ObjectiveBased on multi-index and multi-method detection， the quality of Rukuaixiao preparations was systematically evaluated from the perspective of characteristic components， and the existing problems were analyzed. MethodsLiquid chromatography-mass spectrometry（LC-MS） for the determination of 16 characteristic components was established to evaluate the quality of 129 batches of Rukuaixiao preparations. High performance liquid chromatography（HPLC） was established to determine the contents of salvianolic acids and tanshinones， investigate the rationality of quality control index of Salviae Miltiorrhizae Radix et Rhizoma in the standard for Rukuaixiao preparations in the 2020 edition of Pharmacopoeia of the People's Republic of China（hereinafter referred to as Chinese Pharmacopoeia） （volume Ⅰ）， and trace the causes of significant difference among different batches. The processing and different extraction methods of Vaccariae Semen were tested， analyzing the impact of formulation changes across different editions of Chinese Pharmacopoeia. The LC-MS was established for determining the changes in the ratio of toosendanin and isotoosendanin after water extraction of Toosendan Fructus. The contents of active components in different parts of Gleditsiae Spina were determined to identify the reason of the low contents of characteristic components in some enterprises. ResultsBased on the comprehensive analysis of the samples from different dosage forms， the contents of characteristic components of Vaccariae Semen and Gleditsiae Spina in tablets from manufacturer B and granules from manufacturer D were significantly higher than those in tablets from manufacturer A， and tablets and capsules from manufacturer C. The contents of tanshinones in some batches of products from manufacturer A were abnormally high， potentially linked to the use of 70% ethanol reflux during extraction of Salviae Miltiorrhizae Radix et Rhizoma. All samples from manufacturer C exhibited abnormally high proportions of salvianic acid A and 3，4-dihydroxybenzaldehyde（salvianolic acid degradation products） to the total seven phenolic acids， indicating a risk of blindly pursuing tanshinol content and neglecting more effective components. The extraction rate of the characteristic components from Vaccariae Semen by decocting with water was obviously higher than that by reflux extraction with 70% ethanol. However， using the stir-fried Vaccariae Semen could reduce the loss of ingredients. The content ratio of toosendanin and isotoosendanin decreased from the crude herb to the prepared medicine when Toosendan Fructus was prepared by water decoction. The reason for the low component content of Gleditsiae Spina may be attributed to manufacturers using excessive non-medicinal parts in their formulations. ConclusionIt is suggested that enterprises should ensure the quality of raw material inputs， especially those without quality-control items in the standard， reduce the use of non-medicinal parts， and prohibit arbitrary alterations to manufacturing methods or processes. It is also recommended that Vaccariae Semen in Rukuaixiao capsules and granules should be changed to the stir-fried processed products. Isotoosendanin should be taken into consideration in the drug supervision when Toosendan Fructus is prepared into medicine by water decoction. Salvianolic acid B should be set as a quality control index for Salviae Miltiorrhizae Radix et Rhizoma when revising the pharmacopoeia standard of Rukuaixiao preparations.

