The Combination of Homocysteine and C-Reactive Protein Predicts the Outcomes of Chinese Patients with Parkinson's Disease and Vascular Parkinsonism

BACKGROUND: The elevation of plasma homocysteine (Hcy) and C-reactive protein (CRP) has been correlated to an increased risk of Parkinson's disease (PD) or vascular diseases. The association and clinical relevance of a combined assessment of Hcy and CRP levels in patients with PD and vascular parkinsonism (VP) are unknown. METHODOLOGY/PRINCIPAL FINDINGS: We performed a cross-sectional study of 88 Chinese patients with PD and VP using a clinical interview and the measurement of plasma Hcy and CRP to determine if Hcy and CRP levels in patients may predict the outcomes of the motor status, non-motor symptoms (NMS), disease severity, and cognitive declines. Each patient's NMS, cognitive deficit, disease severity, and motor status were assessed by the Nonmotor Symptoms Scale (NMSS), Mini-Mental State Examination (MMSE), the modified Hoehn and Yahr staging scale (H&Y), and the unified Parkinson's disease rating scale part III (UPDRS III), respectively. We found that 100% of patients with PD and VP presented with NMS. The UPDRS III significantly correlated with CRP (P = 0.011) and NMSS (P = 0.042) in PD patients. The H&Y was also correlated with Hcy (P = 0.002), CRP (P = 0.000), and NMSS (P = 0.023) in PD patients. In VP patients, the UPDRS III and H&Y were not significantly associated with NMSS, Hcy, CRP, or MMSE. Strong correlations were observed between Hcy and NMSS as well as between CRP and NMSS in PD and VP. CONCLUSIONS/SIGNIFICANCE: Our findings support the hypothesis that Hcy and CRP play important roles in the pathogenesis of PD. The combination of Hcy and CRP may be used to assess the progression of PD and VP. Whether or not anti-inflammatory medication could be used in the management of PD and VP will produce an interesting topic for further research

Potential role of mesenchymal stem cells in alleviating intestinal ischemia/reperfusion impairment.

BACKGROUND: Transplantation of bone marrow mesenchymal stem cells (MSCs) provides a promising therapeutic efficiency for a variety of disorders caused by ischemia or reperfusion impairment. We have previously demonstrated the efficacy of MSCs in mitigating intestinal ischemia/reperfusion (I/R) injuries in rats, but the mechanism by which MSCs engraft ameliorates I/R injuries has largely been unknown. The present study aimed at investigating probable mechanisms by which MSCs exert their function. METHODS: Male donor derived rat MSCs were implanted into intestine of female recipient rat by direct submucosal injection after superior mesenteric artery clamping and unclamping. The homed MSCs were detected by Y chromosome in situ hybridization probe, and the tumor necrosis factor-α (TNF-α) content in intestinal mucosa was determined by ELISA. Expression of proliferative cell nuclear antigen (PCNA) in bowel mucosa was assayed by real-time PCR and intestinal mucosa expression of phosphorylation extracellular signal-regulated kinase (pERK1/2) and nuclear factor-κB (NF-κB) were evaluated by western blot. RESULTS: Four and seven days after MSCs transplantation, the TNF-α content of bowel mucosa in MSCs group was significantly lower than that in saline group. The PCNA in bowel mucosa showed higher expression in MSCs treated group than the saline group, both at 4 and 7 days after cell transplantation. The expression of intestinal mucosal pERK1/2 in MSCs treated group was markedly higher than that in saline group, and the expression of NF-κB in MSCs treated group was noticeably decreased than that in saline group at 4 and 7 days post MSCs transplantation. CONCLUSION: The present investigation provides novel evidence that MSCs have the potential to reduce intestinal I/R injuries probably due to their ability to accelerate cell proliferation and decrease the inflammatory response within intestinal mucosa after ischemia and reperfusion

Expression of pERK1/2 in intestinal mucosa at 4 and 7 days after MSCs transplantation.

