BACKGROUND: The observation of vascular endothelial growth factor gene expression of cerebrovascular diseases and ultrastructure of cells may be helpful to understand angiogenesis and its relative cellular factors involved in the pathogenesis at cellular and molecular levels. OBJECTIVE: To investigate the method of culture of human cerebral cap illary endothelial cell by separation of capillary fragment in vitro, and to ob serve vascular endothelial growth factor gene expression and ultrastructure of cells. DESIGN: A randomized controlled research on technique and method. SETTING: The neurosurgery department of a general hospital of a military area command of Chinese PLA and the neurosurgery department of a college hospital. PARTICIPANTS: Eighteen patients with arteriovenous malformation of brain(Spetzler Ⅱ-Ⅲ grade), as confirmed by aortocranial angiography before operation, in the Department of Neurosurgery, General Hospital of Shenyang Military Area Command of Chinese PLA were included. The material was obtained from fresh integrated specimen of arteriovenous malfor mation of brain with surrounding fresh brain tissues during the opera tion. Capillary endothelial cell was separated by homogenate, filtration and enzymatic digestion techniques. Cells grew well in culture flask and were divided into 4 groups(hypoxia state for 2, 4, 8 hours groups and control group). Each group contained four flasks.METHODS: Simulation of anoxia condition: volume faction 0.95 N2 and volume fraction 0.05 CO2. Expression of factor Ⅷ related antigen in cells was detected by immunohistochemistry. mRNA expression of vascular endothelial growth factor on endothelial in every group was observed by RT-PCR, protein content of vascular endothelial growth factor in supernatant detected by enzyme-linked immunoadsordent assay, and cellular ultrastructural change observed by transmission electron microscopy.MAIN OUTCOME MEASURES: mRNA expression of vascular endothelial growth factor on endothelial cell and protein content of vascular endothelial growth factor in supernatant in control group and every hypoxia groups; cellular ultrastructural changes.RESULTS: Under phase contrast microscope, cultured living cells had mono-layer pebble-like typical character. More than 90% of were factor Ⅷrelated antigen(FⅧ-RA) staining positive. mRNA and protein expression of vascular endothelial growth factor in hypoxia 4 hours group was 0.98 ±0. 19,( 180. 77 ± 20. 15) ng/L, which was significantly higher than in control group [0, (26. 20 ± 6.33) ng/L, P ＜ 0.01 ]. Eight hours later, expression decreased [(0. 35 ±0.07), (31.68 ±8.34) ng/L]; swollen mitochondrion, dilated endoplasmic reticulum, and lysosome vesiculation were found.CONCLUSION: Humane cerebral capillary endothelial cell can be cultured by separation of capillary fragment, which is easy to operate and the cellular purity is reliable. In the early stage of ischemia and hypoxia, expression of vascular endothelial growth factor is not enough to maintain cellular ultrastructure integrity. Cells may be injured along with the prolong of hypoxia.Zhao MG, Tang T, Gao YZ, Pu PY, Wei XZ. Culture of human cerebral capillary endothelial by separation of capillary fragment and the observation of vascular endothelial growth factor gene expression and cell ultrastructure. Zhongguo Linchuang Kangfu 2005; 9(21):211-3 (China) [www. zglckf. com]

This article investigates the effect of regulating wei qi and tonifying brain on insomnia from two aspects. It provides theoretical basis for clinical application of regulating wei qi and tonifying brain. First of all, this method regulates the sleep function to keep the body’s yin-yang balance by regulating wei qi. Wei qi is the defense qi in the body and has close relationship with sleep. Wei qi can regulate sleep by the excitement and depression of Yin and Yang Heel Meridians. Then, this method also can improve the brain function for insomnia treatment by dredging Governor Channel and tonifyjng marrow, tonifyjng brain and calming the nerves. On basis of inheriting traditional theory, this article expounds the theory of treating insomnia by regulating wei qi and tonifying brain and verifi es its stable therapeutic effect and safty by clinic study.

0.05),and the activities of SOD in those groups were statistically lower(P

Objective To explore the correlation of S100 protein with the prognosis of patients with primary retroperitoneal liposarcoma. Methods Analyzed the clinical data about 108 patients with primary retroperitoneal liposarcoma managed with surgery from January 2009 to June 2014. All patients were followed up. Patients were divided into S100-positive group(58 patients) and S100-negative group (50 patients) according to the immunohistochemical staining results. The overall survival time and all clinical data between two group were compared. Results All patients with primary retroperitoneal liposarcoma received radical surgical resection for the first time. The overall 5-year recurrence rate were 88.9%(96/108), and the median recurrence time was 32.7 months. The 1-year, 2-year, and 5-year recurrence rates of the S100- positive group were 25.9% (15/58), 53.4% (31/58), 96.6% (56/58), respectively, and the median recurrence time were 26.2 mouths. The 1-year, 2-year, and 5-year recurrence rates of the S100-negative group were 10.0%(5/50), 36%(18/50), 80.0%(40/50) and the median recurrence time were 40.0 mouths. Log-rank test showed that S100 protein expression was significantly associated with postoperative recurrence rates (c2=9.931, P=0.002) and survival time (c2=4.571, P = 0.033). The difference between gender, age, removal of the joint organs and tumor size showed no statistical significance on disease special survival (P>0.05). Cox regression analysis showed that S100 protein expression (OR=1.582, 95%CI:1.005-2.491) and histologic subtype (OR=1.531, 95%CI: 1.254-1.870) were independent risk factors of the prognosis of primary retroperitoneal liposarcoma patients. Conclusions S100 protein played a critical role in retroperitoneal liposarcoma carcinogenesis and its expression may be used as a potential survival predictor in patients with primary retroperitoneal liposarcoma.

