Objective To investigate the fermentation conditions for the heat-labile enterotoxin B subunit (LTB) of genetic recombinant E.coli. Methods The effects of different kinds of media, the range of pH, the induction time, the glucose concentration, and the fed-batch on LTB level in 10-liter fermenter under the condition of cascade controlling dissolved oxygen content were analyzed. Results Under the established conditions, the cell output per liter was 40.5g, and the expression level was 30.6%. Conclusion The results indicate that the stable fermentation technology with high efficiency has been established.

Objective To evaluate the efficacy and adverse events of irinotecan (CPT-11) combined with 5-FU in treatment of patients with advanced rectal cancer. Methods 86 patients were randomly divided into FOLFIRI treatment group and IFL treatment group. The efficacy and adverse events were observed. Results There was no statistically significant difference between FOLFIRI treatment group and IFL treatment group in CR、PR、SD and RR(all P ＞ 0. 05). There was also no statistically significant difference between FOLFIRI treatment group and IFL treatment group in adverse events(all P ＞ 0.05). Conclusion Irinotecan in combination with 5-FU was effective and had some tolerable adverse events in treating patients with advanced rectal cancer in both FOLFIRI treatment group and IFL treatment group.

Objective To investigate the prevalence rates of healthcare-associated infection (HAI)and antimicrobial use in a hospital in 2011-2013,and provide scientific basis for prevention and control of HAI.Methods HAI prevalence rates and antimicrobial use in a hospital in 2011-2013 were investigated by combination of bedside visiting and medical records reviewing.Results A total of 3 011 patients were investigated during three years,the prevalence rates were 3.49%, 2.87% and 3.98% respectively,difference of prevalence rates during three years were not significantly different (χ2 =2.105,P =0.356).Among HAI sites,lower respiratory tract ranked first,the major pathogens were gram-negative bacil-li,constituent ratios of different pathogens were not significantly different among three years(χ2 =1.003,P =0.972);anti-microbial usage rates and specimen detection rates among 3 years were both significantly different(χ2 =12.569,P <0.01;χ2=6.758,P <0.01,respectively),antimicrobial usage rate was highest in 2011(63.40%),and specimen detection rate was highest in 2012(62.14%).Conclusion Point prevalence rate in this hospital is at average national level,antimicrobial usage rate decreased, the consciousness of pathogenic detection gradually enhanced,clinical application management of antimicrobial agents still needs to be strengthened continuously.

The Third Military Medical University promotes the scientific research ability of the eight-year clinical medicine students through accurately fixing cultivation objectives,reasonably arranging teaching,implementing supervisor group for many-to-one teaching,carrying out plentiful innovative re-search projects,establishing reading system of scientific research papers,setting up scientific research innovation courses and setting up the credit system for innovative education. Eight-year program is signifi-cantly better than the five-year program in cultivating abilities of problem-discovering,researching and thesis writing.

Acupuncture therapy is a therapeutic method in traditional Chinese medicine. Its clinical efficacy has widely accepted internationally but its mechanism of action is still unclear. In recent years, more and more researchers began to use brain network analysis to explore the mechanism of action of acupuncture. This article reviews the significance of brain network analysis in the study of acupuncture effect, that is, brain network analysis can effectively assess changes in cerebral function in chronic pain and observe the real therapeutic effect of acupuncture. It also reviews various methods of brain network analysis, including brain functional connectivity (FC) analysis, amplitude of low frequency fluctuations (ALFF) analysis, regional homogeneity (ReHo) analysis, small-world network (SWN) analysis, positron emission computed tomography (PET) and diffusion tensor imaging (DTI); the shortcomings and prospects of brain network analysis in the application of acupuncture. A summary of the newest research advances in the application of brain network analysis to the study of acupuncture effect provides a certain reference for the future scientific study.

