This thesis aimed at the Bacillus natto TK-1 screened out from Natto.The lipopeptide was purified using Thin-Layer Chromatography(TLC),and investigated its anti-tumor activity.After acid precipitation and methanol extraction,the lipopeptide was separated on TLC.Then the authors get the monomer surfactin which molecular weight is 1036Da through the High performance liquid chromatography(HPLC),electro spray ionization-mass spectrometry(ESI-MS) and infrared(IR).MTT method was implied to testify the anti-tumor activity of the purified sample from TLC.The results indicated a concentration and time-dependent relationships.After 48h,their IC50 were 40 mg/L.The detection with inverted microscope fluorescence microscope displays that the surfactin will cause a series of Morphological changes to the cells.In TUNEL experiment,the authors noticed that surfactin has the ability to induce apoptosis,besides this inhibition shows an obvious time-dependent relationship.

The study was to develop cis-dichlorodiammineplatinum(DDP)-loaded formulations using a novel type of self-assembled compound composed of block copolymers synthesized by poly(?-glutamic acid)(?-PGA).For the potential of targeting liver cancer cells,D-galactose was conjugated on the prepared ?-PGA.In vitro,DDP can be released from the resulting conjugate in PBS:there was a burst release during the first 8 h,then followed by sustained release.DDP could be easily incorporated into poly(?-glutamic acid)-D-galactose esterifiable derivative through a covalent bond.The yield of DDP incorporation into the esterifiable derivative was 9.4%～10.2%.In vitro experiments conclusively established that the poly(?-glutamic acid)-D-galactose esterifiable derivative-Cisplatin Complex Compound(?-D+-DDP)was much less toxic to normal cell lines than DDP only.The surviving rate of cells treated with ?-D+-DDP compound is higher than those treated with free DDP.Also it has obvious antitumor efficiency on human liver tumor BEL-7402 cells.HE staining indicated that the ?-D+-DDP compound make the BEL-7402 apoptosis.These results indicated that the conjugation of DDP to the esterifiable derivative reduced its cytotoxicity activity,but retains its antitumor activity in vitro.In conclusion,the ?-D+-DDP compound could be used as a potential clinic antitumor drug.The ?-PGA obtained by fermentation can be used as a valuable drug carrier system.

DDP could be easily incorporated into poly (?-glutamic acid)-D-galactose esterifiable derivative through a covalent bond. The yield of DDP incorporation into the ?-PGA was 9.4%～10.2%. The DDP was released in the initial 8h in a burst manner,and thereafter in a sustained manner. The results that the conjugation of DDP to poly (?-glutamic acid)-D-galactose esterifiable derivative not only reduced the toxicity of the DDP but also enhanced antitumor activity and the targeting ability. The vivo experiments conclusively established that the ?-D+-DDP compound was much less toxic to animals than DDP alone. A direct evaluation showed that mice treated with ?-D+-DDP compound at a dose of 7.5 mg/kg displayed significant tumor regression. Furthermore,the implanted solid tumors disappeared completely from 35% of the H22 tumor-bearing mice after ?-D+-DDP compound administration. The aforementioned results of biodistributions of the prepared ?-D+-DDP compound in various organs in normal mice demonstrated that the ?-D+-DDP compound had a specific interaction with liver's parenchymal cells and H22 hepatocellular carcinoma tumor cells via ligand receptor recognition. In conclusion,the results indicated that the ?-D+-DDP compound prepared can effectively target the site of hepatoma tumor via the recognition and significantly reduce its size. The ?-D+-DDP compound was less toxic than the free DDP,and could effectively reduce xenografted H22 hepatocellular carcinoma cells in KM mice and prolong the survival of KM mice grafted with H22 hepatocellular carcinoma tumor cells. Therefore,the prepared ?-D+-DDP compound may be used as a potential drug delivery system for the targeted delivery to liver cancers or other liver diseases.

Objective To investigate the effects of different doses of alcohol on the synthesis of testosterone and the expression of androgen binding protein(ABP)mRNA in rat testis.Methods Forty male Wistar rats were randomly divided into 4 groups(10 rats each group)and received either distilled water(control group)or alcohol(alcohol-fed groups)for 5 months.Alcohol was administered by garage with a single daily dose : 5 g/kg(large dose group),2.5 g/kg(middle dose group)and 0.5 g/kg(small dose group).Testosterone content was measured by ELISA.mRNA levels of peripheral-type benzodiazepine receptors(PBR),PPARct and ABP were assayed by RT-PCR.Results Compared with control group:(1)ethanol feeding with daily doses of 5 g/kg,2.5 g/kg and 0.5 g/kg significantly decreased testosterone levels by 31.13%(P0.05)respectively,indicating that ethanol might impair testosterone synthesis;(2) mRNA levels of PBR were decreased in all three ethanol-treated groups(all P

OBJECTIVETo study the effect of ginsenoside on apoptosis of human leukemia-60 (HL-60) cells and its mechanism.