<p>(<b>A</b>) Bands of the expression of pERK1/2 and its own tERk1/2 protein were detected by western blot. (<b>B</b>) The ratio value of pERK1/2 to its own tERK1/2 of the bands which was evaluated densitometrically using the software Quantity One for the three groups at different time points. Each bar represents mean ± SD of the ratio value in every group. S: sham group; C: control (saline) group; M: MSCs group. 4d and 7d represent 4 days and 7 days postoperatively. *<i>: P<0.05</i>, compared with control (saline) group, by one-way ANOVA test.</p

MSCs homing photograph (40×).

<p>After cell transplantation, engrafted cells were detected by Y chromosome <i>in </i><i>situ</i> hybridization at 1, 4, 7, and 10 days (the green fluorenscent point represents positive donor derived cells), which corresponded to graphs A, B, C, and D respectively. Most of the homed cells were located at mucosal lamina propria, and the amounts of engrafted cells were more at 4 and 7days than 1 and 10 days postoperatively.</p

Mucosal NF-κB expression in each group at 4 and 7 days after MSCs administered.

<p>(<b>A</b>) Semiquantitative assessment of bands for NF-κB and its internal control protein. (<b>B</b>) Values of the NF-κB band performed densitometrically using the software Quantity One. Each bar represents the mean ± SD of the different groups; values were compared by one-way ANOVA test, <i>**: P<0.01</i>, <i>*: P<0.05</i>. S: sham group; C: control (saline) group; M: MSCs group. 4d and 7d represent 4 days and 7 days postoperatively.</p

ELISA analysis of intestinal mucosal TNF-α content among the three groups.

<p>Data are shown as means ± SD of the three groups, <i>*P < 0.01</i> vs. MSC group; # <i>P <0.05</i> vs. Sham group, values are evaluated by one-way ANOVA test.</p

Real-Time PCR analysis of PCNA expression in the intestinal mucosa.

<p>Data are means ± SD of each group at different time point and expressed as the ratio of original value mRNA of PCNA to its internal control mRNA of GAPDH. S: sham group; C: control (saline) group; M: MSCs group. 4d and 7d represent 4 days and 7 days postoperatively. ** <i>P<0.01</i>, <i>* P<0.05</i>, compared with control (saline) group, by one-way ANOVA test.</p

SH2 Superbinder Modified Monolithic Capillary Column for the Sensitive Analysis of Protein Tyrosine Phosphorylation

In this study, we present a method to specifically capture phosphotyrosine (pTyr) peptides from minute amount of sample for the sensitive analysis of protein tyrosine phosphorylation. We immobilized SH2 superbinder on a monolithic capillary column to construct a microreactor to enrich pTyr peptides. It was found that the synthetic pTyr peptide could be specifically enriched by the microreactor from the peptide mixture prepared by spiking of the synthetic pTyr peptide into the tryptic digests of α-casein and β-casein with molar ratios of 1:1000:1000. The microreactor was further applied to enrich pTyr peptides from pervanadate-treated HeLa cell digests for phosphoproteomics analysis, which resulted in the identification of 796 unique pTyr sites. In contrast, the conventional SH2 superbinder-based method identified 41 pTyr sites for the same sample, only 5.2% of the number achieved by the microreactor. Finally, this microreactor was also applied to analyze the pTyr in Shc1 complex, an immunopurified protein complex, which resulted in the identification of 15 pTyr sites. Together, this technique is best fitted to analyze the pTyr in minute amount of sample and will have broad application in fields where only a limited amount of sample is available

Comparison of age, BMI, Hcy, CRP, UPDRS (III), MMSE, and NMSS between PD and VP.

<p>* <i>P</i><0.01.</p><p>Abbreviations: SD, Standard deviation; BMI, Body Mass Index; MMSE, mini-mental state examination; UPDRS III, the unified Parkinson's disease rating scale part III; and NMSS, non-motor symptoms scale for Parkinson's disease (range of possible scores from 0 to 360).</p