Objective To establish the three-dimensional culture model of the human skin squamous-celled carcinoma ,and isolate the highly invasive cell subsets ,and compare their biological characteristics .Methods By using the composite chitosan tissue engi-neering skin ,the three-dimensional culture model of the human skin squamous-celled carcinoma was successfully established , through digestion ,we divided and got the high-invasive squamous carcinoma cells subsets .After that ,the different invasive proper-ties of subpopulations of cells in nude mice tumor effects were compared .Results Different concentrations of cinnamaldehyde and interferon on skin squamous-celled carcinoma cell proliferation inhibition had statistical significance (P<0 .01) ,the different invasive characteristics of skin squamous cells carcinoma between subpopulations of cells proliferation inhibition also was statistically signifi-cant(P<0 .01) ,and the two different invasive properties of subpopulations of cells in the tumor characteristics also had statistical significance(P<0 .01) .Conclusion We established the three-dimensional culture model of the human skin squamous-celled carcino-ma successfully ,by using this model ,we could screen the higher invasive cells subsets .

Objective To investigate the influencing conditions of human epidermal stem cells(ESCs) differentiating into gland-like cells and to identify the induced tubular structure.Methods The ESCs were seeded onto compound polysaccharide shell dermal matrix and collagen gel,adding different concentrations of epidermal growth factor(EGF) and culturing by vertical shaking in vitro,the three dimensional culture and induced directional differentiation were performed.The means of HE staining,immunofluorescence and confocal laser scanning microscope were adopted to observe the conditions,morphology and phenotype change of ESCs directionally differentiating into glandular epithelium-like cells.Results 15-20 ng/mL EGF could induce ESCs in tissue-engineering dermis to grow into dermis and appear the gland-like structure.The HE staining in this structure showed its profile as a single layer with lacune in the middle compartment,eosinophilic cytoplasm,lightly stained.Under CLSM,by CK19 staining,the luminal structure in the middle of the cell mass was observed,while CK18 and CEA were expressed in this structure.Conclusion Under the induction of particular concentration of EGF,the in vitro cultured human ESCs seeded on the tissues engineering dermis could form into the tubular structure,which is similar to the in vivo sweat gland secretory cells in morphology and histology.

The training of professional postgraduates is one mode of medical postgraduate training,which more emphasizes the cultivation of clinical practice and ability.The problems existing in the training of professional degree postgraduates in dermatology are analyzed in this article,such as weak foundation of scientific research foundation,lack of supervision and evaluation,and the challenges from dermatology characteristics.The solutions including the training objectives,project design,the optimization of teaching process and evaluation,the training of comprehensive ability will be discussed in this article.The purpose is to improve the quality of training,to give some advice for providing high quality professional degree postgraduates.

Objective To investigate the influence of Wnt-3a signaling on aggregative growth of dermal papilla cells.Methods Recombinant adenovirus pAdEasy-1/Wnt-3a was constructed at first,and used to transfect cultured dermal papilla cells(DPCs).Then,the growth pattem of DPCs was observed by inverse microscopy;laser confocal microscopy and Western blot were utilized to detect the expressions of Wnt-3a,β-catenin and versican in low-passage DPCs,high-passage DPCs and transfected high-passage DPCs.Results The over-expression of Wnt-3a protein could effectively induce the aggregative growth of DPCs.Laser confocal microscopy showed that the fluorescence intensity of β-catenin and versican increased from 34.16±9.62 and 18.81±9.59 in high-passage DPCs to 87.35±17.28 and 92.18±18.39 in transfected high-passage DPCs(t=11.98,17.88,both P＜0.0 1),respectively.Also,Western blot proved a significant increase of expression intensity of β-catenin(15.27±2.17 vs 60.09±7.24,t=10.10,P＜0.01)and versican(19.32±2.46 vs 58.46±2.78,t=20.63,P＜0.01)in transfected high-passage DPCs compared with untransfected high-passage DPCS.ConclusionsWnt-3a signaling plays an important role in modulating aggregative growth of DPCs,which is mainly through the classical pathway with β-catenin as the key protein.As an important downstream gene of Wnt signaling,vesican is an indispensable part for the modulation of aggregative growth of DPCs.

