Objective To investigate the T cell clonality in patients with systemic lupus erythematosus (SLE) . Methods T cell clonality in peripheral blood lymphocytes from six patients with active SLE was analyzed using reverse transcriptase- polymerase chain reaction (RT- PCR) and GeneScan technique. Results T cell receptor (TCR) Vβ PCR products from two patients were derived from polyclonal T cells, however, TCR Vβ9, Vβ2, VβI, Vβ2 pioducts from the other four patients were derived from oligoclonal T cells respectively.ConclusionT cells from patients with SLE demonstrated a striking oligoclnality and their clonal expansion may be a specific response driven by autoantigens.

Objective To assess the value and clinical applicability of non-treponemal antigen serologic test and Treponema pallidum antigen serologic test in the diagnosis of congenital syphilis. Methods Totally,7800 newborns were collected from the out-patient and in-patient department. Toluidine red unheated serum test (TRUST) was conducted to quantitatively detect non-treponemal antibodies and Treponema pallidum particle agglutination assay (TPPA) to qualitatively detect specific treponemal antibodies in these subjects. The results of first serology test for syphilis were assessed for neonates with confirmed congenital syphilis. The sensitivity and specificity in the diagnosis of congenital syphilis were compared between TPPA and TRUST.Results Of the 7800 newborns undergoing syphilis serology testa, 60 were positive for TPPA and TRUST,including 15 cases of congenital syphilis, 47 were positive for TPPA but negative for TRUST, including 5 cases of congenital syphilis. The sensitivity and specificity were 75% (5/20) and 99.41% (7734/7780) respectively for TRUST in the diagnosis of congenital syphilis, 100% (20/20) and 98.88% (7693/7780) respectively for TPPA. Conclusion TPPA seems more applicable than TRUST to the screening for congenital syphilis.

Objective To evaluate the influence of chronic urticaria on the quality of life in patients by the DLQI, and to assess the factor structure and reliability of the DLQI. Methods The DLQI (Chinese version) was used to evaluate the quality of life of outpatients with CU. The reliability and factor structure of the DLQI were evaluated by reliability analysis and factor analysis. Results Eighty-five patients were asked to complete the DLQI and 13 patients were excluded for other complicated disorders or unanswered items in the DLQI. The mean age was 33.43 ± 0.90 years in the 72 patients with a median clinical course of 6 months. The DLQI score, which varied from 1 to 23 with an average of 9.46 ± 0.61, was uncorrelated to the patients' age,sex, or clinical course of CU (all P ＞ 0.05). The impact of CU was observed mainly on the "synptoms and feeling" (50%), followed by "work and study" (40.67%). Factor analysis indicated that there were two dimensions with eigenvalues greater than 1.0, accounting for 53.7% of the variances. Reliability analysis showed that the Cronbach's α value was 0.836, and reached 0.849 if the item 1 in the DLQI was deleted. An analysis of variance showed that the F value was 31.88 (P ＜ 0.01 ), and Hotelling T2 test yielded an F value of 29.87 (P ＜0.01 ). Conclusions CU has a moderate impact on the quality of life in patients. DLQI can be applied to the evaluation of quality of life in patients with CU with a high reliability, but more attention should be paid to the interference from item 1 to the reliability of the assessment.

Objective To investigate the association of HLA-A and -B alleles with syphilis in Shandong Han population. Methods The allele frequencies of HLA-A and -B were detected in 205 patients with syphilis and 5844 normal human controls by PCR-sequence specific oligonucleotide probe (PCR-SSOP)method. Results The patients with syphilis showed a higher frequency of HLA-A*02, B*15, B*40 alleles(all P＜0.01, Pc＜0.05) and a lower frequency of HLA-A*26 allele (P= 0.003, Pc = 0.039) than the normal human controls did. There was an increased frequency of HLA-B*15 and B*40 alleles in patients with symptomatic syphilis (both P＜0.01, Pc＜0.05), as well as an elevated frequency of HLA-A*02, 11, 29, B*15 and 40 alleles (all P＜0.01, Pc＜0.05) and a decreased frequency of HLA-A*30 and 33 in patients with asymptomatic syphilis(P=0.002, 0.026, Pc=0.001, 0.013 respectively), compared with the normal human controls. The frequency of HLA-A*30 allele was significantly higher in patients with symptomatic syphilis than in those with asymptomatic syphilis (P = 0.001, Pc = 0.013). Conclusions There seems to be an association between HLA-A*02, B* 15 and B*40 alleles and syphilis, between HLA-A*30 allele and symptomic syphilis, and between HLA-A*02, 11 and 29 alleles and asymptom1atic syphilis, in Shandong Han population.

