AIM: To investigate the possibility that hepatocyte growth factor (HGF) directly induces differentiation of human embryonic stem cells (hESCs) into neural progenitors (NPs). METHODS: hESCs colonies were induced to form the embryoid body (EB). Four-day-old EBs were randomly divided into 4 groups: control group (EBs were cultured in neural induction medium);G5 supplement group (EBs were cultured in neural induction medium supplied with G5 supplement);HGF group (EBs were cultured in neural induction medium supplied with 10 mg/L HGF), and HGF+G5 group (EBs were cultured in neural induction medium supplied with 10 mg/L HGF and G5 supplement). After induced in suspension system for 7 days, EBs with various treatments were cultured in poly-D-lysine/laminin-coated plates for 7-10 days for selection of NPs. NPs were gathered by 0.3 g/L dispase treatment and characterized by immunofluorescence staining. The percentages of the nestin+ cells in NPs in various groups were detected by fluorescent activated cell sorter (FACS). The multipotency of NPs was determined by immunofluorescence staining after the NPs were cultured without G5 and HGF for 7 days. The expression of region markers of neural progenitors treated with sonic hedgehog (Shh) protein (one of the neural inductive signals), was detected by RT-PCR. RESULTS: HGF+G5 supplement induced hESCs differentiation into neural progenitors. Immunofluorescence staining indicated that NPs differentiated from hESCs expressed NP markers including nestin, Pax6 and musashi-1. FACS data showed that the proportion of nestin positive cells in HGF+G5 supplement group (87.3%±3.9%) was the highest in all treatment groups. The time of HGF and G5 supplement treatment was important to differentiate into NPs, the maximal effect was observed at 7th day. After treated with Shh, the expression of ventral forebrain/hindbrain marker genes (Nkk2.1, and Nkk2.2) and hindbrain progenitor marker gene Gbx2 in NPs were upregulated, while the forebrain progenitor marker genes Otx2 and Bf1 were downregulated. CONCLUSION: The neural induction system containing HGF and G5 supplement effectively induces the differentiation of hESCs into NPs, which might be a potent model for investigating the mechanism of neural development and differentiation.

Objective To investigate if mitochondrial KATP channels are involved in the cardioprotective effects of astragalus membranaceus in immature rabbit hearts. Methods Twenty-four New Zealand white rabbits of both sexes aged 14-21 days weighing 300-350 g were anesthetized and heparinized. The hearts were rapidly removed after thoracotomy and mounted on a Langendorff apparatus via ascending aorta and perfused with oxygenated (95% O2-5% CO2) Krebs-Hensleit buffer (KHB) solution at 60 cm H2O (perfusion pressure) and 38 ℃ . A fluid-filled latex balloon was inserted into left ventricle via left atrium for measurement of left ventricular developed pressure ( LVDP) , Global myocardial ischemia was induced by suspension of perfusion for 30 min followed by 45 min reperfusion. The animals were randomly divided into 3 groups with 8 animals in each group : group A control;group B astragalus and group C astragalus +- 5-HD (a selective inhibitor of the mitochondrial KATP channel). In group B the hearts were perfused with astragalus 40 g? L-1 for 15 min before ischemia. In group C the hearts were perfused with 5-HD 100?mol?L-1 for 5 min followed by 10 min astragalus perfusion before ischemia. In control group the hearts were perfused with only KHB before ischemia. Coronary flow (CF), HR, LVDP and ? dp/dtmax, were measured at 5, 10, 15, 30 and 45 min of reperfusion and recorded. Coronary effluent was collected at 10 min of reperfusion for determination of CK-MB, LDH and CK levels . At the end of 45 min reperfusion the isolated heart was removed for determination of myocardial ATP and iNOS levels and electron microscopic examination.Results In astragalus group (B) LVDP ? dp/dtmax and CF recovered significantly better and myocardial ATP content was significantly higher as compared with group A and C (P

