Cross-sectional study on clinic behavior and therapeutic status of patients with psoriatic arthritis in multi-center / 北京大学学报(医学版)

OBJECTIVE@#To investigate and analyse the features of treatment behavior and standardized therapeutic status of patients with psoriatic arthritis (PsA).@*METHODS@#Out patients diagnosed with PsA in People's Hospital of Peking University, Haidian Hospital, People's Hospital of Jianyang City, Central Hospital of Xinxiang City, Integrated Traditional Chinese and Western Medicine Hospital of Cangzhou City, The Third Hospital of Hebei Medical University from February to June 2018 were enrolled in this investigation. The data including gender, age of onset, course of disease, site of first consulting department, time of the first visit and definite diagnosis, follow-up interval, and use of conventional disease modifying anti-rheumatic drugs (cDMARDs) and biological DMARDs (BioDMARDs) were collected and analyzed.@*RESULTS@#In the cross-sectional study, 133 PsA patients were investigated. The mean age of onset was (47±11) years, the male to female ratio was 1.3:1, and mean disease duration was (16±8) years. Rheumatology department was the most common site of first hospital visit (37.6%, 50/133). Orthopedics department and dermatological department were visited by 24.1% (32/133) and 23.3% (31/133), respectively. Ratio of definite diagnosis was the highest in rheumatology department which was 78% (39/50). The ratio of definite diagnosis of dermatological department was the second highest, which was 19.4% (6/31). The mean definite diagnosed time was 7.6 months since the first visit of PsA patients, and diagnosed time was the shortest in rheumatology department, which had statistical significance. 37% PsA patients were treated appropriately in 3 months, 17.3% PsA patients were treated in 3-6 months and 40.2% patients with PsA visited their doctor more than once a year. 48.8% patients hadn't received standardized treatment before visit, and one third patients never received the therapy of DMARDs. Methotrexate was the most commonly used cDMARDs (58.3%), followed by leflunomide (20.5%) and BioDMARDs (19.7%), and biologicals were tumor necrosis factor antagonists.@*CONCLUSION@#In this multi-center study, the first visit department of PsA patients was widely distributed, and most patients were definitely diagnosed in Rheumatology Department. The time of their first visit and definite diagnosis were delayed due to multi factors. Nearly half of the patients did not receive standardized treatment.

The level of chemokine C-C motif ligand 2 in systemic lupus erythematosus patients and its association with the 2518 A/G polymorphism in the CCL2 promoter region / 中华风湿病学杂志

Objective To explore the correlation between chemokine C-C motif ligand 2 (CCL2)-2518A/G polymorphism and the incidence of systemic lupus erythematosus (SLE),and investigate the effects of CCL2-2518A/G polymorphism on the expression of CCL2.Methods A total of 134 SLE patients and 56 sex and age matched healthy people were enrolled in this study.CCL2 plasma levels were measured by enzymelinked immunosorbent assay(ELISA).The 2518 A/G polymorphism in CCL2 promoter region was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP),and then peripheral blood mononuclear cells (PBMCs) were cultured in vitro to explore the influence of the 2518A/G polymorphism in CCL2 promoter region on the expression of CCL2.Mann-Whitney U-test,Student's t test and chi-squared test were used for analysis.Results The plasma CCL2 level in the SLE group was 358.5 (500.1) pg/ml [median (four interval)],which was significantly higher than that in the control group 243.6(125.8) pg/ml (Z=3.892,P=0.000).Patients with high plasma CCL2 levels were more prone to have renal (x2=7.159,P=0.007) and der-matomucosal (x2=5.133,P=0.023) involvement,as well as much higher disease activity index (SLEDAI) scores (Z=2.012,P=0.047).The results of the analysis of individual CCL2-2518A/G polymorphic genotypes suggested that patients with the G/G genotype had the highest CCL2 levels 425.7 (608.8) pg/ml,followed by those with the G/A genotype 355.3(511.1) pg/ml and A/A genotype 327.8(367.9) pg/ml (x2=3.496,P=0.048).The results of in vitro experiment showed that after the stimulation of lipopolysaccharide,the expression of CCL2 in PBMCs with G/G homozygote increased more significantly than that with A/A homozygote (P＜0.05).Conclusion The increase of plasma CCL2 concentrations is associated with tissue injury and high SLEDAI,which suggests that CCL2 may play a crucial role in the pathogenesis of SLE.CCL2-2518A/G polymorphism is not related to the pathogenesis of SLE,but it can affect the condition of SLE by promoting the expression of CCL2 in inflammatory environment.

