Study on shedding particles from Chuanping Sustained-release Tablets based on multi-marker components balanced release / 中草药

Objective: To study on the changes of shedding particles, drug loading and release of Chuanping Sustained-release Tablets (CST) which were made by the different sustained-release excipients in vitro, so as to elucidate the mechanism about balanced release of multi-marker components on Chinese materia medica compound sustained-release preparation. Methods: Using Chuanping Prescription (Ephedra Herba and Datura Flos) as model drug, improved Peach Gum and HPMC was used as sustained release materials, the release test combinied with underwater video observation were applied to shot external forms at different time point, and the laser particle sizer was applied to determinate particle size, and HPLC was applied to determine the accumulated release rate of the index components (ephedrine, pseudoephedrine, and scopolamine) to calculate the cumulative release curve slope K value, and to evaluate the balance release of the different components. Results: CST which were made by the improved peach gum and HPMC sustained-release excipient, which particles were dropped at 0.5 h after contacting the dissolution medium. At the same time, the particles were obviously shedding with the passage of time. In contrast, CST that was made by the improved peach gum, its balanced release of multi-marker components was better (K of ephedrine was 12.18, K of pseudoephedrine was 12.30, and K of scopolamine was 12.40), and particles dropped faster (it was significantly at 1 h), and particle size was bigger (D50 was 53.37—70.33 μm and D90 was 100.3—196.5 μm), and drug loading was more (ephedrine 30.63%, pseudoephedrine 32.97%, and scopolamine 31.67%), and release time of drug was longer (60—120 min). Conclusion: The shedding particles were important part of balance release of multi-marker components about CST which was made by the improved peach gum sustained-release excipient, and also was the embodiment of the drug release mode of “corrosion-dissolution”.

Changes of polysaccharide content, functional properties and in vitro release of peach gum before and after improvement / 中草药

Objective To analyze the difference of polysaccharides content, property characterization, and in vitro release between the original peach gum and improved peach gum, and to find the change rule, so as to provide the basis for its further application as the new delivery material. Methods The content of polysaccharides was measured by the phenol-sulfuric acid method combined with ultraviolet spectrophotometry. The pH value of liquid cement was measured by pH meter. The dynamic viscosity of liquid cement was measured by the Brookfield DV-Ⅱ Pro viscometer. The solubility and swelling ratio of rubber powder were measured by the weighing method. The moisture-absorption rate at different relative humidity of rubber powder was measured by the method of controlling humidity with the desiccator. With three index components (ephedrine, pseudoephedrine, and scopolamine) as the standards, HPLC and the release test were applied to determine the accumulated release rate of Chuanping adhesive tablets (CAT). Results Original peach gum: In the three producing areas of Xishui in Guizhou Province, Xinfeng in Jiangxi Province, and Suixian in Hubei Province, the content of polysaccharides respectively was 84.30%, 81.52%, and 77.84%. The pH value of 1% liquid cement was 5.63, 5.60, and 5.88. The dynamic viscosity of 1% liquid cement was 30.8, 28.5, and 25.9 mPa∙s. The solubility of rubber powder was 6.08%, 5.75%, and 5.12%. The swelling ratio of rubber powder was 34.22%, 29.66%, and 26.25%. The 12 h moisture-absorption rate of the improved peach gum polysaccharose at RH of 43% was 28.19%, 27.02%, and 26.44%; and at RH of 81% was 38.22%, 35.26%, and 34.64%. The 2 h cumulative release amount in vitro of methamphetamine, pseudoephedrine, and scopolamine of Chuanping adhesive tablets were 95.41%—98.84%, 96.05%—97.11%, and 96.35%—98.21%. Improved peach gum: In the three producing areas of Xishui, Xinfeng, and Suixian, the content of polysaccharides respectively was 96.92%, 94.76%, and 93.35%. The pH of 1% liquid cement was 7.10, 7.08, and 7.12. The dynamic viscosity of 1% liquid cement was 318.4, 289.6, and 266.4 mPa∙s. The solubility of rubber powder was 17.82, 16.73, and 16.38 g. The swelling ratio of rubber powder was 84.98%, 81.55%, and 79.82%. The 12 h moisture-absorption rate of the improved peach gum polysaccharose at RH of 43% was 41.22%, 39.93%, and 39.20%; and at RH of 81% was 60.88%, 58.48%, and 57.17%. The 12 h cumulative release amount in vitro of methamphetamine, pseudoephedrine, and scopolamine of CAT were 97.88%—98.36%, 97.59%—98.56%, and 97.72%—98.12%. Conclusion Compared with the original peach each gum, the content of polysaccharides, dynamic viscosity, solubility, swelling ratio, and moisture-absorption rate, and sustained release property of the improved peach gum were all improved remarkably, and the solution was also neutral. These results show that the improved peach gum can be further developed and applied as a new sustained release material.

