Resveratrol Attenuates Rheumatoid Arthritis Induce Neutrophil Extracellular Traps via TLR-4 Mediated Inflammation in C57BL/6 Mice.

The objective of this study was to evaluate whether RSV inhibits neutrophil extracellular traps (NETs) that induce joint hyperalgesia in C57BL/6 mice after adjuvant-induced arthritis. A subplantar injection of Freund's complete adjuvant was administered to C57BL/6 mice on day 0 for immunization in the AIA model. Resveratrol (RSV, 25 mg/kg) was administered intraperitoneally once daily starting on day 22 and continuing for two weeks. The effects of mechanical hyperalgesia and edema formation have been assessed in addition to histopathological scoring. Mice were sacrificed on day 35 to determine cytokine levels and PADI4 and COX-2 expression levels. ELISA was used to quantify neutrophil extracellular traps (NETs) along with neutrophil elastase-DNA and myeloperoxidase-DNA complexes in neutrophils. An immunohistochemical stain was performed on knee joints to determine the presence of nuclear factor kappa B p65 (NF-kappaB p65). AIA mice were found to have higher levels of NET in joints and their joint cells demonstrated an increased expression of the PADI4 gene. Treatment with RSV in AIA mice (25 mg/kg, i.p.) significantly (P<0.05) inhibited joint hyperalgesia, resulting in a significant increase in mechanical threshold, a decrease in articular edema, a decrease in the production of inflammatory cytokines, increased COX-2 expression, and a decrease in the immunostaining of NF-kappaB. Furthermore, treatment with RSV significantly reduced the amount of neutrophil elastase (NE)-DNA and MPO-DNA complexes, which were used as indicators of NET formation (P<0.05). This study indicates that RSV reduces NET production and hyperalgesia by reducing inflammation mediated by PADI4 and COX-2. According to these data, NETs contribute to joint pain and resveratrol can be used to treat pain in RA through this pathway.

Tyrosine kinase Pyk2 is involved in colonic smooth muscle contraction via the RhoA/ROCK pathway.

The contraction of gastrointestinal (GI) smooth muscles is regulated by both Ca(2+)-dependent and Ca(2+) sensitization mechanisms. Proline-rich tyrosine kinase 2 (Pyk2) is involved in the depolarization-induced contraction of vascular smooth muscle via a Ca(2+) sensitization pathway. However, the role of Pyk2 in GI smooth muscle contraction is unclear. The spontaneous contraction of colonic smooth muscle was measured by using isometric force transducers. Protein and phosphorylation levels were determined by using western blotting. Pyk2 protein was expressed in colonic tissue, and spontaneous colonic contractions were inhibited by PF-431396, a Pyk2 inhibitor, in the presence of tetrodotoxin (TTX). In cultured colonic smooth muscle cells (CSMCs), PF-431396 decreased the levels of myosin light chain (MLC20) phosphorylated at Ser19 and ROCK2 protein expression, but myosin light chain kinase (MLCK) expression was not altered. However, Y-27632, a Rho kinase inhibitor, increased phosphorylation of Pyk2 at Tyr402 and concomitantly decreased ROCK2 levels; the expression of MLCK in CSMCs did not change. The expression of P(Tyr402)-Pyk2 and ROCK2 was increased when CSMCs were treated with Ach. Pyk2 is involved in the process of colonic smooth muscle contraction through the RhoA/ROCK pathway. These pathways may provide very important targets for investigating GI motility disorders.

p57Kip2 is an unrecognized DNA damage response effector molecule that functions in tumor suppression and chemoresistance.

The DNA damage response (DDR) helps to maintain genome integrity, suppress tumorigenesis and mediate the radiotherapeutic and chemotherapeutic effects on cancer. Here we report that p57Kip2, a cyclin-dependent kinase (CDK) inhibitor implicated in the development of tumor-prone Beckwith-Wiedemann syndrome, is an effector molecule of the DNA-damage response. Genotoxic stress induces p57Kip2 expression via the bone morphogenetic protein-Smad1 and Atm-p38MAPK-Atf2 pathways in p53-proficient or -deficient cells and requires the Smad1-Atf2 complex that facilitates their recruitment to the p57Kip2 promoter. Elevated p57Kip2 induces G1/S phase cell cycle arrest but inhibits cell death in response to DNA damage and acts in parallel with p53 to suppress cell transformation and tumor formation. p57Kip2 is also upregulated in stage I and II clinical rectal tumor samples, likely due to genome instability of precancerous and/or early cancer cells. Targeting p57Kip2 in primary rectal cancer cells and tumor models resulted in increased sensitivity to doxorubicin, suggesting that p57Kip2 has a role in chemoresistance, which is consistent with its pro-survival function. These findings place p57Kip2 in DDR and uncover molecular mechanisms by which p57Kip2 suppresses tumorigenesis and causes chemoresistance.

