Laboratory of electron microscopy is representative of large instruments laboratories in medical research.And the operation and management in this kind of laboratory are different.Case of forty years of operation and management in the medical laboratory of electron microscopy,this paper analyzed and summarized its successful experiences in the research management system,personnel system and the aspect of equipment maintenance management.Provide a management reference for medical research institutions with the laboratory of similar large-scale instruments.

Purpose To prepare 68Ga-NOTA-SPIO as PET/MRI dual mode imaging probe with high sensitivity and resolution,and further evaluate its in vitro and partly in vivo biological properties.Materials and Methods The precursor SPIO-PEG2000-NOTA was prepared and characterized.The precursor was radiolabeled by using one step method to prepare 68Ga-NOTA-SPIO as dual mode imaging probe.The labeling rate of the probe was determined by rapid thin-layer chromatography.Besides,the in vitro stability and lipid water partition coefficient of the probe were evaluated,and its biodistribution in normal mice was also observed.Results The precursor SPIO-PEG2000-NOTA with uniform dispersion and uniform particle size was prepared,and the dual mode probe 68Ga-NOTA-SPIO was synthesized.The labeling rate reached 99％,and the lipid water partition coefficient (Log P) was (-2.60±0.13).The radiochemical purity of the probe was higher than 95％,as it was incubated in the phosphate buffer and fetal bovine serum within 2 hours.The probe was mainly distributed in the liver and spleen of mice,and its clearance velocity in blood was fast.Conclusion The double mode probe 68Ga-NOTA-SPIO synthesized by one step method has high labeling rate with no need of purification,which has good physic-chemical properties and biocompatibility.The probe can be used in the further research of PET/MRI dual modality imaging.

Objective To introduce the technique and experience in reconstruction of posterior crueiate ligament-posterolateral comer(PCL-PLC)injuries with only one allograft of Achilles tendon. Methods The instable knees in 12 cases with PCL injury combined with three degree chronic PLC injury were treated with PCL reconstruction under arthroscope and PLC reconstruction through posterolateral arc incision.Single bundle grafts of PCL reconstructions in tibial and femoral tunnels were fixed by resorption screws.Fibular collateral ligament(FCL)and popliteofibular ligament(PFL)were reconstructed with reforming Larson(?)method.All reconstruction grafts only needed one Achilles tendon as donator.Total op- eration time was 130 minutes including 90 minutes of PCL reconstruction and 40 minutes of PLC recon- struction.Gradual weight loading was allowed after six weeks of bracing.Results Follow-up for mean 12 months(5-24 months)indicated that tibial“step off”reduction was 83%(10/12)and posterior drawer test of 0-1~+ 75%(9/12).Dial sign evaluated that normal external rotation angle was 75%(8/12).Nor- real varus stress test at 30?knee flexion accounted for 83%(10/12).Scores of Lysholm,Tegner and HSS were 90.5,5.1 and 84.5,respectively(P＜0.01=.Conclusion Synchronized reconstruction of PCL and PLC injuries with only one Achilles tendon can obtain satisfactory clinical result,with less expense and shorter operation time.

Immunohistochemistry was used to detect the protein expression of Snail in 5 specimens jusxta-eancerous normal breast tissues, 35 specimens of cancerous tissues without metastasis and 20 specimens of breast carcinoma with lymphonode metastasis. Breast carcinoma cell line MDA-MB-231 was transfected with antisense Snail. Results showed that the expression of Snail protein was significantly higher in breast carcinomas than in their normal tissues. The mRNA and protein expressions of Snail in the breast carcinoma cells treated with antisense Snail was significantly decreased while the E-cadherin protein significantly increased (both P < 0.05). The number of invasive ceils treated with antisense Snail was (10.5±1.3)%, while in treated with EGFP was (68.2±2.1)% (P < 0.05). The abnormal expression of Snail contribute to the invasiveness of breast carcinoma. The antisense Snail could prevent the cells ability to invade in vitro, and the effect is related with the up-regulated E-cadherin protein.

