The content of benzyl isothiocyanate (BITC) which as the enzymatic hydrolysis product of benzyl glucosinolate through thioglucosidase was determined by HPLC. The content of benzyl isothiocyanate (BITC) which as the enzymatic hydrolysis product of benzyl glucosinolate through thioglucosidase was determined by HPLC. The chromatography condition was as follows: Kaseisorb LC ODS 2000 (4.6 mm x 150 mm, 5 min) column with the mobile phase of acetonitrile(A)-water( B) under gradient elution (0-5 min, 3%-8% A; 5-9 min, 8%-48% A; 9-23 min, 48%-62% A; 23-28 min, 62%-99% A); the flow rate was 1.0 mL x min(-1) with 10 microL injection volume; detection wavelength was 246 nm and temperature of column was 40 degrees C. The content of benzyl glucosinolate was in the range of 10.76-17.91 g x L(-1). The method is simple, accurate and good reproducibility which can be used for the determination of benzyl glucosinolate in Lepidium meyenii, effectively.
Chromatography, High Pressure Liquid ; methods ; Glucosinolates ; analysis ; Lepidium ; chemistry ; Plant Extracts ; analysis

Objective:To retrospectively assess the relationship between immune disorder and acute gastrointestinal injury (AGI) in patients after severe polytrauma.Methods:Totally 205 patients with severe polytrauma admitted to Tongji Hospital from April 2018 to October 2019 were enrolled as the observation group, and 23 healthy volunteers were served as the control group. According to the diagnostic criteria of AGI, all patients were divided into the AGI group (with AGI) or N-AGI group (without AGI), AGI patients were divided into the S-AGI group or L-AGI group according to the severity. The levels of cytokines and lymphocyte subset were evaluated at day 1, 7, and 14 after severe polytrauma. The differences between groups were statistically analyzed. The independent risk factors of AGI were analyzed by Logistic regression analyzed.Results:Totally 79.5% (163/205) of patients with severe polytrauma were accompanied by AGI. There were significant differences in the ratio of Tc, Th at day 1 after trauma, the levels of IL-6, TNF-α, IL-8, IL-10, the ratio of Ts, Th/Ts, Treg at day 7 after trauma, and the levels of IL-8, IL-10,the ratio of Ts, Th/Ts, Treg at day 14 after trauma between the AGI group and N-AGI group ( P<0.05). There were significant differences in the ratio of Tc, Th, the levels of IL-6, TNF-α at day 1 after trauma and the ratio of Ts, Th/Ts, Treg, the levels of IL-8, IL-10 at day 7 and 14 after trauma between the S-AGI group and L-AGI group ( P<0.05). Logistic regression analysis showed that Ts 7 d ( OR=2.018, 95% CI: 1.105-5.364, P=0.013), Treg 14 d ( OR=3.612, 95% CI: 1.375-8.476, P=0.006), IL-6 7 d ( OR=1.824, 95% CI: 1.011-5.835, P=0.024), IL-10 14 d ( OR=2.847, 95% CI: 1.241-6.216, P=0.014), TNF-α 7 d ( OR=1.754, 95% CI: 1.215-5.441, P=0.018) were independent risk factors in patients with AGI after severe polytrauma. Conclusions:AGI is more easily occurred in patients with the heavier immune disorders after severe polytrauma. AGI can also aggravate pre-existing immune disorders in patients after severe polytrauma.

OBJECTIVETo study the antiproliferation effect on HepG2 cells and its underlying mechanism of the active chemical composition of the Viburnum Odoratissimum.

METHODS3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay and trypan blue dye exclusion assay were used to assess the effect of vibsane-type diterpenoids on the proliferation of various tumor cells. Alterations in cell cycle and apoptosis were determined by flowcytometry. The enzymatic activity of caspase-3/7 was measured by Apo-ONE homogeneous Caspase-3/7 Assay kit.

RESULTSCompound 1 #, a vibsane-type diterpenoid, was found to significantly inhibit the growth of HepG2 cells by anticancer proliferation activity screening. It was demonstrated that the modified groups on side chain coupled to C11 site affected the cell growth-inhibition activity of compounds by structure-activity analysis. In addition, HepG2 cell line was most sensitive to compound 1 #, which induced growth arrest of HepG2 cells in a dose- and time-dependent manner. Study on the mechanisms underlying these effects indicated that compound 1 # induced significant G0/G1 phase arrest of HepG2 cells in a time- and concentration-dependent manner. Meanwhile, It was found that higher concentrations of compound (5-10 micromol/L) caused evident increase in the unmber of apoptotic cells and dose-dependent activation of caspase-3/7.

