Objective To investigate the effects of extractum trametes robiniphila murr combinated with hydroxyurea on proliferation and apoptosis of K562 cells and whether the 2 drugs have synergistic effects,and to detect the antiproliferation and apoptosis ratio of cells in diffe-rent groups were detected.Methods K562 cells were cultured in vitro,and logarithmic phase cells were used for study.MTT-inhibitory test and cell morphological analysis were employed to examine the effects of extractum trametes robiniphila murr alone or in combination with hydroxyurea on proliferation of K562 cells.The effects of extractum trametes robiniphila murr alone or in combinated with hydroxyurea on apoptosis of K562 cells were examined by flow cytometry.The expressions of bcr-abl,bax and bcl-xl mRNA levels were detected by way of reverse transcription polymerase chain reaction(RT-PCR).Results Extractum trametes robiniphila murr could inhibit proliferation and induce apoptosis of K562 cells,and extractum trametes robiniphila murr combinated with hydroxyurea had a synergistic effects on cell prolife-ration and apoptosis.The RT-PCR displayed that either extractum trametes robiniphila murr alone or in combinated with hydroxyurea could down-regulate the expression of bcl-xl mRNA and up-regulate the expression of bax mRNA and the effect of the 2 drugs had a synergistic effect on bcl-xl and bax mRNA expressions.Extractum trametes robiniphila murr alone could also down-regulate the expression of bcr-abl mRNA,but hydroxyurea alone did not show any effect on the expression of bcr-abl mRNA and the 2 drugs had no synergistic effect.Conclusions The effect of extractum trametes robiniphila murr combinated with hydroxyurea on the proliferation and apoptosis of K562 cells has a synergistic effect.The mechanism of antiproliferation and inducing apoptosis of extractum trametes robiniphila murr is probably related to down-regulating the expression of bcl-xl and bcr-abl mRNA and up-regulating the expression of bax mRNA.The synergistic effect of antiproliferation and inducing apoptosis of extractum trametes robiniphila murr combinated with hydroxyurea on K562 cells is probably related to down-regulating the expression of bcl-xl mRNA and up-regulating the expression of bax mRNA.

[Objective]On May 12,2008,an over 8.0-magnitude earthquake occurred in Wenchuan county,Sichuan province,China.The anthor's field hospital was deployed to An county,which arrived on the 3rd day after the disaster.During the first week of the deployment,284 victims with musculoskeletal injuries received orthopedic management.The objective of this article is to highlight the experience gained by this author's medical assistance team,focusing on the orthopedic care in the early part of the post-disaster relief activities.[Method]Records of 284 patients who underwent orthopedic management were reviewed retrospectively.[Result]There were a large number of musculoskeletal injuries amongst the survivors in the acute phase of the disaster,including spinal trauma,fractures and dislocations,peripheral nerve injuries and soft tissue injuries.The average age was 49.3 years(range,11 to 92 years) with 133 men and 151 women.The patients with upper extremity fracture were 85,with lower extremity fracture 73,with spinal fracture 23,with shoulder or hip dislocation 7.The patients with hand or foot injuries,peripheral nerve injuries and soft tissue injuries were 96.Open injuries were 67,the remaining were closed injuries.Totally 131 patients recovered,and the rest were transferred to the rear hospital for further management.The X-ray postoperatively showed that 21 open fractures were reduced functionally by means of external fixator,and the shoulder or hip dislocations were reduced anatomically.The wounds with primary suture had no infection.There were no other complications.[Conclusion]Correct diagnsis of the injuries,providing scientific and resonable therapeutic measures,and correct operative indication should be emphasized for early orllopedic care.Definitive surgeries such as open reduction and internal fixation are not recommended strongly.External fixation for post-earthquake open fractures is simple and effective,allowing simple approach to wound care and transferring to rear hospital during the whole period of fixation.

