Objective To explore the therapeutic effect and application value of the self-made spinal positioning equipment and percutaneous pedicle screw fixation system in the treatment of thoracic and lumbar fracture. Methods To determine the fractured vertebra and pedicle screws entrance point with spinal positioning equipment, 67 patients with thoracic and lumbar fractures were performed percutaneous pedicle screw fixation by using self-made percutaneous pedicle screw fixation system. By contrasting the perioperative indicators, and imaging indicators, to evaluate the therapeutic effect and application value of the system. Results For the spinal positioning equipment, the location time were (15.85±2.45) min and the location accuracy were 95.03%±3.27%. But for the open reduction internal fixation, the location time were (35.46±5.39) min and the location accuracy were 94.02%±2.95%. There was no significant difference in location accuracy, but were significant differences in location time (P＜0.05). Percutaneous pedicle screw fixation had the same effect on three sides with open reduction internal fixation in vertebral height restore,kyphosis deformity and correction and lumbar spinal stenosis's correction. The perioperative indicators of the preoperative and postoperative grope had significant difference (P＜0.05). There were significant differences in all perioperative indicators between the percutaneous pedicle screw fixation and the open reduction internal fixation (P＜0.05). Conclusion The spinal positioning equipment is helpful to determine the fractured vertebra and pedicle screws entrance point accurately and reduce the radiation. The percutaneous pedicle screw fixation system has the advantage of convenient manipulation and accurate implantation. The system can not only reduce surgical damage and post-operation reaction but also make patients recover quickly and face less complications.

Objective To observe changes in the working memory and brain functional imaging on functional magnetic resonance imaging(fMRI) after 36 hours sleep deprivation (SD) in healthy volunteers and to explore the possible mechanism of the changes.Methods FMRI scannings were performed in ten male healthy young volunteers before and after 36 hours SD and results were analyzed using SPM2 software.Subjects were also tested LTR and PLUS task to measure the persistence and operation of working memory before and after 36 hours SD.Results The reaction time of LTR task after 36 hours SD ( (866 ± 102) ms)was significantly longer than that before SD ( (754 ± 91 ) ms, t = 2.59, P < 0.01 ).The reaction time of PLUS task after SD ( (848 ± 94) ms) was significantly longer ( t = 2.37, P < 0.05 ) than that before SD ( (756 ± 79) ms).The error rate of LTR task after SD (95.3% ± 3.56% ) was significantly higher (t=3.52,P < 0.01 ) than that before SD (84.8% ± 8.71% ).The error rate of PLUS task after SD (95.7% ±4.72% ) was significantly higher (t =3.38 ,P <0.01 ) than that before SD (84.2% ±9.66% ).There were no significant differences between the two tasks.The frontal and parietal lobes, anterior cingulate gyrus and thalamus were activated during memory tasks testing before SD.Brain activation was broader and stronger in PLUS task than in LTR task.After SD, activation in parietal lobe was decreased and activation in prefrontal and thalamus was increased significantly.Conclusions The working memory performance decreased after SD.Both LTR and PLUS tasks of working memory activate frontal and parietal lobes, anterior cingulate gyrus and thalamus.The activation of parietal lobe decreased and the activation of prefrontal lobe and thalamus increased after 36 hours SD.This is the possible mechanism of SD to causes the cognition decline.

