Objective To explore the changes in macrophage infiltration behavior during the dynamic evolution of thrombi following the formation of acute carotid artery thrombosis occlusion(ACTO).Methods 15 healthy male New Zealand rabbits were selected to establish an ACTO model by causing injury to the rabbit carotid artery using surgical sutures treated with ferric chloride.All rabbits were randomly divided into 5 groups according to the end-point time using the random number table method,namely 24-hour group,1 week group,4week group,8 week group,and 12week group postoperatively,with 3 rabbits in each group.At 24 hours post-operation,the ACTO model was examined by DS A.At 24 hours,1 week,4 weeks,8 weeks,and 12 weeks post-operation,samples were taken from the thrombotic arterial segment of the 3 rabbits in each group and embedded in paraffin.The thrombus samples were stained with hematoxylin-eosin(HE)and Martius scarlet blue(MSB)to analyze changes in thrombus morphology and composition(including red blood cells,fibrin and collagen fibers).Orbit Imaging Analysis software was used for semi-quantitative analysis of the thrombus composition components.Using immunohistochemistry to detect the distribution of MO and M2 macrophages in thrombi,aimed to summarize the dynamic evolution of thrombus morphology,composition,and macrophage infiltration behavior at different stages following ACTO occurrence.Results The 24-hour DSA results indicated that all experimental rabbits successfully established the ACTO model.(1)HE staining showed a continuous increase in thrombus size from 24 hours to 1 week.By 4 weeks,signs of thrombus dissolution appeared,and at 8 weeks,neovascularization was observed within the thrombus.By 12 weeks,signs of fibrosis were evident in the thrombus.(2)MSB staining revealed that during the acute phase of thrombus formation(within 24 hours after surgery),red blood cells were the predominant component initially,but after this period,fibrin and collagen fibers became the main components.(3)The detection results of MO macrophages showed that 24 hours after surgery,MO macrophages in the thrombus were not evenly distributed throughout the thrombus,but mainly gathered at the thrombus edge;at 1 week after surgery,the positive area percentage of MO macrophage in the thrombus increased compared with 24 hours after surgery(thrombus edge:[41.7±27.0]％vs.[24.6±16.7]％,thrombus core:[35.7±19.6]％vs.[11.1±10.4]％,all P＜0.001),and evenly distributed within the thrombus;at 4 weeks after surgery,MO macrophages in the thrombus decreased compared with 1 week after surgery(thrombosis edge:[10.7±6.1]％vs.[41.7±27.0]％,thrombus core:[12.1±8.5]％vs.[35.7±19.6]％,all P＜0.001),the differences were statistically significant.At 4,8,and 12 weeks after surgery,MO macrophages within the thrombus did not change significantly with time(thrombus edge:[10.7±6.1]％,[8.0±7.7]％,and[8.9±5.3]％;thrombus core:[12.1±8.5]％,[9.5±4.2]％,and[15.7±11.0]％),and the differences were not statistically significant(all P＞0.05).In addition,at 12 weeks after surgery,MO macrophages at the thrombus edge was less than the thrombus core([8.9+5.3]％vs.[15.7±11.0]％,P＜0.01).The detection results of M2 macrophages showed that 24 hours after surgery,M2 macrophages in the thrombus were widely distributed throughout the thrombus;at 1 week after surgery,the positive area percentage of M2 macrophages in the thrombus increased compared with 24 hours after surgery(thrombus edge:[22.1±11.3]％vs.[11.4±8.7]％,P＜0.001;thrombus core:[24.5±9.8]％vs.[7.6±6.0]％,P＜0.001);at 4 weeks after surgery,M2 macrophage in the thrombus decreased compared with 1 week after surgery(thrombosis edge:[10.6±3.7]％vs.[22.1±11.3]％,P＜0.001;thrombus core:[9.2±4.3]％vs.[24.5±9.8]％,P＜0.001);at 8 weeks after surgery,M2 macrophages in the thrombus increased compared with 4 weeks after surgery([17.9±8.8]％vs.[9.2±4.3]％,P＜0.001),and the differences were statistically significant.However,M2 macrophages in the thrombus did not change significantly from 8 weeks to 12 weeks after surgery(thrombus edge:[9.4±6.3]％vs.[8.5±5.3]％,P＞0.05;thrombus core:[17.9±8.8]％vs.[14.4±10.0]％,P＞0.05).In addition,at 8 and 12 weeks after surgery,M2 macrophages in the thrombus core was greater than the thrombus edge(8 weeks after surgery:[17.9±8.8]％vs.[9.4±6.3]％,P＜0.001;12weeks after surgery:[14.4±10.0]％vs.[8.5±5.3]％,P＜0.001).Conclusions This study successfully established an ACTO animal model and demonstrated for the first time the dynamic evolution of macrophages within 12 weeks post-thrombus formation.Macrophages may played a significant role in both thrombus formation and fibrinolysis,as well as in the promotion of thrombus dissolution and the formation of new blood vessels within the thrombus which may potentially promote the spontaneous reperfusion of the occluded vessels.The results of this study need further verification.

