BACKGROUND: Soft tissue filling is a problem in clinic. It has been proved that ultrasonic wave-treated mature adipocytes cannot be used for cell transplantation.It is hopeful to solve the problem with adipose tissue engineering. OBJECTIVE: To observe the effect of ultrasonic wave on the in vitro culture and proliferation of preadipocytes, and validate the possibility of preadipocyte as seed cell in adipose tissue engineering following ultrasound-assisted liposuction. DESIGN: Controlled observation experiment. SETTING: Department of Plastic Surgery, Rennin Hospital, Wuhan University. MATERIALS: This experiment was carried out in the Medical College of Wuhan University from July 2003 to September 2004. One local 3-month-old hybridized pig was provided by the Experimental Animal Center of Medical College of Wuhan University. After the pig was anesthetized with ketamine, an area of 10 cm×20 cm was labeled on both sides of back respectively, and the tissue in the labeled area was harvested. The tissue on the right side served as experimental group and that on the left side served as control group. METHODS: The tissue of the expedmental group was pretreated for 8 minutes by ultrasonic wave with the energy of 3W/cm2. Under aseptic condition, the skin layer was open, and 100 g subcutaneous adipose tissue was resected from each side and then placed in prepared container containing cold D-hanks solution for later use. The ultrasonic wave-treated porcine preadipocytes of adipose tissue of experimental group were isolated and cultured in vitro, and the number and lipid content of preadipocytes were measured every other 2 days. Results were presented as the mean val ue of the number of cells of three wells. In the control group, pretreatment of ultrasonic wave was omitted. The growth curves of two groups were drawn. Intracellular adipose content was measured by oil red O staining. Absorbance (A) was measured with spectrophotometer (HITACHI G2000), which was regulated at the wavelength of 510 nm. MAIN OUTCOME MEASURES: ① Morphological observation of culture of porcine preadipocytes. ②The growth curves of experimental and control groups. ③3 Change in the lipid content of experimental and control groups.RESULTS: ① Cells in the control group adhered to the wall within 6 to 24 hours, and those in the experimental group basically adhered to the wall after 1 to 2 days, and there were many non-adhesive cells in the exoerimental group. Accumulation of adipose granules in the cells was found in the control group after 4 days and that was found in the experimental group after 6 days. On day 12, the cells in the control group basically differentiated into the cells with single lipid drop or multiple lipid drops, and a small amount of natant mature adipocytes were found, while few cells with lipid drop were found in the experimental group. ②Under the condition of the same amount of cells, the cell doubling time of experimental group was about 72 hours and that of control group was 36 hours. After 9-day culture, the number of cells in the experimental group and control group was 13×104 cells/well and 18×104 cells/well, respectively. The rate of cell proliferation of experimental group was very slow. ③Lipid of the experimental group appeared on the 6th day after culture, and that of the control group appeared on the 4th day after culture. The peak value of A was 0.32 and 0.68 in e penmental group and control group respectively.CONCLUSION: Ultrasonic wave has obviously inhibitory effect on the proliferation and differentiation of preadipocytes.The preadipocytes treated by ultrasonic wave are not suitable as seed cells.

Hijack terror is one of the most common terror attacks. In this paper, the definition, the present condition and future development of hijack terror are elucidated. Hijack terrorism may seriously endanger human lives, result in great economic loss, create social panic and political tumult. The medical succor for hijack terror includes treatment and transportation of the wounded, sanitation for epidemic prevention, protection from harmful agents, and medical health care. The challenges confronting medical support for hijach terror are its unexpectedness and abruptness, complexity, and arduousness in its implementation. In order to negotiate these challenges, it is imperative that a flawless counterplan should be worked out, a highly efficient counterterrorism medical organization should be established, intense emphasis should be paid in the studies of anti-neuclear, antimbiological and anti-chemical warfores, and education for psychological prepuredness should be stressed.

