Male patients with primary Sjögren's syndrome have unique clinical manifestations and circulating lymphocyte profiles.

OBJECTIVES: We aimed to explore the relationship between clinical characteristics and circulating lymphocyte profiles in Chinese male patients with primary Sjögren's syndrome (pSS). METHOD: Data from 397 patients with pSS were analyzed retrospectively. 37 were male, which is a prevalence of 9.3%. The clinical, laboratory, and immunophenotypic profiles of peripheral blood lymphocyte subsets were compared between male and female pSS patients. RESULTS: Male patients with primary Sjögren's syndrome have unique clinical manifestations and circulating lymphocyte profiles. Male patients complained more about xerophthalmia and presented with more extra-glandular manifestations as compared with female patients. The CD4+/CD8+ ratio (P = 0.030), the prevalence of CD4-CD8- T cells in lymphocytes (P = 0.020), the absolute number of CD4-CD8- T cells (P = 0.035), the prevalence of CD4+ T cells in lymphocytes (P < 0.001), and the absolute number of CD4+ T cells (P = 0.023) were significantly lower in male patients compared to female patients. On the other hand, the prevalence of CD8+CD28+ T cells (P = 0.030) and CD4+CD25high T cells (P = 0.040) in lymphocytes was significantly higher in male patients than in female patients. Moreover, compared to females with pSS, an elevated serum IgG level, low C3 and C4 levels, anti-SSB positivity, and ANA titers of ≥ 1:160 positivity were more frequent in male with pSS. CONCLUSIONS: Male patients with pSS have distinctive peripheral blood lymphocyte subpopulations, present with more severe clinical symptoms and immunological features, and have an unfavorable prognosis. Key Points • Male patients with pSS have more severe clinical symptoms and specific characteristics of peripheral blood lymphocyte subsets. • Male pSS patients exhibit a higher intensity of the disease (as evaluated by ESSDAI). • Male patients with pSS require individualized treatment regimens and closer follow-up.

Phenotypic and Genetic Analysis of KPC-49, a KPC-2 Variant Conferring Resistance to Ceftazidime-Avibactam and Maintaining Resistance to Imipenem and Meropenem.

Purpose: Klebsiella pneumoniae, a gram-negative bacterium, poses a severe hazard to public health, with many bacterial hosts having developed resistance to most antibiotics in clinical use. The goal of this study was to look into the development of resistance to both ceftazidime-avibactam and carbapenems, including imipenem and meropenem, in a K. pneumonia strain expressing a novel K. pneumoniae carbapenemase-2 (KPC-2) variant, referred to as KPC-49. Methods: After 1 day of incubation of K1 on agar containing ceftazidime-avibactam (MIC = 16/4 mg/L), a second KPC-producing K. pneumoniae strain (K2) was recovered. Antimicrobial susceptibility assays, cloning assays, and whole genome sequencing were performed to analyse and evaluate antibiotic resistance phenotypes and genotypes. Results: K. pneumoniae strain (K1), that produced KPC-2, was susceptible to ceftazidime-avibactam but resistant to carbapenems. The K2 isolate harboured a novel bla KPC-49 variant, which differs from bla KPC-2 by a single nucleotide (C487A), and results in an arginine-serine substitution at amino acid position 163 (R163S). The mutant K2 strain was resistant to both ceftazidime-avibactam and carbapenems. We demonstrated the ability of KPC-49 to hydrolyse carbapenems, which may be attributed to high KPC-49 expression or presence of an efflux pump and/or absence of membrane pore proteins in K2. Furthermore, blaKPC-like was carried on an IncFII (pHN7A8)/IncR-type plasmid within a TnAs1-orf-orf-orf-orf-orf-orf-ISKpn6-bla KPC-ISKpn27 structure. The bla KPC-like gene was flanked by various insertion sequences and transposon elements, including the Tn3 family transposon, such as TnAs1, TnAs3, IS26, and IS481-ISKpn27. Conclusion: New KPC variants are emerging owing to sustained exposure to antimicrobials and modifications in their amino acid sequences. We demonstrated the drug resistance mechanisms of the new mutant strains through experimental whole genome sequencing combined with bioinformatics analysis. Enhanced understanding of laboratory and clinical features of infections due to K. pneumoniae of the new KPC subtype is key to early and accurate anti-infective therapy.

"Typhoidal Cells" Appear in a Woman with Hemophagocytic Syndrome Secondary To Brucellosis: A Case Report.

