Objective To explore the isolation, culture and identification of mouse amniotic fluid-derived mesenchymal stem cell (AF-MSC). Methods The uteruses of pregnant mice were obtained under sterile conditions. The amniotic fluid was collected, filtered and centrifuged, and the precipitated cell mass was cultured and passaged. The morphology of AF-MSC was observed and the proliferation characteristics of AF-MSC were analyzed. The surface markers of AF-MSC were identified by flow cytometry. The osteogenic, chondrogenic and adipogenic differentiation capability of AF-MSC and cell vitality after cryopreservation and resuscitation were evaluated. Results The mouse AF-MSC was seen in typical spindle shape, and vortex structure could be observed when the cell confluency exceeded 80%. No evident latency was noted in the passage and culture of mouse AF-MSC. After 2-3 d culture, AF-MSC proliferated in the logarithmic growth stage with the fastest growth rate, which was slowed down and entered into the plateau period. AF-MSC expressed stem cell antigen (Sca)-1, CD29 and CD44 rather than CD34 and CD45. After the osteogenic differentiation of mouse AF-MSC, the mineralized crystals were stained in dark red spots by Alizarin red S staining. After chondrogenic differentiation, the secreted acid mucopolysaccharide was stained in light blue by Alcian blue. After adipogenic differentiation, cytoplasmic lipid droplets were stained in red by oil red O staining. After cryopreservation and resuscitation, the survival rate of AF-MSC exceeded 95%, and the growth status was excellent. The proliferation ability at 6 d was significantly better than that before cryopreservation (P < 0.05), and the proliferation ability at other time points did not significantly differ from that before cryopreservation (all P > 0.05). Conclusions Mouse AF-MSC may be successfully isolated with convenient procedure and the low cost. In addition, the isolated AF-MSC may be purified along with the increasing times of passage. Cryopreservation does not affect the proliferation ability of AF-MSC.

Migraine is the most common type of primary headache with disabling brain dysfunction. The prevalence of the migraine in Chinese population is 9.3%, and the ratio of female to male is 3∶1, which seriously affect people′s quality of life. Depression is one of the most common psychiatric disorders in migraine comorbidity. Compared with non-migraine patients, the risk of depression comorbidity in migraine patients is more than 2.5 times higher. The frequency and severity of migraine are closely related to depressive symptoms. Depressive symptoms have different effects on headache-related pain signal transduction, which is susceptible to neuroendocrine network disorders in the process of transmission from thalamus to cortex. Neuroendocrine network plays an important role in the depression of migraine comorbidity. Therefore, exploring the pathogenesis of neuroendocrine network in comorbidity provides a theoretical basis for screening more suitable depressive drugs for migraine comorbidity.

Objective To investigate the association between the polymorphism of the microsomal triglyceride transport protein (MTTP) gene at rs1800591 locus and the risk of nonalcoholic fatty liver disease (NAFLD) in the elderly population. Methods The clinical cohort of this study was established in Menkuang Hospital, Beijing Jingmei Group General Hospital. A total of 1098 healthy elderly volunteers were recruited for physical examination in communities in Mentougou District of Beijing, China, from January 11, 2020 to September 30, 2021, among whom there were 614 patients with NAFLD and 484 individuals without NAFLD. Gene microarray was used to determine the genotypes of MTTP rs1800591; demographic data were collected, and blood biochemical parameters were measured. The independent samples t -test was used for comparison of normally distributed continuous data between groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups; the chi-square test was used for comparison of categorical data between groups. The chi-square test was used to investigate whether the distribution of genotype frequency was in accordance with Hardy-Weinberg equilibrium. The unconditional logistic regression model was used to calculate odds ratio ( OR ) and its 95% confidence interval ( CI ) to investigate the association of gene polymorphism with the risk of NAFLD and other comorbidities. Results There were significant differences in sex and age between the two groups ( P < 0.05). Compared with the non-NAFLD group, the NAFLD group had significantly higher levels of body mass index (BMI), waist-hip ratio, triglyceride, alanine aminotransferase, aspartate aminotransferase, controlled attenuation parameter (CAP), and liver stiffness measurement and a significantly lower level of high-density lipoprotein (HDL) (all P < 0.05). Compared with the non-NAFLD group, the NAFLD group had a significantly higher proportion of patients with hypertension, diabetes, obesity, and metabolic syndrome (all P < 0.05). The distribution of genotype frequency at MTTP rs1800591 locus was in accordance with Hardy-Weinberg equilibrium in the control group ( χ 2 =1.097, P =0.29). There were a significant differences in the genotype and the distribution of alleles at MTTP rs1800591 locus between the patients with NAFLD and the control group (all P < 0.001). In the total population, there was a significantly lower carrying rate of T allele (GT+TT, n =351) in male individuals, and the individuals carrying T allele had significantly higher BMI and CAP than those carrying GG allele ( n =747) ( P < 0.001). Compared with the individuals who did not carry T allele, the individuals carrying T allele (GT+TT, n =232) had a significantly higher proportion of patients with obesity and a significantly lower NFS score ( P < 0.05). As for the individuals with NAFLD, the individuals carrying T allele had a significantly lower proportion of male individuals, a significantly lower waist-hip ratio, and a significantly higher level of HDL compared with those who did not carry T allele (GG, n =382), and the GT+TT group had a significantly lower NFS score than the GG group (all P < 0.05). The non-conditional logistic regression analysis showed that after adjustment for the confounding factors of sex, age, and BMI, the GT+TT genotype at MTTP rs1800591 locus significantly increased the risk of NAFLD ( OR =1.643, 95% CI : 1.226-2.203, P =0.001), and carrying T allele also increased the risk of obesity in the total population ( OR =1.371, 95% CI : 1.051-1.788, P =0.02). Conclusion MTTP rs1800591 polymorphism is associated with the development of NAFLD in the elderly population, and carrying T allele may promote hepatic steatosis and increase the risk of obesity in NAFLD, while it may inhibit the progression of liver fibrosis.