OBJECTIVE To investigate the improvement effects and mechanism of Zhichi suanzaoren decoction （ZSD） on hippocampal oxidative stress injury in hippocampal neurons of mice with perimenopausal insomnia. METHODS The potential targets of active ingredients in ZSD were predicted using TCMSP and TCMIP databases； the targets related to insomnia were searched through GeneCards， OMIM and DisGeNET databases； protein-protein interaction network of intersecting targets of ZSD ingredients and insomnia was constructed； Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were conducted on key targets. Sixty mice were divided into sham operation group， model group， ZSD low-， medium-， and high-dose groups （11， 22， and 33 g/kg）， and eszopiclone group （positive control， 1 mg/kg）. Except for sham operation group， the perimenopausal insomnia model was constructed by ovariectomy （OVX） in the other groups. After successful modeling， mice in each group were gavaged with normal saline or the corresponding drug solution， once a day， for three consecutive weeks. The sleep status of mice was evaluated through the pentobarbital sodium sleep synergy experiment， and the pathological changes of hippocampal neurons and the expressions of related genes and proteins in mice were observed by HE staining， immunohistochemistry staining， immunofluorescence staining， transcriptome sequencing technology and Western blot. RESULTS The results of network pharmacology showed that there were 296 intersection targets between ZSD and perimenopausal insomnia. Protein kinase B1 （Akt1） was a key target for treating insomnia with ZSD. After administration of ZSD， the sleep latency of mice was shortened， the sleep duration was prolonged significantly， and the mean optical density value of neuron-specific nuclear protein in the hippocampal CA1 region was significantly increased （P＜0.01）. Additionally， hippocampal neuron damage in OVX mice was significantly alleviated. The results of transcriptome sequencing showed that ZSD significantly upregulated the transcriptional levels of Nfe2l2 gene in hippocampal tissue of OVX mice （P＜0.05）. After administration of ZSD， protein expressions of nuclear factor E2 related factor 2 （Nrf2） and heme oxygenase-1 （HO-1） in hippocampal tissue of OVX mice， as well as the phosphorylated Akt level， were increased significantly （P＜0.01）. CONCLUSIONS ZSD can ameliorate hippocampal oxidative stress injury of hippocampal neurons in perimenopausal insomnia mice by activating the Akt/Nrf2/HO-1 signaling pathway.

This article systematically described the background of the construction of Professor ZHANG Yonghua's theory of emotion-mind pattern identification， the theoretical basis and the ideas of pattern identification and treatment， and its application. The theory is guided by the concept of the unity of body and spirit， focusing on individual endowment and the characteristics of the seven emotions and five minds of the individual， especially the "vexation， constraint， consideration， frightening and sorrow" emotional symptoms， and adherence to the three core disease mechanism "qi， phlegm， fire"， then prescribed based on the patterns， which is suitable for treatment of emotion-mind diseases and related symptoms. For those with "fire" as the main disease mechanism， identify the manifestation of their emotion and mind： patients with impatience， violent rage mostly belong to excess fire， treated with Zhizi Chi Decoction （栀子豉汤）， Longdan Xiegan Decoction （龙胆泻肝汤）； patients with vexation， easy to anger， mostly belong to deficiency fire， treated with Huanglian Ejiao Decoction （黄连阿胶汤）； patients with vexation， constraint anger， mostly belong to constraint fire， treated with Danzhi Xiaoyao Powder （丹栀逍遥散）， Qinlian Wendan Decoction （芩连温胆汤）； patients with overjoy， usually due to exuberance of heart fire， or phlegm clouding the pericardium， treated with Wendan Decoction （温胆汤）， or Diankuang Mengxing Decoction （癫狂梦醒汤）. For those with "phlegm" as the main disease mechanism， accompanied by excessive consideration and thinking， treated with Wendan Decoction. For those with "qi" as the main disease mechanism， Ganmai Dazao Decoction （甘麦大枣汤） is the main treatment for shorter duration of disease， while Banxia Houpo Decoction （半夏厚朴汤） is the main treatment for longer duration of disease.

Objective:To explore the effect of combining hand-brain perception training with hand-brain motor training based on mirror neuron theory on the recovery of upper limb function after a stroke.Methods:A group of 105 stroke survivors with upper limb dysfunction were randomly divided into a hand-brain perception (HP) group, a hand-brain motor (HM) group, and a combination (C) group, each of 35. In addition to conventional rehabilitation treatment (including exercise therapy, occupational therapy and physical factor therapy), the HP and HM groups were given hand-brain perception training and hand-brain motor training respectively, while group C was provided with both. Before the intervention and after 4 weeks, the upper limb motor functioning of all of the participants was assessed using the simplified version of the Fugl-Meyer upper limb motor function scale (FMA-UE). Sensory functioning was quantified using the tactile Semmes Weinstein monofilament examination (SWME), and the modified Barthel index (MBI) was used to quantify the participants′ ability in the activities of daily living.Results:After the intervention the average FMA-UE, MBI and SWME scores of all three groups had improved significantly, with group C′s average FMA-UE and MBI scores significantly better than the other two groups′ averages. The average SWME score of group C was then significantly better than that of group HM.Conclusions:Hand-brain perception combined with hand-brain motor training based on mirror neuron theory can further promote the recovery of upper limb sensory and motor functioning of stroke survivors., Such therapy is worthy of clinical promotion and application.

