OBJECTIVE To optimize the extraction process of Shenye buyi formula. METHODS Based on the quality by design （QbD） concept， the contents of rutin， nuciferine， sinapine thiocyanate， liquiritin，and glycyrrhizic acid ammonium salt， as well as the extract yield， were selected as the critical quality attributes （CQAs） for the extraction process of Shenye buyi formula. The fishbone diagram combined with the failure model and effect analysis method was adopted to determine the water addition， extraction time and extraction times as critical process parameters （CPPs）. High-performance liquid chromatography was employed to detect the contents of rutin， nuciferine， sinapine thiocyanate， liquiritin， and glycyrrhizic acid ammonium salt. The analytic hierarchy process-entropy weight method was used to comprehensively assign weights to the content of the five components including rutin and the extract yield. The Box-Behnken response surface method was employed to fit the mathematical model between CQAs and CPPs， screen the operational space of the extraction process， and validate it. RESULTS The optimal operational space of the extraction process was 9.5 to 12 times the amount of water added， the extraction time was 1 to 1.7 hours， and the extraction times were 3. Upon verification，the comprehensive scores of the optimal extraction process within the operational space were 89.79 and 84.93， respectively. CONCLUSIONS The extraction process of the Shenye buyi formula can be applied to the development and production of its granule preparations.

Objective To elucidate the role of let-7 in the pathogenesis of oral squamous cell carcinoma(OSCC)and its underlying molecular mechanism.Methods The expression of let-7 in human oral keratinocyte(HOK),OSCC tissues,and OSCC cells(CAL-27,SCC-25,and SCC-9)was detected by quantitative real-time PCR.Colony formation,scratch,Transwell,and flow cytometry assays were used to examine the effects of let-7 on the proliferation,migration,invasion,and apoptosis of SCC-25 cells.Western blotting was used to detect the expression of Wnt and β-catenin proteins.The correlation between let-7 expression and the clinicopathological cha-racteristics and prognosis of OSCC patients was analyzed.Results let-7 was downregulated in OSCC tissues and cells.let-7 over-expression effectively inhibited the proliferation,migration,and invasion of SCC-25 cells while promoting apoptosis by suppressing acti-vation of the Wnt/β-catenin pathway.let-7 expression was closely associated with tumor stage,lymph node metastasis,and patient survival status.Patients with low let-7 expression exhibited lower overall survival rate.Conclusion let-7 targets and regulates the Wnt/β-catenin pathway,thereby inhibiting proliferation,migration,and invasion of OSCC cells while promoting apoptosis.

Objective:To establish a modified one-stage clotting assay (mOSA) based on the STA-R Evolution coagulation analyzer for quantifying emicizumab (EMI) concentration and to preliminarily evaluate its analytical performance; meanwhile to explore the clinical utility of the standard one-stage clotting assay (sOSA) in indirectly predicting EMI levels through surrogate factor Ⅷ (FⅧ) activity.Methods:A total of 30 pediatric patients with hemophilia A (HA) treated with EMI in the Hemophilia Treatment Center of Shenzhen Children′s Hospital from January 2023 to March 2025 were enrolled, and 48 post-treatment plasma samples were collected. EMI standards (2.5~100 μg/ml) were prepared using FⅧ-deficient plasma to establish the mOSA detection system. The linearity, accuracy, and precision of the method were evaluated. Surrogate FⅧ activity was measured by sOSA to estimate EMI concentrations, and its correlation with mOSA-derived EMI concentrations was analyzed using Spearman correlation analysis. The equivalent FⅧ activity in patient plasma samples was measured using a human chromogenic substrate assay-based FⅧ activity detection reagent, and Spearman correlation analysis was employed to evaluate its correlations with both the EMI concentrations measured by the mOSA method and estimated by the sOSA method respectively.Results:The established mOSA method for EMI detection showed excellent linearity in the range of 2.5?100 μg/ml ( Y=1.047 X?1.033, R 2=0.995, P<0.001). Average spike recovery rates at 25, 50, and 75 μg/ml were 101.55%(25.39/25.00), 105.31%(52.66/50.00), and 98.20%(73.65/75.00), respectively. Coefficients of variations of within-and inter-batch were 3.47%?4.80% and 6.30%?8.96%, respectively. A prediction model for EMI concentration was established as follows: estimated EMI concentration (μg/ml)=0.095×[alternative FⅧ activity (%) measured by sOSA]+2.652 ( R2=0.999, P<0.001). Validation demonstrated a strong correlation between the EMI concentration measured by the mOSA method and the EMI concentration estimated by the sOSA method ( r=0.989, P<0.001), with good consistency ( Y=1.014 X+0.684, R2=0.972, P<0.001). Both the EMI concentration measured by the mOSA method and the EMI concentration estimated by the sOSA method showed extremely strong correlations with the equivalent FⅧ activity ( r=0.986 and 0.987, respectively; P<0.001 for both). Conclusions:The mOSA system established on the STA-R Evolution analyzer demonstrates robust linearity, accuracy, and reproducibility, fulfilling clinical requirements for therapeutic drug monitoring of EMI. The sOSA method provides reliable indirect estimation of EMI concentrations through surrogate FⅧ activity, offering critical support for emergency decision-making.