Objective:To study the effect of seeding cells of tissue-engineered skins,keratinocytes and fibroblasts,on the phenotypes of langerhans cells (LC).Methods:Peripheral blood mononuclear cells were induced by cytokines,granulocyte macrophage-colony stimulating factor (GM-CSF),interleukin 4 (IL-4),transforming growth factor β1 (TGF-β1) to generate LCs,which were subsequently cocultured with allogenic keratinocytes or fibroblasts.Then the phenotypes of langerhans cells was detected by flow cytometer (FCM),and the degree of proliferation of autogeneic lymphocytes was assessed after stimulation with LCs.Results:Cultured LCs had low level expression of HLA-DR,CD80,CD86 and no expression of CD83.Cocultured with allogenic keratinocytes or fibroblasts,phenotypes of LCs did not change significantly.Simultaneously,LCs could not stimulate autogeneic lymphocytes proliferation.Conclusion:These data indicate that LCs cocultured with seeding cells of tissue-engineered skins remain immature state.It is suggested that seeding cells of tissue-engineered skins have low immunogenicity and are unable to induce significant immunological rejection of host after transplantation.

Objective To evaluate our experience in the diagnosis and surgical management of papillary thyroid microcarcinoma(PTMC). Methods Clinical data of 42 PTMC cases were retrospectively analyzed. Results Twenty-five clinically nonpalpable PTMC were detected by high resohition thyroid uhrasonography preoperatively.The diagnosis of PTMC was established intraoperatively by frozen biopsy in 14 out of 19 cases undergoing this procedure.Of the 42 patients,30 underwent a lobectomy,and in 11 out of 30 patients supplemented level Ⅵ lymph node dissection was performed.None of these patients had recurrence during follow-up.The recurrence in three patients with multffocal lesions and undergoing incomplete resection were observed at follow.up.The mulifocality of PTMC and ipsilateral residual volume of the thyroid were two predicting factors that significantly influence the postoperative recurrence(P＜0.05,P＜0.01 respectively)in patients with PTMC. Candus-ions PTMC is usually occult and eludes correct preoperative diagnosis.Most PTMC are clinically nonpalpable and may be detected by hish resolution thyroid ultrasonography and diagnosed by frozen section during the operation.Surgery is the most important treatment of PTMC.Lobectomy plus level Ⅵ lymph node dissection is the therapy of choice for PTMC patients at the stage of cN0.

Objective The microbiology specimen types were various and complex ,the system of main specimen types in micro-biological detection was established under the laboratory information system (LIS) for realizing the management target of quality control before microbiological analysis .Methods A total of 3304 submitted microbiological samples were collected from January 1 to 31 in 2015 .After setting the microbiological item application procedure of main specimen types in LIS ,1532 submitted microbio-logical specimens from June 20 to 24 were performed the statistics .The error rates of specimen types were compared before and af-ter setting .Then 1635 and 1340 submitted microbiological specimens were re-collected form July 9 to 13 and August 10 to 15 ;the change of error rates was continuously observed for comparing whether the statistical difference of error rates existing between be-fore and after setting .Results The error rate of submitted microbiological specimens before setting was 4 .6% (152/3304) ,which after setting was 1 .3% (20/1532)(χ2 =31 .224 ,P<0 .001) ,which during the continuous observation period maintained the lower level of 1 .04% (17/1635) ,χ2 =39 .658 ,P<0 .001) and 0 .9% (13/1340 ,χ2 =34 .673 ,P<0 .001) .Conclusion Re-setting the LIS reduces the error rate of microbiological specimen type ,effectively increase the working efficiency and reaches the quality control in-dex before microbiological analysis .

