Objective To analyze the immunogenicity of selected B-cell epitopes of Epstein-Barr virus (EBV) latent membrane protein-2 (LMP2). Methods Three potential dominant B-cell epitopes of LMP2199-209, LMP2318-322 and LMP2381-391 from EBV LMP2 had been predicted using bioinformaties methods. The gene fragments of three epitopes were cloned respectively into pET32a(+) vector and transformed into E. coli strain BL21 (DE3). After identification by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting, the expression products were purified by Ni-NTA agarose affinity chromatography. BALB/c mice in immunized groups were immunized by multi-point intracutaneous injection with the three purified epitope proteins,respectively; and mice in control groups were injected with pET32a (+) protein or phosphate buffered saline(PBS), respectively. The sera from mice at week O, week 3 and week 6 of injection were collected for determination of epitope-specific antibody IgG by enzyme linked immunosorbent assay (ELISA) using epitope proteins as coating antigens. The ability of serum antibody recognizing nature EBV antigen was determined at week 6 of immunization. Results Three epitope proteins of LMP2199-209 ,LMP2318-322 and LMP2381-391 were successfully expressed in prokaryotic system. Epitopespecific antibodies IgG could be detected respectively in the sera of all immunized mice, and the levels of antibodies increased with immunized time increasing. The antibody levels in LMP2318-322 immunized group at week 3 and week 6 were significantly higher than that of pET32a (+) protein control group (F= 493.85 and 773.99, respectively; both P＜0. 05), and the antibody levels in LMP2381-391 immunized group at week 3 and week 6 were also significantly higher than that of pET32a (+) protein control group (F= 926.33 and 309.14, respectively; both P＜0.05). Antibody level in LMP2199-209 immunized group at week 6 was significantly higher than that of pET32a ( + ) protein control group (F=87.27, P＜0.05). The antibody IgG in serum from immunized mice with three epitope proteins could all recognize nature EBV antigens, especially LMP2199-209 and LMP2381-391 immunized groups.Conclusions Three possible dominant epitopes of LMP2199-209, LMP2318-322 and LMP2381-391 from EBV LMP2 are prepared by prokaryotic expression system and exhibit obvious immunogenicity, which could be used for further research of EBV infection and related tumor vaccine.

Items of design and signature from clinical trial drugs were made , the occurrence ratio of every item in 2265 informed consent form ( ICF) from 63 clinical trial drugs in Beijing Youan Hospital affiliated to Capital U-niversity of Medical Sciences were analyzed , and items with lower occurrence ratio were explored .Generally , the design and signature of ICF met the requirement of GCP .However , there were some defects of ICF and antonym of signature .Based on the analysis on the problems , some measures were put forward:formulate relevant standard op-erating procedures , strengthening the management of informed consent , and strengthen the system construction and education training , promote the hospital ethics construction , to protect the rights and interests of the subjects .

AIM To determine the contents of polymethoxylated flavones in Citri Reticulatae Pericarpium from eleven cultivar origins.METHODS HPLC was applied to screening extraction solvents (methanol,anhydrous ethanol,95％ ethanol,ethyl acetate and petroleum ether).Ultraviolet-visible spectrophotometry was adopted in the content determination of polymethoxylated flavones in eighteen batches of samples,nobiletin was taken as a reference substance,and the detection wavelength was set at 330 nm.RESULTS Ethyl acetate exhibited the best extraction effect,so it was selected as extraction solvent.Nobiletin showed a good linear relationship within the range of 3.024-13.104 μg/mL (R2 =0.999 8),whose average recovery was 101.37％ with the RSD of 1.34％.The contents of polymethoxylated flavones had obvious differences among samples from different cultivar origins,which were relatively high from Citrus reticulata ‘ Chachi’,Citrus reticulata ‘ Dahongpao’,Citrus reticulata ‘ Tangerina’,Citrus reticulata ‘ Ponkan’,Citrus reticulata ‘ Kinokuni’and Citrus reticulata ‘ Shiyueju’(more than 0.4％).CONCLUSION This simple and accurate method can effectively eliminate the interference of another flavonoids,which is suitable for the content determination of polymethoxylated flavones in Citri Reticulatae Pericarpium.

