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.

Objective To investigate the correlation between novel bone metabolism markers and the degree of aortic calcification as well as cardiovascular event risk in elderly patients treated by PD.Methods A prospective trial was conducted on 230 elderly patients receiving continuous am-bulatory PD in our department from February 2022 to February 2024.According to the occur-rence of cardiovascular events during dialysis treatment,they were divided into a cardiovascular event group(n=92)and a control group(n=138).Relevant clinical data were collected,aortic calcification was assessed using AAC scores,and serum levels of bone metabolism markers,inclu-ding osteoprotegerin,TRACP,and PINP were measured.Results The serum levels of osteoprote-gerin,TRACP and PINP were significantly higher in the cardiovascular event group than the con-trol group(P＜0.01).The cardiovascular event group had obviously severe calcification and higher AAC score than the control group(P＜0.01).The serum levels of the three bone metabolism markers were notably higher in the patients with severe calcification than those with moderate calcification,followed by mild calcification in turn(P＜0.01).Spearman correlation analysis indi-cated that the levels of the three indicators were positively correlated with the degree of aortic cal-cification in elderly PD patients(r=0.465,P=0.000;r=0.396,P=0.000;r=0.434,P=0.000).Multivariate logistic regression analysis showed that these three indicators were risk factors for cardiovascular events in elderly PD patients(P＜0.01).Conclusion The three bone metabolism markers are significantly correlated with aortic calcification severity and cardiovascular event risk in elderly PD patients.Monitoring these marker levels may be helpful for the assessment and man-agement of cardiovascular risk.

Objective:To analyze the clinical characteristics of children hospitalized with acute lower respiratory tract infections (ALRTI) caused by respiratory syncytial virus (RSV) in Kunming area in 2023.Methods:The clinical data of children with RSV-ALRTI admitted to Kunming Children's Hospital from January to December 2023 were retrospectively analyzed.According to the severity of the disease and whether the children developed severe pneumonia,they were divided into common group and severe group.The general data,clinical manifestations,laboratory indexes,imaging results,treatment and outcomes of the children between the two groups were compared.Multinomial Logistic regression analysis was used to study the risk factors for severe RSV-ALRTI.Results:A total of 696 children with RSV-ALRTI were included,including 432 males (62.1%) and 264 females (37.9%),with a median age of 1.30 (0.40,3.38) years.The onset of RSV-ALRTI occured throughout the year,with a peak incidence in May to August (60.6%).One hundred and seventy-one of 696 samples were tested RSV gene sequence,and two genotypes of RSV A-ON1(60.8%) and RSV B-BA9(39.2%) were detected.Among the 696 children,480 (69.0%) were in the common group and 216 (31.0%) were in the severe group.The age of the children in the severe group was lower than that of the common group,and the proportions of age <1 year,preterm birth,congenital diseases (congenital anomalies of airway development,congenital heart disease),combined Mycoplasma pneumoniae infections,and combined bacterial infections were higher than those of the common group ( P<0.05).There was no statistically significant differences in fever and cough symptoms between the two groups ( P>0.05),but the proportions of children in the severe group with wheezing,shortness of breath,dyspnea,vomiting,and diarrhea were significantly higher than those in the common group ( P<0.05),and the proportions of moist rales and wheezing heard by lung auscultation were significantly higher