1.Effect of Huatan Qushi Huoxue prescription on macrophage efferocytosis mediated by a disintegrin and metalloproteinase 17 and triggering receptor expressed on myeloid cells 2 in rats with metabolic dysfunction-associated steatohepatitis
Lihui ZHANG ; Sutong LIU ; Qing ZHAO ; Shanzheng LI ; Minghao LIU ; Wenxia ZHAO
Journal of Clinical Hepatology 2026;42(2):345-355
ObjectiveTo investigate the therapeutic effect and mechanism of Huatan Qushi Huoxue prescription on rats with metabolic dysfunction-associated steatohepatitis (MASH). MethodsA total of 60 specific pathogen-free Sprague-Dawley rats were randomly divided into blank control group, model A group, model B group, Western medicine group (polyene phosphatidylcholine, 143.64 mg/kg), high-dose Chinese medicine group (Huatan Qushi Huoxue prescription, 20.16 g/kg), and middle-dose Chinese medicine group (Huatan Qushi Huoxue prescription, 10.08 g/kg). All rats except those in the blank control group were given high-fat diet. Samples were collected from the model A group at week 8, and since week 12, the other groups were given the corresponding drug once a day for 8 consecutive weeks, with samples collected at week 20. Body weight, liver wet weight, and liver index were measured for all rats; the microplate method was used to measure the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and free fatty acids (FFA); ELISA was used to measure the serum levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and soluble triggering receptor expressed on myeloid cells 2 (sTREM2); HE staining and oil red O staining were performed to observe liver histopathological changes; immunofluorescence assay was used to measure CD68+TREM2+ cells in liver tissue and calculate the phagocytosis rate of macrophages; quantitative real-time PCR was used to measure the mRNA expression levels of sphingosine 1-phosphate (S1P), sphingosine 1-phosphate receptor 1 (S1PR1), a disintegrin and metalloproteinase 17 (ADAM17), and triggering receptor expressed on myeloid cells 2 (TREM2) in liver tissue, and immunohistochemistry was used to measure the protein expression levels of S1P, S1PR1, ADAM17, and TREM2 in liver tissue. A one-way analysis of variance was used for comparison of normally distributed continuous data with homogeneity of variance between groups, and the least significant difference t-test was used for further comparison between two groups; the Welch’s test was used for comparison of normally distributed continuous data with heterogeneity of variance between groups, and the Tamhane’s 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 groups, and the Dunn’s test was used for further comparison between two groups. ResultsCompared with the blank control group, the model A group and the model B group had significant increases in body weight and liver wet weight, and the model B group had a significant increase in liver index (all P<0.05). HE staining showed diffuse macrovesicular steatosis of liver tissue in the model A group and a large number of hepatocytes with ballooning degeneration in liver tissue in the model group B, with the presence of mixed inflammatory cell infiltration and mild perisinusoidal fibrosis in the lobules and the portal area. Compared with the blank control group, the model A group and the model B group had significant increases in NAS score and oil red O-positive area (all P<0.05), and the model B group had significant increases in these two indicators than the model A group (both P<0.05). Compared with the blank control group, the model A group and the model B group had significant increases in the serum levels of TC, TG, LDL-C, FFA, IL-1β, IL-6, and sTREM2 and a significant reduction in the serum level of HDL-C, and the model B group had significant increases in the serum levels of ALT, AST, and TNF-α (all P<0.05); compared with the model A group, the model B group had significant increases in the serum levels of ALT, AST, TC, TG, FFA, TNF-α, IL-1β, IL-6, and sTREM2 and a significant reduction in the serum level of HDL-C (all P<0.05). Immunofluorescence assay showed that compared with the blank control group, the model A group had a significant increase in the phagocytosis rate of macrophages (P<0.05), while the model B group had a significantly lower phagocytosis rate of macrophages than the model A group (P<0.05). Quantitative real-time PCR showed that compared with the blank control group, the model A group and the model B group had a significant increase in the mRNA expression level of TREM2, and the model B group had significant increases in the mRNA expression levels of S1P and S1PR1 (both P<0.05); moreover, compared with the model A group, the model B group had significant increases in the mRNA expression levels of S1PR1 and TREM2 (both P<0.05). Immunohistochemistry showed that compared with the blank control group, the model A group and the model B group had significant increases in the protein expression levels of S1P, S1PR1, and ADAM17, and the model A group had a significant increase in the protein expression level of TREM2 (all P<0.05); compared with the model A group, the model B group had significant increases in the protein expression levels of S1P, S1PR1, and ADAM17 and a significant reduction in the protein expression level of TREM2 (all P<0.05). Compared with the model B group, each medication group had significant reductions in body weight, liver wet weight, and liver index (all P<0.05); each medication group had significant improvements in hepatic steatosis and inflammatory damage, with significant reductions in NAS score and oil red O-positive area (all P<0.05); each medication group had significant reductions in the serum levels of ALT, AST, TC, TG, FFA, IL-1β, and IL-6 (all P<0.05) and a significant increase in the serum level of HDL-C (P<0.05), and the high-dose Chinese medicine group had a significant reduction in the serum level of TNF-α (P<0.05); each medication group had a significant increase in the phagocytosis rate of macrophages (all P<0.05); the high- and middle-dose Chinese medicine groups had a significant reduction in the protein expression level of ADAM17, and the high-dose Chinese medicine group had a significant increase in the protein expression level of TREM2 (all P<0.05). ConclusionHuatan Qushi Huoxue prescription improves lipid metabolism and inflammation in the liver of MASH rats by regulating hepatic macrophage phagocytosis.
