Vascular endothelial growth factor (VEGF) plays essential roles in embryonic development and angiogenesis of a variety of diseases.In recent years,the neuroprotective effect of VEGF is increasingly receiving attention.The expression of VEGF is upregulated during hemorrhagic cerebrovascular disease.It plays a protective role in cerebral hemorrhage,but its role is controversial in subarachnoid hemorrhage.The relationship between VEGF and hemorrhagic cerebrovascular disease is both close and complex.More research is needed to clarify the relationship between them and to provide new clinical ideas for the prevention and treatment of this kind of disease.

Objective To approach the therapeutic effect of modified Linggui Zhugan decoction for treatment of patients with type Ⅱ cardiorenal syndrome (CRS).Methods Thirty patients with CRS admitted to Affiliated Hospital of Tianjin Institute of Chinese Medicine were selected. By a random number table and double-blind method, they were divided into two groups: treatment and control groups, 15 cases in each group. The patients in both groups were treated with conventional western medicine, and those in the treatment group were additionally given traditional Chinese medicine (TCM) modified Linggui Zhugan decoction (including the following ingredients: Poria 30 g, Cinnamomi Cortex 10 g, Atractylodis Macrocephalae Rhizoma 15 g, Glycyrrhizae Radix et Rhizoma Preparata cum Melle 10 g, Morindae Officinalis Radix 15 g, Arecae Pericarpium 30 g, Astragali Radix 30 g, Zingiberis Rhizoma 10 g, Descurainiae Semen 15 g), one dose daily for consecutive 30 days. Before and after treatment the changes in levels of B-type natriuretic peptide (BNP), serum creatinine (SCr), blood urea nitrogen (BUN), amount of urine, clinical efficacy and TCM syndrome score efficacy were observed in two groups.Results After treatment, the levels of BNP, SCr, and BUN were significantly decreased, while urine volume was obviously increased compared with those before treatment in the two groups, and the degrees of changes in the treatment group were superior to those in control group [BNP (ng/L): 297.3±75.1 vs. 344.2±56.3, SCr (μmol/L): 139.7±62.1 vs. 154.4±39.7, BUN (mmol/L): 10.1±6.4 vs. 13.2±8.7, urine volume (mL/d): 847.2±32.7 vs. 786.4±13.6, allP < 0.05]. The total effective rates of patients and TCM syndrome scores in treatment group were significantly higher than those in control group [both 86.7% (13/15) vs. 66.7% (10/15), bothP < 0.05]. Conclusions Modified Linggui Zhugan decoction can alleviate the symptoms of yang deficiency of heart and kidney and heart failure due to the attack of heart by retained fluid, and can also ameliorate the complicated renal function impairment; the therapeutic effect of integrative traditional Chinese and western medicine for treatment of patients with type Ⅱ CRS is superior to that of conventional western medicine treatment.

Objective To study the optimum extration process of polysaccharides from Meretrix meretrix Linnaeus.A spectrophotometry method for content determination of polysaccharides from Meretrix meretrix Linnaeus was established with Phenol-sulfuric acid coloration.Methods The samples were extracted by ultrasonic wave,dissolved in 50mL,4mol?L~(-1) HCl and heated to hydrolyze by acid.The contents of polysaccharides from Meretrix meretrix Linnaeus were determined by spectrophotometry at 490nm.Results Extracted for thirty minutes by ultrasonic wave,two times,and acidic hydrolysis were the optimum technology.The calibration curve was linear over the range of 4.848～24.242mg?L~(-1).The regression equation was C=28.69A+0.229(r=0.9992).The average recovery rate was 100.67% and RSD was 2.83%(n=6).Conclusion The method was proved to be simple,accurate and reliable,and could be used to extract and determine the polysaccharides from Meretrix meretrix Linnaeus.

A total of twenty eight HSV isolates from the patients were typed by using HSV typespecific monoclonal antibodies (McAb). Comparison of typing results with McAb in three Sero-lminunological methods with molecular hybridization indicated 100% concordance in the results. But the typing rates were quite different among the various methods (ELISA 64%, IFA and EIA 82%, Hybridization 100%). The results demonstrate that the molecular hybrydization with HSV type-specific probes was highly sensitive and specific, and the method of EIA with HSV type-specific McAb was accurate, cheap, rapid and practical.

