Objective To study the safety and efficacy of intraoperative implanting slow-released 5-Fu after hepatectomy in patients with hepatocellular carcinoma (HCC). Methods 59 HCC patients undergoing hepatectomy from January 2008 to January 2009 were divided into two groups:to receive slowrelaesed 5-Fu in treatment group and nothing in control group.Postoperatively no patients in both group took chemotherapy.The serous value of WBC,ALT,AST,TBIL and AFP were measured in all cases on 1 day before and 3 weeks after the operation.During the first six months,AFP was measured and imaging studies were done one month after discharge,and every 3 months since then.CT-guided biopsy was used to confirm the recurrence of HCC. Disease-free survival rate and overall survival rate in two groups calculated respectively 6-,12-,18- and 24-months after the operation. Results The serous value of WBC,ALT,AST,TBIL was not different between treatment group and control group 3 weeks after the operation (separately t =0.801,- 0.854,- 1.948,- 0.503,all P ＞ 0.05).AFP was (361.58 ± 431.06) μg/L,( 17.02 ± 15.55 ) μg/L,(43.61 ± 58.03 ) μg/L in treatment group and (495.50 ± 441.63 ) μg/L,(26.82 ±60.46) μg/L,(127.48 ±229.79) μg/L in control group preoperatively,3 weeks and 6 months after the operation.AFP was lower in treatment group than that in control group ( t =- 2.065,P ＜ 0.05 ).The disease-free survival rate in treatment group was 95.8％,91.7％,79.2％,75.0％ at 6-,12-,18- and 24-months after the operation,in control group it was 94.3％,71.4％,60.0％,48.6％ ( x2 =4.035,P ＜0.05).The overall survival rate was respectively 100％,95.8％,91.7％,83.3％ in treatment group and was 100％,94.3％,77.1％,60.0％ in control group( x2 =3.931,P ＜0.05).Conclusions Intraoperativeimplanting slow-released 5-Fu during the hepatectomy was safe and effective method to reduce the recurrence rate of hepatocellular carcinoma and prolongs postoperative disease free and overall survival in HCC patients undergoing hepatectomy.

Objective:To investigate the immuno-effects triggered by dendritic cells (DCs) pulsed with Heat-treated lymphoma cells.Methods:Mononuclear cells from healthy human peripheral blood(PBMC) were isolated and DCs were generated in vitro with normal methods.Lymphoma cells were heated (42℃,2 h) and then loaded onto DCs.Immunotypes of DCs and lymphocytes were detected with flow-cytometric analysis.IFN-γ releasing was detected by ELISPOT,lymphocyte cytotoxicity was analyzed by MTT method after mixing with DC reaction.Results:After pulsing DCs with hyperthermia lymphoma cells,CD80~+,CD86~+ and HLA-DR existed higher than those of the control group,IFN-γ releasing and cytotoxicities were also strengthened,but there was no difference between them,cytotoxitic lymphocytes (CTLs) were produced more as CD8~+,CD25~+ and CD56~+ cells by flow-cytometric analysis.Conclusion:Anti-lymphoma immuno-effects could be strengthened by loading DC with hyperthermia lymphoma cells.

Objective To investigate whether TNF-α can promote, but TGF-β can reduce Gli2 expression in primary leukemia cells. Methods (1)Western blot was used to detect Gli2 protein expression in leukemic cells. (2)Primary leukemia cells were treated with TGF-β1 or TNF-α,and expressions of Gli2 and TGF-βR were detected. (3)Primary leukemia cells were treated with TGF-β1,TNF-α or/and SIS3, and Gli2 protein expression was detected. Results (1)The primary leukemia cells could express Gli2 protein. (2)Significant reduction of Gli2 mRNA was observed in 1 ~ 10 ng/mL TGF-β1-treated primary leukemia cells for 12 ~ 24 h, No significant differences of Gli2 mRNA was found between the control group and TNF-α group. (3)Protein expresson of TGF-βRⅠand TGF-βRⅡ in 5 ng/mL TNF-α-treated primary leukemia cells for 24 h was higher than that of the control group. (4)Gli2 portein expression was lower in TGF-β1 with or without TNF-α-treated primary leukemia cells for 24 h than that in the control group, and Gli2 portein expression in the TGF-β1 plus TNF-α group was lower than that in TGF-β1 group. Conclusion Combinations of TGF-β and TNF-α can efficiently inhibit Gli2 expression than TGF-β alone in primary leukemia cells.

