1,6二磷酸果糖在心肺复苏中的作用

１，６二磷酸果糖在心肺复苏中的作用无锡市第五人民医院急诊科张仲俭大剂量肾上腺素的使用提高了心肺复苏（ＣＰＲ）中自主循环的成功率［１，２］，从而也观察到心脏复跳后的许多异常心电图改变。本文分析我科９例心脏复跳后使用１，６二磷酸果糖（ＦＤＰ）对心血管的明...

高纯度1，6一二磷酸果糖分离精制新工艺

本文报道了１，６一二磷酸果糖（ＦＤＰ）发酵液通过沉淀一吸附一重结晶法，代替国外反渗析法或联合柱色谱分离一分子膜过滤一冷冻干燥法分离精制ＦＤＰ的新工艺。本工艺简便地解决了反渗析法纯化过程中高达５０％ＦＤＰ严重损失问题，避免了过程冗长的柱色谱分离过程易引起的细菌性污染所导致的ＦＤＰ破坏和复杂价昂的分子膜过滤器及冷冻干燥，突破了ＦＤＰ难以结晶的关键技术。所用设备简单，便于工业生产。经４升规模试验研究表明，ＦＬＰ总收率达８０％，纯度达９９％，高于国外文献报道及意大利进口冻干粉指标。

Structure and Expression Analyses of a Gene Encoding Fructose-6-Phosphate, 2-Kinase/Fructose-2,6-Bisphosphatase from Maize

A full-length cDNA encoding fructose-6-phosphate, 2-kinase/fructose-2,6-bisphosphatase from maize (Zea mays L.) was cloned by the methods of reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE), and designated as mF2KP. The encoded protein is composed of two regions. Its COOH-terminal region is catalytic region and homologous to the enzymes from other eukaryotes; and its NH 2-terminal region is common and special region only in plant. A truncated fragment of mF2KP covering integrated catalytic region was expressed in Escherichia coli. The fusion protein had the activities of fructose-6-phosphate, 2-kinase as well as fructose-2,6-bisphosphatase. Northern blot showed that the transcript level of mF2KP in seedlings initiated from strong-vigor seeds is lower than that from weak-vigor seeds.

Protective Effects of Sodium Magnesium Fructose Diphosphate on Brain Damage of Rats Subjected to Focal Cerebral Ischemia and Reperfusion

Objective: To study the effects of sodium magnesiusm fructose diphosphate(FDPM) on brain damage of rats after ischemia-reperfusion. Methods: Rats were subjected to cerebral ischemia-reperfusion induced by inserting a nylon thread into internal carotid artery to block the origin of middle cerebral artery and removing the thread later. FDPM (400 mg·kg -1), fructose-1,6-diphosphate (FDP, 400 mg·kg -1)and magnesium sulfate (MgSO 4, 30 mg·kg -1) were administrated 10 min after the onset of ischemia. Neurological scale, brain infarct area, Malondialdehyde(MDA) content and histopathological changes of brain tissue were studied. Results: FDPM decreased neurological scale, diminished brain infarct area, reduced MDA content and relieved histopathological change of rat brain tissue subjected to ischemia-reperfusion. These effects were more powerful than that of FDP or MgSO 4. Conclusions: It is suggested that FDPM markedly prevented rats against brain damage after cerebral ischemia-reperfusion, and its effect was better than that of FDP or MgSO 4.

Protective Effects of Oral Fructose-1, 6-diphosphate on Liver Injury in Animal Models

Aim To investigate the effects of FDP on different liver injury models to explore the possibility of FDP used as an oral liver protective agent. Methods Chronic liver injury model in rats was induced by carbon tetrachloride （ CCl4 ） ; Acute liver injury model in mice was induced by aminogalactose （GAIN） or lipopolysaccharide （LPS）. Results In CCl4-induced chronic liver injury model, FDP （1 , 4 g·kg^-1·d^-1, q.d., for 10 weeks） significantly lowered ALT, AST,γ-glutamyl transpeptidase （γ-GT）, alkaline phosphatase （ALP）, and total bilirubin （T-BIL） in serum compared with vehicle; simultaneously it evidently elevated abnormal total protein （TP）, albumin （ALB） and total cholesterol （ T-CHO ） levels in serum; it also dose-dependently reduced hydroxyproline contents in hepatic tissue. 4 g·kg^-1·d^-1 of FDP apparently decreased incidence of hepatic cirrhosis, and alleviated pathological changes of liver tissue. In GaiN-induced model, 1.0 - 4. 0 g·kg^-1·d^-1 of FDP （ bid, for 3 d ） significantly lowered alanine aminotransferase （ ALT ） and aspartate aminotransferase （ AST ） levels in serum ; it also decreased liver coefficient. 4. 0 g·kg^-1·d^-1 of FDP significantly alleviated pathological changes of cell ultra-structures. In LPS-induced model, only high dose of FDP （4. 0 g·kg^-1·d^-1, bid, for 12 d） significantly decreased ALT level in serum. Conclusion This study first demonstrated the protective effect of oral FDP on chronic liver injury caused by CCl4, and confirmed its effect on acute liver injury at the same time, suggesting that Long-term oral FDP is efficacious against liver injury induced by different factors and can be used as an oral liver protective agent in clinic.

