Comparison of immune responses of acellular vascular materials prepared by the two methods in animals / 中国组织工程研究

BACKGROUND: As a mature method of decellularization at present, descaling agent has the advantages of simple operation, small damage, and easy control of experimental conditions. Some studies have combined different descaling agents to achieve good results in decellularization. OBJECTIVE: To compare the effect of two descaling agents and the immune response of two kinds of acellular vascular materials subcutaneously implanted into animals. METHODS: Two methods were used to treat the porcine carotid artery for descaling agent. One group was treated in the mixed solution of 1% sodium dodecyl sulfate and 1% sodium deoxycholate for 72 hours; the other group was treated in the mixed solution of 1% sodium dodecyl sulfate and 1% Triton X-100 for 72 hours. After cell removal, histological analysis, scanning electron microscope analysis, mechanical analysis and DNA content detection were carried out. Two groups of acellular vascular materials were implanted subcutaneously in the back of SD rats. After 1, 2, 4 and 8 weeks, the grafted vascular materials were taken out for hematoxylin and eosin and immunofluorescence staining. This study was approved by the Animal Ethics Committee of Jiangnan University. RESULTS AND CONCLUSION: (1) Histological analysis showed that 1% sodium dodecyl sulfate combined with 1% sodium deoxycholate effectively removed the cell components and retained the extracellular matrix structure, and the effect of cell removal was better than 1% sodium dodecyl sulfate combined with 1% Triton X-100. (2) The DNA content of 1% sodium dodecyl sulfate combined with 1% sodium deoxycholate group was significantly lower than that of 1% sodium dodecyl sulfate combined with 1% Triton X-100 group (P 0.05). (3) Scanning electron microscopy showed that 1% sodium dodecyl sulfate combined with 1% Triton X-100 significantly damaged the extracellular matrix compared with 1% sodium dodecyl sulfate combined with 1% sodium deoxycholate. (4) In the subcutaneous implantation experiment, a large number of inflammatory cells infiltrated around the carotid artery materials of the two groups at 1 week after operation. No inflammatory cells infiltrated around the acellular materials of 1% sodium dodecyl sulfate and 1% sodium deoxycholate at 8 weeks after operation. Lymphocyte infiltrated around the acellular materials of 1% sodium dodecyl sulfate and 1% Triton X-100 group. 1% sodium dodecyl sulfate combined with 1% Triton X-100 acellular material mainly induced macrophages to differentiate into M1 type. 1% sodium dodecyl sulfate combined with 1% sodium deoxycholate acellular material mainly induced macrophages to differentiate into M2 type. (5) Compared with 1% sodium dodecyl sulfate and 1% Triton X-100, 1% sodium dodecyl sulfate and 1% sodium deoxycholate are more promising cell-free treatment.

The diagnostic benefit of antibodies against ribosomal proteins in systemic lupus erythematosus

Abstract Background Anti-ribosomal P (anti-Rib-P) antibody is a specific serological marker for systemic lupus erythematosus (SLE) and routinely tested by targeting the common epitope of three ribosomal proteins of P0, P1 and P2. This study aimed to investigate if testing antibodies against individual ribosomal protein, but not the common epitope, is required to achieve the best diagnostic benefit in SLE. Methods The study included 82 patients with SLE and 22 healthy donors. Serum antibodies were determined by ELISA and immunoblot. Results The prevalence of each antibody determined by ELISA was 35.4% (anti-Rib-P), 45.1% (anti-Rib-P0), 32.9% (anti-Rib-P1) and 40.2% (anti-Rib-P2) at 99% specificity, respectively. Of 53 patients with negative anti-Rib-P antibody, 21 (39.6%) were positive for anti-Rib-P0, 9 (17.0%) for anti-Rib-P1 and 12 (22.6%) for anti-Rib-P2 antibody. The positive rate of anti-Rib-P antibody detected by ELISA was close to the results by immunoblot (33.4%). Patients with any of these antibodies were featured by higher disease activity and prevalence of skin rashes than those with negative antibodies. Moreover, each antibody was particularly related to some clinical and laboratory disorders. The distribution of subclasses of IgG1-4 was varied with each antibody. Anti-Rib-P0 IgG1 and IgG3 were strongly correlated with disease activity and lower serum complement components 3 and 4. Conclusions Anti-Rib-P antibody is not adequate to predict the existence of antibodies against ribosomal P0, P1 and P2 protein. The examination of antibodies against each ribosomal protein is required to achieve additional diagnostic benefit and to evaluate the association with clinical and serological disorders as well.(AU)