Objective To establish a simple ,stable acute liver injury model induced by CCl4 to observe effects of hepatocyte transplantation .Methods CCl4 plant oil with different concentration of 20% and 50% was used in mice by intraperitoneal injection , of which the dose was 2 mL/kg ,and then materials were taken at different time points respectively .Mice survival rate ,alanine amin-otransferase (ALT) ,aspartate aminotransferase (AST) and the pathological changes of the liver were detected .Results Mice sur-vival rate in 20% CCl4 intraperitoneal injection was significantly higher than that of 50% .ALT and AST in experiment group were significantly higher than that of control group ,but there was no significant difference between two experimental groups .Pathologi-cal examination showed that mice liver cells showed typical cytoplasmic ,ballooning ,scattered punctate ,piecemeal necrosis and in-flammatory cell infiltration in 20% CCl4 intraperitoneal injection ;while in 50% CCl4 ,there was obvious fibrosis ,in addition to the mentioned heavier lesions .Conclusion 20% -50% CCl4 intraperitoneal injection in 2 mL/kg dose can induce different degrees of relatively stable liver injury ,and its concentration determines the degree of liver injury .Acute liver injury induced by 20% -50%CCl4 was an ideal model for hepatocyte transplantation experiment .

BACKGROUND:FGF2,TGFβ/activin/nodal and IGF signal pathways are necessary for keeping functions of human embryonic stem calls,but there was no report concerning whether addition of exogenous basic fibroblast growth factor,transforming growth factor β and insulin can maintain self-renewal of human embryonic stern cells.OBJECTIVE:To establish a feeder layer-and serum-free culture system of human embryonic stem cells.DESIGN,TIME AND SETTING:The cytological in vitro study was performed at the Kunming Animal Institute of Chinese Academy of Sciences from September 2007 to February 2009.MATERIALS:Pregnancy 12.5 or 13.5-day ICR strain mica (clean grade) were provided by the Animal Center of Kunming Medical College.Human embryonic stem cell line BG02 was purchased from Bresagen,USA.METHODS:BG02 cells were digested,centrifuged,resuspended in feeder layer-free medium,and then incubated for 5-7 days.Differentiated human embryonic stem cells were removed.Dispase was added for digestion.Samples were cut into blocks,centrifuged,resuspended,and then incubated at 1:3 in a 4-well plate coated with laminin in feeder layer-free and serum-free medium,supplemented with 80% DMEM/F12,20% KSR,2 mmol/L glutamine,1% non-essential amino acid,0.1 mmol/L β-mercaptoethanol,insulin-transferrin-selenium (ITS) (×1),10~6 U/L penicillin,100 mg/L streptomycin,4 μg/L basic fibroblast growth factor (bFGF) and 0.12 μg/L transforming growth factor-β1 (TGF-β1).ICR fetus mica were sterilely obtained to culture mouse embryonic flbroblasts by tissue pancreatin digestion method.These cells were incubated in a 6-well plate coated with 0.1% gelatin.Human embryonic stem cell line BG02 cultured on mouse embryonic fibroblasts feeder layers was transferred to laminin-coated plates in serum-free medium containing bFGF,TGFβ1 and ITS.MAIN OUTCOME MEASURES:The morphology of BG02 cells in feeder layer-and serum-free condition was observed.The specific molecular markers of human embryonic stem cells were determined by immunohistochemical staining.The karyotype and differentiation ability in vitro of BG02 cells were analyzed.The differences in the proliferation and the differentiated rate of BG02 clumps in feeder layer-and serum-free condition or mouse embryonic fibroblast feeder layer condition were compared.RESULTS:BG02 calls had been continuously cultured for at least 20 passages in feeder layer-and serum-free culture condition.BG02 clones in this culture system had typical human embryonic stem cell morphology.BG02 cells all expressed SSEA-4,SSEA-3,TRA-1-60,TRA-1-81,Oct-4,but did not express SSEA-1.After 20 days of culture,BG02 cells had the capacity to differentiate into derivatives of embryonic endoderm,mesoderm,and ectoderm.Differentiated cells could express alpha-fetoprotein,nestin,α-actin.At passage 20,call karyotype was normal (46XY).The growth of BG02 cells cultured in feeder layer-and serum-free condition was more slowly compared with mouse embryonic fibroblast feeder,and doubling time was significantly prolonged (P ＜ 0.05).Differentiation rate of the colonies was significantly increased (P ＜ 0.05).CONCLUSION:A feeder layer-and serum-free condition for culture of BG02 cells has been established.The addition of bFGF,TGFβ1 and ITS can maintain the self-renewal of BG02 cells.

