Tu Yating

A study on the expression of interleukin(IL)-10 and IL-12 P35, P40 mRNA in the psoriatic lesions

SummaryTo investigate the possible role of interleukin (IL)-10 and IL-12 in the pathogenesis of psoriatic lesions and to supply theoretical basis for the gene therapy for psoriasis, the expression of IL-10 and IL-12 P35, P40 mRNA in 12 cases of psoriatic lesions and 6 normal skin tissues was detected by using RT-PCR technique. The results showed that the expression of IL-10 mRNA in the psoriatic lesions was significantly lower than that in the normal skin tissues (P<0. 001). The expression of IL-12 P35 was positive both in the psoriatic lesions and in the normal skin tissues. IL-12 P40 mRNA was expressed positively only in the psoriatic lesions but negatively in the normal skin tissues. It was suggested that IL-12 might take an important role in the occurrence and progression of psoriasis, but IL-10 might have certain role in the regression of psoriasis.

Susceptibility of mixed infection ofUreaplasma urealyticum andMycoplasma hominis to seven antimicrobial agents and comparison with that ofUreaplasma urealyticum infection

In order to investigate the susceptibility of mixed infection of Ureaplasma Urealyticum (UU) and Mycoplasma Hominis (MH) to 7 kinds of antimicrobial agents and comparison with that of UU infection in NGU patients, the in vitro susceptibility was determined by using microdilution method. The positive results were analyzed. The results showed that the sequence of susceptibility to 7 kinds of antimicrobial agents for both UU infection group and UU-MH mixed infection group was almost the same from the highest susceptibility to the lowest accordingly: Josamycin, Doxycycline, Minocycline, Sparfloxacin, Roxithromycin, Ofloxacin and Azithromycin. The total drug resistance rate for UU-MH mixed infection group (97.67%) was significantly higher than that for UU infection group (44.67%, P < 0.01). The highest drug resistance rate in UU group and UU-MH mixed infection group was 31.33% (Ofloxacin) and 90.48% (Azithromycin) respectively. UU-MH mixed infection showed an increased drug resistance and changes of drug resistance spectrum.SummaryIn order to investigate the susceptibility of mixed infection ofUreaplasma Urealyticum (UU) andMycoplasma Hominis (MH) to 7 kinds of antimicrobial agents and comparison with that of UU infection in NGU patients, thein vitro susceptibility was determined by using microdilution method. The positive results were analyzed. The results showed that the sequence of susceptibility to 7 kinds of antimicrobial agents for both UU infection group and UU-MH mixed infection group was almost the same from the highest susceptibility to the lowest accordingly: Josamycin, Doxycycline, Minocycline, Sparfloxacin, Roxithromycin, Ofloxacin and Azithromycin. The total drug resistance rate for UU-MH mixed infection group (97.67%) was significantly higher than that for UU infection group (44.67%,P<0.01). The highest drug resistance rate in UU group and UU-MH mixed infection group was 31.33% (Ofloxacin) and 90.48% (Azithromycin) respectively. UU-MH mixed infection showed an increased drug resistance and changes of drug resistance spectrum.

Two case report studies of Langerhans cell histiocytosis with an analysis of 918 patients of Langerhans cell histiocytosis in literatures published in China

Langerhans Cell Histiocytosis (LCH), which was previously designated histiocytosis X, is a group of diseases characterized as hyperplasia and dissemination of Langerhans cells. LCH may involve one or many body systems or tissues, such as skin, lymph nodes, bone, bone marrow, liver, spleen, lung, thymus, gastrointestine, endocrine gland, mouth, ears, and central nervous system. The clinical manifestation of this group of diseases varies from single organ to multiple organs, from newborn to the old, and is especially highly prevalent among children. LCH has been recognized as a rare disease in the past, whereas the incidence has significantly increased in recent years with the progress in diagnostic techniques. To summarize the feature of skin lesions in LCH, we present a retrospective clinical analysis of two patients with LCH diagnosed in our hospital and another 916 cases reported in China over the past 10 years. Clinical data

Comparison of the effects of three different anti-fungus drugs on Candida albicans of murine vaginal mucosa

