[Concept and understanding of non-radiographic axial spondyloarthritis].

Since the publication of the classification criteria for axial spondyloarthritis (axSpA) by Assessment of SpondyloArthritis international Society (ASAS), a new disease concept "non-radiographic axial spondyloarthritis (nr-axSpA)" has emerged. Some domestic experts in China have translated it as "radiologically negative" axSpA, but it can easily lead to misunderstandings literally, as not all patients with nr-axSpA are completely free from radiological abnormalities. This article briefly describes the evolution of the concept of axSpA, proposes to translate the term of nr-axSpA directly rather than "radiologically negative" axSpA, compares the differences and similarities between nr-axSpA and radiologically negative axSpA, delineates the relationship between nr-axSpA and early ankylosing spondylitis, and points out not all the patients who meet the ASAS classification criteria for nr-axSpA are genuine patients with axSpA, because of the relatively low specificity of the ASAS classification criteria. Five common scenarios easily leading to misdiagnosis of nr-axSpA are exampled to remind domestic colleagues.

[Recommendations for diagnosis and treatment of systemic sclerosis in China].

Systemic sclerosis (SSc) is an autoimmune rheumatic disease that is characterized by skin fibrosis with multi-organ involvement. In China, the standardized diagnosis and treatment for SSc is still lacking. Based on the diagnosis criteria and guidelines from China and abroad, Chinese Rheumatology Association developed the current standardization of diagnosis and treatment for SSc. The purposes of this guideline are to standardize clinical management for SSc in China, to interpret the key evaluation tools for SSc, and to recommend therapeutic principle and strategies.

[Expression of annexin A1 in peripheral blood cells of Naïve rheumatoid arthritis patients and its influencing factors].

Objective: To explore the expression of Annexin a1 (AnxA1) mRNA in peripheral blood mononuclear cells (PBMC) from Naïve rheumatoid arthritis (RA) patients and healthy controls and to investigate its influencing factors. Methods: Thirty Naïve RA patients and 36 healthy controls were recruited from the First Affiliated Hospital of Harbin Medical University between May 2013 and January 2015.All RA patients were fulfilled the classification criteria of ACR in 1987.Clinical characteristics and laboratory indexes were collected.Real time polymerase chain reaction (RT-PCR) was used to measure AnxA1 and Foxp3 mRNA expression respectively.Serum TNF-α, IL-17 and IL-10 levels in each group was determined separately by enzyme linked immunosorbent assay (ELISA). Furthermore, correlations between the expression of AnxA1 mRNA and clinical characteristics, laboratory parameters, Foxp3 mRNA, serum IL-17, TNF-α, IL-10 level in Naïve RA patients were analyzed. Results: The expressions of AnxA1 mRNA in PBMC of Naïve RA patients were lower than that of healthy controls [0.12 (0.05-0.30) versus 1.66 (0.40-2.68), P<0.05]. The Foxp3 mRNA expressions in PBMC of Naïve RA patients were also lower than that of healthy controls [0.77 (0.39-1.89) versus 2.93 (2.65-3.49), P<0.05], while serum TNF-α, IL-17, and IL-10 levels of Naïve RA patients were higher than those of healthy controls (P<0.05). The expression of AnxA1 was positively associated with the expression of Foxp3 mRNA (r(s)=0.513, P<0.05) and negatively associated with the serum IL-17 level (r(s)=-0.381, P<0.05). But there were no correlations between expression of AnxA1 in PBMCs from Naïve RA patients with the TNF-α, IL-10 level, clinical characteristics (sex, age, disease duration and disease activity) and the laboratory indexes (RF, Anti-CCP antibody, ESR and CRP). The expression of Foxp3 mRNA and the serum IL-17 level was the independent influencing factor. Conclusions: The reduced expression of AnxA1 may associate with the reduced expression of Foxp3 and the increased IL-17 level in Naïve RA patients.It suggested that AnxA1 may play a key role in immune regulation and anti-inflammatory process in the pathogenesis of RA.

[A comparative study of diagnostic efficacy of different diagnostic/ classification criteria in Takayasu arteritis].

