ObjectiveTo study the effects of applying different microbial fertilizers on the growth and rhizosphere soil environment of Codonopsis pilosula and provide a theoretical basis for ecological cultivation of this medicinal plant. MethodsSeven groups were designed， including CK （no application of microbial fertilizer）， T1 （Trichoderma longibrachiatum fertilizer）， T2 （Bacillus subtilis fertilizer）， T3 （Trichoderma viride fertilizer）， T4 （compound microbial fertilizer）， T5 （C. pilosula stems and leaves fermented with compound microbial fertilizer）， and T6 （Scutellaria baicalensis stems and leaves fermented with T. viride fertilizer）. The physiological indicators， yield， and quality of C. pilosula and the physicochemical properties， enzyme activities， and microbial diversity in the rhizosphere soil of different fertilizer treatments were measured. ResultsGroup T1 showed slight decreases in soluble protein content （SPC） and superoxide dismutase （SOD）. Groups T2-T6 showed increases in physiological indicators such as proline （Pro）， soluble solids content （SSC）， SPC， catalase （CAT）， and peroxidase （POD） and a decrease in malondialdehyde （MDA） in C. pilosula leaves. All the fertilizer treatments increased the yield of C. pilosula and the total polysaccharide content in the roots. T1， T2， T3， T4， and T5 increased the total flavonoid content in the roots. Meanwhile， T4 increased the total saponin content in the roots. All the fertilizer treatments reduced the pH and increased the electric conductivity （EC）， soil organic matter （SOM）， and alkaline nitrogen （AN） in the soil. T2 and T5 increased the available phosphorus （AP）， and T3， T4， T5， and T6 increased the available potassium （AK） in the soil. All the fertilizer treatments increased the activities of urease， sucrase， and CAT in the soil. Except that T1 decreased the bacterial diversity in the soil， other fertilizer treatments significantly increased bacterial and fungal diversity in the soil. Different fertilizer treatments significantly affected the composition of bacterial and fungal communities in the soil. At the phylum level， the dominant bacterial phyla included Proteobacteria， Acidobacteriota， and Bacteroideta， and the dominant fungal phyla were Ascomycota， Mortierellomycota， and unclassified_fungi in the rhizosphere soil of C. pilosula after bacterial fertilizer treatment. At the genus level， unclassified Gemmatimonadaceae， Sphingomonas， and unclassified Vicinamibacteraceae were the dominant bacterial genera， while unidentified， unclassified Fungi， and unclassified Sordariomycetes were the dominant fungal genera in the rhizosphere soil. The results of redundancy analysis indicated that the main physicochemical factors affecting changes of microbial communities in the rhizosphere soil of C. pilosula were pH， EC， AK， AN， AP， and soil organic matter （SOM） in the soil. The correlation heatmap showed that Bryobacter had significantly positive correlations with EC， AK， and AN. There was a significantly negative correlation between Fusarium and SOM. In summary， applying an appropriate amount of microbial fertilizer can promote the growth and improve the rhizosphere soil environment of C. pilosula. ConclusionThe compound microbial fertilizer and the C. pilosula stems and leaves fermented with compound microbial fertilizer can improve the soil nutrients， growth， development， yield， and quality of C. pilosula， and thus they can be applied to the artificial cultivation of C. pilosula.

