[Efficacy and safety analysis of venetoclax combined with hypomethylating agents for the treatment of higher-risk myelodysplastic syndromes in the real world].

Objective: To investigate the efficacy and safety of combining venetoclax (VEN) with hypomethylated drugs (HMA) in the treatment of higher-risk (IPSS-R score >3.5) myelodysplastic syndromes (MDS) . Methods: From March 2021 to December 2022, forty-five MDS patients with intermediate and high risk were treated with VEN in combination with HMAs. Clinical data were collected and analyzed retrospectively, including gender, age, MDS subtype, IPSS-R score, treatment regimen, and efficacy, etc. Kaplan-Meier method and Cox regression model were used to analyze univariate and multivariate of survival prognosis. Results: ①Forty-five patients with MDS, including ninety-one percent were classified as high or very high risk. According to the 2023 consensus proposal for revised International Working Group response criteria for higher-risk MDS, the overall response rate (ORR) was 62.2% (28/45), with the complete response rate (CR) was 33.3% (15/45). For twenty-five naïve MDS, the ORR was 68% (17/25) and the CR rate was 32% (8/25). In nonfirst-line patients, the ORR and CR were 55% (11/20) and 35% (7/20) respectively. The median cycle to best response was 1 (1-4). ②With a median followup of 189 days, the median overall survival (OS) time was 499 (95% confidence interval, 287-711) days, and most patients died from disease progression. Responders had a significantly better median OS time than nonresponders (499 days vs 228 days, P<0.001). Multifactor analysis revealed that IPSS-R score and response to treatment were independent prognostic factors for OS; the presence of SETBP1 gene mutations was associated with a longer hospital stay (51.5 days vs 27 days, P=0.017) . Conclusions: There is clinical benefit of venetoclax in combination with hypomethylated agents in patients with higher-risk MDS, but adverse events such as severe hypocytopenia during treatment should be avoided.

[Molecular features of 109 patients with chronic myelomonocytic leukemia in a single center].

Objective: To explore the molecular features of chronic myelomonocytic leukemia (CMML) . Methods: According to 2022 World Health Organization (WHO 2022) classification, 113 CMML patients and 840 myelodysplastic syndrome (MDS) patients from March 2016 to October 2021 were reclassified, and the clinical and molecular features of CMML patients were analyzed. Results: Among 113 CMML patients, 23 (20.4%) were re-diagnosed as acute myeloid leukemia (AML), including 18 AML with NPM1 mutation, 3 AML with KMT2A rearrangement, and 2 AML with MECOM rearrangement. The remaining 90 patients met the WHO 2022 CMML criteria. In addition, 19 of 840 (2.3%) MDS patients met the WHO 2022 CMML criteria. At least one gene mutation was detected in 99% of CMML patients, and the median number of mutations was 4. The genes with mutation frequency ≥ 10% were: ASXL1 (48%), NRAS (34%), RUNX1 (33%), TET2 (28%), U2AF1 (23%), SRSF2 (21.1%), SETBP1 (20%), KRAS (17%), CBL (15.6%) and DNMT3A (11%). Paired analysis showed that SRSF2 was frequently co-mutated with ASXL1 (OR=4.129, 95% CI 1.481-11.510, Q=0.007) and TET2 (OR=5.276, 95% CI 1.979-14.065, Q=0.001). SRSF2 and TET2 frequently occurred in elderly (≥60 years) patients with myeloproliferative CMML (MP-CMML). U2AF1 mutations were often mutually exclusive with TET2 (OR=0.174, 95% CI 0.038-0.791, Q=0.024), and were common in younger (<60 years) patients with myelodysplastic CMML (MD-CMML). Compared with patients with absolute monocyte count (AMoC) ≥1×10(9)/L and <1×10(9)/L, the former had a higher median age of onset (60 years old vs 47 years old, P<0.001), white blood cell count (15.9×10(9)/L vs 4.4×10(9)/L, P<0.001), proportion of monocytes (21.5% vs 15%, P=0.001), and hemoglobin level (86 g/L vs 74 g/L, P=0.014). TET2 mutations (P=0.021) and SRSF2 mutations (P=0.011) were more common in patients with AMoC≥1×10(9)/L, whereas U2AF1 mutations (P<0.001) were more common in patients with AMoC<1×10(9)/L. There was no significant difference in the frequency of other gene mutations between the two groups. Conclusion: According to WHO 2022 classification, nearly 20% of CMML patients had AMoC<1×10(9)/L at the time of diagnosis, and MD-CMML and MP-CMML had different molecular features.

