1.Role and mechanism of action of phytoestrogen biochanin A in improving liver fibrosis in ovariectomized mice
Chaorong TAN ; Xiaopiao LI ; Junyan RAN ; Ying XIONG ; Shanggao LIAO ; Jinjuan ZHANG ; Xun HE
Journal of Clinical Hepatology 2024;40(1):76-82
ObjectiveTo investigate the effect of phytoestrogen biochanin A (BCA) on liver fibrosis induced by CCl4 in female mice with bilateral oophorectomy (ovariectomized) and its mechanism. MethodsA total of 50 ovariectomized Kunming mice were selected and given intraperitoneal injection of CCl4 to establish a model of liver fibrosis, and then according to body weight, they were randomly divided into model group, positive control group, and low-, middle-, and high-dose BCA groups, with 10 mice in each group. In addition, 10 female mice in the same litter were given resection of a small amount of adipose tissue near both ovaries to establish the sham-operation group. The mice in the positive control group were given estradiol 2 mg/kg by gavage, and those in the low-, middle-, and high-dose BCA groups were given BCA by gavage at a dose of 25, 50, and 100 mg/kg, respectively, once a day for 7 consecutive weeks; the mice in the sham-operation group and the model group were given an equal volume of 0.5% sodium carboxymethyl cellulose solution by gavage. The mice were anesthetized and sacrificed after administration to collect samples. Liver index and uterus index were measured; HE staining and Masson staining were used to observe liver histopathological changes; the biochemical analysis was used to measure the activity of aspartate aminotransferase (AST) and alanine aminotransferase (ALT); ELISA was used to measure the levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in liver tissue, and Western blot was used to measure the relative protein expression levels of collagen Ⅰ, transforming growth factor-beta 1 (TGF-β1), alpha-smooth muscle actin (α-SMA), estrogen receptor beta (ERβ), and p-NF-κBp65/NF-κBp65 in liver tissue. The t-test was used for comparison of continuous data between two groups; a one-way analysis of various was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups and further comparison between two groups. ResultsCompared with the sham-operated group, the model group had a significant increase in liver index and a significant reduction in uterus index, as well as significant increases in the activities of serum AST and ALT, the levels of IL-6 and TNF-α in liver tissue, and the protein expression levels of collagen Ⅰ, TGF-β1, α-SMA, and p-NF-κBp65/NF-κBp65 in liver tissue (all P<0.05), with no significant change in the expression of ERβ in liver tissue (P>0.05), and the model group showed significant fibrosis lesions in the liver, such as hepatocyte edema, steatosis, and necrosis with inflammatory cell infiltration and hyperplasia, deposition, and staggered distribution of collagen fibers. Compared with the model group, the low-, middle-, and high-dose BCA groups had significant reductions in liver index, the activities of serum AST and ALT, the levels of IL-6 and TNF-α, and the protein expression levels of collagen Ⅰ, TGF-β1, α-SMA, and p-NF-κBp65/NF-κBp65 in liver tissue (all P<0.05), with no significant change in uterine index (P>0.05), as well as a significant increase in the protein expression level of ERβ in liver tissue (P<0.05) and varying degrees of improvement in liver fibrosis lesions. ConclusionBCA can effectively improve CCL4-induced liver fibrosis in ovariectomized female mice, possibly by upregulating ERβ to inhibit the NF-κB signaling pathway and then alleviating inflammatory response.