BACKGROUND: Umbilical cord blood mesenchymal stem cells (UCB-MSCs) can differentiate into various types of cells under certain condisions, and easily proliferate in vitro. However, UCB-MSCs have long establishment time and low frequency.OBJECTIVE: To in vitro isolate and culture UCB-MSCs, and induce its differentiation into osteoblasts.DESIGN, TIM E AND SETTING: The in vitro cytological study was performed at the Laboratory of the Medical College of Qingdao University from June 2008 to January 2009.MATERIALS: UCB was obtained from term normal delivery women at the Department of Gynaecology and Obstetrics, Qingdao Municipal Hospital.METHODS: Human UCB-MSCs were isolated and cultured in vitro by Percoll density gradient. When reached 90% confluency,UCB-MSCs were digested by trypsin for subculture. At the third passage, UCB-MSCs at 1×106 were incubated. When reached 50% 60% cenfluency, UCB-MSCs were treated with DMEM supplemented with 0.1 μmol/L dexamathasone, 10 mmol/Lβ-sodium glycerophosphate and 50 μmol/L vitamin C. UCB-MSCs in the control group were incubated in low glucose DMEM.MAIN OUTCOME MEASURES: Growth and proliferation of MSCs were observed under the inverted microscope. Cell surface marker expression and cell growth curve were measured by flow cytometry. Cell ultrastructure was observed under the transmission electron microscope. Differentiation of UCB-MSCs into osteoblasts was determined by Won Kossa staining and alkaline phosphatase staining.RESULTS: Primary cultured UCB-MSCs had similar morphology to bone marrow mesenchymal stem cells. After passage, cell morphology was even, presenting spindle shape. UCB-MSCs at passage 3 highly expressed CD29, CD44, CD13, but did not express CD34. Growth latency was 2-3 days. Cells entered logarithm proliferation phase at days 3-4, and platform phase 1 month later. Nuclei presented round or irregular, with clear nuclear membrane, 1-2 nucleoli, rough chromatin, abundant organelles and microvilli. UCB-MSCs at passage 3 were gradually confluent following 3 days of osteogenic induction, with the presence of pavement-stone shape. 14 days later, calcified nodules by Von Kossa staining, and cells were positive for alkaline phosphatase staining. In the control group, no calcified nodules were found, and cells were negative for alkaline phosphatase staining.CONCLUSION: UCB-MSCs can be successfully isolated by Percoil density gradient, and induced to differentiate into osteoblasts in vitro.

Aquaporin4(AQP4),a member of the aquaporin family,is mainly expressed inastrocytes end-feet in the central nervous system. A large number of experimental studies have show n that AQP4 expression plays an important role in the occurrence, development, and regression of brain edema after different types of stroke. In addition, the AQP4 expression can affect the development process of cerebrovascular disease through the mechanisms such as affecting the integrity of the blood-brain barrier and promoting astrocyte migration, nerve regeneration, and neuroinflammatory response. Investigation of regulation mechanisms of AQP4 in transmembrane transport of substance in brain and intracel ular and extracel ular environmental balance and its expression in model of cerebrovascular disease have an important significance for understanding the occurrence, development, protection, and treatment of clinical cerebrovascular disease.

Spontaneous intracerebral hemorrhage is the most deadly type of stroke.Its 30-day mortality rate is nearly 40％.More than 30％ of patients with spontaneous intracerebral hemorrhage will have hematoma growth,which lead to poor outcomes.In addition to the classic predictors,some inaging signs also have important implications for identifying hematoma growth,such as swirl sign,blend sign,and black hole sign on nonenhanced CT scans,extravasation sign on contrast-enhanced CT scans,spot sign on CT angiography,leak sign revealed by the modified imaging method,as well as the spot sign on CT peffusion imaging.

BACKGROUND:Currently, bone marrow mesenchymal stem cel s can differentiate into nerve cel s via many approaches. Different methods for inducing bone marrow mesenchymal stem cel s differentiating into nerve cel s have different ratios. OBJECTIVE:To investigate the difference between chemical method and co-culture method to induce the differentiation of rat bone marrow mesenchymal stem cel s into nerve cel s. METHODS:Rat bone marrow mesenchymal stem cel s were isolated and purified using whole bone marrow culture method, and then randomly divided into two groups:chemical group,β-mercaptoethanol was added;co-culture group, co-cultured in a Transwel chamber. RESULTS AND CONCLUSION:Visible protrusions from induced cel s showed radiation growth at 1 week of induced culture, and neuron-specific enolase staining was positive at 2 weeks of culture. Star-like structure of nerve cel s was visible in the co-culture group within 4-5 days of culture, and then more protrusions formed. Meanwhile, the positive rate of neuron-specific enolase was (70.82±2.46)%. After 6-7 days of culture, neuron-like cel s formed and were interconnected in the chemical group;while, the positive rate of neuron-specific enolase was (52.37±1.83)%. These findings suggest that cel microenvironment plays a leading role in the differentiation of bone marrow mesenchymal stem cel s into nerve cel s, and chemical induction method is inferior to the co-culture method.

