Objective To explore the effects of S100 calcium-binding protein A9 (S100A9) gene knockout on the phenotype of systemic lupus erythematosus (SLE) in mice and to clarify the role of S100A9 in the pathogenesis of SLE. Methods Ten female C57BL/6 wild-type and S100A9 knockout (S100A9-KO ) mice were selected, with five wild-type and five S100A9-KO B6 mice receiving imiquimod (IMQ) cream to establish SLE mouse model. The other five wild-type and five S100A9-KO B6 mice were treated as control groups by wiping the skin of the right ear with a cotton swab. After 8 weeks, the mice were sacrificed. The serum was collected from each mouse to detect the levels of anti-double-stranded DNA (dsDNA) antibodies, immunoglobulin G (IgG), B cell activating factor (BAFF), and interleukin 6 (IL-6) using ELISA. The levels of serum creatinine were determined using a sarcosine oxidase method. Urine was collected to measure urinary protein concentration. Kidneys were collected and stained with hematoxylin and eosin (H&E) for evaluating histological changes. Results After IMQ treatment, the length and weight of spleen, levels of serum creatinine, anti-dsDNA antibodies, IgG, BAFF, IL-6, and urinary protein in the IMQ B6 group and IMQ S100A9-KO B6 group were significantly higher than those of the control groups. Lupus-like changes including increased glomerular volume and tubular epithelial swelling were observed in kidneys from the IMQ and IMQ S100A9-KO groups. However, compared with the IMQ B6 group, the IMQ S100A9-KO B6 group exhibited milder levels of serum and urine indicators as well as the lupus-like symptoms. Conclusion IMQ could induce lupus-like symptoms in both wild-type B6 mice and S100A9-KO B6 mice, but the lesions in S100A9 knockout mice are milder. Theses results suggested that S100A9 is involved in and promotes the pathogenesis of SLE.
Animals ; Lupus Erythematosus, Systemic/chemically induced* ; Female ; Calgranulin B/genetics* ; Mice, Knockout ; Mice, Inbred C57BL ; Phenotype ; Mice ; Interleukin-6/blood* ; Disease Models, Animal ; Antibodies, Antinuclear/blood* ; B-Cell Activating Factor/blood* ; Immunoglobulin G/blood* ; Kidney/pathology*

Objective To investigate the expression level and clinical application value of serum resistin in patients with systemic lu-pus erythematosus(SLE).Methods Forty-five SLE patients visited Nanjing Drum Tower Hospital,Clinical College of Nanjing Uni-versity of Chinese Medicine from January to August 2023 were enrolled in the study.The patients were scored and grouped according to the SLE disease activity index(SLEDAI),with SLEDAI＜9 score in the inactive group(n=32)and SLEDAI≥9 score in the active group(n=13).Thirty-four healthy individuals who underwent physical examination in our hospital were recruited as healthy controls.The clinical data and laboratory related indicators such as urine protein and serum complement C3 levels were collected from SLE pa-tients and healthy controls.Serum resistin levels were detected by enzyme-linked immunosorbent assay(ELISA).The clinical screening value of serum resistin for SLE was evaluated with the receiver operating characteristic(ROC)curve.The correlations of serum resistin levels with different laboratory indicators were determined by Pearson correlation analysis.Results The serum resistin levels in SLE patients([7.64±0.64]ng/mL)were significantly higher than that in healthy controls([2.56±0.43]ng/mL),and the difference was statistically significant(t=6.195,P＜0.01).The serum resistin levels in active SLE patients([10.10±1.45]ng/mL)were significant-ly higher than that in inactive SLE patients([6.64±0.60]ng/mL),and the difference was statistically significant(t=2.632,P＜0.05).The area under the ROC curve(AUCROC)of serum resistin for screening SLE was 0.897.When the cut-off value was 5.893 ng/mL,the sensitivity was 86.67%and the specificity was 82.35%.The serum resistin level in SLE patients was positively correlated with urine protein(r=0.692,P＜0.01),while negatively correlated with serum complement C3(r=-0.354,P＜0.05).Conclusion The expression levels of serum resistin in SLE patients are significantly increased and positively correlated with SLE disease activity and urine protein.Serum resistin may become a novel biomarker for the diagnosis and therapeutic effect assessment of SLE.

