ObjectiveTo explore the molecular mechanism of Buzhong Yiqitang in attenuating cisplatin resistance in non-small cell lung cancer （NSCLC） by inducing ferroptosis via poly（rC）-binding protein 1 （PCBP1）. MethodsThe serum containing Buzhong Yiqitang was prepared and cisplatin-resistant human non-small cell lung cancer （NSCLC） cells （A549/DDP） were cultured and randomly grouped as follows： Blank （10% blank serum）， model （10% blank serum+20 mg·L^-1 cisplatin）， Buzhong Yiqitang （10% serum containing Buzhong Yiqitang+20 mg·L^-1 cisplatin）， Fe-1 （10% blank serum+20 mg·L^-1 cisplatin+5 μmol·L^-1 Fe-1）， and Buzhong Yiqitang+Fe-1 （10% serum containing Buzhong Yiqitang+20 mg·L^-1 cisplatin+5 μmol·L^-1 Fe-1）. Firstly， PCR Array was used to screen ferroptosis-related genes regulated by Buzhong Yiqitang， and PCBP1 was identified as the target for studying the attenuation of cisplatin resistance by Buzhong Yiqitang. Subsequently， the median inhibitory concentration （IC₅₀） of cisplatin in each group was determined by the cell counting kit-8 （CCK-8） method and the resistance index （RI） was calculated. The ultrastructure of A549/DDP cells in each group was observed by transmission electron microscopy. The protein levels of PCBP1 and glutathione peroxidase 4 （GPX4） were determined by Western blot. The lipid reactive oxygen species （ROS） content in each group was determined by the C11-BODIRY 581/591 fluorescence probe. The ferrous ion assay kit was used to measure the ferrous ion content in each group. The malondialdehyde （MDA） assay kit was used to determine the MDA content in each group. ResultsCompared with model group， the IC₅₀ of cisplatin and the RI of A549/DDP cells decreased in the Buzhong Yiqitang group （P<0.05） but increased in the Fe-1 group （P<0.05）. The IC₅₀ of cisplatin and the RI of A549/DDP cells in the Buzhong Yiqitang+Fe-1 group were lower than those in the Fe-1 group （P<0.05）. Compared with the model group， the Buzhong Yiqitang group showed obvious mitochondrial ferroptosis， while the mitochondrial damage became less obvious after Fe-1 treatment. Compared with that in the Fe-1 group， the mitochondrial ferroptosis was aggravated after the intervention with Buzhong Yiqitang. Compared with blank group， the model group showed down-regulated expression levels of PCBP1 and GPX4 （P<0.05） and increased content of lipid ROS， ferrous ions， and MDA （P<0.05） in A549/DDP cells. Compared with model group， the Buzhong Yiqitang group showed down-regulated expression levels of PCBP1 and GPX4 （P<0.05） and increased content of lipid ROS， ferrous ions， and MDA （P<0.05）， while the Fe-1 group showed up-regulated expression levels of PCBP1 and GPX4 （P<0.05） and reduced content of lipid ROS， ferrous ions， and MDA （P<0.05）. Compared with the Fe-1 group， the Buzhong Yiqitang+Fe-1 group showed down-regulated expression levels of PCBP1 and GPX4 and increased content of lipid ROS， ferrous ions， and MDA （P<0.05）. ConclusionBuzhong Yiqitang attenuated cisplatin resistance in NSCLC by regulating PCBP1 to induce ferroptosis.

