Polymorphisms of the Matrix Metalloproteinase Genes are Associated with Acute Ischemic Stroke in Chinese Han Population.

Background and Purpose: Studies have shown that matrix metalloproteinase (MMP-2,3,9) plays an important role in the pathologic process of ischemic stroke (IS). The aim of this study was to investigate the relationship between C1306T, 1612-5A/6A, C-1562T polymorphisms of MMP-2,3,9 genes and IS in Chinese Han population. Methods: The polymorphisms of MMP-2(C1306T), -3(1612-5A/6A), -9(C-1562T) gene were detected by PCR-RFLP and SNaPshot sequencing. Then, stratified analysis was used to study the relationship between IS subtypes and MMP-2,3,9 polymorphisms. Results: For the MMP-2 gene C1306T polymorphism, TT genotype and T allele were significantly associated with a reduced risk of IS (P = 0.015, P = 0.003, respectively). T allele was significantly associated with a reduced risk of small artery occlusion (SAO) subtype compared with the control group (P = 0.012, OR = 0.550, 95% CI = 0.065-1.291). For the MMP-3 gene-1612 (5A/6A) polymorphism, 5A/5A genotype was significantly increased in the IS group (P = 0.011, OR = 0.370, 95% CI = 0.168-0.814), especially in the large-artery atherosclerosis (LAA) subtype (P = 0.001, OR = 2.345) as compared to the control group. Conclusion: Our study suggested that the T allele of MMP-2 may be a protective factor of IS, especially in SAO subtype, while the 5A/5A gene of MMP-3 may increase the risk of IS, especially in LAA subtype in Chinese Han population.

The characteristic of nonmotor symptoms with different phenotypes and onsets in multiple system atrophy patients.

OBJECTIVES: The characteristic of nonmotor symptoms in patients with multiple system atrophy (MSA) has varied among previous studies. The objective was to investigatethe nonmotor characteristics in MSA patients with different phenotypes, sex and different onset patterns. METHODS: We performed a retrospective review of 1492 MSA patients. All cases were evaluatedby neurologists and assessed with motormanifestations, nonmotor symptoms, auxiliary examinationand brain MRI scans. RESULTS: Multiple system atrophy-cerebellar ataxia (MSA-C) was the predominant phenotype in 998 patients. Average age of onset (56.8 ± 9.2 years) was earlier, the disease duration (2.4 ± 2.2 year) was shorter and brain MRI abnormalities (49.2 %) were more frequently in MSA-C (P < 0.001). Multiple system atrophy-parkinsonism (MSA-P) patients were more likely to have nonmotor symptoms. After adjusted significant parameters, urinary dysfunction (OR 1.441, 95 %CI = 1.067-1.946, P = 0.017), constipation (OR 1.482, 95 %CI = 1.113-1.973, P = 0.007), cognitive impairment (OR 1.509, 95 %CI = 1.074-2.121, P = 0.018) and drooling (OR 2.095, 95 %CI = 1.248-3.518, P = 0.005) were associated with the MSA-P phenotype. Males were more likely to have orthostatic hypotension, urinary dysfunction, sexual dysfunction, drooling and females in constipation and probable RBD. In different onset patterns, constipation (59.2 %) and probable RBD (28.4 %) were more frequently in autonomiconset pattern. CONCLUSIONS: MSA-C is the predominant phenotype in Chinese patients, while many nonmotor symptoms are more common in MSA-P phenotype. Patients with different sex and onset patterns have different nonmotor characteristics. The different clinical features identified could help the physician counseling of MSA patients more easily and more accurately.

Effects of dietary supplementation with alpha-lipoic acid on apparent digestibility and serum metabolome alterations of sheep in summer.