2.Discussion on the pathogenesis and treatment of non-alcoholic fatty liver disease in postmenopausal women based on the theory of "deficient qi and stagnation"
Mingyao XU ; Wenyuan SU ; Jing WU ; Jiawen HE ; Haowei ZHANG ; Jinjuan LI ; Jia YUE ; Yan ZHANG
Journal of Beijing University of Traditional Chinese Medicine 2024;47(12):1745-1752
Non-alcoholic fatty liver disease is a common chronic liver disease in clinical practice. In recent years, with increasing social attention to the health of women and the elderly, the prevention and treatment of non-alcoholic fatty liver disease after menopause has increasingly become a research hotspot in metabolic diseases. This study explores the pathogenesis and treatment method of non-alcoholic fatty liver disease in postmenopausal women based on the theory of "deficient qi and stagnation" and combined with the physiological and pathological characteristics of postmenopausal women and the Western medicine understanding of non-alcoholic fatty liver disease. We believe that the pathogenesis of non-alcoholic fatty liver disease in postmenopausal women is rooted in the "deficient qi" caused by depletion of liver and kidney essence and blood. The imbalance between the physical and functional aspects of the liver due to this "deficient qi" is the primary factor, while the "stagnation" of phlegm and blood stasis is the manifestation. Furthermore, the "deficient qi" and "stagnation" reinforce each other, with the deficiency leading to stagnation and stagnation exacerbating the deficiency, thus accelerating the progression of the disease. The treatment approach should be one that combines nourishing deficiency and resolving stagnation, addressing both root cause and maifestations. Given the female characteristic of "the liver as the innate organ" and the post-menopausal physiological state of "gradual decline of kidney essence", it is important to focus on nourishing the liver and kidneys, nurturing the liver′s physical body while maintaining its function, and also promoting the circulation of qi, resolving phlegm, and invigorating blood circulation to remove blood stasis. This approach aims to reduce the accumulation of lipids in the liver, offering a new perspective and approach for the treatment of non-alcoholic fatty liver disease in post-menopausal women with traditional Chinese medicine.
3.Lycium barbarum polysaccharide inhibits starvation-induced apoptosis of Schwann cells by enhancing autophagy
Yumei LIU ; Heying WANG ; Caihong WEI ; Jinjuan WANG ; Junping LI ; Hua JIA
Chinese Journal of Neuroanatomy 2024;40(1):16-24
Objective:To investigate the inhibitory effect of lycium barbarum polysaccharide(LBP)on apoptosis of Schwann cells(SCs)and its related mechanisms.Methods:The autophagy model was prepared by starvation treatment of RSC96 cells for 12 h,and the expressions of autophagy related proteins LC3 and p62 were detected by Western Blot.Cell Counting Kit-8(CCK-8)kits were used to detect the optimal concentration of LBP.RSC96 cells were randomly divided into Control group,Starvation group and Starvation+LBP group.The expressions of autophagy associated pro-teins(LC3,p62)and myelin associated proteins(p75NTR,PMP22,S100β)were detected by Western Blot or immu-nofluorescence staining.Annexin V/PI fluorescence staining was used to detect apoptosis of the cells.The cell cycle was analyzed by flow cytometry.Western Blot analysis of phosphorylation levels of pathway proteins Erk1/2 and Akt.Results:CCK8 results showed that the viability of damaged RSC96 cells was the best when LBP was 300 μg/ml.Com-pared with Control group,LC3-Ⅱ/LC3-I levels in Starvation group were significantly increased(P<0.05).Compared with Starvation group,the proportion of apoptotic and necrotic cells in Starvation+LBP group was significantly de-creased,and the proportion of cells in S and G2/M stages was increased.The expression levels of LC3-Ⅱ,p75NTR,PMP22 and S100β were increased,while the expression levels of autophagy substrate protein p62 were decreased.In-creased expression of pathway protein p-Erk1/2(P<0.05),while the expression of p-Akt protein decreased slightly.Conclusion:LBP can inhibit the apoptosis of SCs and promote the expression of myelin-related proteins by enhancing autophagy,which is related to the activation of Erk1/2 and/or the inhibition of Akt.
