Ecological surveys of the Cryptococcus species complex in China.

BACKGROUND: Despite recent reports on the molecular epidemiology of cryptococcal infections in China, clinical isolates have been mostly reported from human immunodeficiency virus (HIV)-negative patients, and environmental isolates from China have rarely been included. The aim of this study was to investigate the ecological profile of Cryptococcus (C.) neoformans and C. gattii in China. METHODS: A survey was performed in 10 cities from 20°N (North latitude) to 50°N and in a Eucalyptus (E.) camaldulensis forestry farm at the Guixi forestry center, China. RESULTS: Six hundred and twenty samples of pigeon droppings from 10 cities and 819 E. camaldulensis tree samples were collected and inoculated on caffeic acid cornmeal agar (CACA). The brown-colored colonies were recultured to observe their morphology, growth on canavanine-glycine-bromothymol-blue (CGB) medium, phenol oxidase and urease activities, serotype and mating type. There were obvious differences in the positive sample rates of C. neoformans in pigeon droppings collected from the different cities, ranging from 50% in the cities located at latitudes from 30°N - 40°N, 29% at 20°N - 30°N and 13% at 40°N - 50°N. CONCLUSIONS: There were no differences in positive bevy rates (approximately 80%) among the three grouped cities. Mycological tests of 101 isolates purified from pigeon droppings revealed that they were C. neoformans var. grubii. We also observed variable capsular size around the C. neoformans cells in colonies with variable melanin production and the bio-adhesion of the natural C. neoformans cells with other microorganisms. One urease-negative C. neoformans isolate was isolated from pigeon droppings in Jinan city. No C. gattii was isolated in this study.

mRNA expression of chemokine receptors on peripheral blood mononuclear cells and correlation with clinical features in systemic lupus erythematosus patients.

OBJECTIVE: To investigate the expressions of chemokine receptors and interleukin (IL) receptors on the peripheral blood mononuclear cells (PBMCs) from systemic lupus erythematosus (SLE) patients and their correlations with clinical features as well as SLE disease activity index (SLEDAI). METHODS: The mRNA expressions of chemokine receptors and IL receptors on PBMCs of 93 SLE patients and 30 healthy controls were detected by reverse transcription-polymerase chain reaction, including CCR2, CCR3, CCR4, CCR5, CCR6, CCR8, CXCR3, CXCRS, CX3CR1, XCR1, IL-4R, and IL-10R. The clinical features of SLE patients were recorded. The correlations of chemokine receptors and IL receptors mRNA expressions with clinical features as well as SLEDAI were assayed using linear regression analysis. RESULTS: The level of CCR5 mRNA in SLE patients (including active and inactive SLE) was significantly higher than that in healthy controls (P < 0.05), and there was no significant difference between active and inactive patients in this respect (P > 0.05). CX3CR1 mRNA expression significantly increased from healthy control to inactive SLE to active SLE in sequence. The others (except for CCR8, CXCR3, and IL-10R) in active SLE patients were significantly higher than those in both inactive SLE patients and healthy controls (all P < 0.05). There were positive correlations between SLEDAI and CCR2 (r = 0.424, t = 4.313, P < 0.001), CCR3 (r = 0.518, t = 5.410, P < 0.001), CCR4 (r = 0.376, t = 3.851, P < 0.001), CCR6 (r = 0.457, t = 4.513, P < 0.001), CXCR5 (r = 0.455, t = 4.629, P < 0.001), CX3CR1 (r = 0.445, t = 4.523, P < 0.001), as well as XCR1 (r = 0.540, t = 5.445, P < 0.001). And CCR5 mRNA expression level was positively correlated with IL-4R mRNA (r = 0.313, t = 2.353, P < 0.05). The patients with myositis and cutaneous vasculitis simultaneously showed lower levels of CCR5 and CX3CR1, and CCR5 expression was negatively correlated with the scores of SLEDAI in SLE cases accompanied by photosensitivity (r = 0.426, t = -2.155, P < 0.05). CONCLUSION: Increased expressions of CCR5 and CX3CR1 on PBMCs may be indicators in clinical survey for SLE.

[The methylation locus and frequency pattern on p16 INK4a gene promoter CpG in epidermis of patients with psoriasis].

