This study was aimed to investigate the effects of ursolic acid on human hepatic stellate cells (HSC-LX-2) proliferation and its mechanism.Different doses of ursolic acid were incubated with HSC-LX-2 cellin vitrof or 48 h.MTT was used for the detection of HSC-LX-2 cell proliferation.The expressions of PDGF-ERK signaling pathway associated proteins were measured by western blot.The results showed that the proliferation of HSC- LX-2 cells was inhibited by ursolic acid in a dose-dependent manner.The inhibition rate of 20,30 and 40μmol·L-1 of ursolic acid was 9.1%,42.3% and 62.6%,respectively.The IC50 was 35.2μmol·L-1.After incubated with ursolic acid for 48 h,protein levels of PDGF-R and p-ERK in 30 and 40μmol·L-1 group were significantly decreased when compared with the normal group (P<0.05 orP<0.01),except the ERK protein.It was concluded that ursolic acid can inhibit HSC-LX-2 cell proliferation.Its mechanism may be related to the blockage of PDGF-ERK signaling pathway.

BACKGROUND: Caspase family is viewed as the executive factor of cell apoptosis. Neuronal apoptosis happens probably after spinal cord injury.OBJECTIVE: To observe the changes in caspase-3 expression after spinal cord injury in rats so as to probe into the relationship between it and neuronal apoptosis and provide the evidence on the prop e r time window of intervention on alleviating secondary spinal cord injury.DESIGN: Self-control and mutual-control were designed in animal experiment.SETTING: Department of Traumatic Surgery and Department of Orthopedics of Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: The experiment was performed in Experiment Room of Tongji Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology from January to December 2001, in which, 54SD rats were employed, of either sex, mass weighted varied from 220 to 250 g and provided from Animal Experimental Center of Tongji Medical College of Huazhong University of Science and Technology.rats were divided into the control and injury group. Laminectomy was only done on Ts and T9 in the control and the injury group was subdivided into 9 subgroups, in which, the materials were collected on the 4th and 8th hours and on the 1st, 2rd, 3rd, 7th 14th and 21st days successively, 6 rats in each one. After abdominal anesthesia with 30 g/L pentobarbitol sodium,sternal cord on T8 andT9 segments were exposed with Nystrom method and 50 g weight compressed the front middle region of the spinal cord of such segments with arch smooth metal pad 2.2 mn×5.0 mm for 5 minutes. After injury, artificial bladder urination was done 3 times at 10:00, 16:00and 22:00 successively everyday till the bladder reflex was established.cord was collected at various time spots after spinal cord injury. 4 pieces of spinal cord tissue masses from each group, about 8mm in length, were embedded with paraffin and sectioned continuously. Afterwards, HE staining, immunohistocheistry and TUNEL (TdT-mediated dUTP-biotin nick end labeling) were performed successively. Two rats were sacrificed on ice in each group and central tissue of injured spinal cord was placed in expression was assayed with immunohistochemistry method, neuronal apoptosis was assayed with TUNEL method and linear correlation was used to analyze the correlativity between caspase-3 expression and neuronal apoptosis.pase-3 expression after spinal cord injury in rats of each group.RESULTS: Six rats were maintained in each group and included in result optic microscope: Extensive hemorrhage appeared in 1 hour in injured segment. In 4 to 8 hours, spinal structure began destructive and a large amount of neuronal death appeared. In 24 hours, the destruction of spinal cord became severe and in 7 to 21days, the range of injury was defined with immunohistochemistry in rats of each group: Very few caspase-3 expressions (2.1±0.5) presented in neurons of spinal cord in normal rat. In 8hoursafter spinal cord injury, caspase-3 expression of positive neurons was increased remarkably (89.2±10.5) and up to the peak (189.6±12.7) in 3 days. Caspase-3 expression of positive cell and apoptotic cell appeared alexpression assayed with transcription-polymerase chain reaction (RT-PCR)in rats of each group: Caspese-3 mRNA (0.442±0.024) began increased in 4h, was up to the peak (0.634±0.028) in 48 hours and was restored to be normal (0.351±0.013) in 7 days, which appeared early than apoptosis, indicating positive correlation with the level of neuronal apoptosis (r=0.622).In the control and 4 hours group, stained cell was seen occasionally and positive cell appeared 8 hours later, mainly localized in gray matter. Afterwards, positive cell was increased and up to the peak in 3 days. In 7 days,positive cell of apoptosis and staining was decreased gradually in gray matter, mainly around the white matter. Little amount positive cells appeared on the 14th day and 21st day.CONCLUSION: In normal spinal cord tissue, caspase-3 existed in form of zymogen with very low activity. Caspase-3 is enhanced in expression after spinal cord injury in rats, expresses in large amount in 8 hours and is up to the peak in 24 to 48 hours, which is overlapped in time with positive apoptotic cell assayed with TUNEL and concerning to the localization, it is in conformity with positive apoptotic cell of spinal cord injury compared with positive cell of caspase-3. It is indicated that caspase-3 is involved in regulation of cell apoptosis after spinal cord injury. It is seen in this experiment that the time from spinal cord injury to the activation of caspase-3 is the time window of treatment for cell apoptosis intervened by spinal cord and alleviating secondary spinal cord injury. It is suggested that genetic intervention or specific caspase-3 inhibitor should be applied in 48 hours.

