Objective: To investigate the best extracting conditions of Polygonatum sibiricum polysaccharides by single and compound enzymatic methods, and to study on the change content of polysaccharide in processing of P. sibiricum by steamed and shined for nine times, which provide theoretical basis for processing of P. sibiricum and other Chinese herbal medicines. Methods: Response surface method (RSM) was used to optimize the extraction of polysaccharides in P. sibiricum. PMP precolumn derivatization and HPLC characteristic map technology were used to identify and analyze the changes of polysaccharide of P. sibiricum being steamed and shined for nine times and establish polysaccharide characteristic map of P. sibiricum by ancient processing and distillate. The effect of different enzymes on the extraction efficacy of P. sibiricum polysaccharides was also compared. Results: The extracting effect of cellulase in single enzyme was the best. Moreover, cellulose and pectinase were also the most significant factor for extracting polysaccharides from P. sibiricum in four complex enzymes. The best extracting conditions for polysaccharide obtaining by RSM as follows: Cellulase temperature was 51 ℃, cellulase dosage was 2%, and the total polysaccharide content was (675.34 ± 0.10) mg/g when the pH value of enzymolysis was 5.40. PMP-HPLC was used to detect four processed P. sibiricum, which contained mannose, ribose, rhamnose, glucose, and galactose. Glucose had a high proportion in P. odoratum, P. cyrtonema, and P. sibiricum polysaccharide, and P. filipes polysaccharide mainly composed of mannose. With the increase of processed times, the content of each monosaccharide increased first, then decreased, then increased and then tended to be stable. The content of mannose and galactose varied greatly. The content of monosaccharides in the distillate increased first and then stabilized, and the content changes of mannose and glucose were obvious. Conclusion: The extraction efficiency and purity of P. sibiricum polysaccharides can be improved by single and compound enzymatic extraction with cellulase, and the extraction yield of polysaccharide in P. sibiricum could be affected by steamed and shined for nine times. PMP-HPLC characteristic map technology can be used to determine the content of the polysaccharide in processing of P. sibiricum accurately and reproducibly, which can be used for the quality evaluation of P. sibiricum and other Chinese herbal medicines.

Objective:To study the application value of WeChat video teaching mode in clinical teaching of digestive endoscopy nurses.Methods:The study selected 126 nursing trainees taking an internship in the Department of Gastroenterology in our hospital from January 2018 to June 2020 as the teaching objects, and they were randomized into two groups, Control Group and Experimental group, with 63 cases in each group. The control group adopted traditional teaching and the experimental group took traditional teaching combined with WeChat video teaching. The internship time of both groups was 4 weeks. The scores of comprehensive ability assessment after practice were compared between the two groups, and the critical thinking ability scale and nursing students' clinical practice satisfaction scale were used to evaluate the critical thinking ability and satisfaction of practice nurses of the two groups before and after teaching. SPSS 19.0 was used for statistical analysis of the research data. Continuous variables were expressed as (mean ± standard deviation), t test was performed, categorical variables were expressed as percentage, four-grid test was conducted, and the test level was α=0.05. Results:The scores of comprehensive ability, theoretical knowledge and operation skills of the experimental group were significantly higher than those of the control group ( P<0.05). After teaching, the scores of seeking truth, opening mind, systematization ability, analytical ability, curiosity, cognitive maturity and total scale score of the two groups were significantly higher than those before teaching ( P<0.05), and there was no significant difference in the score of thinking self-confidence before and after the teaching ( P>0.05). The scores of seeking truth, opening mind, systematization ability, analytical ability, curiosity, cognitive maturity and total scale score of the experimental group were significantly higher than those of the control group ( P<0.05), and there was no significant difference in the score of thinking self-confidence between the two groups ( P>0.05). The experimental group's practice management, practice environment, teaching content, practice effect score and total scale score were significantly higher than those of the control group ( P<0.05), and compared with the control group, the score of teaching quality was poorer, without significant differences ( P>0.05). Conclusion:The application of WeChat video teaching mode in clinical teaching of digestive endoscopy nurses can improve the performance and satisfaction of comprehensive ability assessment of practice nurses, and enhance their ability of critical thinking.