OBJECTIVE: To investigate the CpG methylation locus and frequency pattern on p16 INK4a gene promoter in epidermis of p16 INK4a methylated patients with psoriasis vulgaris. METHODS: The DNA specimens were obtained from epidermal lesion of 50 plaque psoriatic patients. Methylation specific PCR and DNA sequencing were used to detect the frequency and locus of methylation in p16 INK4a gene promoter region. RESULTS: Approximately 50% CpG was methylated in p16 INK4a methylated patients, methylation was found in specifical locus of p16 INK4a gene promoter. CONCLUSION: The distinct methylation pattern is showed on the p16 INK4a gene promoter region in patients with psoriasis.

Correlation between some Th1 and Th2 cytokine receptor gene polymorphisms and systemic lupus erythematosus in Chinese patients.

BACKGROUND: Genetic factors seem to play a significant role in the susceptibility to systemic lupus erythematosus (SLE). Some researchers have described amino acid polymorphisms within the interleukin-4 receptor (IL-4R) [isoleucine50valine (Ile50Val), arginine576glutamine (Arg576Gln), etc.], the IL-10R (G241A and G520A), and the interferon-gamma receptor (IFN-gammaR) [valine14methionine (Val14Met) and glutamine64arginine (Gln64Arg)]. METHODS: The Ile50Val genotypes were examined by the reverse transcription-polymerase chain reaction-single-strand conformation polymorphism (RT-PCR-SSCP) method and DNA sequencing. The RT-PCR-restriction fragment length polymorphism (RT-PCR-RFLP) method was used to detect the Arg576Gln, G241A, G520A, Val14Met, and Gln64Arg genotypes. RESULTS: There were significant differences in the genotype frequencies of Ile50/Ile50, G520/G520, and G520/A520 between the SLE group and healthy control group. There was a positive correlation between the IL-4R Ile50/Ile50 genotype and the susceptibility to SLE (P = 0.022). This was also found for the IL-10R G520/G520 (P = 0.004) and G520/A520 (P = 0.055) genotypes. The risk of SLE susceptibility was increased in individuals with the genotype combinations Ile50/Ile50 and Gln576/Arg576 (P = 0.056) and G241/G241 and G520/G520 (P = 0.004). The IFN-gammaR2 Arg64/Arg64 genotype decreased the risk of SLE (P = 0.047). A decreased risk of development of SLE was detected in individuals with the genotype combination IFN-gammaR2 Arg64/Arg64 and IFN-gammaR1 Val14/Val14 (P = 0.004). CONCLUSIONS: The IL-4R Ile50/Ile50 and IL-10R2 G520/G520 and G520/A520 genotypes were shown to determine the susceptibility to SLE in a Chinese population, as were the combinations Ile50/Ile50 and Gln576/Arg576, and G241/G241 and G520/G520.

The interferon-gamma receptor gene polymorphisms (Val14Met and Gln64Arg) are not associated with systemic lupus erythematosus in Chinese patients.

Genetic polymorphism is a difference in DNA sequence among individuals, groups, or populations that give rise to different forms. Differences in DNA sequences that occur naturally in a population. Single nucleotide substitutions, insertions and deletions of nucleotides and repetitive sequences (microsatellites) are all examples of polymorphism. The position at which such a sequence difference is found is a polymorphic site. A single nucleotide substitution is called a single nucleotide polymorphism (SNP). SNPs can occur in coding parts of the gene. If they result in genetic code change, amino-acid polymorphism would occur. The heterodimeric IFN-gamma receptor (IFNGR) complex was made up of two receptor subunits including IFNGR-1 and IFNGR-2. There exist five dbSNP alleles in IFNGR1 exon region and six dbSNP alleles in IFNGR2. Some researchers had found that the greatest risk of the development of systemic lupus erythematosus (SLE) be detected in the individuals who had the Met14/Val14 genotype or the combination of IFNGR1 Met14/Val14 genotype and IFNGR2 Gln64/Gln64 genotype in Japanese patients. So we aimed to assess the association between two polymorphisms within the IFNGR gene (A88G and A839G) and SLE in Chinese patients. This study included 154 patients with SLE and 159 unrelated healthy controls. We examined the IFNGR genotype by the reverse transcription-polymerase chain reaction (RT-PCR)-single-strand conformation polymorphism method, RT-PCR-restriction fragment length polymorphism method and DNA sequencing. Genotype frequencies between SLE patients and controls were compared and relationship between genotype frequencies and clinical manifestations of SLE were evaluated. We found that IFNGR2 Arg64/Arg64 genotype decrease the risk of SLE (OR = 2.326, 95% CI 1.181-4.581, Fisher P = 0.015), and the same as IFNGR2 Arg64/Arg64 genotype and IFNGR1 Val14/Val14 genotype combination (OR = 2.420, 95% CI 1.206-4.854, Fisher P = 0.013). The allelic frequency of Val14/Met14 is significantly higher in the patients with oral ulcer or thrombocytopenia when compared with patient without these clinical feature (OR = 4.630, 95% CI 1.370-15.640, Fisher P = 0.021; or OR = 6.368, 95% CI 2.009-20.191, Fisher P = 0.003). On the contrary, the allelic frequency of Val14/Val14 is lower in the patients with oral ulcer or thrombocytopenia than those without these clinical feature (OR = 0.216, 95% CI 0.064-0.730, Fisher P = 0.021; or OR = 0.157, 95% CI 0.050-0.498, Fisher P = 0.003). And after data analysis, we also find that the allelic frequency of Gln64/Gln64 is lower in the patients with arthritis when compared with patient without arthritis (OR = 0.369, 95% CI 0.166-0.818, Fisher P = 0.017). We can conclude that the IFNGR polymorphisms (Val14Met and Gln64Arg) are protective in SLE in Chinese patients. We describe a novel association between Val14/Met14 carriage and patients with oral ulcer or thrombocytopenia.