OBJECTIVE: This study aimed to analyze and evaluate the diagnostic efficacy of Ishikawa's, the modified Ishikawa's criteria, 1990 American College of Rheumatology (ACR ) classification criteria and the diagnostic model based on Chinese population in Chinese TA patients. METHODS: One hundred and forty-nine patients with Takayasu arteritis and 126 patients with other vascular disorders which involved aorta or its branches were recruited in this study.All the patients were admitted to the Department of rheumatism and Immunology clinic or inpatient department of Zhongshan Hospital affiliated to Fudan University from January 1(st), 2008 to June 31(st), 2015.General characteristics, clinical manifestations, laboratory results and imaging data of all the patients were collected.Sensitivity, specificity, accuracy and area under receiver operating characteristics (ROC) curve of different criteria were analyzed. RESULTS: Sensitivity, specificity, accuracy and area under ROC curve of Chinese diagnostic model were 90.60%, 80.95%, 86.18%, and 85.80%, respectively, while those of Ishikawa criteria were 34.23%, 99.21%, 64.00%, and 66.70%, respectively.These four indicators of the modified Ishikawa criteria were 84.13%, 79.87%, 81.82%, and 82.00%, respectively and that of ACR criteria were 83.89%, 83.33%, 83.64%, and 83.60%, respectively.No significant difference was found between any two of Chinese diagnostic model, the modified Ishikawa criteria and ACR criteria in all the indicators.Sensitivity of Chinese diagnostic model was highest, while specificity of Ishikawa criteria was the highest.Among these four criteria, the diagnostic efficacy of Chinese model was the best and that of Ishikawa criteria was the worst. CONCLUSION: Chinese diagnostic model, which is based on Chinese population and adopts advanced imaging modality, has better diagnostic efficacy.

[Prevalence of Clonorchis sinensis infections in freshwater fish in mainland China: A meta-analysis].

OBJECTIVE: To understand the real prevalence of Clonorchis sinensis infections in the freshwater fish in mainland China, so as to provide insights into clonorchiasis control and detection of freshwater fish. METHODS: All literatures reporting the prevalence of C. sinensis infections in the freshwater fish, the second intermediate host of the parasite, were jointly retrieved in Chinese and English electronic databases from January 1, 2010 to December 31, 2020, including Wanfang Data, CNKI, PubMed, Web of Science, Embase and Cochrane Library. All studies were screened based on inclusion and exclusion criteria, and the quality of all enrolled literatures was evaluated. The pooled prevalence of C. sinensis infections in freshwater fish and its 95% confidence interval (CI) were estimated using the software Stata version 15.0, and subgroup analyses were performed to investigate the region-, season- and sample source-specific pooled prevalence of C. sinensis infections in freshwater fish. In addition, the sensitivity and publication bias of all included studies were analyzed. RESULTS: A total of 40 eligible literatures were included in this study, including 37 Chinese literatures and 3 English literatures, and there were 10 high-quality literatures, 27 moderate-quality literatures and 3 low-quality literatures. A total of 53 species containing 37 959 freshwater fish were reported in these 40 studies, and 73.58% (39/53) of freshwater fish species were identified with C. sinensis infections. Meta-analysis showed 23.5% [95% CI: (0.19, 0.28)] pooled prevalence of C. sinensis infections in freshwater fish in mainland China, and subgroup analyses higher prevalence of C. sinensis infections in freshwater fish in northeastern China [35.7%, 95% CI: (0.22, 0.50)] than in central [25.9%, 95% CI: (0.04, 0.48)] and southern China [20.6%, 95% CI: (0.09, 0.32)], higher prevalence of C. sinensis infections in freshwater fish sampled in spring [44.1%, 95% CI: (0.35, 0.53)] than in autumn [6.7%, 95% CI: (0.05, 0.08)] and summer [3.3%, 95% CI: (-0.01, 0.07)], and higher prevalence of C. sinensis infections in freshwater fish sampled from natural water [25.2%, 95% CI: (0.17, 0.33)] than from retail trades [22.2%, 95% CI: (0.17, 0.28)] and breeding chain [12.3%, 95% CI: (0.03, 0.22)]. However, all included studies had a publication bias with a low sensitivity. CONCLUSIONS: The prevalence of C. sinensis infections is high in freshwater fish in mainland China, and there are still challenges for clonorchiasis control. Reinforcement of health education, diagnostics development and food safety supervision is recommended in future clonorchiasis control programs.

