STUDY ON THE MECHANISM OF INTERFERON ACTION (Ⅺ)——EFFECT OF pppA2''''p5''''A2''''p5''''A ON THE LEVEL OF cAMP AND cGMP IN MACROPHAGES

In this paper, the increase of cellular cAMP and cGMP levels in macrophages induced bypppA2'p5'A2'p5'A (briefly 2'-5'P_3A_3) is first reported. The optimal concentration of 2'-5'P_3A_3 for the elevation of cellular cGMP to the highest level is 10~(-7)-10~(-6)mol/L, while thatfor cAMP is 10~(-7)mol/L. The time for cGMP to reach its peak value is 15 min and that forcAMP is 2 h, when the cells are treated with 2'-5' P_3A_3 at 10~(-7)mol/L, which is the optimalconcentration for developing biological effect of macrophages (phagocytosis). These resultssuggest that cGMP and cAMP may be related to, or may be the mediators for, 2'-5'P_3A_3action.     

MECHANISM OF INTERFERON ACTION——ACTIVATION OF MACROPHAGES AND ANTAGONISM TO a-FETO-PROTEIN BY _(ppp)A_(2'p5')A_(2'p5')A

_(ppp)A_(2'p),'A_(2'p,)'A(2'-5'P_3A_3) activates macrophages and increases the phagocytosis of macrophages from different species including human beings. This indicates that the activation of macrophages may be a general action of 2'-5'P_3A_3. This discovery broadens the effect of 2'-5'P_3A_3 beyond the antiviral field.α-Feto-protein (AFP) inhibits the phagocytosis of macrophages and may be involved in the development of hepatoma. Data presented here show that 2'-5'P_3A_3 can antagonize this suppressive effect of AFP.Methods used so far for introducing 2'-5'P_3A_3 into the cells were made with the aid of CaCl_2, etc. under conditions which may not be the same as those used clinically. It was found that 2'-5'P_3A_3 can develop its biological effect without the aid of CaCl_2, etc.     

MECHANISM OF INTERFERON ACTION——STRUCTURAL REQUIREMENT OF LIGAND AND CELLULAR SPECIFICITY

We had reported the existence of 2'-5'P_3A_3 receptor on the peritoneal macrophages from Wistar rat. In this paper data are given to demonstrate that the ligand, for binding with 2'-5'P_3A_3 receptor and enhancing the phagocytosis of macrophages, must have a three-phosphate radical on its 5'-terminus and at least three adenylate residues in 2'-5'oligoadenylate. (~3H)-2'-5'P_3A_3 can also bind on macrophages from other animals; and the binding of (~3H)-2'-5'P_3A_3 on Hepatoma cell line 7402 is lower than that on macrophages; whereas it is very low (binding ratio less than 1%) on erythrocytes (from rabbit, rat and mouse) or human embryo lung fibroblast cells. The binding conditions of the ligand with 2'-5'P_3A_3 receptor have also been investigated.     

MECHANISM OF INTERFERON ACTION——THE DISCOVERY OF pppA2'p5'A2'p5'A RECEPTOR

In this paper it is demonstrated that (~3H)2'-5'P_3A_3 can bind to the peritoneal macrophages from Wistar rat. This binding is strongly inhibited by cold 2'-5'P_3A_3 while the inhibiting capacity of 2'-5'P_3I_3 and 2'-5'A_3 is very small, showing that this binding is highly specific. And the binding of (~3H)2'-5'P_3A_3 is reversible, saturable and with high affinity. A Scatchard analysis of the binding data gives a linear plot, an apparent dissociation constant (Kd) about 1.3×10~(-7) M and binding sites per cell about 2.1×10~7. The above evidence shows the existence of the 2'-5'P_3A_3 receptor, a fact which, to our knowledge, has not been reported before.     