Objective To investigate the mechanism of recombinant human growth hormone(rhGH) in articular cartilage defect repair in vivo.Methods The cylindrical,full-thick articular cartilage defects,(3.5 mm) in diameter,were made in the knee joints of 25 rabbits.rhGH of 0.1 U/kg was injected into the right knee joints 3 times a week for 4 weeks and equal volume of physiological saline to the left knee joints as control.The animals were killed respectively in 4,6,8,12,24 weeks,and the macroscopic,histologic and ultrastractural examinations were performed.Results The repairing process in experimental group was faster and better than that in the control.In the experimental groups,the defect was filled on week 4,the regenerated cartilage mainly in the shallow region and fibrous tissue in the deep region;on week 24,the regenerated tissue was morphologically close to the neighbour normal cartilage,hard to differentiate by macroscopy;Light microscopy showed the defect recovered to normal cartilage structure and the electron microscopy showed a large quantity of mature cartilage cells.In the control groups,the defect was of shallow introcession,mainly of fibrous tissue.The histological examination showed the significant difference between experimental and control groups(P

BACKGROUND:Electrospun polylactic acid/polycaprolactone nanofibers (ENF) are a kind of self-synthesized biodegradable material. Our preliminary studies have indicated that the biomaterial exhibits excel ent biocompatibility;however, the research about its mechanics is stil little. OBJECTIVE:To explore the effects of static pressure on the cytocompatibility of adipose-derived stem cel s on the ENF scaffold. METHODS:Adipose-derived stem cel s were seeded onto the ENF scaffold, and then cultured in the low-glucose DMEM supplemented with 10%fetal bovine serum. The mixed constructs were submitted to the static pressure at 0, 15, 30, and 45 kPa for 4 hours using a static pressure device, respectively. Subsequently, the proliferation, adhesion and viability of adipose-derived stem cel s on the ENF scaffold were detected using MTT assay and living/dead staining to evaluate the cytocompatibility. RESULTS AND CONCLUSION:MTT assay showed that there were significant differences in absorbance values among groups by one-way analysis of variance after 4 hours of loading with different static pressures in vitro. Under 0-30 kPa static pressure, the absorbance values increased with static pressure, but the absorbance values declined until the pressure reached 45 kPa, and multiple comparisons between groups showed significant difference. The significant differences in the cel attachment percentage by MTT assay could be found among groups. The living/dead staining results supported the above findings. Furthermore, the significant differences in percentage of living cel s among groups were shown using either one-way analysis of variance or paired t test. In conclusion, the appropriate static pressure can promote the cytocompatibility, proliferation, adhesion and viability of adipose-derived stem cel s on the ENF scaffold. But the excessive pressure is likely to inhibit the cel ular biological behaviors, thus affecting cytocompatibility of adipose-derived stem cel s with the ENF scaffold.