Pregnancy investigation in idiopathic inflammatory myopathy in China / 中华风湿病学杂志

Objective We investigated female patients with idiopathic inflammatory myopathy (ⅡM),including polymyositis (PM) and dermatomyositis (DM) to understand if the activity and treatment of ⅡM affected the fetus outcomes as well as the correlation between pregnancy and the pathogenesis of ⅡM.Methods Questionnaires were designed,including patients' fertility,the activity and treatment of ⅡM during pregnancy and after childbirth,and the fetal outcomes.The questionnaires were conducted on 75 female patients (22-53 years old),who once hospitalized at Peking Union Medical College Hospital from 2007 to 2013.The mutual effect between the ⅡM disease activity,pregnancy and the fetus were compared by Chi square.Results Seventy-five patients (19 PM,56 DM) had 144 pregnancies,but only 18 of them became pregnant after the onset of the disease.There were two patients had onset of DM during pregnancy,and two patients suffered from DM after delivery or abortion within one month.The rates of abnormal pregnancy in patients with pregnancy before the onset of ⅡM was significantly lower than those with pregnancy after the onset of ⅡM (x2=10.21,P=0.001).Of the 18 pregnancies after the onset of ⅡM,7 had active ⅡM during pregnancy,3 of them flareddue to pregnancy.The 5 abnormal pregnancies (5/7) were seen in seven pregnancies totally,including two spontaneous abortion,one embryo diapause and two premature births.While of the other 11 pregnanciesin stable condition had only 2 abnormal pregnancies (2/11),including one fetus ended prematurely and one embryo diapause.The fetal outcomes were significantly related to the status of illness (x2=5.103,P=0.024).Two patients became pregant after the onset of the disease,both of them had stable disease before pregnancy.However,the disease became active during the pregnancies.One of them flared with rash during two pregnancies.The fetal end in embryo diapause and premature birth (33+4 weeks),respectively.Another flared with muscle weakness at five months of gestation.The fetal ended in premature birth (36+3 weeks).Two patients had onset of DM within one month following the delivery and voluntary abortion.Patients with active disease had taken medium to large doses of prednisone (30-60 mg) orally,in combination with hydroxychloroquine (0.2 bid) or without.In addition,two patients administrated with intravenous gamma globulin (IVIG) at low dose,whose illness condition had been well controlled.Conclusion Pregnancies after the onset of ⅡM tend to end with spontaneous abortion,embryo diapause,premature birth.The fetal outcomes are significantly related to disease activity.When ⅡM flared during pregnancy,IVIG might be the most effective and safe treatment,combined with corticosteroids.

Investigation on the mechanisms of p15INK4B gene demethylation by valproate in Molt-4 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the antitumour effects of sodium valproate (VPA) on the proliferation, differentiation and cell cycle of Molt-4 cell and to investigate its demethylation mechanisms.</p><p><b>METHODS</b>After Molt-4 cells trated with VPA at different concentrations, cell viability and growth curve were assessed by MTT assay. Cell cycle changes were analyzed by flow cytometry. The expression level of p15, DNA methyltransferase 1 (DNMT-1), DNMT3A and 3B mRNA were detected by RT-PCR and the methylation level was detected by hn-MSPCR.</p><p><b>RESULTS</b>VPA significantly inhibited the proliferation of Molt-4 cells. After 48 h culture with 5.0 mmol/L VPA, the percentages of Molt-4 cells in G(0)/G(1) phase was (66.87 ± 3.31)% and in S phase was (8.47 ± 2.56)%, while in control group, the cells in G(0)/G(1) phase increased and in S phase decreased significantly. The p15 gene in Molt-4 cells failed to express due to its hypermethylation. The expression level of p15 gene mRNA increased significantly after exposure to VPA for 48 h. As compared with control group, the expression of DNMT-1 was down-regulated in a dose-dependent manner. The expression level of DNMT3B decreased at 10.0 mmol/L concentration.</p><p><b>CONCLUSION</b>VPA has a demethylation effect on p15 INK4B gene by inhibiting the DNMT-1 and DNMT3B gene activities to recover p15 gene activity, which arrests Molt-4 cell in G(0)/G(1) phase.</p>

