Pathogenic microorganisms are direct causative agents of zoonotic infectious diseases,po-sing severe threats to the livestock industry by inducing massive animal mortality,economic losses in livestock products,and significant risks to human health.The CRISPR/Cas system has been widely adopted in nucleic acid detection of pathogenic microorganisms due to its unique trans-cleavage activity.By leveraging the superior optical properties of nanomaterials,researchers have integrated them with CRISPR/Cas systems to develop numerous visual biosensors,which not only significantly enhance signal output but also substantially reduce detection time and cost.This re-view focuses on five nanomaterials-graphene oxide(GO),gold nanoparticles(AuNPs),MoS2 nanosheets,metal-organic frameworks(MOFs),and quantum dots(QDs)—that have been exten-sively integrated with CRISPR/Cas systems in recent years.We systematically summarize their distinct physical characteristics and specific applications in CRISPR/Cas-based pathogen detection,followed by a concise comparison of the advantages and limitations of different methodologies.Fi-nally,we discuss the prospects for nanomaterials in CRISPR/Cas detection systems,aiming to pro-vide a valuable reference for advancing molecular diagnostics of pathogenic microorganisms.

Pathogenic microorganisms are direct causative agents of zoonotic infectious diseases,po-sing severe threats to the livestock industry by inducing massive animal mortality,economic losses in livestock products,and significant risks to human health.The CRISPR/Cas system has been widely adopted in nucleic acid detection of pathogenic microorganisms due to its unique trans-cleavage activity.By leveraging the superior optical properties of nanomaterials,researchers have integrated them with CRISPR/Cas systems to develop numerous visual biosensors,which not only significantly enhance signal output but also substantially reduce detection time and cost.This re-view focuses on five nanomaterials-graphene oxide(GO),gold nanoparticles(AuNPs),MoS2 nanosheets,metal-organic frameworks(MOFs),and quantum dots(QDs)—that have been exten-sively integrated with CRISPR/Cas systems in recent years.We systematically summarize their distinct physical characteristics and specific applications in CRISPR/Cas-based pathogen detection,followed by a concise comparison of the advantages and limitations of different methodologies.Fi-nally,we discuss the prospects for nanomaterials in CRISPR/Cas detection systems,aiming to pro-vide a valuable reference for advancing molecular diagnostics of pathogenic microorganisms.

AIM:To explore the core target and mechanism of α-Linolenic acid(ALA)in improving neuroinflammation through network pharmacology combined with in vitro experiments.METHODS:Pharmacological studies have shown that ALA has anti-inflammatory,antioxidant,and neuroprotec-tive properties.The targets of α-Linolenic acid were obtained from PharmMapper and Swiss Tar-get Prediction databases,the targets of neuroin-flammation were searched from GeneCards,TTD and OMIM databases,and the potential targets of ALA and neuroinflammation were obtained from Wayne diagram.Protein interaction network(pro-tein-protein interaction,PPI)of potential targets was constructed by STRING website,and the core targets in PPI were screened by Cytoscape 3.8.0 software.At the same time,potential targets are imported into DAVID database,GO and KEGG data were obtained and the results were visualized.Autodock vina and Pymol software were used to dock the selected core targets with ALA and visual-ize the results.An in vitro model of neuroinflamma-tion was constructed,and cell growth status,oxida-tive stress,and migration or repairing capacity were determined by CCK-8 analysis,SOD,MDA and cell scratches,and the expression of IL-6,iba 1,COX-2(PTGS2),and iNOS proteins was determined by ELISA or Western blot experiments.RESULTS:Network pharmacology analysis revealed 46 poten-tial targets of ALA for neuroinflammation,and 10 core targets,including IL-6 and PTGS 2.With 232 entries enriched by GO enrichment analysis and 70 signaling pathways enriched by KEGG enrichment analysis,molecular docking showed that ALA can form hydrogen bonding with COX-2.Experiments showed that ALA could improve cell viability,allevi-ate cell oxidative stress levels,and promote cell mi-gration and motor repair in an in vitro model of neuroinflammation.CONCLUSIONS:ALA may im-prove neuroinflammation by alleviating oxidative stress and inhibiting IL-6 and COX-2 protein expres-sion.

