Clinical significance of CEA mRNA expression in peritoneal lavage fluid for patients with gastric cancer after radical surgery / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the expression and clinical significance of CEAmRNAin peritoneal lavage fluid for patients with gastric cancer after radical surgery. Methods: The clinical data of 139 gastric cancer patients, who underwent peritoneal lavage CEA mRNA detection after radical resection in the Comprehensive Cancer Centre of Drum Tower Hospital from January 2013 to December 2017 were retrospectively analyzed. Routine post-operative follow-up was conducted in all patients. The expression of CEA mRNA in peritoneal lavage fluid after radical resection of 139 gastric cancer patients was detected by RT-PCR. Chi-square test analysis was used to study the relationship between the expression of CEA mRNA in peritoneal lavage fluid and basic clinical features, histopathological data, hematological indicators and the recurrence pattern of GC patients. Logistic univariate and multivariate regression analyses were used to screen the influential factors affecting CEA mRNA expression. Results: CEA mRNA was positive in 44 (31.7%) of 139 patients. Analysis showed that there was no significant correlation between CEA mRNA expression and sex, age, pathological grade, Lauren type, HER2, EGFR, VEGFR and Ki67 (all P>0.05), but there was significant correlation between CEA mRNA expression and pathological type, vascular invasion, local invasion depth, lymph node metastasis and clinical AJCC stage (all P<0.05). The peritoneal recurrence rate of patients with positive CEA mRNA expression was significantly higher than that of patients with negative expression (P=0.012). Logistic univariate regression analysis showed that signet ring cell carcinoma (P=0.04, HR=2.810， 95% CI: 1.050-7.520), T stage (P=0.016，HR=6.329, 95% CI: 1.417-28.264), N stage (P=0.022，HR=3.068，95% CI: 1.172-8.027), AJCC stage (P=0.016，HR= 3.971, 95% CI: 1.295-12.173), nerve invasion (P=0.002, HR=6.738, 95% CI: 1.995-22.757) and vascular invasion (P<0.001, HR= 16.36, 95% CI: 3.85-69.512) were risk factors for positive CEA mRNA expression in peritoneal lavage fluid of patients with gastric cancer. Logistic multivariate regression analysis showed that vascular invasion (P<0.001, HR=21.314，95% CI: 4.21-107.907) was an independent risk factor for positive CEAexpression in peritoneal lavage fluid of gastric cancer patients. Conclusion: Gastric cancer patients with positive CEA mRNA in peritoneal lavage fluid have higher risk of peritoneal recurrence or metastasis and poorer prognosis. So, more aggressive anti-tumor treatments including local abdominal cavity treatment should be considered.

Sipi soup inhibits cancer­associated fibroblast activation and the inflammatory process by downregulating long non­coding RNA HIPK1­AS.

Sipi soup (SPS), the aqueous extract derived from the root bark of Sophora japonical L, Salix babylonica L., Morus alba L., as well as Amygdalus davidiana (Carr.) C. de Vos, is a traditional Chinese medicine frequently used to prevent and treat infection and inflammation. However, the role of SPS in cancer­associated fibroblasts (CAFs) require further investigation. In the present study, the effects of SPS on fibroblast inactivation and the underlying mechanism were investigated. Reverse transcription­quantitative polymerase chain reaction was used to analyze the mRNA expression levels of fibroblast activation protein (FAP), interleukin (IL)­6, α­smooth muscle actin (α­SMA) and programmed cell death 4 (PDCD4). Flow cytometry was used to evaluate cell apoptosis. Immunofluorescence was used to determine the number of activated fibroblasts. The present study reported that SPS treatment did not affect the proliferative apoptotic potential of fibroblasts. Treatment with HeLa cell culture medium (CM) induced a significant increase in the expression levels of FAP, IL­6 and α­SMA, but reduced the expression of PDCD4. SPS reversed the effects of HeLa CM on the expression of these genes. Analysis with a long non­coding (lnc)RNA array of numerous differentially expressed lncRNAs revealed that the expression levels of the lncRNA homeodomain­interacting protein kinase 1 antisense RNA (HIPK1­AS) were increased in cervicitis tissues and cervical squamous cell carcinoma tissues compared with in normal cervical tissues. HIPK1­AS expression levels were upregulated in response to HeLa CM, but were decreased under SPS treatment. The downregulation of HIPK1­AS expression via short hairpin RNA abolished the effects of HeLa CM on the expression of inflammation­associated genes. The findings of the present study suggested that SPS may prevent the progression of cervical cancer by inhibiting the activation of CAF and the inflammatory process by reducing HIPK1­AS expression.

