Objective To evaluate the effects of tumor-associated macrophages on the proliferation,invasion and migration of human cutaneous malignant melanoma cells.Methods Cultured U937 human monocytic cells at logarithmic phase were classified into three groups to be pretreated with phorbol ester for 48 hours followed by 48-hour activation by phorbol ester (M polarization),lipopolysaccharide (LPS) at 25 mg/L (M1 polarization),and interleukin (IL)-4 at 15 μg/L (M2 polarization) respectively.Then,enzyme-linked immunosorbent assay (ELISA) was performed to determine the levels of IL-12p70 and IL-10 in the supernatant of these activated cells.A375 human malignant melanoma cells were divided into four groups to be cultured alone or with M-,M1-and M2-polarized macrophages respectively.After additional culture for different durations (24,48 and 72 hours),methyl thiazolyl tetrazolium (MTT) assay was conducted to estimate the proliferative activity,and Transwell assay to evaluate the invasion and migration activity,of the A375 cells.Results The proliferation of A375 cells was accelerated by coculture with M-and M2-polarized macrophages,but inhibited by that with M1-polarized macrophages,with significant differences among the four groups in the proliferative activity at 48 and 72 hours (all P ＜ 0.05),but not at 24 hours (P ＞ 0.05).Invasion assay showed that the number of A375 cells that migrated through Transwell chambers was significantly larger in M2 and M groups (147.00 ± 7.92 and 113.22 ± 8.15 respectively),but smaller in the M1 group (56.44 ± 7.55),than in the control group (84.11 ± 6.07,all P ＜ 0.05).Similarly,migration assay revealed a significant increase in the number of A375 cells that migrated through Transwell chambers in the M2 and M(p) groups (198.33 ± 8.22 and 156.00 ± 8.83 respectively),but a significant decrease in the M1 group (97.11 ± 6.75) as compared with the control group (123.89 ± 7.01,all P＜ 0.05).Conclusions The proliferation,invasion and migration of A375 cells can be accelerated by IL-4-activated M2-polarized macrophages,but decelerated by LPS-activated M1-polarized macrophages.Phorbol ester tends to induce monocytic cells to differentiate into M2-polarized macrophages.

Objective To estimate the effect of the enviromental pollutant 2,3,7,8-tetrachlorodibenzo-pdioxin (TCDD),a representative of the dioxin family,on the expression of cytochrome P4501A1 (CYP1A1) in cultured human immortalized SZ95 sebocytes in vitro,so as to improve understanding of the pathogenesis of chloracne.Methods SZ95 sebocytes were cultured with or without the presence of 10 nmol/L TCDD for two hours or three days.Real time fluorescence-based PCR was performed to quantify the mRNA expression of CYP1A1,immunohistochemistry and Western blot to determine the expression level of CYP1A1 protein,in the SZ95 cells.Chi-square test was done to compare the protein and mRNA expressions of CYP1A1 between untreated and treated SZ95 cells.Results Real time PCR showed that the mRNA expression of CYP1A1 was low in SZ95 sebocytes,and increased by 5.622 times after 2-hour treatment with TCDD(P ＜ 0.05).Immunohistochemistry revealed a weak expression of CYP1A1 protein in the cytoplasm and nuclei of untreated SZ95 sebocytes,which was also significantly enhanced by the TCDD treatment.Western blot results showed that the relative expression level of CYP1A1 protein was 4.233 ± 0.252 in SZ95 sebocytes treated by TCDD for three days,significantly higher than that in untreated sebocytes(0.123 ± 0.208,P ＜ 0.05).Conclusions There is a low expression of CYP1A1 mRNA and protein in SZ95 sebocytes,which can be upregulated by TCDD,suggesting that the CYP1A1 gene is a downstream target of the aryl hydrocarbon receptor responsible for the abnormal differentiation of human sebocytes.

Objective To analyze the changes of lymphocyte subsets in peripheral blood of patients with drug eruption . Methods 18 newly diagnosed patients were served as the drug eruption group ,and were subdivided into cephalosporin group (n=9) ,penicillin group(n=5) and Chinese medicine group(n=4) according to different sensitizing drugs .20 healthy people were taken as the control group .Flow cytometry were utilized to detect the percentages and absolute counts of T lymphocytes (CD3+ ,CD3+CD4+ and CD3+CD8+ ) ,B lymphocytes ,natural killer cell(NK) and natural killer T lymphocytes(NKT) in their peripheral blood . Results Differences of percentages of T lymphocytes (CD3+ ,CD3+ CD4+ ) ,B lymphocytes ,NKT cells between the drug eruption group and the control group showed statistical significant (P<0 .05) .Difference of percentages of CD3+ CD8+ lymphocytes of pa-tients between the drug eruption group and the control group demonstrated no statistical significant (P>0 .05) ,while that of abso-lute counts of T and B lymphocytes of patients was statistical significant between the drug eruption group and the control group (P<0 .05) .Conclusion The percentages of CD3+ ,CD3+CD4+ lymphocytes of patients with drug eruption decrease ,while those of NKT cells increase ,which may be related to the patients′immune regulation .

Objective To study the susceptibility of Neisseria gonorrhoeae to antibiotic agents from 1988 to 2002 in Shanghai. Methods The clinical isolates from patients with gonorrhea were collected and tested for their susceptibility to five antibiotics. Agarose-dilution-method was used to detect minimal inhibitory concentration (MIC) of anti-microbial agents including penicillin, tetracycline, spectinomycin, ciprofloxacin and ceftriaxone, and penicillinase producing Neisseria gonorrhoeae (PPNG) were tested with acidometric method. Results Susceptible strains to penicillin decreased from 11.28% in 1988 to 0 in 2002, MIC50 and MIC90 increased 8 and 4 times, respectively, the resistant rate and proportion of PPNG were 94.29% and 50.95%, respectively in 2002. The strains of high resistance to tetracycline increased from 0 in 1995 to 20.95% in 2002. The susceptible strains to ceftriaxone decreased from 100% in 1995 to 23.80% in 2002. The susceptibility to ciprofloxacin decreased significantly and resistant rate reached 99.05% in 2002. However, these strains were kept highly susceptible to spectionmycin. Concerning the multi-drug resistance, we found that the strains resistant to penicillin, ciprofloxacin and tetracycline simultaneously increased from 20.87% in 2001 to 23.30% in 2002, those resistant to both penicillin and ciprofloxacin reached to 70% in the past 2 years. Conclusions In Shanghai the resistant rates of Neisseria gonorrhoeae to antibiotics increased year by year in the past 15 years. The study indicates that spectinomycin and ceftriaxone should be the first choice for the treatment of gonorrhea at present and new sensitive antibiotic should be developed for the treatment of gonorrhea.