[Risk factors for leukemia transformation in patients with myelodysplastic syndromes].

Objective: To explore the risk factors in leukemia transformation (LT) in those with myelodysplastic syndromes (MDS) . Methods: From January 2012 to December 2020,data on 320 patients with newly diagnosed primary MDS were gathered from the MDS center. The clinical features and molecular characteristics are explored. Additionally, a retrospective analysis of risk factors for the development of acute leukemia from MDS was done. Results: The median follow-up was13.6 (0.4-107.3) months. 23.4% (75/320) of the MDS patients had LT group. Significant differences between the LT group and non-LT group can be seen in age (P<0.001) , bone marrow blast percentage (P<0.001) , bone marrow fibrosis (P=0.046) , WHO classification (P<0.001) , IPSS-R (P<0.001) and IPSS-R karyotype group (P=0.001) . The median number of mutation of LT group was 1 (1, 3) , that in non-LT group was 1 (0, 2) ,which had a statistical difference (P=0.003) .At the time of the initial diagnosis of MDS, the LT group had higher rates of the TP53 mutation (P=0.034) , DNMT3A mutation (P=0.026) , NRAS mutation (P=0.027) and NPM1 mutation (P=0.017) . Compared with the mutations at first diagnosis and LT of six patients, the number of mutations increased and the variant allele frequencies (VAF) increased significantly in LT patients. Higher bone marrow blast percentage (Refer to <5% , 5% -10% : HR=4.587, 95% CI 2.214 to 9.504, P<0.001, >10% : HR=9.352, 95% CI 4.049 to 21.600, P<0.001) , IPSS-R cytogenetic risk groups (HR=2.603, 95% CI 1.229-5.511, P=0.012) , DNMT3A mutation (HR=4.507, 95% CI 1.889-10.753, P=0.001) , and NPM1 mutation (HR=3.341, 95% CI 1.164-9.591, P=0.025) were all independently associated with LT in MDS patients, according to results of multivariate Cox regression. Conclusion: Bone marrow blast percentage, IPSS-R cytogenetic risk groups, DNMT3A mutation, and NPM1 mutation are independent risk factors in LT for MDS patients.

[Prognostic factors of cyclosporine A combined with danazol with or without thalidomide in myelodysplastic syndrome treatment with low-percentage bone marrow blasts].

Objective: To explore the outcome of cyclosporine A (CsA) combined with danazol with or without thalidomide regimen for myelodysplastic syndrome (MDS) with low-percentage bone marrow blasts and predictive factors for treatment response. Methods: Data of 115 subjects who were newly diagnosed with primary MDS with low-percentage bone marrow blasts and were treated with CsA combined with danazol with or without thalidomide from December 2011 to December 2019 in our center were collected. Their clinical features, efficacy, and predictive factors of efficacy were retrospectively analyzed. A model for predicting this response was developed. Results: A total of 55 subjects responded (47.8%) , including 11 complete responses and 44 hematologic improvements. Fifty-two patients (52/105, 49.5%) achieved erythrocyte response; 35 (35/86, 40.7%) , platelet response; and 14 (14/40, 35%) , neutrophil response. Of 29 subjects (24.1%) , 7 who were red blood cell (RBC) transfusion-dependent became independent of transfusion. The median response duration was 20 months (range, 3-84 months) . In the univariate analysis, patients <0 years had a higher response rate than those ≥60 years (52.5% vs 22.2%, P=0.018) . Contrarily, the response rate was substantially decreased in patients with RBC transfusion dependence compared with those without RBC transfusion dependence (24.1% vs 55.8%, P=0.003) , as well as in patients with the mutated U2AF1 compared with those with the wild-type U2AF1 (26.1% vs 53.2%, P=0.020) . In multivariable analyses, age <0 years (OR=4.302, 95% CI 1.245-14.820, P=0.021) , RBC transfusion dependence (OR=3.774, 95% CI 1.400-10.177, P=0.009) , and U2AF1 mutation (OR=3.414, 95% CI 1.168-9.978, P=0.025) were significantly correlated with response. Variables that independently predicted the response were combined to generate the predictive model. According to the model, the overall response rates of patients with 0, 1, 2, and 3 risk factors were 65%, 30%-35%, 10%-15%, and 3%, respectively. Conclusion: CsA combined with danazol with or without thalidomide regimen could improve cytopenia symptoms in patients with MDS with low-percentage bone marrow blasts. At age <60 years, no transfusion dependence of RBC and wild-type U2AF1 mutation is a favorable prognostic factor.