Recombinase polymerase amplification technology, as a new type of nucleic acid constant temperature amplification detection technology, has shown enormous application potential and development prospects in recent years. Its unique advantages such as high sensitivity, high specificity, low dependence on instruments, and the ability to integrate multiple detection modes make it widely applicable in multiple fields, especially in grassroots and on-site real-time detection. This article reviews the development history, detection principles, research and application progress of recombinant enzyme polymerase amplification technology, and looks forward to its future development, in order to provide useful references for the research and clinical application of rapid pathogen detection methods based on recombinant enzyme polymerase amplification technology.

Integrons, an important system in bacteria that can capture, express, and excise exogenous genes (particularly antibiotic resistance genes), have received considerable attention in recent years. The integrons integrated onto transposons, conjugative plasmids, phages, or chromosomes can capture gene cassettes through site-specific recombination and facilitate their expression. Through a specific excision mechanism, gene cassettes can also be precisely excised and spread within bacterial populations via horizontal gene transfer and other means. This series of processes not only promotes the reconfiguration of gene cassettes and maintains diversity, but also provides flexibility for bacteria to adapt to constantly changing environments. This article will focus on the latest research advances in the capture, excision, and expression regulation mechanisms of integrons, in order to help researchers have a deeper understanding of the evolutionary trends of integrons and provide valuable insights for the study of bacterial antibiotic resistance dissemination mechanisms and the development of new antibiotics.

Objective:To establish of a newmethod:for the rapid detection of H5N1-Avian Influenza virus by combining reverse transcription (RT), multiple cross displacement amplification (MCDA) and nanoparticle-based lateral flow biosensor (LFB).Methods:MCDA primers were designed based on gene sequences specific to Hemagglutinin (HA) and Neuraminidase (NA) in H5N1 avian influenza virus. The target genes HA and NA were amplified through reverse transcription and MCDA in one reaction system. Results were displayed by LFB. The assay was named as H5N1-mRT-MCDA-LFB. The reaction conditions of the H5N1-RT-MCDA-LFB method were optimized, and the sensitivity and specificity were also assessed.Results:The H5N1-RT-MCDA-LFB assay could achieve good amplification effect at a constant temperature of 65 ℃ for 40 minutes. The method had a lower limit of detection of 100 fg per reaction with 100-fold higher sensitivity than that of the RT-qPCR (lower limit of detection 10 pg per reaction). The assay was negative in detecting 28 common viruses, mycoplasmas, chlamydias, bacterias and funguses, except for H5N1. In addition, the H5N1-RT-MCDA-LFB method showed better validation in simulated clinical samples with a lower limit of detection at 1×10 2 copies/ml. Conclusion:The H5N1-RT-MCDA-LFB assay is a valuable molecular diagnostic technique for detecting H5N1 avian influenza virus due to its simplicity, rapidity, sensitivity and specificity.

[Objective]To summarize Professor ZHANG Youhua's clinical experience in treating multiple-dream disorder based on the emotion syndrome differentiation theory.[Methods]By sorting out,reviewing and analyzing Professor ZHANG's medical cases related to the treatment of multiple dream disorder,this paper summarized Professor ZHANG's academic experience in the treatment of multiple-dream disorder from the aspects of etiology,pathogenesis,treatment principles and methods of traditional Chinese medicine understanding the association between dreams and emotional states,and listed one medical case for vevification.[Results]In the treatment of multiple dream disorder,Professor ZHANG emphasizes the importance of the emotional experience in dreams rather than the content of the dreams themselves.Therefore,the use of emotion syndrome differentiation method is deemed appropriate.According to the emotional experience in dreams,multiple-dream disorder is divided into five basic types:thinking,panic,anger,sad sorrow and joy syndromes.The three core pathogeneses of"Qi,phlegm and fire"are addressed through methods such as eliminating phlegm,regulating Qi and clearing fire.The medical case cited was multiple-dream disorder with panic syndrome,"phlegm stagnation and fire hyperactivity"was the main pathogenesis,the treatment of clearing fire and eliminating phlegm,calming the heart and tranquilizing the mind was adopted,and the prescription was based on Qinlian Wendan Decoction,added and subtracted according to the syndrome,and achieved a significant effect.[Conclusion]Professor ZHANG Yonghua's approach to treating multiple-dream disorder is based on the theory of emotional syndrome differentiation theory.He emphasizes the importance of emotional experiences in dreams,and his unique experience has shown remarkable clinical results.This approach is worth considering,learning from and promoting.

