Embryonic stem cell (ESC) is a kind of stem cell with self-renewal capability and developmental luripotency,which is at the early stage of embryonic development,and it is the original cell of various kinds of tissue and organ.Under certain conditions,ESCs can maintain an undifferentiated state,long-term surviving and infinite breeding state in vitro.The research of tberapeutic effect of ESCs in animal disease model,especially in ouse,has made great progress.On the basis of this research,researchers attempt to use ESCs for the clinical therapy of various diseases,and investigate the feasibility and security of this kind of therapy.Currently ESCs have been used in the therapy of diabetes mellitus,Parkinson's disease and cardiovascular injury.Even though many problems of technique and ethics should be done,we believe that ESC therapy will be a promising method and used in linical practice if we make clear the mechanism of ESC directional differentiation,establish an optimal culture system,and solve the problem of oncogenicity.

Objective To evaluate the clinical results of surgical treatment of isolated fractures of the sustentaculum tali of calcaneus via medial approach.Methods The data of 23 patients with isolated fractures of the sustentaculum tali of calcaneus was retrospectively analyzed who were treated with open reduction and internal fixation with cannulated screw or Kirschner wire via medial approach from September 2006 to March 2011.There were 19 males and 4 females,with an average age of 26.3 years (range,17-41 years).Associated injuries included 4 cases of talus fracture,4 of metatarsal fracture,and 3 of cuboid fracture.The functions of hiudfoot were assessed by American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot scores pre-operation and post-operation respectively.Results Fifteen patients got followed up with an average 20.5 months (10-56 months).Thirteen patients were rated as good,2 as excellent,and the excellent and good rate was 100％(15/15).All the fractures were stabilized reliably,and got clinical union with no obvious complications occurred.Time of fracture union was 8-10 weeks,with an average of 8.5 weeks.Three patients felt mild transient pain during the recovery of walking,but their pain disappeared quickly after physical therapy.No patients developed wound infection,nonunion and other complications.Conclusion For isolated fractures of the sustentaculum tali of caleaneus with articular surface displaced greater than 1 mm or involving the articular surface of middle subtalar joint,open reduction and internal fixation operation via medial approach under direct visualization is recommendable.

Objective To establish characteristic spectrum of ginsenosides in Panax quinquefolium from Liuba. Methods Using C_(18) solid phase-extraction cartridges, main ginsenosides from 40% ethanol extracts of P. quinquefolium were purified. Then the samples were analyzed by HPLC-EMS. Results(From the) total ion spectrum of P. quinquefolium, 3 stronger peaks were selected. Based on them, characteristic corresponding spectrum of ginsenosides in P. quinquefolium from Liuba was established. Conclusion This method has reliable reproducibility and precision. Its simple pretreatment, easily operation, and rapidly analytic procedure show that this method is suitable for identifying P. quinquefolium.

Objective To express and detect the antigenicity of human laminin alpha4 LG3-4 module (hLN?4LG3-4) protein by gene engineering techniques. Methods The cDNA encoding hLN?4LG3-4 was amplified by RT-PCR from human placenta, then inserted into pMD-18T vector by T/A cloning and sequenced. Prokaryotic expression vector pET-28a-LG3-4 was constructed by recombinant DNA technique. The hLN?4LG3-4 fusion protein expressed in BL21(DE3)/pET system was identified by SDS-PAGE, purified by Ni-NTA resin, and assayed by Western blotting. Results The cDNA fragment of hLN?4LG3-4 was cloned successfully. While BL21 (DE3)/pET28a-LG3-4 bacterium was induced with IPTG, a new protein band with a relative molecular weight of 44 000 was shown on SDS-PAGE profile. hLN?4LG3-4 fusion protein of high purity (95%) was obtained and specific protein band was detected by Western blotting. Conclusion The hLN?4LG3-4 fusion protein was successfully expressed.

Objective To discuss the application of spectral CT imaging in evaluating the early therapeutic response of 125I seed interstitial brachytherapy for pancreatic carcinoma in experimental nude mice.Methods BxPC-3 human pancreatic cancer cell xenografts were subcutaneously inoculated at the dorsal part of the limbs in 16 BABL/c nude mice. When the tumor developed to the size of 1-1.5 cm, the tumor-bearing mice were randomly divided into the study group (n=8,receiving implantation of 125I seeds of 1.0 mCi) and the control group (n=8,receiving implantation of ghost shell particles of 0 mCi). Two weeks after the procedure, plain spectral CT scan as well as enhanced multiphase (10 s, 25 s and 60 s) spectral CT scan were performed to obtain multi-phasic image series. The contrast-to-noise ratio (CNR) and iodine concentration (IC) inside the lesions were determined, and the normalized iodine concentrations (nIC) of tumor were thus obtained. Immunohistochemical staining of tumor was used to measure microvessel density (MVD) within the tumor;the correlation between nIC and MVD was analyzed. Results The lesion's nICs measured on each of three phases (10 s,25 s and 60 s) in the study group were lower than those in the control group, the differences were statistically significant (P<0.05). MVD count in the study group was lower than that in the control group, the difference was statistically significant (t=5.957,P<0.01). A parallel linear correlation existed between nIC obtained from each phase of three phases and MVD count (r=0.63,P<0.000 1;r=0.51,P=0.002;and r=0.48,P=0.001 7 for 10 s, 25 s and 60 s phases respectively). Conclusion Spectral CT imaging is an effective method for evaluating the therapeutic effect of 125I seed interstitial brachytherapy for pancreatic carcinoma of experimental mice.

