A synergistic hydrothermal-deep eutectic solvents (DES) pretreatment for acquiring xylooligosaccharides and lignin nanoparticles from Eucommia ulmoides wood.

To achieve an effective deconstruction for preparation of xylooligosaccharides (XOS) and lignin nanoparticles (LNPs) from Eucommia ulmoides, a synergistic pretreatment was successfully developed. Herein, the hemicelluloses were preferentially dissociated in acetic acid-catalyzed hydrothermal pretreatment (HTP) for preparation of XOS, and the hydrothermally-pretreated substrate was then subjected to deep eutectic solvents (DES) delignification for fabrication of LNPs. Results showed that the optimal yield (33.88% based on xylan) of XOS is obtained under the given HTP condition (170 °C, 0.5 h). NMR characterization showed that the linkages of lignin were mainly composed of ß-O-4, ß-ß, ß-5, etc. Besides, GPC analysis showed that the molecular weight of DES lignin fractions was lower (1130-1200 g/mol) than those of corresponding parent lignin fractions (8500-9620 g/mol). Further TEM characterization indicated that the optimal LNPs fraction has a narrow size distribution and the corresponding size is ranged from 60 to 110 nm. In short, the synergistic pretreatment could be used as a green and cost-effective approach for the development of bio-based chemicals and biomaterials from Eucommia ulmoides biomass.

Geographic Authentication of Eucommia ulmoides Leaves Using Multivariate Analysis and Preliminary Study on the Compositional Response to Environment.

To explore the influences of different cultivated areas on the chemical profiles of Eucommia ulmoides leaves (EUL) and rapidly authenticate its geographical origins, 187 samples from 13 provinces in China were systematically investigated using three data fusion strategies (low, mid, and high level) combined with two discrimination model algorithms (partial least squares discrimination analysis; random forest, RF). RF models constructed by high-level data fusion with different modes of different spectral data (Fourier transform near-infrared spectrum and attenuated total reflection Fourier transform mid-infrared spectrum) were most suitable for identifying EULs from different geographical origins. The accuracy rates of calibration and validation set were 92.86% and 93.44%, respectively. In addition, climate parameters were systematically investigated the cluster difference in our study. Some interesting and novel information could be found from the clustering tree diagram of hierarchical cluster analysis. The Xinjiang Autonomous Region (Region 5) located in the high latitude area was the only region in the middle temperate zone of all sample collection areas in which the samples belonged to an individual class no matter their distance in the tree diagram. The samples were from a relatively high elevation in the Shennongjia Forest District in Hubei Province (>1200 m), which is the main difference from the samples from Xiangyang City (78 m). Thus, the sample clusters from region 9 are different from the sample clusters from other regions. The results would provide a reference for further research to those samples from the special cluster.

Targeting the CD134-CD134L interaction using anti-CD134 and/or rhCD134 fusion protein as a possible strategy to prevent lupus nephritis.

Lupus nephritis (LN) is characterized by an increased upregulation of Th1. This study was undertaken to evaluate the role of CD134 in cytokine production in peripheral blood mononuclear cells (PBMCs) from subjects with LN. Percentages of IFN-gamma- (Th1), IL-4-, and IL-10- (Th2) producing cells within the PBMC CD4+ T cell population of LN subjects were found to be higher than those of healthy subjects. Stimulation of PBMC from LN subjects with anti-CD3 epsilon mAb/rIL-2 resulted in further increases in cytokine production. Stimulation in the presence of anti-CD134 mAb resulted in reduced IL-4 and IL-10 production; however, it also resulted in increased IFN-gamma production. Stimulation in the presence of the fusion protein rhCD134:Fc resulted in decreased production of all three cytokines. The possibilities that anti-CD134 therapy may control the extent of IL-4- and IL-10-mediated damage in active LN and that rhCD134:Fc therapy may prevent occurrence of LN are discussed.

Effect of anti-CD134L mAb and CTLA4Ig on ConA-induced proliferation, Th cytokine secretion, and anti-dsDNA antibody production in spleen cells from lupus-prone BXSB mice.

