Objective To explore the effects of X-ray irradiation combined with RNAi against signal transducer and activator of transcription 3(STAT3)on the radiosensitivity of human esophageal carcinoma cells.Methods Human esophageal carcinoma cells of the line Eca-109 were euhured.Three pairs of DNA template aiming at the base sequences of the coding regions 2037-2055,1243-1261,and 455-473 of the STAT3 mRNA were synthesized(siRNAI,siRNA2,and siRNA3),and a negative sequence was synthesized to be used as control.STAT3-siRNA positive recombinant plasmids(pRNAT-U6.1-siRNAI,pRNAT-U6.1-siRNA2, and pRNAT-U6.1-siRNA3), and a STAT3-siRNA negative recombinant plasmid (pRNAT-U6.1-negative)were thus constructed and then transfected into the cultured Eca-109 cells,which were divided into transfection reagent control group,pRNAT-U6.1-siRNAl-3 transfection groups,and pRNAT-U6.1-negative centrel group.The positive eell clones were screened.RT-PCR and Westem blotting were used to detect the STAT3 mRNA and protein expression.The transfected Eca-109 cells were exposed to 0,2,4,6,and 8 Gy of X-rays,respectively,and the survival fraction of the cells was analyzed by clone formation assay.Flow cytometry was applied to analyze the cycle arrest and cell apoptosis 4 Gy post-irradiation.Results Agarose gel electrophoresis confirmed the successful construction of the plasmid pRNAT-U6.1-siRNA.RT-PCR and Western blotting demonstrated that the mRNA and protein expression levels of STAT3 transfected with sTAT3-siRNA3 were both significanfly lower than those of the control groups.At 2-8 Gy, the survival fractions of the siRNA3 group were aU significantly lowered than those of the control group(t＝-0.228--0.051,P<0.05).Flow cytometry showed that the percentage of the cell cycle G0/G1 phase and the apoptosis rate of the siRNA3 group were both significantly higher than those of the control groups at 4 Gy post-irradiation(t＝-13.137-16.350,P<0.01).Conclusions X-ray irradiation combined with RNAi against sTAT3 could inhibit the proliferation of the human esophageal carcinoma cells,induce cell cycle arrest and apoptosis,improve the radiosensitivity in Eta-109 cells.

BACKGROUND:Umbilical cord blood transplantation can ameliorate hepatic and immunologic function,repair hepatic injury,and promote hepatic regeneration,however,the differentiation mechanism and biological characteristics remain poorly understood,and the long-term efficiency need to be explored.OBJECTIVE:To investigate the long-term therapeutic efficacy of infusing umbilical cord blood and blood plasma in treating chronic severe hepatitis B patients.METHODS:Totally 50 chronic severe hepatitis B patients received treatment at the Second Xiangya Hospital,Central South University from January 2003 to January 2004 were randomly divided into the treatment and control groups,with 25 cases in each group.All patient were accepted an ordinary synthetic treatment,and the differences between age,pathogenetic condition,medication had no significance.The umbilical cord blood was obtained from healthy full-term spontaneous delivery parturient,centrifugated,remained karyotes and cord plasma,and used within 24 hours.Patients in the treatment group were received umbilical cord blood infusion,200 mL once,1 2 times per week,totally,each patients infused 4-8 times(mean 5 times);those in the control group were infused with adult fresh blood plasma.The changes of hemogram and hepatic function were measured.RESULTS AND CONCLUSION:All the patients were followed-up for 1 year.The hemogram and hepatic function indexes were similar in the 2 groups before treatment(P ＞ 0.05).The hemogram index had no obviously difference at 1 year after treatment (P ＞ 0.05),but the alanine aminotransferase and total bilirubin levels were decreased in the treatment compared with the control group(P ＜ 0.05),but the albumin was significantly increased(P ＜ 0.05).Compared with before treatment,the platelet level had no significant changes at 1 year after treatment,but the alanine aminotransferase and total bilirubin levels were deeply decreased(P＜ 0.05),albumin was significantly increased(P ＜ 0.05);the platelet and albumin levels were dramatically decreased in the control group(P ＜ 0.05).It suggested that umbilical cord blood infusion can improve the hepatic function and hemogram;therefore,it can be served as supplementary therapeutic measure for severe hepatitis.

