Objective To observe the effect of pretreatment of aprotinin on nitric oxide (NO) and nitric oxide synthase (NOS) contents after ischemia-reperfusion injury of spinal cord in rabbits.Methods 45 rabbits were randomly divided into aprotinin treatment group (group A), normal saline control group (group B) and pseudo-surgical operation group (group C) with 15 rabbits in each group. The infrarenal segment in abdominal aorta was clamped for 60 min to construct the model of lumbosacral spinal cord ischemia in rabbits. Reperfusion was followed and kept on for 24 h until the blood flow regained normal. Aprotinin was given 3×107 IU/kg as a short time intravenous injection for 10 min before ischemia, and then was drilled with micro pump by 1×107 IU/kg/h. Normal saline was used in group B, the ischemia-reperfusion duration between group A and group B remained same. The group C was only exposured abdominal aorta and not clamped. The rabbits were killed before ischemia and at 8 h, 24 h after ischemia-reperfusion, lumbar segment spinal cords were harvested to detect contents of NO and NOS of spinal cord.Results After 8 h of ischemia-reperfusion,the contents of NO, total NOS (TNOS), and induced NOS (iNOS) in group A and group B were more than that before ischemia (P＜0.05). After 8 h of ischemia-reperfusion, there was a significant difference in the contents of NO, TNOS, iNOS between group A and group B (P＜0.05～0.01). After 24 h of ischemia-reperfusion, there was a significant difference too between group A and group B (P＜0.01). After 8 h and 24 h ischemia-reperfusion, the contents of NO, TNOS, iNOS in group A and group B were more than that in group C (P＜0.01).Conclusion During the ischemia-reperfusion, more NO produced is an important factor of spinal cord injury. Aprotinin can decrease the contents of NO and ischemia-reperfusion injury to spinal cord of rabbits.

Objective To observe the changes of MEP(motor evoked pontential) and neurologic function after Erigeron beviscapus(EBHM) preconditioning on spinal cord ischemia-reperfusion injury in rabbits, and provide substructural experimental theory.Methods Twenty-four domestic rabbits were randomly divided into four groups with six rabbits each. In sham group (group A) only the abdominal aorta was exposed. In the blank group (group B), the abdominal aorta was occluded 40 minute followed by reperfusion. In the methylprednisolone group (group C), it was occluded 30 minute before the abdominal, methylprednisolone (30 mg/kg) was injected into the rabbit by vein, which was used for two days after the surgeon. In the EBHM group (group D), EBHM(50 mg/kg) was injected into the rabbit by vein, the following procedure was the same as group C. MEP was measured in each observed point and neurologic function was observed everyday for 7days.Results At every observed point after SCI, the MEP changed regularly, and compared with group B, there was significant difference in groups C and D(P<0.05). 7 days later, the neurologic function changed significantly in each group(P<0.05).Conclusion MEP monitoring can reflect the early change on spinal cord ischemia-reperfusion injury. EBHM preconditioning can protect the rabbit from spinal cord ischemia reperfusion injury.

BACKGROUND:Insulin-like growth factor-1(IGF-1), a main active factor in growth hormone (GH), plays various biological functions, such as improving cognitive ability and anti-apoptotic action.
OBJECTIVE:To detect the expressions of GH and IGF-1 in the temporal cortex of Lewis dwarf rats, and to explore the effect of different concentrations of GH on the differentiation of hippocampal nerve stem cels (NSCs).
METHODS:Lewis dwarf rats aged 11(adult) and 20 (senile) month olds and normal wild-type rats were euthanized by decapitation, underwent the craniotomy quickly, and the temporal cortex in the cold saline was extracted. GH and IGF-1 levels were detected using western blotting. After isolation, purification and identification of the rat hippocampal NSCs, the effect of GH in different concentrations (10, 30, 90μg/L) on the NSCs differentiation was determined at 96 hours after culture.
RESULTS AND CONCLUSION:The GH level in the temporal cortex did not differ significantly among rats (P > 0.05). While the IGF-I level in the temporal cortex of Lewis dwarf rats was significantly higher than that of the wild-type rats (P < 0.05). The GH level in the temporal cortex of adult female Lewis dwarf rats was significantly lower than that of the male rats (P < 0.05). Immunofluorescence showed that the proportion ofβ III-tubulin-positive neurons was significantly higher than that in the control group (P < 0.05) after the hippocampal NSCs and precursor cels cultured for 96 hours with GH (30μg/L), but there was no significant difference between the control group and treatment group with GH of 10 or 90μg/L. These results suggest that GH and IGF-I are expressed in the temporal cortex of both Lewis dwarf and wild-type rats which are independent from pituitary GH and the peripheral circulating IGF-1. Additionaly, GH can promote the differentiation of hippocampal NSCs and precursor cels into neurons.

