STAT3 inhibition ameliorates renal interstitial inflammation in MRL/lpr mice with diffuse proliferative lupus nephritis.

BACKGROUND AND OBJECTIVES: Acute kidney injury (AKI) is one of the most common and severe clinical syndromes of diffuse proliferative lupus nephritis (DPLN), of which poor prognosis is indicated by aggravated renal function deterioration. However, the specific therapy and mechanisms of AKI in DPLN remain to be explored. METHODS: The correlation between AKI and clinical pathological changes in DPLN patients was analyzed. Expression of STAT3 signaling was detected in MRL/lpr mice with DPLN using immunohistochemical staining and immunoblotting. Inhibition of STAT3 activation by combination therapy was assessed in MRL/lpr mice. RESULTS: Correlation analysis revealed only the interstitial leukocytes were significantly related to AKI in endocapillary DPLN patients. MRL/lpr mice treated with vehicle, which can recapitulate renal damages of DPLN patients, showed upregulation of STAT3, pSTAT3 and caspase-1 in renal cortex. FLLL32 combined with methylprednisolone therapy significantly inhibited the STAT3 activation, improved acute kidney damage, reduced the interstitial infiltration of inflammatory cells and decreased the AKI incidence in MRL/lpr mice. CONCLUSION: STAT3 activation may play an important role in the pathogenesis of DPLN and the development of AKI. Hence, STAT3 inhibition based on the combination of FLLL32 with methylprednisolone may represent a new strategy for treatment of DPLN with AKI.

Anticancer effect of crizotinib on osteosarcoma cells by targeting c-Met signaling pathway.

C-Met receptor and its ligand hepatocyte growth factor (HGF) are overexpressed in a variety of osteosarcoma cell lines and osteosarcoma pathological samples. It is suggested that c-Met/HGF plays an important role in the development of osteosarcoma. This study aimed to explore the anticancer effect of the c-Met-targeted drug crizotinib on osteosarcoma (OS) cells. The effects of crizotinib on the proliferation of osteosarcoma cells (SaOS2, MG-63 and MNNG) at different concentrations were detected by CCK8. Human osteosarcoma cell line MG-63 was used as an in vitro model to evaluate the effects of 2.5 µM crizotinib, 5.0 µM crizotinib and DMSO on cell apoptosis, cell cycle, migration and invasion. The expression of the c-Met signaling pathway in osteosarcoma cells was detected by western blot. The results showed that crizotinib inhibited the proliferation of cell lines in a concentration-dependent manner. Crizotinib significantly increased the number of apoptotic cells compared with the control group. Compared with the control group, crizotinib increased G0/G1 phase cells and decreased S phase cells. Compared with the control group, crizotinib inhibited the migration and invasion of osteosarcoma cells and decreased the expression of c-Met/Gab1/STAT5. This study will provide a promising therapeutic target and theoretical basis for the clinical application of crizotinib in osteosarcoma.

The combination of precipitative softening and ozonation as a pretreatment of ultrafiltration in flowback water treatment: performance and fouling analysis.

Ultrafiltration (UF) technology is an efficient shale gas flowback water treatment method. However, severe membrane fouling is the primary restriction on the application of UF technology. Here, we studied the impact of three pretreatments: precipitative softening (PS), precipitative softening, followed by ozonation (PS-O) and ozonation, followed by precipitative softening (O-PS), on pollutants' removal efficiencies and membrane fouling. The results showed that (1) the hardness, bacteria, scaling trend and compatibility with formation water exceeded the requirements for water reuse; (2) pretreatments effectively increased water flux and prolonged ultrafiltration membrane life, and both of them followed the order of PS-O process > O-PS process > PS process; (3) the fouling mechanism was changed from the complete blocking model to the standard blocking model by the PS process and the addition of ozonation enhanced the correlation of standard blocking model; (4) the quality of fracturing liquid prepared by the effluent treated by the PS-O-UF process was the best and satisfied the requirements of slick water. This paper indicated that the PS-O-UF process was suitable for the treatment of Changning shale gas flowback water for reuse.