ObjectiveBased on multi-index and multi-method detection， the quality of Rukuaixiao preparations was systematically evaluated from the perspective of characteristic components， and the existing problems were analyzed. MethodsLiquid chromatography-mass spectrometry（LC-MS） for the determination of 16 characteristic components was established to evaluate the quality of 129 batches of Rukuaixiao preparations. High performance liquid chromatography（HPLC） was established to determine the contents of salvianolic acids and tanshinones， investigate the rationality of quality control index of Salviae Miltiorrhizae Radix et Rhizoma in the standard for Rukuaixiao preparations in the 2020 edition of Pharmacopoeia of the People's Republic of China（hereinafter referred to as Chinese Pharmacopoeia） （volume Ⅰ）， and trace the causes of significant difference among different batches. The processing and different extraction methods of Vaccariae Semen were tested， analyzing the impact of formulation changes across different editions of Chinese Pharmacopoeia. The LC-MS was established for determining the changes in the ratio of toosendanin and isotoosendanin after water extraction of Toosendan Fructus. The contents of active components in different parts of Gleditsiae Spina were determined to identify the reason of the low contents of characteristic components in some enterprises. ResultsBased on the comprehensive analysis of the samples from different dosage forms， the contents of characteristic components of Vaccariae Semen and Gleditsiae Spina in tablets from manufacturer B and granules from manufacturer D were significantly higher than those in tablets from manufacturer A， and tablets and capsules from manufacturer C. The contents of tanshinones in some batches of products from manufacturer A were abnormally high， potentially linked to the use of 70% ethanol reflux during extraction of Salviae Miltiorrhizae Radix et Rhizoma. All samples from manufacturer C exhibited abnormally high proportions of salvianic acid A and 3，4-dihydroxybenzaldehyde（salvianolic acid degradation products） to the total seven phenolic acids， indicating a risk of blindly pursuing tanshinol content and neglecting more effective components. The extraction rate of the characteristic components from Vaccariae Semen by decocting with water was obviously higher than that by reflux extraction with 70% ethanol. However， using the stir-fried Vaccariae Semen could reduce the loss of ingredients. The content ratio of toosendanin and isotoosendanin decreased from the crude herb to the prepared medicine when Toosendan Fructus was prepared by water decoction. The reason for the low component content of Gleditsiae Spina may be attributed to manufacturers using excessive non-medicinal parts in their formulations. ConclusionIt is suggested that enterprises should ensure the quality of raw material inputs， especially those without quality-control items in the standard， reduce the use of non-medicinal parts， and prohibit arbitrary alterations to manufacturing methods or processes. It is also recommended that Vaccariae Semen in Rukuaixiao capsules and granules should be changed to the stir-fried processed products. Isotoosendanin should be taken into consideration in the drug supervision when Toosendan Fructus is prepared into medicine by water decoction. Salvianolic acid B should be set as a quality control index for Salviae Miltiorrhizae Radix et Rhizoma when revising the pharmacopoeia standard of Rukuaixiao preparations.

Objective To explore the effect of paeoniflorin on the inflammatory response in rats with periapical periodontitis by regulating the TLR4/MyD88/NF-κB pathway.Methods Periapical periodontitis model was established in 72 rats,which were then randomly divided into 6 groups:model group,model+low-dose paeoniflorin(model+paeoniflorin-L,5 mg/kg)group,model+medium-dose paeoniflorin(model+paeoniflorin-M,10 mg/kg)group,model+high-dose paeoniflorin(model+paeoniflorin-H,20 mg/kg)group,and model+paeoniflorin-H+TLR4 activator(LPS)group(model+paeoniflorin-H+LPS,1 μg/kg LPS),with 12 rats in each group.In addition,12 normal rats were included as the control check(CK)group.The levels of CRP,IL-6 and IL-2 were detected by ELISA;the damage of periapical tissues was observed by HE staining;the number of tartrate-resistant acid phosphatase(TRAP)-positive osteoclasts was detected by TRAP staining;the level of cathepsin K and the proteins related to the TLR4/MyD88/NF-κB pathway were detected by Western blotting.Results There were very few inflammatory cells in the CK group,while there was a large accumulation of inflammatory cells in the model group.The expression levels of CRP,IL-6,IL-2,tissue protease K,osteoclast positive cells,TLR4,MyD88,p-NF-κB p65/NF-κB p65 proteins in the model group were higher than those in the CK group(P＜0.05).Compared with the model group,the above-mentioned indicators in the model+paeoniflorin-L group,model+paeoniflorin-M group,and model+paeoniflorin-H group decreased successively(P＜0.05).Compared with the model+paeoniflorin-H group,the above-mentioned indicators in the model+paeoniflorin-H+LPS group increased(P＜0.05).Conclusion Paeoniflorin may alleviate the inflammatory response in rats with periapical periodontitis by inhibiting the TLR4/MyD88/NF-κB signaling pathway.

