Application value of color Doppler ultrasonography combined with thyroid autoantibody detection in early diagnosis of thyroid cancer / 中国医师进修杂志

Objective:To investigate the clinical significance of color Doppler ultrasonography combined with detection of thyroid autoantibodies in the early diagnosis of thyroid cancer.Methods:A total of 108 patients with thyroid cancer who treated in Shaoxing Central Hospital Medical Community General Hospital from September 2019 to September 2021 were selected as the research group, and 108 patients with benign thyroid lesions during the same period were selected as the control group. The ultrasound examination results and the levels of serum thyroglobulin antibody (TgAb), thyroid peroxidase antibody (TPOAb) and thyroid receptor antibody (TRAb) were compared between the two groups. The relationship between the thyroid autoantibodies index and the early diagnosis of thyroid cancer were analyzed by Spearman correlation analysis; the value of early diagnosis by color Doppler ultrasonography combined with detection of thyroid autoantibodies were evaluated by the receiver operating characteristic (ROC) curve.Results:The main features of ultrasonic images in the research group were unclear edge, low echo, irregular shape, chaotic blood flow distribution, internal micro calcification, no envelope and blood flow resistance index ≥0.7. The sensitivity of ultrasonography for the diagnosis of thyroid cancer was 86.11% (93/108), the specificity was 87.18% (102/117) and the accuracy was 90.28% (195/216). The levels of serum TgAb, TPOAb and TRAb in the research group were higher than those in control group: (32.28 ± 2.85) kU/L vs. (21.96 ± 2.54) kU/L, (81.28 ± 7.32) kU/L vs. (51.53 ± 5.86) kU/L, (4.48 ± 1.25) U/L vs. (2.35 ± 0.63 ) U/L, there were statistical differences ( P<0.05). The levels of serum TgAb, TPOAb and TRAb in patients with lymph node metastasis were higher than those in the patients without lymph node metastasis: (36.28 ± 3.12) kU/L vs. (30.60 ± 2.54) kU/L, (93.51 ± 8.57) kU/L vs. (76.13 ± 6.62) kU/L, (5.73 ± 1.54) U/L vs. (3.95 ± 1.12) U/L, there were statistical differences ( P<0.05). The levels of serum TgAb, TPOAb and TRAb in patients with stage Ⅲ-Ⅳ were higher than those in the patients with stage Ⅰ-Ⅱ: (35.84 ± 3.28) kU/L vs. (29.74 ± 2.29) kU/L, (89.35 ± 8.16) kU/L vs. (75.52 ± 6.23) kU/L, (5.28 ± 1.49) U/L vs. (3.91 ± 1.25) U/L, there were statistical differences ( P<0.05). The results of Spearman correlation analysis showed that the levels of serum TgAb, TPOAb and TRAb were positively correlated with lymph node metastasis ( r = 0.758, 0.824, 0.695, P<0.05) and clinical stage of thyroid cancer ( r = 0.735, 0.796, 0.673, P<0.05). The results of ROC curve analysis showed that the area under the curve(AUC) of ultrasound examination combined with TgAb, TPOAb and TRAb for early diagnosis of thyroid cancer was 0.930, the sensitivity was 85.19%, and the specificity was 91.67%. The combined diagnostic value was better than single diagnosis. Conclusions:Ultrasound examination combined with serum TgAb, TPOAb and TRAb has high diagnostic value for early stage thyroid cancer, which is helpful to clinically clarify the condition, and provides a reliable basis for preoperative diagnosis and targeted individualized treatment plan.

Predictive value of significant coronary stenosis in patients suspected of non-ST-segment elevation acute coronary syndrome by two-dimensional speckle tracking imaging combined with real-time three-dimensional echocardiography / 中华超声影像学杂志

