Objective To optimize the sulfation-conditions of Bletilla striata polysaccharide (BSPS). Methods BSPS was sulfated with chlorsulfonic acid-pyridine. Reagent proportion, reaction temperature, and reaction time were selected with substitution degree and yield as index by L_9(3~4) orthogonal design. Results Results showed that the condition with 6∶1 volume ratio of pyridine and chlorine sulfonic acid, reaction temperature of 85 ℃, reaction time of 2 h was the optimum. Conclusion The optimal method for sulfation of BSPS is feasible, and it provides valuable parameters for further enlarged test.

The therapy to different type of breast cancer has resulted in chemoresistance which is induced by auto-phagy.Autophagy, to tumor’s generation, is a double edged sword, and it can result to chemoresistance and auto-phagic cell death through interaction with apoptosis in breast cancer chemotherapy.However, autophagy may be unrelated to chemoresistance.It will be a key research point to further explore the relationship and mechanism be-tween autophagy and chemoresistance.

AIM To study the expression, purification and activity assay of recombinant mouse protein kinase CK2? subunit from Escherichia coli. METHEDS The recombinant plasmid containing mouse protein kinase CK2? subunit cDNA constructed successfully was transformed into Escherichia coli BL21 (DE3) and specifically induced by IPTG. The recombinant mouse CK2? subunit was sequentially purified by DE 52, P11 phosphocellulose and Heparin Sepharose chromatography. The purified recombinant protein was analysed by SDS PAGE. RESULTS One protein with molecular mass of 42 ku was overexpressed by inducing ITPG. The recombinant protein was composed of approximately 30 6% of the total bacterial proteins. From 278 mg soluble proteins, the yield of the CK2? protein was 4 7 mg. SDS PAGE analysis of the purified recombinant protein showed only one band in agreement with native mouse CK2? subunit. The recombinant mouse CK2? and ? subunits were mixed at the same molar ratio. The produced CK2 holoenzyme displayed full activity. The characteristics and functions of reconstituted CK2 holoenzyme were consistent with those of the given native CK2. CONCLUSION The recombinant protein is mouse protein kinase CK2? subunit.

Objective To analyze the effectiveness of doctor-patient communication course on improvement of doctor-patient communication ability.Methods Totally 360 eight-year program medical students of clinical medicine from Shanghai Jiaotong University School of Medicine were chosen as the research object and were divided into educational group(n＝l80) and control group(n＝180).Students in educational group study doctor-patient communication course while those in control group do not.SEGUE Framework was used to conduct exploratory research between educational group and control group.Accuracy of questionnaire database was examined by Excel software and logical test.KoImogorov-Smimov(K-S test) was applied to analyze the scores and (x ± s) was used to express the data through SPSS 16.0 statistical software.Comparison between group differences are statistically significant(P＜0.05).Results Overall score was higher in educational group than in control group.Concerning of communication initiation,infomation collection and understanding of patients,scores of educational group were significantly higher than those of control group(P=0.001,P=0.002 and P=0.007,respec-tively).Furthermnore,KoInogorov-Smimov(K-S test) and (x ± s) statistical results showed that scores of detailed items were higher in educational group than in control group as well.Conclusions Doctor-patient communication course is of great importance to help medical students practice physician-patient communication based on related knowledge learning,but course content setting remains to be further improved.

Objective To study the expression of phosphorylated serine 910 of focal adhesion kinase(FAKps910)in circulation endothelial cells(CEC)from patients with hemorrhagic fever with renal syndrome(HFRS).Methods Fifty HFRS patients were involved in the study and divided into mild(n=17),moderate(n=20)and severe(n=13)groups according to patients' conditions. Twenty healthy volunteers were enrolled as control group. CEC were isolated by Percoll density gradient centrifugation method. The expression of FAKps910 in specific APC-CD31+ CEC was measured by flow cytometry. The data were analyzed by one-way ANOVA. Results The positive rates of FAKps910 in CEC were 59.87%±9.58% in early febrile stage patients and 21.14%±2.53% in the healthy controls, while it was 11.64%±2.17%in the later febrile or hypotension stage patients(F=262.31,P＜0.01).It gradually restored to normal afterwards. The positive rates of FAKps910 in CEC were 17.45%±2.64%,13.84%±2.54% and 7.47%±2.57%,respectively in mild, morderate and severe groups in the later febrile or hypotension stage, which were all significantly different from that in control group(F=52.642,P＜0.01).Conclusions FAK in CEC iS related with the clinical severity of HFRS patients. FAK may be involved in intergrin β3-mediated endothelial injury during Hantavirus infection.

Objective:To explore the effect of delphinidin on breast cancer and the underlying mechanisms.Methods:Human epidermal growth factor receptor-2 (HER-2) positive breast cancer cells MDA-MB-453 were treated by delphinidin.Proliferation of MDA-MB-453 cells was detected by CCK-8 after 48 h.TdT-mediated dUTP nick end labeling (TUNEL) assay and Western blot were used to explore apoptotic status for MDA-MB-453 cells.Fluorescence dot assay,immunofluorescence,and Western blot were used to identify autophagy in breast cancer cells.Results:Delphinidin suppressed proliferation of MDA-MB-453 cells.Delphinidin increased the number of TUNEL positive cells.Delphinidin downregulated the expression of caspase-3 and caspase-9,while upregulated the expression of cleaved caspase-3 and cleaved caspase-9 in a dose-dependent manner.Delphinidin enhanced the number of GFP-LC3 punctate dots,LC3 immunofluorescence dots and the expression of LC3-Ⅱ and ATG5.Delphinidin inhibited the expression of proteins in mTOR signaling pathway,induding AKT,mTOR,eIF4E and p70s6k.Conclusion:Delphinidin induced apoptosis and autophagy by inhibition of AKT/mTOR pathway in HER positive breast cancer cells.

