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Background: Erectile dysfunction after prostate radiation therapy remains an ongoing challenge and critical quality of life issue. Given the higher dose of radiation per fraction using stereotactic body radiation therapy (SBRT) there is concern that post-SBRT impotency would be higher than conventional radiation therapy approaches. This study sought to evaluate potency preservation and sexual function following SBRT for prostate cancer., Methods: Between February 2008 and March 2011, 216 men with clinically localized prostate cancer were treated definitively with SBRT monotherapy at Georgetown University Hospital. Potency was defined as the ability to have an erection firm enough for intercourse with or without sexual aids while sexual activity was defined as the ability to have an erection firm enough for masturbation and foreplay. Patients who received androgen deprivation therapy (ADT) were excluded from this study. Ninety-seven hormone-naïve men were identified as being potent at the initiation of therapy and were included in this review. All patients were treated to 35-36.25 Gy in 5 fractions delivered with the CyberKnife Radiosurgical System (Accuray). Prostate specific antigen (PSA) and total testosterone levels were obtained pre-treatment, every 3 months for the first year and every 6 months for the subsequent year. Sexual function was assessed with the Sexual Health Inventory for Men (SHIM), the Expanded Prostate Index Composite (EPIC)-26 and Utilization of Sexual Medication/Device questionnaires at baseline and all follow-up visits., Results: Ninety-seven men (43 low-, 50 intermediate- and 4 high-risk) at a median age of 68 years (range, 48-82 years) received SBRT. The median pre-treatment PSA was 5.9 ng/ml and the minimum follow-up was 24 months. The median pre-treatment total serum testosterone level was 11.4 nmol/L (range, 4.4-27.9 nmol/L). The median baseline SHIM was 22 and 36% of patients utilized sexual aids prior to treatment. Although potency rates declined following treatment: 100% (baseline); 68% (6 months); 62% (12 months); 57% (18 months) and 54.4% (24 months), 78% of previously potent patients had erections sufficient for sexual activity at 24 months post-treatment. Overall sexual aid utilization increased from 36% at baseline to 49% at 24 months. Average EPIC sexual scores showed a slow decline over the first two years following treatment: 77.6 (baseline); 68.7 (6 months); 63.2 (12 months); 61.9 (18 months); 59.3 (24 months). All sexual functions including orgasm declined with time. Prior to treatment, 13.4% of men felt their sexual function was a moderate to big problem which increased to 26.7% two years post treatment. Post-treatment testosterone levels gradually decreased with a median value at two year follow-up of 10.7 nmol/L. However, the average EPIC hormonal scores did not illustrate a statistically significant difference two years post-treatment. Review of the radiation doses to the penile bulb in this study, a potential marker of post-treatment sexual function, revealed that the dose was relatively low and at these low doses the percentage of the penile bulb receiving 29.5 Gy did not correlate with the development of ED., Conclusions: Men undergoing SBRT monotherapy for prostate cancer report sexual outcomes comparable to those reported for conventional radiation modalities within the first 24 months after treatment. Longer follow-up is required to confirm the durability of these findings.

The mechanism of acute lung injury (ALI) following limb ischemia-reperfusion (LIR) is not yet clear. We speculate that the unbalanced expression of angiotensin-converting enzymes (ACE and ACE2) and angiotensins [Ang II and Ang-(1-7)] in the renin-angiotensin system (RAS) is a major cause of ALI. To prove this hypothesis, pathological changes, lung edema, and permeability of wild-type mice at different time points within 12 h of reperfusion after 2 h of hind-limb ischemia were first detected by morphological method, measurements of wet-to-dry weight ratio, and bronchoalveolar lavage fluid. Meanwhile, the changes of lung ACE/ACE2 mRNA and protein expression were surveyed by the methods of real-time reverse transcription-polymerase chain reaction, Western blotting, and immunohistochemistry. Angiotensin II/Ang-(1-7) levels in the blood serum and lung tissue were measured by enzyme-linked immunosorbent assay. Then the effects of ACE2 gene insertion and deletion on the previously mentioned parameters were investigated in the mice being exposed to hind-limb 2-h ischemia and 4-h reperfusion. The results revealed that lung injuries in the wild-type mice were gradually aggravated, and the expression of ACE in lung tissue was progressively increased, whereas that of ACE2 decreased within 12 h after LIR. Unexpectedly, both Ang II and Ang-(1-7) in the lung tissue were obviously increased after LIR, showing Ang-(1-7) higher than Ang II in the early stage of reperfusion but lower than Ang II at the late stage of reperfusion. Unlike local Ang II/Ang-(1-7) changes, circulating Ang-(1-7) became greatly descending, and Ang II was markedly ascending from the start of reperfusion, corresponding to local ACE/ACE2 unbalanced expression. ACE2 transgenosis improved the imbalance of ACE/ACE2 and Ang II/Ang-(1-7) expression and alleviated lung injuries, whereas ACE2 knockout further aggravated the imbalance of ACE/ACE2 and Ang II/Ang-(1-7) expression and made lung injuries more serious in the post-LIR mice. The results indicate that the dysregulation of local and circulating RAS with increased expression of ACE/Ang II and decreased expression of ACE2/Ang-(1-7) contribute to ALI caused by LIR in mice. Maintaining RAS homeostasis through upregulating ACE2 expression may lessen lung injury, which provides a new idea for the treatment of posttraumatic ALI.

Prostate cancer (PCa) is the most common type of cancer in men in the United States, which disproportionately affects African American descents. While metastasis is the most common cause of death among PCa patients, no specific markers have been assigned to severity and ethnic biasness of the disease. MicroRNAs represent a promising new class of biomarkers owing to their inherent stability and resilience. In the present study, we investigated potential miRNAs that can be used as biomarkers and/or therapeutic targets and can provide insight into the severity and ethnic biasness of PCa. PCR array was performed in FFPE PCa tissues (5 Caucasian American and 5 African American) and selected differentially expressed miRNAs were validated by qRT-PCR, in 40 (15 CA and 25 AA) paired PCa and adjacent normal tissues. Significantly deregulated miRNAs were also analyzed in urine samples to explore their potential as non-invasive biomarker for PCa. Out of 8 miRNAs selected for validation from PCR array data, miR-205 (p<0.0001), mir-214 (p<0.0001), miR-221(p<0.001) and miR-99b (p<0.0001) were significantly downregulated in PCa tissues. ROC curve shows that all four miRNAs successfully discriminated between PCa and adjacent normal tissues. MiR-99b showed significant down regulation (p<0.01) in AA PCa tissues as compared to CA PCa tissues and might be related to the aggressiveness associated with AA population. In urine, miR-205 (p<0.05) and miR-214 (p<0.05) were significantly downregulated in PCa patients and can discriminate PCa patients from healthy individuals with 89% sensitivity and 80% specificity. In conclusion, present study showed that miR-205 and miR-214 are downregulated in PCa and may serve as potential non-invasive molecular biomarker for PCa.

Liraglutide, a long-lasting glucagon‑like peptide‑1 analogue, has been used for the treatment of patients with type 2 diabetes mellitus since 2009. In this study, we investigated the anti-diabetic effects and mechanisms of action of liraglutide in a spontaneous diabetic animal model, using KK/Upj-Ay/J (KKAy) mice. The KKAy mice were divided into 2 groups, the liraglutide group (mice were treated with 250 µg/kg/day liraglutide) and the model group (treated with an equivalent amount of normal saline). C57BL/6J mice were used as the controls (treated with an equivalent amount of normal saline). The treatment period lasted 6 weeks. During this treatment period, fasting blood glucose (FBG) levels and the body weight of the mice were measured on a weekly basis. Our results revealed that liraglutide significantly decreased FBG levels, the area under the curve following a oral glucose tolerance test and insulin tolerance test, increased serum insulin levels, reduced homeostasis model assessment of insulin resistance and increased the insulin sensitivity index. Furthermore, liraglutide ameliorated glycometabolism dysfunction by increasing glycolysis via hexokinase and glycogenesis via pyruvate kinase activation. An ultrastructural examination of the pancreas revealed that liraglutide improved the damaged state of islet β cells and increased the number of insulin secretory granules. The real-time PCR results revealed that the gene expression of glucose transporter 4 (GLUT4) increased following treatment with liraglutide. Liraglutide also upregulated the protein expression of GLUT4 in liver tissue and skeletal muscle. Our results suggest that liraglutide ameliorates glycometabolism and insulin resistance in diabetic KKAy mice by stimulating insulin secretion, increasing glycogenesis and glycolysis and upregulating the expression of GLUT4.