Objective To explore the effect of acupuncture at Yanglingquan (GB 34) on white matter structure of stroke patients. Methods 6 stroke patients with left hemiplegia in recovery stage were recruited. The sham acupoint was as control, the diffusion tensor imaging (DTI) data in a 1.5 T SIEMENS scanner before and after acupuncture at Yanglingquan point was collected. Differences between the two conditions were compared with the Tract-Based Spatial Statistics (TBSS), which is one of FSL tools. Results The brain areas included left precentral gyrus, postcentral gyrus, middle temporal gyrus, inferior temporal gyrus, supplementary motor area, inferior frontal gyrus, internal capsule, fornix and bilateral inferior parietal gyrus, in which the verum acupuncture had stronger effect on fractional anisotropy (FA) than the sham acupuncture. White matter tract such as pyramidal tract, arcuate fasciculus, corpus callosum and other inter-cortex association fibers passed these brain areas. The fiber tracts and brain areas were found to be involved with multiple brain networks of motor compensation, somatosensory, language and cognition. Conclusion Compared with the sham point, acupuncture at Yanglingquan point can change the microstructure of stroke patients' white matter, mainly in the inaffected side and related to multiple functional networks.

AIM:ToinvestigatetheeffectofP21-activatedkinase1(PAK1)andlymphoidenhancer-binding factor 1(LEF1) on the proliferation of esophagus cancer cells .METHODS:Real-time PCR was applied to detect the mR-NA expression of PAK1 and LEF1 in the esophagus cancer tissues .MTT assay were used to measure the proliferation of hu-man esophagus cancer cell line KYSE transfected with PAK 1 and LEF1.RESULTS: The mRNA expression of PAK1 in the esophagus cancer tissues was lower than that in control group (P<0.05).The mRNA expression of LEF1 and tran-scription factor 4 (TCF4) in the esophagus cancer tissues was higher than that in control group (P<0.05).The prolifera-tion of KYSE cells with over-expression of PAK1 and LEF1 was higher than that in control group .No significant change of apoptosis between the KYSE cells with over-expression of PAK1 and LEF1 and control group was observed .CONCLU-SION:The expression of PAK1 decreases and the expression of LEF 1 increases in esophagus cancer tissues .LEF1 domi-nantly regulates the proliferation of esophagus cancer cells .

Objective To investigate protective effects of thrombopoietin ( TPO) on cerebral model control in rats and associated signal transduction pathway. Methods Thread embolism was performed to generate cerebral ischemia-reperfusion rat model. Eighty male SD rats were randomly divided into sham operation group, model control group, TPO group, TPO and Janus kinase 2 ( JAK2 ) kinase inhibitor ( AG490 ) group. Before 30 min of ischemia-reperfusion, TPO group was given TPO (5 μg·kg-1) by intraperitoneal injection, TPO + AG490 group was given TPO (5 μg·kg-1) before 30 min of ischemia reperfusion, then given AG490 (8 μg·kg-1), and model control group were given the same dose of 0. 9% sodium chloride solution. The observation time points were 6, 12, 24, and 48 h after ischemia reperfusion. Immunohistochemical staining and Western blotting were used to measure the protein levels of Bcl-2, JAK2 and signal transducer & activator of transcription (STAT3). TdT-mediated dUTP nick end labeling (TUNEL) was used to detect apoptosis. Results Compared with model control group, the number of apoptotic cells were significantly reduced [(67. 50±9. 37) vs. (40. 20±7. 47)], the expression levels of Bcl-2, JAK2 and STAT3 protein were significantly increased [(35. 40±7. 39) vs. (78. 70±9. 75);(35. 68±6. 75) vs. (62.35±7.53); (25.40±9.45) vs.(55.36±9.69), respectively] 24 h after ischeia reperfusion in the TPO group (all P<0. 05). Compared with the TPO group, the Bcl-2, JAK2 and STAT3 protein levels were significantly decreased in TPO and AG490 group [(78. 70±9. 75) vs. (55. 40±9. 35);(62. 35±7. 53) vs. (40. 68±5. 89); (55. 36±9. 69) vs. (30. 40±9. 39), respetively], and the number of apoptotic cells was significantly increased [(40. 20±7. 47) vs. (55. 23±7. 65)] (all P<0. 05). Conclusion TPO can inhibit cell apoptosis after ischemia-reperfusion injury, the mechanism might be related to the activation of JAK2/STAT3 signal transduction pathway through raising the expression of Bcl-2 gene.