than those in the common group ( P<0.05).The levels of leukocytes,neutrophils,neutrophil percentage,platelets,procalcitonin,lactate dehydrogenase,alanine aminotransferase,aspartate aminotransferase,creatine kinase isoenzyme,and ferritin in the severe group were higher than those in the common group,while the levels of hemoglobin,lymphocyte percentage,albumin,creatinine,IgG,IgM,IgA,interleukin 6,D-dimer,and fibrin were lower than those in the common group ( P<0.05).The proportions of pulmonary consolidation and pleural effusion in the severe group were higher than those in the common group,and the severe group was more likely to have bilateral pulmonary consolidation ( P<0.05).Both groups of children were given interferon aerosol inhalation for antiviral treatment,and actively symptomatic and supportive treatment.One case in the severe group died during hospitalization,and the remaining 695 cases were cured and discharged.The proportions of children in the severe group using antibiotics,dual antibiotics,intravenous hormones,intravenous immunoglobulin,complications,transfer to PICU,use of invasive mechanical ventilation,length of hospital stay,and treatment costs were significantly higher than those in the common group ( P<0.001).Multinomial Logistic regression analysis showed that low age ( OR=0.878,95% CI 0.800~0.963, P=0.006),congenital diseases (congenital anomalies of airway development,congenital heart disease) ( OR=2.892,95% CI 1.977~4.233, P<0.001),and co-infection with bacteria ( OR=1.846,95% CI 1.268~2.686, P=0.001) were risk factors for the development of severe pneumonia in RSV-infected children. Conclusions:In 2023,RSV-ALRTI in children in Kunming area had an incidence throughout the year,and the peak age group of incidence was predominantly <1 year old,and the main genotype of infection was RSV A-ON1.Children with RSV-ALRTI who are young,suffering from congenital diseases (congenital anomalies of airway development,congenital heart disease) and complicated with bacterial infection are more likely to develop into severe diseases,and should be paid close attention and active intervention to reduce the incidence of critical illness.

Objective To verify the accuracy of a Non-Contact Sleep Monitoring Mattress(NCSMM)based on body movement during sleep in assessing sleep quality of patients before neurosurgery in order to provide a more portable and efficient assessment tool for clinical staff.Methods A single-arm trial was conducted on 114 inpatients admitted in our department selected with convenience sampling.Sleep quality data of 1 night were collected through 5 sleep quality assessment tools,including NCSMM,polysomnography(PSG),Patient-Reported Outcome Measurement Information System(PROMIS)Sleep Disturbance scale,Richards-Campbell Sleep Scale(RCSQ),and a wearable device(smart watch for body movements and sleep quality monitoring).The sleep efficiency(≤85%)obtained by PSG was used as the diagnostic standard for sleep disorders.The area under the receiver operating characteristic curve(AUC),sensitivity,specificity,positive predictive value,negative predictive value,and Youden index were calculated for the other 4 tools to evaluate and compare their diagnostic effectiveness.Results The AUC value for NCSMM,PROMIS,RCSQ and smart watch was 0.788(95%CI:0.687～0.888,P<0.001),0.664(95%CI:0.543～0.784,P=0.02),0.723(95%CI:0.600～0.846,P=0.001)and 0.750(95%CI:0.654～0.846,P<0.001),respectively.The diagnostic accuracy rate was 0.774,0.559,0.742 and 0.602,with corresponding Youden index value of 0.488,0.321,0.456,and 0.459.NCSMM demonstrated the best AUC value,sensitivity and Youden index when compared with the other 3 tools.Conclusion NCSMM shows high accuracy in assessing sleep quality in pre-neurosurgery inpatients,and it is a viable portable and efficient assessment tool in clinical practice.

