Objective:To explore the polarization of Th1 and Th2 immune response and the changes of transcription factor(T-bet and GATA-3) in BALB/c mice infected with toxoplasma gondii.Methods:Sixty BALB/c mice were infected with toxoplasma tachyzoite(1?104/m,0.5 ml) by intraperitoneal injection and thirty BALB/c mice only received an injection of same amount of normal saline as control.After injection,two mice of infection group and one mouse of control group were killed in odd number days.Then,the serum IFN-? and IL-4 were measured by ELISA,and the mRNA expression of T-bet and GATA-3 in mice spleens were detected by Real-Time RT-PCR.Results:In the infection group of mice,the IFN-? was remarkable increased at the fourth day and kept the peak from the 5th day to 7th day.Then,it decreased to normal level at 9th day postinjection;but IL-4 was remarkable increased at the 8th day and reached to the peak at 9th day.Then it decreased to normal level at 15th day postinjection.The mRNA of T-bet expression was increased at the third day and kept the peak from the 5th day to 7th day.Then,it decreased to normal level at 9th day postinjection.The mRNA of GATA-3 expression was increased at the 7th day and rised to the peak at 11th day.Then,it decreased to normal level at 13th day postinjection.There were not obvious changes of IFN-? and IL-4 or T-bet and GATA-3 in normal mice.Conclusion:The Th1 immune response is dominant in acute infection when mice are infected by toxoplasma.Then it biases toward Th2 response at 9th day to 13th day postinfection.The shift of Th1/Th2 polarization influences the finale of toxoplasma infection.

Objective:To study the gene expression spectrum of Th cell immune response in acute and chronic toxoplasma infection in BALB/c mice.Methods:BALB/c mice were infected with toxoplasma tachyzoite (1?104/m,0.5 ml) by intraperitoneal injection.Two mice were killed at 6th and 12th day postinjection.The immune response gene expressing levels in mice spleen cells were detected with 128 kind gene microarray chips.Partial of the results were determined with Real time-PCR.Results:There were 20 genes changed after toxoplasma infection.Eight of them (Gata-3,Ccr3,Ccr4,Bcl6,Nfatc1,CD80,Fos,CD69) were regulated upward and twelve genes (T-bet,CIITA,Irf1,Mapk9,Nfatc3,Fasl,Tyk2,Lat,Mapk10,Socs3,Socs6 and Yy1) were regulated downward.On the whole,expression of Th2 response genes presented a up trend,and Th1 response genes presented a down trend.The results of Real time-PCR were accordant with microarray in the expression of Gata-3 and T-bet gene.Conclusion:According to the Th cell immune response gene expression spectrum,Th1 immune response is dominant in the acute infention when mice are infected by toxoplasma.Then,it is biased toward Th2 immune response.

Objective:To study single nucleotide polymorphisms in the promoter region of TNF-? gene in Chinese Han,Miao and Buyi population in Guizhou province.Methods:The five polymorphism sites at-1031,-863,-857,-308,-238 in TNF-? gene promoter region were detected using PCR-RFLP and SSP-PCR.Results:The allele frequencies of TNF-? gene in promoter region in these three main ethons of population were observed.In Chinese Han population,the-238 G was 96% and-238 A was 4%.Inaddition,-308 G was 92% and-308 A was 8%,-857 C was 65% and-857 T was 35%,-863 C was 80% and-863 A was 20%,-1031 T was 48% and-1031 C was 52%;In Miao people,the frequency at the loci above were 15.0%,8.0%,26.0%,4.0% and 50.0%;And the frequency of these five loci above were 14.0%,6.0%,11.0%,16.0% and 50.0% in Buyi population.Conclusion:Single nucleotide polymorphisms of TNF-? gene exhibits some specific characters in Chinese Han and Miao and Buyi population in Guizhou province.

Objective To explore the effect of the superantigens staphylococcal enterotoxin A (SEA) and staphylococcal enterotoxin B (SEB) on the level and positive sero-conversion rate of anti-HBs antibody induced by HBsAg. Methods Sixty BABL/C mice were randomly divided into 5 groups. The mice in group 1 were administered 4?g recombinant HBsAg by subcutaneous injection in the groin once a week for three weeks. The manner of HBsAg administration in the other four groups was the same as that in group 1. One day after the first administration, the mice were administered 25?g SEA (group 2) or 25?g SEB (group 3) by subcutaneous injection in contralateral groin. In the second week after the first administration, the mice were administered 25?g SEA (group 4) or 25?g SEB (group 5) by subcutaneous injection in contralateral groin when HBsAg were given. Serum levels of anti-HBs antibody in the five groups were measured once a week until the end of experiment using ELISA. Results On the third week, the positive sero-conversion rate and serum level of anti-HBs in group 1 were 41.7% and 0.758?0.126, respectively. At the same time, the anti-HBs sero-conversion rate in the group 2 and group 4 was 100%, and there was significant difference in the anti-HBs level compared with group 1(P0.05). Conclusion SEA could remarkably elevate the level and positive sero-conversion rate of anti-HBs after injection of HBsAg, and significantly enhance the humoral immune response induced by HBsAg in mice.