[Endemic situation of schistosomiasis in People's Republic of China from 2010 to 2017].

OBJECTIVE: To understand the epidemic trend of schistosomiasis in China from 2010 to 2017 so as to provide the scientific evidence for schistosomiasis elimination. METHODS: The information of schistosomiasis control nationwide from 2010 to 2017 was collected, including the endemic of population, status of livestock control, and Oncomelania hupensis snail control. Microsoft Excel was applied for datum management and analysis. RESULTS: From 2010 to 2017, the epidemic of schistosomiasis in China dropped significantly. The decreasing amplitude of estimated number of patients nationwide was 88.46%. Seventy-one acute schistosomiasis patients were reported and 12.68% (9/71) of them were imported. The decreasing rate of cultivated cattle was 50.09%, and the accumulative number of schistosome-infected cattle was 17 239, and the average positive rate of stool examinations decreased from 1.04% to 0.000 22%. The area with snails nationwide was 373 596.18 to 363 068.95 hm2, and the new detected area with snails was 46.71 to 1 346.73 hm2. The area with schistosome-infected snails was 171.68 hm2 in 2012 and it was 9.25 hm2 in 2013. In 72 key monitoring points of 7 endemic provinces, there were 17 schistosome positive points of water body in 2010 and 6 points in 2016. There were some high risk-factors related to schistosomiasis transmission including schisto-some-infected cattle, dogs, and field rats, and the field stools, and the pasture in the area with snails in schistosomiasis monitoring points. CONCLUSIONS: The endemic status of schistosomiasis in China has dropped significantly, and the transmission level is very low. However, the infectious source and risk factors in the endemic environments have not be eliminated. Therefore, the infectious source control, health education, snail control, and transmission monitoring should be strengthened, so as to promote the progress of schistosomiasis elimination.

[Endemic status of schistosomiasis in People's Republic of China in 2018].

This report presented the endemic status of schistosomiasis in the People's Republic of China at a national level in 2018, and analyzed the data collected from the national schistosomiasis prevention and control system and 453 national schistosomiasis surveillance sites. Among the 12 provinces (municipality and autonomous region) endemic for schistosomiasis in China, 5 provinces (municipality and autonomous region), including Shanghai, Zhejiang, Fujian, Guangdong and Guangxi, continued to consolidate the achievements of schistosomiasis elimination, Sichuan Province achieved transmission interruption and 6 provinces of Yunnan, Jiangsu, Hubei, Anhui, Jiangxi and Hunan achieved transmission control by the end of 2018. There were 450 endemic counties (cities, districts) covering 260 million people, specifically including 28 456 endemic villages covering 70.059 7 million people at risk of infection. Among the 450 endemic counties (cities, districts), 58.44% (263/450), 27.56% (124/450) and 14.00% (63/450) reached the criteria of elimination, transmission interruption and transmission control, respectively. By the end of 2018, a total of 29 214 advanced schistosomiasis cases were documented in China. In 2018, a total of 11.127 6 million individuals received inquiry examinations and 2.062 9 million were positive; 7.191 4 million individuals received serological tests and 138.5 thousand of them were positive, 532.2 thousand individuals received stool examinations and 8 were positive in China. In 2018, snail survey was performed in 19 821 endemic villages and Oncomehania snails were found in 7 321 villages, accounting for 36.94% of all surveyed villages, with 3 newly detected villages with snails in China. Snail survey covered an area of 590 241.01 hm2 and 168 319.41 hm2 snail habitats were found, including emerging snail habitats of 61.28 hm2; however, no infected snails were identified. In 2018, a total of 646 823 bovines were raised in the schistosomiasis endemic areas of China, and 225 258 received serological examinations, with 2 638 positives detected, while 164 803 bovines received stool examinations, with 2 positives identified. In 2018, there were 90 388 patients with schistosomiasis receiving praziquantel chemotherapy, and expanded chemotherapy was given to 1 490 594 person-times; there were two bovines with schistosomiasis receiving praziquantel chemotherapy, and expanded chemotherapy was given to 352 577 bovine-times; chemical treatment was conducted in an area of 141 660.87 hm2, including an actual mollusciciding area of 75 308.26 hm2, and environmental improvements were performed in an area of 4 738.37 hm2 in China. Data from the 453 national schistosomiasis surveillance sites of China showed that the mean Schistosoma japonicum infection rates were 0.001 5% and zero in humans and bovines in 2018, respectively, and no infected snails were found. The results demonstrate that the endemic situation of schistosomiasis appears a tendency towards a continuous decline in China; however, there is still a risk of schistosomiasis transmission, and challenges remain in achieving the target set in the Thirteenth Five-Year National Plan for Schistosomiasis Control in 2020 in some regions.

