In the present study, packaging system composed of pAAV-CMV-GFP, pAAV-RC and pHelper were transfected into human embryonic kidney 293 cells (HEK293 cells) mediated by polyethyleneimine (PEI) to explore an optimal transfection condition. Different total plasmid DNA dosages (1, 2, 3, 4, 5, 6 μg) and different PEI/Plasmid ratios (1:1, 3:1, 5:1, 7:1) were tested with detection of green fluorescence protein (GFP) with ImagePro Plus6. 0 Software. Then transfection efficiency of the optimized transfection system was further observed for different time periods(12, 24, 36, 48, 60, 72 h). The results showed that total plasmid dosage of 4 μg/well with PEI/plasmid ratio of 3 : 1-5 : 1 was an efficient transfection condition. Transfection efficiency-time curve was an S-shaped curve. Transfection efficiency reached a plateau at 60 h after transfection. The optimized conditions for PEI-mediated transfection at the optimal time result in enhanced transfection efficiency of triple plasmid into HEK293 cells.
Green Fluorescent Proteins ; HEK293 Cells ; Humans ; Plasmids ; Polyethyleneimine ; Transfection ; methods

Dryness is the inherent performance in traditional Chinese medicine. Dryness with a specific efficacy and side effect can be reduced suitably by processing and compatibility in the clinical application. Nowadays domestic scholars have developed research of dryness in traditional Chinese medicine. However, it remains problems such as evaluation index of dryness not clear. This paper takes medical literature mining technology to analyze the historical origin and features of dryness theory. Combing the modern literatures to explicate the dryness' research status and existing problems. Putting forward the traditional Chinese medicine and research should adopt multidisciplinary knowledge and study the system of comprehensive evaluation. Dryness is expected to further application in traditional Chinese medicine clinical research.
Data Mining ; Drug Compounding ; methods ; Humans ; Medicine, Chinese Traditional ; Research

Objective To evaluate the assessment of tumor's size with ultrasound in research of prognosis of liver transplantation for hepatocellular carcinoma (HCC). Methods Clinical data of 148 patients with HCC who underwent liver transplantation were analyzed retrospectively. Results One-, 2-,3-,and 5-year overall actuarial survival were 73.3% ,45.6% ,35.4% ,and 32.1%,respectively. One-,2-,3-,and 5-year overall recurrence-free survival were 70.7 %, 44.3 %, 38. 5%, and 34. 5%, respectively. The overall tumor recurrence rate was 43.2%. Univariate analysis indicates that (the Kaplan-Meier method with the Log-Rank test) the total tumor burden (TTB) (χ2=15.098,P=0.001) was found to be significantly affecting the actuarial survival. While TTB (χ2=29. 038, P<0.001) was for recurrence-free survival. In multivariate analysis (with the multivariate Cox proportional hazards model), TTB (R2=1.610, P =0. 008) was found to be an independent predictor of actuarial survival. On the other hand, TTB (R2 =2. 206, P<0.001) was identified as the prognostic factor independently related to recurrence-free survival. Conclusions TTB is an independent prognostic factor for patients with liver transplantation for HCC. Assessment of tumor size with ultrasound is beneficial to the evaluation of indication for liver transplantation when patients with HCC were concerned.

A biosurfactant-producing strain(S_6)was isolated from oil-containing wastewater in oxidation ditch and identified as Pseudomonas aeruginosa based on physiological and biochemical experiments and 16S rDNA sequence analysis.Infrared spectrum analysis revealed that S_6 produced glucolipid in the process of metabolism.It was observed that S_6 decreased the surface tension of water from 72 mN/m to 33.9 mN/m with the critical micelle concentration(CMC)of 50mg/L.The measurement of oil displacement and surface tension demonstrated that the fermented liquid had stable surface activity at varying range of salinity,pH,amount of dissolved oxygen.The optimal culture condition was obtained through orthogonal experiment:glucose 10g/L,urea 5g/L,KH_2PO_4 1g/L,liquor of microelement 2mL,pH 8.0,water 1000mL;and the biosurfactant production under optimal culture condition was 0.173g/L.

