The ATP2C1 gene mutation in one case of familial benign chronic pemphigus was investigated. One patient was diagnosed as familial benign chronic pemphigus by pathology, ultrastructral examination and clinical features. Genomic DNA was extracted from blood samples. Mutation of ATP2C1 gene was detected by polymerase chain reaction (PCR) and DNA sequencing. The results showed that deletion mutation was detected in ATP2C1 gene in this patient, which was 2374delTTTG. No mutation was found in the family members and normal individuals. It was concluded that the 2374delTTTG mutation in ATP2C1 gene was the specific mutation for the clinical phenotype for this patient and was a de novo mutation.
Calcium-Transporting ATPases/*genetics ; DNA Mutational Analysis ; Pemphigus, Benign Familial/*genetics ; Sequence Deletion

Chronic myelogenous leukemia (CML) has a high proportion in hematologic malignancy.Past decade,the appearance and development of tyrosine kinase inhibitors (TKIs) is the milestone of the treatment of CML.TKIs have antitumor effect with inhibition of the phosphorylation of kinases and the downstream signal transduction.In recent years,many large medical institutions at home and abroad worked on clinical experiment researches to investigate the mechanism of TKIs and how to choose TKIs in CML patients.This work is of great significance to the clinic.

To investigate the role of progesterone receptor (PR) expression in malignant melanoma (MM), PR and proliferative cell nuclear antigen (PCNA) expression were immunohistochemistrically evaluated in a series of 35 specimens of MM, and the correlation between the immunohistochemistrical findings and clinicopathological data was also analyzed. PR expression was detected in 25.7% (9/35) of the patients with MM. No PR expression was observed in nevi. PR expression was inversely correlated with PCNA expression (r=-0.353, P=0.026). PR expression was slightly increased in females, subjects aged under 55 y, those with ulceration, non-acral subtype and diagnosis delay longer than 1 y, but the difference was not statistically significant. Selective expression of progesterone receptor in malignant melanoma might be correlated with inhibited tumor growth.
Gene Expression Regulation, Neoplastic ; Immunohistochemistry ; Melanoma/*metabolism ; Models, Biological ; Prognosis ; Proliferating Cell Nuclear Antigen/*metabolism ; Receptors, Progesterone/*biosynthesis ; Receptors, Progesterone/genetics ; Skin/metabolism ; Skin Neoplasms/metabolism

Objective To study the effects of short hairpin RNA (shRNA) targeting STAT3 gene on the proliferation and apoptosis of A375 human malignant melanoma cells.Methods Sense and antisense oligonucleotides with small hairpin structures targeting STAT3 gene were designed,synthesized and cloned into the plasmid vector psiRNA-hHlneo after annealing.Cultured A375 cells were divided into 3 groups: control group receiving no treatment,psiRNA-H1 group transfected with empty plasmid,and psiRNA-H1/STAT3 group transfected with the recombinant plasmid containing the shRNA.After additional culture for different durations,reverse transcription PCR and Western blot were performed to detect the expression of STAT3 mRNA and protein,methyl thiazolyl tetrazolium (MTT) assay to evaluate cell proliferation,flow cytometry to assess cell cycle and apoptosis.Results The expression level of STAT3 mRNA and protein in A375 cells in psiRNA-H1/STAT3 group (0.2136 ± 0.0626,0.8214 ± 0.043,respectively) were significantly lower than that in the control group (0.7826 ± 0.0701,3.1693 ± 0.0846,respectively,both P ＜ 0.01) and psiRNA-H1 group (0.8518 ± 0.0783,3.218 ± 0.078,respectively,both P ＜ 0.01 ).The inhibition rates of cell proliferation at 24,48 and 72 hours were 21.35％ ± 2.12％,32.52％ ± 2.64％ and 40.4％ ± 3.08％ respectively in psiRNA-H1/STAT3 group,statistically different from those in the control group (1.39％ ± 0.53％,3.05％ ± 1.16％,4.41％± 1.42％,respectively,all P ＜ 0.01) and psiRNA-H1 group (2.63％ ± 1.38％,5.84％ ± 2.47％,10.32％ ±2.48％,respectively,all P ＜ 0.01).Flow cytometry showed a statistical increase in cell apoptosis rate in psiRNA-H1/STAT3 group compared with the control and psiRNA-H1 group (81.06％ ± 2.10％ vs.26.28％ ±0.47％ and 27.31％ ± 1.05％,both P ＜ 0.01 ).The psiRNA-H1/STAT3 group exhibited a higher percentage of cells at G0/G1 phase (68.43％ ± 4.00％) but a lower percentage of cells at S phase (17.4％ ± 2.05％) compared with the control group (60.07％ ± 2.47％,P ＜ 0.05; 28.40％ ± 2.09％,P ＜ 0.01 ) and psiRNA-H1 group (60.29％ ± 2.26％,27.34％ ± 3.63％,both P ＜ 0.05 ).Conclusions The small interference RNA targeting STAT3 gene can specifically down-regulate the expressions of STAT3 mRNA and proteins in,inhibit cellular proliferation of,and induce apoptosis in,A375 cells.

Objective To summarize the management of cardiopulmonary bypass(CPB) for infants below 10 kilograms with congenital heart diseases.Methods From Jan 2010 to Apr 2011,the clinical datas of 122 infants aged from 1.5 months to 2 years with body weight 3 to 10 kilograms,who underwent open heart surgery under mild or moderate hypothermia CPB were retrospectively analyzed.Results Among all the 122 infants,CPB time was 13 ～ 118 min [(62.69 ± 21.48) min],aortic cross-clamp time was 0 to 86 min [(35.47 ± 19.51) min].All patients were spontaneous resuscitation and successfully weared from the machine,no severe complications associated with CPB occurred,3 infants died (2.46％,3/122) after operation.Conclusion Using membrane oxygenator,circuit tubing and artery filter with less priming,and highflow perfusion during CPB,maintaining hemodynamics stable,holding reasonable hematocrit and colloid osmotic pressure,good myocardial protection and ultrafiltration are the significant elements of the management of CPB in infants with weight less than 10 kilograms.

