Objective:To analyze the differential genes and related signaling pathways of microglia subpopulations in Parkinson's disease (PD) -like mouse brains induced by paraquat (PQ) based on single-cell RNA sequencing, and provide clues to elucidate the mechanism of PQ-induced PD-like changes in the brain of animals.Methods:In September 2021, six male 6-week-old C57BL/6 mice were randomly divided into control group and experimental group (three mice in each group) . The mice were injected with saline, 10.0 mg/kg PQ intraperitoneally, once every three days, and 10 consecutive injections were used for modeling. After infection, the brains of mice were taken and 10×Genomics single-cell RNA sequencing was performed. Microglia subpopulations were screened based on gene expression characteristics, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed. The differential genes of microglia subpopulations between the experimental group and control group were further screened, and functional enrichment analysis was performed using bioinformatics tools. Mouse microglia (BV2 cells) were treated with 0, 60, 90 μmol/L PQ solution, respectively. And real-time fluorescence quantitative PCR experiments were conducted to validate the expressions of differential genes hexokinase 2 (Hk2) , ATPase H+ Transporting V0 Subunit B (Atp6v0b) and Neuregulin 1 (Nrg1) .Results:Cluster 7 and Cluster 20 were identified as microglia subpopulations based on the signature genes inositol polyphosphate-5-phosphatase d, Inpp5d (Inpp5d) and transforming growth factor beta receptor 1 (Tgfbr1) , and they reflected the microglia-activated M2 phenotype. The bioinformatics analysis showed that the characteristic genes of identified microglia subpopulations were enriched in endocytosis. In terms of molecular function, it mainly enriched in transmembrane receptor protein kinase activity and cytokine binding. The up-regulated genes of Cluster 7 were mainly enriched in lysosomal pathway, endocytosis pathway, and down-regulated genes were mainly enriched in neurodegenerative disease and other signaling pathways. The up-regulated genes of Cluster 20 were mainly enriched in signaling pathways related to PD, and down-regulated genes were mainly enriched in cyclic adenosine 3', 5'-monophosphate (cAMP) signaling pathways, neurological development, synaptic function and other signaling pathways. The results of real-time fluorescence quantitative PCR showed that the expressions of Hk2 mRNA and Atp6v0b mRNA increased and the expression of Nrg1 mRNA decreased in the 90 μmol/L PQ-treated BV2 cells compared with the 0 μmol/L, and the differences were statistically significant ( P<0.05) . Conclusion:Microglia are activated in the PQ-induced PD-like mouse model and polarized toward the M2 phenotype. And their functions are associated with lysosomal (endocytosis) , synaptic functions and the regulation of PD-related pathways.

Objective:To analyze the differential genes and related signaling pathways of microglia subpopulations in Parkinson's disease (PD) -like mouse brains induced by paraquat (PQ) based on single-cell RNA sequencing, and provide clues to elucidate the mechanism of PQ-induced PD-like changes in the brain of animals.Methods:In September 2021, six male 6-week-old C57BL/6 mice were randomly divided into control group and experimental group (three mice in each group) . The mice were injected with saline, 10.0 mg/kg PQ intraperitoneally, once every three days, and 10 consecutive injections were used for modeling. After infection, the brains of mice were taken and 10×Genomics single-cell RNA sequencing was performed. Microglia subpopulations were screened based on gene expression characteristics, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed. The differential genes of microglia subpopulations between the experimental group and control group were further screened, and functional enrichment analysis was performed using bioinformatics tools. Mouse microglia (BV2 cells) were treated with 0, 60, 90 μmol/L PQ solution, respectively. And real-time fluorescence quantitative PCR experiments were conducted to validate the expressions of differential genes hexokinase 2 (Hk2) , ATPase H+ Transporting V0 Subunit B (Atp6v0b) and Neuregulin 1 (Nrg1) .Results:Cluster 7 and Cluster 20 were identified as microglia subpopulations based on the signature genes inositol polyphosphate-5-phosphatase d, Inpp5d (Inpp5d) and transforming growth factor beta receptor 1 (Tgfbr1) , and they reflected the microglia-activated M2 phenotype. The bioinformatics analysis showed that the characteristic genes of identified microglia subpopulations were enriched in endocytosis. In terms of molecular function, it mainly enriched in transmembrane receptor protein kinase activity and cytokine binding. The up-regulated genes of Cluster 7 were mainly enriched in lysosomal pathway, endocytosis pathway, and down-regulated genes were mainly enriched in neurodegenerative disease and other signaling pathways. The up-regulated genes of Cluster 20 were mainly enriched in signaling pathways related to PD, and down-regulated genes were mainly enriched in cyclic adenosine 3', 5'-monophosphate (cAMP) signaling pathways, neurological development, synaptic function and other signaling pathways. The results of real-time fluorescence quantitative PCR showed that the expressions of Hk2 mRNA and Atp6v0b mRNA increased and the expression of Nrg1 mRNA decreased in the 90 μmol/L PQ-treated BV2 cells compared with the 0 μmol/L, and the differences were statistically significant ( P<0.05) . Conclusion:Microglia are activated in the PQ-induced PD-like mouse model and polarized toward the M2 phenotype. And their functions are associated with lysosomal (endocytosis) , synaptic functions and the regulation of PD-related pathways.

