Objective:To investigate molecular mechanism of Helicobacter pylori(H.pylori)-mediated neutrophil apoptosis disorder.Methods:Neutrophils extracted from peripheral blood of healthy adults were used as research subjects,and H.pylori stan-dard strain NCTC11637 was added to culture system to construct a model of in vitro infection,which was divided into control group(with PBS or medium treatment),H.pylori group(H.pylori co-cultivated with neutrophils at ratio of 10∶1).After a certain time of in-fection,apoptosis-related molecules of neutrophils were detected by flow cytometry,ELISA and Western blot,respectively.Results:Cytokines IL-1β,IL-6 and TNF-α were secreted significantly higher in H.pylori group than control group,apoptosis rate was signifi-cantly lower than control group,apoptosis-related proteins Caspase-3 and Caspase-8 were higher than control group,but their activa-tion level were lower than control group.Conclusion:H.pylori infection may prolong neutrophil survival by inhibiting apoptotic pro-cess of neutrophils through inhibiting activation of Caspase-3/8.

Objective:To investigate molecular mechanism of Helicobacter pylori(H.pylori)-mediated neutrophil apoptosis disorder.Methods:Neutrophils extracted from peripheral blood of healthy adults were used as research subjects,and H.pylori stan-dard strain NCTC11637 was added to culture system to construct a model of in vitro infection,which was divided into control group(with PBS or medium treatment),H.pylori group(H.pylori co-cultivated with neutrophils at ratio of 10∶1).After a certain time of in-fection,apoptosis-related molecules of neutrophils were detected by flow cytometry,ELISA and Western blot,respectively.Results:Cytokines IL-1β,IL-6 and TNF-α were secreted significantly higher in H.pylori group than control group,apoptosis rate was signifi-cantly lower than control group,apoptosis-related proteins Caspase-3 and Caspase-8 were higher than control group,but their activa-tion level were lower than control group.Conclusion:H.pylori infection may prolong neutrophil survival by inhibiting apoptotic pro-cess of neutrophils through inhibiting activation of Caspase-3/8.

Objective:To investigate the role of miRNA-4298/PADI4/p53 signal axis in mediating 4f-induced apoptosis of leukemia cells.Methods:The cell growth density was observed under inverted microscope and the proliferation of leukemia cells was detected by CCK-8 counting assay. The expression of PADI4 and P53 at mRNA level was detected by qRT-PCR. Cell cycle and apoptosis were measured with flow cytometry. The expression of PADI4, P53, Bcl-2, Bax, caspase-3 and caspase-9 at protein level was detected by Western blot. Differential miRNA and mRNA expression profiles was detected by next generation sequencing. Databases such as TargetScan were used to predict the potential upstream and downstream genes of PADI4. A luciferase reporter assay was used to detect the 3′UTR of PADI4 targeted by miRNA-4298. Cell transfection assay was used to detect the effect siRNA, PADI4 vector, miRNA mimics and miRNA inhibitor in interference and rescue.Results:Nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor 4f could inhibit the proliferation of THP-1, K562 and U937 cells, and induce the apoptosis of these leukemia cells. It downregulated the expression of PADI4 mainly through the binding activity of miRNA-4298 to miRNA sponges, which resulted in the proliferation inhibition and apoptosis of leukemia cells. The inhibited proliferation and apoptosis of leukemia cells by 4f were associated with the increase of P53 expression after the decrease of PADI4 expression. The PADI4-dependent upregulation of P53 led to the ratio inversion of downstream Bcl-2/Bax, which activated caspase-3 or caspase-9 to induce the apoptosis of leukemia cells.Conclusions:The apoptosis of leukemia cells induced by Nrf2 inhibitor 4f was mainly associated with the miRNA-4298/PADI4/p53 axis, suggesting that it might be a novel signaling pathway for targeted therapy.