MECHANISM OF INTERFERON ACTION Ⅲ——SIGNIFICANCE OF _(ppp)A_(2'P5')A_(2'P5')A IN THE ANTIVIRAL ACTION OF INTERFERON

This paper is to evidence that the antiviral effect of _(ppp)A_(2'p5')A_(2'p5')A(2'-5'P_3A_3) could cover a wide spectrum of viruses, the RNA viruses such as influenza H_3N_2/77, influenza H_1N_1/77, ECHO_(11), rhino, Sendai, Sindbis and VSV, and the DNA viruses such as herpes (type Ⅰ). In addition, the antiviral effect of 2'-5'P_3A_3 on ECHO_(11) virus, as compared with other viruses, is more efficient than that of IFN itself. It seems likely that 2'5'P_3A_3 plays an important role in the antiviral action of IFN. After comparing the action of 2'-5'P_3A_3 with interferon (IFN), it has appeared that the action of 2'-5'P_3A_3 is something parallel to that of IFN for most viruses. For a few viruses, however, considerable differences have been observed.     

SOLID-PHASE SYNTHESIS OF 2''''—5'''' OLIGOADENYLATE——2''''—5''''A CORE

With phosphotriester approach, the 2'—5' linked oligonucleotide——2'—5'A core was synthesized by using cross-linked polyacrylmorpholide resin as the support and the mesitylene sulfonyl chloride and N-methylimidazole as the condensing agent. Synthesizing the 2'—5'A core with this method has not yet been described. The yields that each of the protected monomers was attached to the polymeric support were satisfactory, i.e., 100%, 60% and 91% respectively.For the synthesis, we improved the methods for preparing the polymeric support and a series of intermediates. For this reason, the yields of the products were markedly increased and the operations to separate the products from the by-products and reagents were simplified.     

PURIFICATION AND KINETICS OF MOUSE LIVER FRUCTOSE 6-PHOSPHATE, 2-KINASE

The mouse liver fructose 6-phosphate, 2-kinase was purified by ultracentrifugation, polyethylene glycol precipitation, and subsequently by chromatography on DEAE-Sephadex, Blue-Sepharose and phasphocellulose columnS. Gel filtration and SDS polyacrylamide electrophorcsis showed that the enzyme has a molecular weight of 110,000 with two identical subunits. Mg~(2+) is essential for its activity. The activation of the enzyme by Mg~(2+) showed a positive cooperativity. The substrate saturation curve for fructose 6-phosphate was sigmoidal and for ATP was hyperbolic. The K_m's for ATP increased with decrease in concentrations of fructose 6-phosphate indicating that the sequence for the substrates binding was in an ordered mechanism with respect to fructose 6-phosphate prior to ATP. An ionizable residue at the active site with pKa 9.5 was essential for the ATP binding and the pKa shifted to 9.8 after the binding of ATP.     

干扰素作用机理的研究——Ⅺ.2′-5′三腺苷酸对巨噬细胞中cAMP和cGMP含量的影响

本研究首次证实了干扰素的作用介质pppA 2'p5'A2'p5'A(2'-5'P_3A_3)能引起巨噬细胞中cGMP,cAMP含量的变化。发现不同浓度的2'-5'P_3A_3和同一浓度的不同作用时间,对细胞内两种环核苦酸的含量有不同的影响。2'-5'P3_A_3的最终浓度在1×10~(-7)—1×10~(-6)mol/L时,可以引起细胞内cGMP显著升高,而使cAMP达到高峰的2'-5'P_3A_3浓度则为1×10~(-7)mol/L。细胞中cGMP与cAMP出现高峰的时间也显著不同,cGMP出现高峰的时间是在加入2'-5'P3_A_3以后15min,而cAMP出现高峰时间则在保温2h之后。上述结果提示,cGMP与cAMP可能与2'-5'P_3A_3受体发挥作用有某种关系。     