2.Exploration on the Mechanism of Bushen Zhupai Decoction against Ovarian Fibrosis in Polycystic Ovary Syndrome Based on IL-17/TRAF6/NF-κB Signaling Pathway
He LI ; Wenjuan LIN ; Shuyuan ZHANG ; Shanzheng LI ; Cancan HUANG ; Rongrong YUAN ; Wenwen WAN ; Shouyue RUI ; Haiyan MAO ; Xiaohua ZHANG ; Quansheng WU
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(5):106-113
Objective To explore the mechanism of Bushen Zhupai Decoction in treating ovarian fibrosis in polycystic ovary syndrome(PCOS)model rats based on IL-17/TRAF6/NF-κB signaling pathway.Methods Totally 60 SPF grade female SD rats were randomly divided into blank group,model group,Yousiyue group and TCM low-,medium-and high-dosage groups,with 10 rats in each group.The PCOS model was replicated by gavage of 1 mg/kg letrozole for 21 days.The Yousiyue group was given a 10 mg/kg solution of drospirenone ethinylestradiol tablets by gavage,TCM low-,medium-and high-dosage groups were given 8.82,17.64 and 35.28 g/kg of Bushen Zhupai Decoction by gavage,once a day for 28 consecutive days.HE and Masson staining were used to observe the morphology of ovarian tissue and ovarian fibrosis,the contents of serum IL-17,IL-18 and IL-1β were detected by ELISA,the mRNA expression of IL-17,TRAF6,NF-κB p65 and TGF-β1 in ovarian tissue were determined by RT-qPCR,the protein expression of TRAF6,NF-κB p65,TGF-β1,α-SMA,E-cadherin and the positive expression of IL-17RA,TRAF6,NF-κB p65,TGF-β1 and α-SMA in ovarian tissue were detected by Western blot and immunohistochemical staining respectively.Results Compared with the blank group,the body mass,ovarian mass and ovarian index significantly increased of rats in model group,with follicular cystic dilation,thinning of granulosa cell layer,and significant increase in fibrosis positive range(P<0.01),the serum contents of IL-17,IL-18 and IL-1β significantly increased(P<0.01),the expressions of IL-17,TRAF6,NF-κB p65,TGF-β1 mRNA and IL-17RA,TRAF6,NF-κB p65,TGF-β1 and α-SMA proteins significantly increased(P<0.01,P<0.05),the expression of E-cadherin protein significantly decreased(P<0.01).Compared with the model group,the ovarian index of rats in Yousiyue group and TCM medium-and high-dosage groups significantly decreased(P<0.01,P<0.05),the rats in TCM high-dosage group showed dominant follicles,increased thickness of granulosa cell layer,and significantly reduced fibrosis positive range(P<0.01),the serum contents of IL-17,IL-18 and IL-1β significantly decreased in Yousiyue group and TCM low-,medium-and high-dosage groups(P<0.01),the mRNA expressions of IL-17,TRAF6,NF-κB p65,TGF-β1,and the protein expressions of IL-17RA,TRAF6,NF-κB p65,TGF-β1 and α-SMA in ovarian tissue significantly decreaed in TCM high-dosage group(P<0.01,P<0.05),the expression of E-cadherin protein significantly increased(P<0.01).Conclusion Bushen Zhupai Decoction can decrease the body mass,improve the ovarian index,decrease the expressions of inflammatory factors and improve the status of ovarian fibrosis in PCOS rats.The mechanism may be related to the regulation of IL-17/TRAF6/NF-κB signaling pathway.
