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1.
Acta Pharmaceutica Sinica ; (12): 46-63, 2022.
Article in Chinese | WPRIM | ID: wpr-913167

ABSTRACT

In recent years, immunotherapy has made great progress in clinical cancer therapy. However, the poor tumor specificity, low intra-tumoral penetration, and low cellular uptake in the systemic delivery of immunotherapeutic drugs lead to low efficacy and poor safety, limiting the development of immunotherapy. Active tumor-targeting nano drug delivery systems (aNDDS) can enhance the concentration of drugs in target cells through the interaction between surface-conjugated antibodies or ligands and the receptors on target cell membranes, providing a viable strategy for specific and efficient drug delivery. In addition, some specific types of cell membranes with the natural targeting ability have been exploited for the construction of biomimetic nanocarriers to improve the drug delivery efficiency. In view of the many advantages of active tumor-targeting nanocarriers, researchers also have designed a series of aNDDS for promoting antitumor immune responses and proved that they improved the efficacy and safety of immunotherapy. In this review, we summarize the recent progress on aNDDS for improving the tumor immunotherapy and look forward to the main challenges and future directions in this field.

2.
Article in Chinese | WPRIM | ID: wpr-905299

ABSTRACT

Objective:To observe the effects of pain relief after acupuncture on walking speed, step length and ground reaction force (GRF) of patients with chronic nonspecific low back pain (CNLBP) during walking. Methods:From May to December, 2019, 28 CNLBP patients were randomly divided into waiting list group (n = 14) and acupuncture group (n = 14). The acupuncture group received acupuncture, 30 minutes a time, three times a week, for four weeks. The waiting list group only received health education after enrollment until four weeks later. Gait analysis was performed with three-dimensional motion system for both groups after enrollment and one month later. The walking speed, step length and GRF characteristic values were recorded and compared, as well as Visual Analogue Score (VAS) for pain. Results:After intervention, The VAS decreased in both groups (t > 2.956, P < 0.05), and was lower in the acupuncture group than in the waiting list group (t = -2.844, P = 0.004). No significant difference in walking speed, step length and GRF characteristic values was found after intervention in both groups (P > 0.05). Conclusion:One month-acupuncture could relief the pain of CNLBP patients, however, it could not improve the performance during walking.

3.
Article in Chinese | WPRIM | ID: wpr-905393

ABSTRACT

Surface electromyography can record the neuroelectric signals responding the activity of muscle during exercise, which has been used to observe the characteristics of trunk and lower limb muscles in patients with chronic low back pain. It may be used in the researches about pathogenesis, diagnosis and treatment of chronic low back pain by comparing the surface electromyography signals between patients and normal controls during various activities.

4.
Article in Chinese | WPRIM | ID: wpr-844028

ABSTRACT

Objective: To compare the overall efficiency and cost of the construction and screening of libraries of Pseudomonas aeruginosa arylsulfatase (PAAS) through site-directed saturation mutagenesis with random primers (NNN) and precise primers for PCR. Methods: Site-directed mutagenesis libraries were constructed using the random primers (NNN) of the selected site, an equal-mole mixture of precise primers and a purified precise primer each time for the amplification of the fusion vector of Escherichia coli alkaline phosphatase (ECAP) and PAAS through PCR. The libraries were screened in a high-throughput mode through the assay of activity ratios of PAAS/mutants to ECAP after alkaline lysis of host cells. Three approaches for site-directed saturation mutagenesis were compared for their number of monoclones screened, the number of their expected mutants discovered, overall time consumed, overall cost and other pertinent factors. Results: The site-directed saturation mutagenesis of M72 with random primers for PCR resulted in only 10 among 20 expected mutants after the screening of over 600 monoclones, besides obvious codon bias. In contrast, no obvious codon bias was observed and there were already 18 among 20 expected mutants after the screening of less than 150 monoclones for site-directed saturation mutagenesis of G138 with similar random primers. The site-directed saturation mutagenesis of M72 with an equal-mole mixture of precise primers yielded all of the 19 expected mutants after the screening of less than 190 monoclones; the site-directed saturation mutagenesis of M72 with the purified precise primers for PCR one-by-one gave all of the 19 expected mutants after the screening of just 2 monoclones for every expected mutant. The use of the purified precise primers for PCR one-by-one was thus more favorable for the construction of libraries of site-directed saturation mutagenesis for the minimum number of monoclones screened, the least overall time and cost. In comparison to the use of the purified precise primers for PCR one-by-one, the use of random primers or the equal-mole mixture of precise primers for PCR to generate the libraries tolerated much greater overall cost and longer time. Conclusion: The generation and screening of the site-directed saturation mutagenesis libraries with precise primers for PCR one-by-one were more practical in elucidating the sequence-activity relationship while the use of equal-mole mixture of precise primers for PCR was preferable for the screening of positive mutants during site-directed saturation mutagenesis.