Dietary Lycopene Supplementation Improves Cognitive Performances in Tau Transgenic Mice Expressing P301L Mutation via Inhibiting Oxidative Stress and Tau Hyperphosphorylation.

BACKGROUND: Oxidative stress is implicated in the pathogenesis of Alzheimer's disease (AD) and other tauopathies and participates in their development by promoting hyperphosphorylation of microtubule-associated protein tau. Lycopene, as an effective antioxidant, combined with vitamin E seemed to be additive against oxidative stress. OBJECTIVE: The present study was undertaken to examine whether lycopene or lycopene/vitamin E could exert protective effects on memory deficit and oxidative stress in tau transgenic mice expressing P301L mutation. MATERIALS AND METHODS: P301L transgenic mice were assigned to three groups: P301L group (P301L), P301L+lycopene (Lyc), and P301L+lycopene/vitamin E (Lyc+VE). Age-matched C57BL/6J mice as wild type controls (Con) were used in the present study. Spatial memory was assessed by radial arm while passive memories were evaluated by step-down and step-through tests. Levels of tau phosphorylation were detected by western blot. Oxidative stress biomarkers were measured in the serum using biochemical assay kits. RESULTS: Compared with the control group, P301L mice displayed significant spatial and passive memory impairments, elevated malondialdehyde (MDA) levels and decreased glutathione peroxidase (GSH-Px) activities in serum, and increased tau phosphorylation at Thr231/Ser235, Ser262, and Ser396 in brain. Supplementations of lycopene or lycopene/vitamin E could significantly ameliorate the memory deficits, observably decreased MDA concentrations and increased GSH-Px activities, and markedly attenuated tau hyperphosphorylation at multiple AD-related sites. CONCLUSIONS: Our findings indicated that the combination of lycopene and vitamin E antioxidants acted in a synergistic fashion to bring significant effects against oxidative stress in tauopathies.

Multi-Vitamin B Supplementation Reverses Hypoxia-Induced Tau Hyperphosphorylation and Improves Memory Function in Adult Mice.

Hypobaric hypoxia (HH) leads to reduced oxygen delivery to brain. It could trigger cognitive dysfunction and increase the risk of dementia including Alzheimer's disease (AD). The present study was undertaken in order to examine whether B vitamins (B6, B12, folate, and choline) could exert protective effects on hypoxia-induced memory deficit and AD related molecular events in mice. Adult male Kunming mice were assigned to five groups: normoxic control, hypoxic model (HH), hypoxia+vitamin B6/B12/folate (HB), hypoxia+choline (HC), hypoxia+vitamin B6/B12/folate+choline (HBC). Mice in the hypoxia, HB, HC, and HBC groups were exposed to hypobaric hypoxia for 8 h/day for 28 days in a decompression chamber mimicking 5500 meters of high altitude. Spatial and passive memories were assessed by radial arm and step-through passive test, respectively. Levels of tau and glycogen synthase kinase (GSK)-3ß phosphorylation were detected by western blot. Homocysteine (Hcy) concentrations were determined using enzymatic cycling assay. Mice in the HH group exhibited significant spatial working and passive memory impairment, increased tau phosphorylation at Thr181, Ser262, Ser202/Thr205, and Ser396 in the cortex and hippocampus, and elevated Hcy levels compared with controls. Concomitantly, the levels of Ser9-phosphorylated GSK-3ß were significantly decreased in brain after hypoxic treatment. Supplementations of vitamin B6/B12/folate+choline could significantly ameliorate the hypoxia-induced memory deficits, observably decreased Hcy concentrations in serum, and markedly attenuated tau hyperphosphorylation at multiple AD-related sites through upregulating inhibitory Ser9-phosphorylated GSK-3ß. Our finding give further insight into combined neuroprotective effects of vitamin B6, B12, folate, and choline on brain against hypoxia.

