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.

Objective To investigate the changes in eosinophil counts and the activation of microglial cells in the brain tissues of mice at different stages of Angiostrongylus cantonensis infection, and to examine the role of microglia in regulating the progression of angiostrongyliasis and unravel the possible molecular mechanisms. Methods Fifty BALB/c mice were randomly divided into the control group and the 7-d, 14-d, 21-day and 25-d infection groups, of 10 mice in each group. All mice in infection groups were infected with 30 stage III A. cantonensis larvae by gavage, and animals in the control group was given an equal amount of physiological saline. Five mice were collected from each of infection groups on days 7, 14, 21 d and 25 d post-infection, and 5 mice were collected from the control group on the day of oral gavage. The general and focal functional impairment was scored using the Clark scoring method to assess the degree of mouse neurological impairment. Five mice from each of infection groups were sacrificed on days 7, 14, 21 d and 25 d post-infection, and 5 mice from the control group were sacrificed on the day of oral gavage. Mouse brain tissues were sampled, and the pathological changes of brain tissues were dynamically observed using hematoxylin and eosin (HE) staining. Immunofluorescence staining with eosinophilic cationic protein (ECP) and ionized calcium binding adaptor molecule 1 (Iba1) was used to assess the degree of eosinophil infiltration and the counts of microglial cells in mouse brain tissues in each group, and the morphological parameters of microglial cells (skeleton analysis and fractal analysis) were quantified by using Image J software to determine the morphological changes of microglial cells. In addition, the expression of M1 microglia markers Fcγ receptor III (Fcgr3), Fcγ receptor IIb (Fcgr2b) and CD86 antigen (Cd86), M2 microglia markers Arginase 1 (Arg1), macrophage mannose receptor C-type 1 (Mrc1), chitinase-like 3 (Chil3), and phagocytosis genes myeloid cell triggering receptor expressed on myeloid cells 2 (Trem2), CD68 antigen (Cd68), and apolipoprotein E (Apoe) was quantified using real-time quantitative reverse transcription PCR (RT-qPCR) assay in the mouse cerebral cortex of mice post-infection. Results A large number of A. cantonensis larvae were seen on the mouse meninges surface post-infection, and many neuronal nuclei were crumpled and deeply stained, with a large number of bleeding points in the meninges. The median Clark scores of mouse general functional impairment were 0 (interquartile range, 0), 0 (interquartile range, 0.5), 6 (interquartile range, 1.0), 14 (interquartile range, 8.5) points and 20 (interquartile range, 9.0) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.45, P < 0.01), and the median Clark scores of mouse focal functional impairment were 0 (interquartile range, 0), 2 (interquartile range, 2.5), 7 (interquartile range, 3.0), 18 (interquartile range, 5.0) points and 25 (interquartile range, 6.5) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.72, P < 0.01). The mean scores of mice general and focal functional impairment were all higher in the infection groups than in the control group (all P values < 0.05). Immunofluorescence staining showed a significant difference in the eosinophil counts in mouse brain tissues among the five groups (F = 40.05, P < 0.000 1), and the eosinophil counts were significantly higher in mouse brain tissues in the 14-d (3.08 ± 0.78) and 21-d infection groups (5.97 ± 1.37) than in the control group (1.00 ± 0.28) (both P values < 0.05). Semi-quantitative analysis of microglia immunofluorescence showed a significant difference in the counts of microglial cells among the five groups (F = 17.66, P < 0.000 1), and higher Iba1 levels were detected in mouse brain tissues in 14-d (5.75 ± 1.28), 21-d (6.23 ± 1.89) and 25-d infection groups (3.70 ± 1.30) than in the control group (1.00 ± 0.30) (all P values < 0.05). Skeleton and fractal analyses showed that the branch length [(162.04 ± 34.10) μm vs. (395.37 ± 64.11) μm; t = 5.566, P < 0.05] and fractal dimension of microglial cells (1.30 ± 0.01 vs. 1.41 ± 0.03; t = 5.266, P < 0.05) were reduced in mouse brain tissues in the 21-d infection group relative to the control group. In addition, there were significant differences among the 5 groups in terms of M1 and M2 microglia markers Fcgr3 (F = 48.34, P < 0.05), Fcgr2b (F = 55.46, P < 0.05), Cd86 (F = 24.44, P < 0.05), Arg1 (F = 31.18, P < 0.05), Mrc1 (F = 15.42, P < 0.05) and Chil3 (F = 24.41, P < 0.05), as well as phagocytosis markers Trem2 (F = 21.19, P < 0.05), Cd68 (F = 43.95, P < 0.05) and Apoe (F = 7.12, P < 0.05) in mice brain tissues. Conclusions A. cantonensis infections may induce severe pathological injuries in mouse brain tissues that are characterized by massive eosinophil infiltration and persistent activation of microglia cells, thereby resulting in progressive deterioration of neurological functions.