Objective To investigate the effects of interleukine-22 ( IL-22 ) on the expression of interleukin-6 (IL-6) by rheumatoid arthritis synovial fibroblasts (RASF), and to analyze their association with IL-17+CD4+T (Th17) cells differentiation.Methods RASF were isolated from six patients with rheu-matoid arthritis ( RA) and cultured in vitro.The expression of IL-6 at mRNA and protein levels by RASF were detected by qRT-PCR analysis and ELISA after treatment with different concentrations of IL -22 for dif-ferent periods of time.Anti-IL-22R1 blocking antibody and inhibitor assay were used to analyze the specific receptor and its downstream signaling pathways associated with IL-6 production.IL-22 pre-treated RASF and CD4+T cells were co-cultured for 3 days in the presence or absence of anti-IL-22R1 or anti-IL-6 to measure the percentage of Th 17 cells by flow cytometry .Results The expression of IL-6 by RASF was increased up-on IL-22 stimulation in a dose and time dependent manner (P<0.05), and that was closely related to IL-22R1 and its downstream signaling pathways of p38 and JAK2 (P<0.05).Co-culturing CD4+T cells with RASF and Transwell system indicated that the percentage of Th 17 cells was increased in IL-22 pre-treated group as compared with that in IL-22 untreated group , but it could be down-regulated by either blocking IL-22R1 or IL-6.Conclusion IL-22 promoted the expression of IL-6 by RASF and further enhanced Th 17 dif-ferentiation.Neutralizing IL-22 in synovium of patients with RA might be an effective therapeutic strategy for RA treatment.

AIM: To investigate the function and morphological changes of long-term cultured primary rat hepatocytes. METHODS: Rat primary hepatocytes were isolated by two-step in situ collagenase perfusion method, and then were purified by density and grade centrifugal method with Percoll. Cell viability was observed by 0.4% trypan blue. The hepatocytes were seeded into 6 wells plate with HepatoZYME-SFM medium. ALT, AST, albumin and urea levels in the supernatant were measured, CYPⅠA1 was detected with EROD method. RESULTS: (2-3)?108 cells per whole liver were obtained with viability and purity above 90% after purified with Percoll. Hepatocytes cultured in HepatoZYME-SFM grew well with normal hepatocyte morphometrics. ALT, AST levels in the supernatant decreased after 3-day culture, and kept at a stable level after 6-9 days. Albumin secretion and urea synthesis were maintained at high levels in 18 days, while CYPⅠA 1 enzyme activity was only detected in 3-6 days. CONCLUSIONS: Percoll was used to increase the viability and purity of freshly isolated rat hepatocytes. Hepatocyte morphometrics and their biological synthesized function are effectively maintained in HepatoZYME-SFM medium.

Shenfu injection is a common clinical medicine. To explore the common clinical medical combinations with Shenfu injection the real world, 25 704 patients who used Shenfu injection were selected from the hospital information system (HIS) database established by Clinical and Basic Research Institute for Traditional Chinese Medicine of China Academy of Chinese Medical Sciences. The association rules were applied in a correlation analysis on common medical combinations with Shenfu injection in the patients. According to the findings, when Shenfu injection was combined with a single medicine in clinic, the ratio of using Omeprazole, Lidocaine or Furosemide were respectively 22.19%, 20.32%, 19.61%; when Shenfu injection was combined with two medicines, the top three medical combinations were respectively midazolam + omeprazole (11.01%), lidocaine + omeprazole (10.8%) and propofol + midazolam (10.76%); when Shenfu injection was combined with three medicines, the top three medical combinations were respectively propofol + fentany + midazolam (8.83%), remifentanil + propofol + midazolam (8.77% ) and propofol + midazolam + omeprazole (8.77%). According to the further analysis, the combination of Shenfu injection and Propofol + Fentany + Midazolam may reduce the incidence of hypotension and bradycardia during anesthesia, accelerate recovery after anesthesia and relieve the synergistic effect after anesthesia. The combination of Shenfu injection and furosemide may show the synergistic effect in treating the acute left ventricular failure by reducing the returned blood volume and increasing the myocardial contractile force and vitality. The combination of Shenfu injection and Omeprazole may play the synergistic effect in shortening the digestive tract ulcer healing time, reducing the ulcer recurrence and preventing hemorrhagic shock and stress ulcer caused by shock in treating ulcer hemorrhage.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; Databases, Factual ; Drug Therapy, Combination ; statistics & numerical data ; Drugs, Chinese Herbal ; administration & dosage ; Electronic Health Records ; Female ; Hospital Information Systems ; Humans ; Male ; Middle Aged ; Young Adult

