Objective:To analyze the relevant factors affecting treatment compliance in infertile patients with mycoplasma infection, construct a nomogram prediction model and validate it.Methods:In this case-control study, we retrospectively analyzed the clinical data of 400 infertile patients who attended the Department of Reproduction of Ganzhou Maternal and Child Health Hospital, Jiangxi Province from April 2022 to March 2024. They were divided into modeling group ( n=280) and validation group ( n=120) according to the 7∶3 principle with random number method, and further divided into complete compliance subgroup and unstable compliance subgroup based on different levels of compliance. Multiple logistic regression model was used to analyze the risk factors affecting treatment adherence in infertile patients with mycoplasma infection, variance inflation factor was used to assess the collinearity of the independent variables and to construct a columnar plot prediction model, and the validation and predictive efficacy of the columnar plot model were assessed by calibration curves and decision curves, respectively. The validation and predictive efficacy of the column chart model were evaluated by calibration curve and decision curve respectively. Results:In the modeling group, there was no significant statistical difference between the fully compliant group and the unstable compliant group in terms of body mass index, place of residence, occupation, settlement form, and attitude of medical staff (all P>0.05). Age in the unstable adherence subgroup [(33.22±3.20) years], proportion of junior high school and below in education [36.81% (67/182)], proportion of average monthly disposable amount ≤2 000 yuan [36.81% (67/182)], proportion of poor disease awareness [62.09% (113/182)], and infertility related stress scale (IRSS) scores (45.60±17.14) were higher than those in the full adherence subgroup [(31.36±3.24) years, P<0.001; 19.39% (19/98), P=0.003; 16.33% (16/98), P<0.001; 28.57% (28/98), P<0.001; 37.64±16.69, P<0.001]. The receiver operating characteristc curve analysis results showed that the optimal cutoff values for age and IRSS score were 32 years old and 42 points, respectively. The results of logistic multiple regression model showed that: age ( OR=1.453, 95% CI: 1.260-1.794, P=0.002), education level ( OR=1.386, 95% CI: 1.185-1.564, P<0.001), average monthly disposable amount ( OR=1.081, 95% CI: 1.002-1.246, P=0.023), disease awareness level ( OR=1.827, 95% CI: 1.390-2.359, P<0.001), and average monthly disposable amount ( OR=1.081, 95% CI: 1.002-1.246, P=0.023), disease awareness ( OR=1.827, 95% CI:1.390-2.359, P<0.001), and IRSS score ( OR=1.590, 95% CI: 1.255-1.902, P=0.006) were independent risk factors affecting treatment adherence in infertile patients with mycoplasma infection. The modeling group C-index was 0.872 (0.816-0.927), the intercept (-0.004) and slope (0.995) of the calibration curve were close to the desirable values of 0 and 1, the Brier score was 0.017, and thresholds between 0.37 and 0.96 provided a net clinical benefit. The C-index of the test model set was 0.816 (0.736-0.896), the intercept (-0.007) and slope (1.015) of the calibration curve were close to the ideal values of 0 and 1, the Brier score was 0.013, and the threshold value provided a net clinical benefit when the threshold value ranged from 0.21 to 0.98, and the H-L goodness-of-fit test showed that χ 2=7.36, P=0.499, and the constructed predictive model for the column line graphs had good discrimination and consistency. Conclusion:Age, educational level, monthly average disposable income, disease awareness, and IRSS score are the relevant factors affecting treatment compliance in infertile patients with mycoplasma infection. The nomogram prediction model constructed in this study based on those relevant factors has a good predictive value for treatment compliance in infertile patients with mycoplasma infection, and can provide certain reference for clinical intervention measures.

