Objective To investigate the temporal changes in pathological damage and macrophage polarization in liver and spleen tissues of mice infected with Angiostrongylus cantonensis, and to preliminarily unravel the peripheral immune responses during the early stage of A. cantonensis infection. Methods Forty female BALB/c mice at ages of 6 to 8 weeks were randomly divided into four groups, including the control group and 7-, 14-, and 21-day infection groups, with 10 mice in each group. Each mouse in the infection groups was inoculated with 30 third-stage (L3) larvae of A. cantonensis by oral gavage, and five mice were randomly selected from each infection group on days 7, 14, and 21 post-infection, while mice in the control group were given the same volume of physiological saline and five mice were randomly selected from the control group on the day of oral gavage. Mouse liver and spleen tissues were sampled. The histopathological changes of mouse liver and spleen tissues were observed using hematoxylin and eosin (HE) staining, and the percentage of positive staining area and the co-localization positive rates of the macrophage surface antigens F4/80, CD86, and CD206 were quantified in mouse liver and spleen tissues using immunohistochemical and immunofluorescence staining. In addition, five mice were collected from each infection group on days 7, 14, and 21 post-infection, and five mice were collected from the control group on the day of oral gavage. Mouse liver and spleen tissues were sampled for detection of macrophage markers CD86 and CD206 and macrophage phenotyping using flow cytometry, and the expression of M1 macrophage markers, including inducible nitric oxide synthase (Nos2), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and M2 markers, including arginase 1 (Arg1), mannose receptor C-type 1 (Mrc1) and chitinase-like protein 3 (Chil3) was quantified in mouse liver and spleen tissues using real-time quantitative PCR (RT-qPCR) assay. Results Proliferative lesions of the hepatocyte were observed in mouse liver tissues and the follicular structures of the mouse spleen white pulp were disrupted 21 days post-infection with A. cantonensis. Immunohistochemical staining showed that there were significant differences in the percentages of F4/80, CD86 and CD206 positive staining areas in the liver and spleen tissues among the four groups of mice (F = 242.40, 197.14, 183.19, 157.65, 242.35 and 146.24; all P values < 0.001), and the percentages of positive staining in the liver and spleen tissues of mice in the 14-day infection group [(4.45 ± 0.51)%, (3.74 ± 0.67)%, (8.32 ± 0.72)%, (16.56 ± 1.14)%, (11.62 ± 0.52)%, and (8.29 ± 0.72)%, respectively] and the 21-day infection group [(3.70 ± 0.11)%, (3.22 ± 0.43)%, (11.53 ± 1.03)%, (12.59 ± 1.05)%, (9.02 ± 0.83)%, and (11.67 ± 1.10)%, respectively] were higher than in the control group [(0.35 ± 0.16)%, (0.40 ± 0.02)%, (0.93 ± 0.05)%, (2.78 ± 0.26)%, (2.33 ± 0.20)%, and (1.85 ± 0.20)%, respectively] (all P values < 0.05). Immunofluorescence staining showed significant differences in the positive rates of F4/80 co-localization with CD86 and CD206 in mouse liver and spleen tissues among the four groups (F = 24.42, 25.28, 54.51 and 130.55; all P values < 0.001). Flow cytometry detected significant differences in the proportions of CD86⁺ and CD206⁺ macrophages in mouse liver and spleen tissues among the four groups (F = 67.98, 18.41, 29.77, 172.80; all P values < 0.001), and the proportions of CD206⁺ macrophages in the liver and spleen of the 21-day infection group were significantly higher than those in the control group [(9.25 ± 2.55)% vs (3.83 ± 0.72)%, and (4.22 ± 0.56)% vs (0.47 ± 0.18)%, respectively] (both P values < 0.05). In addition, RT-qPCR assay quantified significant differences in the relative mRNA expression of M1 macrophage markers (IL-1β, TNF-α and Nos2) and M2 macrophage markers (Arg1, Chil3 and Mrc1) in mouse liver and spleen tissues among the four groups (F = 41.30, 31.82, 199.33, 19.96, 62.01, 119.76, 23.67, 95.90, 72.27, 82.59, 123.41 and 29.75; all P values < 0.05). Conclusions A. cantonensis infection may cause progressive pathological damage in mouse liver and spleen tissues, accompanied by dynamic temporal changes in macrophage polarization. M1 macrophage polarization predominates at the early stage of A. cantonensis infection and shifts towards M2 polarization at the later stages, suggesting that M2 polarization may participate in immune regulation at late stages of A. cantonensis infection by suppressing excessive inflammatory responses and promoting tissue repair.

