Prospects for the Application of Transplantation With Human Amniotic Membrane Epithelial Stem Cells in Systemic Lupus Erythematosus.

Systemic lupus erythematosus (SLE) is a multi-organ and systemic autoimmune disease characterized by an imbalance of humoral and cellular immunity. The efficacy and side effects of traditional glucocorticoid and immunosuppressant therapy remain controversial. Recent studies have revealed abnormalities in mesenchymal stem cells (MSCs) in SLE, leading to the application of bone marrow-derived MSCs (BM-MSCs) transplantation technique for SLE treatment. However, autologous transplantation using BM-MSCs from SLE patients has shown suboptimal efficacy due to their dysfunction, while allogeneic mesenchymal stem cell transplantation (MSCT) still faces challenges, such as donor degeneration, genetic instability, and immune rejection. Therefore, exploring new sources of stem cells is crucial for overcoming these limitations in clinical applications. Human amniotic epithelial stem cells (hAESCs), derived from the eighth-day blastocyst, possess strong characteristics including good differentiation potential, immune tolerance with low antigen-presenting ability, and unique immune properties. Hence, hAESCs hold great promise for the treatment of not only SLE but also other autoimmune diseases.

Reporting guidelines for traditional Chinese medicine could be improved: a cross-sectional study.

OBJECTIVES: The aim of this study is to identify available reporting guidelines for traditional Chinese medicine (TCM), delineate their fundamental characteristics, assess the scientific rigor of their development process, and evaluate their dissemination. STUDY DESIGN AND SETTING: A search was conducted in Medline (via PubMed), China National Knowledge Infrastructure (CNKI), SinoMed, WANFANG DATA, and the EQUATOR Network to identify TCM reporting guidelines. A preprepared Excel database was used to extract information on the basic characteristics, development process, and dissemination information. The development process quality of TCM reporting guidelines was assessed by evaluating their compliance with the Guidance for Developers of Health Research Reporting Guidelines (GDHRRG). The extent of dissemination of these guidelines was analyzed by examining the number of citations received. RESULTS: A total of 26 reporting guidelines for TCM were obtained from 20 academic journals, with 61.5% of them published in English journals. Among the guidelines, 14 (53.8%) were registered in the EQUATOR Network. On average, the compliance rate of GDHRRG guidelines was reported to be 63.3% ranging from 22.2% to 94.4%. Three steps showed poor compliance, namely guideline endorsement (23.1%), translated guidelines (19.2%), and developing a publication strategy (19.2%). Furthermore, the compliance rate of GDHRRG guidelines published in English journals was higher than that in Chinese journals. In terms of the dissemination, 15.4% of the guidelines had been cited over 100 times, while 73.1% had been cited less than 50 times. CONCLUSION: The development of TCM reporting guidelines still has limitations in terms of regarding scientific rigor and follow-up dissemination. Therefore, it is important to ensure adherence to the scientific process in the development of TCM reporting guidelines and to strengthen their promotion, dissemination, and implementation.

Toll-Like Receptor 4 Deficiency Ameliorates Propofol-Induced Impairments of Cognitive Function and Synaptic Plasticity in Young Mice.

Propofol is one of the most used intravenous anesthetic agents, which is widely used in clinical anesthesia induction and maintenance of pediatric patients. Exposure of the developing brain to propofol has been reported to lead to adverse brain changes, which in turn can induce persistent behavioral abnormalities in adulthood. However, the mechanisms by which propofol exposure in the developing brain induces cognitive impairment remain unclear. Here we report that repeated propofol exposure during the second postnatal week impairs spatial learning and memory in young mice. The reduced excitatory synaptic function and synaptogenesis in hippocampal CA1 neurons underlie this cognitive impairment. Propofol exposure specifically activates Toll-like receptor 4 (TLR4)-myeloid differentiation primary response protein 88 (MyD88)-NF-κB signaling pathway. TLR4 deficiency recues propofol exposure-induced synaptic function and cognitive deficits in young mice. Thus, we provide evidence that the activation of the TLR4-mediated pathway by propofol exposure may serve as a crucial trigger for the cognitive impairment in young adulthood caused by repeated exposure to propofol in the developing brain.

