Objective To study the effects of bFGF on the proliferation of cultured human melanocytes, and to seek a quick method for in vitro culture of human melanocytes. Methods Melanocytes were isolated from human foreskin, and divided into two parts to be cultured with or without the presence of bFGF (0.3 μg/L). Second-passage melanocytes were identified with immunochemical stain. The growth of melanocytes was observed every 3 days for 12 days. Third-passage melanocytes were treated with various concentrations (0.3 - 2.1 μg/L) of bFGF for 72 hours followed by the detection of proliferation of and trosinase activity in melanocytes. Results Human melanocytes were obtained from primary culture in medium containing certain concentrations of bFGF, which were identified with immunohistochemical stain. The morphology of cultured melanocytes varied with growth stage of cells. The bFGF-treated melanocytes appeared to grow more rapidly than untreated melanocytes. Further more, a significant increase was observed in the proliferation rate of melanocytes treated with bFGF of 0.3 and 0.6 μg/L (P＜0.05 or 0.01 ) and tyrosinase activity in melanocytes treated with bFGF of 1.5 and 1.8 μg/L (P ＜ 0.05 or 0.01 ) in comparison with the untreated melanocytes.Conclusions The addition of certain concentrations of bFGF to defined medium can benefit the primary culture of melanocytes and make it possible to get large quantities of purified melanocytes with high viability in short periods. Certain concentrations of bFGF can up-regulate the proliferation of and tyrosinase activity in melanocytes.

Oncomelania hupensis which plays an important role in the transmission of schistosomiasis japonica is the only intermediate host of Schistosoma japonicum and carrying out studies on its subspecies differentiation and genetic variation will have an important significance for schistosomiasis control. In this paper the research progress of taxonomy and discrimination techniques of O. hupensis is reviewed at four levels namely the morphological level the cellular level the protein level and the DNA molecule level and DNA sequencing technology is considered as the ideal taxonomy and discrimination technique of O. hupensis and the future research emphasis.

Objective One of the characteristics of rheumatoid arthritis was multiple joints involved symmetrically. The purpose was to study the method of early diagnosis and treatment for the atypical rheumatoid arthritis involving solitary joint. Methods From April 1997 to December 2002, 5 cases which were 1 male and 4 female patients with an average age of 36.8 years (range, 23 to 47 years), suffered from the swelling and pain of knee joint, and were diagnosed as early rheumatoid arthritis. The interval of the onset to surgical treatment ranged from 2 months to 3 years with a mean of 16.2 months. The patients were evaluated with serum test, joint fluids examination and radiological imaging. They were treated by the synovectomy under the arthroscopy, while the appearances of the joint under arthroscopy were recorded, and the synovial tissue was sent to the pathological examination. The improvements of the joints symptoms and the range of motion were assessed at final follow up. Results During the operation, the light green and little muddy joint fluids were found in all patients. ESR of the patients ranged from 29 to 51 mm/1h with an average of 32.2 mm/1h, all of which were higher than normal value, whereas, 3 cases had positive rheumatoid factor and 4 cases showed significant higher ?2-MG. The radiographic findings were osteoporosis in 3 cases, soft tissue swelling in 4 cases and narrowed joint space in 1 case. The synovium were proliferated and enlarged in dark violet, there were creeping pannus and necrosis fibrin deposition in the cartilage. All of the patients were diagnosed as rheumatoid arthritis confirmed by pathological changes of the synovium. The patients were followed up from 4 months to 5 years (mean, 18.7 months). Pain released in 5 cases, swelling reduced in 3 cases, the range of motion improved by 19 degrees. Unfortunately, all of the patients developed later into typical rheumatoid arthritis after the mean interval of 10.9 months (range, 3 to 25 months), the symptoms were consistent with the criteria of American Rheumatoid Association . Conclusion Solitary joint with rheumatoid arthritis is rare and may be the early status of rheumatoid arthritis. The specific appearance under the arthroscopy prompt the early diagnosis of the knee rheumatoid arthritis, and synovectomy by arthroscopy may be benificial to release the symptoms of rheumatoid arthritis.

ObjectiveTo observe the protective effect of high-dosage methylprednisolone (MP) after lumbar spinal stenosis operation. Methods20 Patients with lumbar spinal stenosis accepted laminectomy. The treatment group (11 cases) received MP 30 mg/kg when the skin incised, and followed with MP 40 mg twice a day for 3 d after operation. The control group (9 cases) received MP 40 mg twice a day for 3 d after operation. They were assessed with JOA scale before and after operation. ResultsThe JOA score was (9.25±2.12) and (9.53±2.10) in treatment group and control group before operation (P>0.05), and was (13.43±2.01) and (11.21±2.13) 1 week after operation (P<0.05), (14.62±2.15) and (13.04±2.11) 3 months after operation (P<0.05). The neurological symptom deteriorated in 1 cases of control group. No peptic ulcer, poor wound healing or other associated complications has been found. ConclusionHigh-dosage methylprednisolone may protect the spine cord intraoperative the lumbar spinal stenosis.

Objective To investigate the results of different measuring procedures of hemoglobin A1c (HbA1c) trueness verification scheme in China.Methods Cross sectional survey.The data were collected via the External Quality Assessment (EQA) software from laboratories participated in the First HbA1c trueness verification EQA.Then the collected data were divided into several groups based on laboratory instruments and the data from less than 5 group were excluded.The observed imprecision, bias and sigma (σ) were calculated and the bias％ and CV％ were drew in the sigma chart.The average bias％, CV％ and weighted average σ of each level were also calculated.Results Total 123 laboratories were divided into 9 groups and setting 6％ as the Allowable Total Error, the average bias％, CV％ and weighted average σ of 201411 (target value was 5.4％) were 3.70％, 4.55％ and 0.51 respectively σ, of 201412 (target value was 7.8％)were 2.42％ , 3.56％ and 1.24σ respectively.None of the group achieved the 2σ quality of 201411, and 1 group achieved the 2σ quality of 201412.Conclusions There are obvious biases among the results of many measuring systems and the target value assigned by reference measuring procedures of HbA1c, as well as the imprecision.The Sigma External Quality Assessment Chart is a visual tool, indicating that the quality of measuring systems necessitate improvement therefore to ensure the reliability of results and make better use of HbA1c in clinical application.

