[Identification of cellular morphology and flow cytometry on different tetraploid Isatis indigotica strains].

OBJECTIVE: To identify tetraploid Isatis indigotica strains through morphology and flow cytometry. METHODS: The tissue culture seedlings of tetraploid Isatis indigotica were root-tip squashed and chromosome counted before rooted climatized and transplanted in field. The plants in field were taken as experimental materials. Macroscopic observation was applied to identify by form and structure; Free-hand section was used to observe the length, width and density of stomas; And flow cytometry was applied to identify the ploidy. RESULTS: Compared with diploid plants, tetraploid plants had obvious changes in form and structure. The stomas from the tetraploid were notably longer, and the number of guard cells in chloroplasts was remarkably larger. The experiment materials were proved to be tetraploid by flow cytometry. CONCLUSION: The materials are tetraploid plants. Macroscopic observation, the length of stoma and the number of guard cells in chloroplasts can be taken as aided identification for ploidy of mutagenesis materials. Meanwhile, flow cytometry can be applied to identify the ploidy of Isatis indigotica.

[Effects of PEG and NaCl stress on the contents of indigotin and indirubin in Isatis indigotica tetraploids].

OBJECTIVE: To provide a reference evidence for choosing salt-resistent and drought-resistent varieties of Isatis indigotica Fort.. METHODS: The tissue culture seedlings of different Isatis indigotica tetraploids were used as plant materials. The diploid was used as CK, which were cultured for 30 days. 2 g/L NaCl and 15% PEG-6000 were used as stress treatment respectively for 12 hours, then poured out the solution, the materials were continuously cultured for 7 days. The contents of indirubin and indigotin were determined by HPLC. RESULTS: The contents of indigotin and indirubin of DB3, DB5 and DB12 with PEG treatment and DB2, DB3 and DB12 with NaCl treatment were increased. CONCLUSION: DB3 and DB12 can be further studied as reproduction materials.

[Initial RAPD analysis of Isatis indigotica somatic clones' resistance to root rot disease].

OBJECTIVE: To make initial identification on Isatis Indigotica Somatic Clones' resistant mutant. METHODS: The resistant plants were selected under the treatment of root-rot disease crude toxin. The DNA of the same clones and their parents were analysed by RAPD technology. RESULTS: We got initial resistance of the plants and selected 12 primers of which most were identical but the less were different. The genetic background of the somatic clones was identical but the individuals were mutated on the DNA level. CONCLUSION: It provides experimental foundation for further study on separating resistant mutant and resistant gene.

[Study on the extraction of total flavonoids from Mentha spicata by ultrasonic method].

The optimal conditions of total flavonoids extraction from Mentha spicata were identified by orthogonal test. The results showed that the ethanol concentration and the ultrasonic time had significant effect on the extraction of total flavonoids and the optimum conditions of extraction were A3B3C1D1, which were as follows: the ethanol concentration was 50%, the ultrasonic time was 40 min, the solid-liquid ratio was 1 g: 50 mL and the temperature of extraction was 60 degrees C, On the basis of the extracting method above, the content of total flavonoids in Mentha spicata was 0.0951 mg/mg. The linear equation was A = 11.51143C - 0.00029, r = 0.9999. This method is simple, quick and accurate with good prosperity.

Neurotensin promotes cholangiocarcinoma metastasis via the EGFR/AKT pathway.

Cholangiocarcinoma (CCA) is a fatal disease with increasing morbidity and poor prognosis due to poor response to conventional chemotherapy or radiotherapy. Neurotensin (NTS) has long been recognized as an important factor in the central nervous system and as an endocrine agent in the peripheral circulation via NTS receptor (NTSR) mediated actions. In recent years, NTS has been implicated in the carcinogenesis of numerous cancers; however, its role in cholangiocarcinoma remains obscure. Here, we observed the expression of NTS in cholangiocarcinoma vs. non-cancerous tissues and found that up-regulation of NTS facilitated cholangiocarcinoma cell metastasis and down-regulation of NTS inhibited their migration ability. Mechanistically, NTS drove cholangiocarcinoma cell metastasis via the EGFR/AKT pathway. Both the PI3-K inhibitor LY294002 or EGFR inhibitor Erlotinib stopped the discrepant metastatic capacity between NTS-depleted cholangiocarcinoma cells and control cells, further confirming that EGFR/AKT was required in NTS-promoted cholangiocarcinoma cell metastasis. More importantly, overexpression of NTS predicted poor prognosis of CCA patients. In summary, NTS could promote cholangiocarcinoma cells metastasis by amplifying EGFR/AKT signaling and may therefore be useful to predict patient prognosis.

