Yi M

Depletion of CD147 sensitizes human malignant melanoma cells to hydrogen peroxide-induced oxidative stress.

BACKGROUND Increased sensitivity to reactive oxygen species (ROS) contributes to the effectiveness of therapeutic strategies in patients with malignant melanoma (MM). CD147, a cell surface receptor for cyclophilin A (CypA), is thought to exert antioxidant activities. OBJECTIVE To understand the influences and mechanisms of CD147 on proliferation, apoptosis and redox state of A375 cells under H(2)O(2)-induced oxidative stress. METHODS The effect of CD147 silencing on cell viability, apoptosis, the generation of ROS, superoxide dismutase (SOD) activity, and the malondialdehyde (MDA) level that reflects oxidative damage, was measured in human malignant melanoma cell line A375 treated or untreated with hydrogen peroxide (H(2)O(2)). RESULTS In A375 cells, CD147 silencing increased the H(2)O(2)-induced inhibition of cell viability, H(2)O(2)-induced apoptosis, H(2)O(2)-mediated ROS- and MDA generation, and the H(2)O(2)-triggered decrease in SOD activity. CONCLUSIONS Our results demonstrated that CD147 silencing increased cellular ROS and destroyed the intrinsic antioxidant defenses in A375, indicating that CD147 exerts a cytoprotective effect against H(2)O(2)-induced oxidative damage.

Activation of JAK-STAT1 signal transduction pathway in lesional skin and monocytes from patients with systemic lupus erythematosus.

OBJECTIVE To study the activation of Janus protein tyrosine kinase (JAK)/signal transducer and activator of transcription 1 (STAT1) signaling pathway and its inhibitor-signal transducer and activator of transcription-1(SOCS-1) in patients with systemic lupus erythematosus. METHODS A total of 45 patients with active systemic lupus erythematosus (SLE) and 30 healthy controls were randomly selected. Western blot was performed to measure the expression of Stat1 protein and phospho-Stat1 protein (an activated form of Stat1 protein) in the monocytes after stimulation with recombinant high mobility group box1 (rHMGB1) at various time points. Expression of Stat1 protein in the skin or lesional skin was also detected. Phasic expressions of SOCS-1 mRNA in the monocytes after rHMGB1 stimulation were detected by real-time reverse transcription-polymerase chain reaction. SOCS-1 gene expression in the skin or lesional skin was also detected. RESULTS The expression level of Stat1 proteins in the monocytes from patients with SLE was higher than that from healthy controls (t=9.16, P<0.01) and positively correlated with SLE disease activity index (SLEDAI) (r=0.59, P<0.01). Expression of phospho-Stat1 in the monocytes from SLE patients was time-dependently upregulated after stimulation with rHMGB1 at various time points, while expression of SOCS-1 mRNA remained unchanged (all P>0.05). Expressions of phospho-Stat1 protein and SOCS-1 mRNA in the monocytes from healthy controls were increased transiently after stimulation with rHMGB1 (all P<0.05). Both expressions of phospho-Stat1 protein and SOCS-1 gene in the lesional skin from patients with SLE were upregulated compared with those in normal skin from healthy controls (all P<0.01). CONCLUSION There are hyperactivation of JAK-STAT1 signaling pathway and negative feedback down-regulation of SOCS-1 in patients with systemic lupus erythematosus. HMGB-1 may be partly involved in the pathogenesis of SLE by the abnormal mediating function of JAK-STAT1 signal transduction pathway.

Distribution and expression of alternative splice isoforms of NOR1 in human tissues and cell lines.

