BACKGROUND:In recent years,methods for incorporating biominerals into bone tissue engineering scaffolds have been extensively studied,including solvent casting,freeze-drying,and 3D printing.However,the diverse types and complex compositions of biominerals lead to varying impacts on scaffold performance and differing requirements for fabrication processes,necessitating systematic research on their applicability.OBJECTIVE:To explore biomineral grinding and screening processes,and evaluate their solution rheological properties,hydrophilicity,mechanical properties,and biocompatibility when prepared as composite materials with polymer materials.METHODS:Five representative biominerals were selected,including turtle shell,eggshell,cuttlebone,deer antler,and pearl.These were ground into powders and screened,then mixed with polycaprolactone in specific proportions to prepare composite materials.The feasibility of applying biominerals in bone tissue engineering was explored by testing the elemental composition and particle size distribution of the powder,as well as the solution rheological properties,hydrophilicity,mechanical properties,and biocompatibility of the composite material.RESULTS AND CONCLUSION:(1)Most biomineral powders followed the rule that the longer the grinding time,the smaller the particle size.The desired particle size range could be obtained through methods such as sieving.Elemental mapping and Fourier Transform Infrared spectroscopy analyses indicated that the main inorganic mineral components of these five biominerals were calcium carbonate and calcium phosphate,containing the elements C,O,P,and Ca.(2)The method of dissolving polycaprolactone in 1,4-dioxane and adding biomineral powders to prepare scaffolds did not significantly alter the composite material composition and did not reduce biocompatibility.The addition of biomineral powders improved the hydrophilicity and 3D printability of polycaprolactone materials but decreased their mechanical properties.(3)These findings suggest that when applying biomineral powders to bone tissue engineering scaffolds,the proportion of powder added should be carefully chosen to balance hydrophilicity,printability,and mechanical properties.

BACKGROUND:In recent years,methods for incorporating biominerals into bone tissue engineering scaffolds have been extensively studied,including solvent casting,freeze-drying,and 3D printing.However,the diverse types and complex compositions of biominerals lead to varying impacts on scaffold performance and differing requirements for fabrication processes,necessitating systematic research on their applicability.OBJECTIVE:To explore biomineral grinding and screening processes,and evaluate their solution rheological properties,hydrophilicity,mechanical properties,and biocompatibility when prepared as composite materials with polymer materials.METHODS:Five representative biominerals were selected,including turtle shell,eggshell,cuttlebone,deer antler,and pearl.These were ground into powders and screened,then mixed with polycaprolactone in specific proportions to prepare composite materials.The feasibility of applying biominerals in bone tissue engineering was explored by testing the elemental composition and particle size distribution of the powder,as well as the solution rheological properties,hydrophilicity,mechanical properties,and biocompatibility of the composite material.RESULTS AND CONCLUSION:(1)Most biomineral powders followed the rule that the longer the grinding time,the smaller the particle size.The desired particle size range could be obtained through methods such as sieving.Elemental mapping and Fourier Transform Infrared spectroscopy analyses indicated that the main inorganic mineral components of these five biominerals were calcium carbonate and calcium phosphate,containing the elements C,O,P,and Ca.(2)The method of dissolving polycaprolactone in 1,4-dioxane and adding biomineral powders to prepare scaffolds did not significantly alter the composite material composition and did not reduce biocompatibility.The addition of biomineral powders improved the hydrophilicity and 3D printability of polycaprolactone materials but decreased their mechanical properties.(3)These findings suggest that when applying biomineral powders to bone tissue engineering scaffolds,the proportion of powder added should be carefully chosen to balance hydrophilicity,printability,and mechanical properties.

Percutaneous coronary intervention (PCI) is an important treatment strategy for patients with coronary artery disease. However, bleeding after PCI significantly increases the mortality risk. The search for prognostic predictors and optimal antiplatelet therapy for patients with high bleeding risk (HBR) after PCI has been a much researched upon topic in current cardiovascular research. However, there is no widely accepted prognostic model or recommended antiplatelet therapy for patients with PCI-HBR. In this trial, based on prospective multi-center database building, we will analyze the adverse prognostic predictors for patients with PCI-HBR, observe the types of antiplatelet drugs and duration of dual antiplatelet therapy in PCI-HBR patients, and compare the safety and feasibility of different antiplatelet regimens and treatment courses. The prognostic analysis and an appropriate antiplatelet strategy for patients with PCI and high bleeding risk (PPP-PCI) trial will help analyze bleeding risk factors in PCI-HBR patients and explore the appropriate antiplatelet treatment options. This study is registered with ClinicalTrials.gov (NCT05369442). The Research Ethics Committee of West China Hospital authorized this study (2022 Review #269). The trial results will be published in peer-reviewed journals and at conferences.