We report a case of hemophagocytic syndrome (HPS) secondary to brucellosis, in which typhoidal cells were found in bone marrow, suggesting typhoidal cells present not only in Salmonella typhi infections but also in other bacterial infections. Typhoidal cells in bone marrow can be used to quickly identify the presence of bacterial infection pending the results of bone marrow and/or blood cultures.

Anticancer activities of natural antimicrobial peptides from animals.

Cancer is the most common cause of human death worldwide, posing a serious threat to human health and having a negative impact on the economy. In the past few decades, significant progress has been made in anticancer therapies, but traditional anticancer therapies, including radiation therapy, surgery, chemotherapy, molecular targeted therapy, immunotherapy and antibody-drug conjugates (ADCs), have serious side effects, low specificity, and the emergence of drug resistance. Therefore, there is an urgent need to develop new treatment methods to improve efficacy and reduce side effects. Antimicrobial peptides (AMPs) exist in the innate immune system of various organisms. As the most promising alternatives to traditional drugs for treating cancers, some AMPs also have been proven to possess anticancer activities, which are defined as anticancer peptides (ACPs). These peptides have the advantages of being able to specifically target cancer cells and have less toxicity to normal tissues. More and more studies have found that marine and terrestrial animals contain a large amount of ACPs. In this article, we introduced the animal derived AMPs with anti-cancer activity, and summarized the types of tumor cells inhibited by ACPs, the mechanisms by which they exert anti-tumor effects and clinical applications of ACPs.

Identification of Isthmin1 in the small annual fish, Nothobranchius guentheri, as a novel biomarker of aging and its potential rejuvenation activity.

Isthmin 1 (Ism1) has been shown to play roles in multiple biological processes including morphogenesis, hematopoiesis, antiviral immune response and suppression of tumor growth. However, it remains unknown if it plays any role in aging process. Here we showed for the first time that Ism1 was a new age-related biomarker, which decreased with age in fish, mice and humans. Interestingly, Ism1 was also useful to measure the "rejuvenated" age of fish Nothobranchius guentheri reversed by salidroside treatment and temperature reduction, providing additional evidence that Ism1 was an aging biomarker. In addition, we clearly showed that dietary intake of recombinant Ism1 had little effects on the body length and weight of aging N. guentheri, but it retarded the onset of age-related biomarkers and prolonged both the maximum and median lifespan of the fish. We also showed that Ism1 exerted its rejuvenation activity via the enhancement of antioxidant system. Collectively, our results indicate that Ism1 is not only is a novel biomarker of aging but also a potential rejuvenation factor capable of reversing aging of N. guentheri.

Holliday junction­recognition protein modulates apoptosis, cell cycle arrest and reactive oxygen species stress in human renal cell carcinoma.

Holliday junction recognition protein (HJURP) is involved in the regulation of mortality in various cell types, including renal cell carcinoma (RCC) cells. The specific mechanisms by which HJURP regulates RCC cell apoptosis and the cell cycle have not been previously investigated, to the best of our knowledge. In the present study, the expression of HJURP in RCC tissues and adjacent paracancerous renal tissue, as well as in RCC cell lines, was analyzed using reverse transcription­quantitative PCR and western blot analysis. The A498 RCC cells were transfected with an HJURP overexpression vector, which resulted in reduced proliferation, as demonstrated using immunofluorescence staining, a Cell Counting Kit­8 assay and a colony formation assay. Flow cytometry and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labelling assays were used to determine the effect of HJURP on the cell cycle and apoptosis of RCC cells. Proteins associated with the reactive oxygen species (ROS) status were analyzed using western blot analysis. The expression of HJURP was lower in RCC tissues and cells compared with that in the adjacent paracancerous renal tissues and control cells. Furthermore, overexpression of HJURP resulted in a decrease in cell viability and proliferation in vitro. Overexpression of HJURP resulted in cell cycle arrest at the G0/G1 phase, cell apoptosis and an increase in ROS stress. In addition, the phosphorylated/total sirtuin 1 (SIRT1) protein ratio was decreased, whereas the expression of peroxisome proliferator­activated receptor (PPAR)γ was increased in the HJURP­overexpressing RCC cells. In clinical practice, decreased HJURP expression may be associated with poor prognosis in patients with RCC. These results suggest that HJURP may regulate cell apoptosis and proliferation in RCC cells and this may be mediated by PPARγ/SIRT1. Thus, HJURP may be used as a predictor of prognosis in patients with RCC.

Enhancement of adaptive immune responses of aged mice by dietary intake of ß-glucans, with special emphasis on anti-aging activity.