Endoscopic resection has become one of the standard treatment options for cT1N0M0 esophageal cancer owing to the wide-spread use of digestive endoscopy.However,patients who undergo non-curative resection,as confirmed by postoperative pathology invest-igations,encounter complex decisions regarding follow-up management.This paper reviews the advancements in domestic and internation-al research on non-curative endoscopic resection of cT1N0M0 esophageal cancer,highlighting the controversy surrounding the current guidelines for the definition of non-curative resection.In addition,it emphasizes on the formulation of individualized management strategies for these patients in routine clinical practice.

Background Paraquat (PQ) is one of the most widely used herbicides in the world and a risk factor for Parkinson's disease (PD), but the mechanisms underlying PD are poorly understood. Single-cell RNA sequencing (scRNA-seq) technology can study cellular heterogeneity at genetic level, providing insights into the pathogenesis of PQ-induced PD. Objective To analyze the brain cell grouping of PQ-infected mice and the biological processes involved in the subpopulation of PD-like changes cells by scRNA-seq, and to provide clues for revealing potential mechanisms of PQ-induced PD-like changes in mouse brains. Methods Six male 6-week-old C57BL/6 mice were randomly divided into a control group and an experimental group, three mice in each group, and were intraperitoneally injected with 0 (saline) and 10.0 mg·kg⁻¹ PD respectively, once every two days, for 10 consecutive injections for modeling. After infection, mouse brains were taken and scRNA-seq was performed. Cell segmentation was performed according to gene expression characteristics of different cell types, PD-related cell subsets were screened by bioinformatics tools, and gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), gene set enrichment analysis (GSEA), protein interaction network analysis, and transcription factor prediction were performed on their characteristic genes. Finally, GO and KEGG analyses were performed on the differential genes of PD-associated cell subsets between the PQ-treated group and the control group, and the biological processes in which these genes may participate were analyzed. Results The sequencing data met quality control standards, a total of 55779 cells were obtained, and all cell dimensionality reduction analysis results showed that they could be further divided into 37 clusters, including 5 major cell types. Based on the KEGG analysis of the top 20 characteristic genes of each subpopulation, the specifically expressed Cluster 33 subpopulation (dopaminergic neurons) was screened and found to be significantly associated with PD. The results of GO analysis showed that the biological function of this subpopulation mainly enriched neurotransmitter transport and regulation. The results of GSEA analysis showed that the tyrosine metabolic pathway and the ligand-receptor interaction pathway of neural activity in brain tissues were significantly enriched. The analysis of transcriptional regulatory networks showed that 39 transcription factors were expressed differently. The metabolic pathway of the dopamine neuronal subset, endocytosis, Ras-associated protein 1 (Rap1) signaling pathway, and mitogen-activated protein kinase (MAPK) signaling pathway were all affected by PQ exposure, according to further analysis of its effects on this subpopulation. The GO analysis showed that differential genes were involved in biological processes such as ion transport and synaptic assembly regulation, and were involved in the cellular component formation of cytoplasm and synapses. Conclusion This study has initially mapped the transcriptome of single cells in the mouse brain after PQ exposure, and screened out the specific expression of Cluster 33 subgroup (dopaminergic neurons), which is significantly correlated with PD, and its biological function changes may be one of the mechanisms of PD-like changes in the mouse brain induced by PQ.

Cognitive impairment caused by chronic cerebral hypoperfusion (CCH) is associated with white matter injury (WMI), possibly through the alteration of autophagy. Here, the autophagy-lysosomal pathway (ALP) dysfunction in white matter (WM) and its relationship with cognitive impairment were investigated in rats subjected to two vessel occlusion (2VO). The results showed that cognitive impairment occurred by the 28th day after 2VO. Injury and autophagy activation of mature oligodendrocytes and neuronal axons sequentially occurred in WM by the 3rd day. By the 14th day, abnormal accumulation of autophagy substrate, lysosomal dysfunction, and the activation of mechanistic target of rapamycin (MTOR) pathway were observed in WM, paralleled with mature oligodendrocyte death. This indicates autophagy activation was followed by ALP dysfunction caused by autophagy inhibition or lysosomal dysfunction. To target the ALP dysfunction, enhanced autophagy by systemic rapamycin treatment or overexpression of Beclin1 (BECN1) in oligodendrocytes reduced mature oligodendrocyte death, and subsequently alleviated the WMI and cognitive impairment after CCH. These results reveal that early autophagy activation was followed by ALP dysfunction in WM after 2VO, which was associated with the aggravation of WMI and cognitive impairment. This study highlights that alleviating ALP dysfunction by enhancing oligodendrocyte autophagy has benefits for cognitive recovery after CCH.