【Objective】 To analyze the related factors affecting perioperative blood transfusionin patients undergoing totalhysterectomy, explorethe effective measures to reduce allogeneic blood transfusion and provide data for the establishment of clinical blood transfusion single-disease evaluation index of total hysterectomy. 【Methods】 747 cases of total hysterectomy from three grade-A tertiary general hospitals and onematernal and child health care hospitalin Dalian wereselected, and divided into transfusion group(n=69)and non-transfusion(n=678). Detailed information was collected and analyzed, including patient demographics, operation information, blood routine before operation, postoperative recovery and transfusion volume. 【Results】 9.2%(69/747)of total hysterectomy patients received blood transfusion, with per capita red blood usageof(0.4±2.1)U. The age, operative time, volume of intra-operationbleeding, pre-operationhemoglobin(Hb) level, length ofhospital stay, types and days of antibiotics use between transfusion group and non-transfusion groupwere (49.7±9.1)vs(53±9.5)years old, (182.7±83.5)vs(119.5±64.8)min, (603±650)vs(160±173)mL, (96±26)vs(124±18)g/L, (9.3±4.8)vs(7.3±3.9)days, (2.2±1.1)vs(1.6±0.7)kinds, (6.0±3.9)vs (4.4±2.2)days, respectively, showing significant differences(P<0.05). Transfusion volumewas associated with volume of intra-operation bleeding (r=0.004), operative time (r=0.002) and Hb level of pre-operation(r=-0.022). 【Conclusion】 Blood management of patients undergoingtotal hysterectomy should be strengthened to reduce the incidence of allogeneic blood transfusion and improve patient outcomes by raising the pre-operation Hb level, shortening the operative time and reducing the volume of intra-operation bleeding.

Objective: To analyze the etiology and epidemiological characteristics of gastroenteritis virus in foodborne diseases from three cities in Shandong. Methods: From January to December 2017, six sentinel hospitals in Jinan, Yantai and Linyi city of Shandong Province were selected as the research sites. Stool samples of 1 397 diarrhea patients were collected, as well as basic information and clinical symptoms. Duplex quantitative RT-PCR was used to detect Norovirus genogroupⅠ (Nov GⅠ) and genogroupⅡ (Nov GⅡ), Sapovirus (SAV) and Human astrovirus (HAstV), respectively, quantitative RT-PCR was used to detect group A Rotavirus (RVA), and quantitative PCR was used to detect Enteric adenovirus (EAdV). The specific gene of the virus were sequenced and typed. It was compared that the gastroenteritis virus rate in cases with different characteristics and the clinical symptoms difference between the virus positive and negative cases. Results: The median age (P₂₅, P₇₅) was 23 (1, 42) , mainly male, 57.48% with 803 cased and children under 5 years old, 36.36% with 508 cases. The positive rate of gastroenteritis virus was 33.93% (474 cases), and that of Jinan, Linyi and Yantai City were 32.03% (147/459), 41.54% (189/455) and 28.57% (138/483), respectively (P<0.001). Nov GⅡ had the highest positive rate, 16.54% (231 cases), which, mainly GⅡ.P16/GⅡ.2 (48.28%, 56/116), peaked in May (24.75%, 50/202) and June (19.59%, 38/194). In patients of gastroenteritis virus positive, 44.51% (211/474) had vomiting symptoms, higher than that of patients of gastroenteritis virus negative (34.13%, 315/923). The difference was statistically significant (P<0.001). Conclusion In Shandong Province, the majority of gastroenteritis patients were male and children under 5 years old. Nov GⅡ possessed highest epidemic intensity, and peaked in spring and summer. Viral gastroenteritis had atypical clinical symptoms.

Objective To evaluate the survival and prognostic factors of esophageal cancer treated with definitive ( chemo ) radiotherapy by applying novel radiation techniques including three-dimensional conformal radiotherapy (3DCRT) or intensity-modulated radiotherapy (IMRT). Methods Clinical data of 2762 patients with non-operated esophageal squamous cell carcinoma who underwent definitive ( chemo ) radiotherapy from 2002 to 2016 in 10 hospitals were retrospectively analyzed.The prognostic factors were also identified and analyzed. Results The median follow-up time was 60. 8 months. The 1-, 2-, 3-and 5-year overall survival (OS) of all patients was 71. 4％,48. 9％,39. 3％,and 30. 9％,respectively.The 1-,2-,3-and 5-year progression-free survival (PFS) was 59.5％,41.5％,35.2％,and 30％,respectively.The median survival was 23 months.The median time to progression was 17. 2 months.Multivariate analysis demonstrated that age, primary tumor location, clinical stage, tumor target volume, EQD2 and treatment mode were the independent prognostic factors for OS.Primary tumor location,clinical stage,tumor target volume and EQD2 were the independent prognostic factors for PFS. Conclusions In this first large-scale multi-center retrospective analysis of definitive ( chemo) radiotherapy for esophageal squamous cell carcinoma in China, the 5-year OS of patients with esophageal squamous cell carcinoma is significantly improved by 3DCRT, IMRT combined with chemotherapy drugs. However, the findings remain to be validated by prospective clinical trials with high-level medical evidence.

