Aim To observe the protective effects of epigallocatechin gallate(EGCG)on cortical neurons of embryonic mice the injury by FeSO4/H2O2 in vitro.Methods The cell injury model was established with FeSO4/H2O2 for 24 h.The process of the growth and morphology of the neurons were observed under phase contrast microscope in vitro.The cell survival was analyzed using colormetric neutral red assay.The cortical neuronal viability was analyzed using MTT assay.TBARS method was performed to detect the levels of Malondialdehyde(MDA).The composing capacity of total protein was measured by BCA method.Results As compared with FeSO4/H2O2,EGCG enhanced the mouse cortical neuronal survivality and viability,decreased the levels of MDA and increased the composing capacity of total protein.Conclusion EGCG exerted protective effects on cortical neurons the injury induced by FeSO4/H2O2,which may be related to free radial scavenging activity and lipid peroxidation process being inhabited.

Objective To discuss the repairing methods of the wound after upper lip lesion excision.Methods The wound after upper lip lesion excision was repaired by expanded pedicled submental flap.The 3 cm-long incision was located in 1 cm to sub-mandible.The 100 ml expander was placed beneath the platysma,and the aqueducts and spigots of the expanders were laid out of the skin.After complete expansion,the spastic scars of the upper lip and nasal bottom were resolved,the nasal columella and upper lip were put back to the normal position.The pedicled submental flap was transferred to the wound after upperlip excision according to the size of the wound.The pedicle was severed after 3 weeks.Results There were 5 cases of the expanded pedicled submental flap to repair the wound after upper lip excision.The flap survived without complications.The appearances were satisfied by the patients.Conclusions The method of the expanded submental flap is suitable for the wound after upper lip excision.

BACKGROUND:To improve local microenvironment and reduce local scars is conducive to peripheral nerve regeneration that promotes nerve function recovery.OBJECTIVE:To evaluate the effect of fresh amniotic membrane on the regeneration of tinjured peripheral nerve.METHODS:Sixty healthy Sprague-Dawley rats were randomly divided into three groups (n=20 per group) after constructing a model of sciatic nerve injury of the unilateral leg. In group A, the nerve was wrapped with fresh human amnion at the anastomosis end after the repair of nerve. In group B, the nerve was wrapped with biofilm at the anastomosis end after the repair of nerve. In group C, no treatment was conducted after the repair of nerve (blank control). The effects were evaluated by anatomical observation, light microscope observation, immunohistochemical detection (2, 4, 8, 12 weeks after surgery), transmission electron microscope observation, axon imaging analysis, action potential detection, and sciatic nerve function index (4, 8, 12 weeks after surgery).RESULTS AND CONCLUSION: (1) Gross observation. The amniotic membrane and biofilm were absorbed partialy at postoperative 2 weeks, mostly at postoperative 4 weeks and completely at postoperative 8 weeks. In the groups A and B, the nerve was adhered slightly and loosely to the surrounding tissues, with a fair range of motion. In the group C, the nerve was tightly adhered to the surrounding tissues, with a poor range of motion. (2) Observation under light microscope. The nerve regeneration was better in the groups A and B than group C at 2, 4, 8, 12 weeks postoperatively. (3) Observation under electron microscope. Regenerated nerve fibers were rarely seen and lamelar structures were unclear in the three groups at 4 weeks postoperatively. Then, increased regenerated nerve fibers, thickened myelin sheath, clear lamelar structure and enlarged axon diameter were found in the groups A and B compared with the group C at 8 and 12 weeks postoperatively. (4) Immunohistochemical detection. The expression and distribution of S-100 protein in the groups A and B were better than those in the group C. (5) Axon image analysis. Groups A and B were superior to the group C in the diameter of myelinated nerve fibers, thickness of myelin sheath and number of regenerated nerve fibers. There was a significant difference by statistical analysis (P < 0.05). (6) Electrophysiological examination. Shorter latency period, higher amplitude and faster nerve conduction velocities were observed in the groups A and B compared with the group C (P < 0.05). (7) The sciatic function index. The sciatic function index in group A or B was significantly higher than that in group C (P < 0.05). To conclude, the human amniotic membrane can reduce adhesion between the damaged nerve and surrounding tissues, and prevent scarring at the anastomosis end. In addition, it promotes the regeneration of nerve fibers, increase axon diameter and myelin sheath thickness, and ease inflammatory and immune responses at the neural incision.

