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Intelligent control systems can effectively assist in the construction and management of laboratory animal facilities, improving operational efficiency, ensuring the reliability of animal experimental results, and significantly saving human resources. The intelligent control system for laboratory animal facilities at Shenzhen Institute for Drug Control was completed in April 2021. It includes an intelligent management platform and an information management system for animal laboratories. The intelligent management platform regulates room environment parameters such as temperature, humidity, and pressure through building equipment management system, controlling devices such as the Venturi valve, electric air valve, electric water valve, and steam humidification valve. At the same time, various environmental parameters are monitored online through the environmental monitoring system. The laboratory’s intelligence is further enhanced by systems such as automatic lighting control, full HD video monitoring, automatic access control and door system, independent ventilation and feeding, automatic cleaning, automatic exhaust gas treatment, centralized gas supply, and real-time instrument parameter monitoring. The information management system for animal laboratories integrates inspection, instrument and equipment, personnel, documents, standard substances, reagents, inspection standards, books, records, scientific research management, relevant applications, quality management, and query statistics. For animal experimentation, a management module has been developed to achieve a comprehensive digitization of animal management. Furthermore, real-time collection and recording of data such as balance calibration, sample quality, and animal weight are facilitated through electronic experimental recording. In summary, the Animal Laboratory of Shenzhen Institute for Drug Control has extensively utilized intelligent systems to achieve real-time online control and monitoring, improve efficiency, ensure high-quality facility operation, and meet standard requirements. Smooth execution of all inspection and research activities has been achieved over the past three years. This paper provides insights into the construction, management, and operation of laboratory animal facilities at Shenzhen Institute for Drug Control, offering guidance for the implementation of intelligent control in similar facilities across China.
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Objective:To investigate the tau deposition pattern in Alzheimer′s disease (AD) and its correlation with cognition by 18F-MK6240 PET imaging. Methods:From August 2021 to February 2022, 46 elderly people over 55 years old (16 males, 30 females; age (68.9±7.7) years) were included from outpatient and community in Shanghai. Structural brain MRI, β-amyloid (Aβ) PET imaging, tau-PET imaging and comprehensive neuropsychological tests batteries were conducted. The subjects were divided into AD group ( n=16) and normal cognition (NC) group ( n=30) according to the 2018 National Institute on aging and Alzheimer′s Association (NIA-AA) diagnostic criteria. Quantitative analysis was conducted to investigate the tau deposition pattern in AD after preprocessing 18F-MK6240 PET images with MRI images. SUV ratio (SUVR) of brain regions such as entorhinal cortex, hippocampus, parahippocampal gyrus, amygdala, insular lobe, frontal lobe, precuneus, occipital lobe, thalamus and putamen were analyzed, with cerebellum as reference region. The differences of tau deposition in brain regions between AD and NC groups were analyzed by independent-sample t test. The associations between SUVR and Mini-Mental State Examination (MMSE) score and Montreal Cognitive Assessment-Basic (MoCA-B) score were analyzed by Pearson correlation analysis. Results:AD displayed a significant tau deposition in frontal lobes, temporal lobes and parietal lobes compared with NC. SUVR of brain regions in AD group were higher than those in NC group ( t values: 3.37-9.61, all P<0.05). SUVR in brain regions were negatively correlated with MMSE score ( r values: from -0.735 to -0.350, all P<0.05) and MoCA-B score ( r values: from -0.723 to -0.367, all P<0.05). Conclusion:18F-MK6240 PET can demonstrate the tau deposition in the brain of AD patients, and the tau deposition is related to cognitive function.
