This study aimed to investigate the method of conserving the mice skin for long time in vitro,and forty clean male KM mice of 8-week old were divided into two groups by random digit table,namely vitrified skin transplantation group(n =30) and fresh skin transplantation group(n =10) .The skin was vitrified quickly with freezing solution EFS40 by the two-step method,and then was thawed with 0.5 mol/L glucose,which was followed by self-transplanted.The vitrified skin transplantation group was self-transplanted after the skin was vitrified for 2 weeks.A piece of skin which was smaller than the vitrified one was cut off on the other side of the spine,and the thawed skin were laid smoothly to coincide with the edge of the cut skin which was sutured then.In fresh skin transplantation group,fresh skins were prepared to put in Dulbecco phosphate-buffered saline for 15 minutes and then be transplanted to original position.The outcomes of transplantation were valuated on day 14 and day 28 after transplantation.And the results indicated that the survival rate of the fresh skin transplantation group was 100%(10/10) .The survival rate of the vitrified skin transplantation group was 77%(23/30) on day 14 and 70%(21/30) on day 28,respectively.But the surfaces of the mice skin and part of the hair follicle cells were damaged during vitrification.However,the method of vitrifying the mice skin quickly in this paper may be used to conserve the skin in vitro.

Fetal Blood ; Humans ; Infant, Newborn ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; congenital ; diagnosis

Objective To study clinicopathological characteristics of hereditary nonpolyposis colorectal carcinoma(HNPCC)-associated endometrial carcinoma(EC).Methods Totally 421 EC patients admitted to General Hospital of Tianjin Medical University from 1981 to 2006 were divided into three groups:group A:sporadic EC;group B:familial aggregation of EC;group C:HNPCC-associated EC.Results HNPCC-associated EC accounted for 6.4%(27/421).Mean age at time of diagnosis was 49.7years in group C,earlier than 56.3 years in group A(P=0.004)and 55.2 years in group B(P=0.035).There were 33.3%(9/27)patients with multiple primary carcinomas in group C.It was higher than 14.3%(9/63)in group B and 5.1%(17/331)in group A respectively(P=0.038,P＜0.01).There was no difference among the three groups in histological type or menopausal status(P＞0.05).The numbers of patients with low grade EC in group C(70.4%,19/27)and group B(61.3%,38/62)were more than that in Group A(P=0.013,P=0.023).Prognosis for group C was better than that in group A(P=0.021),and 5-and 10-year survival rate in groups A,B and C was 80%,70%;88%,85%;and 96%,96%respectively.Conclusions Mean age at time of diagnosis is earlier in HNPCC-associated EC than that in sporadic EC;patients with HNPCC-associated EC are more frequently complicated with multiple primary carcinomas and of low grade;prognosis is better.

Objective To explore the methods of labeling exogenous microglia with superparamagnetie iron oxide(SPIO)particles,and to monitor the labeled cells after transplantation into the normal rat and Alzheimer's disease(AD)model rat with MR scanning.Methods Microglia was labeled with SPIO particles by using transfection agent,hemagglutinating virus of Japan envelope(HVJ-E).Then the microglias which were labeled with SPIO were injected into the internal carotid artery of normal rat (n=5)and AD model rat(n=5).Three days after transplantation,follow-up serial T2*-weighted gradient-echo MR imaging was performed at 7.0T MRI system.MR images were correlated with histological findings.Results In the brain of normal rat,the labeled microglias were demonstrated as several dotty signalintensity decrease on T2*-weighted MR images.The dotty spots were sporadic around the brain.Histological analysis showed that most prussian blue staining-positive cells were well correlated with the area where a signal intensity decrease was observed in MRI.MR could detect the signal intensity change caused by a few labeled cells.In the brain of AD model rat,MR scan showed a well-defined hypointensity area in the region of Aβ42 iniection.Signal intensity decrease was not obvious in the region of saline injection.The number of iron-positive cells(454±47)/mm2 at sites of Aβ42 injection was much higher than that(83±13)/mm2 of saline injection(P＜0.05). Conclusion MR can be used as a non-invasive means of detecting transplanted labeled microglia in vivo,with the potential for future clinical application in cell therapy of AD.