Objective:To analyze the clinical phenotype, sperm morphological characteristics and assisted reproductive therapy efficiency in patients with total globozoospermia.Methods:Four male patients with total globozoospermia were collected during November 2019 to May 2022 from Reproductive Medical and Genetic Center, the People's Hospital of Guangxi Zhuang Autonomous Region. Peripheral blood samples were collected for genetic detection and the whole exome sequencing to explore the pathogenic genes. Semen characteristics, sperm morphology and ultrastructure were analyzed. Four patients were treated with intracytoplasmic sperm injection (ICSI) combined with artificial oocyte activation (AOA). Conventional ICSI cycles ( n=9) were selected as control group, and the development dynamic parameters were monitored by Time-lapse. The fertilization and embryo development parameters, developmental dynamic parameters and clinical outcomes were analyzed between the two groups. Results:Four patients were complicated with low sperm motility and increased sperm DNA fragmentation. Sperm morphology analysis and acrosome fluorescence staining represented that all the spermatozoas were with a small round head lacked of acrosome. By the transmission electron microscope, it was observed that round-headed spermatozoas were lacked of acrosome completely, loose chromatin structure, vacuolation and other abnormal changes in the head, mitochondrial sheath in neck were reduced arranged in disorder, and the structure of "9+2" of the flagellar axial filament was incomplete. Of the 4 patients, 1 was homozygous deletion of DPY19L2 gene and 1 was homozygous frameshift mutation of DPY19L2 gene. There were no significance differences in fertilization rate, two pronuclei fertilization rate, day 3 high-quality embryo rate, day 6 blastocyst formation rate and day 6 high-quality blastocyst formation rate between total globozoospermia group and control group (all P>0.05). The developmental dynamic parameters as the time at which the second polar body is extruded, the time when both (or the last) PN disappear, two to six discrete cells in the total globozoospermia group were significantly earlier than those in control group, and the difference was statistically significant (all P<0.05). There was no significant difference in the embryo cleavage patterns between the two groups ( P>0.05). Among the 4 patients with total globozoospermia, 2 live births with signal healthy male baby were achieved by fresh embryo transfer, and 2 live births with one signal healthy male baby and one healthy female baby were achieved by frozen-thawed embryo transfer respectively. Conclusion:Abnormal morphology characteristics of spermatozoas from patients with total globozoospermia are obvious, patients with total globozoospermia could have favorable clinical outcomes following ICSI combined with AOA.