Objective:To study echinococcosis in the population and canine Echinococcus infection in Yushu City, Qinghai Province, and to explore the current epidemic situation and main transmission species of Echinococcus. Methods:In June 2023, a multi-stage sampling method was used to select 2 villages each in Shanglaxiu Township and Longbao Town, Yushu City, Qinghai Province. Each village included at least 100 permanent residents who had lived locally for at least 1 year and were 2 years old or older as the survey subjects. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum antibodies against Echinococcus larvae in the population, and B-mode ultrasound was used for abdominal organ scanning. Meanwhile, on the main roads of Shanglaxiu Township and Longbao Town, canine feces were collected in designated areas at intervals. ELISA was used to detect the antigen of canine fecal Echinococcus, and PCR was used to detect the types of parasites ( Echinococcus multilocularis, Echinococcus granulosus and Echinococcus shiquicus). Results:A total of 511 residents were investigated in Yushu City, and the positive rate of serum Echinococcus larvae antibodies in the population was 26.22% (134/511), and the detection rate of echinococcosis B-mode ultrasound was 1.37% (7/511). Among them, the detection rates of B-mode ultrasound for cystic echinococcosis (CE) and alveolar echinococcosis (AE) were 1.17% (6/511) and 0.20% (1/511), respectively. The positive rate of Echinococcus antigen in 543 canine feces detected by ELISA was 12.89% (70/543). PCR was used to test 497 canine feces, and the detection rate of Echinococcus was 3.02% (15/497). Among them, the detection rate of Echinococcus multilocularis was higher than that of Echinococcus granulosus [2.82% (14/497) vs 0.20% (1/497)], and the difference was statistically significant (χ 2 = 11.44, P < 0.001). No Echinococcus shiquicus was detected. Conclusions:The positive rates of Echinococcus larvae antibodies in the population and canine Echinococcus antigen in Yushu City, Qinghai Province are both relatively high. There is a mixed epidemic of CE and AE, with Echinococcus multilocularis being the main species.

Objective:Comparative analysis of the mNUTRIC and NRS-2002 scores for evaluating nutritional risk and predicting clinical outcomes in end stage liver disease patients.Method:A retrospective cohort study method was used to screen 114 cases with end-stage liver disease admitted to the intensive care unit (ICU) of the First Hospital of Lanzhou University from December 1, 2016 to March 31, 2021 according to the inclusion and exclusion criteria. The patient's demographic data, blood routine, blood biochemical indexes, coagulation function indexes, arterial blood gas analysis and imaging examination data were collected. The mNUTRIC score, NRS-2002 score, sequential organ failure (SOFA) score, model for end-stage liver disease (MELD) score, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, Child-Pugh grade, and clinical outcomes at 28 and 90 days at 24 h post-ICU admission were collected. The differences in clinical indicators between the mNUTRIC high group (≥5 points) and the low group, and the NRS-2002 high group (≥3 points) and the low group were compared. Spearman correlation analysis was used to explore the correlation between the mNUTRIC score and NRS-2002 score, clinical indicators, and 28 and 90-day mortality rates. Multivariate logistic regression analysis was used to determine the risk factors associated with 28-day and 90-day mortality in patients. The value of mNUTRIC score and NRS-2002 score in assessing the clinical outcomes of patients with end-stage liver disease was explored by receiver operating characteristic (ROC) curve.Results:The clinical indicators related to nutritional status of patients were worse in the high-mNUTRIC group than those in the low-mNUTRIC group, and the 28-day and 90-day mortality rates were significantly higher than those in the low-mNUTRIC group [89.0%(65/73) vs. 29.2%(12/41), 97.2%(71/73) vs. 39.0%(16/41), P<0.001]. There was no statistically significant difference in the incidence rate of hepatic encephalopathy, esophageal variceal bleeding, and ascites between the high and low mNUTRIC group. The clinical indicators related to nutritional status were worse in