Venous system diseases mainly include varicose veins and venous malformations of lower limbs and the genital system. Most of them are chronic diseases that cause serious clinical symptoms to patients and affect their health and quality of life. Sclerotherapy has become the first-line therapy for venous system diseases. However, there are problems such as incomplete fibrosis and vascular recanalization after sclerotherapy, and improper operation will cause serious adverse consequences. Therefore, exploring a safe and effective sclerotherapy strategy is essential for developing clinically successful sclerotherapy. To solve the above problems, we proposed a new sclerotherapy strategy with a dual mechanism of "vascular damage and plasmin (PLA) system inhibition." We intended to construct a novel cationic surfactant (AEO_x-TA) by reacting tranexamic acid (TA), a parent structure, with fatty alcohol polyoxyethylene ether (AEO_x) by ester bonds. AEO_x-TA could damage vascular endothelium and initiate a coagulation cascade effect to induce thrombus. Furthermore, AEO_x-TA could be degraded by esterase and release the parent drug, TA, which could inhibit the PLA system to inhibit the degradation of thrombus and extracellular matrix and promote the process of vascular fibrosis. In addition, such surfactant-based sclerosants have foam-forming properties, and they can be blended with polyvinyl alcohol (PVA) to prepare a highly stable foam formulation (AEO_x-TA/P), which can achieve a precise drug delivery and prolonged drug retention time, thereby improving drug efficacy and reducing the risk of ectopic embolism. Overall, the novel cationic surfactant AEO_x-TA provides a new avenue to resolve the bottleneck: surfactant sclerosants' efficiency is relatively low in the current sclerotherapy.

Objective To explore the effect of fluid shear stress on the expression of glucose regulatory protein 78(GRP78)and C/EBP homologous protein(CHOP)in human umbilical vein endothelial cells(HUVECs).Methods HUVECs were used as experimental cells,and a fluid dynamics simulation experimental system was designed and constructed.Different fluid shear stresses were applied to the experimental cells by controlling the flow rate of the perfusion fluid in the experimental system.According to the fluid shear stress experienced by the experimental cells in the experimental system,they were divided into low shear stress group(group A;0.4Pa),medium shear stress group(group B;0.8 Pa)and high shear stress group(group C;1.2 Pa).Each group of HUVECs consisted of 3 cell slides,and each slide was repeatedly circulated through the perfusion solution of the experimental system for 12 h.Western blotting was used to detect the levels of GRP78 and CHOP proteins,and real-time quantitative reverse transcription polymerase chain reaction was used to determine the levels of GRP78 and CHOP and their messenger RNA(mRNA)in each group.GraphPad Prism 8.0 software was used to statistically analyze the data.Results(1)The relative expression levels of GRP78 protein in groups A,B and C were 1.33±0.46,0.93±0.34,0.64±0.30,respectively,the difference among groups was statistically significant(F=36.17,P＜0.05).The relative expression level of GRP78 protein in group A was higher than that in group B and group C(both P＜0.01),and the relative expression level of GRP78 protein in group B was higher than that in group C(P=0.0013).The relative expression levels of CHOP protein in the three groups were 1.29±0.38 in group A,0.90±0.34 in group B,and 0.59±0.29 in group C,the difference among groups was statistically significant(F=41.27,P＜0.05).The relative expression level of CHOP protein in group A was higher than that in group B and group C(both P＜0.01),and the relative expression level of CHOP protein in group B was higher than that in group C(P=0.0 004).(2)The relative expression levels of GRP78 mRNA in groups A,B,and C were 18.3±3.4,11.3±1.8,5.4±2.2,respectively,the difference among groups was statistically significant(F=189.20,P＜0.05).The relative expression level of GRP78 mRNA in group A was higher than that in group B and group C(both P＜0.01),and the relative expression level of GRP78 mRNA in group B was higher than that in group C(P＜0.01).The relative expression levels of CHOP mRNA in the three groups were 20.4±3.8 in group A,14.2±2.1 in group B,and 7.8±1.3 in group C,the difference among groups was statistically significant(F=171.80,P＜0.05).The relative expression level of CHOP mRNA in group A was higher than that in group B and group C(both P＜0.01),and the relative expression level of CHOP mRNA in group B was higher than that in group C(P＜0.01).Conclusion Low fluid shear stress may increase the protein and mRNA expression levels of GRP78 and CHOP in HUVECs.

