Objective To study the value of magnetic resonance vessel wall imaging in assessing the atherosclerotic plaque burden of rabbit model.Methods We built up abdominal atherosclerotic animal model in 30 New Zealand rabbits by high lipid diet combined with abdominal artery denudation.The animals were divided into 3 groups randomly,which were the 1-week group,1-month group and 2-months group.The MRI and histology examination were carried out at relative time points.The correlations of area or thickness of vessel wall by MRI with histology examination were analyzed.Results Among the 30 rabbits,3 died due to anesthesia or surgery,one rabbit model failed because of the thin vessel,and another 3 died of diarrhea or inflammation during the high lipid diet feeding.Eventually,totally 23 rabbits fulfilled the examinations (7 in 1-week group,7 in 1-month group and 9 in 2-months group).The vessel wall area of histology examination grew larger along with the post-surgery duration,from 1.7663 mm2of 1-week group to 2.4371 mm2 of the 1-month group till 3.5978 mm2 of 2-months group,with statistic significant difference among 3 groups (F=5.052,P=0.017).There were strong correlations of area or thickness vessel wall resulted from MRI with histology examination(r=0.688,0.642;P=0.001,0.002).Conclusions High resolution MR vessel wall imaging technique may evaluate and follow up the plaque burden in the early stage of atherosclerosis.

Objective To explore the predictive value of serum CEA and cytokeratin-19 fragments (CYFRA21-1)prior treatment for the epidermal growth factor receptor (EGFR) mutation and efficacy of tyrosine kinase inhibitors ( TKI ) in patients with non-small cell lung cancer.Methods The study was a clinical research.Totally 101 matched tissue and plasma samples were collected from Peking University People′s Hospital from 2012 to 2013.All clinical specimens were analyzed for EGFR mutations in exons of 18, 19, 20 and 21 by ADx-ARMS and direct sequencing, and the serum levels of CEA and CYFRA21-1 were analyzed by ECLI.The correlation between EGFR mutant status and efficacy of EGFR-TKI and clinicopathological parameters were analyzed by χ2 test, Log-rank text and Cox proportional hazards regression model.Results The mutation rate was 60.4%(61/101) by ADx-ARMS and 33.7%(34/101) by direct sequencing.Mutations were more frequently observed in the higher serum CEA level patients(≥5μg/L,78.8%).However, the rates of EGFR mutations of different CEA levels were similar.Among the patients receiving TKI therapy, the efficacy of EGFR-TKI was closely related to serum CYFRA21-1 level prior treatment and EGFR mutation (χ2 =8.903, P =0.003; χ2 =28.590, P <0.001 ).And serum CYFRA21-1 level prior treatment and EGFR mutation were independent factors for EGFR-TKI treatment affecting PFS (RR=0.298, P<0.001;RR=0.086, P<0.001).Conclusion The mutation rate of EGFR was significantly related with the expression level of CEA prior treatment, and serum CEA and CYFRA21-1 levels prior treatment could be potential predictors of EGFR-TKI efficacy.

Objective:To observe the effects of low intensity pulsed ultrasound(LIPUS) on the osteogenic differentiation of rat bone marrow mesenchymal stem cells(BMSCs) on titanium surface.Methods:BMSCs from Wistar rat bone marrow were respectively cultured on the flat titanium surface and the large grain blast acid etched(SLA) titanium surface,and induced by mineralization medium.Then,the cells were interfered by LIPUS and a control condition.Alkaline phosphatase(ALP) were quantitative determinated after 3 and 7 d mineralization induction respectively,ALP staining were observed after 14 d induction.Alizarin red staining were observed after 21 d mineralization induction.Osteogenic related protein and gene expressions were detected after mineralization induction.Results:ALP in culture medium of LIPUS group was higher than that of the control group after 3 d and 7 d mineralization induction(P＜0.05).LIPUS group showed stronger ALP staining and alizarin staining,and more mineralized nodules than control group.The expression of osteogenic related proteins,including Runx2,BMP2,OPN in LIPUS group increased.Osteogenic related genes expression,including ALP,Runx2,BMP2,OPN,OCN and Col-1 of the LIPUS group increased.Conclusion:The osteogenic differentiation of BMSCs on the fiat titanium surface or SLA titanium surface can be promoted by LIPUS.

