6-Thioguanine(6-TG)is an antineoplastic agent used in treatment of acute leukemia.However,significant interindividual variability in dosing regimens and frequent clinical manifestations of hepatotoxicity and myelosuppression as adverse effects have affected its therapeutic efficacy.Consequently,the development of rapid analytical methods for 6-TG in clinical samples,enabling continuous therapeutic drug monitoring of plasma concentrations,holds substantial significance in optimizing dosage regimens,mitigating adverse reactions,and investigating drug metabolism mechanisms.In this study,multi-tipped gold nanostars(AuNSs)were prepared.With bis-(p-sulfonylphenyl)phenylphosphine molecule as the protecting agent and internal standard molecule,the AuNSs were assembled onto a highly sensitive surface-enhanced Raman(SERS)substrate for developing a ratio-based SERS quantitative analysis method for 6-TG in serum.The AuNSs containing multiple tips and gaps exhibited strong local surface plasmon resonance effect and SERS activity,ensuring the sensitivity of the analytical method.Furthermore,the introduction of internal standard molecules could improve the reproducibility,which guaranteed this method suitable for rapid analysis of drug molecules in complex samples.Quantitative analysis of 6-TG was achieved with linear detetion range of 1.0×10?4-1.0 mmol/L.In the spiked recovery experiments of serum,the RSD was less than 5.32%,and the recoveries were 94%-104%,which proved that this method could be used for rapid quantitative determination of 6-TG in serum.This method provided a powerful tool for studying drug pharmacokinetics,which could promote the optimization of the usage methods of anti-cancer drugs,and it was expected to further enhance the clinical efficacy and safety of 6-TG,enabling it to achieve the best therapeutic effect.

Objective To construct a competency evaluation model for forensic practitioners,providing a reference for their training and assessment.Methods Based on the iceberg and onion models of com-petency,and with reference to Spencer's Competency Dictionary,literature research was conducted and focus group interviews were employed to preliminarily construct core indices and measurement items for evaluating the competency of forensic practitioners.The Delphi method was applied for two rounds of expert consultation to further refine the competency evaluation index system.The analytic hierarchy process(AHP)was used to calculate the weights of the indices.Results A competency evaluation model for forensic practitioners was constructed,consisting of 7 core indices,encompassing forensic skills,identification service capabilities,and the ability to apply relevant legal knowledge and 49 mea-surement items.The weights of the core indices and measurement items were determined.Conclusion The constructed competency evaluation model for forensic practitioners is scientifically sound and inno-vative,and has unique characteristics of forensic medicine compared with other medical models.

Objective To investigate the expression levels of serum miR-186-5p and miR-942-5p in patients with glioma and their clinical significance.Methods A total of 98 patients with glioma who were treated in this hospital from October 2019 to December 2021 were selected as the monitored group,and 101 healthy indi-viduals who underwent physical examinations a the same time were selected as the control group.Quantitative fluorescent PCR(qPCR)method was applied to detect the expression levels of miR-186-5p and miR-942-5p in serum,and multivariate COX regression was applied to analyze the prognostic factors of glioma patients.Re-ceiver operating characteristic(ROC)curve and area under curve(AUC)were used to analyze the diagnostic value of serum miR-186-5p and miR-942-5p in glioma.Kaplan-Meier survival curve was used to analyze the re-lationship between serum miR-186-5p and miR-942-5p expression and prognosis of patients.Results The ex-pression level of serum miR-186-5p in monitored group was lower than that in the control group(P＜0.05),and the expression level of miR-942-5p was higher than that in the control group(P＜0.05).The AUC of ser-um miR-186-5p and miR-942-5p in the diagnosis of glioma alone and in combination were 0.735,0.809 and 0.895,respectively.There were significant differences in the proportion of low miR-186-5p expression and high miR-942-5p expression in serum of patients with different preoperative Karnofsky performance status(KPS)scores,World Health Organization(WHO)grades and local infiltration(P＜0.05).The 2-year surviv-al rate of patients with high expression of miR-186-5p was higher than that of patients with low expression of miR-186-5p(x2=6.455,P=0.011).The 2-year survival rate of patients with high miR-942-5p expression was lower than that of patients with low miR-942-5p expression(x2=9.858,P=0.002).miR-186-5p was a protective factor for mortality in glioma patients(P＜0.05),while miR-942-5p was a risk factor(P＜0.05).Conclusion Serum miR-186-5p expression level decreases and miR-942-5p expression level increases in glioma patients,both of which have certain diagnostic value for the occurrence of glioma.