Siganus fuscescens, which is a small commercially important marine fish, is wildly distributed in shallow waters throughout the tropical and subtropical Indo-Pacific and Eastern Mediterranean regions. It is part of a group known as rabbitfish. Fifteen new polymorphic microsatellite markers for S. fuscescens were identified, and 32 wild individuals were used to evaluate the degree of polymorphism of these markers. The number of alleles per locus ranged from 2 to 12, and the polymorphism information content ranged from 0.210 to 0.849. The observed and expected heterozygosities were 0.142-0.808 and 0.225-0.853, respectively. Although significant deviations from Hardy-Weinberg equilibrium were detected at 2 loci (Sf1-37-2 and Sf1-47), no significant deviations were detected at the other 13 loci. These microsatellite markers will provide a useful tool for studies on genetic diversity and differentiation of S. fuscescens.

Background: Microglial activations have been described in different subtypes of human prion diseases such as sporadic Creutzfeldt-Jakob disease (CJD), variant CJD, Kuru and Gerstmann-Sträussler-Scheinker disease (GSS). However, the situation of microglia in other genetic prion diseases such as fatal familial insomnia (FFI) and familial CJD remains less understood. The brain microglia was evaluated comparatively between the FFI, G114V and sCJD cases in the study., Methods: Specific Western blots, immunohistochemical and immunofluorescent assays were used to detect the changes of microglia and ELISA tests were used for levels of inflammatory cytokines., Results: Western blots, immunohistochemical and immunofluorescent assays illustrated almost unchanged microglia in the temporal lobes of FFI and G114V gCJD, but obviously increased in those of sCJD. The Iba1-levels maintained comparable in six different brain regions of FFI and G114V cases, including thalamus, cingulate gyrus, frontal cortex, parietal cortex, occipital cortex and temporal cortex. ELISA tests for inflammatory cytokines revealed significantly up-regulated IL-1β, IL-6 and TNF-α in the brain homogenates from sCJD, but not in those from FFI and G114V gCJD., Conclusion: Data here demonstrates silent brain microglia in FFI and G114V gCJD but obviously increased in sCJD, which reflects various pathogenesis of different human prion diseases subtypes.

Background: Stereotactic body radiation therapy (SBRT) delivers fewer high-dose fractions of radiation which may be radiobiologically favorable to conventional low-dose fractions commonly used for prostate cancer radiotherapy. We report our early experience using SBRT for localized prostate cancer., Methods: Patients treated with SBRT from June 2008 to May 2010 at Georgetown University Hospital for localized prostate carcinoma, with or without the use of androgen deprivation therapy (ADT), were included in this retrospective review of data that was prospectively collected in an institutional database. Treatment was delivered using the CyberKnife® with doses of 35 Gy or 36.25 Gy in 5 fractions. Biochemical control was assessed using the Phoenix definition. Toxicities were recorded and scored using the CTCAE v.3. Quality of life was assessed before and after treatment using the Short Form-12 Health Survey (SF-12), the American Urological Association Symptom Score (AUA) and Sexual Health Inventory for Men (SHIM) questionnaires. Late urinary symptom flare was defined as an AUA score ≥ 15 with an increase of  ≥ 5 points above baseline six months after the completion of SBRT., Results: One hundred patients (37 low-, 55 intermediate- and 8 high-risk according to the D'Amico classification) at a median age of 69 years (range, 48-90 years) received SBRT, with 11 patients receiving ADT. The median pre-treatment prostate-specific antigen (PSA) was 6.2 ng/ml (range, 1.9-31.6 ng/ml) and the median follow-up was 2.3 years (range, 1.4-3.5 years). At 2 years, median PSA decreased to 0.49 ng/ml (range, 0.1-1.9 ng/ml). Benign PSA bounce occurred in 31% of patients. There was one biochemical failure in a high-risk patient, yielding a two-year actuarial biochemical relapse free survival of 99%. The 2-year actuarial incidence rates of GI and GU toxicity ≥ grade 2 were 1% and 31%, respectively. A median baseline AUA symptom score of 8 significantly increased to 11 at 1 month (p=0.001), however returned to baseline at 3 months (p=0.60). Twenty one percent of patients experienced a late transient urinary symptom flare in the first two years following treatment. Of patients who were sexually potent prior to treatment, 79% maintained potency at 2 years post-treatment., Conclusions: SBRT for clinically localized prostate cancer was well tolerated, with an early biochemical response similar to other radiation therapy treatments. Benign PSA bounces were common. Late GI and GU toxicity rates were comparable to conventionally fractionated radiation therapy and brachytherapy. Late urinary symptom flares were observed but the majority resolved with conservative management. A high percentage of men who were potent prior to treatment remained potent two years following treatment.

Unlabelled: Reports suggest that Bacillus spp. can be used to increase plant growth and resistance to disease, but the molecular mechanisms underlying the interaction between Bacillus spp. and plant is not completely understood. In the present study, to clarify these underlying mechanisms, the interaction between Bacillus cereus and rice was investigated using two-dimensional gel electrophoresis. Through comparative analysis, a total of 31 differentially expressed proteins were obtained upon B. cereus NMSL88 treatment, including 22 proteins that were up-regulated and nine that were down-regulated. These data indicated that certain proteins involved in plant growth and development were up-regulated, such as xyloglucan endotransglycosylase. Interestingly, proteins involved in defence were also up-regulated, including peroxidases, glutathione S-transferases and kinases. Thus, proteins associated with disease resistance characteristics were induced in the plants after exposure to B. cereus NMSL88. In addition, several proteins involved in protein and lipid metabolism showed significant changes in expression., Significance and Impact of the Study: The present study is the first report to reveal the molecular mechanisms involved in rice seedlings in response to inoculation with Bacillus cereus at the level of proteome. The results demonstrated that B. cereus NMSL88 can up-regulate the expression of proteins related to plant growth and defence, and lead to enhanced plant growth and disease resistance., (© 2012 The Society for Applied Microbiology.)

A laboratory experiment was conducted to study the effects of different population density (D1 : 100 ind x L(-1), D2 : 150 ind x L(-1), D3 : 300 ind x L(-1)) and culture volume (V1: 50 mL, V2 : 100 mL, V3 : 400 mL) on the growth and reproduction of Moina irrasa at 25 degrees C. At the same culture density, the body length of the M. irrasa females at their first pregnancy, the first brood, and the total offsprings per female decreased with the increase of culture volumes, while the sex ratio (male/female) of the offsprings was in adverse. At the same culture volumes, the total offsprings per female decreased with the increase of culture density. At D1 V1, the body length of the females at their first pregnancy (0.95 +/- 0.10 mm) and the total offsprings (171.3 +/- 19.8 ind) per female were the maximum. At D3V2, the sex ratio was the maximum (0.54 +/- 0.05). Culture density, culture volume, and their interactions significantly affected the total offsprings per female and the sex ratio (P < 0.001).

Objective: Rheumatoid arthritis (RA) is an inflammatory and angiogenic disease. However, the molecular mechanisms that promote angiogenesis in RA have not been clearly identified. Our objective was to study the role of CD147 in angiogenesis and determine whether the strategy in which CD147 is suppressed might be useful in reducing angiogenesis in RA., Methods: Correlations among expression levels of CD147, vascular endothelial growth factor (VEGF), and hypoxia-inducible factor 1α (HIF-1α) were determined by immunohistochemistry staining. RA fibroblast-like synoviocytes (FLS) cells were cultured under various conditions, and the production of VEGF and HIF-1α was examined by real-time polymerase chain reaction and enzyme-linked immunosorbent assay. The SCID mouse coimplantation model of RA (SCID-HuRAg) was established, mice were treated with CD147 monoclonal antibody, infliximab, or both CD147 and infliximab, and the volume of the grafts and the average vascular density were measured and analyzed. Western blot analyses were performed to examine the potential signaling pathways., Results: The expression levels of CD147 showed significantly positive correlations with VEGF and HIF-1α levels, as well as with vascular density, in RA synovium. After small interfering RNA transfection or after addition of specific antibodies for CD147, the production of VEGF and HIF-1α were significantly reduced. The expression of VEGF and HIF-1α decreased more after CD147 inhibition than after infliximab treatment in the engrafted tissues in SCID-HuRAg mice. The phosphatidylinositol 3-kinase/Akt pathway may be involved in this process., Conclusion: CD147 induces up-regulation of VEGF and HIF-1α in RA FLS, further promotes angiogenesis, and leads to the persistence of synovitis. Inhibition of CD147 may be a promising target for novel therapeutic strategies., (Copyright © 2012 by the American College of Rheumatology.)

