BACKGROUND:Numerous studies have demonstrated that mild hypothermia has a better protective effect on neurons after cerebral infarction.
OBJECTIVE:To investigate theeffect of mild hypothermia on nerve regeneration microenvironment of infarcted area in rat models of cerebral infarction and analyze its possible effects on neural functional recovery after cerebral infarction.
METHODS:Twenty out of 65 adult femaleSprague-Dawleyrats were randomly selected as the sham group. The remaining 45 rats were subjected to carotid artery ligation to establish rat models of cerebral infarction. Five rats were rejected because of modeling failureor death, the remaining 40 rats were randomly and evenly divided into cerebralinfarction and mild hypothermia groups. The head temperature of rats in the cerebral infarction group was downregulated to (37±1)℃ using a semiconductor refrigeration instrument. The rats were transferred to the room with the temperature of 25℃ after the operation. Brain hypothermia was also induced in rats from the mild hypothermia group. At 13.0-14.0 minutes after establishing rat models of cerebral ischemia, the head on the side of cerebral ischemia was tightly connected with the probe of the semiconductor refrigeration instrument. The refrigerator temperature was adjusted to 6-8℃, so as to make the temperature of brain tissue on the lesion side at 32.0-33.0℃ for 4 hours.
RESULTS AND CONCLUSION:Compared with the cerebral infarction group, the BBB scores of rats inthe mild hypothermia group were distinctly increased, and the volume of infarcted area decreased. At 1 day after modeling, the expression level of growth associated protein 43 mRNA in brain tissue of rats in the mild hypothermia group was close to that in the cerebral infarction group. At 2 weeks after modeling, the expression level of growthassociated protein 43 mRNAin brain tissue of rats in the mild hypothermia group was significantly increased compared with that in the cerebral infarction group. These results suggest that mild hypothermia therapy can protect nerve cels against injury caused by cerebral infarction and promote the recovery of neurological function. Its underlying mechanism may be related to the up-regulation of the expression of growth associated protein 43 in ischemic penumbra .

AIM: PSP94 has been shown promise as a potential prostate cancer marker and it was reported that PSP94 can inhibit the growth of prostate cancer cell in vitro and in vivo. This study aimed to construct recombinant human PSP94 expression vector. METHODS:The PSP94 cDNA was obtained from normal prostate tissue, and recombinant plasmid pUC19-PSP94 was constructed. The target gene was identified and sequenced. Then the PSP94 gene was inserted to the secretory expression vector. RESULTS:The gene sequence of PSP94 was identified. The recombinant vector was constructed. The secreted PSP94 was isolated and identified by Western blot. CONCLUSION:The recombinated PSP94 could expressed PSP94 successfully.

This study is to investigate the binding capability of lidamycin apoprotein (LDP), an enediyne-associated apoprotein of the chromoprotein antitumor antibiotic family, to human breast cancer and normal tissues, the correlation of LDP binding capability to human breast cancer tissues and the expression of tumor therapeutic targets such as VEGF and HER2. In this study, the binding capability of LDP to human breast cancer tissues was detected with tissue microarray. The correlation study of LDP binding capability to human breast tumor tissues and relevant therapeutic targets was performed on breast cancer tissue microarrays. Immunocytochemical examination was used to detect the binding capability of LDP to human breast carcinoma MCF-7 cells. As a result, tissue microarray showed that LDP staining of 73.2% (30/41) of breast cancer tissues was positive, whereas that of 48.3% (15/31) of the adjacent normal breast specimens was positive. The difference between the tumor and normal samples was significant (Chi2 = 4.63, P < 0.05). LDP immunoreactivity in breast cancer correlated significantly with the overexpression of VEGF and HER2 (P < 0.001 and < 0.01, r = 0.389 and 0.287, respectively). Determined with confocal immunofluorescent analysis, LDP showed the binding capability to mammary carcinoma MCF-7 cells. It is demonstrated that LDP can bind to human breast cancer tissues and there is significant difference between the breast cancer tissues and the corresponding normal tissues. Notably, the binding reactivity shows positive correlation with the expression of VEGF and HER2 in breast carcinoma tissues. The results imply that LDP may have a potential use as targeting drug carrier in the research and development of new anticancer therapeutics. This study may provide reference for drug combination of LDM and other therapeutic agents.