Objective To investigate the change of leptin,nitric oxide (NO) and interleukin-6 (IL- 6) levels in serum of children with Kawasaki disease (KD) and the possible relationship between leptin,NO and IL-6 levels,explore the role of leptin,NO and IL-6 in the pathogenesis of KD.Methods Fourty-five children with KD were studied.Twelve of them had coronary artery lesions and 33 had non-coronary artery lesions;thirty healthy children and 18 children with juvenile idiopathic arthritis or Henoch-Scholeion purpuru were enrolled as control subjects.Serum was collected from each patients during acute stage of KD and remission.Leptin,NO and IL-6 contents were detected by radioimmuno-assay and spectrophotometry and enzyme-linked immunoserbent assay.Meanwhile,C-reactive protein (CRP) were examined.Results ① The concentrations of serum leptin,NO,IL-6 and CRP in children with KD were significantly higher in the acute stage of KD than those at clinical remission and those of the normal control group (q=26.24,25.23; 21.38,31.30;35.37,33.68;16.32,15.66;P＜0.01,respectively).No significant differences in serum leptin, IL-6 and CRP were found between the clinical remission group and the normal control group (q=1.02,1.04, 0.61,P＞0.05,respectively);The concentrations of serum NO were significantly higher at clinical remission group than those of the normal control group (q=11.31,P＜0.01).② There was no significant difference in the concentrations of serum leptin,IL-6 and CRP at the acute stage of KD than those in patients with and without coronary artery lesions (q=1.17,1.92,1.60,P＞0.05).The concentrations of serum NO were significantly higher at the acute stage of KD with coronary artery lesions than those of KD without coronary artery lesions (q=6.91,P＜0.01).③ The concentrations of serum leptin in children with juvenile idiopathic arthritis or Henoch-Scholeion purpura were signifietantly higher than those of the normal control group (t=13.26,P＜ 0.01).No significant differences in serum leptin were found between children with juvenile idiopathic arthritis or Henoch-Seholeion purpura and children with KD (t=1.28,P＞0.05).④ Correlation was found between serum leptin values and levels of the following parameters (P＜0.01);NO (r=0.69),IL-6 (r=0.55),CRP (r=0.42).However,there were no associations between leptin and leukocytes (r=0.21,P＞0.05) or serum albumin level (r=-0.24,P＞0.05).Association was found between serum NO and IL-6 (r=0.45,P＜0.01)or CRP(r=0.49,P＜0.01).Conclusion These results suggest that leptin,NO and IL-6 may have a role in the immunoinflammatory process of KD,especially in the acute phase.Further in vivo and in vitro studies are needed to establish the roles of leptin,NO and IL-6 in the pathogenesis of KD.

Objective To construct the expressing vector of shRNA/mrp1 and study its expression in vitro. Methods 64bp oligonucleotides of pSUPER and the targeted senquence of siRNA/mrp1 were synthesized and annealed to form duplex strand, then were cloned into pSUPER to construct pSUPER-shRNA/mrp1 vector. Competenced Ecoli was transfected by vector of pSUPER-shRNA/ mrp1 to screen the positive clones for sequencing and extracting plasmid. The plasmids extracted were used to transfected HepG2/mrp1 cells with a control groups by negative vectors. The expression of mrp1 mRNA and MRP1 was measured by real-time PCR and resistance of HepG2/mrp1 by flowcytometry. Results pSUPER-shRNA/mrp1 was established successfully and was sequenced to test its accuracy. Expression of mrp1 mRNA in HepG2/mrp1-si was lower than that in HepG2/mrp1 (1-fold vs 179.76-fold, P<0.001). Compared to HepG2/mrp1, the expression of MRP1 in HepG2/mrp1-si was lower (11.2% vs 97.6%, P<0. 05). The sensitivity of HepG2/mrp1-si to adiramycin was higher than that of HepG2/mrp1(45.0-fold vs 1.2-fold, P<0.01). Meanwhile, the accumulation of DNR in HepG2/mrp1-si increased significantly as compared with the control (78.58 % vs 38.44%,P<0.05).Conclusion Vector of pSUPER-shRNA/mrp1 can be constructed by the technique of enzymatic incision. The multidrug resistance of HepG2/mrp1 can be reversed by RNA interference.