Ligation-mediated PCR for quantitative in vivo footprinting.

Ligation-mediated polymerase chain reaction (LM-PCR) is a genomic analysis technique for determination of (1) primary DNA nucleotide sequences (2) cytosine methylation patterns (3) DNA lesion formation and repair, and (4) in vivo protein-DNA footprints. However, LM-PCR can be limited by the multiple steps required and the relatively short stretch of sequence (usually <200 bp) that can be analyzed per reaction. We report here a simplified, one-day LM-PCR protocol in which all pipetting steps can be performed by a robotic workstation and which, moreover, provides longer reads (>350 bp) and enhanced signal quality by use of nonradioactive detection and a LI-COR DNA sequencing instrument. Sensitivity comparable to radiolabeling is achieved using oligonucleotide primers that are 5'-end labeled with infrared fluorochromes. We showed that the technique could be used for sensitive and reproducible in vivo photofootprinting of the human phosphoglycerate kinase 1 (PGK1) promoter, as well as providing good Maxam-Gilbert sequence information. The methods described here should allow high-throughput, high-resolution analysis of transcription factor binding and chromatin structure, and also may be useful for sequencing gaps that are refractory to cloning.

Ligation-mediated PCR: robotic liquid handling for DNA damage and repair.

The investigation of in vivo DNA repair in mammalian cells at nucleotide resolution requires the quantification of break frequencies less than one per kilobase. By optimizing several parameters of the ligation-mediated PCR technique, we find that the required sensitivity can be achieved. We also report details of a one-day procedure that can be performed either with or without a robotic liquid handling workstation. The use of near-infrared fluorescent-labeled primers with detection by a LI-COR DNA sequencer provides for safe, nonradioactive detection, similar in sensitivity to the use of 32P-labeled primers but with the additional advantage that high-quality digitized data are obtained directly. Multiplexing can be performed; that is, more than one sequence can be analyzed in a single reaction, and multiple reactions can be processed robotically. Primer sets for exons 5-8 of the tumor suppressor gene, p53, were designed for simultaneous thermal cycling. The improved procedure with infrared detection was used to monitor low-frequency damage (<1 break/kb) and/or repair of UVB, UVC, and chemical methylation. Quantitative data on the linearity of response and reproducibility are described. The coefficient of variation for technical replicates was typically 10%. The methods described here will permit high sample throughput for the detection of DNA damage and repair as well as in vivo protein footprints.

In vitro and in vivo proteolysis of the Bacillus thuringiensis subsp. israelensis CryIVD protein by Culex quinquefasciatus larval midgut proteases.

Proteases with trypsin-, chymotrypsin- and thermolysin-like specificity were detected in Culex quinquefasciatus larval midguts. Their activities were monitored by N-terminal amino acid sequence analysis of the Bacillus thuringiensis subsp. israelensis CryIVD toxin proteolytic fragments. These proteases are located in the larval midgut and in different fractions obtained during the preparation of brush border membrane vesicles. The activity of the midgut proteases increased with an increase in pH. Both the chymotrypsin- and thermolysin-like activities are involved in the processing of solubilized CryIVD toxin, whereas an additional trypsin-like protease is necessary for the CryIVD parasporal inclusion processing. The solubilized CryIVD toxin was first cleaved between Thr347 and Phe348 and between Phe348 and Tyr349, generating a 40-kDa N-terminal fragment and a 32.5-kDa C-terminal fragment. The C-terminal domain was resistant to further processing, with only a small amount of a 31-kDa product appearing due to the action of a thermolysin-like protease. However, the N-terminal domain was very unstable, and was further degraded to about 30 kDa. Unlike the solubilized CryIVD toxin, the processing of the CryIVD parasporal inclusion was very slow at neutral pH. Three protease-resistant products were detected at pHs higher than 9.5 with an overnight incubation at 37 degrees C. The 30- and 28.5-kDa C-terminal peptides are proteolytic products of trypsin- and chymotrypsin-like proteases, respectively; while the 28-kDa N-terminal peptide has 27 amino acids deleted from the N-terminal end by a thermolysin-like protease.