BACKGROUND:Experiments have demonstrated that biological membranes can be usedtorecon struct thetendon she athandin hibit exogenou shealing of thetendon.Therefore,the semembrane sprovide a good bed for tendon gliding and reduce tendon adhesion.
OBJECTIVE:To compare the effectsof acelular amniotic membrane and medical membraneagainst tendon adhesion during the repair oftendon sheath defects.
METHODS:ToesIIIfrom the bipeds of 66 leghorns were chosen to prepare tendon injury and tendon sheath defect models, which were randomly divided into three groups (n=22 per group). Amnion group were repaired with acelular amniotic membrane, medical membrane group with absorbable membrane, and control group had no treatment on tendon sheath defects. Gross, histological and biomechanical tests of each group were performed at 2, 4, 8, 12 weeks after surgery.
RESULTS AND CONCLUSION:At 12 weeks after surgery, in the amniotic membrane and medical membrane groups, the tendon sheath formed completely, and the tendon healed well, with no adhesion, but in the control group, there was serious tendon adhesion. At 8 weeks after surgery, the number of synovial cells in the false sheath was highest in the amniotic membrane group sequentially followed by the medical membrane group and control group. In the amniotic membrane group, the rough endoplasmic reticulum expanded highly and secreted exuberantly in the matrix, while in the control group, the synovial cells presented with messy arrangement, and expanded vacuoles in the matrix were weaker than those in the other two groups. At 12 weeks after surgery, fibroblasts were arrayedtidily in layerwith dense structure in the medical membrane and amniotic membrane groups;but in the control group, fibroblasts were distributed disorderly with loose structure. Tendon sliding distance and total flexor toe angle in the amniotic membrane and medical filmgroups were significantly larger than those in the control group (P < 0.05),butthere was no significant difference between the medical membrane and amniotic membrane groups. Additionally, the maximum tensile fracture strength had no significant difference among three groups at 12 weeks after surgery. These results indicate that both amniotic membrane and medical membrane can markedlyprotect the tendon from exogenous healing and adhesion.

Objective To analyze the expression feature and function of microRNAs in exosomes secreted by leukemia cells (LCEX). Methods The mice leukemia cell line L1210 was taken as the example, and LCEXL1210 was obtained by isolating supernate of L1210 cells through density gradient centrifugation. MicroRNAs isolated from LCEXL1210 were analyzed by microarray analysis, compared with miRNA from L1210 cell line, and then some of miRNAs with different expression were verified by real-time PCR and were analyzed by Gene Ontology (GO) database. Results The number of miRNAs identified in LCEXL1210 was 1 044, and that in L1210 cell line was 872. The number of shared miRNAs between LCEXL1210 and L1210 cell line was 732, accounting for 70.1 % of LCEXL1210 and 83.9 % of L1210 cell line, respectively, which indicated that 70 % of LCEXL1210 was derived from the parental cells. Interestingly, 312 miRNAs in LCEXL1210 were found to be underrepresented in the parental cells, indicating their specificity in LCEXL1210. Some miRNAs were significantly highly expressed in LCEXL1210 compared with those in L1210 cell line, including miR-16-1, miR-210, miR-195 and so on, which showed that miRNAs isolated from LCEXL1210 were differentially expressed with those from the parental cells. Some differentially expressed miRNAs from LCEXL1210 were verified by real-time PCR, and then were analyzed by GO database, which demonstrated that these highly expressed miRNAs participated in the processes of various biological function and signal transduction. Conclusions MiRNAs isolated from LCEXL1210 show a high similarity to miRNAs isolated from L1210 cells, whereas of which one-third are specific. The highly expressed miRNAs participate in the processes of various biological function and signal transduction.