ObjectiveTo study the effects of applying different microbial fertilizers on the growth and rhizosphere soil environment of Codonopsis pilosula and provide a theoretical basis for ecological cultivation of this medicinal plant. MethodsSeven groups were designed， including CK （no application of microbial fertilizer）， T1 （Trichoderma longibrachiatum fertilizer）， T2 （Bacillus subtilis fertilizer）， T3 （Trichoderma viride fertilizer）， T4 （compound microbial fertilizer）， T5 （C. pilosula stems and leaves fermented with compound microbial fertilizer）， and T6 （Scutellaria baicalensis stems and leaves fermented with T. viride fertilizer）. The physiological indicators， yield， and quality of C. pilosula and the physicochemical properties， enzyme activities， and microbial diversity in the rhizosphere soil of different fertilizer treatments were measured. ResultsGroup T1 showed slight decreases in soluble protein content （SPC） and superoxide dismutase （SOD）. Groups T2-T6 showed increases in physiological indicators such as proline （Pro）， soluble solids content （SSC）， SPC， catalase （CAT）， and peroxidase （POD） and a decrease in malondialdehyde （MDA） in C. pilosula leaves. All the fertilizer treatments increased the yield of C. pilosula and the total polysaccharide content in the roots. T1， T2， T3， T4， and T5 increased the total flavonoid content in the roots. Meanwhile， T4 increased the total saponin content in the roots. All the fertilizer treatments reduced the pH and increased the electric conductivity （EC）， soil organic matter （SOM）， and alkaline nitrogen （AN） in the soil. T2 and T5 increased the available phosphorus （AP）， and T3， T4， T5， and T6 increased the available potassium （AK） in the soil. All the fertilizer treatments increased the activities of urease， sucrase， and CAT in the soil. Except that T1 decreased the bacterial diversity in the soil， other fertilizer treatments significantly increased bacterial and fungal diversity in the soil. Different fertilizer treatments significantly affected the composition of bacterial and fungal communities in the soil. At the phylum level， the dominant bacterial phyla included Proteobacteria， Acidobacteriota， and Bacteroideta， and the dominant fungal phyla were Ascomycota， Mortierellomycota， and unclassified_fungi in the rhizosphere soil of C. pilosula after bacterial fertilizer treatment. At the genus level， unclassified Gemmatimonadaceae， Sphingomonas， and unclassified Vicinamibacteraceae were the dominant bacterial genera， while unidentified， unclassified Fungi， and unclassified Sordariomycetes were the dominant fungal genera in the rhizosphere soil. The results of redundancy analysis indicated that the main physicochemical factors affecting changes of microbial communities in the rhizosphere soil of C. pilosula were pH， EC， AK， AN， AP， and soil organic matter （SOM） in the soil. The correlation heatmap showed that Bryobacter had significantly positive correlations with EC， AK， and AN. There was a significantly negative correlation between Fusarium and SOM. In summary， applying an appropriate amount of microbial fertilizer can promote the growth and improve the rhizosphere soil environment of C. pilosula. ConclusionThe compound microbial fertilizer and the C. pilosula stems and leaves fermented with compound microbial fertilizer can improve the soil nutrients， growth， development， yield， and quality of C. pilosula， and thus they can be applied to the artificial cultivation of C. pilosula.

Objective:To investigate the predictive value of serum HOXA6 and HOXA10 levels in assessing endometrial receptivity in patients with diminished ovarian reserve (DOR).Methods:A total of 248 DOR patients treated at Xi'an People's Hospital from January 2021 to October 2023 were enrolled, with 200 healthy female volunteers as controls. Serum HOXA6 and HOXA10 concentrations were measured using enzyme-linked immunosorbent assay (ELISA). Endometrial thickness was assessed via transvaginal ultrasound on days 20-24 of the menstrual cycle, and endometrial receptivity was classified into high receptivity (≥8 mm), moderate receptivity (6-8 mm) and low receptivity (<6 mm) group. Follicle-stimulating hormone (FSH) and anti-Müllerian hormone (AMH) levels were detected by chemiluminescence immunoassay on day 3 of the menstrual cycle. Follicle count and size were recorded via transvaginal ultrasound. Normally distributed data were expressed as Mean±SD, comparison between two groups was performed by independent samples t-test, comparison among multiple groups was performed by one-way ANOVA, pairwise comparison was performed by Tukey's HSD post hoc test. Categorical data were presented as rates or percentages and analyzed by χ2 test. Logistic regression identified independent risk factors for DOR, and Pearson correlation analyzed relationships between HOXA6/HOXA10, endometrial receptivity, and ovarian reserve. Receiver operating characteristic (ROC) curves evaluated the diagnostic value of HOXA6 and HOXA10 in predicting endometrial receptivity. Results:Serum levels of both HOXA6 [(120.34±15.16) ng/L] and HOXA10 [(98.46±12.61) ng/L] in the DOR group were significantly higher than those in the control group [(80.27±10.54) ng/L and (60.36±8.80) ng/L, respectively, t=31.70, 36.20, both P<0.001]. Endometrial thickness in the DOR group [(7.4±1.3) mm] was