Development and application of a universal extraction-free reagent based on an algal glycolipid.

In this study, we independently developed a universal nasopharyngeal swab extraction-free reagent based on a trehalose lipid for the rapid detection of pathogen nucleic acids in respiratory infectious diseases. By comparing the isothermal amplification results of a 2019-nCoV pseudovirus solution treated with different components of the extraction-free reagent, we determined the optimal composition of the extraction-free reagent to be a mixed solution of 10 mmol L-1 tris-HCl containing 0.05 mmol L-1 EDTA (TE solution), 5% glycine betaine, 0.5% Triton X-100, and 1.5% trehalose lipid. The results showed that the extraction-free reagent could cleave DNA viruses, RNA viruses, and bacteria to release nucleic acids and did not affect the subsequent nucleic acid amplification. Its efficiency was consistent with that of magnetic bead extraction. Real-time fluorescence quantitative PCR was used to analyze the stability and repeatability of the detection results of the samples treated with the extraction-free reagent and the sensitivity of the extraction-free reagent. The results showed that the extraction-free kit could stably store the pathogen nucleic acid for at least 24 hours, the detection repeatability was satisfactory, and there was no incompatibility with the detection limits of various manufacturers' nucleic acid detection reagents. In conclusion, the established nucleic acid extraction-free method can effectively lyse respiratory infectious disease pathogens to release nucleic acids (DNA and RNA) at room temperature and can directly amplify nucleic acids without extraction steps. This method takes a short time and has high efficiency. The released nucleic acid met the requirements of molecular biological detection methods such as real-time fluorescence quantitative PCR (qPCR), reverse transcription-polymerase chain reaction (RT-PCR), and isothermal nucleic acid amplification (INAA).

Overexpressed RAD51 promoted osteogenic differentiation by activating IGF1R/PI3K/AKT pathway in osteoblasts.

BACKGROUND: Osteoporosis (OP) is a skeleton disease induced by imbalance between osteoblast and osteoclast. Osteogenic differentiation of osteoblasts is of great importance, and the regulatory mechanisms are urgent to be studied. METHODS: Differentially expressed genes were screened from microarray profile related to OP patients. The dexamethasone (Dex) was used to induce osteogenic differentiation of MC3T3-E1 cells. MC3T3-E1 cells were exposed to microgravity environment to mimic OP model cells. Alizarin Red staining and alkaline phosphatase (ALP) staining were used to evaluate the role of RAD51 in osteogenic differentiation of OP model cells. Furthermore, qRT-PCR and western blot were applied to determine expression levels of genes and proteins. RESULTS: RAD51 expression was suppressed in OP patients and model cells. Alizarin Red staining and ALP staining intensity, the expression of osteogenesis-related proteins including runt-related transcription factor 2 (Runx2), osteocalcin (OCN), and collagen type I alpha1 (COL1A1) were increased by over-expressed RAD51. Furthermore, RAD51 related genes were enriched in IGF1 pathway, and up-regulated RAD51 activated IGF1 pathway. The effects of oe-RAD51 on osteogenic differentiation and IGF1 pathway were attenuated by IGF1R inhibitor BMS754807. CONCLUSIONS: Overexpressed RAD51 promoted osteogenic differentiation by activating IGF1R/PI3K/AKT signaling pathway in OP. RAD51 could be a potential therapeutic marker for OP.

The pattern and trends of disease burden due to low bone mineral density from 1990 to 2019 in China: findings from the Global Burden of Disease Study 2019.