GADD45g exerts anti-tumor effect in acute myeloid leukemia via inhibition of E2F1 / 中国肿瘤生物治疗杂志

@# Objective: To investigate the correlation between the expression of E2F1 and growth arrest and DNA damage inducible protein 45g (GADD45g) in patients with acute myeloid leukemia (AML), and to explore whether GADD45g exerts its induction of DNA damage, cell apoptosis, senescence, cell cycle arrest and drug sensitivity in AML through inhibition of E2F1. Methods: A total of 32 cases of bone marrow specimens from patients initially diagnosed asAML in Hospital of Blood DiseasesAffiliated to ChineseAcademy of Medical Sciences from January 2013 to December 2016, were selected for this study; In addition, AML cell lines (U937, HL60, THP-1, Molm-13) were also collected for this study. The mRNAexpression of GADD45g and E2F1 in above mentioned specimens and cell lines by qPCR,andtheircorrelationwasalsoanalyzed.Thelentiviral vector over-expressing E2F1 (pLV-E2F1-RFP) was constructed to prepare recombinant lentivirus, which was then transfected Molm-13 and THP-1 cells that over-expressing GADD45g. Whether GADD45g exerts tumor inhibition effect on AML cells through inhibition of E2F1 was determined by comet assay, Annexin V/7AAD flow cytometry, β-galactosidase staining and PI staining flow cytometry etc. Results: The mRNA expression of GADD45g was negatively correlated with E2F1 in bone marrow of AML patients and AML cell lines (r=–0.663, P<0.01). Over-expression of GADD45g significantly inhibited the expression of E2F1 in AML cell lines (all P<0.01). Molm-13 and THP-1 cells that simultaneously over-expressing GADD45g and E2F1 were successfully constructed. Compared with the control group, the protein expressions of GADD45g and E2F1 in over-expression groups were significantly increased (all P<0.01). Compared with cells over-expressing GADD45g alone, simultaneous over-expression of both GADD45g and E2F1 significantly reduced the apoptosis, senescence and DNA damage (all P< 0.01), and rescued cell cycle arrest in Molm-13 and THP-1 cells (all P<0.01), thus further reduced the chemo-sensitivity of AML cells caused by GADD45g over-expression (all P<0.01). Conclusion: GADD45g exerts anti-tumor effect inAMLvia inhibition of E2F1.

Down-regulated expression and antioncogenetic role of GADD45g in acute myeloid leukemia / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To quantify the expression of growth arrest and DNA damage inducible protein 45g (GADD45g) gene in the bone marrow samples of patients withAML (acute myeloid leukemia) and inAML cell lines, as well as to study the correlation between the GADD45g expression and prognostic outcome in patients withAML and investigate the role of GADD45g over-expression in proliferation, apoptosis, senescence, differentiation, cell cycle arrest, and drug sensitivity in AML cell lines. Methods: In the study, a total of 27 cases of bone marrow specimens were selected from patients initially diagnosed as AML in Hospital of Blood Diseases affiliated to Chinese Academy of Medical Sciences from January 2013 to December 2016. mRNA and protein expression levels of GADD45g in BMMNCs (Bone marrow mononuclear cells) from patients with AML and healthy donors and in AML cell lines were quantified by quantitative real-time PCR and Western blotting. The correlation between GADD45g expression and overall survival (OS), coupled with event-free survival (EFS) in patients with AML was analyzed in two gene expression datasets (GSE10358, GSE425-GPL317). Lentiviral vectors over-expressing GADD45g were constructed and transfected into AML cell lines (U937, THP-1 and Molm-13 cell lines). The role of GADD45g over-expression in cell proliferation, colony formation, senescence, apoptosis, cell cycle arrest, differentiation and drug sensitivity of U937, THP-1 and Molm-13 cells were detected by cell counting, colony-forming assay, β-galactosidase staining and flow cytometric analysis of Annexin V/7AAD staining, PI staining and so on, respectively. Results: Expression of GADD45g was dramatically down-regulated in BMMNCs in AML patients and AML cell lines compared to that from healthy donors (all P<0.01). The OS (P<0.05) and EFS (P<0.05) of AML patients with low GADD45g expression were significantly shorter that those of AML patients with higher GADD45g level. Enforced expression of GADD45g could inhibit cell growth and colony formation, promote senescence and apoptosis, induce cell cycle arrest and differentiation and enhance drug sensitivity in AML cell lines (P<0.05 or P<0.01). Conclusion: GADD45g expression was remarkably silenced in marrow tissues of patients withAML andAML cell lines; it showed remarkable and all-around inhibiting effect onAMLcell lines, indicating that GADD45g expression has prognostic value inAML.