Study on balanced release of Chuanping extract sustained-release tablets combined with different accessories / 中草药

Objective: To study the balanced release of complex components of Chuanping extract sustained-release tablets combined with different accessories. Methods: With methamphetamine, pseudoephedrine, and scopolamine as indexes, HPLC and the in vitro release test were applied to determining the accumulated release rate of the complex components in nine sustained-release Chuanping extract preparations (Q1-Q9), and to evaluating the influences of different accessories to the release behaviors of complex components in vitro. Results: The differences about dissolutions of four single modern materials in Chuanping extract sustained-release tablets Q1-Q3 were significant, and Q4 released fast. All of them were not balanced release. The differences about dissolutions of single improved Chinese materia medica (CMM) sustained-release accessories made Chuanping extract sustained-release tablets Q5 were significant, and Q6 was balanced release. The difference about dissolutions of complex improved CMM sustained release excipients made Chuanping extract sustained-release tablets Q9 was incomplete, the balanced release of Q8 was not well. The balanced-release of complex components of Q7 was well. Conclusion: Different accessories would affect the balanced release of sustained-released Chuanping extract tablet in vitro. The modern sustained-release accessories are hard to achieve the balanced release of complex components of CMM compound sustained-release preparations, but the improved traditional sustained-release accessories could be achieved well.

Correlation between dissolution in vitro and absorption in vivo of chuanping sustained release tablets / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the correlation between dissolution in vitro and absorption in vivo of Chuanping sustained release tablets.</p><p><b>METHOD</b>The ephedrine, pseudoephedrine were chosen as marker components, dissolution in vitro of Chuanping sustained release tablets in the different pH were tested by the rotating basket method and HPLC; urine drug levels were determined by HPLC and absorption fractions were calculated according to Wagner-Nelson's formula and deconvolution technique.</p><p><b>RESULT</b>The linear regressive equation between the absorption percentage in vivo F and accumulative release percentage in vitro of Chuanping sustained release tablets was established as F(ephedrine) = 1.572 5f-20. 729 (R2 = 0.974 5); F(pseudoephedrine) = 1.237f-0.147 6 (R2 = 0.959 5).</p><p><b>CONCLUSION</b>The results suggested that there was fine correlation between the absorption percentage in vivo and the accumulative release percentage in vitro of Chuanping sustained release tablets.</p>

Evaluation of balanced release of complex components of Chuanping sustained tablets based on fingerprint / 中国中药杂志

To combine fingerprint and drug release rate in vitro, in order to study in vitro release of complex components of Chuanping sustained tablets, compound traditional Chinese medicine preparation. A qualitative determination of the characteristic peaks of the compound preparations were conducted by the fingerprint. The results of the dissolution rate determination under different release conditions showed that the release of three index components (methamphetamine, pseudoephedrine and scopolamine) of Chuanping sustained tablets was less affected by gastrointestinal factors, with similarity factors being more than 80 with unknown component release curves of three major characteristic peaks in the fingerprint. The qualitative determination proved that multiple components of the compound traditional Chinese medicine preparation was dissolved in vitro at similar rates, realizing the balanced release of complex components of the compound traditional Chinese medicine preparation. This study layed a theoretical and experimental basis for quality evaluation for the compound traditional Chinese medicine preparation.