The hexane extract from marine actinomycetes 124092 showed potent inhibition on B16 cell line by MTT assay. The hexane extract was fractionationed on silica gel column by vacuum liquid chromatography to afford 6 fractions(Fr1~Fr6), and Fr6 showed cytotoxic activity. To determine the bioacitve components of hexane extract, Fr6 was analyzed by GC/MS. The main components were identified as palmitic acid (11.76%), oleic acid (12.16%), linoleic acid (14.77%), and lactobacillic acid (61.31%). It have been reported that palmitic acid, oleic acid, and linoleic acid possess cytotoxic activity on rat ascites tumor cells and linoleic acid have suppressive effect on human lung adenocarcinoma cells.

Objective To study the effects of osthole on the proliferation,invasion and migration of nasopharyngeal carcinoma cells CNE2,and to investigate the possible molecular mechanism involved in epithelial to mesenchymal transition (EMT) of CNE2.Methods CNE2 cells were cultured in vitro and were treated with 0,20,40 and 80 μg/ml osthole for 24 or 48 hours,and then methyl thiazolyl tetrazolium (MTT) assay and Transwell assay were used to explore their effects on the cell proliferation,invasion and migration while cells treated with 0 μg/ml osthole were used as the control group.Meanwhile,the mRNA and protein levels of markers of EMT (E-cadherin and vimentin) and Wnt/β-catenin signaling (β-catenin and cyclin D1) were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting respectively.Results After treatment for 24 and 48 hours,the inhibitory rates of treatment with various concentration of osthole (0,20,40,80 μg/ml) were 0.00％ ± 0.00％,7.45％ ± 0.87％,14.12％ ± 2.29％,27.26％ ±0.43％ and 0.00％ ±0.00％,13.44％ ± 0.84％,29.03％ ± 0.78％,57.49％ ± 1.70％,with significant differences (F =174.33,P ＜0.001;F =1 041.40,P ＜0.001),and the following contrast between each two groups met the statistical significance (all P ＜ 0.01).The migration cells per field of CNE2 cells treated with 0,20,40,80 μg/ml osthole for 48 hours were 52.13 ± 4.49,29.00 ± 4.49,18.50 ± 1.93,13.75 ± 2.77,which exhibited a significant difference (F =200.37,P ＜ 0.001),and the following contrast between each two groups met the statistical significance (all P ＜ 0.01).The invasion cells per field of CNE2 cells treated with 0,20,40,80 μg/ml osthole for 48 hours were 46.63 ± 2.87,24.13 ± 2.87,16.75 ± 5.29,11.00 ± 1.77respectively,which exhibited a significant difference (F =131.92,P ＜ 0.001),and the following contrast between each two groups met the statistical significance (all P ＜ 0.01).Meanwhile,the relative mRNA and protein expressions of E-cadherin in 0,20,40 and 80 μg/ml osthole treated-cells (exposure for 48 hours) were 1.00±0.13,2.61±0.03,3.12±0.09,3.60±0.06 (F=20.92,P＜0.001) and0.22±0.03,0.35±0.01,0.60 ± 0.04,0.82 ± 0.03 (F =178.63,P ＜ 0.001) respectively,and the differences were statistically significant,and further pairwise comparison showed the differences were statistically significant (all P ＜ 0.05).Furthermore,the relative mRNA and protein levels of vimentin,β-catenin,cyclin D1 in 0,20,40 and 80 μg/ml osthole treatment for 48 hours were statistically significant difference (mRNA level of vimentin:1.00±0.12, 0.68±0.03 0.56±0.01 0.40±0.09,F=9.48,P＜0.010;mRNA level of β-catenin:1.00±0.14.0.78±0.04, 0.69±0.07 0.46±0.12,F=4.84,P＜0.050;mRNA level ofcyclin D1:1.00±0.09, 0.82±0.03 0.58 ±0.09 0.40±0.03,F=9.49,P＜0.010;protein level ofvimentin:0.85 ± 0.02 0.74 ± 0.01, 0.34 ± 0.01 0.27 ± 0.01,F =610.58,P ＜ 0.001;protein level of β-catenin:0.83 ± 0.00 0.44 ± 0.02, 0.39 ± 0.00 0.23 ± 0.03,F =985.74,P ＜ 0.001;protein level of eyclin D1:0.86 ±0.02, 0.67 ±0.00, 0.35 ±0.01 0.25 ±0.01,F=910.57,P＜0.001),and further pairwise comparison showed the differences were statistically significant (all P ＜ 0.05).Conclusion Osthole can inhibit the proliferation,invasion and migration of CNE2 cells,which is related to the regulation of Wnt/β-catenin signal pathway and then suppressing of EMT.