The naturally occurring polysaccharide, ß-1,3-glucans, a well-known immunostimulant, has been highly valued for many years for their health-promoting and anti-aging properties, but its mode of action is poorly understood. In this study, we first showed that oral administration of ß-1,3-glucans did not affect the general condition and physiology of male mice throughout the trial period. We then showed that dietary intake of ß-1,3-glucans induced a significant increase in T helper cells (CD4+) in young, middle-aged and aged male mice. We also showed that ß-1,3-glucans supplementation considerably increased the delayed-type hypersensitivity (DTH) response, a T cell-mediated immune response, in young and aged mice. In addition, we found that ß-1,3-glucans supplementation remarkably promoted the production of total anti-keyhole limpet hemocyanin (KLH) immunoglobulin G (IgG), anti-KLH IgG1, and anti-KLH IgG2a in young and aged mice without disturbing immune homeostasis. These data together indicate that oral administration of ß-1,3-glucans enhanced the adaptive immune responses of aged mice without disturbing their general condition and physiology, supporting the idea that ß-1,3-glucans are capable of counteracting the immunosenescence in mice. They also suggest that ß-1,3-glucans can be clinically useful to help the elderly generate an improved response to vaccine with stronger humoral and cell-mediated immune responses.

Dietary Intake of ß-Glucans Can Prolong Lifespan and Exert an Antioxidant Action on Aged Fish Nothobranchius guentheri.

One of the widely accepted conjectures regarding mechanisms of aging is probably the oxidative stress hypothesis. ß-1,3-Glucans, well-known immunostimulants, have been shown to increase nonspecific immunity and resistance against infections or pathogenic bacteria in several fish species, but its antiaging function remains poorly understood. By feeding of ß-1,3-glucans to the annual fish, Nothobranchius guentheri, we detected the survivorship of the fish and estimated the development of age-related biomarkers at different stages. We first showed that administration of ß-1,3-glucans was able to prolong the lifespan of the fish (p < 0.05). We then showed that ß-1,3-glucans clearly reduced the accumulation of lipofuscin in the gills and the senescence-associated ß-galactosidase in the caudal fins. Moreover, ß-1,3-glucans were able to lower the levels of protein oxidation, lipid peroxidation, and reactive oxygen species (ROS) in the muscles. Finally, ß-1,3-glucans could promote the activities of the antioxidant enzymes, including catalase, superoxide dismutase, and glutathione peroxidase in the fish, and slow down the increase of P66shc, a critical factor involved in the regulation of intracellular ROS contents. These data together suggest for the first time that ß-1,3-glucans can extend the lifespan, delay the onset of age-related biomarkers and exert an antioxidant action of the aged fish, N. guentheri. It also implies that ß-1,3-glucans may be potentially useful for health care in the elderly, including extension of the lifespan.

Dyslipidemia in patients with systemic lupus erythematosus: Association with disease activity and B-type natriuretic peptide levels.

The aim of the present study was to evaluate the association between the levels of lipids and B-type natriuretic peptide (BNP) in systemic lupus erythematosus (SLE) patients with heart failure (HF). A total of 46 patients with active SLE and 40 healthy, age-matched control subjects were studied. BNP was measured by an immunofluorescence assay in fresh plasma. Total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol, apolipoprotein (Apo) B, ApoA-I and lipoprotein(a) were assessed. Compared with the control subjects, HDL-C and ApoA-I levels were considerably decreased and TG level increased markedly from SLE patients. The average concentration of HDL-C and ApoA-I in the SLE group with HF was significantly reduced compared to those patients without HF. The results showed that the levels of HDL-C and ApoA-I in SLE patients were negatively correlated with BNP. Disease activity was associated with the TC and TG levels. The present data indicated the presence of a cardiovascular (CV) risk in active SLE with high disease activity, which was demonstrated by the high frequency of dyslipidemia and higher BNP concentrations. Therefore, dyslipoproteinemia may underlie some of the increased risk for CV disease and HF in patients with SLE.

Altered TNFAIP3 mRNA expression in peripheral blood mononuclear cells from patients with rheumatoid arthritis.

