1.Construction and gene identification of CSF1R +/-mice
Yuanyuan Zhou ; Chong Liu ; Anqi Wang ; Huiru Zhang ; Jiaqi Qiu ; Mengjuan Zhu ; Jiajie Tu
Acta Universitatis Medicinalis Anhui 2025;60(5):884-889
Objective:
To constructCSF1R+/-mice and to analyze their genotypes, so as to provide animal model basis for disease pathological mechanism and drug target.
Methods :
A linearized targeting vector was designed according to Cre/Loxp system. A Loxp site was inserted upstream of the 5th exon of theCSF1Rgene, and a neomycin resistance box with Loxp sites on both sides was inserted downstream of the 5th exon. The linearized targeting vector was electroporated into embryonic stem cells. The correctly targeted embryonic stem cells were injected into the blastocysts of C57BL/6J mice to obtain chimeric mice, which were bred with Zp3-Cre mice. The newborn mice were numbered 9-14 days after birth and their tails were cut. The DNA of the mice was extracted, and the genotype of the mice was identified by polymerase chain reaction and agarose gel electrophoresis. The expression of CSF1R in mouse macrophages was detected by flow cytometry. The expression of CSF1R in mouse tissues was detected by Western blot.
Results:
The results of agarose gel electrophoresis showed that 453 bp bands were amplified in wild type mice, and 453 bp and 650 bp bands were amplified in heterozygous mice. The results of flow cytometry showed that the expression of CSF1R in peritoneal macrophages and bone marrow-derived macrophages of CSF1R heterozygous mice was lower than that of WT group(P<0.05). The results of Western blot showed that the expression of CSF1R in spleen, kidney and brain tissue of CSF1R heterozygous group was lower than that of WT group(P<0.05).
Conclusion
CSF1R+/-mice are successfully constructed, reproduced and identified, which provides an animal model basis for further revealing the potential mechanism of CSF1R in immune regulation.
2.Construction, breeding, and gene identification of TREM2 knockout mice
Rong Huang ; Xinxin Zhao ; Hui Xue ; Mengjuan Zhu ; Jiajie Tu ; Xinming Wang
Acta Universitatis Medicinalis Anhui 2025;60(6):977-983
Objective :
To construct triggering receptor expressed on myeloid cells 2(TREM2) gene knockout(TREM2-/-) mice using CRISPR/Cas9 technology, to breed TREM2-/- mice and to analyze the genotype of TREM2-/- mice.
Methods :
CRISPR/Cas9 technology was used to selectively knock out exon 2-3 regions of TREM2 gene to construct a TREM2-/- mouse model, and the genetic background of all mice was C57BL/6J. Polymerase chain reaction(PCR) was used to identify the genotype of mice. Quantitative real-time PCR(qPCR) and Western blot were used to detect the expression level of TREM2 in major tissues of mice, and the authenticity and scientific nature of PCR identification results were verified from mRNA level and protein level. According to the sgRNA sequence, the possible off-target sites were predicted on the CCTop website, and the tail DNA of mice was extracted and amplified by PCR and then Sanger sequencing was performed to detect whether there was off-target effect in TREM2-/- mice.
Results :
TREM2-/- mice were successfully constructed by CRISPR/Cas9 technology, and the mice were genotyped. The results of agarose gel electrophoresis showed that the mouse genotype with only 415 bp band amplified was wild type(WT), the mouse genotype of the 449 bp band amplified only was TREM2-/-, and the mouse genotypes amplified with 415 bp and 449 bp double bands were heterozygous. qPCR results showed that compared with WT mice, the mRNA expression of TREM2 in heart and brain tissues of TREM2-/- mice was down-regulated(P-/- mice was reduced(P-/- mice.
Conclusion
TREM2-/- mice are successfully constructed and bred, a reliable genotype identification method is established, the genetic stability of the mouse model is verified, which will provide an important genetic animal model for the study of TREM2 gene function.