Objective To investigate the protective effect of hepatocyte growth factor (HGF) on cultured Sprague-Dawley rat cortical neurons injured through hypoxia/reoxygenation.Methods Primary cultured cerebral cortical neurons were isolated from newborn rots.Neurons were pre-incubated with different concentrations (15,30 and 60 μg/L) of HGF.The cell viability was detected by MTT.Apoptosis was measured by Hoechst 33258 staining and flow cytometer.Lactate dehydrogenase (LDH) and caspase-3 activity were determined by colorimetry.Results Compared with normal group,hypoxia/reoxygenation treatment significantly decreased cell viability,increased LDH activity and the percentage of apoptotic cells.Pretreatment of HGF for 12 h could remarkably reverse the decrease of cell viability and the increase of apoptosis rate in neurons induced by hypoxia/reoxygenation treatment.HGF pre-treatment also attenuated the activity of LDH and caspase-3 in a dose-dependent manner.The effects of HGF could be inhibited by a special PI3K/Akt pathway inhibitor,LY294002.Condusion HGF could attenuate rat cortical neuron injury induced by hypoxia/reoxygenation.The neuroprotective effect of HGF may be related to activating PI3K/Akt pathway,and further suppressing the expression of caspase-3.

OBJECTIVE To investigate the clinical characteristics and drug resistance of meticillin-resistant Staphylococcus aureus(MRSA) among inpatients in Huaxi Hospital in five years.METHODS We analyzed clinical distribution and resistant data on inpatients infected with MRSA between Jan 2003 and Dec 2007 in Huaxi Hospital,and collected the annual consumption of common used antibiotics,amount of MRSA isolates and MRSA isolated rate from Staphylococcus aureus(SA) strains.The data were analyzed by the method of Spearman analysis in SPSS.RESULTS We analyzed totally 1478 MRSA strains.The most common specimens of MRSA were sputum(70.8%),other secretions(14.2%) and blood(4.6%).The MRSA strains were frequently detected in ICU(29.8%),Department of surgery(17.3%) and Department of respiratory Medicine(6.4%).The drug resistance rate to ampicillin,tetracycline,rifampin,imipenem,ciprofloxacin,gentamicin,cefalothin,cefotaxime and cefazolin were beyond 90% in five years.The drug resistance rate against Erythromycin,Clindamycin and Clarithromycin decreased gradually from 93.1%,92.2% and 95.5% in 2003 to 82.3%,76.7% and 81% in 2007.It had not been found the vancomycin-resistant MRSA.Except Cotrimoxazole,the drug resistance rates of MRSA in ICU were almost higher than those in non-ICU departments.Based on the SPSS analysis,we concludea that annual consumption of etimicin had a positive correlation with MRSA,and