The tumor necrosis factor α-induced protein-3 (TNFAIP3) gene functions in negative immunoregulation and its single-nucleotide polymorphisms (SNPs) are associated with rheumatoid arthritis (RA) disease. However, its expression level in immune cells from RA patients remains unclear. The aim of the present study was to investigate whether the expression of TNFAIP3 is changed in patients with RA. Reverse transcription-quantitative polymerase chain reaction analysis was used to determine TNFAIP3 mRNA expression in peripheral blood mononuclear cells (PBMCs) from patients with RA and healthy controls. TNFAIP3 expression was decreased in RA patients compared with the healthy controls. The expression level of the TNFAIP3 gene negatively correlated with the RA score, anti-cyclic citrullinated peptide (CCP) antibody levels and C-reactive protein levels. Furthermore, RA patients with positive results of anti-CCP antibodies had a lower expression of TNFAIP3 than those without anti-CCP antibodies. In conclusion, the present results suggest that the insufficient expression of the TNFAIP3 gene in PBMCs may correlate with the diagnosis of RA.

Altered glutamate cysteine ligase expression and activity in renal cell carcinoma.

Oxidative stress has been linked to the progression of mutations and cancer. Increased glutathione (GSH) contents have been observed in a number of different human cancer tissues. GSH is synthesized de novo in a two-step process catalyzed by glutamate cysteine ligase (GCL). The present study aimed to investigate whether GCL was associated with renal cell carcinoma (RCC). The protein expression levels of the GCL subunits (catalytic subunit, GCLc; and modulatory subunit, GCLm) and GCL activity were examined in renal cancer tissue. A total of 46 patients fulfilling the RCC criteria of the World Health Organization, revised in 2004, were enrolled. The tumor and adjacent tissues were sampled from all the subjects by surgery. The study demonstrated that GCLc and GCLm protein expression and the GCL activity were significantly increased in the tumor tissue from RCC patients. These results indicate that increased expression and enzymatic activity of GCL is closely associated with RCC and thus, this suggests an important role for GSH in the pathogenesis of RCC.

Altered glutamate cysteine ligase activity in peripheral blood mononuclear cells from patients with systemic lupus erythematosus.

Reductions in glutathione (GSH) levels have been shown to be associated with aging and the pathogenesis of a variety of diseases, including systemic lupus erythematosus (SLE). Glutamate cysteine ligase (GCL) catalyzes the first and rate-limiting step of GSH synthesis. In order to appraise the correlation between oxidative stress and the severity and activity of SLE, GSH, oxidized GSH (GSSG) and thioredoxin (TRX) concentrations and the enzymatic activity levels of GCL in peripheral blood mononuclear cells (PBMCs) from patients with SLE and healthy controls were studied. In patients with SLE, the levels of GCL activity and GSH decreased, while TRX and GSSG levels increased when compared with those in the healthy controls. GSH concentrations and GCL activity levels negatively correlated with the SLE disease activity index and erythrocyte sedimentation rate. Furthermore, patients with SLE and nephritis had lower levels of GSH and GCL activity and higher levels of TRX and GSSG compared with those in SLE patients without nephritis. Therefore, the results of the present study indicate that insufficient levels of GSH and GCL activity in PBMCs may contribute to the pathogenesis of SLE.

Resveratrol affects the expression of glutamate cysteine ligase in the kidneys of aged rats.

The aim of the present study was to evaluate the effect that a dietary intake of resveratrol (RSV) had on the expression of glutamate cysteine ligase (GCL) in the kidneys of aged rats. Young, middle-aged and aged rats were each randomly divided into two groups. The control groups were fed a controlled diet and the experimental groups received a controlled diet supplemented with RSV. GCL activity levels in the kidneys were determined. Protein content and relative gene expression levels of the two subunits of GCL were evaluated by western blot analysis and quantitative polymerase chain reaction, respectively. GCL activity levels significantly increased in the kidneys of aged rats fed the RSV-supplemented diet. In addition, RSV markedly increased the protein content and relative mRNA expression levels of the GCL subunits in the kidneys of aged rats. These observations have important implications for the development of therapeutic agents for the kidneys that may enable the elderly population to combat oxidative stress.

Rejuvenating activity of salidroside (SDS): dietary intake of SDS enhances the immune response of aged rats.

It is well known that immune response decreases with aging. Salidroside (SDS), an antioxidant component isolated from the traditional Chinese medicine roseroot Rhodiola rosea, has been demonstrated to possess potent anti-aging and health-promoting activities. However, the mechanism underlying these activities is poorly understood. In this study, we clearly demonstrated that (1) dietary intake of SDS induced a considerable increase in total T cells (CD3(+)) and T helper cells (CD4(+)) in aged (21 months old) Wistar male rats; (2) SDS supplementation significantly increased the DTH response, a T cell-mediated immune response, in aged rats; and (3) SDS supplementation remarkably promoted the production of total anti-KLH IgG, anti-KLH IgG1, and anti-KLH IgG2α in aged rats without disturbing immune homeostasis. These indicate that SDS is able to counteract immunosenescence, thereby resulting in rejuvenation. Practically, SDS may be used to help the elderly to generate an improved response to vaccine with stronger humoral and cell-mediated immune responses.