Objective To explore the effect of Baofei Yiqi pill on immune function of patients with lung cancer.Methods A total of 100 patients with lung cancer diagnosed and treated in Leping Hospital of Traditional Chinese Medicine from September 2020 to September 2023 were selected and divided into experimental group and control group according to random number table method,with 50 cases in each group.The control group was treated with docetaxel combined with cisplatin,and the experimental group was treated with Baofei Yiqi pill on the basis of control group.Both groups were treated with 3 cycles of chemotherapy.The clinical efficacy,quality of life,changes in weight,immune function and adverse reactions were compared between two groups.Results The total response rate of experimental group was significantly higher than that of control group(x2=14.620,P＜0.001).After treatment,the Karnofsky performance status score and weight of experimental group were significantly higher than those of control group(P＜0.05),and the levels of CD3+,CD4+and CD4+/CD8+in experimental group were significantly higher than those in control group,and the levels of CD8+was significantly lower than than in control group(P＜0.05).The incidence of gastrointestinal reactions in experimental group was significantly lower than that in control group(P＜0.05).There was no significant difference in the incidence of abnormal renal function and liver function between two groups(P＞0.05).Conclusion Baofei Yiqi pill can effectively improve the immune function of patients with lung cancer,improve the quality of life of patients,reduce the adverse reactions of chemotherapy,and alleviate the disease of patients to a certain extent,which is worthy of promotion and application.

Objective:To investigate the clinical effect of near-infrared autofluorescence (NIRAF) imaging combined with nano-carbon in the identification of parathyroid gland.Methods:A total of 80 patients with thyroid cancer requiring bilateral thyroidectomy plus single/bilateral central area lymphatic dissection were prospectively enrolled in the Department of Thyroid and Breast Surgery of the Shenzhen Third People′s Hospital from January 2020 to June 2023. Two groups were divided in order of time of admission. In the combined group (40 cases), the parathyroid glands were identified by NIRAF+ nano-carbon negative imaging. The nano-carbon group (40 cases) was negatively developed with nano-carbon alone. The detection rate and accuracy of parathyroid gland and the changes of parathyroid hormone (1 d and 6 months after operation) were compared between the two groups.Results:There was no significant difference in the detection rate of parathyroid glands between the two groups ( P=0.65), and the accuracy rate in the combined group was higher than that in the nano-carbon group, the difference was statistically significant (χ 2=8.79, P=0.003). The level of parathyroid hormone in the combined group was higher than that in the nano-carbon group 1 day after surgery, and the difference was statistically significant ( Z=-1.19, P=0.03). There was no significant difference in parathyroid hormone levels between the two groups 6 months after operation ( P=0.18). Conclusions:NIRAF imaging combined with nano-carbon negative imaging can improve the accuracy of intraoperative parathyroid identification, and may reduce the incidence of temporary postoperative hypoparathyroidism.

Objective To elucidate the role of let-7 in the pathogenesis of oral squamous cell carcinoma(OSCC)and its underlying molecular mechanism.Methods The expression of let-7 in human oral keratinocyte(HOK),OSCC tissues,and OSCC cells(CAL-27,SCC-25,and SCC-9)was detected by quantitative real-time PCR.Colony formation,scratch,Transwell,and flow cytometry assays were used to examine the effects of let-7 on the proliferation,migration,invasion,and apoptosis of SCC-25 cells.Western blotting was used to detect the expression of Wnt and β-catenin proteins.The correlation between let-7 expression and the clinicopathological cha-racteristics and prognosis of OSCC patients was analyzed.Results let-7 was downregulated in OSCC tissues and cells.let-7 over-expression effectively inhibited the proliferation,migration,and invasion of SCC-25 cells while promoting apoptosis by suppressing acti-vation of the Wnt/β-catenin pathway.let-7 expression was closely associated with tumor stage,lymph node metastasis,and patient survival status.Patients with low let-7 expression exhibited lower overall survival rate.Conclusion let-7 targets and regulates the Wnt/β-catenin pathway,thereby inhibiting proliferation,migration,and invasion of OSCC cells while promoting apoptosis.