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Objective:To explore the β-amyloid (Aβ) deposition pattern of subjects with the preclinical Alzheimer′s disease (AD), community-derived amnestic mild cognitive impairment (aMCI) and normal cognition (NC) from communities of Shanghai.Methods:According to the inclusion and exclusion criteria, 273 subjects (104 males, 169 females; age (64.2±7.6) years) were recruited from Shanghai community and memory clinics from December 2018 to July 2020. All subjects underwent MRI, 18F-AV45 PET imaging and neuropsychological scale tests and were grouped into AD, aMCI and NC groups based on clinical diagnosis. Differences in demographic information, the neuropsychological scale tests′ scores and positive rate of Aβ deposition among each group were analyzed by one-way analysis of variance or χ2 test. Aβ deposition patterns of AD and MCI groups were analyzed at voxel level, and the differences of Aβ deposition among different groups were compared. Results:Among 273 patients, the positive rates of Aβ deposition in AD, aMCI and NC groups were 84.4%(38/45), 36.4%(20/55) and 23.1%(40/173), respectively ( χ2=58.37, P<0.001). Among AD, aMCI, NC and NC (Aβ-) groups ( n=132), the education years of AD group was the lowest ((9.7±4.6) years; F=8.86, P<0.001). In addition, there were significant differences in the scores of several neuropsychological scale tests among AD, aMCI, NC groups and NC (Aβ-) group ( F values: 27.68-235.50, all P<0.001). Compared with subjects in NC(Aβ-) group, the Aβ depositions in the aMCI and AD groups were widely distributed in the whole cerebral cortex; and AD group had higher Aβ deposition in bilateral frontal, parietal, temporal, occipital lobe, cingulate gyrus and precuneus than aMCI group. Conclusions:The positive rate of Aβ deposition in the preclinical AD population from the Shanghai community is obtained. There are significant different Aβ deposition patterns in subjects at different stages of AD.
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Objective:To explore the association of the impaired cognition and the deposition of β-amyloid (Aβ) in normal cognitive (NC) and mild cognitive impairment (MCI).Methods:From December 2018 to January 2021, 305 subjects (113 males, 192 females; age (64.0±7.7) years) who completed neuropsychological tests and MRI in Shanghai Sixth People′s Hospital, Shanghai Jiao Tong University and 18F-florbetapir (AV45) PET imaging in Huashan Hospital, Fudan University were retrospectively analyzed. The subjects were divided into MCI group and NC group based on neuropsychological tests, and each group was further divided into Aβ-positive and Aβ-negative based on PET imaging results. Independent-sample t test, Mann-Whitney U test and χ2 test were used to analyze the data. Results:There were 118 subjects in MCI group and 187 subjects in NC group. The Aβ-positive rate in MCI group (37.3%, 44/118) was higher than that in NC group (26.2%, 49/187; χ2=4.19, P=0.041). The assessment performances of MCI group in general cognitive function, memory function, language function and executive function were inferior to those of NC group ( t values: from -10.63 to -6.31, z values: from -11.01 to -6.03, all P<0.001). The Auditory Verbal Learning Test-Long Delay Recall (AVLT-LDR) score of Aβ-positive subjects was lower than that of Aβ-negative subjects in MCI group (1.00(0.00, 3.00) and 3.00(1.00, 4.00); z=-2.49, P=0.013). The Montreal Cognitive Assessment Basic (MoCA-B) score of Aβ-positive subjects was lower than that of Aβ-negative subjects in NC group (25.29±2.67 and 26.36±2.42; t=-2.61, P=0.010). Conclusion:Compared to Aβ-negative subjects, MCI patients with Aβ-positive perform worse on memory tests, and NC subjects with Aβ-positive perform worse on general cognitive function.
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Objective:To explore the abnormal brain metabolic pattern and connectivity in temporal lobe epilepsy (TLE) patients.Methods:18F-FDG PET images of 75 patients diagnosed as drug resistant unilateral TLE from January 2014 to December 2016 in Huashan Hospital of Fudan University were collected retrospectively, including 41 (22 males, 19 females, age (28.4±8.7) years) left TLE (LTLE) and 34 (13 males, 21 females, age (28.5±8.8) years) right TLE (RTLE). Forty-four healthy controls (24 males, 20 females, age (31.2±6.2) years) were also enrolled. The cerebral glucose metabolism in TLE patients and the controls were analyzed with statistical parametric mapping (SPM) 12. The brain connectivity based on glucose metabolism were analyzed with bilateral hippocampus and amygdala as seeds. Permutation test with 1 000 permutations was used to analyze data. Results:Compared to control group, in both LTLE and RTLE groups, hypometabolism was found in affected hippocampus, amygdala, insula and temporal gyrus and hypermetabolism was observed in health hippocampus, parahippocampal gyrus, amygdala, lenticular nucleus and thalamus. In addition, hypometabolism was also found in affected superior/middle frontal gyrus and hypermetabolism was also found in bilateral frontal-orbital gyrus, bilateral cerebellum, affected lenticular nucleus and thalamus in LTLE group. In both TLE groups, affected seeds exhibited increased connectivity with affected superior frontal gyrus, lingual gyrus, fusiform gyrus, superior/middle temporal gyrus and temporal pole (all P<0.05); affected seeds exhibited increased connectivity with health superior frontal gyrus ( P=0.005), lingual gyrus ( P=0.018) and transverse temporal gyrus ( P=0.016) in RTLE group in addition. Besides, affected seeds exhibited decreased connectivity with bilateral default mode network (DMN) (all P<0.05), affected caudate nucleus ( P=0.015) and health thalamus ( P=0.008), in a uniform distribution pattern in LTLE group, and with bilateral cerebral cortex in an irregular distribution pattern in RTLE group (all P<0.05). In LTLE group, health seeds exhibited more increased connections with superior ( P=0.005)/middle frontal gyrus ( P=0.042), health hippocampus ( P=0.038), parahippocampal gyrus ( P=0.019), amygdala ( P=0.038), posterior cingulate gyrus ( P=0.004), and bilateral fusiform gyrusand ( P=0.048) compared with RTLE group; while, in RTLE group, health seeds exhibited more decreased connections with health superior ( P=0.047), inferior frontal gyrus ( P<0.001), orbital frontal gyrus ( P<0.001) and rectus gyrus ( P=0.016) compared with LTLE group. Conclusion:Altered brain glucose metabolism and connectivity pattern are found and will elucidate the underlying metabolic pattern of TLE.
