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Objective:To explore the association of inflammatory markers and thromboinflammatory factors before and after thrombolytic intervention with functional outcomes in elderly patients with acute cerebral infarction.Methods:392 patients with acute cerebral infarction admitted to our hospital were randomly selected as study subjects and divided into an observation group(196 cases)treated with arterial thrombolytic therapy and a control group(196 cases)treated with intravenous thrombolysis.Functional outcomes of patients were assessed 72 hours after thrombolysis using the activities of daily living(ADL)scale and, based on the results, patients were divided into a poor functional outcome group and a good functional outcome group.Inflammatory markers such as neutrophil-to-lymphocyte ratio(NLR)and platelet-to-lymphocyte ratio(PLR)and thromboinflammatory factors such as monocyte chemoattractant protein-1(MCP-1), tissue plasminogen activator(t-PA), soluble CD40 ligand(sCD40L)and P-selectin before and after thrombolysis were measured.The relationship of these inflammatory markers and thromboinflammatory factors before thrombolysis with functional outcomes 72 hours after thrombolysis was analyzed.Results:NLR and PLR levels in the two groups after thrombolysis were significantly lower than those before thrombolysis(all P<0.05); Their levels in the observation group were lower than those in the control group(all P<0.05). After thrombolysis, MCP-1 levels in both groups were significantly higher and t-PA, sCD40L, P-selectin levels were significantly lower than pre-thrombolysis levels(all P<0.05); After thrombolysis, the observation group had better results than the control group(all P<0.05). Correlation analysis showed that NLR, PLR, MCP-1 and t-PA were positively correlated with NIHSS score( r=0.336, 0.264, 0.483, 0.549, all P<0.05). NLR, PLR, MCP-1, t-PA and sCD40L levels were significantly lower and P-selectin levels were significantly higher in the good functional outcome group than in the poor functional outcome group both before and 72 hours after thrombolysis( t=13.850, 18.208, 23.636, 22.371, 59.868, 96.646, 378.112, 141.213, 131.160, 110.039, 10.716, 11.108, P<0.05 for all). Logistic regression analysis showed that abnormal levels of NLR, PLR, MCP-1 and t-PA before and after thrombolysis were risk factors for adverse outcomes with thrombolytic intervention( P<0.05). ROC curves showed that the levels of NLR, PLR, MCP-1, t-PA, sCD40L and P-selectin before thrombolysis had a certain predictive value for the risk of adverse functional outcomes with thrombolysis. Conclusions:The levels of these inflammatory markers and thromboinflammatory factors before and after thrombolytic intervention have varying degrees of correlation with functional outcomes in elderly patients with acute cerebral infarction.
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Objective:To explore the quality of inventory samples of a biobank stored in a deep freezer from 0 to over 10 years in Shanghai Ruijin Hospital. Methods:We extracted 24 pairs of stocked gastric cancer samples between 2003 and 2014. We used 1%aga-rose gel electrophoresis to analyze DNA and RNA purity and integrity while adding the RNA integrity number (RIN) for precise analysis. Bicinchonininc acid (BCA) assay was used for protein concentration evaluation. Coomassie brilliant blue method was used for protein integrity assay. Results: The samples were divided into four groups according to cryopreservation period (9 years). No significant difference in DNA integrity was found between the groups (P>0.05);however, DNA degradation in normal gastric mucosa was faster than that in gastric cancer tissue (P=0.023). The RIN significantly declined when the storage period was 6 years or longer (P=0.018). No significant difference in protein concentration was observed between different groups. Using Coo-massie brilliant blue method, we found significant differences in preserved proteins with different molecular weights. Proteins with varying molecular weights were detected in the groups with the following cryopreservation periods:>9 years, a small number of low-molecular-weight (average 36.5 KD) proteins;6-8 years, medium-molecular-weight (average 65.63KD) proteins;3-5 years, high-molecu-lar-weight (average 127.5 KD) proteins;<2 years, high-molecular-weight (average 160 KD) proteins. Conclusion:Cryopreservation does not exert an obvious effect on DNA. If the cryopreservation period is more than 5 years, serious degradation of RNA should occur;like-wise, degradation of proteins with higher molecular weight should occur.
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BACKGROUND:For treatment of central nervous system diseases, neural stem cel s (NSCs) or bone marrow stromal stem cel s (BMSCs) can be transplanted into the brain, but there are less reports to compare the effects of two kinds of stem cel transplantation. OBJECTIVE:To explore the effect of midbrain NSCs and BMSCs on the behavior and brain morphology of rats with Parkinson’s disease. METHODS:Fifty-eight Sprague-Dawley rats were enrol ed to establish Parkinson’s disease models, and then randomly divided into three groups, which were treated with 5μL midbrain NSCs (n=20), 5μL BMSCs (n=20) and 5μL normal saline (n=18) via two coordinate points of the right striatum at 3 weeks after modeling, respectively. At 5 months after transplantation, the rats underwent intraperitoneal injection of apomorphine to observe behavioral changes, and then, the striatum was taken for immunohistochemistry staining. RESULTS AND CONCLUSION:The number of rotations was reduced significantly in the BMSCs and midbrain NSCs groups at 5 months after transplantation (P0.05). In the BMSCs group, BrdU/Nestin positive cel s were seen in the brain stratium at 1 week after transplantation;BrdU/GFAP and BrdU/NSE positive cel s as wel as TH positive cel s rather than BrdU/TH positive cel s were found in the brain stratium at 1 month after transplantation;after that, the number of BrdU/Nestin positive cel s was reduced gradual y and disappeared ultimately, but there were stil a certain number of BrdU/GFAP and BrdU/NSE positive cel s, especial y the former ones. Meanwhile, the NSCs group also had a similar situation, but no double-labeled cel s were in the normal saline group. These findings indicate that midbrain NSCs and BMSCs transplantation can both improve the behavior of Parkinson’s disease rats, and differentiate into neurons, astrocytes and dopaminergic neurons.