Objective To investigate the effects of β-caryophyllene(BCP)on the browning of white adipose tissue in obese mice and the related mechanisms.Methods An obese mouse model was established via intraperitoneal injection of a high-fat diet supplemented with propylthiouracil saline solution[14.4 mg/(kg·d)]in male Kun-ming mice.Obesity model mice were randomly divided into a model group(Model group)and a BCP administra-tion group(BCP-50 group);normal diet mice were set up as a control group(Control group),with 8 mice in each group.BCP administration was given by gavage at a dose of 50 mg/kg once in the morning and once in the evening in the BCP-administered group,while the rest of the group was administered by gavage with aqueous solution of Tween 80 for 4 weeks.The oral glucose tolerance test was performed at the end of 4-week administration,and mice were executed after overnight fasting at the end of the experiment,and blood samples and adipose tissues were rap-idly collected for subsequent experimental tests.The kit was used to detect serological-related indexes;hematoxy-lin-eosin staining was conducted to observe the morphology of adipose tissue;immunohistochemical staining was carried out to observe the expression of uncoupling protein 1(UCP1)in adipose tissue;Western blot was employed to detect expression of peroxisome proliferator-activated receptor γ coactivator1-α(PGC1α),peroxisome prolifera-tor-activated receptor γ(PPARγ),UCP1 and cannabinoid receptor 2(CNR2)proteins in epididymal white adi-pose(eWAT).Results Compared with the model group,the body mass of obese mice in the BCP-50 group was significantly reduced(P＜0.05),food intake was decreased(P＜0.01),insulin resistance was improved(P＜0.000 1),and the serum content of low-density lipoprotein cholesterol(LDL-C)and nonesterified fatty acid(NE-FA)in the obese mice was significantly reduced(P＜0.000 1 and P＜0.01).Total cholesterol(TC),triglycer-ide(TG),and high-density lipoprotein cholesterol(HDL-C)contents did not change significantly.In addition,the adiposity coefficient and eWAT specific gravity of obese mice in the BCP-50 group were significantly decreased(P＜0.05);the adipocytes in eWAT and BAT were reduced;and the expression of the UCP1 protein was signifi-cantly elevated(P＜0.01 and P＜0.05).In addition to UCP1,the expression levels of PGC1α,PPARγ,and CNR2 proteins in the eWAT of obese mice in the BCP-50 group were also significantly elevated(P＜0.01,P＜0.05,and P＜0.001).Conclusion β-caryophyllene promotes white adipose tissue browning through up-regula-ting PPARγ/PGC-1α/UCP1 pathway expression,thus improving obesity.

Objective To investigate the effects of β-caryophyllene(BCP)on the browning of white adipose tissue in obese mice and the related mechanisms.Methods An obese mouse model was established via intraperitoneal injection of a high-fat diet supplemented with propylthiouracil saline solution[14.4 mg/(kg·d)]in male Kun-ming mice.Obesity model mice were randomly divided into a model group(Model group)and a BCP administra-tion group(BCP-50 group);normal diet mice were set up as a control group(Control group),with 8 mice in each group.BCP administration was given by gavage at a dose of 50 mg/kg once in the morning and once in the evening in the BCP-administered group,while the rest of the group was administered by gavage with aqueous solution of Tween 80 for 4 weeks.The oral glucose tolerance test was performed at the end of 4-week administration,and mice were executed after overnight fasting at the end of the experiment,and blood samples and adipose tissues were rap-idly collected for subsequent experimental tests.The kit was used to detect serological-related indexes;hematoxy-lin-eosin staining was conducted to observe the morphology of adipose tissue;immunohistochemical staining was carried out to observe the expression of uncoupling protein 1(UCP1)in adipose tissue;Western blot was employed to