[Phenomena and pathological significances of the methylated p16 promotor in DNA derived from plasma and blood cells of patients with systemic lupus erythematosus].

OBJECTIVE: To detect the methylation status of p16 gene promotor in DNA derived from plasma and blood cells of patients with systemic lupus erythematosus (SLE) , and it's relationship with clinical symptoms. METHODS: p16 promotor methylation in plasma and peripheral blood cells (PBCs) DNA were simultaneously detected with the methylation specific PCR (MSP) method in 24 active SLE patients, 21 inactive SLE patients, as well as 20 healthy controls. RESULTS: In the plasma DNA, p16 gene methylation ratio (MP%) was higher in SLE patients than in the healthy controls (64.4% vs. 5.0%, P < 0.05). MP% in the active SLE patients was significantly higher than that in the inactive SLE patients (83.3% vs. 42.9%, P < 0.05). In the PBCs, p16 gene methylation ratio (MC%) in the healthy controls was significantly higher than that in SLE (80.0% vs. 48.9%, P < 0.05). MC% in the active SLE patients (29.2%) was the lowest among three groups. There was no significant difference between the inactive SLE patients and healthy controls (71.4% vs. 80.0%, P > 0.05). Each patient could be judged as one of the four methylation patterns: MP/MC, UP/MC (UP: unmethylated plasma p16) , MP/UC (UC: unmethylated PBCs p16) , and UP/UC. The ratios of MP/ MC and UP/UC were similar between the active and inactive SLE patients. However, different distributions of other two patterns were found in the active and inactive SLE patients as UP/MC 4.2% vs. 42.9% (P <0.05) and MP/UC 58.3% vs. 14.3% (P < 0.05), respectively. The active SLE patients with MP/UC and the inactive SLE patients with UP/MC showed different clinical symptoms and laboratory examinations. Significant correlation was found between the disease activity index for lupus patients (SLEDAI) scores and MP% (r = 0.93), between the SLEDAI scores and MC% (r = - 0.96) also between MC% and MP% (r = - 0.79). CONCLUSION: The p16 methylation assay provides available information for the diagnosis, judgment of disease activity, as well as novel insights into the pathogenesis underlying this disease.

[Itraconazole in the treatment of superficial candidal infections: twelve years' clinical experience].

Itraconazole has been used to treat superficial candidal infections in China for 12 years with promising efficacy and safety. This article retrospectively reviewed literatures published in the mainstream journals in China with an attempt to find a reasonable therapy for Chinese populations.

Fine mapping of a semi-dwarf gene brachytic 1 in barley.

RFLP markers isolated from barley, wheat and rice were applied to construct a fine structure map of brachytic1, a semi-dwarf gene located on chromosome 1(7H) short arm in barley. The map covered 15.2 cM with the average distance 0.8 cM between markers. A barley cDNA clone, MWG2074B co-segregated with brh1 gene in the test population. Another major band of this clone MWG2074A was 0.8 cM away from brh1 toward centromere. CDO545 and BCD129 were two flanking markers mapped on both sides of brh1, toward distal and pistal, respectively. CDO545 fitted the systenic region of rice genome, chromosome 6 short arm perfectly. However, two major bands of MWG2074 could not be mapped to the target position of rice genome.