Objective: To observe the effect of Liuwei Dihuang decoction on brain development of intrauterine growth retardation rats,and to demonstrate the relationship between brain and kidney in TCM.Methods: Animals were divided into 4 groups at random: normal group,model group,Huangqi(HQ) and Liuwei Dihuang(LD) treated groups.The IUGR model was established by passive smoking.On the 19th day of pregnancy,all rats were killed;the total numbers of embryos,the lively,dead and absorbed embryos were counted.The body and brain weight of lively embryos were scaled respectively,then microstructure and apoptosis in brain were observed.Results: Passive smoking can result in the number of dead and absorbed embryos increases.Compared with normal group,the number of apoptotic cells of model group increased.Compared with model group,in Huangqi and Liuwei Dihuang treated groups,the number of dead and absorbed embryos decreased apparently,body and brain weight increased obviously,the number of apoptotic cells reduced significantly(P

This study examined the effects of ω-3 polyunsaturated fatty acid (ω-3PUFA) on the expression of toll-like receptor 2 (TLR2), toll-like receptor 4 (TLR4) and some related inflammatory factors in peripheral blood mononuclear cells (PBMCs) of patients with early-stage severe multiple trauma. Thirty-two patients who were admitted to the Department of Traumatic Surgery, Tongji Hospital (Wuhan, China) between May 2010 and November 2010, and diagnosed as having severe multiple trauma with a injury severity score (ISS) no less than 16, were enrolled in the study and divided into two groups at random (n=16 in each): ω-3PUFA group and control group in which routine parenteral nutrition supplemented with ω-3PUFA or not was administered to the patients in two groups for consecutive 7 days. Peripheral blood from these patients was collected within 2 h of admission (day 0), and 1, 3, 5 and 7 days after the nutritional support. PBMCs were isolated and used for detection of the mRNA and protein expression of TLR2 and TLR4 by using real-time PCR and flow cytometry respectively, the levels of NF-κB by quantum dots-based immunofluorescence assay, the levels of TNF-α, IL-2, IL-6 and COX-2 by ELISA, respectively. The results showed that the mRNA and protein expression of TLR2 and TLR4 in PBMCs was significantly lower in ω-3PUFA group than in control group 5 and 7 days after nutrition support (both P<0.05). The levels of TNF-α, IL-2, IL-6 and COX-2 were found to be substantially decreased in PBMCs in ω-3PUFA group as compared with control group at 5th and 7th day (P<0.05 for all). It was concluded that ω-3PUFA can remarkably decrease the expression of TLR2, TLR4 and some related inflammatory factors in NF-κB signaling pathway in PBMCs of patients with severe multiple trauma, which suggests that ω-3PUFA may suppress the excessive inflammatory response meditated by the TLRs/NF-κB signaling pathway.

BACKGROUND:The repair and management of ful-thickness skin defects resulting from burns and chronic wounds remain a significant unmet clinical chal enge. Using epidermal stem cel s and keratinocyte growth factor for ful-thickness wound repair is a promising approach. Low-frequency electromagnetic fields which are a non-invasive physical stimulation therapy have been recognized as a good method to enhance wound healing. OBJECTIVE:To develop a new strategy to accelerate wound healing by transplanting transfected epidermal stem cel s and keratinocyte growth factor and treating with low-frequency electromagnetic fields in a mouse model. METHODS:Epidermal stem cel s from Sprague-Dawley neonatal rats were isolated and cultured in vitro, then the cel s were labeled with 5-bromo-2-deoxyuridine and transfected by Ad-KGF, a recombinant adenovirus carrying the keratinocyte growth factor. Mice were given to create ful thickness skin wound on the dorsum and randomly assigned to four groups:control group, transplantation of epidermal stem cel s group, transplantation of keratinocyte growth factor gene modified epidermal stem cel s group, and transplantation of keratinocyte growth factor gene modified epidermal stem cel s plus low-frequency electromagnetic field exposure group. RESULTS AND CONCLUSION:The best healing pattern was observed in the keratinocyte growth factor gene modified epidermal stem cel s plus low-frequency electromagnetic field exposure group (P<0.05) at days 9 and 16. 5-Bromo-2-deoxyuridine labeled cel s existed in the wound in the treated groups at day 9. A significantly increased expression of endogenous keratinocyte growth factor was detected in the transplantation of Keratinocyte Growth Factor gene modified epidermal stem cel s group, and transplantation of keratinocyte growth factor gene modified epidermal stem cel s plus low-frequency electromagnetic field exposure group at day 16. A wel-advanced epithelialization was observed in transplantation of keratinocyte growth factor gene modified epidermal stem cel s plus low-frequency electromagnetic field exposure group at days 16 and 30. These results suggest that low-frequency electromagnetic fields enhanced wound healing fol owing the transplantation of keratinocyte growth factor gene modified epidermal stem cel s.

