Objective: To investigate the function of anti-PD-1 (scFv)/IL-15/IL-15Rα-sushi fusion protein (PD-S15) to specifically bind to PD-1 in vitro and to explore its effect on NK/T cell proliferation. Methods: The human anti-PD-1 (scFv) gene sequence and human IL-15/IL-15Rα-sushi fusion gene sequence were synthesized chemically. The recombinant expression plasmid pUC57-PD-S15 was constructed by enzyme digestion and ligation of the two target genes, and then transiently transfected into HEK293T cells by lipofectamineTM 2000. The supernatants of cell culture medium were acquired, and the expression of PD-S15 fusion protein in cell culture supernatants was detected by Wb assay. PBMCs and TILs were cultured in mediums with different proportion of PD-S15/X-VIVOTM15, respectively. Then, the capacity of PD-S15 fusion protein to bind to PD-1 in vitro and its effect on the proliferation of PBMCs and the proportion of CD3+CD8+, CD3+CD4+ and CD3-CD56+ subsets were detected by flow cytometry. The effect of PD-S15 fusion protein on the proliferation of TILs was detected by cytometry. Results: The successful construction of pUC57-PD-S15 eukaryotic expression plasmid was confirmed by double enzyme digestion and sequencing, and then successfully transfected into HEK293T cells. The relative molecular weight of the target protein was approximately 55 000, and was in line with expectations. PD-S15 fusion protein could specifically combine with PD-1 in vitro (P<0.05) and stimulate NK/T cell proliferation (P<0.05). Compared with classical TILs culture method, the efficiency of activation and amplification of T cells in vitro by PD-S15 culturemethodwasbetter (P<0.01). Conclusion: PD-S15 fusion protein can specifically target PD-1 and rapidly expand NK/T cells in vitro, which lays a foundation for the selective expansion of CD8+PD-1+ antigen-specific T lymphocytes from tumor tissues and even peripheral blood.

Objective To study the effectiveness of comprehensive control measures based on systematic ecological system construction to interrupt the transmission of schistosomiasis in hilly endemic regions in Sichuan Province ,so as to provide the ev?idence for adjustment of schistosomiasis prevention and control strategies. Methods A high endemic area of schistosomiasis, Panao Township of Dongpo District in Meishan City,was selected as a demonstration area. The comprehensive measures for schistosomiasis control with focus on systematic ecological management were implemented,and the income of residents,index?es of schistosomiasis control effect and so on were investigated before and after the intervention and the results were compared. Results The project based on systematic ecological system construction started in 2009 and 317.351 million Yuan was put into the construction. The construction included economic forest plant base(1 866.68 hm2,72.66%of the total farmland areas),eco?logical protection gardens(585.35 hm2)and so on. Totally 97.04% of historical areas with Oncomelania hupensis snails were comprehensively improved. In 2015,the peasants' pure income per capita increased 4 938 Yuan,with the average annual growth rate of 14.69%. All the farm cattle were replaced by the machine. The benefit rate of water improvement was increased by 52.84%and the coverage rate of harmless toilets increased by 18.30%. The positive rate of serological tests for schistosomiasis decreased from 7.69%to 3.50%,and the positive rate of parasitological tests decreased from 1.18%to 0. The area with snails was decreased from 23.33 hm2 to 0. The awareness rate of schistosomiasis control knowledge and correct behavior rate of the resi? dents increased from 85.50%and 82.60%to 95.70%and 93.90%respectively. Conclusions The comprehensive schistosomia?sis control measures based on systematic ecological management are conform to the currently actual schistosomiasis prevention and control work in hilly endemic regions,and have good ecological economic benefit and schistosomiasis control effectiveness, which provide an effectively new model of prevention and control for advancing process,consolidating the effect,finally realiz?ing goal of interruption and elimination of schistosomiasis in hilly endemic regions.

