Objective To prepare hepatocyte growth factor(HGF) recombinant protein and confirm its activity preliminarily according to building HGF gene prokaryotic expression vector and transforming into E.coli.Methods Clone HGF inserted into the vector pET-26b(+) to construct prokaryotic expression vector pET-26b(+)-HGF and transform into E.coli Rosseta(DE3).The transformed bacteria induced by IPTG was purified through Ni-NTA resin affinity chromatography frozen-drying after renaturation.Results HGF gene recombinant prokaryotic expression vector pET-26b(+)-HGF was constructed successfully.E.coli Rosseta(DE3) which was transformed into pET-26b(+)-HGF expresses the target protein as the form of inclusion bodies,accounting for 38％ of the total bacterial proteins,and confirmed by Western blot.HGF protein which was purified by Ni-NTA resin affinity chromatography,has a purity of about 95％,and can promote proliferation,migration,and inhibition of apoptosis for human non-small cell lung cancer cell line A549 cells after interaction.Conclusion HGF gene recombinant prokaryotic expression vector pET-26b (+)-HGF was constructed and expressed in transformed E.coli Rosseta(DE3) successfully.They resumed their recombinant HGF protein structure after purification and renaturation,and had biological activity confirmed by in vitro studies.

Objective To understand the potential risk factors influencing the effect of standard anti-tuberculosis (TB) treatment for TB patients co-infected with human immunodeficiency virus (HIV) and provide evidence for the improvement of anti TB therapy.Methods A retrospective study was conducted among 445 TB/HIV patients diagnosed and registered in 7 counties in Yunnan province from January 2010 to June 2012.A structured questionnaire was used to collect the patients' demographic characteristics,diagnosis and treatment information after informed consent.Chi-square test was conducted to compare successful rate of anti TB treatment among the patients with different demographic characteristics.Multivariate logistic regression analysis was conducted to identify risk factors influencing the effect of anti TB treatment.Adjusted OR＞1 means the risk factor of treatment failure.P value less than 0.05 was set as significant level.Results After standard anti TB treatment,397 patients were cured.The five risk factors influencing treatment effect were the existing of 4 suspected TB symptoms when seeking medical care for the first time(adjusted OR=2.208),TB/HIV patients detected in HIV/AIDS screening (adjusted OR=5.856),severe case (adjusted OR=4.607),non-full-course supervision during treatment (in intensive phase adjusted OR=4.129,full-course management adjusted OR=8.090) and interruption of therapy (adjusted OR=21.517).Conclusion Early detection of TB/HIV patients and conducting full course supervision during treatment can improve the effect of anti TB treatment.It is necessary to strengthen the early detection of TB/HIV patients and standarded treatment in Yunnan province.

Objective:To explore the potential molecular mechanism of curcumin in the treatment of post-stroke depression through network pharmacology and molecular docking.Methods:The potential targets of curcumin were searched in GeneCards,PharmGKB and SwissTargetPrediction database.The online Mendelian Inheritance in Man data-base and GeneCards database were searched for potential targets of ischemic stroke and depression.The targets were standardized by Uniprot and then imported into Venn online platform to obtain the intersection targets among the three.Protein-protein interaction(PPI)was visualized by STRING and Cytoscape 3.7.1 software.Metascape database was used to perform Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis of intersection targets.Finally,AutoDock Vina1.5.6 software was used to verify the molecular docking between curcumin and core targets.Results:A total of 1452 potential targets for ischemic stroke and depression,207 drug tar-gets,and 47 intersection targets of the three were obtained.GO functional enrichment analysis yielded 2332 items,and KEGG pathway enrichment analysis yielded 138 items.According to the degree value and enrichment fraction,the core targets such as interleukin-6(IL-6),tumor necrosis factor(TNF),serine/threonine protein kinase 1(AKT1),inter-leukin-1β(IL-1β)and the advanced glycation end products receptor(AGE-RAGE)signaling pathway,interleukin-17(IL-17)signaling pathway,TNF signaling pathway,etc.Molecular docking results showed that curcumin had a strong binding ability to core targets such as IL-6,TNF,AKT1,and 1L-1β.Conclusion:Curcumin may act on key targets such as IL-6,TNF,AKT1,IL-1β,and play an important role through the AGE-RAGE,IL-17,and TNF signaling pathways.