the high-NRS-2002 group than those in the low-NRS-2002 group of patients, and the 28-day and 90-day mortality rates were significantly higher than those in the low-group [73.0%(73/100) vs. 4/14, 81.0%(81/100) vs. 6/14, P=0.008, 0.004]. The NRS-2002 high-score group did not differ significantly from the low-score group in terms of hepatic encephalopathy, esophagogastric variceal bleeding, or ascites prevalence. Patient's age, white blood cell count (WBC), urea nitrogen (BUN), creatinine (UREA), uric acid (UA), total cholesterol (TG), Child-Pugh, MELD, SOFA, APACHE Ⅱscores were significantly positively correlated with the mNUTRIC score. Conversely, albumin (Alb) and Glasgow Coma Scale (GCS) were significantly negatively correlated. Patient's age, WBC, CREA, BUN, UREA, UA, Child-Pugh, MELD, SOFA, APACHE Ⅱwere significantly positively correlated with the NRS-2002 score.Conversely, albumin (Alb) and Glasgow Coma Scale (GCS) were significantly negatively correlated ( P<0.05). The 28-day and 90-day mortality rates of patients increased with the increase in the mNUTRIC scores. The mNUTRIC score was an independent predictor of death within 28 and 90 days in patients with end-stage liver disease. The area under the curve (AUC) of mNUTRIC for predicting patient death at 28 days was 0.864 (95% CI: 0.794-0.934). The AUC of NRS-2002 for predicting patient death at 28 days was 0.683 (95% CI: 0.573-0.792). The AUC of the two indicators combined for predicting patient death at 28 days was 0.868 (95% CI: 0.799-0.936). The AUC of mNUTRIC for predicting patient death at 90 days was 0.915 (95% CI: 0.861-0.969). The AUC of NRS-2002 for predicting patient death at 90 days was 0.715 (95% CI: 0.599-0.832). The AUC of the two indicators combined for predicting patient death at 90 days was 0.922 (95% CI: 0.871-0.972). Conclusion:mNUTRIC score and NRS-2002 score can better evaluate the nutritional status in patients with end-stage liver disease. The mNUTRIC score is a good predictor of 28-day and 90-day mortality in patients with end-stage liver disease, and its application value efficacy is enhanced when combined with NRS-2002.

Objective:To study echinococcosis in the population and canine Echinococcus infection in Yushu City, Qinghai Province, and to explore the current epidemic situation and main transmission species of Echinococcus. Methods:In June 2023, a multi-stage sampling method was used to select 2 villages each in Shanglaxiu Township and Longbao Town, Yushu City, Qinghai Province. Each village included at least 100 permanent residents who had lived locally for at least 1 year and were 2 years old or older as the survey subjects. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum antibodies against Echinococcus larvae in the population, and B-mode ultrasound was used for abdominal organ scanning. Meanwhile, on the main roads of Shanglaxiu Township and Longbao Town, canine feces were collected in designated areas at intervals. ELISA was used to detect the antigen of canine fecal Echinococcus, and PCR was used to detect the types of parasites ( Echinococcus multilocularis, Echinococcus granulosus and Echinococcus shiquicus). Results:A total of 511 residents were investigated in Yushu City, and the positive rate of serum Echinococcus larvae antibodies in the population was 26.22% (134/511), and the detection rate of echinococcosis B-mode ultrasound was 1.37% (7/511). Among them, the detection rates of B-mode ultrasound for cystic echinococcosis (CE) and alveolar echinococcosis (AE) were 1.17% (6/511) and 0.20% (1/511), respectively. The positive rate of Echinococcus antigen in 543 canine feces detected by ELISA was 12.89% (70/543). PCR was used to test 497 canine feces, and the detection rate of Echinococcus was 3.02% (15/497). Among them, the detection rate of Echinococcus multilocularis was higher than that of Echinococcus granulosus [2.82% (14/497) vs 0.20% (1/497)], and the difference was statistically significant (χ 2 = 11.44, P < 0.001). No Echinococcus shiquicus was detected. Conclusions:The positive rates of Echinococcus larvae antibodies in the population and canine Echinococcus antigen in Yushu City, Qinghai Province are both relatively high. There is a mixed epidemic of CE and AE, with Echinococcus multilocularis being the main species.