[Effect of eletroacupuncture for Z syndrome without organic lesion and serum leptin].

OBJECTIVE: To explore the effect and mechanism of electroacupuncture (EA) for Z syndrome without organic lesion (metabolic syndrome combined with obstructive sleep apnea hypopnea syndrome). METHODS: Fifty-eight patients with Z syndrome were divided into three groups according to mild,moderate and severe degree. Acupuncture and EA were used at Daimai (GB 26), Zhongwan (CV 12), Xiawan (CV 10), Zusanli ST 36), Qihai (CV 6) and Huaroumen (ST 24), etc., once a day and five times a week. The treatment of ten times was a course, and two courses were acquired continuously. Sleep respiration monitoring (PSG) was done before EA and in one week after treatment respectively. Triacylglycerol (TG) fasting blood glucose (FBG) fasting insulin (INS) and serum leptin (Lep) were tested before and after treatment in the three groups. RESULTS: After treatment apnea hypopnea index (AHI) and the percentage of the time of arterial oxygen saturation (SaO2) less than 90% taken in the total sleep time (SLT 90%) were improved apparently than those before treatment in the three groups (all P < 0.05). The levels of TG, FPG, INS and Lep were decreased after treatment in all groups (all P < 0.05). CONCLUSION: EA can improve AHI and nocturnal hypoxia of Z syndrome, and the mechanism may be related to decreasing the indices of metabolism syndrome and leptin.

DHA in Pregnant and Lactating Women from Coastland, Lakeland, and Inland Areas of China: Results of a DHA Evaluation in Women (DEW) Study.

Few studies have examined docosahexaenoic acid (DHA) in pregnant and lactating women in developing countries like China, where DHA-enriched supplements are increasingly popular. We aimed to assess the DHA status among Chinese pregnant and lactating women residing areas differing in the availability of aquatic products. In total, 1211 women in mid-pregnancy (17 ± 2 weeks), late pregnancy (39 ± 2 weeks), or lactation (42 ± 7 days) were enrolled from Weihai (coastland), Yueyang (lakeland), and Baotou (inland) city, with approximately 135 women in each participant group by region. DHA concentrations were measured using capillary gas chromatography, and are reported as weight percent of total fatty acids. Mean plasma DHA concentrations were higher in coastland (mid-pregnancy 3.19%, late pregnancy 2.54%, lactation 2.24%) and lakeland women (2.45%, 1.95%, 2.26%) than inland women (2.25%, 1.67%, 1.68%) (p values < 0.001). Similar differences were observed for erythrocyte DHA. We conclude that DHA concentrations of Chinese pregnant and lactating women are higher in coastland and lakeland regions than in inland areas. DHA status in the study population appears to be stronger than populations from other countries studied to date.

[HPLC specific chromatograms of drug pair of schizonepetae herba and saposhnikoviae radix].

In this experiment, the HPLC specific chromatogram was adopted, with Agilent Extend-C18 (4.6 mm x 250 mm, 5 microm) as the chromatographic column, and 0.5 per thousand trifluoroacetic acid and acetonitrile as the mobile phase for gradient elution, so as to establish specific chromatograms for drug pair of Schizonepetae Herba and Saposhnikoviae Radix from different producing area, identify 12 common characteristic peaks, and obtain the comparison specific chromatography of drug pair of Schizonepetae Herba and Saposhnikoviae Radix. The method is simple, accurate and highly reproducible, and thus can be used as the basis for the quality control of the drug pair.

[Mixing behavior of nutrients in different seasons at Liaohe Estuary].