ObjectiveTo investigate the effects of vascular endothelial growth factor (VEGF) and dexamethasone on mRNA expressions of surfactant protein B (SP-B) and transforming growth factor-β1 (TGF-β1) of type Ⅱ alveolar epithelial cell (AECⅡ). MethodsAECⅡ were isolated and purified from fetal rat lung tissues,then cultured with different dose of VEGF (25,50 and 100 ng/ml) and dexamethasone (25,50,100 and 200 nmol/ml).The mRNA levels of SP-B and TGF-β1 were detected by real-time quantitative polymerase chain reaction (RT-PCR) and expression of TGF-β1 protein was detected by immunocytochemistry.ANOVAor q-test wasappliedtocompare the difference among groups.ResultsCompared with control group,SP-B mRNA levels in 25 ng/ml VEGF group and 25,50,100 and 200 nmol/ml dexamethasone groups were higher (13.500±3.172,3.547±0.690,5.219±0.782,3.493±0.335,and 3.981 ± 1.133 vs 1.001 ± 0.059,q=-5.286,-4.943,- 7.228,- 9.906 and - 3.525 respectively,P＜0.05) ; TGF-β1 mRNA expression of 25 ng/ml VEGF group,50,100 and 200 nmol/ml dexamethasone group was lower (0.451 ± 0.078,0.579±0.019,0.422 ± 0.020 and 0.769 ± 0.025 vs 1.019±0.226,q=4.110,3.356,4.551 and 1.901 respectively,P＜0.05) ; other groups had no significant differences compared with control group (P＞0.05).Immunocytochemistry showed that the positive rate of TGF-β1 expression in 25 ng/ml VEGF,50,100 and 200 nmol/ml dexamethasone group was 23％,26％,22％ and 29％,respectively,while in the control group,the expression of TGF-β1 was positive in most of the AECⅡ (80％).ConclusionsBoth VEGF and dexamethasone could increase the expression of SP-B at mRNA level at appropriate concentrations.At the same time,the expression of TGF-β1 is inhibited.It is suggested that both VEGF and dexamethasone might increase the mRNA expression of SP-B by inhibiting the expression of TGF-β1.

ObjectiveTo investigate the relationship between the oxidative injury induced by low glucose and mitochondrial membrane potential in HUVEC-12 cells. Methods Human umbilicalvein endothelial cells HUVEC-12 were cultured in low concentration glucose for 4 h.Cell viability of HUVEC-12 cell was assessed with MTT assay.Dihydroethidium (DHE) was used as a reactive oxygen species (ROS)capture, which was detected the mean fluorescence intensity of samples and Rhodamine 123 as a fluorescence detector was to measure the level of mitochondrial membrane potential (MMP) in cells.Results Comparing to HUVEC-12 cells viability in 5.5 mmol/L glucose group (96.80 ±3.20)％, cells exposed to 2.8 mmol/L glucose group (66.40 ± 1.60) ％ and 0 mmol/L glucose group (58.93 ± 1.67) ％ were decreased by 32％ and 40％ respectively (P ＜ 0.01).ROS level of 5.5 mmoL/L glucose group, 2.8 mmol/L glucose group and 0 mmol/L glucose group were 0.59 ± 0.02, 0.74 ± 0.04 and 0.88 ± 0.05,respectivdy, increased by 25％ in cells exposed to 2.8 mmol/L glucose and by 48％ in cells without glucose exposure comparing to 5.5 mmol/L glucose group (P ＜0.01) ; MMP levels of 5.5 mmol/L glucose group,2.8 mmoL/L glucose group and 0 mmoL/L glucose group were 148.83 ± 3.51, 271.07 ± 19.54 and357.74 ±51.32 respectively, increased to 1.8 times in cells exposed to 2.8 mmol/L glucose and to 2.4times in cells without glucose exposure comparing to 5.5 mmoL/L glucose group (P ＜ 0.01).Conclusion Low glucose leads to injury in HUVEC-12 cells, which is probably induced by the oxidative stress via the increasing MMP.

Objective To explore the effects of application of laparoscopic simulators in teaching of plastic surgery. Methods 10 plastic surgeons and 20 standardized training surgical residents with 2 to 4 years' experience were tested about their proficiency in moving beans, pinching, suturing and tying by timing and counting. After they were trained with laparoscopic simulators 3, 6, 9 times with each time for 90 min, tests were taken. SPSS 19.0 was used to make single factor variance analysis of the related data or conduct q test. Result There was significant difference before and after the residents' training of moving beans, pinching, suturing and tying (P<0.05). Less time to finish the operation was needed after training, but after training for 6 times or 9 times, there was no significant difference in operation time (P >0.05). Conclusion Application of laparoscopic simulator training can significantly improve the operation skills of the novices with some clinical experience in the short term, which is conducive to the endoscope assisted breast augmentation surgery, and is worthy of promotion.