To investigate the effects of alpha-lipoic acid (LA) on the nutrition metabolism of sheep (36.72 ± 1.44 kg) in the summer, twenty-one sheep were randomly assigned to three treatments addressing LA supplementation: 0.00 (CTL), 600 (LA-L), and 900 (LA-H) mg/kg of dry matter (DM) per day for each sheep. Whole feces and urine collection methods were used to analyze apparent digestibility; ELISA kits to determine the hormones, antioxidant, and immune parameters in the blood; and serum metabolomics to detect and analyze small molecular substances. The results showed the DM intakes in the LA-L and LA-H groups were significantly increased by 8.22% and 8.02%, respectively, compared to that in the CTL group, and there was no significant effect on average daily gain, feed conversion ratio, nitrogen digestibility, calcium digestibility, and phosphorus digestibility. Regarding hormones, antioxidant, and immune indicators, the concentrations of triiodothyronine, superoxide dismutase, glutathione reductase, HSP70, and IgA significantly increased after LA supplementation, while adrenaline and malondialdehyde levels significantly decreased. With the pairwise comparison of the three groups, metabolomics analysis identified 22 differential metabolites in the positive/negative modes, respectively, which suggested LA supplementation can significantly affect sheep's lipid, amino acid, and nucleic acid metabolism. Additionally, 3-indolepropionic acid, cinnamoylglycine, butyric acid, dodecanedioic acid, indoxyl sulfate, and pantothenic acid were the common differential metabolites with higher concentrations after LA supplementation. In summary, dietary supplementation of LA can increase the sheep's DMI, energy digestibility, antioxidant capacity, and immunity. These changes provide evidence to support the use of LA supplementation for livestock.

Metabolomic and Lipidomic Approaches to Evaluate the Effects of Eucommia ulmoides Leaves on Milk Quality and Biochemical Properties.

Eucommia ulmoides leaves (EUL) contain a variety of natural bioactive compounds including chlorogenic acid, geniposide acid, and aucubin. These bioactive chemicals improve immune function and regulate lipid metabolism. The aim of this study was to investigate the effects of EUL on the biochemical properties of milk. Twenty Holstein dairy cows were randomly allocated to two groups fed a control (CTR, diet without EUL, n = 10) or EUL (diet containing 3% EUL, dry matter, n = 10) diet for 55 d. At the end of the experimental period (d 55), milk samples were collected and analyzed to determine their composition. Though levels of milk fat, protein, lactose, and total milk solids were similar between the groups, small molecules, metabolites, lipids, and cytokines differed. Compared with the CTR group, the EUL group had an improved cluster of differentiation (CD)4/CD8 ratio (P < 0.05) and lower interleukin (IL)-8 and IL-6 content (P < 0.05). Metabolomics analysis identified 14 metabolites including 7Z, 10Z, 13Z, 16Z, 19Z-docosapentaenoic acid (FC = 3.129), adrenic acid (FC = 2.830), and eicosapentaenoic acid (FC=1.685) as having significantly increased in the EUL group (P < 0.05) while 11 metabolites, including indole-2-carboxylic acid (FC = 0.636), cholic acid (FC = 0.430), and creatine (FC = 0.784) had significantly decreased (P < 0.05). Based on a constructed metabolome map, linoleic acid metabolism had the highest impact value for EUL. A total of 21 lipid classes and 1,094 lipid species were detected in the milk by lipidomic analysis, among which 40 differed significantly between the CTR and EUL groups. The present findings showed that the EUL altered milk composition. Correlation analysis showed that 7Z, 10Z, 13Z, 16Z, 19Z-docosapentaenoic acid, adrenic acid, and eicosapentaenoic acid levels were negatively correlated with those of the inflammatory factors IL-6 and IL-8 (P < 0.05), indicating that EUL improved milk quality by reducing inflammatory factors and increasing the CD4/CD8 ratio. Overall, our data demonstrate that EUL had positive effects on milk antioxidant parameters, immune indices, and micro-composition metabolism, thereby improving milk quality.

Effect of melatonin on visceral fat deposition, lipid metabolism and hepatic lipo-metabolic gene expression in male rats.