Variations in Macrophage Activation Syndrome-associated Cardiac Diseases: A Report on Two Cases.

Macrophage activation syndrome (MAS), a secondary hemophagocytic lymphohistiocytosis characterized by an excessive systemic inflammatory response, is a life-threatening and rare disease. Cardiovascular damage is a common and severe complication of the disease, however, it is easily ignored and not well studied. Herein, we report two cases of patients with MAS-associated heart damage and review the clinical characteristics, mechanism, and treatment. Case 1 along with systemic lupus erythematosus and Kikuchi necrotizing lymphadenitis occurred in fatal acute heart failure, and case 2 complicated adult-onset Still's Disease began with atrial fibrillation and had some improvement with the treatment of high dose corticosteroids. MAS-associated heart damage is a critical issue in clinical settings, and the etiology and mechanisms of MAS-associated cardiovascular diseases are likely multifactorial. The manifestations were various and high levels of the cytokines and cardiac damage may contribute to poor prognosis. Therefore, early intensive immunosuppressive therapy probably improves the treatment outcome.

Paeoniflorin Inhibits LPS-Induced Activation of Splenic CD4+ T Lymphocytes and Relieves Pathological Symptoms in MRL/lpr Mice by Suppressing IRAK1 Signaling.

Interleukin-1receptor-associated kinase 1 (IRAK1) plays a critical role in systemic lupus erythematosus (SLE). It was reported that SLE was associated with an inflammatory response mediated by defective immune tolerance, including overproduction of autoantibodies, chronic inflammation, and organ damage. Previous reports stated paeoniflorin (PF) had an immunosuppressive effect. The purpose of this study was to determine the anti-inflammatory effect of PF in SLE and its underlying mechanisms. Followed by induced with lipopolysaccharide (LPS), the splenocytes and the isolated CD4+ T lymphocytes of MRL/lpr mice were divided into three groups: control group, LPS group, and LPS + PF group, respectively. MRL/MP mice were used as the control group (treated with distilled water). The MRL/lpr mice were randomly divided into three groups: the model group (treated with distilled water), the prednisone group, and the PF group. The MRL/lpr mice were treated with prednisone acetate (5 mg/kg) and PF (25, 50, and 75 mg/kg) for eight weeks. Subsequently, ELISA, qRT-PCR, western blotting, HE, and Masson staining were performed to detect various indicators. The results of Cell Counting Kit-8 (CCK-8) showed that 10 µg/mL of LPS had the optimum effect on cell viability, and 50 µmol/L of PF had no obvious cytotoxicity to LPS-treated cells. PF reduced the expression level of IRAK1-nuclearfactor-κB (NF-κB) and its downstream inflammatory cytokines in the splenocytes and CD4+ T lymphocytes of MRL/lpr mice stimulated by LPS, especially in the latter. The serum antibody contents in the PF group mice were reduced, and the kidney damage was also alleviated accordingly. Moreover, the IRAK1/inhibitor of the nuclear factor-κB kinase (IKK)/NF-κB inhibitor (IκB)/NF-κB pathways was found to be involved in the anti-inflammation effect of PF in the kidney and spleen. In conclusion, it is thought that PF may have the potential to be used as a therapeutic agent to reduce the inflammatory activity of SLE. Inhibition of the IRAK1-NF-κB pathway may help formulate novel therapeutic tactics for SLE.

Scorpion venom polypeptide governs alveolar macrophage M1/M2 polarization to alleviate pulmonary fibrosis.