Objective To screen and identify the predominant T-and B-cell (T-B) combined antigenic epitopes in outer membrane protein GroEL of Helicobacter pylori (H.pylori).Methods The entire groEL gene of H.pylori strain NCTC11637 was amplified by PCR,and then a prokaryotic expression system for groEL gene was constructed.The expressed target recombinant protein rGroEL was analyzed by SDSPAGE and purified by Ni-NTA affinity chromatography.Laser confocal microscopy was applied to determine the distribution of GroEL in H.pyloristrains of NCTC11637 and SS1.The outer membrane protein (OMP) samples were extracted from the two strains using Triton X-114 method.GroEL in OMP samples was detected by Western blot assay.Special bioinformatic softwares were used to predict T-B combined antigenic epitopes on GroEL molecule,and phage display systems for every T-B combined antigenic epitope were established.Western blot assay and ELISA were used to detect the immunoreactivity of recombinant T-B combined antigenic epitope-containing phage PⅢ proteins and artificially synthesized T-B combined antigenic epitope peptides,respectively.Results The groEL genes from H.pylori strains of NCTC11637 and SS1 showed 97.68％ ～99.63％ identities in their nucleotide and amino acid sequences.The established prokaryotic expression system could efficiently express soluble rGroEL.GroEL located on the outer membrane of H.pylori strains of NCTC11637 and SS1.Among the six major predicted T-B combined epitopes (GroEL168,GroEL258,GroEL288,GroEL365,GroEL396 and GroEL438),GroEL168 showed the strongest positive immunoblotting signal.The results of ELISA showed that the immunoreactivity of GroEL168 was also the strongest (P＜0.01),followed by GroEL258 and GroEL288 (P＜0.05).Conclusion GroEL is an outer membrane protein of H.pylori.GroEL168 being the predominant T-B combined antigenic epitope of GroEL can be used to develop multiple antigenic peptide vaccines of H.pylori.

To identify the species of Mycobacterium clinical isolates by molecular biology techniques,six clinical isolates which were preliminarily recognized as Mycobacterium tuberculosis by the TCH and PNB culture methods were selected in this study.PCR was applied to amplify the oxyR-ahpC interval resistant-zone(intergenic region) and the length of PCR product in four strains was the same as that of H37Rv,while the length in the other two strains was different from that of H37Rv but same as Mycobacterium intracellulare 95002.With sequencing and on-line homology comparison with H37Rv,U18263 and U71061,the DNA sequence of oxyR-ahpC intergenic region displayed a 99% homology with Mycobacterium intracellulare U71061 and an 84% homology with Mycobacterium tuberculosis H37Rv.In addition,the results of hsp65 PCR-restriction fragment length polymorphism analysis and multi-locus PCR amplification in the two strains were identical with those of Mycobacterium intracellulare 95002.These two clinical isolates which were preliminarily recognized as Mycobacterium tuberculosis by PNB and TCH culture methods were finally identified as Mycobacterium intracellulare.Results predicted that the application associated with various techniques of molecular biology would provide a faster,easier and more correct way for the species identification of Mycobacterium.

Nocardia is a genus of gram-positive,weakly acid-fast,filamentous aerobic actinomycetes,which mainly causes infection in immunocompromised persons. We reported a successfully treated fatal case of severe pneumonia caused by Nocardia farcinica in a hospital,then reviewed 25 domestic and abroad case reports about nocardiosis combined with severe pneumonia occurred since 2006,so as to improve health care workers'cognition on clinical manifestations,image features,pathogenic characteristics,and diagnostic and treatment schemes of se-vere pneumonia caused by Nocardia farcinica.

Objective:To evaluate the rehabilitation efficacy of early mobilization based on collaboration care model for patients with laparoscopic colorectal surgery.Methods:Cluster sampling method was used in the department to recruit colorectal cancer patients with laparoscopic surgery. The control group (49 cases) received routine perioperative care and exercise, and the intervention group (47 cases) received the coordinated early mobilization combined with routine perioperative care and exercise, from January to March 2019. Primary outcome were health status and the proportion of patients returning to preoperative functional walking capacity (6-min walk test) at 4 weeks after surgery. The in-hospital mobilization (time out-of-bed), time to achieve discharge criteria, time to recover gastrointestinal function and complication rate were explored.Results:In intervention group,89.4%(42/47) of patients achieved mobilization target on 4 days after surgery compared with 42.6%(20/47) on the day of surgery. Time out of bed were greater in the intervention group compared with the control group, and there were differences between the two groups( Z values were -8.437--7.381, P<0.01). Time to recover gastrointestinal function and the recovery of energy on 3 days after surgery were (58.74±17.41) h, (59.02±9.46) points in the observation group, and (71.82±21.53) h, (62.61±7.68) points in the control group, and there were significant differences between the two groups ( t values were -3.263, -2.046, P<0.05). But other outcome measures were not different between the two groups ( P>0.05). Conclusions:For colorectal laparoscopic surgery patients, the coordinated early mobilization improved the adherence to ambulation, in-hospital mobilization, time to recover gastrointestinal function and recovery of energy to promote rehabilitation.

