[Changes in Soil Nitrogen Components and Their Relationship with Environmental Factors with Different Tea Plantation Ages].

Changes in soil nitrogen components in tea gardens affect the soil nitrogen supply capacity and nitrogen cycle. In this study, soil samples were collected from forest land, cultivated land, and tea gardens with different plantation ages (30, 50, and 70 years) to explore the changes in soil nitrogen components and their relationship with physicochemical properties and enzyme activities. The results showed that:① with the increase in tea plantation age, the silt, total phosphorus, and urease and catalase activities gradually increased, whereas the sand, clay, pH, electrical conductivity, soil organic carbon, and the activities of invertase gradually decreased. The alkaline phosphatase activity increased first and then decreased with the increase in tea plantation age, and no significant differences were observed in soil water content and acid phosphatase activity. ② With the increase in tea plantation age, the contents of acid ammonia nitrogen, amino acid nitrogen, and nitrate nitrogen (NO3--N) increased significantly, and the contents of total nitrogen, acid ammonia nitrogen, hydrolyzable unknown nitrogen, and non-hydrolyzable nitrogen in tea gardens were significantly higher than those in forest land. ③ The total phosphorus, alkaline phosphatase, and urease were the main factors affecting soil nitrogen components. Among them, organic nitrogen components were significantly correlated with total phosphorus and alkaline phosphatase, and inorganic nitrogen components were significantly correlated with alkaline phosphatase, whereas total nitrogen had significant correlations with sand, silt, total phosphorus, urease, and alkaline phosphatase.

[Prediction of Spatial Distribution of Heavy Metals in Cultivated Soil Based on Multi-source Auxiliary Variables and Random Forest Model].

Spatial prediction of the concentrations of soil heavy metals (HMs) in cultivated land is critical for monitoring cultivated land contamination and ensuring sustainable eco-agriculture. In this study, 32 environmental variables from terrain, climate, soil attributes, remote-sensing information, vegetation indices, and anthropogenic activities were used as auxiliary variables, and random forest (RF), regression Kriging (RK), ordinary Kriging (OK), and multiple linear regression (MLR) models were proposed to predict the concentrations of As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn in cultivated soils. In comparison to those of RK, OK, and MLR, the RF model had the best prediction performance for As, Cd, Cr, Hg, Pb, and Zn, whereas the OK and RK models had highest prediction performance for Cu and Ni, respectively, showing that R2 was the highest, and mean absolute error (MAE) and root mean square error (RMSE) were the lowest. The prediction performance of the spatial distribution of soil HMs under different prediction methods was basically consistent. The high value areas of eight HMs concentrations were all distributed in the southern plain area. However, the RF model depicted the details of spatial prediction more prominently. Moreover, the importance ranking of influencing factors derived from the RF model indicated that the spatial variation in concentrations of the eight HMs in Lanxi City were mainly affected by the combined effects of Se, TN, pH, elevation, annual average temperature, annual average rainfall, distance from rivers, and distance from factories. Given the above, random forest models could be used as an effective method for the spatial prediction of soil heavy metals, providing scientific reference for regional soil pollution investigation, assessment, and management.

[Effects of Supplementation of Different Amendments on Soil Heavy Metals and Enzyme Activities in Coastal Saline Land].

The supplementation of soil amendments may not only improve the soil physical and chemical properties but also lead to the accumulation of heavy metals in soil. This experiment included six treatments:control (CK), organic manure (OM), polyacrylamide+organic manure (PAM+OM), straw mulching+organic manure (SM+OM), buried straw+organic manure (BS+OM), and bio-organic manure+organic manure (BM+OM) to explore the effects of different soil amendments on heavy metals and soil enzyme activities in coastal saline land and the relationship between them. The results revealed that compared with that in the CK treatment, the contents of soil Cr, Cu, Ni, and Pb exhibited an upward trend after the supplementation of soil amendments, among which the SM+OM and PAM+OM treatments had the most significant effects on the contents of soil Cr and Cu, respectively, whereas the BM+OM treatment had the most significant effects on the contents of soil Ni and Pb. Compared with those in the CK treatment, the activities of soil invertase and urease increased significantly following supplementation of soil amendments, and the BM+OM treatment had the best effect. The alkaline phosphatase activity exhibited a slightly upward trend after the supplementation of soil amendments, whereas the catalase activity did not change significantly. The redundancy analysis revealed that the first two axes cumulatively accounted for 70.3% of the variability in enzyme activities, and the importance of single soil heavy metals on soil enzyme activity was as follows:Ni>Cu>Cr>Pb.