Objective: To study the cell proliferation, cell cycle and apoptosis of esophageal carcinoma Eca-109 cells treated with RNA interference technique to silence signal transducers and activators of transcription 3 (STAT3) gene. Methods: Three pairs of DNA template coding siRNA specific for human STAT3 gene mRNA were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pRNAT-U6.1/neo plasmid to construct STAT3-siRNA expression vector which was then transfected into Eca-109 cells. The expression of STAT3 mRNA and protein in cancer cells was detected by RT-PCR and Western blot, respectively. The cell proliferation, cell cycle distribution and apoptosis were examined by MTT and flow cytometry. Results: STAT3-siRNA expression vector was successfully constructed and identified by sequencing. The results of RT-PCR and Western blot demonstrated that STAT3 expression in Eca-109 cells transfected with STAT3-siRNA expression vector was significantly higher than that in the control group (P＜ 0.01). MTT showed that after transfection of the siRNA vector into Eca-109 cells, cell proliferation was obviously reduced and the cell growth inhibition ratio in the siRNA3 group was 35.68%, significantly higher than that in the control group (P＜0.01). Flow cytometry results suggested that cell cycle arrest and more apoptosis were observed in the siRNA3 group. Cell cycle was arrested at G_0/G_1 phase, and the rate of apoptosis was 13.26%, much higher than that in the control group (P＜0.01). Conclusion: Silencing STAT3 gene by RNA interference technique can effectively inhibit STAT3 expression, suppress the proliferation of Eca-109 cells, induce cell cycle arrest at G_0/G_1 phase, and promote apoptosis.

Background and purpose:Several reports demonstrated that the expression of STAT3 has been found to be an oncogene in solid tumors such as head and neck squamous cell carcinoma,esophageal carcinoma and prostate carcinoma.This study was done to explore the effects of X-ray irradiation combined with RNAi against STAT3 on proliferation and apoptosis of human esophageal carcinoma cell line Eca-109.Methods:Three pairs of DNA template coding siRNA were synthesized against STAT3 to reconstruct pRNAT-U6.1-siRNA-STAT3,which was transfected into Eca-109 cells,the positive cell clones were screened with G418.Inhibitory effect of STAT3 mRNA and protein in Eca-109 cells was detected by RT-PCR and Western blot,respectively.The transfected cells were exposed to 0,2,4,6 and 8 Gy X-ray respectively;the survival fraction of Eca-109 cells was analyzed by clone formation assay,and flow cytometry was applied to analyze cell apoptosis at the dose of 4 Gy.Results:pRNAT-U6.1-siRNA-STAT3 was reconstructed and identifi ed as correct by sequencing.RT-PCR and Western blot analyses demonstrated that STAT3-siRNA could obviously reduce the expression of STAT3 mRNA and protein in Eca-109 cells.Clone formation assay and flow cytometry results showed that irradiation at different doses combined with STAT3-siRNA could inhibit the proliferation of Eca-109 cells,irradiation with 4 Gy X-ray could induce apoptosis.Conclusion:X-ray irradiation combined with RNAi against STAT3 could inhibit the proliferation of Eca-109 cells and induce apoptosis.

Objective To explore the influencing factors of routine blood test results in the laboratory ,to ensure the accuracy of test results .Methods The influence of the different area of drawing blood and different testing time after blood collection were ob-served .Results The leukocyte count ,red blood cell count ,hemoglobin level of finger blood were obviously higher than those of venous blood ,and the platelet count was lower than that of venous blood ,the difference was statistically significant(P<0 .05) .≥6 h test specimens of blood platelet count was significantly lower ,compared to immediate detection and less than 6 h ,the difference was statistically significant(P<0 .05) .Conclusion The improvement of the system ,operation specification ,improving the quality of samples are helpful to improve the accuracy of test results .

Objective To investigate the expression of phosphorylated protein kinase B1 (pAkt1) and human telomerase reverse transcriptase (hTERT) in ovarian epithelial carcinoma (EOC) and its relationship with prognosis, and to analyze the correlation between pAkt1 and hTERT expression. Methods 92 patients with EOC in Tangshan Gongren Hospital from January 2012 to December 2016 were selected. The expressions of hTERT and pAkt1 proteins were detected by immunohistochemistry. The relationship between the expressions of pAkt1 and hTERT proteins and their relationship with clinical prognosis were analyzed. Results Of the 92 patients with EOC, 68 cases (73.9 %) had positive expression of pAkt and 52 cases (56.5 %) had positive expression of hTERT. There was no significant correlation between expressions of pAkt1 and hTERT proteins in EOC (r= 0.284, P= 0.633). The expressions of pAkt1 and hTERT proteins were not related with age, tumor pathology type and the International Federation of Gynecology and Obstetrics (FIGO) staging (all P> 0.05), but had significant association with tumor pathology differentiation (χ2= 2.694, P= 0.005; χ2=2.284, P=0.018). The disease-free survival of patients with both pAkt1 and hTERT positive was shorter than that of the other groups (P= 0.013). Conclusion The prognosis of EOC patients with high expression of pAkt1 and hTERT proteins is poor.