METHODSMTT cytotoxicity assay was used to determine the growth inhibition activity of ginsenoside (100, 50, 25, 12.5, 6.25, 3.125 and 1.5625 μmol/L) on HL-60 cells. The apoptosis of HL-60 cells after treatment with ginsenoside (0,5,10 and 20 μmol/L) was determined by Annexin V-FITC/PI staining and flow cytometry. The cleavage of total proteins by caspase-8, caspase-9 and caspase-3 was evaluated by Western blot. The cleavage of caspase-3 protein was detected by Western blot after treatment with 10 μmol/L ginsenoside and caspase-8 and 9 inhibitors.

RESULTSGinsenoside had potent cytotoxicity on HL-60 cells, with an IC50 value of 7.3±1.2 μmol/L. After treatment with ginsenoside (0, 5, 10 and 20 μmol/L) for 48 hours, the apoptotic rate displayed a dose dependency, as shown by flow cytometry, with significant differences between the groups (F=12.67, P<0.01). Western blot showed that there were caspase-9 and caspase-3 cleavage bands, but without caspase-8 cleavage band. The specific inhibitor of caspase-9 Z-LEHD-FMK could block the caspase-3 cleavage induced by 10 μmol/L ginsenoside, but the specific inhibitor of caspase-8 Z-IETD-FMK did not have this effect.

CONCLUSIONSGinsenoside can induce apoptosis of HL-60 cells, which may be related to a mitochondria-dependent pathway.

Apoptosis ; drug effects ; Caspase 9 ; physiology ; Caspase Inhibitors ; pharmacology ; Ginsenosides ; pharmacology ; HL-60 Cells ; Humans

Objective To explore the experience of developing evidence-based nursing (EBN) course in postgraduate nursing program in order to improve the quality of course.Methods A comprehensive project on developing EBN course for postgraduate nursing students was initiated in School of Nursing Fudan University.59 postgraduate nursing students in school of nursing, Fudan University were conveniently classified into EBN teaching group (n = 33) and control group (n = 26).The students in the EBN teaching group received 54 credit hours evidence-based nursing education.The teaching and learning experience were reflected through faculty interview and student interview. The teaching and learning outcome were evaluated by self-designed questionnaires.Results Students considered EBN course as a challenge.They experienced both positive and negative feelings during learning EBN.The critical appraisal, data extraction, Meta analysis and evidences utilization were seen as bigger challenges. However, they valued the experience of learning EBN as the opportunity for them to integrate knowledge and skill of literature searching, clinical epidemiology, health statistics, and nursing research into the learning of EBN course. Students in EBN teaching group had significantly higher score on evidence-based nursing knowledge than control group (z = 3.582, P < 0.01).Students in EBN teaching group had significantly higher scores on critical appraisal at post-course than at pre-course(t = 3.674,P < 0.01).Most of the students in EBN teaching group were satisfied with course content, teaching materials and teaching methods.In addition, they proposed constructive suggestions on credit hours and teaching arrangement for further implementing of evidence-based nursing course.Conclusions It is suggested that EBN can be developed as a course in postgraduate nursing program.The knowledge and skills on critical appraisal of literature and conducting systematic review can be improved by learning a comprehensive EBN course.However,the course content and teaching methods need further explore.

Objective To evaluate the outcome of the comprehensive evidence-based nursing course on Postgraduate training courses students. Methods Postgraduate training courses students in school of nursing of Fudan University were conveniently assigned to experimental group ( n =22) and control group ( n = 26). The students in the experimental group received 36 hours evidence-based nursing education. The teaching and learning outcome were evaluated by self-designed questionnaires. Results After 36 hours teaching and learning, Students in experimental group got significantly higher score on evidence-based nursing knowledge and literature criticism than control group(Z =3. 582,P<0. 01; t =3. 674,P<0. 01) ; There was no statistical difference on evidence-based nursing attitude score of students in experimental group and control group after evidence-based nursing course(t = 0. 310,P >0. 05); The results of course feedback questionnaire suggested that most of the students in experimental group were satisfied with course content, teaching materials and teaching methods. Interview results showed that students can develop system evaluation and practice Evidence-based nursing. In addition, they proposed constructive suggestions on credit hours and teaching arrangement for further implementing of evidence-based nursing course. Conclusions The knowledge and skill of EBN could be significantly improved by learning a comprehensive evidence-based nursing course for nurses studied in postgraduate nursing Program. Further study is needed to explore the effect of EBN course on students' attitude to evidence-based nursing.