Objective:To analyze the quality stability of Ruduxiao preparations from both process stability and for-mulation stability.Methods:HPLC fingerprints of different formulations of Rutuxiaoqi were established.According to the fingerprint screening results,the abnormal content of tansanone in some batches of manufacturer A and the pollution components of manufacturer B for non-medicinal purposes were tracked.Accelerated stability tests were performed on cross-manufacturer samples,and evaluated via multifaceted indicators including description,moisture content and fingerprint profiling.Results:A total of 35 common peaks were identified in the fingerprint,and the reference fingerprints of tablet and granule were close to each other,but the difference was obvious from that of cap-sule.The main difference lies in that some peaks attributed to Salvia Miltiorrhiza in the reference fingerprint of the capsules produced by manufacturer C were partially missing and their contents were relatively low.There were devi-ations in the extraction process of Salvia Miltiorrhiza from tablets of manufacturer A,and the contaminated compo-nent of manufacturer B was confirmed to be scutellaria extract powder by comparison of reference substances and UPLC Q-TOF MS/MS,which may come from collinear pollution.In the accelerated stability test,the granules packaged in the composite film of manufacturer D showed stable performance.The tablets packaged with aluminum-plastic blister were significantly affected by water vapor,and appeared splits,contents overflow,softening,etc.The weight of the tablets increases significantly,and the common peak 1 of fingerprint was significantly degraded,while the tablets and capsules packaged with HDPE plastic bottles were degraded obviously.Conclusion:The established fingerprint method can effectively characterize the overall situation of Ruduxiao preparations,not only can detect the abnormal process changes and non-medicinal component residues,but also can reveal the sensitive conditions that affect the stability of the samples,providing a reference for process verification and upgrading of packaging materials for manufacturers.The peak of common peak 1 can be used as an indicator of the stability of Ruduxiao preparations.

Objective:To investigate the predictive value of nutritional risk related indicators in the prognosis of elderly esophageal squamous cell carcinoma (ESCC) patients undergoing radical radiotherapy.Methods:A retrospective case series study was conducted. The clinical data of ESCC patients aged ≥ 70 years who received radical radiotherapy in Shanxi Province Cancer Hospital from January 2015 to December 2019 were retrospectively analyzed. According to the radiotherapy planning system, the maximum transverse diameter of gross tumor volume of primary tumors (GTVt) and GTVt volume in the esophagus were calculated. The nutritional risk related indicators of ESCC patients before and after radiotherapy were calculated and recorded, including body mass index (BMI), geriatric nutritional risk index (GNRI), and neutrophil-to-lymphocyte ratio (NLR); Kaplan-Meier method was used for survival analysis, and log- rank test was performed; Cox proportional risk model was used to analyze the factors influencing the prognosis of patients. The overall survival (OS), progression-free survival (PFS), short-term efficacy, and adverse reactions of patients with different clinicopathological characteristics were compared.Results:The last follow-up time was 30 December, 2023. The 1-year, 3-year, and 5-year OS rates of 161 elderly patients with ESCC were 79.5%, 33.9%, and 16.1% respectively, with a median OS time of 25.8 months (95% CI: 20.11-31.49 months); the 1-year, 3-year, and 5-year PFS rates were 65.8%, 28.9%, and 14.8%, respectively, with a median PFS time of 20.0 months (95% CI: 16.31-23.69 months). There were statistically significant differences in the median OS time of patients with different age, lesion contrast length, maximum transverse diameter of GTVt, GTVt volume, short-term efficacy, BMI before radiotherapy, BMI after radiotherapy, GNRI after radiotherapy, NLR before radiotherapy, and NLR after radiotherapy (all P < 0.05); there were statistically significant differences in the median PFS time of ESCC patients with different age, maximum transverse diameter of GTVt, GTVt volume, short-term efficacy, BMI before radiotherapy, BMI after radiotherapy, GNRI after radiotherapy, and NLR before radiotherapy (all P <0.05). The results of multivariate Cox regression analysis showed that the maximum transverse diameter of GTVt was an independent influencing factor of the patients' OS ( P < 0.05); GTVt volume, short-term efficacy, GNRI after radiotherapy, and NLR before radiotherapy were independent influencing factors for OS and PFS of the patients (all P < 0.05). Among 161 elderly patients with ESCC, 45 achieved complete remission (CR), 111 achieved partial remission (PR), and 5 achieved stable disease (SD). There were statistically significant differences in the proportions of patients with different maximum transverse diameter of GTVt, GTVt volume, BMI after radiotherapy, GNRI after radiotherapy, and NLR before radiotherapy reaching CR and PR+SD (all P < 0.05). Among the 161 patients, grade 0, 1, 2, and 3 radiation-induced esophagitis (RE) occurred in 59, 54, 42, and 6 cases, respectively during treatment and within 3 months after treatment; among them, ≥ grade 2 RE occurred in 48 cases (29.8%); grade 0, 1, 2, and 3 radiation pneumonitis (RP) occurred in 95, 38, 25, and 3 cases, respectively; among them, ≥ grade 2 RP occurred in 28 cases (17.4%). Conclusions:Nutritional risk related indexes GNRI and NLR may be predictive indicators for the prognosis of elderly ESCC patients undergoing radical radiotherapy.