Two patients were admitted to the hospital for 2-month history of pruritic eruptions on the forehead and 2-week history of pruritie eruptions on the leg, respectively. Both patients had a history of pet contact. Topical application of glucocorticoids did not work well. Dermatological examination revealed a patch measuring 5 cm ×6 cm on the forehead of one patient and a patch measuring 2 cm × 3 cm on the leg of the other patient. Both patches were surrounded by red papules and scaling. Microscopic examination of skin scales revealed hyphae and chain-like spores, and culture of skin scales grew Microsporum gyeum. Both the isolates of Microsporum gyeum were sensitive to ketoconazole, miconazole, bifonazole, terbinafine, and voriconazole. Both patients were healed after treatment with oral terbinafine and topical ciclopirox olamine.

Objective To explore the role of Toll like receptor 2 (TLR2) in the pathogenesis of acne vulgaris. Methods Venous blood was collected from 30 normal human controls and 90 patients with acne vulgaris. The patients were equally divided into three groups, i.e., mild, moderate and severe groups, according to disease severity. Flow cytometry was performed to detect the expression of TLR2 in PBMCs, and double antibody sandwich ELISA to measure the levels of serum interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α). Results A significant increase was observed in the expression of TLR2 in PBMCs, serum level of IL-8and TNF-α in the three groups of patients with acne vulgaris compared with the normal human controls (all P ＜0.01). The expression of TLR2 was positively correlated with the expression of IL-8 (r=0.382, 0.517,0.436,respectively, all P＜0.05) and TNF-α(r=0.641, 0.725, 0.593, respectively, all P＜0.05) in the patients with mild, moderate and severe acne vulgaris, respectively, and with the severity of acne vulgaris (r = 0.406,P＜0.01 ). Conclusion The expression level of TLR2 and related cytokines seems closely correlated with the severity of acne vulgaris.

Objective To establish a primary culture of human melanocytes from tiny skin sheets harvested by using a suction blister method, to carry out a serial subcultivation of the melanocytes with human amniotic membrane (AM) as a scaffold, and to observe the influence of AM on the adhesion, proliferation and dendrite development of melanocytes. Methods Tiny skin sheets were collected from the flexual forearm or lower abdomen of a healthy male volunteer by a suction blister method and melanocytes in the skin sheet were counted following Dopa staining under a microscope. The trypsinized skin sheets were scraped with a scalpel to harvest melanocytes which were subjected to a primary culture. Then, the melanocytes were inoculated onto fresh or cryopreserved AM followed by a culture for various durations (4, 8 and 12 days). The morphology and dendrite development of melanocytes were visualized under an inverted microscope after dopa-staining, cell viability evaluated by MTT assay, the adhesion to AM examined by hematoxylin and eosin (HE) staining protocol. Results The density of melanocytes was 1543.1±13.3 cells per mm2 and 857.4±101.7 cells per mm2 in skin sheets obtained from the forearm flexure and lower abdomen of the volunteer, respectively. A skin sheet of about 25.1 mm2 from approximately two blister roof was required to ensure the success of primary culture of melanocytes within 1 month. After culture on fresh or cryopreserved AM for 4, 8, and 12 days, most melanocytes were bi-polar with extended slender dendrites compared with those cultured in common cell culture medium. HE staining showed that melancytes adhered and were evenly distributed on the basement membrane of AM. MTT assay showed that the AM inhibited the proliferation of melanocytes, and no statistical difference was observed in the inhibitory effect between fresh AM and cryopreserved AM (P＞ 0.05). Conclusions Enriched with melanocyes, flexural forearm is a preferable donor site to offer skin sheets for primary culture of melanocytes. Human AM could improve the adhesive growth and dendrite development of melanocytes, and may serve as a promising bioscaffold for in vitro expansion of melanocytes.

