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.

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.

Objective To investigate the effects of fructose exposure on mitochondrial oxidative damage in M2-type macrophages and elucidate the regulatory role of protein phosphatase 2A(PP2A)in the process using its specific activator ABL127,an inhibitor of protein phosphatase methylesterase-1(PPME-1).Methods ① Immortalized mouse bone marrow-derived macrophages Ana-1 were subjected and grouped into M0(conventional culture),M2(treated with 20 ng/mL IL-4 for 24 h),and M2+Fru groups(IL-4 plus 0.04,0.20,1.00,or 5.00 mmol/L fructose).Cell viability was assessed with CCK-8 assay.Number of mitochondria,total and mitochondrial levels of reactive oxygen species(ROS),and mitochondrial membrane potential(ΔΨM)were measured using fluorescent probes.Total and demethylated PP2Ac protein levels were detected by Western blotting.② Ana-1 cells were also divided into M0,M2,M2+Fru(20 ng/mL IL-4+5.00 mmol/L fructose,24 h),and M2+Fru+ABL127(20 ng/mL IL-4+5.00 mmol/L fructose+1.00 μmol/L ABL127,24 h)to investigate PP2A-mediated mechanisms.Numbers of mitochondria and lysosomes,ROS level,and ΔΨM were detected via fluorescence assays.Expression of mitophagy-related proteins,PTEN induced putative kinase 1(PINK1),P62,microtubule-associated protein light chain 3(LC3),and voltage-dependent anion channel(VDAC)was evaluated by Western blotting,and the mRNA levels of M2 markers,found in inflammatory zone 1(Fizz1),arginase-1(Arg-1),and TGF-β were measured using RT-qPCR.Results ① Compared with the M2 group,fructose treatment at a concentration ranging from 0.04 to 5.00 mmol/L showed no effect on cell viability in M2 macrophages,but increased total ROS level in a dose-dependent manner(P<0.05).Fructose of 5.00 mmol/L resulted in significantly elevated mitochondrial ROS and mitochondrial quantity(P<0.05),reduced ΔΨM(P<0.05),up-regulated demethylated-PP2Ac(P<0.05),and no changed total-PP2Ac protein level.② Compared with the M2+Fru group,the addition of ABL127 led to decreased number of mitochondria but increased number of lysosomes(P<0.01),up-regulation of PINK1,LC3Ⅱ and VDAC proteins,down-regulation of P62(P<0.05),reduced total and mitochondrial ROS levels,and enhanced ΔΨM(P<0.01).The mRNA expression of Fizz1,Arg-1,and TGF-β was notably decreased in the M2+Fru group than the M2 group(P<0.05),and the levels were rescued by ABL127 treatment(P<0.05).Conclusion Fructose induces PP2Ac demethylation and then mitochondrial oxidative damage in M2-type macrophages.PP2A activation promotes mitophagy and reverses fructose-induced damage.

Objective:To construct an intervention program of sports and medical integration for patients with acute myocardial infarction (AMI) in phase Ⅱ cardiac rehabilitation, so as to provide reference for the effective implementation of physical activity behavior change in AMI patients during phase Ⅱ cardiac rehabilitation.Methods:Before the research, we browsed relevant literature and guidelines published from July 2016 to June 2021, made a retrospective study on the influencing factors of cardiac rehabilitation behavior for AMI patients, and carried out a qualitative interview on cognitive and compliance motivations for AMI patients. Furthermore based on trans-theoretical model, the first draft of the intervention program was developed and and the expert consultation questionnaire was formed. From February to April 2022, Delphi method was used in 16 experts from 10 hospitals and 1 nursing college in 4 provinces. After 2 rounds of expert consultations on the importance and operability of items, the intervention program was finally determined.Results:The effective recovery rates of the 2 rounds of expert consultations questionnaires were both 16/16. The authority coefficient of expert consultation was 0.90, the judgment basis coefficient was 0.96, and the familiarity degree was 0.84. After the second round of expert consultation, the coefficient of variation of the importance of each item was (0.0-13.4)%, and the coefficient of variation of operability was (0.0-18.1)%. The final intervention program of sports and medical integration for patients with AMI in phase Ⅱ cardiac rehabilitation had 37 items, containing precontemplation stage (8 items), contemplation stage (7 items), preparation stage (5 items), action stage (9 items) and maintenance stage (8 items).Conclusions:The construction process of the intervention program of sports and medical integration for patients with AMI in phase Ⅱ cardiac rehabilitation is scientific and feasible. The content is focused on the patient-centred conception and the whole-process management for the exercise rehabilitation of AMI patients in phase Ⅱ cardiac rehabilitation. This intervention program may improve the safety, feasibility, participation and compliance in phase Ⅱ cardiac rehabilitation in patients with AMI. So it is recommended to be popularized and used in phase Ⅱ cardiac rehabilitation.