Objective: To compare the safety and efifcacy of platelet glycoprotein IIb/IIIa antagonist in treating STEMI patients by intracoronary-intravenous administration and intravenous administration.
Methods: We searched PubMed, Embase, Cochrane library, CNKI, VIPH and Wanfang database, the retrieval stopped at 2014-03. According to 5.0.2 Cochrane handbook, 2 scientists collected 2494 STEMI patients treated by IIb/IIIa antagonist from 20 references, and they were divided into 2 groups. Combination group, the patients received intracoronary, then intravenous administration, n=1258 and Intravenous group, the patients receive only intravenous administration, n=1236. RevMan 5.0 software was used for Meta-analysis.
Results: At 1 month after PCI treatment, compared with Intravenous group, the Combination group had better conditions of TIMI 3, TMP 3, ST segment recovery, MACE occurrence and MI area changes, all P<0.01; Combination group also showed better conditions of angina recurrence, death and post-operative target vessel revascularization, all P<0.05. LVEF was similar between 2 groups at 1 week after PCI. MI recurrence, post-operative bleeding and thrombocytopenia were similar between 2 groups at 1 month after PCI, all P>0.05.
Conclusion: Intracoronary-intravenous administration of platelet glycoprotein IIb/IIIa antagonist had the better effect for treating STEMI patients without increasing the side effects of post-operative bleeding and thrombocytopenia.

To observe a PPAR-alpha agonist effect of N-oleoylethanolamine (OEA) on CB2 (cannabinoid receptor 2), an anti-inflammatory receptor in vascular endothelial cell, healthy HUVECs and TNF-alpha induced HUVECs were used to establish a human vascular endothelial cell inflammatory model. Different doses of OEA (10, 50 and 100 micromol x L(-1)) had been given to HUVECs, cultured at 37 degrees C for 7 h and then collected the total protein and total mRNA. CB2 protein expression was detected by Western blotting and CB2 mRNA expression was assayed by real-time PCR. As the results shown, OEA (10 and 50 micromol x L(-1)) could induce the CB2 protein and mRNA expression, but not 100 micromol x L(-1). To detect if anti-inflammation effect of OEA is partly through CB2, CB2 inhibitor AM630 was used to inhibit HUVEC CB2 expression, then the VCAM-1 expression induced by TNF-alpha was detected, or THP-1 adhere to TNF-alpha induced HUVECs was examined. OEA (50 micromol x L(-1)) could inhibit TNF-alpha induced VCAM-1 expression and THP-1 adhere to HUVECs, these effects could be partly inhibited by a CB2 inhibitor AM630. The anti-inflammation effect of OEA is induced by PPAR-alpha and CB2, suggesting that CB2 signaling could be a target for anti-atherosclerosis, OEA have wide effect in anti-inflammation, it may have better therapeutic potential in anti-inflammation in HUVECs, thus achieving anti-atherosclerosis effect.

Objective To investigate clinical features and the pathophysiology of 43 children with atypical idiopathic partial epilepsies of childhood(IPEC)that is unclassified according to the International League Against Epilepsy classification.Methods All the children with partial epilepsy of childhood in out hospital were followed up,including those age-related cases with benign process and excluding those with benign epilepsy of childhood with centro-temporal EEG foci,Panayiotopoulos type and Gastaut type.We reviewed their EEGs,seizures and therapeutic data to analyze the semiologieal characters and the interietal activities during they were awake and sleeping.Results The average age of onset of epilepsy was 8.84years old.Frontal absences(43.2%),adversive seizures(47.7%)were more common.Everyday seizures occurred in 38.6%of patients and monthly seizures in 56.8%of patients.Atypical focal(43.2%),multifocal(27.3%),and generalized spike and waves(29.5%),were more predominant in frontal location when they were awake.During sleep,sharp waves generalized or the amplitude increased.At the last follow-up,88.6%of patients were in complete clinical remission and EEG in 22.7%of cases was normal.Among them 2 patients had stopped taking antiepileptic drugs(AEDs)and 6 patients were reducing the doses of AEDs.EEG Was abnormal only in sleep or decreased synchronization.The patients were more responsive to earbamazepine combined with sodium valproate(P<0.01).Conclusion Special partial epileptic syndrome is age-related,having excellent prognosis,which might origin from the frontal lobe.