Objective To establish a real-time quantitative polymerase chain reaction (qPCR) assay for B-cell lymphoblastic leukemia according to individualized and specific immunoglobulin gene rearrangements in leukemia cells, and to use it for the monitoring of minimal residual disease (MRD) of B-cell lymphocytic leukemia. Methods The immunoglobulin gene rearrangements of bone marrow samples from 15 cases of B-cell lymphoblastic leukemia were analyzed with a validated European BIOMED-2 system, and the individualized and specific qPCR-based quantification of leukemic immunoglobulin gene rearrangements was established. Results Unique and specific gene rearrangements of immunoglobulin light and heavy chains were identified in 14 cases and Ig-qPCR based on these gene rearrangements had a sensitivity of 10-5 and high specificity which met the international criteria in 10 patients. Leukemia MRD quantification with immunoglobulin gene rearrangement-based qPCR was similar as compared with other MRD detection methods. Conclusion Immunoglobulin gene rearrangement-based leukemia MRD quantification is feasible, sensitive, specific, precise and much valuable for clinical decision of treatments in B-cell lymphoblastic leukemia.

Objective This study aimed to investigate the associations between adipose tissue distribution and risks of macro-vascular complications in type 2 diabetic patients. Methods The study included T2DM patients either previously diagnosed by questionnaire or newly diagnosed with fasting plasma glucose(FPG)≥7. 0 mmol/L and/or 2 hour postprandial glucose ( 2h PG )≥11. 1 mmol/L from residents older than 40 years from Jiading Community, Shanghai, China(n=10 375). Each participant had gone through anthropometric measurements, blood tests, vascular function tests including carotid intima-media thickness ( CIMT ) , ankle-brachial index ( ABI ) , branchial-ankle pulse wave velocity( baPWV) as well as carotid plaques. Results WHR quartiles was significantly associated with high risks for ten-year cardiovascular disease risks [ ASCVD high risk: OR:1. 17 ( 1. 05-1. 31 );Framingham high risk:1. 13(1. 00-1. 29)]. However, WHR has no significant relations with risks of increased carotid intima-media thickness( CIMT) , abnormal ankle-brachial index( ABI) , increased brachial ankle pulse wave velocity( baPWV) or carotid plaques. Conclusions In type 2 diabetic patients, abdominal adipose tissue has an independent role in high risk for ten-year cardiovascular diseases. However it has no relations with other vascular functions including increased CIMT, abnormal ABI, increased baPWV, or carotid plaque.

Objective To simultaneously determinate the contents of four active ingredients (Ephedrine Hydrochloride, Pseudoephedrine Hydrochloride, Amygdalin and Glycyrrhizinate) ofSan Ao Decoction by HPLC; To optimize the extracting process ofSan Ao Decoction.Methods The orthogonal test was employed. Volume of water, time of extraction and times of extraction were set as investigation factors; the transport rates of Ephedrine Hydrochloride, Pseudoephedrine Hydrochloride, Amygdalin and Glycyrrhizinate and the extraction rates were set as investigation indexes to conduct multi-indexes grading.Results The influence factors of the extraction ofSao Ao Decoction were the times of extraction > decoction time > the volume of water. The best extraction condition was: ten fold water; extract for three times; 1 h for each time.Conclusion The optimized extraction process is stable and feasible.

Objective To standardize the detection procedure and improve the detection ability through the capability evaluation of transfusion compatibility tests for blood transfusion departments in Dalian .Methods Both on‐site inspection and external quality assessment(EQA) were used .The items of on‐site inspection consisted of equipments ,reagents ,tests ,operating instructions and re‐cords .EQA included ABO grouping ,Rh(D) grouping ,antibody screening and crossmatching .Results 42 of 62 blood transfusion departments were qualified .Only one was unqualified in on‐site inspection because antibody screening were not carried out .The un‐qualified ratio of second‐class hospitals′ EQA was the highest (42 .3% ) .The coincidence rates of antibody screening and crossmatching were 82 .0% and 77 .4% respectively ,while those of ABO grouping and Rh(D) grouping was 100 .0% .Conclusion Relatively fixed staffing in laboratories and continual training was important for the improvement of transfusion compatibility tests .