AIM:To explore whether the angiotensin II type 1 receptor autoantibodies(AT1-AA)induces islet β-cell apoptosis and whether autophagy is involved in the process.METHODS:The INS-1 cells treated with AT1-AA at 10-6mol/L for 24 h,and then the apoptosis was analyzed by flow cytometry,Western blot and Hoechst 33258 staining.In addition,the expression of autophagy-related proteins such as LC3 and beclin 1 were determined by Western blot.The effects of AT1-AA on the apoptosis,autophagy and viability of INS-1 cells with or without 3-methyladenine(3-MA;a com-mon autophagy inhibitor)or telmisartan(an angiotensin Ⅱ type 1 receptor blocker)pretreatment, were detected by flow cytometry,Western blot and CCK-8 assay.RESULTS: Treatment with AT1-AA at 10 -6mol/L for 24 h significantly re-duced the cell viability(P<0.05).Compared with the negative IgG control group,the apoptotic cells increased after incu-bation with AT1-AA for 12 h,24 h and 36 h,respectively(P<0.05).Moreover,the protein levels of LC3 and beclin 1 also increased gradually with the prolongation of treatment time,and the elevation of apoptosis and autophagy were blocked by telmisartan.After pretreatment with 3-MA, the apoptotic rate of the cells was obviously decreased compared with the cells treated with AT1-AA alone.CONCLUSION: AT1-AA induces the apoptosis of INS-1 islet βcells by upregulating autophagy via the angiotensin Ⅱtype 1 receptor pathway.

Cystic fibrosis (CF) is a multisystem disease mainly caused by pathogenic mutation of the cystic fibrosis transmembrane conduction regulator gene.In recent years, with the deepened understanding of the disease and the popularization of gene detection technology, an increasing number of children are diagnosed with CF in China.Lung involvement is reported to affect the prognosis of the disease.Lung involvement is closely related to the airway, and good airway management can prolong the life of children.In this paper, the selection of airway clearance techniques and inhaled drugs were expounded, so as to improve the long-term airway management of children with CF in China.

Objective:To investigate the correlation of the clinical characteristics,fever characteristics,serum biomarkers with cytokine release syndrome (CRS) in patients with relapsed/refractory multiple myeloma (R/R MM) treated with chimeric antigen receptor T cell (CAR-T) immunotherapy. Methods:104 R/R MM patients who received CAR-T cell therapy at the Affiliated Hospital of Xuzhou Medical University from June 2017 to November 2021 were included,and the correlations of their clinical characteristics,fever characteristics,serum biomarkers with the severity of CRS were analyzed. Results:Among 104 R/R MM patients receiving CAR-T treatment,no CRS was observed in 8 cases (7.7％),and 96 cases (92.3％) developed CRS. Patients with high-risk cytogenetics had a higher risk of developing CRS (P=0.040),while patients who had previously received autologous hematopoietic stem cell transplantation (ASCT) had a lower risk of developing CRS (P=0.004). There was a significant difference in the duration of fever between patients with grade 1-2 and grade 3-5 CRS (P=0.006). The highest body temperature varied among patients with different treatment regimens (P=0.001). The decrease in total protein in patients with CRS was more significant than in patients without CRS (P=0.002). Within one month after CAR-T cell infusion,the degree of albumin recovery in patients with grade 3-5 CRS was lower than that in patients with grade 0-2 CRS (P=0.037). Compared to patients with grade 1-2 CRS,patients with grade 3-5 CRS showed a significant increase in heart rate after CAR-T cell infusion (P=0.013),while IL-6,C-reactive protein (CRP),and serum ferritin (SF) also showed significant increases (P=0.007,P<0.001,P=0.003). Conclusion:High-risk cytogenetics is a risk factor for severe CRS. Long duration of fever is a clinical characteristic of severe CRS. CRP can better reflect the severity of CRS.