1.Astragalin Regulates Autophagy and Apoptosis of Astrocytes in L4-5 Spinal Dorsal Horn of Mouse Inflammatory Pain Model
Weishan ZHANG ; Jiahong LIN ; Can WANG ; Runheng ZHANG ; Junhua YANG ; Jing LIU ; Guoying LI ; Yuxin MA
Journal of Sun Yat-sen University(Medical Sciences) 2025;46(2):186-196
ObjectiveTo explore the effects of astragalin (AST) on autophagy and apoptosis of astrocytes in the L4-5 dorsal horn of the spinal cord in mice with inflammatory pain induced by complete Freund's adjuvant (CFA). MethodsTwenty-four male C57BL/6 mice, aged six months, were randomly assigned to four groups: control group, saline group, CFA model group, and CFA+AST group, six mice in each group. The inflammatory pain model was established by injection of 10 µL CFA into the right lateral malleolus fossa. The saline group were injected with an equal amount of normal saline at the same site. The inflammatory pain mice in CFA+AST group were further treated with AST (60 mg/kg) intraperitoneally once a day for 21 consecutive days. Multiplex immunofluorescence staining was used to detect the coexpression of autophagy-related factors including ATG 12 and Beclin-1, apoptosis-related factors including Cleaved-Caspase3 and Caspase9, and the astrocyte marker such as GFAP in the L4-5 spinal dorsal horn of the mice in each group. Western blot was used to examine the protein expression levels of autophagy-related proteins(ATG12, Beclin-1) and apoptosis-related proteins(Caspase 3, Caspase 9) in the L4-5 spinal dorsal horn of mice. ResultsImmunofluorescent staining showed that in the L4-5 dorsal horn of the spinal cord, the fluorescence intensity of ATG12 (P<0.000 1) and Beclin-1 (P<0.000 1) was significantly increased, while that of Cleaved-Caspase 3 (P<0.001) and Caspase 9 (P<0.000 1) was decreased in the CFA+AST group when compared to the CFA model group. Furthermore, AST could inhibit the activation of astrocytes. Western blot further confirmed that AST significantly upregulated the expression of ATG12 (P<0.000 1) and Beclin-1 (P<0.000 1) in the L4-5 spinal cord of CFA mice, and downregulated the expression of Caspase 3 (P<0.01) and Caspase 9 (P<0.001). ConclusionsAST promotes autophagy of astrocytes and inhibits their apoptosis in the L4-5 spinal dorsal horn of CFA mice.
2.Clinical characteristics of 527 cases with different HCV genotypes in Qinghai Province
Shanghai Journal of Preventive Medicine 2025;37(8):682-686
ObjectiveTo investigate the distribution patterns of hepatitis C virus (HCV) genotypes and their clinical characteristics in Qinghai Province, and to provide a theoretical basis for the elimination of hepatitis C in this region. MethodsA total of 527 hepatitis C patients from Qinghai Provincial Infectious Diseases Hospital from August 2023 to August 2024 were selected as the research subjects. Polymerase chain reaction (PCR) sequencing was used to determine the HCV genotypes of the hepatitis C patients, and the distribution patterns of HCV genotypes and the clinical characteristics of different genotypes in this region were observed. ResultsThe 527 hepatitis C patients were mainly distributed in Xining City and Haidong City, with the majority being male patients aged between 48 and <60 years. A total of 4 genotypes and7 subtypes were found, with genotype 2 being the most prevalent one (219 cases, 41.56%), followed by genotype 3 (173 cases, 32.83%),genotype 1 (131 cases, 24.86%) and genotype 6 (6 cases, 0.76%). There were statistically significant differences in gender, age, disease progression, population classification, residential address, transmission route, and liver inflammation indicators among different HCV subtypes (all P<0.05). Genotype 1 and genotype 2 accounted for a higher proportion in chronic hepatitis C patients under 40 years old, females, and transmitted through blood transfusions and invasive procedures, while genotype 3 were more common in male patients, aged ≥40 years old, those infected through intravenous drug use, and those were prone to progress to liver cirrhosis (mainly subtype 3b). Farmers, houseworkers and unemployed people were the main population groups in all genotypes. Compared with other genotypes, genotype 3 had higher levels of aspartate aminotransferase and alpha-fetoprotein and lower platelet counts, suggesting that the severity of the disease was related to different genotypes. ConclusionGenotype 2 and genotype 3 are the main types in Qinghai Province. The composition ratio of genotype 3, which is mainly transmitted through intravenous drug use, is increasing and more likely to progress to liver cirrhosis. This highlights the need to focus on the prevention, treatment and management of genotype 3.
