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.

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.

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.

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.

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.

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.

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.

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.

BACKGROUND: Lack of human leucocyte antigen-matched family donors has restricted the application of hematopoietic cell transplantation. Due to immunological disorder of humam leucocyte antigen misfit, common way for haploidentical transplantation is associated with poor engraftment and severe graft-versus-host disease. Because not every patient has HLA-Jdentical family member, a substantial proportion of patients will receive haploidentical transplantation. OBJECTIVE: To explore the curative effect on malignant hematological diseases of haploidentical peripheral blood stem cell transplantation (PBSCT) using myeloablative conditioning regimen in combination of proper immunosuppressants without in vitro T-cell depletion. DESIGN, TIME AND SETTING: A case observation was performed at the Department of Hematology in the First Affiliated Hospital of Xinjiang Medical University from July 2002 to June 2008. PARTICIPANTS: Forty-two patients with malignant hematological diseases, including 29 standard-risk patients and 13 high-risk patients, age from 10 to 48 years, were transplanted with cells from a haploidentical family donor with 1-3 mismatched loci of HLA antigens. Seven patients had 1 locus mismatched donors and thirty-five patients had 2-3 loci mismatched donors. METHODS: The patients have received myeloablative conditioning regimen. A two-agent based graft-versus-host disease (GVHD) prophylaxis was used as cyclospodne A and a short course of methotrexate. Mycophenolate mofetile was added for 1 locus mismatched patients. Mycophenolate mofetile, antithymocyte globulin and CD25 mono-colonal antibody were added for 2-3 loci mismatched patients. The grafts were granulocyte colony-stimulating factor-mobilized peripheral blood stem cells without in vitro T-cell depletion. MAIN OUTCOME MEASURES: Engraftment, GVHD incidence and severity, relapse and leukemia-free survival and the immune function of patients in months 1, 3, 6, 12 and 18 postoperatively. RESULTS: Totally 42 patients achieved complete and sustained donor-type engraftment. Nineteen patients developed acute GVHD, the 2-year cumulative incidences of acute GVHD were 50.8%, gradeⅠ acute GVHD occurred in 16 cases and grade Ⅱ in 3 cases. Thirty-one patients were followed up more than 6 months, 23 of them developed chronic GVHD with limited in 20 and extensive in 3, the 2-year cumulative incidences of chronic GVHD were 57.1%. No patients died of GVHD. There were no significant differences in the reduction and recovery of T cells and B cells between HLA haploidentical PBSCT without in vitro T cell depletion and HLA-matched PBSCT. CONCLUSION: Haploidentical PBSCT is feasible and safe for malignant hematological diseases to use myeloablative conditioning regiment combination of intensive immunosuppressants without in vitro T cell depletion. A large amount of clinical cases need to be investigated in the near future.

Objective To investigate the expression of VEGF-D and MMP-2 and explore their relationship with tumor invasion and local metastasis in pancreatic carcinoma.Methods The expression of VEGF-D and MMP-2 in 30 patients with pancreatic carcinomas and 6 cases with normal pancreatic tissues were detected by SABC immunohistochemical method.Results The positive rates of VEGF-D and MMP-2 proteins were 60%(18/30)and 66.7%(20/30)respectively,no expression of VEGF-D and MMP-2 was detected in normal pancreatie tissues.The expression of VEGF-D was significantly correlated with lymph node metastasis(P＜0.05),and the expression of MMP-2 was significantly correlated with lymph node metastasis and tumor local invasion(P＜0.05).The expression of VEGF-D protein Was positively correlated with MMP-2 protein(rs=0.54,P＜0.01).Conclusions There is a significantly positive correlation between high expression of VEGF-D and MMP-2 proteins in pancreatic carcinoma,and they may contribute to the invasive properties of pancreatic cancer.