Objective To investigate the efficacy of combined monitoring of motor evoked potentials with transcranial electrical stimulation (TES-MEP),somatosensory evoked potentials (SEP) and spontaneous electromyo-graphy (s-EMG) in tuberculosis surgery involving the thoracic,lumbar and sacral vertebrae.Methods Twenty-seven patients with tuberculosis of the thoracic vertebrae (T2-L2) received intra-operative SEP and TES-MEP monito-ring.Combined SEP,TES-MEP and spontaneous EMG monitoring were employed in 11 patients with tuberculosis of the lumbar or/and sacral vertebrae (L3-S1).SEP and TES-MEP were used to precisely observe the status of the sen-sory and motor pathways; s-EMG responses were used to more accurately localize nerve root irritation.ResuIts (1) SEP monitoring was successful in all of the operations.TES-MEPs were successfully monitored in 35 of them (92.1％).Combined motor and sensory monitoring was successfully achieved in 35 cases (92.1％).Abnormal SEPs were observed in 3 cases (7.9％),while abnormal MEPs were observed in 11 cases (28.9％).Abnormality in both the SEP and TES-MEP occurred in 2 cases (5.3％).There were 9 cases (23.7％) where the SEPs were nor-mal and the TES-MEPs were abnormal.In only 1 case (2.6％) was the SEP normal and the MEP abnormal.The false negative rate was 0％ with combined SEP and TES-MEP monitoring,while the false positive rate was 5.3％.There were 2 cases complicated by post-operative neurological deficits.(2) Spontaneous EMG monitoring can accu-rately determine the functioning of lumbar nerve roots during lumbar or lumbosacral tuberculosis surgery.Among 5 cases where EMG responses were observed,4 cases occurred during the spinal canal and nerve root decompression,1 case occurred in the orthopedic reset phase.Conclusions (1) During tuberculosis surgery involving thoracic,lumbar or sacral vertebrae,combined monitoring of SEPs and TES-MEPs can reflect the physiological and pathological condition of the spinal cord after ruling out interfering factors.This can improve monitoring and help assure the safety of lumbar surgery.(2) Intra-operative s-EMG monitoring can accurately reveal nerve root function in real time,help-ing to avert nerve root injury in lumbar and lumbosacral tuberculosis surgery.

Objective To observe the effect of cognitive rehabilitation training based on games on cognition of the traumatic brain injury (TBI) patients.Methods 60 cases of patients according with the inclusion and exclusion criteria were divided into the control group and the training group with 30 cases in each group.The Montreal cognitive assessment (MoCA) test scores of all patients were lower than 26 points.The training group accepted the cognitive rehabilitation training based on games such as exactly the same game,drum game and picture memory game,while the control group did not accept the training.After one month,all patients accepted the MoCA test again.Then we analyzed the differences of the cognition between the two groups.Results After 30 days of training,all items of the cognitive function increased except the sub-item of abstraction.While in the control group,only the scores of attention,delayed recall,orientation and the total score showed alleviation.And all the scores of the training group were higher than those of the control group except the sub-item of abstraction.Conclusions Cognitive rehabilitation training based on games can effectively improve the cognitive function of TBI patients.

Objective To investigate the incidence, clinical symptoms, correlative risk factors and prognosis of dysautonomia in patients with severe traumatic brain injury. Methods A total of 142patients with severe traumatic brain injury treated from January 2008 to March 2010 were retrospectively surveyed to compare the clinical features of dysautonomia group and control group. Logistic regression was used to analyze the risk factors for dysautonomia. At 6 months post-trauma, the Glasgow Outcome Score (GOS) was used to measure the outcome. Results Of all the patients, 94 patients survived and were followed up. There were 16 patients ( 17% ) diagnosed as dysautonomia depended on clinical symptoms,with statistical difference in aspects of GCS, coma duration, ICU time and average length of stay (ALOS)(P < 0.05). The patients with dysautonomia tended to have poorer outcome ( P < 0.05 ) and showed a positive association with diffuse axonal injury (DAI) ( OR = 11. 25, CI 7.65-16.54 ). Conclusion Dysautonomia has high incidence and is usually severe in patients with severe traumatic brain injury,when DAI may contribute to its occurrence and result in poor prognosis.