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1.
Article in Chinese | WPRIM | ID: wpr-878987

ABSTRACT

To investigate the effect of Gegen Qinlian Decoction(GQD) on enzyme activity, gene expression and methylation level of fatty acid synthase(FASN) in adipose tissue from rats with insulin resistance induced by high-fat diet. The 60% fat-powered high-fat diet was continuously given to male SD rats to induce the insulin resistance model. Then, they were divided into five groups randomly and administrated by gavage every day for 16 weeks with following drugs respectively: 10 mL·kg~(-1)water for control group(C) and insulin resistance model control group(IR), 1.65 g·kg~(-1)GQD per day for low-dose group(GQDL), 4.95 g·kg~(-1)GQD per day for medium-dose group(GQDM), 14.85 g·kg~(-1)GQD per day for high-dose group(GQDH), and 5 mg·kg~(-1) rosiglitazone per day for rosiglitazone group(RGN). Epididymal adipose tissue was taken to determine enzyme activity of FASN by colorimetric method, mRNA expression level of Fasn by quantitative Real-time PCR(Q-PCR) and CpGs methylation level between +313 and +582 by bisulfite sequencing PCR(BSP). These results showed that Fasn expression was significantly lowered in IR model rats compared with the control rats(P<0.01). Enzymatic activity and CpGs methylation level of Fasn in IR group showed downward trends. Low and medium-dose GQD can increase enzyme activity of FASN(P<0.05). Moreover, low-dose GQD increased the total CpGs methylation level of Fasn fragment between +313 and +582 in insulin resistance rats(P<0.05). For GQDM group, the methylation frequency of CpGs at positions +506 and +508(P<0.01) as well as the methylation frequency of CpGs on the binding sites of transcription factorzinc finger protein 161(P<0.05) were significantly increased. The methylation frequency of CpG at +442 position was positively correlated with Fasn expression(P<0.01, r=0.735), and methylation frequencies of CpGs at +345 and +366 positions were positively associated to enzyme activity of FASN respectively(P<0.05, r=0.479; P<0.01, r=0.640). In conclusion, GQD can reverse enzyme activity of FASN and methylation level of Fasn in adipose tissue of insulin resistant rats, and CpG sites at positions +506 and +508 may be the targets of GQD. The methylation level of CpGs at + 345 and + 366 sites were possibly related to FASN activity, while methylation of CpG at + 442 site may be closely correlated with mRNA level of Fasn. In addition, GQD did not significantly change mRNA expression level of Fasn, but effectively reversed enzymatic activity, suggesting that GQD may regulate the post transcriptional expression of Fasn.


Subject(s)
Adipose Tissue , Animals , Drugs, Chinese Herbal , Fatty Acid Synthases/genetics , Gene Expression , Insulin Resistance/genetics , Male , Methylation , Rats , Rats, Sprague-Dawley
2.
Article in English | WPRIM | ID: wpr-880576

ABSTRACT

OBJECTIVE@#To evaluate the safety and effectiveness of Qishe Pill () on neck pain in real-world clinical practice.@*METHODS@#A multi-center, prospective, observational surveillance in 8 hospitals across Shanghai was conducted. During patients receiving 4-week Qishe Pill medication, Visual Analogue Scale (VAS) and Neck Disability Index (NDI) assessments have been used to assess their pain and function, while safety monitoring have been observed after 2 and 4 weeks.@*RESULTS@#Results from 2,023 patients (mean age 54.5 years) suggest that the drug exposure per unit of body mass was estimated at 3.41 ± 0.62 g/kg. About 8.5% (172/2,023) of all participants experienced adverse events (AEs), while 3.8% (78/2,023) of all participants experienced adverse reaction. The most common AEs were gastrointestinal events and respiratory events. The VAS score (pain) and NDI score (function) significantly decreased after 4-week treatment. An effect-quantitative analysis was also conducted to show that the normal clinical dosage may be consider as 3-4 g/kg, at which dosage the satisfactory pain-relief effect may achieve by 40-mm reduction in VAS.@*CONCLUSION@#These findings showed that patients with cervical radiculopathy who received Qishe Pill experienced significant improvement on pain and function. (Registration No. NCT01875562).