Effects of different amounts of konjac flour inclusion in gestation diets on physio-chemical properties of diets, postprandial satiety in pregnant sows, lactation feed intake of sows and piglet performance.

This study was conducted to investigate the effects of different amounts of konjac flour (KF) inclusion in the gestation diet on the physio-chemical properties of diets, postprandial satiety in pregnant sows, lactation feed intake of sows and piglet performance during two successive reproductive cycles. Multiparous Landrace sows (n=140) were assigned randomly to one of four experimental diets, and four gestation diets were formulated to contain varying amounts of KF at 0%, 0.6%, 1.2% or 2.2%, respectively. The water binding capacity (WBC) (P<0.01), swelling capacity (P<0.01) of gestation diets, the concentration of total short chain fatty acids (P<0.05) after in vitro fermentation of gestation diets increased linearly with increasing inclusion amounts of KF. During the second reproductive cycle, increasing dietary KF linearly increased plasma concentrations of short chain fatty acids (SCFA) 4h postprandial (P<0.05) and glucagon-like peptide (GLP-1) 2h postprandial (P<0.05), but decreased the plasma concentration of cortisol (linearly, P<0.05) 1h postprandial. In addition, there was a linear decrease of the non-feeding oral behavior of gestating sows (P<0.01) when dietary KF increased. There were linear increases in lactation feed intake of sows during entire lactation period (P<0.01) with increasing amounts of KF in the gestation diet. In addition, the number of piglets weaned (linearly, P<0.01; quadratic, P=0.01), average piglet weights and litter weights on day 21 of lactation (linearly, P<0.01) increased with increasing inclusion amounts of KF. In conclusion, inclusion of dietary fiber with great WBC, swelling capacity and fermentation capacity in the gestation diet was beneficial for enhancing postprandial satiety in pregnant sows, increasing lactation feed intake and improved number of piglets weaned per litter through greater pre-weaning survival.

Establishment and characterization of a head kidney cell line from large yellow croaker Pseudosciaena crocea.

A continuous cell line, LYCK, derived from the head kidney of large yellow croaker Pseudosciaena crocea was established and characterized. The LYCK cell line multiplied well in Leibovitz's-15 (L-15) medium supplemented with 10% foetal bovine serum (FBS) at 28° C. This cell line, with a population doubling time of 29·5 h at passage 35, has been subcultured for >70 passages. Microscopically, LYCK cells were fibroblast-like. Chromosomal analysis revealed that the modal diploid chromosome number of LYCK cells was 48, which was identical to that of the P. crocea kidney. The cellular fluorescence could be observed in LYCK cells at 48 h after being transfected with pEGFP-N1 plasmid DNA, indicating that LYCK cells were competent for target gene expression in vitro. The expression of mRNA transcripts of the antiviral immune-related molecules interferon regulatory factor-3 I(ir3), interferon regulatory factor-7 (irf7), melanoma differentiation-associated protein 5 (mda5) was obviously up-regulated in LYCK cells in response to the stimulation with poly (I:C), whereas the expression of mRNA transcripts of the inflammatory cytokines tumour necrosis factor-α2 (tnfTNF-α2), interleukin-8 (il8), CC chemokine (lycCC) was up-regulated by lipopolysaccharide. These results indicated that the LYCK cell line could serve as a valuable tool for studies on immune-related gene functions in vitro.

Impaired hypoxic ventilatory response following neonatal sustained and subsequent chronic intermittent hypoxia in rats.

