SPECT/CT imaging of PD-L1 expression in non-small cell lung cancer based on 99Tc m labeled anti-PD-L1 nanoantibodies / 中华核医学与分子影像杂志

Objective:To explore the value of SPECT/CT imaging on programmed death receptor 1 ligand (PD-L1) expression in patients with non-small cell lung cancer (NSCLC) based on 99Tc m labeled anti-PD-L1 nanoantibodies (NM-01). Methods:From January 2019 to March 2020, a total of 14 patients (11 males, 3 females; age: (61.9±11.0) years) with pathologically confirmed NSCLC in Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine were prospectively enrolled. NM-01 were labeled with 99Tc m, and patients were recruited for SPECT/CT imaging 2 h after injection with 99Tc m-NM-01((359.1±68.0) MBq). The differences of SUV max in primary and metastatic lesions between PD-L1 positive and negative patients were compared by independent sample t test. The correlation between the SUV max and PD-L1 expression of primary lesions was analyzed by Pearson correlation analysis. Results:Of 14 patients, 6 were PD-L1 positive and 8 were PD-L1 negative. 99Tc m-NM-01 showed obviously increased uptake in kidneys and liver, while mildly increased uptake in spleen and bone marrow. The SUV max of primary lesions was 4.69±1.88 and the SUV max of metastatic lesions was 2.04±1.32. The SUV max of primary lesions in PD-L1 positive patients was significantly higher than that of PD-L1 negative patients (5.99±1.99 vs 3.72±1.10; t=5.98, P=0.039). There was no significant difference in the SUV max of metastatic lesions between PD-L1 positive and negative patients (1.66±1.03 vs 2.35±1.46; t=-1.77, P=0.084). The SUV max of primary lesions was positively correlated with PD-L1 expression ( r=0.648, P=0.042). Conclusion:99Tc m-NM-01 can demonstrate the expression of PD-L1 in primary and metastatic lesions in NSCLC.

Epidemic situation of malaria in Yunnan Province from 2014 to 2019 / 中国血吸虫病防治杂志

Objective To investigate the epidemiological characteristics of malaria and implementation of the “1-3-7” approach in malaria elimination in Yunnan Province, so as to provide the data support for the development of post-elimination surveillance interventions. Methods All data pertaining to malaria cases in Yunnan Province from 2014 to 2019 were captured from the Notifiable Disease Reporting System of Chinese Center for Disease Control and Prevention, and the changes in the epidemic situation of malaria were analyzed during the 5-year period. In addition, the core indexes regarding the “1-3-7” approach in malaria elimination of Yunnan Province from 2014 to 2019 were retrieved from the Malaria Control System in the Parasitic Disease Information Reporting System, and all changes in the indexes were descriptively analyzed. Results During the period from 2014 to 2019, a total of 2 283 malaria cases were reported in Yunnan Province, including 1 927 cases with vivax malaria, 326 cases with plasmodium malaria, 29 cases with other species of malaria, and one case with unidentified species. There were 64 local cases, 2 219 overseas imported cases. Among the 2 283 malaria cases, the male/female ratio was 4.58∶1, and 80.25% of the cases were aged from 15 to 50 years. Farmer (70.00%) was the predominant occupation, and 76.70% (1 751/2 283) of the cases were identified in 25 border counties (districts). Malaria cases were reported in each month during the 5-year period, and the number of malaria cases increased from April, peaked on May to July, and started to decline on August. From 2014 to 2019, the reporting rate of malaria cases within 24 hours upon diagnosis was 100%, and the detection of malaria cases was 99.69% (2 276/ 2 283) in the laboratory, with a 99.65% (2 275/2 283) rate of definite diagnosis. In addition, the percentage of individual epidemiological investigations within 3 days was 100.00% (2 283/2 283), and the number of epidemic foci survey and treatment within 7 days was 576 during the 3-year period from 2017 to 2019. The goal of malaria elimination was achieved in Yunnan Province on June, 2020. Conclusions Malaria has been eliminated in Yunnan Province, and management of overseas imported malaria is the primary challenge to consolidate the malaria elimination achievements in the future. However, the approach in malaria elimination remains to be maintained, and the role of the Yunnan Provincial Malaria Diagnostic Reference Laboratory requires to be strengthened.

