1.Establishment of a Rat Model of Alzheimer's Disease by Introducing Human Triple Mutant APP Gene into Hippocampus via Brain Stereotactic Technology
Linlin XIAO ; Yixuan YANG ; Shanshan LI ; Lanshiyu LUO ; Siwei YIN ; Juming SUN ; Wei SHI ; Yiqiang OUYANG ; Xiyi LI
Laboratory Animal and Comparative Medicine 2025;45(3):269-278
Objective To establish a rat model of Alzheimer's disease (AD) expressing human triple mutant amyloid precursor protein (APP) in the hippocampus, and to provide a model for the study of disease mechanisms and drug development. Methods Twenty-four 12-week-old SPF-grade female SD rats were randomly divided into a blank control group, a virus control group and an experimental group, with eight rats in each group; among them, the experimental group received a stereotaxic injection of adeno-associated virus (AAV) carrying the human triple mutant APP and NanoLuc luciferase genes into the hippocampus. In vivo imaging was used to observe viral expression in the brains of rats in each group, the novel object recognition test was used to assess the recognition memory of the rats in each group, real-time fluorescent quantitative PCR was used to detect the expression level of the APP gene, HE staining was used to examine the brain histopathology, Nissl staining was used to assess the hippocampal lesions, and immunohistochemistry was used to detect the deposition of amyloid β-protein (Aβ). Results In vivo imaging showed that reporter fluorescence was detected in the brains of rats in both experimental and virus control groups. Fluorescence quantitative PCR showed that the expression level of the APP gene was significantly increased in the brains of rats in the experimental group (P<0.01). Novel object recognition test revealed that the recognition memory of rats in the experimental group was significantly reduced compared with that of the blank control group (P<0.01). Six months after recombinant AAV virus infection, HE staining and Nissl staining of brain tissues showed that the number of neurons and Nissl bodies in the CA1 region of the hippocampus in the experimental group was reduced and disorganized; immuno-histochemistry testing of the CA1 region of the hippocampus and the pyramidal cell layer of the experimental group revealed prominent brown deposits, indicating Aβ protein deposition. Conclusion The rat model successfully established by stereotaxic injection and AAV-mediated delivery of human triple mutant APP gene exhibits typical AD features, providing a valuable animal model for studying AD pathology and developing drug therapies targeting Aβ protein deposition.
2.Establishment of a Rat Model of Alzheimer's Disease by Introducing Human Triple Mutant APP Gene into Hippocampus via Brain Stereotactic Technology
Linlin XIAO ; Yixuan YANG ; Shanshan LI ; Lanshiyu LUO ; Siwei YIN ; Juming SUN ; Wei SHI ; Yiqiang OUYANG ; Xiyi LI
Laboratory Animal and Comparative Medicine 2025;45(3):269-278
Objective To establish a rat model of Alzheimer's disease (AD) expressing human triple mutant amyloid precursor protein (APP) in the hippocampus, and to provide a model for the study of disease mechanisms and drug development. Methods Twenty-four 12-week-old SPF-grade female SD rats were randomly divided into a blank control group, a virus control group and an experimental group, with eight rats in each group; among them, the experimental group received a stereotaxic injection of adeno-associated virus (AAV) carrying the human triple mutant APP and NanoLuc luciferase genes into the hippocampus. In vivo imaging was used to observe viral expression in the brains of rats in each group, the novel object recognition test was used to assess the recognition memory of the rats in each group, real-time fluorescent quantitative PCR was used to detect the expression level of the APP gene, HE staining was used to examine the brain histopathology, Nissl staining was used to assess the hippocampal lesions, and immunohistochemistry was used to detect the deposition of amyloid β-protein (Aβ). Results In vivo imaging showed that reporter fluorescence was detected in the brains of rats in both experimental and virus control groups. Fluorescence quantitative PCR showed that the expression level of the APP gene was significantly increased in the brains of rats in the experimental group (P<0.01). Novel object recognition test revealed that the recognition memory of rats in the experimental group was significantly reduced compared with that of the blank control group (P<0.01). Six months after recombinant AAV virus infection, HE staining and Nissl staining of brain tissues showed that the number of neurons and Nissl bodies in the CA1 region of the hippocampus in the experimental group was reduced and disorganized; immuno-histochemistry testing of the CA1 region of the hippocampus and the pyramidal cell layer of the experimental group revealed prominent brown deposits, indicating Aβ protein deposition. Conclusion The rat model successfully established by stereotaxic injection and AAV-mediated delivery of human triple mutant APP gene exhibits typical AD features, providing a valuable animal model for studying AD pathology and developing drug therapies targeting Aβ protein deposition.