Objective:To investigate the effect of interleukin-12(IL-12)on liver lesions and oxidative stress pathway in Sjo-gren's syndrome mice,and to clarify the possible mechanism of liver lesions in Sjogren's syndrome.Methods:Saliva flow rate,liver function,liver pathology and IL-12 level were measured in NOD,IL-12 knockout(IL-12KO)NOD and C57BL/6(B6)mice.Different concentrations of IL-12 and JAK2,TYK2 inhibitors were applied to mouse hepatoma cells.The oxidative stress index of mouse serum and cell culture supernatants of each group were determined.Results:Compared with the other two groups of mice,the NOD mice had significantly higher GOT and GPT(P<0.05),and decreased serum GSH-PX,SOD and CAT(P<0.05).CAT,GSH,GSH-PX and SOD were decreased,while MDA was increased in mice treated with different concentrations of IL-12.JAK2/TYK2 inhibitors reversed regulatory effects of IL-12 on SOD,GSH and MDA in hepatoma cells(P<0.05).Conclusion:IL-12 may aggravated liver damage of Sj?gren's syndrome through promoting oxidative stress of hepatocytes.

SARS-CoV-2 is an emerging viral pathogen and a major global public health challenge since December of 2019, with limited effective treatments throughout the pandemic. As part of the innate immune response to viral infection, type I interferons (IFN-I) trigger a signaling cascade that culminates in the activation of hundreds of genes, known as interferon stimulated genes (ISGs), that collectively foster an antiviral state. We report here the identification of a group of type I interferon suppressed genes, including fatty acid synthase (FASN), which are involved in lipid metabolism. Overexpression of FASN or the addition of its downstream product, palmitate, increased viral infection while knockout or knockdown of FASN reduced infection. More importantly, pharmacological inhibitors of FASN effectively blocked infections with a broad range of viruses, including SARS-CoV-2 and its variants of concern. Thus, our studies not only suggest that downregulation of metabolic genes may present an antiviral strategy by type I interferon, but they also introduce the potential for FASN inhibitors to have a therapeutic application in combating emerging infectious diseases such as COVID-19.

Objective: To explore the plasma level change of soluble tumor necrosis factor related apoptosis inducing ligand (sTRAIL) in patients with systemic lupus erythematosus (SLE) and its clinical significance. Methods: Quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expressions of TRAIL and TRAIL receptors-1 (TRAIL-R1) and TRAIL-R2 in the peripheral blood mononuclear cells (PBMCs) derived from active SLE patients (n=26) and healthy controls. Enzyme linked immuno sorbent assay (ELISA) was used to detect the plasma level of sTRAIL in the active SLE patients (n=42), healthy controls (n=21). Pearson correlation analysis was used to analyze the correlation of sTRAIL with clinical and laboratory parameters. Results: The plasma levels of sTRAIL [(82±5) pg/ml] in SLE were significantly higher than that in healthy controls [(49±3) pg/ml], the difference was statistically significant (t=4.10, P<0.01). The plasma levels of sTRAIL in SLE with inactive disease [(92±14) pg/ml], mild active disease [(80±9) pg/ml], moderate active disease [(74±12) pg/ml] and severe active disease [(83±8) pg/ml] were higher than healthy controls, the difference was statistically significant (H=18.07, P<0.01). The mRNA levels of TRAIL and TRAIL-R2 in PBMCs derived from SLE patients were significantly higher than those in healthy controls [(1.04±0.08) vs (1.80±0.25), t=2.10, P<0.05 and (1.07±0.12) vs (2.08±0.21), t=3.27, P<0.01]. In SLE with moderate and severe active disease, plasma sTRAIL levels were associated with the 24 hours urine protein. Conclusion Plasma sTRAIL may be predictors of SLE disease activity and TRAIL pathway may be a new potential target of SLE treatment.