Deletion of 93 bp Far-upstream Fragment of Rice Cytosolic Fructose- 1, 6-Bisphosphatase Promoter Completely Alter Its Expression Pattern

The 1 195 bp 5′ flanking region of rice ( Oryza sativa L.) cytosolic fructose_1, 6_bisphosphatase (cyFBPase) can direct tissue, cell specific expression in transgenic rice. In order to identify sequence elements responsible for the regulation of mesophyll_specific expression, the 5′ flanking regions of -1 195 bp, -1 102 bp, -768 bp, and -644 bp upstream of the translation initiation ATG codon were fused to the reporter gene encoding β_glucuronidase (GUS) and transferred to rice via particle bombardment. Analysis of the 5′ promoter deletions identified that a 93 bp fragment between -1 195 bp and -1 102 bp is essential for directing mesophyll specific expression. High constitutive expression of GUS reporter gene was found in the -768 deletion lines and another two deletion series. These results indicate the great potential utility of the promoter in rice biotechnology.

Isolation of a 1 195 bp 5′-Flanking Region of Rice Cytosolic Fructose-1,6-bisphosphatase and Analysis of Its Expression in Transgenic Rice

A genomic DNA fragment containing the 5'-upstream sequence and part of the open reading frame corresponding to the cytosolic fructose-1,6-bisphosphatase (cyFBPase) cDNA was isolated by Genome Walking. The 1 195 lip 5'-flanking region which started from the translation initiation ATG codon was fused to reporter gene encoding beta-glucuronidase (GUS) and stably transferred to rice via particle bombardment. Strong GUS activity was detected in leaves and leaf sheaths of transgenic rice, but not in culms and roots. Histochemical localization revealed that GUS expression was exclusively restricted to mesophyll cells in transgenic rice. Our results indicate that the 1 195 bp fragment contains all the cis-elements required for directing mesophyll-specific expression pattern in rice.

Protective Effect of Tetrandrine and Fructose-1,6-diphos phate on the Model of Focal Cerebral Ischemia in Rats

The effect of tetrandrine (Tet) on the infarction area and volume of rat brain induced by middle cerebral artery occlusion (MCAO) was investigated. The treatment with Tet 7.5, 12.0 or 15.0 mg·kg 1 , or with fructose 1,6 diphosphate (FDP) 200 and 350 mg·kg 1 ip immediately after MCAO, respectively, significantly reduced the infarction area and volume in a dose dependent manner. MK801 and FDP also displayed a protective effect on brain ischemia. A combination of Tet and FDP administered immediately after MCAO, produced a more potent protective effect than those treated with Tet or FDP alone. When Tet or FDP was administered 1 h and 2 h after MCAO, respectively, they could still significantly reduce the infarction area and volume of brain tissue. But, there was no significant protective effect when these two compounds were given 3 h after MCAO.

Effects of fructose-1,6-diphosphate on concentration of calcium and activities of sarcoplosnic Ca^(2+)-ATPase in cardiomyocytes of Adriamycin-treated rats