The onset of connective tissue disease-related interstitial lung disease (CTD-ILD) is generally insidious and progressive, with pulmonary fibrosis in the middle stage, and eventually respiratory failure and death. This study aimed to explore the role of scorpion venom polypeptide (SVP), the primary active constituent of the entire scorpion, in alveolar macrophages and pulmonary fibrosis. Pulmonary fibrosis mouse models were established, and then SVP and JAK inhibitor (tofacitinib) was used to treat models. Alveolar macrophages were isolated and the impacts of SVP on M1/M2 polarization and the JAK/STAT6 pathway in vitro were assessed. H&E and Masson staining revealed that SVP and tofacitinib treatment alleviated lung damage and fibrosis. They also hindered the M2-polarization of macrophages in lung tissue and declined cytokine levels associated with M2 polarization (IL-4, IL-13) and fibrosis drivers (TGF-ß, VEGF) in mice. Consistent with the trend presented by tofacitinib treatment, SVP suppressed the phosphorylation of proteins in the JAK/STAT6 pathway. In addition, the in vitro treatment of SVP on the isolated macrophages represented consistent results with in vivo experiments. The findings of the present study indicated that SVP suppressed the JAK/STAT6 signaling pathway, hindered alveolar macrophage M2-type polarization, and possessed the potential to ameliorate pulmonary fibrosis.

Investigation of the mechanism of action of Shengxuexiaoban Capsules against primary immune thrombocytopenia using network pharmacology and experimental validation.

BACKGROUND: Shengxuexiaoban Capsules (SC) is a classical prescription in traditional Chinese medicine (TCM) and has been clinically adopted in the treatment of primary immune thrombocytopenia (ITP) in China. However, the underlying mechanisms of the actions of SC on ITP remain clear. METHODS: A network pharmacology approach was adopted to investigate the underlying molecular mechanism of SC in treating ITP, and the effects of SC on the proliferation, differentiation, and apoptosis of megakaryocyte (MK) and on the ITP animal model were investigated. RESULTS: Network pharmacology analysis found 128 active compounds and 268 targets of these compounds in SC, as well as 221 ITP-related targets. The topological analysis found a central network containing 82 genes, which were significantly associated with the regulation of transcription, cell proliferation, apoptosis processes, the PI3K-AKT signaling pathway, the MAPK signaling pathway, and the ERK1 and ERK2 cascades. It showed that SC increased the proliferation and differentiation of MK, but had no significant impact on MK apoptosis in vivo. The addition of SC increased the gene expression of several potential targets, including STAT3, KDR, CASP3, and TGFB1. In addition, SC administration elevated the protein expression of p-AKT and inhibit the protein expression of p-ERK, but has no impact on the protein expression of p-P38. Moreover, SC could improve haemogram parameters, coagulation indicators, and the proliferation and differentiation of MK in the ITP animal model. CONCLUSIONS: The present study systematically elucidated the underlying mechanisms of SC against ITP and provided an efficient strategy to discover the pharmacological mechanism of TCM. It may strengthen the understanding of SC and facilitate more application of this formula in the treatment of ITP.

Hypertrophic pachymeningitis with co-occurrence of anti-neutrophil cytoplasmic antibodies and immunoglobulin G4 seropositivity: Case report and literature review.

We report an extremely rare case of hypertrophic pachymeningitis in which a 71-year-old man presented with an intractable recurrent headache for >1 year. During this period, he became positive for immunoglobulin G4 and proteinase 3-antineutrophil cytoplasmic antibodies. Contrast-enhanced magnetic resonance imaging showed characteristic diffuse thickening of the dura. Symptoms were improved by intravenous methylprednisolone (500 mg per day for 5 days) and cyclophosphamide pulse therapy during corticosteroid withdrawal; he remained symptom-free during 1-year follow-up. This case suggests that this disease can be treated by corticosteroids combined with immunosuppressive agents.

Eltrombopag improves refractory thrombocytopenia in patients with Sjögren's syndrome.

