Aim To study the protective effects of Ganoderma lucidum polysaccharides peptide(GLPP)on the human umbilical vein endothelial cells(HUVECs)injured by reactive oxygen species(ROS),derived from tert-butylhydroperoxide(t-BOOH).Methods Vascular endothelial cells were obtained from infant umbilical cord for primary cultivation,identified by endothelial cell adhesion molecule-1(CD31)by fluorescence microscope.HUVECs were injured by ROS,derived from t-BOOH.The survival rate of cells was measured by Cell Counting Kit-8 assay.Hoechst 33258 nucleus staining was used to observe the early apoptosis injury by ROS.And the activation of Caspase-3 was detected by spectrophotometry.Electron microscopy was used to show the morphological changes in HUVECs.Results GLPP(6.125,12.5,25,50,100 mg?L-1)could inhibit the apoptosis of HUVECs by ROS.The survival rate of HUVECs was increased by GLPP and the percentage of cell apoptosis was decreased by it too.GLPP decreased the activation of Caspase-3 of HUVECs up-regulated by ROS.Electron microscope showed that the organelle such as mitochondria injury by ROS could be relieved by GLPP.Conclusion GLPP can prevent human umbilical vein endothelial cells from oxidative injury.

Objective To analyze the pregnant outcome of women with chronic hypertension,and to investigate the high risk factors associated with harmful maternal and perinatal prognosis.Methods Of the 14127 deliveries in Peking University First Hospital from Jan 2001 to Dec 2005,121 pregnant women with chronic hypertension were identified and divided into two groups:chronic hypertension with and without preeclampsia(group PE,64 cases;group N-PE,57 cases).The maternal and perinatal outcomes of the two groups and the high risk factors for adverse maternal and perinatal prognosis were analyzed.Results(1) The incidence of pregnancy with chronic hypertension was 0.86%(121/14127).(2)The incidences of placental abruption,pulmonary edema and retinopathy in groups PE and N-PE were 16%(10/64)vs 2% (1/57),11%(7/64)vs 0 and 41%(26/64)vs 16%(9/57,P

BACKGROUND: To search for an alloxenogeneic bone with good load bearing function and osteoblastic activity for treating bone defects is an important study issue. We have made a comparative study on its biome chanical characteristics and found that there was no significant difference in maximum load stress, maximum pressure as compared with fresh bone of the same size. Clinicians are concerned about the osteoblastic activity and whether the osteoblastic activity can be reserved after human allogenous a cellular bone (HAB) loaded with bone marrow stromal cells (BMSCs). OBJECTIVE: To investigate the experimental effect of HAB loaded with induced BMSCs, and observe the cellular adherence and growth as well as detect its osteoblastic activity. DESIGN: Single sample experiment. SETTING: Second Affiliated Hospital of Harbin Medical University. MATERIALS: This experiment was conducted at the Experimental Center of the Second Affiliated Hospital of Harbin Medical University between January 2003 and August 2004. HAB was obtained from fresh corpse iliac bones (donated voluntarily). METHODS: Connective tissues and cell compounds of the iliac bones were removed by processing with hydroperoxide andether solution and sterilized for preparing HAB. BMSCs from living femoral shaft bone marrow were cultured immediately in ordinary and mineralized medium containing DMEM, fetal bovine serum, dexomethasone, β-glycerophophate and ascor bic acid. Proliferation and differentiation of bone stromal cells were deter mined by detecting the level of alkaline phosphatase (ALP) and osteocalcin (OCN) in the culture medium. Induced bone stromal cells solution was condensed and implanted within HAB scaffold. Cellular osteoblastic activ ity was determined through morphological observation under the light mi croscope and electron microscope as well as biochemical index detection. MAIN OUTCOME MEASURES: ① Detection results of ALP and OCN of BMSCs/HAB composite. ② Histological observation results of BMSCs/ HAB composite. RESULTS: ① Iliac bone block cells were cleaned with good reservation of bone matrix. ② The level of ALP and OCN of MSCs was higher after in ducing for 8 days than that in control group [MSCs after induction: (181.54±40.01) nkat/L, (7.2±1.3) μg/L. There was no method to detect the level in control group, P ＜ 0.05]. ③ BMSCs were adhered and grew well in HAB scaffold. CONCLUSION: HAB loaded with induced BMSCs has an excellent os teogenic function in vitro and shows an effective potential as a good bone tissue engineering material.

Objective To evaluate the role of fibroblast growth faetor-2 (FGF-2) in neuropathic pain (NP) in rats.Methods One hundred male Sprague-Dawley rats,aged 7 weeks,weighing 200-250 g,were randomly divided into 4 groups (n=25 each): sham operation group (group S),group NP,phosphate buffer solution (PBS) group and FGF-2 antibody group (group Ab).FGF-2 antibody 18 μg (40 μl) was injected intrathecally at 1,6,9,13,16 and 20 days after operation in group Ab,while the equal volume of PBS was injected intrathecally in group PBS.Spinal nerve injury was produced by exposing the sciatic nerve and its branches and ligation and transection of tibial nerve and common fibular nerve.Pain behavior was assessed at 1 day before operation and at 1,4,7,14 and 21 days after operation.The paw withdrawal mechanical threshold (PWMT) was measured.The animals were then sacrificed and the lumbar segment (L4-6) of the spinal cord was obtained for determination of the contents of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the spinal cord.Results Compared with group S,the PWMT was significantly decreased and the contents of TNF-α and IL-6 in the spinal cord was significantly increased in groups NP,PBS and Ab (P ＜ 0.05).The PWMT was significantly higher and the contents of TNF-α and IL-6 in the spinal cord were significantly lower in group Ab than in groups NP and PBS (P ＜ 0.05).Conclusion FGF-2 is involved in the occurrence and development of NP and induction of the inflammatory response in the rat spinal cord in involved in the mechanism.