We sought to evaluate the effects of combined downregulation of CD134 and cytotoxic T lymphocyte-associated antigen 4 (CTLA4) on the autoimmune process of lupus. Concanavalin A (ConA)-induced proliferation, T helper cell cytokine secretion, and anti-double stranded DNA (dsDNA) antibody production were measured in cultures of splenic lymphocytes derived from lupus-prone BXSB mice. Splenocytes from six prednisone-treated and six untreated male lupus-prone BXSB mice, as well as from six syngeneically normal C57BL/6 male mice, were stimulated with ConA. BXSB splenocytes from untreated mice were exposed to anti-CD134L mAb, CTLA4 linked to the Fc portion of IgG1 (CTLA4Ig), or both. The magnitude of splenocyte proliferation and the levels of IFN-gamma, IL-6, and anti-dsDNA antibody were: (1) significantly higher in cultures of ConA-stimulated control and other cells than in unstimulated cells, (2) similar in cultures of normal and BXSB cells treated with anti-CD134 and CTLA4Ig or prednisone and (3) significantly reduced in cultures of ConA-stimulated and unstimulated cells treated with anti-CD134L and CTLA4Ig or prednisone compared with cells treated with CD134L or CTLA4Ig alone. Like corticosteroids, anti-CD134L mAb or CTLA4Ig can inhibit T- and B-cell activation by blocking the CD134-CD134L or CD28/CTLA4-B7 co-stimulatory pathway. The combined immune intervention described herein may prove useful for the treatment of autoimmune diseases such as systemic lupus erythematosus.

[Expression of peroxisome proliferators-activated receptor gamma and alpha-smooth muscle actin in unilateral ureteral obstruction in rats].

OBJECTIVE: To evaluate the expression of peroxisome proliferators-activated receptor gamma (PPAR gamma) and alpha-smooth muscle actin (alpha-SMA) in unilateral ureteral obstruction (UUO) in rats and the relationship between them and the extent of tubulointerstitial injury. METHODS: Male SD rats were randomly subjected to either left ureteral ligation (n=30) or sham operation (n=6), and they were sacrificed on 3, 7, 14, 21 or 28 days after UUO. Sections of renal tissue were stained with hematoxylin and eosin (HE), Masson, or periodic acid-silver methenamine (PASM). The extent of tubulointerstitial injury was determined by Banff classification. Immunohistochemical staining was performed to investigate the expression of PPAR gamma and alpha-SMA in the renal tissue. RESULTS: Swelling of tubular epithelia cells, infiltration of inflammatory cells and proliferation of fibroblasts were not so obvious on days 3 and 7 after UUO. A diffuse inflammatory cells infiltration, and massive fibrous hyperplasia were detectable on days 14 and 21 after UUO. Most tubules showed serious damage and replaced by proliferative fibrous tissue on day 28. The expression of PPAR gamma was almost undetectable in sham operation group. However, it was upregulated on day 3 and peaking on day 14, and then it was slightly decreased on days 21 and 28. The expression of alpha-SMA was only found in vascular smooth muscle cells in sham operation group. It was upregulated mainly in some interstitial cells on day 3 and increased with the progression of tubulointerstitial fibrosis. CONCLUSION: PPAR gamma expression increases in rat renal tissue after UUO, but does not always correlate with the extent of the alpha-SMA expression and tubulointerstitial injury. These data suggest that increase in PPAR gamma in renal tissue may play an important role in response to inflammation and fibrosis.

[The relation between Bence Jones protein gene polymorphism and function of renal tubular-epithelial cell in multiple myeloma].

OBJECTIVE: Bence Jones protein (BJP) plays an important role in multiple myeloma (MM) renal lesion, we try to study the relation between the characteristics of BJP variable gene and the function of renal tubular-epithelial cell (TEC). METHODS: MM patients whose function of TEC was abnormal at diagnosis constituted a group damage and patients whose function of TEC was normal for a long period a group normal. We also collected MM patients and divided them into a group BJPkappa and a group BJPlambda. Total RNAs were isolated from the mononuclear cells of bone marrow and reverse transcription was carried out with an oligo dT18 primer; these cDNAs were then amplified with PCR, cloned and sequenced, the comparison and analysis of the sequences were made according to current ESBC/Gen Bank sequence directories. RESULTS: 3/5 cases of BJPlambda use V3-4 gene in the group damage, the replacement R mutation ratio of CDRS in the group damage (7.57 +/- 3.40) was higher than that in the group normal (4.25 +/- 1.90) and higher than that of FWRS in the group damage (3.29 +/- 1.25); R mutation ratio of CDRS region in the group BJPlambda (6.64 +/- 2.38) was higher than that in the group BJPkappa (4.10 +/- 2.13) and higher than that of FWRS in the group BJPlambda (2.91 +/- 0.94), R mutation ratio of CDRS both in the group BJPkappa or BJPlambda was higher than that of FWRS respectively. CONCLUSIONS: The higher ratio of renal lesion in some MM patients and BJPlambda patients is correlated with the high ratio of their variable gene R mutation which result in changing organization and physicochemical activity of BJP. BJP translated by some subtype genes may incline to injure the function of TEC.

[Risk factors affecting the long-term outcome of IgA nephropathy].