Objective To investigate the effect of propofol post?conditioning on oxygen?glucose deprivation and restoration ( OGD∕R)?induced abnormal cell cycle activation in hippocampal neurons of rats. Methods Primarily cultured hippocampal neurons obtained from fetal Wistar rats were cultured for 7 days and seeded in culture wells (100 μl∕well) or in culture flasks (3 ml∕flask) at a density of 5×105cells∕ml. The neurons were divided into 3 groups ( n=24 each) using a random number table: control group ( group C); group OGD∕R; propofol post?conditioning group (group PP). The neurons were subjected to oxygen?glucose deprivation for 1 h followed by restoration of oxygen?gulcose supply for 24 h. Propofol 1.2μg∕ml was added immediately after onset of oxygen?glucose restoration, and the neurons were incubated for 2 h in group PP. At 24 h of oxygen?glucose restoration, cells were collected for measurement of the cell viability by methyl thiazolyl tetrazolium assay, and the mitochondrial membrane potential ( MMP ) , intracellular Ca2+concentration ( [ Ca2+] i ) and distribution of cell cycle were determined using flow cytometry. Results Compared with group C, the cell viability and MMP were significantly decreased, [ Ca2+] i was signifi? cantly increased, the proportion of the cells in G0∕G1 phase was significantly decreased, and the proportion of the cells in S and G2∕M phases was significantly increased in OGD∕R and PP groups (P<0.05). Com?pared with group OGD∕R, the cell viability and MMP were significantly increased, [ Ca2+] i was significant?ly decreased, the proportion of the cells in G0∕G1 phase was significantly increased, and the proportion of the cells in S and G2∕M phases was significantly decreased in group PP (P<0.05). Conclusion The mechanism by which propofol post?conditioning reduces OGD∕R?induced apoptosis in hippocampal neurons of rats is associated with inhibition of abnormal cell cycle activation.

Objective To investigate the reliability and effect of using the chimeric flap retrieved by laparoscopic surgery to cover the large defect of the extremities.Methods The debridement and vacuum sealing drainage (VSD) were performed on 18 patients, who were admitted due to the defects of the extremities.The free chimeric peritoneal-deep inferior epigastric artery perforator (DIEP) flap assisted by laparoscopic surgery was transplanted to cover the defect with exposed tendons and/or skeleton.Results The remaining defects of the extremities were 12 cm × 8 cm-30 cm × 17 cm.The peritoneal component of the chimeric flaps measuring 8 cm × 6 cm -14 cm × 10 cm retrieved by laparoscopic surgery was used to cover the tendons, bones and joints.The deep inferior epigastric artery perforator (DIEP) flaps measuring 13 cm × 10 cm-32 cm × 18 cm allowed the cutaneous coverage of wounds.The chimeric flaps survived completely excepting two patients.The two patients experienced partial necrosis of the chimeric flaps and received skin grafting to achieve the wound closure.The function of the injured extremities recovered partially after 6-18 months of follow-up.No abdominal pain, distension, herniation,bulging and intestinal obstruction were recorded.Conclusion The chimeric flap assisted by laparoscopy is a helpful, safe and effective method for reconstruction of large wounds in extremities with exposed tendons and bones.

Objective To observe the change rules of expressions of IL-1β, IL-4, IFN-γ and TGF-β1 in serum and colon tissue in rats with ulcerative colitis of spleen-kidney yang deficiency type; To explore the action in the progress of ulcerative colitis of spleen-kidney yang deficiency type.MethodsComposite method was used to establish rat models with ulcerative colitis of spleen-kidney yang deficiency type. 75 Wistar rats were randomly divided into blank group, model 3 d group, model 7 d group and model 21 d group. ELISA was used to test the levels of IL-1β, IL-4, IFN-γ and TGF-β1 in serum and colon tissue.Results Compared with the blank group, contents of IL-1β and IFN-γ in model group increased significantly (P<0.05,P<0.01), and the contents of IL-4 and TGF-β1 content decreased (P<0.05,P<0.01),especially in the model 7 d group (P<0.01).ConclusionThe pro-inflammatory factor IL-1β, IFN-γ and anti-inflammatory factor IL-4 and TGF-β1 play an important role in rats with ulcerative colitis of spleen-kidney yang deficiency type.