significantly less than that in the control group [(10.0±1.1) mm]. Serum FSH levels [(12.03±2.54) U/L] were significantly higher in the DOR group compared to the control group [(8.08±2.82) U/L], while AMH levels [(1.56±0.45) μg/L] and the number of follicles [(3.52±1.63) μg/L] were significantly lower than those in the control group [(5.07±1.05) μg/L and (10.09±2.06), respectively]. The differences between the two groups for these parameters were statistically significant ( t=22.52, 15.58, 18.10, 33.58, respectively; all P<0.001). Pearson correlation analysis revealed that serum HOXA6 and HOXA10 levels were negatively correlated with endometrial thickness, AMH levels, and the number of follicles in DOR patients (HOXA6: r=-0.773, -0.647, -0.721, respectively, all P<0.001; HOXA10: r=-0.788, -0.682, -0.834, respectively, all P<0.001), and positively correlated with serum FSH levels ( r=0.667 and 0.670, respectively, both P<0.001). Multivariate logistic regression analysis indicated that serum HOXA6 ( OR=1.179, 95% CI: 1.144-1.215, P<0.001), HOXA10 ( OR=1.223, 95% CI: 1.177-1.270, P<0.001), and FSH levels ( OR=1.827, 95% CI: 1.569-2.129, P<0.001) were independent risk factors for DOR in women. Endometrial thickness ( OR=0.264, 95% CI: 0.205-0.340, P<0.001), serum AMH levels ( OR=0.126, 95% CI: 0.078-0.205, P<0.001), and the number of follicles ( OR=0.058, 95% CI: 0.024-0.143, P<0.001) were identified as protective factors against DOR. ROC curve analysis showed that the AUCs for serum HOXA6, HOXA10, and their combination in predicting poor endometrial receptivity in DOR patients were 0.770, 0.820, and 0.864, respectively. The AUC value for HOXA10 was slightly higher than that for HOXA6, and the combined prediction model had the highest AUC value. Conclusion:Serum HOXA6 and HOXA10 levels demonstrate a statistically significant association with ovarian reserve function and endometrial receptivity. These findings suggest their potential as diagnostic biomarkers for assessing endometrial receptivity in patients with diminished ovarian reserve (DOR).

Neuronal injury in glaucoma persists despite effective intraocular pressure (IOP) control, necessitating neuroprotective strategies for retinal ganglion cells (RGCs). In this study, we investigated the neuroprotective role of the γ-hydroxybutyrate analog HOCPCA in a glaucoma model, focusing on its effects on CaMKII signaling, oxidative stress, and neuroinflammatory responses. Retinal tissue from high IOP animal models was analyzed via proteomics. In vitro mouse retinal explants were subjected to elevated pressure and oxidative stress, followed by HOCPCA treatment. HOCPCA significantly mitigated the RGC loss induced by oxidative stress and elevated pressure, preserving neuronal function. It restored CaMKIIα and β levels, preserving RGC integrity, while also modulating oxidative stress and neuroinflammatory responses. These findings suggest that HOCPCA, through its interaction with CaMKII, holds promise as a neuroprotective therapy for glaucoma.
Animals ; Retinal Ganglion Cells/metabolism* ; Glaucoma/pathology* ; Oxidative Stress/drug effects* ; Neuroprotective Agents/pharmacology* ; Mice ; Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism* ; Mice, Inbred C57BL ; Disease Models, Animal ; Neuroinflammatory Diseases/drug therapy* ; Neuroprotection/drug effects* ; Male ; Intraocular Pressure/drug effects*

Hemoglobin A1c (HbA1c) has a unique structure that makes monoclonal antibody (mAb) preparation challenging. This study aims to develop a method for preparing HbA1c mAbs and establish a fluorescent immunochromatographic assay (FICA) for rapid detection of HbA1c. Three glycosylated peptides were synthesized and used to prepare complete antigens, which were identified by dot enzyme-linked immunosorbent assay (Dot-ELISA) and ultraviolet absorption spectroscopy. The complete antigens and natural HbA1c were used for cross-immunization of mice, and the optimal complete antigen was selected. The mouse with the highest serum titer was chosen for mAb preparation. The purity and specificity of the mAbs were verified, and a FICA method was developed. The optimal complete antigen, with a titer of 1:512 000, was successfully prepared and selected. Fusion with splenocytes resulted in four specific HbA1c antibodies (purity > 90%). The best antibody exhibited a binding constant (Ka) of 1.67×10¹⁰ L/mol with the antigen. Based on this antibody, a FICA method was successfully established, capable of producing results within 15 min. The method demonstrated a good linear range (3%-13% HbA1c, y=0.071 3x+0.005 6, R²=0.993 7), recovery rates of 98%-102%, precision < 10.00%, and no nonspecific reactions. Clinical testing of 210 samples showed positive agreement of 96.36%, negative agreement of 97.00%, and overall agreement of 96.68%. The receiver operating characteristic (ROC) curve analysis yielded an area under curve (AUC) of 0.980 9 [95% confidence interval (CI): 0.961 0-1.000 0], with high consistency verified in multicenter studies. We successfully developed a key technique for preparing HbA1c monoclonal antibodies and established a FICA method for rapid detection of HbA1c. It will provide an efficient and convenient detection method for the early diagnosis and long-term management of diabetes and its complications.
