ObjectiveTo investigate the mechanism by which Luoshi Neiyi prescription treats endometriosis （EMs） through the Toll-like receptor 4 （TLR4）/nuclear factor-κB （NF-κB） signaling pathway. MethodsAnimal experiments were conducted with 50 female SD rats， which were randomized into a sham operation group （10 rats） and a modeling group （40 rats）. An autologous endometrial transplantation method was used for the modeling of EMs. The 36 successfully modeled rats were randomly allocated into four groups （n=9 each）： The model group， the low-dose （7.87 g·kg^-1） Luoshi Neiyi prescription group， the high-dose （15.74 g·kg^-1） Luoshi Neiyi prescription group， and the dienogest （0.20 mg·kg^-1） group. The physiological status and ectopic lesion volumes of rats in each group were observed. Hematoxylin-eosin （HE） staining was used to observe the morphology of the eutopic endometrial tissue. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of inflammatory factors including interleukin-1β （IL-1β） and prostaglandin E₂ （PGE₂） in the serum of EMs rats. Immunohistochemistry was used to detect the expression of matrix metalloproteinase-9 （MMP-9） and vascular endothelial growth factor A （VEGFA） in the eutopic endometrial tissue. Western blot was adopted to determine the protein levels of TLR4， myeloid differentiation factor 88 （MyD88）， phosphorylated nuclear factor-κB （p-NF-κB）/NF-κB， MMP-9， and VEGFA in the eutopic endometrial tissue. In the cell experiments， the cell-counting kit-8 （CCK-8） assay was employed to screen the optimal concentration of Luoshi Neiyi prescription-containing serum， and a scratch assay was performed to assess the migration ability of iheESCs cells. Interventions with Luoshi Neiyi prescription-containing serum， resatorvid （TAK-242， a TLR4 inhibitor）， and lipopolysaccharides （LPS， a TLR4 agonist） were conducted， and Western blot was used to detect the expression of proteins related to the TLR4/NF-κB signaling pathway. ResultsThis experiment successfully replicated 36 EMs rat models. Compared with the sham operation group， the model group exhibited visible ectopic lesions on the abdominal wall and an increase in the writhing response score （P<0.01）. Furthermore， HE staining revealed the model group exhibited a thickened eutopic epithelium， stromal cell disarrangement， and evident infiltration of inflammatory cells. In addition， the model group showed elevated serum levels of IL-1β and PGE₂ （P<0.05， P<0.01）， increased positive expression of MMP-9 and VEGFA in the ectopic endometrial tissue （P<0.01）， and up-regulated protein levels of TLR4， MyD88， p-NF-κB/NF-κB， MMP-9， and VEGFA in the ectopic endometrial tissue （P<0.05， P<0.01）. Compared with the model group， all treatment groups exhibited a reduction in the ectopic lesion volume （P<0.01）. Furthermore， the writhing response scores were decreased in the high-dose Luoshi Neiyi prescription group and the dienogest group （P<0.05， P<0.01）. The pathological state of the ectopic endometrial tissue was alleviated to varying degrees in the treatment groups. Low-dose Luoshi Neiyi prescription reduced serum PGE₂ levels， and high-dose Luoshi Neiyi prescription and dienogest decreased serum IL-1β and PGE₂ levels in EMs rats （P<0.05， P<0.01）. The treatment groups showed decreased positive expression of MMP-9 and VEGFA in the ectopic endometrial tissue （P<0.05， P<0.01） and down-regulated protein levels of TLR4， MyD88， p-NF-κB/NF-κB， MMP-9， and VEGFA in the ectopic endometrial tissue （P<0.05， P<0.01）. The cell experiments showed that 5%， 10%， and 20% Luoshi Neiyi prescription-containing sera significantly reduced the viability and inhibited the migration of iheESCs. Compared with the control group， 5%， 10%， and 20% Luoshi Neiyi prescription-containing serum groups and the TAK-242 group showed reduced protein levels of TLR4， MyD88， p-NF-κB/NF-κB， MMP-9， and VEGFA （P<0.01）. Compared with the control group， the LPS group showed increased expression of the above proteins （P<0.01）. Compared with the LPS group， the LPS+5%， 10%， and 20% Luoshi Neiyi prescription-containing serum groups showed reduced expression of the above proteins （P<0.05， P<0.01）. ConclusionLuoshi Neiyi prescription may modulate the TLR4/NF-κB signaling pathway to reduce the inflammatory response and histopathological damage in the eutopic endometrium and suppress the adhesive， invasive， and angiogenic biological processes in the ectopic endometrial tissue， thereby exerting its therapeutic effect on EMs.