3.Application of speech repetition task in the detection of mild cognitive impairment
Xiaoxiao YIN ; Siwen WANG ; He WANG ; Linlin GAO ; Zhi REN ; Qinwen WANG
Chinese Journal of Nervous and Mental Diseases 2024;50(4):247-251
Mild cognitive impairment(MCI)is often regarded as a prodromal stage of dementia,primarily characterized by mild decline in cognitive function.Due to the mild nature of its symptoms,many MCI cases miss the opportunity for intervention.Research indicates that linguistic changes in MCI may precede other cognitive symptoms,providing an opportunity for early identification and intervention.These linguistic changes include abnormalities in speech rate,pronunciation,and intonation.Speech repetition tasks,such as the five-word test,delayed digit matching,and sentence repetition tests,are effective methods for assessing the speech characteristics of MCI patients.These tasks involve asking patients to repeat specific content,and analyzing the accuracy of their repetitions to assess their speech function.The application of machine learning and deep learning techniques enables automatic extraction of MCI-related features from speech data,improving diagnostic accuracy.The combined application of these techniques contributes to the early detection of MCI,providing a basis for timely intervention.
4.Transient neonatal myasthenia gravis: a case report and literature review
Li HOU ; Linlin YIN ; Lin JIANG ; Jing ZHAO
Chinese Journal of Neonatology 2024;39(4):219-223
Objective:To summarize the clinical features of transient neonatal myasthenia gravis (TNMG).Methods:We retrospectively collected the clinical data of an neonate diagnosed with TNMG in the department of Neonatology at the Affiliated Hospital of North Sichuan Medical College. Literature was retrieved from databases including CNKI, VIP, Wanfang, the Chinese Medical Journal Full Text database, the Chinese biomedical literature database, PubMed, Embase, Web of Science, The Cochrane library, and Medline. Search terms were "neonatal"、"pregnancy"OR"pregnant"OR"maternal"、"myasthenia"OR "myasthenia gravis"、"transient myasthenia gravis"OR"transitory myasthenia gravis"OR"temporary myasthenia gravis"、"congenital myasthenia gravis"OR"geneogenous myasthenia gravis". Searching period was from establishment of database to May 31st 2023. We summarize the clinical characteristics, treatment, and prognosis of reported cases with TNMG.Results:This patient is a full-term male infant who experienced decreased muscle tone, weak crying, and difficulty breathing 10 min after birth. His mother had a history of myasthenia gravis for 13 years. On the second day of birth, his neostigmine test was positive. So he was diagnosed with TNMG and treated with respiratory support and intravenous human immunoglobulin. His symptoms were relieved at day 10 and resolved by day 20. No abnormalities were observed during follow-up up to 6 months of age. A total of 7 Chinese papers and 13 English papers were retrieved, reporting 39 cases in total including this patient. Mothers of 34 cases (87.2%) were diagnosed with MG before delivery, and the other 5 mothers (12.8%) were diagnosed with MG after delivery. The main clinical manifestations in neonates were reduced muscle tone (29 cases, 74.4%), weak crying (23 cases, 59.0%), dyspnea (20 cases, 51.3%), and feeding difficulties (18 cases, 46.2%). 33 cases (84.6%) developed symptoms within the first day after birth, 6 cases (15.4%) had symptoms between day 2 and day 4. Most patients had a disease course for 2-6 weeks (29 cases, 74.4%). 27 cases (69.2%) underwent neostigmine test, all were positive. 30 cases (76.9%) were treated with cholinesterase inhibitors, 5 cases (12.8%) received IVIG and 1 case (2.6%) received an exchange transfusion. Out of the 39 cases, 33 cases (84.6%) were cured and discharged, 6 cases (15.4%) died. The causes of death were pulmonary infection and respiratory failure. Among the 17 patients with follow-up records, except for 1 case who only followed up to 2 months old and had slightly worse head upright, the rest of the children had normal physical and neurological development.Conclusions:TNMG may occur in infants that were born to MG mothers, most of the patients with TNMG develop symptoms on the first day after birth, a few patients develop symptoms 3-4 d after birth. Neostigmine test, serum MG antibody test should be performed to assist diagnosis. Cholinesterase inhibitors and symptomatic support are the main treatment options. TNMG generally have a good prognosis after treatment.