detect expression of peroxisome proliferator-activated receptor γ coactivator1-α(PGC1α),peroxisome prolifera-tor-activated receptor γ(PPARγ),UCP1 and cannabinoid receptor 2(CNR2)proteins in epididymal white adi-pose(eWAT).Results Compared with the model group,the body mass of obese mice in the BCP-50 group was significantly reduced(P＜0.05),food intake was decreased(P＜0.01),insulin resistance was improved(P＜0.000 1),and the serum content of low-density lipoprotein cholesterol(LDL-C)and nonesterified fatty acid(NE-FA)in the obese mice was significantly reduced(P＜0.000 1 and P＜0.01).Total cholesterol(TC),triglycer-ide(TG),and high-density lipoprotein cholesterol(HDL-C)contents did not change significantly.In addition,the adiposity coefficient and eWAT specific gravity of obese mice in the BCP-50 group were significantly decreased(P＜0.05);the adipocytes in eWAT and BAT were reduced;and the expression of the UCP1 protein was signifi-cantly elevated(P＜0.01 and P＜0.05).In addition to UCP1,the expression levels of PGC1α,PPARγ,and CNR2 proteins in the eWAT of obese mice in the BCP-50 group were also significantly elevated(P＜0.01,P＜0.05,and P＜0.001).Conclusion β-caryophyllene promotes white adipose tissue browning through up-regula-ting PPARγ/PGC-1α/UCP1 pathway expression,thus improving obesity.

Objective To investigate the effects of β-caryophyllene(BCP)on the browning of white adipose tissue in obese mice and the related mechanisms.Methods An obese mouse model was established via intraperitoneal injection of a high-fat diet supplemented with propylthiouracil saline solution[14.4 mg/(kg·d)]in male Kun-ming mice.Obesity model mice were randomly divided into a model group(Model group)and a BCP administra-tion group(BCP-50 group);normal diet mice were set up as a control group(Control group),with 8 mice in each group.BCP administration was given by gavage at a dose of 50 mg/kg once in the morning and once in the evening in the BCP-administered group,while the rest of the group was administered by gavage with aqueous solution of Tween 80 for 4 weeks.The oral glucose tolerance test was performed at the end of 4-week administration,and mice were executed after overnight fasting at the end of the experiment,and blood samples and adipose tissues were rap-idly collected for subsequent experimental tests.The kit was used to detect serological-related indexes;hematoxy-lin-eosin staining was conducted to observe the morphology of adipose tissue;immunohistochemical staining was carried out to observe the expression of uncoupling protein 1(UCP1)in adipose tissue;Western blot was employed to detect expression of peroxisome proliferator-activated receptor γ coactivator1-α(PGC1α),peroxisome prolifera-tor-activated receptor γ(PPARγ),UCP1 and cannabinoid receptor 2(CNR2)proteins in epididymal white adi-pose(eWAT).Results Compared with the model group,the body mass of obese mice in the BCP-50 group was significantly reduced(P＜0.05),food intake was decreased(P＜0.01),insulin resistance was improved(P＜0.000 1),and the serum content of low-density lipoprotein cholesterol(LDL-C)and nonesterified fatty acid(NE-FA)in the obese mice was significantly reduced(P＜0.000 1 and P＜0.01).Total cholesterol(TC),triglycer-ide(TG),and high-density lipoprotein cholesterol(HDL-C)contents did not change significantly.In addition,the adiposity coefficient and eWAT specific gravity of obese mice in the BCP-50 group were significantly decreased(P＜0.05);the adipocytes in eWAT and BAT were reduced;and the expression of the UCP1 protein was signifi-cantly elevated(P＜0.01 and P＜0.05).In addition to UCP1,the expression levels of PGC1α,PPARγ,and CNR2 proteins in the eWAT of obese mice in the BCP-50 group were also significantly elevated(P＜0.01,P＜0.05,and P＜0.001).Conclusion β-caryophyllene promotes white adipose tissue browning through up-regula-ting PPARγ/PGC-1α/UCP1 pathway expression,thus improving obesity.