INTRODUCTION: Eltrombopag, a kind of thrombopoietin (TPO) receptor agonist, plays the role on the megakaryocyte to activate the platelet production and rapidly increase the number of circulating platelets in patients with primary immune thrombocytopenia (ITP). Eltrombopag provides an opportunity for rapid tapering and/or cessation of corticosteroid therapy. However, it is not clear about the platelet response to Eltrombopag in ITP associated with Sjögren's Syndrome(SS). METHODS: A retrospective research was conducted on the clinical course of three patients, each with ITP secondary to SS, and initially received therapy of corticosteroids or other immunomodulatory. They took this drug for bleeding diseases. Referring to the description, Eltrombopag was prescribed and adjusted with an initial dose of 25 mg daily, then weekly, then monthly according to the monitoring of platelet counts. RESULTS: All patients maintained a satisfactory level of platelet counts (>100,000/mm3 for >2 years) following corticosteroid withdrawal. Meanwhile, Eltrombopag was well-tolerated, and there were no adverse effects, such as thrombotic events. CONCLUSIONS: Eltrombopag is effective and safe for patients with ITP associated with SS during corticosteroid withdrawal. Thus it may be a crucial therapeutic strategy for reducing corticosteroid-related side effects in SS patients with ITP.

Scorpion Venom Polypeptide Inhibits Pulmonary Epithelial-Mesenchymal Transition in Systemic Sclerosis-Interstitial Lung Disease Model Mice by Intervening TGF-ß1/Smad Signaling Pathway.

Objective: Interstitial lung disease (ILD) is an important complication of systemic sclerosis (SSc). The aim of this study was to investigate the effect and possible mechanism of polypeptide extract of scorpion venom (PESV) on SSc-ILD. Methods: C57/BL6 mice were injected with bleomycin to establish a SSc-ILD model. Different concentrations of PESV solution were administered to SSc-ILD mice, and dexamethasone was used as a positive control. H&E staining and Masson staining were used to observe the pathological changes. The TGF-ß1 expression level was detected by immunohistochemistry. The expression of epithelial-mesenchymal transition (EMT)-related proteins was detected by Western blot, and the expression of TGF-ß1/Smad pathway-related proteins was also detected. The content of inflammatory cytokines in serum and BALF was determined by ELISA. Results: Pathological analysis showed that PESV could alleviate SSc-ILD-induced pulmonary inflammation and fibrosis. Compared with the model group, the content of inflammatory cytokines IL-6 and TNF-α significantly decreased after PESV treatment. PESV could increase the expression of epithelial marker (E-cadherin) and reduce the expression of interstitial markers (collagen I, vimentin, N-cadherin, and a-SMA). In addition, PESV could reduce the expression level of TGF-ß1/Smad pathway-related protein. Conclusion: PESV can attenuate SSc-ILD by regulating EMT, and the effect was linked to the TGF-ß1/Smad signaling pathway, which indicated that PESV may serve as a candidate drug for SSc-ILD.

Successful treatment of refractory cystitis associated with systemic lupus erythematosus with Belimumab.

Lupus cystitis (LC) is a rare manifestation of SLE, the diagnosis of LC is challenging, especially in the absence of other systemic manifestations or the obvious disease activity index of SLE, further cystoscopy and bladder biopsy are crucial. The symptoms of cystitis could be controlled with high-dose GC, but in the process of GC reduction, the condition repeated. Here we report one case of refractory LC treated with Belimumab. The case is a 45-year-old female patient who had SLE and presented with urinary urgency, frequency and pain for more than 3 years. Laboratory assays revealed high ANA, reduced complement 3 level, proteinuria, significantly elevated Leukocyte esterase and leukocyte in urine, with the negative urinary culture of bacteria and mycoplasma, meanwhile, the cystoscopy and bladder biopsy showed interstitial inflammation. Confirming the diagnosis of refractory SLE-LC, recommended-dose Belimumab (10 mg/kg fortnightly for 3 times, monthly for 6 times) was administered, resulting in the normalization of urinary activity and significant reductions in leukocyte counts and protein levels of urine. No lupus cystitis relapse or SLE activity occurred during 10 months of follow-up. Our case confirmed the efficacy and good follow-up outcomes of Belimumab treatment for refractory SLE-LC.

