Objective:This study aims to analyze the differences in the alternative splicing pattern of ADAR2 among glioma cell lines U87, U251, A172, and normal human astrocyte HA1800. Methods:A-to-I editing level at the Q/R-Site of GluR-2 was analyzed by RT-PCR and sequencing. Real-time PCR was performed to detect the expression level of each alternatively splicing variant using a specific primer that was confirmed to amplify only the targeted template and not other alternatively spliced variant fragments. Results:We verified that the Q/R-Site of GluR-2 is under-edited in glioma cell lines. Real-time PCR revealed that the ADAR2 pre-mRNA splic-ing pattern has no significant difference at exons 1a and 2 between glioma cell lines and normal human astrocyte. We also detected that the amount of alternative splicing variants, including exon 5a, was higher than that of alternative splicing variants not including exon 5a in human glioma cell lines. However, the expression of alternative splicing variants, including exon 5a, was lower than that of alterna-tive splicing variants not including exon 5a in human astrocyte. Conclusion:Evident differences in splicing were observed at the site of exon 5a between glioma cell lines and normal human astrocytes. The difference in the alternatively splicing pattern at exon 5a may be attributed to the decreased activity of ADAR2.

Objective: To observe the clinical efficacy of grain-sized moxibustion in treating chemotherapy-induced myelosuppression for non-small cell lung cancer (NSCLC) and its effect on quality of life (QOL). Methods: Eighty NSCLC patients admitted to the Inpatient Department of Zhejiang Cancer Hospital between September 2016 and March 2018 were recruited and divided into an observation group and a control group by random number method, with 40 cases in each group. The two groups both received chemotherapy with paclitaxel plus cisplatin (TP regimen). The control group received oral administration of leucogen tablets starting from the first day of chemotherapy, 20 mg each time, three times a day, for consecutive 14 d; the observation group was additionally given grain-sized moxibustion, once a day, five days per week at a two-day interval, until the fourteenth day. The myelosuppression severity was observed and compared between the two groups prior to chemotherapy, at the 3rd, 7th and 14th days of chemotherapy; the QOL in the two groups was evaluated before chemotherapy, at the 14th and 21st days of chemotherapy. Results: Regarding myelosuppression, the peripheral blood indicators increased significantly at the 3rd day of chemotherapy in both groups (P<0.05 or P<0.01); at the 7th and 14th days of chemotherapy, the peripheral blood indicators presented a decreasing tendency in the two groups, but the level in the observation group was still significantly higher than that before chemotherapy (P<0.01); at the 3rd, 7th and 14th days of chemotherapy, the peripheral blood indicators in the observation group were higher than those in the control group (P<0.05 or P<0.01); the occurrence rate of myelosuppression in the observation group was significantly lower than that in the control group (P<0.01). The QOL score in the observation group was markedly higher than that in the control group at the 14th and 21st days of chemotherapy (both P<0.05). Conclusion: Grain-sized moxibustion can effectively improve myelosuppression after chemotherapy for NSCLC, reducing its occurrence and enhancing the patient's QOL.

Objective:To investigate the diagnostic value of conventional endoscopy (CE) and endoscopic ultrasonography (EUS) for invasion depth prediction of superficial gastric cancer.Methods:A total of 84 patients with superficial gastric cancer underwent both CE and EUS before treatment at Beijing Shijitan Hospital from January 2011 to December 2019. The patients were divided into CE affirmation group (47 cases) and CE non-affirmation group (37 cases) according to the endoscopist′s affirmation in the results of CE. Diagnostic accuracy of each method was compared with the histology of the resected specimen. And influential factors for the diagnosis were analyzed.Results:The overall accuracy in determining the invasion depth of superficial gastric cancer was 73.8% (62/84) for CE and 81.0% (68/84) for EUS respectively ( P=0.092). In CE affirmation group, the diagnostic accuracy of CE was significantly higher than that in the CE non-affirmation group [93.6% (44/47) VS 48.7% (18/37), χ2=21.656, P<0.001]. Twenty (23.8%) of 84 lesions were over-staged by CE, dignosed as surgical candidates, and 8 (40.0%) of the over-staged diagnosis were modified by additional EUS assessment. Multivariate logistic analysis showed that influential factors associated with observer affirmation included uneven surface of lesion ( OR=5.076, 95% CI: 1.628-15.821, P=0.005), margin elevation ( OR=3.831, 95% CI: 1.238-11.857, P=0.020) and undifferentiated carcinoma ( OR=6.887, 95% CI: 1.882-25.204, P=0.004). Conclusion:For patients of CE affirmation in the invasion depth, the diagnostic accuracy is high. For those of non-affirmation, additional EUS can improve the diagnostic accuracy and help to develop a more appropriate regime.