Melatonin (MT) influences lipid metabolism in animals; however, the mechanistic effect of melatonin on liver fat and abdominal adipose deposition requires further clarity. In order to study the effects of melatonin on lipid metabolism, and hepatic fat and abdominal adipose deposition in animals, twenty Sprague-Dawley (SD) rats of 6 weeks of age with similar bodyweight were randomly divided into two groups: control (CTL) and MT-treated (10 mg/kg/day). During a 60-day experiment, food intake and bodyweight were measured daily and weekly respectively. At the end of treatment, blood samples were collected to collect plasma to quantify hormones and metabolic indicators of lipid metabolism. In addition, organ and abdominal adipose depots including liver, and omental, perirenal, and epididymal fat were weighed. Liver tissue was sampled for sectioning, long-chain fatty acid (LCFA) quantification, and gene chip and Real-time quantitative PCR (qPCR) analyses. The results showed that liver weight and index (ratio of liver weight to body weight) in MT group reduced by 20.69% and 9.63% respectively; omentum weight and index reduced by 59.88% and 54.93% respectively, and epididymal fat weight reduced by 45.34% (p = 0.049), relative to CTL. Plasma lipid indices, triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL) and total cholesterol (TC) with MT treatment decreased significantly compared with the control. Fat and 8 LCFA content in liver in MT group also decreased. Gene chip and qPCR demonstrated that there were 289 genes up-regulated and 293 genes down-regulated by MT. Further analysis found that the mRNA expression of lipolysis-related genes increased, while the mRNA expression of lipogenesis-related enzymes decreased (p < 0.05) with MT. This study concluded that melatonin greatly affected fat deposition, and hepatic LCFA supply and the expression of genes associated with lipogenesis and lipolysis.

Neural-specific distribution of transmembrane protein TMEM240 and formation of TMEM240-Body.

Mutation in TMEM240 is suggested to cause SCA21, but the specific mechanism has not been clarified. The subcellular localization, specific biological function, and corresponding mechanism of action of TMEM240 have also not been delineated. In this study, the mRNA and protein expression of TMEM240 were assessed using qPCR and western blotting, respectively. Live cell imaging was used to establish the sub-cellular location of TMEM240, and electron microscopy was used to determine the morphology and distribution of TMEM240 in the cell. TMEM240 was specifically expressed in the neurons. Exogenous TMEM240 formed a multilayered cell structure, which we refer to as TMEM240-Body (T240-Body). T240-Body was separated and purified by centrifugation and filtration. An anchor protein His-tagged-GFP-BP on Ni-NTA agarose was used to pull down T240-GFP binding proteins. Both the N-terminal and the C-terminal of TMEM240 were confirmed to be inside the T240-Body. Co-localization experiments suggested that peroxisomes might contribute to T240-Body formation, and the two transmembrane regions of TMEM240 appear to be essential for formation of the T240-Body. Emerin protein contributed to formation of T240-Body when combined with TMEM240. Overall, this study provides new insights into TMEM240, which inform future research to further our understanding of its biological function.

Administration of tissue plasminogen activator without coagulation results in a Chinese population.

Routine coagulation test before intravenous tissue plasminogen activator (tPA) use increases the door to needle time (DNT). We sought to evaluate the safety of tPA use without coagulation results and its impact on prognosis. In our stroke registry, tPA was delivered with coagulation results from December 2015 to April 2016 and without coagulation results from May 2016 to December 2016. Differences of demographics, clinical characteristic, and prognosis between these two groups were analyzed. In addition, logistic regression analysis was conducted to identify predictors for DNT of over 60 min. A total of 201 stroke patients were included in the final analysis. Of these, 81 patients received tPA with coagulation results and 120 patients without coagulation results. Only one (0.8%) patient with abnormal coagulation results met the exclusion criteria of tPA use in patients without coagulation results. The difference of DNT between groups with (mean, 61.7 min) and without (mean, 41.9 min) coagulation results was significant (P = 0.00). The group without coagulation results had a higher rate of favorable 90-day outcome (74.2 vs 70.4%) and lower rates of symptomatic intracranial hemorrhage/nonintracranial hemorrhage (4.9 and 22.2% vs 1.7 and 19.2%) than the group with coagulation results did; these differences were not statistically significant. In multivariate analysis, only tPA use with coagulation results was the predictor for DNT of over 60 min (P = 0.0030, OR = 2.44, 95% CI 1.28-4.65). The present study suggests that tPA could be delivered safely without coagulation results in patients without suspected coagulopathy, and avoiding coagulation tests reduces significantly the DNT interval.

