BACKGROUND:. Growth factor starts a new epoch of the research and application of wound healing, and the effect of the combination of Chinese tradition medicine and biomedicine in wound healing is still in the discussion.OBJECTIVE: To observe the effect in promoting wound healing of rats with bletilla colloid, a Chinese herbal medicine, as a carrier of exogenous high molecular weight nerve growth factor (HMW-NGF).DESIGN: The single sample observation was adopted in the identification of activities of HMW-NGF; The randomized control experiment with animal as subjects was adopted in the observation of the effect of the HMW-NGF mixed with the bletilla colloid as a carrier in promoting wound healing.SETTING: Department of Orthopaedics, Shenzhen Buji People's Hospital,and Department of Orthopaedics, Tongji Hospital Affiliated to Huazhong University of Science and Technology.MATERIALS: Forty SD rats aged 90-120 days of either gender were employed, with the body mass of 220-280 g; The freeze dried preparation of cattle seminal fluid with HMW-NGF with 1mg each.METHODS: From September 2001 to December 2004, the experiment was carried out in the Department of Orthopaedics in Shenzhen Buji People's Hospital and Central Laboratory of Department of Orthopaedics in Tongji Hospital Affiliated to Huazhong University of Science and Technolcolloid, HMW-NGF, HMW-NGF+ bletilla colloid were added into the serum-free medium (SFM), and the influences of them on the dorsal root lected and divided into 4 groups with 10 rats in each group. The rats of normal control group had no medicine on the wound. In the bletilla colloid group, the HMW-NGF group and the HMW-NGF+bletilla colloid group, incision size 2 cm was made at the back of each rat, and bletilla colloid,HMW-NGF, HMW-NGF+ bletilla colloid were applied respectively on the wound, once per day. The wound area was measured at day 3 and 10 after treatment, and the wound area was calculated by the image analysator. The category and the quantity of the emigrated cells were observed with light microscope, and the healing time of the wound was observed.tion under light microscope.the DRG of chick embryo was cultured, the reaction of evection growth around the ganglion in the HMW-NGF, HMW-NGF+ bletilla colloid with the dilution desity of 1:106-1: 108 was most significant, which was not seen in healing with HMW-NGF + bletilla colloid was significantly increased as compared with that in normal control group, the bletilla colloid group and the The healing time of the wound of the normal control group, the bletilla colloid group and the HMW-NGF group and the HMW-NGF+ bletilla colloid group were (19.5±0.7), (17.3±0.6), ( 16.6±0.7 )and ( 14.9±0.4 ) days respecvation of cut sheets 3 days after treatment: The quantity of polymorphonuclear leucocytes, histoleueocyteand fibroblast in the bletilla colloid group,the HMW-NGF group and the HMW-NGF+ bletilla colloid group were significantly larger than that in the normal control group, and the micrangium could be seen in the HMW-NGF group and the HMW-NGF+ bletilla colloid group. The observation of cut sheets 10 days after treatment: The granulation tissue in the HMW-NGF group and the HMW-NGF+ bletilla colloid group was compacter and had more micrangium as compared with the normal control group and the bletilla colloid group.CONCLUSION: HMW-NGF+ bletilla colloid has significant effect on promoting the healing of the wound at the earlier and later period, and the effect is better than that of bletilla colloid or HMW-NGF.

Objective We established a model of human breast cancer in nude mice, to discuss the feature of pathology and biology of breast cancer,and give help to establish tools of pathogen research. Methods Human breast cancer cells MCF-7 were subcutaneously injected into the right armpit of nude mice to establish human breast cancer models.The mice were divided into composite group, estrogen group, leptin group and the blank group (30 in each). In the composite group,estrogen and leptin were injected into peripheral region of the tumor daily.In the estrogen group,estrogen was injected.In the leptin group, leptin was injected.In the blank group, physiological saline was injected.Tumor growth was observed, and the volume of the tumor was recorded.The tumor tissues obtained from the mice were pathologically examined. Results (1)The tumor-taking rate of leptin group and the blank group were 46.7% (14/30)and33.3% (10/30) ,and the mode is failure.In composite group and estrogen group they were 96.7% (29/30) and 70% (28/30).There was not significant difference between them ( P < 0.05).(2) The differences of average diameter and volume were statistically significant between the composite group and the estrogen group (P < 0.05).(3) Pathology diagnosis was invasive ductal carcinoma. Conclusions (1) Establishing human breast cancer model in nude mice need the stimulation of estrogen. The tumor-taking rate of nude mice has no relationship with leptin.(2) In the study in vivo, leptin as same as estrogen has stimulating effect on MCF-7 cells proliferation.(3) The transplanted cancer cell partly have the pathology and biology features of the human breast cancer cells and give help to establish tools of pathogen research.

