The signaling pathways involved in acupuncture intervention in post-stroke depression (PSD) mainly include cAMP signaling pathway, MAPK signaling pathway, PI3K/Akt/mTOR signaling pathway, NF-κB signaling pathway, CREB signaling pathway, and CaMK signaling pathway. A variety of regulatory networks are formed between these signaling pathways, which play a key role in reducing inflammation, inhibiting apoptosis, improving neuroplasticity, promoting angiogenesis, and protecting neurons.

Objective To establish an immortalized tree shrew corneal stromal cells(CSCs)line and to study its response to virus infection.Methods Primary tree shrew CSCs were isolated and cultured by the tissue block adhesion method.CSCs were then transfected with a lentivirus carrying the SV40T gene and monoclonal cells were selected for passage culture.The characteristics of the CSCs were investigated by morphological observation and compared with 40 generations until the 50 generations or more,immunofluorescence identification of vimentin and SV40T genes,karyotype examination,and cell proliferation curve.The CSCs were infected with herpes simplex virus-1(HSV-1)(McKrae strain),Zika virus(ZIKV,GZ01 strain),Dengue virus typeⅡ,and H1N1(PR8).Results The immortalized tree shrew CSCs after＞50 passages appeared spindle-shaped with good cell morphology and structure compared with 40 generations.Positive immunofluorescence expression of vimentin and SV40T genes.The cell growth curve showed that the cells were in logarithmic-phase growth on days 4～5 and grew vigorously.The number of chromosomes in the primary cells was stable at 62,while immortalized CSCs had 64 chromosomes at P21 and P56.The virus titer results showed that the immortalized tree shrew CSCs were sensitive to HSV-1(McKrae strain),ZIKV(GZ01 strain),Dengue virus typeⅡ,and H1N1(PR8),with virus titers of 1.32×105,5.62×106,2.69×107,and 7.76×104 CCID50/mL,respectively.Conclusions The immortalized tree shrew CSCs were established successfully,suggesting that this cell line is suitable for studies of the mechanisms of HSV,ZIKV,Dengue virus,and influenza A virus infection in relation to corneal diseases and antiviral drugs.

Influenza is a highly contagious disease that mainly affects the respiratory system and often leads to lung complications. Also it can cause a variety of very rare and serious neurological complications, including Guillain-Barre syndrome, transverse myelitis, meningoencephalitis and others. In recent years, neurological complications caused by influenza A virus have been reported in many countries and regions, and gradually attracted international attention. However, the pathogenesis of this complication remains unclear, and there are few related cases and animal experimental studies,and no specific treatment. Therefore, the authors summarized the study of neurological complications caused by influenza A virus in human and laboratory animals, in order to have a comprehensive understanding of the neurological diseases caused by influenza A virus.

Objective:To investigate the nutritional and metabolic risk factors associated with recurrence in patients with newly diagnosed ulcerative colitis (UC), so as to allow better clinical prediction of recurrence.Methods:A retrospective cohort study was conducted. Patients newly diagnosed with UC (mild and moderate) from the People's Hospital of Xinjiang Uygur Autonomous Region were screened based on prespecified inclusion and exclusion criteria from January 2016 to January 2019. Patients were followed up regularly for three years. Subgroups were determined according to the presence or absence of recurrence. The patients in the UC recurrence group were further stratified according to the time to recurrence into short-term (0-6 months), mid-term (6-12 months) and long-term (12-36 months) recurrence groups. The nutritional and metabolic risk factors related to recurrence were evaluated by univariate analysis and multifactorial logistic regression analysis, and the predictive value was evaluated via receiver operating characteristic curve. The risk factors were then compared across the 3 subgroups with recurrence.Results:A total of 210 patients newly diagnosed with UC (mild and moderate) were included, including 38 experiencing recurrence within 0-6 months, 27 within 6-12 months, 24 within 12-36 months, and 121 without recurrence. There were no statistically significant differences in gender, age, smoking history, and family history in the recurrence group compared with the non-recurrence group. Univariate analysis suggested significant differences in homocysteine, folate, total cholesterol, triglycerides, high-density lipoprotein (HDL), low-density lipoprotein (LDL), apolipoprotein A/B (ApoA/B), 25-hydroxy vitamin D 3, and body mass index (BMI) between recurrence and non-recurrence groups ( P < 0.05). Multifactorial binary logistic regression analysis suggested that homocysteine ( OR = 0.869, 95% CI: 0.782 to 0.965, P = 0.009), triglycerides ( OR = 0.176, 95% CI: 0.060 to 0.519, P = 0.002), LDL ( OR = 0.256, 95% CI: 0.089 to 0.733, P = 0.011), 25-hydroxy vitamin D 3 ( OR = 0.937, 95% CI: 0.895 to 0.0.982, P = 0.006), and BMI ( OR = 1.319, 95% CI: 1.162 to 1.498, P < 0.01) were independent risk factors for UC recurrence. The predictive efficiency of individual risk factors in descending order was as following: LDL (AUC = 0.762, Youden's index [YI] = 0.42, cut-off value = 2.345), triglycerides (AUC = 0.718, YI = 0.361, cut-off value = 1), homocysteine (AUC = 0.666, YI = 0.283, cut-off value = 13.265). There were no statistically significant differences in gender, age, smoking history, and family history across the short-term, mid-term and long-term recurrence groups. There were significant differences in HDL and ApoA/B levels between the short-term and the long-term recurrence groups ( P < 0.05). Conclusions:Recurrence of the disease in UC patients results from the combined effects of multiple factors. The changes in homocysteine, triglycerides, LDL, 25-hydroxy vitamin D 3, and BMI in UC patients should be proactively monitored to prevent recurrence.