ObjectiveTo establish the rapid molecular diagnosis of 16 common coagulase negative Staphylococcus(CNS).MethodsDNA sequencing of 16 CNS would be obtained with gap gene.After the alignment gap gene sequences which were available in the GenBank,the bacteria were identified with homological alignment and phylogenetic tree,and compared with the 16S rRNA gene.ResultsThe sequence similarity of the gap sequences ranged from 39% to 98% in 16 CNS.There were the highest similarity (98%) between S.hominis and S.hominis subsp,and the lowest(39% ) between S.saprophyticus and S.xylosus.The sequence similarity of the 16S rRNA sequences ranged from 96 to 98%,at least two species of bacteria similar rate of 99% and the most four species similar rate of 99%.Phylogenetic homology analysis showed that it was a high confidence(99% ) in the detection ofS.xylosus and S.lentus,S.chromogenes and S.intermedius,S.hominis and S.hominis subsp,but for 10 other species of bacteria,gap homology analysis has less unreliable confidence(49%,56% ) and 16S rRNA has more unreliable confidence(43%,43%,50%,56%,63%,65%,76% ).ConclusionAnalysis of gap sequence could identify 16 CNS timely and accurately,with higher confidence than 16S rRNA.

ObjectiveTo evaluate the application value of the quantitative procalcitonin (PCT) test in bloodstream infection.Methods Of 1066 patients with blood culture and PCT detection were collected in our hospital,retrospectively,1010 were effective cases.The relationship between blood culture results and serum PCT levels was investigated.PCT levels in gram-negative bacterial infection,gram-positive bacterial infection and candidiasis were compared.The prognosis of 33 blood culture positive patients with repeated PCT detection results were analyzed.Mann-Whitney U test was used to compare the PCT value among the three groups,and Fisher' s test was used to compare the death rate among the three groups.ResultsIn the patients with negative blood culture results,the median of PCT was 0.37 (0.11 - 1.67) μg/L.But in the patients with positive blood culture results,the median of PCT were 2.24(0.57 -11.59) μg/L The positive rate of PCT in gram-negative bacteria infection,gram-positive bacterial infection and candidiasis were 86.6％,72.0％ and 75.7％,respectively.In the 33 patients subjected to repeated PCT detections,the mortality of the patients with decreasing PCT was lower than the others.The patients whose PCT levels were greater than 5 μg/L had poor prognosis.ConclusionsQuantitative PCT is proved to be an effective method for rapid diagnosis of bloodstream infection.The changing trends of PCT test results has certain reference value for the patients' prognosis.

Objective To investigate the effect of fatty acid synthase (FASN)on apoptosis in pancreatic cancer cell PANC-1 and the possible molecular mechanism.Methods Annexin V/FITC and flow cytometry were performed to detect the expression of FASN in pancreatic cancer PANC-1 after C75 treatment and the change of apoptosis in pancreatic cancer cell PANC-1 treated with C75.Quantity reverse transcriptase polymerase chain reaction (RT-PCR)and Western blot were used to measure the protein and RNA expressions of Caspase-3,bcl-2 and FASN.Results Inhibited by C75,the activity of FASN in pancreatic cancer cell PANC-1 was significantly decreased.Meanwhile,PANC-1 showed an increased apoptosis level in a dose-dependent manner (P < 0.05 ). Furthermore,after C75 inhibited FASN in pancreatic cancer cells,the protein and RNA expressions of Caspase-3 significantly increased (P <0.05)whereas the level of Bcl-2 reduced (P <0.05).Conclusion FASN is involved in the process of apoptosis in PANC-1 via Bcl-2 and Caspase-3.Therefore,FASN will provide a new target for the treatment of pancreatic cancer and generate better treatment efficacy.