The blood CD4(+) CXCR5(+) T cells, known as "circulating" Tfh, have been shown to efficiently induce naïve B cells to produce immunoglobulin. They play an important role in certain autoimmune diseases. In the present study, we show for the first time that the frequency of CD4(+) CXCR5(+) T cells is increased in pSS patients and positively correlated with autoantibodies in the blood. The concentration of Th17-like subsets (CD4(+) CXCR5(+) CCR6(+)) in pSS patients was found to be significantly higher than in healthy controls. Functional assays showed that activated Th17-like subtypes in the blood display the key features of Tfh cells, including invariably coexpressed PD-1, ICOS, CD40L and IL-21. Th17 subsets were found to highly express Bcl-6 protein and Th1 and Th2 were not. Bcl-6 is believed to be a master transforming factor for Tfh cell differentiation and facilitate B cell proliferation and somatic hypermutation within the germinal center. These data indicate that Th17 subsets of CD4(+) CXCR5(+) T cells in the blood may participate in the antibody-related immune responses and that high frequency of CD4(+) CXCR5(+) CCR6(+) Tfh cells in blood may be suitable biomarkers for the evaluation of the active immune stage of pSS patients. It might provide insights into the pathogenesis and perhaps help researchers identify novel therapeutic targets for pSS., (Copyright © 2012 Elsevier Inc. All rights reserved.)

CdTe/CdS core(small)/shell(thick) quantum dots (QDs) with tunable near-infrared fluorescence were directly synthesized in aqueous phase through a facile one-step strategy. The QDs possessed bright fluorescence, ultrasmall size, excellent photostability and good biocompatibility. Their applicability for biological imaging was demonstrated with the in vivo active tumor targeting of nude mice.

Objective: To investigate the medication status of rheumatoid arthritis (RA) patients and to analyze the clinical use of sulphasalazine (SSZ) and the adverse effect., Methods: A total of 1 096 outpatients and inpatients diagnosed with RA were investigated in 21 hospitals all over China from July 2009 to December 2010, including gender, age of onset, clinical manifestations, as well as the clinical characteristics and medication status of 160 RA patients who received SSZ therapy., Results: In the group of 160 patients who received SSZ, the male-to-female ratio was 1:7, The average age at onset was (46.1±15.0) years, while the average course was (9.9±7.8) years. The average dose of sulphasalazine was (1.87±0.52) g/d for a mean duration of (26.3± 14.6) months. Only 17% (27/160) of the patients received SSZ monotherapy. Methotrexate (63.1%), leflunomide (36.2%) and hydroxychloroquine (18.1%) were most commonly used combination drugs. And 36.2% (58/160) of the patients used the two-drug combination of methotrexate plus sulphasalazine .In this group, 41.9% (67/160) once used SSZ but withdrew for adverse events and other reasons, while 17.5% (28/160) withdrew for adverse events, of which the most common were gastrointestinal (8.8%), skin (3.8%) and liver toxicity (3.1%)., Conclusion: Sulphaszlazine is not a common choice in the RA therapeutics in China, and the average dose of SSZ is lower than the standard dose of 2 to 3 g/d . The adverse events of SSZ are common; however, there are few severe adverse events or threat to life,SSZ is relatively safe in clinical practice.

Objective: To learn about the prevalence and risk factors of coronary artery disease (CAD) in rheumatoid arthritis (RA)., Methods: Data were obtained from a 12-month retrospective investigation of the patients with RA, randomly selected from Departments of Rheumatology and Immunology in 21 big hospitals in China. The data were collected about their social conditions, clinical conditions, medications associated with RA, such as disease modifying anti-rheumatic drugs (DMARDs), non steroidal anti-inflammatory drugs (NSAIDs), glucocorticoid, biologic agents. A nonparameter test and multivariate logistic regression analysis were performed., Results: In the study, 960 patients were enrolled. The prevalence of CAD was 3.5% in China, which was obviously higher than that of normal people. The prevalence of overweight and obesity, smoking, hypertension, diabetes mellitus, hypercholesterolemia and cerebrovascular disease were 35.1%, 12.3%, 17.0%, 7.7%, 0.4% and 3.0%, respectively. Compared with the control group, the CAD group had higher age [(64.7±9.3) years vs. (52.3±14.0) years,P<0.001], more rheumatoid nodules (14.7% vs. 3.1%,P=0.005), lower rate of hydroxychloroquine (HCQ) use (5.9% vs. 22.6%,P=0.021), higher prevalence rates of lung interstitial disease (17.5% vs. 7.0%,P<0.001), diabetes mellitus and hypertension (29.4% vs. 7.0%,P<0.001; 38.2% vs. 16.2%,P=0.001). There was no obvious correlation of CAD in RA with joint deformity, rheumatoid factor (RF) titer, glucocorticoid use, hypercholesterolemia and body mass index (BMI). Multivariate analysis showed higher age, diabetes mellitus and hypertension were independent predictors of CAD, and the use of HCQ was a protective factor of CAD., Conclusion: The prevalence of CAD is 3.5%. Higher age, diabetes mellitus and hypertension are independent predictors of CAD, and the use of HCQ is a protective factor of CAD.

Objective: To investigate the current status of tumor necrosis factor (TNF) inhibitors application in rheumatoid arthritis (RA) patients in China and to analyze the related factors., Methods: A retrospective survey was conducted in 21 hospitals from different parts of China. The patients with RA were randomly enrolled. Data of their social backgrounds, clinical conditions, usage and adverse effects of TNF inhibitors were collected. The costs of TNF inhibitors and the indirect costs of the disease were calculated. A multivariate Logistic regression analysis was performed to analyze the factors related to TNF inhibitors application., Results: In the study, 1 095 RA patients from July 2009 to November 2010 were enrolled, of whom 112 had received TNF inhibitors, representing 10.2% of the total patients. The patients who received etanercept and infliximab were 7.4% (86/1 095) of the patients and 2.4% (26/1 095), respectively. There were 0.5% of the patients (5/1 095) who had received both of the TNF inhibitors. The patients who had accepted etanercept and treatment duration for less than 3 months and 3-6 months accounted for 38.5% and 25.0% respectively, while those treated with Infliximab were 38.1%. Their health assessment questionnaire (HAQ) scores were 1.1, 0.5 and 0.1, corresponding to treatment duration of infliximab for less than 3, 3-6 and 6-9 months and those were 1.3, 1.0, 0.3 corresponding to treatment duration of etanercept, respectively. Infliximab costs were RMB 24 525.0, 69 300.0 and 96 800.0 Yuan and etanercept costs were RMB 7 394.8, 9 158.6, 54 910.9 Yuan, respectively. Indirect costs for RA patients who accepted infliximab for less than 3, 3-6 and 6-9 months were RMB 365.6, 0 and 158.9 Yuan and those who accepted etanercept were RMB 2 158.4, 288.5 and 180.1 Yuan, respectively. Allergy and infection were the main side-effects of etanercept and both happened in 3.5% of all the patients. Liver damage happened in 2.3% of all the patients, while allergy and infection happened in 6.5% of all the patients who accepted infliximab. Logistic regression analysis showed that patients with higher education experience increased the odds of entering the TNF inhibitors group (OR: 1.292, 95%CI: 1.132-1.473, P=0.000)., Conclusion: About one-tenth of RA patients in China have accepted TNF inhibitors. Higher education experience is the key factor for using TNF inhibitors.