[Objective]To measure adult normal femur specimen in order to obtain accurate femoral resection and match the distal femoral in total knee arthroplasty.[Method](1)Forty-seven adult normal femur specimens of central Chinese were used for taking digital pictures in both anteroposterior and medialateral position.Five angles were measured through drawing lines of fixed point.(2)The exact position of the entry point was defined by the two wires on the specimens.[Result](1)The five angles reflect distal femoral anatomy features on the frontal plane and sagittal plane.(2)The mean value of distance from the entry point to the center of the intercondylar notch was 6.21 mm and the distance from the entry point to the anterior margin of the femoral posterior cruciate ligament(PCL) insertion is found to be 6.7 mm.[Conclusion](1)The distal one-third of femora was found to be outward bowing on frontal plane and forward bowing on Sagittal plane.Especially,the forward bowing is more remarkable than Outward bowing.So,when the position of intramedullary guide stick is decided by distal femoral anatomy,the axis alignments on frontal and Sagittal planes are both important.(2)During total knee afthroplasty the exact position of entry point may be defined by the Center of the intefcondylar notch and femoral PCL insertion.

Objective To evaluate the different effects of the treatment of thoracolumbar fractures by pedicle of vertebral arch Tenor system and Harrington device.Methods The clinical data of 35 cases of thoracolumbar fractures were analyzed retrospectively, 23 of them were operated with Harrington device and 12 with Tenor system.Results These cases were followed up for 6 months to 2 years. The fractures treated with Tenor system got satisfied reduction, reached about 96 7% of normal vertebral height. The deformities were completely corrected and complications were significant lower than those derived from Harrington device treatment, and no delayed neurologic damage was found. Most cases got reduction with one to three Frankel grades of neurologic recovery.Conclusion The Tenor pedicle system is more effective in treatment of thoracolumbar fracture than non-pedicle device. It can get satisfied fracture reduction , reliable fixation and less complications.

Objective To compare the advantages and disadvantages of non-irrigation and irrigation in the surgical approach of chronic subdural hematoma (CSDH),thus to provide reference for clinical treatment of CSDH.Methods Clinical data of 102 patients with CSDH were retrospectively analyzed.According to the different operation methods,the patients were divided into the non-irrigation group(52 cases) and the irrigation group(50 cases).The blood loss during the procedure,operative time,length of stay and postoperative complication rate between the two groups were compared,and the causes of postoperative complications were analyzed.Results The blood loss during the procedure,operative time and length of stay in the non-irrigation group were (6.73 ± 1.17) mL,(15.06 ± 2.64) min and (10.74 ± 2.20) d,respectively,which in the irrigation group were (19.52 ± 3.18) mL,(38.54 ± 6.95) min and (10.44 ± 2.07)d,respectively,there were statistically significant differences between the two groups in the blood loss during the procedure and the operative time (t =-27.11,-22.72,all P ＜ 0.05),there was no statistically significant difference between the two groups in the length of stay (t =0.70,P ＞ 0.05).The incidence rates of postoperative complication in the non-irrigation group and irrigation group were 8.00％ and 7.69％,respectively,there was no significant difference between the two groups (x2 =0.003,P ＞ 0.05).Conclusion Each of the two methods has its own advantages and disadvantages in the treatment of CSDH.Compared with burr hole irrigation,burr hole non irrigation has the advantages of less blood loss and shorter operative time.However,burr hole non-irrigation is more likely to suffer serious complication.We should select suitable surgical approach by the specific circumstances of the patients.The causes of postoperative complications of CSDH are varied.In particular,there is an important relationship between the non-standard operation and postoperative complications.