This study was purposed to explore the effect of specific small interfering RNA targeting at bcr-abl fusion gene and its combination with p27 gene clone on the proliferation, cell cycle and apoptosis of chronic myeloid leukemia (CML) cell line K562. CML cell line K562 was used as the study object. A 21nt siRNA targeting at the fusion site of b3:a2 mRNA in bcr-abl fusion gene was designed, synthesized and transfected into the K562 cells as RNA interference group. Northern blot was used to detect the bcr-abl fusion gene, Western blot was used to detect the expression of P210 protein and apoptosis-related protein BCL-xL after the transfection. Meanwhile, p27 gene was amplified from peripheral blood mononuclear cells by RT-PCR, and was confirmed to be correct by sequencing, then p27-pcDNA3.1 vector was constructed and transfected into K562 cell line by Lipofectin. After being selected with G418, p27-pcDNA3.1-K562 cell clone stably expressing p27 was isolated. P27 protein was identified by Western blot. Finally, siRNA and p27 gene clone were together applied to K562 cells, the cell survival rate was tested by MTT. The cell cycle and the apoptosis were tested by flow cytometry. The result showed that in contrast with the control group, the expression level of bcr-abl fusion gene was much lower in siRNA group, about 18.4% of K562 cells in siRNA group were apoptotic at 24 hours after siRNA transfection, and the expression of apoptosis-associated protein BCL-xL was greatly down-regulated. The expression of P27 protein could be detected by Western blot in p27-pcDNA3.1-K562 cells. The strong inhibition of cell proliferation was observed in p27-pcDNA3.1-K562 cells, as compared with control K562 cells. The count of p27-pcDNA3.1-K562 cells in G(0)/G(1) phase increased apparently, but that in S phase declined greatly. Cell cycle was arrested in G(0)/G(1) phase. After the combination of p27-pcDNA3.1-K562 cells with specific siRNA, the percentage of apoptosis obviously increased and cell survival rate significantly declined. It is concluded that the specific siRNA distinctly inhibits the expression of bcr-abl fusion gene, and can induce K562 cell apoptosis. The combination of specific siRNA with P27 gene clone displays a synergy of inhibition and pro-apoptosis effects to K562 cells.
Apoptosis ; Cell Cycle ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; Fusion Proteins, bcr-abl ; genetics ; Humans ; K562 Cells ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

Graft Rejection ; Humans ; Liver Transplantation

The aim of the manuscript was to optimize formulations and preparation technologies of cataplasm of white mustard seed varnish, and to evaluate its anti-asthma effect on rats. The single factor experiments included spreading thickness, types of crosslinking agents, dihydroxyaluminum aminoacetate amount, sodium polyacrylate amount, types of adhesive agents with human sense as the evaluation index. Blank cataplasm matrix was optimized by the orthogonal experiment with the amount of glycerine, citric acid, and sodium carboxymethylcellulose as the major influential factors. Initial adhesive force, peeling strength and human sense were as the evaluation index. The optimized formulation of blank cataplasm were as followings: glycerine-water-ethanol-PEG400-dihydroxyaluminum aminoacetate-citric acid-sodium carboxymethylcellulose-sodium carboxymethylcellulose 2 : 8 : 0.8 : 0.4 : 0.07: 0.15 : 0.1 : 0.5. The active ingredients of white mustard seed, corydalis, and gansui root were extracted by alcohol extraction method. Asiasarum volatile oil was extracted by oil extractor. The optimized drug loading amount was 11% with initial adhesive force, peeling strength and human sense as the evaluation index. Asthma rats model were established by sensitized with ovalbumin and nose-scratching time as the evaluation index. High dose (17%) group of drug-loaded cataplasm had the obvious inhibition effect on nose-scratching time of rats (P = 0.037 < 0.05). In comparison, middle dose (11%), low dose (4%) and positive-control groups had no obvious inhibitive effect on rats. White mustard seed cataplasm supplied a novel choice for anti-asthma therapy. And the overall pharmacodynamics assessment will be carried out on molecular level in near future.
Animals ; Anti-Asthmatic Agents ; administration & dosage ; chemistry ; Asthma ; drug therapy ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Humans ; Male ; Mustard Plant ; chemistry ; Rats ; Rats, Sprague-Dawley ; Seeds ; chemistry

In recent years, layer-by-layer self-assembly (LbL) has developed rapidly. It has been widely used in various industries such as medicine and metallurgy because of its simplicity, flexibility and controllability. In the study of drug delivery system, hollow microcapsules constructed by LbL method as drug carrier have great advantages in drug release, circulation in vivo and bioavailability, providing a technical platform for targeted drug release. In this paper, we summarize the types of film-forming materials and the driving force used in LbL technology, the way of loading drug into hollow micro capsule, and the variety of loaded drugs. We focus on the release mechanism, its evaluation and safety evaluation of self-assembled film as drug carrier in vivo and in vitro. The review shows the great application prospect of LbL technology in the field of drug delivery.

