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 8×1012 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.

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.

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.

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>

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.

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>

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>

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>

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>

Effects of nanonetwork topography on bone marrow mesenchymal stem cell bioactivity / 中国组织工程研究

BACKGROUND:Many studies have shown that different nanostructures produce different influences on cellbioactivity, but the nanonetwork structure is not reported yet. OBJECTIVE:To study the influence of the nanonetwork topography on the bioactivity of bone marrow mesenchymal stem cels. METHODS:The nanonetwork topography was fabricated on biomedical titanium surface by alkali-heat treatment, and pure titanium served as control group. Bone marrow mesenchymal stem cels were co-cultured with the above two types of samples. cellmorphology and cytoskeleton were observed using scanning electron microscope and immunofluorescence method. The celladhesion, proliferation and osteogenic differentiation were detected by measurement of absorbance values at different culture time. RESULTS AND CONCLUSION: The nanonetwork topography had significant advantage on the number of adherent cels at 30, 60 and 120 minutes of co-culture. The cellproliferation was significantly accelerated by the nanonetwork topography at days 1, 3, 5 of co-culture, and the absorbance values in the nanonetwork group were significantly higher than those in the pure titanium group (P < 0.05). The alkaline phosphatase activity in the nanonetwork group was also significantly higher than that in the pure titanium group at 14 days of osteogenic induction (P < 0.05). The cellshape and cytoskeleton on the nanonetwork surface were better than those on the titanium surface. These findings indicate that the nanonetwork topography has better effects on cellbioactivity compared with pure titanium.

Development of articular cartilage repair technique / 中国组织工程研究

BACKGROUND:Autologous cartilage has a poor self-repair effect due to low chondrocyte density, low metabolism rate and no blood supply. OBJECTIVE:To summarize the recent study about tissue engineering techniques and surgical treatment for cartilage injury. METHODS:A computer-based online retrieval of PubMed database was performed by the first author for articles published between January 1992 and December 2013. The key words are“articular cartilage, injury, tissue engineering, repair”in English. According to the inclusion and exclusion criteria, 61 literatures were included into the final analysis. RESULTS and CONCLUSION:The current clinical treatment of articular cartilage injury includes joint debridement, mosaicplasty, perichondrium transplantation and autologous chondrocyte implantation. However, their long-term result is unsatisfactory. One reason for limited clinical success is that new cartilage can be formed at the site of a defect, and the repaired tissue canot compare with the autologous cartilage in mechanical property. Tissue engineering technique is stil a hot topic in recent years, because it can potential y induce autologous cartilage formation. Through endogenous or ectogeneous seed cells and inducting factor and nutrient factors, tissue engineering technique can be applied to induce the self-repair of articular cartilage, thus regenerating into hyalinc cartilage with the similar even same biological property. How to simplify the treatment protocols and reduce treatment cost is the key to promote cartilage repair.

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.

Transgenic mouse models of the truncated platelet integrin β3 cytoplasmic tail established by stem cell transplantation / 中国实验血液学杂志

This study was purpose to establish the transgenic mouse models of the truncated platelet integrin β3 by retrovirus-infected hematopoietic stem cells (HSCs) transplantation and to provide the basis for further study of the role of integrin β3 cytoplasmic domain in platelet bi-directional signaling pathways. Wild-type β3, β3-Δ759 (R(760) GT(762) truncated β3) and β3-Δ754 (T(755) NITYRGT(762) truncated β3) cDNAs were subcloned into MSCV MigR1 retroviral vector bearing a GFP gene and packaged into infective retrovirus with BOSC23 cell strain. The bone marrow HSCs of the β3 deficient mice were infected by the retroviruses, and transplanted into lethally-irradiated wild type C57BL/6 mice. GFP positive rate and surface β3 expression of the recipients' platelets at 6 to 8 weeks after transplantation were detected by flow cytometry to evaluate the transgenic efficiency. The results showed that four kinds of transgenic mouse models including vector, wild-type β3, β3-Δ759 and β3-Δ754 were established successfully. GFP positive rates of transgenic mouse platelets ranged from 18% to 66% and the β3 expression of transgenic mouse reached heterozygote (β3(+/-) level of mouse). It is concluded that establishment of transgenic mouse models mediated by retrovirus-infected HSCs transplantation is a feasible, fast, and high throughput transgenic approach and laid a solid foundation for further research on the role of integrin β3 cytoplasmic domain for bi-directional signaling of platelets in vivo, and for the gene therapy of platelet disorders.

A method for establishment of rat model with pulmonary contusion / 中华创伤杂志

Objective To create an easy and specific rat model of isolated bilateral pulmonary contusion and determine the maximal sublethal injury energy.Methods Injury energy was produced by free falling weights and passed through a designed precordial shield to rats' bilateral posterolateral chest wall.The rats were divided into 2.1 J,2.4 J,2.7 J and 3.0 J groups,according to the volume of injury energy.Percentage of lung contusion volume in bilateral lung was measured by blood gas analysis and three dimensional CT (3DCT) at four hours post-injury to assess lung injury severity after contusion.Pathological examination of heart and lung tissue was performed to confirm pulmonary contusion and rule out myocardial contusion.Results Death rate in 3.0 J and 2.4 J groups was 33％ and 11％,respectively.PaO2 in 2.7 J group was significantly lower than that in 2.4 J group (P ＜ 0.01),but pulmonary contusion percentage in 2.7 J group was significantly higher than that in 2.4 J group (P ＜0.01).All groups showed negative correlation between PaO2 and pulmonary contusion percentage measured by 3DCT (R2 =0.762).Hemorrhage,atelectasis and neutrophil infiltration were documented in lung biopsy.No evidence of myofiber break was recorded in heart biopsy.Conclusion This method can duplicate satisfactory models of isolated bilateral pulmonary contusion and 2.7 J can be regarded as the maximal sublethral injury energy.