Inhibitory effect of VPA on multiple myeloma U266 cell proliferation and regulation of histone acetylation / 中国实验血液学杂志

This study was aimed to investigate the effects of sodium valproate (VPA) on the proliferation and regulation of histone acetylation of multiple myeloma cell line U266. U266 cells were treated with VPA. Cell proliferation was determined by CCK-8 assay, and cell cycle were analyzed by flow cytometry (FCM). The expression level of HDAC1 mRNA was detected by RT-PCR, and the protein levels of HDAC1 and histone H3, H4 acetylation was detected by Western blot. The results showed that the VPA inhibited the proliferation of U266 cells in concentration-and time-dependent manners.After exposure to different concentrations of VPA for 48 hours, the proportion of G(0)/G(1) cells increased, while the proportion of S phase cells decreased. The cell cycle was arrested obviously in G(0)/G(1) phase (p < 0.05). The expression of HDAC1 mRNA was inhibited, and the protein level of HDAC1 was down-regulated, while the histone H3/H4 acetylation was up-regulated in U266 cells. It is concluded that the VPA can inhibit cell proliferation of U266 and induce G(0)/G(1) phase arrest. The increase of histone H3/H4 acetylation resulting from inhibiting HDAC1 by VPA might be considered as a possible mechanism.

Demethylation and transcription of p16 gene in malignant lymphoma cell line CA46 induced by EGCG / 中国实验血液学杂志

The study was purposed to investigate the possible mechanism of epigallocatechin-3-gallate (EGCG) induced p16 gene demethylation and transcription regulation in the malignant lymphoma cell line-CA46. The induced growth inhibition of CA46 cells was assayed by growth curve and MTT; the DNA content of CA46 cells was analyzed by flow cytometry after being exposed to EGCG; the methylation status of the p16 gene in CA46 cell line before and after treatment with EGCG was detected by the nested-methylation specific PCR and DNA sequencing; the mRNA of p16 and DNA methyltransferases (DNMT3A and DNMT3B) gene were determined by RT-PCR. The results showed that in comparison with the control, all the 3 different concentration of EGCG were able to inhibit the growth of malignancy cell lines and increase the cell number in G(0)/G(1) phase. After treatment with EGCG for 48 hours, the methylation level was apparently attenuated in a concentration-dependent manner. Expression of p16 gene in untreated group was mild while in the treated groups it had been greatly strengthened, as compared with untreated group, the gray scale ratio of p16 to beta-actin 1 treated with EGCG (6, 12, 24) microg/ml was increased from (0.05 +/- 0. 01) to (0.19 +/- 0.03), (0.39 +/- 0.10), (0.85 +/- 0.09) respectively, exhibiting a significant difference (p < 0.05); as compared with the untreated group, after treatment with EGCG for 48 hours, the expressions of DNMT3A and DNMT3B were obviously down-regulated. It is concluded that EGCG can activate and up-regulate the expression of p16 gene mRNA which inhibits the proliferation of CA46 cell through inducing the G(0)/G(1) arrest by demethylation and/or by inhibiting DNMT3A and DNMT3B gene.

Synergistic effects of VPA and As2O3 on Molt-4 cells in vitro and its possible mechanisms / 中国实验血液学杂志

This study was purposed to investigate the synergistic effects of sodium valproate (VPA) and As2O3 on the proliferation of Molt-4 cells in vitro and its possible mechanisms. Cell viability and growth curve were assessed by the MTT assay. The synergistic activity in combination of 2 drugs was determined by the Q format. The expression levels of p15, DNA methyltransferase 1 (DNMT-1), DNMT3A and DNMT 3B mRNA were detected by RT-PCR and the methylation level was detected by hn-MSPCR. The results indicated that the VPA and As2O3 both inhibited proliferation of Molt-4 cells. The combination of two drugs showed an additive effect (values of Q were>or=0.85). The inhibitory rate in combination of 5 mmol/L of VPA with 10 micromol/L of As2O3 was (70.31+/-2.54)%. The p15 gene in Molt-4 cell line failed to express due to its hypermethylation. The level of p15 gene mRNA expression increased significantly after exposure to VPA in combination with As2O3 for 48 h. As compared with control group, the expression of DNMT-1 was down-regulated in a dose-dependent manner, whereas DNMT3A had no significant differences from the control. The level of expression of DNMT3B seemed to decrease at 10 mmol/L concentration. There were significant differences between the combination of the two drugs and the control group. The gray value of methylated bands decreased after the treatment of VPA alone and in combination with As2O3 in a dose-dependent manner. It is concluded that VPA induces demethylation of p15 INK4B gene by inhibiting the DNMT-1 and DNMT3B gene activities, which up-regulates the p15 gene, recovers its activity. The combination of VPA with As2O3 has the synergistic additive effect on the inhibition of cell viability, so that VPA can reduce the side effect of As2O3 on liver function, which would be verified in the clinical practice.