AIM To study the effects of total flavonoids of Dracocephalum Moldavica L.(TFDM)on reducing the inflammatory response of RAW264.7 macrophages induced by ox-LDL via the nuclear factor κB(NF-κB)/NOD-like receptor 3(NLRP3)signaling pathway.METHODS The RAW264.7 macrophages cultured in vitro were divided into the normal group,the model group(50 μg/mL ox-LDL),the TFDM group(100 μg/mL TFDM+50 μg/mL ox-LDL),the NF-κB inhibitor group(10 μmol/L Bay11-7821+50 μg/mL ox-LDL)and the TFDM+NF-κB inhibitor group(100 μg/mL TFDM+10 μmol/L Bay11-7821+50 μg/mL ox-LDL).The cells had their viability assessed by CCK-8 method;their ROS expression detected by the ROS kit;their mRNA expressions of NF-κB p65,NLRP3,Caspase-1,IL-18 and IL-1β detected by RT-qPCR;their protein expressions of NF-κB p65,IκBα,NLRP3,pro-Caspase-1,Caspase-1,IL-18 and IL-1β by Western blot;their protein expressions of NF-κB p65 and NLRP3 detected using immunofluorescence method.RESULTS Compared with the normal group,the model group showed increased ROS expression(P＜0.01);increased mRNA expressions of NF-κB p65,NLRP3,Caspase-1,IL-18 and IL-1β(P＜0.05,P＜0.01);decreased protein expressions of IκBα and cytoplasmic NF-κB p65(P＜0.01);increased protein expressions of nuclear NF-κB p65,NLRP3,Caspase-1,IL-1 β and IL-18(P＜0.01);and increased fluorescence intensity of NF-κB p65 and NLRP3(P＜0.01).Compared with the model group,the groups intervened with either TFDM or TFDM+inhibitor displayed decreased ROS expression(P＜0.01);the groups administrated with TFDM or NF-κB inhibitor,or TFDM+inhibitor showed decreased mRNA expressions of NF-κB p65,NLRP3,Caspase-1,IL-18 and IL-1β(P＜0.05,P＜0.01),increased protein expressions of IκBα and cytoplasmic NF-κB p65(P＜0.05,P＜0.01),decreased protein expressions of nuclear NF-κB p65,NLRP3,Caspase-1,IL-1β and IL-18(P＜0.05,P＜0.01),and decreased fluorescence intensity of NF-κB p65 and NLRP3(P＜0.01).There existed no significant group difference between the TFDM group and the NF-κB inhibitor group(P＞0.05).The TFDM+inhibitor group demonstrated decreased mRNA expressions of IL-1βand IL-18(P＜0.05),increased IκBα protein expression(P＜0.05),decreased protein expressions of nuclear NF-κB p65,NLRP3,Caspase-1,IL-1 β and IL-18(P＜0.05),and decreased fluorescence intensity of NLRP3 protein(P＜0.05).CONCLUSION TFDM can inhibit the ox-LDL-induced inflammatory response of RAW264.7 macrophages,and the mechansism may be associated with the reduced ROS expression and inflammatory factors due to the inhibited activation of the NF-κB/NLRP3 signaling pathway.

AIM To evaluate the quality consistency of Tongmai Granules,Tongmai Tablets,Tongmai Capsules and Tongmai Oral Liquid.METHODS The HPLC fingerprints were established,after which the contents of danshensu,protocatechuic aldehyde,3'-hydroxy puerarin,puerarin,puerarin apioside,daidzin,ferulic acid,salvianolic acid B and salvianolic acid A were determined,and cluster analysis and principal component analysis were adopted in the quality analysis from the perspective of daily intake.RESULTS There were 21 common peaks in the fingerprints for 39 batches of samples with the similarities of 0.765-0.997.Various batches of samples were clustered into 5 categories,2 principal components demonstrated the accumulative variance contribution rate of 83.53％ .The daily intakes of various constituents in different dosage forms exhibited obvious differences,especially for that of salvianolic acid B,which were low in tablets and capsules,and their heterogeneities existed among the same dosage forms.CONCLUSION This simple and accurate method can provide a reference for the quality evaluation of Tongmai preparations from different manufacturers.