[The observation of secretogranin â ¢ in the peripheral blood and vitreous of patients with diabetic retinopathy].

Objective: To study the expression levels of secretogranin Ⅲ (SCG3) in the peripheral blood and vitreous of patients with diabetic retinopathy (DR). Methods: Cross-sectional research. A total of 77 patients (41 men and 36 women, 77 eyes) received vitrectomy in Tianjin Medical University Eye Hospital from May to December 2018, with an average age of (60.75±11.34) years. According to the blood glucose level, diabetes history and fundus status, all the patients were divided into the DR group and the non-diabetic group. According to the patients' blood lipids and body mass index (BMI), patients were further divided into subgroups of high blood lipids, normal blood lipids, high BMI and normal BMI. All patients were tested with eye examinations, height and weight to calculate the BMI, and blood lipid levels in the peripheral blood. The vitreous was collected during the vitrectomy surgery, and the levels of SCG3 in the vitreous and peripheral blood were analyzed by ELISA. All the data were analyzed statistically with Wilcoxon rank sum test. Results: There were 43 patients in the DR group, among whom 25 had hyperlipidemia, 18 had normal blood lipids, 22 had a high BMI, and 21 had a normal BMI. There were 34 patients in the non-diabetic group, among whom 13 had hyperlipidemia, 21 had normal blood lipids, 17 had a high BMI, and 17 had a normal BMI. The level of SCG3 in the DR group [6.02 (4.34, 11.76) ng/ml] was higher than that in the non-diabetic group [4.30 (3.20, 10.78) ng/ml] (Z=-2.339, P =0.019). The level of SCG3 in the hyperlipidemia subgroup of the DR patients [7.94 (5.33, 13.51) ng/ml] was higher than that in the normal blood lipid subgroup of the non-diabetic patients [4.04 (3.12, 7.77) ng/ml] (Z=-3.473, P=0.001), and higher than that in the DR patients without hyperlipidemia [4.45 (3.71, 9.14) ng/ml] (Z=-2.511, P=0.012). The level of SCG3 in the DR patients with a high BMI [7.12 (4.56, 13.12) ng/ml] was higher than that in the non-diabetic patients with a normal BMI [3.53 (3.16, 4.38) ng/ml] (Z=-3.767, P =0.000). The level of SCG3 in the DR patients with a normal BMI [5.72 (4.10, 11.60) ng/ml] was higher than that in the non-diabetic patients with a normal BMI (Z=-2.862, P = 0.004). SCG3 in the plasma was rare or can not be detected. Conclusions: The concentration of SCG3 in the vitreous increase in DR patients. However, SCG3 can not be detected in the healthy vascular system. (Chin J Ophthalmol, 2020, 56: 933-937).

Oncogene miR-934 promotes ovarian cancer cell proliferation and inhibits cell apoptosis through targeting BRMS1L.