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Objective To compare the effects of telmisartan and (or) amlodipine on the reversal left ventricular re-modeling in two-kidney one clip hypertensive rats. Methods A total of 50 healthy male SD rats were randomly divided into 5 groups (n=10):two-kidney one clip renal hypertensive (2KIC) model group, sham group, telmisartan (10 mg/kg) group, am-lodipine (2.5 mg/kg) group and telmisartan (10 mg/kg)+amlodipine(2.5 mg/kg) group. The model of two-kidney one clip re-nal hypertensive rats was established. The tail arterial blood pressure was detected once a week. After 20 weeks, rats were sacrificed and specimens were collected. The left ventricular mass index (LVMI) was assessed. The myocardial ultrastructur-al changes were observed by electron microscope. Values of plasma renin activity (PRA), angiotensionⅡ(AngⅡ) and atrial natriuretic peptide (ANP) were measured by enzyme linked immunosorbent assay (ELISA).Results Compared with sham group, the levels of systolic blood pressure (SBP), LVMI, PRA, AngⅡand ANP were significantly higher in 2KIC group (P＜0.01). Compared with 2KIC group, values of SBP, LVMI, PRA and ANP were significantly lower in telmisartan group and am-lodipine group (P＜0.01), but the value of AngⅡwas significantly higher (P＜0.01). The levels of SBP, LVMI, AngⅡand ANP were significantly lower in combined medication group than those of single drug medication group (P＜0.01). There was no significant difference in the plasma PRA level between those groups (P>0.05). Results of myocardial electron microsco-py showed that the left ventricular remodeling was significantly improved in combined treatment group. Conclusion Telmisartan and amlodipine can effectively improve the left ventricular remodeling induced by hypertension. There was more effective therapy using both medications together.
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In this study, about 350 bp cDNA fragment was amplified by PCR. After being sequenced, the AE1-c-end gene fragment was cloned into EcoR I-Pst I site of pGADT7 to form AD ends in the yeast two-hybrid system. The recombinant plasmid was transformed into yeast AH109, and the expression in the yeast was observed. The results demonstrate that AE1-c-end was obtained. pGADT7-AE1-c-end has no toxic effect on the yeast. It can serve as a target gene of yeast two-hybrid system.
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Clonagem Molecular , DNA Complementar , Plasmídeos , Reação em Cadeia da Polimerase , Técnicas do Sistema de Duplo-Híbrido , LevedurasRESUMO
AIM: To obtain the glycopohorin A (GPA) cDNA and construct the target gene in yeast two-hybrid.METHODS: About 410 bp cDNA fragment was amplified from K562 cell by RT-PCR.After being sequenced, the GPA gene fragment was cloned into EcoR -Ⅰ- Pst Ⅰ site of pbridge to form BD ends in yeast two-hybrid system. The recombinant plasmid was transfered into yeast AH109, and the expression in the yeast was also examined. RESULTS: The amino acid sequence encoded by cloned cDNA was mostly the same as reported GPA, and about 1 mm white yeast clone grew in the selective medium after 3 d.CONCLUSION: pbridge-GPA has nontoxic to the yeast, which can serve as a target gene in yeast two-hybrid system.
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Acute alveolar injury was induced in rats by subcutaneous injection of Nnitroso-N-methylurethane (NMU) 6mg/kg body wt. NMU decreased the content of surfactent phosphoLipids, and increased the protein of alveolar lavage return of rats after 2 days of the injection. Such changes were not observed in lamellar bodies and microsomcs. Composition of phospholipids also changed in all lung fractions it showed that the ratio of PG/PI was decreased significantly in response to NMU. The molecular species profiles of both acidic phospholipids in the lung fractions were distinctly different from each other and the main molecular species were attered during the acute alveolar injury.
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AIM: To study the effect of the overexpression of p16 on an anion exchange function of band 3 in HeLa cells. METHODS: The expression of p16 and band 3 in HeLa cells was detected by immunohistochemistry (IHC). The p16 cDNA was subcloned to plasmids pEGFP-C1 by PCR and identified by restriction enzyme digestion and sequencing, and then, the recombinant pEGFP-C1-p16 plasmids were transiently transfected into HeLa cells. The expression of fusion protein in HeLa cells was detected by fluorescence microscope. 6-methoxy-N-(3-sulfopropyl)-quinolinium(SPQ)fluorescent probes were used to detect the anion exchange function of band 3. RESULTS: P16 and band 3 were expressed in HeLa cells. The amplificated p16 cDNA sequence was the same as the report sequence. The transfective efficacy of pEGFP-C1-p16 was above 60%. The anion exchange function increased after the transfection of pEGFP-C1-p16 plasmids. CONCLUSION: p16 facilitates the anion exchange function of band 3 in HeLa cells.