Objective To explore the medication experience of famous traditional Chinese medicine(TCM)Zhang Yonghua in insomnia based on the data mining and analysis,and to provide guidance for the treatment of insomnia with traditional Chinese medicine.Methods A total of 195 medical cases of famous TCM for insomnia disorder were collected.The Ancient and Modern Medical record cloud platform V 2.3.7 was used to analyze the TCM syndromes,the frequency of Chinese medicinals,formulas,four properties,five flavors and the meridian tropism of Chinese medicinals,and hierarchical cluster analysis,community analysis and complex network analysis were also carried out to summarize the medication rules of TCM Zhang Yonghua for insomnia.Results The main TCM syndrome types of insomnia were irritability and irritability and excessive thinking.A total of 115 Chinese medicinals were involved in 195 cases,with the main actions of calming the mind,clearing fire,resolving phlegm and regulating thi.Complex network analysis obtained the core traditional Chinese medicines as bupleurum,Scutellaria baicalensis,Pinellia pinellia,Poria cocos,Shouwu vine,Acacia bark,Coptis chinensis,Yuanzhi,Jujube kernel and Licorice.Conclusion Famous TCM Zhang Yonghua treated insomnia with the emotional syndrome differentiation theory.According to the pathogenesis of,phlegm,and fire,formulas of regulation,phlegm resolving,and fire clearing are prescribed respectively for its root cause,formulas of calming the mind a given for the manifestation,and finally to cure insomnia by regulating abnormal emotions.

Objective To observe the effect of traditional Chinese medicine(TCM)syndrome differentiation treatment of insomnia on depressive symptoms and interleukin(IL)-6 level and tumor necrosis factor(TNF)-α level in serum.And then to discuss the effect of TCM syndrome differentiation treatment of insomnia on reducing the occurrence and development of depression.Methods Sixty patients with insomnia disorder in Hangzhou Hospital of Traditional Chinese Medicine from September 2020 to July 2022 were selected as research objects,and they were divided into intervention group and blank group 30 patients for each group,according to their treatment intention and they were treated with sleep hygiene education plus TCM syndrome differentiation treatment and sleep hygiene education alone respectively.The changes of sleep,depression,serum IL-6 level and TNF-α level were compared between two groups.Results After 2 weeks of intervention,the intervention group's Pittsburgh sleep quality index score,patient health questiomnare-9 score,TCM symptom score,and serum IL-6 and TNF-α level were all significantly reduced,and the difference was statistically significant(P＜0.05).Conclusion TCM syndrome differentiation treatment can effectively improve insomnia and depression,thus delaying the occurrence and development of depression,and is an effective method for early intervention of depression.

Objective To investigate the expression levels of FOXO1 and SMAD4 in esophageal cancer (EC) tissues and their associations with clinicopathological features and prognosis. Methods Tissue samples of cancerous and adjacent non-cancerous tissues were collected from 131 EC patients. Hematoxylin-eosin (HE) staining was performed to observe the pathological morphology of EC and adjacent tissues. The mRNA expression of FOXO1 and SMAD4 was measured using quantitative real-time polymerase chain reaction (qRT-PCR), while the protein expression of FOXO1 and SMAD4 was determined by immunohistochemistry; Spearman's correlation analysis was employed to assess the correlation between FOXO1 and SMAD4 protein expression in EC tissues; Cox regression analysis was conducted to analyze factors influencing the prognosis of EC patients; Kaplan-Meier analysis was used to perform survival analysis for EC patients. Results The expression of FOXO1 mRNA and its protein positive expression rate were significantly higher in the EC tissues than those in the adjacent tissues, whereas the expression of SMAD4 mRNA and its protein positive expression rate were significantly lower (P < 0.05). FOXO1 and SMAD4 were associated with tumor diameter, tumor differentiation, lymph node metastasis and TNM stage (P < 0.05). A negative correlation was observed between FOXO1 and SMAD4 protein expression in EC tissues (r=-0.419, P < 0.05). FOXO1, SMAD4, tumor diameter, differentiation, lymph node metastasis and TNM stage were identified as factors influencing mortality in EC patients (P < 0.05). The 3-year overall survival rate of EC patients was 84.73% (111/131). The 3-year cumulative survival rate was significantly lower in the EC patients with positive FOXO1 expression than in thosewith negative FOXO1 expression (P < 0.05), while it was significantly higher in patients with positive SMAD4 expression than in those with negative SMAD4 expression (P < 0.05). Conclusion The positive expression rate of FOXO1 is higher, while that of SMAD4 is lower in EC tissues. Their protein expressions were associated with clinicopathological features and prognosis in EC patients. FOXO1 and SMAD4 may serve as important biomarkers for predicting the prognosis of EC.