5.
Acta Pharmaceutica Sinica ; (12): 1802-1809, 2019.
Article in Chinese | WPRIM | ID: wpr-780306

ABSTRACT

Immunotherapy has emerged as one of the major modalities for clinical cancer therapy, along with surgery, chemotherapy, radiotherapy and targeted therapy. However, tumor-targeted delivery of immune therapeutics is challenged by a series of barriers including non-specific release, poor tumor penetration capacity, and insufficient cellular uptake of the therapeutic regimens, which seriously restricted the efficiency and efficacy of immunotherapy. To address above challenges, nanosized drug delivery systems (NDDS) have been extensively exploited to achieve tumor-targeted delivery of immunotherapy drugs. It has been well investigated that solid tumors are of unique characteristics including acidic, hypoxic and enzymatic extracellular microenvironment. Meanwhile, the tumor cells are of acidic, reductant and reactive oxygen species intracellular microenvironment. In recent years, a large variety of tumor microenvironment-activatable NDDS have been exploited to respond specifically to the stimulus of extracellular or intracellular tumor microenvironment for enhancing the accumulation, retention and penetration in the tumor tissue. These NDDS were also employed to promote intracellular uptake and tunable drug release inside the tumor cells. In this review article, we summarized the recent progress of our laboratory using the tumor microenvironment-activatable NDDS for immune efficient therapeutics delivery, and improved cancer immunotherapy. We also briefly discussed the challenges and provided perspective of NDDS-based cancer immunotherapy.

6.
Article in Chinese | WPRIM | ID: wpr-818980

ABSTRACT

Objective To examine the effect of low temperature on trehalose and trehalase levels in Culex pipiens pallens. Methods The fourth instar larvae and female adult mosquitoes of Cx. pipiens pallens were exposed at 4 ℃ for 0, 1, 3, 6, 12, 18, 24 h and 0, 1, 3, 6, 12, 18, 24, 36, 48, 72 h, respectively. Then, the trehalose and trehalase contents were detected by enzyme-linked immunosorbent assay (ELISA) in mosquitoes. Results The contents of trehalose and trehalase significantly increased in the larval and female adult mosquitoes post-exposure to low temperature. The changing trend of trehalose levels was consistent in the larval and female adult mosquitoes, and the highest levels were (2.458 8 ± 0.379 2) mg/g and (2.825 7 ± 0.211 1) mg/g 3 h post-exposure to low temperature, respectively. The trehalose and trehalase levels fluctuated greatly within the first 6 h post-exposure to low temperature. Following adaptation for a period of time, the trehalose and trehalase levels remained at a relatively high level. Conclusion Low temperature may induce the production of trehalose and trehalase in Cx. pipiens pallens, and the trehalose and trehalase may play an important role in the improvement of the cold resistance.

7.
Article in Chinese | WPRIM | ID: wpr-818528

ABSTRACT

Objective To examine the effect of low temperature on trehalose and trehalase levels in Culex pipiens pallens. Methods The fourth instar larvae and female adult mosquitoes of Cx. pipiens pallens were exposed at 4 ℃ for 0, 1, 3, 6, 12, 18, 24 h and 0, 1, 3, 6, 12, 18, 24, 36, 48, 72 h, respectively. Then, the trehalose and trehalase contents were detected by enzyme-linked immunosorbent assay (ELISA) in mosquitoes. Results The contents of trehalose and trehalase significantly increased in the larval and female adult mosquitoes post-exposure to low temperature. The changing trend of trehalose levels was consistent in the larval and female adult mosquitoes, and the highest levels were (2.458 8 ± 0.379 2) mg/g and (2.825 7 ± 0.211 1) mg/g 3 h post-exposure to low temperature, respectively. The trehalose and trehalase levels fluctuated greatly within the first 6 h post-exposure to low temperature. Following adaptation for a period of time, the trehalose and trehalase levels remained at a relatively high level. Conclusion Low temperature may induce the production of trehalose and trehalase in Cx. pipiens pallens, and the trehalose and trehalase may play an important role in the improvement of the cold resistance.