Objective:The aim of this study was to explore whether hydroxysafflor yellow A (HSYA) combined with hyaluronidase (HAase) can enhance the therapeutic effect of arterial embolism caused by hyaluronic acid (HA) .Methods:Thirty-two white male rabbits were randomly divided into four groups, with 8 rabbits in each group, of which group A, B and C were experimental groups and group D was group control. An axial rectangular composite tissue flap sized 2.0 cm × 5.0 cm, with 1.0 cm pedicle width, and 4.0 cm from the root, was designed with the central auricular artery as the long axis on the dorsal side of the ear. The depth of incision reached the ventral perichondrium of the ear, and the flap was sutured continuously in situ and divided into three equal parts (area Ⅰ, Ⅱ, Ⅲ) from the proximal area to the distal area. The proximal end 1 cm to the flap and the central artery was the intersection point, into which 50 μl HA was injected, by which the model of HA arterial embolism was established. Each group was treated after 60 min. Group A: 20 ml solution HSYA was injected slowly into the thigh saphenous vein (the dosage of HSYA is calculated at 10 mg/kg) . Group B: 0.5 ml solution HAase was injected into the central auricular artery (400 U/ml) . Group C: 0.5 ml solution HAase with the same dosage of group B was injected into the central auricular artery and 20 ml solution HSYA with the same dosage of group A was injected slowly into the thigh saphenous vein. Group D and other parts of group A and B were injected with the same dosage of normal saline (NS) . The thigh saphenous veins of all groups were injected with the same dosage of solution once a day for 14 days. Flaps were observed immediately, 1, 7 and 14 days after establishment of hyaluronic acid arterial embolism models of tissue flaps, and dorsal and backlight auricular photographs were taken. On the postoperative 14th day, percentages of survival areas of the flaps were calculated, and samples were taken from areas II of tissue flaps, which were stained by hematoxylin-eosin (HE) and Masson, and were detected the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) . The measurement data conformed to normal distribution was represented as Mean ± SD. Single factor analysis of variance (ANOVA) was used to compare the differences among groups, and head-to-head comparison by LSD test. P <0.05 was considered statistically significant. Results:Tissue flaps of all groups were pale immediately after operation. On the first day after operation, the dark ischemic area appeared at the distal end of each group. On the postoperative 7th day, the ischemic area of each group was necrotic and blackened to varying degrees, and the non-necrotic area swelled obviously. On the postoperative 14th day, the ischemic area of each group was further necrotic, blackened, curled and the boundary was clear. Group C was the best, group D was the worst, and both group A and B were between the two. The swelling of non-necrotic areas in group A and C were basically reduced. HE staining showed that numerous thrombi and inflammatory cells infiltration were formed in group D, and group B was behind it, and thrombi were rare in group A and C. Masson staining showed that collagen fibers were arranged regularly in group C, and abundant collagen fibers were disintegrated and disordered in group D, and both group A and B were between the two. The percentages of survival areas of the flaps in group A, B, C and D were as follows: (69.87 ± 5.04) %, (85.03 ± 6.58) %, (93.93 ± 4.25) % and (49.22±9.64) %. There were statistical differences in pairwise comparison between groups (all P <0.05) . SOD activity of group A, B, C and D were as follows: (49.83±8.08) , (36.65±5.49) , (55.61±7.93) and (22.45 ± 5.47) U/mg prot. Except that group A vs. C, there were statistical differences between groups (all P <0.05) . MDA content of group A, B, C and D were as follows: (0.77±0.17) , (1.03±0.16) , (0.68±0.12) , and (0.41±0.09) nmol/mg prot. Except that group A vs. C, there were statistical differences between groups (all P <0.05) . Conclusions:Under the condition of animal experiment, compared with HAase, HSYA combined with HAase can significantly enhance the therapeutic effect of HA arterial embolism and increase the proportion of survival area of tissue flap.