Objective:To systematically evaluate the experience and needs of parental care for premature infants during the transition from hospital to home.Methods:Qualitative studies on the experience and needs of parental care for premature infants transitioning from hospital to home were electronically searched in Cochrane Library, PubMed, Embase, Web of Science, Scopus, PsycINFO, CINAHL, China National Knowledge Infrastructure, VIP, China Biomedical Medline Disc, and WanFang Data. The search period was from the establishment of the database to June 3, 2023. The retrieved studies were evaluated using the quality evaluation criteria for qualitative research of the Joanna Briggs Institute Evidence-Based Health Care Center. The qualitative research results were integrated using the method of aggregation integration.Results:A total of 14 articles were included, 39 research results were extracted, categorized into nine categories. These categories were integrated into three synthesized results, namely the dependency period (parents of premature infants coexisted with joy and concern, and relied on professional support from medical and nursing staff), shock period (parents of premature infants developed negative emotions, faced dual difficulties in home care and role switching, and needed internal and external professional support from the family), adaptation period (parents of premature infants actively responded to changes, sought peer support, and transitioned to the "new normal" of life) .Conclusions:The transition from hospital to home is a dynamic process. During the transition process, parents of premature infants undergo complex emotional experiences and role transitions, requiring support from healthcare professionals, family members, and peers. Medical and nursing workers should pay attention to the psychological changes and diverse needs of parents, provide professional support and guidance to help them adapt to the role of parents, and promote the healthy growth of premature infants.

OBJECTIVETo observe the inhibition of NK4 protein in the proliferation of human Raji lymphoma xenografts in nude mice, and to explore its molecular mechanism.

METHODSModels of human Raji lymphoma xenograft transfected with HGF gene were established by subcutaneous inoculation in nude mice. After establishment of the models, the mice received continuous NK4 protein via tail vein for 4 weeks, and the weight and tumor growth were monitored every week. After 8 weeks, the expression of HGF mRNA and c-Met mRNA of tumor tissues was measured by real-time fluorescent quantitation PCR. The apoptotic index (AI) and microvessel density (MVD) were evaluated by terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) and immunohistochemistry, respectively.

RESULTSThe models of human Raji lymphoma xenograft were successfully established. Although the animal weights of all groups declined, especially in the groups with NK4 protein injection, there was no statistical significance (P > 0.05). The tumor volume in HGF gene transfected group was larger than those of the control groups (P < 0.01), and there was no statistical significance among the control groups (P > 0.05). However, the tumor volume of the NK4 protein injection group decreased significantly (P < 0.01). Expression of HGF mRNA and c-Met mRNA in HGF gene transfected group increased significantly after injection of NK4 protein (P < 0.01). AI in HGF gene transfected group (33.5% ± 12.3%) was significantly lower than that of control groups (89.1% ± 22.3% vs. 81.9% ± 27.0%, P < 0.05), but became significantly higher (119.1% ± 18.9%) after NK4 protein injection (P < 0.01). MVD in HGF gene transfected group (28.5 ± 2.0) was higher than that of control groups (12.2 ± 1.4, 13.8 ± 1.3, P < 0.01), although declined (15.5 ± 2.5) after NK4 protein injection (P < 0.01).

CONCLUSIONSNK4 protein suppresses significantly the growth of human Raji lymphoma xenografts transfected with HGF gene. The pathogenesis may be involved in promoting tumor cell apoptosis and restraining tumor angiogenesis through competitive interrupting HGF/Met signal pathway.