Demethylation of the gamma-synuclein gene CpG island in colorectal cancer and its clinical significance / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the relationship between gamma-synuclein gene expression and CpG island demethylation in colorectal cancer(CRC), and the relationship between the demethylation and clinicopathological factors of CRC.</p><p><b>METHODS</b>The expression of gamma-synuclein mRNA was examined in 30 pairs of tumor tissues and tumor-matched non-neoplastic adjacent tissues(NNAT) by RT-PCR. CRC cell lines including COLO205, LoVo, and SW480 were used and treated with a demethylating agent, 5-aza-2'-deoxycytidine(5-aza-C). Before and after the treatment, the expression of gamma-synuclein mRNA in the cells was determined by RT-PCR, and bisulfite sequencing PCR was also used to analyze methylation status of CpG island. The methylation status of gamma-synuclein was then examined in 67 CRC samples and 30 NNAT samples by nested methylation-specific PCR (NMSP) and real time methylation-specific PCR(real-time MSP). The relationship between the demethylation of gamma-synuclein in CRC and clinicopathological factors was analyzed.</p><p><b>RESULTS</b>The mean gamma-synuclein mRNA expression was 0.66+/-0.34 in CRC samples, which was much higher than 0.45+/-0.26 in NNAT samples(P=0.011). 5-aza-C could induce expression and demethylation of gamma-synuclein in COLO205, LoVo and SW480 cells. gamma-Synuclein gene was demethylated in 80.0%(24/30) of the CRC samples and 50.0%(15/30) of the NNAT samples. The demethylated status of gamma-synuclein was much higher in CRC samples than that in NNAT samples(P=0.030), and was significantly correlated with clinical stage, lymph node involvement, and distant metastasis of CRC(P<0.05).</p><p><b>CONCLUSION</b>The upregulation of gamma-synuclein expression in CRC is primarily attributed to the demethylation of CpG island, which may be used as a marker for prognosis.</p>

Hedgehog signaling pathway activates in gastric carcinoma and promotes the proliferation through GLI1 in MKN28 cell / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of Hedgehog (HH) pathway on proliferation and in vitro tumorigenicity of gastric cancer cell lines.</p><p><b>METHODS</b>The expression of SHH, PTCH, SMO, SUFU and GLI1 in seven cell lines were tested by RT-PCR. siRNA targeting GLI1 mRNA was transfected into MKN28 cells. Cell proliferation and in vitro tumorigenicity were examined by CCK8 and soft agar colony formation test.</p><p><b>RESULTS</b>SHH in six gastric cancer cell lines was up-regulated. Expression of PTCH in KATOIII cell lines and expression of SUFU in MKN28 and KATOIII were reduced. GLI1 siRNA significantly inhibited the expression of GLI1 in MKN28 cell line. Growth rate and colony formation rate of MKN28 cells treated with GLI1 siRNA were significantly lower than those of control cells (all P <0.001).</p><p><b>CONCLUSIONS</b>HH signaling pathway is widely activated in gastric cancer cell lines. The activation of HH signaling pathway promotes the growth of MKN28 cells.</p>

Expression and intracellular localization of FRZB gene in gastric cancer and its significance / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To study the expression and intracellular localization of FRZB gene in gastric cancer tissue, and to explore its significance in gastric cancer.</p><p><b>METHODS</b>The expression of FRZB in tumor tissues from 90 patients with gastric cancer and in normal gastric mucous as control were analyzed by immunohistochemistry in tissue array. FRZB expression in gastric cancer cell lines and immortalized gastric epithelial cell line GES-1 were detected by quantitative real-time PCR(Q-PCR) and Western blot. The intracellular localization of FRZB was observed by immunofluorescence staining.</p><p><b>RESULTS</b>The positive expression rate of FRZB in gastric cancer was 92.2%. FRZB expressed in gastric cancer with well differentiation was higher than that with poor differentiation.The positive rate in normal gastric mucous was 10.0% (one out of ten). By confocal microscope, FRZB localized both in cytoplasma and nucleus, especially on the nuclear membrane. The Q-PCR and Western blot results also showed that the expression of FRZB in gastric cancer cell lines was higher than that in GES-1.</p><p><b>CONCLUSIONS</b>The expression of FRZB in gastric cancer is correlated with tumor cell differentiation and tumor Lauren classification. The nuclear localization of FRZB may contribute to its function in gastric cancer formation and progression.</p>

Polo like kinase 1 expression and prognostic value in gastric carcinomas / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To identify the expression of polo like kinase 1 (plk1) and to discuss its relationship with the clinicopathological parameters and prognosis in gastric carcinoma.</p><p><b>METHODS</b>Plk1 protein expression levels in 89 cases of resected gastric carcinomas were detected by immunohistochemistry method, the relations between plk1 expression levels and the survival periods were estimated by Kaplan-Meier curve.</p><p><b>RESULTS</b>The positive rate of plk1 expression in gastric cancer tissues was 42.7% (38/89), significantly higher than that (13.5%) in the adjacent noncancerous tissues (12/89) (P<0.01). The expression levels of plk1 were closely related to tumor differentiation, invasion and TNM stage (P<0.05). Patients with plk1-positive expression had worse prognosis than those with plk1-negative expression in gastric cancer patients (P<0.05).</p><p><b>CONCLUSIONS</b>Plk1 may promote carcinogenesis and gastric cancer development, its overexpression can be a novel marker for diagnosing certain biological behaviours and predicting prognosis in gastric cancer.</p>