Immobilized enzyme-based enzyme electrode biosensors, characterized by high sensitivity and efficiency, strong specificity, and compact size, demonstrate broad application prospects in life science research, disease diagnosis and monitoring, etc. Immobilization of enzyme is a critical step in determining the performance (stability, sensitivity, and reproducibility) of the biosensors. Random immobilization (physical adsorption, covalent cross-linking, etc.) can easily bring about problems, such as decreased enzyme activity and relatively unstable immobilization. Whereas, directional immobilization utilizing amino acid residue mutation, affinity peptide fusion, or nucleotide-specific binding to restrict the orientation of the enzymes provides new possibilities to solve the problems caused by random immobilization. In this paper, the principles, advantages and disadvantages and the application progress of enzyme electrode biosensors of different directional immobilization strategies for enzyme molecular sensing elements by specific amino acids (lysine, histidine, cysteine, unnatural amino acid) with functional groups introduced based on site-specific mutation, affinity peptides (gold binding peptides, carbon binding peptides, carbohydrate binding domains) fused through genetic engineering, and specific binding between nucleotides and target enzymes (proteins) were reviewed, and the application fields, advantages and limitations of various immobilized enzyme interface characterization techniques were discussed, hoping to provide theoretical and technical guidance for the creation of high-performance enzyme sensing elements and the manufacture of enzyme electrode sensors.

To clarify the occurrence and distribution of broad bean wilt virus 2(BBWV2) on Rehmannia glutinosa, this study collected 87 R. glutinosa samples with typical symptoms of viral disease such as chlorosis and crumple from Wenxian county and Wuzhi county in Jiaozuo city, Henan province and Qiaocheng district in Bozhou city, Anhui province. The BBWV2 CP target band was amplified from 37 R. glutinosa samples by RT-PCR technology. The total detection rate reached 42.5%, among which 43.0% was detected in samples from Henan province. The detection rate in samples from Anhui province was 37.5%. 37 BBWV2 CP sequences were obtained by cloning and sequencing of BBWV2 positive samples(data has been submitted to GenBank, accession numbers: PP407959-PP407995), and the sequence analysis of these CP sequences with 91 other BBWV2 isolates in GenBank showed a high genetic diversity with a consistency rate of 70.8%-100%. Meanwhile, phylogenetic analysis showed that BBWV2 could be divided into three groups according to CP sequences, among which the BBWV2 in R. glutinosa isolates obtained in this study were all located in group 3. This study identified the differences in the occurrence, distribution, and genetic diversity of BBWV2 in R. glutinosa from Henan province and Anhui province and provided a theoretical basis for the prevention and control of BBWV2.
Rehmannia/virology* ; Phylogeny ; Plant Diseases/virology* ; China ; Molecular Sequence Data ; Fabavirus/classification*

This study aims to investigate the effect of Chaijin Jieyu Anshen Formula on the neuroinflammation of rats with insomnia complicated with depression through the regulation of triggering receptor expressed on myeloid cells 2(TREM2)/complement protein C1q signaling pathway. Rats were randomly divided into a normal group, a model group, a positive drug group, as well as a high, medium, and low-dose groups of Chaijin Jieyu Anshen Formula, with 10 rats in each group. Except for the normal group, the other groups were injected with p-chlorophenylalanine and exposed to chronic unpredictable mild stress to establish the rat model of insomnia complicated with depression. The sucrose preference experiment, open field experiment, and water maze test were performed to evaluate the depression in rats. Enzyme-linked immunosorbent assay was employed to detect serum 5-hydroxytryptamine(5-HT), dopamine(DA), and norepinephrine(NE) levels. Hematoxylin and eosin staining and Nissl staining were used to observe the damage in nucleus accumbens neurons. Western blot and immunofluorescence were performed to detect TREM2, C1q, postsynaptic density 95(PSD-95), and synaptophysin 1(SYN1) expressions in rat nucleus accumbens, respectively. Golgi-Cox staining was utilized to observe the synaptic spine density of nucleus accumbens neurons. The results show that, compared with the model group, Chaijin Jieyu Anshen Formula can significantly increase the sucrose preference as well as the distance and number of voluntary activities, shorten the immobility time in forced swimming test and the successful incubation period of positioning navigation, and prolong the stay time of space exploration in the target quadrant test. The serum 5-HT, DA, and NE contents in the model group are significantly lower than those in the normal group, with the above contents significantly increased after the intervention of Chaijin Jieyu Anshen Formula. In addition, Chaijin Jieyu Anshen Formula can alleviate pathological damages such as swelling and loose arrangement of tissue cells in the nucleus accumbens, while increasing the Nissl body numbers. Chaijin Jieyu Anshen Formula can improve synaptic damage in the nucleus accumbens and increase the synaptic spine density. Compared to the normal group, the expression of C1q protein was significantly higher in the model group, while the expression of TREM2 protein was significantly lower. Compared to the model group, the intervention with Chaijin Jieyu Anshen Formula significantly downregulated the expression of C1q protein and significantly upregulated the expression of TREM2. Compared with the model group, the PSD-95 and SYN1 fluorescence intensity is significantly increased in the groups receiving different doses of Chaijin Jieyu Anshen Formula. In summary, Chaijin Jieyu Anshen Formula can reduce the C1q protein expression, relieve the TREM2 inhibition, and promote the synapse-related proteins PSD-95 and SNY1 expression. Chaijin Jieyu Anshen Formula improves synaptic injury of the nucleus accumbens neurons, thereby treating insomnia complicated with depression.
Animals ; Male ; Rats ; Nucleus Accumbens/metabolism* ; Drugs, Chinese Herbal/administration & dosage* ; Depression/complications* ; Membrane Glycoproteins/genetics* ; Rats, Sprague-Dawley ; Sleep Initiation and Maintenance Disorders/complications* ; Neurons/metabolism* ; Receptors, Immunologic/genetics* ; Signal Transduction/drug effects* ; Synapses/metabolism*