To compare the therapeutic effects of three different anti-fungal drugs (i.e., terbinafine, fluconazole and intraconazole) in the treatment of experimental vaginitis caused by Candida albicans (C. albicans) in mice, the fungal vaginitis model was established in female ICR mice by intravaginal inoculation of suspension of C. albicans after the animal had been pretreated with estradiol. Mice were divided at random into different groups and then respectively treated with terbinafine, fluconazole and intraconazole given by gastrogavage. The burden of the fungus in the vaginal lavage fluids in the mice of the different groups was measured dynamically at different time points after the beginning of the drug treatment. The fungal burdens in the vaginal lavage fluids taken at different time points from the mice treated with terbinafine were significantly higher than those taken at corresponding time points from mice treated with fluconazole or itraconazole (P<0.01). The fungal burdens in the vaginal lavage fluids taken from mice 1 week after the beginning of the treatment with terbinafine remained at a relatively high level. A dramatic drop in the fungal burden was noted in the vaginal lavage fluids taken on the 2nd day of the treatment from mice treated with itraconazole or fluconazole group and the fungal burden on the 3rd day of the treatment in these mice were at a very low level, suggesting that fluconazole or itraconazole were highly effective for the treatment. However, the difference in the therapeutic effect between the two drugs was not significant (P>0.05). Itraconazole or fluconazole, but not terbinafine, is very effective for the treatment of fungal vaginitis caused by C. albicans in mice.

Establishment of lymphangioma model and a study on the promoting effect of murine melanoma cell B16-F1 on the lymphangiogenesis in vitro.

SummaryTo establish an animal model of benign lymphangiomas of C57BL/6 mouse in vitro and to observe the effect of mouse ascites melanoma cell B16-F1 on the lymphangiogenesis, 16 C57BL/6 mice aged 8 weeks were given two intraperitoneal injections of incomplete Freund’s adjuvant at a 15-day interval. The induced neoplasms were studied histopathologically and thhe neoplasms specimens were immunohistochemically examined for the expressions of VEGF-C (vascular endothelial growth factor-C) and Flt-4 (VEGFR-3, vascular endothelial growth factor receptor-3). The neoplasms were harvested and embedded in fibrin gel for culture in conditioned medium of B16-F1 cells in vitro and observed under inverted microscope. Our results showed that white solid tumor masses developed in peritoneal cavity after the induction. The tumors were confirmed to be lymphangioma by gross and histological examination. The tumor cells expressed both VEGF-C and Flt-4. Lymphatic capillaries coming from lymphangioma specimen grew into the gel and the conditioned medium of B16-F1 cells was found to be able to promote the growth of the vessels. It is concluded that intraperitoneal injection of incomplete Freund’s adjuvant is a good method for inducing benign lymphangiomas in mouse and B16-F1 cells can promote lymphangiogenesis.To establish an animal model of benign lymphangiomas of C57BL/6 mouse in vitro and to observe the effect of mouse ascites melanoma cell B16-F1 on the lymphangiogenesis, 16 C57BL/6 mice aged 8 weeks were given two intraperitoneal injections of incomplete Freund’s adjuvant at a 15-day interval. The induced neoplasms were studied histopathologically and thhe neoplasms specimens were immunohistochemically examined for the expressions of VEGF-C (vascular endothelial growth factor-C) and Flt-4 (VEGFR-3, vascular endothelial growth factor receptor-3). The neoplasms were harvested and embedded in fibrin gel for culture in conditioned medium of B16-F1 cells in vitro and observed under inverted microscope. Our results showed that white solid tumor masses developed in peritoneal cavity after the induction. The tumors were confirmed to be lymphangioma by gross and histological examination. The tumor cells expressed both VEGF-C and Flt-4. Lymphatic capillaries coming from lymphangioma specimen grew into the gel and the conditioned medium of B16-F1 cells was found to be able to promote the growth of the vessels. It is concluded that intraperitoneal injection of incomplete Freund’s adjuvant is a good method for inducing benign lymphangiomas in mouse and B16-F1 cells can promote lymphangiogenesis.

Construction of prokaryotic expression plasmid of mtrC protein of Neisseria gonorrhoeae and its expression in E. coli.

SummaryIn order to provide a rational research basis for detection of resistance ofNeisseria gonorrhoeae transferable resistance (mtr) efflux system, plasmid pET-28a(+) encoding mtrC gene was constructed and the related target protein was expressed inEscherichia coli (E. coli) DE3. The fragments of mtrC gene ofNeisseria gonorrhoeae from the standard strains were amplified and cloned into prokaryotic expression plasmid pET-28a(+) with restriction endonuclease to construct recombinant pET-mtrC which was verified by restriction endonuclease and DNA sequencing. The recombinant was transformed intoE. coli DE3 to express the protein mtrC induced by IPTG. The results showed mtrC DNA fragment was proved correct through restriction endonuclease and DNA sequencing. Its sequence was 99.5% homologus to that published on GeneBank (U14993). A 48. 5 kD fusion protein which was induced by IPTG was detected by SDS-PAGE. It was concluded that the construction of prokaryotic expression plasmid of mtrC protein ofNeisseria gonorrhoeae was correct and the fusion protein was successively expressed inE. coli.