Objective:To investigate the therapeutic effect of artesunate on mice with acute Toxoplasma gondii ( T. gondii) infection. Methods:Based on body weight (16 - 18 g), sixty-four C57BL/6 female mice aged 6 - 8 weeks were divided into 4 groups by random number table method: uninfected control group without treatment; T. gondii-infected group (Tg group), each mouse was intraperitoneally (i.p.) infected with 100 tachyzoites of T. gondii RH strain; T. gondii-infected + artesunate treatment group (Tg + ART group), 3 hours after each mouse i.p. infected with 100 tachyzoites of T. gondii RH strain, artesunate solution was i.p. injected at a dose of 30 mg/kg, once a day for a total of 7 consecutive days; T. gondii-infected + sulfadiazine treatment group (Tg + SDZ group), 3 hours after each mouse i.p. infected with 100 tachyzoites of T. gondii RH strain, sulfadiazine solution was orally administrated at a dose of 100 mg/kg, once a day for a total of 7 consecutive days. There were 16 mice in each group, in which 10 mice were used to observe survival time and 6 mice were used to monitor body weight and collect tissue samples. Mice were weighed every day from day 1 post infection (p.i.); mice were sacrificed at day 7 p.i., the liver weights of mice were weighed and the liver indexes were calculated; liver tissues were paraffin-embedded, sectioned, and stained with hematoxylin-eosin (HE), and the pathological changes of liver tissues of mice in each group were observed under a light microscope. The expression levels of T. gondii major surface antigen 1 (SAG1) in the liver tissues of mice in each group were detected by real-time quantitative PCR for evaluating parasite load. Results:All mice in the uninfected control group were survived. The survival time was 7 - 9 days in Tg group, 8 - 11 days in Tg + ART group, and 9 - 13 days in Tg + SDZ group. Compared with Tg group, the survival times of mice in Tg + ART group and Tg + SDZ group were significantly longer ( P < 0.05). On day 7 p.i., compared with uninfected control group, Tg + ART group or Tg + SDZ group, the body weight of mice in Tg group was lower ( P < 0.05); however, there was no significant difference of body weight in Tg + ART group and Tg + SDZ group compared with uninfected control group ( P > 0.05). Compared with Tg group, Tg + ART group and Tg + SDZ group had lower liver indexes and SAG1 mRNA expression levels in the liver tissues ( P < 0.05 or < 0.001), and liver histopathological changes were milder. Compared with Tg + SDZ group, there was no significant difference in both liver index and SAG1 mRNA expression level in the liver tissue of Tg + ART group ( P > 0.05). Conclusion:Artesunate solution i.p. injection can prolong the survival time, reduce parasite load in the liver, and attenuate hepatic pathological damage, to a certain extent, of mice with acute T. gondii infection.