Objective To study the relationship between Couagen Ⅰ,MMP-2,TIMP-2 gene expression and atrial fibrosis during heart failure(HF)in dog.Methods Fourteen dogs were used and randomized into HF induced by ventricular tachypacing and control group.Burst atrial pacing was used to induce atrial fibrillation(AF).And the mRNA and protein level of collagen Ⅰ,MMP-2 and TIMP-2 were detected by RT-PCR and immunohistochemical technique.Tissue samples were stained with Mallory trichrome.Results Left ventricular ejection fraction (LVEF) decreased from (67.4? 6.0)% to (29.2?7.8)%,the inducible rate of AF(7/7 vs 2/7) and sustained AF(5/7 vs 0/7) increased and duration of AF stabeatrial fibrillation(SAF) [(462.12?181.43)s vs(0.57?0.57) s] prolonged significantly in HF group.Atrial fibrous tissue content and atrial size of HF group were significantly greater than the controls dogs(268.8% in lefe atria and 190.3% in right atria).The mRNA and protein level of collagen Ⅰ(56.2% and 132.2% in lefe atria,37.4% and 78.0% in right atria)and MMP-2 (100.0% and 115.7% in lefe atria,65.7% and 96.8% in right atria) increased evidently in both lefe atria and right atria,TIMP-2 mRNA decreased 46.3% in lefe atria and had no change in right atria and that its protein had no change in both atrium,whereas the ratio of MMP-2/ TIMP-2 of mRNA and protein increased markedly in both lefe atria (285.3% and 148.8%)and right atria (106.1% and 134.7%)of HF group.SAF had a positive correlation with fibrosis and the gene level of collagen Ⅰ in lefe atria,the ratio of MMP-2/TIMP-2 had a positive correlation with fibrosis and collagen Ⅰ gene level in lefe atria during HF.Conclusions The changes of collagen Ⅰ,MMP-2 and TIMP-2 gene expression appear to be a molecular mechanism of AF, and the molecular remodeling of collagen Ⅰ induced by regulation unbalance of MMP-2/TIMP-2 appears to be an important mechanism of atrial fibrosis during HF.

Background and purpose:Ionizing radiation(IR) activates the early growth response- I(Egr1) promoter through specific cis-acting sequences termed CArG elements by production of radical oxygen intermediates(ROls).Egr-EG,an expression vector pCIneo containing CArG elements cloned upstream of the cDNA for human recombinant GM-CSF,was used to treat hematopoietic damage due to chemotherapy.Commonly used chemotherapeutic agents can cause tumor cell death by producing DNA damage and generating ROIs.We therefore hypothesized that clinically employed chemotherapeutic agents that increase ROIs could also be employed to activate Egr-EG in a chemoinducible gene therapy strategy.This study was done to explore the chemo-protective effect of the expression of hematopoietic growth factors regulated by Egr-1 promoter on chemotherapy induced damage. Methods:The human GM-CSF cDNA and EGFP cDNA were linked together with internal ribosome entry site(IRES) and then inserted into the eukaryotic expression vector pCI-neo with the Egr-1 promoter(Egr-EG),and was further transduced into human bone marrow stromai cell lines HFCL(HFCL/EG).The HFCL/EG cells were transplanted i.v.into BI6 melanoma in C.B-17 combined immunodeficient(SCID) mice.5-FU was given i.p.on day 3 and 4.The white blood cell amount in peripheral blood,the expression of EGFP and GM-CSF in human stromal cell engrafted in recipient mice were detected by flow cytometry,RT-PCR,Western blot and colony-forming units for granulocytes and macrophages(CFU-GM),respectively.Results:In contrast to the two control groups,HFCL/EG(the Egr-regulatory element-derived expression of GM-CSF gene therapy) resulted in a proportional increase in the number of the white blood cell after chemotherapy,no significant diifferences were found for CFU-GM in bone marrow cells and the inhibition ratio on tumor in recipient mice.Chemotherapy could markedly increase the expression of EGFP and GM- CSF mRNA/protein as compared with that of non-chemotherapy control groups and HFCL group.Conclusion: Chemoinducible GM-CSF gene therapy regulated by Egr-1 promoter can ameliorate the toxic effect on 5-FU chemotherapy-inducible hematopoietic damage.

