Objective:To construct a replication-deficient adenovirus carrying p16 gene and to investigate its anti-tumor activity on human gastric cancer xenografts in nude mice.Methods:p16 cDNA was amplified by PCR and was inserted into the plasmid pSuCMV,the latter was then used to recombine the replication-deficient adenovirus AdCMV-p16 in 293 cells.Human SGC-7901 gastric cancer xenograft models were established in nude mice and were divided into 3 groups:AdCMV-p16,Ad-LacZ,and control groups.Mice in AdCMV-p16 group received intratumoral injections of 2?108 pfu/100 l AdCMV-p16(injected every other day for 5 times).Mice in control group received the same volume of virus preserving solution.The tumor volumes were measured at predefined time points.The anti-tumor effect of AdCMV-p16 was observed by p16 immunochemical study and TUNEL detection of cell apoptosis.Results:The replication-deficient adenovirus expressing p16 gene evidently inhibited the growth of human gastric cancer xenografts in nude mice(P0.05),only with a inhibition rate of 4.26%.The pathological examination showed that apoptoses were the main changes in AdCMV-p16 group,and p16 gene was found in the cancer cells.Conclusion:The replication-deficient adenovirus harboring p16 gene can recover the expression of p16 in gastric cancer cells and subsequently inhibit the growth of human gastric cancer.

OBJECTIVE: To investigate the expression and significance of hypoxia inducible factor-2α (HIF-2α) and transcription factor OCT-4 in non-small cell lung cancer (NSCLC), and evaluate their roles in the prognosis of NSCLC. METHODS: Tissues from 51 cases of NSCLC were collected and immunohistochemistry (SP method) was used to detect the expression of HIF-2α and OCT-4 proteins. The correlation between the protein expression and the prognosis of NSCLC was analyzed. RESULTS: The positive rates of HIF-2α and OCT-4 expression in the NSCLC were 52.9% and 72.5%, respectively. There was significant relation between the expression of HIF-2α and OCT-4 (r=0.514,P<0.01). High expression of them revealed poor prognosis for NSCLC patients characterized with a bad overall survival(P<0.05). CONCLUSION There is a negative corelation between the expression of HIF-2α and OCT-4 and the prognosis of NSCLC. Combined examination of HIF-2α and OCT-4 expression might be an important biomarker for NSCLC prognosis.
Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Biomarkers, Tumor ; metabolism ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; metabolism ; Female ; Humans ; Lung Neoplasms ; diagnosis ; metabolism ; Male ; Octamer Transcription Factor-3 ; metabolism ; Prognosis

DNA barcoding is a rapidly developing frontier technology in the world and will be useful in promoting the quality control and standardization of traditional Chinese medicine. Until now, many studies concerning DNA barcoding have focused on leaf samples but rarely on Chinese herbal medicine. There are three issues involved in DNA barcoding for traditional Chinese medicinal materials: (1) the extraction methods for total DNA of the rhizomes of the medicinal materials; (2) intra-specific variation among samples from different places of origin; (3) accuracy and stability of this method. In this study, Gentianae Macrophyllae Radix was used to verify the stability and accuracy of DNA barcoding technology. Five regions (ITS2, psbA-trnH, matK, rbcL, and ITS) were tested for their ability to identify 86 samples of Gentianae Macrophyllae Radix and their adulterants. After improving the DNA extraction method, genomic DNA from all samples was successfully obtained. To evaluate each barcode's utility for species authentication, PCR amplification efficiency, genetic divergence, and species authentication were assessed. Among all tested regions only ITS2 locus showed 100% of PCR amplification and identification efficiencies. Based on the established method, we successfully identified two samples of Gentianae Macrophyllae Radix bought in pharmacy to the original species.