[Effect of Spartina alterniflora Invasion on Soil C:N:P Stoichiometry in Coastal Wetland of Hangzhou Bay].

The invasion of Spartina alterniflora poses a great threat to coastal wetland ecosystems. In this study, the stoichiometric characteristics of soil carbon, nitrogen, and phosphorus under a Spartina alterniflora invasion were explored using ANOVA in a coastal wetland in Hangzhou Bay, and the driving coupling relationship between soil environmental factors and soil C:N:P stoichiometric characteristics of the coastal wetland were further explored based on the redundancy analysis (RDA), boosted regression tree (BRT), and partial least squares-structural equation (PLS-SEM) model. The results showed that:① after the invasion of Spartina alterniflora, soil N:P and total nitrogen (TN) in the wetland increased significantly, and with the increase in invasion time, TN and N:P decreased significantly, whereas soil organic carbon (SOC), C:N, and C:P increased significantly. ② The RDA model revealed that the main factors affecting the stoichiometric characteristics of topsoil C:N:P were SOC>electrical conductivity (EC)>TN in winter and SOC>bulk density (BD)>TN in summer. ③ The BRT model showed that under the invasion of Spartina alterniflora, TN was the key factor affecting soil C:N and N:P, and SOC was the key factor affecting C:P. ④ The PLS-SEM model showed that clay and water content directly affected SOC, thus affecting C:N and C:P; the clay and EC directly affected total phosphorus (TP), thus affecting N:P and C:P; and the EC directly affected TN, thus affecting C:N and N:P. In conclusion, the invasion of Spartina alterniflora had a significant impact on soil C:N:P stoichiometric characteristics in the study area. Soil physical properties and nutrient content directly or indirectly affected soil C:N:P stoichiometric characteristics to varying degrees.

USP7 reduction leads to developmental failure of mouse early embryos.

As a member of Ubiquitin-specific protease subfamily, ubiquitin specific protease 7 (USP7) has been reported to participate in a variety of cellular processes, including cell cycle, apoptosis, DNA damage response, and epigenetic modification. However, its function in preimplantation embryos is still obscure. To investigate the functions of USP7 during preimplantation embryo development, we used siRNA to degrade endogenous USP7 messenger RNA. We found that USP7 knockdown significantly decreased the development rate of mouse early embryos. Moreover, depletion of USP7 induced the accumulation of the DNA lesions and apoptotic blastomeres in early embryos. In addition, USP7 knockdown caused an abnormal H3K27me3 modification in 2-cell embryos. Overall, our results indicate that USP7 maintains genome stability perhaps via regulating H3K27me3 and DNA damage, consequently controlling the embryo quality.

[Spatial Heterogeneity of PM2.5 Concentration in Response to Land Use/Cover Conversion in the Yangtze River Delta Region].

The sustainable management direction of PM2.5 concentrations in the Yangtze River Delta region remains unclear due to regional spatial effects. This study combined the random forest model, spatial econometric model, and multi-scale geographically weighted regression model (MGWR) to explore the multi-scale spatial response of PM2.5 concentration to land use/cover conversion. The results show that:① PM2.5 concentrations in the Yangtze River Delta region from 2000 to 2018 showed four types of spatial-temporal patterns of spatially continuous aggregation, with strong regional synchronous changes. ② The relative influence of land conversion on PM2.5 concentrations showed a complex performance, and the source-sink effect of cultivated land and forest land was obvious. Neighborhood analysis indicated that the effect of surrounding aggregated land use conversion was generally more significant than that of single cells on PM2.5 concentration change, and the spatial effect was obvious. ③ PM2.5 concentration changes were mostly significantly negatively correlated with forest land and grassland conversion types and significantly positively correlated with conversion types between cropland, construction land, and water bodies. The importance ranking of the random forest model and correlation coefficient intensity indicated that the conversion between cropland-cropland (29.65%; 0.650), forest land-forest land (26.98%; 0.726), construction land-cropland (22.57%; 0.519), cropland-forestland (17.84%; 0.602), and cropland-construction land (16.34%; 0.424) contributed more to the variation in PM2.5 concentration. The spatial Durbin model revealed a significant spatial dependence of the change in PM2.5 concentration and a strong spatial spillover effect. ④ The MGWR model revealed the scale effects and non-stationary characteristics of the spatial relationships between different land use conversions acting on PM2.5 concentration change, and its spatial relationship showed strong differences in transfer types. However, the multi-models revealed that different land conversions drove the PM2.5 concentration change in different ways, so it is necessary to formulate targeted joint management strategies in a categorical and hierarchical manner.