OBJECTIVE: Ovarian cancer is a common malignant cancer among women. Increasing studies have demonstrated that microRNAs function as important regulation factors in the progression of ovarian cancer. MATERIALS AND METHODS: Human ovarian cancer cell lines HO8910 and OVCAR-3 were transfected with miR-934 inhibitor and corresponding negative control (inhibitor control). Cell proliferation and apoptosis were detected by cell counting kit-8 (CCK-8) and TUNEL assay, respectively. The expression levels of proliferation/apoptosis-related genes and BRMS1L were measured by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blotting. Furthermore, the association between miR-934 and BRMS1L was investigated through luciferase reporter assays. RESULTS: MiR-934 was significantly increased in ovarian cancer cell lines, whereas BRMS1L was significantly decreased. Downregulated miR-934 remarkably inhibited cell proliferation and induced cell apoptosis. Meanwhile, miR-934 could influence the expression levels of Ki67, Cyclin D1, Caspase3, and Bcl-2. In addition, BRMS1L was identified as a target gene of miR-934. CONCLUSIONS: Oncogene miR-934 promotes ovarian cancer cell proliferation and inhibits cell apoptosis through targeting BRMS1L. MiR-934 and BRMS1L may be novel biomarkers or therapeutic targets for ovarian cancer in the future.

[Long term clinical observation on postoperative microstructural change of macula lutea in treatment of idiopathic macular epiretinal membrane].

Objective: To evaluate the relationship between the structure of macular region and prognosis after vitrectomy in idiopathic macular epiretinal membrane patients. Methods: Retrospective case series study. The cases of 74 idiopathic macular epiretinal membrane patients (31 female patients, 43 male patients, aged 68.1±6.9) who received surgery treatment in our hospital during January 2013 and December were collected and analyzed. Ophthalmic routine examination and optical coherence tomography (OCT) have been conducted for all patients prior to operation. All eyes underwent vitecrtomy and macular epiretinal membrane peeling. During the 12 months follow up, the preoperative and postoperative data including best corrected visual acuity (BCVA), foveal thickness, changes of morphology in ellipsoid zone were observed and analyzed. All the patients were divided into two groups, continuous group (52 patients) and discontinuous group (22 patients), based on the preoperative continuity status of ellipsoid zone. The data of preoperative macular retinal thickness, the relationship between age and course of the disease, preoperative BCVA and postoperative BCVA were analyzed statistically with repeated ANOVA method. The postoperative BCVA were analyzed statistically with t test mtehod, based on the different grouping of continuity status of ellipsoid zone. Results: The preoperative LogMar BCVA, and the postoperative LogMAR BCVA of 1 month, 3 month, 6 month and 12 month after operation were 0.59±0.26, 0.63±0.34, 0.46±0.22, 0.45±0.23, 0.41±0.23 (F=25.122, P<0.05) respectively. The preoperative foveal thickness, and the postoperative foveal thickness of 1 month, 3 month, 6 month and 12 month after operation were (468.12±73.07), (371.57±57.09), (320.57±65.88), (294.85±69.36), (283.5±66.56)µm (F=8.802, P<0.05), the difference between the figures are of statistical significance. The postoperative BCVA of continuous group (0.34±0.27) improved significantly, as compared to the BCVA of discontinuous group (0.62±0.24) (t=-4.209, P<0.05). There is no obvious relationship between the degree of preoperative macular edema and postoperative visual function (r=0.015, P> 0.015). Seven patients still have macular edema one year after surgery. Conclusion: The postoperative visual function in patients with idiopathic macular epiretinal membrane has negative correlation with preoperative thickness of fovea, and has positive correlation with continuity of ellipsoid zone. (Chin J Ophthalmol, 2018, 54: 258-262).

A rapid review of needs assessment tools for post-treatment cancer survivors.

Relevant, comprehensive and psychometrically rigorous needs assessment tools are needed to ensure appropriate care is delivered to cancer survivors who have completed treatment. The aim of this rapid review was to identify and describe needs assessment tools that are used in cancer survivors post-treatment, assess their psychometric properties and describe their use in clinical care. The electronic databases Medline, Cochrane Library, CINAHL and PsycINFO were searched. Six studies were identified that described five needs assessment tools used in cancer survivors post-treatment. None of these tools covered all domains of unmet need nor demonstrated adequate evidence of all recommended criteria of validity and reliability. Few had been evaluated for use in a clinical environment. Out of the five tools, the Survivor Unmet Needs Survey (SUNS) showed the strongest psychometric properties. There is little empirical evidence available to guide recommendations on the most appropriate process of conducting needs assessment with cancer survivors once they have completed treatment.