Objective To explore the feasibility of fishing net repairing transverse fascia method for inguinal hernia (type Ⅰ , Ⅱ ) using laparoscopic surgical procedure. Methods A retrospective analysis of clinical data between the method of fishing net repairing transverse fascia surgery for 145 cases of inguinal hernia (typeⅠ,Ⅱ ) using laparoscopic surgical procedure from May 2004 to May 2008 (laparoscopic group) and the method of open repairing surgery 143 cases (open group) at the same period were conducted. The differences in the operative time, rehabilitation activities time, length of stay, cost of hospitalization and 0comphcations, recurrence rate were compared. Results The laparoscopic group was significantly better in the operative time [ ( 14.8 ± 11.5) min ], found hiding oblique hernia ( 15 cases), rehabilitation activities time[ ( 16.5 ± 14.3) h], use of analgesics(5 cases), scrotal edema(1 case), length of stay[ (4.2 ± 1.5) d], than those of the open group [ ( 37.6 ± 25.4) min, 0, (52.7 ± 12.6) h, 13, 14, ( 8.4 ± 2.6 ) d respectively ] ; but the recurrence rate was no significantly different. Conclusion Method of fishing net repairing transverse fascia for inguinal hernia (type Ⅰ , Ⅱ ) using laparoscopie surgical procedure is feasible.

In recent years, corneal refractive surgery has flourished, but some patients have developed visual adverse symptoms such as decreased nighttime vision, glare, and halo after surgery. As one of the important factors affecting postoperative visual quality, accurate measurement and evaluation of the effective optical area is particularly important. This not only provides guidance for the selection of preoperative surgical methods and the design of optical areas, but also provides better postoperative visual quality for patients. This article reviews the research progress on the design and effective optical area of corneal refractive surgery.

Objective To evaluate the prognosis of full-term neonates with birth asphyxia usingneonatal behavioral and neurological assessment (NBNA). Methods A total of 326 full-term neonateswith birth asphyxia were evaluated using NBNA 12-14 days and 25-28 days after birth. The infants werefollowed up till 24 months old and a developmental assessment was made according to the CDCC Scalesof Infant Development. Results The incidence of developmental retardation (with mental developmentindex ≤69 or psychomotor development index ≤69) was significantly higher in the infants with NBNAscores ≤35 than in those with NBNA scores >35. Conclusion The NBNA score provides a valuablereference for prognostic evaluation of the full-term neonates with birth asphyxia, and may serve as asensitive indicator for cerebral lesions.

Objective: To investigate the pedestrian traffic safety behaviors of primary and secondary school students in Xiacheng District of Hangzhou,and to provide reference for formulating traffic safety strategy. Methods: In October 2015,nine campuses of six schools in Xiacheng District were selected to observe the pedestrian traffic safety behaviors of the primary and secondary school students walking to and from school,such as taking the zebra crossing,observing the traffic condition,running and playing with cellphones when walking across the road. Results: Totally 2 585 students were investigated,and 2 580 students were valid（99.81%）. When walking across the road,1 887 people took the zebra crossing,accounting for 73.14%;71.89% of boys and 74.51% of girls took the zebra crossing,and there was no significant difference（P>0.05）;64.76% and 81.32% of the students took the zebra crossing on the way to school and home,with a significant difference（P<0.05）. When walking across the road,1 683 students people observed the traffic conditions,accounting for 65.23%;63.61% of boys and 67.02% of girls observed the traffic conditions,and there was no significant difference（P>0.05）;64.05% and 66.39% of the students observed the traffic conditions on the way to school and home,with a significant difference（P<0.05）.There were 362 students running across the road, accounting for 14.03%;15.31% of boys and 12.62% of girls ran across the road,and there was no significant difference（P>0.05）;9.26% and 18.68% of the students ran across the road on the way to school and home,with a significant difference（P<0.05）. There were 53 students playing with cellphones when crossing the road,accounting for 2.05%;2.29% of boys and 1.79% of girls played with cellphones,and there was no significant difference（P>0.05）;1.41% and 2.68% of the students played with cellphones on the way to school and home,with a significant difference（P<0.05）. Conclusions Primary and secondary school students have dangerous behaviors when walking across the road. We should focus on the intervention of not taking the zebra crossing and not observing traffic conditions when crossing the road on the way to school,and playing with cellphones on the way home.