Objective:To explore the therapeutic effect of hydroxysafflor yellow A (HSYA) combined with hyaluronidase (HAase) for arterial embolism caused by hyaluronic acid (HA).Methods:Thirty-two white male rabbits were randomly divided into four groups, with 8 rabbits in each group. Groups A, B and C were experimental groups, while group D served as the control group. An axial rectangular composite tissue flap sized 2.0 cm × 5.0 cm, with a pedicle width of 1.0 cm, and located 4.0 cm from the root, was designed with the central auricular artery as the long axis on the dorsal side of the ear. The incision depth reached the ventral perichondrium of the ear, and the flap was sutured continuously in place and divided into three equal parts (areas Ⅰ, Ⅱ, Ⅲ) from the proximal to the distal area. The proximal end, located 1 cm from the flap, and the central artery was the intersection point, where 50 μl of HA was injected to establish the model of HA arterial embolism. Each group was treated after 60 minutes. Group A: 20 ml of HSYA solution was slowly injected into the saphenous vein of the thigh (the dosage of HSYA was calculated at 10 mg/kg). Group B: 0.5 ml of HAase solution was injected into the central auricular artery (400 U/ml). Group C: 0.5 ml of HAase solution with the same dosage as in group B was injected into the central auricular artery, while 20 ml of HSYA solution with the same dosage as in group A was slowly injected into the saphenous vein. Group D and other parts of groups A and B were injected with the same dosage of normal saline (NS). The thigh saphenous veins of all groups were injected with the same dosage of solution once daily for 14 days. Flaps were observed immediately, 1, 7 and 14 days after establishing hyaluronic acid arterial embolism models of tissue flaps. Dorsal and backlight auricular photographs were taken. On the 14th day postoperatively, the survival areas of the flaps were calculated. Samples were taken from areas Ⅱof tissue flaps, stained with hematoxylin-eosin (HE) and Masson, to detected the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA). The measurement data that conformed to a normal distribution was represented as Mean ± SD. Single-factor analysis of variance (ANOVA) was used to compare the differences among groups, followed by head-to-head comparison using the LSD test. P<0.05 was considered statistically significant. Results:Tissue flaps from all groups appeared pale immediately after the operation. On the first day after the operation, a dark ischemic area appeared at the distal end of each group. On the 7th day postoperatively, the ischemic area of each group showed varying degrees of necrosis and blackening, while the non-necrotic area exhibited significant swelling. On the 14th day post-operation, the ischemic area in each group showed further necrosis, blackening, and curling, with clear boundaries. Group C was the best, group D was the worst, and both group A and B were in between the two. The swelling of non-necrotic areas in groups A and C was reduced. HE staining revealed numerous thrombi and infiltration of inflammatory cells in group D, with group B following closely behind. Thrombi were rare in groups A and C. Masson staining showed that collagen fibers were organized regularly in group C, while abundant collagen fibers were disintegrated and disordered in group D. Groups A and B exhibited characteristics that fell between the other two groups. The percentages of survival areas of the flaps in groups A, B, C and D were as follows: (69.87±5.04)%, (85.03±6.58)%, (93.93±4.25)% and (49.22±9.64)%. There were statistical differences in pairwise comparisons between groups (all P<0.05). SOD activity of groups A, B, C, and D were as follows: (49.83±8.08), (36.65±5.49), (55.61±7.93) and (22.45±5.47) U/mg prot. Except for the group A vs. C, there were statistical differences between the groups (all P<0.01). The MDA content of groups A, B, C and D were as follows: (0.77±0.17), (1.03±0.16), (0.68±0.12), and (0.41±0.09) nmol/mg prot. Except that group A vs. C, there were statistical differences between groups (all P<0.01). Conclusion:In animal experiments, it was found that compared to HAase alone, the combination of HSYA with HAase significantly improves the therapeutic outcomes of HA arterial embolism and increases the proportion of tissue flap survival area.