Aim: To investigate whether monocytes activated with lipopolysaccharide(LPS) have an effect on Th17 cell differentiation in humans, CD4(+) T cell and CD14(+) monocytes activated with LPS were treated in the absence or presence of anti-CD3 mAb with various concentrations at different time points., Methods: Purification of CD4(+) T cell and CD14(+) monocytes were performed by magnetic cell sorting and cultured together. Cultures were stimulated with LPS alone or anti-CD3 mAb alone or LPS plus anti-CD3 mAb for 3 days. In the anti-CD3 mAb stimulation cells were added different concentrations of LPS. Cells were activated under LPS/anti-CD3 costimulation for 3, 6, or 10 days. The percentage of IL-17(+) T cells and INF-γ(+) T was determined by flow cytometry., Results: LPS or anti-CD3 mAb alone induced only very low levels of IL-17(+) T cells, (1.30 ± 0.19)%, (1.10 ± 0.21)%, respectively. The percentage was substantially higher in the LPS and anti-CD3 mAb costimulationa as much as(2.01 ± 0.46)%. In the presence of 0.1 μg/mL, 1 μg/mL, 10 μg/mL LPS, the proportion of Th17 reached to (1.92 ± 0.21)%, (1.30 ± 0.37)%, (1.01 ± 0.25)%. Low-concentration LPS (0.1 μg/mL) stimulation favored Th17 differentiation. The highest proportion of IL-17(+) T cells was found at day 3(2.13 ± 0.32)%, with levels declining at day 6 and day 10, while, Th1 at day 6(17.45 ± 3.04)%, declining at day 10., Conclusion: Low-concentration LPS stimulation plus anti-CD3 mAb in short term support optimal Th17 generation. Nevertheless, this model closely mimics the environment of rheumatoid arthritis in vivo and proposes an effective model for the generation of human Th17 cells.

Virus infection of plants may induce a variety of disease symptoms. However, little is known about the molecular mechanism of systemic symptom development in infected plants. Here we performed the first next-generation sequencing study to identify gene expression changes associated with disease development in tobacco plants (Nicotiana tabacum cv. Xanthi nc) induced by infection with the M strain of Cucumber mosaic virus (M-CMV). Analysis of the tobacco transcriptome by RNA-Seq identified 95,916 unigenes, 34,408 of which were new transcripts by database searches. Deep sequencing was subsequently used to compare the digital gene expression (DGE) profiles of the healthy plants with the infected plants at six sequential disease development stages, including vein clearing, mosaic, severe chlorosis, partial and complete recovery, and secondary mosaic. Thousands of differentially expressed genes were identified, and KEGG pathway analysis of these genes suggested that many biological processes, such as photosynthesis, pigment metabolism and plant-pathogen interaction, were involved in systemic symptom development. Our systematic analysis provides comprehensive transcriptomic information regarding systemic symptom development in virus-infected plants. This information will help further our understanding of the detailed mechanisms of plant responses to viral infection.

Aim: To investigate the possible relationship between the influencing factors occurring before and during birth in full-term infants and the outcome of retinopathy., Methods: Totally 816 full-term infants admitted in the neonate intensive unit of Boai Hospital of Zhongshan between 1 May, 2008 and 30 June, 2011 were included in the study. Fundus examination was performed and evaluated individually on them at the age of 48 hours after delivery, 2 weeks and 1 month. Some possible risk factors happening prenatally or during delivery such as pregnant related hypertension, placenta previa, placental abruption etc, as well as some neonatal risk factors such as neonatal asphyxia, hypoxic-ischemic encephalopathy (HIE), low birth weight etc, were recorded and evacuated. Then the effect of the risk factors of full-term infants on retinopathy was studied., Results: The incidence of retinal hemorrhage of full-term infants with prenatal pregnant related hypertension (PRH) of the mother (43.6%) was significantly higher than that of full-term infants without (8.0%). (P<0.001). The incidence of retinal hemorrhage of full-term infants with neonatal asphyxia and /or hypoxic-ischemic encephalopathy (HIE)(29.3%) was significantly higher than that of those without (15.7%), but correlation was not found between the severity of retina hemorrhage and the degree of hypoxic disease. A pale color of optic disc was associated with a low birth weight of full-term infant. Full-term infants with birth weigh less than 2500g had a significant higher incidence of retinopathy than those with birth weight equal or more than 2500g ( P<0.001)., Conclusion: The main influencing factors which lead to retinopathy of high risk full-term infants are prenatal factors such as PRH, and some neonatal risk factors such as asphyxia, hypoxic-ischemic encephalopathy, and low birth weight.

Objective: To evaluate the value of endometrial microvessel density (MVD) in assessing the endometrial receptivity during the peri-implantation period., Methods: A total of 104 patients undergoing in vitro fertilization and embryo transfer (IVF-ET) treatment were analyzed retrospectively. The subjects were divided into clinical pregnancy group (50 cases) and nonpregnant group (54 cases) according to the IVF-ET outcome. Endometrial tissues were collected 7 days after the natural ovulation prior to IVF-ET for measurement of the endometrial MVD using electron microscopy, which was analyzed in relation to the clinical outcome of the treatment., Results: The endometrial MVD was significantly higher in the clinical pregnancy group than in the nonpregnant group [(4.12∓1.84)% vs (3.46∓1.26)%, t=-2.127, P=0.036). ROC curve analysis showed that the MVD had an area under the curve slightly over 0.5 (0.598) for predicting clinical pregnancy, suggesting a poor specificity in predicting the clinical outcome of the treatment., Conclusion: In IVF-ET cycles, the endometrial MVD during the peri-implantation period is helpful for assessing the endometrial receptivity, but the specificity remains low.

Objective: To explore the efficacy of frozen-thawed embryo transfer combined with intrauterine administration of autologous peripheral blood mononuclear cells (PBMCs) in the treatment of repeated implantation failure (RIF)., Methods: PBMCs obtained from 3 patients with RIF on the day of follicle rupture (natural cycle) or when the endometrial thickness reached 8 mm (hormone replacement cycle) were cultured in the presence of HCG for 48 h. The cultured PBMCs, along with freshly isolated PBMCs, were administered into the uterine cavity of the patients. Vitrified cleavage-stage embryos or blastocysts transfer was performed on day 3 or 5, respectively., Results: Vitrified embryo or blastocyst transfer resulted in pregnancy and healthy live births in all the 3 patients., Conclusion: Frozen-thawed embryo transfer combined with intrauterine administration of autologous PBMCs may be an effective and safe approach to the treatment of RIF and may improve the outcomes of assisted reproduction.

Objective: To investigate the relationship between cell apoptosis and the quality of early mouse embryos, understand the significance of apoptosis-regulatory genes in early embryonic development, and explore a new approach to improving the embryo quality., Methods: The levels of cell apoptosis and proliferation in early mouse embryos in different developmental status (morphologically normal embryos, arrested embryos and fragmented embryos) were analyzed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), caspase in situ fluorescence and Bcl-2 immunofluorescence, and immunofluorescent detection of proliferating cell nuclear antigen (PCNA)., Results: The cells in arrested embryos and embryonic fragments showed positive results in TUNEL assay with enhanced caspase activity and lowered expressions of Bcl-2 and PCNA., Conclusion: Cell apoptosis in early mouse embryos may be closely related to embryonic arrest and fragmentation.

Objective: To investigate the value of serum estradiol increment and serum estradiol/follicles on the day of hCG administration in predicting the clinical outcomes of in vitro fertilization-embryo transfer (IVF-ET)., Methods: A retrospective analysis of the IVF-ET data was conducted involving 121 patients who received a long gonadotrophin-releasing hormone agonist (GnRHa) protocol. According to the increment of serum estradiol on the day of hCG administration (relative to the level on the day before hCG administration), the patients were divided into 3 groups (A1, A2 and A3) with a increment ratio below 30%, between 30% and 50%, and over 50%, respectively. In addition, according to the ratio of serum estradiol level on hCG day to mature follicle (diameter ≥ 14 mm) number, these patients were divided into three groups (B1, B2 and B3) with the ratio below 250 pg/ml, between 250 and 350 gp/ml, and over 350 pg/ml, respectively. The hormonal characteristics and clinical outcomes of the IVF-ET cycles were analyzed comparatively., Results: Both the clinical pregnancy rate (71.05%) and embryo implantation rate (52.63%) were significantly higher in group A3 than in groups A1 and A2 (P<0.05). The best clinical pregnancy rate (67.86%) and embryo implantation rate (49.14%) were significantly higher in group B2 than in groups B1 and B3 (P<0.05)., Conclusion: The variation of serum estradiol shows an important impact on the clinical outcomes of IVF-ET in patients receiving long GnRH-a protocol. Favorable outcomes can be expected with a hCG day serum estradiol increment ratio above 50% and E(2)/follicle ratio between 250 and 350 pg/ml.