3.Study on the protective efect and mechanism of paeoniflorin on palmitic acid-induced HepG2 cells
Tong LIU ; Shanzheng LI ; Cheng ZHOU ; Sutong LIU ; Lihui ZHANG ; Wenxia ZHAO
Journal of Clinical Hepatology 2025;41(3):499-505
ObjectiveTo investigate the role and mechanism of action of paeoniflorin (PF) in protecting HepG2 cells induced by palmitic acid (PA). MethodsHepG2 cells were stimulated with PA at a concentration of 250 μmol/L to establish a NAFLD model. Compound C at a concentration of 10 μmol/L was used as an inhibitor, and PF at a concentration of 25 μmol/L was used for intervention. The experiment was divided into normal group (CON group) treated with complete culture medium, model group (MOD group) treated with PA, PF treatment group (MOD+PF group) treated with PA and PF, model+inhibitor group (MOD+COM group) treated with PA and Compound C, and model+inhibitor+PF group (MOD+COM+PF group) treated with PA, Compound C, and PF. Kits were used to measure lipid deposition indicators, liver function parameters, oxidative stress indicators, and inflammation indicators; oil red O staining was used to observe lipid deposition; Western Blot was used to measure the protein expression levels of AMPK, SIRT1, PGC-1α, mTOR, Beclin-1, LC3, and P62 in cells. The one-way analysis of variance was used for comparison of quantitative data between groups, while the Tukey’s test was used for comparison between two groups. ResultsCompared with the MOD group, PF improved the levels of TC and TG (P<0.05), reduced the levels of ALT, AST, CRP, TNF-α, IL-1β, and IL-6 (P<0.05), increased the activity of SOD and CAT and the level of GSH, and reduced the level of MDA in cells (all P<0.05). Oil red O staining showed that PF alleviated lipid deposition in cells. Western blot results showed that compared with the MOD group, PF increased the protein expression levels of p-AMPK, SIRT1, PGC-1α, LC3Ⅱ/LC3Ⅰ, and Beclin-1 and reduced the protein expression levels of p-mTOR and P62 (all P<0.05). ConclusionPF can inhibit PA-induced oxidative stress and inflammatory response in HepG2 cells, improve lipid deposition, and promote autophagy via the AMPK/SIRT1/PGC-1α/mTOR signaling pathway.
4.Construction of an evaluation scale for non-alcoholic fatty liver disease with internal retention of dampness and turbidity based on the method of combining disease and syndrome
Cheng ZHOU ; Jinqiu YANG ; Tong LIU ; Shanzheng LI ; Tong LIU ; Wenxia ZHAO
Journal of Clinical Hepatology 2025;41(4):650-660
ObjectiveTo develop a scale for evaluating the syndrome of internal retention of dampness and turbidity in nonalcoholic fatty liver disease (NAFLD) that combines disease and syndrome and has the characteristics of traditional Chinese medicine (TCM). MethodsAn item pool was established for evaluating the syndrome of internal retention of dampness and turbidity in nonalcoholic fatty liver disease (NAFLD) with reference to the guideline for developing international scales. A clinical survey was conducted among the outpatients and inpatients who were diagnosed with NAFLD and had the syndrome of internal retention of dampness and turbidity in Department of Hepatology and Spleen-Stomach, The First Affiliated Hospital of Henan University of Chinese Medicine, from June to August, 2023, and the items were screened based on the classical test theory and the item response theory. An expert questionnaire was developed, and expert discussions were conducted using the Delphi method to identify the items for evaluating the syndrome of internal retention of dampness and turbidity in NAFLD. Finally, the scale was given scientific scores. ResultsA preliminary item pool was established, with 16 primary items and 22 secondary items, and it was divided into the two dimensions of disease and syndrome type. Clinical pre-survey suggested to retain 9 primary items and 14 secondary items, while the Delphi expert questionnaire recommended to retain 11 primary items and 15 secondary items, and tongue manifestation and pulse manifestation were no longer used for assessing the severity of syndrome. After hierarchical analysis and scientific assignment of scores, the scale for evaluating the syndrome of internal retention of dampness and turbidity in NAFLD had a total score of 123 points and 9 important items, i.e., discomfort in the hypochondrium, abdominal fullness and distension, obesity, heaviness of the head and body, loose stool, anorexia, coughing up phlegm, nausea with a tendency to vomit, and lethargy. ConclusionA preliminary scale is established for evaluating the syndrome of internal retention of dampness and turbidity in NAFLD, which fills the gap in this research field and provides a basis for further clinical application.