Transcriptome Analysis and VIGS Identification of Key Genes Regulating Citric Acid Metabolism in Citrus.

Citrus (Citrus reticulata) is one of the world's most widely planted and highest-yielding fruit trees. Citrus fruits are rich in a variety of nutrients. The content of citric acid plays a decisive role in the flavor quality of the fruit. There is a high organic acid content in early-maturing and extra-precocious citrus varieties. Reducing the amount of organic acid after fruit ripening is significant to the citrus industry. In this study, we selected a low-acid variety, "DF4", and a high-acid variety, "WZ", as research materials. Through WGCNA analysis, two differentially expressed genes, citrate synthase (CS) and ATP citrate-pro-S-lyase (ACL), were screened out, which related to the changing citric acid. The two differentially expressed genes were preliminarily verified by constructing a virus-induced gene-silencing (VIGS) vector. The VIGS results showed that the citric acid content was negatively correlated with CS expression and positively correlated with ACL expression, while CS and ACL oppositely control citric acid and inversely regulate each other. These results provide a theoretical basis for promoting the breeding of early-maturing and low-acid citrus varieties.

IBV QX affects the antigen presentation function of BMDCs through nonstructural protein16.

The gamma-coronavirus infectious bronchitis virus (IBV) has a high mutation rate and mainly invades the respiratory mucosa, making it difficult to prevent and causing great economic losses. Nonstructural protein 16 (NSP16) of IBV QX also not only plays an indispensable role in virus invading but also might hugely influence the antigen's recognition and presentation ability of host BMDCs. Hence, our study tries to illustrate the underline mechanism of how NSP16 influences the immune function of BMDCs. Initially, we found that NSP16 of the QX strain significantly inhibited the antigen presentation ability and immune response of mouse BMDCs, which was stimulated by Poly (I:C) or AIV RNA. Besides mouse BMDCs, we also found that NSP16 of the QX strain also significantly stimulated the chicken BMDCs to activate the interferon signaling pathway. Furthermore, we preliminarily demonstrated that IBV QX NSP16 inhibits the antiviral system by affecting the antigen-presenting function of BMDCs.

Frequency of osteoporosis in Chinese patients with rheumatoid arthritis: a meta-analysis.

OBJECTIVE: To evaluate the current frequency of osteoporosis (OP) in Chinese patients with rheumatoid arthritis (RA) through meta-analysis. METHODS: The databases of PubMed, Web of Science, Cochrane Library, CNKI, Wan Fang, CBM, and VIP were searched for relevant literature regarding the occurrence of OP in Chinese patients with RA from January 1, 2000, to September 15, 2022. The literature was screened using inclusion and exclusion criteria, and qualifying articles were subjected to data extraction, quality evaluation, and meta-analysis using Stata 17.0 software. RESULTS: Altogether, 44 publications were included in this study, with a total sample size of 12,264 RA cases and 4532 patients with OP. Meta-analysis revealed that the frequency of OP in patients with RA was 37.67% [95% CI: 34.38%, 40.97%], while subgroup analysis showed that the frequency in South China was 49.43% [95% CI: 40.53%, 58.32%]. Also, the frequency in men was 29.29% [95% CI: 22.42%, 36.16%], which was significantly lower than in women (41.89% [95% CI: 36.87%, 46.90%]). The incidence rate in pre-menopausal women was only 15.19% [95% CI: 8.79%, 21.59%], much lower than the figure of 54.29% [95% CI: 45.28%, 63.30%] for post-menopausal women. Additionally, the frequency rates in the low, intermediate, and high disease activity groups were 24.52% [95% CI: 11.52%, 37.52%)], 27.67% [95% CI: 13.50%, 41.85%], and 57.96% [95% CI: 37.35%, 78.56%], respectively. CONCLUSION: The frequency of OP in the Chinese RA population is higher than the world average, and patients with RA should receive standardized anti-RA therapy at an early stage, with emphasis on bone health in postmenopausal women, patients with high disease activity or longer disease duration.