The distribution and species of nutrients in the Daliaohe Estuary were studied in April, July and November, 2010. Concentrations of nutrients displayed obviously seasonal variability in the Daliaohe Estuary. Highest concentrations of PO4(3-) -P, DOP, DTP and TP were in April, and the lowest were in July. While the highest PP was in November, the lowest was in July. The contents of NO3(-) -N and NO2(-) -N reached the highest point in July, and the lowest was in April; however, the highest NH4(+) -N, DON and PN were in April and the lowest were in November. The main existing form of TN was NH4(+) -N in April, while in July and November NO3(-) -N was the dominant. The behavior of NO3(-) -N,DIN and TN was conservative in various seasons; the behavior of NO2(-) -N, NH4(+) -N and DON was conservative in April and November, while nonconservative in July, which indicated the exist of the extra source in the interior of the estuary. The SiO3(2-) of the Daliaohe Estuary was conservative, with the highest average concentration in November and the lowest in April.

Synergistic anticancer effects of polyphyllin I and evodiamine on freshly-removed human gastric tumors.

OBJECTIVE: The present study was designed to examine the anticancer effect of Traditional Chinese Medicine of polyphyllin I (PPI) and evodiamine (EVO) on freshly-removed gastric tumor tissues. METHODS: Sixty freshly-removed gastric tumor tissues were collected. Their sensitivity to PPI, EVO, platinum (Pt), 5-FU, irinotecan (CPT-11) were determined by histoculture drug response assay (HDRA). Those samples were also formalin-fixed and paraffin-embedded, which were used to examine the mRNA expression levels of aprataxin(APTX), excision repair cross-complementing 1(ERCC1), thymidylate synthase(TS) and topoisomerase I(TOPO1) by quantitative RT-PCR. The association of the gene expression levels and in vitro sensitivity were analyzed. RESULTS: PPI, EVO, Pt, 5-FU and CPT-11 had anticancer effects on the freshly-removed gastric tumor tissues with average inhibition rates of 20.64%±14.25% for PPI, 21.14%±13.43% for EVO, 50.57%±22.37% for Pt, 53.54%±22.03% for 5-FU, and 39.33%±24.79% for CPT-11, respectively. Combination of PPI and Pt, EVO and Pt, EVO and 5-FU had higher inhibition rates than any single drug of them (P<0.001, P = 0.028, P = 0.017, respectively). The mRNA expression levels of ERCC1 were correlated with Pt sensitivity (rho = -0.645, P<0.001); the mRNA expression levels of TS were correlated with 5-FU sensitivity (rho = -0.803, P<0.001). There were also weak but significant correlations between APTX mRNA expression levels and CPT-11 sensitivity (rho = -0.376, P = 0.017) or EVO sensitivity (rho = -0.322, P = 0.036). ERCC1 mRNA expression levels was markedly suppressed by the presentation of PPI (P = 0.001) and slightly suppressed by the presentation of EVO (P = 0.04); whereas, TS mRNA expression levels was markedly decreased by the presentation of EVO (P = 0.017) and slightly decreased by the presentation of PPI (P = 0.047). CONCLUSION: PPI and EVO both could inhibit the activity of freshly-removed gastric tumor, and they could enhance the anticancer effect of Pt and 5-FU by reducing the mRNA expression levels of ERCC1 and TS.

Rhizoma Paridis ethanol extract selectively inhibits the proliferation of HUVECs comparing to Lovo cells and shows anti-angiogenesis effects in a mouse model.

