1.Effect of Mahuang Xixin Fuzitang on Migration of Dendritic Cells in Mice by Regulating Rho/ROCK Signaling Pathway
Shuyue FAN ; Luwei WU ; Tongtong SUN ; Yuying NI ; Jing GAO ; Ying CUI ; Min LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):35-42
ObjectiveTo investigate the inhibitory effect of Mahuang Xixin Fuzitang on the migration of dendritic cells (DCs) in mice and its underlying mechanism. MethodMouse bone marrow-derived DCs were isolated and cultured. The morphological changes of the cells at different stages were observed under a microscope, and the CD11c+ proportion was detected by flow cytometry to identify DC purity. Cells were treated with Mahuang Xixin Fuzitang (1, 2, 5, 10, 20, 40, 50, 100 g·L-1) for 24 hours, and the effect of Mahuang Xixin Fuzitang on cell proliferation was assessed using the cell counting kit-8 (CCK-8) assay to determine the appropriate concentrations for treatment. After modeling by lipopolysaccharide (LPS) induction, DCs were divided into a blank group, a model group, and Mahuang Xixin Fuzitang groups (2, 4, 8 g·L-1). The expression of surface molecules CD80, CD86, and major histocompatibility complex-Ⅱ (MHC-Ⅱ) were detected by flow cytometry. Transwell chamber assay was used to observe cell migration. The levels of chemokine C-C-primitive receptor 7 (CCR7) and chemokine C-X-C-primitive receptor 4 (CXCR4) on the cell surface were detected by enzyme-linked immunosorbent assay (ELISA). The expression of filamentous actin (F-actin) in the cell microfilament cytoskeleton was detected by immunofluorescence (IF) staining. Real-time quantitative polymerase chain reaction (Real-time PCR) was employed to determine the mRNA expression levels of Ras homolog family member A (RhoA) and Rho-associated coiled-coil containing protein kinase 1 (ROCK1). Western blot analysis was performed to detect the protein expression of RhoA and ROCK1. ResultCompared with the blank group, the model group exhibited significantly higher expression levels of CD80, CD86, and MHC-Ⅱ (P<0.01), a significantly increased number of cells migrating to the lower chamber (P<0.01), and significantly elevated levels of CCR7 and CXCR4 (P<0.05, P<0.01). Additionally, F-actin expression was significantly increased (P<0.01), and both RhoA and ROCK1 mRNA and protein expression levels were significantly higher (P<0.05, P<0.01). Compared with the model group, treatment with Mahuang Xixin Fuzitang (2, 4, 8 g·L-1) for 24 hours resulted in significantly lower expression levels of CD80, CD86, and MHC-Ⅱ (P<0.01), a significantly reduced number of cells migrating to the lower chamber (P<0.05), and significantly decreased levels of CCR7 and CXCR4 (P<0.05, P<0.01). Furthermore, F-actin expression was significantly reduced (P<0.01), and both RhoA and ROCK1 mRNA and protein expression levels were significantly decreased (P<0.05, P<0.01). ConclusionMahuang Xixin Fuzitang can inhibit the migration of DCs in mice, and its mechanism of action may be related to reducing the activity of the Rho/ROCK signaling pathway, thereby affecting changes in the cell cytoskeleton.
2.Quantitative research on China's disability rehabilitation policy using policy modeling consistency index model
Tongtong GUO ; Xinyi ZHANG ; Lihong JI ; Zhiwei DONG ; Zongrun LI ; Liduan WANG ; Weiqin CAI ; Qianqian GAO ; Qi JING ; Wengui ZHENG
Chinese Journal of Rehabilitation Theory and Practice 2024;30(6):621-629
Objective To quantitatively analyze and evaluate the content of rehabilitation policy for people with disabilities in China. Methods This study focused on ten national policies of disability rehabilitation issued from 2021 to 2023.It employed text mining techniques to process policy texts and constructed a policy modeling consistency index model for dis-ability rehabilitation policies in China.The relevant policies were evaluated and analyzed quantitatively. Results The disability rehabilitation policies in China were relatively comprehensive in terms of policy transparency,op-erational mechanisms and policy nature.However,there was still a need for optimization in terms of policy per-spectives,target groups,incentive mechanisms,and other aspects. Conclusion The overall quality of disability rehabilitation policy texts at the national level in China is relatively good.There is a need to further enhance the predictability of policy objectives,clarify the responsibilities and division of labor among various departments,and improve policy incentive mechanisms in future policy formulation,which will further refine China's disability rehabilitation policy system and contribute to high-quality develop-ment of the disability cause.