Efficacy and safety of leflunomide in psoriatic arthritis treatment: A single-arm meta-analysis.

AIM: To systematically assess the clinical efficacy and safety of leflunomide for treating psoriatic arthritis (PsA). METHODS: PubMed, Embase, and Cochrane Library databases were searched until 1 January, 2018, to include relevant studies in this single-arm meta-analysis. Psoriasis improvement was evaluated using Psoriasis Area and Severity Index (PASI) scores. The primary outcome in patients was assessed using the Psoriatic Arthritis Response Criteria (PsARC). Other effectiveness evaluations included those of adverse events, quality of life including functional status (Health Assessment Questionnaire [HAQ] total score), and a quality-of-life instrument for dermatologic diseases (Dermatology Life Quality Index [DLQI] total score). RESULTS: A total of 6 studies were included. After leflunomide treatment, 48% of the patients experienced a reduction of ≥50% based on PASI scores (95% confidence interval [CI]: 0.22-0.73). PASI 75 improvement was observed among 25% of the patients (95% CI: 0.11-0.38). The primary effectiveness analysis revealed that 15% of the patients (95% CI: 0.07-0.26) discontinued leflunomide treatment. Further, 77% of the patients (95% CI: 0.59-0.92) achieved a PsARC response. Adverse events occurred in 38% of the patients (95% CI: 0.04-0.71). The mean ± SD percentage PASI improvement was -4.88 (95% CI: -8.92, -0.85). Moreover, DLQI and HAQ were -2.02 (95% CI: -3.01, -1.03) and -0.19 (95% CI: -0.29, -0.09), respectively. CONCLUSIONS: Leflunomide is an effective and well-tolerated treatment for PsA, and would be a safe and convenient option.

Curcumin alleviates rheumatoid arthritis-induced inflammation and synovial hyperplasia by targeting mTOR pathway in rats.

PURPOSE: Rheumatoid arthritis (RA) is a chronic, progressive autoimmune disease characterized by aggressive and symmetric polyarthritis. Mammalian target of rapamycin (mTOR) was reported to be a new target for RA therapy and its inhibitor rapamycin can significantly reduce the invasive force of fibroblast-like synoviocytes. Here, we determined the effect of curcumin to alleviate inflammation and synovial hyperplasia for the therapy of RA. MATERIALS AND METHODS: Collagen-induced arthritis (CIA) was developed in Wistar rats and used as a model resembling RA in humans. Rats were treated with curcumin (200 mg/kg) and the mTOR inhibitor rapamycin (2.5 mg/kg) daily for 3 weeks. Effects of the treatment on local joint, peripheral blood, and synovial hyperplasia in the pathogenesis of CIA were analyzed. RESULTS: Curcumin and rapamycin significantly inhibited the redness and swelling of ankles and joints in RA rats. Curcumin inhibited the CIA-induced mTOR pathway and the RA-induced infiltration of inflammatory cells into the synovium. Curcumin and rapamycin treatment inhibited the increased levels of proinflammatory cytokines including IL-1ß, TNF-α, MMP-1, and MMP-3 in CIA rats. CONCLUSION: Our findings show that curcumin alleviates CIA-induced inflammation, synovial hyperplasia, and the other main features involved in the pathogenesis of CIA via the mTOR pathway. These results provide evidence for the anti-arthritic properties of curcumin and corroborate its potential use for the treatment of RA.

Neurogenic pulmonary edema caused by spontaneous cerebellar hemorrhage: A fatal case report.