Objective:To investigate the regulation and mechanism of chronic Trichinella spiralis ( Ts) infection on liver immunopathology in mice co-infected with Plasmodium berghei ANKA( PbA). Methods:According to body weight, 64 specific pathogen free female Kunming mice (6 - 8 weeks old, weighting 22 - 25 g) were divided into 4 groups by using random number table method. Control group: uninfected; Ts group: mice were mono-infected with 30 Ts larvae by oral gavage on day 0; PbA group: mice were mono-infected with 1 × 10 6PbA-infected red blood cells in 0.1 ml of phosphate buffer (PBS) administered by intraperitoneal injection on day 121; co-infected ( Ts+PbA) group: mice were infected with 30 Ts larvae by oral gavage and intraperitoneal injected with 1 × 10 6PbA-infected red blood cells in 0.1 ml PBS on day 121 after Ts infection. There were 16 mice in each group, in which 10 mice in each group were monitored for the survival rate. The peripheral red blood cell parasitemia of PbA group and Ts + PbA group were monitored every other day by light microscope examination of Giemsa-stained thin tail-blood smears from day 3 after PbA infection. Mice were sacrificed at day 135 after Ts infection and/or at day 15 after PbA infection, the mouse body weight and liver weight were measured, and the liver index were calculated. Ts-infected mice were monitored by a light microscope examination of diaphragm compression slide. Under a light microscope, the liver pathology and liver fibrosis of mice were observed and compared with hematoxylin-eosin (HE) staining and Sirius red staining, respectively. The F4/80 + Kupffer cells in liver of mice were examined by immunohistochemical staining. Results:After infection with Ts or PbA, Ts larvae cysts were observed in diaphragm tissues and PbA were observed in red blood cells under the light microscope. After PbA infection, there was no significant difference in survival rate between PbA group and Ts+ PbA group ( P > 0.05). Compared with PbA group, the peripheral red blood cell parasitemia was significantly decreased in Ts+ PbA group on days 11 and 15 after PbA infection (%: 27.104 ± 7.623 vs 45.032 ± 9.849, 60.218 ± 2.776 vs 76.778 ± 6.351, P < 0.05), and the liver index and the liver pathology score were significantly decreased in Ts+ PbA group ( P < 0.05). Sirius red staining showed that the positive area of liver fibrosis in Ts+ PbA group was significantly higher than that in PbA group ( P < 0.05). Immunohistochemical staining showed that the average optical density value of F4/80 + Kupffer cells in Ts+ PbA group was significantly higher than that in PbA group ( P < 0.01). Conclusion:Chronic Ts infection may reduce the peripheral red blood cell parasitemia, increase F4/80 + Kupffer cells expression in liver, and attenuate liver pathology in mice co-infected with PbA.

OBJECTIVE:To establish fingerprint of Zhizi jinhua pills(ZZJHW)and analyze the relationship of it with in vitro antioxidant activity,in order to provide the basis for the quality control of them. METHODS:HPLC method was adopted. The sep-aration was performed on a Sinochrom ODS-BP C18(200 mm×4.6 mm,5 μm)column with mobile phase consisted of 0.2% acetic acid(containing 3 mmol/L sodium heptanesulfonate solution)-acetonitrile(gradient elution)at the detection wavelength of 254 nm and flow rate of 0.8 ml/min. The column temperature was controlled at 38 ℃,and injection volume was 10 μl. The“Chromato-graphic Fingerprint Similarity Evaluation System for TCM”(2012.130723 edition) issued by Chinese Pharmacopoeia Commission was used to evaluate the similarity of the 12 batches of ZZJHW using baicalin as reference peak so as to attribute the common peak of fingerprint. DPPH free radical scavenging assay was used to investigate the in vitro antioxidant activity of 12 batches of ZZJHW,and the relationship between its fingerprint and antioxidant activity was studied. RESULTS:The fingerprint of 12 batches of ZZJHW was established and the similarity between the fingerprint of ZZJHW with their reference fingerprint were all above 0.9 (except S1,S2,S3,S12). 30 common peaks were marked,all of which were assigned to the herbs. Antioxidant experiment result showed the differences in the antioxidant capacity among different batches of ZZJHW;spectrum effect relationship showed that 13 common peaks were positively related with oxidation activity and 17 common peaks negatively related with it;among known com-ponents,oxidation activity components were mainly from Lonicera japonica,Scutellaria baicalensis and Rheum palmatum. CON-CLUSIONS:The spectrum effect relationship of established fingerprint with its antioxidant activity can provide reference for the quality control of ZZJHW.