Objective:To evaluate the left ventricular myocardial strain and mechanical synchrony in patients suspected of non-ST-segment elevation acute coronary syndrome (NSTE-ACS) by two-dimensional speckle tracking imaging (2D-STI) and real-time three-dimensional echocardiography (RT-3DE), and to investigate the value of combined echocadiographic parameters in predication of significant coronary artery stenosis.Methods:A total of 95 patients suspected of NSTE-ACS, definitely planed to run coronary angiography (CAG) within 24-72 hours of admission were recruited in the Department of Cardiology, General Hospital of the Southern Theatre Command, PLA from December 2020 to June 2021. Regular echocardiography exam, 2D-STI and RT-3DE were performed prior to CAG.Global longitudinal peak strain (GLPS), territorial longitudinal peak strain (T RCALPS, T LADLPS, T LCXLPS) were computed by 2D-STI; the maximal difference of time to minimal systolic volume of 16-segments (Tmsv16-Dif), standard deviation of time to minimal systolic volume of 16-segment (Tmsv16-SD) and heart rate adjusted standard deviation of time to minimal systolic volume of 16-segment (Tmsv16-SD/R-R) were obtained by RT-3DE. The patients were divided into two groups according to the degree of coronary stenosis.Significant coronary artery stenosis group was defined as ≥70% of left main or any other main branch luminal narrowing ( n=53), non-significant coronary artery stenosis group was defined as <70% of luminal narrowing ( n=42). The differences of general clinical features, left ventricular strain and mechanical synchronization parameters between the two groups were compared. A binary logistic regression model was established to draw the ROC curve for predicting the severity of coronary stenosis by single and combined ultrasound parameters, and calculate the area under the ROC curve (AUC). Results:Compared with non-significant coronary artery stenosis group, GLPS were significantly reduced, while Tmsv16-SD, Tmsv16-Dif and Tmsv16-SD/R-R were significantly increased in sginificant coronary artery stenosis group (all P<0.05). The AUC of GLPS and Tmsv16-SD, Tmsv16-Dif and Tmsv16-SD/R-R for predicting significant coronary stenosis in suspected NSTE-ACS patients were 0.78, 0.69, 0.71 and 0.67, respectively. The result of joint test analysis for the dignosis of NSTE-ACS suspected significant coronary stenosis were as follows: the specificity of tandem test was 90.5%; the sensitivity of parallel test was 83.0%; the sensitivity, specificity and AUC of GLPS-Tmsv16-Dif joint index prediction test were 90.7%, 60.1% and 0.82 (95% CI=0.73-0.89) with 0.508 as Youden index. Conclusions:NSTE-ACS suspected patients with significant coronary stenosis are often accompanied by impaired left ventricular myocardial strain and mechanical dyssynchrony. A simple combination of left ventricular myocardial strain and contractility synchronization improves noninvasive prediction of high-risk coronary artery stenosis in suspected NSTE-ACS, which maybe helpful for screening patients requiring invasive examination.

Chinese expert consensus on diagnosis, treatment and prevention of venous thrombus embolism associated with chest trauma (2022 version) / 中华创伤杂志

Chest trauma is one of the most common injuries. Venous thromboembolism (VTE) as a common complication of chest trauma seriously affects the quality of patients′ life and even leads to death. Although there are some consensus and guidelines on the prevention and treatment of VTE at home and abroad, the current literatures lack specificity considering the diagnosis, treatment and prevention of VTE in patients with chest trauma have their own characteristics, especially for those with blunt trauma. Accordingly, China Chest Injury Research Society and editorial board of Chinese Journal of Traumatology organized relevant domestic experts to jointly formulate the Chinese expert consensus on the diagnosis, treatment and prevention of chest trauma venous thromboembolism associated with chest trauma (2022 version). This consensus provides expert recommendations of different levels as academic guidance in terms of the characteristics, clinical manifestations, risk assessment, diagnosis, treatment, and prevention of chest trauma-related VTE, so as to offer a reference for clinical application.

Research progress on therapeutic potential of quercetin / 中国中药杂志

Quercetin is a naturally occurring phytochemical with good bioactivity, which mainly exists in the form of glycoside in vegetables, fruits, tea, and wine and exhibits beneficial health effects. Quercetin is a dietary polyphenol that exerts the protective effects through diet or use as a food supplement. Compared with chemical agents, quercetin is widely available and safe. Quercetin has been extensively studied for its anti-diabetic, anti-hypertensive, anti-Alzheimer's disease, anti-arthritic, anti-influenza virus, anti-microbial infection, anti-aging, autophagy-regulating, and cardiovascular protective effects. Studies on its activities against different can-cer cell lines have also been reported recently. However, the poor water solubility, rapid in vivo metabolism, and short half-life of quercetin have led to its low bioavailability, thus limiting its application in the field of medicine. Quercetin nanoparticles and nanoparticle drug delivery system have been effectively utilized for enhancing its bioavailability. This paper reviewed the therapeutic potential of quercetin from both preclinical and clinical aspects and proposed solutions to improve its bioavailability, so as to provide a reference for the therapeutic application of natural compounds in the field of medicine.