The paper is to report the pharmacokinetics of sulfamethoxazole in healthy Han volunteers living at plain (PH) and native Han and Tibetan healthy volunteers living at high altitude (HNH and HNT). After healthy volunteers were administrated orally cotrimoxazole tablets, plasma concentration of sulfamethoxazole and metabolite N4-acetylsulfamethoxazole was determined by RP-HPLC, and plasma concentration-time data were analyzed by DAS 2.0 software to get the related pharmacokinetic parameters. The main pharmacokinetic parameters t(1/2) of sulfamethoxazole in PH, HNH and HNT were, respectively, 9.30 +/- 1.11, 10.99 +/- 1.23 and 10.44 +/- 1.05 h; tmax were 1.4 +/- 0.3, 2.0 +/- 1.1 and 1.8 +/- 0.4 h; Cmax were 94.42 +/- 15.26, 89.33 +/- 7.67 and 87.43 +/- 11.61 micro x mL(-1); AUC(0-t) were 1202.5 +/- 238.3, 1 434.7 +/- 193.9 and 1302.8 +/- 103.0 microg x h x mL(-1); AUC(0-infinity) were 1240.7 +/- 255.3, 1511.5 +/- 211.9 and 1363.9 +/- 116.5 microg x h x mL(-1); CL were 1.01 +/- 0.22, 0.81 +/- 0.12 and 0.89 +/- 0.08 L x h(-1) x kg(-1); V were 13.27 +/- 1.73, 12.81 +/- 2.15 and 13.28 +/- 1.20 L x kg(-1). Sulfamethoxazole pharmacokinetic parameters of HNH and HNT were significantly different from that of PH. The t(1/2) was significantly higher and the CL was significantly lower in HNH and HNT than that in PH, and the AUC(0-infinity) was significantly lower in HNT compared with HNH. This study found significant changes in the disposition of sulfamethoxazole under the special environment of high altitude hypoxia. This finding may provide some references for clinical rational application of sulfamethoxazole in HNH and HNT.

Objective To evaluate and monitor the efifcacy of GnRH analogs (GnRHa) therapy. Methods Thirty girls with central precocious puberty diagnosied by LHRH stimulation test were treated with GnRHa for 6-24 months. The LHRH stimula-tion test were performed again at 3 months after initiation of therapy and then every 6 months during treatment. The relationship of peark LH and clinical suppressing pubertal (including Turner stage, bone age, grwoth speed) were compared. The monitor effect of peak LH to efifcacy of GnRHa were eveluated. Results Ninety LHRH stimulation tests were performed, only 7 cases were found to have clinical pubertal development. After 6 months treatment, the base LH level of thirty girls (0.48±0.20) IU/L was signiifcantly lower than that before the treatment (0.75±0.35 IU/L) (P=0.000). The correlation coefifcient between base LH and peak LH was 0.62. The best correlation between clinical suppressing pubertal and LHRH stimulation test was achieved when peak LH was less than 2 IU/L (85.7%sensitivity, 100%speciifcity). Conclusions Base LH value can be used in preliminary as-sessment of the efifcacy of GnRHa therapy for girls with central precocious puberty. The peak LH less than 2 IU/L can be as the indicator of treatment efifcacy.

This study is to investigate the effect of high altitude hypoxia on the activity and protein expression of CYP2C9 and CYP2C19. Rats from plain (P) and rats with acute middle altitude hypoxia (AMH), chronic middle altitude hypoxia (CMH), acute high altitude hypoxia (AHH) and chronic high altitude hypoxia (CHH) were administered orally phenytoin sodium (PHT) and omeprazole (OMZ) to evaluate the activity of CYP2C9 and CYP2C19, separately. The serum concentrations of PHT and metabolite 4'-hydroxyphenytoin (HPPH) at 12 h after treatment and the serum concentrations of OMZ and metabolite 5-hydroxy omeprazole (5-OHOMZ) at 3 h after treatment were determined by RP-HPLC. The activity of CYP2C9 and CYP2C19 was evaluated by the ratio of HPPH to PHT and the ratio of 5-OHOMZ to OMZ, respectively. The protein expressions of CYP2C9 and CYP2C19 were determined by ELISA method. The activities of CYP2C9 (HPPH/PHT) in P, AMH, CMH, AHH and CHH were 0.67 +/- 0.31, 0.75 +/- 0.29, 0.76 +/- 0.23, 0.79 +/- 0.31 and 0.75 +/- 0.18, respectively, and the activities of CYP2C19 (5-OHOMZ/OMZ) in P, AMH, CMH, AHH and CHH were 0.17 +/- 0.06, 0.20 +/- 0.10, 0.11 +/- 0.05, 0.37 +/- 0.13 and 0.19 +/- 0.05, respectively. The protein expressions of CYP2C9 in P, AMH, CMH, AHH and CHH were 4.20 +/- 1.27, 3.95 +/- 0.81, 3.93 +/- 1.11, 4.32 +/- 1.03 and 4.12 +/- 0.86 ng x g(-1), respectively, and the protein expressions of CYP2C19 in P, AMH, CMH, AHH and CHH were 3.91 +/- 1.82, 3.63 +/- 2.07, 2.55 +/- 0.85, 4.78 +/- 2.37 and 3.51 +/- 1.03 ng x g(-1), respectively. The activities and protein expressions of CYP2C9 in AMH, CMH, AHH and CHH were not significantly different with those of P. The protein expressions of CYP2C19 in AMH, CMH, AHH and CHH were not significantly different with those of P, but the activity of CYP2C19 in AHH was significantly higher than that of P. This study found significant changes in the activity of CYP2C19 under the special environment of acute high altitude hypoxia.