Objective To observe the clinical efficacy of micro plasma in the treatment of acne atrophic scar on face.Methods 82 cases with facial acne atrophic scar were randomly divided into the observation group and the control group.Each group had 41 cases.The observation group received micro plasma treatment while the control group were treated with ultrapulsed fractional CO2.The treatment was carried out 6 weeks interval, in total 3 times.After each treatment, the curative effect and the side effects were compared between the two groups.Results The total effective rates had no statistical difference between two groups, and the more treatment times, the better effect.The observation group had less adverse effects, which the duration of main side effects was shorter in the observation group than in the control group, with statistically significant difference.Conclusion The micro plasma technology on treating facial acne atrophic scar was an effective skill with less adverse reactions and worth of wide use.

Objective To investigate the gene expression of vascular endothelial growth factor(VEGF) and ultrastructural changes in cultured endothelial cells from human cerebral microvessels under hypoxic conditions.Methods Human cerebral microvessels were isolated from freshly obtained specimens of normal brain adherent to resected cerebral arteriovenous malformations.The expression of factor Ⅷ-relative antigen(FⅧ-RA) in cultured cells was observed with immunocytochemistry.The level of VEGFmRNA in cells and released VEGF protein in cell supernatant were determined by RT-PCR analysis and ELISA respectively when they were exposed to hypoxic conditions(95% N_2,5% CO_2;two hours,four hours,eight hours) or maintained in basal condition.Ultrastructural changes in cells were also observed by electron microscopy.Results In inverted microscope the cultured cells showed contact inhibition and a rounded cobblestone appearance.More than 90% of them were stained strongly with antibodies against FⅧ-RA.Significant VEGF mRNA and protein accumulated when these cells were exposed to hypoxia for 4 hours.However,their VEGF expression was down-regulated after hypoxia for 8 hours and a number of vesicles and swollen mitochondria were present in the cytoplasm.Conclusion The level of VEGF expression may havesignificant relationship with ultrastructural changes in human cerebral endothelial cells under hypoxic conditions.

Objective To explore the correlation between hyperuricaemia and blood pressure, and blood lipid and glucose. Methods By using simple cluster sampling, 2 branch units from PetroChina Changqing Oilfield Company were selected, and all the 720 subjects with hyperuricaemia (HUA) were assigned to the HUA group; another 620 participants with normal uric acid (UA) level into the normal group. The correlation between blood uric acid and blood pressure,and blood lipid and glucose was assessed by Logistic regression. Results The odds ratio (OR) of those who had 1,2 or 3 abnormal status of hypertension,hyperlipidemia and impaired fasting glucose in the HUA group were much higher than the normal group (OR values were 4. 036,2. 562, and 4. 174, respectively). Logistic regression showed that male, systolic blood pressure ( SBP), GLU, total cholesterol ( TC), triglyceride ( TG), low-density lipoprotein cholesterol (LDL-C) were risk factors of H UA ( OR values were 7. 736,2. 309,1.721,2. 761 and 1. 411,respectively) ,while high-density lipoprotein cholesterol (HDL-C, OR = 0. 211 ) was a protective factor of HUA. Conclusions Gender,blood pressure and blood lipid may have correlation with blood UA. Multiple risk factors should be considered to improve the effectiveness of health education and health promotion.

Objective To analysis the incidence and management of monozygotic twin (MZT) conceived by assisted reproductive techniques (ART). Methods A retrospective analysis of clinical pregnancies and MZT that resulted from ART was performed in Reproductive Medical Center,the First Affiliated Hospital,Wenzhou Medical University between January 2011 and January 2014. Results A total of 5 908 pregnancies were diagnosed: 2 012 twins, 157 high-order multiple pregnancy (HOMP), including 4 quadruplets. Overall, 51 MZT pregnancies were identified of them including 32 cases HOMP and 19 cases MZT. The incidence of MZT resulting from cleavage-stage embryo transfer was similar to blastocyst transfer (P=0.960). The percent of MZT resulting from in vitro fertilization [0.93% (28/3 022)], frozen-thawed embryo transfer [0.87% (13/1 502)] and intracytoplamic sperm injection [0.72% (10/1 384)] did not show statistical significance (P=0.794). The expectantly managed MZT was associated with a significantly greater likelihood of miscarriage [6/19 vs 5.11% (101/1 976)], and low birth weight infant [73.91% (17/23) vs 42.89% (1 453/3 388), P<0.01], when compared with dizygotic twin (DZT) did not undergo selective embryo reduction(SER). In monozygotic (MZ)-triplets with SER to 2 fetuses or to 1 fetus, there was no cases of preterm birth or low birth weight infant observed in MZ-triplets with SER to 1 fetus;when compared with MZ-triplets with SER to 2 fetuses, the low birth weight infant [56.00% (14/25), P=0.021] has statistical significance. The likelihood of the survival of two babies was lower in MZ-triplets with SER to 2 fetuses when compared with non-MZ triplets with SER to 2 fetuses [42.86%(9/21)vs 75.21%(91/121), P=0.003]. Conclusions The incidence of MZT pregnancies following ART is high. It plays a significant role in the occurrence of HOMP. MZT pregnancies are at an increased risk of adverse outcomes , it should transform to a single embryo thansfer (SET) program to reduce them incideuce. Reduction of MZT contained in multiple pregnancies appears to be a safe option.