The Streptococcus suis serotype 2 (S. suis 2) isolates 05ZYH33 and 98HAH33 have caused severe human infections in China. Using a strand-specific RNA-seq analysis, we compared the in vitro transcriptomes of these two Chinese isolates with that of a reference strain (P1/7). In the 89K genomic island that is specific to these Chinese isolates, a toxin–antitoxin system showed relatively high levels of transcription among the S. suis. The known virulence factors with high transcriptional activity in these two highly-pathogenic strains are mainly involved in adhesion, biofilm formation, hemolysis and the synthesis and transport of the outer membrane protein. Furthermore, our analysis of novel transcripts identified over 50 protein-coding genes with one of them encoding a toxin protein. We also predicted over 30 small RNAs (sRNAs) in each strain, and most of them are involved in riboswitches. We found that six sRNA candidates that are related to bacterial virulence, including cspA and rli38, are specific to Chinese isolates. These results provide insight into the factors responsible for the difference in virulence among the different S. suis 2 isolates.

BACKGROUND:The formation of postoperative abdominal adhesions is a complicated process,and the prevention of postoperative adhesions is an urgent problem in clinic. OBJECTIVE:To analyze the mechanism of adhesion at cellular and molecular levels,and to provide theoretical basis for the prevention and treatment of adhesion by mesenchymal stem cell exosomes. METHODS:"Abdominal adhesion,pelvic adhesion,postoperative adhesion,epithelial mesenchymal transformation,mesenchymal stem cells,stem cell exosomes,mesenchymal stem cell exosomes"were selected as Chinese and English search terms.We searched PubMed,CNKI,and Chinese biomedical literature and screened relevant articles on postoperative abdominal adhesion and mesenchymal stem cell exosomal intervention published from inception to August 2023.After systematic analysis,54 articles were finally included for the review. RESULTS AND CONCLUSION:(1)Any pathological factors such as peritoneal inflammation,mechanical injury,tissue ischemia,and foreign body implantation cause peritoneal surface injury,resulting in postoperative abdominal adhesion.The formation process of adhesion includes the interaction of peritoneal mesothelial cell repair,inflammatory response,fibrinolytic system,coagulation pathway and other processes,involving a variety of cytokines and signaling pathways.Wnt/β-catenin pathway can induce fibrosis and angiogenesis,and cooperate with transforming growth factor-β/Smads signaling pathway to stimulate fibroblast proliferation and cause peritoneal fibrosis.Meanwhile,nuclear factor-κB signaling pathway up-regulates the expression of cellular inflammatory factors,promotes fibroblast proliferation,and plays a key role in the process of tissue fibrosis.(2)The paracrine function of stem cells is an important direction of molecular intervention in abdominal adhesions based on regenerative medicine.It can participate in a variety of complex cytokines and signaling pathways involved in abdominal adhesions.(3)Compared with traditional methods for treating abdominal adhesions,mesenchymal stem cell exosome has biological activity and is safe to use.Mesenchymal stem cell exosomes without special culture and expansion have lower immunogenicity,longer stability and other advantages,can guide a normal repair and healing through a variety of ways.(4)Mesenchymal stem cell exosome has been proven to be involved in regulating the above processes of adhesion formation in previous studies,showing potential application prospects in clinical studies.However,further clinical studies are needed to explore appropriate treatment options for mesenchymal stem cell exosomes to address the problem of clinical translation.

The Bacillus subtilis Sac B gene with the vacuolar targeting signal sequence driven by 35S promotor was transferred into Lespedeza thunbergii by Agrobacterium mediated method. Total 62 Kan-resistant plants were obtained, of which 5 plants were proved to be transgenic plants. The transgenic plants were characterized by PCR amplification, PCR-Southern hybridization and RT-PCR. The physiological assay results showed that the transgenic plants were more tolerant to stress than the controls under the condition of 200mmol/L NaCl and 5% PEG, respectively, and that the content of soluble sugar in trnsgenic plants was significantly higher than that of controls in the period of tests (5-15 days) under salt and PEG stress.
Bacillus subtilis ; enzymology ; genetics ; Carbohydrate Metabolism ; Carbohydrates ; chemistry ; Hexosyltransferases ; genetics ; Lespedeza ; drug effects ; genetics ; growth & development ; metabolism ; Plants, Genetically Modified ; Reverse Transcriptase Polymerase Chain Reaction ; Sodium Chloride ; pharmacology ; Solubility ; Stress, Physiological ; drug effects ; Transformation, Genetic ; Transgenes ; genetics