Objective To explore the correlations of diffusion-weighted imaging (DWI) types and the degree of neurologic impairment in acute ischemic stroke patients with atrial fibrillation.Methods DWI images and National Institutes of Health Stroke Scale(NIHSS) of 186 patients with acute ischemic stroke patients with atrial fibrillation were collected retrospectively.The correlation of DWI features and NIHSS was analyzed.Results On DWI,all acute ischemic stroke patients with atrial fibrillation presented high signal intensity.Single cortex-subcortical infarction mostly appeared in the anterior circulation(94,50.5%);Multi-infarction commonly occurred in the posterior circulation(18,13.0%);The neurological deficit scores of subcortical-cortex infarction in the left anterior circulation(16.75±7.10) were higher than that in the right side(13.50±5.70)(P<0.05).The neurological deficit scores of cortex-subcortical infarction in the posterior circulation (6.38±2.03) were significantly lower than that in the multi-infarction (16.77±8.90) (P<0.05).Conclusion DWI types are valuable for etiological diagnosis in ischemic stroke.Combination with NIHSS score could provide a basis for clinical individual treatment programs selection and prognostic evaluation.

Objective To investigate the relationship between waist circumference and new-onset non-alcoholic fatty liver disease in non-obese patients with diabetes mellitus. Methods A total of 1 950 patients with diabetes mellitus, who determined fasting plasma glucose(FPG)≥7.0 mmol/L or who were using hypoglycemic drugs and FPG<7.0 mmol/L,and body mass index (BMI)< 25 kg/m2, was selected in this study using prospective cohort method. Patients were divided into five groups according to the baseline data of waist circumference, including waist circumference<78 cm (A group, n=387), 78 cm90 cm (E group, n=421). Multiple Logistic regression model was used to analyze influential factors of new-onset non-alcoholic fatty liver disease in non-obese patients with diabetes mellitus. Re?sults The average duration of follow-up was(47.24±5.13) months. The incidence rate was 11.85%(231/1 950) in patients with non-alcoholic fatty liver disease. The incidence rates were 6.98%, 9.28%, 12.38%, 14.19%and 15.68%in A, B, C, D and E groups, and which were increased with the increased waist circumference (P<0.05). Results of multiple Logistic re?gression model analysis showed that compared with A group,OR values were 1.97 and 2.19 in D and E groups respectively (P<0.05). Conclusion Waist circumference≥85 cm was the risk factors of new-onset non-alcoholic fatty liver disease in non-obese patients with diabetes mellitus.

Objective To investigate the clinical characteristics of the human Adenovirus (HAdv) infections in allogeneic hematopoietic stem cell transplantation ( allo-HSCT) patients and explore the clinical significance of HAdv monitoring .Methods A total of 845 cases underwent allo-HSCT were included retrospectively in Perking University People′s Hospital from October 2012 to August 2014.Peripheral blood HAdv load were monitored twice weekly within 100 days after allo-HSCT, or whenever necessary quantitatively by real-time PCR. Meanwhile, other clinical samples such as stool , urine, and bronchoalveolar lavage fluid ( BLAF ) were also detected qualitatively whenever necessary .The follow-up period was at least six months after allo-HSCT.All clinical data were collected and analyzed .Results The total positive rate of HAdv was 3.4% ( 29/845 ) .The incidence of HAdv infection was higher in children [3.8%(6/155), <18y] than that of adults [3.3%(23/690),≥18y].HAdv infection diagnosed within 100 days after allo-HSCT accounted for 72.4%(21/29) of the total number of positive cases .There were 19 cases detected positive in peripheral blood , 16 cases in stool , 9 cases in urine , and 1 cases in BLAF , respectively.One patient was positive in peripheral blood , stool and urine.The overall median time of HAdv was 69 (13-189) d.The median time was 56 (53 -144) d in stool ,which was earlier than that of in peripheral blood , urine and stool.Among 29 cases of HAdv positive patients , 17 patients were coinfected with Cytomegalovirus(CMV) and 11 casess with Epstein-Barr virus(EBV).Twenty-five cases of HAdv were diagnosed with acute graft-versus-host disease(aGVHD) before HAdv infection, and 4 cases were diagnosed with chronic graft-versus-host disease ( cGVHD ) . The most common clinical manifestation was HAdv enteritis (14 cases), followed by hemorrhagic cystitis (7 cases).Two cases complicated with multiple organ injury ( >2 ) clinically, 1 cases with pneumonia.There were 8 cases of death at the end of follow-up.Conclusions HAdv is an important pathogen causing infection in patients after allo-HSCT. The infenction is characterized with multiple organ involvement .CMV and EBV coinfection is common .HAdv monitoring was of great significance in allo-HSCT patients.