5.Advances in radiotherapy timing for brain metastases
Jianhua FENG ; Linlin ZHENG ; Yimin LI ; Jun YIN
Chinese Journal of Radiation Oncology 2024;33(4):360-364
Brain metastasis is the most common intracranial tumor in adults. Radiotherapy is one of the most important treatments, in addition to drugs, surgery, and other treatments. In recent years, rapid development of targeted therapy and radiosurgical technology has transformed the treatment mode of brain metastases, with increasing emphasis on individualized treatment. However, the mode of radiotherapy in patients with driver gene-positive remains controversial. In this article, the advances in the timing of radiotherapy for brain metastases were reviewed.
6.Association between dietary habits and sleep duration among middle school students in Shanghai
YIN Xiaoya, ZHAI Yani, YUAN Linlin, YAN Qiong, ZHOU Xinyi, LUO Chunyan, SUN Lijing
Chinese Journal of School Health 2024;45(8):1140-1143
Objective:
To explore the association between dietary behaviors and sleep duration among middle school students in Shanghai, so as to provide reference for interventions targeting insufficient sleep.
Methods:
From May to June 2021, a stratified cluster random sampling method was employed to select a sample of 10-17yearold middle school students for monitoring their healthrisk behaviors. A total of 5 538 valid questionnaires were collected. The survey included items such as daily sleep duration, weekly consumption of sugary beverages, freshly squeezed fruit juice, fresh fruits, fresh vegetables, fried foods, milk and yogurt, breakfast habits, and frequency of eating outside. Statistical analysis was conducted using Chisquare test, Wilcoxon ranksum test, and multivariable Logistic regression model.
Results:
About 73.7% of middle school students reported insufficient sleep in Shanghai. There was a positive correlation between the average daily consumption of fresh fruits and breakfast consumption with sleep duration. In other words, a higher frequency of consuming fresh fruits (OR=1.29) and eating breakfast (OR=1.07) were associated with a higher likelihood of sufficient sleep. Conversely, there was a negative correlation between the frequency of consuming desserts (OR=0.78) and fried foods (OR=0.88) and sleep duration (P<0.05).
Conclusions
Increasing the consumption of fresh fruits and maintaining regular breakfast habits while reducing the intake of fried foods can contribute to achieving sufficient sleep among middle school students. When implementing interventions to improve sleep among middle school students, promoting healthy and balanced diets can be considered as one of the intervention strategies.
7.Effect of LncRNA OIP5-AS1 on Phenotypic Transformation of IOSE80 in Ovarian Epithelium
Linlin SONG ; Huanran MENG ; Lina ZHOU ; Rui LIU ; Lijun YIN
Chinese Journal of Modern Applied Pharmacy 2024;41(5):649-656
OBJECTIVE
To explore the phenotypic changes and possible mechanisms of lncRNA OIP5-AS1 on the proliferation, migration, apoptosis, invasion and cycle of ovarian epithelial cells IOSE80.