Aim To study the protective effect of the silibinin derivative Sil-1 on acute myocardial ischemia in SD rats and its mechanism of action.Methods Af-ter 18 hours of oxygen-glucose deprivation and treat-ment of H9c2 cells,the protective effect of Sil-1 on rat cardiomyocytes was examined.SD rats were treated 30 minutes before surgery,followed by 24 h ligation of the left anterior descending coronary artery.The cardiopro-tective effects of Sil-1 and its mechanisms for improving myocardial ischemic injury were investigated using pro-teomics technology.Results In vitro,compared with the control group,the activity of H9c2 cells in the mod-el group showed reduced cell viability,increased dead cells,elevated ROS and higher levels of LDH and in-flammatory cytokines TNF-α,IL-1β and IL-6 in the culture medium.Sil-1 could improve the above condi-tions to different degrees.In vivo,compared with the control group,rats in the model group showed signifi-cantly higher T waves on electrocardiogram,significant ischemic areas in the heart section,disorganized ar-rangement of cardiomyocytes,increased inflammatory factor infiltration and elevated CK,CK-MB,LDH and inflammatory factors TNF-α,IL-6 and IL-1β.Besides,NF-κB phosphorylation levels in myocardial tissue in-creased.Sil-1 improved the above conditions to varying degrees.The results of proteomics showed that 90 pro-teins were found between the control vs model group and the Sil-1 vs model group,and KEGG enrichment a-nalysis showed that MAPK,chemokines,VEGF and other signaling pathways were abundant.Western blot results showed that Sil-1 blocked the phosphorylation of ERK,JNK and p38 MAPK.Conclusions Sil-1 inhib-its the MAPK pathway by blocking the phosphorylation of JNK,ERK,and p38 MAPK,and achieves a protec-tive effect on rats with acute myocardial infarction.

A recombinant adenovirus(rAd)for expression of Mycobacterium tuberculosis(M.tb)c-di-AMP phosphodiesterase CnpB was constructed,and its induced humoral immune response was detected.The codon-optimized gene of M.tb CnpB was cloned into the adenoviral plasmid pcADV.The recombinant plasmid pcADV-CnpB was transfected into HEK293T cells,and expression was detected with Western blot.The recombinant plasmid pcADV-CnpB and the backbone plasmid were co-transfected into HEK293T cells to obtain the recombinant adenovirus rAd-CnpB.rAd-CnpB was amplified in HEK293T cells,and the target protein expression of rAd-CnpB was detected with Western blot and immunofluorescence.Mice were immunized with rAd-CnpB intranasally,and their sera and bronchoalveolar lavage fluid(BALF)were collected.ELISA was used to detect levels of antigen-specific antibodies.Restriction enzyme digestion and sequencing indicated that the recombinant plasmid pcADV-CnpB was successfully constructed and led to protein expression in eukaryotic cells.rAd-CnpB was packaged and produced in HEK293T cells.After amplification and purification,rAd-CnpB with a titer of 5.53×1010 PFU/mL was obtained.rAd-CnpB led to CnpB expression in HEK293T cells.Intranasal immunization with rAd-CnpB increased levels of IgG and secretory IgA in BALF and led to high levels of IgG in sera.rAd-CnpB,the recombinant adenovirus for expression of c-di-AMP phosphodiesterase CnpB was successfully constructed,and was found to induce antigen-specific humoral and mucosal immune responses through mucosal immunization.Thus,rAd-CnpB may be used in further research on new TB vaccine strategies.