Semen parameters in men recovered from COVID-19.

The novel coronavirus disease (COVID-19) pandemic is emerging as a global health threat and shows a higher risk for men than women. Thus far, the studies on andrological consequences of COVID-19 are limited. To ascertain the consequences of COVID-19 on sperm parameters after recovery, we recruited 41 reproductive-aged male patients who had recovered from COVID-19, and analyzed their semen parameters and serum sex hormones at a median time of 56 days after hospital discharge. For longitudinal analysis, a second sampling was obtained from 22 of the 41 patients after a median time interval of 29 days from first sampling. Compared with controls who had not suffered from COVID-19, the total sperm count, sperm concentration, and percentages of motile and progressively motile spermatozoa in the patients were significantly lower at first sampling, while sperm vitality and morphology were not affected. The total sperm count, sperm concentration, and number of motile spermatozoa per ejaculate were significantly increased and the percentage of morphologically abnormal sperm was reduced at the second sampling compared with those at first in the 22 patients examined. Though there were higher prolactin and lower progesterone levels in patients at first sampling than those in controls, no significant alterations were detected for any sex hormones examined over time following COVID-19 recovery in the 22 patients. Although it should be interpreted carefully, these findings indicate an adverse but potentially reversible consequence of COVID-19 on sperm quality.

[Impacts of Spartina alterniflora invasion on coastal wetland ecosystem: Advances and prospects].

Coastal wetland, affected by the interaction of land and marine ecosystems, is a typical fragile and sensitive zone. Spartina alterniflora is the most successful invasive species in global coastal zone, with important impact on coastal wetland ecosystems. We systematically summarized available literature, and reviewed the effects of S. alterniflora invasion on biogeochemical cycling (soil carbon, nitrogen, phosphorus cycling, and soil heavy metal migration) and biological community (microorganisms, plants, and animals) in coastal wetlands. Then, three perspectives were proposed for the future research: 1) strengthening the mechanism of the impact of S. alterniflora invasion on ecosystem health of coastal wetlands; 2) focusing on the coupling mechanism of the interaction between S. alterniflora community and coastal wetland environment in the context of global change; 3) carrying out long-term positioning monitoring to clarify the responses of coastal wetlands at different stages of S. alterniflora invasion. This review could provide guidance for the ecological utilization and management of S. alterniflora.

[Assessment of Soil Quality in Coastal Tidal Flat Reclamation Areas Based on MDS-TOPSIS Model].