BACKGROUND:Increasing autologous stem cellmobilization is conceived to achieve effectively repair of cardiac ischemic injury. Therefore, it is important to seek a specific and effective mobilization agent. OBJECTIVE:To observe the effects of hypoxia-inducible factor-1α(HIF-1α) on bone marrow mesenchymal stem cellmobilization in myocardial infarction. METHODS:Left anterior descending artery was ligated to establish a rat model of acute myocardial infarction in 90 outbreeding Sprague-Dawley rats, and then the models were randomly divided into three groups. In HIF-1α-antisense oligonucleotide (ASODN) group, HIF-1α-ASODN was infused into the tail vein to restrain the expression of HIF-1αin infarcted ischemic tissue. In HIF-1α-missense oligonucleotide (MSODN) group or control group, an equal volume of HIF-1α-MSODN or saline was injected. RESULTS AND CONCLUSION:After 30 hours and 7 days of modeling, the number of bone marrow mesenchymal stem cells and expression of vascular endothelial growth factor in the peripheral blood of the control group were similar to the HIF-1α-MSODN group, but significantly higher than the HIF-1α-ASODN group. After 7 days of modeling, the expressions of HIF-1αprotein, vascular endothelial growth factor protein and mRNA in the ischemic myocardial tissues of the control group were similar to the HIF-1α-MSODN group, but significantly higher than the HIF-1α-ASODN group. After 7, 14 and 28 days of modeling, the capil ary density in the ischemic myocardial tissues of the control group was similar to the HIF-1α-MSODN group, but significantly higher than the HIF-1α-ASODN group. These findings indicate that after acute myocardial infarction, high expression of HIF-1αexhibits a causal relationship with mobilization of bone marrow mesenchymal stem cells, initiating a series of self-healing process of myocardial tissues.

Objective To investigate the incidence of contrast induced nephropathy (CIN) among different patient groups after contrast agent injection.Methods A total of 1243 patients were included in this study (male =694,female =549).The SCr level one week before and 72 hours after the CT examination and the incidence of CIN were recorded and comparison was made among groups according to sex,age,body mass index (BMI),the history of high blood pressure (HBP),diabetes mellitus (DM),chronic kidney disease (CKD),chronic heart failure (CHF),tumor,nephrotoxicity drug (NTD) usage.The frequency,type,dose and injection velocity of the contrast media(CM)were also recorded.Multivariate predictors of CIN were identified by Logistic regression using step-wise selection with entry and exit criteria of P ＜0.10,results were tabulated as odds ratios (OR) with 95％ confidence intervals (CI).Results Among 1243 consecutive patients,the incidence of CIN was 5.5％ (68/1243).Patients with a history of HBP,DM,CHF,CKD or tumor presented with higher incidence of CIN than that of controls (5.9％,51/868 vs.4.5％,17/375).CIN developed in 9 of 203 patients (4.4％,9/203) with CKD and in 59 of 1040 patients (5.7％,59/1040)without CKD.There was no significant difference between the two groups(x2 =0.51,P =0.30).In CKD (-) group,the incidence of CIN was higher in females,patients with DM and patients using LOCM than those of males,DM (-) and using low osmolality contrast medium (IOCM) (P ＜ 0.05),but there was no statistical significance in CKD (+) group.Logistic regression analysis showed that women,age ≥ 75 years,DM,LOCM,NTD,tumor,the time of using CM more than once per month were the most significant predictors of CIN (OR ＞ 1).Conclusion Women,age ≥ 75 years,LOCM,NTD,tumor,and the frequency of using CM more than once per month were more likely to develop CIN.