5.Exploration on the Mechanism of Bushen Zhupai Decoction against Ovarian Fibrosis in Polycystic Ovary Syndrome Based on IL-17/TRAF6/NF-κB Signaling Pathway
He LI ; Wenjuan LIN ; Shuyuan ZHANG ; Shanzheng LI ; Cancan HUANG ; Rongrong YUAN ; Wenwen WAN ; Shouyue RUI ; Haiyan MAO ; Xiaohua ZHANG ; Quansheng WU
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(5):106-113
Objective To explore the mechanism of Bushen Zhupai Decoction in treating ovarian fibrosis in polycystic ovary syndrome(PCOS)model rats based on IL-17/TRAF6/NF-κB signaling pathway.Methods Totally 60 SPF grade female SD rats were randomly divided into blank group,model group,Yousiyue group and TCM low-,medium-and high-dosage groups,with 10 rats in each group.The PCOS model was replicated by gavage of 1 mg/kg letrozole for 21 days.The Yousiyue group was given a 10 mg/kg solution of drospirenone ethinylestradiol tablets by gavage,TCM low-,medium-and high-dosage groups were given 8.82,17.64 and 35.28 g/kg of Bushen Zhupai Decoction by gavage,once a day for 28 consecutive days.HE and Masson staining were used to observe the morphology of ovarian tissue and ovarian fibrosis,the contents of serum IL-17,IL-18 and IL-1β were detected by ELISA,the mRNA expression of IL-17,TRAF6,NF-κB p65 and TGF-β1 in ovarian tissue were determined by RT-qPCR,the protein expression of TRAF6,NF-κB p65,TGF-β1,α-SMA,E-cadherin and the positive expression of IL-17RA,TRAF6,NF-κB p65,TGF-β1 and α-SMA in ovarian tissue were detected by Western blot and immunohistochemical staining respectively.Results Compared with the blank group,the body mass,ovarian mass and ovarian index significantly increased of rats in model group,with follicular cystic dilation,thinning of granulosa cell layer,and significant increase in fibrosis positive range(P<0.01),the serum contents of IL-17,IL-18 and IL-1β significantly increased(P<0.01),the expressions of IL-17,TRAF6,NF-κB p65,TGF-β1 mRNA and IL-17RA,TRAF6,NF-κB p65,TGF-β1 and α-SMA proteins significantly increased(P<0.01,P<0.05),the expression of E-cadherin protein significantly decreased(P<0.01).Compared with the model group,the ovarian index of rats in Yousiyue group and TCM medium-and high-dosage groups significantly decreased(P<0.01,P<0.05),the rats in TCM high-dosage group showed dominant follicles,increased thickness of granulosa cell layer,and significantly reduced fibrosis positive range(P<0.01),the serum contents of IL-17,IL-18 and IL-1β significantly decreased in Yousiyue group and TCM low-,medium-and high-dosage groups(P<0.01),the mRNA expressions of IL-17,TRAF6,NF-κB p65,TGF-β1,and the protein expressions of IL-17RA,TRAF6,NF-κB p65,TGF-β1 and α-SMA in ovarian tissue significantly decreaed in TCM high-dosage group(P<0.01,P<0.05),the expression of E-cadherin protein significantly increased(P<0.01).Conclusion Bushen Zhupai Decoction can decrease the body mass,improve the ovarian index,decrease the expressions of inflammatory factors and improve the status of ovarian fibrosis in PCOS rats.The mechanism may be related to the regulation of IL-17/TRAF6/NF-κB signaling pathway.