Objective:To screen differentially expressed genes (DEGs) associated with liver cancer by bioinformatics analysis method, and to investigate the mechanism of the minichromosome maintenance protein 2 ( MCM2) and replication factor C subunit 4 ( RFC4) genes in liver cancer in vitro. Methods:Gene expression profiling data of 80 and 36 hepatocellular carcinoma tissues and 307 and 13 cirrhotic tissues were obtained from the GSE25097 and GSE98620 datasets of the gene expression analysis (GEO) database, respectively. Gene expression profiling data of 374 liver cancer tissues and 50 normal liver tissues were downloaded from the cancer genome atlas (TCGA) database. Limma and DESeqs R software were used to process the gene expression profiling data, construct protein-protein interaction networks, and analysis the relevance of these genes to survival. Weighted gene co-expression network analysis was performed to screen out the core genes. Liver cancer SMMC7721 cells were transfected with MCM2 blank plasmid (MCM2 control group), MCM2 overexpression plasmid [MCM2 WT1 group, MCM2 WT2 (2-fold WT1) group], RFC4 blank plasmid (RFC4 control group), and RFC4 overexpression plasmid [RFC4 WT1 group, RFC4 WT2 (2-fold WT1) group], respectively. The expression of MCM2 and RFC4 in liver cancer cell lines and their transfection levels were detected by real-time fluorescence quantitative reverse transcription PCR and Western blotting. The effects of MCM2 and RFC4 on the proliferation of hepatocellular carcinoma cells were detected by MTT assay and cell cloning assay, respectively. The effects of MCM2 and RFC4 on the migration of liver cancer cells were determined by the scratch assay. The effects of MCM2 and RFC4 on liver cancer cell invasion were detected by Transwell assay.Results:By bioinformatic analysis, 9 HCC DEGs were selected, including ubiquitin conjugating enzyme E2 T ( UBE2T), aurora kinase A ( AURKA), targeting protein for Xklp2 ( TPX2), MCM2, RFC4, ribonucleoside-diphosphate reductase subunit M2 ( RRM2), serine peptidase inhibitory factor Kazal type 1 ( SPINK1), collagen type XV alpha 1 chain ( COL15A1) and C-C motif chemokine 25 ( CCL25). Among the six genes associated with clinical stages, the MCM2 and RFC4 genes were found to be strongly associated with prognosis in liver cancer. The relative protein expression of MCM2 and RFC4 in HepG2 cells (1.83±0.07, 1.44±0.09) and SMMC7721 cells (1.74±0.05, 1.43±0.08) was higher than that in MIHA cells (1.00±0.02, 1.00±0.03), and all the differences were statistically significant (all P<0.05). The relative gene expression of MCM2 and RFC4 in HepG2 cells (14.30±0.12, 5.10±0.18) and SMMC7721 cells (10.60±0.11, 7.60±0.07) was higher than that in MIHA cells (1.00±0.05, 1.00±0.03), and all the differences were statistically significant (all P<0.05). Compared with the MCM2 control group, the absorbance values [(0.28±0.01 and 0.21±0.01) vs 0.18±0.03], the number of clonal cells [(717±12 and 782±29) cells vs (389±17) cells], the percentage migration [(0.43±0.02 and 0.68±0.01) vs 0.15±0.06], and the number of cellular invasions [(933±21 and 821±11) cells vs (409±16) cells] were higher in the MCM2 WT1 and MCM2 WT2 groups, and the differences were all statistically significant (all P<0.05). Compared with the RFC4 control group, the absorbance values [(0.30±0.02 and 0.21±0.01) vs 0.17±0.02], the number of cloned cells [(571±11 and 728±9) cells vs (373±23) cells], the percentage migration [(0.75±0.11 and 0.67±0.04) vs 0.34±0.07], and the number of cell invasion [(835±26 and 818±18) cells vs (629±12) cells] were higher in the RFC4 WT1 and the RFC4 WT2 groups, and the differences were statistically significant (all P<0.05). Conclusions:MCM2 and R FC4 genes play a role in promoting tumorigenesis and growth in hepatocellular carcinoma.