Objective To detect the expression of the tryptase in the plasma,and study the meaning in brain traumatic patients.Methods There were two groups:the brain traumatic group(40 patients)and the control group (20 health people).The content of plasma tryptase was determined by fluorescence enzyme immunoassay..Results The level of plasma tryptase had no statistical significance in control group(2.97 ±1.05)μg/L compared with the brain traumatic group(3.03 ±1.39)μg/L,however there had statistical significance comparing with sever brain traumatic patients(3.84 ±0.52μg/L)(t =3.32,P <0.05).4 cases of death in patients with severe head injury group content of tryptase (5.85 ±1.05)μg/L,which was significantly higher than the group of 16 cases of injury in severe head injury after 2 months still alive with the content of serum tryptase (2.49 ±0.52)μg/L,the difference was statistically significant (t =8.13,P <0.01).Conclusion The plasma tryptase level in sever brain traumatic patients increased significantly,and might be of importance for treatment strategies and prognosis.

Objective To investigate the satisfaction degree of students and teachers towards PBL teaching method and to provide a basis for continuing to carry out PBL teaching.Methods Anonymous questionnaires of PBL teaching method in teaching plan and procedure was conducted among 49students (2008 grade) and teachers.SPSS 13.0 software package was used to analyze the data and P ＜ 0.05 stands for statistically significant differences.Results Both students and teachers were satisfied.Teachers' evaluation scores of 17 projects were all higher than 4 points and students' evaluation scores of 13 projects were all higher than 4 points; scores for the rest projects were all higher than 3 points.Conclusions PBL teaching method combined with stomatology is effective and measures should be taken to strengthen the management of PBL teaching in stomatology.

Objective:To compare the clinical efficacy and safety between spleen total resection splenectomy and spleen partial splenectomy in the treatment of patients with traumatic spleen rupture.Methods:84 cases of patients with traumatic spleen rupture from March 2013 to March 2016 were selected and divided into two groups.42 cases in the spleen total resection group were treated with spleen total resection,while the other 42 cases in the spleen partial nephrectomy group were treated with spleen partial splenectomy.The operative effect,the levels of platelet count,serum IgA,IgG,IgM,CD3 +,CD4 +,CD8 +,CD4 + / CD8 + levels before and after treatment,the incidence of complications were compared between two groups.Results:The intraoperative blood loss,exhaust time,length of hospital stay of spleen partial nephrectomy group were shorter than those of the spleen total resection group,but the operation time of research group was longer than that of the spleen total resection group (P＜0.05).The platelet count,CD8+ of spleen partial nephrectomy group were lower than those of the spleen total resection group (P＜0.05).The IgA,IgG,IgM,CD3+,CD4+,CD8+,CD4+/CD8+ of spleen partial nephrectomy group were significantly higher than those of the spleen total resection group (P＜0.05).The incidence rate of complications in the spleen partial nephrectomy group was lower than that of the spleen total resection group (P＜0.05).Conclusion:Spleen partial splenectomy was more effective than spleen total resection in the treatment of traumatic splenic rupture,which had little effect on the platelet and immune function.