METHODS
The clinical data of patients were collected from TCGA database and GEO database. After R package analysis, the differential expression of OIP5-AS1 was visualized in the volcanic map. The correlation between survival rate and OIP5-AS1 was analyzed by Kaplan-Meier. The IOSE80 cell model of OIP5-AS1 over expression and silencing was constructed with lentivirus vector. The expression of OIP5-AS1 was verified by RT-qPCR. Cell proliferation was detected by CCK-8. Invasion was detected by Transwell. Cell migration was detected by scratch test. Cell cycle and apoptosis were detected by flow cytometry. Western blotting was used to detect the expression of E-cadherin and N-cadherin, as well as the expression of cyclin-dependent kinase(CDK) and cyclin-G-related kinase(GAK).
RESULTS
RT-qPCR results showed that IOSE80 cell lines over expressing and silencing OIP5-AS1 were successfully constructed. CCK-8 results showed that overexpressing OIP5-AS1 promoted the proliferation of IOSE80 cells. Scratch test results showed that overexpressing OIP5-AS1 promoted the migration of IOSE80 cells. Transwell results showed that overexpressing OIP5-AS1 would increase the invasiveness of IOSE80 cells. Flow cytometry results showed that overexpression of OIP5-AS1 weakened the apoptosis of IOSE80 cells and promoted the progress of cell cycle. Western blotting results showed that overexpression of OIP5-AS1 downregulated the expression of E-cadherin and upregulated the expression of N-cadherin, while overexpression of OIP5-AS1 increased the expression of CDK and GAK proteins.
CONCLUSION
LncRNA OIP5-AS1 further interferes with the regulation of IOSE80 cell cycle by up regulating the expression of CDK and GAK, and then indirectly regulates the malignant phenotype of ovarian epithelial cells.
8.Epidemiological characteristics of human respiratory syncytial virus in Hubei Province from 2016 to 2023
Qiangling YIN ; Xiao YU ; Xiang LI ; Bin FANG ; Xiaolu ZHANG ; Mingwei PENG ; Guojun YE ; Linlin LIU
Chinese Journal of Preventive Medicine 2024;58(8):1129-1134
Objective:To analyze the epidemiological characteristics of human respiratory syncytial virus (HRSV) in patients with acute respiratory infection (ARIs) in sentinel hospitals of the Hubei influenza surveillance network from 2016 to 2023.Methods:ARIs samples [including influenza-like cases (ILI) and severe acute respiratory infection (SARI)] were collected from influenza surveillance sentinel hospitals in Hubei Province from 2016 to 2023, and case information was collected. HRSV virus nucleic acid typing was performed by fluorescence quantitative PCR method, and the data were collated, plotted and analyzed.Results:From 2016 to 2023, 12 779 cases of ILI and 9 166 cases of SARI were collected. The positive rate of HRSV was the highest in<5 years of age group [15.77% (168/1 065)], among which the positive rate was the highest in 2 to 5 years of age group of ILI cases [13.60% (31/228)], and the positive rate was the highest in 0 to 2 years of age group of SARI cases [25.97% (60/231)] (all P values<0.001). The positive rate of HRSV in SARI cases was 2.31%-25.97%, higher than that in ILI cases (0-13.60%) ( P=0.016). HRSV was prevalent in autumn and winter from 2016 to 2020 and in spring in 2023. Alternating epidemics of HRSV virus type A and B in Hubei Province from 2016 to 2023 (dominant epidemics of type B in 2016 and 2020; dominant epidemics of type A in 2017-2019 and 2023). Conclusion:SARI and ILI patients under five years old are the main infection groups of HRSV. The seasonal prevalence characteristics of HRSV in Hubei Province from 2016 to 2023 shift from autumn and winter to spring.
