as operability,ease of use,and user satisfaction of the application,and effect evaluation indicators such as self-management ability,quality of life,and medication compliance of patients.The results showed that self-management interventions could improve patient self-management ability,quality of life,medication compliance,and improve their psychological and nutritional status.Conclusion The research on self-management intervention in patients with hematopoietic stem cell transplantation is still in the development stage,and its positive effect has been preliminarily verified.Medical staff should strengthen the self-management evaluation of patients with hematopoietic stem cell transplantation,play the role of multidisciplinary team,use electronic health intervention methods,and formulate multi-level,scientific and effective self-management intervention programs.

Traditional Chinese medicine is a treasure of Chinese civilization,which has undergone thousands of years of precipitati-on and clinical verification.According to the theory of traditional Chinese medicine,ancient doctors used natural medicines and natural means to promote self-regulation and damage repair of the body.In recent years,as an important tool to repair body damage,stem cells have set off a research upsurge.With the continuous deepening of stem cell research and the continuous expansion of the fields in-volved,the potential correlation between traditional Chinese medicine and stem cells has attracted more and more domestic and foreign researchers to invest in research.This paper expounds regenerative medicine and stem cells,the advantages of Chinese medicine in regulating stem cells,and the opportunities and challenges faced by Chinese medicine and regenerative medicine,and reviews the cur-rent research trends and frontier trends in this field,in order to provide useful references for subsequent studies.

Recent studies have shown that stress can substantially facilitate breast cancer metastasis,which can be reduced by nonselective β1/β2-adrenergic receptor(β1/β2-AR)blocker.However,several side effects were identified.Thus,it is extremely warranted to explore more effective and better-tolerated β2-AR blocker.Currently,we demonstrated that baicalin(BA),a major bioactive component of Scutellaria bai-calensis Georgi,could significantly attenuate stress hormones especially epinephrine(Epi)-induced breast cancer cell migration and invasion in vitro.Mechanistically,we identified that β2-AR was a direct target of BA via the drug affinity responsive target stability(DARTS)combined with mass spectrum assay,and BA photoaffinity probe with pull-down assay,which was further confirmed by a couple of bio-physical and biochemical assays.Furthermore,we demonstrated that BA could directly bind to the Phe-193 and Phe-289 of β2-AR,subsequently inhibit cyclic adenosine monophosphate-protein kinase A-focal adhesion kinase(cAMP-PKA-FAK)pathway,and thus impede epithelial-mesenchymal transition(EMT),thereby hindering the metastatic progression of the chronic stress coupled with syngeneic and xenograft in vivo orthotopic and tail vein mouse model.These findings firstly identify BA as a potential β2-AR inhibitor in the treatment of stress-induced breast cancer metastasis.