OBJECTIVETo obtain 1-4 IgG-like domains of mouse vascular endothelial growth factor receptor 2 (VEGFR2) fusion protein (mVEGFR2D1-4/GST) and identify its antiginicity and biological activity.

METHODSThe gene of mVEGFR2D1-4 was amplified by RT-PCR from 14-days embryos of Balb/c mice. The PCR product was cloned into pET-42a prokaryotic expression vector to construct the recombinant plasmid pET-42a-mVEGFR2D1-4, which was transformed into E. coli BL21 (DE3) strain for mVEGFR2D1-4/GST expression. The fusion protein was identified by SDS-PAGE and Western blotting, and the antigenicity of the protein purified by affinity chromatography was characterized by ELISA. The VEGF blocking effect of the purified protein in human umbilical vein endothelial cells (HUVECs) were evaluated in in vitro cell cultures.

RESULTSThe mVEGFR2D1-4 gene was obtained, which had an identical sequence to that retrieved in GenBank. The prokaryotic expression vector for mVEGFR2D1-4 was successfully constructed as confirmed by enzyme digestion and DNA sequencing. Both Western blotting and ELISA demonstrated the antigenicity of the purified mVEGFR2D1-4 fusion protein, which obviously blocked the effect of VEGF in promoting HUVEC proliferation in vitro.

CONCLUSIONThe mVEGFR2D1-4/GST fusion protein obtained shows a strong antigenicity and biological activity to facilitate further study of active anti-tumor immunotherapy targeting VEGFR2.

Animals ; Cell Proliferation ; Escherichia coli ; genetics ; metabolism ; Female ; Gene Expression ; Genetic Vectors ; Human Umbilical Vein Endothelial Cells ; Humans ; Mice ; Mice, Inbred BALB C ; Plasmids ; Recombinant Fusion Proteins ; genetics ; immunology ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; immunology ; isolation & purification

This study was aimed to explore the curative effect of Zhuo-Du-Qing (ZDQ) particles on diabetic cystopa-thy (DCP) based on the theory of turbid toxin. A total of 100 DCP patients were randomly divided into the control group with 47 cases and the treatment group with 53 cases. Mecobalamin tablets was given in the control group. And ZDQ particles were given to the treatment group. The treatment course was 3 months. The observation was made on the bladder residual urine (BRU), clinical manifestation integral, hemodynamic indexes, HOMA-IR and HbA1c before and after the treatment. The results showed that compared with pretreatment, the BRU, clinical manifestation integral, hemodynamic indexes, and HOMA-IR were significantly reduced (P < 0.01). Compared with the control group, the BRU, clinical manifestation integral, hemodynamic indexes, and HOMA-IR were also significantly reduced (P <0.01). The effective rate in the treatment group was 81.1%, which was significantly higher than 59.6% in the control group (P< 0.05). There were no obvious changes on the blood routine examination, urine routine examination, stool routine examination, liver function, renal function, electrocardiogram and so on before and after treatment. It was con-cluded that the treatment of DCP with ZDQ particles was safe and effective. To remove toxin and descend turbid may be another effective treatment method for DCP. The occurrence and development of DCP were closely related to the turbid toxin.