Optimized AAV package and experimental application of recombinant AAV8/hFⅧ for gene therapy on hemophilia A mice / 中华血液学杂志

Objective: To evaluate the effects of adeno-associated virus (AAV) carrying hFⅧ by serotype 8 (AAV8/hFⅧ) on hemophilia A (HA) mice by gene therapy strategy. Methods: pAAV-CB-EGFP, pH22 (serotype 2) and pfΔ6 (adenovirus helper) were used to package AAV into HEK-293 cells in different conditions (ratios of cells to plasmids). The efficiency of transfection and infection were evaluated using immunofluorescence microscope to seek an optimized package condition. pAAV-TTR-hFⅧ, pH 28 (serotype 8) and pfΔ6 were applied to package AAV8/hFⅧ in HEK-293 cells using the optimized package condition. The purified AAV8/hFⅧ were intravenously injected into HA mice and the effects of gene therapy were estimated. Results: The efficiency of package was evaluated according to the amount and intensity of enhanced green fluorescent protein (EGFP) under immunofluorescence microscope. Four package conditions including 10 cm-dish to transfect 10 μg plasmids, 20 cm-dish to 20 μg, 30 μg and 40 μg plasmids were employed, and the condition of 20 cm-dish to transfect 20 μg plasmids reached the highest transfection efficiency at 24 h, 48 h and 72 h after transfection. The small scale AAV-EGFP was packaged using the optimized condition and an AAV crude extract was harvested by a freeze-thaw method. HEK-293 and 16095 cells were infected by the AAV crude extract, and the preferential infection efficiency was recognized in 16095 cells under immunofluorescence microscope. Then, AAV8/hFⅧ was packaged and purified based on the optimized transfection condition, and the high purity of AAV8/hFⅧ was detected by Western blot. Fractions of AAV8/hFⅧ at the dose of 8×10(12) vg/kg were injected into HA mice through tail vein, an eye-bleeding was performed at every two weeks, and the activity of FⅧ was measured by aPTT assay. Results showed that the activity of FⅧ maintained at the therapeutic level and lasted up to 12 weeks after injection. Conclusion: The purified AAV8/hFⅧ based on the optimized package condition could play a role in HA mice gene therapy, and the long-term therapeutic effects of AAV8/hFⅧ were observed in vivo.

Optimized AAV package and experimental application of recombinant AAV8/hFⅧ for gene therapy on hemophilia A mice / 中华血液学杂志

Objective@#To evaluate the effects of adeno-associated virus (AAV) carrying hFⅧ by serotype 8 (AAV8/hFⅧ) on hemophilia A (HA) mice by gene therapy strategy.@*Methods@#pAAV-CB-EGFP, pH22 (serotype 2) and pfΔ6 (adenovirus helper) were used to package AAV into HEK-293 cells in different conditions (ratios of cells to plasmids). The efficiency of transfection and infection were evaluated using immunofluorescence microscope to seek an optimized package condition. pAAV-TTR-hFⅧ, pH 28 (serotype 8) and pfΔ6 were applied to package AAV8/hFⅧ in HEK-293 cells using the optimized package condition. The purified AAV8/hFⅧ were intravenously injected into HA mice and the effects of gene therapy were estimated.@*Results@#The efficiency of package was evaluated according to the amount and intensity of enhanced green fluorescent protein (EGFP) under immunofluorescence microscope. Four package conditions including 10 cm-dish to transfect 10 μg plasmids, 20 cm-dish to 20 μg, 30 μg and 40 μg plasmids were employed, and the condition of 20 cm-dish to transfect 20 μg plasmids reached the highest transfection efficiency at 24 h, 48 h and 72 h after transfection. The small scale AAV-EGFP was packaged using the optimized condition and an AAV crude extract was harvested by a freeze-thaw method. HEK-293 and 16095 cells were infected by the AAV crude extract, and the preferential infection efficiency was recognized in 16095 cells under immunofluorescence microscope. Then, AAV8/hFⅧ was packaged and purified based on the optimized transfection condition, and the high purity of AAV8/hFⅧ was detected by Western blot. Fractions of AAV8/hFⅧ at the dose of 8×1012 vg/kg were injected into HA mice through tail vein, an eye-bleeding was performed at every two weeks, and the activity of FⅧ was measured by aPTT assay. Results showed that the activity of FⅧ maintained at the therapeutic level and lasted up to 12 weeks after injection.@*Conclusion@#The purified AAV8/hFⅧ based on the optimized package condition could play a role in HA mice gene therapy, and the long-term therapeutic effects of AAV8/hFⅧ were observed in vivo.