Objective:Comparative analysis of the mNUTRIC and NRS-2002 scores for evaluating nutritional risk and predicting clinical outcomes in end stage liver disease patients.Method:A retrospective cohort study method was used to screen 114 cases with end-stage liver disease admitted to the intensive care unit (ICU) of the First Hospital of Lanzhou University from December 1, 2016 to March 31, 2021 according to the inclusion and exclusion criteria. The patient's demographic data, blood routine, blood biochemical indexes, coagulation function indexes, arterial blood gas analysis and imaging examination data were collected. The mNUTRIC score, NRS-2002 score, sequential organ failure (SOFA) score, model for end-stage liver disease (MELD) score, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, Child-Pugh grade, and clinical outcomes at 28 and 90 days at 24 h post-ICU admission were collected. The differences in clinical indicators between the mNUTRIC high group (≥5 points) and the low group, and the NRS-2002 high group (≥3 points) and the low group were compared. Spearman correlation analysis was used to explore the correlation between the mNUTRIC score and NRS-2002 score, clinical indicators, and 28 and 90-day mortality rates. Multivariate logistic regression analysis was used to determine the risk factors associated with 28-day and 90-day mortality in patients. The value of mNUTRIC score and NRS-2002 score in assessing the clinical outcomes of patients with end-stage liver disease was explored by receiver operating characteristic (ROC) curve.Results:The clinical indicators related to nutritional status of patients were worse in the high-mNUTRIC group than those in the low-mNUTRIC group, and the 28-day and 90-day mortality rates were significantly higher than those in the low-mNUTRIC group [89.0%(65/73) vs. 29.2%(12/41), 97.2%(71/73) vs. 39.0%(16/41), P<0.001]. There was no statistically significant difference in the incidence rate of hepatic encephalopathy, esophageal variceal bleeding, and ascites between the high and low mNUTRIC group. The clinical indicators related to nutritional status were worse in the high-NRS-2002 group than those in the low-NRS-2002 group of patients, and the 28-day and 90-day mortality rates were significantly higher than those in the low-group [73.0%(73/100) vs. 4/14, 81.0%(81/100) vs. 6/14, P=0.008, 0.004]. The NRS-2002 high-score group did not differ significantly from the low-score group in terms of hepatic encephalopathy, esophagogastric variceal bleeding, or ascites prevalence. Patient's age, white blood cell count (WBC), urea nitrogen (BUN), creatinine (UREA), uric acid (UA), total cholesterol (TG), Child-Pugh, MELD, SOFA, APACHE Ⅱscores were significantly positively correlated with the mNUTRIC score. Conversely, albumin (Alb) and Glasgow Coma Scale (GCS) were significantly negatively correlated. Patient's age, WBC, CREA, BUN, UREA, UA, Child-Pugh, MELD, SOFA, APACHE Ⅱwere significantly positively correlated with the NRS-2002 score.Conversely, albumin (Alb) and Glasgow Coma Scale (GCS) were significantly negatively correlated ( P<0.05). The 28-day and 90-day mortality rates of patients increased with the increase in the mNUTRIC scores. The mNUTRIC score was an independent predictor of death within 28 and 90 days in patients with end-stage liver disease. The area under the curve (AUC) of mNUTRIC for predicting patient death at 28 days was 0.864 (95% CI: 0.794-0.934). The AUC of NRS-2002 for predicting patient death at 28 days was 0.683 (95% CI: 0.573-0.792). The AUC of the two indicators combined for predicting patient death at 28 days was 0.868 (95% CI: 0.799-0.936). The AUC of mNUTRIC for predicting patient death at 90 days was 0.915 (95% CI: 0.861-0.969). The AUC of NRS-2002 for predicting patient death at 90 days was 0.715 (95% CI: 0.599-0.832). The AUC of the two indicators combined for predicting patient death at 90 days was 0.922 (95% CI: 0.871-0.972). Conclusion:mNUTRIC score and NRS-2002 score can better evaluate the nutritional status in patients with end-stage liver disease. The mNUTRIC score is a good predictor of 28-day and 90-day mortality in patients with end-stage liver disease, and its application value efficacy is enhanced when combined with NRS-2002.