Objective To investigate the expression of Kuppel-like factor 2( KLF2 )after focal cerebral ischemia-reperfusion( I/R)injury in rats and the intervention effect of nuclear factor kappa B ( NF-κB)inhibitor. Methods Sixty healthy male SD rats were randomly divided into a sham operation group,an I/R group,and a NF-κB inhibitor group( n=20 in each group). A focal cerebral I/R model was induced by the intraluminal suture method,and NF-κB inhibitor( pyrrolidinedithio carbamate,PDTC)was given to intervene. The observation time points were 6,12,24,and 48 hours after I/R. Reverse transcription-polymerase chain reaction(PCR)and Western blot were used to measure KLF2 mRNA and protein expression in ischemic brain tissue. Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of serum tumor necrosis factorα( TNF-α),and they were compared among groups. Results Compared with the sham operation group,the expression levels of KLF2 mRNA and protein in I/R group in the ischemic brain tissue at each time point were averagely decreased( the relative expression levels of KLF2 mRNA:0. 46 ± 0. 03 vs. 0. 82 ± 0. 04,0. 30 ± 0. 04 vs. 0. 78 ± 0. 05,0. 18 ± 0. 04 vs. 0. 76 ± 0. 02,0. 26 ± 0. 02 vs. 0. 81 ± 0. 04,respectively;the relative expression levels of KLF2 protein:0. 46 ± 0. 04 vs. 0. 80 ± 0. 02,0. 30 ± 0. 02 vs. 0. 79 ± 0. 02,0. 15 ± 0. 02 vs. 0. 77 ± 0. 01,0. 24 ± 0. 01 vs. 0. 79 ± 0. 02,respectively). They reached the lowest values at 24 hours after I/R,while the serum TNF-αlevels were increased. There were significant differences(all P<0. 05). After giving NF-κB inhibitor PDTC,the expression levels of KLF2 mRNA and protein at 6,12,24,and 48 hours after I/R were upregulated differently compared with the I/R group. The relative expression levels of KLF2 mRNA were 0. 61 ± 0. 04,0. 44 ± 0. 03,0. 34 ± 0. 02,and 0. 43 ± 0. 04, respectively. Those of KLF2 protein were 0. 60 ± 0. 02,0. 43 ± 0. 02,0. 33 ± 0. 01,and 0. 44 ± 0. 03, respectively,while the levels of TNF-αwere decreased. There were significant differences(all P<0. 05). There was a negative correlation between the KLF2 mRNA levels and the serum TNF-αlevels at each time point in the I/R group and the PDTC group( r= —0. 728 ,P<0. 05 ). Conclusions The expression levels of KLF2 mRNA in brain tissue are decreased after I/R,and it is negatively correlated with the serum TNF-α levels. It may be involved in the pathological process of I/R by NF-κB pathway mediated inflammatory reaction.

Objective To study the effect of anterolateral calf flap with extra-territorial blood supply in repairing large area of foot skin tissue injury. Methods 40 cases of arge area of foot skin tissue injury were divided into the observation group which were given anterolateral crural flap transplantation and the control group which were given anterolateral thigh flap transplantation. The wound healing condition and walking ability were observed. Results The level of wound healing, Holden walking ability and FAC walking ability were better than control group (P<0. 05). In the observation group, wound healing time was (15. 2 ± 6. 2) d and donor skin crusting time was (9. 4 ± 1. 5) d, which were less than the control group(P<0. 05). Conclusion Anterolateral calf flap with extra-territorial blood supply has positive clinical value as it can improve wound healing and walking function.