ETHNOPHARMACOLOGICAL RELEVANCE: Rhizoma Paridis, a traditional Chinese Medicine, was identified to be cytotoxic to cancer cells. The present study was designed to investigate the potential anti-angiogenic and antitumor effect of the ethanol extract of Rhizoma Paridis (RPE) in vitro and in vivo. MATERIALS AND METHODS: The cytotoxic effect of RPE against human colon cancer Lovo cells and human umbilical vein endothelial cells (HUVECs) was examined using MTT assay. We also tested the effect of RPE on tube formation, migration, apoptosis and cell cycle of HUVECs. Moreover, Lovo subcutaneous xenograft was applied to study the antitumor and anti-angiogenesis effect of RPE in vivo. RESULTS: RPE exerted a higher inhibition effect on the proliferation of HUVECs than Lovo cells. The tube formation and cell migration were also significantly inhibited in the presence of RPE in a concentration-dependent manner though the significant inhibition effects were observed at the cytotoxic dose. RPE induced cell apoptosis and G0-G1 cell cycle arrest of HUVECs. In vivo, significant tumor growth inhibition was observed in human colon cancer xenografts established by Lovo cells, accompanying by a marked decrease in MVD. CONCLUSIONS: Our current study exhibited that RPE has a selective cytotoxity against HUVECs comparing to Lovo cells and also demonstrated significant anti-angiogenic effect in vivo.

Synergistic anti-proliferative effects of gambogic acid with docetaxel in gastrointestinal cancer cell lines.

BACKGROUND: Gambogic acid has a marked anti-tumor effect for gastric and colorectal cancers in vitro and in vivo. However, recent investigations on gambogic acid have focused mainly on mono-drug therapy, and its potential role in cancer therapy has not been comprehensively illustrated. This study aimed to assess the interaction between gambogic acid and docetaxel on human gastrointestinal cancer cells and to investigate the mechanism of gambogic acid plus docetaxel treatment-induced apoptotic cell death. METHODS: MTT assay was used to determine IC(50) values in BGC-823, MKN-28, LOVO and SW-116 cells after gambogic acid and docetaxel administration. Median effect analysis was applied for determination of synergism and antagonism. Synergistic interaction between gambogic acid and docetaxel was evaluated using the combination index (CI) method. Furthermore, cellular apoptosis was analyzed by Annexin-V and propidium iodide (PI) double staining. Additionally, mRNA expression of drug-associated genes, i.e., ß-tublin III and tau, and the apoptosis-related gene survivin, were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). RESULTS: Gambogic acid provided a synergistic effect on the cytotoxicity induced by docetaxel in all four cell lines. The combined application of gambogic acid and docetaxel enhanced apoptosis in gastrointestinal cancer cells. Moreover, gambogic acid markedly decreased the mRNA expression of docetaxel-related genes, including ß-tubulin III, tau and survivin, in BGC-823 cells. CONCLUSIONS: Gambogic acid plus docetaxel produced a synergistic anti-tumor effect in gastrointestinal cancer cells, suggesting that the drug combination may offer a novel treatment option for patients with gastric and colorectal cancers.

The herb medicine formula "chong lou fu fang" increases the cytotoxicity of chemotherapeutic agents and down-regulates the expression of chemotherapeutic agent resistance-related genes in human gastric cancer cells in vitro.

The herb medicine formula "Chong Lou Fu Fang" (CLFF) has efficacy in inhibiting the proliferation of human gastric cancer in vitro and in vivo. To explore the potentially useful combination of CLFF with chemotherapeutic agents commonly used in gastric cancer therapy, we assess the interaction between CLFF and these chemotherapeutic agents in both SGC-7901 cell lines and BGC-823 cell lines using a median effect analysis and apoptosis analysis, and we also investigate the influence of CLFF on chemotherapeutic agent-associated gene expression. The synergistic analysis indicated that CLFF had a synergistic effect on the cytotoxicity of 5-fluorouracil (5-FU) in a relative broad dose inhibition range (20-95% fraction affected in SGC-7901cell lines and 5-65% fraction affected in BGC-823 cell lines), while the synergistic interaction between CLFF and oxaliplatin or docetaxel only existed in a low dose inhibition range (≤50% fraction affected in both cell lines). Combination of CLFF and chemotherapeutic agents could also induce apoptosis in a synergistic manner. After 24 h, CLFF alone or CLFF combination with chemotherapeutic agents could significantly suppress the levels of expression of chemotherapeutic agent resistance related genes in gastric cancer cells. Our findings indicate that there are useful synergistic interactions between CLFF and chemotherapeutic agents in gastric cancer cells, and the possible mechanisms might be partially due to the down-regulation of chemotherapeutic agent resistance related genes and the synergistic apoptotic effect.

