Amyotrophic lateral sclerosis (ALS) is an irreversible, fatal neurodegenerative disorder whose incidence is positively correlated with the aging population. ALS is characterized by the progressive loss of motor neurons, leading to muscle weakness, atrophy, and ultimately respiratory failure. The pathogenesis of ALS involves multiple factors, including genetic and environmental influences, with genetic factors playing a particularly significant role. To date, several causative genes have been identified in ALS, such as the Cu/Zn superoxide dismutase 1 (Cu/Zn SOD1, also known as SOD1) gene, transactive response DNA-binding protein 43 (TDP-43) gene, fused in sarcoma (FUS) gene, and chromosome open reading frame 72 (C9orf72). Mutations in these genes have been found not only in familial ALS but also in sporadic ALS. Based on the identified ALS risk genes, various ALS animal models have been established through multiple approaches, including transgenic models, gene knockout/knock-in models, and adeno-associated virus-mediated overexpression models. These models simulate some typical pathological features of human ALS, such as motor neuron loss, ubiquitinated inclusions, and neuromuscular junction degeneration. However, these models still have limitations: (1) single-gene mutation models are insufficient to fully replicate the complex multi-factorial pathogenesis of sporadic ALS; (2) significant differences in microenvironmental regulation mechanisms and the rate of neurodegeneration between model organisms and humans may affect the accurate reproduction of disease phenotypes and the reliable evaluation of drug efficacy. To better understand the pathogenesis of ALS and promote the development of effective therapies, constructing and optimizing ALS animal models is crucial. This review aims to summarize commonly used ALS gene mutation mouse models, analyze their phenotypes and pathological characteristics, including transgenic mouse models, gene knockout/knock-in mouse models, and adeno-associated virus-mediated overexpression mouse models, and further discuss their specific applications in ALS pathogenesis research and drug development by comparing the advantages and limitations of each model.

Amyotrophic lateral sclerosis (ALS) is an irreversible, fatal neurodegenerative disorder whose incidence is positively correlated with the aging population. ALS is characterized by the progressive loss of motor neurons, leading to muscle weakness, atrophy, and ultimately respiratory failure. The pathogenesis of ALS involves multiple factors, including genetic and environmental influences, with genetic factors playing a particularly significant role. To date, several causative genes have been identified in ALS, such as the Cu/Zn superoxide dismutase 1 (Cu/Zn SOD1, also known as SOD1) gene, transactive response DNA-binding protein 43 (TDP-43) gene, fused in sarcoma (FUS) gene, and chromosome open reading frame 72 (C9orf72). Mutations in these genes have been found not only in familial ALS but also in sporadic ALS. Based on the identified ALS risk genes, various ALS animal models have been established through multiple approaches, including transgenic models, gene knockout/knock-in models, and adeno-associated virus-mediated overexpression models. These models simulate some typical pathological features of human ALS, such as motor neuron loss, ubiquitinated inclusions, and neuromuscular junction degeneration. However, these models still have limitations: (1) single-gene mutation models are insufficient to fully replicate the complex multi-factorial pathogenesis of sporadic ALS; (2) significant differences in microenvironmental regulation mechanisms and the rate of neurodegeneration between model organisms and humans may affect the accurate reproduction of disease phenotypes and the reliable evaluation of drug efficacy. To better understand the pathogenesis of ALS and promote the development of effective therapies, constructing and optimizing ALS animal models is crucial. This review aims to summarize commonly used ALS gene mutation mouse models, analyze their phenotypes and pathological characteristics, including transgenic mouse models, gene knockout/knock-in mouse models, and adeno-associated virus-mediated overexpression mouse models, and further discuss their specific applications in ALS pathogenesis research and drug development by comparing the advantages and limitations of each model.