Objective To explore the long-term and short-term therapeutic effects and mechanisms of electroacupuncture at Baihui(GV20)point and Dazhui(GV14)point in rats with ischemic stroke.Methods Thirty rats were randomly divided into a sham-operated group,a model group,and an electroacupuncture group,with 10 rats in each group.The model group and the electroacupuncture group were subjected to middle cerebral artery occlusion(MCAO)using the modified Longa method.The electroacupuncture group received electroacupuncture at Baihui point and Dazhui point once daily for one week.Long-term neurological recovery was assessed using the adhesive removal test and the forelimb placement test at 14,21 and 28 days after MCAO.On the second day after MCAO,cerebral cortical histopathological changes were observed using transmission electron microscopy,the expression of Iba-1 in cortical tissue was detected by immunofluorescence,and the expression levels of hypoxia-inducible factor 1α(HIF-1α),vascular endothelial growth factor(VEGF),interleukin 1β(IL-1β),and tumor necrosis factor α(TNF-α)in cortical tissue were measured by Western Blot.Results(1)Compared with the sham-operated group at the same time points,the model group showed prolonged adhesive removal time and increased forelimb stepping times at 14,21 and 28 days after MCAO(P＜0.05).Compared with the model group at the same time points,the electroacupuncture group exhibited significantly shortened adhesive removal time and reduced forelimb stepping times at 14,21 and 28 days after MCAO(P＜0.05).Within 28 days after MCAO,the adhesive removal time was gradually decreased,and forelimb stepping times were gradually reduced.(2)On the second day after MCAO:compared with the sham-operated group,the model group showed degeneration and necrosis of microglia in cortical ischemic penumbra,mitochondrial damage,increased expression of Iba-1 in cortical tissue,and elevated levels of HIF-1α,VEGF,IL-1β and TNF-α,with statistically significant differences(P＜0.05).Compared with the model group,the electroacupuncture group exhibited the improvement of microglia injury in cortical ischemic penumbra,reduced expression of Iba-1 in cortical tissue and decreased levels of HIF-1α,VEGF,IL-1β and TNF-α,with statistically significant differences(P＜0.05).Conclusion Electroacupuncture at Baihui point and Dazhui point can improve symptoms in rats with ischemic stroke in the long term and ameliorate degeneration and necrosis of cortical microglia in the ischemic penumbra in the short term.The mechanism may be related to the downregulation of HIF-1α,VEGF,IL-1β and TNF-α expressions in cerebral cortical tissue and the inhibition of microglial transformation to the M1 phenotype.

Objective To compare the effects of Tripterygium wilfordii polyglycosides,cyclophosphamide,and cisplatin on the establishment of a mouse model of diminished ovarian reserve(DOR).Methods Mice were randomly divided into the following treatment groups:control(Ctrl),Tripterygium wilfordii polyglycosides(TWP),cyclophosphamide(CTX),and cisplatin(DDP).Mice in the TWP group received a 50 mg/kg suspension of Tripterygium wilfordii polyglycosides by gavage for 14 days,mice in the CTX group received a 20 mg/kg cyclophosphamide suspension by intraperitoneal injection for 14 days,and mice in the DDP group received a 1.5 mg/kg cisplatin solution by intraperitoneal injection for 14 days.The body weight,uterine index,and ovarian index were recorded,the estrous cycle was monitored using the vaginal smear method,and the levels of anti-Mullerian hormone(AMH),estradiol(E2),follicle stimulating hormone(FSH),and luteinizing hormone(LH)were detected using ELISA.Hematoxylin and eosin staining was used to detect ovarian follicle development.The rates of oocyte maturation and fertility were analyzed.Results The three treatment groups of mice all showed the following:significantly decreased body weight and ovarian index(P＜0.05);apparent disorder of the estrous cycle;significantly decreased levels of AMH and E2(P＜0.05);decreased and increased rates of developing follicles and atretic follicles,respectively(P＜0.05);and significantly decreased rates of oocyte maturation,pregnancy,and live birth(P＜0.05).Conclusions DOR mouse models were successfully constructed using Tripterygium wilfordii polyglycosides,cyclophosphamide,or cisplatin,as evidenced by decreased body weight and ovarian index,disordered estrous cycle and hormones,and DOR function,resulting in reduced rates of oocyte maturation,pregnancy,and total number of live births.These DOR effects were most appropriate in the cyclophosphamide group.