This study assessed the role and mechanism of the recombinant Bacillus Calmette-Gue?rin vaccine(rBCG)with c-di-AMP adjuvant in regulating metabolism and immunity in epididymal white adipose(eWAT)in mice.Male C57BL/6 mice were intravenously immunized with BCG and rBCG,and their body weights were monitored.eWAT was isolated from the mice,and the stromal vascular fractions(SVFs)cell number was counted with a hemocytometer.Sections of mouse adipose tissue were prepared,and the size,number,and morphology of eWAT adipocytes and crown-like structure(CLS)formation were compared under a microscope after HE staining.The transcription levels of lipid metabolism-associated factors,cytokines and aging-associated genes in each group were determined with qRT-PCR.The body weights of mice gradually increased after immunization with BCG and rBCG.The proportions of eWAT increased,and the SVFs cell number decreased,in rBCG immunized mice.HE staining indicated that BCG immunization promoted hyperplasia,whereas rBCG immunization promoted hypertrophy of eWAT adipocytes;moreover,both BCG and rBCG immunization induced CLS formation in eWAT.The qRT-PCR results indicated that rBCG immunization inhibited the expression of genes associated with lipolysis and energy expenditure in eWAT.BCG immunization had little effect on cytokine transcription,whereas rBCG significantly induced the transcription of IFN-γ and IL-1Ra,and inhibited that of IL-15 and IL-2,but did not induce the expression of aging-associated genes.Thus,rBCG immunization induced eWAT adipocyte hypertrophy,which was associated with the inhibition of eWAT lipolysis and the regulation of cytokine expression.

Objective To evaluate the clinical outcomes of 514 cases of anterior cruciate ligament(ACL)reconstruction using hamstring tendon autograft and to observe postoperative changes and recovery of the grafts through second-look arthroscopy.Methods This retrospective study collected data from 514 patients who underwent ACL reconstruction with hamstring tendon autograft between May 2015 and June 2018,with a follow-up of at least one year.Knee function recovery and stability were assessed using the Lysholm score,International Knee Documentation Committee(IKDC)score,and Tegner score,along with the pivot shift test and Lachman test.During the second-look arthroscopy,key observations included the synovial coverage,continuity of the reconstructed ligament,and any intra-articular abnormalities.Results The time interval between ACL reconstruction and second-look arthroscopy ranged from 12 to 28 months,with an average of 20 months.Postoperative infection occurred in 2 cases,both of which were successfully treated with arthroscopic debridement and drainage.No other patients experienced infections,graft resorption,or other complications.At the second-look arthroscopy,the Lysholm score significantly improved from(43.56±9.89)preoperative to(92.21±6.12)postoperatively,the difference was statistically significant(P<0.05);The IKDC score increased from(20.32±7.87)to(85.67±10.43),the difference was statistically significant(P<0.05);The Tegner score improved from(4.31±0.82)to(6.61±1.21),the difference was statistically significant(P<0.05).Second-look arthroscopy revealed that the ligament remained intact in 375 patients,with partial tears in 139 patients,ligament tension was maintained in 447 patients,while 67 patients had laxity,the reconstructed ACL graft was deemed to be in good condition in 435 patients and suboptimal in 79 patients,there were significant differences observed in pre-and post-pivot shift test and Lachman test(P<0.05);Among the 514 patients,188(36.58%)successfully returned to sport.Conclusion ACL reconstruction using hamstring tendon autograft effectively restores knee function and stability.In patients followed for more than one years,the grafts show good vascularization and synovial coverage.Emphasis should be placed on systematic postoperative rehabilitation to optimize recovery.

Sepsis is a life-threatening organ dysfunction disease caused by a dysregulated host response to infection.It has com-plex pathophysiological changes,and in severe cases,it can rap-idly develop into septic shock and multiple organ dysfunction or multiple organ failure.At present,the pathological mechanism of sepsis and its induced organ dysfunction is complex and the in-fluencing factors are numerous.So far,there is still a lack of specific and effective treatment strategies.RNA modify-N6-methyladenosine(m6 A)is one of the most common post-tran-scriptional modifications on eukaryotic RNAs.It is involved in the regulation of the occurrence and development of a variety of inflammatory diseases,including sepsis,and even multiple organ dysfunction induced by sepsis by affecting the metabolism of RNAs.It includes cardiac dysfunction,acute lung injury(ALI)and acute kidney injury(AKI).Therefore,this article will dis-cuss the effect of m6A modification on the function of immune cells,and its important role in sepsis and its induced multiple or-gan dysfunction diseases by regulating inflammatory signals,py-roptosis,mitochondrial damage and ferroptosis.This will provide new therapeutic targets and strategies for the clinical prevention and treatment of sepsis and its induced multiple organ dysfunc-tion diseases.