BACKGROUND:There is evidence that internal fixation through an anterior or posterior approach for treatment of severe kyphotic deformity in spinal tuberculosis exhibits good curative effects. However, few prospective, long-term fol ow-up case control studies are reported. OBJECTIVE:To investigate the efficacy and safety of cobalt al oy pedicle screw implantation for treatment of severe kyphotic deformity in spinal tuberculosis. METHODS/DESIGN:This is a prospective, single-center, self-control, open-label trial, which wil be performed at the Affiliated Hospital of Hebei University, China. Eighty-four patients with severe kyphotic deformity in spinal tuberculosis wil be included according to the diagnosis criteria. Among 52 patients with tuberculosis of the thoracic spine, 28 wil undergo surgery through a posterior approach, and 24 through an anterior approach. According to the Frankel Grade classification, grade C, D and E spinal cord function wil be assessed in 8, 31 and 13 patients, respectively. Surgery through a posterior and anterior approach wil be respectively performed in half of 32 patients with tuberculosis of the lumbar spine. Grade C, D and E spinal cord function wil be assessed in 7, 14 and 11 patients, respectively. The primary outcome measure of this study wil be the Cobb angle at the thoracic spine segments before and 2 years after surgery, which wil be used to evaluate the angle of the spine curvature at the thoracic segments. The secondary outcome measures wil be X-ray scan or MRI findings before and 2 years after surgery, which wil be used to evaluate vertebral fusion after internal fixation;and Frankel Grade before and 2 years after surgery, which wil be used to evaluate recovery of spinal cord function after injury. Other outcome measures wil include multiple logistic regression analysis results of the factors that influence patient's curative effects and the incidence of adverse events 2 years after surgery. The trial protocol has been approved by the Ethics Committee, Affiliated Hospital of Hebei University, China and wil be performed in strict accordance with the Declaration of Helsinki, formulated by the World Medical Association. Signed informed consent regarding the trial protocol wil be obtained from each participant. DISCUSSION:This study is to validate that cobalt al oy pedicle screw implantation shows precise curative effects in the treatment of severe kyphotic deformity in spinal tuberculosis and to analyze through what approach, posterior or anterior, internal fixation wil be more beneficial to surgery performance. The outcomes of this study wil provide objective long-term fol ow-up evidence for internal fixation treatment of severe kyphotic deformity in spinal tuberculosis in the clinic.

BACKGROUND: During the repair of thoracolumbar fracture, pedicle screw fixation is a commonly used treatment method. In the process of fixation, the different approaches can be used. OBJECTIVE: To compare effect and biocompatibility of pedicle screw by percutaneous approach, posterior median approach, and intervertebral space approach for thoracolumbar fracture. METHODS: 118 cases of thoracolumbar fracture were included after pedicle screw fixation. Al patients were divided into three groups according to the approach: posterior median approach group (38 cases), intervertebral space approach group (40 cases) and percutaneous approach group (40 cases). After 12 months of folow-up, perioperative conditions, pain score, vertebral height of anterior border, kyphosis correction effect, adverse events and biological compatibility were compared among three groups. RESULTS AND CONCLUSION:(1) Operation time, intraoperative bleeding and time in bed after surgery were shorter or less in the percutaneous approach and intervertebral space approach groups than in the posterior median approach group. Postoperative drainage was better in percutaneous approach and intervertebral space approach groups than in the posterior median approach group (alP < 0.05). Except drainage in the percutaneous approach and intervertebral space approach groups, no significant difference in other indicators was found. (2) Patients received imaging examination at different time points. The percentage of anterior vertebral height and kyphosis were significantly improved immediately after treatment and in final folow-up (alP < 0.05). No significant difference was detected before treatment, immediately after treatment and in final folow-up. (3) Visual Analogue score was identical before treatment. Visual analogue score was lower in the percutaneous approach and intervertebral space approach groups than in the posterior median approach group at 24 hours and 3 days after treatment and in final folow-up (alP < 0.05). No significant difference was detectable at 24 hours and 3 days after treatment and in final folow-up between the percutaneous approach and intervertebral space approach groups. (4) No rejection or wound non-healing was seen at 12 months after treatment. Some patients suffered from mild low back pain, which was improved by active symptomatic treatment. (5) These findings suggest that intervertebral space approach percutaneous approach obtained satisfactory outcomes compared with posterior median approach for treatment of thoracolumbar spine fractures, and good biocompatibility was found.

OBJECTIVETo observe the liver injury degree of SD rats after 28-day administration of aqueous extracts of Polygonum multiflorum (AEPM) and the correlation with cholestasis mechanism.

METHODAdult SD rats were orally administered with 30, 60 g x kg(-1) of APEM once every day for 28 d. After 28 d, the general condition of rats such as weight were observed, liver function-related indicators were detected. Bile was collected to determine total bile acid output, flow rate and density and changes in major compositions. Their livers were weighed then sent for histopathological examination.