Objective To explore the effects of Shihusan on the intracellular free calcium of human retina cells and the protection from the damage of glutamate. To investigate its function and mechanism in retinal degeneration diseases, and to offer the basis on which to treat the similar disease clinically. Design Experimental study. Participants Retinal cells from normal human eye. Methods Cultivation of human retinal neural cells was perfonned. The changes of fluorescent density of intracellular free calcium of cultured cells labeled with Fluo3/AM before and after adding Shihusan and verapamil were detected with laser scanning confocal microscopy. In addition, the effect of Shihusan on calcium overload provoked by glutamic acid was observed. Main Outcome Measures The level of intracellular free calcium of retinal cells. Results The retinal cells adhered, spread and grew well in vitro. The fluorescence intensity increased 88% in glutamic acid group, decreased 14.8% and 57.3% in Shihusan group and verapamil group respectively. Shihusan couldn't decrease the intracellular Ca2+ that was increased by glutamic acid, but could inhibit the increasing tendency induced by glutamic acid. The difference was significant compared to control group (P=0.000). Conclusions Shihusan might decrease the level of intracellular free calcium in cultured human retinal cells, and resist the damage of glutamate. Its mechanism was related possibly to resisting calcium overload and inhibiting apoptosis.

Objective To investigate the clinical outcomes of surgical excision combined with recom binant interferon aipha-2b in the treatment of acral malignant melanoma (MM). Methods Fifteen patients with acral MM admitted to the department since 2004 were recruited into this study. The tumors varied from 1.8 mm to 3.9 mm in invasion depth. Thin tumors with an invasion depth of 1.8 - 2.0 mm were excised with a margin of lcm beyond the tumors, and those with an invasion depth of 2.0 - 3.9 mm were excised with a margin of 2 cm beyond the tumors. After excision, 4 cases of minor excision were sutured directly, 10 cases of large excision were repaired with adjacent skin grafts and flaps, 1 patient with the involvement of the first metatarsophalangeal joint underwent toe amputation followed by the repair of planta wound surface with the remaining skin on the dorsa of toes. Patients received intramuscular recombinant interferon alpha-2b for 3 months (3 million units daily for the first 3 days and 6 million units for the remaining days) following operation. Results There were 6 cases of MM in situ and 9 cases of invasive MM in this study. All the skin grafts and flaps survived. Within the 3-year follow up, relapse was observed only in 1 patient with invasive MM. Recovery was achieved in the functions of feet in all patients. Conclusion The excision of tumors with a margin determined by tumor thickness plus intramuscular interferon alpha-2h may improve the survival of patients with cutaneous MM in planta pedis with avoidance of amputation.

Objective To compare the epidermal shape built by different passages of keratinocytes and its ability of proliferation and differentiation in three-dimensional conditions.Methods Different passages of keratinocytes were used to construct tissue-engineered skin.The morphology of the tissue-engineered skin was observed with HE and PAS staining,while CK1/CK10,CK5/CK14,Ki67 were detected by immunohistochemical assays.Results All the tissue-engineered skin had a significant dermoepidermal structure.The stratification of 1st and 2nd passage skins were better,and 2nd passage epidermis was thicker than that in other passages (P＜0.05).Dermoepidermal structure in collagen type Ⅳ group binded more tightly,but collagen type Ⅳ had little effect on the thickness of the epidermis (P＞0.05).In collagen type Ⅳ group PAS stain was negative,indicating type Ⅳ collagen was unable to promote the reconstruction of BM in vitro.The Ki-67 proliferation index of the 2nd keratinocyte was similar to the normal skin,the remaining passages keratinocyte proliferation gradually decreased (P＜0.05) ; the 1st and 2nd passage skins expressed CK1/CK10 and CK5/CK14.Conclusions Keratinocytes before the 3rd passage have a better ability in the proliferation and differentiation,and so they are more suitable as seed cells for tissue-engineered skin.