Objective To investigate the effect of electron transfer system on the hyphal formation of Candida albicans. Methods Candida albicans was cultured in RPMI 1640 supplemented with 10% new-born calf serum in 5% CO2 at 37 ℃ with or without the presence of inhibitors or activators of electron transfer system. Growth curve, morphology and percent of filamentation were observed for Candida albicans. MTT assay was used to assess the viability of Candida albicans. Results The solvents (chloroform and dimethyl sulfoxide) had no significant effect on the growth of and filamentation in Candida albicans. After incubation with thenoyltrifluoroacetone (TTFA) or benzhydroxamic acid for 24 hours, yeast cells of Candida albicans predominated in the culture. The growth of Candida albicans was significantly inhibited in log phase by the incubation with classic respiratory chain inhibitors such as rotenone, antimycin A, oligomycin, sodium azide, TTFA and sodium malonate, compared with the controls (all P < 0.01). Benzhydroxamic acid, an inhibitor of alternative oxidative pathway, also significantly inhibited the growth of Candida albicans in log phase (t = 10.92, P < 0.01). After incubation with rotenone, antimycin A, oligomycin, sodium azide, TTFA, sodium malonate, benzhydroxamic acid and disodium gnanylate, the percentage of filamentation in Candida albicans at 12 hours was 87.49 ± 0.52, 48.75 ± 4.44, 50.33 ± 8.50, 99.00 ± 1.00, 1.60 ± 0.53, 94.01 ± 0.99, 0.00 ± 0.00 and 92.33 ± 2.08, respectively, and the growth of Candida albicans at 7 hours was inhibited by (1.34 ± 0.15)%, (70.61 ± 1.02)%, (50.63 ± 5.38)%, (17.80 ± 7.89)%, (45.17 ± 1.27)%, (10.75 ± 3.62)%, (72.46 ± 1.14)% and -(5.96 ± 4.07)%, respectively. Conclusions Hyphal formation of Candida albicans could be suppressed by inhibitors of classic respiratory chain or alternative oxidative pathway, and is mainly regulated by alternative oxidative pathway.

Objective To determine the extracellular enzymatic activity of common pathogens for onychomycosis, in the hope of finding virulence factors associated with the pathogenesis of onychomycosis. Methods Strains tested in this study included standard strains of common dermatophyte and non-dermatophyte fungi as well as clinical isolates of Trichophyton rubrum from patients with onychomycosis. All the tested strains were cultured in medium containing nail fragments at 25 ℃ for 10 to 21 days followed by the determination of the nail fragment-containing medium, a significant increase was observed in the activities of esterase, esterase lipase, N-acetyl-β-glucosaminidase and α-mannosidase in dermatophytes compared with non-dermatophytes (all P ＜ 0.05 ), as well as in the activity of N-acetyl-β-glucosaminidase in Trichophyton rubrum compared with the other tested species of fungi (all P ＜ 0.05). No significant difference was noted in the activity of extracellular enzymes, except for that of naphthol-AS-BI-phosphohydrolase, between the isolates of Trichophyton rubrum from patients with different ranges of scoring clinical index for onychomycosis (SCIO). Conclusions In specific conditions, the extracellular enzymatic activity of fungi isolated from patients with onychomycosis is associated with fungal species, and may have a certain influence on the manifestations of anychomycosis.

A 19-year-old man was admitted to the hospital for erythema and nodules on the face and postauricular region for 6 years. Microscopic examination of lesion scrapings revealed brown septate hyphae. A restricted, velvety and black colony grew on Sabouraud's dextrose agar. Slide culture on potato dextrose agar plate showed flask-shaped phialides at the apex of or around the hyphae with clear collarettes and flaring apex,mucilage-encapsuled, round to oval, semi-endogenous phialosporae accumulating at the apex of the phialides,giving a flower-like appearance. Anti-fungal susceptibility test showed that the fungus was sensitive to itraconazole, terbinafine and amphotericin B, but resistant to fluconazole. Sequence analysis of the ITS1-ITS4 region revealed a 98% consistency with the reference sequence of ITS1-ITS4 of Phialophora verrucosa. On the basis of above findings, the patient was diagnosed with cutaneous phaeohyphomycosis. Clinical improvement was seen after treatment with oral itraconazole (400 mg/d).