Background: Dislocation of posterior chamber intraocular lens is one of the most common complications of intraocular lens implantation. Lens exchange is an effective solution to this unsatisfactory status. This study was conducted to analyze the possible predisposing factors for out-of-the-bag posterior chamber intraocular lens dislocation and to study the outcomes of lens exchange surgery., Methods: Thirty-six consecutive patients (36 eyes) with out-of-the-bag intraocular lens dislocation who underwent posterior chamber intraocular lens exchange in Zhongshan Ophthalmic Center of Sun Yat-sen University (Guangdong, China) from January 2003 to October 2009 were included. A 6-month follow-up was completed. The causes for out-of-the-bag intraocular lens dislocation and visual outcomes of posterior chamber intraocular lens exchange were analyzed. The out-of-the-bag intraocular lens dislocation was diagnosed on the basis of the findings from slit-lamp microscope and B-ultrasound. The dislocated intraocular lens was explanted. Reimplantation of a new posterior chamber intraocular lens was performed in each case using standardized surgical procedures., Results: In this study, a total of thirty-six consecutive patients (36 eyes) with out-of-the-bag intraocular lens dislocation underwent posterior chamber intraocular lens exchange surgery. Causes for out-of-the-bag intraocular lens dislocation included posterior capsule rupture during the initial cataract extraction procedure (23 eyes, 63.8%), trauma (5 eyes, 13.9%),neodymium-doped yttrium aluminium garnet (Nd:YAG) laser-induced dislocation (2 eyes, 5.6%), the status after vitrectomy (2 eyes, 5.6%) and unidentifiable etiology (4 eyes, 11.1%). Symptoms of these patients mainly included decrease in visual acuity (17 cases, 47.2%), blurred vision (16 cases, 44.4%), glare (1 case, 2.8%), diplopia (1 case, 2.8%), and halo (1 case, 2.8%). Intraocular lens dislocation into the posterior vitreous cavity (29 eyes, 80.5%), anterior chamber (1 eye, 2.8%) and anterior vitreous cavity (6 eyes, 16.7%) was found in this series. The foldable intraocular lenses (acrylic or silicone) were explanted from 27 eyes (75%) and rigid intraocular lenses (poly methyl methacrylate, PMMA) from 9 eyes (25%). The most common explanted intraocular lens material was single-piece acrylic (13 pieces, 36.1%), followed by 3-piece acrylic (9 pieces, 25%), single-piece PMMA (9 pieces, 25%), and 3-piece silicone (5 pieces, 13.9%). Uncorrected visual acuity postoperatively improved in 29 eyes (81%), unchanged in 4 eyes (11%), and worsened in 3 eyes (8%) in comparison to that before exchange operation (P = 0.006). Best corrected visual acuity tended to improve, but the improvement was not significant (P = 0.206). Complications related to lens exchange surgery were mainly intraocular lens redislocation (1 eye), retinal detachment (1 eye), vitreous hemorrhage (1 eye), and cystoid macular edema (1 eye)., Conclusions: Out-of-the-bag intraocular lens dislocation was mainly caused by posterior capsule rupture during the initial cataract extraction procedure and the foldable lens was the most common dislocated intraocular lens. In most cases, posterior chamber intraocular lens exchange surgery could provide satisfied final visual outcomes.

Objective: To study the clinical factors affecting the outcomes of repeated assisted reproductive technology (ART) cycles., Methods: A retrospective analysis of the clinical data and outcomes was conducted among 160 patients undergoing repeated IVF/ICSI-ET treatment between January 2006 and April 2009., Results: The patients with successful clinical pregnancy after two ART cycles (group A) had a younger age and shorter duration of infertility, and had more antral follicles (AFC), more eggs and good-quality embryos with more transferred embryos available and higher good-quality embryo rate (P<0.05) than those who failed to have pregnancy after the cycles (group B). In the second cycle, the patients in group A had higher doses of short-acting GnRHa, r-HCG and HMG and at the same time more good eggs and embryos than in the first cycle., Conclusions: Female age is one of the most important factors affecting the pregnancy rate after repeated ART cycles. The clinical pregnancy rate can be enhanced by administering short-acting GnRHa, HMG, oral contraceptives and adjusting the dose of Gn as well as changing the culture medium of embryos.

MicroRNAs (miRNAs) play important roles in regulating a plethora of physiological and pathophysiogical processes including neurodegeneration. In both HIV associated dementia in humans and its monkey model SIV encephalitis we find miR-21, a miRNA largely known for its link to oncogenesis, to be significantly upregulated in the brain. In situ hybridization of the diseased brain sections revealed induction of miR-21 in neurons. MiR-21 can be induced in neurons by prolonged N-methyl-D-aspartic acid receptor stimulation, an excitotoxic process active in HIV and other neurodegenerative diseases. Introduction of miR-21 into human neurons leads to pathological functional defects. Furthermore, we show that miR-21 specifically targets the mRNA of myocyte enhancer factor 2C (MEF2C), a transcription factor crucial for neuronal function, and reduces its expression. MEF2C is dramatically downregulated in neurons of HIV-associated dementia patients as well as monkeys with SIVE. Together, this study elucidates a novel role for miR-21 in the brain, not only as a potential signature of neurological disease but also as a crucial effector of HIV induced neuronal dysfunction and neurodegeneration.

Objective: To investigate morphologically the different changes of cornea tissue between thick and thin cornea flap with confocal microscope, and to compare visual function between thick and thin cornea flap after LASIK., Methods: 70 cases (70 eyes) who had received LASIK were divided into the thin cornea flap group (36 eyes, group A) and the thick cornea flap group (34 eyes, group B). The age was (23 +/- 5) years old, spherical equivalent diopter (SE) were ( - 4.76 +/- 2.30) D and (- 3.03 +/- 2.20) D. Inspection items included visual acuity (VA), refraction, wave-front analysis, contrast sensitivity analysis (KONTRASTOMETER BA4, Germany) during pre-and post-LASIK, at the same time, all eyes were examined by confocal microcopy (HRT III, Germany). Using Moria 90 microm and 130 microm keratome with Allegretto Excimer laser machine., Results: Refraction: there was no difference between the two groups. Post-LASIK 3 to 6 months: group A (0.35 +/- 0.21) D; group B (0.45 +/- 0.69) D (P > 0.05). Comparison results by confocal microscope. The thickness of cornea flap: group A (107.37 +/- 20.5) microm, group B (149 +/- 25.2) microm (P < 0.05). The thickness of wrinkle: group A (63.71 +/- 15.8) microm; group B (48.16 +/- 20.7) microm (P < 0.05). The thickness of acellular area: group A (69.93 +/- 15.8) microm; group B (55.63 +/- 23.7) microm (P < 0.05). The thickness of activation cornea cells: group A (60.15 +/- 30.9) microm; group B (51.86 +/- 27.9) microm (P < 0.05). The density of anterior stromal: group A (825.14 +/- 156.9) mm2; group B (853.54 +/- 126.8) mm2 (P < 0.05). There were significant differences on the thickness of cornea flap, the thickness of wrinkle, the thickness of acellular area, the thickness of activation cornea cells, the density of anterior stromal, but there was no difference on posterior stromal (P > 0.05). Wavefront analysis:there was no difference between two groups in Z3(-3), Z3(-1), Z3(1), Z3(3), Z4(0), RMS3, RMS4, RMS5, RMS6, RMSh (P > 0.05) pre-and post-operation. Contrast sensitivity analysis: there was also no difference between two groups (P > 0.05) pre-and post-LASIK 1-6 month., Conclusions: From the point of confocal microscope view, morphologic changes of cornea was significant with different thick flap. It is not cicatricial healing between the cornea flap and stromal. The cell injury of cornea tissue in the thin cornea flap was severe than that of in the thick cornea flap even though there was no significant difference between thin and thick cornea flap group in clinical refraction correction, wavefront and contrast sensitivity.