Objective To investigate genetic characteristics of Neisseria gonorrhoeae (N.gonorrhoeae) isolates from Guangzhou city in 2014,and to analyze the relationship of N.gonorrhoeae multi-antigen sequence typing (NGMAST) sequence types (STs) with ciprofloxacin resistance.Methods An agar dilution method was used to determine the minimal inhibitory concentration (MIC) of ciprofloxacin in 97 N.gonorrhoeae isolates from Guangzhou city.PCR was performed to amplify the gyrA,parC,porB and tbpB genes from these isolates,followed by gene sequencing and determination of NG-MAST STs.Results Of the 97 N.gonorrhoeae isolates,95 (97.9％) were resistant to ciprofloxacin,including 41 high-level (MIC ≥ 16 mg/L) and 54 low-level (1 mg/L ≤ MIC ＜ 16 mg/L) resistant strains.Mutations were detected at codons 91 and 95 encoding serine in the gyrA gene of all the 95 ciprofloxacin-resistant strains,and in the parC gene of 93 resistant strains.The frequency of the mutation at codon 87 in the parC gene was 85.4％ (35/41) in high-level resistant strains,significantly higher than that in low-level resistant strains (59.3％[32/54],x2 =7.64,P ＜ 0.05).MAST STs were successfully determined for all the 97 N.gonorrhoeae isolates except 1 isolate with incorrect PCR amplicons.Of the 96 genotyped isolates,50 were assigned to 35 known STs by using the NG-MAST website (www.ngmast.net),among which,10 STs each contained 2 to 4 isolates.The most common ST was ST5309.Phylogenetic tree analysis revealed that the 96 genotyped N.gonorrhoeae isolates could be classified into 2 groups,and the proportion of isolates with MIC ≥ 16 mg/L is 46.4％ (39/84) in group 1,but only 1/12 in group 2 (x2 =6.27,P=0.012).Conclusions High-level resistance of N.gonorrhoeae to ciprofloxacin may be mainly associated with the mutation at codon 87 in the parC gene.NG-MAST STs may be related to the degree of ciprofloxacin resistance.

AIM:To examine the effects of thromboxane A 2 receptor ( TXA2 R) , the downstream product of cy-clooxygenase-2 (COX-2), on the proliferative ability and COX-2 expression in rheumatoid arthritis (RA) synovial cells. METHODS:The effects of TXA2 R antagonist SQ29548 and agonist U46619 on the proliferation of RA synovial cell line MH7A were detected by MTS cell proliferation assay , and their effects on COX-2 mRNA expression in MH7A cells were al-so examined by real-time PCR.In addition, the possible effect of U46619 on the proliferation of MH7A cells, when COX-2 was knocked down by siRNA , was determined by BrdU cell proliferation assay .RESULTS:SQ29548 inhibited the cell proliferation and the mRNA level of COX-2 while U46619 enhanced them.Moreover, U46619 reconstitute the proliferative ability of MH7A cells to some extent that inhibited by COX-2 siRNA.CONCLUSION: In RA synovial cells, TXA2R is able to control COX-2 expression, while it also mediates the effects of COX-2, suggesting that TXA2R might be an ideal candidate for RA treatment .

Objective:To analyze the clinical characteristics and diagnosis and treatment of children with Kasabach-Merritt phenomenon (KMP).Methods:A retrospective analysis was conducted on the clinical data and follow-up data of 8 patients diagnosed KMP in Beijing Children′s Hospital, Capital Medical University from January 2016 to January 2019.The clinical data included laboratory examination, diagnosis, treatment and prognosis.Results:Among the 8 children with KMP, 6 cases were male and 2 cases were female.The median onset age was 4 (0-17) months, 2 cases of neonatal onset.The median onset to the diagnosis time was 59 (34-140) days; 6 cases with bone destruction; 6 cases had misdiagnosis and mistreatment history, they were misdiagnosed as idiopathic thrombocytopenic purpura, Evans syndrome, abnormal bone and joint development; 4 cases were Kaposiform hemangioendothelioma; 8 cases were used alone or combined with the application of hormones, Sirolimus, and Vincristine, 7 patients underwent interventional therapy.All patients survived with a median follow-up period of 487 (112-1 033) days.Median time of platelet count returned to normal was 24.5 (7-60) days, and median time of fibrinogen returned to normal was 20 (7-30) days.Median time of D-dimer dropped to a normal was 105 (40-240) days.Conclusions:Children with concurrent platelet count and coagulation abnormalities should be considered with KMP.Doctors need to identify the potential visceral vascular lesions.Early diagnosis and treatment are important, which can improve the clinical prognosis of patients.

Objective To detect the effective connectivity of resting- state functional magnetic resonance imaging (fMRI) in normal adults. Methods 36 normal adults were performed resting-state fMRI scanning, and 5 brain netwokes were included as regions of interests. Independent component (ICA) was used to evaluate the effective connectivity, and multivariate Granger causality analysis (mGCA) was used to analyze the casuality between the networks. All preprocessing steps were carried out using Statistical Parametric Mapping 5.0 software. Results 5 classic resting brain networks including default mode network (DMN), memory network (MeN), motor network (MoN), auditory network (AN) and executive control network (ECN) were aquired. The mGCA presented significant casuality between DMN and other 4 networks, MeN and ECN, AN and MoN, ECN and AN. Conclusion There are specific brain effective connectivity of resting-state fMRI in normal adults, and there is significant causal link between these networks.