OBJECTIVE To analyze the distribution and subcellular localization of NOR1 alternative splice isoforms in human tissues and cell lines. METHODS NOR1 open reading frame(ORF) was amplified from human fetal brain cDNA library and subcloned into pCMV/myc vector. The level of NOR1 mRNA in human tissues was determined by real-time RT-PCR. Region spanning exon 2 was amplified from cDNA or genomic DNA by specific primers and sequenced. The expression plasmids of NOR1 were transfected into cells and immunofluoresence assay was performed to determine the subcellular localization of NOR1 protein isoforms in human cells. RESULTS The expression of NOR1 mRNA was high in human adult testis, moderate in human fetus nasopharynx, trachea, brain, and kidney tissues, and weak or undetectable in other tissues. Two splice variants of human NOR1 gene resulted from alternative splicing at exon 2 were identified. Both of isoform 1 and isoform 2 mRNA were detected in human fetus brain. Isoform 2 was the sole isoform in other tissues but brain. Only isoform 2 mRNA was detected in cell lines used in this study, though no genomic deletion of exon 2 could be found in all these cell lines. Immunofluoresence assay showed both isoform 1 and isoform 2 proteins were distributed in cytoplasm. CONCLUSION Alternative splice isoforms of tumor suppressor gene NOR1 are identified. NOR1 isoform 1 and isoform 2 are both detected in fetus brain. NOR1 isoform 2 lacking of exon 2 is the sole isoform in multiple tissues except for brain. The exon 2 encoded peptide does not affect the subcellular location of NOR1 protein.

Expression of cyclophilin A and CD147 during skin aging.

OBJECTIVE To investigate the role of cyclophilin A (CypA) and CD147 in the process of skin aging. METHODS Twenty cases of tissue samples from junior group(<15 years old), middle age group(30-40 years old)or old age group (>65 years old) were collected from photophobic and exposal parts of skin, respectively. Immunohistochemistry (IHC) and in situ hybridization (ISH) were carried out to semi-quantitatively detect the expression level of CyPA and CD147. RESULTS IHC demonstrated that both CyPA and CD147 were expressed in both photophobic and exposal parts of normal human skin in all 3 groups. The expression levels of both CyPA and CD147 were increased with increase in age. There were significant differences in both CyPA and CD147 expression among 3 groups (P<0.05). CyPA and CD147 were also positively correlated in all 3 groups. Similar results were achieved by ISH. CONCLUSION The interaction between CD147 and CyPA might play an important role in the process of skin aging.

A survey on AIDS knowledge rate and sexual behavior among men who have sex with men population at sexually transmitted disease clinic

OBJECTIVE To survey on men who have sex with men (MSM) populations sexual behaviors, condom-service condition, HIV related knowledge and other issues among MSM population at sexually transmitted disease (STD) clinics to understand the characteristics of behaviors and offer suggestions for effective health education and behavior intervention. METHODS From January to December, 2009, we used anonymous questionnaires which involved in their mastery of demographic characteristics, sexual behavior, condom-service condition, HIV related knowledge, and so one, to investigate 200 MSM at 3 STD clinics of comprehensive hospital. RESULTS The average age of informant was (26.7+/- 8.9) years and 121 individuals (62.6%) had confirmed with STD in recent one year. In the recent 6 months, the average number of homosexual partners was 9.2+/- 4.8 and 102 (52.8%) had heterosexual partners. In the sexual intercourse with homosexual, 123 individuals had anal intercourse (63.7 %) and 117 had oral intercourse (60.6%). In the sexual intercourse with heterosexual, 92 (90.2%) individuals had vaginal intercourse, 37 (36.2%) had anal intercourse, and 59 (57.8%) had oral sex behavior.There were a statistical difference between heterosexual and homosexual sex behaviors (P<0.01). The condom-using frequency had statistic difference in different sexual behaviors(χ²=188.396, P<0.001). There was no linear correlation between HIV related knowledge and condom-using condition in sexual behaviors except the heterosexual anal intercourse. CONCLUSION High AIDS knowledge mastery rate is found in our survey. The respondents get HIV/AIDS knowledge through various ways actively. There is no obvious relation between the mastery of HIV related knowledge and condom-using frequency. The ratio of non-protected sexual behaviors is high in heterosexuals. How to adopt effective methods for behavior intervention to MSM at STD clinic needs to be further studied.

[Metabolic reprogramming in cancer: the art of balance].