Objective:To explore the safety and efficacy of TBF conditioning regimen of thiotepa, fludarabine and busulfan in haploidentical hematopoietic stem cell transplantation (haplo-HSCT) for recipients with hematological malignancies unsuitable for BUCY conditioning regimen (busulfan, cyclophosphamide & mBUCY).Method:A retrospective analysis was conducted for 20 recipients with malignant hematologic diseases receiving TBF conditioning regimen before haplo-HSCT at Soochow Hopes Hematologic Hospital from January 2020 to December 2023. The regimen-related toxicity of TBF was assessed by the Bearman scoring criteria. For comparing the safety and efficacy of TBF conditioning regimen with mBUCY regimen, propensity score matching was performed in a ratio of 1: 2 with disease type, patient age and gender as matching factors.Result:Mild oral mucositis and gastrointestinal reaction were major side-effects without severe cardiac events. Median time to neutrophil and platelet engraftments in TBF group was 11 and 18 days with comparable engraftment in mBUCY group. TBF regimen had a significantly lower incidence of grade Ⅱ-Ⅳ acute graft-versus-host disease (aGVHD) than mBUCY at Day 100 (5% vs 35%, P=0.01). No significant inter-group difference existed in overall survival (68% vs 62%, P=0.98) while 1-year incidence of graft-versus-host disease-free, relapse-free survival (GRFS) improved (63% vs 37%, P=0.06) in TBF group. Conclusion:TBF is a promising conditioning regimen with low toxicity and decent safety for haplo-HSCT. TBF patients tend to have a lower incidence of grade Ⅱ-ⅣaGVHD and better GRFS than mBUCY.

To explore the efficacy and safety of immunoadsorption (IA) in removing de novo donor specific antibody (DSA) after allogeneic hematopoietic stem cell transplantation (HSCT), the relevant clinical data were retrospectively reviewed for one female patient of severe aplastic anemia (SAA). Desensitization treatment with IA after HSCT was offered for removing de novo DSA and ultimately promoting platelet engraftment at First Affiliated Hospital of Soochow University in March 2021.

Objective To investigate the capillarization of liver sinusoidal endothelial cells (LSECs) and its association with hepatic fibrosis during the development of alveolar echinococcosis, so as to provide the basis for unraveling the mechanisms underlying the role of LSEC in the development and prognosis of hepatic injuries and hepatic fibrosis caused by alveolar echinococcosis. Methods Forty C57BL/6 mice at ages of 6 to 8 weeks were randomly divided into a control group and 1-, 2- and 4-week infection groups, of 10 mice in each group. Each mouse in the infection groups was intraperitoneally injected with 2 000 Echinococcus multilocularis protoscoleces, while each mouse in the control group was given an equal volume of phosphate-buffered saline using the same method. All mice were sacrificed 1, 2 and 4 weeks post-infection and mouse livers were collected. The pathological changes of livers were observed using hematoxylin-eosin (HE) staining, and hepatic fibrosis was evaluated through semi-quantitative analysis of Masson’s trichrome staining-positive areas. The activation of hepatic stellate cells (HSCs) and extracellular matrix (ECM) deposition were examined using immunohistochemical staining of α-smooth muscle actin (α-SMA) and collagen type I alpha 1 (COL1A1), and the fenestrations on the surface of LSECs were observed using scanning electron microscopy. Primary LSECs were isolated from mouse livers, and the mRNA expression of LSEC marker genes Stabilin-1, Stabilin-2, Ehd3, CD209b, GATA4 and Maf was quantified using real-time fluorescence quantitative PCR (qPCR) assay. Results Destruction of local liver lobular structure was observed in mice 2 weeks post-infection with E. multilocularis protoscoleces, and hydatid cysts, which were surrounded by granulomatous tissues, were found in mouse livers 4 weeks post-infection. Semi-quantitative analysis of Masson’s trichrome staining showed a significant difference in the proportion of collagen fiber contents in mouse livers among the four groups (F = 26.060, P < 0.001), and a higher proportion of collagen fiber contents was detected in mouse livers in the 4-week infection group [(11.29 ± 2.58)%] than in the control