Antibodies, Monoclonal/biosynthesis* ; Animals ; Mice ; Glycated Hemoglobin/immunology* ; Mice, Inbred BALB C ; Humans ; Antibody Specificity ; Chromatography, Affinity/methods* ; Enzyme-Linked Immunosorbent Assay/methods* ; Female

Objective:To develop a 5-year mortality risk prediction model for patients with small cell neuroendocrine carcinoma of the cervix(SCNEC).Methods:Based on the Surveillance,Epidemiology,and End Results(SEER)database and R software version 4.3.3,variables were screened via Lasso regression,followed by multivariable logistic regression and stepwise regression to develop a 5-year mortality risk prediction model for SCNEC patients.The Akaike Information Criterion(AIC),C-index,receiver operating characteristic(ROC)curve,Hosmer-Lemeshow test,and calibration curve were employed to evaluate the model.Results:Age,M stage,surgical status,and lymph node metastasis were ultimately selected as variables to construct the 5-year mortality risk prediction model for SCNEC patients.The model demonstrated superior predictive performance compared to FIGO staging(P＜0.01).The Hosmer-Lemeshow test yielded a P-value＞0.05.The C-index values for the training and validation sets were 0.808 and 0.755,respectively,with the areas under the ROC curves of 0.826 and 0.744.The calibration curves of the model fluctuated near the diagonal line,indicating good agreement between predicted and observed outcomes.The decision curve analysis demonstrated significant clinical net benefit.Results showed that higher mortality risk was associated with advanced age,M1 status,lymph node metastasis,and lack of surgical opportunity.Conclusions:The model exhibits good discriminatory power and accuracy,providing significant benefits to patients.Enhanced management should be implemented for patients with advanced age,distant metastasis,lymph node metastasis,or ineligibility for surgery.Lymph node metastasis is an independent risk factor for 5-year mortality in patients with SCNEC.

Objective:To develop a 5-year mortality risk prediction model for patients with small cell neuroendocrine carcinoma of the cervix(SCNEC).Methods:Based on the Surveillance,Epidemiology,and End Results(SEER)database and R software version 4.3.3,variables were screened via Lasso regression,followed by multivariable logistic regression and stepwise regression to develop a 5-year mortality risk prediction model for SCNEC patients.The Akaike Information Criterion(AIC),C-index,receiver operating characteristic(ROC)curve,Hosmer-Lemeshow test,and calibration curve were employed to evaluate the model.Results:Age,M stage,surgical status,and lymph node metastasis were ultimately selected as variables to construct the 5-year mortality risk prediction model for SCNEC patients.The model demonstrated superior predictive performance compared to FIGO staging(P＜0.01).The Hosmer-Lemeshow test yielded a P-value＞0.05.The C-index values for the training and validation sets were 0.808 and 0.755,respectively,with the areas under the ROC curves of 0.826 and 0.744.The calibration curves of the model fluctuated near the diagonal line,indicating good agreement between predicted and observed outcomes.The decision curve analysis demonstrated significant clinical net benefit.Results showed that higher mortality risk was associated with advanced age,M1 status,lymph node metastasis,and lack of surgical opportunity.Conclusions:The model exhibits good discriminatory power and accuracy,providing significant benefits to patients.Enhanced management should be implemented for patients with advanced age,distant metastasis,lymph node metastasis,or ineligibility for surgery.Lymph node metastasis is an independent risk factor for 5-year mortality in patients with SCNEC.