3.Total Saponins of Dioscorea Mitigate Nonalcoholic Steatohepatitis in Mice by Regulating AMPK/SREBP-1c/ACC Signaling Pathway
Yuyan LIU ; Dandan WANG ; Xin WANG ; Guoying LI ; Guangliang CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):41-48
ObjectiveTo investigate the role and mechanism of total saponins of Dioscorea (TSD) in mitigating nonalcoholic steatohepatitis (NASH) in mice. MethodForty-eight C57BL/6J mice were randomized into a normal group and a modeling group. The mice for modeling were fed with a high-fat and high-cholesterol diet + 20% fructose solution for 16 weeks and randomized into model, atorvastatin (4 mg·kg-1·d-1), and high-, medium-, and low-dose (200, 60, and 20 mg·kg-1·d-1) TSD groups. The mice were administrated with corresponding doses of drugs by gavage for 8 weeks. The mouse activity, liver index, levels of total cholesterol (TC), triglycerides (TG), and free fatty acids (FFAs) in the liver, and levels of TC, TG, aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyl transferase (GGT), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in the serum were measured. Hematoxylin-eosin staining, Masson staining, oil red O staining, and transmission electron microscopy were employed to observe the pathological changes, lipid accumulation, and morphological changes of liver ultrastructure. Western blot was employed to determine the protein levels of AMP-activated protein kinase (AMPK), sterol regulatory element-binding protein-1c (SREBP-1c), acetyl-CoA carboxylase (ACC), and phosphorylated ACC (p-ACC) in the liver tissue. ResultCompared with the normal group, the activity of mice in the model group decreased(P<0.05, P<0.01), the levels of TC, TG, FFA and serum TC, TG, ALT, AST, GGT, IL-1β and TNF-α, liver coefficient and liver pathology scores were significantly increased, the expression of p-AMPK/AMPK and p-ACC proteins in liver tissues was significantly reduced, and the expressions of SREBP-1c and ACC proteins were significantly increased (P<0.01). Compared with the model group, atorvastatin increased the mouse activity (P<0.05), while each dose of TSD caused no significant changed in the mouse activity. The levels of TC, TG, FFA in liver and serum TC, TG, ALT, AST, GGT, IL-1β, TNF-α, liver coefficient and liver pathological score in TSD and atorvastatin groups were significantly decreased, and the expressions of p-AMPK/AMPK and p-ACC in liver tissue were significantly increased. The expressions of SREBP-1c and ACC were significantly decreased (P<0.05,P<0.01). ConclusionTSD may alleviate NASH in mice by regulating the AMPK/SREBP-1c/ACC signaling pathway to reduce lipid synthesis.
4.Awareness of air pollution protection knowledge among primary school students in Shennongjia Forest District
LI Xiang ; ZHANG Xiuqin ; LI Guoying ; WANG Qiong ; XIE Shuguang ; FAN Chuangang
Journal of Preventive Medicine 2024;36(2):173-176
Objective:
To investigate the awareness of air pollution protection knowledge and its influencing factors among primary school students in Shennongjia Forest District, Hubei Province, so as to provide insights into targeting implementation of health education on air pollution protection knowledge.
Methods:
Students in Grade 3 to 5 in Shennongjia Shiyan primary school were enrolled by stratified cluster sampling method, and students' demographic features and awareness of air pollution protection knowledge were investigated using the Investigation on the Effects of Air Pollution Health Protection of Pupils (Volume A). Factors affecting the awareness of air pollution protection knowledge among primary school students were identified using a multivariable logistic regression model.