Objective To investigate the effects of clinical pathway-based teaching methods in clinical teaching of hematology. Methods Interns, which studied in department of hematology, were classified non-randomized into two groups. The interns in experimental group received idiopathic thrombocytopenic purpura (ITP) clinical pathway-based teaching. And the interns in the control group received the seventh edition textbook of medicine-based traditional teaching. All interns were oral tested when they finished two weeks period clinical studied of hematology. Meanwhile, a questionnaire designed for clinical pathway-based teaching was included in this test only for the experimental group. Results There were 32 interns recruited in the group of clinical pathway-based teaching and 38 interns in the control group of traditional teaching. There were statistically significant differences between the two groups in the fields of the key points about history of diseases ( = 0.0017), assessment of diagnostic criteria and differential diagnosis ( =0.0074), selection of laboratory examination items ( <0.0001) for ITP. The group of clinical pathway-based teaching achieved higher scores than the control group. However, there was no statistically significant difference between the two groups in the field of selection of first-line treatment measure of ITP ( = 0.1155) . Moreover, the control group achieved higher scores than the group of clinical pathway-based teaching in the field of how to treat ITP patients with first-line treatment failure ( =0.0003) . In addition,there were 93.8%interns in the experimental group accepted the new clinical pathway-based teaching tool in the survey. The open-ended question survey showed the clinical pathway teaching method was more simple, intuitive and standardizing than the traditional one. Conclusion The ITP clinical pathway, as an interns teaching tool of hematology, is helpful for understanding more clearly the diagnosis and treatment of ITP. However, an in-depth explanation is necessary combination with textbook of medicine study in the clinical teaching of ITP.

Objective We investigated female patients with idiopathic inflammatory myopathy (ⅡM),including polymyositis (PM) and dermatomyositis (DM) to understand if the activity and treatment of ⅡM affected the fetus outcomes as well as the correlation between pregnancy and the pathogenesis of ⅡM.Methods Questionnaires were designed,including patients' fertility,the activity and treatment of ⅡM during pregnancy and after childbirth,and the fetal outcomes.The questionnaires were conducted on 75 female patients (22-53 years old),who once hospitalized at Peking Union Medical College Hospital from 2007 to 2013.The mutual effect between the ⅡM disease activity,pregnancy and the fetus were compared by Chi square.Results Seventy-five patients (19 PM,56 DM) had 144 pregnancies,but only 18 of them became pregnant after the onset of the disease.There were two patients had onset of DM during pregnancy,and two patients suffered from DM after delivery or abortion within one month.The rates of abnormal pregnancy in patients with pregnancy before the onset of ⅡM was significantly lower than those with pregnancy after the onset of ⅡM (x2=10.21,P=0.001).Of the 18 pregnancies after the onset of ⅡM,7 had active ⅡM during pregnancy,3 of them flareddue to pregnancy.The 5 abnormal pregnancies (5/7) were seen in seven pregnancies totally,including two spontaneous abortion,one embryo diapause and two premature births.While of the other 11 pregnanciesin stable condition had only 2 abnormal pregnancies (2/11),including one fetus ended prematurely and one embryo diapause.The fetal outcomes were significantly related to the status of illness (x2=5.103,P=0.024).Two patients became pregant after the onset of the disease,both of them had stable disease before pregnancy.However,the disease became active during the pregnancies.One of them flared with rash during two pregnancies.The fetal end in embryo diapause and premature birth (33+4 weeks),respectively.Another flared with muscle weakness at five months of gestation.The fetal ended in premature birth (36+3 weeks).Two patients had onset of DM within one month following the delivery and voluntary abortion.Patients with active disease had taken medium to large doses of prednisone (30-60 mg) orally,in combination with hydroxychloroquine (0.2 bid) or without.In addition,two patients administrated with intravenous gamma globulin (IVIG) at low dose,whose illness condition had been well controlled.Conclusion Pregnancies after the onset of ⅡM tend to end with spontaneous abortion,embryo diapause,premature birth.The fetal outcomes are significantly related to disease activity.When ⅡM flared during pregnancy,IVIG might be the most effective and safe treatment,combined with corticosteroids.

