Current extraction methods often extract DNA and RNA separately, and few methods are capable of co-extracting DNA and RNA from sputum. We established a nucleic acid co-extraction method from sputum based on magnetic beads and optimized the method by evaluating influencing factors, such as the guanidinium thiocyanate (GTC) and dithiothreitol (DTT) concentrations, magnetic bead amount, incubation temperature, lysis buffer pH and RNA carrier type. The feasibility of the simultaneous nucleic acid co-extraction method was evaluated by amplifying DNA and RNA viruses from a single clinical specimen with a multiplex RT-qPCR method. Both DNA and RNA were most efficiently extracted when the GTC and DTT concentrations were 2.0 M and 80 mM, respectively, 20 μl magnetic beads were added, the incubation temperature was 80 °C, the pH was 8 or 9, and RNA carrier A was used. Therefore, we established a simple method to extract nucleic acids from two important respiratory viruses compared with other commercial kits. This magnetic beads-based co-extraction method for sputum followed by a multiplex RT-qPCR can rapidly and precisely detect DNA and RNA viruses from a single clinical specimen and has many advantages, such as decreased time, low cost, and a lack of harmful chemicals.

Jieduquyuziyin prescription (JP) has been used to treat systemic lupus erythematosus (SLE). Although the effectiveness of JP in the treatment of SLE has been clinically proven, the underlying mechanisms have yet to be completely understood. We observed the therapeutic actions of JP in MRL/lpr mice and their bone marrow-derived macrophages (BMDMs) and the potential mechanism of their inhibition of inflammatory activity. To estimate the effect of JP on suppressing inflammatory activity, BMDMs of MRL/lpr and MRL/MP mice were treated with JP-treated serum, and MRL/lpr mice were treated by JP for 8 weeks. Among them, JP and its treated serum were subjected to quality control, and BMDMs were separated and identified. The results showed that in the JP group of BMDMs stimulated by Lipopolysaccharide (LPS) in MRL/lpr mice, the secretion of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) reduced, and the expressions of Interleukin-1 receptor-associated kinase 1 (IRAK1) and its downstream nuclear factor κB (NF-κB) pathway decreased. Meanwhile, the alleviation of renal pathological damage, the decrease of urinary protein and serum anti-dsDNA contents, the inhibition of TNF-α level, and then the suppression of the IRAK1-NF-κB inflammatory signaling in the spleen and kidney, confirmed that the therapeutic effect of JP. These results demonstrated that JP could inhibit the inflammatory activity of MRL/lpr mice and their BMDMs by suppressing the activation of IRAK1-NF-κB signaling and was supposed to be a good choice for the treatment of SLE.

Objective: Hydroxychloroquine (HCQ) is an antimalarial drug employed in the treatment of systemic lupus erythematosus (SLE). Prior studies reported inconsistent results regarding the association between HCQ use during pregnancy and adverse pregnancy outcomes. This study aimed to evaluate the impact of HCQ on pregnancy-related outcomes in women with SLE. Methods: We conducted a systematic search for studies associating pregnancy outcomes with HCQ use in PubMed, EMBASE, Cochrane Library, Web of Science, CNKI, Wanfang database and VIP from inception to 22 September 2022. Random or fixed effect models were used to estimate the pooled effect based on I ² measurement of heterogeneity. Results: Twenty-one studies were included, encompassing 929 and 1031 patients in HCQ and non-HCQ groups, respectively. We found that HCQ use was significantly associated with reduced risks of Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) scores (second trimester: mean difference (MD) −1.80, 95% CI −2.46 to –1.13; third trimester: MD −2.30, 95% CI −3.31 to –1.29), flare (OR 0.57, 95% CI 0.33 to 0.97), preterm birth (OR 0.57, 95% CI 0.46 to 0.72), intrauterine growth retardation (IUGR) (OR 0.48, 95% CI 0.31 to 0.72), gestational hypertension (OR 0.19, 95% CI 0.08 to 0.42), pre-eclampsia (OR 0.46, 95% CI 0.29 to 0.72). In contrast, a positive correlation was observed between full-term birth and HCQ use (OR 2.01, 95% CI 1.52 to 2.65). However, the result for disease flare exhibited high heterogeneity (p=0.01, I ² =59%). In addition, publication bias was detected in the meta-analysis of full-term birth using the Egger’s test. Conclusions: This meta-analysis offers a comprehensive assessment of the relationship between disease activity, pregnancy-related outcomes and HCQ use, providing supportive evidence for the therapeutic effectiveness of HCQ in pregnant women with SLE. PROSPERO registration number: CRD42022374468.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease, and Jieduquyuziyin prescription (JP) is a traditional Chinese medicine (TCM) formula that has been testified to be effective for SLE treatment as an approved hospital prescription for many years in China. However, its mechanism of action in the treatment of this disease is largely unknown. The purpose of this study was to determine whether JP-treated rat serum can inhibit the activation of peritoneal macrophages in MRL/lpr mice by downregulating the IRAK1 signaling pathway, thereby achieving the effect of improving SLE. The JP-treated rat serum was prepared, and the peritoneal macrophages of MRL/lpr lupus mice were isolated in vitro, and the effect of JP on cell viability was detected by the CCK8 method. After LPS induction and shRNA lentiviral transfection, the effect of JP on the expression of IRAK1 in cells was detected by immunofluorescence staining. The content of TNF-α and IL-6 in the cell supernatant was determined by ELISA. The expression of IRAK1, NF-κB, TNF-α, and IL-6 mRNA was detected by RT-PCR, and the expression levels of IRAK1, p-IRAK1, TRAF6, IKBα, p-IKBα, IKK + IKK, NF-κB, and p-NF-κB proteins was detected by western blot method. We investigated the role of JP in peritoneal macrophages of the MRL/lpr mouse and identified the possible mechanisms of action. The results showed that JP could reduce the phosphorylation of IRAK1 and its downstream proteins induced by LPS and inhibit the expression of inflammatory cytokines, including TNF-α and IL-6. In addition, after the transfection of cells with shRNA lentiviral, the results of JP tended to be consistent. In conclusion, JP may inhibit the activation of peritoneal macrophages in MRL/lpr mice by downregulating the IRAK1-NF-κB signaling pathway, and IRAK1 may be a potential target for JP treatment of SLE.