Iodine speciations in aquatic environments are affected by dissolved oxygen,redox potential,microbial activity,organic matter decomposition,light reaction,etc.Accurate quantification of iodine speciation can not only help to understand the geochemical cycle of iodine,but also help to trace and study environmental processes.Based on the combination of ion chromatography(IC)and inductively coupled plasma mass spectrometry(ICP-MS),a rapid and sensitive method was established for determining the speciations of iodine in environmental water samples including seawater,river water,lake water,rainwater,groundwater,etc.The results presented here showed that IO3?and I?in seawater were quickly separated and measured within 120 s when using guard column AG22 and 8 mmol/L(NH4)2CO3 as the mobile phase.While for lake water,river water and precipitation samples with high soluble organically bond iodine(SOI),an AS22 separation column(250 mm×4 mm)connected with a guard column and using 50 mmol/L(NH4)2CO3 as mobile phase could effectively separate unknown SOI from IO3? to achieve accurate quantification of IO3?.For accurate correction of iodine measurement signal fluctuations,133Cs was directly added to the(NH4)2CO3 mobile phase as an internal standard.The SOI content was calculated by the total iodine concentrations minus the sum of IO3?and I?.The precision of the established iodine speciation analytical method was better than 3.5%,and the standard addition experiment showed that the analytical method was accurate.When the injection volume was 25 μL,the detection limits were 0.011?0.025 μg/L for IO3? and 0.023?0.031 μg/L for I?,respectively.The method was successfully used to analyze IO3?,SOI and I? in environmental water samples,such as seawater,river water,rainwater and groundwater.

Objective To explore the expression and prognostic value of Smad ubiquitination regulator 2(SMURF2)and protein kinase C receptor 1(RACK1)in epithelial ovarian cancer(EOC)tissues.Methods Cancer tissue specimens of 112 patients with EOC(EOC group)admitted to the hospital from January 2019 to Janu-ary 2021,as well as normal ovarian tissue specimens of 60 patients with other ovarian diseases(control group)who underwent ovarian surgery in the same hospital during the same period were selected.The expressions of SMURF2 and RACK1 in EOC cancer tissues and normal ovarian tissues were detected by real-time fluores-cence quantitative PCR and immunohistochemistry.Pearson correlation analysis was used to analyze the rela-tionship between the expressions of SMURF2 mRNA and RACK1 mRNA in EOC cancer tissues.The Kaplan-Meier survival curve and Log-rank test were used to analyze the differences in survival prognosis among pa-tients with different expressions of SMURF2 and RACK1.Multivariate COX regression analysis was conduc-ted to analyze the influencing factors of survival prognosis in EOC.Results The expression level of SMURF2 mRNA in EOC cancer tissues was lower than that in normal ovarian tissues,and the expression level of RACK1 mRNA was higher than that in normal ovarian tissues,and the differences were statistically signifi-cant(P＜0.05).The expression levels of SMURF2 mRNA and RACK1 mRNA in EOC cancer tissues were negatively correlated(r=-0.764,P＜0.001).The positive rate of SMURF2 in EOC cancer tissues was lower than that in normal ovarian tissues,and the positive rate of RACK1 was higher than that in normal ovarian tis-sues.The difference was statistically significant(P＜0.001).Compared with the cancer tissues of EOC pa-tients with International Federation of Obstetrics and Gynecology(FIGO)stage Ⅰ-Ⅱ and no lymph node metastasis,the positive rate of SMURF2 was lower and the positive rate of RACK1 was higher in the cancer tissues of EOC patients with FIGO stage Ⅲ and lymph node metastasis,and the differences were statistically significant(P＜0.05).The 3-year overall survival rate of EOC patients in the SMURF2 positive group was higher than that in the negative group,and the 3-year overall survival rate of EOC patients in the RACK1 pos-itive group was lower than that in the negative group,and the differences were statistically significant(Log-Rank x2=4.938,5.251;P=0.028,0.018).FIGO stage Ⅲ,lymph node metastasis and positive RACK1 were risk factors affecting the survival prognosis of EOC(P＜0.001),while positive SMURF2 was a protective fac-tor(P＜0.001).Conclusion The expression level of SMURF2 is decreased and the expression level of RACK1 is increased in EOC cancer tissues.Both are related to FIGO stage and lymph node metastasis,and are markers for evaluating the survival prognosis of EOC patients.