Dietary intake of resveratrol enhances the adaptive immunity of aged rats.

It is well known that immune response declines with aging. Resveratrol, a polyphenol that occurs naturally in several plant species including grapevines and berries, has been shown to have potent antiaging and health-promoting activities. However, the mechanism underlying these activities remains largely unknown. Here we clearly demonstrate that: (1) Dietary intake of resveratrol induced a significant increase in T helper cells (CD4(+)) in middle-aged (12 months old) and aged (21 months old) Wistar male rats; (2) resveratrol supplementation considerably increased the delayed-type hypersensitivity response, a T cell-mediated immune response, in aged rats; and (3) reveratrol supplementation remarkably promoted the production of total anti-keyhole limpet hemocyanin (KLH) immunoglobulin G (IgG), anti-KLH IgG(1), and anti-KLH IgG(2α) in aged rats without disturbing immune homeostasis. These data together indicate that resveratrol is capable of counteracting immunosenescence, thereby leading to rejuvenation. In practice, resveratrol can be useful to help the elderly generate an improved response to vaccine with stronger humoral and cell-mediated immune responses.

[Relationship between polymorphism sites of IRF5, TLR-9 and SLE patients in Shandong Han population].

OBJECTIVE: To investigate two single nucleotide polymorphism sites of IRF5 and TLR-9 and to detect their relationship with SLE (systemic lupus erythematosus) in a Han population from Shandong province. METHODS: The polymorphisms of rs2004640 G/T, rs10954213 G/A in IRF-5 and rs187084C/T, rs352139A/G in TLR-9 were detected with PCR-RFLP in 92 cases of SLE and 88 healthy controls. The genotype and allele frequencies were calculated and analyzed. RESULTS: (1) The genotype frequencies of GG, GT and TT in rs2004640 site in SLE were 0.198, 0.521 and 0.281 respectively. The difference was significant between SLE and controls (chi(2) = 8.73, P < 0.05); the genotype frequencies of GG, GA and AA at rs109542130 site in SLE were 0.318, 0.409 and 0.273 respectively. The difference was significant between SLE and controls (chi(2) = 6.36, P < 0.05). (2) The genotype frequencies of CC, CT and TT at rs187084 site in SLE were 0.185, 0.413 and 0.402 respectively. There was no difference between SLE and controls (chi(2) = 2.99, P > 0.05); the genotype frequencies of AA, AG and GG at rs352139 site in SLE were 0.228, 0.533 and 0.239 respectively. There was no difference between SLE and controls (chi(2) = 4.54, P > 0.05). CONCLUSION: The polymorphism of rs2004640 and rs10954213 in IRF5 may be associated with SLE in the population of Han nationality from Shandong province. However, the polymorphism of rs187084 and rs352139 in TLR-9 is not associated with SLE.

[Establishment of the transfected cell line expressing the mouse OX40 gene and its potency to induce differentiation of B cells].

AIM: To establish HUVECs line expressing mouse OX40(CD134) and to study its promotive effect on proliferation of B cells. METHODS: The cDNA fragment encoding mouse OX40 was obtained from the total RNA of ConA-activated lymphocytes of thymus by using RT-PCR and cloned into pUCm-T vector. The cDNA was then inserted into the eukaryotic expression vector pIRES2-EGFP. The recombinant vector was transfected into HUVECs with lipofectin reagent, and the positive cellular clones were selected by G418. Expression of mouse OX40 in the transfected cells was analyzed by FCM. The differentiation of B cells in vitro induced by OX40 signal was studied by means of (3)H-TdR method. RESULTS: The cDNA fragment in the recombinant plasmid was consistent with the reported mouse OX40 cDNA in GenBank, which was confirmed by DNA sequencing, PCR and enzyme digestion. The HUVECs stably expressing the mouse OX40 were successfully cloned. The transfected cells promoted the differentiation of B cells in vitro and there existed a synergic effect between OX40/OX40L and CD40/CD40L signals. CONCLUSION: Transfected cell line expressing the mouse OX40 gene is established successfully. OX40 enhances the proliferation of B cells.