A Multi-centered Cross-sectional Study of Disease Burden of Pain of Inpatients in Southwest China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Pain is a common burden of disease globally; yet, it is not systematically investigated in China, especially in hospitalized patients. This study was aimed at clarifying the epidemiological characteristics of pain and related factors in hospitalized patients in Southwest China.</p><p><b>METHODS</b>A cross-sectional study was conducted to investigate the prevalence, severity, and influencing factors of pain and modes of postoperative analgesia in hospitalized patients from 17 hospitals in Southwest China. A prevalidated questionnaire was employed to calibrate all of these items within 3 days from March 18, 2015 to March 20, 2015.</p><p><b>RESULTS</b>A total of 2293 patients were surveyed, the incidence of pain was 57.4% in all hospitalized patients at rest, of which 62.1% were with acute pain and 37.9% had persistent to chronic pain. Among surgical patients, 90.8% of them complained of acute postoperative pain at rest and 97.1% in motion. The incidence of acute postoperative moderate-to-severe pain was 28.8% at rest and 45.1% in motion. Surgical patients reported higher incidences of pain, especially acute and persistent pain compared with nonsurgical patients (P < 0.05). Postoperative pain occurred predominately at surgical sites (95.2%) as compared with nonsurgical sites (4.8%). Agedness, lower education level, surgery, and history of smoking were factors associated with increased duration and severity of postoperative pain and nonsurgical pain (P < 0.05).</p><p><b>CONCLUSIONS</b>Pain is a common burden of disease in China, of which surgical pain constituted an important component. Surgical patients complained more severe pain than those who did not undergo surgery. Postoperative analgesia still needs to be improved to control pain after surgery. Patients' perception might influence the efficacy of pain management, which should be implemented with a multidisciplinary approach.</p>

Recent advances and perspective in the study of the molecular imprinting of proteins / 药学学报

Molecular imprinting technique (MIT) involves the synthesis of polymer in the presence of a template to produce complementary binding sites in terms of its size, shape, and functional group orientation. Such kind of polymer possesses specific recognition ability towards its template molecule. Despite the rapid development of MIT over the years, the majority of the template molecules that have been studied are small molecules, while molecular imprinting of proteins remains a significant yet challenging task due to their large size, structural flexibility and complex conformation. In this review, we summarize the research findings over the past five years, and discuss the characteristics of the technique, the most recent progress and the perspective in the field of molecular imprinting of proteins.

Effects of bazheng mixture on urinary amount and contractile-relaxant function of isolated urethral smooth muscle in rabbits / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the pharmacological mechanism of Bazheng Mixture (BZM) in promoting diuresis and relieving stranguria.</p><p><b>METHODS</b>By means of urine collection through indwelling catheter and urethral ring isolation test to observe the effects of BZM on the urinary amount of conscious rabbits and contractile-relaxant function of urethral smooth muscle isolated from rabbits.</p><p><b>RESULTS</b>BZM, at dosage of 5.0 g/kg or 10.0 g/kg by gastrogavage, could increase the urinary amount in 60 min obviously (P < 0.05). For the isolated urethral rings, the maximal contractile and maximal relaxant forces reduced significantly (P < 0.05 or P < 0.01) when the concentration of BZM applied ranged between 9.9-90.9 g/L; the frequency increased significantly (P < 0.05 or P < 0.01) when the concentration ranged 29.1-90.9 g/L and the amplitude increased significantly when the concentration were 56.6 and 90.0 g/L. Along with the concentration of BZM increased from 9.9 g/L to 90.9 g/L, the changes occurred in the above-mentioned parameters of urethral ring were in order of potency amplitude > frequency > maximal relaxant force > maximal contractile force.</p><p><b>CONCLUSION</b>The effects of BZM in enhancing urethral peristalsis may be stronger than that in dilating the urethral caliber. Its mechanism in promoting diuresis and relieving stranguria may be related with its action of urethral dilatation and potential peristalsis promotion.</p>