2.The roles of RNA-editing enzyme ADAR1 in EV71 infection and virus mutation
Qingqing LIU ; Zhangmei CHANG ; Jinjin BAI ; Yan WANG ; Jianer LONG
Fudan University Journal of Medical Sciences 2017;44(3):253-260
Objective To identify the role of RNA-editing enzyme ADAR1 (adenosine deaminase acting on RNA) in EV71 infection and virus mutation.Methods RNAi technology was applied to establish ADAR1 knock-down stable cell lines.Then the cells were served to evaluate the role of ADAR1 in EV71 infection by MTT assay for detecting virus-induced cell viability,virus plaque assay for quantification of the virus titer and the cellular susceptibility to the virus,and Western blot for virus protein expressions.ADAR1-mediated RNA editing can result in the genetic A-G and T-C mutations.To further determine whether the effects of ADAR1 on EV71 infection were correlated with ADAR1-mediated EV71 RNA editing and therefore increased the viral mutations during the infection,the characteristics of EV71 mutation were analyzed based on the different full-length viral genomes from epidemic regions.The viral genome was also sequenced from the infected ADAR1 knock-down cells.Results After ADAR1 knock-down,the cell viability decreased quickly after the virus infection,and formed much more and larger sizes of plaques than the control cells.The virus capsid protein VP1 expressions and virus titer in the cells culture media were both increased in ADAR1 knockdown cells.Statistic analysis showed that A-G and T-C mutations were the major mutations of EV71,which were believed to be the hot sites for RNA-editing.However,the results of viral RNA genomic sequencing data indicated that ADAR1 did not edit EV71 genome directly.Conclusions ADAR1 was a restriction factor for controlling EV71.However,ADAR1 does not directly edit EV71 genome.
3. Optimization for vacuum belt drying process of Shiwei Penan Granule by using response surface methodology
Chinese Traditional and Herbal Drugs 2015;46(13):1914-1919
Objective: To optimize the vacuum belt drying process of Shiwei Penan Granules. Methods: On the base of single factor experiments, feeding rate, heating temperature, and drying time were selected as the main factors, with composite score of moisture content of dry paste, rates of polydatin and paeoniflorin transfer as the response values, response surface methodology was used to optimize the vacuum belt drying process. Results: Optimal parameters were as follows: The feeding rate was 1.6 kg/h, heating temperatures was 105℃, drying time was 104 min; Under these conditions, moisture content of dry extract was 2.80% on average, the transfer rate of polydatin was 92.55%, and the transfer rate of paeoniflorin was 94.77% on average, the composite score was 95.35 on average, RSD=0.42%. Conclusion: The condition optimized by using response surface methodology is stable and reasonable, which could guide industrial production and keep high transfer rate of effective component.
4.Serum CEA and CYFRA21-1 correlates with EGFR mutation and EGFR-TKI effectiveness in untreated NSCLC patients
Yujing SUN ; Chang LIU ; Yan LONG ; Yuanyuan SUN ; Chunhong FAN ; Hui WANG ; Xiaotao ZHAO
Chinese Journal of Laboratory Medicine 2015;(6):407-411
Objective To explore the predictive value of serum CEA and cytokeratin-19 fragments (CYFRA21-1)prior treatment for the epidermal growth factor receptor (EGFR) mutation and efficacy of tyrosine kinase inhibitors ( TKI ) in patients with non-small cell lung cancer.Methods The study was a clinical research.Totally 101 matched tissue and plasma samples were collected from Peking University People′s Hospital from 2012 to 2013.All clinical specimens were analyzed for EGFR mutations in exons of 18, 19, 20 and 21 by ADx-ARMS and direct sequencing, and the serum levels of CEA and CYFRA21-1 were analyzed by ECLI.The correlation between EGFR mutant status and efficacy of EGFR-TKI and clinicopathological parameters were analyzed by χ2 test, Log-rank text and Cox proportional hazards regression model.Results The mutation rate was 60.4%(61/101) by ADx-ARMS and 33.7%(34/101) by direct sequencing.Mutations were more frequently observed in the higher serum CEA level patients(≥5μg/L,78.8%).However, the rates of EGFR mutations of different CEA levels were similar.Among the patients receiving TKI therapy, the efficacy of EGFR-TKI was closely related to serum CYFRA21-1 level prior treatment and EGFR mutation (χ2 =8.903, P =0.003; χ2 =28.590, P <0.001 ).And serum CYFRA21-1 level prior treatment and EGFR mutation were independent factors for EGFR-TKI treatment affecting PFS (RR=0.298, P<0.001;RR=0.086, P<0.001).Conclusion The mutation rate of EGFR was significantly related with the expression level of CEA prior treatment, and serum CEA and CYFRA21-1 levels prior treatment could be potential predictors of EGFR-TKI efficacy.
5.The survey of pathology and survival rate for porcine models of penetrating abdominal firearm wound in normal temperature environment and dry heat environment
Jiangwei LIU ; Zexin LI ; Yongjiu ZHANG ; Xiaoping LONG ; Xiaoyun ZHAO ; Decong CHANG
Chinese Journal of Current Advances in General Surgery 1999;0(02):-
0.05) .The total survival rate of N group and H I group was significantly higher than H II group(P0.05) .The abdominal stinking leak was more severe and death rate was higher in N group and H I group than H II group.Conclusion:Dry heat environment in the desert could obviously impair the total survival rate of pigs.