6.Research Progress in the Treatment of Non-alcoholic Fatty Liver Disease with Chinese Materia Medica Intervention of Ferroptosis
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(12):191-196
Ferroptosis is a unique iron-dependent cell death mode,which directly participates in and affects the occurrence,development and outcome of non-alcoholic fatty liver disease(NAFLD).Inhibiting ferroptosis can effectively prevent and improve liver injury in NAFLD patients.Chinese materia medica and the preparations have good inhibitory effect on liver ferroptosis and can play the role of liver protection.In this article,the pathogenesis of ferroptosis and its role in the evolution of NAFLD were reviewed,and the treatment of NAFLD by Chinese materia medica intervention in different pathways of ferroptosis mechanism was revealed,in order to provide reference for the mechanism research and clinical diagnosis and treatment of NAFLD.
7.Influence of correction degree on the clinical effect of valgus knee after total knee arthroplasty
Chao LI ; Mingliang JI ; Shanzheng WANG ; Wanshuang ZHANG ; Qing CHANG ; Jun LU ; Yonggang LI
Chinese Journal of Surgery 2021;59(12):1005-1011
Objective:To investigate the influence of correction degree on the clinical effect of valgus knee after total knee arthroplasty(TKA).Methods:Clinical data of 91 patients with unilateral valgus knee who underwent TKA from January 2015 to December 2018 at Department of Orthopaedic Surgery,Zhongda Hospital,Southeast University were retrospectively analyzed. According to postoperative hip-knee-ankle angle(HKA),these patients were divided into neutral group (-3°≤HKA≤3°) and residual valgus group(HKA>3°). There were 70 patients in the neutral group,with a median age of 71.5 years(range:57 to 91 years),and 21 patients in the residual valgus group,with a median age of 71.5 years(range:60 to 85 years). The postoperative clinical indicators,including the range of motion (ROM),American Knee Society score(KSS),Western Ontario and McMaster University(WOMAC) osteoarthritis index,forgotten joint score (FJS-12) were collected. Meanwhile,the imaging indexes of femoral angle,tibial angle,femoral prosthesis flexion angle,tibial prosthesis posterior slope angle,patella tilt angle,congruence angle were also collected. Additionally,postoperative complications were recorded. The clinical data were analyzed by independent sample t test,Mann-Whitney U test or χ 2 test. Results:There were no differences between the two groups in postoperative ROM (122.5°±8.5° vs. 119.4°±7.1°, t=1.534, P=0.128),KSS(159.8±8.9 vs. 150.0±8.9, t=-0.103, P=0.918),WOMAC osteoarthritis index(10.0(3.0) vs. 9.0(5.0), Z=-0.733, P=0.464),FJS-12(52.1(8.4) vs. 50.1(7.5), Z=-0.594, P=0.553)and femoral angle(94.0°(4.0°) vs. 94.0°(5.0°), Z=-0.209, P=0.835),tibial angle(90.0°(3.0°) vs. 91.0°(2.5°), Z=-1.226, P=0.220),femoral prosthesis flexion angle(3.0°(1.0°) vs. 3.0°(1.0°), Z=-0.652, P=0.514),tibial prosthesis posterior slope angle(4.0°(2.0°) vs. 4.0°(2.0°), Z=-0.763, P=0.445),patella tilt angle(9.0°(3.0°) vs. 9.0°(3.5°), Z=-1.429, P=0.153),congruence angle(14.0°(4.0°) vs. 15.0°(4.0°), Z=-1.690, P=0.091). However,abnormal proportions of patella tilt angle(33.3% vs.11.4%, χ2=5.630, P=0.018) and congruence angle(38.1% vs.17.1%, χ2=4.136, P=0.042) were apparent in the residual valgus group. The patients were follow-up for (32.1±5.2) months(range:24 to 40 months). Six weeks after operation,one case of joint stiffness occurred in the residual valgus group, which was improved after manual relaxation and rehabilitation exercises. At the last follow-up,two cases in the residual valgus group suffered patella instability which were fixed by patella braces and strengthening of the medial femoral muscles. There were no cases requiring reoperation for dislocation of patella,joint instability and aseptic loosening of the prosthesis in both groups. Conclusions:Residual valgus after TKA for valgus knee has no significant effect on the short-term clinical outcome,but it is more likely to cause abnormal proportions of patella tilt angle and congruence angle which may induce the risk of postoperative patella instability. The mid-and long-term outcomes need to be further investigated.