In order to investigate the role of Th17 cytokines in the pathogenesis of psoriasis, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of IL-17, IL-23 (p19/p40), and IL-6 in skin lesions and non-lesions of the patients with psoriasis and skin tissues of normal subjects. The results showed that the mRNA expression levels of IL-17, IL-23p19, IL-23p40 and IL-6 in psoriasis lesion were significantly higher than those of non-lesions (1.231 +/- 0.843 vs 1.003 +/- 0.044, 1.166 +/- 0.142 vs 0.765 +/- 0.133, 1.125 +/- 0.104 vs 0.730 +/- 0.103, 1.186 +/- 0.222 vs 0.976 +/- 0.122, respectively, all P < 0.05). Meanwhile, The expression levels of IL-17 mRNA, IL-23p19 mRNA, IL-23p40 mRNA and IL-6 mRNA were higher in non-lesions than those in normal skin tissues (1.003 +/- 0.044 vs 0.620 +/- 0.104, 0.765 +/- 0.133 vs 0.584 +/- 0.078, 0.730 +/- 0.103 vs 0.000 +/- 0.000, 0.976 +/- 0.122 vs 0.656 +/- 0.121, respectively, all P < 0.05). The overexpression of Th17 cytokines in the skin lesions of patients with psoriasis may indicate that Th17 cytokines play a very important role in the immunopathogenesis of psoriasis.

To investigate the role of toll-like receptor 9 (TLR9) in the pathogenesis of lichen planus, the expressions of TLR9 and its mRNA in the lesional skin of lichen planus were detected by immunohistochemical technique (SP) and RT-PCR. As control, normal skin of healthy volunteers was also tested. The immunohistochemical study showed that the expression of TLR9 in the lesional skin of lichen planus was significantly higher than that in the normal controls. The results of RT-PCR showed that both skin lesions and normal controls had TLR9 expression. In skin lesions, the expression level of TLR9 mRNA was 1.6075+/-0.0930, which was significantly higher than that in normal controls (P<0.001). These findings indicated that up-regulated expression of TLR9 and its mRNA might be involved in the pathogenesis of lichen planus.

To investigate the expression of vaginal Th1 and Th2 cytokines in rats with experimental vaginal candidiasis under different immune conditions, ICR murine vaginal candidiasis model was established and immno-suppressed murine models of vaginal cadidiasis were established in estrogen-treated mice. Non-estrogen-treated mice were used as controls. The mRNA level of Th1 (IL-2)/Th2 (IL-4, IL-10, TGF-beta1) cytokines in murine vaginal tissues was determined by RT-PCR. The cykotine in local tissues was increased to different extent under normal immune condition. IL-2 mRNA was increased during early stage of infection, while IL-10 was increased transiently during late stage of infection. TGF-beta1 production was found to be increased persistently. At same time, the expression of IL-2 mRNA was suppressed in immno-suppressed group, and the level of IL-4, IL-10, and TGF-beta1 were higher than the normal immunity group to different degree during infection. The high level of IL-2 mRNA during early stage of infection was associated with clearance of mucosal Candidia albicans (C. albicans), and its expression suppressed leading to decreased clearance of mucosal C. albican in immuno-suppression. The over-expression of IL-4 and IL-10 could significantly enhance the susceptibility to C. albicans infection in mice.

In order to investigate the role of MMP-9 and TIMP-1 in the pathogenesis of systemic sclerosis, the expression of MMP-9 and TIMP-1 was immunohistochemically detected in skin lesions of the patients with diffuse cutaneous systemic sclerosis, skin lesions of the patients with limited cutaneous systemic sclerosis, and skin tissues of normal subjects. The results showed that the expression of MMP-9 in lesions of diffuse cutaneous systemic sclerosis was significantly lower than that of normal skins (P<0.05). However, no significant difference in the level of MMP-9 in the limited cutaneous systemic sclerosis and normal skin was found. Meanwhile, the expression of TIMP-1 in lesions of diffuse cutaneous systemic sclerosis and limited cutaneous systemic sclerosis were significantly higher than that of normal skins (both P<0.05). It was suggested that the expression of MMP-9 and TIMP-1 might play an important role in the development of systemic sclerosis.

In order to investigate the association of genotypes of HLA-DRB1 and HLA-DQB1 alleles with the genetic susceptibility of chronic urticaria (CU), genotypes of HLA-DRB1 and HLA-DQB1 genes were detected by polymerase chain reactions with sequence-specific primers (PCR-SSP) in 42 patients with CU (19 men and 23 women, mean age 30.67+/-12.45 y old as well as 193 racially matched healthy persons in ethnic Han from Hubei provinece. Gene frequencies of HLA-DRB1 * 12, * 0901 (RR=3.11, chi2=7. 579, P=0.006; RR= 2.47, chi2 =5.684, P=0.017) were significantly increased in CU patients as compared with that in healthy people. Gene frequencies of HLA-DQB1 * 05 (RR=0.26, chi2=6.683, P=0.01) were significantly decreased in CU patients. It was suggested that CU was found strongly associated with HLA-DRB1 * 12, * 0901 and HLA-DQB1 * 05, the former might be the genetic markers for susceptibility to CU, but the latter might play a resistive role.
Alleles ; Chronic Disease ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; HLA-DQ Antigens/*genetics ; HLA-DR Antigens/*genetics ; Polymerase Chain Reaction ; Urticaria/*genetics