Transcriptome Analysis and HPLC Profiling of Flavonoid Biosynthesis in Citrus aurantium L. during Its Key Developmental Stages.

Citrus aurantium L. (sour orange) is a significant Chinese medicinal and fruit crop rich in flavonoids. However, the pathways and genes involved in flavonoid biosynthesis at the key developmental stages of Citrus aurantium L. are not fully understood. This study found that the total flavonoid concentration gradually decreased as the fruit developed. Additionally, it showed that neohesperidin was the main flavonoid in the early stages of sour orange fruit development. However, as the development stage progressed, naringin content increased rapidly and emerged as the main flavonoid component. From 27 cDNA libraries, RNA sequencing yielded 16.64 billion clean bases, including 8989 differentially expressed genes. We identified 74 flavonoid related unigenes mapped to the phenylalanine, tyrosine, and phenylpropanoid biosynthesis pathways. A total of 152 UDP-glucuronosyltransferase genes (UGTs) were identified from C. aurantium L. transcriptome database, in which 22 key flavonoid-correlated UGTs were divided into five main AtGT groups: E, G, I, L, M. We observed that the ethylene responsive factors (ERF) and myeloblastosis (MYB) family mainly regulated the key genes involved in flavonoid biosynthesis. Overall, our study generated extensive and detailed transcriptome data on the development of C. aurantium L. and characterized the flavonoid biosynthesis pattern during its fruit developmental stages. These results will benefit genetic modification or selection to increase the flavonoid content in sour oranges.

Prediction model of renal function recovery for primary membranous nephropathy with acute kidney injury.

BACKGROUND AND OBJECTIVES: The clinical and pathological impact factors for renal function recovery in acute kidney injury (AKI) on the progression of renal function in primary membranous nephropathy (PMN) with AKI patients have not yet been reported, we sought to investigate the factors that may influence renal function recovery and develop a nomogram model for predicting renal function recovery in PMN with AKI patients. METHODS: Two PMN with AKI cohorts from the Nephrology Department, the First Affiliated Hospital of Wenzhou Medical University during 2012-2018 and 2019-2020 were included, i.e., a derivation cohort during 2012-2018 and a validation cohort during 2019-2020. Clinical characteristics and renal pathological features were obtained. The outcome measurement was the recovery of renal function within 12 months. Lasso regression was used for clinical and pathological features selection. Prediction model was built and nomogram was plotted. Model evaluations including calibration curves were performed. RESULT: Renal function recovery was found in 72 of 124 (58.1%) patients and 41 of 72 (56.9%) patients in the derivation and validation cohorts, respectively. The prognostic nomogram model included determinants of sex, age, the comorbidity of hypertensive nephropathy, the stage of glomerular basement membrane and diuretic treatment with a reasonable concordance index of 0.773 (95%CI,0.716-0.830) in the derivation cohort and 0.773 (95%CI, 0.693-0.853) in the validation cohort. Diuretic use was a significant impact factor with decrease of renal function recovery in PMN with AKI patients. CONCLUSION: The predictive nomogram model provides useful prognostic tool for renal function recovery in PMN patients with AKI.

ACE2 Promoted by STAT3 Activation Has a Protective Role in Early-Stage Acute Kidney Injury of Murine Sepsis.