Objective: To investigate the efficacy, convenience and costs of recombinant follitropin alpha administered by a prefilled pen device and conventional syringe in Chinese women undergoing controlled ovarian stimulation for in vitro fertilization (IVF)., Methods: A total of 184 patients undergoing IVF treatment were enrolled in this study. According to a long-term recombinant follicle-stimulating hormone (rFSH) protocol, ovarian stimulation was performed with the prefilled pen and conventional syringe at random in these subjects, and the dose of follitropin, number of oocytes and embryo parameters and IVF-ET outcome were compared between the two groups., Results: The total rFSH dose, cost, and frequency of hospital visits were significantly lower in the pen protocol group, but the residual rFSH amount was higher. Compared with conventional injections, the prefilled pen was associated with significantly lowered rate of local redness, high rate of local bruise, more frequent follitropin dose modulation and lower serum oestradiol levels on HCG day. No significant difference was found in the endometrial thickness, numbers of oocytes retrieved, MII oocytes, transferred embryo, or the clinical pregnancy rates between the two groups. The ratio of MII oocytes, good quality embryo rates and implantation rates was significantly higher in the pen group with lower incidences of moderate and severe ovarian hyperstimulation syndrome., Conclusion: The prefilled pen provides an easy, safe, effective and more patient-friendly means for controlled ovarian stimulation procedure in Chinese women, but more attention should be given to protocol optimization and patient education.

The physiological activities of the cardiovascular system are under the control of autonomic nervous system (ANS). Recent researches have found that autonomic dysfunction, especially the withdrawal of vagal activity, was closely related to the etiology, course and prognosis of cardiovascular disease (CVD). Based on the current status and our achievements in this area, we discuss vagal regulation of different parts of the heart and the mechanism of vagal protection of myocardium. Using a force transducer and standard microelectrodes recording technology, we found that the vagus nerve transmitter--acetylcholine (ACh) had direct effects on ventricular myocytes in mammals: It inhibited the contractility and shortened the action potential duration of cardiac myocytes. We proved the existence of muscarinic receptors and vagal nerves innervation in ventricle with histochemical staining and molecular biological methods. Furthermore, ACh-activated potassium channel (KACh) was found in the ventricles of some animals by patch-clamp. Fade of the current (IK.ACh) to ACh in atrium was found in previous research, which was related to the muscarinic receptors and phosphorylation of G protein or potassium channel. However, the mechanism of the fade in ventricle needs to be further investigated. Combined with autonomic nervous evaluation methods (heart rate variability analysis) and relevant animal models, we studied the regulation of ANS during normal and morbid state, and proved the age-associated changes and compensatory effects of vagal control of hemodynamics after unilateral vagotomy. By increasing the vagal tension (ACh induced-preconditioning/postconditioning, aerobic exercise, beta receptor antagonist), we demonstrated protective effects of the vagus nerve on the ischemic myocardium and mechanism of the cholinergic anti-inflammatory effects on the inflammatory reaction induced by reperfusion injury. Evaluating cardiac autonomic nervous regulation and improving balance between sympathetic and vagal nerve will provide an important basis for the prevention and treatment of CVD.

Objective: The mechanisms of hypoxic-ischemic brain damage (HIBD) are still largely unknown. Elevation of intracellular calcium concentration and subsequent calcium-dependent proteases activation such as calpains seem to play an important role in the process of neuronal death. Calpain inhibitors showed neuroprotective effects in adult rat cerebral ischemia models. This study aimed to investigate the protective effect and associated mechanisms of calpain inhibitor-3 (MDL28170) on HIBD of neonatal rats., Methods: Seven-day old Sprague-Dawley rats were randomly divided into three groups: the control group (n = 18), HIBD group (n = 48) and calpain inhibitor-3 treated group (MDL group, n = 48). The mice in the latter two groups were subjected to hypoxia-ischemia (HI) insult. The puppies in MDL group were intraperitoneally injected with MDL28170 (25 mg/kg) at 0, 2 and 4 h after HI, while those in the other two groups were intraperitoneally injected with normal saline instead. All the pupies were sacrificed at 6 h, 24 h and 72 h after HI. Quantitative real-time fluorescent polymerase chain reaction was employed to detect micro-calpain gene expression, immunoblotting technique was used to measure mu-calpain and caspase-3 protein activation, apoptosis of ipsilateral cortex was detected by terminal deoxynucleotidyl transferase mediated d-UTP nick end labeling staining (TUNEL). CA1 neuronal loss was counted 24 h after HI by light microscopy., Results: After HI mu-calpain mRNA began to increase at 6 h and reached peak at 24 h compared to the control (1.805 and 4.83 vs. 1, P < 0.05); mu-calpain was activated through autolysis, the ratio of its activated fragment (76 000) vs. whole fragment (80 000) was significantly higher at 6 h (0.547 +/- 0.095) compared to the control (0.095 +/- 0.016, P < 0.05), it reached peak at 24 h (0.921 +/- 0.058, P < 0.01) and was still at a high level at 72 h (0.708 +/- 0.025, P < 0.05). Expression of activated caspase-3 protein reached peak at 24 h (3.78 +/- 0.30, P < 0.01), decreased to the same level as the control (1.56 +/- 0.07) at 72 h (1.82 +/- 0.11, P > 0.05). Apoptotic cells in the cortex ipsilateral to HI insult increased after HIBD, reached peak at 24 h (135.46 +/- 17.52/visual field) and was still markedly higher at 72 h (79.32 +/- 17.79/visual field) compared with the control (5.33 +/- 1.53/visual field, P < 0.01). At 24 h after HI CA1 neuronal loss (30.0 +/- 6.2/oil immersion lens field) in the HIBD group was significantly higher than that of the control (2.4 +/- 0.3/oil immersion lens field, P < 0.01). However, in the MDL group the expressions of mu-calpain and caspase-3 proteins were diminished, TUNEL positive cells at 6 h and 24 h were decreased and CA1 neuronal loss (18.2 +/- 2.4/oil immersion lens field, P < 0.05) was alleviated. The amount of micro-calpain mRNA was decreased in the MDL group, but there was no significant difference compared with the HIBD group., Conclusion: mu-calpain gene and protein expressions increased after HI, which may contribute to the pathogenensis of HIBD. Calpain inhibitor-3 may intervene neural necrosis and apoptosis by diminishing expressions of mu-calpain and caspase-3 to play a protective role after HI insult of neonatal brain.

Objective: To study optimal water extraction condition of LiYan granule., Methods: HPLC methods were developed for the determination of irisfloremin in LiYan granule. The contents of irisfloremin and extraction rate were selected as assessment indices. Three factors were studied by Doehlert design, including water-adding amount, decoction time and times of decoction., Results: The optimal extracting condition was water consumed as 8 times of curd herb, and 3 times for 2 hours each time., Conclusion: The water extraction process is feasible, and the experiment result can provide basic and instruction data for the ascertainment of water extraction process of LiYan granule.

Objective: To identify the expression of micro-calpain mRNA and protein in cerebral tissues of neonatal rats after hypoxic-ischemic brain damage (HIBD) and its role in the pathogenesis of HIBD., Methods: Seven-day-old Sprague-Dawley rats were randomly divided into seven groups: control group and groups with 0, 6, 12, 24, 48, and 72 hours of exposure to HIDB. Quantitative real-time fluroscent polymerase chain reaction and Western blot technique were employed to detect the expression of micro-calpain mRNA and its protein respectively., Results: The micro-calpain mRNA in the ipsilateral cerebral tissues began to increase 6 hours after the HIDB and reached peak at 24 hours, and then gradually decreased but remained in a higher level than that of the control group until at 72 hours (P < 0.05). The activated micro-calpain protein started to increase 6-12 hours after the HIDB (P < 0.05) and reached peak at 24 hours (P < 0.01). Higher levels of micro-calpain protein were maintained until 48 and 72 hours after the HIBD (P < 0.05)., Conclusion: The expressions of micro-calpain mRNA and protein increase with the HIBD. The micro-calpain may play a role in the pathogenesis of the HIBD.