RESULTAEPM did not change the general conditions and weights of rats. From the results of the related indicators of liver function and cholestasis, AEPM did not change the contents of ALT and AST in serum, but high dose of AEPM can increase the contents of ALP, GGT and TBA in serum (P < 0.05, P < 0.01) and decrease the content of TBIL in serum (P < 0.05). And the contents of GGT in serum of low dose rats were increased (P < 0.05). The bile flow was not changed by AEPM, but bile compositions of high dose male rats were obviously changed (TG increase, TBIL decrease, TBA decrease). The weights of liver and ratio of liver of the high dose rats were increased but showed no statistical significance. Pathologic examination displayed that there were only small pieces of necrosis in livers of several rats, without any severe disease.

CONCLUSIONAEPM can obviously injure bile duct epithelial cells, intervene liver cell functions and change bile compositions in rats, thus it is proved to induce cholestasis without severe liver injury.

Administration, Oral ; Animals ; Bile ; chemistry ; drug effects ; Cholestasis ; chemically induced ; Female ; Liver ; drug effects ; pathology ; Male ; Plant Extracts ; toxicity ; Polygonum ; toxicity ; Rats ; Rats, Sprague-Dawley

Objective:To compare four reconstruction algorithms of 18F-fluorodeoxyglucose (FDG) PET/CT on standardized uptake value (SUV) of pulmonary nodules. Methods:A total of 46 patients (27 males, 19 females; median age: 66 (range: 44-82) years) with solid pulmonary nodules from February 2018 to July 2019 in the First Hospital of Shanxi Medical University who performed 18F-FDG PET/CT imaging were enrolled. All PET/CT images were retrospectively reconstructed by using four algorithms reconstructions including ordered subset expectation maximization (OSEM), OSEM+ time of flight (TOF), OSEM+ TOF+ point spread function (PSF) and block sequential regularized expectation maximization (BSREM) (G1-G4). Nodule and background parameters were analyzed semi-quantitatively and visually. The maximum of SUV(SUV max), mean of SUV(SUV mean) and peak of SUV (SUV peak) were collected by the region of interest (ROI). Nodules were divided into small nodule group (diameter ≤10 mm) and large nodule group (10 mm < diameter ≤30 mm). Kruskal-Wallis rank sum test and Bonferroni method were performed to compare the differences of SUVs between G1-G4, and Spearman correlation analysis was used to analyze the correlation between the change rate of SUV (%ΔSUV) and the diameter of nodules. The receiver operating characteristic (ROC) curve analysis was used to analyze the diagnostic efficacy of SUV for the differential diagnosis of pulmonary nodules and to get the optimal threshold. Results:There were 114 pulmonary nodules (large nodules, n=55; small nodules, n=59). In visual analysis, the visual detection rates of small nodules in G4 were 55.93%(33/59), 44.07%(26/59), 20.34%(12/59) higher than those in G1-G3. Of 114 pulmonary nodules in 46 patients, there were differences in SUV max and SUV mean between G1-G4 (median SUV max : 2.65-5.29, median SUV mean: 2.05-2.99; H values: 20.628 and 17.749, respectively, both P<0.001), G4 had significant increases compared to G1 in SUV max (median 5.29 and 2.65, P<0.001) and SUV mean (median 2.99 and 2.05, P<0.001). The %ΔSUV max (median: 4.45%-52.96%) and %ΔSUV mean (median: 1.69%-47.56%) were negatively correlated with the diameter of nodules (9.75(6.20, 16.58) mm; r s values: -0.371 to -0.354, -0.371 to -0.320, all P<0.001). In 59 small nodules, G1 significantly increased the SUV max of G4 (median 4.05 and 2.14, H=18.327, P<0.001), while G4 significantly increased the SUV mean of G1 and G3 (median 2.31, 1.26 and 1.53, H=16.808, P<0.05). There was no significant difference in SUVs between G1-G4 in 55 large nodules ( H values: 0.812-7.290, all P>0.05). The optimal threshold values of SUV max in G1-G4 were 4.335, 5.185, 5.410, 5.745 and the area of under curves (AUCs) were 0.747, 0.699, 0.756, 0.778 respectively. The AUC of SUV mean and SUV peak also showed a similar trend. Conclusion:Among the four reconstruction algorithms, BRERM can not only enhance the image quality, but also significantly improve the SUV max and SUV mean of lung nodules diameter below 10 mm, and thus its diagnostic threshold of SUV should be appropriately increased.