Objective:To analyze the clinical phenotype, sperm morphological characteristics and assisted reproductive therapy efficiency in patients with total globozoospermia.Methods:Four male patients with total globozoospermia were collected during November 2019 to May 2022 from Reproductive Medical and Genetic Center, the People's Hospital of Guangxi Zhuang Autonomous Region. Peripheral blood samples were collected for genetic detection and the whole exome sequencing to explore the pathogenic genes. Semen characteristics, sperm morphology and ultrastructure were analyzed. Four patients were treated with intracytoplasmic sperm injection (ICSI) combined with artificial oocyte activation (AOA). Conventional ICSI cycles ( n=9) were selected as control group, and the development dynamic parameters were monitored by Time-lapse. The fertilization and embryo development parameters, developmental dynamic parameters and clinical outcomes were analyzed between the two groups. Results:Four patients were complicated with low sperm motility and increased sperm DNA fragmentation. Sperm morphology analysis and acrosome fluorescence staining represented that all the spermatozoas were with a small round head lacked of acrosome. By the transmission electron microscope, it was observed that round-headed spermatozoas were lacked of acrosome completely, loose chromatin structure, vacuolation and other abnormal changes in the head, mitochondrial sheath in neck were reduced arranged in disorder, and the structure of "9+2" of the flagellar axial filament was incomplete. Of the 4 patients, 1 was homozygous deletion of DPY19L2 gene and 1 was homozygous frameshift mutation of DPY19L2 gene. There were no significance differences in fertilization rate, two pronuclei fertilization rate, day 3 high-quality embryo rate, day 6 blastocyst formation rate and day 6 high-quality blastocyst formation rate between total globozoospermia group and control group (all P>0.05). The developmental dynamic parameters as the time at which the second polar body is extruded, the time when both (or the last) PN disappear, two to six discrete cells in the total globozoospermia group were significantly earlier than those in control group, and the difference was statistically significant (all P<0.05). There was no significant difference in the embryo cleavage patterns between the two groups ( P>0.05). Among the 4 patients with total globozoospermia, 2 live births with signal healthy male baby were achieved by fresh embryo transfer, and 2 live births with one signal healthy male baby and one healthy female baby were achieved by frozen-thawed embryo transfer respectively. Conclusion:Abnormal morphology characteristics of spermatozoas from patients with total globozoospermia are obvious, patients with total globozoospermia could have favorable clinical outcomes following ICSI combined with AOA.

OBJECTIVE: To isolate the phenolic amides from the dried bulbs of Allium chinense and investigate their myocardium protective activities. METHODS: The chemical constituents were isolated and purified by combining with silica gel column, Sephadex LH-20 column, HPLC and other chromatography techniques. Their structures were elucidated by NMR techniques and mass spectrometry. The isolated compounds were evaluated to determine their protective effect for myocardium cells in vitro. RESULTS: Two new phenolic amides, namely, alichinemide I ( 1) and alichinemide II ( 2), and six konwn amides were isolated from the dried bulbs of A. chinense. The structures of compounds 3- 8 were identified as 3-indolcarbaldehyde ( 3), 1-(2-aminophenyl)urea ( 4), 2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylic acid ( 5), N-trans-feruloyltyramine ( 6), N-trans-p-coumaroyltyramine ( 7), and N-(3,4-dimethoxyphenethyl) acetamide ( 8). Compound 3 (50 μmol/L) showed significant inhibitory effect on the damage of H9c2 myocardial cells induced by H₂O₂in vitro. CONCLUSION Compounds 1 and 2 were new phenolic amides. Compound 3 could be one of the potential myocardium protective constituents of A. chinense.

Objective The chemical composition and blood components of Tianmaxingnao capsules were discovered and examined using UPLC-Q/TOF-MS,and the possible pharmacological substance basis was preliminarily elucidated.Methods An UPLC-Q/TOF-MS method was developed in this study to determine the chemical composition of Tianma Xingnao capsules.After administration of Tianmaxingnao capsules,gather and examine rat plasma samples to investigate the exposed components of Tianmaxingnao capsules in rats.Results A total of 195 chemical components were identified in Tianmaxingnao capsules,including phenols,triterpenoid saponins,phenylethanol glycosides,cyclic ether terpenes,lipids,and phenylpropanoids.These components include those that are typical of Gastrodia elata Bl,Pheretima,Cistanche deserticola Ma,Rehmannia glutinosa,Polygala hybrida DC,and Acorus tatarinowii.Rat plasma samples were used to identify 37 prototype components and 3 metabolites of Tianmaxingnao capsules after they were administered.Conclusion This approach is easy to use,effective,sensitive,and precise.It may be used to investigate Tianmaxingnao capsules and the components that reach the bloodstream in detail,as well as to provide a first understanding of the pharmacological basis of the capsules.This study serves as a foundation for clarifying the pharmacological basis of Tianmaxingnao capsules and holds some reference value in exposing the pharmacological mechanism of the product.