Aim: To observe the correlation between vascular endothelial growth factor (VEGF) and CD147 expressed in the rheumatoid synovium and to investigate the effect of CD147 of cultured rheumatoid fibroblast-like synoviocytes (FLS) on the production of VEGF., Methods: The presence of CD147 and VEGF in the rheumatoid synovium derived from 15 patients with RA and 4 patients with osteoarthritis (OA) was detected by streptavidin/peroxidase (SP) immunostaining. FLS were cultured by enzymatic digestion of synovial tissues and incubated in 24-well plates. Then different concentration of LY294002, PD98059, SP600125, SB203580 and HAb18G mAb was added to each well. VEGF in the culture supernatant was measured by sandwich ELISA., Results: CD147 and VEGF in synovium from 15 patients with RA showed high expression, while CD147 and VEGF in synovium from 4 patients with OA showed low expression. Macrophages, fibroblast-like synovial cells and lymphocytes were demonstrated to express CD147 while synovial lining cells, fibroblasts surrounding microvessels and vascular smooth muscle cells were demonstrated to express VEGF. Statistic analysis indicates that VEGF production was correlated with the levels of CD147 expression. VEGFproduction was suppressed when CD147 expression was inhibited by LY294002 or HAb18G mAb., Conclusion: CD147 can regulate the angiogenesis in rheumatoid arthritis by VEGF. The low levels of CD147 expressed by FLS cells decrease the production of VEGF via the PI3K-Akt signaling pathway. These findings further highlight the importance of CD147 in pannus formation and angiogenesis.

Aim: To explore the percentages of CD4(+) CD25(high) regulatory T cells in peripheral blood or synovial fluid from patients with rheumatoid arthritis (RA) on different stages, and to study their correlations with the activity of the disease., Methods: Peripheral blood lymphocytes were obtained from the patients with active RA who had received no previous disease modifying (DMARDs) therapy(n=11), from individuals with stable, well-controlled RA (n=12), from subjects whose disease were poorly improved after DMARDs therapy(n=9), and from healthy controls(n=8). The frequencies of CD4(+) CD25(high) T cells were quantified by using flow cytometry(FCM). Meanwhile, the correlations between the percentage of CD4(+) CD25(high) T cells and the level of Anti-CCP antibody, CRP, ESR and RF were also investigated. Paired blood and synovial fluid was analysed in a small group of RA and other patients., Results: There was a smaller proportion of CD4(+) CD25(high) T cells in the peripheral blood of the active RA patients(mean 5.24%) and poorly-improved RA patients(mean 6.43%) than in patients with stable well-controlled RA or in healthy controls(mean 11.79% and 17.17%, respectively, P<0.01 in each case). The frequency of CD4(+) CD25(high) T cells from RA was negatively associated with Anti-CCP antibody(58.0 Ru/mL), ESR(38.8 mm/h) and CRP(2.73 mug/L), (P<0.05 for each). On contrast, The frequency of CD4(+) CD25(high) T cells from healthy controls was not significantly correlated with the level of Anti-CCP antibody(<5.0 Ru/mL), ESR(4.67 mm/h), CRP(0.15 mug/L) and RF(1.37) (all P>0.1). The percentage of CD4(+) CD25(high) regulatory T cells from synovial fluid of RA patients (24.32%) was significantly lower than that of AS patients(30.24%) (P<0.05)., Conclusion: The results demonstrate a smaller CD4(+) CD25(high) regulatory T cell population in peripheral blood of individuals with active RA prior to disease modifying treatment and with poorly-improved RA, and this is negatively associated with the activity of the disease.

Objective: To analyze the factors affecting the accuracy of Osaka formula multiparameter ultrasound-based fetal mass estimation, thereby establishing new formulas to improve the accuracy of the estimation., Methods: A retrospective review was conducted among 519 healthy women with singleton pregnancy. Three days before the delivery (between 37 and 42 weeks' gestation), ultrasonic measurement of the fetal weight and other indices of the fetus was routinely performed. Correlation and multiple linear stepwise regression analysis were used to correct the 3 equations, which, along with Osaka University formula, were used to predict another 219 fetuses' birth weight. The coincidence rate of the predicted value and with the actual birth weight, and the absolute error and relative error were compared between the equations., Results: The fetal abdominal area (AA) and abdominal circumference (AC) showed the most conspicuous influence on the estimated fetal birth weight, and fetal humerus length (HL) was more sensitive than femur length (FL) for the estimation. Three new regression equations were established, among which the equation 2 (fetal birth weight=1082.859+4.116xAAxHL) showed the best accuracy in clinical prediction., Conclusion: AA,AC and HL are more sensitive indices for estimation of the fetal birth weight, and the equation 2 established in this study still awaits further verification for its clinical value.

Objective: To study a simple and scientific method to produce the animal model of diabetic cardiomyopathy., Methods: Injecting the streptozotocin (STZ) liquid at the left iliac region only once in the model group, observing the general condition, the change of eating level, drinking level, urine level and weight, examining the level of creatine kinase ( CK- MB) , lactate dehydrogenase (LDH-1 ) , insulin, blood sugar and urine sugar. Observe the cardiac muscle ultrastructure, the large vessel tunica intima and electrocardiogram., Results: After injecting the STZ 3 weeks, the model rats appear morbid, the value of eating, drinking, urine are increased and weight decreased. The model rat blood sugar, urine sugar, CK and LDH levels are higher than control rat. The model rat insulin level is lower than control group. The model rat cardiac muscle ultrastructure accord with the diabetic cardiomyopathy' s, and the fibrous or atheromatous plaque do not been found at the large vessel tunica intima., Conclusion: The SD model rat appear the diabetic cardiomyopathy condition by injected the STZ (55mg/kg) at the left iliac region only once in 3 weeks. It is a simple and scientific method to produce the animal model of diabetic cardiomyopathy.

Objective: To investigate the general pattern of cholinergic nerve distribution and M(2) receptors in adult rat heart., Methods: Karnovsky-Roots histochemical staining combining point counting method and immunochemical SABC method with image analysis were used to identify the cholinergic nerves and M(2) receptors, respectively, in adult rat heart., Results: Positive staining of cholinergic nerves and M(2) receptors was found in all regions of the rat heart, and the point count of cholinergic nerves in the atria was 4.6 times as much as that in ventricles, and the area of immunoreactive substance for M(2) receptors two-fold higher in the atria than in the ventricles. The point counts of the cholinergic nerves in the medial-layer myocardium were fewer than that in subepicardial and endocardial tissues of the left ventricular free wall. However, M(2) receptors were comparable among the 3 layers of the left free ventricular wall., Conclusion: Cholinergic nerves and M(2) receptors are located in both rat atria and ventricles, but their density is much higher in the atria than in the ventricles. Transmural heterogeneity characterizes cholinergic nerve innervation in the left ventricular free wall without significant differences in M(2) receptor density.

The programmed cell death 5 (PDCD5) protein plays an important apoptosis-accelerating role in cells undergoing apoptosis. Decreased expression of PDCD5 has been detected in various human carcinomas. Here we describe that one potent short interfering RNA (siRNA) against the PDCD5 (siPDCD5) specifically inhibits the expression of PDCD5 at both the mRNA and protein level. Cells with decreased PDCD5 expression displayed reduced sensitivity to an apoptotic stimulus induced by Bax overexpression in HeLa, HEK293 and 293T cell lines. Furthermore, we also show that siPDCD5 inhibited both caspase-3 activity and procaspase-3 cleavage. Suppressed expression of PDCD5 attenuates the release of cytochrome c from mitochondria to cytosol induced by Bax overexpression. This phenomenon is accompanied by the reduced translocation of Bax from the cytosol to mitochondria. MTT assay shows that targeted suppression of PDCD5 expression markedly promoted cell proliferation in Hela and HEK293 cell lines. Our data suggests that PDCD5 may exert its effects through pathway of mitochondria by modulating Bax translocation, cytochrome c release and caspase 3 activation directly or indirectly, and that decreased PDCD5 expression may be one of the mechanisms by which tumor cells achieve resistance to apoptotic stimulus induced by anticancer drugs.

Autonomic nervous system plays an important role in the regulation of mammalian heart, and it is divided into the sympathetic and parasympathetic (vagal) subsystems. The parasympathetic (vagal) control of the atria involves modulation of chronotropic, dromotropic and inotropic activities, but the role of the parasympathetic innervation of the ventricles is still unclear. There is a common misconception that the sympathetic nerves innervate all over the heart; while the parasympathetic nerves only innervate the superventricular part of the heart, but not the ventricles. Recent evidence indicates that the cholinergic innervation of the left ventricle is functionally very important in some mammalian species. The present article reviews the evidence of vagal control in the ventricles from the anatomy and histochemistry, molecular biology, and function areas. Additionally we overview the vagal (muscarinic) regulation of cardiac contractile function and its signal transduction.