BACKGROUND: Neurogenic pulmonary edema (NPE) is a clinical syndrome characterized by an acute increase of pulmonary interstitial and alveolar fluid. It could result from a significant central nervous system (CNS) insult such as intracranial hemorrhage. However, NPE as a major presenting manifestation of cerebellar hemorrhage was seldom reported. CASE DESCRIPTION: We introduce a rare case of a 34-year-old woman suffering from a fulminant NPE in parallel with a spontaneous cerebellar hemorrhage. Although appropriate supportive measures were taken in the neuroscience care unit, the patient failed to survive hypoxemia within 28 h after hospital admission. CONCLUSION: Pathological lesions of the cerebellum may initiate a cascade of reactions including massive sympathetic discharge and catecholamine storm, leading to a dysfunction of both cardiovascular and respiratory systems. By far, no effective therapeutic strategies have been utilized to treat such a situation. Our present report would shed light on the underlying mechanism of NPE.

Biomarkers for renal transplantation: where are we?

Although surgical techniques, post-transplant care medicine, and immunosuppressants have been greatly improved, permanent acceptance of renal allograft remains a clinical challenge owing to the appearance of various influencing factors. To predict graft dysfunction, development of noninvasive biomarkers is becoming a highlighted research topic in the field of renal transplantation, which provides a possibility for physicians to give preemptive rescue treatment. From the viewpoint of diagnostic techniques, repetitive sampling is prerequisite to identify applicable biomarkers in the clinic. Early biomarkers can be used to dynamically monitor renal graft status and accurately predict transplant outcome independent of various confounders. This review highlights recent studies on the predictive value of biomarkers and methods to quantify biomarkers for monitoring kidney transplant. It is important to analyze and compare different biomarkers for living, and nonliving donors. Analysis of identified clinically relevant biomarkers will advance our understanding of distinct molecular and cellular mechanisms of transplantation and provide insight into developing novel potential approaches to induce transplant tolerance.

[Treatment of severe active systemic lupus erythematosus by PMC therapy combined langchuang fuzheng jiedu capsule: a clinical observation].

OBJECTIVE: To evaluate the efficacy and safety of PMC therapy (Prednisone, Methotrexate, Chloroquine) combined Langchuang Fuzheng Jiedu Capsule (LFJC), thus choosing a better therapy of integrative medicine for SLE in the period of glucocorticoid use. METHODS: Sixty active SLE patients were randomly assigned to two groups, the control group and the treatment group. Those in the control group received PMC therapy (As for Prednisone, it was given at the daily dose of 1 mg/kg till 2 weeks after the condition being stable or after 8 weeks of treatment. Then the dose was reduced by 10% every two weeks. When the dose was reduced to 0.5 mg/kg daily, it was reduced by 2.5 mg per two weeks. When the dose was reduced to 15 mg daily, the dose was reduced to 2.5 mg per four weeks. As for Methotrexate, 10 mg each time, once a week. As for Chloroquine, 100 mg each time, twice daily), while those in the treatment group received PMC therapy (the same way as that for the control group) combined with LFJC (consisting of Astragalus membranaceus 50 g, Angelica sinensis 20 g, Ligusticum Chuanxiong 20 g, prepared Rehmannia Rhizome 30 g, Herba Serissae 30 g, Centella 30 g, centipede 4 g, scorpions 10 g, nidus versace 12 g, et al., 0.5 g per pill, containing 5.7 g crude drug. When the hormone was given at a large dose, LFJC was administered at 12 pills each time, three times daily). When the hormone was given at a middle dose, LFJC was administered at 8 pills each time, three times daily. When the hormone was given at a small dose, LFJC was administered at 6 pills each time, three times daily. The treatment course was six months. The improvement of symptoms and signs between before and after treatment, SLE disease activity index (SLEDAI), efficacy of Chinese medical syndrome, UPro quantitation, erythrocyte sedimentation rate (ESR), complement 3 (C3), C-reactive protein (CRP), the reduction and withdrawal of hormones, and infection of the respiratory tract were observed. RESULTS: The difference in post-SLEDAI was obviously larger in the treatment group than in the control group (P < 0.05). The fatigue severity scale (FSS) was less after treatment than before treatment in the treatment group, showing statistical difference when compared with that of the control group (P < 0.05). The total effective rate was 93.33% in the treatment group, showing statistical difference when compared with that of the control group (86.66%; chi2 = 6.736, P < 0.05). The ESR decreased after treatment in the treatment group, showing statistical difference when compared with that of the control group (P < 0.01). C3 increased after treatment in the treatment group, showing statistical difference when compared with that of the control group (P < 0.05). The hormone was reduced to (13.70 +/- 5.42) mg/d by the end of the therapeutic course in the treatment group, obviously less than that of the control group [(17.63 +/- 7.80) mg/d, P < 0.05). Seven patients suffered from secondary infection of the respiratory tract infection in the treatment group (5 from upper respiratory tract infection and 2 from lower respiratory tract infection), obviously less than those of the control group (25 from upper respiratory tract infection and 10 from lower respiratory tract infection) (P < 0.05). CONCLUSIONS: PMC combined LFJC was a better treatment program for severe active SLE (SLEDAI > or = 15). It was more safe and effective when compared with using Western medicine alone. It could enhance the efficacy of hormones and help reduction/withdrawal of hormones.