Transcription factor WRKY70 displays important but no indispensable roles in jasmonate and salicylic acid signaling.

The transcription factor WRKY70 was previously reported to be a common component in salicylic acid (SA) and jasmonate (JA) mediated signal pathways in Arabidopsis. Here, we present that the inactivation of the WRKY70 gene in wrky70-1 mutant does not alter the responses of both JA and SA, and that wrky70 mutation is unable to restore the coi1 mutant in JA responses. However, overexpression of WRKY70 reduces JA responses such as expression of JA-induced genes and JA-inhibitory root growth, and activates expression of SA-inducible PR1. These data indicate that the WRKY70 is important but not indispensable for JA and SA signaling, and that other regulators may display the redundant role with WRKY70 in modulation of JA and SA responses in Arabidopsis. Furthermore, we showed that JA inhibits expression of WRKY70 and PR1 by both COI1-dependent and COI1-independent pathways.

[Determination of trace lead in traditional Chinese herbal medicine Astragalus by microwave digestion-CTAB enhancing-continual flow ingection hydride generation-ICP-AES].

A new method using microwave digestion technique was developed for the determination of lead in Astragalus by CTAB enhancing-continual flow hydride generation-inductively coupled plasma atomic emission spectrometry (HG-ICP-AES). The experimental conditions of microwave digestion and hydride generation were optimized. This method shows a linear range of 0.23-800 microg x L(-1) and the correlation coefficient is 0.999 9. It is satisfactory to apply the microwave digestion procedure to the determination of Pb under the optimized conditions. The detection limit of the method is 0.23 microg x L(-1) and the RSD is 1.02%. The recovery obtained is 98.8%-100.1%. The results show that this method is rapid and simple with low environmental contamination and complete digestion of samples.

Glasgow Prognostic Score predicts prognosis of intrahepatic cholangiocarcinoma.

High Glasgow Prognostic Score (GPS) has been associated with poor prognosis in patients with lung, ovarian, colorectal and renal cancer, as well as hepatocellular carcinoma. The aim of this study was to investigate the prognostic value of GPS in patients with intrahepatic cholangiocarcinoma (ICC) undergoing partial hepatectomy. A total of 72 patients with pathologically confirmed ICC were classified according to their GPS scores assigned based on the preoperative levels of C-reactive protein (CRP) and albumin. Their clinicopathological data were retrospectively assessed using univariate and multivariate analysis to determine their association with overall survival and recurrence. High GPS scores in ICC patients were associated with preoperative levels of CRP (P<0.001) and albumin (P<0.001), frequency of ascites accumulation (P=0.035), lymph node metastasis (P=0.002) and tumour size (P=0.005). On univariate analysis, preoperative levels of CRP (P<0.001), albumin (P=0.016) and carbohydrate antigen 19-9 (P=0.038), hepatitis B virus (HBV) positivity (P=0.009), occurrence of lymph node metastasis (P=0.001), Child-Pugh class B (P=0.013) and high tumour-node-metastasis (TNM) stage (P=0.002) were found to be associated with the 1- and 3-year overall survival. Multivariate analysis suggested that GPS score (HR=2.037, 95% CI: 1.092-3.799, P=0.025), TNM classification (HR=2.000, 95% CI: 1.188-3.367, P=0.009) and HBV positivity (HR=0.559 95% CI: 0.328-0.953, P=0.032) were independently associated with patient survival. High GPS scores also predicted ICC recurrence. In conclusion, our results demonstrated that GPS may serve as an independent marker of prognosis in patients with ICC following partial hepatectomy.

[Study on induction of polyploidy in Salvia bowleyana by colchicine treatment].