Objective:To investigate clinical manifestations and dermoscopic characteristics of lichen planus-like keratosis (LPLK) .Methods:Clinical data were collected from 21 patients with LPLK who visited Shanghai Skin Disease Hospital and underwent both dermoscopic and histopathological examinations from January 2017 to September 2019, and clinical and dermoscopic features were retrospectively analyzed.Results:These patients were aged 64.69 ± 13.29 years, and the ratio of males to females was 1∶2. Skin lesions were located on the face of 18 cases and legs of 3 cases, and were red/violaceous in color in 7 cases, reddish-brown in 5, brown/gray in 8, and brown/reddish in 1. There were 3 types of skin lesions, including plaque-like type in 10 cases, flat pigmented patch type in 6, and flat erythema-like type in 5. As dermoscopy showed, 12 cases were non-pigmented LPLK, and 9 were pigmented LPLK. Pigment granules were found in 13 lesions, and there was no significant difference in the prevalence of pigment granules between pigmented and non-pigmented LPLK ( P=0.07) ; pigment granules were often diffusely distributed (9/13) , and the diffuse distribution pattern was common paticularly in pigmented LPLK (8/9) ; locally distributed pigment granules were found in 4 cases of non-pigmented LPLK. Coarse pigment granules were seen in 10 cases (10/13) , including 8 of pigmented LPLK and 2 of non-pigmented LPLK, and the prevalence rate of coarse pigment granules significantly differed between the pigmented LPLK and non-pigmented LPLK groups ( P=0.002) . Moreover, special distribution patterns of pigment granules included the annular granular pattern (8/13) and peppered pattern (7/13) , and no significant difference was observed in the prevalence of the 2 special distribution patterns between the pigmented LPLK and non-pigmented LPLK groups (both P > 0.05) . Scales were seen in 13 cases (13/21) , and vascular structures in 7 (7/21) , and there was no significant difference in the prevalence of the 2 structures between the pigmented and non-pigmented LPLK groups ( P=0.67, 0.16, respectively) . Conclusions:LPLK mostly occurs on the face, and manifests as solitary red, reddish-brown or brownish-gray plaques or patches, whose surfaces may be covered with scales. The characteristic dermoscopic feature of LPLK is the presence of pigment granules, which are coarse, often diffusely distributed, and commonly observed in pigmented LPLK.

Targeted programmed death-ligand 1 (PD-L1) and CXC chemokine receptor type 4 (CXCR4), gene sequences encoding anti-PD-L1 nanobody and anti-CXCR4 nanobody were cloned into the pET-22b (+) vector to construct recombinant expression plasmid of anti-PD-L1&CXCR4 bispecific nanobody, which was connected with 6 × His tag and transformed into E.coli BL21 (DE3). The expressed proteins were then found to exist as a soluble form in the supernatant of bacterial lysate after induction of IPTG.Three purification methods were used to obtain the target protein in order to improve the yield and purity of the bispecific nanobody.The bacterial supernatant was separated and purified by His Trap FF affinity chromatographic column.The target protein output could exceed 1 mg/L, and the product purity could reach up to 97%.Besides, the anti-PD-L1&CXCR4 bispecific nanobody shows a specific binding ability to two antigens on the cell surface, enhancing the cytotoxicity of IL-2 activated human peripheral blood mononuclear cells (PBMC) to tumor cell line AsPC-1, which lays the foundation for further evaluation of its drug efficacy in vivo.