6.Research progress of integrin in tissue repair
Lei CUI ; Chang LIU ; Yanchao JI ; Jinsong LI ; Song XUE ; Long JIANG
International Journal of Surgery 2013;40(10):674-677
As a transmembrane glycoprotein receptor family,integrin family mainly mediats cell-matrix interactions,participates in the function,through the unique bi-directional signal transduction pathways,played an important role in tissue repair process.The purpose of this article is to summarize the role of integrin family in the repair of tissue damage and sum up the current research progress.
7.Efficacies of total gastrectomy and proximal gastrectomy for the treatment of upper gastric cancer: a Meta analysis
Junsong LIU ; Xiangming CHE ; Duowei LI ; Houlong LONG ; Shicai HE ; Rui CHEN ; Shuai CHANG
Chinese Journal of Digestive Surgery 2013;(7):496-499
Objective To compare the efficacies of total gastrectomy (TG) and proximal gastrectomy (PG) for patients with upper gastric cancer.Methods Databases including Medline,Cochrane Library,Web of Science,China National Knowledge Infrastructure,Wanfang database were searched to retrieve literatures on surgical treatment of upper gastric cancer which were published from January 1980 to October 2011.According to different surgical procedures,all the patients were divided into PG group and TG group.Meta analysis were performed by RevMan 5.1.Categorical variables were presented by odds ratio (OR) and 95% confidence interval (95%CI).Results Thirteen literatures including 2622 patients with upper gastric cancer were retrieved.There were 1464 patients in the TG group and 1158 patients in the PG group.There was no significant difference in the 1-year survival rate between the 2 groups (OR =1.23,P > 0.05).The 3-and 5-year survival rates of patients in the TG group were significantly higher than those of the PG group (OR =1.74,1.45,P < 0.05).There were no significant difference in the 5-year survival rates of patients in TNM Ⅰ,Ⅱ,Ⅳ stages between the 2 groups (OR =0.94,1.31,2.03,P > 0.05),while the 5-year survival rate of patients in TNM Ⅲ stage of TG group was significantly higher than PG group (OR =2.29,P < 0.05) The overall recurrence rate of TG group was slightly lower than that of PG group,with no significant difference OR =0.44,P > 0.05).The local recurrence rate of TG group was significantly lower than that of PG group (OR =0.29,P < 0.05).There was no significant difference in the distal recurrence rate between the 2 groups (OR =0.60,P > 0.05).Conclusions The medium and longterm efficacies of TG are superior than that of PG.The stage of cancer should be taken into account to determine the plan of individual treatment.
8.Modified expression of extracellular signal-regulated protein kinase signal transducfion in rat braing and changed capacity of learning and memory of rats with chronic fluorosis
Yan-jie, LIU ; Qin, GAO ; Chang-xue, WU ; Yi-guo, LONG ; Zhi-zhong, GUAN
Chinese Journal of Endemiology 2009;28(1):32-35
Objective To investigate the expression of extraeellular signal-regulated protein kinase (ERK1/2)pathway in rat brains with fluorosis and the effects of fluoride on learning and memory of the rats,and to reveal the mechanisms of damaged nervous system resulted from the toxicity of the ion.Methods Seventy-two SD rats were divided into 3 groups and 24 rats were in each group.Three groups were fed respectively with different concentrations of fluoride(NaF)for 6 months to establish rat models with fluorosis.Controls were fed with tap water (NaF<0.5 mg/L):lower and higher concentration group were fed with water containing NaF(5,50 ms/L).Animals are sacrificed after 6 months of treatment with fluoride and the dissected brains were kept for analysis.The protein levels of ERK1/2 in rat brains were detected by Western-blotting and the mRNA level by RT-PCR. The spatial learning and memorizing ability was measured by Morris water maze test. Results The ERK1/2 protein in control group,lower and higher concentration group was 0.944±0.10,1.253±0.02,1.953±0.07,the differece being statistieally sighificant between any two groups (P < 0.05). The phospho-ERKl/2 protein in control group,lower and higher concentration group was 0.73±0.08,0.77±0.07,1.28±0.11,the differece being statistieally sighificant between any two groups(P < 0.05);the activation rate of phospho-ERK1/2 in lower and higher concentration group [(68.4± 3.8)%,(64.1±3.2)%] was decreased compared to control group[ (82.3±10.7)%],the differece being significant(P < 0.05). In the navigation trial,longer escape latencies of lower concentration group on the second, the third,the fifth and the sixth day were observed[ (46.0±8.0),(24.0±2.7),(8.9±5.3),(7.4±4.1 )s] compared to the control[ (39.3±6.9),(19.1±9.1 ),(8.3±3.4),(4.8±2.7)s],the differece being significant (P < 0.05 or < 0.01 );the similar results were also observed in the higher concentration group[ (36.9±16.8),(37.7±12.9), (19.7±7.6),(12.2±5.7 )s],and the escape latencies of the higher concentration group on the third,the fifth and the sixth day were longer than that in lower concentration group. In the probe test,the rats took more time to reach the first cross in lower and higher concentration group[(1.17±0.75),(4.18±1.10)s] than control group[ (5.89± 0.56 ) s ],the differece being significant (P < 0.05 or < 0.01 ) ;stayed shorter [ ( 17.05±4.25 ),(18.20±4.57 ) s ] than control [(25.37±5.65 )s ] in platform area (P < 0.01 );the activation rates of ERK1/2 were directly correlated with the time taken to reach the first cross platform located in the probe test(r = 0.364,P < 0.05) and the activation rates were also directly correlated with the escape latencies on the sixth day(r = 0.497,P < 0.05). Conclusion Long-term exposure of excessive fluoride induces the change of expression and activating rate of the ERK1/2 in rat brains,leading to the decreased capacity of learning and memory.
