AIM:To study the blocking effect of shRNA on the expression of PSMA gene in LNCaP cell line by using shRNA eukaryotic expression vector.METHODS:Three pairs of DNA templates coding shRNA,synthesized against PSMA and cloned into the vector pSilencer 2.1-U6-neo,which was named pSilencer 2.1-U6-neo-shRNA,were identified by restriction endonuclease digestion analysis and DNA sequencing.LNCaP cells were then transfected with these three pSilencer 2.1-U6-neo-shRNAs and the negative control pSilencer 2.1-U6-neo-NC.After G418 selection,the cells were selected and the interfering effect was detected by RT-PCR and Western blotting.The biological behaviours of the transfected LNCaP cells were also tested.RESULTS:Restriction endonuclease digestion analysis and DNA sequencing results all showed that the 3 target segments were cloned into pSilencer 2.1-U6-neo vector respectively.After transfected into LNCaP cells,the inhibitory ratio of PSMA mRNA was 33.15%,9.26% and 41.97% respectively,and that of PSMA protein was 26.26%,6.47%,40.69% respectively.The p-shRNA3 was chosen to test the cell growth and its invasive power in vitro.The results showed that after interfering,the invasiveness of LNCaP cells were enhanced.CONCLUSION:The vector-based shRNA on PSMA gene effectively knocks down the PSMA gene expression.The successful construction of PSMA shRNA makes it possible for further study of the interaction between PSMA and prostate cancer.

Objective To understand the impairment and compensation mechanism of brain function in pa-tients with non-fluent aphasia after ischemic stroke. The fractional amplitude of low frequency fluctuation (fALFF) method was used to analyze the functional magnetic resonance imaging (fMRI) data in the resting state between the aphasia patients and the normal controls. Methods The scans of the resting state of fMRI were performed in 17 aphasia patients and 19 age-, education-, and sex-matched healthy control subjects. The scan sequence was single-shot echo planar image,DPARSF software was used to analyze fALFF data of the aphasia patients and the healthy controls. Results Compared to the control group, the value in right superior temporal gurus, inferior parietal lob-ule, frontal lobe cortex, and postcentral gurus were significantly increased in the aphasia group. The fALFF in bilat-eral cerebellum and right thalamus were also decreased in the aphasia group. Conclusions The fALFF values in some brain region in the aphasia group were abnormal in the resting state , indicating a few pathological change of brain function in patients with non-fluent aphasia after ischemic stroke.

Objective To study the clinical outcome of patients with fns-like tyrosine kinase-3internal tandem duplication (FLT3-ITD) positive acute myeloid leukemia (AML) treated with allogeneic hematopoietic stem cell transplantation (allo-HSCT) and to explore the potential prognostic factors to patients' survival including transplant types or disease status.Methods A total of 314 AML patients in our center from October 2006 to October 2012 were retrospectively analyzed,among whom,54 patients were defined with FLT3-ITD positive.Survival rates and treatment-related mortality were further analyzed.Results For all 54 FLT3-ITD positive patients,the 3-year overall survival (3-OS) rate was 56％ and 3-year leukemia-free survival (3-LFS) rate was 47％.The outcome of haplo-identical HSCT was similar as that of sibling donors(3-OS rate:60％ vs 54％ ; 3-LFS rate:54％ vs 45％,respectively).There were 47 patients who received transplantation in first complete remission(CR1).The other 7 patients were of disease relapse or in CR2 before transplantation.Not surprisingly,patients in CR1 had better prognosis than those in nonCR1.Conclusions Allo-HSCT is an effective treatment for AML patients with FLT3-ITD positive mutation.The survival outcome of haplo-identical HSCT was comparable with that of sibling donors.Relapse of AML was the dominant factor related to the mortality of FLT3-ITD positive AML patients after allo-HSCT.