Objective To observe the anesthesia and recovery results of sevoflrane and remifentanil combined anesthesia in open or laparoscopic surgeries. Methods 60 cases of ordinary surgeries from the department of gynecology and general surgeries were included in this study with 30 cases in each group. (1)recording total sevoflurane inhalation time, muscular relaxant amount, end tidal sevoflurane concentration;(2)recording BP,HR at 10 min after induction,operation staring and ending,ventilation recovery, opening eye and extubation period;also sevoflurane concentration 5 min after stopping medicine and ventilation recovery;recording time period between surgery ending and autonomous respiration recovery , eye opening and extubation. Results No any adverse events happen in each patient.the sevoflurane inhalation time in open surgery group was (157.20 ± 47.28) min, longer than that of laparoscopic surgeries group (73.50 ± 11.23)min(P<0.05), we had seen no statistical significance in all other index observed (P>0.05). Conclusion Sevoflurane combined remifentanil anesthesia can achieve stable intra-operative maintenance and rapid postoperative recovery quality , we suggest the widespread usage of it in clinic.

By using single spleen colony transplantation technique and sex chromosome typing as natural cytogenetic markers, we have been able to demonstrate that most spleen colony forming cells in the adult bone marrow or in foetal liver of inbred LACA or C57 mice possess the potential of re-establishment of myeloid and lymphoid lines of cells in lethally irradiated mice. However, most of the spleen colony forming cells in the peripheral [blood of normal mice possess weak or no potentiality to proliferate or re-establish hemopoiesis in lethally irradiated mice. Our results support the notion that the CFU-S population in mice is heterogeneous.From the nature of colony forming cells in the peripheral blood of normal mice, we are convinced that the assessment of CFU-S content relying upon the spleen colonies as the unique sign to indicate the self-renewal ability of a spleen colony forming cell is by no means valid inasmuch as some plu-ripotent . hemopoietic progenitors derived from pluripotent hemopoietic stem cells may have the same ability to form colonies in vivo.

Objective To investigate the value of the three-dimensional endorectal ultrasonography (3D-ERUS) in the tumor staging before transanal endoscopic microsurgery (TEM).Methods The clinical data of 30 patients with rectal cancer who underwent 3D-ERUS before TEM at the Nanjing Hospital of Traditional Chinese Medicine from April 2012 to December 2013 were retrospectively analyzed.The accuracy,sensitivity and specificity of the 3D-ERUS were evaluated according to the results of the postoperative pathological examination.The consistency of the results of the 3D-ERUS and postoperative pathological examination were compared by Kappa consistency test.Results Of 30 patients,25 patients in stage T0,3 in stage T1 and 2 in stage T2 were diagnosed by preoperative 3D-ERUS.There were 2 patients (stage pT0) with inflammatory polyp by postoperative pathological diagnosis,6 patients (stage pT0) with tubular adenoma,16 patients (stage pT0) with villioustublar adenoma,2 patients (stage pTis) with carcinoma in situ,2 patients (stage pT1) with tectal adenoma and 2 patients (stage pT2) with rectal adenoma.There were 2 patients with excessive tumor staging by 3D-ERUS,1 patient in stage pT0 was misdiagnosed in stage T1,1 in stage pT1 was misdiagnosed in stage T2 and 1 in stage pT2 was misdiagnosed in stage T1 with insufficient tumor staging.The accuracy of 3D-ERUS in the preoperative tumors staging of TEM was 90.0％ compared with the resuls of postoperative pathological examination.The accuracy,sensitivity and specificity of 3D-ERUS in stage pT0,pTl,and pT2 of TEM were 96.7％,90.0％,93.3％ and 96.2％,50.0％,50.0％ and 100.0％,92.8％,96.4％,respectively.There was a significant difference in the consistency between preoperative 3D-ERUS and postoperative pathological examination (κ =0.685,P ＜ 0.05).Conclusion 3D-ERUS is an accurate clinical method in the preoperative tumors staging of TEM,and can be used as the preoperative assessment for TEM.