Percutaneous coronary intervention (PCI) is an important treatment strategy for patients with coronary artery disease. However, bleeding after PCI significantly increases the mortality risk. The search for prognostic predictors and optimal antiplatelet therapy for patients with high bleeding risk (HBR) after PCI has been a much researched upon topic in current cardiovascular research. However, there is no widely accepted prognostic model or recommended antiplatelet therapy for patients with PCI-HBR. In this trial, based on prospective multi-center database building, we will analyze the adverse prognostic predictors for patients with PCI-HBR, observe the types of antiplatelet drugs and duration of dual antiplatelet therapy in PCI-HBR patients, and compare the safety and feasibility of different antiplatelet regimens and treatment courses. The prognostic analysis and an appropriate antiplatelet strategy for patients with PCI and high bleeding risk (PPP-PCI) trial will help analyze bleeding risk factors in PCI-HBR patients and explore the appropriate antiplatelet treatment options. This study is registered with ClinicalTrials.gov (NCT05369442). The Research Ethics Committee of West China Hospital authorized this study (2022 Review #269). The trial results will be published in peer-reviewed journals and at conferences.

Aspergillus niger is an important industrial strain which has been widely used for production of enzymes and organic acids. Genome modification of A. niger is required to further improve its potential for industrial production. CRISPR/Cas9 is a widely used genome editing technique for A. niger, but its application in industrial strains modification is hampered by the need for integration of a selection marker into the genome or low gene editing efficiency. Here we report a highly efficient marker-free genome editing method for A. niger based on CRISPR/Cas9 technique. Firstly, we constructed a co-expression plasmid of sgRNA and Cas9 with a replication initiation region fragment AMA1 (autonomously maintained in Aspergillus) by using 5S rRNA promoter which improved sgRNA expression. Meanwhile, a strain deficient in non-homologous end-joining (NHEJ) was developed by knocking out the kusA gene. Finally, we took advantage of the instability of plasmid containing AMA1 fragment to cure the co-expression plasmid containing sgRNA and Cas9 through passaging on non-selective plate. With this method, the efficiency of gene editing reached 100% when using maker-free donor DNA with a short homologous arm of 20 bp. This method may facilitate investigation of gene functions and construction of cell factories for A. niger.
Gene Editing ; Aspergillus niger/genetics* ; CRISPR-Cas Systems/genetics* ; Plasmids/genetics*

【Objective】 To understand the structural characteristics of lapsed apheresis donors in our blood center and provide guidance for further improving recruitment and retention strategies by retrospectively analyzing the characteristics of lapsed apheresis donors in our center in recent years. 【Methods】 The apheresis donation and lapsed apheresis donors in Zhejiang Blood Center from 2016 to 2020 were statistically analyzed, and the general information of lapsed donors, including gender, blood type, age, education level and occupation composition, were compared and analyzed. The lapse of novel and long-term donors with different frequency were retrospectively analyzed. The SPSS 19.0 statistical software package was used for data analysis. 【Results】 In 2020, the total lapse of apheresis donors decreased by 16.6% as compared with 2016(P<0.05). The lapse rate of donors with blood groups A, B, O and AB was 1∶1∶1∶1, higher in female donors(59.0%) than males(50.0%), and dominated by age group of 18-35(66.3%). With the increase of age, the lapse rate decreased. Donors lapsed were mainly with college or above educational background(60.8%), with high proportion in students and the staff. In 2020, the lapse of novel apheresis donors decreased by 34.1% as compared with 2016, but the average lapse rate of novel apheresis donors was still as high as 70.5%. The average lapse rate of blood donors with different frequency was 52.5%, and the lapse rate decreased significantly with the increase of apheresis donation frequency. The average lapse rate of novel blood donors with whole blood donation experience was lower than those without (56.1% vs 82.9%). 【Conclusion】 Multiple measures for recruitment and retention have effectively reduced the lapse of apheresis donors. However, apheresis donors who are 18-35 years old, with college degree or above, students and staff were the main groups from lapsing. In addition, low-frequency and novel apheresis donors without whole blood donation experience were more likely to be lapsing. Targeted and personalized blood donation services should be further provided, and the management of " full cycle of blood donation" should be strengthened to reduce the lapsing, so as to retain more blood donors.