Detection of p16 gene methylation status in adult patients with acute leukemia by using n-MSP / 中国实验血液学杂志

The study was aimed to explore the relationship between patterns of methylation or deletion and the development of acute leukemia, and further to clarify the possible mechanism in the development of adult acute leukemia. Nested methylation-specific polymerase chain reaction (n-MSP) was adopted to analyze p16 gene methylation or deletion patterns in 82 adult acute leukemia patients with different subtypes and stages. The results indicated that rate of p16 gene methylation was 39.0% in 82 adult acute leukemia patients, among them, 41.4% in acute myelogenous leukemia (AML) and 33.3% in acute lymphoblastic leukemia (ALL). It were found that 36.6% of de novo AL patients and 54.5% of relapsed AL patients developed the hypermethylation of p16 gene. Out of the 82 patients, 6 seemed to have deletion of p16 gene, including 1 AML (1.7%) and 5 ALL (20.8%). There were no hypermethylation or deletion of p16 gene in the 16 controls. It is concluded that methylation of p16 gene may play a more important role than homozygous deletion of p16 gene in the leukemogenesis and progression of adult acute leukemia.

Detection of p15 gene methylation or deletion status in different malignant cell lines by using hemi-nested methylation specific polymerase chain reaction / 中国实验血液学杂志

This study was aimed to investigate the methylation or deletion status of p15 gene in different malignant cell lines, and further to clarify their roles in the development and progression of malignant tumors. Hemi-nested methylation specific polymerase chain reaction (hn-MSP) was adopted to analyze p15 gene methylation or deletion status in 20 malignant tumor cell lines and mononuclear cells or normal cell lines in healthy people, as well as to evaluate its sensitivity and specificity. The results showed that among all of the cell lines, Molt-4, KG1, NCE, Raji, SMMC-7221, CA46, SW480 and NCI-H446 were partial methylated with CDKN2B gene, and its sensitivity of detection of p15 gene methylation was up to 1.0 x 10(-5), also it had great specificity. Peripheral blood mononuclear cell (MNCs) from healthy volunteer, HL-60, HepG2, 293, HeLa, SGC7901, U266 and CEM were unmethylated; and K562, NB4, GMC, Jurkat seemed to have deletion or mutation of p15 gene. It is concluded that the incidence of p15 gene methylation or deletion in many tumours, especially malignant hematopathy, is frequent, they correlate with disease progression and prognosis. Hn-MSP is highly sensitive and specific in analyzing p15 gene methylation, deserving in clinical application.

Detection of promoter methylation of p16 gene in hematological malignant cell lines by nested methylation specific polymerase chain reaction / 中国实验血液学杂志

This study was aimed to investigate the efficiency of modified methylation-specific polymerase chain reaction i.e. nested methylation-specific polymerase chain reaction, used to detect the promoter methylation of p16 gene in six hematological malignant cell lines, and to explore the application in selection of hematological malignant cell lines with promoter hypermethylation, and make them to be an idel cell models for studying the relationship between gene methylation and expression. DNAs were denatured by NaOH and then were subjected to bisulfite modification and a nested-MSP was used to amplify the promoter region, nested MSP product of p16 gene promoter was analyzed and sequenced. The results showed that the hypermethylation of p16 gene was detected in CA46 and U266, however, Molt4, K562, HL-60 and Jurkat cell lines were unmethylated. In conclusion, p16 gene methylation in hematological malignant cell lines can be perfectly detected by nested-MSP method, which is simple, sensitive and specific for screening all kinds of hematological malignant cell lines with p16 gene methylated.