Effect of the acrosome reaction on the efficiency of sperm-mediated DNA transfer.

Sperm-mediated gene transfer (SMGT) is based on the ability of spermatozoa to bind exoge- nous DNA and transfer it into oocytes by fertilization. However, SMGT is still undergoing opti- mization to improve its efficiency to produce transgenic animals. The acrosome reaction is neces- sary for spermatozoa to carry the exogenous DNA into oocytes. In this study, the effect of the acrosome reaction on the efficiency of spermatozoa carrying exogenous DNA was evalua- ted. The results showed that the efficiency of the acrosome reaction was significantly higher (p⟨0.05) after incubation with 50 µmol/L progesterone compared to incubation without proges- terone. It was significantly higher (p⟨0.05) in the 20, 40, and 60 min of progesterone treatment groups than in the 0 min treatment group. The spermatozoa were further incubated with cyanine dye Cy5 labeled DNA (Cy5-DNA) for 30 min at 37°C, and positive fluorescence signals were detected after the acrosome reaction was induced by progesterone at concentrations of 0 and 50 µmol/L for 40 min. The percentage of positive Cy5-DNA signals in spermatozoa was 96.61±2.06% and 97.51±2.03% following exposure to 0 and 50 µmol/L progesterone, respective- ly. The percentage of partial spermatozoa heads observed following combination with Cy5-DNA was 39.73±3.03% and 56.88±3.12% following exposure to 0 and 50 µmol/L progesterone, respec- tively. The ratio of positively stained spermatozoa combined with exogenous DNA showed no reduction after the acrosome reaction. These results suggest that the acrosome reaction might not be the key factor affecting the efficiency of SMGT.

CUL4B promotes gastric cancer invasion and metastasis-involvement of upregulation of HER2.

Cullin 4B (CUL4B) is a scaffold protein overexpressed in several solid malignancies. It is known to silence tumor suppressor through post-transcriptional manner. However, its clinical significance and underlying molecular mechanisms in gastric cancer (GC) remain largely unknown. In this study, we found that CUL4B was significantly overexpressed in GC tissues and its overexpression was correlated with lymph node metastasis and poor prognosis. Through gain- and loss-of-function experiments, we showed that CUL4B promotes GC cell invasion and epithelial-mesenchymal transition (EMT) in vitro, as well as tumor growth and metastasis in vivo. Mechanistically, we identified HER2 as a downstream target gene of CUL4B in GC. CUL4B unregulated HER2 expression via transcriptionally repressing miR-125a. Intriguingly, HER2 inhibitors significantly reversed CUL4B-induced EMT in vitro and partially blocked GC metastasis in tumor xenografts with CUL4B overexpression. Finally, we suggested the involvement of the PI3K/AKT pathway in CUL4B-induced HER2 upregulation in GC. In all, we proposed a model for a CUL4B-miR-125a-HER2 oncoprotein axis, which provided novel insight into how HER2 was activated and contributed to GC progression and metastasis.

[Cellular and molecular mechanisms of anti-inflammatory effect of peroxisome proliferator-activated receptor α].