3.Effects of sinomenine hydrochloride on ultrastructure and STING ex-pression in kidney tissues of db/db mice
Tongtong HE ; Xiaofei JIN ; Chunyue ZUO ; Xiaohong ZHOU ; Yu GAO ; Xiangmei CHEN ; Weijuan GAO
Chinese Journal of Pathophysiology 2024;40(11):2074-2080
AIM:This study aimed to investigate the effects of sinomenine hydrochloride(SIN)on the ultra-structure of renal tissue and the expression of interferon gene-stimulating factor in db/db mice.METHODS:Sixteen 12-week-old male db/db mice were randomly divided into two groups:a model group and a sinomenine hydrochloride(SIN)group,each consisting of 8 mice.An additional 8 wild-type(WT)mice served as the normal control group.The sinome-nine hydrochloride group was administered the treatment for 8 weeks,followed by a 20-week observation period,while the normal and model groups received an equal volume of saline via gavage.Weekly measurements were taken for body weight and fasting blood glucose.Serum creatinine(SCr)and blood urea nitrogen(BUN)levels were assessed,and 24-hour uri-nary microalbumin(ALB)levels,as well as serum inflammatory cytokines interleukin-1β(IL-1β),IL-6 and tumor necro-sis factor-α(TNF-α),were determined using ELISA.Pathological changes in renal tissue were evaluated through hema-toxylin-eosin(HE)staining,while ultrastructural alterations were examined using transmission electron microscopy.Im-munohistochemistry and Western blotting were employed to assess STING protein expression in renal tissue,and STING mRNA expression was quantified via RT-qPCR.RESULTS:Compared to the normal group,the model group exhibited significant increases in BUN,ALB,and SCr levels(P<0.01),alongside elevated inflammatory markers IL-1β,IL-6,and TNF-α(P<0.01).Notable pathological changes included leukocyte wall thickening in capillaries,inflammatory cell infiltration,increased mesangial matrix,disorganized and linear alignment of podocytes,and thickening of the basement membrane.Moreover,STING protein and mRNA expression levels were significantly elevated(P<0.01).In contrast,the sinomenine hydrochloride group demonstrated significantly reduced levels of renal function markers(BUN,ALB and SCr)compared to the model group(P<0.01),as well as decreased concentrations of inflammatory factors IL-1β,IL-6,and TNF-α(P<0.01).Improvements in renal histopathology included decreased leukocyte wall thickening,reduced inflam-matory cell presence,diminished mesangial matrix,and a significant reduction in foot process fusion,alongside thinner basement membranes.Both STING protein and mRNA expression levels were also significantly lower(P<0.01).CON-CLUSION:Sinomenine hydrochloride effectively mitigates renal tissue injury,improves ultrastructural alterations,and inhibits inflammatory responses in db/db mice.Its mechanism of action appears closely linked to the downregulation of STING protein and mRNA expression.
4.Quantification of antigen of Mycoplasma capricolum subsp. capripneumoniae by optical assay.