BACKGROUND: Previous studies have verified that under normal culture of neural stem cells (NSCs), folic acid can accelerate proliferation of NSCs by phosphorylation of mitogen activated protein kinase path activation ERK1/2. OBJECTIVE: To investigate the effect of folic acid on NSC extracellular signal regulatory protein kinase pERK1/2 under hypoxic condition. METHODS: NSCs from Neonatal rats were cultured in vitro by serum-free culture method, and incubated in a flask at 1×10~8/L. Except normal control group, self-made hypoxia equipment was used in the hypoxia model, folic acid deficiency and folic acid supplemented groups at day 3. At 37 ℃, hypoxia culture was conducted in the thermostat for 6 hours. The contents of folic acid were 4 mg/L, 4 mg/L, 0.65 mg/L, 8 mg/L in the four groups. Cells following 6 days were collected to count the density using trypan blue. RT-PCR was utilized to detect pERK1/2 mRNA expression. Western blot assay was employed to determine pERK1/2 protein expression. RESULTS AND CONCLUSION: Compared with the normal control group, the proliferation of NSCs and the expression of ERK1/2mRNA and pERK1/2 protein were decreased significantly in the hypoxia model group. Compared with the hypoxia model group, the proliferation of NSCs and the expression of ERK1/2mRNA and pERK1/2 protein were increased in the folic acid supplemented group, whereas decreased in the folic acid deficiency group. There were significant differences among groups (P < 0.001). Above-described results verified that folic acid supplementation can activate ERK1/2 phosphorylatin and accelerate proliferation of NSCs under hypoxia condition.

Objective To study microbial flora on skin lesions and their antim icrobial susceptibilities and to conduct a topical treatment trial in atopic der matitis (AD). Methods Seventy one patients with AD were diagnosed according to the criteria of Hanifin-Rajka′s in departments of dermatology in China-Japan Friendship Hospital and Beijing Children Hospital during November,2001 to March, 2002. Bacteria and fungi were examined by culture and microscopy. Among 71 case s, 66 were divided into two groups randomly with one group receiving 1%econazol e+0.1%triamcinolone acetonide cream topically twice daily for 4 weeks, and the control group receiving 0.1%hydrocortisone butyrate ointment. Severity of AD w as measured by SCORAD index. Results It was shown that AD mainly affected infant s and children with a prevalence of 73.24%in patients less than 12 years old. T he rates of bacterial and fungal colonization were 53.51%, 1.41%, respectively on skin lesions. S.aureus was detected in 35.21%of patients with AD. Antimicro bial susceptibility test indicated that S.aureus was sensitive to rifampin, vanc omycin, amikacin, ciprofloxacin, cefazolin, cefuroxime, etc. It was found that 1 %econazole+0.1%triamcinolone acetonide cream had significantly more advantage over hydrocortisone butyrate ointment in the therapeutical effects (P

OBJECTIVE:To prepare paeonol-HP-β-cyclodextrin (PAE-HP-β-CD) inclusion compound and to optimize its pre-scription technology. METHODS:PAE-HP-β-CD was prepared by freeze drying method and validated. Using inclusion rate as in-dex,main drug-accessory ratio,inclusion time,inclusion temperature and stirring speed as factors,the preparation technology was optimized by central composite design-response surface methodology. RESULTS:Prepared PAE-HP-β-CD underwent phase transfor-mation. The optimal inclusion technology was as follows as main drug-accessory ratio of 3.39∶1,inclusion temperature of 50 ℃, inclusion time of 3.2 h, stirring speed of 350 r/min. Relative error between measured value (87.46%) and predicted value (89.12%) of inclusion rate was 1.86%(n=6). CONCLUSIONS:PAE-HP-β-CD inclusion compound is prepared successfully, and its prescription technology is stable and feasible.

Objective:To prepare nanobody-based immunotoxin BI7D12-PE38KDEL targeting EGFR and to examine its cytotoxicity against EGFR positive tumor cells.Methods:By using molecular cloning strategy,prokaryotic expression construct of pET28a-BI7D12-PE38KDEL was generated which consisted of nanobody 7D12 targeting EGFR in the form of a divalent fused with PE38KDEL,a truncated form of pseudomonas exotoxin A via a flexible peptide(G4S)4,and then transformed into E.coli BL21(DE3).Protein expression was induced by adding IPTG,purified by Ni-affinity column chromatography,and verified by Western blot.The binding capacity of the resulted immunotoxin to EGFR-positive cells A549,HT29,MCF-7 and EGFR-negative cells CEM,Jurkat were determined by flow cytometry assay,and its cytotoxicity against the target cells was examined.Briefly,tumor cells were treated with different dosage of the immunotoxin,and the killing efficacy of BI7D12-PE38KDEL on these cells were assessed by WST-1 assay after 72 hours.Results:The SDS-PAGE and Western blot results showed the recombinant immunotoxin BI7D12-PE38KDEL was successfully prepared,and majority of them was expressed in soluble form.BI7D12-PE38KDEL could selectively bind to EGFR-positive cells of A549,HT29,and MCF-7.More importantly,the immunotoxin exhibited much more significant killing effect on these EGFR positive cells compared to the negative control group of CEM and Jurkat cells(P<0.01).Conclusion:In the current study,the nanobody-based immunotoxin BI7D12-PE38KDEL targeting EGFR was successfully prepared and exhibited a superior inhibition effect for the growth of EGFR-positive cells.