Objective:The aim of this study was to explore whether hydroxysafflor yellow A (HSYA) combined with hyaluronidase (HAase) can enhance the therapeutic effect of arterial embolism caused by hyaluronic acid (HA) .Methods:Thirty-two white male rabbits were randomly divided into four groups, with 8 rabbits in each group, of which group A, B and C were experimental groups and group D was group control. An axial rectangular composite tissue flap sized 2.0 cm × 5.0 cm, with 1.0 cm pedicle width, and 4.0 cm from the root, was designed with the central auricular artery as the long axis on the dorsal side of the ear. The depth of incision reached the ventral perichondrium of the ear, and the flap was sutured continuously in situ and divided into three equal parts (area Ⅰ, Ⅱ, Ⅲ) from the proximal area to the distal area. The proximal end 1 cm to the flap and the central artery was the intersection point, into which 50 μl HA was injected, by which the model of HA arterial embolism was established. Each group was treated after 60 min. Group A: 20 ml solution HSYA was injected slowly into the thigh saphenous vein (the dosage of HSYA is calculated at 10 mg/kg) . Group B: 0.5 ml solution HAase was injected into the central auricular artery (400 U/ml) . Group C: 0.5 ml solution HAase with the same dosage of group B was injected into the central auricular artery and 20 ml solution HSYA with the same dosage of group A was injected slowly into the thigh saphenous vein. Group D and other parts of group A and B were injected with the same dosage of normal saline (NS) . The thigh saphenous veins of all groups were injected with the same dosage of solution once a day for 14 days. Flaps were observed immediately, 1, 7 and 14 days after establishment of hyaluronic acid arterial embolism models of tissue flaps, and dorsal and backlight auricular photographs were taken. On the postoperative 14th day, percentages of survival areas of the flaps were calculated, and samples were taken from areas II of tissue flaps, which were stained by hematoxylin-eosin (HE) and Masson, and were detected the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) . The measurement data conformed to normal distribution was represented as Mean ± SD. Single factor analysis of variance (ANOVA) was used to compare the differences among groups, and head-to-head comparison by LSD test. P <0.05 was considered statistically significant. Results:Tissue flaps of all groups were pale immediately after operation. On the first day after operation, the dark ischemic area appeared at the distal end of each group. On the postoperative 7th day, the ischemic area of each group was necrotic and blackened to varying degrees, and the non-necrotic area swelled obviously. On the postoperative 14th day, the ischemic area of each group was further necrotic, blackened, curled and the boundary was clear. Group C was the best, group D was the worst, and both group A and B were between the two. The swelling of non-necrotic areas in group A and C were basically reduced. HE staining showed that numerous thrombi and inflammatory cells infiltration were formed in group D, and group B was behind it, and thrombi were rare in group A and C. Masson staining showed that collagen fibers were arranged regularly in group C, and abundant collagen fibers were disintegrated and disordered in group D, and both group A and B were between the two. The percentages of survival areas of the flaps in group A, B, C and D were as follows: (69.87 ± 5.04) %, (85.03 ± 6.58) %, (93.93 ± 4.25) % and (49.22±9.64) %. There were statistical differences in pairwise comparison between groups (all P <0.05) . SOD activity of group A, B, C and D were as follows: (49.83±8.08) , (36.65±5.49) , (55.61±7.93) and (22.45 ± 5.47) U/mg prot. Except that group A vs. C, there were statistical differences between groups (all P <0.05) . MDA content of group A, B, C and D were as follows: (0.77±0.17) , (1.03±0.16) , (0.68±0.12) , and (0.41±0.09) nmol/mg prot. Except that group A vs. C, there were statistical differences between groups (all P <0.05) . Conclusions:Under the condition of animal experiment, compared with HAase, HSYA combined with HAase can significantly enhance the therapeutic effect of HA arterial embolism and increase the proportion of survival area of tissue flap.

