Objective:To develop a self-locked DNAzyme nanoprobe-based fluorescence amplification strategy for wash-free and ultrasensitive detection of breast cancer-derived small extracellular vesicles (sEV).Method:A DNAzyme self-locked probe was designed to recognize the epithelial cell adhesion molecule (EpCAM) specifically expressed on breast cancer-derived sEVs. Upon binding to EpCAM, the DNAzyme-lock structure was opened, restoring the DNAzyme cleavage activity. The activated DNAzyme then cyclically cleaved the RNA site on the substrate strand. Fluorescently labeled substrate strands were used to detect sEVs at varying concentrations, and the detection limit and linear range were determined.Results:The DNAzyme self-locked probe successfully identified breast cancer-derived sEVs and generated a fluorescent signal through cyclic cleavage. The proposed method achieved wash-free detection of sEVs, with the fluorescence intensity showing a strong linear correlation with sEV concentration ( R2=0.98). The linear detection range was 1.0×10 2-1.0×1.0 7 particles/μl, with a detection limit of 59 particles/μl. Conclusion:This study established a wash-free and highly sensitive strategy for quantifying breast cancer-derived sEVs, which provides a promising technical approach for the early diagnosis of cancer.

Objective:To develop a self-locked DNAzyme nanoprobe-based fluorescence amplification strategy for wash-free and ultrasensitive detection of breast cancer-derived small extracellular vesicles (sEV).Method:A DNAzyme self-locked probe was designed to recognize the epithelial cell adhesion molecule (EpCAM) specifically expressed on breast cancer-derived sEVs. Upon binding to EpCAM, the DNAzyme-lock structure was opened, restoring the DNAzyme cleavage activity. The activated DNAzyme then cyclically cleaved the RNA site on the substrate strand. Fluorescently labeled substrate strands were used to detect sEVs at varying concentrations, and the detection limit and linear range were determined.Results:The DNAzyme self-locked probe successfully identified breast cancer-derived sEVs and generated a fluorescent signal through cyclic cleavage. The proposed method achieved wash-free detection of sEVs, with the fluorescence intensity showing a strong linear correlation with sEV concentration ( R2=0.98). The linear detection range was 1.0×10 2-1.0×1.0 7 particles/μl, with a detection limit of 59 particles/μl. Conclusion:This study established a wash-free and highly sensitive strategy for quantifying breast cancer-derived sEVs, which provides a promising technical approach for the early diagnosis of cancer.

Objective To establish a HS-GC test method for the determination of enflurane,isoflurane,diflurane,sevoflurane and its metabolite hexafluoroisopropanol(HFIP)in blood,and to establish a LC-MS/MS method for the determination of trifluoroacetic acid(TFA),the co-metabolite of enflurane,isoflurane,and deflurane in blood.Methods Place 0.5 mL blood sample in a 10 mL headspace bottle,add 1.0 mL ultrapure water to mix,then add 0.5 mL n-butanol internal standard solution,sealed and heated at 70℃for 20 min,take the upper layer of gas for HS-GC analysis,qualitatived by dual-column retention time and quantified by the internal standard curve;Blood sample was acetonitrile precipitated protein,separated by liquid chromatography,scanned with electrospray ionization(ESI),negative ion mode,and examined in multiple reaction monitoring mode(MRM),qualitatived by retention time and characteristic ions,quantified by standard curve.Results The detection limits(LOD)of enflurane,isoflurane,desflurane,heptaflurane,HFIP and TFA are 0.05,0.2,0.5,0.05,0.5 μg/mL and 0.5 ng/mL,and linear range as 1～100 μg/mL(TFA:1～100 ng/mL),with correlation coefficients greater than 0.999.The extraction recovery rate is between 30%and 80%,and the intra-day and inter-day precision are less than 5%.The accuracy is between 85%and 115%.Conclusion This method is quick and simple,and can be used for qualitative and quantitative analysis of enflurane,isoflurane,desflurane,sevoflurane and their metabolites in blood.

Cholangiocarcinoma is a highly malignant disease with low surgical resection rate. It’s resistant to chemoradiotherapy and the prognosis of cholangiocarcinoma is poor. Although immune checkpoint inhibitors (ICIs) have achieved great success in tumor therapy in recent years, patients with cholangiocarcinoma have a poor response to immunotherapy, because of the complexity and diversity of immune microenvironment. Therefore, understanding the composition and characteristics of the immune microenvironment of cholangiocarcinoma, regulating targets in immune microenvironment, and adopting ICIs combined therapy, is important for immunotherapy for cholangiocarcinoma.