Objective To study the effect of tumor size on the expression of adrenal cholesterol homeostasis mole -cules in H22 hepatoma-bearing mice.Methods Two hundred and twenty mice were injected with H 22 hepatoma cells to their right armpit.On the 11th day after injection, the mice were sorted according to the tumor size .18 mice with large tumor (large tumor group) and 18 mice with small tumor (small tumor group) were sacrificed, and the tumors were weighed.A control group consisting of 18 normal Kunming mice was also included in this study .The plasma TC, TG and HDL-C were detected using total cholesterol , triglycerides or HDL-C assay kits , respectively .The mRNA expressions of Srb1, Ldlr, Npc1, Npc2, Stard3, Hmgcr, Lipe, Acat1, Abca1, Abcg1, Srebp-1c, Lxrα, Lxrβ, Rxrα, Apoa1 and Apoe were tested by real-time quantitative RT-PCR ( qRT-PCR ) , and Gapdh and β-actin were used for normalization .SRB1 and ApoA1 proteins were analyzed by Western blot .Results The tumor weight was significantly higher in the large tumor group than that in the small tumor group (P<0.05).Compared with the control group , the plasma HDL-C was significant-ly decreased in the two hepatoma groups (P<0.05).The expression levels of Srb1, Ldlr, Apoa1 mRNA and SRB1 protein were significantly increased in the large tumor group (P<0.05 for all).The ApoA1 protein level was significantly higher in the large tumor group than that in the small tumor group (P<0.05).The expressions of Acat1, Lipe, Abca1 and Abcg1 mRNA were significantly lower in the large tumor group than those in the small tumor group (P<0.05 for all).However, the expressions of Srebp-1c, Lxrαand RxrαmRNA were not significantly changed , then, Srebp-1c, Lxrβand RxrαmRNA expressions were significantly up-regulated in the small tumor group (P<0.05).The expressions of Hmgcr and Apoe mR-NA were not significantly different in the two groups .Conclusions In hepatoma-bearing mice , due to the adaptation to tumor-induced chronic stress response , the adrenal cortical cells can effectively utilize intracellular cholesterol to synthetize cortical hormones .

Objective To analyze the effects of achievement motivation and family environ-ment of undergraduates majoring in preventive medicine on creativity tendency. Methods The self-administered questionnaire was used to survey the general characteristics , achievement motivation , family environment, and Williams creativity tendency of undergraduates (n=243) from grade 1 to grade 5 majoring in preventive medicine in a medical college. The effects of achievement motivation and family environment on creativity tendency were tested with multiple linear regression analysis. SPSS 17.0 software was used for statistical analysis. The indexes between boys and girls was compared with group t test, and single factor analysis of variance was made for comparison between grades, SNK method was used to test and compare two groups; linear correlation analysis was made to inspect the correlation between the two indicators, and multiple linear regression analysis method was used for further analysis. Results The survey recycled 226 copies of effective questionnaires. The scores of achievement motivation, family environment, and Williams creativity tendency and all dimen-sions had no statistically significant difference among the five grades, or between the male and female (P>0.05). The positive influence factors of creativity tendency and its dimensions were pursuing success, the in-formative property, the entertaining property and organizational property, the negative influence factor was avoiding failure. R2 was 0.094-0.265 (P=0.000-0.048). Conclusion Achievement motivation and family environment of undergraduates majoring in preventive medicine may have posi-tive effects on creativity tendency. Various comprehensive measures should be taken to improve family environment and enhance achievement motivation, and to promote to increase creativity tendency, and to cultivate students' creativity and improve students' scientific research ability.

Objective To study the application of implantable telemetry and whole-body plethysmography to observe the changes of circadian rhythm in conscious rats and evaluate the pharmacological safety of doxofylline, and to provide a basis for the future application of this technological system for drug safety evaluation.Methods Eight healthy SPF Sprague-Dawley rats were used in this study, 4 males and 4 females.The rats were implanted with telemetry transmitters by surgery to establish a telemetry system combined with plethysmography to observe the changes of 24 h physiological parameters and circadian rhythm in conscious rats at 14 d after operation, including heart rate (HR), blood pressure, the time interval from the Q wave to point A in the ECG of the aortic pressure wave (QA interval), respiration, activity, body temperature and pulmonary function parameters.The rats were divided into 3 groups: normal control group, doxofylline 40 mg/kg and 80 mg/kg groups, and the performance was validated by aerosolizing saline, doxofylline 40 mg/kg and 80 mg/kg inhalation, respectively, to observe the changes in physiological parameters after the drug administration.Results The physiological parameters of rats showed obvious changes in circadian rhythms at 14 d after operation.Compared with the normal control group, the doxofylline 40 mg/kg-treated group showed significantly increased changes of HR, tidal volume (TV), minute ventilation (MV), 50% expiratory flow (EF50), peak inspiratory flow (PIF) and peak expiratory flow (PEF) (P<0.01), significantly decreased respiratory frequency, QA interval and enhance pause (Penh) (P<0.05, P<0.01), but no significant differences in the blood pressure, activity and body temperature (P>0.05).Compared with the normal control group, the group treated with doxofylline 80 mg/kg had significantly increased HR, blood pressure, TV, MV, EF50, PIF and PEF (P<0.01), significantly decreased respiratory frequency, QA interval and Penh not (P<0.01), but not significantly changed activity and body temperature (P>0.05).Conclusions The application of implantable telemetry and whole-body plethysmography in this study does not obviously affect the circadian rhythm, and can sensitively monitor the relevant cardiovascular and respiratory parameters in conscious rats.It can be used in drug safety pharmacological research of cardiovascular and respiratory systems in conscious rats.