Neonatal chronic intermittent hypoxia (CIH) enhances the ventilatory sensitivity to acute hypoxia (acute hypoxic ventilatory response, HVR), whereas sustained hypoxia (SH) can have the opposite effect. Therefore, we investigated whether neonatal rats pre-treated with SH prior to CIH exhibit a modified HVR. Rat pups were exposed to CIH (5% O2/5min, 8h/day) between 6 and 15 days of postnatal age (P6-15) after pre-treatment with either normoxia or SH (11% O2; P1-5). Using whole-body plethysmography, the acute (5min, 10% O2) HVR at P16 (1 day post-CIH) was unchanged following CIH (67.9±6.7% above baseline) and also SH (58.8±10.5%) compared to age-matched normoxic rats (54.7±6.3%). In contrast, the HVR was attenuated (16.5±6.0%) in CIH exposed rats pre-treated with SH. These data suggest that while neonatal SH and CIH alone have little effect on the magnitude of the acute HVR, their combined effects impose a synergistic disturbance to postnatal development of the HVR. These data could provide important insight into the consequences of not maintaining adequate levels of oxygen saturation during the early neonatal period, especially in vulnerable preterm infants susceptible to frequent bouts of hypoxemic events (CIH) that are commonly associated with apnea of prematurity.

Tumor-suppressor role for the SPOP ubiquitin ligase in signal-dependent proteolysis of the oncogenic co-activator SRC-3/AIB1.

Steroid receptor co-activator-3 (SRC-3/AIB1) is an oncogene that is amplified and overexpressed in many human cancers. However, the molecular mechanisms that regulate 'activated SRC-3 oncoprotein' turnover during tumorigenesis remain to be elucidated. Here, we report that speckle-type POZ protein (SPOP), a cullin 3 (CUL3)-based ubiquitin ligase, is responsible for SRC-3 ubiquitination and proteolysis. SPOP interacts directly with an SRC-3 phospho-degron in a phosphorylation-dependent manner. Casein kinase Iɛ phosphorylates the S102 in this degron and promotes SPOP-dependent turnover of SRC-3. Short hairpin RNA knockdown and overexpression experiments substantiated that the SPOP/CUL3/Rbx1 ubiquitin ligase complex promotes SRC-3 turnover. A systematic analysis of the SPOP genomic locus revealed that a high percentage of genomic loss or loss of heterozygosity occurs at this locus in breast cancers. Furthermore, we demonstrate that restoration of SPOP expression inhibited SRC-3-mediated oncogenic signaling and tumorigenesis, thus positioning SPOP as a tumor suppressor.

Disseminated trichosporonosis in China.

A 20-year-old female patient presented with erythematous plaques on the nose which were progressively spreading to the trunk and the extremities, sometimes with erosions and scars. The patient was misdiagnosed as having seborrhoeic dermatitis and subacute cutaneous lupus erythematosus. The histopathological biopsy revealed mycotic infectious granuloma. Samples taken from skin lesions and other locations grew Trichosporon asahii in cultures. The identification was confirmed by molecular biological methods. The patient was treated successfully with liposomal amphotericin B in combination with fluconazole orally.

[Survival analysis after kidney transplantation in 1,180 cadaveric graftings].

OBJECTIVE: To analyze the outcome of twenty-two years of cadaveric kidney transplantation and the predict factors impacting on graft survival. METHODS: The data of 1,180 cadaveric kidney graftings in 1,039 patients between October of 1977 and June of 1999 were summarized by calculation of patient and graft survival rates using Kaplan-Meier product limit estimates. 13 factors that may influence on graft survival were analyzed by the log-rank and Cox model. RESULTS: The total of 1-, 5-, 10-, 15-, and 20-year patient survival rates were 93.02%, 80.25%, 67.86%, 65.34% and 65.34%, and the corresponding graft survival rates 88.6%, 68.3%, 43.8%, 43.8%, and 32.8%, respectively. Graft half-life was (13.98 +/- 0.96) years. The longest survivor was 22 years after cadaveric kidney transplantation. The quality of life after grafting in 835 alive patients was improved apparently; 95% of them had a normal graft function and 85% returned to work. The result of multivariable analysis using Cox model showed that 8 factors of patient's age, grafting time, pretransplant blood transfusion, cold ischemia time, delayed graft function, time of normal graft function, immunosuppressive regimen, and acute rejection were very important to affect the graft survival. CONCLUSION: Cadaveric kidney transplantation is a successful and effective method to rescue the patients with end-stage renal disease.

[Acoustic densitometry for quantification of renal allografts microcirculation and microstructure].