Objective To investigate the relationship between refractory hypertension and renal hemodynamics in end stage renal diseases (ESRD) patients.Methods ESRD patients were classified into:patients with refractory hypertension (group A) and patients with normal blood pressure(group B).Renal hemodynamic indices were ex- amined by duplex ultrasonography.Fasting serum lipid (TC,TG,HDL-C,LDL-C,Lp(a),ox-LDL) and serum parathyroid hormane (PTH) were determined in all patients.Results Significant differences were found in renal hemodynamic indices such as peak systolic velocity (PSV),mean flow velocity (MV),pulsatility index (PI),renal- aortic ratio (RAR) and in clinical index such as Lp(a) and ox-LDL between the two group.Refractory hyperten- sion patients had lower renal hemodynamic indices and higher Lp(a) and ox-LDL levels than in patients with con- trolled BP.Logistic regression analysis revealed that refractory hypertension was related with PSV,EDV,Pl and RAR,but not relevant with sex,age,dialysis time,hematocrit,BUN,creatinine,TC,TG,HDL-C,LDL-C, PTH,MV and RI.Conclusion Atherosclerotic renal artery stenosis and severe disorder in renal hemodynamics is likely the cause for refractory hypertention in ESRD patients.The rise of serum Lp(a) and ox-LDL might acceler- ate renal artery atherosclerosis.

Objective To explore the relationship of vascular endothelial growth factor(VEGF),IFN-? and IL-4 and effect of Bude-sonide on their in asthmatic rats.Methods Thirty-six male SD rats were randomly divided into 3 groups:asthma group,Budesonide-treatment group and control group.On the first day of the experiment and the 8th day,the rat models of the asthma group and Budesonid treatment group were allergized by the OVA/Al(OH)3 through intraperitoneal injection,respectively.And starting from the 15th day,they were challenged by the OVA through atomization for 2 weeks.Control group was allergized and challenged by NS atomization.Budesonid treatment group was interfered in Budesonide inhalation before suscitation in 0.5 h.After 12 h the same inhal done was again in Budesonide group.Twenty-four hours after the last challenge,the rats in 3 groups were sacrificed,and blood and bronchoalveolar lavage fluid(BALF) were collected.The concentrations of IL-4,IFN-? and VEGF in serum and BALF were measured by enzyme-linked immunosorbent assay.Results The concentrations of IL-4 in serum and BALF in asthma group and Budesonide treatment group were significantly increased than those in control group(P

OBJECTIVETo investigate whether myocardial bridging (MB) is independently associated with coronary atherosclerosis proximal to MB in the left anterior descending coronary artery (LAD) identified by computed tomographic coronary angiography (CCTA).

METHODSFrom March 2011 to December 2012, patients (n=9 862) with suspected coronary disease underwent CCTA using dual-source CT scanner. The baseline clinical characteristics (age, gender, smoking history, presence of hypertension, dyslipidemia, diabetes mellitus, family history of heart attack, and body mass index) and the results of CCTA were reviewed. Two radiologists evaluated the coronary artery for MB and coronary atherosclerosis stenosis (CAS) in LAD and made a diagnosis by consensus. Significant independent risk factors for CAS were investigated by multivariate logistic regression analysis.

RESULTSA total of 3 182 (32.3%) cases of MB and 3 359 cases of CAS of LAD were identified. No patient with CAS in the tunneled segment was found. The mean length of bridges and the mean thickness of the overlying myocardium was (17.3±5.2) mm and (1.2±0.9) mm, respectively. There were 1658 MB cases in 3 359 cases of LAD stenosis and 1 524 MB cases in 6 503 cases of no LAD stenosis (χ(2)=681.12, P<0.05). Logistic regression analysis showed that MB in the LAD were significantly associated with CAS in the proximal LAD (OR=3.07, 95%CI=2.81-3.37, P<0.001), and after final adjustment for age, gender, body mass index, family history of heart attack, smoking, hypertension, dyslipidemia, diabetes mellitus, and resting heart rate (OR=2.86, 95% CI=2.60-3.16, P < 0.001).

CONCLUSIONMB in the LAD is independently associated with CAS in the proximal segment to MB.

Adult ; Aged ; Aged, 80 and over ; Coronary Stenosis ; complications ; diagnostic imaging ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Myocardial Bridging ; complications ; diagnostic imaging ; Risk Factors ; Tomography, X-Ray Computed