Gallbladder cancer is a high malignancy which is predisposed to invade adjacent organs and have lymph node metastasis. Gallbladder cancer is not sensitive to radiotherapy or chemotherapy with the worst prognosis among biliary tract cancers. At present, radical resection is the only possible method to cure gallbladder cancer. However, there are still many controversies about the surgical strategies, the extent of liver resection and lymph node dissection, and the treatment of incidental gallbladder cancer. In addition, under the background of the great success of immunotherapy and targeted therapy in a variety of solid tumors, it is also a question worthy of further considerations that whether the status of surgery in the treatment of advanced gallbladder cancer will be changed in the near future.

Objective:To construct a self-management behavior intervention scheme for diabetic patients based on Health Action Process Approach (HAPA) and to explore its application effects in patients with diabetes mellitus.Methods:By the convenient sampling method, patients with type 2 diabetes mellitus who were hospitalized in the First Hospital of Jilin University were selected as research objects. According to the random number table method, they were divided into the control group ( n=61) and the experimental group ( n=67) . Patients in the control group were given conventional nursing methods while patients in the experimental group were given the intervention scheme based on HAPA. Summary of Diabetes Self Care Activities (SDSCA) and Chinese version of Diabetes Distress Scale (DDS) were used to assess the self-management level and distress before and after intervention. Results:Before intervention, there were no statistically significant differences in SDSCA scores in all dimensions between the two groups ( P>0.05) . After intervention, SDSCA scores in all dimensions were higher in the experimental group than in the control group, and the differences were statistically significant ( P<0.05) . Before intervention, there were no statistically significant differences in DDS scores in all dimensions between the two groups ( P>0.05) . After intervention, DDS scores in all dimensions in the experimental group were lower than those in the control group, and the differences were statistically significant ( P<0.05) . Conclusions:The self-management intervention scheme based on HAPA can improve the self-management behavior of diabetic patients and reduce their psychological distress.

Objective To explore the predictive value of Tei index and plasma brain natriuretic peptide(BNP) on major adverse cardiovascular events(MACE) during long term follow up in the patients with anteriormyocardial infarction(MI).Methods A total of 238 patients with anterior MI were selected and divided into four groups according to the median of Tei index and BNP:G1 group(Tei≤0.66,BNP≤532.60 ng/mL,n=70),G2 group(Tei≤0.66,BNP＞532.60 ng/mL,n=51),G3 group(Tei＞0.66,BNP≤532.60 ng/mL,n=50) and G4 group(Tei＞0.66,BNP＞532.60 ng/mL,n=67).The follow up end was the MACE occurrence.Results There was a significant positive correlation between the Tei index and BNP(r=0.582,P=0.000).Tei entered the BNP regression equation(β=0.658,P＜0.05).The MACE-free survival rate had no statistical difference among 4 groups(x2 =9.975,P=0.019).The Cox regression analysis showed that Tei index and BNP were the independent predictive factors for MACE.Tei combined with BNP had stronger predictive value to MACE(AUC=0.781,95CI:0.721-0.841,P=0.000),its sensitivity was 72.52％ and specificity was 76.89％.Conclusion Tei combined with BNP has stronger predictive value for anterior MI during long term follow up.