Objective To understand the radioactivity level of food surrounding a uranium mine in Xinjiang, to supplement the baseline database of food radionuclides in Xinjiang, to analyze the content of radionuclides and to estimate the internal exposure dose of residents caused by dietary intake. Methods The specific activity of ²³⁸U, ²³²Th, ²²⁶Ra, ⁴⁰K and ¹³⁷Cs of interest nuclides was measured, statistically analyzed by high purity germanium γ spectrometer, and the annual effective dose of these nuclides was estimated. Results No nuclide ²³⁸U was detected in food samples from 2018 to 2020. The average activity concentration and detection rate of other radionuclides were ²³²Th: 0.428 ± 0.038 Bq/kg (18.3%)、²²⁶Ra: 0.477 ± 0.063 Bq/kg (38%)、⁴⁰K: 162 ± 7 Bq/kg (100%)、¹³⁷Cs: 0.071 ± 0.011 Bq/kg (29.6%). The annual effective dose of residents due to food radionuclides was about 0.198 mSv. Conclusion The radioactivity level of food in the vicinity of a uranium mining mountain in Xinjiang is within the national standard limit, and the average annual effective dose caused by the diet of residents is consistent with the report of UNSCEAR in 2000. The content of artificial radionuclide ¹³⁷Cs detected in food samples will not cause harm to the health of residents.

Objective To compare the pharmacodynamics between different batches of recombinant decoy receptor innovative drug RC28-E1 and RC28-E2 in retinal angiogenesis and neovascularization,and analyze its mechanism.Methods Sixty postnatal Day 4 (P4) C57BL/6J mice were randomly divided into normal control group,vascular endothelial growth factor (VEGF)+fibroblast growth factor 2 (FGF2) group,VEGF+FGF2+RC28-E1 group,VEGF+FGF2+RC28-E2 group,VEGF+FGF2+conbercept group and VEGF+FGF2+FGF trap group by using a random number table,with 10 mice in each group.The mouse retinal explant culture system was established,and stimulated with the corresponding factors and drugs prepared in the starving culture media.The normal controls were treated with the starving media.Then the retinal explants were stained with Isolectin B4 and imaged.The number of filopodia per vascular length was quantified.In addition,ninety-six P7 C57BL/6J mice were randomly divided into normal control group,oxygen-induced retinopathy (OIR) model control group,OIR + RC28-E1 group,OIR + RC28-E2 group,OIR+conbercept group and OIR+FGF trap group by using a random number table,with 16 mice in each group.The normal controls were raised under normoxia for 10 days,and the rest of the groups were raised under hyperoxia for 5 days,then returned to normoxia for another 5 days.On P17,the retinas were isolated and stained with Isolectin B4.The stained retinas were mountedon the slides and photographed.The relative vessel obliteration and neovascularization in retina were analyzed with computer software.Then the protein levels of VEGF and FGF2 were examined by Western blot in the retinas of each group in the OIR experiment.Finally,in the RF/6A cells stimulated with VEGF and FGF2,the activities of the signaling pathways,including MEK-extracellular regulated protein kinases (Erk),protein kinase C (PKC) and protein kinase B (Akt) pathways,were examined by Western blot.All experimental procedures were evaluated and approved by the Institutional Animal Care and Use Committee of Tianjin Medical University (SYXK 2009-0001),and were in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.Results The results of retinal explant cultures showed that the numbers of filopodia per vascular length in VEGF + FGF2 + RC28-E1,VEGF + FGF2 + RC28-E2,VEGF + FGF2 + conbcrcept,and VEGF+FGF2+FGF trap groups were all significantly less than that in the VEGF+FGF2 group (all at P ＜ 0.001).The filopodia number in retinal vascular front in RC28-E1 group was similar to that in the RC28-E2 group (P =0.15),whereas the filopodia numbers in both groups were significantly decreased as compared to those in VEGF+ FGF2+conbercept group and VEGF+FGF2+FGF trap group (all at P＜0.001).The results from the OIR mouse model showed that the relative vessel obliteration area in OIR model control group was dramatically higher than those in the drug intervention groups (all at P＜0.05).There was no statistical significance in the relative vessel obliteration area between OIR+RC28-E1 group and OIR+RC28-E2 group (P =0.17),while the obliteration areas in both RC28-E-intervened groups were significantly lower than those in the OIR+conbercept group and OIR+FGF trap group (all at P＜0.05).The relative neovascular pixels in the intervention groups were significantly lower than those in the OIR model control group (all at P＜0.001).The neovascular pixels in OIR+RC28-E1 group were significantly lower than those in VEGF+FGF2+conbercept group and VEGF + FGF2 + FGF trap group (both at P ＜ 0.05),but comparable to those in OIR+RC28-E2 group (P =0.39).Western blot result showed that,the protein expression of VEGF and FGF2 in the OIR mouse retinas were significantly upregulated compared to those in the normal ones (both at P＜0.001).The upregulation of both genes were normalized by both RC28-E1 and RC28-E2.In addition,the stimulation of VEGF and FGF2 induced an enhanced activity in MEK-Erk pathway in RF/6A cells,whereas RC28-E1 inhibited the overactivation.Conclusions RC28-E1 and RC28-E2 both can inhibit angiogenesis in the retinal explants isolated from neonatal mice;they also reduce vessel obliteration and mitigate neovascularization in the OIR mouse model.Therefore,the pharmacology batch and pilot test batch of RC28-E have similar efficacies and reliable stability,and are superior in the anti-angiogenic and anti-neovascular efficacy to the currently clinically available drugs conbercept and FGF trap.RC28-E1 may suppress pathological neovascularization through inhibiting the overactivation of MEK-Erk pathway in retinal vascular endothelial cells.