Investigating the change of soil quality during reclamation can provide scientific guidance for desalinization, fertilization, etc. Soil samples were collected from natural tidal flat areas (0 years) and coastal tidal flat reclamation areas with different reclamation years (5, 30, 38, and 61 years) to assess the changes in soil quality after 60 years of reclamation. Soil quality was evaluated using a soil quality index (SQI), which was calculated by the selected minimum data set (MDS) and TOPSIS method. The ratio of clay content (CLAY), soil salt content (SSC), soil organic carbon (SOC), total potassium (TK), and bulk density (BD) were selected as soil quality indicators involving soil physical and chemical properties based on the MDS. The results showed that soil particle size tended to refine gradually with the continuous decrease of sand content and increase of silt and clay content. Simultaneously, soil water content (SWC) and soil BD decreased gradually during the reclamation period. Additionally, SSC, pH and sodium adsorption ratio (SAR) decreased significantly with increase in reclamation years, whereas the nutrients and available nutrients were accumulated gradually, such as soil organic matter (SOM), total nitrogen (TN), total phosphorus (TP), available nitrogen (AN), and available phosphorus (AP). However, C/N, TK and available potassium (AK) showed a decrease with increasing reclamation years, and the cation exchange capacity (CEC) showed a trend of first decrease and then increase. Finally, SQI values varied from 24.06 in the bare flat soils to 63.08 in the soils after 63 years of reclamation, and showed that bare flat (20.04±11.48) < reclamation for 5 years (29.33±10.65) < reclamation for 30 years (51.52±8.76) ≈ reclamation for 38 years (49.98±10.75) < reclamation for 61 years (58.37±3.15), which has gone through approximately three stages of "initial stability-rapid improvement-relative stability." Soil CLAY content had the highest obstacle degree towards SQI among the selected MDS indicators, followed by SSC, SOM, TK, and SWC, and thereby the lower clay content and higher salt content were the main obstacle factors for promotion of soil quality in reclamation areas.

Coding mutations in NUS1 contribute to Parkinson's disease.

Whole-exome sequencing has been successful in identifying genetic factors contributing to familial or sporadic Parkinson's disease (PD). However, this approach has not been applied to explore the impact of de novo mutations on PD pathogenesis. Here, we sequenced the exomes of 39 early onset patients, their parents, and 20 unaffected siblings to investigate the effects of de novo mutations on PD. We identified 12 genes with de novo mutations (MAD1L1, NUP98, PPP2CB, PKMYT1, TRIM24, CEP131, CTTNBP2, NUS1, SMPD3, MGRN1, IFI35, and RUSC2), which could be functionally relevant to PD pathogenesis. Further analyses of two independent case-control cohorts (1,852 patients and 1,565 controls in one cohort and 3,237 patients and 2,858 controls in the other) revealed that NUS1 harbors significantly more rare nonsynonymous variants (P = 1.01E-5, odds ratio = 11.3) in PD patients than in controls. Functional studies in Drosophila demonstrated that the loss of NUS1 could reduce the climbing ability, dopamine level, and number of dopaminergic neurons in 30-day-old flies and could induce apoptosis in fly brain. Together, our data suggest that de novo mutations could contribute to early onset PD pathogenesis and identify NUS1 as a candidate gene for PD.

[Response of Soil Enzyme Activities and Their Relationships with Physicochemical Properties to Different Aged Coastal Reclamation Areas, Eastern China].

Land reclamation transforms a tidal flat from a marine ecosystem to terrestrial ecosystem, which significantly changes the soil ecosystem process, affecting the soil enzyme activities. Soil enzyme is involved in almost all soil biochemical processes. Clarifying the relationship between soil physicochemical properties and soil enzyme activities can reveal the changes of soil quality after reclamation and provide a theoretical basis for the scientific regulation of the soil ecosystem. In this study, a total of 72 soil samples were collected from a coastal reclamation area with different years of reclamation (0, 7, 32, 40, and 63 a) in Rudong County. The changes in soil physicochemical properties and enzyme activities within different reclamation years were analyzed by classical statistical analysis, and the relationship between soil enzyme activities and physicochemical properties was analyzed using a redundancy analysis. The results showed that longterm land reclamation had significant effects on soil physicochemical properties and enzyme activities. pH and electrical conductivity showed a decreasing trend after reclamation, while the content of soil organic carbon, total nitrogen, and total phosphorus showed a trend of "rapidly accumulate-gradually consume-accumulate again" with increasing years of reclamation. Compared with the bare flat area (0 a), amylase, dehydrogenase, and acid phosphatase activity initially increased and then decreased with increasing years of reclamation, while urease and alkaline phosphatase activities were characterized by an increase-decrease-increase trend. Meanwhile, the enzyme activities within different reclamation ages were basically higher in surface soil than deep layers. Redundancy analysis revealed that the soil physicochemical properties explained 69.8% of the variation in soil enzyme activities, and an obvious relationship existed between pH, electronic conductivity, soil organic carbon, total nitrogen, total phosphorus, and soil enzyme activities (P<0.01). The order of importance of soil physicochemical properties to soil enzyme activities was as follows:total nitrogen > soil organic carbon > pH > total phosphorus > electronic conductivity.