Aim To identify the underlying target of bullatine A(BA)against rheumatoid arthritis(RA)u-sing limited proteolysis-small molecule mapping(LiP-SMap)drug target proteomics and to provide a scientif-ic basis for clinical application of Aconiti brachypodi Radix in the treatment of RA.Methods LiP-SMap drug target proteomics was employed to perform bioin-formatics analysis for comparing and validating the dif-ferential protein expression after BA intervention.A collagen-induced arthritis(CIA)model was estab-lished in DBA/1 mice using bovine type Ⅱ collagen.The mice were then divided into the CIA model group,methotrexate-positive control group(MTX group),and BA groups(10 mg·kg-1 and 20 mg·kg-1)based on their clinical scores.After drug intervention,the thera-peutic efficacy against RA was assessed by joint index scores and foot thickness measurements.Histopatholog-ical changes in the arthritic joints of CIA mice were e-valuated using hematoxylin and eosin(HE)staining.Enzyme-linked immunosorbent assay(ELISA)was employed to detect inflammatory cytokines interleukin-17(IL-17)and total IgG and IgG3 anti-collagen-spe-cific antibodies levels from the serum of CIA mice.Flow cytometry was used to detect the expression levels of intracellular Th17 cells(IL-17+CD4+T cells)and Th1 cells(IFN-γ+CD4+T cells).Fluorescent quanti-tative PCR was performed to detect the expression of genes related to differential proteins.Results The proteomic analysis identified Serpinb1a as a protein with strong binding affinity to BA,and KEGG enrich-ment analysis indicated IL-17 signaling pathway was a crucial pathway of BA in against RA.BA treatment significantly reduced clinical scores and foot thickness,improved local arthritis symptoms in CIA mice,and al-leviated inflammatory cell infiltration into arthritic joints(P＜0.05).Differential protein validation re-sults showed that BA had strong affinity with Serpinb1a(-5.92 kJ·mol-1)and downregulated the expres-sion of Serpinb1a mRNA.Furthermore,the administra-tion of BA markedly reduced serum IL-17 A levels from CIA mice,inhibited the expression of intracellular IL-17 A and IFN-γ cytokines in splenic CD4+T cells(P＜0.05),and significantly downregulated the transcrip-tional expression of IL-17F(P＜0.05).Conclusion BA exhibits therapeutic effects on collagen-induced arthritis,and its mechanism of action may involve the regulation of Serpinb1a and the IL-17 signaling path-way.

Objective:To analyze the relevant factors affecting treatment compliance in infertile patients with mycoplasma infection, construct a nomogram prediction model and validate it.Methods:In this case-control study, we retrospectively analyzed the clinical data of 400 infertile patients who attended the Department of Reproduction of Ganzhou Maternal and Child Health Hospital, Jiangxi Province from April 2022 to March 2024. They were divided into modeling group ( n=280) and validation group ( n=120) according to the 7∶3 principle with random number method, and further divided into complete compliance subgroup and unstable compliance subgroup based on different levels of compliance. Multiple logistic regression model was used to analyze the risk factors affecting treatment adherence in infertile patients with mycoplasma infection, variance inflation factor was used to assess the collinearity of the independent variables and to construct a columnar plot prediction model, and the validation and predictive efficacy of the columnar plot model were assessed by calibration curves and decision curves, respectively. The validation and predictive efficacy of the column chart model were evaluated by calibration curve and decision curve respectively. Results:In the modeling group, there was no significant statistical difference between the fully compliant group and the unstable compliant group in terms of body mass index, place of residence, occupation, settlement form, and attitude of medical staff (all P>0.05). Age in the unstable adherence subgroup [(33.22±3.20) years], proportion of junior high school and below in education [36.81% (67/182)], proportion of average monthly disposable amount ≤2 000 yuan [36.81% (67/182)], proportion of poor disease awareness [62.09% (113/182)], and infertility related stress scale (IRSS) scores (45.60±17.14) were higher than those in the full adherence subgroup [(31.36±3.24) years, P<0.001; 19.39% (19/98), P=0.003; 16.33% (16/98), P<0.001; 28.57% (28/98), P<0.001; 37.64±16.69, P<0.001]. The receiver operating characteristc curve analysis results showed that the optimal cutoff values for age and IRSS score were 32 years old and 42 points, respectively. The results of logistic multiple regression model showed that: age ( OR=1.453, 95% CI: 1.260-1.794, P=0.002), education level ( OR=1.386, 95% CI: 1.185-1.564, P<0.001), average monthly disposable amount ( OR=1.081, 95% CI: 1.002-1.246, P=0.023), disease awareness level ( OR=1.827, 95% CI: 1.390-2.359, P<0.001), and average monthly disposable amount ( OR=1.081, 95% CI: 1.002-1.246, P=0.023), disease awareness ( OR=1.827, 95% CI:1.390-2.359, P<0.001), and IRSS score ( OR=1.590, 95% CI: 1.255-1.902, P=0.006) were independent risk factors affecting treatment adherence in infertile patients with mycoplasma infection. The modeling group C-index was 0.872 (0.816-0.927), the intercept (-0.004) and slope (0.995) of the calibration curve were close to the desirable values of 0 and 1, the Brier score was 0.017, and thresholds between 0.37 and 0.96 provided a net clinical benefit. The C-index of the test model set was 0.816 (0.736-0.896), the intercept (-0.007) and slope (1.015) of the calibration curve were close to the ideal values of 0 and 1, the Brier score was 0.013, and the threshold value provided a net clinical benefit when the threshold value ranged from 0.21 to 0.98, and the H-L goodness-of-fit test showed that χ 2=7.36, P=0.499, and the constructed predictive model for the column line graphs had good discrimination and consistency. Conclusion:Age, educational level, monthly average disposable income, disease awareness, and IRSS score are the relevant factors affecting treatment compliance in infertile patients with mycoplasma infection. The nomogram prediction model constructed in this study based on those relevant factors has a good predictive value for treatment compliance in infertile patients with mycoplasma infection, and can provide certain reference for clinical intervention measures.