8.
Acta Pharmaceutica Sinica ; (12): 809-820, 2017.
Article in Chinese | WPRIM | ID: wpr-779663

ABSTRACT

This study was aimed to build a new photo-sensitive co-delivery liposomes which combine photodynamic therapy with chemotherapy to reverse drug resistance of breast cancer. Photodynamic photosen-sitizer chlorin e6 trimethyl ester (Ce6tM) and chemotherapeutic drug doxorubicin hydrochloride (DOX) were loaded into the liposomes (liposomes loaded with Ce6tM and DOX, CDL) by thin-film hydration extrusion and ammonium sulfate active loading methods. CDL was characterized with cryo-transmission electron microscopy (Cryo-TEM), dynamic light scattering particle size, zeta potentials and photo-sensitive DOX release behaviors in vitro. CDL cytotoxicity, singlet oxygen production, DOX accumulation, intracellular ATP level and cell cycle analysis in MCF7/ADR cells were evaluated. Finally, the tissue distribution of DOX and antitumor effects of CDL in BALB/c-nu nude mice bearing MCF7/ADR tumor were investigated. The results showed that the particle size of obtained CDL was 90.7 ± 1.1 nm and distributed uniformly. CDL possessed outstanding properties of photo-sensitive drug release profile. The accumulated release of DOX reached (96.52 ± 0.11)% in 2 min under 671 nm laser irradiation (2 W·cm-2). Interestingly, DOX in CDL could maintain rapid release after 671 nm laser irradiation with low power and short time (15 s, 0.25 W·cm-2). This phenomenon was caused by oxidation of unsaturated phospholipids in CDL under 671 nm laser irradiation and had nothing to do with the slightly elevated temperature. Photo-sensitive drug release behavior contributed to increased DOX accumulation in MCF7/ADR cells. The half inhibition concentration (IC50) of DOX in CDL laser group in MCF7/ADR cells was decreased by 601.9-fold compared with no laser group, which could be related to increased accumulation of DOX, decreased ATP levels and cell cycle arrest in MCF7/ADR cells. With the help of CDL, DOX accumulation in tumor was increased and in cardiac toxicity was reduced in vivo. CDL laser group showed a good anti-tumor effect. The tumor inhibition rate was (94.7 ± 6.2)%. These results suggest that CDL has a promising potential in reversing drug resistance of breast cancer.

9.
Article in English | WPRIM | ID: wpr-819474

ABSTRACT

OBJECTIVES@#To discuss the association between FGFR4 gene polymorphism rs351855 (Glu388Aly) and the susceptibility and chemotherapeutic effect of cervical cancer infected by high-risk type HPV.@*METHODS@#A total of 162 patients with high-risk HPV cervical cancer and 162 healthy women were collected and the genotypes of the FGFR4 rs351855 locus were detected. The genotype distributions in the two groups were compared. The cervical cancer patients were divided into four groups which namely good therapeutic effect group and bad therapeutic effect, recurrence or metastasis and no recurrence or metastasis group respectively, and the risks of different genotype on the curative effect and prognosis were analyzed by Logistic regression. The survival time of patients with different genotypes was compared.@*RESULTS@#There was no statistic difference in FGFR4 rs351855 genotype distribution between the patients group and control group (P > 0.05), among which the risk of chemotherapy failure on GA + AA patients was 3.257 times as much as that of the GG patients, and the risk of recurrence or metastasis of GA + AA patients was 2.783 times as much as that of the GG patients. For AA patients, the risk of chemotherapy failure and the risk of relapse and metastasis are 3.833 and 3.406 times, respectively, as much as that of the GG patients. The overall survival of GA and AA patients was shorter than that of the GG patients, and significant difference was found (χ = 7.098, P = 0.029). The difference in overall survival between GA + AA patients and GG patients was almost statistically significant (χ = 3.634, P = 0.057).@*CONCLUSIONS@#The FGFR4 rs351855 polymorphism is not associated with the susceptibility of high-risk HPV cervical cancer, but patients with gene A was at higher risk of unfavorable chemotherapy prognosis compared with patients with GG.

