Objective:To clone and express the full length cDNA of human CD38. Methods:The full length cDNA of the human CD38 antigen was amplified from total RNA of Daudi cell by RT-PCR, and it was inserted into pGEM-T. The validity on the sequences was confirmed by automatic DNA sequencing. Inserting the valid CD38 gene into pcDNA3.1(+) plasmid to obtain recombinant mammalian expression vector pcDNA3.1(+)/CD38Z; Using lipofectin gene transfer technique system, recombinant expression vector containing CD38 gene was transfected into COS7 cells. The expression of CD38 molecules on the surface of COS7 cells was detected by FACS and immunohistochemical technique. Results:DNA sequencing showed that the cloned full length cDNA sequence was identical with reported. The result of FACS and immunohistochemical technique indicated that CD38 molecules were expressed on the surface of COS7 cells. Conclusion:The full length cDNA of human CD38 is obtained, recombinant mammalian expression vector pcDNA3.1(+)/CD38Z is successfully constructed, and the CD38 molecules is expressed on the surface of COS7 cells,this may facilitate studies on the biochemistry and function of CD38 antigen.

Objective:To study the effects of ICA on HepG2.2.15 cell proliferation, their sensitivity to the lysis by CD3AK effector cell, to investigate the reversal action of ICA on hepatocarcinoma cells from immune escape through Fas/FasL pathway.To provide the theoretical and experimental bases for ICA development.Methods:MTT assay was used to detect cell proliferation and CD3AK cells cytotoxicity activity;flow cytometry assay was used to examine expression of surface molecules and apoptosis rate of HepG2.2.15 cells.Results:When HepG2.2.15 cells line was treated with 50 ?g/ml ICA,a significant reduction of the rate of cell proliferation was observed. Inhibition rate at 48h was 22.04%,and 29.68% at 72h.Kinetic study showed that inhibition of cell proliferation was time dependent (P0.05).ICA could inhibit apoptosis of Jurkat cells induced by HepG2.2.15 cells. In the co-culture system of HepG2.2.15 cell and Jurkat T cell, apoptosis ratio of Jurkat cell was reduced from 46.66% to 18.20% by ICA (P

Objective To analyze the medication rules of the treatment of stroke in Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan; To provide references for the clinical treatment. Methods Prescriptions with confirmed efficacy of famous TCM doctors in the history in Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan was searched. Excel2003 was used to establish database to analyze medication frequency. SPSS17.0 statistical software was used to conduct cluster analysis, and tree view was used to show results. Results Totally 112 prescriptions for the treatment of stroke of 36 famous TCM doctors were included in the study, including 204 kinds of Chinese materia medica and 1169 times of medication frequency. The high-frequency medicines (>10 times) were Paeoniae Radix Alba, Achyranthis Bidentatae Radix, Angelicae Sinensis Radix, Pinelliae Rhizoma Praeparatum, and Poria. The high-frequency medicine categories were tonifying deficiency medicine (22.58％), pacifying liver and wind medicine (12.31％), activating blood and dispelling stasis medicine (11.89％), clearing heat medicine (11.46％) and dissipating phlegm, cough and asthma medicine (8.72％). Cluster analysis showed that high-frequency medicine (>10 times) could be clustered as 6 categories. Conclusion Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan focuses on tonifying deficiency medicine, accompanied with pacifying liver and wind medicine, activating blood and dispelling stasis medicine, clearing heat medicine and dissipating phlegm, cough and asthma medicine, which can be used to guide clinic.

Objective:To apply the best evidence for prevention and management of enteral nutrition feeding intolerance in patients after upper gastrointestinal surgery to clinical practice and to evaluate its clinical effects.Methods:Guided by the clinical evidence practice application model of the Joanna Briggs Institute (JBI) Evidence-based Healthcare Centre in Australia, the best appropriate evidence was selected based on expert advice and evidence-based practice protocols were developed. The convenient sampling method was adopted to select 84 patients and 21 nurses who were admitted to the Department of General Surgery in a Class Ⅲ Grade A hospital in Taiyuan from September to December 2020 as the research objects. A baseline review of 42 patients and 21 nurses admitted from September to October 2020 was conducted, and education, training and quality supervision were conducted to promote practice change. A total of 42 patients and 21 nurses (same as before) admitted from November and December 2020 were reviewed. Incidence of enteral feeding intolerance, the time to recovery of intestinal function, the knowledge, attitude and practice of nurses towards prevention and management of enteral feeding intolerance and implementation of audit indicators among nurses were compared before and after the application of the program.Results:The incidence of abdominal distension, diarrhea and feeding intolerance in patients after the application of the evidence was lower than that in patients before the application of the evidence, and the difference was statistically significant ( P<0.05) . The first anal exhaust time and first anal defecation time of patients after the application of the evidence were (59.02±17.19) and (68.10±19.98) h, which were shorter than those before the application of evidence (78.79±22.34) and (82.76±22.35) h, and the differences were statistically significant ( P<0.01) . After the application of the evidence, the scores of all dimensions of nurses' knowledge, belief and practice questionnaire for the prevention and management of feeding intolerance were higher than those before the application of evidence, and the differences were statistically significant ( P<0.05) . The implementation rates of the 11 audit indicators were improved from 0-78.57% to 80.95%-100%. Conclusions:The best-evidence based prevention and management strategies of enteral nutrition intolerance in patients after upper gastrointestinal surgery can be applied to clinical practice to standardize the work behavior of nurses in implementing enteral nutrition and reduce the incidence of feeding intolerance in patients.