Objective To retrospectively analyze imaging features of multi-slica computed tomography (MSCT) and clinical characteristics of male patients with pancreas solid pseudopapillary tumor (SPT) and compare with that of female patients.Methods From November 2000 to October 2012,clinical data of 18 male patients and 94 female patients underwent MSCT examination and pathological diagnosed as SPT were retrospectively analyzed.Clinical characteristics and imaging features of MSCT of male and female patients with SPT were analyzed,which included lesion location,size,shape,encapsulation,calcification,internal composition,density and enhancement pattern of tumors.Rank sum test or x2 test was performed for statistical analysis.Results The median age of male patients with SPT was significantly older than that of female patients (39.0 (15.0,67.0) years vs 27.5 (11.0,63.0) years; U=2.865,P =0.005).There were no significant differences in clinical manifestation,lesion location and composition ratio of benign and malignant tumors between male and female patients with SPT (all P＞0.05).Imaging of MSCT indicated that the median maximum tumor diameter of male patients with SPT was significantly shorter than that of female patients (5.1 (1.0,11.6) cm vs 7.9 (2.5,18.7) cm; U=3.161,P=0.002).Solid tumors were more common in male patients compared with female patients (9/18 vs 5.3％ (5/94) ; x2 =30.606,P＜0.01).Conclusions The imaging features of male patients with SPT are different with those of females.For pancreas lesion in males,if imaging of MSCT shows more solid composition,small lesion and with typical enhancement patterns of SPT,the possibility of SPT should be considered.

Objective To investigate the inhibitory effect of carbon-coated iron nanoparticles carrying cisplatin on the growth of NCI-H446 lung cancer cells and expressions of Caspase 3 and Survivin mRNA.Methods NCI-H446 lung cancer cells were treated with iron-carbon nanoparticles and/or cisplatin.The cell viability was detected by MTT method,and the mRNA expressions of Caspase 3 and Survivin were measured with RT-PCR.Results Cisplatin could inhibit the growth of NCI-H446 lung cancer cells,and the inhibitory effect was stronger when it was combined with the iron-carbon nanoparticles.The cells had apoptosis.The mRNA expression of Caspase 3 of NCI-H446 lung cancer cells was remarkably enhanced after treatment with iron-carbon nanoparticles combined with cisplatin,while the mRNA expression of Survivin was notably weakened (P＜0.05).Conclusion Carbon-coated iron nanoparticles carrying cisplatin could significantly increase the chemotherapy sensitivity of cisplatin on NCI-H446 lung cancer cells and enhance the therapeutic efficacy of chemotherapy drugs.

Objective To identify the changes of DNA methylation profile in the process of malignant transformation of BEP2D cell induced by α particles.Methods The genomic DNAs were isolated from the malignant transformation BERP35T4 cells and immortalized human bronchial epithelial cell line BEP2D.Genomic DNAs were digested by MseI and ligated of PCR linkers.Methylated DNAs were digested by BstUI and amplified by PCR.The methylated DNA probes were prepared by labeling with Cy3 and Cy5 fluorescence dyes individually and hybridized to the methylation CpG-Island microarray.The hybridization results were scanned and analyzed.Intensity values were quality controlled and normalized.The normalized data were used to identify the differentially expressed genes based on a 1.5 fold difference of the expression level.Results There were 16 genes which showed changes of methylation level in malignant transformation BERP35T4 cells, 9 of them were hypermethylation and 7 were hypomethylation.These genes were including the SKIP gene, PPP3CC gene, MAP2K6 gene, KIR2DL1 gene, KIR2DL4 gene, KIR3DP1 gene, ZNF493 gene, ZNF100 gene, NKX2-5 gene, TFAP2D gene, DR1 gene, KCNJ16 gene, CCDC18 gene, FNBP1L gene, IRX4 gene, EPB41L3 gene, TCP10 gene and so on.Conclusions The DNA methylation might have effects on ionizing radiation drived tumorigenesis.

Objective To explore the expression of HCCR, pERK and pELK1 in gastric cancer tissue and para-carci-noma tissue, and to explove its correlations. Methods In 60 human gastric cancers and their corresponding paraneo-plastic tissuses,the expression of HCCR, pERK and pELK1 were detected by immunohistochemistry. Results The ex-pression of HCCR was 75.0%(45/60), pERK, and pELK1 were 73.3%(44/60) and 71.7%(43/60), respectively, and the expression of proteins in gastric tumor were higher than that in paraneoplastic tissues (P<0.05). Conclusion The HCCR, pERK and pELK1 are higher expressed in tumor tissues, availably as a auxiliary index for clinical immunohis-tochemistry in gastric cancer.

Osteochondral lesion of talus (OLT) is a foot and ankle disease characterized by ankle pain, which may impact the joint function and life quality. If managed improperly, it may lead to a further ankle arthritis, severely compromising the prognosis. The therapeutic effect of conservative treatment for OLT is still uncertain. Surgery is still the main treatment modality for OLT with various techniques. However, the optimized surgical technique is still inconclusive, furthermore, regeneration and repair of cartilage after debridement is also a great challenge for the treatment of OLT. Platelet-rich plasma (PRP) with good repair effect on cartilage injury is gradually applied in the treatment of OLT. However, there still lacks the unified understanding of the technique and specification of PRP for the treatment of OLT. Therefore, National Orthopedics Center of Shanghai Sixth People′s Hospital allied Foot Ankle Basic Research & Orthopedics Group, Chinese Association of Orthopedic Surgeons; Foot and Ankle Committee of Chinese Association of Sports Medicine Physicians; and Foot and Ankle Group of Orthopedic Specialized Branch of Shanghai Medical Association to organize related experts to formulate the Expert consensus on platelet- rich plasma treatment for osteochondral lesion of talus ( version2023). Fifteen recommendations were put forward upon PRP preparation, indications, contraindications and treatment methods of PRP for OLT, so as to standardize the PRP treatment for OLT.