Effect of Amino Acid Motifs in Integrin β3 Cytoplasmic Tail on αⅡbβ3-Mediated Cell function in 293T cell models / 中国实验血液学杂志

OBJECTIVE@#To establish 293T cell lines stably expressing Calpain-cleavage related α3 cytoplasmic tail mutants, and to explore the effect of amino acid motifs in integrin β3 cytoplasmic tail on αⅡbβ3-mediated cell function.@*METHODS@#293T cell lines stably co-expressing human wild type integrin αⅡb and full length β3 or mutant β3, including β3-ΔNITY (β3 cytoplasmic tail NITY motif deleted), β3-Δ754 (β3 cytoplasmic tail TNITYRGT motif deleted) and β3-Δ759 (β3 cytoplasmic tail RGT motif deleted) were established. Spreading and adhesion of these stable cell lines on immobilized fibrinogen were tested.@*RESULTS@#293T-αⅡbβ3ΔNITY, 293T-αⅡbβ3Δ754, 293T-αⅡbβ3Δ759 and 293T-αⅡbβ3 cell lines were successfully established. Compared with the 293T cells, 293T-αⅡbβ3 cells which expressed full β3, possessed well adhesion and spread ability on immobilized fibrinogen, suggesting it can be as a surrogate for platelet. Compared with 293T-αⅡbβ3 cells, the 293T-αⅡbβ3ΔNITY cells showed a partial impairment of adhesion and spreadability on immobilized fibrinogen. while the 293T-αⅡbβ3Δ754 cells and 293T-αⅡbβ3Δ759 cells failed to adhere or spread on immobilized fibrinogen.@*CONCLUSION@#To the cell spreading function mediated by integrin β3, RGT motif is vital, while NITY can be dispensable. These established 293T cell lines stably expressing different β3 mutants provide a solid basis for a further analysis of mass spectrometry.

Investigation on current status of stroke care continuity in China: results and analysis of focus group interview / 中华医院管理杂志

Objective To investigate the current status and existing problems in the implementation of stroke care continuity in China, and to collect relevant suggestions for solving these problems. Methods Focus group interviews were used, to survey thirty-six nursing managers and senior nurses in neurology departments or rehabilitation programs in 24 cities of 13 provinces. The interview team members presided over the meetings, with full-time staff taking notes and recordings, and the results of the interviews were summarized and organized in a timely manner after the interview. Results At present, most of the general hospitals surveyed are doing their best to continue supporting nursing care for patients discharged from hospitals in different forms following their rescue of the acute phase. However, they are faced with such challenges as insufficient nursing manpower, and discontinuity between different medical institutions, which result in oversimplified nursing care and fragmented care. Conclusions Stroke nursing practitioners are working on continuous nursing care, with quite some challenges as well. Nursing care continuity is an important step towards establishing and improving a scientific and rational grading nursing care system.

Integrated hepatic transcriptional and serum metabolic studies on circulating nutrient metabolism in diurnal laying hens.

The aim of the study was to see the diurnal variation of nutrients metabolism and their regulation under the management of large-scaled production. The hepatic transcriptional and serum metabolic studies on circulating nutrient metabolism were investigated in diurnal laying hens. Liver and blood were collected from 36 hens that were slaughtered at 3:30, 7:30, 11:30, 15:30, 19:30, and 23:30 (n = 6), respectively. The serum amino acid, fatty acid and glucose levels, as well as the hepatic transcriptome were analyzed. The results revealed that the circadian clock genes such as Bmal1, Clock, Per1, and Cry2 displayed circadian rhythms in hen livers. The genes related to circulating nutrient transportation, lipogenesis, lipid catabolism, sterol metabolism, and oxidative/anti-oxidative systems also oscillated. However, the nadir of glucose was observed at 7:30 and peaked at 11:30 in the day. Amino acid levels peaked mainly at night, and most amino acids exhibited circadian rhythms based on CircWave analysis. With the exception of undecanoic acid (C11:0), myristoleic acid (C14:1), cis-11, 14-eicosenoic acid (C20:2), and (cis-4, 7, 10, 13, 16, 19-docosahexaenoic acid) C20:3N6 fatty acids, others peaked at 7:30 and 15:30. The results indicated that the hens required more glucose in the early morning. More proteins should be ingested late in the day, since protein catabolism occurred mostly at night. To remove the redundant fats and lipids, fewer should be ingested, especially during the night. All these results would help to design a more accurate nutrition schedule for improving the performance of laying hens in the future.