Objective: To assess the anti-inflammatory and immunomodulatory activities of schisandrin B in mice. Methods: Kun-ming mice were randomly divided into 6 groups: the blank control group, the model control group, the positive control group( dexam-ethasone hydrochloride 5 mg·kg-1,levamisole hydrochloride 30 mg·kg-1), schisandrin B low (100 mg·kg-1), medium (200 mg ·kg-1) and high (400 mg·kg-1) dose groups. The inflammatory mice were induced by xylene and the immunosuppressed mice were induced by cyclophosphamide. The ear edema experiment, carbon clearance experiment and serum hemolysin experiment were per-formed with the swelling rate, swelling inhibition rate, carbon clearance index, phagocytic index, half hemolytic value, spleen index and thymus index as the investigation factors. Results: High dose and medium dose schisandrin B groups had significant inhibitory effect on mice ear edema when compared with the model control group(P<0. 05);There were significant differences between the high dose group and the positive group and there were significant differences in clearance index and phayxyitc index between high dose group and low dose group(P<0. 05), and high dose schisandrin B group had increased clearance index and phagocytic index in low immune function mice when compared with the model control group (P<0.05). Schisandrin B significantly increased HC50in low immune function mice when compared with the model control group (P<0. 01). Schisandrin B significantly increased the spleen index and thy-mus index in low immune function mice when compared with the model control group ( P<0. 01,and in a dose-dependent manner. Conclusion: Schisandrin B has significant anti-inflammatory and immune function enhancing effects.

Objective:To optimize the extraction process of Shenqi Lixin capsules through pharmacodynamic evaluation combined with orthogonal experiments with multi-index comprehensive evaluation.Methods:The congestive heart failure(CHF) model was established by intraperitoneal injection of doxorubicin in rats,and the LVEDD,LVESD,FS and LVEF of CHF rats were used as the indicators to screen the extraction process of the samples (process A was with the whole decoction of herbs and process B was with heterophylla powder mixed with the other herbs after boiling).The single factor tests and orthogonal tests were used to optimize the extraction process by taking the contents of astragaloside and tanshinone ⅡA and the quality of the decocted material as the indices,and adding water amount,decocting times and duration as the influencing factors.Results:Pharmacodynamic experiments indicated that the improvement effects of the samples from process B on cardiac symptoms and cardiac function in CHF rats were better than that of the samples from process A.The other medicinal materials were decocted by 12-fold amount of adding water,and repeated for 12 times with one hour for each time.The average extraction rate of astragaloside and tanshinone ⅡA was 61.82% and 50.07%,respectively,which was proven by the verification experiments.The average weight of the decoction was 6.02 g.Conclusion:The optimized extraction process of Shenq Lixin capsules is scientific,reasonable,stable and reliable.

Objective To analyze the reasons and treatment methods of high transprothetic pressure gradient after aortic valve replacement. Methods The clinical data of 45 patients with high transprothetic pressure gradient after aortic valve replacement were retrospectively analyzed. The patients were followed up for average 24.6 (12 - 40) months. The postoperative effective orifice area (EOA) of artificial valve was measured by transthoracic color Doppler ultrasound. Compared with published referred EOA of different artificial valve, there were 2 kinds results:measured EOA=referred EOA and measured EOA0.85 cm2/m2 and EOAI<0.85 cm2/m2. The reasons of high transprothetic pressure gradient were analyzed according to the above different standard. Results In the 45 patients with high transprothetic pressure gradient after aortic valve replacement, prosthesis-patient mismatch (PPM) was in 33 cases, and prosthetic dysfunction was in 10 cases, among whom 5 cases were because of thrombus (3 cases improved after increasing the dosage of warfarin, 2 cases underwent re-aortic valve replacement), 3 cases were because of severe bioprosthetic calcification (underwent re-aortic valve replacement), and 2 cases were because of prosthetic ring pannus and influenced movement of the leaflets (underwent re-aortic valve replacement). High flow in the left ventricular outflow tract occurred in 2 cases. The patients had no obvious discomfort, and did not receive special treatment. Four cases died, among whom 2 cases were because of severe PPM, and the other 2 cases were because of noncardiac. Conclusions Many reasons can result to the high transprothetic pressure gradient, and the PMM is the most common reason. Choosing the right treatment plan can improve the survival rate of patients.