3.Pathogenesis of T2DM with cognitive impairment in Baotou area and the improvement effect of GLP-1 receptor agonists
Yan ZHU ; Mengjuan ZHANG ; Yi HE ; Peiqing LIU
Journal of Public Health and Preventive Medicine 2023;34(6):157-160
Objective To analyze the onset characteristics of type 2 diabetes mellitus (T2DM) with cognitive impairment in Baotou area, and study the improvement effect of GLP-1 receptor agonists. Methods A total of 320 patients with T2DM admitted between September 2021 and September 2022 were selected and divided into the observation group with T2DM and cognitive dysfunction and the negative control group without cognitive dysfunction according to their cognitive function status , Among the 160 cases in each group; Patients with type 2 diabetes and cognitive impairment were randomly divided into a treatment group and a control group, 80 cases in each group; the control group was treated with conventional treatment, and the treatment group was additionally treated with semaglutide; Logistics multiple regression model was used to analyze T2DM The related risk factors of cognitive impairment in patients were assessed by the Mini-Mental State Examination (MMSE) score to evaluate the cognitive function of the patients. Results Multivariate regression model showed that course of disease, age, vitamin D, HbA1c, LDL-C, BMI, Hcy, Lp-PLA2, TNF-α, IL-6 and folic acid levels were also independent risk factors for cognitive impairment in T2DM patients (P<0.05); There was a significant positive correlation between GLP-1 receptor agonists and cognitive function recovery in T2DM patients with cognitive impairment (P<0.05). Conclusion The onset of T2DM with cognitive impairment in Baotou area is often accompanied by a long course of disease, older age, abnormal levels of vitamin D, HbA1c, LDL-C, BMI, Hcy, Lp-PLA2, TNF-α, IL-6 and folic acid, and GLP -1 receptor agonists have a clear role in improving the cognitive function of patients.
4.Treatment of major depression disorder and bilateral angle-closure glaucoma comorbidity: a case report
Mengjuan XING ; Zhixin JIN ; Yang SUN ; Liping ZHU
Sichuan Mental Health 2021;34(6):574-576
The purpose of this study is to describe the principles in selection and application of antidepressants in patients with depression complicating glaucoma or at high risk of glaucoma. With the aim of providing a partial reference for relevant issues, this paper elaborated a case of major depression after glaucoma surgery receiving 6 weeks of treatment with escitalopram oxalate and sulpiride achieved significant improvement in depressive and psychotic symptoms without triggering or exacerbating glaucoma.
5.Association of antenatal anxiety with preterm birth and low birth weight: evidence from a birth cohort study
Mengjuan LU ; Kun HUANG ; Shuangqin YAN ; Beibei ZHU ; Shanshan SHAO ; Peng ZHU ; Fangbiao TAO
Chinese Journal of Epidemiology 2020;41(7):1072-1075
Objective:To investigate the impacts of antenatal anxiety on preterm birth and low birth weight.Methods:Women in early pregnancy were recruited for follow-up, antenatal anxiety in three trimesters was screened using Self-Rating Anxiety Scale and the score ≥50 was regarded as anxiety. Logistic regression analysis was conducted to evaluate the associations of the anxiety in three trimesters, new onset anxiety in the second and third trimesters with infant birth outcomes, such as preterm birth and low birth weight.Results:The rates of anxiety in the first, second and third trimesters of pregnancy were 12.5%, 3.7%, and 7.4% respectively. We found that there was no statistical association between anxiety in the first and second trimester and preterm birth. The anxiety in the third trimester was associated with increased odds for preterm birth ( OR=3.55, 95 %CI: 1.62-7.82). Associations between anxiety in all three trimesters and low birth weight were not significant. New onset anxiety in the third trimester was associated with significant increased risk of premature delivery ( OR=5.20, 95 %CI: 1.84-14.70) and low birth weight ( OR=6.93, 95 %CI: 2.42-19.88). Conclusions:Our study showed that anxiety in the third trimester is an important risk factor for premature delivery, new onset anxiety symptoms in the third trimester can significantly increase the incidence of premature birth and low birth weight of infant.
6. Study on the accuracy of oxygen concentration of modified oxygen treatment with Venturi and humidity system
Qiang WEI ; Bingyu QIN ; Guojun HE ; Yuanyuan WU ; Yuan SHI ; Weitao SUN ; Mengjuan JING ; Shichao ZHU ; Huanzhang SHAO
Chinese Critical Care Medicine 2018;30(7):677-680
Objective:
To verify the accuracy of oxygen concentration (FiO2) of modified oxygen treatment with Venturi and humidity system.