group (P < 0.001). Immunohistochemical staining revealed activation of a few HSCs and ECM deposition in mouse livers 1 and 2 weeks post-infection, and abundant brown-yellow stained α-SMA and COL1A1 were deposited in the lesion areas in mouse livers 4 weeks post-infection, which spread to surrounding tissues. Semi-quantitative analysis revealed significant differences in α-SMA (F = 7.667, P < 0.05) and COL1A1 expression (F = 6.530, P < 0.05) in mouse levers among the four groups, with higher α-SMA [(7.13 ± 3.68)%] and COL1A1 expression [(13.18 ± 7.20)%] quantified in mouse livers in the 4-week infection group than in the control group (both P values < 0.05). Scanning electron microscopy revealed significant differences in the fenestration frequency (F = 37.730, P < 0.001) and porosity (F = 16.010, P < 0.001) on the surface of mouse LSECs among the four groups, and reduced fenestration frequency and porosity were observed in the 1-[(1.22 ± 0.48)/μm² and [(3.05 ± 0.91)%] and 2-week infection groups [(3.47 ± 0.10)/μm² and (7.57 ± 0.23)%] groups than in the control group (all P values < 0.001). There was a significant difference in the average fenestration diameter on the surface of mouse LSECs among the four groups (F = 15.330, P < 0.001), and larger average fenestration diameters were measured in the 1-[(180.80 ± 16.42) nm] and 2-week infection groups [(161.70 ± 3.85) nm] than in the control group (both P values < 0.05). In addition, there were significant differences among the four groups in terms of Stabilin-1 (F = 153.100, P < 0.001), Stabilin-2 (F = 57.010, P < 0.001), Ehd3 (F = 31.700, P < 0.001), CD209b (F = 177.400, P < 0.001), GATA4 (F = 17.740, P < 0.001), and Maf mRNA expression (F = 72.710, P < 0.001), and reduced mRNA expression of Stabilin-1, Stabilin-2, Ehd3, CD209b, GATA4 and Maf genes was quantified in three infection groups than in the control group (all P values < 0.001). Conclusions E. multilocularis infections may induce capillarization of LSECs in mice, and result in a reduction in the expression of functional and phenotypic marker genes of LSECs, and capillarization of LSECs occurs earlier than activation of HSC and development of hepatic fibrosis.

Objective:To investigate the efficacy and safety of protein A immunoadsorption (PAIA) combined with rituximab (RTX) in highly sensitized patients who underwent haplo-hematopoietic stem cell transplantation (haplo-HSCT) .Methods:The clinical data of 56 highly sensitized patients treated with PAIA and RTX before haplo-HSCT at the First Affiliated Hospital of Soochow University and Soochow Hopes Hematonosis Hospital between March 2021 and June 2023 were retrospectively analyzed. The number of human leukocyte antigen (HLA) antibody types and the mean fluorescence intensity (MFI), humoral immunity, adverse reactions during adsorption, and survival within 100 days before and after adsorption were measured.Results:After receiving the PAIA treatment, the median MFI of patients containing only HLA Ⅰ antibodies decreased from 7 859 (3 209-12 444) to 3 719 (0-8 275) ( P<0.001), and the median MFI of HLA Ⅰ+Ⅱ antibodies decreased from 5 476 (1 977-12 382) to 3 714 (0-11 074) ( P=0.035). The median MFI of patients with positive anti-donor-specific antibodies decreased from 8 779 (2 697-18 659) to 4 524 (0–15 989) ( P<0.001). The number of HLA-A, B, C, DR, and DQ antibodies in all patients decreased after the PAIA treatment, and the differences were statistically significant (A, B, C, DR: P<0.001, DQ: P<0.01). The humoral immune monitoring before and after the PAIA treatment showed a significant decrease in the number of IgG and complement C3 ( P<0.001 and P=0.002, respectively). Forty-four patients underwent HLA antibody monitoring after transplantation, and the overall MFI and number of antibody types decreased. However, five patients developed new antibodies with low MFI, and nine patients continued to have high MFI. The overall survival, disease-free survival, non-recurrent mortality, and cumulative recurrence rates at 100 days post-transplantation were 83.8%, 80.2%, 16.1%, and 4.5%, respectively. Conclusions:The combination of PAIA and RTX has a certain therapeutic effect and good safety in the desensitization treatment of highly sensitive patients before haplo-HSCT.