Objective: To study the relationship between insulin resistance (IR) and coronary collateral circulation in patients with impaired glucose tolerance (IGT). Methods: A total of 227 patients with coronary angiography (CAG) were studied. There were 131 patients with male gender and the average patient’s age was (53.2 ± 11.0) years. IR (HOMA2-IR) index was measured by HOMA2 method, the severity level of coronary stenosis was assessed by Gensini scoring system, collateral circulation condition was determined by Rentrop classiifcation. 187 IGT patients were divided into 4 groups: Rentrop 0 group,n=55, Rentrop 1 group,n=42, Rentrop 2 group,n=39 and Rentrop 3 group,n=51; in addition, Control group,n=40 patients with normal glucose tolerance and coronary stenosis<50%. Results: Compared with Control group, all patients in 4 Rentrop groups had increased 2h-PBG, HbA1c, HOMA2-IR and Gensini score, while decreased fasting insulin (FINS), allP<0.05. Compared with Rentrop 3 group and Rentrop 2 group, the patients in Rentrop 1 group and Rentrop 0 group had elevated 2h-PBG, HbA1c, HOMA2-IR and Gensini score, while Rentrop 0 group had reduced FINS, allP<0.05. Multivariable regression analysis showed that HOMA2-IR index (R=0.518,P<0.05), HbA1c (R=1.916, P<0.05), 2h-PBG (R=2.130,P<0.05) and FINS (R=1.547,P<0.05) might be related to the severity of coronary stenosis. Binary regression analysis indicated that poor collateral circulation (the patients in Rentrop 0 group and Rentrop 1 group) was related to HOMA2-IR index (OR=1.679, 95% CI 1.101-2.558,P=0.016). Conclusion: HOMA2-IR index could be signiifcantly higher in patients with IGT combining chronic coronary occlusion. IR was the independent risk factor for the severity of coronary stenosis and coronary collateral formation.

OBJECTIVETo investigate the relationship between cytogenetic markers with World Health Organization (WHO) classification, disease progress and prognosis in cases with primary myelodysplastic syndromes (MDS).

METHODS298 patients with de novo MDS from the first affiliated hospital of medical school, Zhejiang University were enrolled in the retrospective analysis of WHO classification, karyotype, and prognosis. Follow-up study was also conducted.

RESULTSThe WHO classifications at first diagnosis were as follows: refractory cytopenia with unilineage dysplasia (RCUD), 18 cases; refractory anemia with ring sideroblasts (RARS), 8 cases; refractory cytopenia with multiline dysplasia (RCMD), 104 cases; refractory anemia with excess blasts-1, 76 cases; refractory anemia with excess blasts-2, 85 cases; MDS unclassified (MDS-U), 5 cases involved; and single del (5q), 2 cases. 39.6% of MDS patients carried karyotypic abnormalities. Among them, the frequency of numerical abnormalities, structural abnormalities and the existence of composite abnormalities were 45, 31, and 42, respectively. The composite abnormalities were unbalanced translocations and complex chromosomal abnormalities. The incidence of both karyotypic abnormalities and complex chromosomal abnormalities in RAEB group was higher than that in non-RAEB group (P<0. 05). An analysis based on IPSS-R Scoring System showed that advanced risk stratification (except the low-risk group) gradually enhanced the incidence of karyotypic abnormalities (P<0.05). In addition, the probability of evolution to leukemia increased with the higher IPSS-R score (P<0.05). In RAEB group, the cases with +8 chromosome, accounting for 19.5% of karyotypic abnormalities, had worse prognosis than those with normal chromosomes.

CONCLUSIONKaryotype was identified with an independent risk factor in MDS patients. Therefore, the information on cytogenetic analysis was critical for diagnosis, prognosis and individual treatment. MDS patients presenting+8 chromosome, an intermediate risk factor, were associated with a poorer outcome compared to cases with normal chromosomes in RAEB group.

Abnormal Karyotype ; Anemia, Refractory ; Chromosome Aberrations ; Chromosomes, Human, Pair 8 ; Follow-Up Studies ; Humans ; Karyotyping ; Myelodysplastic Syndromes ; Prognosis ; Retrospective Studies ; Risk Factors ; World Health Organization

Objective To understand the basic situation and existing problems of cost accounting information construction in the traditional Chinese medicine hospital. Methods We made a cross-sectional study of all traditional Chinese medicine hospitals cost accounting informatization, and then analyzed the results. Results In the survey of 1588 traditional Chinese medicine hospitals, there exists the problem with 78.65%for limited software;57.43%for not sharing between the systems;43.20%for outdated software. The occupancy rate was 81.86%for HIS system, 87.09%for the accounting software, 39.11%for cost accounting software, 5.42%for none. The proportion of hospitals was over 95% whose income data statistics to department;80% for spending data;73.05%for fixed assets depreciation cost;51.39%for amortization of intangible assets and 52.77%for medical risk fund. The proportion of hospitals was 81.49%whose data inside services statistics to medical auxiliary departments and higher than the logistics department, which was 63.85%.Conclusion The occupancy rate of cost accounting system software was limited in traditional Chinese medicine hospitals. The problems were limited cost accounting system software and unavailable information shared between the systems.