Objective To study the chemical constituents from ethyl acetate extracts of the strips of Semiliquidambar cathayensis.Methods The strips of S.cathayensis were extracted by 80%ethanol and the extracts were evaporated.Fourteen compounds in ethyl acetate extracts were isolated and purified by various chromatographic techniques,such as silica gel,Sephadex LH-20 column and pre-HPLC,etc.Their structures were identified on the basis of physicochemical properties and spectroscopic analysis.Antioxidant activity test was used to evaluate total extraction,each extraction part and the isolates.Results Fourteen compounds were isolated from the strips of S.cathayensis and identified by NMR as tetradecanoic acid(1),stearic acid(2),sesamin(3),9-octadecenoic acid(4),linoleic acid(5),dibutylphthalate(6),stigmasterol(7),β-sitosterol(8),lupeol(9),oleanolic acid(10),lup-20(29)-ene-3-[3-keto-hexadecanoate](11),peujaponisin(12),C-veratroylglycol(13),and 2,3-dihydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone(14).Conclusion Compounds 1,3,4,5,6,7,9,11,12,13 and 14 were isolated from this plant for the first time.The EA part,compounds 13(C-veratroylglycol)and 14(2,3-dihydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone)showed significant antioxidative effects.

Objective:To investigate whether the early developmental dynamic phenotypes and kinetic parameters of embryos used for embryo selection are affected by the differences in the components of culture media utilized.Methods:The clinical data of patients undergoing in vitro fertilization (IVF) at Center for Reproductive Medicine and Genetics of People's Hospital of Guangxi Zhuang Autonomous Region from October 2016 to December 2018 were analyzed in a retrospective cohort study. According to the different culture media utilized, IVF cycles were divided into Cook group and Vitrolife group. After 1∶1 propensity-score matching (PSM), 59 IVF cycles were included in each group. Time-lapse imaging technology was used to analyze the early developmental dynamics of normal fertilized embryos between insemination and 68 h post insemination. Seven developmental dynamic phenotypes of embryos were annotated and the differences in the composition of dynamic phenotypes were compared between the two groups. Thirteen early developmental kinetic parameters were calculated, and the differences in the kinetic parameters of normal dynamic phenotypic embryos between the two groups were compared. According to the two published time-lapse embryo selection algorithms, the hierarchical distribution of normal dynamic phenotypic embryos of two groups was compared. Results:1) In Cook group, the composition of developmental dynamic phenotypes of embryos were 54.0% normal phenotype, 3.0% abnormal first cytokinesis (A1 cyt), 17.4% abnormal cleavage (AC), 5.2% reverse cleavage (RC), 3.2% chaotic cleavage (CC), 3.5% multinucleation (Mn) and 13.7% mixed phenotype, which were 49.3%, 4.0%, 19.1%, 7.5%, 2.1%, 6.4% and 11.6% in Vitrolife group, respectively. No statistically significant differences were observed between the two groups for the composition of dynamic developmental phenotypes ( P>0.05). 2) Compared with Vitrolife group, the 13 developmental kinetic parameters (tPNa, tPNf, t2, t3, t4, t5, t6, t7, t8, cc2, s2, t5_PNf and t8_PNf) of normal dynamic phenotype embryos in Cook group were slightly longer, and the average number of blastomeres in 68 h post insemination (EB68hpi) was less, but the differences were not statistically significant (all P>0.05). 3) No significant difference in hierarchical distribution of embryos was observed between Vitrolife group and Cook group according to algorithm A ( P>0.05). The difference of embryo hierarchical distribution between the two groups was statistically significant according to algorithm B ( P=0.040), the proportion of grade A + embryos in Vitrolife group was higher than that in Cook group [59.8% (125/209) vs. 43.3% (94/217)], and grade C proportion was lower [9.6% (20/209) vs. 20.3% (44/217)]. Conclusion:Although the early developmental dynamic phenotypes and kinetic parameters of embryos were not affected by the differences between Cook and Vitrolife sequential culture media, the applicability of different time-lapse embryo selection algorithms to the culture media is different, the embryo culture media utilized should be considered when selecting or constructing the algorithms.