Proteomic analysis of anti-tumor effects by Rhizoma Paridis total saponin treatment in HepG2 cells.

Rhizoma Paridis total saponin (RPTS) had been identified as the major components responsible for the anti-tumor effects of the herb Rhizoma Paridis, which had been used in China for centuries to treat many diseases including tumor. To elucidate the anti-tumor mechanism of RPTS, a proteomic analysis was carried out with RPTS treatment in HepG2 cells. More than 50 proteins showed a significant change between control (0.01% DMSO) and RPTS (IC(50) approximately 10microg/ml) treated cells after 48h. Twelve proteins had been identified by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) using peptide fingerprinting from 15 protein spots (density difference >2 fold between the control and RPTS-treated group). Among them, six proteins were down-regulated (dUTPase, hnRNP K, GMP synthase, etc.) and six proteins were up-regulated (DNase gamma, Nucleoside diphosphate kinase A, Centrin-2, etc.) by RPTS treatment in HepG2 cells as determined by spot volume (p<0.05). Most of the identified proteins were associated with tumor initiation, promotion, and progression. These findings might offer valuable insights into the mechanism of anti-tumor effect affected by RPTS treatment in HepG2 cells.

In vitro anticancer activity of aqueous extracts and ethanol extracts of fifteen traditional Chinese medicines on human digestive tumor cell lines.

The purpose of this study was to investigate the anticancer activity of 15 traditional Chinese medicines which are usually used for tumor patients in China. The MTT (methylthiazolyldiphenyl-tetrazolium bromide) method was applied to compare the antitumoral activity of the aqueous crude extracts and the ethanol crude extracts of these drugs on six human digestive tumor cell lines: human liver carcinoma cell lines (HepG-2 and SMMC-7721), human gastric cancer cell line (BGC-823), human colon adenocarcinoma cell lines (LoVo and SW-116) and esophagus adenocarcinoma cell line (CaEs-17). Most ethanol extracts demonstrated a more powerful inhibitory effect than aqueous extracts. Their IC50 values were between 10 microg/mL and 500 microg/mL. Among these drugs, Paris polyphylla Smith showed a predominant inhibitory effect on all the cell lines with IC50 values ranging from 10 microg/mL to 30 microg/mL. The findings in this study suggested that traditional Chinese medicines, especially Paris polyphylla Smith, might have potential anticancer activity on digestive cancer and its mechanism needs further study.

[Inhibitory effect of oxaliplatin in combination with hyperthermia on angiogenesis].

OBJECTIVE: To study the effect of oxaliplatin in combination with hyperthermia on angiogenesis in vitro and in vivo. METHODS: MTT method was used to observe the influence of oxaliplatin on the proliferation of human umbilical vein endothelial cells (HUVEC) or human colon cancer cells (LOVO). The influence of oxaliplatin on HUVEC migration was evaluated by Transwell. Chick embryo chorioallantoic membrane (CAM) model was used to check whether the neovascularization of CAM could be suppressed in vivo. RESULTS: The survival rate of HUVEC was 80.1% - 42.5% within a range of 0.5 - 16 microg/ml and was negatively correlated with the concentration (correlation coefficient was - 0. 943, P = 0.005). The survival rate of LOVO cells within those doses was more than that of HUVEC. There was a synergistic antiangiogenic effect when a combination of oxaliplatin (0.5 microg/ml, 1 microg/ml and 16 microg/ml) with hyperthermia was used while additional effect was shown by the combinatioin of oxaliplatin (2 microg/ml, 4 microg/ml and 8 microg/ml) and hyperthermia in vitro. Oxaliplatin inhibited migration of HUVEC in vitro at low doses (0.25 - 2 microg/ml), and also suppressed angiogenesis of CAM in vivo at doses of 1 -4 microg/ml. CONCLUSION: The results of this experiment showed that low dose of oxaliplatin has anti-angiogenic effect in vitro, while in combination with hyperthermia has additional effect both in vivo and in vitro.