Objective To investigate the molecular mechanism of taurine regulating the polarization of M2 macrophages by mitophagy. Methods THP-1 cells were divided into four groups: M0 group (THP-1 cells were treated by 100 nmol/L phorbol myristate ester for 48 hours to polarize into M0), M2 group (THP-1 cells were induced to polarize into M2 macrophages by 20 ng/mL interferon-4 (IL-4) for 48 hours), M2 combined with taurine groups (added with 40 or 80 mmol/L taurine on the basis of M2 macrophages). The mRNA expression of mannose receptor C type 1(MRC-1), C-C motif chemokine ligand 22(CCL22) and dendritic cell-specific ICAM-3 grabbing non-integrin (CD209) in M2 macrophages were detected by quantitative real-time PCR. Mitochondrial and lysosome probes were used to detect the number of mitochondria and lysosomes by multifunction microplate reader and confocal laser scanning microscope. The level of mitochondrial membrane potential (MMP) was detected by JC-1 MMP assay kit. The expression of mitophagy-related proteins PTEN-induced putative kinase 1 (PINK1) and microtubule-associated protein 1 light chain 3 (LC3) were detected by Western blot analysis. Results Compared with M0 group, the expression of MRC-1, CCL22, CD209 and PINK1, the number of mitochondria and the level of MMP in M2 group were significantly increased, whereas the number of lysosomes and LC3II/LC3I ratio were decreased. Compared with M2 group, the expressions of MRC-1, CCL22 and CD209, the number of mitochondria and the level of MMP in M2 combined with taurine group dropped significantly while the number of lysosomes was found increased, and the protein expression of PINK1 and LC3II/LC3I ratio were also increased. Conclusions The polarization of M2 macrophages is regulated by taurine to prevent excessive polarization via reducing the level of MMP, improving the level of mitophagy, reducing the number of mitochondria, and inhibiting the mRNA expression of polarization markers in M2 macrophages.
Mitophagy ; Taurine ; Macrophages/metabolism* ; Protein Kinases/metabolism* ; RNA, Messenger

Objective To investigate the protective effect of Baicalin on inflammation induced by lipopolysaccharide in H9C2 cardiomyocytes and its possible mechanism. Methods H9C2 myocardial cells were cultured and pretreated with baicalin at the final concentration of 10, 20, 30 μmol/L for 12 hours, then stimulated with LPS at the final concentration of 1 μg/mL for 6 hours. The control group was treated with the same amount of saline to collect cell samples. CCK-8 (The Cell Counting Kit-8) was used to detect cell activity, enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of interleukin-6 (IL-6), interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α), Western blot was used to detect the protein expression levels of NF-κB p65, p-NF-κB p65, p38 MAPK, p-p38 MAPK, IκBα and p-IκBα. SPSS 23.0 statistical software was used. Independent sample t test was used for comparison between two groups, and one-way ANOVA test was used for comparison among multiple groups. Results The survival rate of myocardial cells in the control group was (93.67 +1.453)％. Compared with the control group, the survival rate of H9C2 myocardial cells induced by LPS decreased (P< 0.05). In the control group, the expression of IL-6 in H9C2 myocardial cells was (49.33 +2.42) pg/mL, the expression of TNF-α was (86.33 +1.85) pg/mL, and the expression of IL-1β was (28.67 +4.66) pg/mL. Compared with the control group, the expression levels of IL-6, IL-1β and TNF-α in H9C2 myocardial cells increased after LPS induction (P< 0.05), while the levels of p-NF-κ B p65, p-p38 MAPK and p-I κ B α protein increased (P< 0.05), while the levels of I κ B α protein decreased (P< 0.05), while the expressions of NF-κ B p65 and p38 MAPK protein did not change significantly (P> 0.05). Compared with LPS group, the survival rate of H9C2 myocardial cells in baicalin intervention group increased (P<0.05), the expression levels of IL-6, IL-1β and TNF-a decreased (P < 0.05), the levels of p-NF-κB p65, p-p38 MAPK, p-I κBα protein decreased (P< 0.05), and the level of IκBα protein increased (P< 0.05), while the expression of NF-κB p65 and p38 MAPK did not change significantly. (P>0.05). Conclusions Baicalin may alleviate LPS-induced cardiomyocyte inflammation by inhibiting the activation of NF-kappa B and p38 MAPK, and improve cell survival.