10.
Article in Chinese | WPRIM | ID: wpr-230065

ABSTRACT

To investigate the chemical constituents of ethyl acetate from Cirsium setosum, fifteen flavonoids were obtained by column chromatography on silica gel, MCI, Sephadex LH-20, and preparative HPLC. Their structures were identified as 4',5,6-trihydroxy-7-methoxyflavone(1), 4',5-dihydroxy-7,8-dimethoxyflavone(2), sorbifolin-6-O-β-glucopyranoside(3), kaempferol-7-O-α-L-rhamnoside(4), kaempferol(5), quercetin-3-O-β-D-glucosyl-7-O-α-L-rhamnoside(6), myricetin(7), myricetin-3-O-β-D-glucoside(8), 5,7- dihydroxy -3',4'- dimethoxyflavone(9), 3',4',5- trihydroxy-3,7-dimethoxyflavone(10), 3',3,4',5-tetrahydroxy-7-methoxyflavone(11), 3'-hydroxy-4',5,7-trimethoxyflavone(12), 7-hydroxy-3',4',5-trimethoxyflavone(13), 4',5-dihydroxy-2',3',7,8-tetramethoxylflavone(14), and 5-hydroxy-2',3',7,8-tetramethoxylflavone(15) by spectroscopic data analysis. All compounds were isolated from this plant for the first time.Compounds(1-15) were evaluated for their hypoglycemic activities by PTP1B enzyme model. Among them, compounds 2, 12, and 14 showed significant PTP1B inhibitory activities with IC₅₀ values of 2.54, 1.85, 2.11 μmol•L⁻¹, respectively.

11.
Journal of Experimental Hematology ; (6): 1009-1012, 2015.
Article in Chinese | WPRIM | ID: wpr-357229

ABSTRACT

<p><b>OBJECTIVE</b>This study was aimed to investigate the expression and clinical significance of Bmi-1 and P14 in extranodal NK/T-cell lymphoma (ENKTCL) tissue.</p><p><b>METHODS</b>Maxvision immunohistochemistry technique was used to detect the expression level of Bmi-1 and P14 in the tissues of 21 patients with ENKTCL and 11 normal lymph nodes. The correlation of Bmi-1 or P14 expression with the clinical features and the correlation between Bmi-1 and P14 expression were analyzed.</p><p><b>RESULTS</b>The expression of Bmi-1 protein was higher in tissues of ENKTCL than that in tissues of lymph nodes, and the Bmi-1 expression levels did not correlate with patients' sex, age, lactate dehydrogenase (LDH), International Prognostic Index (IPI) scores and B symptoms (P > 0.05), except for clinical stage (P < 0.05). The P14 protein expression level was lower in ENKTCL tissues than in normal lymph node tissues, which did not correlate with age, sex, LDH, IPI scores, clinical stage and B symptoms. Correlation test showed a negative correlation between Bmi-1 and P14 (r = -0.472, P = 0.031).</p><p><b>CONCLUSION</b>Bmi-1 protein over-expresses in ENKTCL tissues that may display a negative-regulation effect on P14 in the genesis and progress of ENKTCL.</p>


Subject(s)
Genes, Tumor Suppressor , Humans , Lymph Nodes , Lymphoma, Extranodal NK-T-Cell , Oncogene Proteins , Polycomb Repressive Complex 1
12.
Article in English | WPRIM | ID: wpr-820538

ABSTRACT

OBJECTIVE@#To investigate the expression and potential roles of G protein-coupled receptor kinase 6 (GRK6) in hepatocellular carcinoma (HCC) patients.@*METHODS@#Immunohistochemistry and Western blot was performed to determine GRK6 expression in 73 HCC samples. And the correlation with clinicopathological features was also analyzed.@*RESULTS@#GRK6 expression was significantly higher in HCC than that in normal hepatic tissue. GRK6 was positively correlated with proliferation marker Ki-67, clinical stage, metastasis and survival time.@*CONCLUSIONS@#Our results suggested that GRK6 overexpression plays an important role in HCC. Monitoring the expression of GRK6 maybe helpful in early diagnosis and prognosis of HCC.