Osteoporosis is becoming increasing important health problem in China. This study shows that the disease burden of low bone mineral density (BMD) in China is large and will remain increasing with the growth of aging population. In addition, male low BMD should not be ignored. Although burden of low BMD is partially representative of the real burden of osteoporosis, the information provided in our study could be used to better inform targeted public health prevention and management programs for osteopososis. PURPOSE: We aim to investigate the pattern and trends of disease burden due to low BMD by gender, year, and age in China from 1990 to 2019. METHODS: Data on summary exposure value (SEV) and disability-adjusted life years (DALYs) due to low BMD was obtained from the Global Burden of Disease Study 2019, and analyzed by gender, age, and years. Average annual percent change (AAPC) and annual percent change (APC) were calculated to qualify the trends of burden due to low BMD. RESULT: In 2019, the age-standardized SEV was higher in females than that in males (23.04, 95% UI = [17.25-29.83] and 12.50, 95% UI = [7.71-19.25], respectively), while the total number of DALYs was higher in males than females with 1,698,705.92 (95% UI = 1,281,580.79 to 2,076,364.25) and 1,621,569 (95% UI = 1,266,284.89 to 2,016,399.16), respectively. Though SEV exhibited decreasing trends during 1990 to 2019 in both sexes, the absolute number of DALYs due to low BMD increased steadily and almost doubled in 2019 compared to that in 1990. CONCLUSION: The burden of low BMD remains large and continues to increase. Although females are prone to low BMD, the disease burden for males should not be ignored.

Ataxia-televangelist mutated (ATM)/ ATR serine/threonine kinase (ATR)-mediated RAD51 recombinase (RAD51) promotes osteogenic differentiation and inhibits osteoclastogenesis in osteoporosis.

Osteoporosis is a metabolic bone disease that significantly affects the quality of life and can even lead to death. In this study, we aimed to investigate the role of RAD51 recombinase (RAD51) in osteoblast and osteoclast differentiation. We analyzed differentially expressed genes using microarray analysis. The osteogenic differentiation capability was analyzed by alkaline phosphatase (ALP) staining and alizarin red staining assays. Osteogenesis and osteoclast related genes expression was detected using quantitative real-time PCR (qPCR) and Western blotting. The phosphorylation of Ataxia-telangiectasia mutated (ATM) and ATR serine/threonine kinase (ATR) was tested using Western blotting. The effect of RAD51 on osteoporosis was also explored in vivo. The results showed that RAD51 was downregulated in osteoporosis, but upregulated in differentiated osteoblasts. Overexpression of RAD51 enhanced the differentiation of osteoblasts and suppressed the formation of osteoclasts. Furthermore, p-ATM and p-ATR levels were upregulated in osteoblasts and downregulated in osteoclasts. RAD51 expression was reduced by the ATM/ATR pathway inhibitor AZ20. AZ20 treatment inhibited osteoblastogenesis and promoted osteoclastogenesis, whereas RAD51 reversed the effects induced by AZ20. Moreover, RAD51 improved bone microarchitecture in vivo. Taken together, ATM/ATR signaling-mediated RAD51 promoted osteogenic differentiation and suppressed osteoclastogenesis. These findings reveal a critical role for RAD51 in osteoporosis.

Identification of the urine and serum metabolomics signature of gout.

OBJECTIVE: Gout is the most common inflammatory arthritis and the worldwide incidence is increasing. By revealing the metabolic alterations in serum and urine of gout patients, the first aim of our study was to discover novel molecular biomarkers allowing for early diagnosis. We also aimed to investigate the underlying pathogenic pathways. METHODS: Serum and urine samples from gout patients (n = 30) and age-matched healthy controls (n = 30) were analysed by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) to screen the differential metabolites and construct a diagnostic model. Next, the model was verified and optimized in the second validation cohort (n = 100). The pathways were illustrated to understand the underlying pathogenesis of gout. RESULTS: In general, serum metabolomics demonstrated a clearer distinction than urine metabolomics. In the discovery cohort, 40 differential serum metabolites were identified that could distinguish gout patients from healthy controls. Among them, eight serum metabolites were verified in the validation cohort. Through regression analysis, the final model consisted of three serum metabolites-pyroglutamic acid, 2-methylbutyryl carnitine and Phe-Phe-that presented optimal diagnostic power. The three proposed metabolites produced an area under the curve of 0.956 (95% CI 0.911, 1.000). Additionally, the proposed metabolic pathways were primarily involved in purine metabolism, branched-chain amino acids (BCAAs) metabolism, the tricarboxylic acid cycle, synthesis and degradation of ketone bodies, bile secretion and arachidonic acid metabolism. CONCLUSION: The metabolomics signatures could serve as an efficient tool for early diagnosis and provide novel insights into the pathogenesis of gout.