Objective:To investigate the impact of diabetes mellitus (DM) and stroke on long-term outcomes in patients with acute coronary syndrome (ACS) undergoing percutaneous coronary intervention (PCI).Methods:Total 411 consecutive ACS patients undergoing PCI at the Ninth People′s Hospital of Zhengzhou between December 2014 and June 2018 were recruited, including 319 males and 92 females with a mean age of (64.7±10.1) years. These patients were divided into 4 groups according to the presence or absence of history of diabetes or stroke: non-DM non-stroke group ( n=192) , DM group ( n=140), stroke group ( n=41), and DM+stroke group ( n=38). The data of baseline demographic characteristics, clinical feature, coronary angiographic findings, and cardiovascular adverse events during long-term follow-up were obtained. Kaplan-Meier curves were used to investigate the long-term clinical outcomes among groups. Results:The mean interval of follow-up was (24.1±13.8) months. Patients with DM+stroke had the highest rates of non-fetal myocardial infarction (χ 2=24.932) , non-fetal stroke (χ 2=9.434) , hospitalization due to heart failure/angina (χ 2=69.290) , revascularization (χ 2=22.918) , cardiovascular death(χ 2=13.473)and all-cause death(χ 2=17.724)as well as hard endpoint events (the sum of non-fetal myocardial infarction, non-fetal stroke, and all-cause death) (χ 2=30.268)and combined major adverse cardiovascular events (MACE) (the sum of hard endpoint events, hospitalization due to heart failure/angina, and revascularization) (χ 2=119.556)among 4 groups(all P<0.01). In Kaplan-Meier survival analysis, the cumulative ratio of freedom from all-cause death decreased significantly in DM+stroke group compared with no DM no stroke group ( HR=17.121, 95 %CI: 2.527-115.934, P<0.01), but no statistical difference was observed in the cumulative ratio of freedom from all-cause death between DM+stroke group and DM group or stroke group respectively ( HR=3.178, 95 %CI: 0.744-13.582; HR=1.383, 95 %CI: 0.374-5.118; all P>0.05) . Meanwhile, patients with DM+stroke presented significantly lower cumulated ratio free from combined MACE than patients with non-DM non-stroke ( HR=5.423, 95 %CI:2.941-10.036, P<0.01), and the cumulated ratio free from combined MACE also decreased significantly in DM+stroke group as compared to DM group or stroke group respectively ( HR=1.859,95 %CI: 1.167-2.962; HR=1.991,95 %CI: 1.178-3.364; all P<0.01) . Conclusions:ACS patients with combined history of DM and stroke have a worse long-term outcomes after PCI than those with DM alone or stroke alone or without DM or stroke. DM and stroke seemed to have an additive effect on decrease in the cumulative ratio free from combined MACE in ACS patients following PCI.

In order to quantitatively evaluate the performance of dry electrode for fabric surface bioelectricity, a set of active measuring devices that can simulate electrocardiosignal has been developed on the basis of passive system by our group. Five Ag/AgCl fabric dry electrodes were selected to test and evaluate the devices. The results show that the deviation ratios of peak time interval of the five electrodes are all less than 1%. The maximum voltage amplitude decay rate is 7.2%, and the noise amplitudes are lower than 0.004 mV. The variable coefficient of peak time offset is less than 8%. The variable coefficient of voltage amplitude is less than 2%. The variable coefficient of noise amplitude is less than 10%. Research shows the devices has good repeatability and stability in measuring the simulated electrocardiosignal. The active measuring devices proposed in this paper can provide a new method for performance evaluation and standard formulation of surface bioelectricity dry electrode.
Electrodes ; Textiles ; Wearable Electronic Devices