Objective To investigate the regularity and predictors of weight loss during concurrent chemo-radiotherapy for nasopharyngeal carcinoma patients. Methods A retrospective study was carried out on 82 patients with nasopharyngeal carcinoma ,with average age of 45 years ,who received concurrent chemoradiotherapy at the Affiliated Cancer Hospital of Guangzhou Medical University. Paired t-test was used to analyze the weight loss pattern during chemoradiotherapy. The Spearman′s rank correlation analysis was used to investigate the correlation between high weight loss(HWL)and the clinical factors. Results The paired t-test showed that the weight of the patients decreased continuously every week during chemoradiotherapy(P < 0.05),and the value dropped most significantly during the 4th week. Among these 82 patients,32 were determined as HWL(weight loss > 10%). Spearman′s rank correlation analysis showed that HWL was associated with older age,female,higher T and TNM stage,3D-CRT technology,the higher dose of radiotherapy in nasopharynx and in neck lymph node,and decreases of leukopenia and hemoglobin over degree Ⅱ(P<0.05). Conlousions Weight may decrease continuously every week during chemoradiotherapy for nasopharyngeal carcinoma patients. Age,gender,T stage,TNM stage,radio-therapy dose,and leukopenia and hemoglobin decreases may be predictors for weight loss degree during concurrent chemoradiotherapy for nasopharyngeal carcinoma patients.

This study was designed to prove simultaneously blocking both EGFR and COX-2-medi-ated pathways may be an efficient means of inhibiting cancer cell growth in NPC. Method: A combination of tarceva (EGFR-selectivetyrosine kinase inhibitors) with celecoxib(Cox-2 inhibitor) was studied on its effects on cell growth, cell cycle progression, apoptosis and protein expression in CNE-2 cell lines by cell growth assay, flow cy-tometric analysis assay and Western blotting. Result: ①The inhibition rate of cell growth was higher in the group treated with a combination of two agents than that the sum of rates of the two groups treated with only one agent (P<0.05). ②The combination of tarceva with celecoxib significantly induced G_1 arrest(P<0.05) ,but did not in-crease apoptosis rate(P>0.05). ③The group of combination showed less expressions of p-EGFR and COX-2 than any other group. Conclusion Simultaneously blocking EGFR and COX-2 mediated pathways would significantly in-hibit the growth of CNE-2 cell line, increase G_1 arrest and reduce the expression levels of p-EGFR and COX-2.

Objective To assess under different vessel diameter ,the effect of the aspiration thrombectomy catheter in improving the myocardial reperfusion and clinical prognosis in patients with acute myocardial infarction (AMI)who were undergone primary percutaneous coronary intervention(PCI) .Methods 205 patients with AMI immediate implant stents after thrombus suction ,the TIMI flow grade(myocardial infarction thrombolysis treatment test flow classification ) ,postoperative ecg evolution ,incidence of no-reflow MACE in 30 days and MACE in 6 months were compared between conventional thrombus suction group and suction again group(blood vessels of <3 .0 mm and ≥3 .0 mm) .Results The level 3 blood flow rate ,MACE in 6 months in suction again group with blood vessels of ≥3 .0 mm had improved significantly ,but had no beneficial effects in blood vessels of ≥3 .0 mm .Conclusion In AMI patients treated with primary PCI ,application of aspiration thrombectomy catheter with blood vessels of ≥3 .0 mm may im-prove the flow condition before infarction related blood vessels ,reduce MACE .

AIM: To find out the gene structure and diversity of protate specific membrane antigen(PSMA) alternative spliced variants, and probe into the pathogenesis of prostate cancer.METHODS: 5'-RACE and 3'-RACE methods were used to amplify the 5' and 3'end of alternative spliced variant and then those viariants were sequenced for analyzing the gene stucture and diversity of PSMA alternative spliced variants of prostate cancer tissues.RESULTS: Four new alternative spliced variants of PSMA were discovered from prostate cancer tissues.Compared with reported PSMA alternative spliced variants,different insertions and deletions existed in different sites of those new variants.CONCLUSION: The discovery of the new variants confirms the diversity of PSMA spliced variants and provides the clues for seeking the target of diagnosis and therapy of prostate cancer.