OBJECTIVE: To investigate the clinical effect of minimally invasive technique in the treatment of moderate and severe gluteal muscle contracture. METHODS: A retrospective study was conducted on 85 patients (170 sides) with bilateral gluteal muscle contracture admitted from January 2016 to December 2019. All patients were treated with minimally invasive release of tendon knife. There were 32 males and 53 females, ranging in age from 15 to 37 years old, with an average age of (22.3±6.3) years old. Operation time, intraoperative blood loss, incision length, first postoperative ambulation time, complication rate, recurrence rate, and Harris hip score (HHS) were analyzed and evaluated. RESULTS: The average follow-up time was (16.2±4.6) months, ranging from 12 to 30 months. The operation time ranged from 7 to 15 min, with an average of (10.2±3.1) min. Intraoperative blood loss ranged from 2 to 20 ml, with an average of (8.4±2.2) ml. The incision length ranged from 0.6 to 2.0 cm, with an average of (0.8±0.3) cm. The time to postoperative ambulation ranged from 12 to 28 h, with an average of (20.0±3.2) h. All patients achieved primary wound healing without sciatic nerve injury or recurrence. HHS hip function scores ranged from 90 to 98, with an average score of (96.2±1.4). Complications included intraoperative tendon blade tip fracture in two cases (removed under fluoroscopic guidance) and subcutaneous hematoma in three cases-two resolved with compression and one with open evacuation.. Twenty-nine patients exhibited transient swaying gait postoperatively, of which 24 patients returned to normal after 4 weeks and 5 patients returned to normal after 6 weeks. CONCLUSION Minimally invasive tendon blade release is a safe and effective technique for treating gluteal muscle contracture, offering minimal trauma, rapid recovery, and excellent cosmetic and functional outcomes. However, it exhibits a low risk of blade tip fracture and sciatic nerve injury, warranting experienced surgical handling.
Humans ; Male ; Female ; Adult ; Minimally Invasive Surgical Procedures/methods* ; Adolescent ; Retrospective Studies ; Buttocks/surgery* ; Young Adult ; Contracture/surgery* ; Tendons/surgery* ; Muscle, Skeletal/surgery*