Validation of simplified FABS scale to predict stroke mimics in a Chinese population undergoing intravenous thrombolysis.

OBJECTIVES: A large number of suspected stroke patients undergoing intravenous thrombolysis are stroke mimics (SMs). In this study, we sought to revise the FABS scale for screening and stratifying SMs from acute ischemic stroke (AIS) in a Chinese stroke population receiving fibrinolytic therapy. PATIENTS AND METHODS: The simplified FABS (sFABS) scale includes 4 items with 1 point for each item present: absence of facial droop, negative history of atrial fibrillation, age <50years, systolic blood pressure <150mm Hg at presentation. We evaluated consecutive suspected stroke patients undergoing intravenous thrombolysis in our stroke center for validation of sFABS scale. Diagnosis of SMs was based on absence of acute ischemic lesions on first and second diffusion weight imaging sequence in addition to an alternate diagnosis at discharge. RESULTS: A total of 190 AIS patients and 28 SMs were included in this study from December 2015 to February 2017. The sFABS scale showed excellent discrimination (C statistic: 0.928, 95% CI: 0.887-0.969, P<0.001). The Hosmer and Lemeshow goodness of fit test showed that the sFABS scale also had a good calibration (Cox and Snell R2=0.294, Nagelkerke R2=0.549). The plot of observed versus predicted risk of SMs showed high correlation (Pearson correlation coefficient: 0.983) between observed and predicted risk in our registered stroke population. CONCLUSION: The sFABS scale had excellent discrimination and good calibration abilities to predict SMs among a Chinese stroke population receiving tPA therapy. Further imaging evaluation may be necessary before the use of tPA if the sFABS score is higher.

Minor Nonintracranial Hemorrhage and Poor Prognosis among Stroke Patients Undergoing Intravenous Thrombolysis.

BACKGROUND: Whether nonintracranial hemorrhage (NICH) associated with intravenous thrombolysis (IVT) is a predictor of intracranial hemorrhage (ICH) and poor prognosis is ambiguous. We sought to analyze the rate of NICH and the relationship between NICH and poor outcome in the ischemic stroke population undergoing IVT. METHODS: This is a single-center, hospital-based prospective study. All ischemic stroke patients undergoing IVT between December 2015 and November 2016 were included. NICH was defined according to the criteria of the Bleeding Academic Research Consortium (BARC). ICH associated with IVT was defined based on the European Cooperative Acute Stroke Study II definition. On the basis of the modified Rankin Scale (mRS), 90-day outcome was divided into favorable outcome (mRS score 0-1) versus unfavorable outcome (mRS score 2-6) and independency (mRS score 0-2) versus dependency and death (mRS score 3-6). RESULTS: A total of 212 patients undergoing IVT were included in the analysis. Forty-five NICH events were reported in 42 patients (19.8%). Older age was independently associated with NICH (P = .049, odds ratio [OR] = .97, 95% confidence interval [CI] .94-1.0). Neither NICH with BARC class 1 or higher (P = .56, OR = .61, 95% CI .11-3.24) nor NICH with BARC class 2 or higher (P = .87, OR = 1.19, 95% CI .14-10.23) was associated with ICH. NICH with BARC class 1 or higher was not associated with unfavorable outcome (P = .67, OR = 1.17, 95% CI .56-2.45) and dependence and death (P = .47, OR = .72, 95% CI .30-1.75), neither was NICH with BARC class 2 or higher (P = .97, OR = 1.02, 95% CI .46-2.27 and P = .30, OR = .59, 95% CI .22-1.62). CONCLUSIONS: NICH was common among ischemic stroke populations receiving IVT. NICH with BARC class 2 or lower was not associated with ICH and poor outcome.