3.
National Journal of Andrology ; (12): 589-597, 2017.
Article in Chinese | WPRIM | ID: wpr-812911

ABSTRACT

Objective@#To investigate the effects of down-regulation of PTTG1 expression on the proliferation, invasiveness and apoptosis of androgen-independent human prostate cancer LNCaP-AI cells and their sensitivity to androgen antagonists.@*METHODS@#Human prostate cancer LNCaP-AI cells were transfected with siRNA targeting the PTTG1 gene using the Lipofectamine 2000 transfection reagent. The proliferation, invasiveness and apoptosis of the cells were detected by MTT, Transwell assay and flow cytometry, respectively. The protein expressions of PTTG1, p-Akt, and p-ERK were determined by Western blot and the mRNA expression of PTTG1 measured by agarose gel electrophoresis.@*RESULTS@#The siRNA expression vector markedly down-regulated the expression of PTTG1, which effectively suppressed the proliferation of the LNCaP-AI cells, with the inhibition rates of (19.47 ± 2.12), (24.01 ± 2.13) and (48.02 ± 2.22)% at 24, 48 and 72 hours, respectively, after transfection, with statistically significant differences among the three groups (P <0.05). The number of the cells passing through the polycarbonate film was remarkably decreased at 24, 48 and 72 hours (74.67 ± 9.85, 56.44 ± 8.66 and 37.33 ± 6.14) as compared with the baseline (111.11 ± 13.47) (P <0.01), while the apoptosis rate of the cells was significantly increased at 24, 48 and 72 hours (18.32 ± 0.94), (19.94 ± 1.30) and (21.73 ± 1.88)% in comparison with the baseline ([2.17 ± 0.49]%), (P <0.05). PTTG1 siRNA combined with androgen antagonist flumatide exhibited even more significant effects in inhibiting the proliferation and promoting the apoptosis of the LNCaP-AI cells than either used alone, and in a flumatide dose-dependent manner. The inhibition and apoptosis rates of the LNCaP-AI cells treated with 50 nmol/L flumatide were (27.13 ± 3.52) and (3.94 ± 0.48)%, and those treated with siRNA + 50 nmol/L flumatide were (67.51 ± 5.13) and (19.93 ± 1.72)%, respectively, both with statistically significant differences between the two groups (P <0.05). The inhibition and apoptosis rates of the cells treated with 100 nmol/L flumatide were (43.72 ± 3.90) and (5.33 ± 0.66)%, and those treated with siRNA + 100 nmol/L flumatide were (73.19 ± 4.78) and (23.43 ± 1.76)%, respectively, both with statistically significant differences between the two groups (P <0.05).@*CONCLUSIONS@#The siRNA expression vector can down-regulate the expression of PTTG1, which can inhibit the proliferation and invasiveness of LNCaP-AI cells, promote their apoptosis, and increase their sensibility to androgen antagonists. Suppressing the expression of PTTG1 may enhance the effect of androgen-deprivation therapy on advanced prostate cancer.


Subject(s)
Androgen Antagonists , Pharmacology , Apoptosis , Cell Line, Tumor , Cell Proliferation , Down-Regulation , Humans , Male , Neoplasm Invasiveness , Prostatic Neoplasms , Drug Therapy , Metabolism , Pathology , RNA, Small Interfering , Metabolism , Securin , Genetics , Metabolism , Time Factors , Transfection
4.
Chinese Pharmaceutical Journal ; (24): 373-378, 2016.
Article in Chinese | WPRIM | ID: wpr-859189

ABSTRACT

OBJECTIVE: To study the antitumor activity of TWS119 and its effect on the killing activity of γδT cells in vitro. METHODS: CCK-8 assay was used for detecting the influence of TWS119 on the proliferation of SGC-7901 and γδT cells and the cytotoxic activity of γδT cells to SGC-7901 cells. The cell apoptosis was measured by flow cytometric analysis. The protein expression was examined by Western blot analysis. RESULTS: It was found that TWS119 could inhibit the growth and induce the apoptosis of SGC-7901 cells. The expression of Bcl-2 in SGC-7901 cells treated with TWS119 was downregulated, while p-ERK1/2 and p-STAT3 were up-regulated. In addition, TWS119 at 0.5-4.0μmol · L-1 could promote the growth of γδT cells and enhance the cytotoxic activity of γδT cells to SGC-7901 cells especially at 4.0 μmol · L-1 for 72 h. Furthermore, TWS119 could upregulate the expression of Bcl-2 and inhibit the expression of p-ERK1/2 and p-STAT3. CONCLUSION: TWS119 in some concentrations can inhibit the growth of SGC-7901 cells, promote the 78T cells growth and enhance the cytotoxic activity of γδT cells to SGC-7901 cells, and the mechanism may be involve in Wnt, ERK1/2, Bcl-2 and p-STAT3 signaling pathways.