OBJECTIVE: To establish a new method for observing renal allografts microcirculation and microstructure with acoustic densitometry (integrated backscatter, contrast, second harmonics). METHODS: 27 patients with renal allografts were investigated by Doppler, IBS, contrast and second harmonics. Renal allografts failed age in 5 patients 20-61 years. Intravenous injection of SHU-508A was given, compared with pre-injection by quantitative assessment of acoustic densitometry (AD) of renal microstructure. RESULTS: There was a significant difference in the peak intensity, the area under the curve and the half-time of wash-out between pre-injection and injection contrast agent. The wish-in and wish-out curve was shown in a peak shape. The renal allografts with acute rejection were significantly lower than the control group in AD (P < 0.05). No contrast opacification of renal microstructure was present in 5 failed renal allografts and the wish-in and wish-out curve appeared in a straight line shape. CONCLUSION: AD is safe, and efficient in assessing renal allografts microstructure. It more sensitive in detecting allograft complication such as rejection than Doppler.

Abrogation of betaglycan attenuates TGF-beta-mediated inhibition of embryonic murine lung branching morphogenesis in culture.

Although betaglycan (TGF-beta type III receptor) is known to enhance TGF-beta ligand binding to its type II receptor in murine lung epithelial cell lines, the biological significance of this phenomenon in the process of lung organogenesis is not understood. Betaglycan gene expression was detected in embryonic murine lungs undergoing branching morphogenesis in ex vivo culture. Antisense betaglycan oligodeoxynucleotides (ODN) resulted in up to 56% stimulation of lung branching morphogenesis in culture, while betaglycan mRNA and protein expression levels were suppressed by 90 and 82%, respectively. Following abrogation of betaglycan expression with antisense oligodeoxynucleotide, embryonic lungs were relatively insensitive to TGF-beta: TGF-beta2 (0.5 ng/ml) and TGF-beta1 (20 ng/ml), respectively, down-regulated lung morphogenesis by 38 and 34% in control cultures, whereas TGF-beta-induced inhibition was attenuated to 13 and 26% respectively, in the presence of betaglycan antisense oligodeoxynucleotides. TGF-beta neutralizing antibodies also prevented TGF-beta-mediated inhibition of lung branching in culture, supporting the speculation that autocrine/paracrine TGF-beta signaling is minimal in the absence of betaglycan. Betaglycan was immunolocalized mainly to the epithelial cells in developing airways, a spatial distribution which overlaps with that of TGF-beta type II receptor. Furthermore, abrogation of endogenous betaglycan gene expression prevented the characteristic down-regulation of cyclin A and surfactant protein C (SP-C) mRNAs by exogenous TGF-beta ligands. These results show that betaglycan expression is essential for optimal TGF-beta signaling during embryonic lung development. We therefore conclude that the abrogation of endogenous betaglycan attenuates endogenous autocrine and/or paracrine TGF-beta-mediated negative regulation of lung organogenesis.

[The clinical effect of ATG and OKT3 on the treatment of refractory rejection after cadavaric kidney transplantation].

OBJECTIVE: To observe the effect of ATG and OKT3 on the treatment of refractory rejection, including acute and chronic rejection after cadaveric kidney transplantation. METHOD: 87 patients with refractory rejection after cadaveric kidney transplantation were treated by using ATG (13 patients) and OKT3 (74). A total dose of 1 000 mg for ATG group or 45 mg for OKT3 group was administered for each patient. RESULT: Two accelerating rejection and acute rejection were reversed and 7 in the ATG group, 90.00% (27/30) and 90.47% (38/42) in the OKT3 group respectively. A significant difference of total reverse rates was seen between the ATG and OKT3 groups (69.23% verse 90.28%, P < 0.05). In retransplant patients, the reverse rate of acute rejection by OKT3 was 91.67% (11/12). But OKT3 did not show any effect in patients with chronic rejection. CONCLUSION: OKT3 is better than ATG for reversing the refractory rejection, especially in retransplant patients, but it has no any effect on chronic rejection.

[Analysis of clinicopathologic changes of infection in dead patients after renal transplantation].

OBJECTIVE: To prevent renal transplantation patients from infection. METHOD: Autopsies of 24 patients who had died after renal transplantation were studied. RESULT: The incidence of infection was 66.7% and the lethal infective complications amounted for 50%. The most infectious agents were bacteria, fungi, tuberculosis, virus and actinomyces. CONCLUSION: Pathological features and clinical manifestations of lesions were caused by aforementioned agents. Some management can prevent renal transplantation recipients from infection.

