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Objective:To screen the endoplasmic reticulum stress (ERS) signature-related differentially expressed genes (DEG) in gastric cancer and to construct a prognostic risk model based on a bioinformatics.Methods:Transcriptome sequencing data (RNA-seq) of 375 gastric cancer and 32 paracancerous tissue samples downloaded from The Cancer Genome Atlas (TCGA) database and the corresponding clinical information were obtained as training set samples; data of 387 gastric cancer patients (GSE84437) from Gene Expression Omnibus (GEO) database were downloaded as validation set samples. All data were obtained on December 25, 2021. A total of 785 ERS signature-related genes (ERS-RG) were obtained from the GeneCards database. DEG between gastric cancer tissues and paracancerous tissues in the TCGA database was analyzed. The identified gastric cancer DEG were intersected with ERS-RG from the GeneCards database to obtain gastric cancer ERS signature-related DEG, which were analyzed for gene ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. Univariate Cox proportional risk model was used to screen ERS signature-related DEG with prognostic value in gastric cancer, and LASSO regression analysis was performed to construct a polygenic prognostic risk model, and to calculate the prognostic risk score. The patients in training set and validation set were divided into high-risk group and low-risk group according to the median of the prognostic risk score (2.369); Kaplan-Meier survival analysis was used to compare the overall survival (OS) and to draw time-dependent receiver operating characteristic (ROC) curves of patients in the two groups; nomogram was drawn based on the prognostic independent influencing factors of gastric cancer. The characteristic immune cell infiltration abundance between the two groups was analyzed by using the inverse convolution-based CIBERSORT algorithm. Cytolytic activity scores were calculated by using the geometric mean of granzyme A and perforin 1 expression. According to the median prognostic risk score (2.369) and median tumor mutation burden (TMB) (3.000), all patients with gastric cancer were divided into high risk score-high TMB group, high risk score-low TMB group, low risk score-high TMB group and low risk score-low TMB group to compare the OS of patients in each group.Results:A total of 444 ERS signature-related DEG in gastric cancer including 168 down-regulated genes and 276 up-regulated genes were obtained, which were mainly enriched in biological processes such as protein processing in the endoplasmic reticulum, extracellular matrix (ECM) receptor interactions and unfolded protein responses (all P < 0.05). Univariate Cox regression analysis showed that 12 prognostic-related ERS signature-related DEG in gastric cancer were screened out. LASSO regression analysis was performed to obtain a prognostic risk score = 0.052×NOS3+0.137×PON1+0.067×CXCR4+0.131×MATN3+0.116×ANXA5+0.090×SERPINE1. The results of Kaplan-Meier analysis showed that the OS of the low-risk group in both the training and validation sets was better than that of the high-risk group (all P < 0.01). The results of the time-dependent ROC curve analysis showed that the AUC for the 3-year, 5-year, 8-year OS rates was 0.695, 0.786, 0.698, respectively in the training set, while the AUC for the 3-year 5-year, 8-year OS rates was 0.580, 0.625, 0.627, respectively in the validation set. Multivariate Cox regression analysis showed that prognostic risk score ( HR = 3.598, 95% CI 2.290-5.655, P < 0.001) and tumor stage ( HR = 1.344, 95% CI 1.057-1.709, P < 0.05) were independent factors influencing the prognosis of gastric cancer. Among 375 gastric cancer patients in the TCGA database, the expression levels of ATF6, HSPA5, XBP1 and ATF4 in the high-risk group were higher than those in the low-risk group (all P < 0.05); CIBERSORT results showed that the abundance of activated CD4 memory T cells in the high-risk group was lower than that in the low-risk group, and the abundance of both M0 and M2 macrophages in the high-risk group was higher than that in the low-risk group (all P < 0.05). The expression levels of common immune checkpoints (CD274, CTLA4, TNFRSF9, TIGIT, PDCD1, LAG3) in the high-risk group were all higher than those in the low-risk group (all P < 0.05). Cytolytic activity score in the high-risk group was higher than that in the low-risk group ( P < 0.05). The prognostic risk score was negatively correlated with TMB ( r = -0.20, P < 0.001). Patients in the low-risk score-high TMB group had the best OS and those in the high-risk score-low TMB group had the worst OS (both P < 0.001). Conclusions:The prognostic risk score model is established based on 6 ERS signature-related DEG in gastric cancer and its prognostic risk score may be effective as an independent prognostic factor to predict the prognosis of gastric cancer patients.