4.Comprehensive protein kinase inhibition analysis reveals the molecu-lar mechanism of KG-1 proliferation
Yu DUAN ; Ningxin XU ; Qiong CAO ; Kai YANG ; Jinjuan WANG ; Sijin LIU ; Fengfeng JIA ; Jianbing LIU ; Li LI
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(6):621-628
AIM:To investigate the molecular mechanisms of KG-1 cell proliferation by profiling its responses to various protein kinase inhibitors.METHODS:CCK-8 assay,real time quantitative PCR(qRT-PCR)and Western-blot were used to detect the effect of various protein kinase inhibitors on KG-1 cell proliferation,the expression levels of mRNA and phosphorylation level of signaling pro-teins in the FGFR1 downstream pathways.RE-SULTS:NVP-BGJ398 and PD173074 effectively in-hibited the proliferation of KG-1 cells,indicative of a crucial role of FGFR downstream signaling.After treatment with FGFR inhibitors,the levels of p-FG-FR1OP2-FGFR1 and p-STAT5 decreased significantly(P<0.001),p-AKT decreased slightly(P<0.05),with-out affecting the p-ERK level(P>0.05).CONCLU-SION:FGFR1OP2-FGFR1 mainly acts on the down-stream STAT5 signaling pathway to promote cell proliferation.Comprehensive protein kinase inhibi-tion analysis is a reliable and direct approach to identify functional drivers of cancer cell prolifera-tion.
5.A pedigree of pseudohypoaldosteronism type Ⅱ and review of literature
Lixin DING ; Ping ZHANG ; Hui WANG ; Li FENG ; Xiaohua GUAN ; Qimei YANG ; Li ZHU ; Wenlong XU ; Xinping ZHANG ; Liyuan LI ; Jinjuan CAO
Chinese Journal of Endocrinology and Metabolism 2024;40(6):521-524
The clinical data, laboratory test, and gene mutations were collected from a family with pseudohypoaldosteronism type II(PHA2). The proband, aged 1 year and 7 months, presented with hyperkalemia(6.69 mmol/L; reference range 3.5-5.3 mmol/L), blood pressure of 110/68 mmHg(normal<106/61 mmHg, 1 mmHg=0.133 kPa), blood chloride of 111.5 mmol/L(reference 99-110 mmol/L), blood HCO 3- of 17.1 mmol/L(reference 22-29 mmol/L), estimated glomerular filtration rate of 128.5 mL·min -1·(1.73 m 2) -1[>90 mL·min -1·(1.73 m 2) -1], and blood renin concentration of 0.30 μIU/mL(reference 4.2-45.6 μIU/mL). The mother and maternal grandfather also exhibited normal renal function with hyperkalemia, hypertension, hyperchloremia, metabolic acidosis, and low renin. Genetic testing revealed a heterozygous missense mutation(c.1685A>G, p. E562G) in exon 7 of the no-lysine kinase 4(WNK4) gene. Treatment with hydrochlorothiazide was effective. Literature review comparing this E562G pedigree with other WNK4 variants suggested clinical heterogeneity of WNK4 mutations. For unexplained hyperkalemia, especially with concurrent hypertension, PHA2 should be considered early for genetic screening to prevent misdiagnosis or delayed diagnosis.
6.Protective effects of twin drug ST-11 against PC12 cell injury induced by oxygen-glucose deprivation/reperfusion and its mechanism
Jinlan WEN ; Lina CHEN ; Shanhui ZHANG ; Jun LU ; Shuxia WU ; Jinjuan ZHANG ; Yongxi DONG ; Li DONG
China Pharmacy 2023;34(6):660-665
OBJECTIVE To study the protective effects of ligustrazine-scutellarein twin drug ST-11 on rat adrenal medullary pheochromocytoma PC12 cell injury induced by oxygen-glucose deprivation/reperfusion (OGD/R) and its mechanism. METHODS PC12 cells were divided into blank group, model group, nimodipine group (positive control, 5 μmol/L) and different concentration groups of ST-11 (5, 10, 20 μmol/L). After 24 hours of pre-administration intervention, all the other groups except the blank group were cultured in glucose-free DMEM culture medium containing 10 mmol/L Na2S2O4 for 4 hours with glucose deficiency and hypoxia. After 4 hours of glucose and oxygen re-introduction, the survival rate of cells in each group, the contents of lactate dehydrogenase (LDH), catalase (CAT), glutathione (GSH), malondialdehyde (MDA) and superoxide dismutase (SOD) in cell supernatant, apoptosis rate, the levels of reactive oxygen species (ROS) and mitochondrial membrane potential (MMP), the protein expressions of B-cell lymphoma 2 related X protein (Bax), B-cell lymphoma 2 (Bcl-2) and caspase-3 were all detected in each group. RESULTS Compared with blank group, the cell survival rate, the contents of CAT, GSH and SOD in cell supernatant, MMP level, relative expression of Bcl-2 and Bcl-2/Bax ratio in model group decreased significantly (P<0.05), while the contents of LDH and MDA, ROS level, apoptosis rate, relative expressions of Bax and caspase-3 were significantly increased (P<0.05). Compared with model group, above indexes of ST-11 groups (except for the protein expression of caspase-3 in 5 μmol/L ST-11 group) were reversed signifi-cantly (P<0.05). CONCLUSIONS ST-11 has a certain protec-tive effect on OGD/R-injured PC12 cells, and its effects may be related to reduction of oxidative stress and inhibition of cell apoptosis.