Objective: To investigate the cellular and molecular mechanisms of the anti-inflammatory effect of peroxisome proliferator-activated receptor α (PPARα). Methods: Firstly, bone marrow-derived macrophages (BMDMs) were randomly divided into control group, LPS group, WY14643 10 µmol/L group, WY14643 25 µmol/L group, and WY14643 50 µmol/L group using a random number table. Secondly, BMDMs were randomly divided into LPS group, WY14643+LPS group, and 3-MA+WY14643+LPS group. Primary BMDMs were stimulated by LPS (20 ng/ml) to establish the cellular model of inflammation. The selective agonist of PPARα WY14643 was administered at doses of 10, 25, and 50 µmol/L (50 µmol/L for the second part of the experiment) at 2 hours before model establishment. The autophagy inhibitor 3-MA was administered at a dose of 10 mmol/L at 2 hours before model establishment. The cells in the control group were treated with dimethylsulfoxide (DMSO) at the same dose. The cells were transfected with GFP-LC3 plasmids at 24 hours before model establishment. The cells were harvested at 6 hours after LPS stimulation and related tests were performed. Green fluorescent protein was measured under a fluorescence microscope to evaluate autophagy activity. Quantitative real-time PCR was used to measure tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and mRNA expression of chemokine-1 (CXCL-1) and chemokine-10 (CXCL-10). Western blot was used to measure PPARα and autophagy-related proteins LC3, ATG-5, ATG-7, and LAMP-1. A one-way analysis of variance was used for comparison between groups, and the LSD-t test was used for comparison between any two groups. Results: In vitro, PPARα activation inhibited LPS-induced inflammatory response in primary macrophages in a dose-dependent manner. The results of gene expression showed that the relative expression of TNF-α, IL-1ß, IL-6, CXCL-1, and CXCL-10 was as follows in the control group, LPS group, WY14643 10 µmol group, WY14643 25 µmol group, and WY14643 50 µmol group: TNF-α (0.085±0.009, 4.065±0.544, 3.281±0.368, 1.780±0.293, and 0.781±0.303, P < 0.01), IL-1ß (0.081±0.017, 0.776±0.303, 0.225±0.154, 0.161±0.068, and 0.101±0.025, P < 0.05), IL-6 (0.041±0.011, 0.189±0.014, 0.144±0.033, 0.126±0.013, and 0.048±0.015, P < 0.01), CXCL-1 (0.051±0.011, 0.515±0.145, 0.356±0.078, 0.257±0.068, and 0.069±0.030, P < 0.01), and CXCL-10 (0.126±0.068, 0.831±0.093, 0.508±0245, 0.474±0.047, and 0.204±0.021, P < 0.05). In vitro, PPARα activation promoted autophagy in vitro in a dose-dependent manner. The results of Western blot and fluorescence microscopy in the control group, LPS group, WY14643 10 µmol group, WY14643 25 µmol group, and WY14643 50 µmol group showed that the expression of autophagy-related proteins and autophagosome formation gradually increased with the increasing concentration of WY14643. In vitro, WY14643 inhibited autophagy, promoted inflammatory response in primary macrophages, and reversed the anti-inflammatory effect of PPARα. The results of gene expression showed that the relative expression of TNF-α, IL-1ß, IL-6, CXCL-1, and CXCL-10 was as follows in the LPS group, WY14643+LPS group, and 3-MA+WY14643+LPS group: TNFα (4.327±0.478, 1.218±0.424, and 3.901±0.447, P < 0.05), IL-1ß (4.277±0.407, 1.418±0.424, and 3.029±0.192, P < 0.01), IL-6 (4.175±0.549, 1.373±0.499, and 4.031±0.475, P < 0.05), CXCL-1 (8.199±1.149, 2.024±0.547, and 5.973±0.843, P < 0.05), and CXCL-10 (1.208±0.148, 0.206±0.069, and 0.798±0.170, P < 0.05). Conclusion: PPARα can promote cell autophagy and inhibit inflammatory response and may become a new therapeutic target for clinical prevention and treatment of inflammatory disease.

Detection of serum antinuclear antibodies in lymphoma patients.

We investigated the presence of serum antinuclear antibodies (ANAs) and autoantibodies and their relationship with serum prognostic indicators in lymphoma patients. The study population comprised 127 patients diagnosed with lymphoma and 138 healthy control subjects. The blood samples of the participants were assayed for ANAs by immunofluorescence, and autoantibodies were detected by western blotting. Serum ANAs were detected in 31.5 (40/127) and 6.5% (9/138) of lymphoma patients and control subjects, respectively. There was a statistically significant difference between the lymphoma and the control groups (P < 0.05). The level of lactate dehydrogenase in the ANA-positive subjects was significantly lower than in the ANA-negative subjects (P < 0.05). Low ANA titers (1:100) were commonly found in the ANA-positive subjects and the control subjects, and the fluorescence models were diverse. Autoantibodies were found in 35% (14/40) of the ANA-positive patients by western blotting. Detection of ANAs in lymphoma patients helps in determining the diagnosis and prognosis of lymphoma, but has no independent diagnostic value; there are still various autoantibodies of unknown significance that require further study.