OBJECTIVE: To provide reference for standardizing clinical pharmacist training.METHODS: According to actual situation in our hospital, a new clinical pharmacist training teaching method [lecture-based learning (LBL) -case-based learning (CBL) combination method] was created by integrating the training resources of clinical teaching and research department and extending teaching content. The comprehensive skills of trainees after training were compared between 2015 (traditional one-to-one teaching model) and 2016 (LBL-CBL combination method). RESULTS & CONCLUSIONS: In 2016, trainees are significantly improved in the ability to analyze (3. 8 score vs. 2. 8 score) and deal with clinical problems (2. 9 score vs. 2. 5 score) compared with those in 2015. LBL-CBL combination method can satisfy the demands of clinical pharmacists training in our hospital. Not only basic theory and clinical knowledge of trainees are improved, but also clinical thinking and comprehensive skill of trainees are strengthened by using LBL-CBL combination method. LBL-CBL combination method can be used in clinical pharmacists training and its usual tests, and contributes to the improvement of clinical pharmacist training teaching.

Objective To investigate the associations of IFNL3/IFNL4 single-nucleotide polymorphisms(SNPs)with the efficacy of highly active antiretroviral therapy(HAART)in patients with HIV-1 infection.Methods Sixty-three adult patients with HIV-1 infection receiving HAART for at least 1 year in the First Affiliated Hospital, Zhejiang University School of Medicine were enrolled.HIV-1 RNA loads in plasma and HIV-1 DNA loads in peripheral blood mononuclear cells(PBMCs),and blood SNPs were detected by quantitative polymerase chain reaction(qPCR).Plasma inflammatory cytokines were examined by magnetic beads method,and the CD4 +T and CD8 +T lymphocyte counts in peripheral blood were measured by flow cytometry.According to response to HAART,the patients were classified as low HIV-1 RNA group(viral load <100 copies/mL)and high HIV-1 RNA group(viral load≥100 copies/mL);according to CD4+T lymphocyte counts,the patients were defined as low CD4+T cell group(<250 cells/μL), and high CD4+T cell group(≥250 cells/μL);according to HIV-1 DNA levels,the patients were divided into low(<100 copies/106cells)and high(≥100 copies/106cells)HIV-1 DNA groups.Results Three candidate SNPs rs368234815,rs8099917 and rs4803223 had significantly different distribution between low and high HIV-1 RNA groups(χ2=0.043,0.047 and 0.032,all P<0.05).The levels of interleukin(IL)-10 were declined in the low HIV-1 RNA group(U=4.00,P<0.05);the levels of IL-13 were decreased in the high HIV-1 RNA group and the high HIV-1 DNA group(U=0.00 and 2.00,both P<0.05);the levels of IL-21 were reduced in the high HIV-1 RNA group and in the low CD4 +T cell group(U=3.00 and 2.00, both P<0.05),the levels of IL-28A were decreased in the high HIV-1 RNA group,the high HIV-1 DNA group,and the low CD4 +T cell group(U=3.00, 0.50 and 3.00,P<0.05 or <0.01).In addition, rs368234815 was associated with IL-21 level(H=6.690,P<0.05),the IL-21 level in rs368234815 ΔG/ΔG [131.88(2.66,174.00)]was higher than that in TT/TT[6.79(2.81,26.48)](P<0.05);rs4803223 was correlated with IFN-γlevel(H=6.690, P<0.05),the IFN-γlevel in GG subtype[62.26(19.45, 96.49)]was higher than that in GA subtype[6.98(2.19, 99.14)](P<0.05).Conclusion The polymorphisms of IFNL3/IFNL4 rs368234815, rs8099917 and rs4803223 are associated with efficacy of HAART and immune-associated cytokines levels in patients with HIV-1 infection.