ObjectiveTo observe the effect of telmisartan on plasma levels of inflammatory cytokine in coronary artery disease (CAD) patients complicated with diabetes mellitus and hypertension after percutaneous coronary intervention.MethodsFifty CAD patients who just had undertook angioplasty and implanted stents were randomly divided into two groups, the test group (telmisartan group, n=25) and control group (perindopril group, n=25). After treatment, patients were followed-up for 6 months; plasma samples were collected from each patient before and after percutaneous coronary intervention. Then plasma levels of C-reactive protein (CRP), monocyte chemoattractant protein-1 (MCP-1) were measured by enzyme-linked immunoassay. The changes of cholesterol, fasting plasma glucose (FPG), insulin and insulin resistance index (IRI) were observed.ResultsAt the end of 6 months, plasma levels of CRP and MCP-1 of patients in the test group significantly declined (P<0.01), and showing a inversely correlation with FPG (P<0.01), and FPG, insulin and IRI also declined. In the control group, only CRP and MCP-1 declined (P<0.05). Meanwhile, the frequency of cardiovascular events in test group was significantly lower than that in the control group.ConclusionTelmisartan can decline plasma levels of CRP, MCP-1 and frequency of cardiovascular events as well as increasing insulin sensitivity and improving glucose metabolism to unstable angina patients.

Objective To study the clinical effect of celecoxib combined with radiotherapy in the treatment of malignant glioma tumor of brain. Methods A total of 80 patients diagnosed as brain malignant tumor were randomly divided into control group (n=40) and observation group (n=40). The control group received three-dimensional conformal radiotherapy for 5 times a week, with dosage 4-6 Gy each time. Base on control group, the observation group received celecoxib 0.2 g/time twice a day during treatment.After 6 weeks,the clinical efficacy,immune function,quality of life and the serum levels of metalloprotease-9 (MMP-9), transforming growth factor-β (TGF-β) and vascular endothelial growth factor (VEGF) were compared between two groups of patients. Results After 6 weeks of the current therapy, the total effective rate was better in observation group than that of control group(P<0.05).After treatment,the levels of CD3+,CD4+and CD4+/CD8+increased in the both groups.The level of CD8+decreased,and the changes of CD3+,CD4+and CD4+/CD8+were significantly higher after treatment in the observation group than those of control group, but CD8+was lower in the observation group than that of control group (P < 0.05). Meanwhile, the levels of MMP-9, TGF-β and VEGF were significantly decreased after treatment in both two groups.The levels of MMP-9,TGF-β and VEGF were significantly lower after treatment in the observation group(P<0.05).The life quality was significantly better and the overall efficiency of the improvement of the disease was higher in observation group than those of control group(P<0.05).Conclusion Celecoxib can enhance the radiosensitivity of glioma patients.The combined treatment can improve the clinical efficacy and quality of life of patients.Moreover,it improves patients'immune function and reduces the levels of VEGF,TGF-β and MMP-9.

Diabetes is a global threat threatening human health in the world, with an increasing incidence rate in recent years. The disorder of glucose metabolism is one of the major factors. As relevant glucose metabolic enzymes such as alpha-glucosidase, glucose-6-phosphatase (G-6-P), glycogen phosphorylase (GP) and glycogen synthase kinase-3 (GSK-3) get involved in and control the process of glucose metabolism, the regulation of the activity of glucose metabolic enzymes is of significance to the treatment of diabetes. Traditional Chinese medicines (TCMs) have been widely researched because of their low toxicology and high efficiency, and many extracts and components from TCMs have been proven to be regulators of glucose metabolic enzymes. Compared with anti-diabetic western medicines, anti-diabetic TCMs feature safety, reliability and low price. This essay summarizes the anti-diabetic effect of TCMs on regulating glucose metabolic enzymes.
Animals ; Diabetes Mellitus ; drug therapy ; enzymology ; metabolism ; Drugs, Chinese Herbal ; analysis ; therapeutic use ; Enzyme Activators ; analysis ; therapeutic use ; Enzyme Inhibitors ; analysis ; therapeutic use ; Glucose ; metabolism ; Glucose-6-Phosphatase ; metabolism ; Glycogen Synthase Kinase 3 ; metabolism ; Humans ; Hypoglycemic Agents ; analysis ; therapeutic use ; alpha-Glucosidases ; metabolism