Results:
A total of 897 questionnaires were allocated, and 877 valid questionnaires were recovered, with an effective rate of 97.77%. The respondents included 446 men (50.86%) and 431 women (49.14%), 301 third grade students (34.32%), 284 fourth grade students (32.38%), and 292 fifth grade students (33.30%), and had a mean age of (10.32±0.93) years. The overall awareness of air pollution protection was 55.76%, and the awareness rates of basic concepts, basic knowledge, and basic behaviors and skills were 42.99%, 53.48% and 57.24%, respectively. Multivariable logistic regression analysis identified age (OR=1.453, 95%CI: 1.053-2.005), living with parents (OR=2.638, 95%CI: 1.571-4.429), mother's educational level (below primary school, OR=0.270, 95%CI: 0.084-0.862; primary school, OR=0.169, 95%CI: 0.069-0.416; junior high school, OR=0.309, 95%CI: 0.138-0.691; high school, OR=0.352, 95%CI: 0.160-0.773) and average annual family income (50 000 to 100 000 Yuan, OR=1.629, 95%CI: 1.162-2.282; 100 000 to 150 000 Yuan, OR=1.802, 95%CI: 1.101-2.948; ≥150 000 Yuan, OR=1.939, 95%CI: 1.065-3.529) as factors affecting the awareness of air pollution protection knowledge among primary school students.
Conclusion
The awareness of air pollution protection knowledge is 55.76% among primary school students in Shennongjia Forest District, and is affected by age, mother's educational level, average annual family income and living with parents.
5.Allele frequency polymorphism of Duffy, Kidd, Diego blood group system: a study in Dongxiang ethnic group in Gansu Province
Jun CUI ; Weiping ZHANG ; Xia HUI ; Xiaoping ZHANG ; Guoying LI ; Haiyan YANG
Chinese Journal of Blood Transfusion 2024;37(1):68-72
【Objective】 To investigate the allele frequencies polymorphic distribution of Duffy, Kidd and Diego blood group systems in Dongxiang ethnic group in Gansu Province. 【Methods】 Blood samples of 100 unrelated blood donors were randomly selected from Dongxiang ethnic group in Gansu from January to December 2017. Allelic typing of Duffy, Kidd and Diego blood groups was performed by fluorescence PCR. 【Results】 The allele frequencies of Duffy, Kidd, and Diego blood group systems of Dongxiang ethnic group were 0.835 for Fy*01, 0.165 for Fy*02, 0.570 for Jk*01, 0.430 for Jk*02, 0.020 for DI*01, 0.980 for DI*02, respectively. No Fy(a-b-), Jk(a-b-), Di(a+b-) rare phenotypes were found. The antigen incompatibility rates of Fya/Fyb, Jka/Jkb, Dia/Dib of Duffy, Kidd, and Diego blood group systems were 23.76%, 37.01% and 3.84%, respectively. 【Conclusion】 The allele frequencies distribution of Duffy, Kidd and Diego blood group systems in Dongxiang ethnic group in Gansu were polymorphic and has unique ethnic distribution characteristics.
6.Effect of storage temperature and time on coagulation factor after cryoprecipitated antihemophilic factor melting
Yuan WANG ; Guoying LIU ; Dawei KONG ; Jianbin LI ; Xinli JIN ; Yuhong ZHANG ; Wenchao GE ; Lin CHENG ; Jiaxuan LIU ; Yuzhen LIU
Chinese Journal of Blood Transfusion 2024;37(4):425-430
【Objective】 To study and compare the effects of different storage temperature and time on coagulation factor after cryoprecipitated antihemophilic factor(CAF) melting, and to provide reference for the establishment of industry standards. 【Methods】 From June 2021 to May 2023, a total of 96 bags of CAF were sampled in 4 bags per month, and timely detected in the same month. After the CAF was melted in a 37℃ water bath, the mild to moderate lipemic blood was labeled. Each bag of CAF and two 50 mL transfer bags were divided into two bags and two groups of 20 mL each using a sterile adapter. One group was placed in a 4℃ refrigerator and the other in a 22℃ water bath for 0 h, 4 h, 8 h, 12 h, 24 h and 48 h. Then 2 mL of aseptic sample was taken separately and put into the test tube, and 1mL of sample and 3 mL of buffer were added into the other test tube with the sampling gun and mixed on the machine for testing. The experimental data of 60 bags without mild to moderate lipemic blood cryoprecipitation and coagulation factor were randomly selected and statistically analyzed by SPSS21.0. 