Peroxisome proliferator-activated receptor α activation attenuates the inflammatory response to protect the liver from acute failure by promoting the autophagy pathway.

Peroxisome proliferator-activated receptor α (PPARα) has been reported to induce a potent anti-inflammatory response. Autophagy is a recently recognized rudimentary cellular response to inflammation and injury. The aim of the present study was to test the hypothesis that PPARα activation mediates autophagy to inhibit liver inflammation and protect against acute liver failure (ALF). PPARα expression during ALF and the impact of PPARα activation by Wy-14 643 on the hepatic immune response were studied in a D-galactosamine/lipopolysaccharide-induced mouse model. Autophagy was inhibited by 3-methyladenine or small interfering RNA (siRNA) against Atg7. In both the mouse model and human ALF subjects, PPARα was significantly downregulated in the injured liver. PPARα activation by pretreatment with Wy-14 643 protected against liver injury in mice. The protective effect of PPARα activation relied on the suppression of inflammatory mechanisms through the induction of autophagy. This hypothesis is supported by the following evidence: first, PPARα activation suppressed proinflammatory responses and inhibited phosphorylated NF-κBp65, phosphorylated JNK and phosphorylated ERK pathways in vivo. Second, protection by PPARα activation was due to the induction of autophagy because inhibition of autophagy by 3-methyladenine or Atg7 siRNA reversed liver protection and inflammation. Third, PPARα activation directly induced autophagy in primary macrophages in vitro, which protected cells from a lipopolysaccharide-induced proinflammatory response. Here, for the first time, we have demonstrated that PPARα-mediated induction of autophagy ameliorated liver injury in cases of ALF by attenuating inflammatory responses, indicating a potential therapeutic application for ALF treatment.

Tropomyosin is localized in the nuclear matrix and chromosome scaffold of Physarum polycephalum.

The nuclei and chromosomes were isolated from plasmodia of Physarum polycephalum. The nuclear matrix and chromosome scaffold were obtained after the DNA and most of the proteins were extracted with DNase I and 2 M NaCl. SDS-PAGE analyses revealed that the nuclear matrix and chromosome scaffold contained a 37 kD polypeptide which is equivalent to tropomyosin in molecular weight. Immunofluorescence observations upon slide preparations labeled with anti-tropomyosin antibody showed that the nuclear matrix and chromosome scaffold emanated bright fluorescence, suggesting the presence of the antigen in them. Immunodotting results confirmed the presence of tropomyosin in the nuclear matrix and chromosome scaffold. Immunoelectron microscopic observations further demonstrated that tropomyosin was dispersively distributed in the interphase nuclei and metaphase chromosomes.

[Immunocytochemical identification of tropomyosin in nucleoli and nucleolar matrix of Physarum polycephalum].

Nucleoli were isolated from physarum polycephalum, and nucleolar matrix was prepared by digesting the nucleoli respectively with DNase 1, 0.25 mol/L (NH4)2SO4 and 2 mol/L NaCl to remove DNA and most proteins. SDS-PAGE analysis indicated that there were about 20 polypeptides in nucleolar matrix component, including the 37 kD polypeptide which was similar to tropomyosin in molecular weight. The result of indirect immunofluorescence treated with anti-tropomyosin antibody and sheep anti-rabbit IgG antibody labelled with FITC showed that bright fluorescence was observed in the nucleoli and nucleolar matrix, but no bright fluorescence in the controls. Indirect Immunoblotting detection further verified that tropomyosin existed in nucleolar matrix. Protein A-colloidal gold immunoelectron microscopic study showed that there were many gold particles in the specimens labelled with tropomyosin antibody, and there were few gold particles found in the controls. Tropomyosin distributed dispersedly in nucleoli.