Sepsis-induced AKI (SIAKI) is the most common complication with unacceptable mortality in hospitalized and critically ill patients. The pathophysiology of the development of SIAKI is still poorly understood. Our recent work has demonstrated the role of signal transducer and activator of transcription 3 (STAT3) pathways in regulating inflammation and coagulation in sepsis. We hypothesized that STAT3 activation has a critical role in early-stage SIAKI. The early-stage SIAKI model was established in cecal ligation and puncture (CLP) mice, which recapitulates the clinical and renal pathological features of early-stage AKI patients. Brush border loss (BBL) was the specific pathological feature of acute tubular injury in early-stage AKI. The role of STAT3 signaling and angiotension system in early-stage SIAKI was evaluated. The STAT3 activation (increased pSTAT3) and increased angiotensin-converting enzyme 2 (ACE2) expressions were observed in CLP mice. The low responsive expressions of pSTAT3 and ACE2 to septic inflammation in CLP AKI mice were associated with BBL. Correlation analysis of proteins' expressions showed pSTAT3 expression was significantly positively related to ACE2 expression in CLP mice. Reduced pSTAT3 after S3I201 intervention, which blocked STAT3 phosphorylation, decreased ACE2 expression, and exacerbated tubular injury in early-stage SIAKI. Our data indicate that endogenous increase of ACE2 expression upregulated by STAT3 activation in early-stage SIAKI play protective role against acute tubular injury.

Physicochemical properties, content, composition and partial least squares models of A. trifoliata seeds oil.

Physicochemical properties, oil content, and fatty acids (FAs) composition are key for determining the value of oil crops. The aim of this study was to illustrate the potential of exploiting A. trifoliata as an edible oil crop, and establish a rapid measurement model for the A. trifoliata seeds oil (ASO) content and composition. In 130 A. trifoliata germplasms, the highest content of ASO was 51.27%, and unsaturated fatty acids (UFAs) mainly accounted for 74-78% of ASO. The partial least squares (PLS) model based on GC-MS and near-infrared spectroscopy was well-suited for the determination of ASO and UFA content; however, the PLS model for oleic acid (OA) and linoleic acid (LA) was not effective. The acid values and peroxide values for ASO also conformed to the Chinese food safety standards. Our findings will provide new insights and guidance for the use of A. trifoliata as oil crops..

Acute kidney injury in idiopathic membranous nephropathy with nephrotic syndrome.

BACKGROUND AND OBJECTIVES: The impact of acute kidney injury (AKI) on the progression of renal function in idiopathic membranous nephropathy (iMN) with nephrotic syndrome (NS) patients have not yet been reported, we sought to investigate the incidence, clinical features and prognosis of AKI in iMN with NS patients and determine clinical predictors for progression from AKI to advanced chronic kidney disease (CKD) stage. METHODS: We analyzed clinical and pathological data of iMN with NS patients retrospectively collected from Jan 2012 to Dec 2018. The primary renal endpoint was defined as persistent eGFR <45ml/min per 1.73 m2 more than 3 months. Comparisons of survival without primary renal endpoint were performed by Kaplan-Meier curves and log-rank test. Univariate and multivariate Cox proportional hazard models were constructed to determine independent variables associated with primary renal endpoint . RESULTS: 434 iMN with NS patients were enrolled. The incidence of AKI 1 stage, AKI 2 stage and AKI 3 stage was 23.1, 4.8 and 0.7% respectively. 66 (53.2%) patients with AKI had complete renal function recovery and 42 (33.9%) patients with AKI reached primary renal endpoint. Survival without primary renal endpoint was worse in AKI patients than No AKI patients (67.1 ± 5.3 and 43.7 ± 7.3% vs 99.5 ± 0.5 and 92.5 ± 4.2% at 2 and 4 years,p < 0.001). AKI was independently associated with primary renal endpoint, with an adjusted hazard ratio(HR) of 25.1 (95%CI 7.7-82.1, p < 0.001). CONCLUSIONS: AKI was usually mild and overlooked in iMN patients with NS, but it had a strong association with poor clinical outcomes and was an independent risk factor for CKD progression.

Long-term predictive value of acute kidney injury classification in diffuse proliferative lupus nephritis with acute kidney injury.

