The most common medications for the treatment of zoonotic toxoplasmosis are pyrimethamine and sulfadiazine, which may cause serious undesirable side effects. Thus, there is an urgent need to develop novel therapeutics. Baicalein (BAI, C₁₅H₁₀O₅) has been shown to perform well against protozoan parasites including Leishmania and Cryptosporidium. In this study, the inhibition efficacy of BAI on Toxoplasma gondii was evaluated using plaque, invasion, and intracellular proliferation assays. BAI effectively inhibited T. gondii (half-maximum inhibitory concentration (IC₅₀)=6.457×10^-5 mol/L), with a reduced invasion rate (33.56%) and intracellular proliferation, and exhibited low cytotoxicity (half-maximum toxicity concentration (TC₅₀)=5.929×10^-4 mol/L). Further investigation using a mouse model shed light on the inhibitory efficacy of BAI against T. gondii, as well as the potential mechanisms underlying its anti-parasitic effects. The survival time of T. gondii-infected ICR mice treated with BAI was remarkably extended, and their parasite burdens in the liver and spleen were greatly reduced compared with those of the negative control group. Histopathological examination of live sections revealed effective therapeutic outcomes in the treatment groups, with no notable pathological alterations observed. Furthermore, alterations in cytokine levels indicated that BAI not only effectively suppressed the growth of T. gondii but also prevented excessive inflammation in mice. Collectively, these findings underscore the significant inhibitory efficacy of BAI against T. gondii, positioning it as a promising alternative therapeutic agent for toxoplasmosis.
Animals ; Toxoplasma/drug effects* ; Flavanones/therapeutic use* ; Mice ; Antiparasitic Agents/therapeutic use* ; Mice, Inbred ICR ; Toxoplasmosis/drug therapy* ; Female

Objective:A camelid natural nanobody library was screened to target S100 calcium-binding protein B (S100B) for obtaining high-affinity and specific nanobodies (Nbs), which provided a molecular basis for the early diagnosis and prognostic treatment of malignant melanoma.Methods:In this study, affinity panning was employed to isolate S100B-specific nanobodies with unique complementarity-determining region 3 (CDR3) sequences from a camelid natural nanobody library. The selected Nbs were expressed in a prokaryotic system and purified via Ni-NTA affinity chromatography. The affinity, specificity, and diagnostic potential of the Nbs were evaluated using enzyme-linked immunosorbent assay (ELISA), western blot, and bio-layer interferometry (BLI).Results:A camelid natural anti-S100B nanobody library with a capacity of 1.91×10 8 CFU was successfully constructed. Affinity panning yielded 30 S100B-specific Nbs, among which Nb107, Nb122, Nb212, and Nb324 with distinct CDR3 sequences were selected for expression. Following Ni-NTA purification, all four anti-S100B Nbs exhibited high purity. Western blot analysis confirmed their ability to recognize recombinant S100B. ELISA and BLI analyses revealed that Nb212 demonstrated high affinity (1.96×10 -11 mol). Additionally, Nb107, Nb122, and Nb212 exhibited broad-spectrum recognition capabilities, binding to various tumor cell lines (Hepa1-6, GL261, 4T1, CT26) as well as murine/human melanoma cells. These Nbs also effectively bound to native murine/human antigens in serum samples from melanoma (A375, B16F10) mouse models. Conclusions:Specific anti-S100B Nbs are successfully screened and expressed, demonstrating not only recognition of native conformational antigens but also broad-spectrum binding and high affinity. These findings highlight their significant potential for developing early diagnostic assays and broad-spectrum targeted vaccines or therapeutics against diverse tumor cells.

Objective:A camelid natural nanobody library was screened to target S100 calcium-binding protein B (S100B) for obtaining high-affinity and specific nanobodies (Nbs), which provided a molecular basis for the early diagnosis and prognostic treatment of malignant melanoma.Methods:In this study, affinity panning was employed to isolate S100B-specific nanobodies with unique complementarity-determining region 3 (CDR3) sequences from a camelid natural nanobody library. The selected Nbs were expressed in a prokaryotic system and purified via Ni-NTA affinity chromatography. The affinity, specificity, and diagnostic potential of the Nbs were evaluated using enzyme-linked immunosorbent assay (ELISA), western blot, and bio-layer interferometry (BLI).Results:A camelid natural anti-S100B nanobody library with a capacity of 1.91×10 8 CFU was successfully constructed. Affinity panning yielded 30 S100B-specific Nbs, among which Nb107, Nb122, Nb212, and Nb324 with distinct CDR3 sequences were selected for expression. Following Ni-NTA purification, all four anti-S100B Nbs exhibited high purity. Western blot analysis confirmed their ability to recognize recombinant S100B. ELISA and BLI analyses revealed that Nb212 demonstrated high affinity (1.96×10 -11 mol). Additionally, Nb107, Nb122, and Nb212 exhibited broad-spectrum recognition capabilities, binding to various tumor cell lines (Hepa1-6, GL261, 4T1, CT26) as well as murine/human melanoma cells. These Nbs also effectively bound to native murine/human antigens in serum samples from melanoma (A375, B16F10) mouse models. Conclusions:Specific anti-S100B Nbs are successfully screened and expressed, demonstrating not only recognition of native conformational antigens but also broad-spectrum binding and high affinity. These findings highlight their significant potential for developing early diagnostic assays and broad-spectrum targeted vaccines or therapeutics against diverse tumor cells.