In vitro Recombination and Identification of Mutated Fragment Corresponding to Regulation Region of mtrR Gene of Neisseria Gonorrhoeae

A site-directed mutant DNA fragment was synthesized and transfected into clinical Neisseria Gonorrhoeae (NG) stains to construct the transformants that contained the corresponding mutagenesis of regulation region of mtrR gene. According to the technique of gene splicing by overlap extension (SOEing), a DNA segment with specific mutagenesis was constructed by two-step polymerase chain reaction (PCR). The mutation fragments EF could be used for the next experiment in which the mutation NG strains were induced. By comparing the recombinant EF fragments to the corresponding DNA fragments of clinical NG strains, 2 of these were not compatible completely. The results of sequencing revealed that there was a 9 bp deletion between the 45 to 54 inverted repeat sequence localized within the mtrR promoter. It can be confirmed that the fragments EF are the specifically designed mutant fragments.

Relationship between mutation of IR in the mtr system of Neisseria gonorrhoeae and multiple antibiotic resistance.

SummaryTo study the relationship between mutation of the inverted repeat sequence (IR) in the multiple transferable resistant system (mtr) of Neisseria gonorrhoeae (NG) and its multiple antibiotic resistance, minimal inhibitory concentrations (MICs) for the clinically isolated strains were tested by agar-dilution-method. The mtr systems IR gene of NG was sequenced after amplification by polymerase chain reaction (PCR). Either two susceptive or five penicillin-resistant strains had no base mutation in IR gene, while all of the 13 strains with multiple-antibiotic-resistance had a single-base deletion (A/T). The result suggests that a single-base deletion of the thirteen-base IR sequence in mtr system of NG might result in multiple antibiotic resistance but is not associated with single antibiotic resistance.

Expression of MDM2 protein and mRNA in condyloma acuminata

SummaryIn order to investigate the role of murine double minute 2 (MDM2) in cell proliferation and carcinogenesis in condyloma acuminatum (CA), immunohistochemistry andin situ hybridization were used to detect the expression of MDM2 protein and mRNA in normal skin and skin lesions of CA of vulva. PCR was also used to detect HPV types. The results showed that in 32 observed CA specimens, the expression of MDM2 protein and mRNA was detected in 18(56.25%) and 22 (68.75%) respectively, while the co-expression of MDM2 protein and mRNA was found in 14. PCR results revealed that HPV6/11 and HPV16/18 subtypes were shown in 28(87.5%) and 4 (12.5%) respectively out of 32 CA specimens. Out of the 18 positive specimens expressing MDM2 protein. HPV6/11 subtypes were shown in 15 and HPV16/18 subtypes in 3. In 22 positive specimens expressing MDM2 mRNA. HPV6/11 subtypes were shown in 18 and HPV16/18 subtypes in 4. No expression of MDM2 protein and MDM2 mRNA was observed in normal skin. Our study indicated that the overexpression of MDM2 might be involved in malignant proliferation and carcinogenesis of CA

Correlation between the Expression of Fas, Bcl-2 in Peripheral Blood Lymphocytes and the LeveL of IFN-γ, IL-4 in Serum of Patients with Condyloma Acuminata

SummaryIn order to investigate the correlation between the expression of the apoptotic regulatory proteins (Fas, Blc-2) in peripheral blood lymphocytes (PBLC) and the level of IFN-γ and IL-4 in serum of the patients with condyloma acuminata (CA) in the immune pathogenesis of CA, indirect immunofluorescence labeling method of flow cytometer and solid sandwich ELISA method were performed for detecting the expression of Fas, Bcl-2 in PBLC and the level of IFN-γ and IL-4 in serum of 60 cases of CA. The results showed the expression level of Fas in PBLC of CA was significantly higher than in the normal control group, but the expression level of Bcl-2 was significantly lower (bothP<0.01). The level of IFN-γ in serum of CA was significantly lower than in the normal control group (P<0.01), but IL-4 was significantly lower (bothP<0.01). The expression of Fas in PBLC had a negative correlation with the level of IFN-γ in serum of patients with CA, but had a positive correlation with the level of IL-4: The expression of Bcl-2 had a positive correlation with the level of IFN-γ, but had a negative correlation with the level of IL-4. All the correlation coefficients had significant differerce byt test (P<0.01). It was suggested abnormal apoptosis in PBLC, the suppressed secretion of the TH1-associated cytokines (eg: IFN-γ) and the increased secretion of the TH2-associated cytokines (eg: IL-4) existed in the patients with CA and might play an important role in the immune pathogenesis of CA.