Objective:To explore the therapeutic effect of hydroxysafflor yellow A (HSYA) combined with hyaluronidase (HAase) for arterial embolism caused by hyaluronic acid (HA).Methods:Thirty-two white male rabbits were randomly divided into four groups, with 8 rabbits in each group. Groups A, B and C were experimental groups, while group D served as the control group. An axial rectangular composite tissue flap sized 2.0 cm × 5.0 cm, with a pedicle width of 1.0 cm, and located 4.0 cm from the root, was designed with the central auricular artery as the long axis on the dorsal side of the ear. The incision depth reached the ventral perichondrium of the ear, and the flap was sutured continuously in place and divided into three equal parts (areas Ⅰ, Ⅱ, Ⅲ) from the proximal to the distal area. The proximal end, located 1 cm from the flap, and the central artery was the intersection point, where 50 μl of HA was injected to establish the model of HA arterial embolism. Each group was treated after 60 minutes. Group A: 20 ml of HSYA solution was slowly injected into the saphenous vein of the thigh (the dosage of HSYA was calculated at 10 mg/kg). Group B: 0.5 ml of HAase solution was injected into the central auricular artery (400 U/ml). Group C: 0.5 ml of HAase solution with the same dosage as in group B was injected into the central auricular artery, while 20 ml of HSYA solution with the same dosage as in group A was slowly injected into the saphenous vein. Group D and other parts of groups A and B were injected with the same dosage of normal saline (NS). The thigh saphenous veins of all groups were injected with the same dosage of solution once daily for 14 days. Flaps were observed immediately, 1, 7 and 14 days after establishing hyaluronic acid arterial embolism models of tissue flaps. Dorsal and backlight auricular photographs were taken. On the 14th day postoperatively, the survival areas of the flaps were calculated. Samples were taken from areas Ⅱof tissue flaps, stained with hematoxylin-eosin (HE) and Masson, to detected the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA). The measurement data that conformed to a normal distribution was represented as Mean ± SD. Single-factor analysis of variance (ANOVA) was used to compare the differences among groups, followed by head-to-head comparison using the LSD test. P<0.05 was considered statistically significant. Results:Tissue flaps from all groups appeared pale immediately after the operation. On the first day after the operation, a dark ischemic area appeared at the distal end of each group. On the 7th day postoperatively, the ischemic area of each group showed varying degrees of necrosis and blackening, while the non-necrotic area exhibited significant swelling. On the 14th day post-operation, the ischemic area in each group showed further necrosis, blackening, and curling, with clear boundaries. Group C was the best, group D was the worst, and both group A and B were in between the two. The swelling of non-necrotic areas in groups A and C was reduced. HE staining revealed numerous thrombi and infiltration of inflammatory cells in group D, with group B following closely behind. Thrombi were rare in groups A and C. Masson staining showed that collagen fibers were organized regularly in group C, while abundant collagen fibers were disintegrated and disordered in group D. Groups A and B exhibited characteristics that fell between the other two groups. The percentages of survival areas of the flaps in groups A, B, C and D were as follows: (69.87±5.04)%, (85.03±6.58)%, (93.93±4.25)% and (49.22±9.64)%. There were statistical differences in pairwise comparisons between groups (all P<0.05). SOD activity of groups A, B, C, and D were as follows: (49.83±8.08), (36.65±5.49), (55.61±7.93) and (22.45±5.47) U/mg prot. Except for the group A vs. C, there were statistical differences between the groups (all P<0.01). The MDA content of groups A, B, C and D were as follows: (0.77±0.17), (1.03±0.16), (0.68±0.12), and (0.41±0.09) nmol/mg prot. Except that group A vs. C, there were statistical differences between groups (all P<0.01). Conclusion:In animal experiments, it was found that compared to HAase alone, the combination of HSYA with HAase significantly improves the therapeutic outcomes of HA arterial embolism and increases the proportion of tissue flap survival area.