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OBJECTIVE To establisha simultaneous determination method of three main alkaloids in Plumula nelumbinis by high performance liquid chromatography(HPLC), including liensinine, isoliensinine and neferine, and optinize the ultrasonic-assisted extraction process of total alkaloids from Plumula nelumbinis by using response surface methodology based on Box-Behnken design. METHODS The alkaloids were separated on a RP-HPLC C18 monolithic column(4.6 mm×50 mm, Merck, Darmstadt, German) with a mobile phase consisting of methanol-water-triethylamine-acetic acid(70∶30∶0.2∶0.05), and the method was validated for linearity, sensitivity and extraction recovery. Based on the single-factor analysis, a mathematical model was constructed to analyze the effects of each factor of the ultrasonic-assisted extraction and their interactions on alkaloids yields of Plumula nelumbinis. The three independent variables were volume fraction of ethanol(A), solid-liquid ratio(B) and ultrasound extraction time(C), respectively. RESULTS The HPLC method used to separate three alkaloids within 5 min showed an excellent linear correlation (r2>0.999) in the range of 3.4-340 μg·mL-1(liensinine), 3.47-347 μg·mL-1(isoliensinine) and 3.47-347 μg·mL-1 (neferine). The limits of detection(LODs) of three alkaloids were 0.17, 0.69 and 0.69 μg·mL-1, and the limits of quantification were 0.34, 1.73 and 1.73 μg·mL-1, respectively. The intra-day and inter-day variations of RSD were less than 5%, and the extraction recovery rang from 97.06% to 104.12% with RSD ranging from 0.08% to 0.87%(n=6). The optimum extraction conditions were: solid-liquid ratio, 1∶30.3; volume fraction of ethanol, 69.8%; ultrasound extraction time, 29.0 min. The yield under the optimum conditions was found to be(14.82±0.42) mg·g-1, which was agreed closely with the predicted value of 14.70 mg·g-1. CONCLUSION The HPLC method used to determine the three main alkaloids in Plumula nelumbinis shows ideal characteristics of quickness, accuracy, high sensitivity and good repeatability.The optimum ultrasonic extraction technology has expressed excellent extraction ability of alkaloids in Plumula nelumbinis which indicates the RSM result is reasonable and effective.
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Objective To investigate the safety of cleaning and disinfection of duodenoscope used in endoscopic retrograde cholangiopancreatography (ERCP).Methods The duodenoscopes,which were used in Endoscopy Center of Digestive Disease Hospital,Heilongjiang Provincial Hospital from May 4 to May 31 2016,were cleaned manually and disinfected with sterilization powder.The samples from elevator and elevator channel were collected for bacterial culture to evaluate the safety of clinical application of duodenoscope.Results Among 60 high level disinfections,one was excluded because disinfectant concentration wasn't effective.Among 59 elevator samples no positive cultures were detected,while among 59 channel samples,4(6.8％) cultures were positive and all from the same duodenoscope,including 3 cases of Klebsiella pneumonia and 1 case of Pseudomonas aeruginosa.Conclusion Sterilization powder is effective for disinfection of duodenoscope used for ERCP.Residual organic material can't be excluded from elevator sample even if it is negative for culture.Duodenoscope with high level disinfection still has the risk of transmitted infection,and should be monitored regularly by culture and reprocessed in time.
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Docetaxel (DXT)is a member of the taxane drug class,which is used to treat breast cancer,lung cancer,gastric cancer,prostate cancer and so on.Docetaxel can be used as a single agent or in combination with other chemotherapeutic drugs.Many side effects of docetaxel have been reported in recent years.New dosage forms are developed to reduce toxicity and increase efficacy.An overview of these novel formulations of docetaxel and clinical progress will be discussed by consulting 43 literatures.