Gentianopsis barbata (G. barbata) represents a significant plant species with considerable ornamental and medicinal value in China. This investigation sought to elucidate the primary constituents within the plant and investigate their pharmacological properties. Fifty triterpenoids (1-50), including nine previously undescribed compounds (1, 2, 7, 10, 20, 28, 29, 37, and 41) were isolated and characterized from the whole plants of G. barbata. Notably, compounds 1 and 2 exhibited the novel 3,4;9,10-diseco-24-homo-cycloartane triterpenoid skeleton. The isolated triterpenoids demonstrated substantial anti-inflammatory activity through inhibition of tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) cytokine secretion in LPS-induced RAW264.7 macrophages, and hepatoprotective effects by preventing tert-butyl hydroperoxide (t-BHP)-induced oxidative injury in HepG2 cells. These results demonstrate both the presence of diverse triterpenoids in G. barbata and their therapeutic potential for inflammatory and hepatic conditions, providing scientific evidence supporting the clinical application of this traditional Mongolian medicinal plant.
Triterpenes/isolation & purification* ; Mice ; Anti-Inflammatory Agents/isolation & purification* ; Animals ; Humans ; RAW 264.7 Cells ; Hep G2 Cells ; Interleukin-6/genetics* ; Tumor Necrosis Factor-alpha/genetics* ; Medicine, Mongolian Traditional ; Macrophages/immunology* ; Protective Agents/isolation & purification* ; Liver/drug effects* ; Gentianaceae/chemistry* ; Plant Extracts/chemistry* ; Molecular Structure

Aristolochic acid (AA), extracted from Aristolochiaceae plants, plays an essential role in traditional herbal medicines and is used for different diseases. However, AA has been found to be nephrotoxic and is known to cause aristolochic acid nephropathy (AAN).AA-induced acute kidney injury (AKI) is a syndrome in AAN with a high morbidity that manifests mitochondrial damage as a key part of its pathological progression. Melatonin primarily serves as a mitochondria-targeted antioxidant. However, its mitochondrial protective role in AA-induced AKI is barely reported. In this study, mice were administrated 2.5 mg/kg AA to induce AKI. Melatonin reduced the increase in Upro and Scr and attenuated the necrosis and atrophy of renal proximal tubules in mice exposed to AA. Melatonin suppressed ROS generation, MDA levels and iNOS expression and increased SOD activities in vivo and in vitro. Intriguingly, the in vivo study revealed that melatonin decreased mitochondrial fragmentation in renal proximal tubular cells and increased ATP levels in kidney tissues in response to AA. In vitro, melatonin restored the mitochondrial membrane potential (MMP) in NRK-52E and HK-2 cells and led to an elevation in ATP levels. Confocal immunofluorescence data showed that puncta containing Mito-tracker and GFP-LC3A/B were reduced, thereby impeding the mitophagy of tubular epithelial cells. Furthermore, melatonin decreased LC3A/B-II expression and increased p62 expression. The apoptosis of tubular epithelial cells induced by AA was decreased. Therefore, our findings revealed that melatonin could prevent AA-induced AKI by attenuating mitochondrial damage, which may provide a potential therapeutic method for renal AA toxicity.

Objective:To investigate the genomic manifestation and pathogenesis of osteosarcoma with different relapse pattens, which were respectively initially presented with bone metastasis or pulmonary metastasis.Methods:From May 1, 2021 to October 1, 2021, 38 fresh tumor specimens and some paraffin-embedded specimens of high-grade osteosarcoma were collected in Peking University People's Hospital, including 29 males and 9 females, aged 19.6±2.2 years (range, 6-61 years). Among the 38 cases, 12 cases had initial bone metastasis (group A) and 26 cases had initial lung metastasis (group B), of which 15 cases (40%, 15/38) had paired specimens of primary and metastatic lesions. Based on Illumina NovaSeq 6000, we analyzed whole-exome sequencing (WES) as well as transcriptome for osteosarcoma with paired samples in different relapse patterns. During all their treatment courses, we also collected their paired samples to reveal these tumors' evolution. We sought to redefine disease subclassifications for osteosarcoma based on genetic alterations and correlate these genetic profiles with clinical treatment courses to elucidate potential evolving cladograms.Results:We found that osteosarcoma in group A mainly carried single-nucleotide variations (83%, 10/12), displaying higher tumor mutation burden [4.9 (2.8, 12.0) & 2.4 (1.4, 4.5), P=0.010] and neoantigen load [743.0 (316.5, 1,034.5) & 128.5 (49.0, 200.5), P=0.003], while those in group B mainly exhibit structural variants (58%, 15/26). The mutation spectrum showed that there was a significant difference in age-related gene imprinting 1 between the bone metastasis group and the lung metastasis group ( P=0.005). Samples were randomly selected from group A (3 patients) to investigate immunologic landscape by multiplex immunohistochemistry, from which we noticed tertiary lymphatic structure from one patient from group A. High conservation of reported genetic sequencing over time was found in their evolving cladograms. Conclusion:Osteosarcoma with mainly single-nucleotide variations other than structural variants might exhibit biological behavior predisposing toward bone metastases with older in age as well as better immunogenicity in tumor microenvironment.