To improve the inulinase application in biotechnology, the characteristic of inulinase from Kluyveromyces marxianus YX01 was investigated. The inu gene of K. marxianus YX01 was transformed into Pichiapastoris GS115 host cells with molecular biology techniques. Then we achieved the heterologous expression of exo-inulinase whose molecular mass was about 86.0 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Furthermore, six His-tag was added to the inulinase and a two-step method was applied in the purification of inulinase, including concentration via dialysis by polyethylene glycol 20 000 and metal Ni-NTA Agarose affinity adsorption. The purification factor of purified protein was 3.6 and the recovery rate of enzyme activity was 33.1%. We characterized the purified inulinase. The optimum temperature was 60 degrees C and pH was 4.62. When inulin and sucrose were used as substrates, the K(m) and V(max) values were 80.53 g/L vs 4.49 g/(L x min) and 183.10 g/L vs 20.20 g/(L x min), respectively. In addition, metal ions including Mn2+, Ca2+, Cu2+, Zn2+ and Fe2+ exhibited different degrees of inhibition on the enzyme activity, and Cu2+, Zn2+ and Fe2+ exhibited the most significant inhibition. Our findings might lay a good foundation for industrial application of inulinase.
Glycoside Hydrolases ; chemistry ; genetics ; Industrial Microbiology ; Inulin ; Kluyveromyces ; enzymology ; genetics ; Pichia ; Sucrose ; Temperature

Objective To investigate the effect of Akt on cardiomyocyte hypertrophy induced by hydrogen peroxide(H2O2).Methods The neonatal rat cardiomyocytes cultured in primary generation were treated with low concentrations of H2O2.The cardiomyocyte hypertrophy was evaluated by the determination of average cell volume and protein content.The effects of Akt inhibitor on cardiomyocyte hypertrophy induced by H2O2 was recorded.Western blot was performed to examine the phosphorylation of Akt induced by H2O2.ResultsH2O2 at 10 or 50 ?mol/L stimulated cardiomyocyte enlargement as measured by cell volume and the protein content per cell.The inhibitor of Akt inhibited the hypertrophic response of cardiomyoctes stimulated by H2O2.H2O2 increased the level of Akt phosphorylation in cardiomyocytes,while that is suppressed by the inhibitors of PI3K(phosphoinositide 3-kinase). Conclusion Akt signaling is involved in cardiomyocyte hypertrophy induced by H2O2.

This study is to optimize the preparation process of fusion protein Fv-LDP which was expressed in the form of inclusion body and consisted of lidamycin apoprotein LDP and single-chain Fv antibody (scFv) directed against type IV collagenase. The preparation and the dissolution of inclusion body, the immobilized metal affinity chromatography of the target protein and the renaturization by stepwise dialysis were optimized by single-factor analysis or orthogonal design. In addition, the refolded fusion protein Fv-LDP was refined by Sephadex G-75 chromatography followed by fluorescence-activated cell sorter (FACS)-based saturation binding assay to measure its antigen-binding activity. After optimization of the process, the purity of fusion protein Fv-LDP existed in the inclusion body was 63.9% and the corresponding solubility was 95.7%; Under denaturing conditions, the purity of fusion protein Fv-LDP was more than 95% after the purification process. The percentage of monomeric fusion protein Fv-LDP was 60% after the refolding process, while it was further refined to 85% which was 5.6-fold higher than that of the initial refolding condition. The refined fusion protein Fv-LDP could bind to human lung adenocarcinoma PAa cells and human hepatoma BEL-7402 cells with the dissociation constants (Kd) of 0.176 micromol x L(-1) and 0.904 micromol x L(-1), respectively. The preparation process of fusion protein Fv-LDP has been successfully optimized, which provides the experimental basis for the production and future development of fusion protein Fv-LDP, and might serve as a relatively practical system for the preparation of other scFv-based proteins expressed in the form of inclusion body.

OBJECTIVE: To evaluate the physical and mental health status of post-linguistic cochlear implantees, and then to explore the effectiveness on of different rehabilitation programs. METHOD: Mandarin hearing in noise test (MHINT), personal report of communication apprehension (PRCA-24) and Nijmegen cochlear implant questionnaire (NCIQ) were used to measure the hearing ability, mental health degree and the health related life quality in 36 post-linguistic cochlear implant users, respectively. The improvement of subjects' physical and mental health levels was compared with among different rehabilitation programs, including family training program, auditory habilitation program and psychological intervention program. RESULT: (1) Family training program only can improve the subject's hearing ability (P < 0.05), but failed to ease the communication apprehension; ((2) Auditory habilitation program can both significantly improve the subjects' hearing ability (P < 0.01) and ease the fear of talking face to face (P < 0.01); (3) Psychological intervention program can significantly increase the auditory abilities (P < 0.01), reduce the communication apprehension (P < 0.01) and improve the quality of life. CONCLUSION Post-linguistic cochlear implantees had obvious mental symptoms. It was very important to design an effective rehabilitation program to improvement the living quality of hearing loss people.
Anxiety ; Cochlear Implantation ; rehabilitation ; Cochlear Implants ; Deafness ; Hearing Loss ; Hearing Tests ; Humans ; Language ; Linguistics ; Mental Health ; Noise ; Quality of Life ; Speech ; Surveys and Questionnaires