Age-Specific Imbalance of Circulating Tfh Cell Subsets and Its Association With Gout-Targeted Kidney Impairment.

Objective: Gout is a chronic disease characterized by the deposition of monosodium urate (MSU) crystals in tissue. Study with a focus on adaptive immune response remains to be understood although innate immune response has been reported extensively in gout etiology. Our study attempted to investigate the association of gout-related immune cell imbalance with clinical features and comorbidity with renal impairment and the implicated pathogenesis via the assessment of T and B cell subsets in different activity phases or with immune effects combined with the analyses of clinical parameters. Methods: Fifty-eight gout patients and 56 age- and sex-matched healthy individuals were enrolled. To learn the roles of circulating T cells, a lymphocyte profile incorporating 32 T cell subsets was tested from isolated freshly peripheral blood monocyte cells (PBMCs) with multiple-color flow cytometry. Furthermore, the collected clinical features of participants were used to analyze the characteristics of these differential cell subsets. Stratified on the basis of the level of creatinine (Cr, enzymatic method), all patients were categorized into Crlow (Cr ≤ 116 µmol/L) and Crhi (Cr > 116 µmol/L) groups to exploit whether these gout-associated T cell subsets were functional in gout-targeted kidney dysfunction. The differentiation of B cells was investigated in gout patients. Results: Our results show that CD 4+ T cells, Th2 cells, and Tc2 cells were upregulated, whereas Tc17 cells were downregulated. Tfh cells skewed toward the polarization of Tfh2 cells. Specifically, Tfh2 cells increased, but Tfh1 cells decreased, accompanied with aging for gout patients, suggesting that age might trigger the skewing of Tfh1/Tfh2 cell subsets to influence gout development. Moreover, Tfh2 cells were connected to renal dysfunction as well. No alterations of B cell subsets were observed in patients when compared to controls. Conclusions: Our data demonstrate age-specific dysfunctions of Tfh1/2 cells in gout occurrence, and Tfh2 cell upregulation is associated with gout-targeted renal dysfunction. However, Tfh2 cells may function in auto-inflammatory gout independent of helping B differentiation, and an in-depth study remains to be conducted.

Using Mobile Apps for Health Management: A New Health Care Mode in China.

BACKGROUND: China has a large population; however, medical resources are unevenly distributed and extremely limited, and more medical services are needed. With the development and ever-increasing popularity of mobile internet communication, China has created a mode of mobile health (mHealth) care to resolve this problem. OBJECTIVE: The aim of this study was (1) to describe the problems associated with China's medical care practice, (2) explore the need for and the feasibility of internet-based medical care in China, and (3) analyze the functionality of and services offered by internet-based health care platforms for the management of chronic diseases. METHODS: Data search was performed by searching national websites, the popular search engine Baidu, the App Store, and websites of internet medical care institutions, using search terms like "mobile health," "Internet health," "mobile medical," "Internet medical," "digital medical," "digital health," and "online doctor." A total of 6 mobile apps and websites with the biggest enrollment targeting doctors and end users with chronic diseases in China were selected. RESULTS: We recognized the limitations of medical and health care providers and unequal distribution of medical resources in China. An mHealth care platform is a novel and efficient way for doctors and patients to follow up and manage chronic diseases. Services offered by these platforms include reservation and payment, medical consultation, medical education assessment, pharmaceutical and medical instruments sales, electronic medical records, and chronic disease management. China's health policies are now strongly promoting the implementation of mHealth solutions, particularly in response to the increasing burden of chronic diseases and aging in the population. CONCLUSIONS: China's internet-based medical and health care mode can benefit the populace by providing people with high-quality medical resources. This can help other countries and regions with high population density and unevenly distributed medical resources manage their health care concerns.

Serum from patients with ankylosing spondylitis can increase PPARD, fra-1, MMP7, OPG and RANKL expression in MG63 cells.