Objective To observe the effects of Sishen Pills on gene and protein expressions of toll-like receptor 2 (TLR-2) and toll-like receptor 4 (TLR-4) of colonic tissue in spleen-kidney yang deficiency rats with ulcerative colitis (UC); To discuss its mechanism of action for spleen-kidney yang deficiency UC. Methods Lavage of senna +intraperitoneal injection of hydrocortisone injection + enema of 2,4,6-trinitrobenzene sulfonic acid/ethanol were used to establish the model of UC of spleen-kidney yang deficiency type. 90 Wister rats were randomly divided into blank group, model group, Salazosulfadimidine group and the Sishen Pills high-, medium- and low-group. Each medication group was intervened by relevant medicine. RT-qPCR was used to detect the expressions of TLR-2 and TLR-4, and Western blot was used to detect the protein expressions of TLR-2 and TLR-4. SOD and MDA in serum were detected. Results Compared with the blank group, SOD activity decreased and MDA content increased in the model group (P<0.01); gene and protein expressions of TLR-2 and TLR-4 increased significantly (P<0.05). Compared with model group, SOD increased and MDA content decreased in all the medication groups (P<0.05, P<0.01); gene and protein expressions of TLR-2 and TLR-4 decreased (P<0.05, P<0.01). Conclusion Sishen Pills can achieve the treatment of UC by improving the metabolism of oxygen radicals in colonic tissue and regulating the immune system of the intestinal epithelium.

Objective To investigate the effects of lymph from ischemic/reperfused intestine on the inflammatory factors and Toll-like receptor 4 (TLR4) ligand high mobility group box-1 (HMGB1) in TLR4 deficient (TLR4-/-) mice.Methods A total of 20 SD rats weighing (300 ±20) g were randomly assigned into two groups:lymph drainage group (group N,lymph drainage for 180 minutes without other treatment) and intestinal ischemia/reperfusion group (group I/R,draining the lymph for 180 minutes while clipping the superiormesenteric artery for 60 minutes followed by 120-minute reperfusion).Thirty-two TLR4-/-mice and thirty-two C57BL/6 wild type (WT) mice were each divided into 4 sub-groups (n =8),injected with different fluids through the caudal vein:group N with normal lymph;group I/R with I/R lymph;group Edt with endotoxin;group HMGB1 with HMGB1 protein.The mice were sacrificed 180 minutes after the injection for sample collection.Results The levels of endotoxin and HMGB1 in the lymph drainage of the group I/R rats were significantly higher than that of the group N rats [(0.034 ± 0.050) Eu/ml vs.(0.017 ± 0.023) Eu/ml,P =0.033;(4.293 ± 0.883) ng/ml vs.(0.509 ± 0.128) ng/ml,P =0.006].In the mice injected with HMGB1,the mucosa thickness and villus height in the ileum of the WT mice were significantly lower than that of the TLR4-/-mice [(335.8±43.2) μmvs.(602.1±37.5) μm,P=0.000;(273.0±31.7) μm vs.(404.5 ± 18.6) μm,P =0.000];in both WT and TLR4-/-mice injected with the I/R lymph drainage,the mucosa thickness and virus height were decreased,but the decrements were significantly lower in TLR4-/-mice;there were no statistically significant differences in the levels of interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),endotoxin,and HMGB1 between the TLR4-/-and the WT mice injected with normal lymph or endotoxin.In the mice injected with I/R lymph drainage,the levels of inflammatory factors in the TLR4-/-mice were significantly lower than those in the WT mice [TNF-α:(28.637 ±5.166) pg/ml vs.(41.917 ±8.175) pg/ml,P=0.000;IL-6:(60.900 ±24.729) pg/ml vs.(110.265 ±28.545) pg/ml,P =0.000].In the mice injected with HMGB1,the levels of inflammatory factors in the TLR4-/-mice were significantly decreased compared with those in the WT mice [TNF-α:(20.865 ± 6.464) pg/ml vs.(31.059 ± 6.204) pg/ml,P=0.004;IL-6:(36.268 ±8.977) pg/ml vs.(76.677 ± 14.099) pg/ml,P=0.000].Conclusions The concentrations of endotoxin and HMGB1 are significantly increased during intestinal I/R in rats.After injection of I/R lymph drainage,endotoxin,and HMGB1,the levels of inflammatory factors and HMGB1 in the mice injected with I/R lymph drainage are significantly higher than those in the mice injected with normal lymph;the levels of inflammatory factors and local damage of intestinal mucosa are significantly reduced in the TLR4-/-mice than in the WT mice.The gut-lymph pathway may play a key role in the intestinal I/R injury.