7.Preliminary Study on Improvement Effect of Tiarella polyphylla Ethanol Extract on CCl 4-induced Hepatic Fibrosis in Mice and Its Mechanism
Fujing HUANG ; Jinjuan ZHANG ; Li DONG ; Diao LI ; Chunlei ZHANG ; Shanggao LIAO ; Xun HE
China Pharmacy 2021;32(14):1685-1691
OBJECTIVE:To investigate the improvement effect of Tiarella polyphylla ethanol extract (TPME)on CCl 4-induced hepatic fibrosis in mice ,and to explore its possible mechanism preliminarily. METHODS :Totally 60 male Kunming mice were randomly divided into normal group ,model group ,positive control group (colchicine 0.1 mg/kg),TPME low-dose ,medium-dose and high-dose groups (250,500,1 000 mg/kg)according to body weight ,with 10 mice in each group. Except for normal group , other groups were given 20% CCl4 olive oil solution intraperitoneally to induce hepatic fibrosis ,twice a week ,for consecutive 8 weeks. From the fifth week after modeling ,administration groups were given relevant medicine intragastrically ,normal group and model group were given constant volume of normal saline intragastrically ,once a day ,for consecutive 4 weeks. Twelve hours after last administration ,the liver weight of mice in each group was measured and the liver index was calculated. The serum contents of ALT,AST,SOD,MDA,PC-Ⅲ,C-Ⅳ,LN,TNF-α and IL- 6 were determined. Western blot assay was used to detect the protein expression of α-SMA,TGF-β1 and Smad 3 in liver tissue. HE and Masson staining were used to observe the pathological changes of hepatic tissue. RESULTS :Compared with normal group ,the liver index ,the activities of ALT and AST and the contents of MDA , LN,PC- Ⅲ ,C- Ⅳ ,LN,TNF-α and IL- 6 in serum were increased significantly , while the activity of SOD was 6011) decreased significantly in model group (P<0.01);the protein expression of α-SMA,TGF-β1 and Smad 3 in liver tissues were hfjsznd8@126.com increased significantly (P<0.01). Obvious fibrosis lesions was observed in liver tissue. Compared with model group ,the live indexes ,the activities of ALT and AST ,the contents of MDA,PC-Ⅲ,C-Ⅳ,LN,TNF-α and IL-6 in serum were decreased significantly in positive control group and TPME groups , while the activities of SOD were increased significantly (P<0.05 or P<0.01). The protein expression of α-SMA,TGF-β1 and Smad3 in liver tissue were decreased significantly (P<0.05 or P<0.01),and liver fibrosis was improved to different extent. Compared with TPME low-dose group ,the contents of PC- Ⅲ,LN and IL- 6 in serum ,protein expression of TGF-β1 and Smad 3 in liver tissue were decreased significantly in TPME high-dose group (P<0.05). CONCLUSIONS :TPME can improve hepatic fibrosis induced by CCl 4 in mice ,the mechanism of which may be associated with the inhibition of collagen synthesis and oxidative stress,the reduction of inflammatory factors ,and the down-regulation of the expression α-SMA and relative proteins of TGF-β1/ Smad signal pathway.