OBJECTIVE: To explore the technique of induction of polyploidy in Salvia bowleyana by colchicine treatment. METHOD: The three kinds of explant of bud, leaf and calli were induced by colchicine treatment. RESULT: The induction effects were better when the calli was treated by colchicines (15 mg x L(-1)) and the leaf was pre-cultured for one week. The doubling rate was 33.33%, while the majority were wholy doubled plants, and the leaves were thicker and broader, the color was darker, the root was thicker and the stoma size was obviously bigger than the diploid plants. The number of chromosome were 8 to 64. Isoenzyme analysis showed that the enzyme activities between the polyploid and the diploid plants were quite different. CONCLUSION: Induction of polyploidy by colchicine treatment is efficacious. The part of the doubled plants were identified as homologmous tetraploids.

A comparison of the prognostic value of preoperative inflammation-based scores and TNM stage in patients with gastric cancer.

BACKGROUND: People's Republic of China is one of the countries with the highest incidence of gastric cancer, accounting for 45% of all new gastric cancer cases in the world. Therefore, strong prognostic markers are critical for the diagnosis and survival of Chinese patients suffering from gastric cancer. Recent studies have begun to unravel the mechanisms linking the host inflammatory response to tumor growth, invasion and metastasis in gastric cancers. Based on this relationship between inflammation and cancer progression, several inflammation-based scores have been demonstrated to have prognostic value in many types of malignant solid tumors. OBJECTIVE: To compare the prognostic value of inflammation-based prognostic scores and tumor node metastasis (TNM) stage in patients undergoing gastric cancer resection. METHODS: The inflammation-based prognostic scores were calculated for 207 patients with gastric cancer who underwent surgery. Glasgow prognostic score (GPS), neutrophil lymphocyte ratio (NLR), platelet lymphocyte ratio (PLR), prognostic nutritional index (PNI), and prognostic index (PI) were analyzed. Linear trend chi-square test, likelihood ratio chi-square test, and receiver operating characteristic were performed to compare the prognostic value of the selected scores and TNM stage. RESULTS: In univariate analysis, preoperative serum C-reactive protein (P<0.001), serum albumin (P<0.001), GPS (P<0.001), PLR (P=0.002), NLR (P<0.001), PI (P<0.001), PNI (P<0.001), and TNM stage (P<0.001) were significantly associated with both overall survival and disease-free survival of patients with gastric cancer. In multivariate analysis, GPS (P=0.024), NLR (P=0.012), PI (P=0.001), TNM stage (P<0.001), and degree of differentiation (P=0.002) were independent predictors of gastric cancer survival. GPS and TNM stage had a comparable prognostic value and higher linear trend chi-square value, likelihood ratio chi-square value, and larger area under the receiver operating characteristic curve as compared to other inflammation-based prognostic scores. CONCLUSION: The present study indicates that preoperative GPS and TNM stage are robust predictors of gastric cancer survival as compared to NLR, PLR, PI, and PNI in patients undergoing tumor resection.

The Glasgow Prognostic Score is an independent prognostic predictor of hepatocellular carcinoma following radical resection.

BACKGROUND: Some prognostic evaluation systems were developed to postoperatively predict the outcome of hepatocellular carcinoma (HCC) patients, mainly based on the cancer itself and the underlying liver diseases. However, none of these prognostic evaluation systems have so far been universally accepted. A simple and feasible scoring system is still lacking for the prediction of prognosis of HCC patients following resection. We aimed to uncover the correlation between the preoperative Glasgow Prognostic Score (GPS) and the clinical outcome of HCC patients after radical resection. METHODS: The patients were separated into 3 subgroups on the basis of their GPSs. The prognostic significance of the GPS in the patient cohort was evaluated by survival analysis. RESULTS: On univariate analysis, the levels of C-reactive protein and albumin, the Child-Pugh class, vascular invasion, tumor number, tumor size, the tumor/node/metastasis (TNM) stage, and the GPS were associated with overall survival and time to recurrence of HCC patients after radical resection. On multivariate analysis, the tumor size, albumin level, and GPS were independently associated with the outcome of HCC postoperatively. CONCLUSION: The GPS is an independent biomarker for prognostic prediction of HCC following radical resection.