Subject(s)
Adult , Aged , Biomarkers, Tumor , Metabolism , Blotting, Western , Carcinoma, Hepatocellular , Metabolism , Female , G-Protein-Coupled Receptor Kinases , Metabolism , Humans , Ki-67 Antigen , Metabolism , Liver Neoplasms , Metabolism , Male , Middle Aged , Prognosis , Risk Factors
13.
Chinese Journal of Stomatology ; (12): 664-668, 2013.
Article in Chinese | WPRIM | ID: wpr-274162

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the efficacy of flowable composite resin(FCR) as stress-absorbing liners in Class I cavity restorations in vitro.</p><p><b>METHODS</b>Thirty Class I cavities of 4 mm in diameter and 2 mm in depth were prepared in polycarbonate (PC) plates and divided into three groups, ten each. After application of an adhesive, cavities in each group were restored using one of the following methods: A: restored with Charisma without any lining of FCR; B: lined with Revolution Formula 2 twice before restoration with Charisma; C: lined with Teric Flow twice before restoration with Charisma. All cavities were observed under a photoelastic microscope and photoelastic images were recorded at 3 min and 24 h after curing and the shrinkage stresses on the cavity wall were calculated. The polymerization shrinkage(v%) of the three composite resins was measured using bonded discs method and their elastic moduli were measured according to ISO standard.</p><p><b>RESULTS</b>The shrinkage stresses at 3 min and 24 h of the three methods were as follows,A: (4.93 ± 0.28), (5.87 ± 0.40) MPa, B: (4.90 ± 0.30), (5.84 ± 0.33) MPa, and C: (4.76 ± 0.28),(5.83 ± 0.37) MPa.No significant difference was found in results among different groups. The polymerization shrinkage(v%) in group A,B, and C were (2.63 ± 0.04)%, (4.56 ± 0.06)%, and (3.98 ± 0.02)%. The elastic modulus in group A, B, and C were (9.59 ± 0.65), (4.25 ± 0.51), and (5.41 ± 0.79) GPa.</p><p><b>CONCLUSIONS</b>Under present study condition, using a FCR as stress-absorbing liner under composite resin restoration does not significantly decrease the polymerization shrinkage stresses at the cavity wall.</p>


Subject(s)
Composite Resins , Chemistry , Dental Cavity Lining , Dental Restoration, Permanent , Methods , Dental Stress Analysis , Materials Testing , Polymerization
14.
Chinese Medical Journal ; (24): 3246-3250, 2012.
Article in English | WPRIM | ID: wpr-316529

ABSTRACT

<p><b>BACKGROUND</b>Myelosuppression is the main dose-related toxicity of many chemotherapeutic drugs. The human multidrug resistance (mdr1) gene is well-known for its ability to confering drug resistance. In this study, we meant to transplant the placenta mesenchymal stem cells (P-MSCs) moderated by mdr1 gene into a nude mice model radiated by γ-Co(60) and to explore the chemoprotection for bone marrow (BM) toxicity.</p><p><b>METHODS</b>Human P-MSCs were isolated from trypsin-digested term placentas and then transduced by with reconstructed retroviral vector containing mdr1 gene and green fluorescent protein (GFP) reporter gene. The integration and expression of mdr1 gene was observed indirectedly by the expression of GFP. A nude mice model was constructed after irradiation with a sublethal dosage of γ-Co(60). These irradiated mice were transplanted with mdr1-MSCs through the caudal vein and then received paclitaxel (PAC) intraperitoneal chemotherapy. The Peripheral peripheral blood (PB) of the nude mice was collected, and the PB cells counts and values were determined using an automatic analyzer.</p><p><b>RESULTS</b>After PAC treatment, mdr1-MSCs transplanted mice showed markedly improved survival upon compared to MSCs transplanted mice (85.7% vs. 57.1%). White blood cell (WBC) and red blood cell (RBC) counts as well as the hemoglobin (Hb) values were significantly increased in PAC treated mdr1-MSCs mice compared to PAC treated control mice when PAC chemotherapy had been finished (all P < 0.05), but the difference was not found in the plateltes (PLT) count (P > 0.05).</p><p><b>CONCLUSION</b>Human P-MSCs moderated by mdr1 gene when transplanted into nude mice may provide chemoprotection for hematopoietic toxicity.</p>


Subject(s)
Animals , Bone Marrow , Cell Differentiation , Genetics , Physiology , Cells, Cultured , Erythrocytes , Metabolism , Female , Genes, MDR , Genetics , Physiology , Green Fluorescent Proteins , Genetics , Metabolism , Hemoglobins , Metabolism , Humans , Leukocytes , Metabolism , Mesenchymal Stem Cells , Cell Biology , Metabolism , Mice , Mice, Inbred BALB C , Mice, Nude , Placenta , Cell Biology , Pregnancy
15.
Article in Chinese | WPRIM | ID: wpr-343975

ABSTRACT

The present condition in the acupoint term translation was analyzed and its existent problems in this area were discussed in this paper. The authors suggested that in translating the terms of acupoints, the translation on the meaning of the acupoints should be added, in this way, it can not only keep the integrity in acupoint translation, but also make the inheritance of the Chinese precious culture of Traditional Chinese Medicine further available.