8.Unqualified rate of anti-HIV detection in the laboratories from blood banks in Beijing-Tianjin-Hebei region
Wei ZHEN ; Hongwei GE ; Rui WANG ; Tong PAN ; Wei HAN ; Peng WANG ; Li YANG ; Shaoqiu SUN ; Xiao CAO ; Liye CUI ; Chao WEI ; Guijun YU ; Yunpeng XU ; Jinjuan FANG ; Caixia LIU ; Xuegang WANG ; Zhijun ZHEN ; Xiaojie LIU ; Wengong DU ; Lunan WANG ; Jiang LIU ; Hongjie WANG
Chinese Journal of Blood Transfusion 2021;34(4):371-376
【Objective】 To investigate the unqualified rate of anti-HIV detection of blood screening laboratories in Beijing-Tianjin-Hebei region, and explore the differences in anti-HIV detection ability and influencing factors in each laboratory. 【Methods】 Through filling questionnaires via e-mail, the anti-HIV ELISA unqualified rate and confirmed (WB) positive results (data) from January to December 2018 from 15 blood screening laboratories in Beijing-Tianjin-Hebei region were collected. Our laboratory was responsible for data collection and confirmation, and statistics software SPSS22.0 was used for analysis. 【Results】 1) There was a statistically significant difference among the unqualified rate of anti-HIV ELISA(6.77‱~35.71‱) and confirmed positive rate(0.60‱~3.56‱) in 15 blood screening laboratories in Beijing-Tianjin-Hebei region (P<0.05); 2) There were significant differencse among the ELISA unqualified rate and the confirmed positive rate of 8 reagents for anti-HIV detection(P<0.01), and the sensitivity of the 4th generation detection reagent and the imported reagent was higher than that of the 3rd generation reagent and the domestic reagent. The anti-HIV ELISA unqualified rate of R5 was the highest (19.08‱). 3)There were significant differences in the anti-HIV ELISA unqualified rate of R1, R2, R3, R5 and R7 reagents among different blood station laboratories(P<0.05), and there were no significant differences in the anti-HIV ELISA unqualified rate of R4, R6 and R8 reagents among different blood station laboratories(P>0.05). 4)The unqualified rate of anti-HIV ELISA of laboratories using different regents showed significant differences(P<0.05), except H, J, M. The unqualified rate of imported reagent was significantly higher than that of domestic reagents of laboratories using imported and domestic reagents combinations(P<0.05), except O. 62.5% (5/8) laboratories using domestic 3rd and 4th generation reagent combination showed significant differences in the unqualified rates among different reagents(P<0.05); 5) The positive rate of single-reagent(62.02%~95.45%)in 15 blood screening laboratories showed significant difference(P<0.001), and A was the lowest (62.02%). 【Conclusion】 The anti-HIV detection ability among 15 blood screening laboratories in Beijing-Tianjin-Hebei region is quite different. The application of different reagents is the main factor for the difference, and other factors such as personnel, instruments and test strategies also has a great impact on the detection of anti-HIV. It is still necessary to promote the process of homogenization of blood testing quality among blood screening laboratories in Beijing-Tianjin-Hebei region.