5.
National Journal of Andrology ; (12): 686-691, 2016.
Article in Chinese | WPRIM | ID: wpr-262324

ABSTRACT

<p><b>Objective</b>To explore the expression of pituitary tumor transforming gene 1 (PTTG1) during the transformation of prostate cancer from androgen-dependent (ADPC) to androgen-independent (AIPC).</p><p><b>METHODS</b>We established an AIPC cell model LNCaP-AI by culturing the androgen-dependent LNCaP cell line in the hormone-deprived medium for over 3 months. The cell model was verified and the PTTG1 expression in the LNCaP cells was detected by Western blot and RT-PCR during hormone deprivation.</p><p><b>RESULTS</b>The AIPC cell model LNCaP-AI was successfully established. The PTTG1 expression was gradually increased in the LNCaP cells with the prolonged time of hormone deprivation and the expressions of matrix metalloproteinases MMP-2 and -9 were elevated at the same time.</p><p><b>CONCLUSIONS</b>The expression of PTTG1 is increased gradually in AIPC, which may be a target of gene therapy for advanced prostate cancer.</p>


Subject(s)
Blotting, Western , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , Humans , Male , Matrix Metalloproteinase 2 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Neoplasms, Hormone-Dependent , Prostatic Neoplasms , Genetics , Securin , Genetics
6.
Article in Chinese | WPRIM | ID: wpr-258139

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effects of Nourishing yin clearing heat ([Chinese characters: see text]) Chinese herbal medicine on vascular endothelial growth factor (VEGF) in growth retardation induced by Decamethasone and observe its mechanisms.</p><p><b>METHODS</b>Thirty one-month-old New Zealand white rabbits were randomly divided into normal group, dexamethasone-treated group and Nourishing yin clearing heat ([Chinese characters: see text]) Chinese herbal medicine-treated group. The rabbits in dexamethasone group and Nourishing yin clearing heat ([Chinese characters: see text]) Chinese herbal medicine-treated group received dexamethasone (5 mg/kg x d). The rabbits were sacrificed at the 6th and 12th week after administration, and then rabbit tibia articular was removed. (1) Using TUNEL stain to detect apoptotic index. (2) Using immunohistochemical stain to detect the positive index of the expression of vascular endothelial growth factor (VEGF) in the epiphyseal cartilage of growth. (3) Using fluorescent quantitative PCR to detect the expression intensity of VEGF mRNA in each group.</p><p><b>RESULTS</b>At the 6th and 12th week after administration, there were significant difference in apoptotic index and cell proliferation index between dexamethasone group and normal group (P<0.01, dexamethasone group more than normal group). Immunohistochemical stain and fluorescent quantitative PCR indicated that the expression of VEGF and VEGF mRNA in dexamethasone group was significantly decreased as compared with that in normal group (P<0.01), and also obviously lower than Chinese herbal medicine-treated group (P<0.01).</p><p><b>CONCLUSION</b>VEGF has 2. an important role during the growth retardation induced by Dexamethasone. Nourishing yin clearing heat ([Chinese characters: see text]) Chinese herbal medicine can reduce the growth retardation induced by Dexamethasone through increasing the VEGF expression in growth plate chondrocytes and then increase angiogenesis.</p>


Subject(s)
Animals , Apoptosis , Cell Proliferation , Dexamethasone , Pharmacology , Drugs, Chinese Herbal , Pharmacology , Female , Gene Expression , Growth Plate , Cell Biology , Metabolism , Male , Rabbits , Random Allocation , Vascular Endothelial Growth Factor A , Genetics , Metabolism
7.
Article in Chinese | WPRIM | ID: wpr-316179

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression of connective tissue growth factor (CTGF) in osteoarthritis chondrocytes, and the relationship between CTGF, macrophage colony-stimulating factor (M-CSF) and cartilage degeneration.</p><p><b>METHODS</b>Ten New Zealand white rabbits were randomly divided into the normal group and OA model group established by Hulth's method. Sections were stained with Safranin O for histological examination. The cartilage histological characteristics were observed according to the method of Mankin. Immunohistochemical staining was performed. Articular cartilages were observed with microscopy and the image analysis method was used to measure the expression intensity of CTGF and M-CSF in each group, and the correlations of the expression of CTGF, M-CSF and cartilage degeneration were analyzed by statistics.</p><p><b>RESULTS</b>Immunohistochemical staining indicated that the expression intensity of CTGF, M-CSF in untreated group was significantly increased as compared with that in the normal group (P<0.05). Statistical analysis showed that there was a correlation between the expression of CTGF, M-CSF and cartilage degeneration (r=0.634, r=0.542, P<0.01 respectively).</p><p><b>CONCLUSION</b>The expression of CTGF and M-CSF protein is up regulated in osteoarthritis chondrocytes, which suggests that the activation of M-CSF is involved in the production of CTGF. CTGF and M-CSF play an important role in the pathogenesis of cartilage degeneration.</p>