Methods:
Patients just after ventilator weaning and before the removal of tracheal intubation/tracheotomy tube, who admitted to the intensive care unit (ICU) of Henan Provincial People's Hospital from May 1st to December 15th in 2017, were enrolled. All patients were given a modified oxygen treatment with Venturi and humidity system, and the oxygen flow rate (Flow) of the Venturi device and the oretical value of FiO2 were adjusted according to the patient's condition. Patients were divided into five groups based on doctor's orders: Flow 3 L/min FiO2 0.24, Flow 3 L/min FiO2 0.26, Flow 6 L/min FiO2 0.28, Flow 6 L/min FiO2 0.30, Flow 9 L/min FiO2 0.35. The value of FiO2 at the inhalation end of patients of each group was measured by TSI airflow analyzer, and the consistency between the measured value of FiO2 at the inhalation end and the FiO2 marked value of Venturi was compared and analyzed.
Results:
When the FiO2 theoretical value of Venturi were adjusted to 0.24, 0.26, 0.28, 0.30, and 0.35, the measured values of FiO2 at the inhalation end of patients were 0.38±0.05, 0.38±0.05, 0.40±0.04, 0.41±0.04, and 0.77±0.11, respectively, which were all significantly higher than the theoretical value of FiO2 (all
7.Study on the accuracy of oxygen concentration of modified oxygen treatment with Venturi and humidity system.
Qiang WEI ; Bingyu QIN ; Guojun HE ; Yuanyuan WU ; Yuan SHI ; Weitao SUN ; Mengjuan JING ; Shichao ZHU ; Huanzhang SHAO
Chinese Critical Care Medicine 2018;30(7):677-680
OBJECTIVE:
To verify the accuracy of oxygen concentration (FiO2) of modified oxygen treatment with Venturi and humidity system.
METHODS:
Patients just after ventilator weaning and before the removal of tracheal intubation/tracheotomy tube, who admitted to the intensive care unit (ICU) of Henan Provincial People's Hospital from May 1st to December 15th in 2017, were enrolled. All patients were given a modified oxygen treatment with Venturi and humidity system, and the oxygen flow rate (Flow) of the Venturi device and the oretical value of FiO2 were adjusted according to the patient's condition. Patients were divided into five groups based on doctor's orders: Flow 3 L/min FiO2 0.24, Flow 3 L/min FiO2 0.26, Flow 6 L/min FiO2 0.28, Flow 6 L/min FiO2 0.30, Flow 9 L/min FiO2 0.35. The value of FiO2 at the inhalation end of patients of each group was measured by TSI airflow analyzer, and the consistency between the measured value of FiO2 at the inhalation end and the FiO2 marked value of Venturi was compared and analyzed.
RESULTS:
When the FiO2 theoretical value of Venturi were adjusted to 0.24, 0.26, 0.28, 0.30, and 0.35, the measured values of FiO2 at the inhalation end of patients were 0.38±0.05, 0.38±0.05, 0.40±0.04, 0.41±0.04, and 0.77±0.11, respectively, which were all significantly higher than the theoretical value of FiO2 (all P < 0.01). The difference between the measured value of FiO2 at the inhalation side and the FiO2 value of the Venturi annotated and the difference rate were both "V"-shaped, both of which decreased with the increase in theoretical value of FiO2 to a Flow of 9 L/min and a theoretical value of FiO2 0.35, the accuracy was the worst, with the FiO2 difference of 0.42±0.11, and the FiO2 difference rate of (121.6±36.5)%.
CONCLUSIONS
There is a difference between the measured value and the theoretical value of FiO2 at the inhalation end of the modified Venturi oxygen therapy humidification system, which needs to be paid attention to during clinical oxygen therapy.
Humans
;
Humidity
;
Oxygen/analysis*
;
Oxygen Inhalation Therapy
;
Respiration, Artificial
;
Ventilator Weaning


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