CONCLUSIONVibsane-type diterpenoids could significantly inhibit the growth of HCC HepG2 cells. Induction of cell cycle arrest and apoptosis may play important roles in their anticancer effects.

Apoptosis ; drug effects ; Cell Cycle Checkpoints ; drug effects ; Cell Proliferation ; drug effects ; Diterpenes ; pharmacology ; Hep G2 Cells ; Humans ; Viburnum ; chemistry

OBJECTIVETo examine the influence of vascular endothelial growth factors (VEGF) in controlling the growth of an experimental osteosarcoma in mice by performing retrovirus-mediated sFlt-1 gene modification.

METHODSFrom March to October 2010 human osteosarcoma G-292 cells were in vitro infected with retroviral vectors encoding soluble Flt-1 or LacZ gene before transplanted into proximal tibiae of immune deficient SCID mice to establish experimental orthotopic osteosarcoma. Daily observation and biweekly microCT were performed to monitor tumor development and progression till sacrifice at 8 weeks after tumor cell inoculation for histological and molecular analyses.

RESULTSSuccessful transgene expression was confirmed in the culture media of sFlt-1 transduced G-292 cells using ELISA, and with positive X-gal staining of the LacZ transduced cells. Noteworthy tumors were grown in all mice on the tibiae receiving G-292 cell inoculation, with clear detection on microCT images starting 2 weeks after inoculation. Over the time period, tumors derived from sFlt-1 transduced G-292 cells were distinctively smaller in size compared to the ones from wide-type G-292 and G-292-LacZ cells. Histology showed typical osteosarcoma characteristics including severe cellular pleomorphism, bone erosions, and neo-vascularization. Real-time polymerase chain reaction indicated significantly higher sFlt-1 expression in sFlt-1 transduced groups than the wild-type G-292 or LacZ treated groups. Strong expression of oncogenes c-myc and c-fos were also obvious, along with the expression of VEGF in the primary tumor tissue.

CONCLUSIONRetrovirus-mediated sFLT-1 gene modification decelerates the osteosarcoma tumor growth in this murine model.

Animals ; Bone Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Female ; Genetic Vectors ; Humans ; Lac Operon ; Mice ; Mice, SCID ; Neovascularization, Pathologic ; metabolism ; pathology ; Osteosarcoma ; genetics ; metabolism ; pathology ; Retroviridae ; genetics ; Transgenes ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism

OBJECTIVETo compare the differences in cerebral oxygenation responses between the infants born preterm and full-term infants and to evaluate the early cognitive ability of infants born preterm.

METHODSCerebral oxygenation after light stimulation was detected by near infrared spectroscopy (NIRS) in preterm infants at 3 or 6 months corrected gestational age (GA). The results were compared with those of age-matched infants born at term.

RESULTSThe start and peak response time of cerebral oxygenation occurring after light stimulation in preterm infants at 3 months corrected GA was 17.2 +/- 5.2 and 38.4 +/- 9.6 seconds respectively, which were significantly longer than in age-matched term infants (13.1 +/- 2.7 and 28.9 +/- 5.0 seconds respectively) (P < 0.05). The maximum response value of hemoglobin, oxyhemoglobin and regional oxygen saturation of the preterm infants at 3 months corrected GA was (1.2 +/- 0.5)%, (1.5 +/- 0.6)%, and (1.3 +/- 0.4)% respectively , which were significantly lower than that of the term infants [(2.3 +/- 0.3)%, (2.8 +/- 0.3)% and (2.4 +/- 0.5)% respectively] (P < 0.05). Cerebral oxygenation responses to light stimulation in preterm infants examined at 6 months corrected GA were not significantly different from age-matched term infants.

CONCLUSIONSCerebral oxygenation responses to light stimulation in infants born preterm at 3 months corrected GA are not as good as age-matched term infants, but were close to the level of age-matched term infants at 6 months corrected GA. This suggests that the early cognitive ability of preterm infants before 3 months corrected GA might fall behind age-matched term infants.