OBJECTIVES: To explore the effects of serum from patients with ankylosing spondylitis on the canonical Wnt/ß-catenin pathway and to assess whether the serum has an osteogenic effect in MG63 cells. METHODS: MG63 cells were cultured with serum from 45 ankylosing spondylitis patients, 30 healthy controls, or 45 rheumatoid arthritis patients. The relative PPARD, fra-1, MMP7, OPG and RANKL mRNA levels were measured using quantitative real-time polymerase chain reaction. Associations between gene expression and patient demographics and clinical assessments were then analyzed. RESULTS: MG63 cells treated with serum from ankylosing spondylitis patients had higher PPARD, fra-1, MMP7 and OPG gene expression than did cells treated with serum from controls or rheumatoid arthritis patients (all p<0.05). RANKL expression was higher in MG63 cells treated with serum from patients with ankylosing spondylitis or rheumatoid arthritis than in those treated with serum from controls (both p<0.05). The OPG/RANKL ratio was also higher in MG63 cells treated with serum from ankylosing spondylitis patients than in those treated with serum from controls (p<0.05). No associations were found between the expression of the five genes and the patient demographics and clinical assessments (all p>0.05). CONCLUSIONS: Serum from ankylosing spondylitis patients increases PPARD, fra-1, MMP7, OPG and RANKL expression and the OPG/RANKL ratio in MG63 cells; these effects may be due to the stimulatory effect of the serum on the Wnt pathway.

Serum from patients with ankylosing spondylitis can increase PPARD, fra-1, MMP7, OPG and RANKL expression in MG63 cells

OBJECTIVES: To explore the effects of serum from patients with ankylosing spondylitis on the canonical Wnt/β-catenin pathway and to assess whether the serum has an osteogenic effect in MG63 cells. METHODS: MG63 cells were cultured with serum from 45 ankylosing spondylitis patients, 30 healthy controls, or 45 rheumatoid arthritis patients. The relative PPARD, fra-1, MMP7, OPG and RANKL mRNA levels were measured using quantitative real-time polymerase chain reaction. Associations between gene expression and patient demographics and clinical assessments were then analyzed. RESULTS: MG63 cells treated with serum from ankylosing spondylitis patients had higher PPARD, fra-1, MMP7 and OPG gene expression than did cells treated with serum from controls or rheumatoid arthritis patients (all p<0.05). RANKL expression was higher in MG63 cells treated with serum from patients with ankylosing spondylitis or rheumatoid arthritis than in those treated with serum from controls (both p<0.05). The OPG/RANKL ratio was also higher in MG63 cells treated with serum from ankylosing spondylitis patients than in those treated with serum from controls (p<0.05). No associations were found between the expression of the five genes and the patient demographics and clinical assessments (all p>0.05). CONCLUSIONS : Serum from ankylosing spondylitis patients increases PPARD, fra-1, MMP7, OPG and RANKL expression and the OPG/RANKL ratio in MG63 cells; these effects may be due to the stimulatory effect of the serum on the Wnt pathway.

Low prevalence of hepatitis B virus infection in patients with systemic lupus erythematosus in southern China.

To investigate the prevalence of hepatitis B virus (HBV) infection in systemic lupus erythematosus (SLE) patients in southern China, SLE inpatients were retrospectively investigated for their HBV infection rate. Fifteen SLE patients positive for hepatitis B surface antigen (HBsAg) were followed up. Furthermore, serum interferon (IFN)-α levels among SLE patients were detected by ELISA. Results showed estimated HBsAg-positive rate was 10.74% in general population. The HBsAg-positive rate was lower in SLE patients compared with controls (2.33 vs. 9.57%, P < 0.01). Interestingly, 13 out of 15 SLE patients converted from HBsAg positive to HBV surface antibody (HBsAb) positive even under glucocorticoid therapy. In addition, we found significantly increased IFN-α levels in SLE patients.The prevalence of HBV infection in SLE patients was lower than that in sex- and age-matched non-SLE controls in southern China. The characteristic IFN signatures in SLE may favor the subsequent clearance of HBV.