[Clinical research of trabeculectomy on outpatients].

PURPOSE: To investigate the feasibility of Trabeculectomy on outpatients. METHODS: Postspectively compared and discussed the surgery effect and complications of trabeculectomy on 38 eyes of 27 outpatients with those on 38 eyes of 30 inpatients. All the surgeries were performed by the same surgeon in the same period. RESULT: The IOP of outpatients' group after one week, one month, three months or more are respectively 1.4800 +/- 0.5977 kPa, 1.8379 +/- 0.5517 kPa, 1.9650 +/- 0.4879 kPa, while the inpatients' group are 1.3468 +/- 0.5994 kPa, 1.7737 +/- 0.5177 kPa, 2.0144 +/- 0.5447 kPa. t-test has shown that the differences of the IOP after the surgery, the surgery effect and complications of trabeculectomy between the outpatients and the inpatients had no significance. CONCLUSION: Trabeculectomy on outpatients in hospital with satisfied facility would be effective, safe and feasible.

[Vision change and quality of life in the patients with cataract].

PURPOSE: We tried to assess the relationship between quality of life and visual impairment and restoring of patients with cataract and explore the factors which affect quality of life of patients with cataract. METHODS: The 142 cases of cataract with different visual impairment and the 33 cases of cataract prior to surgery and at 3 months after cataract extraction and intraocular lens implantation were measured by the scale of quality of life for diseases with visual impairment. RESULTS: The scores of quality of life and their subscores were significantly associated with the visual acuity of patients with cataract. The scores of quality of life between preoperation and after surgery had significantly difference. The factors which affect quality of life of patients with cataract included visual impairment, age and occupations. CONCLUSION: Visual impairment that caused by cataract would cause decline in quality of life. Cataract surgery was effective in improving visual acuity and quality of life of the patients. Measuring quality of life would afford a new method for evaluating visual impairment of cataract.

The relationship between dietary fat intake and risk of colorectal cancer: evidence from the combined analysis of 13 case-control studies.

The objective of this study was to examine the effects of the intake of dietary fat upon colorectal cancer risk in a combined analysis of data from 13 case-control studies previously conducted in populations with differing colorectal cancer rates and dietary practices. Original data records for 5,287 cases of colorectal cancer and 10,470 controls were combined. Logistic regression analysis was used to estimate odds ratios (OR) for intakes of total energy, total fat and its components, and cholesterol. Positive associations with energy intake were observed for 11 of the 13 studies. However, there was little, if any, evidence of any energy-independent effect of either total fat with ORs of 1.00, 0.95, 1.01, 1.02, and 0.92 for quintiles of residuals of total fat intake (P trend = 0.67) or for saturated fat with ORs of 1.00, 1.08, 1.06, 1.21, and 1.06 (P trend = 0.39). The analysis suggests that, among these case-control studies, there is no energy-independent association between dietary fat intake and risk of colorectal cancer. It also suggests that simple substitution of fat by other sources of calories is unlikely to reduce meaningfully the risk of colorectal cancer.

Development of the scale of quality of life for diseases with visual impairment.

PURPOSE: To describe the development of the of scale quality of life which can measure the quality of life of Chinese patients with visual impairment. METHODS: Based on a thorough literature search and consultation with ophthalmologists and public health professionals, 20 items were selected to create a scale. Fifty-seven cataract patients with vision impairment and 60 glaucoma patients with vision impairment and visual field loss were measured by the scale to evaluate the validity, reliability and responsiveness of the scale. RESULTS: The scale covered four domains of the quality of life (QOL). The criterion related validity of the scale: r = 0.6865 (P = 0.001). The test-retest reliability of the scale: r = 0.8959 (P = 0.001). Coronback's alpha was 0.9359. The variance ratio (VR) of intra-individual variance to inter-individual variance was 0.0551 for overall scores. The correlation coefficient of split-half method was 0.9553. The responsiveness: T-test, T = 5.95 (P = 0.001), effect size statistic was 1.533. CONCLUSIONS: The result demonstrates that the scale has a satisfactory validity, reliability and responsiveness. It suggests that the scale can be applied to clinical trials.