Background and Objective:The mechanism of tumor tissues selectively uptaking the photosensitizer in photodynamic therapy(PDT)is still unclear.This study was to investigate the affinity of tumor cells to the photosensitizer photofrin-Ⅱ. Methods: Ultraviolet spectrophotometer was applied to measure the absorption spectra of various cell culture media.The fluorescence spectrum of photofrin-Ⅱ was determined by spectrofluorometer.The absorption and eIimination condition of photofrin-Ⅱ were detected in immortalized human esophageal epithelial cell line SHEE and its malignant transformation cell line SHEEC.Results:The maximum excitation wavelength of fluorescence for photofrin-Ⅱ was(395.0±0.5) nm, and the maximum emission wavelength of that was (634.1±0.5) nm.The laser at the wavelength of 630 nm used in this experiment could permeate various types of cell culture media.There was no significant difference in the absorption and elimination of photofrin-Ⅱ between SHEE and SHEEC at the same concentration and time.The absorption of photofrin-Ⅱ in SHEE and SHEEC increased with the increase in photofrin-Ⅱ concentration and duration. and reached the platform at the concentration of 30 μg/mL and a time point of 150 min.respectively.The photofrin-Ⅱ contents of SHEE and SHEEC showed a slight change after 15-30 min, and diminished rapidly after 30 min.Conclusion:High photosensitizer concentration in tumor tissues may be not correlated with the affinity of tumor cells.

Objective To establish a quantitative RT-PCR method with self-quenched fluorogenic probe for detection of bcr/abl mRNA in patients with chronic myeloid leukemia for providing a useful tool for diagnosis of CML,evaluation of therapeutic effect and monitoring of minimal residual disease(MRD). Methods bcr/abl gene from cultured K562 cells was amplified by conventional RT-PCR.The standard quantitative plasmid was constructed by A-T clone method.The self-quenched fluorogenic quantitative RT- PCR method(FQ-RT-PCR)for determination of bcr/abl mRNA was established successfully using the ABI PRISM 7000 PCR Detector.The linear range,sensitivity,stability,and repetitiveness of the method were determined.The marrow samples from 25 CML patients and 3 ALL patients were assessed.Results The sensitivity of the FQ-RT-PCR was 10 copies/?l recombined plasmid,and bcr/abl mRNA can be detected from 1 K562 cell in 10~5 normal cells.The linear range was 10~2-10~9 copies/?l recombined plasmid.The coefficient variation(CV)value was 2.1% in intra-assay and 6.1% in inter-assay.The median ber/abl mRNA expression level was 4.50?10~4 copies/?g RNA [(0.45-89.00)?10~4],5.45?10~4 copies/?g RNA [(2.95-19.30)?10~4 ],13.00?10~4 copies/?g RNA [(4.10-89.00)?10~4] and 2.35?10~4 copies/?g RNA [(0.45-5.12)?10~4] in 25 CML patients,11 patients in the incipient chronic phase,6 patients in blastic crisis,8 patients in chronic period after treatment,respectively.The bcr/abl mRNA level in blastic crisis was significantly higher than that in chronic phase(q= 3.41,P

OBJECTIVESTo investigate targeted blockage of BCR/ABL oncoprotein mediated cell transformation by STAT5 decoy oligodeoxynucleotide (ODN), its effect on the growth and proliferation inhibition of K562 cells and the related molecular mechanisms.

METHODSSTAT5 decoy ODN, designed and synthesized in vitro, was transfected into K562 cells by cationic lipid. The cell growth curve and colony formation assay were used to reflect the growth and proliferation capacity of K562 cells, RT-PCR to detect the expression of three genes downstream STAT5.

RESULTSConfocal microscopy demonstrated that STAT5 decoy ODN was successfully transfected into K562 cells (95.2% positive cells). STAT5 decoy ODN inhibited the growth of K562 cells (inhibition rate 77.7%) and their colony formation capacity (Decoy ODN treated group 8.3% vs control group 35.7%, P < 0.05) after the treatment with STAT5 decoy ODN, the expressions of c-myc, bcl-X(L), cyclin D1 mRNA were down-regulated by 15.4%, 30.8%, 29.1%, respectively in the K562 cells.

CONCLUSIONSSTAT5 decoy ODN inhibits the growth and proliferation of K562 cells. The mechanisms may be that decoy ODN blocks the transcriptional activation potent of STAT5 and down-regulates the expression of these tumor related genes downstream STAT5.

Cell Proliferation ; Cyclin D1 ; genetics ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Gene Expression ; Humans ; K562 Cells ; Liposomes ; Microscopy, Confocal ; Oligodeoxyribonucleotides, Antisense ; genetics ; Proto-Oncogene Proteins c-myc ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; STAT5 Transcription Factor ; genetics ; physiology ; Transfection ; bcl-X Protein ; genetics

OBJECTIVETo evaluate the value of normalization window of tumor vasculature (NWTV) in patients with unresectable gastric cancer undergoing neoadjuvant chemotherapy.