【Results】 After melting, CAF was stored for 0 h, 4 h, 8 h, 12 h, 24 h and 48 h to detect the average content and growth rate of coagulation factor in the two groups: 1) Storage at 4℃, factor Ⅷ content was 118.62, 111.57(-5.95%), 105.51(-11.05%), 103.30(-12.92%), 94.35(-20.46%) and 83.25(-29.82%) IU/ bag, respectively; Storage at 22℃, the factor Ⅷ content was 118.62, 112.69(-5.00%), 111.41(-6.08%), 109.01(-8.10%), 101.55(-14.39%) and 92.75(-21.81%) IU/ bag, and the storage results of the two groups were compared. At 24 h at 4℃ and 48 h at 22℃, the content of factor Ⅷ had significant statistical significance(P<0.01), and when stored at 22℃, the decay rate of factor Ⅷ was slower; 2) When stored at 4℃, the content of factor V was 41.19, 41.31(0.29%), 40.52(-1.64%), 40.27(-2.23%), 39.05(-5.19%) and 36.99(-10.21%) IU/ bag, respectively; Stored at 22℃, the factor V content was 41.19, 41.71(1.25%), 42.54(3.28%), 41.94(1.80%), 39.21(-4.80%) and 35.64(-13.48%) IU/ bag, respectively. Comparison of storage results between the two groups showed that the content of factor V was statistically significant(P<0.05) and significantly significant(P<0.01) at 4℃48 h and 22℃48 h, respectively, and the decay rate of factor V was faster when stored at 22℃; 3) When stored at 4℃, the Fbg content was 268.86, 268.17(-0.26%), 262.46(-2.38%), 270.50(0.61%), 267.52(-0.50%) and 261.92(-2.58%) mg/ bag, respectively; Stored at 22℃, the Fbg content was 268.86, 265.86(-1.12%), 264.12(-1.77%), 265.89(-1.11%), 266.04(-1.05%) and 261.04(-2.91%) mg/ bag, respectively. There was no statistical significance between the 2 groups and the original 0 h content in each time period(P>0.05). 【Conclusion】 After CAF melting, coagulation factor decreased with the extension of storage time, especially the decrease of factor Ⅷ, followed by factor V, while Fbg basically unchanged. Comparison between the two groups showed that, factor Ⅷ decay rate is slower, factor V decay rate is faster of storage at 22℃. CAF should be transfused as soon as possible after melting. If the delay is unavoidable, for the delay time less than 12 h, storage at 4℃ is recommended, fot the delay time more than 12 h and less than 24 h, storage at 22℃ is recommended.
7.Research Progress of Baitouweng Decoction and Its Modified Prescription in the Treatment of Ulcerative Colitis
Sihui QIN ; Yuan WANG ; Hui LI ; Jinge MA ; Guoying LIANG
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(10):186-190
Baitouweng Decoction is a classic prescription commonly used in the treatment of heat-toxin dysentery,with confirmed efficacy in the treatment of ulcerative colitis(UC).Clinical studies have found that Baitouweng Decoction,modified Baitouweng Decoction,combined with other Chinese materia medica or other therapies for UC can improve the clinical symptoms of patients,reduce the lesion activity score under colonoscopy,promote the healing of intestinal mucosal tissue,and also improve the overall efficiency of treatment,and reduce adverse reactions and recurrence rate.Pharmacological studies showed that Baitouweng Decoction may treat UC by regulating the expressions of related inflammatory factors and signaling pathways,regulating immune function,repairing intestinal mucosal barrier,and improving intestinal flora imbalance.This article systematically reviewed the clinical and basic research progress of Baitouweng Decoction in the treatment of UC,in order to provide a basis for relevant research.
8.Xie Jingri's Experience in Treating Cirrhotic Ascites Based on Collateral Disease Theory
Sihui QIN ; Hui LI ; Guoying LIANG ; Jingri XIE
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(12):176-179
This article introduced Professor Xie Jingri's clinical experience in explaining the etiology,pathogenesis and treatment of cirrhotic ascites based on the theory of collateral disease.Professor Xie Jingri believes that cirrhotic ascites is mostly the syndrome of deficiency in origin and excess in superficiality,and the mixture of deficiency and excess.Among them,the deficiency of collaterals is the root of the pathogenesis of cirrhotic ascites,and qi stagnation,water dampness,blood stasis and other pathogenic factors blocking collaterals are its superficiality.The treatment should be based on the basic principles of tonifying deficiency and dredging collaterals,promoting qi and dredging collaterals,removing dampness and dredging collaterals,and promoting blood circulation and dredging collaterals.One medical case about liver cirrhosis ascites of qi stagnation and collateral obstruction type was attached in this article.The treatment should be promoting qi and dredging collaterals,with confirmed clinical efficacy.