Objective To investigate the abnormality of intrahepatic biliary epithelial cells autophagy in primary biliary cirrhosis (PBC) mice,and explore the mechanism of UC-Mesenchymal stem cell (MSCs) in treating PBC.Methods After establishing the PBC model,we divided them into the PBC model group;the UC-MSCs treatment group and the Stattic group [Signal transducer and activator of transcription 3 (STAT3) inhibitor group].Six mice were used as the control group.Liver pathology and the serum pyruvate dehydrogenase complex E2 subunit (PDC-E2) antibody titers were detected.Autophagosome of the intrahepatic biliary epithelial cell was observed by electronic microscope.Protein levels of STAT3/pSTAT3,Beclin-1 were detected by western blot.We cultured human intrahepatic biliary epithelial cells in vitro,and down regulated STAT3.After stimulated by GCDC,we co-cultured them with UC-MSCs,and collected the cells in order to detect LC3 Ⅱ.The measurement data were compared with t test or single factor analysis of variance.Results Compared with the control group,periportal inflammatory cell infiltration and granuloma formation were observed in the PBC group.MSCs treatment decreased the infiltration of inflammatory cells.The level of antiPDC-E2 of the PBC group (107±18) ng/ml was higher than that of the control group (42±6) ng/ml (q=6.326,P＜0.01),MSCs treatment down regulated anti-PDC-E2 level (43±4) ng/ml (q=5.801,P＜0.01).More autophagosomes in the PBC group (5.00±1.29) than the control group (1.75±0.25) were observed (q=4.061,P＞0.05).Western blot showed that the level of Beclin-1 was higher in PBC group (1.80±0.36) than the control group (0.40±0.20) (q =6.757,P＜0.01),MSCs reduced the expression of Beclin-1 (0.86±0.06)(q=4.536,P＜0.05) as well as Stattic (0.72±0.03) (q=5.226,P＜0.05).PBC group had a higher expression level of STAT3 (1.80±0.42) (q=5.730,P＜0.05) and pSTAT3 (2.04±0.29)(q=6.492,P＜0.01) than the control group (0.50±0.05)(0.91±0.14).MSCs treatment decreased the expression of STAT3 (0.51±0.13)(q =5.703,P＜0.01) and pSTAT3 (0.76±0.07) (q =7.388,P＜0.01) in intrahepatic biliary epithelial cells.After down regulated STAT3 of HiBECs,MSCs reduced the expression of LC3 Ⅱ of HiBECs.Conclusion The intrahepatic biliary epithelial cells autophage of PBC mice is abnormal,MSCs can alleviate PBC by down regulating the autophage of intrahepatic biliary epithelial cells via STAT3.

Objective The purpose of this study is to observe the changes of immune cell subsets in lupus mice after umbilical cord mesenchymal stem cells (UC-MSCs) transplantation. Methods B6.MRL-Faslpr lupus mice were randomly divided into the following three groups: the UC-MSCs treated group, the fibroblast like synoviocytes (FLS) treated group and the untreated group. MSC (1×106) or FLS (1×106) were injected into the tail vein of lupus mice respectively. Four weeks after treatment, the spleen index was calculated. The pathological changes of kidney were assessed by HE staining. The frequencies of immune cell subsets in spleen and macrophage in kidney as well as abdominal cavity were analyzed by flow cytometry. Data were analyzed with t test. Results The spleen index of UC-MSCs treated lupus mice [(79 ±9) mg/10 g] and IgG level [(7.5±1.5) mg/ml] were significantly decreased when compared with FLS treated group [(147±23) mg/10 g, t=2.78, P<0.01] [(17.0 ±2.8) mg/ml, t=3.00, P<0.01] and the untreated group [(156 ±16) mg/10 g, t=4.29, P<0.01] [(16.7 ±1.6) mg/ml,t=4.01, P<0.01]. HE staining also showed that the pathological changes of kidney were alleviated after MSCs transplantation. In addition, the frequency of plasma cells in the untreated group [(2.61 2± 0.318)% vs (0.306±0.017)%, t=7.22, P<0.01] and the FLS treated group [(2.412±0.297)% vs (0.306±0.017)%, t=7.07, P<0.01] were markedly higher than MSCs treatment [(0.306 ±0.017)%]. Moreover, the frequency of CD25+Foxp3+/CD4+Treg in the MSCs treated group [(15.08±0.81)%] was significantly increased compared with the untreated group [(8.02 ±0.47)%, t=7.45, P<0.01] and FLS treated group [(8.80 ±0.23)%, t=7.39, P<0.01]. MSCs treatment resulted in a decrease in CXCR5+PD1+/CD4+Tfh and IFNγ+/CD4+Th1 subset, compared with the untreated group [(14.3±1.5)%vs (31.5±3.3)%, t=5.25, P<0.01] [(1.78±0.27)% vs (5.93±1.56)%, t=2.60, P<0.05] and the FLS treated group [(14.3±1.5)%vs (28.8±2.2)%, t=5.49, P<0.01] [(1.78±0.27)%vs (4.88±0.81)%, t=3.61, P<0.01]. The frequency of macrophage in kidney of the MSCs treated group [(3.52 ±0.37)%] was markedly increased compared with the untreated group[(1.58±0.29)%, t=3.25, P<0.01], while neither the IL4+/CD4+Th2 subset nor the IL17+/CD4+Th17subset and the frequency of macrophage in abdominal cavity showed significant changes in the three groups. Conclusion These findings suggest that the therapeutic effects of MSCs on lupus mice may mediate through increasing the frequency of spleen Treg and renal macrophage and decreasing the frequency of Tfh, Th1 and plasma cells.