Key factors for interpreting proficiency testing (PT)/external quality assessment (EQA) results are knowledge of the commutability of the samples used and the process used for target value assignment.A commutable PT/EQA sample,which would be seen as patients' samples,has the agreement of results among different measurement procedures and demonstrates the same numeric relationship.The value assignment for the sample can be done by using of a reference measurement procedure.Its results can be used to assess the accuracy against a reference measurement procedure or a designated comparison method.Noncommutable PT/EQA samples frequently have a matrix-related bias of unknown magnitude,and can't give meaningful information about the relationship of results for patients' samples among different measurement procedures.Its results must be compared to the peer group mean or the median of results from participants who use measurement procedures that are expected to have the same or very similar matrixrelated bias.

Objective To investigate the phenotype,number and distribution of inflammatory cells in early and late stages of spontaneous regression of halo nevi,and to elucidate the immunological mechanisms for spontaneous regression of these nevi.Methods Halo nevi,their surrounding non-lesional skin,and normal control skin were examined by immunohistochemical staining with monoclonal antibodies to CD3,CD4,CD8,CD20,CD1a,CD56 and CD68.Staining results were observed and analyzed by the computer image analysis system,image-pro plus 6.0.Results The number of CD4+,CD8+,CD20+,CD1a+cells,along with the diameter of CD1a+and CD68+ cells was significantly increased in the lesions of early and late stage of spontaneous regression of halo nevi than in non-lesional skin and normal control skin(both P＜0.01).The ratio of CD8+/CD4+ cells in the lesions of late stage of spontaneous regression was also higher than that in the lesions of early stage (2.05∶1 VS 1.82∶1).A massive infiltrate of CD8+ cells was observed in the nests of nevus cells.ConclusionsCD4,CD8,CD20,CD1a,CD56 and CD68 positive cells are all involved in the spontaneous regression of halo nevi,and CD8+ cells may play a predominant role in this process.

BACKGROUND: Posterior spinal fusion is a process of bone fusion under special anatomical and biological effects, which affects by many factors. With the development of bone tissue engineering, in vitro constructed tissue engineered bioactive periosteum provides a new approach for solving this problem. OBJECTIVE: To evaluate the effect of in vitro constructed tissue engineered bioactive periosteum in treating lumbar intertransverse process fusion in rabbits. METHODS: In vitro constructed tissue engineered bioactive periosteum was implanted into lumbar intertransverse process of 24 healthy adult New Zealand rabbits. Three different materials were implanted into 3 transverse process gaps (Left L_(4,5,6), Right L_(4,5,6) of each animal. Namely, bone marrow mesenchymal stem cells (BMSCs) combined pig small intestine submucosa (SIS) were implanted into the right L_(4,5) of rabbits in the composite scaffold group; pure SIS was implanted into the right L_(5,6) of rabbits in the pure scaffold group; and autogeneic ilium was implanted into the left L_(5,6) of rabbits in the autogeneic ilium group. All rabbits were sacrificed at 12 weeks after operation to perform gross, imaging and histological observation. RESULTS AND CONCLUSION: The gross observation showed that there had no significant difference between the composite scaffold and autogeneic ilium groups, but the difference was significant compared with the pure scaffold group. lmaging observation showed that the trabeculae was formed in lumbar intertransverse of rabbits in the composite scaffold and autogeneic ilium groups, however, no bone density could be seen in the pure scaffold group. Type I collagen and osteocalcin were strong positive expressed in the composite scaffold group, which had obvious difference to the autogeneic ilium group. No positive expression could be found in the pure scaffold group. It suggested that tissue engineered bioactive periosteum constructed by BMSCs combined with SIS is a well alternative to autogenous graft materials for spinal fusion.

Objective To study the in vitro culture condition for melanoblasts from human foreskin tissue. Methods The skin tissue taken from foreskin of children was treated with 0.5% dispase Ⅱ to separate epidermis from dermis, then with trypsin to obtain single cell suspension, which was cultured in modified medium for melanoblasts, i.e., MCDB254 medium supplied with several cell growth factors. Finally, melanoblasts were obtained based on the difference of adhesion speed. The morphology and proliferation of cultured melanoblasts were observed under a light microscope. DOPA staining, immunostaining with anti- S-100 and -tyrosinase related protein 2 (TRP2) antibodies, and transmission electron microscopy were per- formed to identify the cultured melanoblasts. Results The cultured human melanocytes displayed a match-like shape, scattered arrangement, syrmnetric double poles, slim cell body, highly refractive nuclei; meanwhile, the melanoblasts exhibited plentiful cytoplasm, large volume, bipolar or irregular shape and clonal growth. Additionally, the melanocytes were positive for TRP2, S-100 and Dopa staining, while the melanoblasts were positive only for TRP2. Electron microscopy revealed the presence of mature melanin granules (stage Ⅲ-Ⅳ ) in melanocytes but immature melanin granules (stage Ⅰ ) in melanoblasts. Conclu- sion Stable pure culture of melanoblasts has been realized with the reformed medium, which may lay a foundation for the investigation into the mechanism of epidermal pigmentation.