Objective:To explore the clinical efficacy of immunoadsorption in highly sensitized kidney transplant (KT) candidates.Methods:From September 2019 to April 2023, the relevant clinical data were retrospectively reviewed for 26 highly sensitized KT recipients. Protein A immunoadsorption desensitization therapy was offered after KT. The effect of immunosorbent on reducing anti-human leukocyte antigen (HLA) antibodies was summarized. And operative success rate and postoperative complication incidence were calculated.Results:The mean number of treatment session was (10.76±5.53). The highest level of HLA-Ⅰ antibody mean fluorescence intensity (MFI) dropped from (17 921±4 442) to (7 333±6 434) with a decline of 59% and HLA-Ⅱ antibody MFI decreased from (21 135±5 245) to (10 989±7 627) with a decline of 48%. The differences were statistically significant (both P<0.001). All kidneys were harvested from cadavers. The complications were acute antibody mediated rejection (7 cases), perioperative pulmonary infection (3 cases) and myelosuppression (2 cases). The average follow-up period was (30.8±12.6) month. The graft survival rate was 88.5% (23/26) and the recipient survival rate 100% (26/26) . Conclusions:Immunoadsorption therapy can effectively reduce HLA antibody in highly sensitized KT candidates, thereby increasing the probability of successful KT. In terms of safety, immunosorbent therapy may boost the potential risks of infection and myelosuppression. It requires heightened attention.

Objective:To investigate the correlation between the expression level of serum exosome miR-29C and postoperative cognitive dysfunction (POCD) in elderly patients undergoing surgery.Methods:A total of 119 elderly patients who underwent elective spinal surgery in the Second Hospital of Shanxi Medical University from January 2021 to January 2022 were selected and scored on the Montreal Cognitive Assessment (MoCA) Scale 1 day before surgery and 1, 7 and 21 days after surgery. The selected patients were divided into POCD group (51 cases) and non-POCD group (68 cases) according to whether the MoCA Scale score decreased ≥2 points 1 day before surgery and 1 day after surgery. S100-β, neuron-specific enolase (NSE) levels and serum exosome miR-29C expression levels were detected and analyzed in all patients 1 day before and 1 day after surgery. Pearson correlation analysis showed the correlation between MoCA Scale score and S100-β, NSE and miR-29C. Receiver operating characteristic (ROC) curve was used to evaluate the predictive value of S100-β, NSE and miR-29C for POCD occurrence in elderly patients undergoing surgery.Results:The score of MoCA Scale in POCD group were significantly decreased 1, 7 and 21 days after surgery compared with 1 day before surgery (all P<0.05), while the score of MoCA Scale in non-POCD group were significantly decreased only 1 day after surgery compared with 1 day before surgery ( P<0.05). The levels of S100-β and NSE and the expression level of serum exosome miR-29C in 2 groups were significantly increased 1 day after surgery compared with 1 day before surgery (all P<0.05). Moreover, the levels of S100-β and NSE and the expression level of serum exosome miR-29C in POCD group were significantly higher than those in non-POCD group 1 day after surgery (all P<0.05). There was a negative correlation between the MoCA Scale score and the expression level of serum exosome miR-29C 1 day after surgery in the POCD group ( P<0.05). ROC curve analysis showed that the expression levels of NSE, S100-β and exosome miR-29C 1 day after surgery predicted the risk of POCD in elderly surgical patients with area under the curve (AUC) of 0.891, 0.908 and 0.918, respectively. Conclusions:The occurrence of POCD in elderly patients with surgery is related to the increase of the expression level of serum exosome miR-29C, and the expression level of serum exosome miR-29C is negatively correlated with MoCA Scale score. Early monitoring of the miR-29C expression level can provide a basis for the occurrence and development of postoperative POCD in elderly patients, disease diagnosis and clinical intervention.