OBJECTIVES: To investigate the genomic characteristics and prognostic factors of juvenile myelomonocytic leukemia (JMML) with RAS mutations. METHODS: A retrospective analysis was conducted on the clinical data of JMML children with RAS mutations treated at the Hematology Hospital of Chinese Academy of Medical Sciences, from January 2008 to November 2022. RESULTS: A total of 34 children were included, with 17 cases (50%) having isolated NRAS mutations, 9 cases (27%) having isolated KRAS mutations, and 8 cases (24%) having compound mutations. Compared to children with isolated NRAS mutations, those with NRAS compound mutations showed statistically significant differences in age at onset, platelet count, and fetal hemoglobin proportion (P<0.05). Cox proportional hazards regression model analysis revealed that hematopoietic stem cell transplantation (HSCT) and hepatomegaly (≥2 cm below the costal margin) were factors affecting the survival rate of JMML children with RAS mutations (P<0.05); hepatomegaly was a factor affecting survival in the non-HSCT group (P<0.05). CONCLUSIONS Children with NRAS compound mutations have a later onset age compared to those with isolated NRAS mutations. At initial diagnosis, children with NRAS compound mutations have poorer peripheral platelet and fetal hemoglobin levels than those with isolated NRAS mutations. Liver size at initial diagnosis is related to the prognosis of JMML children with RAS mutations. HSCT can improve the prognosis of JMML children with RAS mutations.
Humans ; Leukemia, Myelomonocytic, Juvenile/therapy* ; Mutation ; Male ; Female ; Child, Preschool ; Retrospective Studies ; Child ; Infant ; GTP Phosphohydrolases/genetics* ; Membrane Proteins/genetics* ; Adolescent ; Hematopoietic Stem Cell Transplantation ; Proportional Hazards Models ; Proto-Oncogene Proteins p21(ras)/genetics* ; Prognosis

Aim To investigate the effect of KHSRP on the malignant biological behavior of gastric cardia ade-nocarcinoma by targeting JAK1/STAT3 signaling axis.Methods The expression levels of KHSRP in adeno-carcinoma tissues of gastric cardia adenocarcinoma and adjacent tissues were collected and compared.qRT-PCR experiment to detect transfection efficiency in gas-tric cardia adenocarcinoma cell lines(OE-19,TE-7,BIC-1,FLO-1,SK-GT-4,BE-3)and normal gastric mucosal epithelial cell line(GES-1).The knockdown and overexpression of KHSRP were treated by packa-ging lentiviral Vector,and the cells were divided into sh-NC group,sh-KHSRP group,vector group,and KH-SRP group.Cell counting kit-8(CCK-8)and Tran-swell assay were used to determine the effects of KH-SRP on the proliferation,migration and invasion of ade-nocarcinoma cells of gastric cardia adenocarcinoma.Xenograft tumor models were used to detect the effects of knockdown and overexpression of KHSRP in live an-imals.WB experiments confirmed that KHSRP targeted the JAK/STAT signaling pathway.Results KHSRP was overexpressed in GCA tissues and cell lines(P＜0.05).Cell function assay analysis showed that KH-SRP overexpression significantly promoted GCA cell proliferation,migration and invasion in vitro(P＜0.05).After KHSRP knockdown,the phosphorylation levels of JAK1 and STAT3 in JAK/STAT signaling pathway were significantly decreased,and the situation was opposite after KHSRP overexpression(P＜0.05).Conclusion KHSRP regulates the malignant progres-sion of metastasis of gastric cardia adenocarcinoma by activating JAK1/STAT3 signaling axis.

Objective:To investigate the immuno-inflammatory regulatory effects of the Qing Gan San Jie Xiao Ying Formula(QGSJXYF)on Hashimoto's thyroiditis(HT)by modulating thyroid cell-derived exosomes to provide experi-mental evidence for its immunomodulatory mechanisms.Methods:Nthy-ori-3-1 thyroid cells were treated with QGSJXYF-medicated serum,with untreated cells serving as controls.Exosomes from both groups were extracted and analyzed using nanoparticle tracking analysis(NTA),transmission electron microscopy(TEM),and Western blot to assess concentration,size distribution,morphology,and the expression of characteristic exosomal markers.An inflammatory model of human T lymphocytes(H9)was established and co-incubated with normal exosomes(EXO-C group)or QGSJXYF-treated exosomes(EXO-T group).The levels of inflammatory cytokines in H9 cells were measured using Western blot(WB)and ELISA.Results:Exosome characterization showed that the particle concentration of Nthy-ori-3-1 cell-derived exosomes in both the control and QGSJXYF groups ranged from 1×109 to 1×1011/mL,with particle diameters between 80～300 nm.The exosomes exhibited a typical spherical or cup-shaped morphology with positive expression of TSG101,CD63,and HSP70.Compared with the inflammation model group and the EXO-C group,the EXO-T group significantly reduced the intracellular expression of IL-17A protein in H9 cells(P<0.05)and suppressed IL-17 and IL-6 levels in the cell supernatant(P<0.01).Conclusion:QGSJXYF may exert its anti-inflammatory and thyroid-protective effects by modulating the functional state of thyroid cell-derived exosomes,regulating the inflamma-tory microenvironment,and inhibiting the expression of inflammatory cytokines associated with Hashimoto's thyroiditis.