Despite seminal studies in the 1920s by Warburg showing a characteristic metabolic pattern for tumors, cancer bioenergetics has often been relegated to the backwaters of cancer biology. Recent studies have shown that metabolism in the tumor tissue is far more complicated than we previously knew. Despite vigorous glycolysis, in fact, the tumor tissue still retains mitochondrial aerobic metabolism. Mitochondria is one of the main sites of the biosynthesis process of tumor cells. Recent studies revealed that abnormal fatty acid metabolism. Amino acid metabolism plays a key role in the tumorigenesis. The metabolic network in tumor cells was reprogrammed, leading to nutrition flux reorganization and re-direction. Metabolism reprogramming in tumor cells facilitates the balance between the needs of energy supply and the synthesis of biological macromolecules. Targeting cell metabolism is not intended to interfere the energy supply of tumor cells but to affect the synthesis of metabolic rate, thereby inhibiting the proliferation of the tumor. The review focuses on the importance of metabolic reprogramming in tumor development and cancer therapy. We summarize what is currently known about metabolic reprogramming and establish a framework to understanding its contribution to the altered metabolism of cancer cells.

Characteristics of health seeking behaviors for patients with psoriasis in a general hospital

OBJECTIVE To explore the characteristics of health seeking behaviors and relevant factors among the outpatients with psoriasis in a general hospital, and to provide scientific evidence to modify the inappropriate health seeking behaviors for psoriasis patients and improve the therapeutic effect. 
 METHODS Using cross-sectional study design, a dermatological clinic in a general hospital was selected for this study. Two hundred psoriasis outpatients from 1st April to 30th September in 2014 were enrolled. A series of questionnaires were used to collect information regarding the social-demographic characteristics, health seeking behaviors, knowledge on psoriasis, and depressive and anxious symptoms.
 RESULTS Among 200 psoriasis outpatients, 141 patients (70.5%) experienced transfer treatment. One hundred and eighty-two patients (91.0%) went to formal hospitals in county for treatment when symptoms were firstly displayed; the other 18 patients (9.0%) went to village or community clinics, or the private clinics. Over 50% patients chose municipal hospital and 35.0% patients went to provincial hospital for medical treatment among the 182 patients. Ninety-one percent of the patients made decision for the first treatment after consulting with their spouses or relatives. According to the definition of the appropriate health seeking behavior in this study, 50 patients (25%) had inappropriate health seeking behaviors. Logistic regression analysis showed inappropriate health seeking behaviors correlated with the family whose income was less than 3 000 yuan per person per month (OR=2.232, 95%CI 1.086 to 4.585), patients who made the decision for the treatment with no discussion with other people when the symptoms appeared (OR=3.016, 95%CI 1.023 to 8.893), and the high score of the International Psoriasis Severity Index (OR=1.043, 95%CI 1.001 to 1.088). 
 CONCLUSION The dermatological practitioner should pay more attention to the factors, such as social-demographical characteristics, especially the low level of family income and their treatment experience. It is necessary for the general hospital to conduct health education for psoriasis outpatients to promote their appropriate health seeking behaviors.