BACKGROUNDCollaterals to occluded infarct-related coronary arteries (IRA) have been observed after the onset of acute ST-elevation myocardial infarction (STEMI). We sought to investigate the impact of early coronary collateralization, as evidenced by angiography, on myocardial reperfusion and outcomes after primary percutaneous coronary intervention (PCI).

METHODSAcute procedural results, ST-segment resolution (STR), enzymatic infarct size, echocardiographic left ventricular function, and major adverse cardiac events (MACE) at 6-month follow-up were assessed in 389 patients with STEMI undergoing primary PCI for occluded IRA (TIMI flow grade 0 or 1) within 12 hours of symptom-onset. Angiographic coronary collateralization to the occluded IRA at first contrast injection was graded according to the Rentrop scoring system.

RESULTSLow (Rentrop score of 0 or 1) and high (Rentrop score of 2 or 3) coronary collateralization was detected in 329 and 60 patients, respectively. Patients with high collateralization more commonly had prior stable angina and right coronary artery occlusion, but less often had left anterior descending artery occlusion. At baseline, these patients presented with less extent of ST-segment elevation and lower serum levels of creatine kinase myocardial band (CK-MB) and cardiac troponin I (cTnI). Procedural success rate, STR, corrected TIMI flame count, and area under the curve of CK-MB and cTnI measurements after the procedure were similar between patients with high collateralization and those with low collateralization (for all comparisons P > 0.05). There were no differences in left ventricular ejection fraction and rates of MACE at 6 months according to baseline angiographic collaterals to occluded IRA.

CONCLUSIONSIn patients with acute STEMI undergoing primary PCI within 12 hours of symptom-onset, coronary collateralization to the occluded IRA was influenced by clinical and angiographic features. Early recruitment of collaterals limits infarct size at baseline, but has no significant impact on myocardial reperfusion after the procedure and subsequent left ventricular function and clinical outcomes.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Angioplasty, Balloon, Coronary ; Coronary Angiography ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; diagnostic imaging ; therapy ; Treatment Outcome ; Young Adult

Objective To evaluate the influence of the gene polymorphisms of matrix metalloproteinase(mmp)-1 ,-2,-3 and -9 on coronary atherosclerotic plaque progression. Methods During the period of January 2005-December 2008, 80 patients with coronary heart disease underwent two times coronary angiography at authors' hospital. Based on the angiographic findings, the patients were classified into plaque progression group (n = 31 ) and plaque non-progression group (n = 49). Coronary atheroselerotic plaque progression was arbitrarily defined as that the minimal lumen diameter (MLD) of coronary artery showed a decrease ≥ 0.4 mm on the second coronary angiography. The detailed history and clinical examination results were collected, including serum concentrations of lipid profiling, fasting glucose and hs-CRP. Genotypings for polymorphic variances of MMP-1 (-1607 G/GG), MMP-2 (-955 A/C), MMP-3 (-1612 5A/6A ) and MMP-9 (-1562 C/T) were performed by polymerase chain reaction (PCR) and sequencing analysis in two groups.Comparison of the clinical characteristics and polymorphisms between two groups was made to assess their effects on coronary atherosclerotic plaque progression. Results More female patients and patients with acute coronary syndrome (ACS) were noted in patients with plaque progression compare to those with no progression (41.9% vs. 18.4%, P < 0.05 and 77.4% vs. 46.3%, P < 0.01, respectively).The serum hs-CRP level also significantly increased in group with plaque progression (0.26 ± 0.44 mg/L vs.0.02 ± 0.14 mg/L, P < 0.01). Multivariable logistic regression analysis revealed that serum hs-CRP concentration and ACS were independent risk factors of coronary atherosclerotic plaque progression (OR:12.63,95% CI:1.45-110.29, P < 0.05 and OR:2.99,95% CI:1.04-8.63, P < 0.05, respectively). The frequencies of 6A/6A genotype and 6A allele of MMP-3 promoter at location -1612 were significantly higher in group with plaque progression than that in group with no progression (87.1% vs. 53.1%, P < 0.01 and 93.5% vs. 75.5%, P < 0.01, respectively). However, no significant differences in the distribution of MMP-1,-2 and -9 polymorphisms existed between two groups. Conclusion ACS, feminine gender, high serum hs-CRP concentration and 5A/6A polymorphism in the MMP-3 gene promoter are closely associated with coronary atherosclerotic plaque progression. In addition, 5A/6A polymorphism of MMP-3 can be used as a marker for plaque progression.