Objective T o observe the chem ical groups changing in rat kidney w ith regard to fatal hyper-therm ia by Fourier transform infrared m icrospectroscopy (FT IR-M SP ) and to provide a new m ethod to diagnose fatal hypertherm ia. Methods R ats w ere sacrificed by hypertherm ia, brainstem injury, m assive hem orrhage and asphyxiation and divided into groups. T he renal sam ples w ere dissected im m ediately af-ter death. T he data of infrared spectroscopy in glom erulus w ere m easured by FT IR-M SP. Results T he absorbances of 3 290, 3 070, 2 850, 1 540 and 1 396 cm -1 significantly increased (P<0.05),and the ratios of A1650/A3290 and A1650/A1540 significantly decreased (P<0.05) in group of hypertherm ia. Conclusion FTIR-M SP can analyze the changes of chem ical groups of kidney as an auxiliary diagnosis for discrim inating hyper-therm ia w ith other causes of death.

Objective To screen and verify the proteins interacting with phosphorylation cluster of DNA dependent protein kinase catalytic subunit((DNA-PKcs) by yeast two-hybrid assay.Methods To know the proteins interacting with DNA-PKcs phosphorylation cluster,yeast two-hybrid assay was applied to screen the cDNA library of human hepatic tissue with a previously constructed plasmid pGBKT7-DPC.The positive clones were further identified by PCR,rotary validation and sequence analysis.Then the eukaryotic expression vectors of the bait protein and screened positive clone proteins were constructed and transfected into human embryonic kidney 293T cells to detect whether the proteins could been expressed correctly.At last,the bait protein and screened positive clone proteins were co-transfected into 293T cells and protein interaction was detected with Co-Immunoprecipitation (Co-IP) assay.Results After two rounds of screening using the yeast two-hybrid assay,12 candidate clones were obtained.Then 7 clones with different insert fragments were identified by PCR,and 3 positive proteins interacted with DNA-PKcs phosphorylation cluster were further verified by rotary validation.Sequencing analysis demonstrated that these 3 proteins were MBNL1,SIK2 and YY1AP1,respectively.Accordingly,the eukaryotic expression vectors of bait protein and 3 positive clone proteins were constructed successfully and expressed correctly in 293T ceils.Finally,the Co-IP assay confirmed that these 3 positive clone proteins could interact with DNA-PKcs phosphorylation cluster.Conclusions Proteins interacting with DNA-PKcs phosphorylation cluster are successfully screened and identified.

Objective To study the expression of tumor necrosis factor alpha (TNF-α) in the intestine of mice irradiated by neutron and γ rays.Methods 350 male BALB/c mice were irradiated with neutron and γ rays of different doses, and sacrificed at 6 and 12hours, 1, 2, 3, 4, 5, 7, 10, 14, 21 and 28 days after irradiation.The TNF-α in the mice intestinal tissue was detected by means of immunohistochemistry and image analysis.Results In normal control mice, TNF-α was expressed in the cytoplasm of macrophages in intestinal villus interstitium, submucosa and lymph tissue.After 2.5Gy neutron radiation, TNF-α was decreased progressively within 2 days, increased obviously in macrophages and crypt cells during 3rd～7th day, reached the peak at 5th day and recovered to normal level at 14th day and TNF-α was decreased progressively within 4 days after 4.0 and 5.5Gy neutron and 12Gy ray irradiation.TNF-α was increased obviously in 6～12 hours, decreased at 1st day, increased at 2nd～5th day, peaked at 3rd day and recovered at 10th day after 5.5Gy ray irradiation.Conclusion Neutron and ray radiation induce different expression profile of endogenous TNF-α in small intestine, which may be related with the pathologic courses of irradiation-induced damage and repair of intestine.

Sepsis-related deaths account for more than 50% of the total deaths in the world and are the primary problem threatening public health. Timely and rapid diagnosis is the key to improving the survival rate of patients with sepsis. Blood culture is a commonly used method to diagnose sepsis in clinics. However, it is time-consuming and unstable, which cannot used for pathogen quickly identification. In recent years, Microbiological rapid on-site evaluation (M-ROSE) based on morphological technique is applying to the clinics, showing significant advantages in timeliness and convenience, which would overcome the shortcomings of traditional diagnostic methods and be conducive to diagnosis of sepsis.