OBJECTIVE: To study the risk factors predicting long-term renal survival of IgA nephropathy in Chinese. METHODS: Clinical and pathological data of 317 patients (124 males and 193 females) with IgA nephropathy confirmed by renal biopsy in our center from January 1987 to February 2003 were reviewed retrospectively and were correlated with outcomes. A semiquantitative scoring system was used to evaluate individual pathological lesion of the kidney. Patients were followed for at least 6 months and doubling of serum creatinine level was defined as endpoint of follow-up. Renal survival was calculated by Kaplan-Meier survival analysis and risk factors of progression were analyzed by using univariate and multi-variate Cox regression models. RESULTS: The average age at renal biopsy was (30.1 +/- 10.9) years and the average duration from onset of disease to the time of biopsy was (20.1 +/- 33.7) months. Thirty-two percent of the patients had 24 h-urinary protein excretion greater than 1.0 g at the time of biopsy. Thirty-two percent of the patients had hypertension and 20.8% had renal insufficiency. Thirty-five percent of the patients were of Lee's grade IV or above and 20.5% presented with small proportion of crescent formation (usually less than 20%). Patients were followed for an average duration of (43.5 +/- 32.2) months with 39 patients (12.3%) reaching the endpoint. The 1-, 3-, 5- and 10-year renal survival was 99.5%, 93.1%, 84.5% and 60.1% respectively. Univariate Cox regression analysis revealed that longer duration of the disease before biopsy, serum creatinine > 115 micromol/L, proteinuria > 1.0 g/d, hypertension, Lee's grading of IV-V, moderate-severe glomerulosclerosis, crescent formation, moderate-severe interstitial fibrosis and renal arteriolar lesion were risk factors of disease progression, with an odds ratio of 1.007, 9.61, 7.31, 3.97, 5.41, 5.78, 4.65, 14.05 and 2.28 respectively (P < 0.001). Episodic macro-hematuria had an odds ratio of 0.194 (P < 0.05). Age, sex, serum cholesterol and triglyceride level had no significant impact on prognosis. Proteinuria, elevated serum creatinine, glomerulosclerosis, crescent formation and interstitial fibrosis were confirmed to be independent risk factors by multi-variate Cox regression model while the remaining variables were not statistically significant. Patients with both renal insufficiency and proteinuria greater that 1.0 g/24 h at the time of biopsy had a very poor 5-year renal survival (41.8%). CONCLUSIONS: Proteinuria, renal insufficiency, glomerulosclerosis, crescent formation and interstitial fibrosis were independent risk factors predicting the renal survival. IgA nephropathy presented with proteinuria, hypertension and crescent formation may need intervention.

[Significance of calcineurin activation and CD40L expression in patients with active lupus nephritis].

AIM: To investigate the significance of the calcineurin (CaN) activation in active lupus nephritis patient. METHODS: Peripheral blood mononuclear cells (PBMCs) were separated from twenty-one active LN patients and 12 healthy controls. Phosphatase activity of CaN was determined using the CaN assay kit by measuring the content of released PO4. Reverse transcription-PCR was used to detect the expression of CD40L mRNA. Flow cytometry analysis was used to detect the expression of CD40L in LN PBMC. RESULTS: (1) Increased activation of CaN in spontaneous cultured PBMC in active LN group was found as compared with control group (46.08 +/- 5.58 vs 8.81 +/- 3.61, P < 0.01). In stimulated by PMA/Ionomycin , activity of CaN in active LN group was also higher than that of control (69.34 +/- 12.59 vs 37.12 +/- 11.57, P < 0.01). (2) Relative content of CD40L in PBMC in active LN groups increased significantly as compared with the control groups under spontaneous and PMA/Ionomycin-induced culture, respectively (P < 0.01). (3) FK506 reduced significantly production of CD40L in spontaneous and PMA/Ionomycin-induced PBMC of LN. CONCLUSION: Elevated activation of CaN in active LN may participate in regulation overexpression of CD40L in PBMC of LN. Through inhibiting CaN activity, FK506 may prevent abnormal activation of CD40-CD40L costimulatory pathway in lupus nephritis.

[Role of Akt/NF-kappa B signal transduction pathway in murine glomerular mesangial cell proliferation induced by immune complex].

AIM: To explore whether immune complex (IC) can directly induce glomerular mesangial cells(MCs) proliferation and the role of Akt/NF-kappa B signal pathway in the proliferation. METHODS: The mice were divided into control, stimulation and oligodeoxynucleotide(ODN) groups. In ODN group, MCs isolated from mice were transfected with Akt1 sense, mismatched or antisense ODN for 8 h, respectively, by using lipofectin, control and stimulation groups were incubated with lipofectin for 8 h. Then stimulation and ODN groups were incubated with aggregated IgG(AIgG)(a standard IC model), while the control group with monomeric IgG. MTT colorimetry was used to detect MCs proliferation. Distribution of MCs in cell cycle was analyzed by flow cytometry. Cyclin D1 mRNA and its protein expression were determined by RT-PCR and Western blot, respectively.The activity of NF-kappa B in MCs was determined by EMSA. RESULTS: AIgG activated NF-kappa B, upregulated cyclin D1 mRNA and its protein expression, and induced majority of MCs to enter S-phase in cell cycle. Akt1 antisense ODN specifically decreased AIgG-induced NF-kappa B activation, cyclin D1 mRNA and its protein expression, and then inhibited MCs to progress to S-phase and cell proliferation. Sense ODN and mismatched ODN had no such effects. CONCLUSION: IC can directly stimulate MCs proliferation through Akt/NF-kappa B signal pathway, suggesting that NF-kappa B probably be a useful molecule for targeted therapy in IC-mediated MC overproliferation.