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Objective:To investigate the expression of miR-527 in bladder cancer (BC) and its effect on the proliferation, migration and invasion of bladder cancer cells.Methods:From February 2018 to June 2019, the immortalized human bladder epithelial cell line SV-HUC-1 and human bladder cancer cell lines T24, UM-UC-3, 5637 and RT-112 were cultured in vitro. Real time quantitative PCR (qRT-PCR) was used to detect the expression of miR-527 in BC bladder cancer tissues and adjacent normal bladder tissues, human bladder cancer cell lines and human bladder epithelial immortalized cell lines. MiR-527 mimics, miR-527 inhibitor, ENO1 overexpression plasmid, ENO1 siRNA and corresponding negative control were transfected into bladder cancer cell line. CCK8 test, clone formation test and Transwell test were used to study the cell proliferation, migration and invasion. Luciferase reporter gene assay was used to verify the target gene of miR-527. Western blotting was used to analyze the regulation of miR-527 on target gene expression.Result:Compared with normal bladder tissue, the expression of miR-527 in bladder cancer was significantly lower (1.723±1.070 vs. 1.148±0.760, P<0.05). The relative expression of miR-527 in T24 (0.540±0.082), UM-UC-3 (0.230±0.053), 5637(0.463±0.085) and RT-112 (0.310±0.056) were significantly lower than those in SV-HUC-1 cells (0.987±0.111) with statistical significance ( P<0.05). Compared with the negative control (NC) group, CCK8 assay results showed that the cell viability was significantly decreased after transfection of miR-527 mimics into UM-UC-3 cells ( P<0.05). The clone formation test showed that the number of cell clones in UM-UC-3 cells transfected with miR-527 mimics was significantly lower than that in the control group (157.00±15.52 vs 57.33±15.50, P<0.05). Compared with the control group, the cell activity of T24 cells transfected with miR-527 inhibitor was significantly increased ( P<0.05). Compared with the control group, the number of cell clone formation was significantly increased (76.67±9.07 vs. 141.70±10.50, P<0.05). According to the prediction of targetscan database, ENO1 was the target gene of mir-527. Luciferase reporter gene experiment showed that the luciferase activity of mir-527 mimics group was significantly lower than that of control group (0.99±0.02 vs. 0.47±0.10, P<0.05), while the luciferase activity of miR-527 mimics group was significantly lower than that of control group (0.99 ± 0.02 vs. 0.47 ± 0.10, P<0.05), without statistical significance (1.03±0.04 vs. 0.96±0.05, P>0.05). Western blot analysis showed that the expression of ENO1 in miR-527 mimics group was significantly lower than that in NC mimics group (1.09±0.17 vs. 0.31±0.13, P<0.05), and the expression of ENO1 in miR-527 inhibitor group and NC inhibitor group were significantly increased (0.97±0.09 vs. 2.17±0.15, P<0.05). Compared with miR-527 mimics group, transfection of miR-527 mimics+ ENO1 overexpression plasmid could reduce the inhibitory effect of miR-527 mimics on proliferation, migration and invasion of bladder cancer cell line ( P<0.05). Compared with miR-527 inhibitor group, transfection of miR-527 inhibitor+ ENO1 siRNA could weaken the inhibit effect of miR-527 on the proliferation, migration and invasion of bladder cancer cell lines ( P<0.05). Conclusion:miR-527 is low expressed in BC and can be used as a tumor suppressor gene to inhibit the proliferation, migration and invasion of BC cells.