Subject(s)
Animals , Cartilage , Pathology , Chondrocytes , Chemistry , Pathology , Connective Tissue Growth Factor , Female , Immunohistochemistry , Macrophage Colony-Stimulating Factor , Male , Osteoarthritis , Metabolism , Pathology , Rabbits
8.
Article in Chinese | WPRIM | ID: wpr-263723

ABSTRACT

<p><b>OBJECTIVE</b>To explore the influence of Yanghe decoction on the expression of HIF-1alpha mRNA in osteoarthritis (OA), so as to study mechanisms of Yanghe decoction in the improvement of cartilagedegeneration.</p><p><b>METHODS</b>Thirty New Zealand white rabbits were randomly divided into 3 groups: control group (10 rabbits), OA model group (10 rabbits) and Yanghe decoction treatment group (10 rabbits). The OA models were established by using Hulth method. The sections were stained with Safranin O for histological examination. Each sample was evaluated the cartilages histological characteristics according to the method of Mankin. Fluorescent quantitative PCR was used to detect the expression of HIF-1alpha mRNA in each group.</p><p><b>RESULTS</b>There were significant differences in the Mankin score between the control group and the OA model group (P = 0.005), either between the OA model group and the Yanghe decoction treatment group (P = 0.003). Fluorescent quantitative PCR indicated that HIF-1alpha mRNA significantly increased in the OA model group compared to that of the control group (P = 0.035), but that expression was much lower in Yanghe decoction treatment group compared to that of the OA model group (P = 0.039).</p><p><b>CONCLUSION</b>The expression of HIF-1alpha mRNA plays an important role in OA. Yanghe decoction is effective to protect the articular cartilage, which is achieved through regulating the expression of HIF-1alpha mRNA.</p>


Subject(s)
Animals , Drugs, Chinese Herbal , Therapeutic Uses , Female , Hypoxia-Inducible Factor 1, alpha Subunit , Genetics , Male , Osteoarthritis , Drug Therapy , Metabolism , RNA, Messenger , Rabbits
9.
Article in Chinese | WPRIM | ID: wpr-343953

ABSTRACT

Activating blood circulation to remove stasis method is an important therapy of TCM, which can be matched with various methods, as qi-supplementing, qi-regulating, heat-clearing with detoxication, meridian warming, wind-dispelling to remove dampness, yin-nourishing, phlegm-dissolving to alleviate depression and visceral dredging by purgation, to produce various effects on angiogenesis. Its positive or negative impacts on quality or quantity of angio-genetic regulatory factor play a crucial part for the effects. This article explores the effect and mechanism of activating blood circulation to remove stasis method on angiogenesis bi-directionally.


Subject(s)
Angiogenic Proteins , Metabolism , Blood Circulation , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Hematologic Agents , Pharmacology , Therapeutic Uses , Hematologic Diseases , Drug Therapy , Humans , Medicine, Chinese Traditional , Qi
10.
Microbiology ; (12)1992.
Article in Chinese | WPRIM | ID: wpr-686263

ABSTRACT

The reform and practice on teaching approaches of microbiology integrated case-based study,heuristic teaching,interaction teaching,comparison expression teaching and inductive learning was explored,which was contraposed the education aims of independent college and student's trait,and combined to the fact of University of Electronic Science and Technology of China Zhongshan Institute.The result of practice showed that student enthusiasm for study was mobilized,and better results in teaching were got by applica-tion of diversiform teaching approaches.

11.
Microbiology ; (12)1992.
Article in Chinese | WPRIM | ID: wpr-684032

ABSTRACT

By using enrichment media NPC and CVP, 792 strains of Pseudomonas and 515 strains of Erwinia were isolated from the rhizosphere of Digitaria adscendens (H. B. K.) Hem and Setaria vindis (L.) Beanv. Following which, experiments of antimetabolic test with E. coli, seed emergence controlling of S. viridis, herbicidal activity and security with green grass were carried out to select the desired bacteria. As the result, the selected strain, S7, could wholly control the seed emergence of S. viridis without any harm to the two tested green grass. And more, S7 promoted the seed emergence of Festuca arundinacea slightly. In spite of the comparatively low corrected mortality (56. 7%) of S7 after emergence of S. viridis, Selecting of microbial herbicides from weed DRB is thought to be more prospective.

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