8.Influence of correction degree on the clinical effect of valgus knee after total knee arthroplasty
Chao LI ; Mingliang JI ; Shanzheng WANG ; Wanshuang ZHANG ; Qing CHANG ; Jun LU ; Yonggang LI
Chinese Journal of Surgery 2021;59(12):1005-1011
Objective:To investigate the influence of correction degree on the clinical effect of valgus knee after total knee arthroplasty(TKA).Methods:Clinical data of 91 patients with unilateral valgus knee who underwent TKA from January 2015 to December 2018 at Department of Orthopaedic Surgery,Zhongda Hospital,Southeast University were retrospectively analyzed. According to postoperative hip-knee-ankle angle(HKA),these patients were divided into neutral group (-3°≤HKA≤3°) and residual valgus group(HKA>3°). There were 70 patients in the neutral group,with a median age of 71.5 years(range:57 to 91 years),and 21 patients in the residual valgus group,with a median age of 71.5 years(range:60 to 85 years). The postoperative clinical indicators,including the range of motion (ROM),American Knee Society score(KSS),Western Ontario and McMaster University(WOMAC) osteoarthritis index,forgotten joint score (FJS-12) were collected. Meanwhile,the imaging indexes of femoral angle,tibial angle,femoral prosthesis flexion angle,tibial prosthesis posterior slope angle,patella tilt angle,congruence angle were also collected. Additionally,postoperative complications were recorded. The clinical data were analyzed by independent sample t test,Mann-Whitney U test or χ 2 test. Results:There were no differences between the two groups in postoperative ROM (122.5°±8.5° vs. 119.4°±7.1°, t=1.534, P=0.128),KSS(159.8±8.9 vs. 150.0±8.9, t=-0.103, P=0.918),WOMAC osteoarthritis index(10.0(3.0) vs. 9.0(5.0), Z=-0.733, P=0.464),FJS-12(52.1(8.4) vs. 50.1(7.5), Z=-0.594, P=0.553)and femoral angle(94.0°(4.0°) vs. 94.0°(5.0°), Z=-0.209, P=0.835),tibial angle(90.0°(3.0°) vs. 91.0°(2.5°), Z=-1.226, P=0.220),femoral prosthesis flexion angle(3.0°(1.0°) vs. 3.0°(1.0°), Z=-0.652, P=0.514),tibial prosthesis posterior slope angle(4.0°(2.0°) vs. 4.0°(2.0°), Z=-0.763, P=0.445),patella tilt angle(9.0°(3.0°) vs. 9.0°(3.5°), Z=-1.429, P=0.153),congruence angle(14.0°(4.0°) vs. 15.0°(4.0°), Z=-1.690, P=0.091). However,abnormal proportions of patella tilt angle(33.3% vs.11.4%, χ2=5.630, P=0.018) and congruence angle(38.1% vs.17.1%, χ2=4.136, P=0.042) were apparent in the residual valgus group. The patients were follow-up for (32.1±5.2) months(range:24 to 40 months). Six weeks after operation,one case of joint stiffness occurred in the residual valgus group, which was improved after manual relaxation and rehabilitation exercises. At the last follow-up,two cases in the residual valgus group suffered patella instability which were fixed by patella braces and strengthening of the medial femoral muscles. There were no cases requiring reoperation for dislocation of patella,joint instability and aseptic loosening of the prosthesis in both groups. Conclusions:Residual valgus after TKA for valgus knee has no significant effect on the short-term clinical outcome,but it is more likely to cause abnormal proportions of patella tilt angle and congruence angle which may induce the risk of postoperative patella instability. The mid-and long-term outcomes need to be further investigated.
9.Isolation, culture and identification of nucleus pulposus-derived mesenchymal stem cells from adult rats in vitro
Yunfeng RUI ; Shanzheng WANG ; Xinhui XIE ; Minghui SUN ; Yucheng LIN ; Gang LI ; Chen WANG
Chinese Journal of Tissue Engineering Research 2013;(49):8576-8582
BACKGROUND:Currently, cellular composition and the features of the nucleus pulposus are stil not to be clarified.
OBJECTIVE:To establish the in vitro culture system of rat nucleus pulposus-derived mesenchymal stem cells and to identify their multi-lineage differentiation potential.
METHODS:Mesenchymal stem cells from the nucleus pulposus tissues of Sprague-Dawley rats were cultured in vitro. Then, cells at passage 3 were induced to differentiate into osteoblasts, adipocytes and chondrocytes as experimental group. cells cultured with basic culture medium served as controls.