Objective:To observe the efficacy and safety of urokinase arterial thrombolysis in the treatment of central retinal artery occlusion (CRAO) at different time window.Methods:A retrospective study. From January 2014 to November 2019, 157 eyes (157 CRAO patients) in the Xi’an People's Hospital (Xi’an Fourth Hospital) were included in the study. There were 120 males and 37 females, with the average age of 54.87±12.12 years. The mean onset time was 65.66±67.44 h. All patients were tested with BCVA using international standard visual acuity chart, and the results were converted into logMAR visual acuity record. The arm-retinal circulation time (A-Rct) and the filling time (FT) of retinal arterial trunk-terminal filling time were measured by FFA. The mean logMAR BCVA was 2.44±0.46, the mean A-Rct and FT were 27.72±9.78 and 13.58±14.92 s respectively. According to the time window, the patients were divided into the onset 3-72 h group and the onset 73-240 h group, which were 115 patients and 42 patients respectively. There were no statistically significant difference between the 3-72 h group and the 73-240 h group in age, A-Rct and LogMR BCVA before treatment ( χ2=-0.197, -1.242, -8.990; P=0.844, 0.369, 0.369); the difference was statistically significant in FT comparison ( χ2=-3.652, P=0.000). Urokinase artery thrombolytic therapy was performed at different time window of 3-24 h, 25-72 h, 73-96 h, 97-120 h, 121-240 h after the onset of onset. Age and A-Rct of patients with different treatment time windows were compared, and the differences were not statistically significant ( χ2=6.588, 6.679; P=0.253, 0.246).In comparison of FT and logMAR BCVA, the difference was statistically significant ( χ2 =30.150, 71.378; P=0.000, 0.000). FFA was rechecked 24 hours after treatment, BCVA was rechecked 30 days after treatment. The changes of A-Rct, FT and BCVA before and after treatment were compared and analyzed. The occurrence of adverse reactions during and after treatment were observed. The two groups of measurement data were compared. The t test was used for those with normal distribution and χ2 test was used for those with non-normal distribution. Spearman correlation analysis was used to analyze the correlation between onset time and the difference of A-Rct, FT shortening time and logMAR BCVA after treatment. Results:At 24 h after CRAO treatment, A-Rct and FT of 157 cases were 19.64±6.50 and 6.48±7.36 s respectively, which were significantly shorter than those before treatment, and the differences were statistically significant ( χ2=-16.236, -14.703; P=0.000, 0.000). The logMAR BCVA at 30 d after treatment was 1.72±0.76, which was significantly higher than that before treatment. The difference was statistically significant ( χ2=-14.460, P=0.000). After CRAO urokinase arterial thrombolysis at different time window, there were statistically significant differences in A-Rct shortening time, FT shortening time, and logMAR BCVA difference ( χ2=12.408, 24.200, 104.388; P=0.030, 0.000, 0.000). There was no statistically significant difference between the 3-72 h group and the 73-240 h group ( χ2 =-1.042, P=0.297) in shortening time of A-Rct after treatment. The difference of FT shortening time was statistically significant ( χ2=-3.581, P=0.000). The difference of logMAR BCVA was statistically significant ( χ2=-9.905, P=0.000). The results of Spearman correlation analysis showed that there was no correlation between the onset time and the shortening time of A-Rct and FT after treatment ( rp=-0.040, -0.081; P=0.436, 0.115), and negative correlation with the logMAR BCVA difference ( rp=-0.486, P=0.000). One case of intracranial hemorrhage occurred after treatment, and it improved after dehydration to reduce cerebral edema, scavenging free radicals and brain protection. Conclusions:Urokinase arterial thrombolytic therapy is effective for CRAO within time window of 3-240 h, A-Rct, FT and LogMRA BCVA are all improved. However, with the prolongation of thrombolytic therapy time window, the therapeutic effect of urokinase arterial thrombolytic therapy is decreased. The therapeutic effect of Urokinase arterial thrombolytic therapy was better within 72 h.

OBJECTIVETo determine the frequency of spinocerebellar ataxia type 31 (SCA31) related mutations among patients from mainland China.

