Objective To explore the efficient method in detection of DMD/BMD patients.Methods 18 deletion-prone exon fragments of DMD gene were amplified via molecular cloning. They were used as probes and were spotted on the slides treated with APES and poly-lysine together by manual operation to make microarray. In addition, fragments of ?-actin were used as positive contrast and those of pUC 19/EcoR I were used as negative. 30 DMD/BMD patients were detected for deletion in DMD gene with the microarray and 5 healthy people were done as normal control. Parts of the results were compared with PCR method.Results Different exon fragment deletion of DMD/BMD gene was detected in 21 patients by DNA microarray, and 10 of them were confirmed by PCR analysis.Conclusion DNA microarray assay is a convenient ,accurate and sensitive method in diagnosis of DMD/BMD patient.

BACKGROUND: Dystrophin gene is X-linkage recessive heredity nerve-muscle system disease. Dystrophin gene deletions cluster in two hotspot regions, comprising exons 2-20 and 44-53. The majority of deletions can be detected by examining only a subset of exons. However, little is known regarding systematic detection of 18 common deletion exons of dystrophin gene.OBJECTIVE: To obtain and identify the cloning of 18 deletion-prone exons of dystrophin gene.METHODS: A total of 18 fragments of dystrophin gene were obtained through polymerase chain reaction (PCR) amplification with human genomic DNA as template and 18 pairs of primers respectively. The fragments were connected with pGEM-T Easy vector.The recombinants were transformed into E.coli JM109 competent cells, followed by planted on Luria-Bertani (LB)/ampicillin(Amp)/isopropylthio-β-D-galactoside(IPTG)/X-bromo-4-chloro-3-indolyl-β-D-galactoside (X-Gal) plates and cultured.Positive transformants were selected with blue/white color screening, and the recombinant plasmids DNA was extracted and digested with restriction enzyme Not I. DNA sequences of the fragments were analyzed. Nucleotide analyses were performed through the National Center for Biotechnology Information (NCBI) Basic Local Alighment Search Tool (BLAST) against GenBank.RESULTS AND CONCLUSION: Size of the18 fragments by PCR amplification was in accordance with anticipation. Size of the fragments of recombinant cloning by Not I digestion was in accordance with that of PCR and expectation. Sequence size of the 18cloned fragments was in accordance with expectation. The cloned fragments have high homology with dystrophin gene through NCBI BLAST against GenBank. These cloned fragments were the main deletion-prone exons of dystrophin gene.

AIM: To investigate whether viable apoptotic cells and phagocytosis of them affect the activation of T lymphocytes. METHODS: Ultraviolet irradiation was used to induce apoptotic cells in vitro and the model of phagocytosis of these cells was established. Cytokine TGF ?1 was detected by ELISA. The rate of apoptotic cells and phagocytosis of them were assessed by flow cytometry. Furthermore, flow cytometry was also employed to examine the expression of activation signs, such as CD69, CD25, CD71, of T lymphocytes under the intervention of apoptotic cells and macrophage which ingested apoptotic cells, to reflect whether the apoptotic cells and the phagocytosis of these cells could influence the activation of lymphocytes stimulated by Con A. RESULTS: Ingestion of apoptotic cells increased TGF ?1 secretion. Only the macrophages that had ingested apoptotic cells could suppress the activation of lymphocytes. The expression of the markers of lymphocytes activation such as CD69, CD25, CD71 had been restrained. These inhibition effects were abolished by monoclonal anti-TGF ?1 antibody. CONCLUSION: The macrophages that have ingested apoptotic cells inhibit expression of CD69, CD25 and CD71 of T lymphocytes stimulated by ConA. This effect is dependent on the increase in TGF ?1 secretion in local site. [

0.05). However, a significant higher percentage of CD3+CD25+ T cells was found in stable liver transplantation group as compared to healthy group ( P0.05). There was not found no regular change of serum cytokine s (IL-6, TN F-?) and adhesion molecules (ICAM-1, P-selectin) in 6 liver transplanted patien ts during post-operation from day 1 to day 30, indicating that was associated wi th the different status of patients before or after transplantation. CONCLUSIONS: Our data suggesting that increased levels of ICAM-1 and P-selectin, appears to participate in the processing of immunoregulation to transplanted livers, whereas elevated concentrations of IL-6 appear to be involv ed in the repair of the injury induced by TNF-? in allo-transplanted livers.

Objective: To study the effect of different sorts of fertilizer on the content of tanshinone Ⅱ A, the effective element of cultivated Radix Salvia miltiorrhizae. Methods: Fertilize Radix Salvia miltiorrhizae with the several kinds of combination: the oddment cake(round flat cake made of residue of seed after extracting oil form it), pig manure, chicken manure, duck manure, human excrement, plant ashes, dregs of a decoction (residue of Traditional Chinese medicine material after being extracted), phosphoric fertilizer and compound fertilizer; and gather Radix Salvia miltiorrhizae one year later. Determine the content of Tanshinone Ⅱ A in cultivated Radix Salvia miltiorrhizae by HPLC. Results: The contents of Tanshinone Ⅱ A in Radix Salvia miltiorrhizae fertilized by different sorts of fertilizer are of significant difference. Conclusion: Plant ashes, oddment cake and compound fertilizer are good for growing of Radix Salvia miltiorrhizae and raises of the content of Tanshinone Ⅱ A.