Objective: To investigate the assessment and treatment strategy of patients with renal cell carcinoma. Methods: The clinical data of 43 patients with renal cell carcinoma and bone metastases admitted to the First Affiliated Hospital of Nanjing Medical University from January 2006 to December 2018 were retrospectively analyzed. The follow-up time was 6 years, with an average age of 55.4 years (21-87 years). There were 29 males, 14 females, 22 cases of limb bone metastasis, 12 cases of spinal metastasis, 9 cases of multiple bone metastasis, 21 cases of Fuhrman grade 1 and 2, 19 cases of T₁, and 20 cases of N₀. All patients were confirmed by postoperative pathological examination or imaging data suggesting that bone metastasis are from renal cell carcinoma. Forty-three patients underwent primary renal surgery, and molecular targeted therapy was used after the operation. The treatment process was smooth, no obvious discomfort, and postoperative pathology showed clear cell carcinoma.22 patients with limb bones metastasis and 12 patients with spinal metastasis included in the study all met the indications for secondary surgery after the disease assessment. After communicating with the patient, 13 patients with limbs metastasis and 6 patients with spinal metastasis received local treatment, including complete resection of the extremities and spinal fixation, the remaining 15 patients and 9 patients with multiple bone metastasis were treated conservatively. There were 19 patients in the local treatment group, 13 patients with limbs bone metastasis, 6 patients with spinal bone metastasis, the average age was 54.9 years, the average diameter of the primary tumor was 4.7 cm. There were 24 patients in the conservative treatment group, 9 patients with limbs metastasis, 6 patients with spinal metastases and 9 cases with multiple bone metastasis, with an average age of 56 years and a primary tumor diameter of 5.6 cm. Limb metastatic lesions were evaluated according to the patient's general condition, bone pain, fracture risk, and bone metastasis. Spinal lesions were evaluated according to Tokuhashi score, Harrington score, Tomita score, vertebral stability assessment, and molecular targeted therapy. Aminokinase inhibitors, conservative treatment with local radiotherapy and bisphosphonate treatment. Results: During the follow-up period, the 1-year overall survival rate of the local treatment group was 100.0%, the 2-year overall survival rate was 89.4%, and the 5-year overall survival rate was 73.7%. The 1-year overall survival rate of the conservative treatment group was 87.5%, and the 2-year overall survival rate was 62.5%. The 5-year overall survival rate was 16.7%. The 2-year and 5-year survival rates of the local treatment group were statistically different (P=0.044, P=0.000) compared with the conservative treatment group. For patients with limb bone metastasis, the 5-year survival rate was significantly higher in patients receiving topical treatment than in the conservative treatment group (P=0.011). For spinal metastasis, spinal pain in the local treatment group was alleviated to varying degrees. No spinal instability and spasticity were observed after follow-up. In the spine patients who received conservative treatment, 3 patients developed paraplegia, which was statistically different from local treatment (P=0.046). Another 9 patients with multiple bone metastases did not undergo local surgery, and all died after multiple organ failure. Conclusions At the same time of molecular targeted therapy, according to the evaluation results, selective treatment of bone metastases with secondary surgical indications, including complete resection of the extremities and spinal fixation, can significantly improve the survival and quality of life of those patients.