METHODSFor a cohort of molecularly unassigned patients comprised of 295 SCA patients (including 98 probands from families featuring autosomal dominant SCA and 197 sporadic cases) and 81 patients with hereditary spastic paraplegia (HSP) (including 23 probands from families with autosomal dominant HSP and 58 sporadic cases),TGGAA pentanucleotide expansion insertional mutation of the BEAN/TK2 gene was detected using repeat-primed PCR followed by capillary gel electrophoresis.

RESULTSNo TGGAA pentanucleotide insertion expansion in BEAN/TK2 gene was identified in the above cohort.

CONCLUSIONSCA31 is an extremely rare subtype of SCA and should not be included in routine genetic screening in mainland China.

Objective To explore the diagnosis and treatment of poorly differentiated endocardial sarcoma. Method The clinical data of a child with poorly differentiated endocardial sarcoma was retrospectively analyzed. Results One-year-old girl was admitted for diarrhea, polypnea, cyanosis, and cough. Abnormal heart sound was found by auscultation. Leads Ⅱ, Ⅲ, and aVF of ECG showed high peaked P wave. The diagnosis of poorly differentiated endocardial sarcoma was confirmed by echocardiography and pathology after cardiac operation. Three months after discharge from the hospital, the patient suddenly came into coma and died. Conclusion The diagnosis of poorly differentiated endocardial sarcoma is mainly based on clinical manifestations, echocardiography and pathology. Surgical resection is the first choice and chemotherapy and radiotherapy play a supporting role. However, there is no cure for it currently.

To describe a novel particles surface display system which is consisted of gram-positive enhancer matrix (GEM) particles and anchor proteins for bacteria-like particles vaccines, we treated Lactobacillus rhamnosus GG bacteria with 10% heated-TCA for preparing GEM particles, and then identified the harvested GEM particles by electron microscopy, RT-PCR and SDS-PAGE. Meanwhile, Escherichia coli was induced to express hybrid proteins PA3-EGFP and P60-EGFP, and GEM particles were incubated with them. Then binding of anchor proteins were determined by Western blotting, transmission electron microscopy, fluorescence microscopy and spectrofluorometry. GEM particles preserved original size and shape, and proteins and DNA contents of GEM particles were released substantially. The two anchor proteins both had efficiently immobilized on the surface of GEM. GEM particles that were bounded by anchor proteins were brushy. The fluorescence of GEM particles anchoring PA3 was slightly brighter than P60, but the difference was not significant (P>0.05). GEM particles prepared from L. rhamnosus GG have a good binding efficiency with anchor proteins PA3-EGFP and P60-EGFP. Therefore, this novel foreign protein surface display system could be used for bacteria-like particle vaccines.

Objectives To evaluate the effects of "disease-characteristics nursing quality improvement" program(DNQIP)combined with targeted monitoring in reducing the incidence of ventilator associated pneumonia (VAP)among patients with mechanical ventilation and to provide evidence for the prevention of VAP. Methods This was a prospective intervention study. All mechanical ventilation patients admitted in stroke intensive care unit(SICU)of our hospital from 2007 to 2015 were selected as the research object. Targeted monitoring was implemented in patients admitted from 2007 to 2011,and DNQIP was formed based on the monitoring results. DNQIP combined with targeted monitoring was implemented from 2012 to 2013 as the first phase,and the second phase was from 2014 to 2015.Results From 2007 to 2015,the number of new patients per year and the total time of using breathing machine in SICU presented a tendency of increasing while the number of patients with VAP and the incidence of VAP presented a tendency of decreasing year by year. The implementation of DNQIP in the first phase from 2012 to 2013(incidence of VAP was 5.4 ‰),compared with the data before implementation from 2007 to 2011(incidence of VAP was 13.8‰),the incidence of VAP was significantly reduced(χ2=9.029,P=0.003). The incidence of VAP(5.3‰)in the second phase from 2014 to 2015,was significantly reduced compared with the incidence of VAP(13.8‰)from 2007 to 2011(χ2=9.559, P=0.002).Conclusions The implementation of targeted monitoring is helpful in finding problems existing in the prevention of VAP. DNQIP in patients with mechanical ventilation can effectively reduce the incidence of VAP and can refine professional assessment and health education. It is also good for the standardization and evidence-based care in the whole process provided for patients. DNQIP combined with targeted monitoring can realize the continuous quality improvement on the basis of evidence-based theory.