A study on the lymphocytes genetic damage of occupational population exposed to methyl tert-butyl ether / 预防医学

Objective To investigate the genetic toxicity of methyl tert-butyl ether (MTBE) to lymphocytes and to provide reference for establishing the occupational population exposure limit of MTBE. Methods The human B lymphocytes in the logarithmic growth phase were respectively exposed to MTBE at concentrations of 0, 1.25, 2.5, 5, 10 and 12.5 μmol/L. The percentage of tail DNA and Olive tail moment of human B lymphocytes were evaluated after 24 h exposure by using comet assay. Apoptosis was tested by Annexin V-FITC/PI double-staining flow cytometry. The contents of MDA and 8-OHdG, as well as GSH-Px activity were measured by ELISA kit. Sixty workers from a petrochemical factory in Zhejiang were selected as the occupational exposure population, and 55 non-occupational exposure workers were selected as the control population. And 5 mL heparin anticoagulant peripheral blood was collected, and the number of micronucleus of peripheral blood lymphocytes was measured by micronucleus test, and the percentages of tail DNA as well as Olive tail moment of peripheral blood lymphocytes were measured by comet assay. The contents of MDA and 8-OhdG and GSH-Px activity in peripheral blood plasma were measured by ELISA kit. Results After 24 h exposure, the percentage of tail DNA and Olive tail moment in human B lymphocytes at concentrations of 10-12.5 μmol/L were significantly higher than those in the control group (P<0.05), and the percentage of early apoptosis cells was significantly increased at concentrations of 10-12.5 μmol/L (P <0.01) . The results of population-based study indicated no statistically significant difference in micronucleus positive rate, and the contents of MDA and 8-OhdG, and GSH-Px activity between the exposure group and the control group, but the Olive tail moment was significantly higher in the exposure group compared with the control group (P=0.000) . Conclusion The results of vitro study showed that exposure to 10-12.5 μmol/L MTBE could cause genetic toxicity to human B lymphocytes. Olive tail moment of peripheral blood lymphocytes of occupational exposure workers was significantly higher than that of non-occupational exposure group.

Rapid determination of volatile organic compounds in workplace air by protable gas chromatography-mass spectrometer / 中华劳动卫生职业病杂志

Objective@#To establish a method for rapid determination of 47 volatile organic compounds in the air of workplace using portable gas chromatography-mass spectrometer(GC-MS).@*Methods@#The mixed standard gas with different concentration levels was made by using the static gas distribution method with the high purity nitrogen as dilution gas. The samples were injected into the GC-MS by a hand-held probe. Retention time and characteristic ion were used for qualitative analysis,and the internal standard method was usd for quantitation.@*Results@#The 47 poisonous substances were separated and determined well. The linear range of this method was 0.2-16.0 mg/m3,and the relative standard deviation of 45 volatile ovganic compounds was 3.8%-15.8%. The average recovery was 79.3%-119.0%.@*Conclusion@#The method is simple,accurate,sensitive,has good separation effect,short analysis period, can be used for qualitative and quantitative analysis of volatile organic compounds in the workplace, and also supports the rapid identification and detection of occupational hazards.

Study on Myelopoietic Functions of miR-191 by Using Zebrafish Model / 中国实验血液学杂志

<p><b>OBJECTIVES</b>To establish a method of gene analysis via zebrafish model to explore the effect of microRNA-191(miR-191) on myelopoiesis.</p><p><b>METHODS</b>The hsa-miR-191 or cel-miR-67 was microinjected into the fertilized eggs of zebrafish, then the total RNA of embryos was extracted at 10 s, 24, 36 and 48 hpf for qRT-PCR to detect the expression levels of erythroid and granulocytic genes. Then embryos at the time-point of 24 hpf were collected for whole mount in situ hybridization to detect spatiotemporal expression of those genes.</p><p><b>RESULTS</b>The antisense RNA probes with high sensitivity and specificity of erythroid genes (gata 1, scl, hbbe 3, lmo 2) and myelomonocytic genes (pu.1, L-plastin, mpx, cebpα) in zebrafish were obtained by molecular cloning and T7 RNA polymerase reaction; the expression levels of the erythroid and myelomonocytic genes of zebrafish microinjected with miR-191 mimics displayed higher than those in control group at the time-points of 24 hpf and 36 hpf. The spatiotemporal expression level of L-plastin at the time-point of 24 hpf was up-regulated, and the other genes were not significantly changed. It was worth mentioning that the mRNA expression level of mpx was significantly up-regulated by 10-20 times at the time-point of 10 s.</p><p><b>CONCLUSION</b>The genetic analysis method of embryonic myeloid differentiation has been set up via zebrafish. Preliminary analysis of regulation in zebrafish myelopoiesis shows that miR-191 may be involved in the regulation of erythroid and myelomonocytic differentiation. The mechanism and corresponding function of mpx regulated by the other factors need to be further studied.</p>