Objective To construct tissue-engineered skin containing melanin with mixed culture of human keratinocytes (KCs) and melanocytes (MCs) on de-epidermized dermis (DED) in vitro. Methods Single-cell suspension was obtained by digestion of isolated preputial epidermis with pancreatin. Keratinocyte serum-free medium (K-SFM) and modified M254 culture medium were used to culture KCs and MCs respectively. Third-passage KCs were seeded into cell culture flasks and cultured for 48 hours; then, third-passage MCs were seeded into the same cell culture flasks with the MC/KC ratio being 1: 10 followed by a 5-day coculture. The suspension of third-passage KCs and MCs with the MC/KC ratio of 10:1 were seeded onto the surface of a prepared DED and maintained at the air-liquid interface for 11 days following a 4-day submerged culture.Subsequently, the constructed tissue-engineered skin was examined with HE staining, immunohistochemical staining for keratin and Masson-Fontana staining. Results After coculture in flasks for 5 days, KCs exhibited a typical paving-stone appearance, MCs with projected dendrites were scattered in the extracellular space between KCs. HE staining revealed 3 to 6 layers of cells with the formation of stratum corneum after mixed culture on DED for 15 days. Keratin protein was positive throughout the artificial epidermis, and melanin pigments were located in the basal layer of the epidermis as Masson-Fontana staining showed. Conclusions The co-culture of MCs and KCs can form single-cell layers with the contact between MCs and KCs in flasks, and construct tissue-engineered skin with melanin component on DED in vitro.

Objective To measure the expressions of HSP10 and 60 in cutaneous squamous cell carcinoma (SCC), basal cell carcinoma (BCC) and solar keratosis (AK) tissue. Methods Lesion samples were resected from patients with SCC (n = 50), BCC (n = 50) and AK (n = 50), and control samples were obtained from the normal skin adjacent to the operation sites of 14 of the 50 patients with SCC, BCC and AK. Immunohistochemical Envision two step method was used to detect the expression of HSP60 and 10 in the tissue samples.Results The expression of HSP10 was significantly higher in BCC tissue samples (Z = 3.24, P ＜ 0.001 ), but not in AK (Z= 0.74, P＞ 0.05) or SCC (Z= 0.52, P＞ 0.05) tissue samples than in the normal control tissue samples. Statistical significance was observed in the expression of HSP10 between AK and SCC and between AK and BCC tissue samples (both P ＜ 0.05), but not between SCC and BCC tissue samples (P ＞ 0.05 ). Elevated expression of HSP60 was found in AK, BCC and SCC tissue samples compared with the control samples (Z =-2.90, -2.15, -2.78,P ＜ 0.01, 0.05 and 0.01, respectively). Furthermore, the expression of HSP60 in SCC tissue samples was higher than that in BCC tissue samples (P ＜ 0.05 ) but similar to that in AK tissue samples. Conclusions There is likely to be a correlation between the high expression of HSP60 and biological behavior of SCC, and between the elevated HSP60 and HSP10 expressions and BCC initiation and development.

Objective To observe and compare the efficacy and safety of implantation of tissue expanders adjacent to or under keloid tissues for large keloids on anterior chest. Methods Between Mar 2006 and June 2009, a total of 17 patients with large keloid lesions on anterior chest received treatment with 21 tissue expanders,among which 12 were placed under the normal skin adjacent to keloids, and 9 were inserted under the keloid lesions. The scar size varied from 4.5 cm × 3.0 cm to 15.7 cm × 5.5 cm. The capacity was 70 to 400 ml for expanders adjacent to the keloid tissue, 80 to 500 mi for those beneath the keloid tissues. After tissue expansion for 6 to 8 weeks, the expander was removed and keloid lesions were resected followed by the repair of defect with expanded flaps. Further more, the patients received intraoperative local intradermal injection of betamethasone and postoperative superficial electron beam irradiation with divided doses of 7 Gy in 3 consecutive days within 1 week after the surgery. Follow-up varied from 12 to 50 months. Results Twenty expanders, except 1expander pocket which was removed ahead of time due to infection, were implanted successfully during the whole course of treatment. The main complication was expander exposure in 4 patients, including 1 expander adjacent to the keloids and 3 under keloid lesions, which showed no significant influence on secondary operation. Fifteen patients reported relief of symptoms and achieved satisfactory outcomes, while 2 patients, including 1 treated with expanders adjacent to the keloids and 1 with expanders under the keloid tissue, showed great suture tension and experienced delayed stitch removal followed by the recurrence of keloids after the operation.Conclusions The implantation of tissue expanders under the adjacent normal skin or keloid lesions is an ideal treatment option for large keloids on anterior chest. Regional suture tension is a direct contributor to the recurrence of keloid formation after surgical excision.