9.Inhibitory Effects of Protopine on the Proliferation of Human Hepatic Stellate Cells HSC-LX 2 and Its Mechanism Study
Xinyu WU ; Jing LI ; Jinjuan ZHANG ; Shanggao LIAO ; Qing MEI ; Yayun WU ; Xiaolan XI
China Pharmacy 2019;30(23):3210-3215
OBJECTIVE: To investigate inhibitory effects of protopine on the proliferation of human hepatic stellate cells HSC-LX2 and to explore its mechanism preliminarily. METHODS: MTT assay was used to detect the effects of 25, 50, 100, 200, 400 and 500 μmol/L protopine on the proliferation of HSC-LX2 cells. The inhibitory effect of cell proliferation was calculated. HSC-LX2 cells were divided into control group (1640 medium containing 5% fetal bovine serum), protopine low-concentration, medium-concentration and high-concentration groups (100, 200, 400 μmol/L). After treated for 24 h. The apoptotic rate of the cells was detected by flow cytometry. RT-PCR was used to determine the mRNA expression of α-SMA, Collagen Ⅰ, Collagen Ⅲ, MMP-2 and TIMP-1 in cells. The protein expressions of α-SMA, Collagen Ⅰ, Collagen Ⅲ and MMP-2 were detected by Western blot. RESULTS: The inhibitory rates of 25, 50, 100, 200, 400 and 500 μmol/L protopine on proliferation HSC-LX2 cells were 0, 6.9%, 18.7%, 34.2%, 48.9%, 53.9%, respectively. Compared with control group, mRNA expression of Collagen Ⅰ, TIMP-1 and protein expression of α-SMA were decreased significantly in protopine low-concentration, medium-concentration and high-concentration groups, while protein expression of MMP- 2 was increased significantly, with statistical significance (P<0.05 or P<0.01). Apoptotic rate of HSC-LX2 cells and mRNA expression of MMP-2 were increased significantly in protopine medium-concentration and high-concentration groups, mRNA expression of α-SMA and Collagen Ⅲ, protein expression of Collagen Ⅰ and Collagen Ⅲ were decreased significantly, with statistical significance (P<0.05 or P<0.01). CONCLUSIONS: Protopine can induce the apoptosis of HSC-LX2 cells and inhibit their cell proliferation, and reduce the expression of a-SMA, Collagen Ⅰ, Collagen Ⅲ and TIMP-1, and increase the expression of MMP-2.
10.An investigation on iodine nutrition and thyroid stimulating hormone level of patients with thyroid nodules in different regions of Qinghai Province
Na MI ; Zheng LI ; Xianjin CHONG ; Haiqi XU ; Yonggui LI ; Ye WANG ; Qinfang ZHU ; Jinjuan LI ; Lixin YANG ; Guotong LI ; Zhixiao JIANG
Chinese Journal of Endemiology 2018;37(7):565-567
Objective To investigate the iodine nutritional and thyroid stimulating hormone level of patients with thyroid nodules in different regions of Qinghai Province and analyze the characteristics of changes in different regions.Methods In 2014-2016,thyroid nodules in 9 regions of Qinghai Province (Tibetan areas:Xiewu,Nangqian,Jiegu,Guoluo;non-Tibetan areas:Xining,Huzhu,Menyuan,Minhe,and Ledu) were selected and serum thyroid stimulating hormone (TSH) and urine iodine were measured.Results A total of 553 thyroid nodules,the median urinary iodine (MUIC) was 160.8 μg/L and the median TSH was 2.97 mU/L.The iodine nutritional status was at an appropriate level.Among them,MUIC (206.8 μg/L) in thyroid nodules in the Menyuan area was slightly higher than the appropriate amount,there was a significant difference in MUIC among different region (x2 =47.747,P < 0.05);of TSH in thyroid nudules in the 9 regions,the differences were statistically significant (x2 =34.832,P < 0.05).Non-Tibetan areas were compared with Tibetan areas,there was a significant difference in MUIC (155.6,185.6 μg/L),TSH (2.68,3.45 mU/L,Z =-3.677,-5.410,P < 0.05);Among them,the differences was statistically significant between MUIC (152.8,187.7 μg/L) of women with thyroid nodules (Z =-3.504,P < 0.05);there was a statistically significant difference in TSH levels among men (2.58,3.46 mU/L) and women (2.80,3.44 mU/L) with thyroid nodules (Z =-3.613,-4.040,P < 0.05);there were no significant differences in MUIC levels among thyroid nodules of each age groups (P > 0.05);of the TSH level in 30-and 50-< 65 years groups (2.63,3.17;2.25,3.58 mU/L),the differences were statistically significant (Z =-2.892,-3.233,P < 0.05),and other groups were no significant differences (P > 0.05).Conclusion The iodine nutrition of patients with thyroid nodules in these regions of Qinghai Province is generally at an appropriate level,the MUIC and TSH levels in Tibetan areas were lower than those in non-Tibetan areas,and iodine nutrition status and TSH levels should be monitored for key populations.


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