[摘 要] 目的：探讨烯醇化酶1（ENO1）对胃癌细胞增殖、迁移及侵袭的影响及其分子机制。方法：通过WB法检测ENO1在人胃癌细胞HGC27、MKN-45、N-87、MGC803、BGC-823及人胃黏膜上皮细胞GES-1中的表达水平，利用CRISPR、过表达等基因编辑工具分别构建ENO1敲低及敲低—恢复表达细胞株，将MKN-45和BGC-823细胞分别分组为Ctrl组、ENO1 KD组、ENO1 KD-OE组。通过克隆形成实验、EdU染色、划痕实验、Transwell实验、流式细胞术检测敲低或敲低—恢复表达ENO1对胃癌细胞增殖、迁移、侵袭及凋亡的影响。构建胃癌细胞裸鼠移植瘤模型，通过小动物活体成像技术及肿瘤组织块测量观察ENO1对肿瘤生长的影响。在MKN-45细胞中通过RNA干扰技术沉默ENO1，利用RNA免疫共沉淀-转录组测序（RIP-Seq）结合生物信息学分析技术鉴定ENO1下游的靶基因，分析ENO1调控胃癌细胞增殖、迁移及侵袭的分子机制。结果：ENO1在胃癌细胞中表达水平均显著升高（P < 0.01或P < 0.001）。在MKN-45和BGC-823细胞中敲低ENO1可显著降低胃癌细胞的增殖、迁移、侵袭能力并促进细胞凋亡（P < 0.001或P < 0.000 1）；回复实验结果显示，恢复ENO1的表达后，胃癌细胞的增殖、迁移、侵袭能力均显著提高并减少细胞凋亡（P < 0.05或P < 0.01或P < 0.001或P < 0.000 1）。裸鼠荷瘤实验结果表明，敲低ENO1的表达可以显著抑制裸鼠移植瘤的生长（P < 0.000 1）。与ENO1蛋白具有相互作用的差异基因主要富集于RNA剪切相关通路，且ENO1蛋白与PKM基因具有相互作用，胃癌组织中ENO1与PKM2基因表达事呈正相关（r = 0.886）。结论：ENO1在胃癌细胞中呈高表达，其通过与PKM的前体mRNA相互作用影响PKM的RNA剪接过程，进而调控PKM2的表达水平并促进胃癌的进展。

Objective:To observe the effect of acupuncture combined with vitamin B12 point injection on pain severity in patients with discogenic low back pain and to analyze its potential mechanisms. Methods:A total of 96 patients with discogenic low back pain were randomly divided into two groups.The control group received acupuncture treatment,while the combined group received vitamin B12 point injection in addition to the identical acupuncture treatment in the control group.Both groups were treated for 2 weeks.The visual analog scale(VAS)and Oswestry disability index(ODI)scores were compared before treatment,after 1 week of treatment,and after 2 weeks of treatment.The levels of such serum inflammatory factors as tumor necrosis factor(TNF)-α and interleukin(IL)-6,serum beta-endorphin(β-EP),and prostaglandin(PG)E2 were compared before and after treatment.Adverse reactions and clinical efficacy were compared between the two groups after treatment. Results:The total effective rate in the combined group was higher than that in the control group(P<0.05).The VAS and ODI scores in the combined group after 1 week and 2 weeks of treatment were lower than those in the control group at the same time point(P<0.05).The levels of TNF-α,IL-6,and PGE2 in the combined group were lower than those in the control group after treatment(P<0.05),while the level of β-EP was higher than that in the control group(P<0.05).There was no statistical difference in the overall incidence of adverse reactions between the two groups(P>0.05). Conclusion:Acupuncture combined with vitamin B12 point injection can alleviate pain and promote functional recovery in patients with discogenic low back pain;reducing the levels of TNF-α,IL-6,and PGE2 and increasing the level of β-EP may be part of the mechanism of the therapy.