Objective To evaluate the feasibility and safety of the treatment for ventricular septal rupture (VSR) by transcatheter interventional therapy coordinated with percutaneous coronary intervention (PCI) or percutaneous transluminal coronary angioplasty (PTCA).Methods Six patients who suffered from cardiogenic shock due to acute myocardial infarction (AMI) and VSR were given intra-aortic balloon pump (IABP) and mechanical ventilation support. After three weeks, they underwent transcatheter closure of ventricular septal rupture with Amplatzer ventricular septal occluder, and at the same time PCI and PTCA were performed. Results All the patients successfully underwent ventricular septal defect occlusion, including two cases of residual shunt. Segmental stenosis of the middle of anterior descending branch and circumflex artery were noted in three patients, and then stent implantation was used. The other three cases had multi-vessel lesion but were without stent implantation. All the patients with stent implantation survived and discharged. Conclusions It is safe and feasible to apply transcatheter closure of ventricular septal rupture with Amplatzer ventricular septal occluder, coordinated with PCI and PTCA for patients with acute myocardial infarction complicated with ventricular septal rupture.

Objective To investigate the effect of doxycycline on the Th1/Th2 cell balance in experimental allergic encepha-lomyelitis(EAE)rats.Methods Forty female Wistar rats were randomly divided into the EAE control group,low,medium and high does DOX treatment groups,10 cases in each group.The onset situation in rats was observed.The IL-4 and IFN-γlevels secreted by peripheral blood mononuclear cells (PBMC)at the peak stage were detected.The levels of IL-1β,IL-10,TNF-αin brain tissue,and the albumin content in cerebrospinal fluid and serum were detected.The QA value was calculated.Results In each DOX group,the clinical symptoms of rats were alleviated compared with the EAE control group.In each DOX group,the PBMC secreting IFN-γlev-el and IFN-γ/IL-4 ratio in the onset peak stage were lower than those in the EAE control group,while the IL-4 level was higher than that in the EAE control group(P <0.01).Compared with the medium-dose DOX group,the increase of IL-4 level in the high-dose DO group was unapparent(P >0.05),but the difference between other DOX groups had statistical significance(P <0.01).The IL-1βand TNF-αlevels of brain tissue and QA value during onset peak stage in various doses DOX groups were decreased compared with the EAE control group,while the IL-10 level was increased compared with the EAE control group(P <0.05).With the DOX dose increasing,the levels of IL-1β,TNF-αand QA value in various doses DOX groups became lower,the IL-10 level became high-er,there was statistically significant difference among various doses DOX groups (P <0.05 ).Conclusion DOX can obviously alle-viate the clinical symptoms of EAE rats,its mechanism may be related with that DOX could decrease the level of Th1 cytokine and increase the level of Th2 cytokine,correct the Th1/Th2 cell balance,thus protect the blood brain barrier(BBB).

Objective To explore the distributions of genotypes of HPV infection in CIN and cervical carcinoma. Methods Cervical exfoliated cells were collected from 365 patients with abnormal cervical histology , and subjected to genotyping assay. Results The most prevalent HPV types were 16, 18,52, 58 and 33. The prevalence ratio of HPV 33,52,58 was signi cantly lower in squamous cell carcinoma. Multiple infections decreased from CIN II/III to cervical cancer. Conclusion Besides HPV 16/18, the 52/58/33 subtypes are also important in the development of cervical cancer.