9.Association of T helper cells and cytokines with abdominal obesity in Hashimoto's thyroiditis
ZYNAT JAZYRA ; Suli LI ; Kaidi ZHANG ; Fuhui MA ; Guoying MA ; Yanying GUO
Basic & Clinical Medicine 2024;44(8):1120-1125
Objective To investigate the correlation between abdominal obesity and autoimmune thyroid disease in the view point of helper T cells and cytokines.Methods Clinical and laboratory data were collected from 108 pa-tients with Hashimoto's thyroiditis(HT)plus abdominal obesity and 122 patients of Hashimoto's thyroiditis without abdominal obesity who visited the People's Hospital of Xinjiang Uygur Autonomous Region and also from the control population.Abdominal circumference was measured,and patients in the HT patients were grouped according to whether they were abdominally obese or not.The thyroglobulin antibody(TgAb)and thyroid peroxidase antibody(TPOAb)were detected,and the ratio of helper T cells and related cytokines were detected by flow cytometry and enzyme-linked immunosorbent assay.Results The abdominal circumference of the TgAb-positive group was higher than that of the TgAb-negative group(P<0.05).Correlation analysis suggested that abdominal circumference was significantly and positively correlated with TgAb and IL-4 but negatively correlated with Th1.After correcting for gender and age,and abdominal obesity and IL-4 were risk factors for TgAb antibody positivity OR=3.080(95%CI:1.022-9.284)and OR=1.296(95%CI:1.022-9.284),both with P<0.05.Conclusions Abdominal obesity may be an influential factor in TgAb antibody positivity,with larger abdominal circumference having higher TgAb antibody titers,lower Th1 levels,and higher IL-4 levels.Visceral adiposity may exacerbate autoimmune dam-age of thyroid tissue by disruption of helper T cell pathway.
10.Association of copy number variation in X chromosome-linked PNPLA4 with heterotaxy and congenital heart disease
Han GAO ; Xianghui HUANG ; Weicheng CHEN ; Zhiyu FENG ; Zhengshan ZHAO ; Ping LI ; Chaozhong TAN ; Jinxin WANG ; Quannan ZHUANG ; Yuan GAO ; Shaojie MIN ; Qinyu YAO ; Maoxiang QIAN ; Xiaojing MA ; Feizhen WU ; Weili YAN ; Wei SHENG ; Guoying HUANG
Chinese Medical Journal 2024;137(15):1823-1834
Background::Heterotaxy (HTX) is a thoracoabdominal organ anomaly syndrome and commonly accompanied by congenital heart disease (CHD). The aim of this study was to analyze rare copy number variations (CNVs) in a HTX/CHD cohort and to examine the potential mechanisms contributing to HTX/CHD.Methods::Chromosome microarray analysis was used to identify rare CNVs in a cohort of 120 unrelated HTX/CHD patients, and available samples from parents were used to confirm the inheritance pattern. Potential candidate genes in CNVs region were prioritized via the DECIPHER database, and PNPLA4 was identified as the leading candidate gene. To validate, we generated PNPLA4-overexpressing human induced pluripotent stem cell lines as well as pnpla4-overexpressing zebrafish model, followed by a series of transcriptomic, biochemical and cellular analyses. Results::Seventeen rare CNVs were identified in 15 of the 120 HTX/CHD patients (12.5%). Xp22.31 duplication was one of the inherited CNVs identified in this HTX/CHD cohort, and PNPLA4 in the Xp22.31 was a candidate gene associated with HTX/CHD. PNPLA4 is expressed in the lateral plate mesoderm, which is known to be critical for left/right embryonic patterning as well as cardiomyocyte differentiation, and in the neural crest cell lineage. Through a series of in vivo and in vitro analyses at the molecular and cellular levels, we revealed that the biological function of PNPLA4 is importantly involved in the primary cilia formation and function via its regulation of energy metabolism and mitochondria-mediated ATP production. Conclusions::Our findings demonstrated a significant association between CNVs and HTX/CHD. Our data strongly suggested that an increased genetic dose of PNPLA4 due to Xp22.31 duplication is a disease-causing risk factor for HTX/CHD.


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