Objective To explore the value of energy spectrum CT based material imaging for quantitatively and qualitatively evaluating lumbar intervertebral disc degeneration.Methods Lumbar energy spectrum CT and MRI were collected from 82 patients with back and leg pain,and 74 keV CT,water(chondroitin sulfate[CS])and CS(water)material images were obtained.Intervertebral discs Pfirrmann grading(PG)was performed based on MRI.CT values,water(CS)and CS(water)concentration of nucleus pulposus(NP)and annulus fibrosus(AF)in lumbar intervertebral disc with different PG grades were compared,and the relations with PG grades were analyzed.Visual classification of lumbar intervertebral disc was performed based on water(CS)and CS(water)falsecolor map,while PG distribution of different visual classification were compared,and the relations with PG grades were observed.Results Totally 250 lumbar intervertebral discs were enrolled,including 22 of PG Ⅰ,49 of PG Ⅱ,83 of PG Ⅲ and 96 of PG Ⅳ.CT values,water(CS)and CS(water)concentration of anterior AF,NP and posterior AF of intervertebral disc with different PG grades were significantly different(all P＜0.001),and CT values were negatively correlated with PG grades(rs=-0.504,-0.399,-0.258,all P＜0.001),while water(CS)concentration was positively correlated(rs=0.476,0.753,0.324,all P＜0.001)but CS(water)concentration was negatively correlated with PG grades(rs=-0.486,-0.760,-0.329,all P＜0.001).Significant differences of PG grades were found among different water(CS)and CS(water)falsecolor images visual classification(all P＜0.001),while water(CS)and CS(water)falsecolor images visual classification were all positively correlated with PG grades(all rs≥0.700,all P＜0.001).Conclusion Energy spectrum CT based material imaging could be used for quantitatively and qualitatively evaluating lumbar intervertebral disc degeneration.

Objective To explore the diagnosis and clinical characteristics of atypical severe pneumonia caused by Chlamydia abortus(C.abortus).Methods Clinical data of 4 patients diagnosed with atypical severe pneumonia caused by C.abortus in a hospital from January 2021 to November 2022 were collected.Clinical characteristics,dia-gnosis and treatment,and precautions of the disease were comprehensively analyzed.Results All 4 patients were male,aged 63-73 years old,with acute onset,high fever,cough and expectoration.Three patients had a history of contact with poultry,one patient had a history of contact with abortion goat.The interval between the emerging of clinical symptoms and the onset of acute respiratory failure in 4 patients was 1-6 days,and the oxygenation index(PaO2/FiO2)at admission was less than 200 mmHg,which gradually decreased with the progression of the disease,active support with a ventilator was necessary.Two patients had an increase in white blood cell count,4 had an in-crease in neutrophil percentage,3 had a mild decrease in platelet count.Among 4 patients,2,2,3 and 4 patients showed elevated levels of aspartate aminotransferase,alanine aminotransferase,creatine kinase,and serum creati-nine respectively,2 patients had mild hyponatremia,4 patients showed significant increase in C-reactive protein,procalcitonin,and interleukin-6 levels.Four patients'chest CT findings showed main involvement of single or mul-tiple lung lobes,with exudation and consolidation,and later involvement of multiple lobes of lung.The metageno-mic next-generation sequencing of bronchoalveolar lavage fluid detected the DNA sequence of C.abortus.Based on the clinical manifestations,contact history,chest CT,and metagenomic next-generation sequencing results of 4 pa-tients,the diagnosis was C.abortus.atypical severe pneumonia.After timely adjustment of the treatment of anti-in-fection regimen based on doxycycline,the patients'condition improved and were discharged.Conclusion C.abor-tus may also cause human pneumonia,which can lead to serious clinical outcome after infection.Patient had a histo-ry of animal contact should be alert to such diseases.Metagenomic next-generation sequencing can detect C.abortus.