HMGB1-LPS complex promotes transformation of osteoarthritis synovial fibroblasts to a rheumatoid arthritis synovial fibroblast-like phenotype.

It is generally believed that some inflammatory antigens can recognize Toll-like receptors on synovial fibroblasts (SFs) and then activate downstream signals, leading to the formation of RASFs and inducing rheumatoid arthritis (RA). The objective of the current work was to study on the hypothesis that outer PAMP (LPS) binds to the inner DAMP (HMGB1) and becomes a complex that recognizes TLRs/RAGE on SFs, thus initiating a signaling cascade that leads to the secretion of inflammatory cytokines and chemokines, production of tissue-destructive enzymes, and formation of RASFs, finally resulting in RA. Osteoarthritis synovial fibroblasts (OASFs) were co-cultured with HMGB1-LPS complex in vitro for five generations to induce the transformation of human SFs to RA-like SFs (tOASFs). Then, changes of tOASFs in cell cycle and apoptosis-autophagy balance were investigated in vitro, and the pathogenicity of tOASFs was evaluated in a SCID mouse model in vivo. In vitro cell cycle analysis showed more tOASFs passing through the G1/S checkpoint and moving to S or G2 phase. Flow cytometry and confocal microscopy showed that apoptosis was reduced and autophagy was enhanced significantly in tOASFs as compared with those in OASFs. The expression of certain receptors and adhesion molecules in tOASFs was upregulated. In vivo experiments showed that tOASFs attached to, invaded, and degraded the co-implanted cartilage. In addition, histochemistry showed excessive proliferation of tOASFs and the expression of matrix metalloproteinases (MMPs). Based on the above findings, we conclude that HMGB1-LPS complex could promote the formation of RASFs.

Implication of interleukin 18 in production of matrix metalloproteinases in articular chondrocytes in arthritis: direct effect on chondrocytes may not be pivotal.

OBJECTIVE: To clarify the effect of interleukin (IL) 18 on cartilage degeneration by studying the profile of IL18 receptor (IL18R) on chondrocytes and the direct effect of IL18 on production of matrix metalloproteinases (MMPs), aggrecanases, and tissue inhibitors of metalloproteinases (TIMPs) in articular chondrocytes. METHODS: Monolayer cultured human articular chondrocytes were isolated from non-arthritic subjects and patients with rheumatoid arthritis or osteoarthritis. Gene expression of IL18, IL18Ralpha, IL18Rbeta, MMPs, and aggrecanases was detected by RT-PCR. Protein levels of IL18Ralpha were analysed by flow cytometry. Protein levels of IL18, MMPs, and TIMPs were measured by ELISA. Aggrecanase-2 mRNA expression was quantitatively analysed by real time RT-PCR. Protein levels of signalling molecules were assayed by western blotting. RESULTS: IL18 mRNA was constitutively expressed in chondrocytes, and was enhanced by IL1beta stimulation. Flow cytometric analysis showed that IL1beta, tumour necrosis factor alpha, and IL18 up regulated IL18Ralpha expression levels. The level of IL18Rbeta mRNA was much lower than that of IL18Ralpha, and was slightly up regulated by IL1beta. In chondrocytes responding to IL18, IL18 (1-100 ng/ml) slightly increased the production of MMP-1, MMP-3, and MMP-13, which was blocked by NF-kappaB inhibitor and p38 mitogen activated protein kinase inhibitor. IL18 up regulated mRNA expression of aggrecanase-2, but not aggrecanase-1. IL18 also slightly stimulated TIMP-1 production?through extracellular signal regulated kinase activation. CONCLUSION: IL18 induces production of MMPs from chondrocytes in inflammatory arthritis. Although the direct effect of IL18 on chondrocytes may not be pivotal for the induction of cartilage degeneration, IL18 seems to play some part in the degradation of articular cartilage in arthritis.