BACKGROUND: The long-term predictive ability of acute kidney injury (AKI) classification based on "Kidney Disease: Improving Global Outcomes"(KDIGO) AKI diagnosis criteria has not been clinically validated in diffuse proliferative lupus nephritis (DPLN) patients with AKI. Our objective was to assess the long-term predictive value of KDIGO AKI classification in DPLN patients with AKI. METHODS: Retrospective cohort study was conducted by reviewing medical records of biopsy-proven DPLN patients with AKI from the First Affiliated Hospital of Wenzhou Medical University between Jan 1, 2000 and Dec 31, 2014. Multivariate Cox regression and survival analysis were performed. RESULTS: One hundred sixty-seven DPLN patients were enrolled,82(49%) patients were normal renal function (No AKI), 40(24%) patients entered AKI-1 stage (AKI-1), 26(16%) patients entered AKI-2 stage (AKI-2) and 19(16%) patients entered AKI-3 stage (AKI-3). The mean follow-up of all patients was 5.1 ± 3.8 years. The patient survival without ESRD of all patients was 86% at 5 years and 79% at 10 years. The patient survival rate without ESRD at 10 yr was 94.5% for No AKI patients, 81.8% for AKI-1 patients, 44.9% for AKI-2 patients and 14.6% for AKI-3 patients. The area under the ROC curve for KDIGO AKI classification to predict the primary end point was 0.83 (95% CI: 0.73-0.93) (P < 0.001). In Cox regression analysis, AKI stage was independently associated with primary endpoint, with an adjusted hazard ratio (HR) of 3.8(95% CI 2.1-6.7, P < 0.001). CONCLUSION: Severity of AKI based on KDIGO AKI category was associated with progression to ESRD in DPLN patients. Analytical data also confirmed the good discriminative power of the KDIGO AKI classification system for predicting long-term prognosis of DPLN patients with AKI.

Salvianolic acid B (Sal B) alleviates the decreased activity induced by prednisolone acetate on osteoblasts by up-regulation of bone formation and differentiation genes.

Glucocorticoids (GCs) are widely used to treat a variety of autoimmune diseases, but long-term use can lead to osteoporosis. To elucidate the mechanism of osteoporosis caused by glucocorticoids and to find effective protective drugs/foods, osteoblasts treated by prednisolone acetate were studied and salvianolic acid B (Sal B) was added to osteoblasts. The results showed that Sal B increased the activity of ALP and stimulated the expression of ALP that had been suppressed by prednisolone acetate. To further study the mechanisms of the protective effect of Sal B on osteoblasts treated with prednisolone acetate, the effects of gene expression involved with bone formation and differentiation were studied. The results show that the mRNA and protein expression of Runx2, Osx, OCN, IGF-I, Col-I and HO-I was up-regulated by Sal B. In conclusion, by stimulating the osteoblast activity and the expression of genes related to bone formation and differentiation, Sal B had a protective effect on osteoblasts that had been treated with prednisolone acetate.

Baseline proteinuria level is associated with prognosis in idiopathic membranous nephropathy.

OBJECTIVES: This study aimed to investigate the unique prognostic, clinical, and renal histopathological characteristics of patients with idiopathic membranous nephropathy (IMN) with different levels of proteinuria. METHODS: This retrospective observational study included 190 IMN patients with low levels of proteinuria (low group), 193 IMN patients with medium levels of proteinuria (medium group), and 123 IMN patients with high levels of proteinuria (high group) treated between September 2006 and November 2015. Prognostic and baseline clinical and histopathological data were compared among the three groups. Poor prognostic events included the occurrence of a persistent 50% reduction in estimated glomerular filtration rate (eGFR), end-stage renal disease, or all-cause mortality. RESULTS: The severity of clinical symptoms and laboratory indices, such as blood pressure; extent of edema and hematuria; levels of fibrinogen, immunoglobulin (Ig)-G, complement (C)-4, total protein, albumin (ALB), and serum creatinine (SCr); and eGFR increased with increasing proteinuria (all p< .001). Based on renal histopathology, the extent of segmental sclerosis and balloon adhesion and renal interstitial lesion stage also increased in severity with increasing proteinuria (all p< .001). The Kaplan-Meier analysis showed that compared with patients with low and medium levels of proteinuria, patients with high levels of proteinuria had significantly lower cumulative poor event-free renal survival rates (p= .0039). CONCLUSIONS: Baseline proteinuria level is indicative of prognosis in IMN patients; the greater the extent of proteinuria is, the worse the prognosis.