Brain ; metabolism ; Cognition ; Humans ; Infant, Newborn ; Infant, Premature ; Oxygen ; metabolism ; Photic Stimulation ; Spectroscopy, Near-Infrared

Objective To investigate the effects of NADPH oxidase inhibition on cardiac function and myocardial calcium regulatory proteins mRNA expressions in rabbits with heart failure (HF). Methods HF was induced by experimental aortic insufficiency and abdominal aortic constriction, HF animals were treated with oral apocynin (15 mg/d), a NADPH oxidase inhibitor or equal dose placebo. Eight weeks later, cardiac function was measured by echocardiography. Myocardial NADPH oxidase activity was evaluated by NADPH dependent superoxide production examined using superoxide dismutase-inhibitable cytochrome e reduction. Sarcoplasmic reticulum Ca~(2+) ATPase (SERCA2a), ryanodine receptor 2 (RyR2), phespholamban (PLB) and sedium-calcium exchanger (NCX) were determined by RT-PCR. Results Rabbits with HF developed ventricular dilatation and cardiac dysfunction, as well as increase in myocardial NADPH oxidaoe activity, decreases in mRNA expression of SERCA2a, RyPt2 and PLB, and increase in mRNA expression of NCX. Apocynin significandy reduced NADPH exidaae activity (P<0.05), upregulated SERCA2a, RyP,2 and PLB mRNA expressions (SERCA2a/GAPDH:0.63±0.11 vs. 0.34±0.08,RyR2/GAPDH:0.23±0.04 vs. 0.17±0.06,PLB/GAPDH:1.28±0. 13 vs. 0.95±0.09, P <0.05 ), downregulated NCX mRNA expression ( NCX/GAPDH :0.67±0.10 vs. 0.95±0.12, P<0.05 ), and improved cardiac function [ LVEF: (60.06±10.07) % vs. (38.87±3.31) %, LVFS: (30.12± 6.56)% vs. (17.40±2.45)%,P<0.05] in rabbits with HF. Conclusion NADPH oxidase inhibition improves cardiac function possibly by preventing abnormal alterations in myocardial calcium regulatory proteins in failing heart.

Objective:To prospectively assess clinical characteristics, potential causes and prognosis in patients with persistent inflammation, immunosuppression and catabolism syndrome (PICS) after polytrauma.Methods:Totally 1 083 patients with polytrauma admitted to Department of Traumatic Surgery of Tongji Hospital from Janury 2019 to July 2020 were enrolled. Exclusion criteria included age<18 years old, length of hospital stay<15 days, previous medical history of malignancy, or immunological, consumptive, and metabolic diseases. According to the diagnostic criteria of PICS, all enrolled patients were divided into two groups: PICS group and N-PICS group (without PICS). The patient’s clinical characteristics, ISS score, GCS score, SOFA score, and prognosis were collected. The χ2 test or Student’s t test was uesd to compare the difference between the PICS group and N-PICS group. Results:The incidence of PICS in patients with polytrauma was 11.7% (127/1 083). The majority of PICS patients were middle-aged and elderly men, 68.5% with traumatic brain injury and 59% with thoracic injury. GCS score was significantly lower, while ISS, APACHE II and SOFA scores were significantly higher in the PICS group than in the N-PICS group ( P<0.01, P<0.05). Among PICS patients, 79.5% were treated with mechanical ventilation and 76.3% were associated with pulmonary infection, with a 28-day mortality of 5.5% and a 180-day mortality of 16.5%, which were siginifcantly different from those without PICS. Conclusions:PICS has a high incidence after polytrauma and is commonly seen in middle-aged and elderly male patients with severe polytrauma, especially accompanied by traumatic brain injury or/and thoracic injury. Patients with PICS after polytrauma have poor long-term prognosis, so early identification and intervention should be strengthened in clinical practice.

OBJECTIVETo study correlation of brain hypoxia of different degrees with brain function and damage.

METHODSThe brain regional oxygen saturation (rSO2) was determined by using a non-invasive near infrared spectroscopy (NIRS) technique in 15 piglets; the piglets were subjected to inhale 3% - 11% oxygen-nitrogen mixed gas through mechanical ventilation for 30 min. The piglets were divided into groups according to the level of brain rSO2 (i.e. < 30%, 30% - 35%, 35% - 40%, and 40% - 50%), and the data were compared with those of the control group (rSO2 > 60%). Changes of brain function were detected through amplitude and frequency of EEG waves and signal complexity. The piglets were sacrificed via decapitation 72 h after brain damage, and then histopathological and ultrastructural examinations were performed on cerebral cortex and hippocampal CA1 area.