Objective To investigate germline mutation of breast cancer susceptibility genes BRCA1/2,TP53 and PTEN in Chinese breast cancer patients. Methods All of128 female breast cancer patients in Peking University People′s Hospital from January 2016 to August 2018 were selected as objects. Among them,44 cases were sporadic breast cancer and 84 werebreast cancer patients with genetic high risks. Germline mutations of BRCA1,BRCA2,TP53 and PTENwere detected by NGS.χ2 test was used to analyze the difference of pathogenic mutation rates between sporadic breast cancer group and breast cancer with high genetic risks.Groups were divided according to the clinical features of the patients(family history, triple-negative breast cancer,age and bilateral breast cancer).Among them,there were 42 cases with family history of breast cancer,34 cases of triple-negative breast cancer,33 cases of early-onset breast cancer and 7 cases of bilateral breast cancer. Fisher′s exact probability test compared the relationship between pathogenic mutations of BRCA1/2 gene and clinical characteristics of breast cancer patients with hereditary risk factors. Results In 128 cases of breast cancer,30 germline mutations of BRCA1/2 were detected, including 13 pathogenic mutations and 3 newly discovered mutations(BRCA1:c. 4760C>G,BRCA2:c. 44134414del and BRCA2:c. 64826485del). The new mutations may be unique mutations of Chinese population. There were 3 cases of TP53 mutations,including 1 pathogenic mutation. All of the 3 mutations were found in early-onset breast cancer. Germline mutation of T53 has important detection significance for early-onset hereditary breast cancer. There were 5 cases of PTEN mutations,including 3 pathogenic mutations. Among 84 breast cancer patients with genetic high risks,the carry mutation rate was 40.5％(34/84)and the pathogenic mutation rate was 15.4(13/84). Among 44 sporadic cases,the carry mutation rate was 9％(4/44). The pathogenic mutation rate was 6.8％(3/44). Breast cancer susceptibility genes were carried at a higher rate in breast cancer patients with genetic high risks(P<0.001). BRCA1/2 mutations did not show statistical differences among groups of breast cancer patients with hereditary high risk factors . Conclusion Germline mutation detection of breast cancer susceptibility genes by next-generation sequencing is of great significance in breast cancer risk prediction and prognosis evaluation.

Objective: To analyze CYP2C9 and VKORC1 gene polymorphisms in Chinese Han population and their correlation with the maintenance dosage of warfarin. Methods: From October 2017 to April 2018, 458 Chinese Han patients (213 males and 245 females, aged from 26 to 94 years old) who underwent coagulation analysis in Peking University People′s Hospital were included in this retrospective study. PCR-Fluorescent probe method was applied to detect CYP2C9*3 and VKORC1-1639A>G gene polymorphisms in 458 patients, and among them, 130 patients who took warfarin for anticoagulant therapy and reached the international standard ratio of prothrombin time (INR) within the range of 2.0-3.0 were recorded. The basic information, dosage of warfarin and INR were also recorded. The statistical analysis data were compared with the reference table of recommended dosage of warfarin for different genotypes of patients recommended by FDA and the formula of predicted dosage of warfarin was simply verified by SPSS. Results: Among the 458 patients who took anticoagulant therapy, the genotype frequencies of CYP2C9*1/*1(AA), CYP2C9*1/*3(AC) and CYP2C9*3/*3(CC) were 90.8%, 8.5%, and 0.7%; the genotype frequencies of VKORC1-1639GG and VKORC1-1639AG were 0.9% and 14.2%; the genotype frequencies of VKORC1-1639AA was 84.9%. After INR was reached, the results showed that the variant CYP2C9*1/*3 and CYP2C9*3/*3 required lower daily maintain dosage [(2.92±1.29) mg] than wild-type CYP2C9*1/*1 patients did [(3.91±1.63) mg], with statistically significant difference (P=0.018). And variant VKORC1-AA required lower daily maintain dosage [(3.68±1.64) mg] than variant VKORC1-AG patients did [(4.54±1.29) mg], with statistically significant difference (P=0.001). The application dosage of warfarin in patients with different VKORC1+CYP2C9 genotypes was consistent with the recommended dosage of the FDA reference table. The prediction accuracy of miao 2007 formula was lower than that of IWPC formula, and 94.1% of patients′ dosages of warfarin were underestimated. Conclusion Patients with CYP2C9*3 or VKORC1-AA genotype required lower warfarin dosage. The CYP2C9 and VKORC1 gene polymorphisms had a certain correlation with maintenance dosage of warfarin.