Objective:To investigate the prevalence of flight fatigue among helicopter flying personnel and analyze its contributors in order to provide data for related interventions.Methods:A cross-sectional study was conducted among 404 helicopter flying personnel between October 8, 2021 and July 31, 2022. Data was collected using a self-designed questionnaire, involving the demography of these subjects, sleep-related factors, flight fatigue, perceived causes of fatigue and coping strategies. The Pittsburgh Sleep Quality Index (PSQI), National Aeronautics and Space Administration Task Load Index (NASA-TLX) and Modified Fatigue Impact Scale (MFI-20) were used to assess sleep quality, mental workload, and levels of flight fatigue over the past month. The total scores of MFI-20 were compared across demographic groups, and correlations with PSQI and NASA-TLX scores were analyzed. Multiple linear regression was performed to identify the determinants of flight fatigue.Results:①Demography: among the 404 helicopter flying personnel, 92.8% (375/404) were pilots and 7.2% (29/404) navigators. As for years of service, 41.6% (168/404) served less than 5 years, while 58.4% (236/404) served more than 5 years. 37.9% (153/404) had a family history of insomnia. 18.8% (76/404) did not habitually nap, 68.9% (226/328) napped for ≤30 min, 31.1% (102/328) napped over 30 min, and 18.3% (74/404) had insomnia over the past month. As for helicopter flying personnel, 75.5% (305/404) reported experiencing fatigue, with 69.1% (279/404) attributing it to flight-related factors and 51.5% (208/404) using coffee as a countermeasure.②Scale scores: the total score of PSQI was [5 (3, 7)], while the highest daytime dysfunction score was [1(0, 2)]. The total score of NASA-TLX was [39.19 (26.57, 51.97)], and the effort score was the highest [10.31(5.07, 14.60)]. The total score of MFI-20 averaged (47.28±14.88), with the mental fatigue score being the highest [(10.03±4.42)]. ③Comparisons of MFI-20 total scores: flying personnel with ≤5 years of flying experience had higher MFI-20 total scores than those with >5 years, and those with a family history of insomnia had higher scores than those without ( t=3.35, 2.44, P=0.001, 0.015). Individuals with insomnia over the past month had higher scores than non-insomniacs ( t=3.33, P=0.001). Significant differences in MFI-20 scores were observed based on nap duration ( F=19.95, P<0.001). Non-nappers had higher scores than those napping for ≤30 min ( P=0.005). Flying personnel who napped for >30 min had higher scores than those did not ( P=0.043) or napped for ≤30 min ( P<0.001). ④Correlation analysis: the total score of MFI-20 was positively correlated with sleep quality, sleep latency, sleep disturbances, hypnotic medications, daytime dysfunction, and the total score of PSQI ( r=0.118-0.226, all P<0.05), but negatively with sleep duration ( r=-0.136, P=0.006). The total score of MFI-20 was positively correlated with mental demand, physical demand, and the total score of NASA-TLX ( r=0.119, 0.168, 0.184, P=0.017, 0.001, <0.001). ⑤Multiple linear regression analysis: the determinants of flight fatigue included aircraft types ( B=-4.956, 95% CI:-8.124--1.788), nap duration ( B=3.693, 95% CI: 1.267-6.119), sleep latency ( B=2.371, 95% CI: 0.229-4.513), sleep duration ( B=-7.383, 95% CI:-10.008--4.758), daytime dysfunction ( B=5.003, 95% CI: 2.967-7.039) and physical workload ( B=0.611, 95% CI: 0.324-0.898). Conclusions:Helicopter flying personnel are vulnerable to flight fatigue, which is strongly linked to sleep quality and mental workload. It is crucial to address flying personnel′s self-perceived fatigue, care about fatigue manifestations across aircraft types, and implement targeted interventions to improve sleep quality and reduce mental workload.

This article combined the current status of the industry and standards for pharmaceutical composite films and bags,and summarized the relevant content of the guiding principles for the Pharmacopoeia of the People's Republic of China(2025 Edition)on pharmaceutical composite films and bags.It analyzed the production requirements,usage requirements,key quality attributes,and control requirements improvement details,providing reference for assisting relevant parties to understand and use the pharmacopoeia guiding principles,and helping to improve the quality control level of the industry.