Objective To explore the brain damage of SD rats under different time points of hypobaric hypoxia exposure.Methods A rat high-altitube cerebral edema(HACE)model was constructed by simulating an altitude of 6 000 m in a hypobaric hypoxia animal experimental chamber.Thirty-six SD male rats were randomly divided into the control group and the hypobaric hypoxia exposure 3,7 and 14 d groups,with 9 rats in each group.Except for the control group,the rats in each group were continuously exposed to hypobaric hypoxia for 3,7,and 14 d.At the end of the modeling period,serum was collected by blood sampling via the abdominal aorta,and brain tissue samples were taken.The wet-to-dry ratio(W/D)of brain tissue was calculated,and the levels of relevant oxidative enzymes in serum and brain tissue were measured.The expression levels of hypoxia-inducible factor-1α(HIF-1α)and aquaporin 4(AQP4)mRNAs in brain tissue were detected by real-time fluorescence quantitative polymerase chain reaction.Results The W/D of brain tissues in the control group and the group exposed to hypobaric hypoxia for 3,7 and 14 d were 4.46±0.12,4.98±0.16,5.07±0.18 and 4.95±0.07;the superoxide dismutase contents were(111.86±2.45),(90.73±1.48),(79.64±2.56)and(55.33±1.45)U·g-1;the glutathione contents were(126.91±5.18),(125.26±1.53),(56.20±2.17)and(122.73±1.78)μg·mL-1;the malondialdehyde contents were(230.94±2.00),(362.65±3.28),(407.34±3.47)and(237.50±1.59)nmol·g-1;the relative expression levels of HIF-1 α mRNA were 1.00±0,2.99±0.49,4.72±0.49 and 1.91±0.28;the relative expression levels of AQP4 mRNA were 1.00±0,2.62±0.34,8.38±0.84 and 5.27±0.42,respectively.Statistically significant differences were found between the above indexes in the 3,7 and 14 d of hypobaric hypoxia exposure group compared with the control group(P＜0.05,P＜0.01).Conclusion Different time of hypobaric hypoxia exposure can up-regulate the expression of AQPs proteins in HACE rats and cause the disruption of the blood-brain barrier,and the HACE model constructed in the hypobaric hypoxia chamber with 6 000 m intervention for 7 d was more stable.

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.

An effective method for simultaneously detecting four semivolatile earthy-musty odors in freshwater fish by gas chromatography-mass spectrometry(GC-MS)was developed.The concurrent extraction of geosmin(GSM),2-methylisoborneol(MIB),2-isopropyl-3-methoxypyrazine(IPMP),and 2-isobutyl-3-methoxypyrazine(IBMP)in fish tissue was conducted with n-hexane.The optimized QuEChERS material was implemented,and it was found that C18,primary secondary amine(PSA)and MgSO4 could adsorb the target analytes in n-hexane.So only the graphitized carbon black(GCB)could be used to purify the extraction.The adsorption rates of different materials for the four kinds of odors materials were explored in n-hexane and ethyl acetate.The experimental results revealed that the adsorption rates of silica for the four targets were 99.5%-100%in n-hexane and 0.7%-5.0%in ethyl acetate respectively.Then the silica solid phase extraction(SPE)method was utilized to eluent the compounds using 1.0 mL n-hexane/ethyl acetate in different proportions.The results of the comparative analysis demonstrated that n-hexane/ethyl acetate(4∶1,V/V)was the optimized eluent.Based on the obtained results,n-hexane extraction and GCB purification combined with silica SPE were used to isolate GSM,MIB,IPMP and IBMP from fish and the method was validated.It was found that the method showed good linearity in the range of 0.5-200 ng/mL,and with detection limits of 0.6 μg/kg for GSM and MIB,0.2 μg/kg for IPMP and IBMP.The limits of quantitation(LOQ)were 1.0 μg/kg for GSM and MIB,0.6 μg/kg for IPMP and IBMP.Good recoveries(77.5%-112.0%)and relative standard deviations(1.56%-9.42%)were also obtained.The use of silica SPE greatly mitigated the issue that the off-flavor compounds were easily lost in the gas blowing concentration process.There was no cross contamination in this method because the sample pretreatments were conducted separately,which was different with the most commonly used HS-SPME method for detecting semi-volatile substances.The sensitivity of this method was high enough to produce good quantitative results below the odor thresholds of the examined off-flavor compounds.