Interleukin (IL) 18 stimulates osteoclast formation through synovial T cells in rheumatoid arthritis: comparison with IL1 beta and tumour necrosis factor alpha.

OBJECTIVE: To determine whether IL18 has any indirect effects on osteoclastogenesis mediated by T cells in RA synovium, and compare its effects with those of IL1 beta and TNF alpha. METHODS: Resting T cells were isolated from peripheral blood of healthy donors, and stimulated with 2 microg/ml phytohaemagglutinin (PHA) and 0.5 ng/ml IL2 for 24 hours. Synovial T cells were isolated from RA synovial tissue. The levels of soluble receptor activator of the NF-kappa B ligand (RANKL), osteoprotegerin (OPG), IFN gamma, M-CSF, and GM-CSF were determined by ELISA. Membrane bound RANKL expression was analysed by flow cytometry. Commercially available human osteoclast precursors were cocultured with T cells to induce osteoclast formation, which was determined with tartrate resistant acid phosphatase staining and pit formation assay. RESULTS: In PHA prestimulated T cells or RA synovial T cells, IL18, IL1 beta, or TNFalpha increased soluble RANKL production and membrane bound RANKL expression in a dose dependent manner. IL18, IL1 beta, and TNF alpha did not induce M-CSF, GM-CSF, IFN gamma, or OPG production in PHA prestimulated T cells or RA synovial T cells. IL18 increased the number of osteoclasts and bone resorption area on dentine slices in the coculture of human osteoclast precursors with PHA prestimulated T cells or RA synovial T cells; its ability was equivalent to that of IL1 beta, but less potent than that of TNF alpha. In the coculture system, OPG completely blocked osteoclast induction by IL18 or IL1 beta, and greatly inhibited induction by TNF alpha. CONCLUSION: IL18, IL1 beta, or TNF alpha can indirectly stimulate osteoclast formation through up regulation of RANKL production from T cells in RA synovitis; IL18 is as effective as IL1 beta, but less potent than TNF alpha.

Relationship between baroreceptor reflex function and end-organ damage in spontaneously hypertensive rats.

The purpose of this study was to further illustrate the relationship between baroreceptor reflex sensitivity (BRS) and hypertensive end-organ damage (EOD) and to test the hypothesis that impairment of BRS aggravates EOD in hypertension. We studied baroreflex-mediated changes in heart rate [expressed as baroreceptor sensitivity to heart rate control (BRS(HR))] and blood pressure [expressed as baroreceptor sensitivity to blood pressure control (BRS(BP))] in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) that were used as controls, both at the age of 50-52 wk. Rats were also instrumented to record BP, HR, and BP variability (BPV) in the conscious, unrestrained state. In SHR compared with WKY, BP and BPV were significantly increased, whereas BRS(HR) and BRS(BP) were significantly decreased. SHR had remarkable EOD when compared with WKY (EOD score: 6.3 +/- 2.5 vs. 2.9 +/- 0.8, P < 0.01). Univariate regressive analysis demonstrated that EOD score was increased with BP and BPV and decreased with BRS. In multivariate analysis, EOD score was predicted by greater systolic BP and lower BRS and HR variability. These results indicate that BRS is negatively related to BPV and EOD score, and impaired BRS might be one of the major causes for hypertensive EOD.

[Wound repair with single or double reverse tongue-shaped skin flap].

OBJECTIVE: To seek a simple, safe and effective method for the instant repair of soft-tissue defects after excision of superficial tumor, chronic ulcer and scar constructure as well as injury. METHODS: From August 1993 to October 1997, according to the location and size of the lesion, adjacent single or double reverse tongue-shaped skin flaps were designed to repair the defects of head, face, limb and shoulder in 31 cases, and the donor sites were closed directly. RESULTS: The wounds were healed by first intention and the skin flaps were all survived postoperatively. CONCLUSION: It suggests that the tongue-shaped flap have the following characteristics: abundant blood supply, high survival rate, wide application, less trauma, and adjacent tissue can be fully used. It is a simple, safe and effective method for the instant repair of soft-tissue defects.