Objective To observe the effect and safety of the endovascular hypothermia through hypothermic intravenous infusion device in the treatment of patients with severe craniocerebral injury compared with conventional surface cooling. Methods A total of 66 cases of patients with severe cranio-cerebral injury were randomly divided into the observation group and the control group with 33 cases in each group according to envelop randomization. The control group received surface cooling, and the observation group was given surface cooling plus endovascular hypothermia through hypothermic intravenous infusion device. The target temperature was 35 ℃maintained for 3-5 days, and natural rewarming was applied at the speed of 0.1-0.5℃/h to 36.0-37.3℃. The time to reach target temperature, the constant stability, the incidence rate and severity of complication such as shiver, arrhythmia, skin injury and agitation were recorded and compared between two groups, as well as the heart rate, breathing rate, pulse rate, blood pressure and Glasgow Coma Scale (GCS) scores after 72 h of treatment. Glasgow Outcome Scale scores 30 days after treatment and nursing workload were also calculated and compared. Results The cooling speed, time to reach target temperature and the ability to maintain at 35℃were (1.3±0.2)℃/h, (2.3±0.2) h and (6.5± 1.8)%in the observation group, respectively, compared with (0.5±0.1)℃/h, (3.6±0.6) h and (11.3±2.2)%in the control group, which had significant differences (t=1.862, 2.112, 2.408,P < 0.05). The occurrence rates of shiver, arrhythmia, skin damage and dysphoria and restlessness in the observation group were 33.33%(11/33), 9.09%(3/33), 6.06%(2/33) and 27.27%(9/33), respectively, which were much lower than those in the control group 84.85%(28/33), 15.15%(5/33), 33.33%(11/33), 54.55%(18/33),χ2=1.764-2.733,P<0.05. The heart rate, breathing rate, pulse rate, systolic blood pressure and GCS score after 72 h of treatment were(68.31 ± 3.73)times/min,(16.60 ± 1.52)times/min,(136.35 ± 3.71)mmHg(1 mmHg=0.133 kPa),(34.61 ± 1.05)℃, (9.91±4.05)points in the observation group, while(58.31±3.62)times/min,(19.81±1.83)times/min,(150.66± 2.70)mmHg,(35.65 ± 1.36)℃,(7.63 ± 3.17)points in the control group, and there were significant differences between two groups(t=2.275-3.035, P < 0.05).Besides, the ice-changing ice and turning-over time in the observation group were both remarkably reduced compared with control group, (14.03±3.11) min/h vs (38.12± 2.70) min/h (t=3.356, P<0.05) , (15.08±3.07) min/h vs (26.16±2.54) min/h ( t=3.021, P<0.05). Patients with good recovery, mild disability, severe disability, death in the observation group were 16, 13, 3 and 1 case, while 6, 11, 9, 7 cases in the control group (χ2=2.351,P < 0.05). Conclusions The endovascular hypothermia through hypothermic intravenous infusion device can rapidly reduce and effectively maintain target temperature, reduce the incidence rate of complication, improve the vital signs and decrease the nursing workload in order to improve neurological outcome in the treatment of patients with severe craniocerebral injury.

Adolescent ; Adult ; Child ; Deubiquitinating Enzyme CYLD ; Female ; Humans ; Male ; Neoplastic Syndromes, Hereditary ; genetics ; Pedigree ; Skin Neoplasms ; genetics ; Tumor Suppressor Proteins ; genetics ; Young Adult

OBJECTIVE:To observe the effect of Compound glycyrrhizin on serum VEGF of patients with alopecia areata. METHODS:36 alopecia areata patients were selected as trial group and treated with Compound glycyrrhizin(50 mg/time,3 times/d) for 3 months. Double-antibody sandwich ELISA method was adopted to determine the content of VEGF in serum before and after treatment. 36 healthy volunteers were included in control group,and serum content of VEGF in control group was compared with trial group. RESULTS:Compared with control group,the serum content of VEGF was lower in trial group,with statistical signifi-cance(t=5.863,P<0.05). After treatment,the serum VEGF levels of trial group increased,with statistical significance(t=4.351, P<0.05);there was no statistical significance in VEGF levels between trial group and control group after treatment(t=1.678,P>0.05). CONCLUSIONS:Compound glycyrrhizin can cure alopecia areata through increasing serum level of VEGF.