Improvement and application of a fast labeling system for bulk internalized vesicles / 生理学报

To study trafficking of bulk internalized vesicles such as macropinosome and lysosome in live cells, an efficient and convenient assay was established according to the axon turning assay. By injecting indicator or fluorescent dyes through a micropipette with air pressure into cell cultures to create a stable gradient around the micropipette tip, vesicles were indicated and labeled. With live cell imaging, the whole process was recorded. Without wash-out of fluorescent dyes and transferring, this assay is an effective, fast labeling system for bulk internalized vesicles, and can also be combined with imaging system.

Novel bio-mimetic receptors for early detection of Alzheimer's disease biomarkers / 中国组织工程研究

BACKGROUND: It is still a problem to achieve early and accurate diagnosis of Alzheimer's disease, and delayed treatments often occur in a large number of patients because of late diagnosis or misdiagnosis. Therefore, the development and improvement of related bioanalytical methods are of great importance for the biomarker detection of Alzheimer's disease which is still lack of means that are sensitive, efficient and low-cost. OBJECTIVE: To summarize the diagnostic methods and related biomarkers of Alzheimer's disease, and to sum up the research progress in novel biomimetic receptors for the early detection of Alzheimer's disease biomarkers. METHODS: PubMed, CNKI and Wanfang databases were used to search articles related to the biomarker studies of Alzheimer's disease and relevant studies about methods of biomarkers detection published from 2000 to 2016. The key words were "Alzheimer's disease, biomarkers, detect/detecting/detection" in Chinese and English, respectively. Finally 40 articles were obtained for the review. RESULTS AND CONCLUSION: At present, the bioanalytical methods used for biomarker detection of Alzheimer's disease mostly utilize antibodies as recognition and capture elements of biomarkers, but there are some limitations using traditional antibodies as detection receptors. Thus, novel biomimetic receptors can be substituted for conventional antibodies. Novel biomimetic receptors have high specificity, small size, low production costs and high product stability, and their chemical modification process is relatively convenient. Biomimetic receptors developed for protein analysis include aptamers, polypeptide receptors, peptoid receptors, molecularly imprinted polymers, nanobodies, gelsolin and cucurbit urils. Detection of biomarkers with novel biomimetic receptors instead of conventional antibodies will be more accurate and timely in the early diagnosis of Alzheimer's disease.

Clinical research on intelligence seven needle therapy treated infants with brain damage syndrome / 中国结合医学杂志

<p><b>OBJECTIVE</b>To assess whether the intelligence seven needle therapy administered in infants with perinatal brain damage syndrome (BDS) as early intervention would improve patients' neural development.</p><p><b>METHODS</b>A randomized controlled trial was conducted. Sixty-four infants with BDS were randomly assigned to two groups: the comprehensive group and the control group. Both groups received routine early intervention; in addition, the comprehensive group received intelligence seven needle therapy. Before and after treatment, the Bayley Scale of Infant Development (BSID), Gesell Developmental Schedules, Gross Motor Function Measure (GMFM), transcranial doppler ultrasound (TCD), and cranial imaging examination were tested for contrast.</p><p><b>RESULTS</b>After treatment, the comprehensive group showed significant difference in the Mental Development Index (MDI) scores of BSID compared with the control group (P<0.05), however, no significant discrepancy in psychomotor development index (PDI,P>0.05) was observed. The children's development quotients (DQ) of the comprehensive group exhibited a significant superiority in improving the social adaptation DQ of Gesell Developmental Schedules compared with the control group (P<0.01), as well as GMFM and linguistic and social intercourse (P<0.05). Again, no discrepancy in the fine movement DQ was found (P>0.05). The total scores of GMFM in the comprehensive group were higher than those in the control group (P<0.05). Comparing the two groups, the comprehensive group showed a significantly greater recovery rate than the control group on TCD after treatment (P<0.05). After 6-month follow-up, some recovery in both groups, specifically on broadening of brain outside space by cranial imaging examination were observed. The comprehensive group demonstrated a significantly greater recovery rate than the control group (P<0.05).</p><p><b>CONCLUSION</b>The developmental level of intelligence, motion function, linguistic competence and social intercourse can be promoted for infants with perinatal BDS by treating with the intelligence seven needle therapy. This approach can improve the brain blood supply and promote the growth of frontal and parietal lobes.</p>