The purpose of this study is to provide a culture for mouse bone marrow-derived macrophages (BMDM) and peritoneal macrophages (PM) and to characterize their molecular and cellular biology. The cell number and purity from the primary culture were assessed by cell counter and flow cytometry, respectively. Morphological features were evaluated by inverted microscope. Phagocytosis by macrophages was detected by the neutral red dye uptake assay. Phenotypic markers were analyzed by real-time fluorescent quantitative PCR. Our results show that the cell number was much higher from culture of BMDM than PM, while there was no significant difference regarding the percentage of F4/80+CD11b+ cells (98.30%±0.53% vs. 94.83%±1.42%; P>0.05). The proliferation rate of BMDM was significantly higher than PM in the presence of L929 cell conditioned medium, by using CCK-8 assay. However, PM appeared to adhere to the flask wall and extend earlier than BMDM. The phagocytosis capability of un-stimulated BMDM was significantly higher than PM, as well as lipopolysaccharide (LPS)-stimulated BMDM, except the BMDM stimulated by low dose LPS (0.1 μg/mL). Furthermore, Tnfα expression was significantly higher in un-stimulated BMDM than PM, while Arg1 and Ym1 mRNA expression were significantly lower than PM. The expression difference was persistent if stimulated by LPS+IFN-γ or IL-4. Our data indicate that bone marrow can get larger amounts of macrophages than peritoneal cavity. However, it should be aware that the molecular and cellular characteristics were different between these two culture systems.
Animals ; Bone Marrow Cells ; physiology ; Cells, Cultured ; Culture Media, Conditioned ; Lipopolysaccharides ; metabolism ; Macrophages ; classification ; physiology ; Mice ; Phagocytosis

Objective To explore the characteristics of immunological changes in tree shrews infected with orthoreovirus, and provide a theoretical basis for the prevention of virus in tree shrews. Methods 40 -50-day-old tree shrews were divided into three groups: MRV1/TS/2011 virus-infected and MRV3/TS/2013 virus-infected groups, and saline-treated control group. On the 1, 8, 14, 21, and 28 days after infection, blood samples were taken from the tail vein and used for RT-PCR, flow cytometry and ELISA detection, to assess the viral load, number of CD4/CD8/CD19 cells, and IFN-gamma expression. Results The MRV1/TS/2011 and MRV3/TS/2013 viral load in the plasma and the number of CD4 +and CD19 +cells reached a peak at the 14th day after infection. At the first day after MRV1/TS/2011 infection, the CD4 +cells had a significantly higher expression compared with the normal group. CD8 +cells and the IFN-gamma expression reached a peak at the 21st day after infection. The expression of CD4 +was even higher after MRV1/TS/2011 infection, and the expression of CD8 +cells was higher after MRV3/TS/2013 infection. Conclusions We would conclude that after MRV1/TS/2011 and MRV3/TS/2011 virus infection, accompanying the changes of viral load, it shows some regularity of the expression of CD4/CD8/CD19 and IFN-gamma in the tree shrews: at the early stage of MRV1/TS/2011 virus infection, humoral immunity is stimulated, and CD4 +cells play a major role. MRV3/TS/2013 virus may mainly affect the cellular immunity, while humoral immunity only plays a role at a high viral expression or the late stage of infection. CD4 +cells may be more sensitive to type 1 reovirus, and CD19 +cells may be more sensitive to type 3 reovirus.