Mitotic arrest of gastric cancer cells induced by silencing of STK15 gene / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the role of STK15 in regulating mitosis of gastric cancer cells (MKN45) by gene silencing through RNA interference mechanism.</p><p><b>METHODS</b>RNA interference technique was used to inhibit STK15 expression in MKN45 cells. The expression levels of STK15 mRNA and protein were measured by real-time quantitative RT-PCR and Western blot respectively and cell morphological changes were investigated by reverse microscopy. In addition, cell cycle distribution and cellular proliferation were determined by flow-cytometry and MTT assay respectively. Finally, the mitotic phenotype of MKN45 cells was studied by immunofluorescence staining and confocal microscopy.</p><p><b>RESULTS</b>Silencing of STK15 gene by RNA interference was confirmed by marked decrease of STK15 mRNA and protein levels in the treated MKN45 cells. This silencing correlated with rounding of the cells, decreasing of DNA content in G(2) phase (P < 0.05) and a lowered proliferation index (P < 0.05), along with alterations of mitotic phenotype of MKN45 (P < 0.05).</p><p><b>CONCLUSION</b>STK15 gene may play a key role in regulating cellular mitosis and its inhibition by RNA interference leading to mitosis arrest in MKN45 cells.</p>

Expression and single nucleotide polymorphisms of kallikrein 10 in colorectal cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To demonstrate expression and single nucleotide polymorphisms (SNP) of human kallikrein 10 (KLK 10) in colorectal cancer (CRC) and to correlate the KLK 10 expression level with clinicopathological factors of CRC.</p><p><b>METHODS</b>KLK 10 expression in 63 cases of tumoral and nontumoral colorectal tissues at the mRNA and protein levels were evaluated by quantitative real-time RT-PCR (qRT) and Western blot methods. KLK 10 protein was localized by immunohistochemistry. The KLK 10 genomic DNA from 16 cases of paired normal and cancerous colorectal tissues was PCR-amplified and examined for SNP by direct sequencing.</p><p><b>RESULTS</b>The KLK 10 mRNA expression was detected by qRT in 61 of 63 (97%) CRC specimens. The KLK 10 expression was much higher in tumor tissue than in the corresponding normal mucosal tissue at the mRNA and protein levels. The KLK 10 mRNA expression level significantly correlated with the lymphatic invasion (P < 0.05) and clinical stage of CRC (P < 0.05). No mutations or polymorphisms were detected in exon 1, 2 and 5 of KLK 10 gene in CRC. A SNP in codon 50 of exon 3, GCC (alanine) to TCC (serine) was identified. The genetic changes of exon 4 were located at codon 106 [GGC (glycine) to GGA (glycine)], codon 112 [ACG (threonine) to ACC (threonine)], codon 141 [CTA (leucine) to CTG (leucine)], and codon 149 [CCG (proline) to CTG (leucine)]. All these SNP were identical in tumor as well as the corresponding normal tissue DNA from the same individuals.</p><p><b>CONCLUSIONS</b>The KLK 10 expression is up-regulated in CRC and higher expression of KLK 10 closely correlate with advanced disease stage, which predicts a poorer prognosis, however, further follow-up study is needed.</p>

Study of growth inhibition of gastric cancer cells by sRNA targeting polo like kinase 1 in vitro and vivo / 中华外科杂志

<p><b>OBJECTIVE</b>To observe the effect of polo like kinase 1 (plk1) gene depletion on the growth of gastric cancer cell line-MKN45 cells in vitro and vivo and discuss the feasibility and effectiveness of arranging plk1 as gene therapeutic target for gastric cancer.</p><p><b>METHODS</b>The plk1 expression of MKN45 cells was inhibited by RNA interference (RNAi). The plk1 mRNA and protein level were measured by real-time quantitative PCR and western blotting, and the change of cell cycle distribution and apoptosis rate were detected by flow-cytometry, and the MKN45 cells proliferation was measured by MTT method. MKN45 cells treated with plk1 siRNA were transplanted subcutaneously in nude mice and their tumorgenesis ability were observed, the plk1 protein levels of the samples from nude mice in different groups were compared.</p><p><b>RESULTS</b>After treatment with plk1 siRNA, plk1 mRNA and protein level decreased obviously in certain time, more MKN45 cells accumulated at G(2)/M (P < 0.05). Apoptosis rate of MKN45 cells treated with plk1 siRNA was higher than that of control cells at 48 h and 72 h (P < 0.05), and MKN45 cells proliferated slowly than control groups (P < 0.05), while the tumorgenesis ability obviously decreased, but the plk1 protein levels of the samples from nude mice in different groups were not different.</p><p><b>CONCLUSIONS</b>siRNA targeting plk1 can inhibit the proliferation of MKN45 cells in vitro and vivo. Plk1 may be a novel therapeutic target for gastric cancer.</p>