Jiazhen GE ; Pengcheng GAO ; Tongtong TIAN ; Xiaoni WU ; Qianqian LI ; Kexin TIAN ; Guodong SONG ; Fuying ZHENG ; Yuefeng CHU
Chinese Journal of Biotechnology 2023;39(12):4874-4886
Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the cause of contagious caprine pleuropneumonia (CCPP) in goats. Inactivated vaccines and capsular polysaccharide (CPS) indirect hemagglutination reagents are available for prevention and serological detection, but high culture costs and complex antigen quantification have been plagued by production staff. In order to solve these problems in production practice, a sugar fermentation medium with an initial pH value of 7.8, which could improve the production of two antigens simultaneously, was screened out by changing the initial pH value based on previous Mccp metabolomics analysis. Since phenol red can be identified by UV absorption spectrum and cetyltrimethylammonium bromide (CTAB) can bind to anionic capsular polysaccharide, a UV spectrum measurement method for analyzing the culture stage reached by Mccp and a CTAB precipitation test for relative quantification of capsular polysaccharide antigen content in the fermentation broth were established. The UV spectrum observation method can guide the production of Mccp according to the growth curve of Mccp, which greatly reduces the monitoring time of the traditional CCU method and improves the accuracy of the original eye-observation method. The established CTAB precipitation test can complete the monitoring of CPS content within 5 hours, which greatly reduces the time required compared with the traditional differential technique, and its accuracy was verified by the phenol-sulfuric acid method. The optimized culture medium and the two correlation comparison methods established in this study can effectively reduce the production cost of Mccp and improve the production efficiency. The two assays have been used in the research at our laboratory, which provides experimental data for further improvement of the production process of CCPP inactivated vaccine and capsular polysaccharide as well as rapid quantification.
Humans
;
Animals
;
Goats
;
Cetrimonium
;
Mycoplasma
;
Polysaccharides
5.Influence of copy number variations in the FCGR3A and FCGR3B genes on the outcome of hepatitis B virus infection
Haotian LI ; Tongtong WANG ; Xutong LI ; Yufeng GAO
Journal of Clinical Hepatology 2022;38(6):1275-1279
Objective To investigate the association of copy number variations (CNVs) in the FCGR3A and FCGR3B genes with different outcomes and disease progression after hepatitis B virus (HBV) infection. Methods Peripheral blood samples were collected from 841 patients with chronic HBV infection and 296 patients with self-limited HBV infection, an according to the degree of disease progression, the patients with chronic HBV infection were further divided into chronic hepatitis B (CHB) group, liver cirrhosis (LC) group, and hepatocellular carcinoma (HCC) group. The AccuCopy technique was used for the quantitative analysis of CNVs in the FCGR3A and FCGR3B genes in peripheral blood. The independent samples t -test was used for comparison of continuous data between two groups, and a one-way analysis of variance and the Kruskal-Wallis H test were used for comparison between multiple groups; the chi-square test was used for comparison of categorical data between groups. The chi-square test was also used to investigate the difference in the distribution of CNVs in the FCGR3 gene between different groups. The age-and sex-adjusted logistic regression model was used to investigate the influence of CNVs on the chronicity of HBV infection. Results There was a significant difference in the frequency distribution of CNVs in the FCGR3A and FCGR3B genes between the chronic HBV infection group and the self-limited HBV infection group ( χ 2 =11.406 and 19.143, both P < 0.05). As for disease progression after chronic HBV infection, there were no significant differences in CNVs of the FCGR3A and FCGR3B genes between the CHB group, the LC group, and the HCC group (FCGR3A: χ 2 =3.125, P =0.537; FCGR3B: χ 2 =5.274, P =0.260). There were also no significant differences in CNVs of the FCGR3A and FCGR3B genes between the HBeAg-positive group and the HBeAg-negative group (FCGR3A: χ 2 =1.025, P =0.599; FCGR3B: χ 2 =0.712, P =0.701). Reduction or deletion of the copy number of the FCGR3A and FCGR3B genes was a risk factor for the chronicity of HBV infection (FCGR3A: odds ratio [ OR ]=0.621, 95% confidence interval [ CI ]: 0.513-0.752; FCGR3B: OR =0.594, 95% CI : 0.491-0.719). Conclusion Reduction or deletion of the copy number of the FCGR3A and FCGR3B genes may be a genetic susceptibility factor for the chronicity of HBV infection, but it is not associated with disease progression.