Objective:To construct the prognostic prediction model and scoring tool by using severe trauma patients’ physiological indicators on admission, and to verify the clinical application effect and provide a reference for the early evaluation of severe trauma patients.Methods:This study was a retrospective study which adopted cluster sampling. Patients who met the inclusion and exclusion criteria in the emergency department of the First Affiliated Hospital of Soochow University from September 2019 to November 2020 were included. Patients were randomly assigned into the modeling group and the validation group in a ratio of 7:3 based on their outcome in the emergency department. Logistic regression analysis was performed to construct a prediction model, which was simplified as a scoring tool. The model was verified by using validation group and two months’ prospective validation. The efficiency of the simplified scoring tool was compared with that of the revised trauma score (RTS) and the injury severity score (ISS).Results:Totally 863 patients were included in this study, including 604 patients in the modeling group and 259 patients in the validation group. The model included systolic blood, SpO 2 and AVPU score. The AUC for predicting the death of severe trauma patients was 0.938. The AUC of the prediction model was 0.933, the best cut-off point was 5, the sensitivity was 86.7%, the specificity was 94.2%; the AUC of the validation was 0.885, the sensitivity was 83.3%, the specificity was 93.7%; and the AUC of prospective validation was 0.919, the sensitivity was 100%, and the specificity was 76.7%. The AUC of the RTS and ISS were 0.800 and 0.833, respectively. The AUC of RTS was lower than that of the simplified scoring tool constructed in this research. Conclusions:The prediction model and simplified scoring tool are better than RTS in predicting the outcome of emergency severe trauma patients, which are convenient for emergency medical staff to evaluate the severity of trauma patients.

Objective:To explore the value of nomogram based on arterial spin labeling (ASL) MRI perfusion parameters and clinicopathological features in predicting the response to chemoradiotherapy (CRT) in advanced nasopharyngeal carcinoma (ANPC, stage Ⅲ and Ⅳ).Methods:From June 2018 to January 2021, 70 patients with ANPC confirmed by pathology were prospectively enrolled in Affiliated Hospital of Jiangnan University. Nasopharyngeal MRI plain scan, ASL and contrast-enhanced scan were performed before CRT, and routine MRI re-examination was performed within 1 week after the end of CRT. The pre-CRT perfusion parameter tumor blood flow (TBF) from ASL and clinicopathological features were recorded, and the maximum diameter (MD) of the tumor on T 1WI images was measured. The patients were divided into CRT effective group (48 cases) and ineffective group (22 cases) according to the response evaluation criteria in solid tumors. The independent sample t test was used to compare the differences of TBF, age and MD between effective group and ineffective group. The χ 2 test was used to compare the differences of gender, clinical stage and pathological type between the 2 groups. Using binary logistic regression analysis, clinicopathological model and TBF combined clinicopathological model were constructed, and the nomogram of combined model was constructed. The diagnostic efficacy of the models was obtained by receiver operating characteristic (ROC) curve analysis, and the area under the ROC curves (AUC) of the 3 models were compared by DeLong method. The calibration curve for the nomogram was generated, and the concordance index (C index) was acquired. Results:The TBF of the effective group and the ineffective group were (113±9) and (97±14) ml·100 g -1·min -1, with a statistical difference ( t=5.17, P<0.001). The MD value of the effective group was smaller than that of the ineffective group, with a statistical difference ( t=-2.24, P=0.028). There were statistical differences in clinical stage and pathological type between the 2 groups (χ 2 values were 12.21 and 12.95, respectively, both P<0.001). Three independent predictors, including TBF (OR=7.749), clinical stage (OR=0.129) and pathological type (OR=5.228), were included in logistic regression analysis. The AUC, sensitivity and specificity of TBF model in predicting the response to CRT were 0.843, 87.5% and 72.7%, of clinicopathological model were 0.822, 80.2% and 59.1%, of the nomogram model were 0.893, 81.2% and 90.9%. There was no statistical difference of AUC between the nomogram model and TBF model ( Z=1.23, P=0.215). However, the AUC of the nomogram model was greater than that of the clinicopathological model ( Z=2.47, P=0.031). The calibration curve showed that there was a good concordance index (C index=0.892) between the predicted value of nomogram and the actual clinical observation value. Conclusion:TBF, clinical stage and pathological type are independent predictors of the response to CRT in ANPC patients, and the nomogram based on these three factors has a good ability in predicting the response to CRT.