[ ABSTRACT ] AIM: To investigate the expression and clinical significance of bone morphogenetic protein 3 (BMP3) in hilar cholangiocarcinoma tissues.METHODS: Thirty cases of hilar cholangiocarcinoma specimens were col-lected.The expression of BMP3 at mRNA and protein levels in the tumor tissues and paracancerous tissues was detected by real-time PCR and Western blot.The hilar cholangiocarcinoma paraffin-embedded specimens (n=103) were collected. The protein expression of BMP3 was determined by immunohistochemical method, and the relationship of BMP3 protein ex-pression with clinical pathological characteristics was evaluated.RESULTS:In the 30 patients with hilar cholangiocarcino-ma, the expressions of BMP3 protein and mRNA in 22 cases of tumor tissues were significantly decreased compared with the adjacent normal tissues.The results of immunohistochemistry showed that 87 cases were negative and 16 cases were weakly positive in all 103 cases of hilar cholangiocarcinoma.The expression of BMP3 protein was associated with the tumor TNM staging, lymph node metastasis and tumor differentiation (P<0.05).CONCLUSION:BMP3 gene might be inhibited in human hilar cholangiocarcinoma.The down-regulation of BMP3 gene might be associated with the carcinogenesis and devel-opment of hilar cholangiocarcinoma.

Objective To provide a scientific basis for nursing managers to promote the professional quality of life of nurses.Methods Totally 373 nurses from three general hospitals in Jinan by convenience sampling technique were recruited and assessed with the Professional Quality of Life Scale Version 5 (ProQOL-V) and the Five Facet Mindfulness Questionnaire.The relationship between professional quality of life and mindfulness was analyzed.Results The average score of compassion satisfaction was (29.71±6.61) points,the average score of compassion fatigue was (52.82±8.04) points,which were in the middle level of them.Compassion satisfaction was correlated with mindfulness,observation,description,non-reactiveness and non-judgment (P ＜ 0.05).Compassion fatigue was correlated with mindfulness,observation,awareness of the action,non-reactiveness and non-judgment (P ＜ 0.05).Multiple regression analysis showed that the three dimensions (observation,non-reactiveness and non-judgment) of mindfulness could predict 15％ variance of compassion satisfaction and the three dimensions (observation,awareness of the action and non-judgment) of mindfulness could predict 34.1％ variance of compassion fatigue after controlling demographic variables.Conclusions Mindfulness can effectively predict nurses' professional quality of life.Nursing managers should take some mindfulness training in order to improve their professional quality of life.

[ ABSTRACT] AIM:To investigate the effects of microRNA145 ( miRNA145 ) on the viability, apoptosis, inva-sion and metastasis of hepatoma HepG2 cells.METHODS: HepG2 cells were randomly allocated into 3 groups: blank control group, empty mimic transfected group and miRNA145 mimic transfected group.Under the induction of Lipofectami-neTM 2000, the recombinant was transfected into HepG2 cells.After transfection, the expression level of miRNA145 was detected by real-time PCR.The protein level of N-cadherin and the mRNA expression levels of miRNA145 and N-cadherin were detected by Western blot and real-time PCR.The cell viability was detected by MTS assay.The cell cycle and apopto-sis were analyzed by flow cytometry.Invasion and metastasis were detected by Transwell assay.RESULTS:Compared with negative control, miRNA145 expression was up-regulated significantly, while the expression of N-cadherin was down-regu-lated significantly.Meanwhile, the cell viability, cell cycle, apoptosis, invasion and metastasis of hepatoma HepG2 cells were all significantly inhibited (P<0.05).CONCLUSION:miRNA145 dramatically inhibits viability, apoptosis, inva-sion and metastasis of hepatoma cells.