Objective To analyze the differences in the expressions of the total and phosphorylated proteins in human colon cancer HCT116 cells after the knockout(KO)of retinoic acid-induced protein 16(RAI16)and explore the possible mechanism and related signaling pathways affecting its protein function in HCT116 cells.Methods HCT116 KO and WT cell proteins were collected and extracted,and the protein extraction efficiency was detected via a sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)experiment.After protein digestion,the peptides were labeled with TMT and analyzed via mass spectrometry.We used bioinformatics methods to analyze the identified differential proteins and differentially phosphory-lated proteins by using GO,KEGG,and STRING databases.Results The results of SDS-PAGE showed no evident protein degradation.In addition,some key bands were significantly different between the experi-mental and control groups.A total of 147 up-regulated and 230 down-regulated differential proteins were screened in accordance with the conditions of Foldchange≥1.5 or Foldchange≤1/1.5 and P<0.05.Meanwhile,106 up-regulated and 217 down-regulated phosphorylation sites were screened.GO enrichment analysis revealed that the differential proteins were mainly enriched in the composition of nucleoplasm,nucleus and cytoplasm,RNA binding,cadherin and chromatin,DNA repair,RNA splicing,and positive regulation of DNA as template transcription.The results of KEGG enrichment indicated that the differential proteins were mainly enriched in nucleocytoplasmic transport,spliceosomes,cell cycle,cell-cell tight junctions,viral carcinogenesis,microRNAs in cancer,etc.The protein interaction network mainly focused on DDX17,NCL,EEF2,CDK1,SSRP1,and SMARCC1.The statistical findings unveiled the up-regulated changes in the two omics of SKP1,ORC1,and BAD and the down-regulated changes in RBL1,RB1,CDK1,CDC6,MCM4,TFDP1,CHD4,and SNW1.Moreover,the phosphorylation differences were more significant than the protein differences.Conclusion RAI16 plays the possible crucial role in multiple biological functions and signaling pathways through key proteins,such as SKP1,ORC1,RB1,and CDK1,which affect the cell cycle and thereby the occurrence and development of cancer.

Objective: To investigate whether cancer-associated- fibroblasts (CAF), the key component of tumor microenvironment, regulate the chemoresistant capacity of lung cancer cell line A549 through SDF-1 secretion. Methods: Primary cell isolation techniques was used to isolate cancer-associated-fibroblasts from lung cancer patients. MTT assay was applied to determine the proliferation and chemoresistance of A549 cells. Quantative PCR was used to detect the mRNA changes of Bcl-xL. Western blotting was used to detect the protein expression of Bcl-xL. ELISA was applied to detect the SDF-1 secretion from normal fibroblasts (NF) and CAF. Results: CAF promoted the proliferation of A549 cells, while NF had no significant effect on them. After 72 hrs incubation, the absorbance value of A549+ CAF medium group was 0.814±0.006, significantly different from the 0.753±0.006 of the A549+ NF medium group (P<0.05). The Q-PCR assay indicated that mRNA expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.11, 1.10±0.09 and 3.50±0.30, respectively, showing a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The Western blot showed that protein expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.08, 1.10±0.12 and 3.10±0.25, respectively, with a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The ELISA results showed that the SDF-1 concentrations in the A549+ NF medium group and A549+ CAF medium group were 3.23±0.02 and 9.53±0.10, respectively, significantly different from each other (P<0.05). The MTT assay indicated that the absorbance values of OD of A549 group, A549+ AMD3100 group, A549+ NF medium group, A549+ NF medium+ AMD3100 group, A549+ CAF medium and A549+ CA Fmedium+ AMD3100 group were 0.43±0.03, 0.25±0.02, 0.48±0.03, 0.31±0.03, 0.72±0.06 and 0.45±0.03, respectively. The data of A549+ NF medium group was significantly different from that of A549+ CAF medium group (P<0.05). Conclusions Cancer-associated-fibroblasts enhance the drug resistance of A549 cells through SDF-1 secretion, upregulating the expression level of Bcl-xL through interaction with CXCR4. Our study not only illustrates that tumor microenvironment is able to enhance drug resistance of tumor, but also provides experimental evidence for the cancer-associated-fibroblasts as a potential therapeutic target for the treatment of lung cancer.