Objective To evaluate the feasibility of assessing osteoarthritis (OA) in hip dysplasia using 3D delayed gadolinium-enhanced MRI of cartilage (dGEMRIC).Methods Thirty-five hips in 20 patients with radiographic evidence of hip dysplasia underwent 3D-dGEMRIC scanning.Clinical symptoms were assessed with the Western Ontario and McMaster Universities Osteoarthritis ( WOMAC ) questionnaire.Radiographic measurement of lateral center-edge angle and T(o)nnis grading were performed on the X-rays.Hips of T(o)nnis grade 1were included in the group of hips with early OA,while the hips with no evidence of OA and without pain symptom were included in the group of hips with normal morphology.The 3D-dGEMRC scans were completed on a 1.5 T MR scanner.The data of 3D-dGEMRIC was reconstructed radically.The dGEMRIC indices were measured on six sites of periphery zones of hip cartilage on reconstructed images.The dGEMRIC indices among different groups were analyzed by non-parametric tests.The differences of dGEMRIC indices among six sites in the group of early OA or the group of normal morphology were analyzed by Wilcoxon test.Results The mean dGEMRIC indices of six sites were lower in group of T(o)nnis grade 1than in group of T(o)nnis grade 0 ( Z =- 2.149,P =0.032 ),and lower in group of T(o)nnis grade 2 than in group of T(o)nnis grade 1( Z =- 1.990,P =0.047 ).The dGEMRIC indices of the anterior site,anterosuperior site,superior-anterior site,and superior site were significantly different between the group of hips with early OA and the group of hips with normal morphology (Z =-2.333--2.041,all of the P values were lower than 0.05).In the group of hips with normal morphology,the dGEMRIC indices of superior-anterior site of hip were lower than superior site(P =0.028).In the group of hips with early OA,the dGEMRIC indices of superior-anterior site were lower than the other sites except for anterior-superior site ( Z =- 3.041- - 2.277,all of the P values were lower than 0.05 ).Conclusions 3 D-dGEMRIC might be a sensitive technique for detection of glycosaminoglycans alteration in early OA and staging of OA in hip dysplasia.Radial reconstruction could provide an accurate assessment of OA,and the results demonstrated that early cartilage alteration could be detected in the anterior to superior sites of hips,and the earliest cartilage alteration may occur in the superior-anterior site of hips.

Objective To detect the differences of grey matter volume between the patients with mental retardation (MR) presented clinically as operation deficit (OD) or as language deficit (LD) and the children with typical normal development using optimal VBM.The developmental connections between brain gray matter and language or operation skills were examined.Methods Magnetic resonance imaging was obtained from 9 children with mental retardation presented as OD predominantly and 11 children with mental retardation presented as LD mainly,as well as the age-matched control group (11 and 14 normal children,respectively) on a 1.5 T scanner.Voxel-based morphometry analysis with an optimization of spatial segmentation and normalization procedures was applied to compare the volume of grey matter between the two groups (OD VS.control; LD VS.control).Statistically,the total and local gray matter volumes were compared between the two groups with t test.Results The total gray matter volume of OD group was [(1.030 ± 0.078) × 106 mm3].Compared to that of controls [(0.984 ± 0.058) × 106 mm3],it was increased significantly (t =-2.6,P ＜ 0.05).And the gray matter volume in the posterior cingulated gyrus,left superior prefrontal gyrus,left cuneus,left middle prefrontal gyrus and the body of left caudate nucleus showed significantly increased.Meanwhile,the total gray matter volume of the MR children presented as LD [(1.002 ± 0.068) × 106 mm3] showed significantly increased(t =-3.0,P ＜ 0.05) compared with that of control group [(0.957 ±0.057) × 106 mm3].The gray matter volume in bilateral thalami,the left inferior temporal gyrus,the left inferior frontal gyrus,and the left cerebellum of the LD group was more than that of normal children.Conclusion As revealed by VBM,there are differences in alterations of gray matter volume between MR children presented with OD and with LD relative to control.