RESULTS AND CONCLUSION:cells isolated from rat nucleus pulposus could form the sunflower-like colonies and exhibit clone-like growth when they cultured at a low density. cells at passage 3 became homogeneous and exhibited fibroblast-like morphology. After 28 days of osteogenic induction, arizarin red positive signals were detected in the experimental group. The mRNA expressions of RunX2, osteopontin and osteocalcin were significantly increased in the experimental group, compared to the control group (P<0.05). After 21 days of adipogenic induction, oil red-O positive cells were detected in the experimental group. The mRNA expressions of C/EBPαand PPARγ2 were significantly increased in the experimental group, compared to the control group (P<0.05). After 21 days of chondrogenic induction, safranin O/fast green staining was positive in the experimental group. The mRNA expressions of aggrecan and Col2a1 were significantly increased in the experimental group, compared to the control group (P<0.05). Our findings in this study suggested that nucleus pulposus-derived mesenchymal stem cells could be isolated from the Sprague-Dawley rat nucleus pulposus and exhibited clonal-like growth when they were cultured in vitro. These cells were confirmed to have the potential to differentiate into adipocytes, osteoblasts and chondrocytes in vitro.
10.The clinical outcome of HLA haploidentical vs HLA-matched peripheral blood hematopoietic stem cell transplantation without in vitro T-cell depletion for malignant hematological diseases
Hailong YUAN ; Ming JIANG ; Bingzhao WEN ; Jianhua QU ; Ling LI ; Jianping HAO ; Yasen HALIDA ; Rong CHEN ; Xinhong GUO ; Shanzheng WANG ; Linglu DING
Chinese Journal of Organ Transplantation 2010;31(2):79-83
Objective To explore the clinical outcome of HLA haploidentical vs HLA-matcbed peripheral blood hematopoietic stem cell transplantation (PBSCT) without in vitro T-cell depletion for malignant hematological diseases. Methods 111 patients with malignant hematological diseases underwent PBSCT without in vitro T-cell depletion between May 2004 and February 2009, including 51 patients with HLA-haploidentical and 60 patients with HLA-matched. All patients have received myeloablative conditioning regimen. A two-agent based graft-versus-host disease (GVHD) prophylaxis was used as cyclosporine A and a short course of methotrexate. Mycophenolate mofetile was added for the patients with one locus mismatch. Mycophenolate mofetile, antithymocyte globulin and CD25 monoclonal antibody were added for the patients with 2-3 loci mismatch. The grafts were granulocyte colony-stimulating factor-mobilized peripheral blood stem cells without in vitro T-cell depletion. Results 111 patients achieved sustained and full donor-type engraftment. The median time to reach an absolute neutrophil count above 0.5×10~9/L was 14 days and that to a platelet count exceeding 20×10~9/L was 15 days in 51 HLA-haploidentical patients, and that was 12 days and 13 days in 60 HLA-matched patients, respectively. In 51 HLA-haploidentical patients, 25 patients developed aGVHD, including 20 cases of grade Ⅰ aGVHD, and 5 cases of grade Ⅱ. Thirty-three patients developed cGVHD with limited in 30 and extensive in 3. The 4-year cumulative incidence of cGVHD was 70.4 %. The 3-year probabilities of leukemia-free survival (LFS) were 74.5% (77.3 % for standard risk patients and 68.2 % for high risk patients respectively). Seven patients had recurrence. In 60 HLA-matched patients, 14 patients developed aGVHD, including 10 cases of grade Ⅰ, 2 cases of grade Ⅱ and 2 cases of grade Ⅲ. Thirty-seven patients developed cGVHD with limited in 32 and extensive in 5. The 4-year cumulative incidence of cGVHD was 58.1%. The 3-year probabilities of LFS were 72.1% (77.6 % for standard risk patients and 52.7 % for high risk patients respectively). Ten patients had recurrence. The incidence of aGVHD in HLA-haploidentical cohort was significantly higher than in HLA-matched cohort (P<0.05). There was no significant difference in incidence of cGVHD, incidence of relapse and LFS between HLA-haploidentical and HLA-matched cohorts (P>0.05). Conclusion Haploidentical PBSCT is feasible and safe for malignant hematological diseases to use myeloablative conditioning regimen in combination with intensive immunosuppressants without in vitro T cell depletion.

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