Animals ; Apoptosis ; Hepatocyte Growth Factor ; genetics ; metabolism ; Heterografts ; Humans ; Lymphoma ; genetics ; metabolism ; therapy ; Mice ; Mice, Nude ; Microvessels ; pathology ; Neovascularization, Pathologic ; Proto-Oncogene Proteins c-met ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Signal Transduction ; T-Box Domain Proteins ; administration & dosage ; Transfection ; Transplantation, Heterologous

ObjectiveTo evaluate the pharmacological effect of Buzhong Yiqitang on brain development in offspring of rats with subclinical hypothyroidism （SCH） during pregnancy and explore its potential mechanism. MethodsForty-eight SPF female SD rats were divided into sham operation group （n=8） and model group （n=40）. The rat model of subclinical hypothyroidism （SCH） was constructed by total thyroidectomy combined with postoperative subcutaneous injection of levothyroxine （L-T₄）. The modeled rats were randomly allocated into model， low-， medium-， and high-dose （5.58， 11.16， 22.32 g∙kg^-1， respectively） Buzhong Yiqitang， and euthyrox （4.5×10^-6 g∙kg^-1） groups， with 8 rats in each group. These rats were co-housed with normal male rats for mating. Drug administration started 2 weeks before pregnancy and continued until delivery. Hematoxylin-eosin staining and Golgi-cox staining were used to observe pathological changes in the hippocampal tissue of offspring rats. Western blot was employed to detect the effects of Buzhong Yiqitang on the protein levels of cytochrome C oxidase subunitⅠ （COX）Ⅰ and COXⅣ in the hippocampal tissue of offspring rats. A colorimetric method was used to measure the mitochondrial adenosine triphosphate （ATP） content in the hippocampal tissue of offspring rats. For in vitro experiments， a hydrogen peroxide （H₂O₂）-induced oxidative damage model was established with rat pheochromocytoma cells （PC12）. Interventions included the DNA methyltransferase inhibitor （SGI-1027）， Buzhong Yiqitang-medicated serum， and euthyrox-medicated serum. The cell counting kit-8 （CCK-8） assay was used to examine the effect of Buzhong Yiqitang on cell proliferation. Immunofluorescence staining was performed to evaluate the effect on tubulin beta 3 class Ⅲ （TUBB3） in PC12 cells. Western blot was employed to assess the effects on the protein levels of DNA methyltransferases （TETs and DNMTs） in PC12 cells. The fluorescent probe 2′，7′-dichlorodihydrofluorescein diacetate （DCFH-DA）， luciferase assay， and JC-1 staining were employed to assess the effects of Buzhong Yiqitang on the levels of reactive oxygen species （ROS） and ATP and the mitochondrial membrane potential in PC12 cells. ResultsCompared with the sham group， the model group showed a reduction in the number of hippocampal neurons， incomplete pyramidal cell bodies， loose arrangement， shortened average dendrite length， decreased dendritic complexity and dendritic spine density， and reduced expression levels of COXⅠ and COXⅣ and content of ATP in the brain tissue （P<0.05， P<0.01）. Compared with the model group， after administration of Buzhong Yiqitang and euthyrox， hippocampal neurons exhibited regular arrangement， complete morphology， extended dendrite， increased dendritic complexity and dendritic spine density， and restored expression levels of COXⅠ and COXⅣ and content of ATP （P<0.05， P<0.01）， with the medium-dose Buzhong Yiqitang group showing the best therapeutic effect. In the PC12 cell model of oxidative damage， Buzhong Yiqitang increased the cell viability （P<0.01）， enhanced neuronal differentiation， down-regulated the expression levels of DNMTs （P<0.05）， up-regulated the expression levels of TETs （P<0.05）， decreased the ROS content （P<0.01）， and restored the ATP content and mitochondrial membrane potential （P<0.01）. ConclusionBuzhong Yiqitang protects brain development in offspring of pregnant rats with SCH. It mainly acts on the oxidative stress and mitochondrial dysfunction resulted from abnormal mtDNA methylation， with DNMTs and TETs as the key proteins for its effects.