METHODSFrom October 2010 to March 2011, 93 patients with unresectable advanced or locally advanced gastric carcinoma were prospectively collected and randomly divided to Group A(n=30), Group B(n=29), and Group C(n=34). Group A received FOLFOX4 as conventional neoadjuvant chemotherapy. Group B received FOLFOX4 plus bevacizumab. The treatment was adjusted in Group C according to the hypothesis of NWTV with neoadjuvant chemotherapy delivered 5 days after bevacizumab treatment. The efficacy, drug toxicity and clinical outcome were assessed and compared between the three groups.

RESULTSThere were no significant differences among the 3 groups in demographics(P>0.05). All the patients completed the neoadjuvant chemotherapy. Efficacy and toxicity between the three groups were comparable(P>0.05). The rates of tumor downstaging in the three groups were 56.7%(17/30), 72.4%(21/29), 85.3%(29/34), respectively, with a significantly lower downstaging rate in Group C as compared to Group A(P<0.05). R0 resection rates were 23.3%(7/30), 27.6%(8/29), 52.9% (18/34), respectively, with significantly higher R0 resection rate in Group C as compared to Group A and Group B(All P<0.05). There was no perioperative death in this cohort. Postoperative complications were comparable among the 3 groups(P>0.05).

CONCLUSIONSAnti-angiogenesis agent can improve the efficacy of neoadjuvant chemotherapy in unresectable gastric cancer. Furthermore, administration according to NWTV may achieve better outcomes.

Adolescent ; Adult ; Aged ; Antibodies, Monoclonal, Humanized ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bevacizumab ; Female ; Fluorouracil ; therapeutic use ; Humans ; Leucovorin ; therapeutic use ; Male ; Middle Aged ; Neoadjuvant Therapy ; Neovascularization, Pathologic ; drug therapy ; Organoplatinum Compounds ; therapeutic use ; Prospective Studies ; Stomach Neoplasms ; blood supply ; drug therapy ; Treatment Outcome ; Young Adult

OBJECTIVETo observe the inhibitory effect of 4-chlorobenzoyl berbamine (BBD9) on imatinib-resistant cell line K562 (K562/IR) in vitro and in vivo and explore the mechanisms.

METHODSThe IC50 of BBD9 and berbamine (BBM) was determined by MTT assay. The expressions of p210(Bcr-Abl), IKKa, cytoplasmic and nuclear NF-κBp65 were determined using Western blotting in K562/IR cells following a 48-h exposure to 0.5 µg/ml BBD9 or 8 µg/ml BBM. Flow cytometry was used to analyze the cell viability, apoptosis and necrosis; Western blotting was employed to determine the expressions of PARP, caspase-3, caspase-9 and LC3II in K562/IR cells exposed to different concentrations of BBD9 for 48 h. In nude mouse models bearing K562/IR cell xenograft, the tumor weight, tumor regression, and body weight changes of the mice were measured after treatments with 15 mg/kg and 30 mg/kg BBD9 and 100 mg/kg imatinib.

RESULTSThe IC50 of BBD9 and BBM was 0.73 µg/ml and 5.43 µg/ml, respectively. In K562/IR cell cultures, the expressions of p210(Bcr-Abl), IKKa and nuclear NF-κB p65 were all decreased following BBD9 and BBM treatments, but BBD9 produced more potent effect; cytoplasmic NF-κB p65 showed no obvious changes after the treatments. The cell apoptosis and necrosis increased with the concentrations of BBD9, which also dose-dependently increased the levels of cleaved caspase-3, csapase-9, PARP, and LC3II expression. In the tumor-bearing mouse model, BBD9 showed stronger effects than imatinib in reducing the tumor weight, promoting tumor regression, and increasing the body weight.

CONCLUSIONBBD9 can effectively inhibit the growth of K562/IR cells in vitro and in vivo by activating cell apoptosis, necrosis and autophage pathways, down-regulating expressions of p210(Bcr-Abl) and IKKa and suppressing the cytoplasm-to- nucleus translocation of NF-κBp65.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Benzamides ; Benzylisoquinolines ; pharmacology ; therapeutic use ; Drug Resistance, Neoplasm ; Female ; Fusion Proteins, bcr-abl ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; I-kappa B Kinase ; metabolism ; Imatinib Mesylate ; K562 Cells ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; Mice ; Mice, Nude ; Piperazines ; pharmacology ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; Pyrimidines ; pharmacology ; Transcription Factor RelA ; metabolism ; Xenograft Model Antitumor Assays