OBJECTIVETo evaluate the effect of bear bile powder (BBP) on angiogenesis, and investigate the underlying molecular mechanisms.

METHODSA chick embryo chorioallantoic membrane (CAM) assay was used to evaluate the angiogensis in vivo. Human umbilical vein endothelial cells (HUVECs) were treated with 0, 0.25, 0.5, 0.75, and 1.0 mg/mL of BBP for 24, 48 and 72 h, respectively. The 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine the viability of HUVECs. Cell cycle progression of HUVECs was examined by fluorescence-activated cell sorting (FACS) analysis with propidium iodide staining. HUVEC migration was determined by wound healing method. An ECMatrix gel system was used to evaluate the tube formation of HUVECs. The mRNA and protein expression of vascular endothelial growth factor (VEGF)-A in both HUVECs and HepG2 human cells were examined by reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay, respectively.

RESULTSCompared with the untreated group, BBP inhibited angiogenesis in vivo in the CAM model (P< 0.01). In addition, treatment with 0.25-1 mg/mL of BBP for 24, 48, or 72 h respectively reduced cell viability by 14%-27%, 29%-69% and 33%-91%, compared with the untreated control cells (P< 0.01). Additionally, BBP inhibited the proliferation of HUVECs via blocking the cell cycle G to S progression, compared with the S phase of untreated cells 48.05%± 5.00%, 0.25-0.75 mg/mL BBP reduced S phase to 40.38%± 5.30%, 36.54± 4.50% and 32.13± 3.50%, respectively (Pglt; 0.05). Moreover, BBP inhibited the migration and tube formation of HUVECs, compared with the tube length of untreated cells 100%± 12%, 0.25-0.75 mg/mL BBP reduced the tube length to 62%± 9%, 43%± 5% and 17%± 3%, respectively (p< 0.01). Furthermore, BBP treatment down-regulated the mRNA and protein expression levels of VEGF-A in both HepG2 cells and HUVECs.

CONCLUSIONBBP could inhibit the angiogenesis by reducing VEGF-A expression, which may, in part, explain its anti-tumor activity.

Animals ; Bile ; chemistry ; Cell Cycle ; Cell Movement ; Cell Proliferation ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Gene Expression Regulation ; Hep G2 Cells ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Neovascularization, Physiologic ; Powders ; RNA, Messenger ; genetics ; metabolism ; Ursidae ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Objective To establish an animal model of rapid eye movement (REM) sleep deprivation (SD) and an animal model for perifornical nucleus microdialysis and investigate the change of cognition, hypocretinergic system and GABAergic system in rats' hypothalamus after various degrees of REM sleep deprivation and sleep revival and two GABAergic drugs intervention. Methods The modified multiple platform method (MMPM)was used to establish sleep deprivation model and the cognitive function was assessed by Morris' water maze. Immunofluorescence technique was used to analyze the number of Hypocretin (Hcrt) immunoreactive neurons, total Fos immunoreactive neurons, Hcrt and Fos colabeled neurons, and the integrated optical density ( IA ) of GABAA Rαl immunoreactive area in rats' hypothalamus.High performance liquid chromatograph (HPLC) was used to quantitatively analyze the level of GABA and Gluin in the rats' hypothalamus. Two GABAergic drugs, a selective GABAA R antagonist, SR-95531, and a selective blocker of type 1 GABA transporter (uptake blocker), NO-711, were used for perifornical nucleus microdialysis. Results There was no statistically significant difference in tests between CC and TC ( Define CC and TC). There was a significant decrease (P ＜ 0. 05 ) of cognitive function measured by Morris maze test in SD 3 d, SD 5 d and RS 6 h of SD groups compared with CC and TC groups. Number of Fos immunoreactive, F+ &H+ immunoreactive neuronsand IA of GABAA Rαl immunoreactive area were all significantly increased ( P ＜ 0. 05 ). Content of GABA measured by HPLC was also increased ( P ＜ 0. 05 ). However, all these changes were partly reversed by sleep revival SR-95531 and NO-711 had different effect on these changes. Conclusions Sleep deprivation, no matter mild or severe, has adverse effects on cognitive function. Activities of both GABAergic and Hcrtergic system are increased during REMSD. These two neurons system could be regulated by each other and the relationship between them is positive correlation. GABAergic system also had self-regulation during REMSD, but microdialysision of either SR-95531 or NO-711 acquired adverse effects on cognitive function of rats. So GABAergic system is not an optimal therapeutic target. Because GABAergic and Hcrtergic system has inhibitory effect on each other,suppressing activity of Hcrtergic system might be a promising therapeutic target.

