OBJECTIVE To explore the levels of helper T lymphocytes(Th)in patients with hepatitis B virus(HBV)infection who were treated with entecavir and observe the impact on viral clearance.METHODS A total of 149 patients with HBV infection who were treated with entecavir in Yan'an Hospital of Kunming City from Jan.2020 to Jan.2024 were enrolled in the study,82 of whom were chronic hepatitis B(CHB),and 67 were chro-nic hepatitis B virus carriers.The enrolled patients were divided into the clearance group with 64 cases and the no clearance group with 85 cases according to the levels of serum hepatitis B surface antigen(HBsAg)at Week 72 of the treatment.The clinical data were compared between the two groups,and the changes of Th1 and Th2 levels during the treatment were analyzed.Multivariate linear regression analysis was performed for the association be-tween virological change during the treatment and immune level.The risk factors for failed clearance of viruses were analyzed by logistic regression model.RESULTS There were significant differences in the age,the levels of alanine aminotransferase(ALT)and HBV DNA between the baseline and Week 24,the levels of aspartate trans-aminase(AST)at the baseline and Week 12,the HBsAg level at Week 24,and the baseline levels of Th1,Th1/Th2 between the CHBc treatment group and the CHB treatment group(P＜0.05).There were linear correlations between the HBV DNA,HBsAg,hepatitis E antigen(HBeAg)and the Th1,Th2 and Th1/Th2,respectively(P＜0.05).Totally 64 patients were accumulatively eradicated with HBsAg on Week 72,with the eradication rate 42.95％.After the confounding factors were adjusted,multivariate analysis showed that the high levels of Th1,Th2 and Th1/Th2 were the risk factors for the failed clearance of viruses(P＜0.05).CONCLUSIONS Among the patients with HBV infection,there is difference in the immune level between the CHB patients and the CHB virus carriers.The levels of Th1,Th2 and Th1/Th2 are strongly correlated with the HBV DNA,HBsAg,HBeAg and efficiency of viral clearance during the treatment with entecavir.

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.

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 observe the characteristic symptoms in breast cancer-bearing mice and the beneficial effect of ginger-processed Jiangxiangru polysaccharides on traditional Chinese medical symptoms.Methods Thirty-eight mice were used for modeling and divided into normal,model,positive,and low-,medium-,and high-dose ginger-processed Jiangxiangru polysaccharide groups.Mice in the normal group were not inoculated with tumors and mice in the normal and model groups received physiological saline intragastrically.Mice in the positive group received celecoxib solution intragastrically,and mice in the low-,medium-,and high-dose groups received the same dose but different concentrations of ginger-processed Jiangxiangru polysaccharide solution intragastrically.Changes in body weight and tumor size were recorded after 4 weeks of continuous administration.Symptoms were observed at the end of the experiment.RGB values in photographs of the tongues,tails,and claws from mice in each group were analyzed and recorded.The degrees of blood deficiency,yin deficiency,and tumor phlegm stasis were calculated based on the method of quantitative dialectical diagnosis.The tumors were isolated and weighed,and the tumor volume and inhibition rate were calculated to determine the beneficial effect of ginger-processed Jiangxiangru polysaccharides on traditional Chinese medical symptoms.Results Mice in the breast cancer model group showed signs of blood deficiency,yin deficiency,and phlegm stasis.Tumor size was significantly reduced in mice in the ginger-processed Jiangxiangru polysaccharide groups.Ginger-processed Jiangxiangru polysaccharides inhibited tumor growth and improved blood deficiency,yin deficiency,and tumor phlegm stasis in breast cancer-bearing mice,with the best result in the high-dose group.Conclusions Ginger-processed Jiangxiangru polysaccharides can improve the symptoms of blood deficiency,yin deficiency,and tumor phlegm stasis in breast cancer-bearing mice,especially at high doses.

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 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.

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.

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.

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.

The PA-PB1 interface of the influenza polymerase is an attractive site for antiviral drug design. In this study, we designed and synthesized a mini-library of indazole-containing compounds based on rational structure-based design to target the PB1-binding interface on PA. Biological evaluation of these compounds through a viral yield reduction assay revealed that compounds 27 and 31 both had a low micromolar range of the half maximal effective concentration (EC₅₀) values against A/WSN/33 (H1N1) (8.03 μmol/L for 27; 14.6 μmol/L for 31), while the most potent candidate 24 had an EC₅₀ value of 690 nM. Compound 24 was effective against different influenza strains including a pandemic H1N1 strain and an influenza B strain. Mechanistic studies confirmed that compound 24 bound PA with a K _d which equals to 1.88 μmol/L and disrupted the binding of PB1 to PA. The compound also decreased the lung viral titre in mice. In summary, we have identified a potent anti-influenza candidate with potency comparable to existing drugs and is effective against different viral strains. The therapeutic options for influenza infection have been limited by the occurrence of antiviral resistance, owing to the high mutation rate of viral proteins targeted by available drugs. To alleviate the public health burden of this issue, novel anti-influenza drugs are desired. In this study, we present our discovery of a novel class of indazole-containing compounds which exhibited favourable potency against both influenza A and B viruses. The EC₅₀ of the most potent compounds were within low micromolar to nanomolar concentrations. Furthermore, we show that the mouse lung viral titre decreased due to treatment with compound 24. Thus our findings identify promising candidates for further development of anti-influenza drugs suitable for clinical use.