OBJECTIVETo explore the molecular mechanism underlying the DEL phenotype among RhD negative ethnic Han individuals from Jiangsu, China.

METHODSThe DEL phenotype was determined by an adsorption elution test among 57 RhD negative blood donors. The Rh C, c, E, and e phenotypes were detected by a tube method. PCR with sequence-specific primering (PCR-SSP) assay was used to determine the RHCE genotypes. The RHD gene of the DEL individuals were amplified with polymerase chain reaction and subjected to Sanger sequencing analysis.

RESULTSAmong the 57 RhD negative donors, 10 (17.54%) were determined as having the DEL phenotype. The major RhCE phenotypes for DEL and RhD negative cases were RhCcee (80.0%) and Rhccee (61.7%), respectively. All RHD gene sequences of the 10 individuals have harbored a G>A mutation at position 1227 of exon 9.

CONCLUSIONA proportion of RhD negative individuals determined by routine serological method are actually DEL with RHD gene mutations. RHD *1227A is the most prevalent DEL genotype among ethnic Han Chinese from Jiangsu. Further research on the phenotype and underlying molecular mechanism of DEL is important for blood transfusion.

Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; Base Sequence ; Blood Donors ; China ; ethnology ; Exons ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Phenotype ; Polymorphism, Genetic ; Rh-Hr Blood-Group System ; genetics

Objective To explore the preventive effect of early umbilical cord mesenchymal stem cells (UC-MSCs) transplantation on MRL/lpr mice and the underly mechanisms.Methods Fourteen 10-week-old MRL/lpr mice were labeled and numbered.They were randomly divided into 2 groups by using random number table and injected with 1 ×106 UC-MSCs or PBS via tail vein respectively.Proteinuria was measured with Bradford method every 4 weeks.All mice were sacrificed at the age of 28 weeks, with the level of serum antidsDNA antibody and IL-17 detected by enzyme linked immunosorbent assay (ELISA).Splenic Th17 cells, as well as regulatory T cells (Treg) were examined by flow cytometry.Data were analyzed with t test and Pearson's correlation test.Results The onset of proteinuria was delayed for 4 weeks in UC-MSC-treated group compared with that in the control group.At the age of 28 weeks, the 24 hour proteinuria [(1.78±0.17) mg vs (4.77±0.98)mg, t=2.99, P＜0.05] and the spleen weight [(0.149±0.009) g vs (0.273±0.052) g, t=2.33, P＜0.05] in UC-MSCtreated group were significantly lower than those in the control group.There was also a trend of the decline of serum anti-dsDNA antibody and IL-17 level after UC-MSCs transplantation.Compared with those in the control group, both the percentage and the absolute number of Th17 cells were significantly decreased in UC-MSC-treated group [(0.90±0.19)％ vs (2.81±0.50)％, t=3.54, P＜0.01 and (3.7±0.8)×105 vs (19.3±3.7)×105, t=4.12,P＜0.01].Meanwhile, the percentage of Treg elevated after UC-MSCs treatment.The ratio of Th17/Treg was significantly lower in UC-MSC-treated group than that in the control group (0.11±0.03 vs 0.50±0.09, t=4.23,P＜0.01).Both the ratio of Th17/Treg (r=0.73, P＜0.01;r=0.59, P＜0.05) and serum IL-17 level (r=0.78, P＜0.01;r=0.56, P＜0.05) was positively correlated with the level of 24 hour proteinuria and anti-dsDNA antibody respectively in MRL/lpr mice.Conclusion Early UC-MSCs transplantation helps to delay disease onset and ameliorate disease progression in MRL/lpr mice, which may act through the modulation of Th17/Treg balance.