Objective:To analyze the quality stability of Ruduxiao preparations from both process stability and for-mulation stability.Methods:HPLC fingerprints of different formulations of Rutuxiaoqi were established.According to the fingerprint screening results,the abnormal content of tansanone in some batches of manufacturer A and the pollution components of manufacturer B for non-medicinal purposes were tracked.Accelerated stability tests were performed on cross-manufacturer samples,and evaluated via multifaceted indicators including description,moisture content and fingerprint profiling.Results:A total of 35 common peaks were identified in the fingerprint,and the reference fingerprints of tablet and granule were close to each other,but the difference was obvious from that of cap-sule.The main difference lies in that some peaks attributed to Salvia Miltiorrhiza in the reference fingerprint of the capsules produced by manufacturer C were partially missing and their contents were relatively low.There were devi-ations in the extraction process of Salvia Miltiorrhiza from tablets of manufacturer A,and the contaminated compo-nent of manufacturer B was confirmed to be scutellaria extract powder by comparison of reference substances and UPLC Q-TOF MS/MS,which may come from collinear pollution.In the accelerated stability test,the granules packaged in the composite film of manufacturer D showed stable performance.The tablets packaged with aluminum-plastic blister were significantly affected by water vapor,and appeared splits,contents overflow,softening,etc.The weight of the tablets increases significantly,and the common peak 1 of fingerprint was significantly degraded,while the tablets and capsules packaged with HDPE plastic bottles were degraded obviously.Conclusion:The established fingerprint method can effectively characterize the overall situation of Ruduxiao preparations,not only can detect the abnormal process changes and non-medicinal component residues,but also can reveal the sensitive conditions that affect the stability of the samples,providing a reference for process verification and upgrading of packaging materials for manufacturers.The peak of common peak 1 can be used as an indicator of the stability of Ruduxiao preparations.

Objective To explore the effect of paeoniflorin on the inflammatory response in rats with periapical periodontitis by regulating the TLR4/MyD88/NF-κB pathway.Methods Periapical periodontitis model was established in 72 rats,which were then randomly divided into 6 groups:model group,model+low-dose paeoniflorin(model+paeoniflorin-L,5 mg/kg)group,model+medium-dose paeoniflorin(model+paeoniflorin-M,10 mg/kg)group,model+high-dose paeoniflorin(model+paeoniflorin-H,20 mg/kg)group,and model+paeoniflorin-H+TLR4 activator(LPS)group(model+paeoniflorin-H+LPS,1 μg/kg LPS),with 12 rats in each group.In addition,12 normal rats were included as the control check(CK)group.The levels of CRP,IL-6 and IL-2 were detected by ELISA;the damage of periapical tissues was observed by HE staining;the number of tartrate-resistant acid phosphatase(TRAP)-positive osteoclasts was detected by TRAP staining;the level of cathepsin K and the proteins related to the TLR4/MyD88/NF-κB pathway were detected by Western blotting.Results There were very few inflammatory cells in the CK group,while there was a large accumulation of inflammatory cells in the model group.The expression levels of CRP,IL-6,IL-2,tissue protease K,osteoclast positive cells,TLR4,MyD88,p-NF-κB p65/NF-κB p65 proteins in the model group were higher than those in the CK group(P＜0.05).Compared with the model group,the above-mentioned indicators in the model+paeoniflorin-L group,model+paeoniflorin-M group,and model+paeoniflorin-H group decreased successively(P＜0.05).Compared with the model+paeoniflorin-H group,the above-mentioned indicators in the model+paeoniflorin-H+LPS group increased(P＜0.05).Conclusion Paeoniflorin may alleviate the inflammatory response in rats with periapical periodontitis by inhibiting the TLR4/MyD88/NF-κB signaling pathway.