[Molecular cloning of a novel retroviral NP9 gene of human endogenous retrovirus and its viral protein expression].

OBJECTIVE: To investigate the relationship between retroviruses and autoimmune diseases, to clone the novel retroviral NP9 gene from human endogenous retrovirus (HERV), and to construct its expression vector. METHODS: The viral NP9 gene was amplified and cloned by RT-PCR and T-A clone techniques, and its sequence was determined with Perkin-Elmer 377 DNA Sequencer. The amplified viral NP9 gene was subcloned into the prokaryotic express vector pQE30. The recombinant plasmids were identified by restriction endonuclease digestion and sequencing. The recombinant pQE30-NP9 protein was expressed in M15 host cells under the IPTG induction and showed with SDS-PAGE,and the corresponding NP9 viral protein was identified with Western blot analysis. RESULT: A specific band of 250 bp was amplified using RT-PCR from total RNA of peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE) and confirmed as the NP9 gene via T-A clone and DNA sequencing analyses. SDS-PAGE profile showed a clear protein band with a relative molecular weight 9 kD in the IPTG-induced samples, which was confirmed as viral NP9 protein by Western blot analysis. CONCLUSION: The NP9 gene has been successfully isolated and cloned from PBMCs of SLE patients and the corresponding NP9 viral protein expressed in prokaryotic expression vector.

Expression of a novel human retroviral NP9 gene and potential roles of its protein in systemic lupus erythematosus patients.

OBJECTIVE: To investigate the expression of a novel retroviral (NP9) gene transcripts and the possible role of its protein in systemic lupus erythematosus (SLE) patients. METHODS: The retroviral NP9 gene in SLE patients was isolated and cloned using RT-PCR and TA cloning techniques, and it was analyzed by sequencing. The expression of the NP9 genes in 40 patients with SLE and 48 normal controls using RT-PCR was detected. NCBI BLAST and DNASIS 3.1 software were used to analyze the features of protein of NP9 gene. RESULTS: The positive ratio (77.5%) of the mRNA expression of the retroviral NP9 gene in SLE patients is significantly higher than that (8.3%) in normal subjects (P<0.01). The recombinant NP9 protein comprises 74 AA with pI 9.59. Amino acid sequence analysis indicates that the retroviral NP9 protein shares higher homologies with several human proteins with important biological functions. CONCLUSION: SLE patients possess specific novel retroviral NP9 transcripts. The expression of the retroviral NP9 gene may involve in the genesis or development of SLE.