Toxoplasma gondii is a worldwide parasite that can infect almost all kinds of mammals and cause fatal toxoplasmosis in immunocompromised patients. Apoptosis is one of the principal strategies of host cells to clear pathogens and maintain organismal homeostasis, but the mechanism of cell apoptosis induced by T. gondii remains obscure. To explore the apoptosis influenced by T. gondii, Vero cells infected or uninfected with the parasite were subjected to apoptosis detection and subsequent dual RNA sequencing (RNA-seq). Using high-throughput Illumina sequencing and bioinformatics analysis, we found that pro-apoptosis genes such as DNA damage-inducible transcript 3 (DDIT3), growth arrest and DNA damage-inducible α (GADD45A), caspase-3 (CASP3), and high-temperature requirement protease A2 (HtrA2) were upregulated, and anti-apoptosis genes such as poly(adenosine diphosphate (ADP)-ribose) polymerase family member 3 (PARP3), B-cell lymphoma 2 (Bcl-2), and baculoviral inhibitor of apoptosis protein (IAP) repeat containing 5 (BIRC5) were downregulated. Besides, tumor necrosis factor (TNF) receptor-associated factor 1 (TRAF1), TRAF2, TNF receptor superfamily member 10b (TNFRSF10b), disabled homolog 2 (DAB2)-interacting protein (DAB2IP), and inositol 1,4,5-trisphosphate receptor type 3 (ITPR3) were enriched in the upstream of TNF, TNF-related apoptosis-inducing ligand (TRAIL), and endoplasmic reticulum (ER) stress pathways, and TRAIL-receptor 2 (TRAIL-R2) was regarded as an important membrane receptor influenced by T. gondii that had not been previously considered. In conclusion, the T. gondii RH strain could promote and mediate apoptosis through multiple pathways mentioned above in Vero cells. Our findings improve the understanding of the T. gondii infection process through providing new insights into the related cellular apoptosis mechanisms.

Toxoplasma gondii is a worldwide parasite that can infect almost all kinds of mammals and cause fatal toxoplasmosis in immunocompromised patients. Apoptosis is one of the principal strategies of host cells to clear pathogens and maintain organismal homeostasis, but the mechanism of cell apoptosis induced by T. gondii remains obscure. To explore the apoptosis influenced by T. gondii, Vero cells infected or uninfected with the parasite were subjected to apoptosis detection and subsequent dual RNA sequencing (RNA-seq). Using high-throughput Illumina sequencing and bioinformatics analysis, we found that pro-apoptosis genes such as DNA damage-inducible transcript 3 (DDIT3), growth arrest and DNA damage-inducible α (GADD45A), caspase-3 (CASP3), and high-temperature requirement protease A2 (HtrA2) were upregulated, and anti-apoptosis genes such as poly(adenosine diphosphate (ADP)-ribose) polymerase family member 3 (PARP3), B-cell lymphoma 2 (Bcl-2), and baculoviral inhibitor of apoptosis protein (IAP) repeat containing 5 (BIRC5) were downregulated. Besides, tumor necrosis factor (TNF) receptor-associated factor 1 (TRAF1), TRAF2, TNF receptor superfamily member 10b (TNFRSF10b), disabled homolog 2 (DAB2)‍-interacting protein (DAB2IP), and inositol 1,4,5-trisphosphate receptor type 3 (ITPR3) were enriched in the upstream of TNF, TNF-related apoptosis-inducing ligand (TRAIL), and endoplasmic reticulum (ER) stress pathways, and TRAIL-receptor 2 (TRAIL-R2) was regarded as an important membrane receptor influenced by T. gondii that had not been previously considered. In conclusion, the T. gondii RH strain could promote and mediate apoptosis through multiple pathways mentioned above in Vero cells. Our findings improve the understanding of the T. gondii infection process through providing new insights into the related cellular apoptosis mechanisms.
Animals ; Apoptosis ; Chlorocebus aethiops ; Gene Expression Profiling ; Humans ; Mammals/genetics* ; Toxoplasma/genetics* ; Toxoplasmosis/pathology* ; Vero Cells ; ras GTPase-Activating Proteins/genetics*

In recent years, the regulatory role of microRNAs in liver pathological process has attracted more and more attention. In viral hepatitis and steatohepatitis, microRNA-335 (miRNA-335) regulates the progression of hepatitis via the transcription factor sex-determining region Y-box 4; in the development and progression of progressive liver fibrosis and liver cancer, miRNA-335 affects collagen production, deposition, and degradation in the liver via the target genes including hypoxia-inducible factor 1α, phosphatase and tensin homologue, Rho-associated coiled-coil containing protein kinase 1, plasminogen activator inhibitor-1, twist family bHLH transcription factor 1, and mesenchymal-epithelial transition factor and thus regulates the migration and invasion of hepatic stellate cells and hepatoma cells. This article summarizes the research advances in the role of miRNA-335 in hepatitis, liver fibrosis, and liver cancer in recent years, and based on existing data, it is pointed out that the miRNA-335/QSOX1 regulatory axis may mediate artesunate against schistosomal liver fibrosis by inhibiting hepatic stellate cell activation, so as to provide new ideas for the treatment of liver fibrosis and other liver diseases.