Astragalus membranaceus improves endothelial-dependent vasodilator function in obese rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of Astragalus membranaceus (AM) on endothelial-dependent (EDV) and non- dependent (EIV) vascular relaxation in ex vivo thoracic aortic rings of obese rats.</p><p><b>METHODS</b>Fifteen SD rats were randomized into 3 equal groups, namely the control group fed with normal chow, obese group with high-fat chow, and AM intervention group fed with high-fat chow and daily AM gavage. The rats were sacrificed after 6 weeks of feeding, and the aortic rings were dissected and cut into 3-mm rings. The response to acethylcholine (Ach) and sodium nitroprusside (SNP) were examined in organ bath. In ex vivo study, the aortic rings obtained from the control group and obese group were incubated with AM or vehicle for 3 h in organ bath before testing the EDV and EIV. The body weight and weight of the visceral fat in each group were recorded.</p><p><b>RESULTS</b>The weight of visceral fat was greater in the obese group than in the control group, and a 6-week AM treatment significantly reduced the fat tissue due to high-fat diet. The maximum EDV value was (87.0 - or + 3.5)% in the control group, (54.8 - or + 7.8)% in the obese group, and (69.8 - or + 5.7)% in AM intervention group; the EIV values were comparable between the 3 groups. After incubation with AM, the maximum EDV values of aortic rings obtained from the obese group were significantly increased from (55.6 - or + 8.3)% to (85.1 - or + 4.5)%.</p><p><b>CONCLUSION</b>AM can improve endothelial dysfunction in obese rats, and the mechanism involves improved insulin resistance and increased endothelium-derived NO productor function.</p>

Correlation of serum levels of VEGF and SDF-1 with the number and function of circulating EPCs in children with cyanotic congenital heart disease / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To examine the number and function of circulating endothelial progenitor cells (EPCs) in children with cyanotic congenital heart diseases (CHD) and study their correlation with serum levels of vascular endothelial growth factor (VEGF) and stromal cell derived factor-1 (SDF-1).</p><p><b>METHODS</b>Fifteen children with tetralogy of Fallot (cyanotic group) and 15 age-and sex-matched children with ventricular septal defect (control group) were enrolled. Serum levels of VEGF and SDF-1 were measured using ELISA. Mononuclear cells were isolated from peripheral blood by Ficoll density gradient centrifugation and cultured in vitro. EPCs were identified by immunofluorescence and were counted under a microscope. Modified Boyden chamber assay and the MTT assay were used to measure the migration and proliferation capacities of EPCs. EPCs adhesion ability assay was performed by replating cells on fibronectin-coated dishes, and then adherent cells were counted. The correlations of serum levels of VEGF and SDF-1 with the number and function of circulating EPCs were assessed by linear regression analysis.</p><p><b>RESULTS</b>Serum levels of VEGF (201.42+/-44.74 ng/L vs 113.56+/-35.62 ng/L; P<0.05) and SDF-1 (3.45+/-1.07 ng/L vs 1.05+/-0.99 ng/L; P<0.05) in the cyanotic group were higher than those in the control group. There was a positive correlation between serum levels of VEGF and SDF-1(r=0.675, P<0.01). The number of EPCs (*200 field) in the cyanotic group significantly increased compared with that of the control group (72.2+/-9.73 vs 51.2+/-3.83; P<0.01). The functional activities of EPCs, including proliferation, migration and adhesion capacities, were augmented in the cyanotic group compared with those in the control group. The increased number and function of EPCs and the increased serum levels of VEGF and SDF-1 were consistent in the cyanotic group, with a correlation coefficient of 0.8395, 0.5491, 0.6376 and 0.7392 respectively.</p><p><b>CONCLUSIONS</b>The number and functional activity of EPCs as well as serum levels of VEGF and SDF-1 increased in children with cyanotic CHD. Serum levels of VEGF and SDF-1 were correlated to the number and functional activity of EPCs. Serum VEGF and SDF-1 together with circulating EPCs may play important roles in the pathology and physiology in these patients.</p>

Construction and expression of a prokaryotic vector of recombinant human adiponectin global domain / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct and express the recombinant human adiponectin (gAd) global domain.</p><p><b>METHODS</b>gAd complementary DNA (cDNA) was obtained from human fat tissue by RT-PCR. The PCR product was cloned into the vector pMD18-T and the prokaryotic expression vector pET32a(+). The recombinant vector was identified by digestion with double restriction endonucleases SalI and EcoRI, PCR and sequence analysis. The recombinant plasmid containing gAd gene was transformed into E. coli BL21 (DE3), and the expression of the fusion protein His-gAd was induced by IPTG.</p><p><b>RESULTS</b>The gAd cDNA of 412 bp was obtained from the total RNA of the fat tissue and verified by sequence analysis.</p><p><b>CONCLUSION</b>The recombinant plasmid could stably express the 34-kD fusion protein His-gAd in the engineered bacteria in the form of inclusion bodies.</p>