Objective:To explore the application value of transcranial magnetic stimulation combined with language training for children with language retardation.Methods:100 children with language retardation who were treated in our hospital from March 2015 to March 2016 were selected as the research object.They were divided into control group and observation group according to the random number table method,50 cases in each group.The control group was treated with routine language training,while the observation group was treated with transcranial magnetic stimulation combined with language training.The treatment of the two groups were 3 months.The therapeutic effects were evaluated by the language development in Chinese children assessment method and the changes of developmental quotient before and after treatment were evaluated by the neuropsychological development test for children.Results:The effective rate of the observation group was 98.0％,which was significantly higher than 87.0％ of the control group (P＜0.05).Compared with before treatment,the language development quotient and development quotient of the two groups after treatment for 1,2 and 3 months were significantly improved,and the observation group was significantly better than the control group (P＜0.05).The normal rate of the observation group was 80.0％,which was significantly higher than 66.0％ of the control group (P＜0.05).Conclusion:The effect oftranscranial magnetic stimulation combined with language training is ideal,which can effectively improve the developmental quotient of children with language retardation,and it is worth promoting in clinical practice.

Objective To observe the effects of repetitive transcranial magnetic stimulation (rTMS) combined with rehabilitation training on motor cortex in children with motor retardation. Methods Sixty children with motor retardation were divided into treatment group and control group by random digits table method with 30 cases each. The children in control group were treated with rehabilitation training, and the children in treatment group were treated with rTMS combined with rehabilitation training. Two groups were treated for 3 courses. The Gesell pediatric neuropsychological scale and gross motor function measure (GMFM) scale before and after treatment were evaluated. Results There was no headache, dizziness and abnormal pathological reaction in the 2 groups. The motor behavior development quotient scores of Gesell pediatric neuropsychological scale after treatment in control group and treatment group were significantly higher than those before treatment: (64.37 ± 16.37) scores vs. (62.37 ± 14.21) scores and (74.50 ± 13.32) scores vs. (61.90 ± 13.76) scores, but the score in treatment was significantly higher than that in control group, and there were statistical differences (P<0.05). There was no statistical difference in language behavior development quotient score between after treatment and before treatment in control group: (79.57 ± 5.93) scores vs. (79.07 ± 5.75) scores, P>0.05. The language behavior development quotient after treatment in treatment group was significantly higher than that before treatment:(80.70 ± 5.38) scores vs. (78.57 ± 5.72) scores, and there was statistical difference (P<0.05). The A, B, C and D area scores of GMFM scale after treatment in control group and treatment group were significantly higher than before treatment, which in control group were (76.43 ± 19.18)% vs. (75.40 ± 19.38)%, (50.53 ± 27.63)%vs. (49.60 ± 28.22)%, (31.07 ± 24.93)%vs. (28.40 ± 23.70)%and (1.60 ± 1.33)%vs. (0.89 ± 0.37)%, and in treatment group were (85.80 ± 13.73)%vs. (79.13 ± 16.87)%, (65.77 ± 26.27)%vs. (49.37 ± 29.67)%, (49.60 ± 28.22)%vs. (28.83 ± 23.19)%and (2.10 ± 1.60)%vs. (1.07 ± 0.43)%, and there were statistical differences (P<0.05); the A, B and C area scores after treatment in treatment group were significantly higher than those in control group, and there were statistical differences (P<0.05); but there was no statistical difference in D area score after treatment between 2 groups (P>0.05). Conclusions The rTMS combined with rehabilitation training is better in gross movement than the single rehabilitation training for children with motor retardation. The rTMS can improve the motor function and language ability of children.