Acute kidney injury associated with concomitant vancomycin and piperacillin/tazobactam administration: a systematic review and meta-analysis.

BACKGROUND: As a tricyclic glycopeptide antibiotic used to treat acute infections, Vancomycin (VAN) is often administered with piperacillin/tazobactam (PT) to treat various infections in clinical practice. However, whether the combination of these two drugs, compared to VAN alone, can cause an increased risk of acute kidney injury (AKI) remains controversial. OBJECTIVES: This study aims to identify the correlation between the development of AKI and the combined use of VAN and PT. METHODS: We conducted a meta-analysis of eight observational cohort studies (a total of 10727 participants received VAN and PT versus VAN and other ß-lactams). PubMed, Chinese Biological Medicine Database (CBM), China National Knowledge Infrastructure (CNKI) Database, Wan Fang Digital Periodicals Database (WFDP), and China Science Citation Database (CSCD) were searched through April 2017 using "vancomycin" and "piperacillin" and "tazobactam" as well as "acute kidney injury" or "acute renal failure" or "AKI" or "ARF" or "nephrotoxicity." Two reviewers extracted the data and assessed the risk of bias. RESULTS: A correlation was found between the development of AKI and concurrent use of VAN and PT compared with concomitant VAN and ß-lactams (OR 1.57; 95% CI, 1.13-2.01; I2 = 76.4%, p < 0.001). Similar findings were obtained in an analysis of studies comparing concurrent VAN and PT use with concurrent VAN and ß-lactam (cefepime) use (OR 1.50; 95% CI, 1.07-1.93; I2 = 80.5%, p < 0.001). Exclusion of fair-quality and low-quality articles did not change the results (OR 1.49; 95% CI, 1.06-1.92; I2 = 84.1%, p < 0.001). CONCLUSIONS: Regarding ß-lactam therapy in clinical practice, an elevated risk of AKI due to the combined use of VAN and PT should be considered.

Association between circulating leptin levels and multiple sclerosis: a systematic review and meta-analysis.

AIM: Leptin, synthesised by adipocytes, has been identified as a hormone that can influence inflammatory activity. Several studies have investigated leptin levels in patients with multiple sclerosis (MS), but the results are not consistent. This study aims to derive a more precise evaluation on the relationship between circulating leptin levels and MS. DESIGN: A comprehensive literature searched up to July 2017 was conducted to evaluate the association of circulating leptin levels and MS. The random-effect model was applied to calculate pooled standardised mean difference (SMD) and its 95% CI. MAIN OUTCOME MEASURES: Circulating leptin levels of patients with MS and healthy controls. RESULTS: Of 2155 studies identified, 33 met eligibility criteria and 9 studies with 645 patients with MS and 586 controls were finally included in the meta-analysis. Meta-analysis revealed that, compared with the healthy control group, the MS group had significantly higher plasma/serum leptin levels, with the SMD of 0.70% and 95% CI (0.24 to 1.15). Subgroup analyses suggested that the leptin levels of patients with MS were associated with region, age, study sample size, measurement type, gender and blood sample type. CONCLUSION: Overall, our study suggests that patients with MS have a significantly higher leptin level than in healthy controls. Further mechanism studies and longitudinal large cohort studies are still needed to further reveal the role of leptin in the pathogenesis of MS.

[The mechanism of irbesartan against diabetes induced myocardial fibrosis in rat model].