Role of apoptosis and mitochondrial apoptotic pathway in glucolipotoxicity-induced islet beta-cell dysfunction / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the mechanism of beta-cell dysfunction induced by glucolipotoxicity in high fat-fed obese rats.</p><p><b>METHODS</b>Eighteen high-fat obese male Wistar rats were assigned into 3 groups and underwent 48-hour infusion through the jugular vein with normal saline (n=6), 20% intralipid + heparin (FFA group, n=6), or 25%glucose +20% intralipid + heparin (GS-FFA group, n=6). The plasma beta-hydroxybutyric acid (beta-HBA) was measured before and at the end of the infusion. After the infusion, the rats were sacrificed following an intravenous glucose tolerance test (IVGTT) to remove the tail of the pancreas for detection of apoptotic islet cells using TUNEL method. Immunohistochemical staining was performed to detect the expression of cytochrome c (cyt c), apoptosis-inducing factor (AIF), caspase-9 and caspase-3 in the islet cells.</p><p><b>RESULTS</b>At the end of the infusion, all the rats exhibited increased plasma beta-HBA levels, which was the highest in the GS-FFA group (P<0.05). IVGTT performed after the infusion showed a significantly lower insulinogenic index in GS-FFA group than that in NS and FFA groups. Greater number of apoptotic islet cells was found in the GS-FFA group than in the FFA and NS groups (P<0.05), and the islets had significantly higher levels of cyt c, AIF, caspase-9 and caspase-3 in the former group than in the latter two groups (P<0.05).</p><p><b>CONCLUSIONS</b>Hyperglycemia and high free fatty acid level synergistically impair insulin secretions to cause ketone overproduction in high fat-fed obese rats. The beta-cell dysfunction due to glucolipotoxicity is associated with increased beta-cell apoptosis and activation of mitochondrial apoptotic pathway.</p>

Effect of decellular treatment on the framework of extracellular matrix in bovine jugular vein conduit / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of decellular treatment on the framework constituents of extracellular matrix and tissue stability in bovine jugular vein conduit (BJVC), and to provide an evidence for tissue engineering of vascular prosthesis.@*METHODS@#Bovine jugular veins were obtained fresh from a local slaughterhouse and were stored in chilled PBS. In the laboratory, any fat and loose connective tissue on the outer surface of the vessel was trimmed. BJVCs were decellularized by a 3-step extraction method as detergent Triton X-100 (0.5%), Trypsin (0.025%) EDTA (0.02%), and DNase I(30kU/L) RNaseA(0.3g/L). Histological and transmission electron microscopy (TEM) techniques were used to study the framework constituents of extracellular matrix of treated the examples, and fresh tissues were used as controls. Tissue contents of hydroxyproline(alkaline hydrolysis method) and elastin (Fastin Elastin Assay) were assayed respectively in the fresh and decellularized groups (n=10). The vascular wall heat shrinking temperature and mechanical strength were measured to evaluate the tissue stability (n=10).@*RESULTS@#Histochemical and TEM analysis of BJVCs treated with decellularization proved a complete removal of nuclear and other cell components. Tissue collagen was well kept,but elastin was partly lessened. Tissue content of hydroxyproline increased comparatively [(25.73+/-2.97)mg/g vs. (29.25+/-2.99)mg/g, P<0.05] and the elastin content obviously decreased [(159.71+/-21.06)mg/g vs. (134.91+/-35.40)mg/g, P<0.05] in the decellular treatment group compared with the control group. The heat shrinking temperature and tensile stress of decelluarized tissue were lower than those of the fresh tissue[(72.50+/-0.53) degrees C vs. (69.75+/-0.54)degrees C ,P<0.05], [(5.19+/-0.65)MPa vs. (3.13+/-0.94)MPa, P<0.05].@*CONCLUSION@#The basic framework of extracellular matrix in the decellularized BJVC is partly damaged and tissue stability is reduced. Decellularized BJVC should be further crosslinked before being used as a tissue engineering scaffold for clinical pulmonary artery graft.