Objective To explore the repair method for wounds after excision of facial benign tumors.Methods Unilateral or bilateral deltopectoral skin flaps were expanded depending on the area of the facial benign tumor.Expander was implanted underneath deltopectoral flap region through an incision inferior to the clavicle.When expansion was completed,all or part of the benign tumor was excised before designing the flap according to the area of the skin defect.The area of the skin flap should be more than that of skin defect with 10％ to 15％.The pedicle wound could be sealed by rolling it around to form a tube or a hinge using the benign tumor and pedicle.The flap was delayed three weeks later and the pedicle was divided one week after flap delaying.Results All 20 cases got the satisfactory results with treatment of pedicled expanded deltopectoral skin flaps for repair of wounds after excision of facial benign tumor.Conclusions It is a better option to repair a large area wound after excision of facial benign tumor with an expanded deltopectoral skin flap.

Objective To investigate the application of the frontal branch of superficial temporal vessels pedicled flap in repairing lower eyelid ectropion.Methods Eight cases were collected from patients diagnosed with lower eyelid ectropion in our hospital from April 2012 to April 2015.In phase 1 of operation,the dilators were implanted into the frontal branch of superficial temporal vessels and fully expanded by normal saline injection;In phase 2,the scar of lower eyelid was incised,and the expanded forehead flaps were transferred to cover the wound after the lower eyelid released back to normal anatomy location;In phase 3,the flap delay operation was manipulated 3 weeks after phase 2,and the left wound after scar excision was finished by pedicle division 1 week later.Results All patients in the study showed a good appearance and function of lower eyelid.There were no complications such as flap congestion and necrosis occurred.Meantime there were no relapses observed according to the follow-ups ranging from 6 months to 1 year.Conclusions The application of the frontal branch of superficial temporal vessels pedicled flap shows a promising procedure in treatment of lower eyelid ectropion.

Objective To analyze the genome sequencing of mumps virus strain 79 purified from working seeds plague and examine its phylogenetic relationship with wild type virus strain isolated in China,and to explore the efficacy of vaccine at molecular level.Methods Whole genome sequences of two substrains of S79(major and minor),were obtained with fragment amplification by RT-PCR.Genetic distances between S79 and strains identified in China were analyzed based on the phylogenic analysis of small hydrophobic protein(SH) sequences.Results The nucleotide homologies of two S79 substrains(major and minor),with a ratio of 2:5 during culture,with Jeryl Lynn(JL) strain were 99.7% and 100%,respectively.There were scattered non-homologous recombination between two substrains.The genetic distances between strain S79,which was genotype A,and wild type virus strain identified in China,which were genotype F,was 11.2% -20.0%.S79 live vaccine was composed of two substrains,major and minor component,which were highly similar to JL strain in their genome sequences,but different from JL in their ratios during culture.Conclusion Different from wild type virus strain identified in China,the genotype of S79 was A,and phylogenetically distant from other strains,which may account for the low efficacy of S79 live vaccine.The ratios of two substrains might also be of interest for further study of the vaccine protection and efficacy.