Mitosis arrest caused by inhibition of PLK1 expression in gastric cancer MKN45 cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To observe the effect of polo-like kinase 1 (PLK1) gene depletion on mitosis phenotype and elucidate its vital role in gastric cancer cell line (MKN45) mitosis.</p><p><b>METHODS</b>The PLK1 expression in MKN45 cells was blocked by RNA interference (RNAi), the expression level of PLK1 mRNA and protein were measured by real-time quantitative PCR and Western blot, respectively. The morphological change of microtubules and mitosis phenotype in MKN45 cells were observed by immunofluorescence staining and laser confocal microscopy, the morphological changes of cells were observed by reverse microscopy, the variation of cell cycle distribution was detected by flow-cytometry.</p><p><b>RESULTS</b>After RNAi targeting PLK1, PLK1 mRNA and protein level decreased obviously, the cell microtubules became obscure and lost cohesiveness, the mitosis phenotype also varied substantially (P < 0.05), more gastric cancer cells became rounded and showed G(2) phase cell DNA content (P < 0.05).</p><p><b>CONCLUSION</b>PLK1 gene plays a key role in mitosis and its inhibition can lead to mitosis arrest in MKN45 cells.</p>

Inhibition of polo like kinase gene expression induces apoptosis in gastric cancer cells / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To observe the effect of inhibition of polo like kinase1 (plk1) gene expression on apoptosis induction in gastric cancer cell line-MKN45 and discuss the vital role of plk1 proliferation and viability of gastric cancer cells.</p><p><b>METHODS</b>The plk1 expression was inhibited by chemically synthesized siRNA. The plk1 mRNA and protein level were respectively measured by real-time quantitative PCR and Western blotting. The spindle morphological change was observed by immunofluorescence staining and confocal microscopy. The change of cell cycle distribution and apoptosis rate was detected by flow-cytometry. Pro caspase3 level was also detected by western blotting.</p><p><b>RESULTS</b>After treatment by siRNA targeting plk1, plk1 mRNA and protein level decreased obviously, the cell mitotic spindle became obscure and lost cohesiveness, more MKN45 cells accumulated at G(2)/M phase (P< 0.05), apoptosis rate of plk1 siRNA treated MKN45 cells was higher than that of control cells at 48 h and 72 h (P< 0.05) with pro-caspase3 level decreasing at 72 h.</p><p><b>CONCLUSIONS</b>Inhibition of plk1 gene expression induces apoptosis in MKN45 cells through the pathway of caspase3. Plk1 gene play a key role in viability of MKN45 cells.</p>

Identification of biomarkers in the serum of the patients with poorly differentiated gastric cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the application of proteomics in the field of serology,and to screen the differential expression proteins related with poorly differentiated gastric carcinoma.</p><p><b>METHODS</b>Two-dimensional electrophoresis (2-DE) was applied to segregate the total proteins in the serum form gastric cancer patients and health volunteers. After staining,the differential expression proteins were analyzed using PDQuest software,and identified using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS).</p><p><b>RESULTS</b>Electrophoresis figures with high resolution and reproducibility were obtained. Six differential expression proteins were found only in the serum from gastric cancer patients, while four other proteins from healthy volunteers.</p><p><b>CONCLUSIONS</b>Protein expression is differential in the serum from the gastric cancer patients and health volunteers. It is hopeful to find the biomarkers related with poorly differentiated gastric carcinoma using proteomics.</p>