Conventional ischaemia-mimetic solutions contain several key components for inducing hypoxia, glucose deficiency, acidosis, lactate accumulation and hyperosmosis. The effect of each component on myocyte contractility during cardiac ischaemia was investigated in this study. A video-based edge-detection system was used to monitor single ventricular myocytes isolated from the rat. The effect of each factor was compared by preparing the following ischaemia-mimetic solutions: solution A, containing all of the above-mentioned factors; and solutions B, C, D, E and F, each with one of the factors excluded. The solutions that contained lactate severely reduced the contractility of the cardiomyocytes, but cell contraction did not differ significantly between the cardiomyocytes in these solutions. The effect of the solution without the acidosis-inducing component was weaker than that of the conventional ischaemia-mimetic solution. The solution lacking lactate produced the least depression of cell contractility. Lactate impaired cardiomyocyte contractility in a concentration-dependent manner. Our observations suggest that lactate is the main contributor to cardiac ischaemic injury and that its effects are attributable to acidosis and are concentration dependent. Imposition of hypoxia, glucose deficiency and hyperosmosis had little impact on the cardiomyocytes.

The article reviewed the research on the pharmacological effects of Danshen on hemorheology. We discussed the action mechanism of Danshen and its preparations from the points of hemorheology, blood and vessel endothelium cell. Danshen and its preparations can obviously improve the hemorheological characteristic by various ways.

The direct negative effects of acetylcholine (ACh) on guinea-pig atria and ventricles were investigated using standard microelectrodes, a force transducer and a video edge-detection system. It was found that: (1) ACh (at 0.001-100 microm) decreased the force of contraction and shortened the action potential duration (APD) in both atria and ventricles in a concentration-dependent manner, and that the atria were more sensitive to ACh than the ventricles; and (2) the direct negative inotropic effect of ACh (1 microm) on an isolated cardiac cell was similar to that on the isolated myocardium. But this effect was not present in all isolated ventricular cells, while all the atrial cells responded to ACh. In conclusion, ACh had direct inhibitory effects on both atrial and ventricular tissue and myocytes, although the effects were greater in atria than in ventricles; and the negative inotropic effect of ACh was closely related to the shortening of the APD.

Objective: To explore etiology, clinical manifestation and immunological changes of infectious pneumonia of neonates in Chengdu area., Methods: Serum specimens were collected from 111 infants with infectious pneumonia. Eight viral and mycoplasmal specific serum IgM antibodies were detected by enzyme linked immunosorbent assay (ELISA); C reactive protein (CRP), total IgG and its subclasses, IgA and IgM were determined by rate scattered nephelometry; T lymphocyte subpopulations were detected by biotin-streptavidin-peroxidase method, and clinical and other laboratory data were analyzed., Results: (1) Etiological agents: specific serum IgM antibodies were positive in 40 of 111 cases (36.0%) with pneumonias. All the 30 control infants were negative for the specific serum IgM antibodies. Among 111 infants with infectious pneumonia, 20.7% had single viral or mycoplasmal infection, 40.5% had bacterial infection, 15.3% had viral and mycoplasmal infection with bacterial infection; 23.4% had infection with unknown agents. (2) The most common clinical manifestations were tachypnea and cyanosis. The next were cough, milk choking, rales, retractions of the supraclavicular, intercostal and subcostal areas. Roentgenographic examination commonly revealed vague opacities, increased density and patchy infiltration. (3) Immune status: (1) CD(3), CD(4) cell counts of infants with pneumonias were lower than those of the controls while their serum IgA, IgM concentrations were higher than those of the control. (2) The CD(3) and CD(4) cell counts of the group with bacterial infection were lower than those of the control group. (3) The serum IgA concentration of the group with viral and mycoplasmal infection was higher than those of the control group and the group with unknown infection. (4) The serum IgM concentration of the group with bacterial infection was higher than those of the control group. (5) There were no significant differences in CD(8) cell counts, CD(4)/CD(8), concentration of serum IgG and IgG(1 - 4) between pneumonia group and the control group, and among various infectious groups and the control., Conclusion: Pathogens of neonatal infectious pneumonia in Chengdu area included single viral or mycoplasmic infection or bacterial infection, viral and mycoplasmal infection with bacterial infection, and unknown infection. Immunological changes of newborn infants suffered from infectious pneumonia included declined CD(3) and CD(4) cell counts, particularly in bacterial infection.

Objective: To explore and analyze the clinical manifestation, diagnosis, management and maternal-perinatal prognosis of severe acute respiratory syndrome (SARS)-complicated pregnancy in second and third trimesters., Methods: Clinical data of 5 inpatients with SARS-complicated pregnancy in second and third trimesters from 4(th) February to 17(th) March 2003 were analyzed retrospectively., Results: Five patients were all primigravida (including 2 twins). Two were infected in second trimester while the other 3 in third trimester with 2 hospital-acquired infections and 3 community-acquired infections. All patients had fever (5/5), 3 chills or rigor, 4 cough; 2 with decreased lymphocyte, 2 decreased platelet, 3 elevated alanine aminotransferase (ALT), 4 hypoalbuminemia, 5 abnormal chest radiographs. All 5 patients were cured with 1 requiring intensive care. 5 neonates including 1 twins have been followed up without evidence of SARS infections up to now. In a twin-pregnancy 1 fetus was lost while the pregnant's situation is stable., Conclusions: Common diagnostic criteria were Suitable for SARS-complicated pregnancy in second and third trimesters, but attention should be paid to the interaction between SARS and special pathological changes during pregnancy. Patients should be isolated and monitored intensively with timely cesarean section in severe cases, which could significantly decrease the maternal-perinatal mortality. The use of corticosteroids and psychological supports need further study.

The adherence and viability of central neural cells (substantia nigra) on a thin layer of SiO(2) on Si wafers with different surface roughness were investigated. Variable roughness of the Si wafer surface was achieved by etching. The nano-scale surface topography was evaluated by atomic force microscopy. The adherence and subsequent viability of the cells on the wafer were examined by scanning electron microscopy (SEM) and fluorescence immunostaining of tyrosine hydroxylase (TH). It is found that the surface roughness significantly affected cell adhesion and viability. Cells survived for over 5 days with normal morphology and expressed neuronal TH when grown on surfaces with an average roughness (Ra) ranging from 20 to 50 nm. However, cell adherence was adversely affected when surfaces with Ra less than 10 nm and rough surfaces with Ra above 70 nm were used as the substrate. Such a simple preparation procedure may provide a suitable interface surface for silicon-based devices and neurones or other living tissues.

The aim of this study was to investigate the cryobiological characteristics of placental cord blood (PCB) cryopereserved by using BioArchive auto-preserved liquid nitrogen system (BioArchive system). After Hespan depletion of red blood cells, 5 ml mixture of DMSO and 10% Dextran 40 were added into 20 ml of enriched leukocyte. 53 PCB units were cryopreserved as following protocol: pre-freeze rate 10 degrees C/min, start freeze temperature -3 degrees C, end freeze temperature -10 degrees C to -15 degrees C, post freeze rate 2 degrees C/min, and end temperature -50 degrees C. After rapid thawing at 38 degrees C, the PCB were washed with 5% human serum albumin -10% Dextran 40 and centrifuged at 400 x g, 10 degrees C for 20 minutes. The results showed that the viability of nucleated cells post-thaw was (73.3 +/- 12.5)%, the CD34(+) cell content was (0.3 +/- 0.21)% for pre-freeze PCB and (0.45 +/- 0.36)% for post-t haw PCB. No significant difference for CFU-GM/-G/-GEMM counts was found between pre-freeze and post-thaw PCB. Thawed PCB contained in two compartments (20 ml and 5 ml) of a freezing bag showed similar viability and clonogenic capacity. Differential count of white blood cell was significantly changed. For post-thaw PCB, it was dramatically decreased for the percentage of granulocytes, and highly increased for the percentage of lymphocytes and monocytes. It was concluded that the condition for cryopreservation and thawing of PCB may be harmful to mature cells, and cells with large size, such as granulocyte, but suitable to lymphocyte and monocyte, especially for the cells with small size, such as CD34(+) cells.