Objective To explore the effect of Baofei Yiqi pill on immune function of patients with lung cancer.Methods A total of 100 patients with lung cancer diagnosed and treated in Leping Hospital of Traditional Chinese Medicine from September 2020 to September 2023 were selected and divided into experimental group and control group according to random number table method,with 50 cases in each group.The control group was treated with docetaxel combined with cisplatin,and the experimental group was treated with Baofei Yiqi pill on the basis of control group.Both groups were treated with 3 cycles of chemotherapy.The clinical efficacy,quality of life,changes in weight,immune function and adverse reactions were compared between two groups.Results The total response rate of experimental group was significantly higher than that of control group(x2=14.620,P＜0.001).After treatment,the Karnofsky performance status score and weight of experimental group were significantly higher than those of control group(P＜0.05),and the levels of CD3+,CD4+and CD4+/CD8+in experimental group were significantly higher than those in control group,and the levels of CD8+was significantly lower than than in control group(P＜0.05).The incidence of gastrointestinal reactions in experimental group was significantly lower than that in control group(P＜0.05).There was no significant difference in the incidence of abnormal renal function and liver function between two groups(P＞0.05).Conclusion Baofei Yiqi pill can effectively improve the immune function of patients with lung cancer,improve the quality of life of patients,reduce the adverse reactions of chemotherapy,and alleviate the disease of patients to a certain extent,which is worthy of promotion and application.

Objective:To investigate the clinical effect of near-infrared autofluorescence (NIRAF) imaging combined with nano-carbon in the identification of parathyroid gland.Methods:A total of 80 patients with thyroid cancer requiring bilateral thyroidectomy plus single/bilateral central area lymphatic dissection were prospectively enrolled in the Department of Thyroid and Breast Surgery of the Shenzhen Third People′s Hospital from January 2020 to June 2023. Two groups were divided in order of time of admission. In the combined group (40 cases), the parathyroid glands were identified by NIRAF+ nano-carbon negative imaging. The nano-carbon group (40 cases) was negatively developed with nano-carbon alone. The detection rate and accuracy of parathyroid gland and the changes of parathyroid hormone (1 d and 6 months after operation) were compared between the two groups.Results:There was no significant difference in the detection rate of parathyroid glands between the two groups ( P=0.65), and the accuracy rate in the combined group was higher than that in the nano-carbon group, the difference was statistically significant (χ 2=8.79, P=0.003). The level of parathyroid hormone in the combined group was higher than that in the nano-carbon group 1 day after surgery, and the difference was statistically significant ( Z=-1.19, P=0.03). There was no significant difference in parathyroid hormone levels between the two groups 6 months after operation ( P=0.18). Conclusions:NIRAF imaging combined with nano-carbon negative imaging can improve the accuracy of intraoperative parathyroid identification, and may reduce the incidence of temporary postoperative hypoparathyroidism.

Objective:To establish a modified one-stage clotting assay (mOSA) based on the STA-R Evolution coagulation analyzer for quantifying emicizumab (EMI) concentration and to preliminarily evaluate its analytical performance; meanwhile to explore the clinical utility of the standard one-stage clotting assay (sOSA) in indirectly predicting EMI levels through surrogate factor Ⅷ (FⅧ) activity.Methods:A total of 30 pediatric patients with hemophilia A (HA) treated with EMI in the Hemophilia Treatment Center of Shenzhen Children′s Hospital from January 2023 to March 2025 were enrolled, and 48 post-treatment plasma samples were collected. EMI standards (2.5~100 μg/ml) were prepared using FⅧ-deficient plasma to establish the mOSA detection system. The linearity, accuracy, and precision of the method were evaluated. Surrogate FⅧ activity was measured by sOSA to estimate EMI concentrations, and its correlation with mOSA-derived EMI concentrations was analyzed using Spearman correlation analysis. The equivalent FⅧ activity in patient plasma samples was measured using a human chromogenic substrate assay-based FⅧ activity detection reagent, and Spearman correlation analysis was employed to evaluate its correlations with both the EMI concentrations measured by the mOSA method and estimated by the sOSA method respectively.Results:The established mOSA method for EMI detection showed excellent linearity in the range of 2.5?100 μg/ml ( Y=1.047 X?1.033, R 2=0.995, P<0.001). Average spike recovery rates at 25, 50, and 75 μg/ml were 101.55%(25.39/25.00), 105.31%(52.66/50.00), and 98.20%(73.65/75.00), respectively. Coefficients of variations of within-and inter-batch were 3.47%?4.80% and 6.30%?8.96%, respectively. A prediction model for EMI concentration was established as follows: estimated EMI concentration (μg/ml)=0.095×[alternative FⅧ activity (%) measured by sOSA]+2.652 ( R2=0.999, P<0.001). Validation demonstrated a strong correlation between the EMI concentration measured by the mOSA method and the EMI concentration estimated by the sOSA method ( r=0.989, P<0.001), with good consistency ( Y=1.014 X+0.684, R2=0.972, P<0.001). Both the EMI concentration measured by the mOSA method and the EMI concentration estimated by the sOSA method showed extremely strong correlations with the equivalent FⅧ activity ( r=0.986 and 0.987, respectively; P<0.001 for both). Conclusions:The mOSA system established on the STA-R Evolution analyzer demonstrates robust linearity, accuracy, and reproducibility, fulfilling clinical requirements for therapeutic drug monitoring of EMI. The sOSA method provides reliable indirect estimation of EMI concentrations through surrogate FⅧ activity, offering critical support for emergency decision-making.