OBJECTIVE: To observe the protective effect of irbesartan on myocardial fibrosis in diabetic rats, and analyze the role of extracellular signal-regulated kinase (ERK) pathway in this protection. METHODS: Thirty two male SD rats were randomly divided into two groups:normal control group (CON, n=10), experimental group (n=22). Twenty diabetic rats which had modelled successfully were randomly divided into two groups:diabetic group (DM, n=10), irbesartan + DM group (Ir+DM, n=10). After 8 weeks, fasting blood glucose (FBG) level, body weight (BW), the ratio of heart weight/body weight (H/B) and left ventricular weight index (LVWI) were measured. The myocardial morphological and fibrotic changes were observed by Masson staining. Col I and col Ⅲ contents were evaluated using ELISA method, and the protein expressions of ERK1/2, p-ERK1/2 in heart tissue were tested by Western blot. RESULTS: Compared with CON group, in diabetic rats, the levels of FBG, H/B and LVWI were increased while BW was decreased. The contents of col I and col Ⅲ were increased as well as the protein expression of p-ERK1/2 and the ratio of p-ERK1/2/ERK1/2(P<0.05,P<0.01), which had the statistic differences, while ERK1/2 protein expression was not changed. Masson staining showed myocardial collagen was increased, arranged in disorder and uneven distribution. However, in Ir+ DM group, BW was increased obviously, H/B, LVWI, the contents of col I and col Ⅲ were decreased significantly (P<0.05,P<0.01), p-ERK1/2 protein expression and the ratio of p-ERK1/2/ERK1/2 were decreased (P<0.01), which had the statistic differences. Meanwhile myocardial morphology was improved significantly. CONCLUSIONS: Diabetes can induce the happening of myocardial fibrosis, and irbesartan can induce the damage of myocardial fibrosis through inhibitting the activation of ERK.

Whole-exome sequencing implicates UBE3D in age-related macular degeneration in East Asian populations.

Age-related macular degeneration (AMD) is a leading cause of irreversible central blindness among the elderly worldwide. We use exome sequencing to analyse nonsynonymous single-nucleotide variants (SNVs) across the whole genome of 216 neovascular AMD cases and 1,553 controls. As a follow-up validation, we evaluate 3,772 neovascular AMD cases and 6,942 controls from five independent cohorts in the East Asian population. Here we show strong evidence of an association at a novel, missense SNV, rs7739323, which is located in the ubiquitin protein ligase E3D (UBE3D) gene (Pmeta=1.46 × 10(-9), odds ratio (OR)=0.74, 95% confidence interval (CI): 0.63-0.88). Furthermore, ablation of the UBE3D protein lead to an abnormal amount of pigment granules deposited in retinal pigment epithelium microvilli area and an abnormal response on electroretinography (ERG) in UBE3D(+/-) heterozygous mice. Our findings indicate that the ubiquitin-proteasome system may play a role in the pathogenesis of neovascular AMD.

Bioequivalence evaluation of two formulations of pidotimod using a limited sampling strategy.

The aim of this study was to develop a limited sampling strategy (LSS) to assess the bioequivalence of two formulations of pidotimod. A randomized, two-way, cross-over study was conducted in healthy Chinese volunteers to compare two formulations of pidotimod. A limited sampling model was established using regression models to estimate the pharmacokinetic parameters and assess the bioequivalence of pidotimod. The model was internally validated by the Jack-knife method and graphical methods. The traditional non-compartmental method was also used to analyze the data and compared with LSS method. The results indicate that following oral administration of a single 800 mg dose, the plasma AUC(0-12 h) and C(max) of pidotimod can be predicted accurately using only two to four plasma samples. The bioequivalence assessment based on the LSS models provided results very similar to that obtained using all the observed concentration-time data points and indicate that the two pidotimod formulations were bioequivalent. A LSS method for assessing the bioequivalence of pidotimod formulations was established and proved to be applicable and accurate. This LSS method could be considered appropriate for a pidotimod bioequivalence study, providing an inexpensive cost of sampling acquisition and analysis. And the methodology presented here may also be applicable to bioequivalence evaluation of other medications.

7,3'-dimethoxy hesperetin inhibits inflammation by inducing synovial apoptosis in rats with adjuvant-induced arthritis.