The MADS-box gene family is a very important transcriptional regulator gene, which plays a role in the whole growth and development process of plants. The APETALA1 (AP1) gene is considered to play an important regulatory role in the transformation of plant flowering, but also to control the characteristic development of floral organs. Lonicera macranthoides is used as medicine with dry buds and early flowers. Therefore, studying the potential mechanism of AP1 gene in regulating flower organ development can provide a basis for improving its medicinal value by molecular means. To explore the potential mechanism of the AP1 gene in the regulation of floral organ development in L. macranthoides, the full-length cDNA of the AP1 was cloned by reverse transcription PCR (RT-PCR) and named LmMADS4. The results show that the CDS of the LmMADS4 gene is 729 bp and encodes 242 amino acids, and the LmMADS4 protein contains no signal peptide and no transmembrane structure, which is an unstable hydrophilic protein. Through homologous sequence alignment and phylogenetic analysis, LmMADS4 and L. japonica MADS27 protein cluster into one class and are closely related. Finally, the expression pattern and protein interaction pattern of LmMADS4 were analyzed by real-time reverse transcription-PCR (qRT-PCR) and yeast two-hybrid technology. The qRT-PCR showed that LmMADS4 gene was differentially expressed in the stems, leaves and flower bud at different developmental stages, including bud type variety Longhua and common variety Baiyun; and LmMADS4 gene was highly expressed in the flower buds, and with the development of flower buds, LmMADS4 gene was continuously up-regulated in the flower bud variety Longhua, however, the expression level of LmMADS4 in the Baiyun terminal flower bud was lower than that in the late flower bud, but the difference was not significant. The yeast two-hybrid results showed that the bait vector pGBKT7-LmMADS4 was not toxic to yeast strains and had no self-activating activity. LmMADS4 protein interacted with LmSVP1, LmSVP3 and LmSOC1s proteins. This study can provide a theoretical basis for exploring the mechanism of long flower bud stage and corolla non-unfolding at the molecular level and variety improvement of L. macranthoides.

MADS-box protein family are important transcriptional regulatory factors in plant growth and development. The AGAMOUS 12 (AGL12) subfamily is believed to play an important regulatory role in the process of plant flowering transition. To explore the potential mechanism of AGL12 subfamily involved in regulating the flower development of Lonicera macranthoides, quantitative real-time polymerase chain reaction (qRT-PCR), prokaryotic expression and yeast two-hybrid techniques were used to analyze the expression pattern, protein expression, and protein-protein interaction pattern of LmAGL12 based on transcriptome data. The results showed that the LmAGL12 gene contains a 603 bp open reading frame (ORF), encoding 200 amino acids, and the encoded protein was stable and hydrophilic without a transmembrane region and signal peptide. Through homologous sequence alignment and phylogenetic analysis, it was confirmed that LmAGL12 protein belongs to the MADS-box protein family and is closely related to the AGL12 protein of Heracleum sosnowskyi and Daucus carota subsp. sativus. The LmAGL12 gene was cloned into prokaryotic expression vector pET-28a and the recombinant constructs were transformed into Escherichia coli BL21 (DE3), which inducing the target protein successfully. The yeast two-hybrid results showed that LmAGL12 protein interacts with LmSVP protein, LmSOC1 protein and LmAP1 protein, respectively. The qRT-PCR results showed that LmAGL12 gene were differentially expressed in different development stages of flower bud, stem, and leaves of 'Longhua' and 'Baiyun' in L. macranthoides. The LmAGL12 gene showed the highest expression level in the middle stage of the 'Longhua' floral bud; For the 'Baiyun' variety, the relative expression level of LmAGL12 gene is the highest in the stem. This study cloned the LmAGL12 gene in L. macranthoides and analyzed it expression for the first time, enriching the research on flower organ development and providing a research basis for further exploring the molecular mechanisms of long bud stage and non-unfolded corolla in L. macranthoides, as well as for variety improvement.

Objective:To systematically sort out the development trajectory of the research field of health industry,discuss the research hotspots and frontier topics in depth,and reveal the topics that should be focused on in the future research of the domestic health industry.Methods:Using CiteSpace Bibliometric Visualization Software,1 488 core journal articles in the field of health industry from 1994 to 2023 in CNKI Database were used as research samples to systematically analyze the development trend,research themes,hot issues and cutting-edge research of the health industry.Results:The number of papers published from 1994 to 2023 showed a steady growth trend on the whole.The author-institution partnership was relatively independent and loose,and there was no obvious network of collaboration.Meanwhile,the research hotspots in this field at the current stage mainly focused on the integrated development of"health industry+",health care industry,smart elderly care,health service industry chain,elderly care service system,and the background of healthy China.Conclusion:With the change of residents'cognition and demand for health,the focus of research has begun to transform from the traditional medical treatment model to the new business model of health industry of"managing health,preventing diseases,and focusing on residents'health".Based on the in-depth analysis of the existing research sample literature of the health industry,in order to effectively promote the high-quality and sustainable development of the health industry,the research field should focus on the business model of the health industry in the era of digital economy in the future,formulation and management of industry standards for the health industry,and empirical research at the micro level in the field of health industry research.