6.Normal reference and regional variation of spinal bone mineral density under the quantitative CT in Chinese male population
Liuping CHEN ; Jian ZHAI ; Limei RAN ; Yongli LI ; Lü YINGRU ; Yan WU ; Shaolin LI ; Hanqi WANG ; Yaling PAN ; Tongtong CHEN ; Lü HAIYING ; Kaiping ZHAO ; Yuqin ZHANG ; Xiao MA ; Jing LU ; Xigang XIAO ; Xiangyang GONG ; Zehong YANG ; Wei CHEN ; Jianbo GAO ; Yuehua LI ; Xia DU ; Zhenlin LI ; Qiang ZENG ; Xiaoguang CHENG ; Shuang CHEN ; Jing WU ; Yong LU
Chinese Journal of Health Management 2022;16(9):604-609
Objective:To investigate the normal reference values of spinal bone mineral density measured by quantitative computed tomography (QCT) and the differences of bone mineral density (BMD) in different regions of in Chinese adult males.Methods:Men who underwent low-dose CT lung scan for cancer screening in regions of Northeast, North, East, South, Central and Southwest of China from January 2018 to December 2019 were selected. And the lumbar vertebrae BMD values in the male subjects were measured by the QCT system (Mindways Software, Inc.). The mean BMD values and their decline rates were calculated at an age interval of 10 years, and the prevalence of osteoporosis was calculated according to the American College of Radiology spine QCT osteoporosis diagnostic criteria.Results:A total of 50 682 males with a mean age of (50.22±12.79) years (ranged 20 to 98 years) were included in this study. The peak BMD of (173.11±28.56) mg/cm 3 in the healthy Chinese adult male population appeared in the age group of 20 to 29 years and then declined with age. Before the age of 70 years, the BMD was relatively higher in males in South China, and it was lower in Central China and Southwest China, and it was intermediate in Northeast, North and East of China, with statistically significant differences. There was no significant differences in BMD in the males in the two age groups of 70 to 79 years and 80 and older among the regions in China. The overall decline rate of spinal BMD in Chinese males under QCT was about 46.92% over the lifetime, and it declined obviouslyin the 40-49 age group. The overall prevalence of osteoporosis in Chinese male population aged 50 years and above was approximately 11.42%, with the highest prevalence in Southwest China and Central China (14.72% and 13.87%, respectively) and the lowest in North China and South China (8.53% and 7.71%, respectively). Conclusions:A reference of lumbar spine BMD values for healthy males in China based on QCT is established. BMD values were highest in South China and Lowest in Central China.
7.Practice of international talents training project for hospitals
Yan HUANG ; Yan ZHANG ; Jing ZHANG ; Pengzhan ZHU ; Rong SUN ; Xin DONG ; Tongtong LIU ; Xiaoyi GAO ; Shaoliang SHI ; Jingjing HE
Chinese Journal of Hospital Administration 2021;37(5):405-407
Talents constitute key resources for the development of healthcare sector, and studying abroad is an important and effective means for their training. Based on the analysis of the current status of China′s international talents training project of hospitals, this study summarized existing problems in such training. The authors covered such six aspects as lawful standardization of project management methods, selection of trainees by levels and by types, innovation of the process tracking management mechanisms and strengthened full-process assessment, all dimensional institutional support, consolidation of the responsibilities by individual disciplines in talent team building, and measures to cope with the impact of COVID-19. In accordance with the research results, the study analyzed the exploration and practice of the international training project under the " Elite Talents Cultivation Project" of the Affiliated Hospital of Qingdao University, and raised targeted recommendations on attention to the strategic and forward-looking planning, precise setup of training goal categories, classified evaluation of study results, and lawful management among others.
8.Risk stratification and treatment progress of smoldering multiple myeloma
Zeyu LIN ; Guangzhong YANG ; Tongtong WANG ; Wenming CHEN ; Wen GAO
Journal of Leukemia & Lymphoma 2021;30(10):626-629
Smoldering multiple myeloma is a kind of heterogeneous asymptomatic plasma cell disease. Some patients have a high risk of developing symptomatic multiple myeloma. However, the starting point and options of treatment for smoldering multiple myeloma patients are still unclear. This article reviews the risk stratification and treatment progress of smoldering multiple myeloma.