Objective:To investigate whether the early developmental dynamic phenotypes and kinetic parameters of embryos used for embryo selection are affected by the differences in the components of culture media utilized.Methods:The clinical data of patients undergoing in vitro fertilization (IVF) at Center for Reproductive Medicine and Genetics of People's Hospital of Guangxi Zhuang Autonomous Region from October 2016 to December 2018 were analyzed in a retrospective cohort study. According to the different culture media utilized, IVF cycles were divided into Cook group and Vitrolife group. After 1∶1 propensity-score matching (PSM), 59 IVF cycles were included in each group. Time-lapse imaging technology was used to analyze the early developmental dynamics of normal fertilized embryos between insemination and 68 h post insemination. Seven developmental dynamic phenotypes of embryos were annotated and the differences in the composition of dynamic phenotypes were compared between the two groups. Thirteen early developmental kinetic parameters were calculated, and the differences in the kinetic parameters of normal dynamic phenotypic embryos between the two groups were compared. According to the two published time-lapse embryo selection algorithms, the hierarchical distribution of normal dynamic phenotypic embryos of two groups was compared. Results:1) In Cook group, the composition of developmental dynamic phenotypes of embryos were 54.0% normal phenotype, 3.0% abnormal first cytokinesis (A1 cyt), 17.4% abnormal cleavage (AC), 5.2% reverse cleavage (RC), 3.2% chaotic cleavage (CC), 3.5% multinucleation (Mn) and 13.7% mixed phenotype, which were 49.3%, 4.0%, 19.1%, 7.5%, 2.1%, 6.4% and 11.6% in Vitrolife group, respectively. No statistically significant differences were observed between the two groups for the composition of dynamic developmental phenotypes ( P>0.05). 2) Compared with Vitrolife group, the 13 developmental kinetic parameters (tPNa, tPNf, t2, t3, t4, t5, t6, t7, t8, cc2, s2, t5_PNf and t8_PNf) of normal dynamic phenotype embryos in Cook group were slightly longer, and the average number of blastomeres in 68 h post insemination (EB68hpi) was less, but the differences were not statistically significant (all P>0.05). 3) No significant difference in hierarchical distribution of embryos was observed between Vitrolife group and Cook group according to algorithm A ( P>0.05). The difference of embryo hierarchical distribution between the two groups was statistically significant according to algorithm B ( P=0.040), the proportion of grade A + embryos in Vitrolife group was higher than that in Cook group [59.8% (125/209) vs. 43.3% (94/217)], and grade C proportion was lower [9.6% (20/209) vs. 20.3% (44/217)]. Conclusion:Although the early developmental dynamic phenotypes and kinetic parameters of embryos were not affected by the differences between Cook and Vitrolife sequential culture media, the applicability of different time-lapse embryo selection algorithms to the culture media is different, the embryo culture media utilized should be considered when selecting or constructing the algorithms.