Objective To study the effects of sleep inertia (SI) of different time course sleeps on sleep stages and cognitions in healthy men after 30 h sleep deprivation,and also to investigate the mechanism of cognitive functions impairment in sleep inertia stages and the influential factors of sleep inertia.Methods Ten healthy men (age,(20.8 ±2.1) years) participated in the program.The program was divided into 2 stages.First,participants attended a series of tests after 20 min nap(20 min nap group)after 30 h sleep deprivation.The testing series included 3 parts:the continuous performance task,the Stroop Tests,and the Addition Tests.The series of tests were done 3 times immediately after the volunteers were awoken and each lasted about 15 minutes with an interval of 10 minutes between each test.The polysomnogram (PSG) was recorded during the nap.The following 7 days was set as washing-out period to ensure a complete recovery.Participants repeated the similar processes with 2 h nap(2 h nap group) instead of 20 min nap.The cognitive performance of each group was compared with each other along with the best cognitive performance in awakening to estimate whether or not the cognitive abilities regained the normal condition.Results ( 1 ) Sleep latency became shortened in both groups after 30 h sleep deprivation.There were no slow wave sleep (SWS) and rapid eye movement sleep (REM) sleep stages in the 20 min naps,while the percentage of SWS was increased and percentage of REM declined in 2 h naps.(2)In the early of SI (5 min after awaking),cognitive tasks showed that the abilities of continuous attention,selected attention and addition ability were all impaired (continuous performance task:(371.8 ± 21.3 ) times/3 min vs (334.4 ± 22.4) times/3 min,( 373.2 ± 19.0) times/3 min vs ( 323.7 ± 23.8) times/3 min,t =10.443,7.774,both P＜0.01; Stroop tests:(20.3 ±1.5) points vs(17.3 ± 1.0) points,(21.5 ±0.8)points vs( 16.1 ± 1.4 ) points,t =8.478,4.934,both P ＜ 0.05 ; Addition Tests:( 222.2 ± 13.2 ) s vs ( 266.6 ±23.7 ) s,( 226.3 ± 10.9) s vs ( 267.6 ± 23.4 ) s,t =5.748,6.685,both P ＜ 0.01 ).The cognitive functions impairments of 2 h nap group were more severe at the initiation of sleep inertia,but regained the normal condition more quickly.Different cognitive tasks recovered at different speeds.The dispersion of SI needed 30 min.Conclusions ( 1 ) There are difference in the sleep construction and awaked sleep stage between 20 min nap and 2 h nap groups.(2) SI exerts negative influences on cognitive performances of continuous attention,selected attention and addition after sleep deprivation.Many factors may influence the dispersion of SI,including sleep debt,compensation of sleep debt and others.(3) Fragments of sleep are unfavorable to the recovery of body.