Objective To establish a quick and accurate method for detection of tree shrew adenovirus(TAV) using TaqMan real-time fluorescence quantitative PCR. Methods Based on the published TAV genome sequence, a 3' conserved sequence was used to design specific probe primers. A standard curve was prepared using a recombinant plasmid containing the target gene fragment. A real-time fluorescence quantitative PCR method was established for detecting TAV based on TaqMan probe. Results The detection method was specific and was not cross-reactive with other common pathogens. The detection limit of the method was 3.7 copies/μL,showing a high sensitivity. The correlation coefficient was 0.998, and the efficiency was 95.7%. The amplification result showed a fine linear relationship,and the repeatability test effect was good. Conclusions The TAV real-time quantitative PCR detection method based on TaqMan probe has been successfully established. It has high sensitivity and reproducibility and can be applied to early detection of TAV infection.

Parkinson's disease(PD)is a progressive neurodegenerative disorder, with an etiology that is now considered to be due to interaction between genetic and environmental factors. Typical PD features include loss of dopaminergic neurons in the nigrostriatal region, with typical motor traits of PD associated with dopamine deficiency. Animal models have contributed to determining PD etiology and pathogenesis,as well as testing new therapeutic schedules and novel drug research. Rodents, tree shrews, primates, and other animal models of PD have been established by different method. These models each have their own advantages and limitations, showing different clinical features and pathological mechanisms to those in humans. Therefore, the appropriate model for scientific research must be carefully considered. This article reviews the main neurotoxic and transgenic models of PD.

Objective To preliminarily explore the feasibility of tree shrew as a new kind of animal model in research of amblyopia,to discuss the primary visual cortex plasticity mechanism of form deprivation in tree shrew,and to provide a theoretical basis for further understanding the mechanism of amblyopia formation and recovery.Methods Sixty 30-days old tree shrews were divided into five groups,12 in each group:the group A had the right eye sutured for 1 month;the group B had the right eye sutured for 2 months;the group C had the left eye sutured for 1 month and then opened and the righ eye was sutured for 1 month,in other words,the group C was performed by alternating suture;the tree shrews of control group 1(D1) were in the same age as the the group A,but fed in normal breedingenvironment;the tree shrews of control group 2(D2) were at the same age of groups B and C,but fed with a normal diet.Samples of the visual cortex were taken after the completion of modeling,and were processed to observe the histology and ultrastructure of the visual cortex,the neuron apoptosis,and the c-fos protein expression in the tree shrews of different groups.Results Damages to different degrees were found by histological and electron microscopic examination of the visual cortex in each experimental group,and they were more obvious in the group sutured for 2 months.TUNEL staining showed that there were no significant differences between the apoptosis in the experimental and control groups.The expression of c-fos mRNA and protein in the experimental groups was decreased,and it was the lowest in the group sutured for 2 months.There was a small increase in the c-fos expression after the alternate suture,and no significant difference of c-fos expression was found in the control groups.Conclusions Different degrees of deprivation amblyopia lead to different histopathological changes.There is a plasticity in the neurons affected by amblyopia.Tree shrew can be used as an ideal animal model for the studies of form deprivation amblyopia.

Objective To establish an enterovirus 71(EV71) infection model of tree shrew primary renal cells.Methods Tree shrew primary renal cells were obtained by trypsin digestion.After subculture and purification,EV71 virus was used to infect these primary cells.The culture supernatant of these EV71-infected cells was collected for virus titer detection at 1,2,4,6 and 8 days post-infection.The cells were collected for detection of EV71 VP1 protein by Western blot assay.Furthermore,the expression and location of VP1 protein in the infected cells were detected by indirect immunofluorescence assay.Vero cells were taken as positive control to evaluate the infectivity of EV71 virus to tree shrew primary renal cells.Results Morphologically,the cultured cells were proved to be majorly consisted of the primary renal cells after subculture and purification.The obtained primary cells were infected by EV71 virus.The virus titer was up to 1.3×106 TCID 50/mL during 48-96 h post-infection,proving that EV71 virus infected and proliferated in the tree shrew primary renal cells.Western blot showed that the viral VP1 protein was detected from infected primary cells at 2 to 8 d post infection.VP1 protein was also observed in the cytoplasm at 2 to 6 d post infection by indirect immunofluorescence.Compared with Vero cells,the infectivity of EV71 virus to tree shrew primary renal cells and its proliferation were confirmed.Conclusions Based on the successful establishment of cell culture of tree shrew primary renal cells,the infectivity to the obtained cells and proliferation of EV71 virus in the cells are confirmed.The model of EV71 virus-infected tree shrew primary renal cells is initially established.