9.Level of oxidative stress in rat brains and learning and memory function of rats with chronic fluorosis
Qin, GAO ; Yan-jie, LIU ; Chang-xue, WU ; Yi-guo, LONG ; Zhi-zhong, GUAN
Chinese Journal of Endemiology 2008;27(4):371-373
Objective To investigate the changes of oxidative stress level in brain tissues and serum, and learning and memory in rats with oxidative stress level in nerve damage in chronic fluorosis. Methods The rats were randomly divided into 3 groups according to the body weight, eight rats in each group, i.e., control group, drinking water containing less than 0.5 mg/L of fluoride; lower fluoride exposure group, drinking water containing 5 mg/L of fluoride; higher fluoride exposure group, drinking water containing 50 mg/L of fluoride. The animals were examined six months after initiating the experiment. The total antioxidant capacity (T-AOC) and malondialdehyde (MDA), as well as learning and memory, were measured. Results Escape latency in higher fluoride exposed group[ (14.37±3.48)s] was significantly higher than that of controls[ (5.84±1.87)s] and exposed te lower fluoride [ (7.18±1.42)s], the difference being statistically signifieant(P<0.05). As compared with controls[ (2.17±0.11)× 103 U/L , (0.79±0.11)×103 U/g Pr] ,the rats exposed to higher fluoride and lower fluoride exhibited lower levels of T-AOC [(1.37±0.27)×103 U/L,(0.24±0.06)×103 U/g Prand (1.20±0.14) x 103 U/L,(0.41 ~ 0.10)×103 U/g Pr], the difference being statistically signifieant(P<0.05). As compared with controls[ (2.34±0.16) mmoL/L, (2.97±0.11)mmol/g Pr] and low fluoride exposed group[ (2.68±0.33)mmoL/L, (3.38±0.21)mmol/g Pr], higher level of MDA were observed in higher fluoride exposed group[ (3.72±0.59)retool/L, (4.01±0.21)mmol/g Pr], the difference being statistically significant(P<0.05). Conclusion The results indicated that higher amount of fluoride induced an increased level of oxidation, which might result in the decreased capacity of intelligence of rats with fluorosis.
10.Expression and clinical correlation of miR-141 in peripheral blood and tumor tissue in elderly patients with rectal cancer
Li FENG ; Yalei LV ; Liang CHANG ; Yan LIU ; Long WANG ; Zhihong XU ; Guiying WANG
Chongqing Medicine 2017;46(24):3339-3342
Objective To study the expression and clinical correlation of miR-141 in peripheral blood and tumor tissue in elderly patients with rectal cancer.Methods Rectal tumor tissue and tumor-adjacent normal tissues were taken in 75 cases of rectal cancer.The expression levels of miR-141 in peripheral blood and tumor tissue as well as tumor-adjacent normal tissues were determined by real-time polymerase chain reaction method.The correlation of miR-141 expression between peripheral blood and tumor tissue,and the correlation of peripheral blood miR-141 with clinicopathologic features and clinical prognosis were analyzed.Results (1) Compared with peripheral blood in the subjects undergoing normal physical examination or tumor-adjacent normal tissue in the patients with rectal cancer,the miR-141 expression level in peripheral blood and tumor tissue in the patients with rectal cancer was decreased significantly (P<0.01).(2) The cancer tissue miR-141 level in the patients with lymph node metastasis was positively correlated with peripheral blood miR-141 level (r=0.694,P<0.01),and cancer tissue miR-141 level in the patients without lymph node metastasis was positively correlated with peripheral blood miR-141 level (r=0.725,P<0.01).(3) The differences between peripheral blood miR-141 level with tumor stage,tumor differentiation degree and lymphatic metastasis had statistical significance (P<0.01).(4)The postoperative 6-month follow up displayed that among 75 cases,13 cases(17.33%) appeared replase/ metastasis,and peripheral blood miR-141 level was (2.64±0.34),which was significantly lower than that in the patents without replase/metastasis (P<0.01).Conclusion Expression variation trend of miR-141 in peripheral blood is similar to that in tumor tissue,which can reflect the clinicopathologic feature in the patients with rectal patients.