Family-based association study showing that immunoglobulin A nephropathy is associated with the polymorphisms 2093C and 2180T in the 3' untranslated region of the Megsin gene.

Immunoglobulin A nephropathy (IgAN) is considered to be a multifactorial disease with genetic and environmental factors contributing to its pathogenesis. The genes involved in susceptibility and progression of the disease have not yet been clearly elucidated. Megsin (SERPINB7) is an important candidate gene, predominantly expressed in glomerular mesangium and upregulated in IgAN. To investigate the potential role of this and other genes in IgAN, patients with biopsy-proven IgAN were recruited, as were family members, for a family-based association study. The genotypes of the polymorphisms C2093T and C2180T within the 3' untranslated region of the gene were determined by polymerase chain reaction-restriction fragment length polymorphism and direct sequencing. The results were analyzed by transmission disequilibrium test (TDT) and haplotype relative risk (HRR). TDT analyses revealed that Megsin 2093C and 2180T alleles were significantly more transmitted from heterozygous parents to patients than expected (C2093T: 127 trios, P = 0.034, C2180T: 100 trios, P = 0.002). Extended TDT showed increased cotransmission of the 2093C and 2180T alleles (232 families, P < 0.001). HRR revealed that the 2093C and 2180T alleles were more often transmitted to patients (P = 0.014, <0.001, respectively). Genetic variation in Megsin confers susceptibility to IgAN.

[Effects of LFA-1 costimulation on proliferation and IgG production of PBMCs from lupus nephritis patients].

AIM: To study the effect of lymphocyte function associated antigen-1(LFA-1) costimulation on proliferation and immunoglobin production of peripheral blood mononuclear cells(PBMCs) form lupus nephritis(LN) patients. METHODS: 29 LN patients were enrolled in this study, and 12 healthy persons served as control. PBMCs from LN patients and healthy persons were obtained by Ficoll density gradient centrifugation, and cell proliferation was detected by (3)H-TdR incorporation. IgG content in cultural supermatant was detected by ELISA. RESULTS: Stimulation of anti-CD3 mAb alone could enhance the proliferation and IgG production of PBMCs from 29 LN patients,while the effects on PBMCs from patients in active phase were stronger than those from the patients in the inactive phase (P<0.01). But anti-CD3 mAb had no influenence on PBMCs from healthy persons.The costimulation of anti-CD3 mAb and LFA-1 could increase proliferation and IgG production of PBMCs from LN patients and healthy persons. The effects of this costimulation decreased in turn from active and inactive LN patients to normal persons (P<0.01). The costimulant effects of LFA-1 was inhibited by anti-LFA-1 mAb. CONCLUSION: The effects of enhancing PBMC proliferation and IgG production by LFA-1 may be a mechanism of LN pathogenesis.

[Effect of lupus recipe on IL-6 and IL-10 secretion of splenic cells in vitro in lupoid mice].

OBJECTIVE: To investigate the effect of Lupus Recipe (LR) on IL-6 and IL-10 secretion of splenic cells in vitro in lupoid model mice and on anti-dsDNA antibody. METHODS: Chronic graft-versus-host disease model was used in the experiment. The model mice were divided into four groups, the model group was un-treated and the other three groups treated with LR, prednisone and combined treatment (prednisone + LR) respectively. The serum level of ds-DNA antibody, the ConA induced splenic cell proliferation in mice's splenic cell culture as well as the IL-6, IL-10 level in the supernatant of culture were determined after treatment and compared with those of normal controls. RESULTS: (1) The splenic cell proliferative reaction in the model group splenic cells was obviously higher than that of the normal control (P < 0.05); but that in the three treated groups was different from the control insignificantly (P > 0.05); (2) The serum anti-dsDNA in the model group was higher than that in the normal control, 1.75 +/- 0.25 vs 1.20 +/- 0.21 (P < 0.01), while the difference in comparison of the treated groups with the normal control was insignificant, (P > 0.05); (3) Splenic cell IL-6 and IL-10 secretion in the model group induced by ConA was higher than those in the treated groups and the controls significantly (P < 0.05). CONCLUSION: LR reveals the effect of immunosuppressor, which could inhibit the activation of T- and B-cells, reduce the Th2 cytokine formation and auto-antibody production so as to treat lupus erythematosus effectively.