Objective:To investigate the predictive value of nutritional risk related indicators in the prognosis of elderly esophageal squamous cell carcinoma (ESCC) patients undergoing radical radiotherapy.Methods:A retrospective case series study was conducted. The clinical data of ESCC patients aged ≥ 70 years who received radical radiotherapy in Shanxi Province Cancer Hospital from January 2015 to December 2019 were retrospectively analyzed. According to the radiotherapy planning system, the maximum transverse diameter of gross tumor volume of primary tumors (GTVt) and GTVt volume in the esophagus were calculated. The nutritional risk related indicators of ESCC patients before and after radiotherapy were calculated and recorded, including body mass index (BMI), geriatric nutritional risk index (GNRI), and neutrophil-to-lymphocyte ratio (NLR); Kaplan-Meier method was used for survival analysis, and log- rank test was performed; Cox proportional risk model was used to analyze the factors influencing the prognosis of patients. The overall survival (OS), progression-free survival (PFS), short-term efficacy, and adverse reactions of patients with different clinicopathological characteristics were compared.Results:The last follow-up time was 30 December, 2023. The 1-year, 3-year, and 5-year OS rates of 161 elderly patients with ESCC were 79.5%, 33.9%, and 16.1% respectively, with a median OS time of 25.8 months (95% CI: 20.11-31.49 months); the 1-year, 3-year, and 5-year PFS rates were 65.8%, 28.9%, and 14.8%, respectively, with a median PFS time of 20.0 months (95% CI: 16.31-23.69 months). There were statistically significant differences in the median OS time of patients with different age, lesion contrast length, maximum transverse diameter of GTVt, GTVt volume, short-term efficacy, BMI before radiotherapy, BMI after radiotherapy, GNRI after radiotherapy, NLR before radiotherapy, and NLR after radiotherapy (all P < 0.05); there were statistically significant differences in the median PFS time of ESCC patients with different age, maximum transverse diameter of GTVt, GTVt volume, short-term efficacy, BMI before radiotherapy, BMI after radiotherapy, GNRI after radiotherapy, and NLR before radiotherapy (all P <0.05). The results of multivariate Cox regression analysis showed that the maximum transverse diameter of GTVt was an independent influencing factor of the patients' OS ( P < 0.05); GTVt volume, short-term efficacy, GNRI after radiotherapy, and NLR before radiotherapy were independent influencing factors for OS and PFS of the patients (all P < 0.05). Among 161 elderly patients with ESCC, 45 achieved complete remission (CR), 111 achieved partial remission (PR), and 5 achieved stable disease (SD). There were statistically significant differences in the proportions of patients with different maximum transverse diameter of GTVt, GTVt volume, BMI after radiotherapy, GNRI after radiotherapy, and NLR before radiotherapy reaching CR and PR+SD (all P < 0.05). Among the 161 patients, grade 0, 1, 2, and 3 radiation-induced esophagitis (RE) occurred in 59, 54, 42, and 6 cases, respectively during treatment and within 3 months after treatment; among them, ≥ grade 2 RE occurred in 48 cases (29.8%); grade 0, 1, 2, and 3 radiation pneumonitis (RP) occurred in 95, 38, 25, and 3 cases, respectively; among them, ≥ grade 2 RP occurred in 28 cases (17.4%). Conclusions:Nutritional risk related indexes GNRI and NLR may be predictive indicators for the prognosis of elderly ESCC patients undergoing radical radiotherapy.