Objective To study the inhibitory effect of hammerhead ribozyme targeting connective tissue growth factor(CTGF) on collagen I synthesis and cell cycle progression of human hepatic stellate cell line(LX-2) cells.Methods Hammerhead ribozyme cDNA targeting CTGF mRNA plus two self-cleaving sequences were inserted into pTriEx2 vector to construct a recombinant vector pTriCTGF-Rz.LX-2 cells were transfected with either pTriEx2 or pTriCTGF-Rz and further stimulated with or without TGF-1.There were five groups in the experiment:control group,pTriEx2 group,pTriCTGF-Rz group,pTriEx2 plus TGF-?1 group,and pTrCTGF-Rz plus TGF?1 group.Semi-quantitative RT-PCR was used to detect the levels of CTGF mRNA and collagen Ⅰ mRNA.ELISA and flow cytometry were used to detect the levels of collagen Ⅰ secretion and cell cycle.Results Transfection of pTriCTGF-Rz into LX-2 cells reduced the CTGF mRNA and collagen Ⅰ mRNA levels as well as collagen Ⅰ protein level compared with pTriEx2 group(P

Objective To preliminarily explore the effects and brain protective mechanism of intermittent hypoxia preconditioning ( IHP) on rats with seizures induced by lithium-pilocarpine ( Li-pilo) .Methods A total of 96 8-week old male Sprague Dawley rats ( clean grade ) were randomly divided into control group , seizure group and four IHP-seizure groups.The animal model of epilepsy was established by intraperitoneal injection of Li-pilo in the seizure group and four IHP-seizure groups (Li-pilo was injected at 1, 3, 7, or 14 days after a 5-day regimen of IHP).Subsequent seizure behavior , the latency period and percentage of generalized seizures were quantitatively evaluated for 240 min and the cognitive function was tested by Morris water maze task , and followed by the detection of hippocampus neuron apoptosis and related protein (BCL-2, Bax, and cleaved-caspase-3) by TUNEL labeling and Western blot, respectively.Results The induced seizure peaked on an average between 50-150 min after Li-pilo administration , scored using a modified Racine scale.The average scores of modified Racine scale in the IHP-3d seizure group was significantly lower than that in the other groups.The latency period and percentage of generalized seizures in the IHP-3d seizure group rats were significantly different from the parameters in the seizure group rats (P<0.05).IHP-3d seizure rats showed lower escape latency, neuronal apoptosis counts and higher percentage of time in the probe quadrant compare with the seizure group and the other three IHP-seizure groups ( P <0.05 ) .Compared with the control group , the parameters of water maze and apoptosis detection in the IHP-3d seizure group showed no significant changes (P>0.05).Conclusions The results indicate that IHP treatment may help to decrease the susceptibility to epilepsy by reducing abnormal apoptosis , and has a brain protective effect on the seizure rats .

Objective To explore the therapeutic effect of rhIL-11 and rhG-CSF on mouse bone marrow injury induced by neutron irradiation.Methods 130 male BALB/c mice were irradiated by 3.0 Gy neutron and mice peripheral blood cells,bone marrow pathological changes,bone marrow nucleated cell counts,AgNOR content,apoptosis and necrosis rates and Bax protein content were observed by means of blood cells automatic analyzer,HE staining,AgNOR staining,flow cytometry,immunohistochemistry staining and image analysis.Results In the irradiation group and the rhIL-11 group,the mice peripheral blood white blood cells,bone marrow nucleated cell counts and AgNOR content was decreased progressively.The Bax protein was positively or strongly positively expressed in the cytoplasm of the hematopoietic cells and the Bax protein content was increased progressively at 6 h,1 d,3 d after irradiation.In the irradiation group,the rates of apoptosis and necrosis in the mice hematopoietic cells were greatly increased and that of necrosis was significant at 6 h after irradiation.In the rhIL-11 + rhG-CSF group,the counts of bone marrow nucleated cell and AgNOR were increased and the Bax protein content was decreased at 3 d after irradiation,while in the rhIL-11 group,the indexes mentioned above were not obviously different compared with those of the irradiation group.Conclusions The mice bone marrow hematopoietic function is seriously damaged by 3.0 Gy neutron irradiation,rhIL-11 and rhG-CSF could improve the mice hernatopoietic function after neutron irradiation,and combination of them is more effective to stimulate the hematopoitic function than either of them alone.