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 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.

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.

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.

Adult ; Choristoma ; Humans ; Male ; Mediastinal Cyst ; Neck ; pathology ; Thymus Gland ; pathology

OBJECTIVETo study the therapeutic efficacy of manipulative reduction and percutaneous pin fixation for the treatment of severely displaced distal tibial fractures in children.

METHODSFrom May 2009 to December 2012,56 children with severely displaced distal tibial fractures were analyzed, who had been treated with manipulative reduction and percutaneous pin fixation. Preoperative fractures were confirmed as severely displaced fractures by X-ray apparatus. There were 33 boys and 23 girls,ranging in age from 3 to 14 years, with an average of 10.1 years. All the fractures were closed without vascular or nerve injuries. According to the bone fracture type, under C-arm fluoroscopy, functional reduction was achieved by manipulative reduction. The fractures were fixed with percutaneous pins. Postoperative X-ray confirmed the functional reduction. Follow-up indexes were recorded: intra-operative and postoperative complications,postoperative radiographic examination, lower extremity length and range of ankle motion. Ankle score system of Teeny was used to evaluate ankle function.

RESULTSAll the patients were followed up, and the duration ranged from 3 to 46 months, with an average duration of 19.4 months. According to the Teeny score standard, 35 patients got an excellent result, 7 good and 3 fair. Pin track reaction was found in 4 cases. The X-ray showed all the fractures healed without shortening deformity or epiphyseal arrest at early stage. All the patients could participate in the normal physical activities, having a normal range of motion and excellent strength of the ankle joint.

CONCLUSIONThe method of manipulative reduction and percutaneous pin fixation is a safe and convenient treatment for severely displaced distal tibial fractures in children. It has several advantages as follow: micro-trauma, tiny tissue damage, firm fixation, and the patients can exercise the function of ankle early, suggesting that it is an effective treatment method.

Adolescent ; Adult ; Bone Nails ; Child ; Child, Preschool ; Combined Modality Therapy ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Tibial Fractures ; surgery

Taxa-4(5),11(12)-diene is the precursor for paclitaxel biosynthesis. The diterpenoid paclitaxel (marketed as Taxol), a plant secondary metabolite isolated from yew, is an effective drug widely used in the treatment of numerous cancers. However, further application of taxol has been restricted due to its low yield in plants and the difficulties in extraction. To increase the intact isoprene flux, we constructed the fusion gene plasmid pBgGGTS and individual cassette plasmid pBgGGgTS to enhance the expression levels of geranylgeranyl diphosphate synthase gene (ggpps) and a taxadiene synthase gene (ts) in Coprinopsis cinerea. These two plasmids were separately transformed into C. cinerea LT2 strain, resulting in several putative transformants. Putative transformants were determined by PCR technique, indicating that 5 out of 13 putative transformants transformed by pBgGGTS and 6 out of 13 putative transformants transformed by pBgGGgTS, respectively. Additionally, the Southern blotting analysis of these 10 transformants confirmed that both ggpps and ts gene were stably integrated into the genome of C. cinerea. Crude extracts from each of the transformants were analyzed. There is no difference in the mycelium extracts among the wild-type LT2 and two types of transformants. However, analysis of culture filtrates indicated that an additional GC peak was found at the retention time of 16.762 min which was absent in the wild type control. The mass fragmentation pattern of this peak had the same diagnostic ions with taxa-4(5),11(12)-diene. According to peak area, the amounts of taxa-4(5),11(12)-diene in each fermented broth were 44 ng/L (transformed with pBgGGgTS) and 30 ng/L (transformed with pBgGGTS), respectively. In conclusion, co-expression of the ggpps and ts gene could increase the taxadiene production in C. cinerea.
Agaricales ; metabolism ; Alkenes ; metabolism ; Diterpenes ; metabolism ; Farnesyltranstransferase ; genetics ; metabolism ; Genetic Engineering ; Isomerases ; genetics ; metabolism ; Paclitaxel ; Plasmids