Objective To explore the DNA damage in peripheral blood lymphocytes of rats exposed to sodium arsenite. Methods Thirty-two Wistar rats, weighing 180 - 200 g, equal male and female, were randomly divided into 4 groups, 8 in each group. Sodium arsenite 0(control) ,0.05,0.15,0.45 mg/L were given through drinking water for 30 days. Body weight and drinking water consumption were measured every day. Blood were collected and DNA damage in peripheral blood lymphocytes was examined by single cell gel electrophoresis.Results The increase of body mass[( 121.00 ± 38.57), ( 120.62 ± 42.80), ( 125.38 ± 48.68)g]and water intake [(36.9 ± 6.2), (37.9 ± 5.8), (39.3 ± 4.2)ml/d]in 0.05,0.15,0.45 mg/L sodium arsenite groups were compared with the control group[( 119.25 ± 47.27)g, (38.4 ± 5.1 )ml/d], and the difference were not significant (F = 0.040,0.828, all P ＞ 0.05). The tail ratios[46.25%(185/400) ,57.00%(228/400),64.00%(256/400)], tail lengths [(32.89 ± 17.18), (58.74 ± 36.28), (77.55 ± 35.73 ) μm]and tail moments [(6.29 ± 3.74), ( 11.20 ± 9.64),(17.30 ± 12.60)μm]in 0.05,0.15,0.45 mg/L sodium arsenite groups were significantly higher than those of the control group[39.25%(157/400), (18.73 ± 15.83),(2.61 ± 1.05)μm, all P ＜ 0.01], and the tail ratios,tail lengths and tail moments in lymphocytes increased with increased doses of arsenic concentration. Conclusions Low doses of arsenic exposure can induce DNA damage in peripheral blood lymphocytes of rats.

Objective: To learn the health literacy status, trend and associated factors of Ningbo residents from 2015 to 2019, so as to provide basis for developing health policies and interventions. Methods: The monitoring data of health literacy of Ningbo residents from 2015 to 2019 was collected. The health literacy level was calculated and standardized by the population data of the sixth national census of Ningbo in 2010; five-year absolute growth and growth rate were used to reflect the changing trend. Multivariate logistic regression model was employed to analyze the influencing factors for the health literacy level. Results: The health literacy levels from 2015 to 2019 were 15.44%, 21.73%, 22.41%, 27.60% and 30.03%, with an annual increase trend ( P<0.05 ). The five-year absolute growth and growth rate were 14.59% and 94.49%. The Results of multivariate logistic regression analysis indicated that the year ( OR=1.158, 95%CI: 1.132-1.184 ), age ( 25-<35岁 years, OR=1.235, 95%CI: 1.039-1.468; 35-<45岁 years, OR=1.416, 95%CI: 1.193-1.681; 45-<55岁 years, OR=1.221, 95%CI: 1.024-1.455 ) , education level ( primary school, OR=1.790, 95%CI: 1.461-2.195; junior high school, OR=2.574, 95%CI: 2.102-3.154; high school/vocational high school/technical secondary school, OR=4.863, 95%CI: 3.943-5.998; college or above, OR=8.829, 95%CI: 7.109-10.965 ) , urban areas ( OR=0.934, 95%CI: 0.874-0.998 ) and occupation ( farmers, OR=0.692, 95%CI: 0.608-0.787; workers, OR=0.746, 95%CI: 0.664-0.837; enterprise staff, OR=0.822, 95%CI: 0.745-0.906; others, OR=1.106, 95%CI: 1.009-1.211 ) were the influencing factors for health literacy level. Conclusion The health literacy level of Ningbo residents shows an upward trend from 2015 to 2019, which are associated with age, education level, area and occupation.

Objective To observe the protection and distribution of bone marrow mesenchymal stem cells (MSCs) by distinct intravenous infusion time on renal ischemia reperfusion injury (IRI) in rats.Methods We used unilateral nephrectomy and contralateral vascular occlusion method to establish renal IRI model in rats.The experimental groups which received 2 × 106 MSCs infusion through the tail vein,were subsequently divided into 3 subgroups:2 h pre-reperfusion (PreOp,n =16),immediately after reperfusion (Op,n =16),6 h post-reperfusion (PostOp,n - 16).The control groups included sham operation group (n =16) and ischemia group (n =16).Chemotaxis of DAPI-labeled MSCs was detected 6 h after administration in the IR kidney.Renal function was detected at 6,24,and 48 h respectively after operation. Forty eight h after operation,the renal tissues were harvested to observe the pathological changes by HE staining and the tubular epithelial cell apoptosis via TUNEL assay.Results MSCs were found in the experimental groups after IR in the kidney,most in PostOp group.Twenty-four and 48 h after reperfusion,there was no significant difference in Cr and BUN between the experimental groups and sham operation group (P＞0.05),but the levels of Cr and BUN in the experimental groups were significantly lower than in the IR group (P＜ 0.05). As compared with IR group,the renal pathological injury was alleviated,the number of apoptotic cells was decreased in the experimental group,most significantly in PostOp group (P＜0.05).Conclusion MSCs can reduce the inflammatory response and inhibit renal tubular cell apoptosis in rat renal IRI.Post-reperfusion administration of MSCs leads to the best chemotaxis efficiency and protection.