Objective: To observe the effects of fructose-1,6-diphosphate (FDP) on serum levels of cardiac troponin 1 (cTnl) and creatine kinase-MB (CK-MB), as well as the concentration of calcium in cardiomyocytes (Myo[Ca2+]) and activity of sarcoplosnic Ca2+-ATPase (SRCa2+-ATPase) in Adriamycin (ADR)-treated rats. Methods: Rats were intraperitoneally injected with ADR (2.5 mg/kg every other day for 6 times) and then with different dosages of FDP (every other day for twenty-one times). Bi-antibodies sandwich Enzyme linked immune absorption assay (ELISA) was performed to detect serum level of cTnl. CK-MB was detected by monoclonal antibody, Myo[Ca2+] was detected by fluorescent spectrophotometry and the activity of SRCa2+-ATPase was detected by inorganic phosphate method. Results: FDP (300, 600, 1200 mg/kg) significantly reduced the serum levels of cTnl and CK-MB, while at the same time decreased calcium concentration and increased SRCa2+-ATPase activity in cardiomyocytes of ADR-treated rats (P<0.01). Conclusions: FDP might alleviate the cardiotoxic effects induced by ADR through decreasing calcium level as well as increasing SRCa2+-ATPase activity in cardiomyocytes.

A New Method of Crystallization of Octahydro Trisodium Salt of Fructose- 1,6-diphosphate

In order to overcome the elementary heterogeneous nucleation whileoctahydro trisodium salt of fructose- 1, 6-diphosphate (FDPNa_3·8H_2O) is crystallized with ethanol precipitation at low temperature,a new crystallization method with alcohol precipitation combined withsalt precipitation has been presented. The ethanol-sodium ac- etatesystem for crystallization of salt of fructose-1, 6-diphosphate isbased on the mechanism of crystallization of FDPNA_3·8H_2O in theethanol-low temperature system. It is found that crystal size may becontrolled by regulating Temperature of pH value of solution in thecrystallization process, and the crystal yield increases to 95/100from 78/100 Which obtained in the ethanol-low temperature system.

Fructose-1,6-diphosphate-added total parenteral nutrition in septic animals and stressed patients

Objective To investigate the roles of fructose-1,6-diphosphate(FDP)-added total parenteral nutrition (TPN)in septic animals and stressed patients.Methods Thirteen adult dogs were randomly assigned to one of two study groups 6 hours after the induction of severe intra-abdominal infection.Group TPN(n =6)received 70 kcal· kg-1· d-1 of nonprotein calorie(NPC)and 0.56 g· kg-1· d-1 of nitrogen.1 g/kg of FDP was also infused to the animals in group TPN + FDP(n = 7)everyday.In the clinical study,the control group received routine TPN,while the study group(n = 16)was treated with TPN plus FDP(5 g,two times a day)for 7 days.Results In dogs with TPN support,plasma ATP levels were not changed significantly,while the value in the TPN + FDP group increased significantly from 0.18 μmol/L to 0.46 μmol/L at 24 h and 0.51 μmol/L at 48 h(P ＜ 0.01).Muscular ATP increased markedly in the TPN + FDP group.Muscular creatine phosphate alues were not significantly changed in the TPN group,but the values increased in the TPN + FDP group from 4.06 μmol/g·wt at the beginning to 4.93 μmol/g· wt at 24 h and 5.60 μmol/g·wt at 48 h(P ＜ 0.05),with a cytochrome oxidase increase in immunohistochemistry stain.In the clinical study,plasma ATP levels increased and urinary 3-methylhistidine production significantly decreased with an improved value for positive accumulative nitrogen balance in the FDP-infused group.Conclusion Our results suggest that total parenteral nutrition support with the supplement of fructose-1,6-diphosphate has a positive role in body energy production and protein metabolism in septic animals and stressed patients.

Beneficial effects of fructose 1,6 diphosphate on hemorrhagic shock in rats

Objective: To investigate the effects of fructose 1,6 diphosphate (FDP) on experimental hemorrhagic shock in rats. Methods: Sixty rats were randomly divided into three groups: the normal saline control group (group A), the 5% glucose solution control group (group B) and the 5% FDP solution treated group (group C). Shock models were made by bloodletting until the mean arterial pressure (MAP) reduced to 39.75 mmHg (1 mmHg=0.133 kPa) for 60 minutes, and then normal saline, 5% glucose and FDP were given to the rats, respectively. Results: FDP could significantly increase MAP and the survival rate, elevate pH value, partial oxygen pressure (PaO 2) and superoxide dismutase (SOD) activity, and decrease partial carbon dioxide pressure (PaCO 2) and malondialdehyde (MDA) in arterial blood of the shocked animals. Conclusions: It suggests that FDP has a good protective effect on hemorrhagic shock by improving tissue metabolism and preventing acidosis and tissue injury caused by free radicals.