Objective:To construct a postoperative exercise management program for elderly patients with lung cancer based on social cognitive theory, and to provide guidance for improving the postoperative exercise ability of elderly patients with lung cancer.Methods:Systematically searched UpToDate, PubMed, Web of Science, Cochrane Library, Medlive, Wanfang, CNKI and other databases for relevant literature on postoperative rehabilitation exercise for elderly patients with lung cancer, with the search time from the establishment of the database to February 13, 2023. Guided by social cognitive theory, a preliminary draft of the postoperative exercise management program for elderly patients with lung cancer was prepared based on the literature research, and the final draft was formed after revising the program content through expert meeting method.Results:The questionnaire recovery rate of expert meeting method was 12/12; the expert authority coefficient was 0.892; the importance coefficient of variation of each item was 0-0.150; the Kendall coordination coefficient was 0.262 ( P<0.001); the final exercise management program included 5 first-level items, 18 second-level items and 42 third-level items. Conclusions:The process of constructing the postoperative exercise management program for elderly patients with lung cancer has scientificity and reliability, and the content has rationality and comprehensiveness, which can provide guidance for improving the postoperative exercise rehabilitation of elderly patients with lung cancer.

Objective:To find new biomarkers for the diagnosis and prognosis of sepsis through analyzing the differential expression protein in sepsis by proteomics and bioinformatics analysis and enzyme linked immunosorbent assay (ELISA).Methods:Patients with sepsis admitted to the emergency department of the Affiliated Hospital of Southwest Medical University from January to December 2019 were enrolled. And meanwhile, healthy volunteers who had normal physical examinations were included as the control group. Blood samples from two groups were collected. The samples were randomly selected for the protein concentration by data independent acquisition (DIA). Bioinformatics method was used in differentially expressed proteins by gene ontology (GO) pathway, enrichment analyses, groups meta-analysis and survival curves construction. ELISA method was used to verified marker screened. Then the data of transferrin receptor CD71 and the clinical data of procalcitonin (PCT), C-reactive protein (CRP) and blood lactic acid (Lac) were collected to construct receiver operator characteristic curve (ROC curve), and biomarker was screened for diagnostic and prognostic of sepsis.Results:The result of DIA showed that 71 differentially expressed proteins were screened out from sepsis group, 6 proteins were down-regulated and 65 proteins were up-regulated. Those differentially expressed proteins were enriched in the inflammatory response, response to stress, leukocyte migration in the GO pathway and enrichment analyses. The meta-analysis showed that the expression level of CD71 was higher in sepsis group than normal control group [standardized mean difference ( SMD) = -0.47, 95% confidence interval (95% CI) was -0.93 to 0.00, P < 0.01], the expression level of CD71 was higher in non-survivor group than survivor group ( SMD = -0.44, 95% CI was -0.70 to -0.18, P = 0.63). Survival curve showed that the expression of CD71 was inversely correlated to survival rates, the patients with a lower expression had higher survival rates ( P = 0.000 34); the ELISA showed that the level of CD71 was higher in sepsis group than normal control group (nmol/L: 156.83±84.71 vs. 87.99±47.89, P < 0.05), the level of CD71 was higher in non-survivor group than survivor group (nmol/L: 219.63±125.59 vs. 130.97±40.45, P < 0.05). The area under the ROC curve (AUC) of CD71 in diagnostic performance of sepsis was 0.790 (sensitivity was 65.1%, specificity was 90.0%), the AUC of CD71 in prognostic performance of sepsis was 0.744 (sensitivity was 57.1%, specificity was 94.1%); CD71 had a better prognostic performance than PCT (AUC = 0.547, sensitivity was 64.3%, specificity was 55.9%), CRP (AUC = 0.594, sensitivity was 64.3%, specificity was 61.8%), Lac (AUC = 0.540, sensitivity was 42.9%, specificity was 82.4%). Conclusion:CD71 had a great value of diagnostic and prognostic performance in sepsis, and it was expected to be a potential biomarker for sepsis.