Objective To study the effect of interleukin (IL)-10 knockout (IL-10-/-) on renal repair after renal ischemia-reperfusion injury in mice.Methods Eighteen IL-10-/-mice (KO) aged 8-10 weeks and 18 C57BL/6 wild type mice (WT) aged 8-10 weeks were divided into control group (Sham) and renal ischemia-reperfusion injury (IRI) group.The renal tissue morphology change was observed by Hematoxylin and eosin (HE) staining and Masson staining.The expressions of IL-18, Ki67 and TGF-β1 were detected by immunohistochemistry.The expression of TGF-beta1 and IL-18 were detected by Western blotting.Results Compared with that in WT-IRI group, in KO-IRI group renal pathological damage was more severe, renal interstitial fibrosis was visible, Ki67 expression of renal tubular epithelial cells decreased distinctly (P＜0.01), the expression of TGF-betal increased significantly (P＜0.01).Conclusion Repair slows down significantly after kidney ischemia-reperfusion injury and fibrosis occurs gradually in IL-10-/-mice, eventually progressing to chronic kidney disease.

Background Self-perceived burden,anxiety and depression are among the most important factors affecting quality of life.At present,there is a lack of understanding on the research status and influencing factors of self-perceived burden in liver transplant recipients.Previous studies have shown that self-perceived burden,anxiety,depression and quality of life are correlated in pairs,but the effecting path among the three are not yet clear.Objective To explore the correlation of self-perceived burden and anxiety/depression with quality of life in liver transplant recipients,so as to provide guidance for psychological nursing intervention in clinical patients.Methods A total of 200 patients liver transplant recipients were enrolled from the liver transplantation inpatient and outpatient clinics of Jiangsu Province Hospital and Qinhuai Medical Area,General Hosptial of Eastern Theater Command of People's Liberation Army of China from March 2022 to February 2023.Patients were evaluated using Self-perceived Burden Scale(SPBS),Hospital Anxiety and Depression Scale(HADS)and the Chinese version of Post Liver Transplant Quality of Life Questionnaire(pLTQ).Spearman correlation analysis was used to examine the correlation among the scales.A structural equation model using Mplus 8.3 was utilized to testify the relationship among self-perceived burden,anxiety/depression and quality of life in liver transplant recipients.Bootstrap method was used to test the effecting pathway.Results There were statistically significant differences in SPBS scores of liver transplant recipients with different levels of education and fannual family income(H=9.656,18.796,P<0.05).There were statistically significant differences in HADS scores of liver transplant recipients with different numbers of somatic symptoms(H=9.859,P<0.05).There were statistically significant differences in the Chinese version of pLTQ scores of liver transplant recipients with different levels of education,postoperative survival time and numbers of somatic symptoms(H=6.892,8.023,16.099,P<0.05).The total and each dimension scores in SPBS of liver transplant recipients were positively correlated with the total score and anxiety/depression dimension scores in HADS(r=0.464～0.586,0.460～0.593,0.286～0.408,0.464～0.583,P<0.01)and negatively correlated with the total score and each dimension scores in the Chinese version of pLTQ(r=-0.572～-0.416,-0.599～-0.441,-0.365～-0.213,-0.559～-0.428,P<0.01).Structural equation model denoted that self-perceived burden negatively affected quality of life(β=-0.186,P<0.01).Anxiety/depression also negatively affected quality of life(β=-0.679,P<0.01).The self-perceived burden indirectly affected the quality of life of liver transplant recipients through anxiety and depression,with an effect value of-0.429,accounting for 69.76％of the total effect.Conclusion The quality of life in liver transplant recipients may be related to their self-perceived burden and anxiety/depression.Self-perceived burden may affect the quality of life of liver transplant patients through anxiety and depression.

Objective To develop an expert consensus on subcutaneous injection for allergen-specific immunotherapy.Methods Relevant domestic and intemational literature was reviewed,and nursing experts who had experiences in subcutaneous injection of allergen-specific immunotherapy were interviewed to form the initial draft of the consensus.A total of 85 experts from 42 hospitals nationwide were invited to participate in discussions.2 rounds of expert consultations,adjustments,revisions,and improvements were made to the initial draft,and an online meeting was held to form the final version of the consensus.The content approved by more than 75％of the expert group is adopted,or it will be discussed or deleted.Results The expert consensus includes operational standards for subcutaneous injection of allergen-specific immunotherapy,identification and management of adverse reactions,and health education.Conclusion The consensus demonstrates strong scientific rigor and practicality,providing guidance for nursing practices in the field of clinical allergology.