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Objective To optimize the detection test of Pig-a gene mutation in peripheral blood of rats by enriching and detecting mutant erythrocytes, using immunomagnetic separation technique in combination with flow cytometry. Methods SD rats were administered with 20,40 and 80 mg/(kg·bw)doses of N-ethyl-N-nitrosourea(ENU) continually for 3 days. The peripheral blood samples of rats were collected on the 7th,14th and 28th days, respectively, after treatment. Immunomagnetic separation columns were used to enrich RETCD59- and RBCCD59- cells, and then flow cytometry was used to count the number of pre-column and post-column peripheral erythrocytes. Results Compared with the control group,the frequencies of RETCD59- and RBCCD59- were significantly increased in each ENU group(P<0.05). With immunomagnetic separation technique, the test of Pig-a gene mutation of a sample could be completed within 3 minutes,and the number of detected RETCD59- or RBCCD59-cells was up to 2×104or 9×104, respectively. Conclusions In this study,immunomagnetic separation in combination with flow cytometry is used to establish and optimize the Pig-a gene mutation test in rat peripheral blood,showing a high-throughput detection and improved accuracy and efficiency of the experiment.
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Skin sensitization (allergic contact dermatitis, ACD), is a serious condition caused by small reactive molecules and is characterized by a delayed-type hypersensitivity .Animal tests were usually used in the evaluation of sensitizing potential of chemical substances in the past .However , there is an increasing interest from the public for reducing and ultimately replacing animal tests .The European Union (EU) has posed a ban on animal testing of cosmetic ingredients that includes skin sensitization since 2013.Therefore, alternative in vitro tests are the main tendency in chemical substances and cosmetic sensitizing potential research in the future .In this study, different kinds of in vitro test methods that were adopted by OECD or on research (LLNA, DPRA, KeratinoSens TM, h-CLAT) were reviewed through recent years literature , comprehensive introduction and evaluation were made to obtain reliable hazard and potency information on potential skin sensitizers .
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Objective This review was aimed to provide reference for production, management and use of laboratory animals by analyzing the test results on intestinal parasitic infections of mice and rats in different provinces from 1989 to 2013 in China.The results showed that the infection rates in clean and SPF mice and rats were reduced to 10%, being better than that in the past years, but the situation was still not optimistic for the control of flagellate parasites infections.
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To promote the understanding and practice of evidence-based medicine (EBM) in coronary artery bypass grafting (CABG). The transition of CABG graft materials from saphenous vein to arterial conduits were perceived using the principles of EBM. The arterial conduits were increasingly performed based on the principles and practice of EBM, therefore the postoperatively long-term graft patency rate, survival rate, and free cardiac event rate were significantly improved. CABG outcomes can be significantly improved by using arterial conduits based on principles of EBM.
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Objective To investigate the relationship of the activation of nuclear factor kappa B(NF-kB) with myocardial neutrophil infiltration and injury in human open heart surgery,and to observe the inhibiting effect on the activation of NF-kB and protecting effect on myocardium of the coenzyme Q10,a scavenger of oxygen free radicals.Methods Forty-seven adult patients undergoing open heart surgery were randomly divided into two groups,the control and the treatment.CoenzymeQ10 tablets were given to the treatment group 5 days before operation.Biopsy of right atrium for myocardial pathology,activated NF-kB detection and ultrastructure observation were done prior to cardiopulmonary bypass,45 minutes of ischemia and 45 minutes of reperfusion.The dynamic indexes,vasomotor drug dosage,and outcomes were observed postoperatively.Results Upon 45 minutes of ischemia and 45 minutes of reperfusion,in control group there were neutrophil accumulation and adhesion of vascular endothelium,ultrastructural damages,and positive expression of NF-kB both in nuclei and cytoplasm,in myocardium.In treatment group,there were only mild neutrophil infiltration and ultrastructural damages,and weak positive expression of NF-kB both in nuclei and cytoplasm.However,the dynamic indexes,vasomotor drug dosage,and outcomes of two groups were not significantly different.Conclusion NF-kB plays an important role in pathophysiological process of myocardial ischemia and reperfusion in open heart surgery.CoenzymeQ10 has obvious inhibiting effect on activation of NF-kB and protecting effect on myocardium.