Determination of total urinary 2,5-hexanedione in the Chinese general population.

OBJECTIVE: Determination of the urinary levels of 2.5-hexanedione (2,5-HD) was performed in subjects belonging to the Chinese general population to define the reference value for this metabolite. METHODS: Urine samples were collected from 8235 individuals (4216 men and 4019 women) from the healthy general population who had not been occupationally exposed to n-hexane or methyl-n-butyl ketone. The determination was performed by a gas chromatography mass spectrometry method using an ion-trap mass spectrometer. RESULTS: The result showed that the urinary 2,5-HD median level was 0.159mg/L for the total samples. Males had statistically significant higher excretion of 2,5-HD in urine than females (median 0.171mg/L compared to 0.147mg/L, Z=-8.21, P<0.001). There was a statistically significant difference in urinary 2,5-HD levels among age groups. The excretion of 2,5-HD in urine was related to increasing age (r=-0.160, P<0.05). There was statistically significant difference in urinary 2,5-HD levels among people from difference provinces. The results showed that there was also a statistically significant effect in urinary 2,5-HD levels between current smokers and non-smokers. CONCLUSION: Finding a measurable amount of 2,5-HD in urine does not mean that the level of 2,5-HD causes an adverse health effect. Biomonitoring studies on levels of urinary 2,5-HD can provide physicians and public health officials with reference values so that they can determine whether people have been exposed to higher levels of 2,5-HD than are found in the Chinese general population. These data can also provide a foundation for scientists to make a plan for further study.

Serum and glucocorticoid kinase 1 promoted the growth and migration of non-small cell lung cancer cells.

Serum and glucocorticoid kinase 1 (SGK1) has been reported to be up-regulated in non-small cell lung cancer (NSCLC). However, its functions in NSCLC remained unclear. Here, SGK1 was found to be up-regulated in NSCLC samples. Over-expression of SGK1 promoted the growth and migration of NSCLC cells, while down-regulation of SGK1 inhibited the growth, migration and metastasis of NSCLC cells. SGK1 promoted the phosphorylation of GSK3 beta and the accumulation of beta-catenin, up-regulation of the target genes downstream of beta-catenin/TCF signaling, and activating the transcriptional activity of beta-catenin/TCF complex. Collectively, SGK1 might promote the progression of NSCLC through activating beta-catenin/TCF signaling.

Vascular endothelial injury induced by anti-endothelial cell antibody in allogeneic hematopoietic stem cell transplantation / 中华血液学杂志