9.Epidemiological characteristics of human respiratory syncytial virus in Hubei Province from 2016 to 2023
Qiangling YIN ; Xiao YU ; Xiang LI ; Bin FANG ; Xiaolu ZHANG ; Mingwei PENG ; Guojun YE ; Linlin LIU
Chinese Journal of Preventive Medicine 2024;58(8):1129-1134
Objective:To analyze the epidemiological characteristics of human respiratory syncytial virus (HRSV) in patients with acute respiratory infection (ARIs) in sentinel hospitals of the Hubei influenza surveillance network from 2016 to 2023.Methods:ARIs samples [including influenza-like cases (ILI) and severe acute respiratory infection (SARI)] were collected from influenza surveillance sentinel hospitals in Hubei Province from 2016 to 2023, and case information was collected. HRSV virus nucleic acid typing was performed by fluorescence quantitative PCR method, and the data were collated, plotted and analyzed.Results:From 2016 to 2023, 12 779 cases of ILI and 9 166 cases of SARI were collected. The positive rate of HRSV was the highest in<5 years of age group [15.77% (168/1 065)], among which the positive rate was the highest in 2 to 5 years of age group of ILI cases [13.60% (31/228)], and the positive rate was the highest in 0 to 2 years of age group of SARI cases [25.97% (60/231)] (all P values<0.001). The positive rate of HRSV in SARI cases was 2.31%-25.97%, higher than that in ILI cases (0-13.60%) ( P=0.016). HRSV was prevalent in autumn and winter from 2016 to 2020 and in spring in 2023. Alternating epidemics of HRSV virus type A and B in Hubei Province from 2016 to 2023 (dominant epidemics of type B in 2016 and 2020; dominant epidemics of type A in 2017-2019 and 2023). Conclusion:SARI and ILI patients under five years old are the main infection groups of HRSV. The seasonal prevalence characteristics of HRSV in Hubei Province from 2016 to 2023 shift from autumn and winter to spring.
10.Correlation between stress and Internet addiction among middle school students in Shanghai
YUAN Linlin, ZHANG Zhe, ZHOU Xinyi, ZHAI Yani, YIN Xiaoya, LI Shuman, SUN Lijing
Chinese Journal of School Health 2024;45(12):1757-1760
Objective:
To understand the relationship between stress and Internet addiction among middle school students in Shanghai, so as to provide a scientific basis for promoting students mental health and preventing Internet addiction.
Methods:
From May to June 2021, a stratified random cluster sampling method was used to select 6 123 middle and high school students in Shanghai for health risk behavior monitoring. Daily Stressors Evaluation Scale for Urban Secondary School Students was used to evaluate students' stress, and the Internet Addiction Test compiled by Young was used to evaluate students Internet addiction. The correlation between student stress and Internet addiction was analyzed by Kruskal-Wallis H test , Chi square test and multiple Logistic regression.
Results:
Total stress score of middle school students in Shanghai was 24 (12, 39), academic stress score was 8 (5, 13), physical and psychological stress score was 6 (2, 10), interpersonal stress score was 5 (1, 9), and family stress score was 4 (1, 8). The detection rate of Internet addiction was 4.7%. Multivariate Logistic regression analysis showed that the risk of Internet addiction among middle school students with high levels of stress was 8.05 times(95% CI =4.59-14.12) that of students with low levels of stress( P <0.05). The risk of Internet addiction among middle school students with high levels of academic stress, physical and psychological stress, interpersonal stress and family stress was 5.98(95% CI =3.69-9.70), 6.92(95% CI =4.03-11.88), 4.85(95% CI =3.11-7.55), and 4.18(95% CI =2.73-6.40) times that of students with low levels of stress, respectively( P <0.05).
Conclusion
The academic stress, physical and psychological stress, interpersonal stress, and family stress among middle school students can all lead to an increased risk of Internet addiction.


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