Objective:To investigate the predictive value of serum HOXA6 and HOXA10 levels in assessing endometrial receptivity in patients with diminished ovarian reserve (DOR).Methods:A total of 248 DOR patients treated at Xi'an People's Hospital from January 2021 to October 2023 were enrolled, with 200 healthy female volunteers as controls. Serum HOXA6 and HOXA10 concentrations were measured using enzyme-linked immunosorbent assay (ELISA). Endometrial thickness was assessed via transvaginal ultrasound on days 20-24 of the menstrual cycle, and endometrial receptivity was classified into high receptivity (≥8 mm), moderate receptivity (6-8 mm) and low receptivity (<6 mm) group. Follicle-stimulating hormone (FSH) and anti-Müllerian hormone (AMH) levels were detected by chemiluminescence immunoassay on day 3 of the menstrual cycle. Follicle count and size were recorded via transvaginal ultrasound. Normally distributed data were expressed as Mean±SD, comparison between two groups was performed by independent samples t-test, comparison among multiple groups was performed by one-way ANOVA, pairwise comparison was performed by Tukey's HSD post hoc test. Categorical data were presented as rates or percentages and analyzed by χ2 test. Logistic regression identified independent risk factors for DOR, and Pearson correlation analyzed relationships between HOXA6/HOXA10, endometrial receptivity, and ovarian reserve. Receiver operating characteristic (ROC) curves evaluated the diagnostic value of HOXA6 and HOXA10 in predicting endometrial receptivity. Results:Serum levels of both HOXA6 [(120.34±15.16) ng/L] and HOXA10 [(98.46±12.61) ng/L] in the DOR group were significantly higher than those in the control group [(80.27±10.54) ng/L and (60.36±8.80) ng/L, respectively, t=31.70, 36.20, both P<0.001]. Endometrial thickness in the DOR group [(7.4±1.3) mm] was significantly less than that in the control group [(10.0±1.1) mm]. Serum FSH levels [(12.03±2.54) U/L] were significantly higher in the DOR group compared to the control group [(8.08±2.82) U/L], while AMH levels [(1.56±0.45) μg/L] and the number of follicles [(3.52±1.63) μg/L] were significantly lower than those in the control group [(5.07±1.05) μg/L and (10.09±2.06), respectively]. The differences between the two groups for these parameters were statistically significant ( t=22.52, 15.58, 18.10, 33.58, respectively; all P<0.001). Pearson correlation analysis revealed that serum HOXA6 and HOXA10 levels were negatively correlated with endometrial thickness, AMH levels, and the number of follicles in DOR patients (HOXA6: r=-0.773, -0.647, -0.721, respectively, all P<0.001; HOXA10: r=-0.788, -0.682, -0.834, respectively, all P<0.001), and positively correlated with serum FSH levels ( r=0.667 and 0.670, respectively, both P<0.001). Multivariate logistic regression analysis indicated that serum HOXA6 ( OR=1.179, 95% CI: 1.144-1.215, P<0.001), HOXA10 ( OR=1.223, 95% CI: 1.177-1.270, P<0.001), and FSH levels ( OR=1.827, 95% CI: 1.569-2.129, P<0.001) were independent risk factors for DOR in women. Endometrial thickness ( OR=0.264, 95% CI: 0.205-0.340, P<0.001), serum AMH levels ( OR=0.126, 95% CI: 0.078-0.205, P<0.001), and the number of follicles ( OR=0.058, 95% CI: 0.024-0.143, P<0.001) were identified as protective factors against DOR. ROC curve analysis showed that the AUCs for serum HOXA6, HOXA10, and their combination in predicting poor endometrial receptivity in DOR patients were 0.770, 0.820, and 0.864, respectively. The AUC value for HOXA10 was slightly higher than that for HOXA6, and the combined prediction model had the highest AUC value. Conclusion:Serum HOXA6 and HOXA10 levels demonstrate a statistically significant association with ovarian reserve function and endometrial receptivity. These findings suggest their potential as diagnostic biomarkers for assessing endometrial receptivity in patients with diminished ovarian reserve (DOR).