Subject(s)
Acupuncture Points , Animals , Culture , Humans , Medicine, Chinese Traditional , Translating
16.
Chinese Medical Journal ; (24): 800-805, 2008.
Article in English | WPRIM | ID: wpr-258588

ABSTRACT

<p><b>BACKGROUND</b>Most of gynecologic malignancies are sensitive to chemotherapy. Myelosuppression is the main dose-related toxicity of many chemotherapeutic drugs. The human multidrug resistance (mdr1) gene is well known for its ability to confer drug resistance. This study aimed to explore the feasibility of expression and resistance of mdr1 gene transduction into human placenta mesenchymal stem cells (P-MSCs) by retrovirus vector.</p><p><b>METHODS</b>Human P-MSCs were isolated from trypsin-digested term placentas, and their immunophenotypes and differentiation potential were evaluated. Human P-MSCs were transduced by reconstructed retroviral vector containing the mdr1 gene and green fluorescent protein (GFP) reporter gene. The integration and expression of the mdr1 gene were observed indirectly by the expression of GFP, and fluorescence-activated cell sorter was used to evaluate the functional activity of permeability glycoprotein (P-gp) encoded by the mdr1 gene. The stimulating test was made in vitro to show pleiotropic drug resistance of transfected cells.</p><p><b>RESULTS</b>The isolated, cultured and expanded P-MSCs expressed stem cell markers such as CD29, CD44 and CD73, and showed osteogenic and adipogenic differentiation potentials under appropriate conditions. The expression of P-gp in the non-transfected P-MSCs cells was (0.4 +/- 0.1)%, but increased to (28.1 +/- 4.7)% after gene transfection (P < 0.01). And positive staining of P-gp located mainly at cell membrane and cytoplasm. Accumulation and extrusion assays showed that P-gp expressed by the transfected cells had pump-functional activity and could efflux daunomycin out of cells. The analysis of cell survival confirmed that transfected P-MSCs had a characteristic of multidrug resistance with a significant increase in the resistance to anticancer agents.</p><p><b>CONCLUSIONS</b>Transfer and expression of human mdr1 gene mediated by retrovirus vector conferred P-MSCs drug resistance. It might provide a new alternative to chemoprotection strategies.</p>


Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1 , Genetics , Physiology , Cell Differentiation , Female , Genes, MDR , Humans , Immunophenotyping , Mesenchymal Stem Cells , Metabolism , Placenta , Cell Biology , Metabolism , Retroviridae , Genetics , Transfection
17.
Article in Chinese | WPRIM | ID: wpr-813589

ABSTRACT

OBJECTIVE@#To investigate the effects of DNA hypomethylation on mRNA and protein expression of perforin promotor in T cells.@*METHODS@#T cells were isolated from the peripheral venous blood of healthy donors by density gradient centrifugation. CD4(+) and CD8(+) subsets were isolated using Miltenyi beads and protocols provided by the manufacturer. Where indicated the T cells were stimulated with PHA for 24 h, then treated with 5-azaC for an additional 72 h. Genomic DNA, mRNA, and protein were isolated from untreated and 5-azaC-treated T cells. Purified DNA was treated with sodium bisulfite, the desired sequences were amplified in sequential fragments using nested PCR. The amplified fragments were cloned into bacteria DH5 alpha and 5 independent clones for each of the amplified fragments were sequenced. The expression of perforin was determined using real time RT-PCR and Western blot.@*RESULTS@#The perforin mRNA and protein in the CD4(+) and CD8(+) subsets treated with 5-azaC were significantly higher than those in the untreated subsets (P<0.05). The results of bisulfite genomic sequencing showed that the methylation of perforin promotor was significantly reduced in the treated cells compared with the untreated cells (P<0.05).@*CONCLUSION@#The mRNA and protein expression of perforin significantly increases in the CD4(+) and CD8(+) T cells treated with 5-azaC,which is associated with DNA hypomethylation of perforin promoter in T cells.


Subject(s)
Adult , Azacitidine , Pharmacology , Cells, Cultured , DNA Methylation , Humans , Perforin , Genetics , Promoter Regions, Genetic , Genetics , T-Lymphocyte Subsets , Metabolism
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