Objective:To investigate the relationship between previous bleeding history and poor prognosis of patients with acute upper gastrointestinal bleeding.Methods:This study was a prospective multicentre real-world study (Acute Upper Gastrointestinal Real-word study, AUGUR study). The data of patients with UGIB who were admitted to the emergency department of 20 tertiary hospitals in China from June 30, 2020 to February 10, 2021 were collected. According to the number of previous bleeding history, the patients were divided into three groups (0 time, 1-3 times, and≥4 times). Based on the patient’s demographic data, clinical characteristics, laboratory data, treatment, and outcomes, univariate and logistic regression analysis were performed to investigate the correlation between the number of previous bleeding and the 90-day mortality and rebleeding of patients with gastrointestinal bleeding.Results:A total of 1 072 patients with acute UGIB were included in this study. The all-cause mortality and rebleeding rate of all patients were 10.9% (117/1 072) and 11.8% (129/1 072), respectively. Among them, 712 patients (66.42%) had no previous bleeding, 297 patients (27.71%) had previous bleeding 1-3 times, and 63 patients (5.88%) had previous bleeding≥4 times. In univariate analysis, age, vital signs and consciousness on admission, history of liver cirrhosis, onset with hematemesis, admission hemoglobin, varicose veins bleeding, peptic ulcer bleeding, red blood cell infusion, tracheal intubation and the use of vasopressors after admission were risk factors for the 90-day mortality and rebleeding rate. Multivariate logistic regression analysis showed that patients with previous bleeding≥4 times had a higher risk of the 90-day mortality ( OR=2.17, 95% CI: 1.04-4.57, P=0.040) and rebleeding ( OR=2.32, 95% CI: 1.19-4.53, P=0.013). Conclusions:The history of previous bleeding≥ 4 times can be used as an independent risk factor for the 90-day mortality and rebleeding in patients with acute UGIB.

Objective To establish an appropriate diabetic retinopathy (DR) risk assessment model for patients with type 2 diabetes mellitus (T2DM).Methods A retrospective clinical analysis.From January 2016 to December 2017,753 T2DM patients in the Third Affiliated Hospital of Southern Medical University were analyzed retrospectively.Digital fundus photography was taken in all patients.Fasting plasma glucose (FPG),HbA1c,total bilirubin (TB),blood platelet,total cholesterol (TC),triglyceride (TG),high density lipoprotein cholesterol (HDL-c),low density lipoprotein cholesterol (LDL-c),apolipoprotein-A (apoA),apolipoprotein-B (apoB),serum creatinine,blood urea nitrogen (BUN),blood uric acid,fibrinogen (Fg),estimated glomerular filtration (eGFR) were collected.The patients were randomly assigned to model group and testify group,each had 702 patients and 51 patients respectively.Logistic regression was used to screen risk factors of DR and develop an assessment scale that can be used to predict DR.Goodness of fit was examined using the Hosmer-Lemeshow test and the area under the receiver operating characteristic (ROC) curve.Results Among 702 patients in the model group,483 patients were DR,219 patients were NDR.The scores for DR risk were duration of diabetes ≥4.5 years,4 points;total bilirubin ＜6.65 mol/L,2 points;apoA≥ 1.18 g/L,2 points;blood urea≥6.46 mmol/L,1 points;HbA1c ≥7.75％,2 points;HDL-c＜ 1.38 mmol/L,2 points;diabetic neplropathy,3 points;fibrinogen,1 point.The area under the receiver operating characteristic curve was 0.787.The logistic regression analysis showed that the risk factors independently associated with DR were duration of diabetes (β=1.272,OR=3.569,95％CI 2.283-5.578,P＜0.001),TB (β=0.744,OR=2.104,95％CI 1.404-3.152,P＜0.001,BUN (β=0.401,OR=1.494,95％CI 0.996-2.240,P=0.052),HbA1c (β=0.545,OR=1.724,95％CI 1.165-2.55,P=0.006),HDL-c (β=0.666,OR=1.986,95％CI 1.149-3.298,P=0.013),diabetic nephropathy (β=1.151,OR=3.162,95％CI2.080-4.806,P=0.013),Fg (β=0.333,OR=1.396,95％CI 0.945-2.061,P=0.094).The risk model was P=1/[1+exp-(-3.799+1.272X1+0.744X2+0.769X3+0.401X4+0.545X5+0.666X6+1.151X7+0.333X8)].X1=duration of diabetes,X2=TB,X3=apoA,X4=BUN,X5=HbA1c,X6=HDL-c,X7=diabetic nephropathy,X8=Fg.The area under the ROC curve was 0.787 and the Hosmer-Lemeshow test suggested excellent agreement (x2=10.125,df=8,P=0.256) in model group.The area under the ROC curve was 0.869 and the Hosmer-Lemeshow test suggested excellent agreement (x2=5.345,df=7,P=0.618) in model group.Conclusion The area under the ROC curve for DR was 0.787.The duration of diabetes,TB,BUN,HbAlc,HDL-c,diabetic nephropathy,apoA,Fg are the risk factors of DR in T2DM patients.

OBJECTIVETo screen a new strain which can transform panaxadiol saponins into the rare ginsenoside compound K.

METHODThe total saponins in stems and leaves of Panax notoginseng was used as a substrate in the liquid state fermentation process, and the results were detected by TLC and HPLC-ELSD to screen a strain from twelve plant pathogenic fungi which can produce ginsenoside compound K.

RESULTFusarium moniliforme was found to transform the total saponins to ginsenoside compound K efficiently in the all twelve fungal strains. In the fermentation process, ginsenoside Rb1 was transformed almost completely, and the content of ginsenoside Rd was decreasing evidently.

CONCLUSIONF. moniliforme is selected as a new high-yield strain. It is expected to be used to produce the high activity infrequent ginsenoside compound K and to improve the content of active principles in medicinal plants.

Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Fungi ; pathogenicity ; Ginsenosides ; chemistry ; Panax notoginseng ; chemistry ; microbiology

Objective:To comparatively analyze the diagnostic value of patent foramen ovale and the degree of right-to-left shunt among the traditional Valsalva maneuver (VM), the modified Valsalva maneuver (mVM) and the compression of the inferior vena cava combined with modified Valsalva maneuver (CM).Methods:A total of 132 patients with migraine, dizziness, chest pain and other symptoms in Longgang District People′s Hospital of Shenzhen from January 2021 to December 2021 were prospectively enrolled. The detection rate of patent foramen ovale and the degree of right-to-left shunt were assessed among the rest state and after performing the auxiliary maneuvers including VM, mVM and CM respectively.Results:After performing the auxiliary maneuvers, the detection rate of patent foramen ovale was significantly increased (88.6% vs 59.8%) with statistical significance (χ 2=36.026, P<0.001). There was a significant difference in the degree of right-to-left shunt among the three auxiliary maneuvers (χ 2=25.581, P<0.001). CM was the most effective for the detection of the large right-to-left shunt, and the difference was statistically significant compared with VM and mVM ( P<0.001, P=0.002). Conclusions:Auxiliary maneuvers is crucial for the diagnosis of patent foramen ovale. Compared with VM and mVM, performing CM can further improve the detection rate of patent foramen ovale and increase the degree of right-to-left shunt.

Objective:To observe the effect of fractional flow reserve derived from computed tomography(FFRcT)measurement system based on iterative algorithm in diagnosing survival prognosis of patients with acute ST-segment elevation myocardial infarction(STEMI)after they underwent emergency percutaneous coronary intervention(PCI).Methods:A total of 98 patients with acute STEMI complicated with multi vessel disease(MVD)were prospectively selected from the Affiliated Hospital of Inner Mongolia Medical University during October 2022 and February 2024.All patients were divided into two groups according to random double-blind method:FFRCT guidance group(49cases)and coronary angiography(CAG)guidance group(49cases).The FFRCT guidance group used FFRCT as the diagnostic basis,which carried out complete revascularization strategy according to FFRCT examination results.CAG guidance group handled infarct-related vessels as the CAG guidance.The basic clinical data,operation details of PCI surgery/technical indicators and utilization indicators of related resource of the two groups were observed,and angina pectoris attack and major adverse cardiovascular events(MACE)and other prognosis indicators of related survival of the two groups were observed and followed up.The primarily observed endpoints were all-cause death,heart failure,stroke,MACE,and composite endpoint event of stroke and revascularization again within 24 months follow-up.Results:The proportion of undergoing PCI on non-infarct-related vessel was 55.10％(27/49)in FFRCT group.In the comparison of operation detail of surgery/technique indicator after PCI surgery between two groups,the operation time of FFRCT guidance group was(65±10)min,which was longer than that[(60±8)min]of CAG guidance group,and the total length of stent was(31±10)mm,which was shorter than that[(36±10)mm]of CAG guidance group.There were statistically significant differences in the above two indicators between the two groups(t=-4.062,-6.710,P＜0.05),respectively.There were no statistically significant differences in the amount of contrast agent,average diameter of stent and length of stay between the two groups(P＞0.05).Compared with the CAG group,the incidences of non-lethal myocardial infarction,primary endpoint events,recurrent angina pectoris,and revascularization of FFRCT group significantly reduced within 24 months follow-up,and the differences of them between two groups were statistically significant(x2=4.926,5.273,6.091,5.804,P＜0.05),respectively.Conclusion:Using FFRCT measurement system based on iterative algorithm in PCI process of emergency for patients with acute STEMI complicated with MVD to examine and guide revascularization is safe and effective,which is worthy of recognition.It has obvious advantages over CAG guidance in reducing the risk of cardiovascular and cerebrovascular events,and in improving prognosis after PCI.

The passing of ethical review is a necessary conditions and prerequisite for the development of life science and medical research involving humans. At present, some medical and health institutions have no or insufficient ethical review capabilities. The lack of ethical review ability has become a bottleneck restricting the development of life science and medical research involving humans. According to documents such as Opinions on Deepening the Reform of the Review and Approval System and Encouraging the Innovation of Pharmaceutical and Medical Devices, Opinions on Strengthening the Ethical Governance of Science and Technology, institutions can entrust competent institutional ethics review committees or regional ethics review committees in writing to conduct ethical review. Entrustment ethical review provides a viable solution for institutions that need to carry out life science and medical research involving humans but do not have an ethics (review) committee or the ethics (review) committee is not competent to review. To conduct the entrustment ethical review, the entrustment between the principal and the trustee is required. According to The Measures for Ethical Review of Life Sciences and Medical Research Involving Humans, if medical and health institutions and their ethical review committees do not accept the formal entrustment to provide the ethical review opinions for other institutions, the local health authorities at or above the county level will impose administrative penalties and sanctions on the relevant institutions and personnel in accordance with the law. Signing the entrustment ethical review contract, implementing legal compliance entrusted ethical review to protect the rights and interests of the trustee and the principal, and protect the research participants.