Objective To compare the laboratory and clinical results between unstimulated in vitro maturation (IVM) and IVM converted from in vitro fertilization (IVF) in polycystic ovarian syndrome (PCOS) and non-PCOS patients.Methods We divided 591 IVM cycles in the First Affiliated Hospital of Wenzhou Medical Univesity from Jan.2008 to Dec.2013 into 4 groups:group A1B1,PCOS patients underwent unstimulated IVM protocol,240 cycles; group A1B2,PCOS patients underwent IVM converted from conventional stimulated IVF protocol,153 cycles; group A2B1,non-PCOS patients underwent unstimutlated IVM protocol,103 cycles; group A2B2,non-PCOS patient underwent IVM converted from conventional stimulated IVF protocol,95 cycles.Multiple linear regression method and binary logistic regression method were used to assess the influence of PCOS and protocols for IVM on laboratory and clinical outcomes.Results The mean number of oocytes retrieved was positively related with PCOS [partial regression coefficient (B)=3.37,P＜0.01].The maturation rate of oocytes was positively related with hCG-prime prior to oocyte aspiration (B=0.05,P=0.010).High-quality embryo rate was positively related with PCOS and IVM converted from IVF (B=0.08,P=0.010; B=0.09,P=0.001),as well as implantation rate related with them (B=0.07,P=0.010; B=0.10,P＜0.01).PCOS and IVM converted from IVF improved hCG positive (hCG＞10 U/L) rate (OR=1.636,95％CI:1.113-2.204,P＜0.05; OR=1.861,95％CI:1.307-2.649,P＜0.05) and the clinical pregnancy rate (OR=1.507,95％CI:1.041-2.240,P＜0.05; OR=1.881,95％CI:1.312-2.696,P＜0.05).IVM converted from IVF protocol decreased the spontaneous abortion rate (OR=0.490,95％CI:0.245-0.978,P＜0.05).Multiple gestation rate and ectopic pregnancy rate were not affected by PCOS condition and protocol used (P＞0.05).Conclusions PCOS and IVM converted from IVF protocol improved the high-quality embryo rate,implantation rate,hCG positive rate and clinical pregnancy rate.IVM converted from IVF protocol reduced the spontaneous abortion rate.PCOS patients may be more suitable for the IVM treatment.No matter PCOS or non-PCOS patients,IVM converted from IVF protocol had better pregnancy outcome than that of unstimulated cycle.

Objective To investigate clinical effect of in vitro maturation(IVM)of immature oocytes transferred from conventional in vitro fertilization embryo transfer(IVF-ET) cycles.Methods From January 2008 to June 2009.medical documents of 155 infertile patients underwent IVF-ET in the Reproductive Medical Center of Fimt Affiliated Hospital of Wenzhou Medical College were analyzed retrospectively.If more than 20 oocytes were monitored after 5-7 days of ovulation induction or follicular developmental retardation were confirmed after 8-13 days of ovulation induction.according to patients' wish,IVM were transferred in 60 cycles(group A).In the mean time.IVF was continued in 95 cycles (group B).The mean dosage of gonadotropin,the cancel lation rate of cycles,the mean numbers of oocytes retrieved and maturation,the rate of fertilization and excellent quality embryos.pregnancy outcome and the incidence rate of ovarian hyperstimulation syndrome(OHSS)were compared and analyzed.Results The rates of embryo transfer were 92%(55/60)in group A and 63%(60/95)in group B,which showed significant differences (P<0.05).In group A,the mean dosage of the gonadotropin,the mean number of oocytes retrieved,the cleavage rate and OHSS rate were(1030±468)U,10±6,82.2%(231/281)and O,and were(1544±338)U,14±4,94.0%(502/534)and 35%(21/60)in group B,respectively,all data abeve exhibited statistieal difference between two groups(P<0.05).However,the rates of fertilization and excellent quality embryos had no significant differences between two groups(P>0.05).In group A,the rate of clinical pregnancy per transfer was 53%(29/55)and multiple pregnancy was 14%(4/29),and were 47%(28/60) and 32%(9/28)in group B,they all had no significant differences (P>0.05).Conclusion IVM of immature oocytes used in conventional IVF cycles not only obtained a high clinical pregnancy rate,but also reduced gonadotropin using dosage and avoided OHSS completely.