Bovine jugular venous conduit treated with the polyepoxy compound / 中南大学学报(医学版)

OBJECTIVE@#To determine the feasibility whether the bovine jugular venous conduit (BJVC) can be fixed with polyepoxy compound (PC).@*METHODS@#Twenty-four BJVCs were divided into 3 groups and fixed with polyepoxy compound (PC group, n = 8), glutaraldehyde (GA group, n = 8), and unfixed group (Control group, n = 8), respectively. The morphologic and mechanical properties of BJVCs in the 3 groups, including thickness, diameter, moisture content, denaturation temperature, tensile strength, elongation at break, and fixation index were measured. The rat subcutaneous model for the assessment of tissue calcification was used. The calcium content in bovine jugular vein patches and valves was determined by flame atomic absorption spectrophotometer.@*RESULTS@#There was no difference in the wall thickness, diameter, and tissue water content between PC and the control group, but significant difference was found between GA and PC groups. The mechanical properties of PC group and GA group were not significantly different, but they were better than those of the control group. GA-fixed BJVC samples showed clear calcification, while PC fixed BJVC were calcified significantly less.@*CONCLUSION@#PC is an effective and suitable choice for the treatment of BJVC since it can effectively preserve the structure and the anti-reflow function of valves in bovine jugular vein and it has better anti-calcification properties.

The associations of HLA-DQB1 gene with onset age and autoantibodies in type 1 diabetes / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To evaluate the associations of human leukocyte antigen (HLA) DQB1 gene with onset age and autoantibodies in type 1 diabetes mellitus(T1DM) in Chinese Han population in Sichuan area.</p><p><b>METHODS</b>Forty-six type 1 diabetic patients and 52 healthy control subjects were involved in this study. HLA-DQB1 typing was performed by polymerase chain reaction-sequence specific primer(PCR-SSP). Glutamic acid decarboxylase antibody (GADA) and islet cell antibody (ICA) were qualitatively analyzed by enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The positive rate of DQB1*0201 was higher in T1DM than in controls (OR=18, P<0.005), but those of DQB1*0601, *0602 were higher in controls than in T1DM(OR=0.07, 0.31 respectively, both P<0.05).The positive rate of DQB1*0602 in type 1 diabetic patients with onset age>or=20 years was higher than that in the patients with onset age <20 years (P<0.05). GADA was more frequent in DQB1*0201(+) patients than in DQB1*0201 (-) patients (P<0.025).</p><p><b>CONCLUSION</b>The findings show that DQB1*0201 is susceptible to T1DM, whereas DQB1*0601, *0602 are protective in Chinese Han population in Sichuan area. DQB1*0602 may delay the onset of T1DM. The positive rate of DQB1*0201 correlates positively with that of GADA.</p>

Molecular scanning of candidate mtDNA gene fragment in diabetic pedigrees / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore novel pathogenic mutation in the mitochondrial DNA gene in diabetic pedigree.</p><p><b>METHODS</b>Twenty-eight suspected mitochondrial DNA diabetic families were recruited. The gene fragment was produced by PCR, and mutation was detected by direct sequencing.</p><p><b>RESULTS</b>In one pedigree, the proband and her mother were found carrying the most common nt3243 A --> G mutation and another 16S rRNA 3205C --> T mutation. But only 3205C --> T was found in her affected brother. All the two patients were deaf and developed diabetes in early age, characterized by impaired beta cell function and low body mass index (BMI). The proband had relatively higher lactic acid concentration than normal individuals. A novel ND1 gene 3434 A --> G(TAT --> TGT) mutation was explored in another proband with deafness and her affected family members.</p><p><b>CONCLUSION</b>16SrRNA 3205C --> T mutation was found in a mitochondrial diabetes mellitus pedigree, implying its potential pathogenic role in diabetes. Another novel ND1 3434 A --> G mutation was found in another diabetic pedigree. Because this mutation causes amino acid change (Tyr --> Cys) and is co-segregated with diabetes, it may be diabetogenic.</p>

Establishment of perfusion technique for isolated rat pancreas / 中华内分泌代谢杂志

Objective To establish an isolated rat pancreas perfusion technique,a method for the precise measurement of insulin secretion in vitro.Methods An isolated rat pancreas perfusion technique was applied in the study of insulin secretion from?-cells in 10 high-fat diet-induced obese Wistar rats.Results For the assessment of the functional integrity of the perfused pancreas,the isolated pancreas of 6 rats met all the criteria: (1)The constancy of perfusion pressure was kept over the whole experiment time[(70?5)mm Hg,1 mm Hg= 0.133 kPa].(2)The duodenal peristaltic activity of isolated pancreas and duodenum block was present after perfusion experiment.(3)Total insulin response to arginine stimulation was significantly increased as compared with glucose stimulation[maximum insulin secretion rate:(987?100)?U/min vs(545?50)?U/min,P