Inhibition of serine/threonine kinase 15 gene expression induces apoptosis of gastric cancer cells / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To observe the effect of inhibition of serine/threonine kinase15 (STK15) gene expression on apoptosis induction in gastric cancer cell line-MKN45 and discuss the role of STK15 in viability of gastric cancer cells.</p><p><b>METHODS</b>The STK15 expression was inhibited by chemically synthesized siRNA. The STK15 mRNA and protein level were respectively measured by real-time quantitative PCR and western blotting,the change of cell cycle distribution and apoptosis rate were detected by flow-cytometry, cell morphological change was observed by Hoechst staining,and pro-caspase 3 level was also detected by western blot.</p><p><b>RESULTS</b>After treatment by siRNA targeting STK15 after 48 h, STK15 mRNA and protein level decreased obviously. More MKN45 cells accumulated at G(2)/M phase (P< 0.05). The apoptosis rate of STK15 siRNA treated MKN45 cells was higher than that of control cells(P< 0.05) with the pro-caspase 3 level decreased.</p><p><b>CONCLUSIONS</b>Inhibition of STK15 gene expression may induce apoptosis in MKN45 cells through the pathway of caspase3. STK15 gene play a key role in proliferation and viability of MKN45 cells.</p>

Expression of liver receptor homolog 1 gene in patients with cholesterol gallstone disease / 中华消化杂志

Objective To investigate the mRNA expression of liver receptor homolog 1 (LRH-1) gene in patients with cholesterol gallstone disease so that to elucidate the biomolecular pathogenesis of gallstone for- mation.Methods Twenty-seven patients with cholesterol gallstone (CGS) and 14 controls were included in this study.Biliary composition was assayed and mRNA expression of hepatic LRH 1 gene was determined by real time polymorphism chain reaction.Results In CGS patients,expression of LRH-1 was significantly higher than that in controls (14.18?9.37 vs 7.86?6.19,P＜0.05),and cholesterol of bile was oversaturated (1.17?0.27).Conclusion The formation of CGS may be related to increased expression of hepatic LRH-1 gene.

Inhibition of Plk-1 gene expression enhences 5-Fu and CDDP induced apoptosis of gastric cancer cells / 中国普通外科杂志

Objective To observe the effect of inhibition of polo like kinasel (Plk-1) gene expression on 5-Fu and CDDP induced apoptosis of gastric cancer cell line-MKN45 in vitro. Methods The Plk-1 expression was inhibited by chemically synthesized siRNA, the Plk-1 mRNA and protein level were measured by real-time PCR and Western blotting, MKN45 cells proliferation was measured by MTT method, the apoptosis rate was detected by flow-cytometry. Results After treatment by siRNA targeting Plk-1, Plk-1 mRNA level decreased by 51.91% .89.45%.91.03% at 24 h.48 h and 72 h, and Plk-1 protein level decreased by 38.43% and 60.66% at 48 h and 72 h, compared with control group. The proliferation of MKN45 cells treated by a combination of 5-Fu, CDDP and Plk-1 siRNA significantly slowed down when compared with treatment of 5-Fu, CDDP or Plk-1 siRNA(P

The expression of DAO in gastric cancer cells and gastric mucosa cells / 肠外与肠内营养

Objective:To study the expression of D-amino acid oxidase(DAO) in gastric cancer cells and gastric mucosa cells. Methods:The expression of DAO was detected in seven gastric cancer cell lines and a normal gastric mucosa cell line GES-1 by real-time quantitative PCR and in tumor tissues and normal gastric mucous tissues of 46 patients with gastric cancer by RT-PCR.The liver and kidney tissues of nude mouse acted as positive controls.Results:DAO was expressed in the liver and kidney tissues of nude mouse,and DAO expression in kidney tissue was higher than that in liver tissue.DAO was not detected in seven gastric cancer cell lines and the normal gastric mucosa cell line GES-1.Except one tumor tissue sample,DAO was not detected in other gastric cancer tissues and all normal mucosa tissues.Conclusion:DAO was not expressed in gastric cancer cells and gastric mucosa cells.The nutritional treatment of D-Met in place of L-Met could avoid the disadvantage of methionine starvation to important organs,such as liver and kidney.

In vitro and in vivo growth inhibition of gastric cancer cells by siRNA targeting STK15 / 中国普通外科杂志

Objective To observe the effect of STK15 gene silencing on the growth of gastric cancer cell line-MKN45 in vitro and vivo. Methods STK15 expression was inhibited by RNAi techenique, STK15 protein level was detected by Western blot, the ability of MKN45 invasion in vitro was assessed by cell migration and invasion assay, the change of cell cycle distribution was detected by flowcytometry, MKN45 proliferation was measured by MTT method, and MKN45 cells treated with STK15 siRNA were transplanted subcutanuously in nude mice and their tumorgenesis ability were observed. Results After treatment with STK15 siRNA, STK15 protein level decreased obviously. Compared with control group, STK15- group showed lower invasion ability in vitro [ mean A value: (182?27 ) vs. (308?38 ), P