Rheumatoid arthritis (RA) is an autoimmune disease characterized by pronounced inflammation and excessive synovial hyperplasia within affected joint. We previously reported 7, 3'-dimethoxy hesperetin (DMHP) as a highly anti-inflammatory active derivative of hesperidin showed apparent pro-apoptotic effect in vitro on fibroblast-like synoviocytes of rats with adjuvant arthritis (AA), an animal model of RA. Here, we investigated the therapeutic effects of DMHP on inflammation and synovial apoptosis in rats with AA in vivo. Paw swelling, arthritis index, TNF-α and IL-1ß serum levels were measured to evaluate the effect of DMHP on inflammation in AA rats. DNA ladder detection and TUNEL assay were used to investigate the pro-apoptotic effect of DMHP on synovial apoptosis in vivo. Bcl-2, Bax mRNA and protein expressions in synovium were determined by real-time Q-PCR and western blot, respectively. We found DMHP inhibited secondary hind paw swelling and arthritis index, and decreased TNF-α and IL-1ß serum levels in AA rats. Typical DNA ladder formation was found in DNA extraction of synovium from DMHP treated groups. The number of apoptotic synovial cells was elevated with DMHP treatment in TUNEL assay. DMHP markedly decreased Bcl-2 expression whereas increased Bax expression in synovium of AA rats at both transcription and protein levels. Moreover, DMHP treatment on AA rats significantly decreased the protein ratio of Bcl-2/Bax in synovium. In conclusion, DMHP has an apparent therapeutic effect on inflammation in rats with AA. Mechanisms of this effect are partly related to induction of synovial apoptosis through modulation of Bcl-2 and Bax expression.

Therapeutic effect of 7, 3'-dimethoxy hesperetin on adjuvant arthritis in rats through inhibiting JAK2-STAT3 signal pathway.

In our previous study, we have demonstrated that 7, 3'-dimethoxy hesperetin (DMHP), an active derivative of hesperidin, showed pro-apoptotic effect on synoviocytes in vitro. The present study was to investigate the potential therapeutic effect of DMHP on adjuvant arthritis (AA) in rat and its possible mechanisms. Freund's complete adjuvant was used to induce AA in rats. DMHP were administered intragastrically once a day from days 12 to 21 after AA induction. Secondary paw swelling, arthritis index, and pathological assessments were observed. IL-6 production in serum and IL-6 mRNA expression in synovium was detected by ELISA and real-time RT-PCR respectively. The expression of mRNA (JAK2, STAT3) and protein (JAK2, p-JAK2, STAT3, p-STAT3) in synovium were determined. We found that DMHP significantly inhibited hind paw swelling and arthritis index, and ameliorated pathological changes of ankle joint in AA rats. DMHP suppressed the level of IL-6 in serum and the expression of IL-6 mRNA in synovium of AA rats in a dose-dependent manner. DMHP apparently decreased mRNA expression of JAK2 and STAT3 as well as protein expression of p-JAK2 and p-STAT3 in the synovium of the AA rats. Correlation analysis indicated that p-JAK2 or p-STAT3 protein expression was highly correlated with joint damage severity. In conclusion, DMHP has a powerful therapeutic effect on AA in rats and its mechanisms might be partly related to inhibiting excessive activation of JAK2-STAT3 pathway.

7,3'-dimethoxy hesperetin induces apoptosis of fibroblast-like synoviocytes in rats with adjuvant arthritis through caspase 3 activation.

Approaches inducing fibroblast-like synoviocytes (FLS) apoptosis in rheumatoid arthritis (RA) patients have been considered as a promising strategy for treating RA. Here, adjuvant arthritis (AA) in rat was induced by complete Freund's adjuvant and FLS were separated and cultured using a tissue explant cultivation method. The apoptotic effect of 7,3'-dimethoxy hesperetin (DMHP, a highly antirheumatic active derivative of hesperidin) on AA FLS was evaluated with MTT assay, Hoechst staining and flow cytometry analysis. Bcl-2, Bax, caspase 3 gene expressions and caspase 3 activity were assayed to identify whether caspase 3 was involved in the apoptosis induced by DMHP. It was found that DMHP significantly decreased AA FLS proliferation in vitro by MTT assay. The AA FLS treated with DMHP displayed typical apoptotic characteristics including irregularity in shape, nuclear shrinkage and chromatin condensation. Flow cytometry analysis indicated that DMHP could obviously increase the AA FLS apoptosis rate. Compared with the AA-FLS control group, DMHP markedly decreased the mRNA expression of Bcl-2, whereas those of Bax and caspase 3 were increased. Moreover, DMHP significantly increased caspase 3 activity in a dose-dependent manner. In aggregate, the results demonstrate that DMHP effectively induces AA FLS apoptosis through caspase 3 activation and can be considered as a possible antirheumatic agent.