BACKGROUND: Some researches suggest that placenta is regarded as a new source of mesemchymal stem cells (MSCs). Adherent cells derived from placenta tissue have similar morphological characteristics and surface markers to MSCs; meanwhile, they can differentiate into osteoblasts and nerve cells.OBJECTIVE: To find a new source and way for MSCs separation.DESIGN: Observational study.SETTING: The Third People's Hospital of Wuxi.MATERIALS: Placenta was sourced from cesarean in our department of obstetrics and gynecology and provided confirmed consent from the relatives and ethics committee. The main reagents contained culture medium, sABC kit, DAB staining kit, caprine-anti-rat FITC, recombinant human basic fibroblast growth factor (rh-bFGF), rabbit-anti-human glial fibriliary acidic protein (GFAP).METHODS: The experiment was carried out in the Laboratory of Cells & Molecular Biology, the Third People's Hospital of Wuxi from May 2005 to August 2006. Placenta entity tissue was digested, adherent cells were cultured, morphological characteristics were observed, and growth curves of placenta-derived multipotent cells (PDMCs) in various generations were drawn. On the 1st and 7th days, supernatant was derived from cells in primary culture to measure content of β-glycerophosphate disodium (β-HCG) with chemiluminescence technique. In addition, expression of surface antigen and differentiating potency were detected at the same time. At 24 hours (at phase of nerve cells) and 2 weeks (at phase of osteoblasts) after inducible differentiation, cultured cells were dealt with routine immunocytochemical stain, and then they were observed under routine microscope or fluorescence microscope.MAIN OUTCOME MEASURES: ① Morphological indexes and growth curves of PDMCs; ② measurement of β-HCG in supernatant of PDMCs and expression of antigen of PDMCs with flow cytometer; ③ immunohistochemical analysis of PDMCs during and after inducible differentiation.RESULTS: ① Primary culture of PDMCs: After digestion of placenta tissue, a few of adherent cells were obtained and gradually formed thin and flat monolayer cells two weeks later. The monolayer cells grew like whirlpool or cluster. With the increase of cell density, soma was slenderer and slenderer and like fibroblast. ② Growth curves of PDMCs: Growth latency ranged from 2 to 8 days after cell inoculation. During this period, adherence was observed gradually but not obviously amplified. Eight days later, cells entered log growth phase. During this period, proliferation was active and cell process surrounding extended under phase contrast microscope. A lot of MSCs were observed in division phase between two nuclei. In addition, density was increased and cells connected to each other. Within 11-14 days, growth curve gradually entered platform phase. MSCs covered the bottom of bottle, cells slowly expanded, and primary culture stopped. ③ Measurement of β-HCG: Expression of β-HCG was not detected in supernatant at two time points. ④ Characteristics of surface antigen of PDMCs: PDMCs could express CD29, CD44 and CD105, but not CD34, CD45,CD19 and CD106. ⑤ Inducible differentiation of PDMCs: At 24 hours after inducing to nerve cells, form of PDMCs obviously changed, soma rebounded, refraction of nucleus was partially reinforced, structure was similar to dendrite and axis-cylinder, and positive neuro-specific enolase and GFAP were observed after staining.CONCLUSION:Placenta tissue contains PDMCs whose morphological function is similar to MSCs; in addition, placenta is regarded as an effective source of MSCs.

BACKGROUND: Multipotential mesenchymal stem cells (MSCs) can be differentiated into bone, fat, cartilage, muscle and endothelial cell at the specific conditions, so it draws great attentions of the related study.OBJ ECTTVE: To establish the method of in vitro culture and expansion of human umbilical blood-derived MSCs and investigate their influencing factors.DESTGN: Randomized and controlled trials.SETTING: Department of Tissue Engineering and Cell Biology in Wuxi Third People's Hospital.MATERTALS: Seventy cases of human umbilical cord blood (HUCB) of healthy neonates (selected from Parturient Room, Department of Gynecology and Obstetrics, Wuxi Third People's Hospital, with the informed consents of the puerpera and their relatives), 40 mL each; dulbecco's minimal essential medium (DMEM) of low glucose (LG) at the dose of 50 g/L, fetal bovine serum (FBS), mycillin and trypsin (all were from Gibco), FITC-labeled mice anti-human CD29, CD105 and CD106 (Ancell) monoclonal fluorescent antibody, PE-labeled mice anti-human CD34, CD44, CD45, CD19, HLA-DR (Immunotech)monoclonal fluorescent antibody, Ficoll separation medium (Pharmacia).METHODS: The experiment was carried out in Department of Tissue Engineering and Cell Biology in Wuxi Third neonatal cord blood were anticoagulated by 25 u/mL heparin, and 60 cases of isolated HUCB mononuclear cell were precipitated and suspended by cell culture fluid (50 g/L DMEM-LG + 50 g/L FBS + 10 g/L mycillin), while other 10 cases were purified with DMEM of high glucose (100 g/L) to produce adherent layer. Once the cells reached fluences of FBS coating on adhesive rats of MSCs: Some of the purified HUCB MSCs at logarithmic phase were growth curve of MSCs: The growth of cells were observed by phase contrast microscope (OLYMPUS CK40), and were detected by EPICS-ALTRA flow cytometry.FBS coating and without FBS coating at the different time points, and their adhesive rates.RESULTS: All 70 cases of HUCB MSCs were adopted in this study, however, 10 samples failed to obtain the ideal MSCs cord blood MSCs after culture. During 2 weeks of culture, primary cells began to double for 3-4 days and reached conand CD105, but did not express antigens CD34, CD45, CD106 and HLA-DR, which was identical to surface labeling of those without FBS coating (P＜0.01).CONCLUSrON: MSCs in HUCB can be cultured and expanded in vitro, and are regarded as an alternative source of MSCs for experimental and clinical applications. Moreover, high glucose can depress the growth and proliferation of MSCs in HUCB.