By participating in the research practice of basic medicine,the innovative consciousness,the scientific thinking and researching ability of seven-year program medical students can be improved. Meanwhile,the generalization of Open Experimental Teaching and further development of self-design experiment also improve the innovation and research ability of medical students,which can serve as a new approach to the quality education of higher medical talents.

Objective To investigate the relationship between CIDE-B change and apoptosis by detecting expression of CIDE-B gene and protein under various injuring conditions(firearm injury and in- cision injury)and comparing neuronal apoptosis.Methods A total of 108 New Zealand rabbits were randomized into firearm injury group,incision injury group and control group.Samples were harvested at different time points after injury for measuring CIDE-B mRNA expression level by RT-PCR,demonstrating morphological distribution of CIDE-B mRNA by in situ hybridization and determining CIDE-B protein level by Western blot.Results Following nerve injury,CIDE-B mRNA expression in spinal cord was en- hanced markedly in the firearm injury group,increased from at day 1,reached peak at day 7,lasted for two weeks and declined after four weeks.CIDE-B mRNA expression was increased late,with small range in the incision injury group.The changes in CIDE-B protein level accorded with those in CIDE-B mRNA level.There was slight CIDE-B mRNA expression,distributing in spinal cord neuronal cytoplasma,in spinal cord tissues in the control group.Conclusions Following peripheral nerve injury,expressions of CIDE-B mRNA and protein are enhanced in spinal cord neurons and distributed in the grey matter of spi- nal cord.Such enhancement is more marked after firearm injury than incision injury.

Objective:To identify metastasis-associated genes in salivary gland adenoid cystic carcinoma(ACC).Methods:Salivary gland adenoid cytic carcinoma cell line ACC-2 and its highly metastatic ACC-M cells were used to screen the metastasis-related genes in ACC by microarray technology.Two fluorescent cDNA probes labeled with Cy3 and Cy5 dyes,were prepared from the mRNA samples of ACC-2 and ACC-M cells by reverse transcription method.The two color probes were then mixed and hybridized on the cDNA chip constructed by double dots of 1152 human genes,and scanned at two wave lengths.Differentialy expressed genes of the two cell lines were analyzed using computer.Then seven of the differently expressed genes were further validated by RT-PCR technique.Results:Of the 1,152 known genes and expressed sequence tags,26 showed significantly different expression level(minimum 2 fold) between the two cell lines.Among the 26 genes,19 were up regulated(with ratio more than 2) and 7 were down(with ratio less than 1/2).The results of RT-PCR analysis for 7 differently expressed genes were coincident with those of microarray assay.Conclusions:Down regulation of LIFR,LCP1,DPEP1 and ABLIM1,and up regulation of DCC,MMP1 and CNTN2 may be related to the highly metastatic potential of ACC-M cell line.