OBJECTIVES: To investigate the clinical features of children with STAT gene mutations, and to explore corresponding immunotherapy strategies. METHODS: A retrospective analysis was performed for the clinical data of 10 children with STAT gene mutations who were admitted to the Department of Pediatrics of the First Affiliated Hospital of Guangzhou Medical University, from October 2015 to October 2024. Exploratory immunotherapy was implemented in some refractory cases, and the changes in symptoms, imaging manifestations, and cytokine levels were assessed after treatment. RESULTS: For the 10 children, the main clinical manifestations were recurrent rash since birth (7/10), cough (8/10), wheezing (5/10), expectoration (4/10), and purulent nasal discharge (4/10). Genotyping results showed that there was one child with heterozygous loss-of-function (LOF) mutation in the STAT1 gene, four children with heterozygous LOF mutation in the STAT3 gene, and five children with heterozygous gain-of-function (GOF) mutation in the STAT3 gene. Two children with LOF mutation in the STAT3 gene showed decreased interleukin-6 levels and improved clinical symptoms and imaging findings after omalizumab treatment. Three children with GOF mutation in the STAT3 gene achieved effective disease control after treatment with methylprednisolone (0.5 mg/kg per day). Two children with GOF mutation in the STAT3 gene received treatment with JAK inhibitor and then showed some improvement in symptoms. CONCLUSIONS STAT gene mutation screening should be considered for children with recurrent rash and purulent respiratory tract infections. Targeted immunotherapy may improve prognosis in patients with no response to conventional treatment.
Humans ; Male ; Immunotherapy ; Female ; Child, Preschool ; Child ; Gain of Function Mutation ; Retrospective Studies ; Infant ; Loss of Function Mutation ; STAT Transcription Factors/genetics*

OBJECTIVES: To investigate the application value of targeted next-generation sequencing (tNGS) in the etiological diagnosis of moderate to severe respiratory distress syndrome (RDS) in neonates. METHODS: A prospective randomized controlled trial was conducted, enrolling 81 term and late-preterm neonates with moderate to severe RDS admitted to Fujian Children's Hospital between December 2023 and December 2024. Patients were randomly assigned to the conventional microbiological test (CMT) group (n=42) or the tNGS group (n=39). For routine pathogen detection, bronchoalveolar lavage fluid was obtained via bronchoscopy, and lower respiratory tract specimens were collected via the endotracheal tube; all specimens underwent culture, and some specimens additionally underwent polymerase chain reaction or antigen testing. In the tNGS group, tNGS was performed in addition to routine pathogen detection on the same specimen types. The detection rate of pathogens, the detection rate of co-infections, and the duration of antibiotic use were compared between the two groups. RESULTS: The pathogen detection rate in the tNGS group (18/39, 46%) was significantly higher than that in the CMT group (8/42, 19%) (P=0.009). The co-infection detection rate was 13% (5/39) in the tNGS group, while no co-infections were identified in the CMT group (P=0.024). Regarding treatment, the duration of antibiotic use in the tNGS group was shorter than that in the CMT group [(12±4) days vs (15±5) days, P=0.003]. CONCLUSIONS tNGS significantly improves the pathogen detection rate in neonates with moderate to severe RDS and offers advantages in the rapid identification of co-infections and reduction of antibiotic treatment duration, suggesting it has clinical utility and potential for wider adoption.
Humans ; Prospective Studies ; Infant, Newborn ; Female ; Respiratory Distress Syndrome, Newborn/etiology* ; Male ; High-Throughput Nucleotide Sequencing/methods*