Objective:To analyze the prognostic factors of breast cancer patients with isolated chest wall recurrence (ICWR) after mastectomy, and investigate the optimal treatment.Methods:A total of 201 breast cancer patients with ICWR after mastectomy who were treated in Cancer Hospital, Chinese Academy of Medical Sciences and the Fifth Medical Center Chinese PLA General Hospital from October 1998 to April 2018 were retrospectively analyzed. The median follow-up was 92.8 months and survival data were obtained.Results:Among 201 patients with ICWR, 103 patients developed subsequent locoregional recurrence (sLRR) and 5-year cumulative sLRR rate was 49.1%; 134 patients developed distant metastasis (DM) and 5-year DM rate was 64.4%; 103 patients died, the median progression-free survival (PFS) was 17.4 months and the 5-year PFS rate was 23.2%; the median overall survival (OS) was 62.5 months and the 5-year OS rate was 52.1%. Multivariate analysis showed that the recurrence interval ( HR=2.17, 95% CI: 1.26-3.73) and the locoregional treatment ( HR=1.59, 95% CI: 1.05-2.40) were the independent prognostic factors for sLRR. The initial HER2 status ( HR=1.60, 95% CI: 1.03-2.48) was the independent prognostic factor for DM. The recurrence interval ( HR=1.99, 95% CI: 1.30-3.04), the locoregional treatment ( HR=1.99, 95% CI: 1.43-2.76) and the treatment modalities after recurrence ( HR=1.70, 95% CI: 1.18-2.46) were the independent prognostic factors for PFS. The initial HER2 status ( HR=1.69, 95% CI: 1.02-2.81), the recurrence interval ( HR=1.85, 95% CI: 1.15-2.98) and the treatment modalities after recurrence ( HR=2.48, 95% CI: 1.56-3.96) were the independent prognostic factors for OS. Conclusions:Breast cancer patients after ICWR have an optimistic OS until now, but the risk of sLRR and DM is high. Comprehensive treatment modalities including surgery, radiotherapy and systemic therapy improve the outcome of breast cancer patients with ICWR after mastectomy.

Objective:To investigate the radiation field and dose selection of patients with isolated chest wall recurrence (ICWR) after modified radical mastectomy, and analyze the prognostic factors related to subsequent chest wall recurrence.Methods:Clinical data of 201 patients with ICWR after mastectomy admitted to the Fifth Medical Center, Chinese PLA General Hospital from 1998 to 2018 were retrospectively analyzed. None of the patients received postoperative adjuvant radiotherapy. After ICWR, 48 patients (73.6%) underwent surgery and 155 patients (77.1%) received radiotherapy. Kaplan-Meier method was used to calculate the post-recurrence progression-free survival (PFS) rates and the difference was compared by log-rank test. Multivariate analysis was performed using Cox regression model. Competing risk model was adopted to estimate the subsequent local recurrence (sLR) rates after ICWR and the difference was compared with Gray test. Multivariate analysis was conducted using F&G analysis. Results:With a median follow up of 92.8 months after ICWR, the 5-year PFS rate was 23.2%, and the 5-year sLR rate was 35.7%. Multivariate analysis showed that patients with surgery plus radiotherapy and recurrence interval o F>12 months had a lower sLR rate. Patients with recurrence interval o F>48 months, local plus systemic treatment and surgery plus radiotherapy had a higher PFS rate. Among the 155 patients who received chest wall radiotherapy after ICWR, total chest wall irradiation plus local boost could improve the 5-year PFS rate compared with total chest wall irradiation alone (34.0% vs. 15.4%, P=0.004). Chest wall radiation dose (≤60 Gy vs.>60 Gy) exerted no significant effect upon the sLR and PFS rates (both P>0.05). In the 53 patients without surgery, the 5-year PFS rates were 9.1% and 20.5%( P=0.061) with tumor bed dose ≤60 Gy and>60 Gy, respectively. Conclusions:Local radiotherapy is recommended for patients with ICWR after modified radical mastectomy of breast cancer, including total chest wall radiation plus local boost. The radiation dose for recurrence should be increased to 60 Gy, and it should be above 60 Gy for those who have not undergone surgical resection. In addition, patients with ICWR still have a high risk of sLR, and more effective treatments need to be explored.