Objective To evaluate the clinical outcomes of 514 cases of anterior cruciate ligament(ACL)reconstruction using hamstring tendon autograft and to observe postoperative changes and recovery of the grafts through second-look arthroscopy.Methods This retrospective study collected data from 514 patients who underwent ACL reconstruction with hamstring tendon autograft between May 2015 and June 2018,with a follow-up of at least one year.Knee function recovery and stability were assessed using the Lysholm score,International Knee Documentation Committee(IKDC)score,and Tegner score,along with the pivot shift test and Lachman test.During the second-look arthroscopy,key observations included the synovial coverage,continuity of the reconstructed ligament,and any intra-articular abnormalities.Results The time interval between ACL reconstruction and second-look arthroscopy ranged from 12 to 28 months,with an average of 20 months.Postoperative infection occurred in 2 cases,both of which were successfully treated with arthroscopic debridement and drainage.No other patients experienced infections,graft resorption,or other complications.At the second-look arthroscopy,the Lysholm score significantly improved from(43.56±9.89)preoperative to(92.21±6.12)postoperatively,the difference was statistically significant(P<0.05);The IKDC score increased from(20.32±7.87)to(85.67±10.43),the difference was statistically significant(P<0.05);The Tegner score improved from(4.31±0.82)to(6.61±1.21),the difference was statistically significant(P<0.05).Second-look arthroscopy revealed that the ligament remained intact in 375 patients,with partial tears in 139 patients,ligament tension was maintained in 447 patients,while 67 patients had laxity,the reconstructed ACL graft was deemed to be in good condition in 435 patients and suboptimal in 79 patients,there were significant differences observed in pre-and post-pivot shift test and Lachman test(P<0.05);Among the 514 patients,188(36.58%)successfully returned to sport.Conclusion ACL reconstruction using hamstring tendon autograft effectively restores knee function and stability.In patients followed for more than one years,the grafts show good vascularization and synovial coverage.Emphasis should be placed on systematic postoperative rehabilitation to optimize recovery.

Qing court records show that Arecae Semen was extensively applied. The royal medical records of the Qing Dynasty document nine types of Arecae Semen, with the Palace Museum preserving seven kinds, totaling twelve cultural relics. Historical documents and physical artifacts corroborate each other, providing evidence for the study of the supply channels and court processing of Arecae Semen in the Qing court. According to relevant Qing court archival records, the sources of Arecae Semen used in the imperial court were diverse, including tributes from foreign countries such as Vietnam and Gurkha, annual tributes from local governments in Guangdong, gifts from close aides, and commodities purchased by the Imperial Household Department from civilian shops. The imperial physicians of the Qing court placed great emphasis on the specifications of Arecae Semen slices and were extremely meticulous about their processing. The variety of Arecae Semen slices used in the Qing palace exceeded those recorded in the botanical texts of the era. Compared with the commonly used processing methods for Arecae Semen in the Qing Dynasty, the imperial physicians adjusted the properties and efficacy of the herbs through different processing techniques, based on the patient's condition, constitution, and other factors, in order to meet the clinical treatment needs of the court. The slicing of Arecae Semen in the Qing court required strict control of thickness, with an average thickness of 0.44 mm, which is significantly thinner than the Arecae Semen slices found in today's markets. The texture was softer, making them easier to chew and absorb. Both the Qing court Arecae Semen slices and the Muxiang Binglang Pills focused on the use of authentic medicinal materials, ensuring the quality of the medicine and enhancing the efficacy of Arecae Semen through meticulous selection and preparation.
China ; Drugs, Chinese Herbal/history* ; Humans ; Medicine, Chinese Traditional/history* ; History, 19th Century ; History, Ancient ; History, 17th Century ; History, 18th Century