Objective Whether the bone marrow cells (BMC) derived from systemic lupus erythematosus (SLE) could transmit autoimmune disease was studied for the purpose of clarifying the role of BMC in SLE pathogenesis.The effects of bone marrow mesenchymal stem cells (MSC) from SLE and control mice on the SLE BMC-induced symptoms were compared to elucidate the role of MSC in SLE.Methods Six-week-old B6.MRL-Fas mice were randomly divided into 3 groups.One group was transplanted with BMC from the 30-week-old B6.MRL-Faslg mice.One group was co-transplanted with BMC from the 30-week-old B6.MRL-Fasr mice and bone marrow MSC from the age-matched B6.MRL-Faslpr mice.One group was co-transplanted with BMC from the 30-week-old B6.MRL-Faslg mice and bone marrow MSC from the age-matched C57BL/6 mice.Before transplantation,the recipient mice received irradiation by an X-ray source.The levels of serum antinuclear antibody (ANA) and proteinuria were measured with enzyme linked immunosorbent assay (ELISA) and Bradford method every 4 weeks,respectively.The survival rate was recorded.All mice were sacrificed 18 weeks later.Splenic plasma cells,Th1,Th2 and Th17 cells were measured by flow cytometry.Statistical analyses were performed using the independent t test and ANOVA.Results Eight weeks after transplantation,ANA was positive in all the recipient mice.However,there was no significant difference between the three groups (P＞0.05).No proteinuria was observed in all the recipient mice.The mice received BMC from the 30-week-old B6.MRL-Fasr mice and bone marrow MSC from the age-matched B6.MRL-Fasr mice showed an elevated trend of the percentages of splenic plasma cells,Th1,Th2 and Th17 cells compared with the other two groups,plasma cells [(1.05±0.16)％,(0.58±0.11)％,t=2.53,P＞0.05;(1.05±0.16)％,(0.71±0.18)％,t=1.45,P＞0.05],Th1 cells [(6.6±2.2)％,(5.7±1.0)％,t=0.38,P＞0.05;(6.6±2.2)％,(4.0±1.7)％,t=0.96,P＞0.05],Th2 cells [(3.3±0.4)％,(2.1±0.6)％,t=1.76,P＞0.05;(3.3±0.4)％,(2.2±0.6)％,t=1.51,P＞0.05],Th17 cells [(2.30±0.71)％,(1.31±0.31)％,t=1.27,P＞0.05;(2.30±0.71)％,(1.12±0.27)％,t=1.67,P＞0.05].However,there was no significant difference between the groups.The survival rate of the three groups was 43％,43％ and 80％ respectively.And the survival rate of the mice received BMC from the 30-week-old B6.MRL-Fasr mice and bone marrow MSC from the age-matched C57BL/6 mice was significantly higher than those of the other groups.Conclusion Our results indicate that BMC from SLE can transmit autoimmune disease.The bone marrow MSC can not prevent lupus-like presentations induced by BMC from SLE.Transplantation of bone marrow MSC from C57BL/6 mice can significantly elevate the survival rate.