9.Establishment of a gp120 transgenic mouse model with 7 nAChR knockout.
Tongtong HU ; Zelong GONG ; Yu WAN ; Yubin LI ; Xuefeng GAO ; Jingxian LUN ; Shenghe HUANG ; Hong CAO
Journal of Southern Medical University 2020;40(8):1184-1191
OBJECTIVE:
To construct a HIV-1 gp120 transgenic mouse model (gp120) with 7 nicotinic acetylcholine receptor (7nAChR) gene knockout.
METHODS:
The 7nAChR gene knockout mice (7R) were crossed with HIV-1gp120 transgenic mice (gp120) to generate F1 generation mice. We selected the F1 mice with the genotype of 7R/gp120 to mate to obtain the F2 mice. The genotypes of the F3 mice were identified by PCR, and the protein expressions in the double transgenic animal model was analyzed by immunohistochemistry. BV2 cells were treated with gp120 protein and 7nAChR inhibitor, and the expressions of IL-1β and TNF- were detected using ELISA.
RESULTS:
The results of PCR showed the bands of the expected size in F3 mice. Two F3 mice with successful double gene editing (7R/gp120) were obtained, and immunohistochemistry showed that the brain tissue of the mice did not express 7 nAChR but with high gp120 protein expression. In the cell experiment, treatment with gp120 promoted the secretion of IL-1β and TNF- in BV2 cells, while inhibition of 7nAChR significantly decreased the expression of IL-1β and TNF- ( < 0.001).
CONCLUSIONS
By mating gp120 Tg mice with 7R mice, we obtained gp120 transgenic mice with 7nAChR gene deletion, which serve as a new animal model for exploring the role of 7nAChR in gp120-induced neurotoxicity.
Animals
;
Disease Models, Animal
;
Glycoproteins
;
Mice
;
Mice, Knockout
;
Mice, Transgenic
;
Tumor Necrosis Factor-alpha
;
alpha7 Nicotinic Acetylcholine Receptor
;
metabolism
10.Association of blood lipids and albumin-bilirubin index with disease progression in patients with chronic hepatitis B virus infection
Wenjun CAO ; Tongtong WANG ; Yufeng GAO
Journal of Clinical Hepatology 2019;35(3):522-525
ObjectiveTo investigate the value of blood lipid indices and albumin-bilirubin index (ALBI) in evaluating the progression of chronic hepatitis B virus (HBV) infection. MethodsA total of 184 patients with chronic HBV infection who visited The Second Affiliated Hospital of Anhui Medical University from June 2016 to June 2017 were enrolled, and according to the stage of the disease, they were divided into ASC group (74 HBV carriers), CHB group (70 patients with chronic hepatitis B), and LC group (40 patients with compensated cirrhosis). A total of 50 healthy individuals were enrolled as health control (HC) group. Blood lipid indices and liver function parameters were measured, and the changes in blood lipid indices and ALBI during the progression of chronic HBV infection were analyzed. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the Dunnett method was used for further comparison between two groups; the chi-square test was used for comparison of categorical data between groups; Pearson correlation analysis was used to investigate correlation. Results There were significant differences between the ASC, CHB, LC, and HC groups in blood lipid indices of cholesterol (CHO), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), apolipoprotein A (APOA), and apolipoprotein B (F=12.075, 19.559, 6.554, 9.392, and 5.458, all P<0.001), and the LC group had significantly greater reductions in the above indices compared with the other three groups (all P<0.05). There was a significant difference in ALBI between the four groups (F=49.225, P<0.001); the LC group had a significantly higher ALBI than the other three groups (all P<005), and the ASC and CHB groups had a significantly higher ALBI than the HC group (both P<0.05). CHO, HDL-C, LDL-C, and APOA were negatively correlated with ALBI (all P<0.05), among which CHO and HDL-C had the strongest correlation with ALBI (r=-0.310 and -0.266, both P<0.001). ConclusionIn patients with chronic HBV infection, blood lipid indices and ALBI can reflect the degree of liver function damage, especially in patients with liver cirrhosis.

Result Analysis
Print
Save
E-mail