BACKGROUND:Hypoxic exercise can promote the degradation of body fat,and changes in the external environment can affect the circadian rhythm of animals,but the mechanisms by which changes in circadian rhythm regulate adipose tissue browning and fat degradation are unclear. OBJECTIVE:To elucidate the mechanism of clock gene regulation on epididymal adipose tissue Browning in obese rats undergoing hypoxia exercise. METHODS:Forty obese rats were randomly selected and divided into four groups(n=10 per group):normoxic sedentary group,hypoxic sedentary group,normoxic exercise group,and hypoxic exercise group for 4 weeks of intervention.The rats in the sedentary groups were not intervened,while those in the hypoxic groups lived in a hypoxic chamber with an oxygen concentration of 13.6%for the whole day.In the exercise groups,adaptive training was performed in the 1st week,and the speed and length of training remained unchanged for the last 3 weeks.The body mass,body length and perirenal fat mass of obese rats were measured.Serum levels of triacylglycerol,total cholesterol,low-density lipoprotein cholesterol,and high-density lipoprotein cholesterol in obese rats were detected by a biochemical assay kit.Liver fat content was observed by oil red O staining.Hematoxylin-eosin staining was used to evaluate the browning of epididymal adipose tissue of rats in different groups.RNA sequencing combined with bioinformatics analysis was used to analyze transcriptome changes in adipose tissue.The mRNA expressions of PGC-1α,Beclin 1,KLF 2 and Perilipin 1 in epididymal adipose tissue were detected by RT-PCR. RESULTS AND CONCLUSION:Hypoxic exercise intervention significantly decreased body mass,body fat percentage,Lee's index,serum triacylglycerol,total cholesterol,and low-density lipoprotein cholesterol levels(P＜0.01),and significantly increased high-density lipoprotein cholesterol level(P＜0.01).Oil red O staining and hematoxylin-eosin staining results showed that hypoxic exercise was more effective in promoting fat mobilization in liver tissue and promoting the browning of parepididymal adipose tissue compared with normoxic sedentary group,hypoxic sedentary group,and normoxic exercise group.RNA-seq results showed that hypoxic exercise significantly upregulated the expression of clock genes Dbp,Nr1d1,Sik1 and adipose tissue browning gene Ppargc1a(PGC-1α)and downregulated the expression of Arntl(Bmal1),accompanied by the enhanced expression of genes related to substance metabolism.qRT-PCR indicated that hypoxic exercise significantly increased the mRNA expression levels of PGC-1α and Perilipin1(P＜0.01).Therefore,these findings indicate that clock genes play an important role in promoting adipose tissue browning during hypoxic exercise.

Objective To analyze the effect of luteinizing hormone (LH) change amplitude in follicular phase on ovarian response and pregnancy outcome after fresh embryo transfer cycle in infertility patients with normal ovarian function during controlled ovarian hyperstimulation (COH) with antagonist protocol. Methods A total of 277 infertility patients who underwent in vitro fertilization and embryo transfer (IVF-ET) in the Center for Reproductive Medicine of this hospital from January 2019 to December 2021 were se-lected.Patients were divided into the high LH group (n=91),medium LH group (n=93) and low LH group (n=93) according to the tertiles of changes between LH level on the fourth day of COH and basal LH level before COH administration.The basic clinical features,use of exogenous gonadotropin (Gn) during COH, hormone levels at different periods,indexes of ovarian response,laboratory outcomes and pregnancy outcomes during fresh embryo transfer cycle were compared among all groups.Results There was no significant differ-ence in age,infertility duration,infertility types,basal follicle stimulating hormone (FSH),estradiol and tes-tosterone levels among the three groups (P>0.05).The duration of use of Gn in the high LH group was the shortest,the total amount and initiation dose of Gn was the smallest,and the estradiol level,ovarian sensitivity index (OSI),numbers of oocytes,metaphase Ⅱ(MII) oocytes,diprokaryotic fertilization,embryo formation and high-quality embryos were the highest after exogenous Gn stimulation,and the differences were statisti-cally significant compared with the other two groups (P<0.016).There was no significant difference in the clinical pregnancy rate,biochemical pregnancy rate and early miscarriage rate during fresh embryo transfer cy-cle among the three groups (P>0.05).Conclusion The infertility patients with normal ovarian function who have a large decrease in LH level during COH with antagonist protocol can obtain better embryo quantity and quality,but there was no effect on the pregnancy outcome during fresh embryo transfer cycle.