Oridonin inhibits IL-1ß-induced inflammation in human osteoarthritis chondrocytes by activating PPAR-γ.

Osteoarthritis (OA), a progressive disease of the joints, affects millions of people worldwide. In the present study, we investigated the effects of oridonin, a diterpenoid isolated from Rabdosia rubescens, on IL-1ß-induced inflammation using human osteoarthritis chondrocytes. The results showed that oridonin significantly suppressed IL-1ß-induced MMP1, MMP3, and MMP13 production. IL-1ß-induced NO and PGE2 production, as well as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were also attenuated by oridonin. Western blot analysis demonstrated IL-1ß-induced NF-κB activation was reduced by oridonin. Furthermore, the expression of PPAR-γ was increased by oridonin in a concentration-dependent manner. PPAR-γ antagonist could reverse the anti-inflammatory activity of oridonin. The results suggested that oridonin could be a candidate agent for the treatment of OA.

Anti-Inflammatory Effects of Licochalcone A on IL-1ß-Stimulated Human Osteoarthritis Chondrocytes.

Licochalcone A (Lico A), a flavonoid found in licorice root (Glycyrrhiza glabra), has been reported to have anti-inflammatory activity. In this study, we evaluated the anti-inflammatory effects of Lico A on IL-1ß-stimulated human osteoarthritis chondrocytes and investigated the possible mechanism. Results demonstrated that Lico A treatment significantly inhibited PGE2 and NO production induced by IL-1ß. IL-1ß-induced iNOS and COX-2 expression were also inhibited by Lico A. Lico A inhibited MMP1, MMP3, and MMP13 production in IL-1ß-stimulated chondrocytes. Lico A also inhibited IL-1ß-induced phosphorylation of NF-κB p65 and IκBα. Meanwhile, Lico A was found to upregulate the expression of Nrf2 and HO-1. However, Nrf2 siRNA reversed the anti-inflammatory effects of Lico A. In conclusion, our results suggested that Lico A showed anti-inflammatory effects in IL-1ß-stimulated chondrocytes by activating Nrf2 signaling pathway.

Thymoquinone Inhibits IL-1ß-Induced Inflammation in Human Osteoarthritis Chondrocytes by Suppressing NF-κB and MAPKs Signaling Pathway.

Thymoquinone, an active ingredient isolated from Nigella sativa, has been reported to have anti-inflammatory effects. However, the anti-inflammatory effect of thymoquinone on IL-1ß-stimulated osteoarthritis chondrocytes remains unclear. In this study, we designed to investigate the anti-inflammatory effects and elucidated the underlying mechanism of thymoquinone on IL-1ß-stimulated human osteoarthritis chondrocytes. The effects of thymoquinone on inflammatory mediators COX-2, iNOS, NO, PGE2, as well as MMP-1, MMP3, MMP13 production were detected. The results demonstrated that thymoquinone concentration-dependently inhibited IL-1ß-induced COX-2, iNOS, NO, and PGE2 production. Thymoquinone also suppressed IL-1ß-induced MMP-1, MMP3, and MMP13 production. We found that thymoquinone significantly inhibited IL-1ß-induced NF-κB activation and IκBα degradation. In addition, thymoquinone was found to suppress IL-1ß-induced mitogen-activated protein kinases (MAPKs) activation. In conclusion, thymoquinone inhibited IL-1ß-induced inflammatory mediator production by inhibition of NF-κB and MAPKs signaling pathways in osteoarthritis chondrocytes. Thymoquinone may be a potential agent in the treatment of osteoarthritis.

Acute kidney injury in adult idiopathic nephrotic syndrome.