Objective To evaluate the clinical significance of serum levels of ProGRP, TPS and NSE in diagnosis and therapy monitoring in small cell lung cancer patients. Methods The levels of serum ProGRP, TPS and NSE in 51 SCLC patients (SCLC group), 60 benign pulmonary disease patients (benign disease group ) and 60 healthy people (healthy group ) were determined using chemiluminescent immunoassay, ELISA and electrochemiluminescent immunoassay respectively. Blood samples were collected and detected prior to therapy, before the second course of chemotherapy and the third course of chemotherapy consecutively in all the 51 SCLC patients. Results The serum ProGRP, TPS and NSE concentrations prior to chemotherapy in limited stage SCLC (LSCLC) were 136. 9(22.8-631.7)ng/L, 78. 2(56.4-114.6) U/L and 28.1(20.9-46.1)μg/L, respectively; And in extensive stage SCLC patients (ESCLC) were 1 106.6(41.2-2161.1) ng/L, 230. 9( 143.5-259.0) U/L and 81.1 (34.3-140.0)μg/L, respectively. The serum concentrations of the 3 markers in benign disease group were 19. 7 ( 9. 5-29. 1 )ng/L, 48. 7 ( 17.9-95.4) U/L and 12. 1(1.2-13.9) μg/L; and in healthy group were 20.3(10.7-30.6) ng/L, 50.3(19.5-70.7) U/L and 11.7 (1.1-13.4)μg/L, respectively. The Kruskal-Wallis test showed significantly statistical difference in different groups of the 3 tumor markers, Chi-Square were 51. 368,36. 532 and 81. 645( P ＜0. 01 ). Significant statistically differences showed when the concentrations of the 3 marks of the 2 control group were compared with that of the LSCLC group ( U =491, 827, 609 and 476, 831, 585,respectively, P ＜ 0. 05 ). Differences were also statistically significant when the 2 control group compared with that of the ESCLC group ( U = 314,532,456 and 302,553,430, respectively, P ＜ 0. 01 ). The AUC of ProGRP was 0.832 +0.029(95% CI:0.774-0.890). When cutoff value of ProGRP set as 37.7 ng/L, the diagnostic sensitivity, specificity, positive predictive value, negative predictive value and Youden's index were 71% (36/51), 97% (116/120), 90% (36/40), 89% ( 116/131 ) and 67%, respectively; show good detection performance. The sensitivity increased to 92%, 86%, 92% and 88%, when combination detection of ProGRP + TPS + NSE, ProGRP + TPS, ProGRP + NSE and TPS + NSE were used, and the specificities were 77%, 77% , 92% and 77% accordingly. The Fridman test showed significantly statistical difference in the 3 tumor markers at different stages of treatment, x2 were 49. 120, 10. 614 and 44. 392, P ＜0. 01. After the first chemotherapy course, all the tumor marker levels except TPS decreased significantly in comparison with the pretreatment concentrations. However, only ProGRP levels showed a progressive drop during the two consecutive courses of therapy, and the median concentrations were 68.0 ( 18. 6-158.4 ) and 21.0( 14. 9-63.5) ng/L (compared to the level before therapy,Z=-4. 889 and -5. 594, P ＜0. 01 ). The median of serum TPS increased slightly to 105.2 (54. 1-181.2 ) U/L after the first chemotherapy course (Z=-1.248, P＞0.05), and decreased significantly to 79.0(48.7-155.3) U/L after the second chemotherapy course (Z=-2.484, P＜0. 05 ). As to the NSE, the median concentration decreased to 11.8(8.0-16.0)μg/L after the first chemotherapy course ( Z= - 5. 568, P ＜ 0. 01 ). However, the median was 10. 6(9.0-12.7)μg/L, which showed no significant decrease after the second chemotherapy course (Z=-1.851, P＞0.05).Forty-six SCLC patients evaluated as clinical remission ( 3 CR and 43 PR) after the second chemotherapy course, among them there were 38 patients (83%) with normal serum ProGRP, TPS and NSE level ( 19 patients) or with only 1 abnormal tumor level ( 19 patients). There were only 2 patients with all abnormal serum ProGRP, TPS and NSE level, and both patients were evaluated as clinical PD. Two patients with 2 abnormal tumors results were classified as SD, the only 1 patient without therapy evaluation also had 2 abnormal tumor marker results. Conclusions The serum ProGRP, TPS and NSE are valuable tumor markers for diagnosis and treat monitoring of SCLC, particularly the ProGRP + NSE shows the highest clinical value. Combing detection of the 3 tumor markers are valuable for therapy monitoring and prognosis in SCLC patients.

Objective To detect brain structural difference between children with unexplained mental retardation and children with typically normal development. Methods The high-resolution magnetic MR imaging were obtained from 21 children with unexplained mental retardation and 30 age-matched control children without intellectual disabilities. Voxel-based morphometry analysis with an optimization of spatial segmentation and normalization procedures were applied to compare differences of gray matter volume between the two groups. The total and regional gray matter volume were compared between the two groups with independent t test. Meanwhile, correlation was conducted to analyze the relationship between the total gray matter volume and intelligence quotient (IQ) with partial correlation test. Results The total gray matter volume was significantly increased in the mental retardation children [(1. 012 ±0. 079) × 106 mm3]in relative to the controls [(0. 956 ± 0. 059) × 106 mm3, t = - 2. 80, P ＜ 0. 05]. Compared to controls,children with unexplained mental retardation showed significantly increased gray matter volume in different regions, including the bilateral thalami, the bilateral superior frontal gyri, the bilateral gyri rectus, the bilateral temporal poles, the right inferior frontal gyrus, right parahippocampal gyrus and the right cerebellum. No correlation was detected between the total gray matter volume and IQ in children with mental retardation (r = 0. 078 ,P ＞ 0. 05). Conclusions VBM would detect the gray matter abnormalities that were not founded in routine MR scanning. The increase of gray matter volume in the frontal-thalamus network might indicate the delayed maturation of the brain development. This might be one of the causations of mental retardation in children.