Computational Investigation on the Allosteric Modulation of Androgen Receptor

Androgens have similar structures with different biological activities. To identify molecular determinants responsible for the activity difference, we have docked six steroidal androgens to the binding site or the surface of androgen receptor by using molecular docking with computational investigation. The energy was calculated respectively based on the QM (quantum mechanics) and MM (molecular mechanics) methods. The result shows that the allosteric modulation of androgen receptor plays an important role in the binding process between androgens and receptor. The open state receptor is less stable than the close state one, but the latter is more favorable for binding with androgens. It is worthy of note that when the androgen receptors binding or without binding with androgen are in close state, they are difficult to return to their open state. This phenomenon is an exception of the well known two-state model theory in which the two states are reversible. Whether the internal of close state androgen receptor has a combination of androgen or not, the androgen receptor surface can be combined with another androgen, and their surface binding energies could be very close. The result is consistent with the experimental observations, but this phenomenon of continuous combination from open state is also an exception of the two-state model theory.     

LABELING OF PROTEIN WITH A NEW EUROPIUM CHELATE OF 5-CHLOROSULFOYL-2-THENOYLTRIFLUOROACETONE(CTTA)AND APPLICATION TO IMMUNOASSAY

We describe for the first time the synthesis and the optimal conditions for proteinlabeling with a new fluorescent probe,5-chlorosulfoyl-2-thenoyltrifluoroacetone(CTTA),whicb forms ahighly fluorescent conplex with Eu~(3+) when conjugated to protein.The labeled proteins werecharacterized by absorbance and fluorescence measurements and the effect of labeling on thebiological activity of sone proteins was also studied.It is shown that the new label is suitablefor applications in time-resolved fluoroimmunoassay.     

IONIC CONDUCTIVITY AND CATALYTIC PERFORMANCE OF A_2BO_4/La_2O_3 CATALYSTS FOR OXIDATIVE COUPLING OF METHANE

A series of La_2O_3 cartalysts modified by A_2BO_4 compounds with supe-rionic conductivity were prepared. It was observed that the addition of A_2BO_4compounds to La_2O_3 enhances C_2 yields, and the C_2 yields over the modified cat-alysts are intimately related with their conductivities. The effects of A_2BO_4compounds on the conductivity and catalytic performance were discussed. Onlywhen ion A has high conductivity, can the catalyst show good catalytic perfor-mance. A mechanism for interpreting the significant effects of alkali metals wasproposed.     

EFFECT OF CHRONIC IRRADIATION OF ~(60)Co-γ-RAY AT LOW DOSE RATE ON RHESUS MONKEY——EFFECT ON PERIPHERAL BLOOD AND TESTIS

The authors have demonstrated that after a long-term chronic irradiation of 2.17 rad/day for 2 months, the testis of the experimental monkey (Macaca mulatta) is extremelyatrophied with a loss of reproductive ability. But the effect on peripheral blood is notapparent even after irradiation for 57 months. It shou1d be noted that rhesus monkeys, being as sensitive to irradiation and similar inevolutionary steps as human beings, could be used as experimental animals to further studythe dose rate for aspermatogenesis, the morphological injuring effect on the reproductivesystem and the cytogenetic effect of irradiation, and this is highly important to working outa standard of promised dose of irradiation for X-ray workers and an index of radiationprotection monitoring.     

OHMEFENTANYL——A NEW AGONIST FOR μ-OPIATE RECEPTOR

Ohmefentanyl (F 7302, N[1-(β-hydroxy-β-phenylethyt)-3-methyl-4-piperidyl]-N-phenylpropionamide) is a potent synthetic analgesic agent. The analgesic activity of ohmefentanyl in mice is 6300 times more potent than that of morphine. The potency of ohmefentanyl in competing with specific binding of ~3H-naloxone is reduced by Na~ (100 mM) and GTP(50μM), thus suggesting the agonist properties of this compound. Binding characteristics of ~3Hohmefentanyl with mice brain P_2 fraction are studied. An important saturable, specific and reversible binding is demonstrated. Scatchard analysis indicates the existence of two classes of binding sites (KD_1=0.32nM, KD_2=3.91nM). Various opiate drugs strongly inhibit the binding of ~3H-ohmefentanyl, but nonopiate drugs have negligible affinity. Comparison of the relative potencies of morphine, DSTLE(Tyr-D-Ser-Gly-Phe-Leu-Thr, a specific ligand for the δ-opiate receptor) and ohmefentanyl in competing with ~3H-dihydromorphine (μ) and ~3H[D-Ala~2, D-Leu~5]-enkephalin (     