Objective:To investigate the risk factors of deep venous thrombosis in patients before hip arthroplasty, and to explore the possible causes of postoperative thrombosis.Methods:The clinical data of 361 patients with hip arthroplasty treated in the Department of Orthopaedic Trauma of Xi′an Honghui Hospital from September 2015 to December 2019 were studied retrospectively, including 102 males and 259 females, aged 65 to 94 years, and the average age was 72.25 years old. All fracture patients were given subcutaneous injection of low molecular weight heparin calcium to prevent lower extremity thrombosis. The deep veins of both lower extremities were examined before and after operation. The general data of the two groups of patients were collected and recorded, including age, sex, whether complicated with medical diseases (essential hypertension, type 2 diabetes, coronary heart disease), serological indexes, time from injury to admission, and time from admission to operation. The software of SPSS 19.0 was used for statistical analysis.Results:The incidence of lower limb DVT, before operation was 29.92%, including 26 males (24.07%) and 82 females (75.93%). The results of multivariate logistic regression analysis showed that diabetes mellitus ( OR=2.127, 95% CI: 1.134-3.989, P=0.019), coronary heart disease ( OR=1.692, 95% CI: 1.056-2.713, P=0.029) and the time from injury to admission ( OR=1.677, 95% CI: 1.037-2.712, P=0.035) were independent risk factors for DVT in elderly patients undergoing hip arthroplasty. The incidence of lower limb DVT, after operation was 46.54%. After operation, proximal thrombus were occurred in 2 cases (1.19%), distal thrombus in 143 cases (85.12%), and mixed thrombus in 23 cases (13.69%). Postoperative thrombus was ipsilateral to the fracture limb in 84 cases (50.00%), thrombus was located in the healthy side of the fracture in 19 cases (11.31%), and DVT occurred in 65 cases (38.69%) in both lower limbs. Conclusions:Delayed admission longed than 48 hours, coronary heart disease and diabetes mellitus are the risk factors for the formation of DVT. The thrombus that existed before operation and did not disappear after operation accounted for 48.81% of the total incidence of postoperative thrombosis, and the new thrombus accounted for 51.19% of the total incidence of postoperative thrombosis. For the elderly patients with femoral neck fracture undergoing hip arthroplasty, ultrasonic examination of both lower limbs should be performed before and after operation to find the changes of thrombus in time and do a good job of prevention and treatment.

Objective To investigate the effect of exercise prescription on foot perfusion and ulcer healing in patients with diabetic foot ulcer (DFU) accompanied with peripheral arterial disease (PAD).Methods Sixty patients with DFU and PAD were divided into exercise group and control group.The patients in the exercise group took exercise therapy training,ensuring that they could follow the exercise prescription.The patients in the control group had no exercise requirements.After the treatment for 12 weeks,ankle brachial index (ABI),transcutaneous oxygen tension (TcPO2),skin temperature,body mass index (BMI),HbA1C,and the ulcer healing rate in the two groups were compared,the satisfaction and compliance in the exercise group were evaluated,and the adverse events of the treatment were recorded.Results After the treatment for 12 weeks,the ABI,TcPO2,and foot skin temperature in the two groups increased and HbA1C decreased(P＜0.05 or P＜0.01).TcPO2 increased 5.25 mm Hg(1 mm Hg=0.133kPa),skin temperature increased 0.45℃,and BMI decreased 0.69 kg/m2 in the exercise group,while TcPO2 increased 2.59 mm Hg,skin temperature increased 0.28℃,and BMI increased 0.02 kg/m2 in the control group,showing significant differences in the three index changes between two groups (P＜0.01).The healing rate in the exercise group was higher than that in the control group (53.6％ vs 25.9％,P ＜0.05).The patients'compliance and satisfaction to the exercise therapy were 90％ and 94％,respectively.The incidence of adverse events in the exercise group had no causal relationship with the exercise prescription.Conclusion The exercise prescription in the study can improve the foot perfusion,and promote ulcer healing.It has the advantage of high safety,patients'compliance and satisfaction.