Objective To describe the natural history of cervical intraepithelial neoplasia(CIN)Ⅰand the biologic factors associated with the progression of CINⅠ and to analyze the predictive values of p16INK4a protein for the progression of CINⅠ. Methods From August 2010 to July 2013, 104 patients referred for abnormal cytology [≤ low-grade squamous intraepithelial lesion (LSIL); including negative for intraepithelial lesion or malignancy (NILM), atypical squamous cells of undetermined significance (ASCUS), LSIL] and high-risk (HR) HPV positive,and were diagnosed CINⅠ by colposcopy-assisted biopsy and followed at 1-year intervals in the First Affiliated Hospital of Nanjing Medical University. In order to analyze the relationship between the progression of CINⅠ with clinical biologic factors, including patient age, cervical cytology before colposcopy, loads of HR HPV, HPV L1 capsid protein, p16INK4a protein,χ2 tests was used to compare the different frequencies of factors in groups of progressed and persisted/regressed CINⅠ, then five factors with progressed CINⅠwere processed into binary logistic regression analysis. Results (1) In the first year of follow-up, among 104 patients(including 15 cases NILM,78 cases ASCUS,11 cases LSIL), 52 cases of them were NILM and HR HPV negative, 30 cases were negative for intraepithelial lesion, 10 cases were CINⅠ, 5 cases were CINⅡand 7 cases were CINⅢ. In total, 82 cases (78.8%,82/104) cases had regressed, 10 cases (9.6%,10/104) persisted, 12 cases (11.5%,12/104) progressed [including 5 cases (4.8%,5/104) progressed to CIN Ⅱ, 7 cases (6.7%,7/104) progressed to CIN Ⅲ, none progressed to invasive cancer]. (2) All patients, 88 cases of them accepted immunohistochemical detection the expression of p16INK4a protein. The result shown that 30 cases (34%,30/88) were positive and 58 cases (66%,58/88) were negative. And 94 cases accepted immunocytochemical detection the expression of HPV L1 capsid protein, 49 cases (52%,49/94) were positive and 45 cases (48%,45/94) were negative. (3) Univariate analysis showed that age of the patient, loads of HR HPV, cervical cytology before colposcopy and the expression of HPV L1 capsid protein were not risk factors of the progression of CINⅠ(all P>0.05) except for the expression of p16INK4a protein (P<0.05). Multivariable analysis found that p16INK4a protein positive was associated with progression of CINⅠ(OR=5.1,95%CI:1.162-22.387,P=0.031). (4) Thirty-one cases were p16INK4a protein positive, 8 cases (27%,8/30) of them progressed,while 4 cases (7%,4/58) of 58 cases with p16INK4a protein negative progressed,in which there were significant difference (P<0.05). The sensitivity was 75%, the specificity was 71%, the positive predictive value was 27%and the negative predictive value was 93%for progression to CINⅡ-Ⅲof p16INK4a protein staining. Conclusions The progression rate of CINⅠwith abnormal cytology (≤LSIL) and HR HPV positive was lower, and there was no progression to invasion at 1-year intervals. Immunostaining of p16INK4a protein as the risk factors of CINⅠprogression could have a role in prediction of CINⅠand the management of high-risk CINⅠ.

Objective To explore the effect on job competency by reforming Fundamental Nursing courses among undergraduate nursing students.Methods 86 nursing undergraduates in Grade 2011 were recruited as the control group receiving conventional teaching method,while 119 students in Grade 2012 were recruited as the experimental group receiving reform of fundamental nursing courses.The teaching outcomes were evaluated by using Job Competence of Nursing Students Evaluation Form and Medical education environment measurement table.Results The reform of fundamental nursing courses had significantly elevated all the aspects of professional competency among nursing undergraduates (P＜0.01),including personal traits,clinical ability,communication,critical thinking,specialty construction[(35.71 ± 3.82) vs.(33.41 ± 4.77),(55.29 ± 8.29) vs.(43.93 ± 8.68),(22.19 ± 2.71) vs.(19.88 ± 2.96),(16.83 ± 2.85) vs.(14.37 ± 2.71),(37.78 ± 6.31) vs.(32.42 ± 5.72)].Meanwhile,three dimensions of medical education environment for nursing students were also improved,including the student's perception of the teacher,self academic perception,self social perception [(35.23 ±5.72) vs.(31.28 6..22),(21.42 ±4.19) vs.(23.42 ±3.53),(19.44 ± 3.86) vs.(18.19 ± 3.47),t=-4.523、-3.503、-2.308,P＜0.01 or 0.05)].However,there was no significant differences in the aspects of learning perception,environment perception,and the total score of medical education environment (t=-1.866、0.725、-1.705,P＞0.05).Conclusions The professional competence and parts of the teaching environment around the nursing students have been elevated through the reform of Fundamental Nursing course.The reform also laid a solid foundation for the employment of nursing students.

Protein arginine methyltransferase (PRMT) can methylate a variety of proteins,including histones and non-histones,affecting a variety of cellular processes such as transcription,RNA splicing,DNA repair,and cell cycle control.The activity of PRMT is affected by a variety of regulatory mechanisms,and its abnormal expression plays an important role in the development of the diseases,especially in tumors such as breast cancer and leukemia.PRMT has a unique value in the diagnosis or treatment of tumors.With the further exploration of PRMT methylation mechanism,PRMT selective inhibitors have made some progress.PRMT-specific inhibitors are expected to be targeted drugs for the treatment of tumors.