Objective To explore the expression of miRNA-181a (miR-181a) in patients with multiple myeloma (MM) and its effect on biological features of MM cells. Methods CD138+cells of bone marrow from 25 MM patients and 10 patients with hematological non-malignancies were purified by using immunomagnetic separation, and the expression of miR-181a in CD138+cells and MM cell lines including RPMI 8226, H929 and U266 were detected by real-time quantitative PCR. The effects of down-regulation and up-regulation of miR-181a expression on the biological characteristics of MM cells were studied with miR-181a antagomir and agomir. Results Compared with patients with hematological non-malignant diseases, the expression of miR-181a in CD138+ cells was upregulated in MM patients. Compared with CD138+ cells in hematological non-malignancies, high expressions of miR-181a were observed in RPMI 8226 and U266 myeloma cell line, while low expressions of miR-181a were observed in H929 cells. Down-regulation of miR-181a with 100 nmol/L miR-181a antagomir could inhibit the proliferation of U266 cells at 24,48 and 72 h [(67.1 ± 3.3) %vs. (50.5 ± 4.1) %, (71.5 ± 3.6) % vs. (52.3 ± 2.2) %, (78.1 ± 5.4) % vs. (69.5 ± 4.3) %, P < 0.05 respectively], whereas up-regulation of miR-181a with 100 nmol/L miR-181a agomir could significantly promote the proliferation of H929 cells at 24 h and 48 h [(21.2 ± 2.4) %vs. (38.5 ± 3.6) %, ( 61.3 ± 5.4) %vs. (82.2 ±6.9)%, P<0.01 respectively]. Cell cycle analysis showed that miR-181a antagomir made U266 cell cycle arrest in the G0/G1 phase. Meanwhile, susceptibility test results indicated that the apoptosis of U266 cells induced by doxorubicin, paclitaxel and 5-fluorouracil was increased when the proliferation of miR-181a expression was down-regulated with miR-181a antagomir. In migration assay, the data showed that down-regulation of miR-181a with miR-181a antagomir could inhibit the migration of U266 cells, and the proportion of migrated cells in the experimental group (62 ± 10) %was lower than that in the control group (89 ± 12) %(P< 0.05), whereas up-regulation of miR-181a with miR-181a agomir could improve the migration of H929 cells, and the proportion of migrated cells in the experimental group (242 ± 9) % was higher than that in the control group (98 ± 8)%(P<0.01). Conclusions The high expression of miR-181a expressed highly by MM cells may promote the proliferation, migration and drug resistance of myeloma cells, indicating that miR-181a could be an important prognostic biomarker candidate, and the application of gene silencing may improve the prognosis of MM.

OBJECTIVETo investigate the anti-oxidative stress and anti-fibrotic mechanisms of adiponectin by examining effects on oxidative stress levels and expression of fibrosis-related signaling factors, including transforming growth factor-beta 1 (TGFb1), collagen I (COL-1), and the adenosine monophosphate-activated protein kinase (AMPK) pathway by using an in vitro HSC-T6 cultured cell system.

METHODSActivated HSC-T6 cells were pre-treated with 1.0 mug/mL adiponectin for 0, 30, 60 and 120 min, or left untreated to serve as controls, and both groups were then exposed to 5 mumol/L H2O2; a portion of the adiponectin-treated oxidative stress-induced cells were treated with an AMPK inhibitor (Compound C). The effects on mRNA levels of TGFb1. and COL-1 were analyzed by real-time PCR, in the levels of secreted TGF-b1 and COL-1 were detected by enzyme-linked immunosorbent assay of supernatants, and in the phosphoAMPK and AMPK protein expressions were detected by Western blotting.

RESULTSCompared to the H2O2 group without adiponectin pre-treatment, the H2O2 group with adiponectin pre-treatment showed significantly increased activity of superoxide dismutase (SOD), decreased content of malondialdehyde (MDA), and decreased gene and protein expressions of TGF-b1 and COL-1 (P less than 0.05). Moreover, inhibition of the AMPK pathway inhibited these adiponectin-mediated effects. The H2O2 group with adiponectin pre-treatment also showed increased levels of phospho-AMPK protein expression, with the maximum effect detected after 120 min of the adiponectin pre-treatment (P less than 0.01).

CONCLUSIONInhibition of oxidative stress is one of the mechanisms of the anti-fibrotic effects of adiponectin. Adiponectin can attenuate oxidative stress levels, resulting in down-regulation of TGFb1 and COL-1 expression through activation of the AMPK pathway.

AMP-Activated Protein Kinases ; Adiponectin ; Cell Line ; Collagen Type I ; Down-Regulation ; Hepatic Stellate Cells ; Humans ; Hydrogen Peroxide ; Oxidative Stress ; RNA, Messenger ; Signal Transduction ; Transforming Growth Factor beta1