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.

A new diarylheptanoid from barks of mangifera indica / 中草药·英文版

Objective: To study the chemical constituents from the barks of Mangifera indica. Methods: The constituents were separated and purified by different methods of chromatography, and their structures were elucidated by IR, MS, 1D and 2D NMR techniques. Results: Six compounds were isolated from the barks of M. indica. Their structures were identified as mangiferone (1), mangiferin (2), myricetin (3), myricitrin (4), rutin (5), and quercetin (6). Conclusion: Mangiferone (1) is a new diarylheptanoid compound isolated from the barks of M. indica. © 2013 Tianjin Press of Chinese Herbal Medicines.

Inhibition of malignant phenotype of HeLa cells by RNA interference of the telomerase activity / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the biology of HeLa cells upon inhibition of human telomerase catalytic subunit (hTERT) gene by RNA interference in vitro.</p><p><b>METHODS</b>Four shRNAs (A, B, C and D) targeting hTERT gene were designed and prepared by in-vitro transcription. The expression of hTERT gene was evaluated by immunofluorescent staining and telomeric repeat amplification protocol (TRAP) ELISA (TRAP-ELISA), after transient transfection of shRNAs by lipid formulation. Through the initial selection, shRNA (B) was noticed as the most efficient one in down-regulating hTERT gene and therefore was chosen as the ultimate shRNA used in the experimental groups. Those transfected by non-silencing RNAi were chosen as the control groups. Cell spreading and migration were studied by microscopy and cell adhesion to fibronectin (FN) was assayed by cell counting kit-8 (CCK-8). Cell invasion was assessed by Boyden chamber assay.</p><p><b>RESULTS</b>Cell spreading study revealed that the rates of spreading cells in the experimental groups were (5.6 +/- 2.3)% at 30 min, and (26.3 +/- 6.1)% 2 h after the inoculation, respectively, whereas the rates of spreading cells in the control groups were (31.3 +/- 7.9)% and (79.4 +/- 4.8)%, respectively. There were significant differences between the two groups (P < 0.01). However, most of the cells in both groups became spreading after 24 h. Cell adhesion assay demonstrated that the rate of adhesion cells on FN in experimental groups was (67.2 +/- 2.8)%, less than that in control groups (83.7 +/- 5.4)% (P < 0.05). The relative migration distance was (27.1 +/- 6.2)% in the experimental group, lower than that of the control group (58.7 +/- 15.0)%. The invasion assay revealed that the invading cells were 75.7 +/- 14.5 in the experimental group, in contrast to 165.1 +/- 11.0 in the control group after 4 h incubation on matrigel. The difference between these two groups was significant (P < 0.05).</p><p><b>CONCLUSION</b>In vitro shRNA silencing of hTERT gene can down-regulate the telomerase activity, leading to an inhibition of the malignant phenotype of HeLa cells, including decreased ability of cell spreading and adhesion, reduction of cell migration, and declined invasive ability through Matrige assay.</p>

Telomerase activity,PI3K/AKT signaling pathway and cellular biological behavior in HeLa cell line / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the telomerase activity and to document biological behaviors of HeLa cells upon treatment with specific PI3K/AKT signaling pathway inhibitor, LY294002.</p><p><b>METHODS</b>CCK-8 assay was used to determine IC50 of LY294002. The expressions of total AKT and phosphorylation AKT (P-AKT) were determined using Western blot. Telomerase activity of cell was measured by TRAP-ELISA assay. Cell growth curve, flow cytometry technique and Hoechst33258 stain were used to evaluate the cell growth, cell cycle and apoptosis respectively. Cell migration was determined by cell wound healing assay.</p><p><b>RESULTS</b>IC50 value of LY294002 of treated HeLa cells was 1.73 mg/L. Western blot showed that LY294002 enabled to decrease P-AKT activity in the presence of same total AKT protein. The cell telomerase activity was decreased to 36.72% in contrast to 98.61% of the control. LY294002 decreased the telomerase activity in HeLa cells, and the growth capacity of the cells was significantly suppressed. The number of cells at G0/G1 phases increased to 66.88% compared with that of the control cells (47.36%). The apoptosis rate also increased from 2.4% to 14.9%. The relative migration distance decreased to 24.6% compared with that of control (62.57%).</p><p><b>CONCLUSION</b>LY294002 inhibition of PI3K/AKT signaling pathway leads to alteration of telomerase activity along with changes of the biological behaviors of the HeLa cells suggesting that regulation of telomerase activity may be closely related to PI3K/AKT signaling pathway.</p>

Biologic characteristics of rat bone marrow mesenchymal stem cells cultured in vitro / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate biological characteristics of rat bone marrow mesenchymal stem cells (MSC) cultured in vitro and to explore their potential applications.</p><p><b>METHODS</b>MSC were isolated from rat bone marrow by density gradient centrifugation and were induced to differentiation. Flow cytometry was used to characterize their surface antigen expression, cell cycle status and cell growth parameters. Telomerase activity was determined by TRAP-ELISA assay.</p><p><b>RESULTS</b>Fusiform MSC became larger and flattener with increasing passages of culture. After the fourth passage, the MSC showed an immunophenotype of CD29 (94.75% +/- 3.68%), CD71 (95.43% +/- 2.23%), and CD90 (98.08% +/- 3.88%). After the seventh passage, MSC with such immunophenotype decreased with CD29: 50.00% +/- 3.35%, CD71: 50.70% +/- 2.43%, and CD90: 48.60% +/- 2.83%. Cells with such immunoprofile completely disappeared after passage 9. Overall, MSC grew faster during the first 5 passages. The number of MSC in S and G(2)/M phases were 38.36% +/- 2.01% and those in G(0)/G(1) phase were 61.64% +/- 2.13% after 3 passages. The cell growth decreased after passage 7. Percentage of MSCs in S and G(2)/M phases was 10.83% +/- 1.63% and that in G(0)/G(1) was 89.17% +/- 1.96% after passage 12, after which the cells failed to further divide. After passage 9, MSCs lost their ability to differentiate to Von Kossa and oil red O positive staining cells. In addition, telomerase activity of MSC also gradually decreased with the prolonged passages, from the original 52.7% +/- 0.78% to no telomerase activity.</p><p><b>CONCLUSION</b>The biological and immunophenotypical characteristics of cultured MSC showed obvious alterations with increasing numbers of passage of culture.</p>