Effect of NOR1 gene knockdown on the biological behavior of HeLa cells

OBJECTIVE To explore the effect of the oxidored nitro domain containing protein 1 (NOR1) gene knockdown on the biological behavior of HeLa cells in cervical carcinoma. METHODS The recombinant plasmids pSUPER-shNOR1-1, pSUPER-shNOR1-2 and pSUPERscramble, which targeted to NOR1 gene, were constructed by pSUPER.neo+GFP vector, transfected into HeLa cells respectively using Lipofectamine 2000 reagent, and followed by G418 selection. The expression level of NOR1 mRNA and protein were determined by RT-PCR and Western blotting, respectively. Methyl thiazolyl tetrazolium (MTT) assay was performed to determine the growth curve of cell viability. The stable transfectants were treated with H₂O₂ and cell apoptosis was determined by Hoechst 33258 staining and terminal deoxynucleotidyl transferasemediated dUTP nick end labeling (TUNEL) assay. The expression levels of Bcl-2, cleaved caspase 9 and poly ADP-ribose polymerase (PARP) were measured by Western blot. RESULTS NOR1- knockdown HeLa cells were successfully constructed by transfection of pSUPER-shNOR1-1 or pSUPER-shNOR1-2 plasmids into HeLa cells. MTT assay showed that the silence of endogenous NOR1 in HeLa cells could lead to the increase in cell viability and proliferation, and the inhibition of H₂O₂-induced apoptosis compared with the negative control. Western blot showed that the expression level of active caspase 9 and cleaved PARP was inhibited in NOR1-knockdown cells when they were treated with H₂O₂ while the expression level of Bcl-2 protein increased. CONCLUSION Silence of endogenous NOR1 facilitates the cell viability and growth of HeLa cells, and attenuates HeLa cells apoptosis induced by H₂O₂, which might be mediated by up-regulation of Bcl-2 level and down-regulation of the cleaved caspase 9 cascade.

Effect of DNA hypermethylation on NOR1 promoter activity and expression

OBJECTIVE To analyze the effect of DNA hypermethylation on NOR1 promoter activity and expression. METHODS NOR1 promoter plasmids were treated with SssI methyltransferase. The plasmids were modified by sodium bisulfite and purified. Sodium bisulfite-modified plasmids were subjected to PCR with primers designed to analyze the methylation status of 26 CpG sites in a 311-bp region of the NOR1 promoter. Cells were transfected by methylated or mock-methylated promoter plasmids. The promoter activities were assessed by the luciferase levels of cell lysates or by directly observing GFP expression under fluorescence microscope. HL60 cells were treated with different concentrations of 5-aza-dC. Total RNA was isolated from harvested cells. Real-time RT-PCR was used to measure the expression level of NOR1 mRNA. RESULTS Bisulfite sequencing confirmed that SssI methyltransferase treatment successfully resulted in intensive hypermethylation of the NOR1 promoter plasmids. The promoter activity of NOR1 promoter plasmids was totally blocked by SssI methyltransferase treatment. NOR1 expression levels in HL60 cells were restored by 5-aza-dC treatment. CONCLUSION NOR1 promoter plasmids are intensively hypermethylated by SssI methyltransferase treatment. The promoter activity of NOR1 promoter plasmids are totally blocked by SssI methyltransferase treatment. The 5-aza-dC treatment may restore the endogenous NOR1 mRNA level in HL60 cells.

Lichen planus in Hunan:124 patients

OBJECTIVE To determine the clinical feature of lichen planus in Hunan, and to provide references for correct diagnosis and treatment. METHODS The clinical data from 124 outpatients with lichen planus attending Dermatology Department of Xiangya Hospital, Central South University from January 2000 to April 2008 were analysed retrospectively, including data on sex, age of visit or onset, course of disease, clinical manifestation, histopathology, and so on. RESULTS In these 124 patients, male to female ratio was 1:1. Most patients were in 30-60 age group when they first visited the Out-patient Department. There were 2 onset peaks, one in 14-29 age group and the other in 40-60 age group. Most (35.5%) of the lesions were chronic localized subtypes. The limbs were the most common site to be involved, and they were also the most prevalent site for onset of lichen planus(74 cases, 59.7% ). The second common site was the trunk (29 cases, 23.4%). Fifteen patients (12.1%) had genitalia lesions, and 9 (7.3%) accompanied with oral mucosal involvement. Most of the eruptions were half-distributed (44.3%), 64 patients (51.6%) had pruritus,which ranged from mild irritation to severe intolerable itching, 56 (45.2%) had no subjective symptoms, 4 (3.2%) had spontaneous pain or tenderness, and 2 had positive family history of lichen planus. The diagnostic accordance rate was 50.8%. CONCLUSION Some characteristics exist in sex, age, and clinical manifestation of lichen planus in Hunan province. Systemic and/or local treatment should be taken according to the lesion morphology, localization and subtypes of lichen planus.