Objective Using gradient-echo sampling of spin-echo (GESSE) sequence to study the change of oxygen extraction fraction (OEF) in patients with unilateral cerebral vessel stenosis and the relationship between OEF and cerebral blood flow (CBF). Methods Eight normal volunteers and 16 patients with unilateral cerebral vessel stenosis were enrolled in this study. Written informed consents were obtained from all subjects. Routine MRI, GESSE and arterial spin labeling (ASL) sequences were performed for all patients. Raw data from GESSE and VE-ASL sequences were transferred to PC to conduct postprocessing. To obtain quantitative OEF and CBF of the brain parenchyma, 6 ROIs were placed respectively in the anterior, middle and posterior part of both hemispheres. The relative CBF (rCBF) was defined as the ratio of CBF of ischemic hemisphere to that of contralateral hemisphere. T test was used for statistics. Results The mean value and normal range of OEF in the volunteers were 0. 318 ± 0. 023 and 0. 272-0. 364, respectively. In the 16 patients with unilateral cerebral vessel stenosis, 8 patients had ROIs with greater OEF in unilateral hemisphere than those in contralateral hemisphere. These cases presented multiple intracranial main arterial stenoses in digital subtraction angiography (DSA) or MR angiography (MRA) examination. The other 8 patients had normal OEF in all ROIs. And they only had single arterial stenosis in DSA or MRA. Set rCBF = 0. 50 as a dividing point, the mean OEF value was 0. 397 ±0. 010 in the patients with rCBF ＜ 0. 50. In the patients with rCBF ≥ 0. 5, the mean OEF value was 0. 325 ±0. 028. The difference between the two groups was statistically significant (t = - 8. 840, P = 0. 000).Conclusion Patients with chronic cerebral ischemia may present with various hemodynamic impairment.The more CBF decreases, the more OEF increases. Those with increased OEF tended to have more than one lesion in the major intracranial arteries.

Objective To evaluate the efficacy and safety of efavirenz-based therapy in patients with human immunodeficiency virus (HIV)/hepatitis C virus (HCV) co-infection. Methods Fiftythree HIV/HCV co-infected patients received efavirenz-based therapy were followed up for 7 years.The changes of CD4+ T lymphocyte count, HIV virus load, hepatic function, hepatic fibrosis index,blood lipid, blood glucose, blood uric acid and blood routine were observed. The comparison of means before and after treatment was performed by t-test. Results The HIV RNA levels at baseline and endpoint were (4. 56±0. 88) lg copy/mL and (1.70±1.10) lg copy/mL, respectively (t=14. 781, P＜0.01). The peripheral blood CD4+ T lymphocyte counts were ( 188.37±151.14)×106/L and (445.18±314.25)×106/L, respectively (t=5.362, P＜0.01).The alanine aminotransferase (ALT) levels were (36.6±16.3) U/L and (57.2±9.9) U/L, respectively (t=7.864, P＜0. 01).The glycocholic acid levels were (444.22±476.74) mg/L and (556.88±733.05) mg/L, respectively (t=0.938, P＜0.05). The Ⅳ-collagen(Ⅳ-C) levels were (45.13±8.25) ng/mL and (47.88±4.51) ng/mL, respectively (t= 2.129, P＜0.05). The riacylglycerol levels were (1.57±0.65)mmol/L and (2.51±1.29) mmol/L, respectively (t=4.737, P＜0.01). The blood uric acid levels were (298.5±48.2) mmol/L and (495.1±89.4) mmol/L, respectively (t= 14.092, P＜0.01).Conclusions The efavirenz-based therapy is efficacious in HIV/HCV co-infected patients, but it could cause liver injury and metabolic disorder.