RESULTSIn the group with rSO2 > 40%, the mean arterial pressure (MAP) after hypoxia was (56 +/- 0.00) mm Hg (1 mm Hg = 0.133 kPa), the blood lactic acid (LA) was (2.3 +/- 1.2) mmol/L, the EEG findings were within normal range, and there was no change in brain tissue ultrastructure. In the group with brain rSO2 = 30% approximately 40%, the MAP was (73 +/- 8) mm Hg, the LA was (8.2 +/- 3.9) mmol/L, the EEG waves showed decreased amplitude, frequency and complexity, but restored to some extent after hypoxia. The brain tissue ultrastructure showed damages to the cerebral cortex and neuron mitochondria at hippocampal CA1 area. In the group with brain rSO2 < 30%, the MAP was (35 +/- 0) mm Hg, the LA was (12 +/- 2) mmol/L, the EEG showed decreased amplitude, frequency, and complexity of signals compared with those of the normal control group, and was difficult to restore after hypoxia in some of the piglets; the brain tissue ultrastructure appeared to be similar to the changes seen with high-degree swollen cerebral cortex and neuron mitochondria at hippocampal CA1 area.

CONCLUSIONDifferent degrees of hypoxia had different influence on brain function and brain damage. The lower the brain rSO2, the more severe the damages to the brain and its function. The rSO2 of brain tissues detected with noninvasive NIRS can reflect brain injury and its severity during cerebral anoxia.

Animals ; Blood Gas Analysis ; Brain Injuries ; complications ; Cerebral Cortex ; physiopathology ; Cerebrovascular Circulation ; physiology ; Electroencephalography ; Female ; Hypoxia ; metabolism ; pathology ; Hypoxia, Brain ; complications ; Hypoxia-Ischemia, Brain ; physiopathology ; Male ; Neurons ; pathology ; Oximetry ; instrumentation ; Oxygen ; metabolism ; Oxygen Consumption ; Spectroscopy, Near-Infrared ; methods ; Statistics as Topic ; Swine

Objective:To retrospectively assess the occurrence and risk factors in patients with acute gastrointestinal injury (AGI) after polytrauma.Methods:Totally 430 patients with polytrauma admitted to Tongji Hospital from April 2018 to October 2019 were enrolled as the observation group. According to the diagnostic criteria of AGI, all patients were divided into the AGI group (with AGI) or N-AGI group (without AGI). The patients with abdominal injury or previously suffered from gastrointestinal disease were excluded. The patient's clinical characteristics lab tests results, and the first ISS, APACHEⅡ, SOFA and GCS scores were collected. The differences between different groups were statistically analyzed. The independent risk factors of AGI were analyzed by Logistic regression.Results:65.3% of patients with polytrauma were accompanied by AGI (281/430 cases).There were significant differences between the AGI group and N-AGI group in ISS, GCS, APACHE Ⅱ and SOFA score, PCT or IL-6 level, shock index and length of stay in ICU ( P<0.05). Logistic regression analysis showed that shock, ISS≥16, APACHE Ⅱ≥16, SOFA≥5, GCS≤8 and IL-6>50 pg/mL were the early independent risk factors in patients with ACI after polytrauma. Conclusion:The incidence of AGI in patients after polytrauma is higher, which is related to ischemia, hypoxia, abnormal blood coagulation and stress in the early stage after trauma.

OBJECTIVETo find an approach for trans-oral endoscopic thyroidectomy (TOET) and cervical lymphadenectomy using conventional endoscopic surgical instruments on frozen fresh cadavers.

METHODSSix frozen fresh cadavers were used in three groups of trans-oral trocar installation experiments: oral vestibule installation, sublingual region installation, and combined bi-vestibular and sublingual installation. TOET (with pretrachealis method to thyroid fixation removal) and cervical lymphadenectomy were performed experiments on another 6 frozen fresh cadavers using the best access approach found in the aforementioned experiments.

RESULTSIn oral vestibule trocar installations, the trocars caused large lacerated wound and damaged air tightness. In sublingual installations, only one trocar could be installed in the sublingual area because the space in sublingual area was limited. In combined bi-vestibular and sublingual installations, no gland, vessel or nerve was damaged. Combined bi-vestibular and sublingual access were selected as the surgical approach on the basic of analysis the merits of each approach. TOET and cervical lymphadenectomy in area III, IV, VI, VII were performed without making any accessory damage through combined bi-vestibular and sublingual access approach.

CONCLUSIONSTOET is feasible. Combined bi-vestibular and sublingual approach is available for TOET. Part of the cervical lymph nodes could be resected. Pretrachealis approach to thyroid fixation removal can still be used.

Adult ; Cadaver ; Endoscopy ; Humans ; Lymph Node Excision ; methods ; Neck ; Thyroidectomy ; methods