Objective To investigate the effects of platelet-rich plasma-derived exosomes(PRP-Exos)on the proliferation and migration of tendon stem/progenitor cell(TSPC).Methods PRP-Exos were extracted through the combination of polymer-based precipitation and ultracentrifugation.The morphology,concentration,and particle size of PRP-Exos were identified by transmission electron microscopy and nanoparticle tracking analysis.The expression levels of surface marker proteins on PRP-Exos and platelet membrane glycoproteins were deter-mined by Western blot analysis.Rat TSPC was extracted and cultured,and the expression of surface marker mol-ecules on TSPC was detected using flow cytometry and immunofluorescence staining.The proliferation of TSPC in-fluenced by PRP-Exos was evaluated using CCK-8 assay and EdU assay.The effect of PRP-Exos on the migration of TSPC was evaluated by cell scratch assay and Transwell assay.Results The extracted PRP-Exos exhibit typi-cal saucer-like structures,with a concentration of 4.9 ×1011 particles/mL,an average particle size of(132.2±56.8)nm,and surface expression of CD9,CD63 and CD41.The extracted TSPC expressed the CD44 pro-tein.PRP-Exos can be taken up by TSPC,and after co-cultured for 48 h,concentrations of 50 and 100 μg/mL of PRP-Exos significantly promoted the proliferation of TSPC(both P＜0.001),with no statistical difference be-tween the two concentrations(P=0.283).Additionally,after co-cultured for 24 h,50 μg/mL of PRP-Exos significantly promoted the migration of TSPC(P＜0.001).Conclusion Under in vitro culture conditions,PRP-Exos significantly promote the proliferation and migration of rat TSPC.

Objective:To detect the abnormal changes of myocardial blood perfusion in patients with hypertrophic cardiomyopathy(HCM) by myocardial contrast echocardiography (MCE) combined with adenosine stress test.Methods:Fifteen adult patients with HCM who were treated in Fuwai Central China Cardiovascular Hospital from May 2021 to March 2022 were prospectively selected as the HCM group, and eighteen healthy volunteers matched by gender, age and body surface area during the same period were chosen as the control group. All subjects underwent routine echocardiography, rest and adenosine stress MCE. The MCE images were analyzed by QLab software to obtain the myocardial perfusion parameters: peak signal intensity (A value), rising slope of the curve (β value) and A×β value, and the differences of above parameters between the two groups were compared.According to whether the end-diastolic wall thickness ≥12 mm, the myocardial segments in the HCM group were divided into hypertrophic segments and non-hypertrophic segments. The differences in myocardial perfusion parameters were compared among control group segments, hypertrophic segments and non-hypertrophic segments of the HCM group. The correlations of stress myocardial blood flow with maximal left ventricular wall thickness (MLVWT), left ventricular mass index (LVMI) and left atrial volume index (LAVI) in the HCM group were analyzed.Results:Compared with the control group, the A value, β value and A×β value of whole myocardium, hypertrophic segments and non-hypertrophic segments in the HCM group were significantly decreased in the rest and adenosine stress state, and the differences were statistically significant (all P<0.05). In the stress state, the A value, β value and A×β value of the hypertrophic segments were significantly lower than those in the non-hypertrophic segments in the HCM group, and the detection rate of abnormal perfusion segments in the HCM group was significantly higher than that in the rest state(all P<0.05). Compared with the control group, the myocardial blood flow reserve of whole myocardium, hypertrophic segments and non-hypertrophic segments in the HCM group were significantly decreased, and the differences were statistically significant(all P<0.05). The stress myocardial blood flow in the HCM group was negatively correlated with MLVWT, LVMI and LAVI ( r=-0.815, -0.805, -0.742; all P<0.05). Conclusions:Myocardial blood perfusion abnormalities can occur in both hypertrophic and non-hypertrophic myocardial segments in patients with HCM, and adenosine stress MCE can significantly improve the sensitivity of detecting myocardial perfusion abnormalities. The stress myocardial blood flow in patients with HCM is negatively correlated with MLVWT, LVMI and LAVI.