Objective The ammonia level in air and heavy metals in drinking water were explored in a cynomolgus monkey feedlot. Methods Air ammonia from different communities and feeding patterns of cynomolgus monkeys were collected at three time-points per day and determined by Nessler's reagent spectrophotometry. Inductively coupled plasma mass spectrometry was applied to detect the level of heavy metals in drinking water from 5 different sampling sites in the feedlot,incluing the outlet of underground water,the water tank,both monkey cages equipped with PVC or iron pipes and sewage lagoon,respectively. Results Air ammonia levels in quarantine inspection flock cages(0.59 ± 0.03 mg/m3)were significantly higher than in the cages of both reproductive flock(0.34 ± 0.03 mg/m3)and sale flock(0.27 ± 0.04 mg/m3). The ammonia level in air in different feeding patterns ranks as following:cage rearing of quarantine inspection flock(0.59 ± 0.03 mg/m3)>cage rearing of reproductive flock(0.48 ± 0.02 mg/m3)>captive bleeding of sale flock(0.30 ± 0.02 mg/m3)>cage rearing of sale flock(0.25 ± 0.01 mg/m3)> captive bleeding of reproductive flock(0.22 ± 0.02 mg/m3). The air ammonia concentrations of both former flocks were statistically higher than the latter three flocks. The highest air ammonia level among different flocks and feeding patterns occurred in the morning, before waste discharge clean-up. The iron concentration in drinking water in the cages equipped with iron pipes was higher than Chinese drinking water standard. Conclusions The air ammonia level was lower than the Chinese air quality standard. The iron concentration in drinking water in the cages equipped with iron pipes was higher than the Chinese drinking water standard.

Colorectal cancer is caused by the interaction of genetic and environment factors. Domestic and foreign scholars have attempted to develop several colorectal cancer risk prediction models, in order to identity risk factors, to screen for high risk population and evaluate the risk of developing colorectal cancer, so as to provide personalized screening protocols for individuals with different risk, and eventually reduce the incidence and mortality rate of colorectal cancer. Currently, the common colorectal cancer risk prediction models were mainly developed based on case-control study and cohort study. Models developed in European and American regions and Asia (excluding China) only include common risk factors, while Chinese models also include hereditary factors on the bases of common risk factors. However, the development and verification of each model are mainly based on local population, whether it can be applied for other population need to be determined. This article reviews the development, validation and evaluation of the risk prediction models, in order to provide a basis for developing more precise risk prediction models for colorectal cancer.
Asian Continental Ancestry Group ; Case-Control Studies ; Cohort Studies ; Colorectal Neoplasms ; Humans ; Incidence ; Risk Assessment ; Risk Factors

Objective: To compare the predictive value of PSS, APACHEII, SAPSII and SOFA in the prognosis evaluation of acute poisoning. Methods: Clinical data (including PSS score, APACHEII score, SAPSII score and SOFA score, within 24 hours after admission) of 231 acute poisoning patients admitted to the emergency intensive care unit EICU of our hospital from January 2015 to October 2016 was retrospectively analyzed. The patients were divided into the survival group and the dead group according to the 28-day clinical outcomes, comparing the differences of clinical data in each group. To analyze the correlation between PSS score, APACHEII score, SAPSII score and SOFA score in each group, comparing the value and the area under the ROC curve of four scoring systems and evaluate the predictive value of the four scoring systems. Results: Comparing with the survival group and the dead group, PSS score, APACHEII score, SAPSII score and SOFA score were significantly different (P<0.01) . PSS score, APACHEII score, SAPSII score and SOFA score were significantly positive correlation (P<0.01) , the area under the ROC curve (AUC) of the four scoring systems were 0.833, 0.887, 0.843 and 0.843 respectively. The area under the ROC curve (AUC) of APACHEII score was higher than PSS score, SAPSII score and SOFA score, the difference was statistically significant (z=2.351, 2.317, 2.217; P=0.019, 0.021, 0.027) , there was no significant difference in the area (AUC) between the three scoring curves (P>0.05) . The cutoff value (cut-off) , sensitivity, specificity and accuracy rates of PSS score, APACHEII score, SAPSII score and SOFA score were (2.5, 93.1%, 50.9%, 61.5%) , (14.5, 82.8%, 75.7%, 77.48%) , (31.5, 77.6%, 76.90%, 77.08%) , (5.5, 77.60%, 74.60%, 75.35%) . Conclusion PSS score, APACHEII score, SAPSII score and SOFA score can evaluate the prognosis of patients with acute poisoning, but the APACHEII score is better than the other three scoring systems in evaluating the prognosis for its evaluation ability and accuracy rate.