Objective:To systematically sort out the development trajectory of the research field of health industry,discuss the research hotspots and frontier topics in depth,and reveal the topics that should be focused on in the future research of the domestic health industry.Methods:Using CiteSpace Bibliometric Visualization Software,1 488 core journal articles in the field of health industry from 1994 to 2023 in CNKI Database were used as research samples to systematically analyze the development trend,research themes,hot issues and cutting-edge research of the health industry.Results:The number of papers published from 1994 to 2023 showed a steady growth trend on the whole.The author-institution partnership was relatively independent and loose,and there was no obvious network of collaboration.Meanwhile,the research hotspots in this field at the current stage mainly focused on the integrated development of"health industry+",health care industry,smart elderly care,health service industry chain,elderly care service system,and the background of healthy China.Conclusion:With the change of residents'cognition and demand for health,the focus of research has begun to transform from the traditional medical treatment model to the new business model of health industry of"managing health,preventing diseases,and focusing on residents'health".Based on the in-depth analysis of the existing research sample literature of the health industry,in order to effectively promote the high-quality and sustainable development of the health industry,the research field should focus on the business model of the health industry in the era of digital economy in the future,formulation and management of industry standards for the health industry,and empirical research at the micro level in the field of health industry research.

Objective:To systematically sort out the development trajectory of the research field of health industry,discuss the research hotspots and frontier topics in depth,and reveal the topics that should be focused on in the future research of the domestic health industry.Methods:Using CiteSpace Bibliometric Visualization Software,1 488 core journal articles in the field of health industry from 1994 to 2023 in CNKI Database were used as research samples to systematically analyze the development trend,research themes,hot issues and cutting-edge research of the health industry.Results:The number of papers published from 1994 to 2023 showed a steady growth trend on the whole.The author-institution partnership was relatively independent and loose,and there was no obvious network of collaboration.Meanwhile,the research hotspots in this field at the current stage mainly focused on the integrated development of"health industry+",health care industry,smart elderly care,health service industry chain,elderly care service system,and the background of healthy China.Conclusion:With the change of residents'cognition and demand for health,the focus of research has begun to transform from the traditional medical treatment model to the new business model of health industry of"managing health,preventing diseases,and focusing on residents'health".Based on the in-depth analysis of the existing research sample literature of the health industry,in order to effectively promote the high-quality and sustainable development of the health industry,the research field should focus on the business model of the health industry in the era of digital economy in the future,formulation and management of industry standards for the health industry,and empirical research at the micro level in the field of health industry research.

Objective:To systematically sort out the development trajectory of the research field of health industry,discuss the research hotspots and frontier topics in depth,and reveal the topics that should be focused on in the future research of the domestic health industry.Methods:Using CiteSpace Bibliometric Visualization Software,1 488 core journal articles in the field of health industry from 1994 to 2023 in CNKI Database were used as research samples to systematically analyze the development trend,research themes,hot issues and cutting-edge research of the health industry.Results:The number of papers published from 1994 to 2023 showed a steady growth trend on the whole.The author-institution partnership was relatively independent and loose,and there was no obvious network of collaboration.Meanwhile,the research hotspots in this field at the current stage mainly focused on the integrated development of"health industry+",health care industry,smart elderly care,health service industry chain,elderly care service system,and the background of healthy China.Conclusion:With the change of residents'cognition and demand for health,the focus of research has begun to transform from the traditional medical treatment model to the new business model of health industry of"managing health,preventing diseases,and focusing on residents'health".Based on the in-depth analysis of the existing research sample literature of the health industry,in order to effectively promote the high-quality and sustainable development of the health industry,the research field should focus on the business model of the health industry in the era of digital economy in the future,formulation and management of industry standards for the health industry,and empirical research at the micro level in the field of health industry research.