<p><b>OBJECTIVE</b>To clarify the role of endothelial cells (ECs) injury induced by anti-endothelial cell antibody (AECA) in allogeneic hematopoietic stem cell transplantation (allo-HSCT).</p><p><b>METHODS</b>Serum immunoglobulin (IgG) from allo-HSCT recipients were purified and incubated with human umbilical vein vascular endothelium (HUVEC) in vitro, then the functional changes and cell apoptosis were tested.</p><p><b>RESULTS</b>After incubation with AECA positive IgG, soluble adhesion molecules significantly elevated in culture supernatant. When concentration of IgG was 160, 320, and 640 μg/ml, concentrations of soluble intercellular adhesion molecule-1 in supernatant were statistically higher in AECA positive groups [(117.10 ± 12.82) vs (78.17 ± 4.90) pg/ml, (151.30 ± 15.35) vs (89.46 ± 6.02) pg/ml, (239.00 ± 32.53) vs (127.80 ± 13.86) pg/ml, P<0.01)]. When concentration of IgG was 40, 80, 160, 320, and 640 μg/ml, concentrations of soluble vascular cell adhesion molecule-1 in supernatant were also statistically higher in AECA positive groups [(38.51 ± 3.76) vs (24.78 ± 2.59) pg/ml, (61.34 ± 6.99) vs (38.20 ± 3.17) pg/ml, (135.60 ± 24.46) vs (63.73 ± 5.08) pg/ml, (221.30 ± 29.40) vs (112.80 ± 8.91) pg/ml, (420.90 ± 31.70) vs (224.40 ± 20.79) pg/ml, P<0.01]. Clotting activity factors also elevated in culture supernatant after incubation with AECA positive IgG. When concentration of IgG was 80, 160, 320, and 640 μg/ml, concentrations of von Willebrand factor were statistically higher in AECA positive groups [(19.51 ± 0.72) vs (17.17 ± 0.60) ng/ml, P=0.0193; (22.97 ± 1.18) vs (18.27 ± 0.61) ng/ml, (26.40 ± 1.54) vs (19.53 ± 0.70) ng/ml, (34.35 ± 1.60) vs (23.81 ± 0.92) ng/ml, P<0.01]. When concentration of IgG was 320 and 640 μg/ml, concentrations of thrombomodulin were statistically higher in AECA positive groups [(57.50 ± 4.50) vs (40.31 ± 4.39) pg/ml, P=0.0132; (59.18 ± 4.11) vs (38.84 ± 5.16) pg/ml, P<0.01]. However, inflammatory factors (IL-1β, IL-6, IL-8 and ANG2) were not statistically different in AECA positive and negative groups (P>0.05). Moreover, IgG from AECA positive samples did not change the proliferation or cell apoptosis.</p><p><b>CONCLUSION</b>AECA from allo-HSCT recipients dysregulates ECs' function in vitro, but do not induce apoptosis, which is valuable in the pathophysiology of graft-versus-host disease (GVHD) and other complications after allo-HSCT.</p>

Myr-RKEFAK Peptide Selectively Regulates Outside-in Signaling Transduction-related Functions in Human Platelets / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the effect of interaction of the talin rod domain integrin binding site 2 with integrin β3 on platelet signal transduction.</p><p><b>METHODS</b>A peptide that mimics the membrane proximal α helix 6 residues R724 KEFAK729 of the integrin β3 cytoplasmic tails was designed and synthesized, to which the myristoylation was covalently linked to the N-terminal of the peptide enabling membrane penetration. The effects of myr-RKEFAK peptide on the typical platelet outside-in signaling ovent (stable adhesion and spreading on immobilized fibrinogen, aggregation, fibrin clot retraction) and inside-out signaling events (soluble fibrinogen binding) were tested.</p><p><b>RESULTS</b>myr-RKEFAK peptide dose-dependently inhibited platelet stable adhesion and spreading on immobilized fibrinogen, irreversible aggregation, as well as fibrin clot retraction, but not soluble fibrinogen binding and reversible phase of platelet aggregation.</p><p><b>CONCLUSION</b>The cell-penetrating peptide myr-RKEFAK causes an inhibitory effect on integrin β3 outside-in signaling-regulated platelets functions, but did not affect inside-out signaling-regulated platelets functions.</p>

Effect of the Integrin β3 Cytoplasmic NITY Motif on α II bβ3-Mediated Cell Functions in CHO Cell Model / 中国实验血液学杂志

<p><b>UNLABELLED</b>OBJLECTIVE: To investigate the effect of integrin β3 cytoplasmic NITY motif on αIIbβ3-mediated cell functions.</p><p><b>METHODS</b>Stable Chinese hamster ovary (CHO) cell lines that co-express human wild type integrin αIIb and wild type β3 or mutant β3ΔNITY (β3 deleting cytoplasmic NITY motif) were established. Expression of αIIb and β3 were tested by Western blot and flow cytometry in CHO cell lines. Spreading and adhesion of stable cell lines on immobilized fibrinogen were examined. The co-immunoprecipitation was used to detect protein interactions.</p><p><b>RESULTS</b>CHO-αIIbβ3, CHO-αIIbβ3ΔNITY cells were successfully established. The CHO cells transfected with wild type αIIbβ3 had the ability of adhesion and spreading. Compared with CHO-αIIbβ3 cells, CHO-αIIbβ3ΔNITY cells showed an impaired capacity of adhesion but no significant difference was observed in spreading of adhered cells. The co-immunoprecipitation showed that kindlin-2 associated with wild type integrin αIIbβ3. The β3ΔNITY mutation substantially reduced kindlin-2 association.</p><p><b>CONCLUSION</b>Deletion of NITY motif causes an impaired ability of adhesion. The deletion mutation can suppress kindlin-2 binding to integrin β3, thereby partially inhibit the integrin β3 signaling.</p>

The use of CT scan and stereo lithography apparatus technologies in a canine individualized rib prosthesis.