Aim To study the effect of CSE ( cigarette smoke extract ) on the single-molecule interactional force between thrombomodulin and thrombin by live-cell single-molecule force spectroscopy. Methods CSE was prepared by a previously reported method. The plasmid of TM-GFP was constructed and transfect-ed in COS-7 cells. The expression of TM-GFP was de-tected by fluorescence microscopy and laser scanning confocal microscopy. The transfected COS-7 cells were grouped ( 1 ) GFP -thrombin group ( 2 ) TM-thrombin group ( 3 ) CSE-TM-thrombin group ( 4 ) CSE- GFP-thrombin group. Force measurements with the thrombin modified AFM tips on the living cell surface were car-ried out on PicoSPM II with a Pico-Scan 3000 control-ler and a larger scanner. The force curves measured in living cells were recorded by PicoScan 5 software and analyzed by MATLAB R2009aMetlab. Results The single-molecule binding force of thrombomodulin and thrombin ( TM-Thr ) was determined ( 60. 90 ± 0. 82 ) pN. The binding probability for TM-Thr was about (22. 58 ± 3. 95)%. Antibody blocking binding proba-bility for TM-Thr was ( 2. 58 ± 2. 0 )%. The binding probabilities for GFP-Thr group, CSE-TM-Thr group and CSE-GFP-Thr group were significantly decreased compared with TM-Thr group ( P<0. 05 ) . The mean value of the most probable single molecular interaction force of thrombin/TM-ECD was determined as ( 45. 30 ± 1. 37 ) pN, the binding probability of thrombin and TM-ECD was ( 23. 25 ± 7. 02 )%. When the binding was blocked with the TM-MAb solution, the binding probability decreased to ( 4. 64 ± 2. 31 )%. The bind-ing probability was ( 8. 31 ± 1. 06 )% in the CSE-TM-thr-S group. When further blocked with TM-MAb, the binding probability was ( 5. 17 ± 2. 96 )%. Conclusion CSE significantly decreases the binding probability for TM-Thr to induce intravascular thrombosis.

Objective To investigate the relationship between the helper T cell 17 (TH 17)/ Regulatory T cells (Treg cells) balance in peripheral blood with acute graft-versus-host reaction (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT),as well as the impact of anti-thymocyte immunoglobulin (ATG) on helper T cells in peripheral blood.Method Seventyeight hematologic patients underwent allo-HSCT,conditioning with or without ATG.Ten healthy volunteers severed as a control group.The helper T and regulatory T cells in peripheral blood were detected by flow cytometry.Enzyme-linked irnmunosorbent assay (ELISA) was used to detect serum concentrations of interleukin(IL)-17,IL-21,IL22,IL23,γ interferon (IFN-γ),and transforming growth factor β1 (TGF-β1).Result The percentage of Treg cells,TH17 cells and ratio of TH17/Treg cells in patients without aGVHD showed no significant difference from the healthy controls (P＞ 0.05).As compared with control group and non aGVHD group,the ratio of Treg cells was increased,the percentage of TH 17 cells,and TH 17/Treg cells were significantly increased in 1-2-degree aGVHD group (P＜0.01).With increased degree of aGVHD,the difference as above was more significant in 3-4-degree aGVHD recipients (P＜0.01).In aGVHD group,the IL-17,IL-23,IL-21 and IFN-γ concentrations were higher than the healthy group (P＜0.01) and non-aGVHD group (P＜0.05).Serum TGF-β1 level in aGVHD group was significantly decreased as compared with healthy group and non-GVHD group (P＜0.05),while IL-22 concentrations showed no statistically significant difference among three groups (P＞0.05).In anti-thymocyte immunoglobulin (ATG) pretreatment group,the absolute count of peripheral blood lymphocytes was less than in healthy control group (P＜0.01).In ATG group,the absolute counts of TH1 cells,TH17 cells,CD3+ CD4+ cells and non-TH1/17 cells were less than in non-ATG group (P =0.0000),while the absolute counts of lymphocytes,CD3+ CD4-cells,and TH 1/17 cells were less than in non-ATG group,but there was significant difference (P＞0.05).Conclusion The balance of TH 17/Treg cells and related cytokines were closely associated with aGVHD after allo-HSCT,and ATG influences the reconstruction of TH 17 and Th1 cells at early stage.