OBJECTIVE: This study aims to investigate the epidemiology, clinical and histological features, and prognosis of acute kidney injury (AKI) according to RIFLE classification in adult patients with idiopathic nephrotic syndrome. METHODS: In this retrospective study, 277 patients with idiopathic nephrotic syndrome were reviewed from June 2005 to June 2009. RESULTS: Fifty-one (18%) patients entered RIFLE class Risk (AKI-R); 24 (9%) patients entered RIFLE class Injury (AKI-I); and 20 (7%) patients entered RIFLE class Failure (AKI-F). Logistic regression analysis showed that severe hypoalbuminemia, increase in age, and being male were risk factors of AKI. Cumulative recovery rates in 3 months for groups AKI-R, AKI-I, and AKI-F were 95%, 100%, and 94%, respectively (p = 0.21). The mean time to recovery for groups AKI-R, AKI-I, and AKI-F was 20 ± 3, 25 ± 4, and 30 ± 5 days, respectively. Cumulative complete remission rates in 3 months for groups AKI-R, AKI-I, and AKI-F were 92%, 86%, and 65%, respectively (p = 0.002). The mean time to remission for groups AKI-R, AKI-I, and AKI-F was 28 ± 3, 39 ± 6, and 62 ± 8 days, respectively. CONCLUSION: AKI is not uncommon in adult idiopathic nephrotic syndrome. More severe AKI was associated with longer time of nephrotic syndrome complete remission. Renal function can recover completely in most of the patients.

[Identification of differentially expressed genes in rats and preliminary analysis in regression of vascular calcification].

OBJECTIVE: To investigate the differentially expressed genes in rat in the process of regression of vascular calcification by using the suppression subtractive hybridization (SSH). METHODS: 24 SD male rats which aged 6 weeks and specific pathogen free grade were selected and randomly divided into 3 groups (n = 8): control group, calcification group and regression group respectively. Vascular calcification model (vitamin D3 plus nicotine, VDN) were made from rats in calcification group and regression group, and rats in control group were intragastric administered with normal saline and lavaged with peanut oil. Rats were bred for 8 weeks in calcification group and control group, while rats in regression group were fed for 16 weeks. All rats were killed to measure concentration of calcium in the arterial tissue and examine the pathological lesion changes. Subtractive hybridization among vascular cDNA sequences from calcification group and regression group were established. The cDNA fragments which expressed higher or lower in regression group than those in calcification group were isolated. Differentially expressed genes with cDNA fragment were inserted into PMD18-T plasmid vector and transformed competent DH-5alpha, cDNA libraries of differentially expressed gene between calcification group and regression group were then constructed. Recombinant vectors were analyzed by colony PCR, positive genes were randomly selected for sequencing and analyzed by BLAST. 4 genes were randomly selected for RT-PCR certification combined with semi-quantitative analysis of DNA bands. RESULTS: VDN model of rats were successfully constructed. Concentration of tissue calcium in calcification group (15.34 mg/g +/- 2.51 mg/g) was significantly increased compared to that in control group (5.20 mg/g +/- 0.75 mg/g, P < 0.001), while in comparison with calcification group (15.34 mg/g +/- 2.51 mg/g), calcium in regression group was relatively lower (12.73 mg/g +/- 1.89 mg/g, P < 0.05). 28 up-regulated genes and 22 down-regulated genes were gained through sequencing and BLAST analysis among positive clones. RT-PCR validation indicated that 4 genes such as prdx3 and Ank2 had increasedly expressed in regression group than those in calcification group, the average fold change was 1.7. CONCLUSION: Rat vascular calcification tissue had characteristic of active regression. Genes in relation to pyrophosphoric acid synthesis, glutamate signal peptides, anti-oxidant and ant-apoptosis were up-regulated, at the same time many genes related to ossification and oxidation activity were down-regulated in the process of calcification regression. Increased expression of calcification suppressor genes accompanying decreased expression of calcification promoting genes might be the intrinsic mechanisms which initiated the active regression of calcified tissues.