OBJECTIVE: To design and fabricate canine rib prosthesis with full geometric shape using computed tomography (CT) scan combined with computer-aided design (CAD) and stereo lithographic (SLA) technologies and to evaluate the accuracy of this method. METHODS: After scanned on 64 rows helical CT, the cortex part of the right 7th rib was selected as the prototype for design and manufacture of the rib prosthesis and image data were stored as DICOM format. Three-dimensional (3D) surface reconstruction was applied to produce 3D image of the 7th rib and results were outputted as STL format which were then modified by UG software for establishment of CAD model. RESULTS: The rib prosthesis with full geometric shape was obtained based on CT scanning and SLA technique. About 30,000 point cloud data were acquired after 3D laser scan of the ribs. When comparing the rib prosthesis with the rib prototype, the maximum positive deviation, maximum negative deviation, average deviation and standard deviation were 1.764 mm, -2.126 mm, 0.183/-0.253 mm and 0.346 mm, respectively. There were about 88.17% of the point cloud data within the range of ±0.5 mm. CONCLUSION: It is feasible to design and fabricate rib prosthesis with full geometric shape by using CT scanning technology combined with CAD and SLA technologies. This method is fast, convenient and precise for manufacturing prosthesis. Optimization and improvement could be processed based on the deviation suggested by the scanning.

Determination of N-Acetyl-S-(N-methylcarbamoyl) cysteine in Human Urine by Online Solid Phase Extraction-High Performance Liquid Chromatography / 分析化学

A method was developed for the determination of N-acetyl-S-( N-methylcarbamoyl ) cysteine ( AMCC) in human urine by online solid-phase extraction ( SPE )-high performance liquid chromatography ( HPLC ) . The separation of AMCC from the urine matrix was performed on AmoniPac PA Solid phase Extraction ( SPE ) column with 5 mmol/L KH2 PO4 as the mobile phase by left pump. Then the time was controlled to switch the valve to make only the section of sample containing AMCC transferred into the analytic column-Acclaim PAⅡ C18 . The determination was performed using gradient elution of 0. 1% H3 PO4 (containing 5% acetonitrile) and acetonitrile by right pump. The results showed that AMCC present good linear correlation in the range of 1 . 0-100 mg/L with a correlation coefficient of above 0 . 999 , the quantitation limit of the method was 0. 2 mg/L (with the sample inject volume =10 μL), the recoveries of spiked samples were in the range of 82 . 9%-85 . 9%, and the relative standard deviation ( n=6 ) of retention time and peak area were 0. 2% and 4. 0% respectively. Compared with offline SPE-HPLC, the proposed method was convenient, environmentally friendly, efficient and stable, and feasible for the detection of AMCC in 7 human urine samples.

Study of N-methylcarbamoylated hemoglobin in blood of being an exposure biomarker of N, N-dimethylformamide / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To assess the value of blood N-methylcarbamoylated haemoglobin (NMHb) as a biomarker of N, N-dimethylformamide (DMF) exposure.</p><p><b>METHODS</b>Seventy-two DMF processing workers in a synthetic leather factory were included in the DMF exposure group, and 12 workers in a food factory without exposure to DMF were included in the control group. Long-time individual sampling in workplace was performed among 45 workers in the exposure group, accompanied by a questionnaire survey. Blood and urine were collected for the determination of urinary N-methylformamide (NMF), urinary creatinine, and blood NMHb. Air DMF and urinary NMF were determined by gas chromatography. NMHb in blood was degraded to 3-methyl-5-isopropylhydantoin by Edman degradation before it could be determined by gas chromatography/mass spectrometry.</p><p><b>RESULTS</b>Air DMF in workplace and NMF in post-shift urine were both correlated with NMHb in blood, and the respective regression equations were LgNMHb (nmol/g globin) = 0.32×LgDMF (mg/m(3))+1.8 (r = 0.60, P < 0.005), and LgNMHb (nmol/g globin) = 0.47×LgNMF (mg/g Cr) + 1.4 (r = 0.56, P < 0.005).</p><p><b>CONCLUSION</b>NMHb can be used as a biomarker of long-term exposure to DMF.</p>