EFFECT OF PROCESSING METHOD ON THE IMPACT STRENGTH OF POM/TPU/CaCO3 TERNARY COMPOSITES

Polyoxymethylene （POM）/elastomer/filler ternary composites were prepared, in which thermoplastic polyurethane （TPU） and inorganic filler, namely, CaCO3, were used to achieve balanced mechanical properties of POM. The dispersion and phase morphology of POM/elastomer/filler composites were found to depend largely on processing method, CaCO3 content in masterbatch and the filler size. Two processing methods were employed to prepare POM/elastomer/filler ternary composites. One is called the one-step method, in which elastomer and the filler directly melt blended with POM matrix. The other is called the two-step method, in which the elastomer and the filler were mixed to get masterbatch first, which was then melt blended with pure POM of different content. The effect of phase morphology and processing method on impact strength was investigated. It was found that the two-step method results in an increase in impact strength but not for the one-step method. Additionally, the impact strength of POM ternary composites decreases with the increase in the size of CaCO3 particles.     

STRUCTURAL RULES OF TRANSITIONMETAL CARBONYL COMPOUNDS

In this article we have proposed two topological rules to account for the electronic structures of transition-metal carbonyl compounds. The second rule is applied to those which have polyhedral metalhc skeletons with triangular faces and their derivates, while the first rule is applied to all the others We have used these two rules to analyze 261 structure-known compounds with the number of metal atoms from 2 to 12, and the results show that the two rules are in good agreement with the experimental facts. Furthermore, we have also derived from the second rule a formula which can be used to account for the electronic structures of closedpacking carbonyl compounds and which is the same as that given by Ciani and his coworkers.     

THE STRUCTURE BASIS OF THE POOR FIBRIN SPECIFICITY OF UROKINASE(Ⅱ)——THE INHIBITION OF UROKINASE A CHAIN 149--157 ON THE FIBRIN STIMULATED ACTIVATION OF PLASMINOGEN BY TISSUE TYPE PLASMINOGEN ACTIVATOR

In view of the similarity of the charge distribution between fibrin A_α148--161 and Achain 149--157 of urokinase,the latter might compete with fibrin A_α148--161 when singlechain pro-urokinase is converted to double chain urokinase.To test this, the stretch of uro-kinase A chain 135--157 was separated from the low molecular weight urokinase, a competi-tive binding between this stretch and fibrin to tPA kringle-2 was shown by radio-bindingassay. The inhibition of the stretch on the fibrin stimulated activation of plasminogen wasdemonstrated in the caseinolytic system. The synthesized novapeptide urokinase A chain 149--157 (R-peptide) showed a significant inhibition on the activation of plasminogen in the pres-ence of fibrin. By contrasting finely with R-peptide, a synthesized novapeptide in which Arg154and Arg156 were replaced by Asp (D-peptide) did not show any inhibition effect on the fi-brin stimulated activation of plasminogen by tPA. These results suggest that the positivelycharged residues in the     

EFFECTS OF MONOCLONAL ANTIBODY ANTIEGF RECEPTOR ON HUMAN NASOPHARYNGEAL CARCINOMA CELL AND OTHER TUMOR CELLS

Three anti-EGF receptor MoAbs were used in these studies. Administration of MoAbs 3 and 176 inhibited tumor formation in nude mice by CNE-2, a poorly differentiated nasopharyngeal carcinoma cell line and A431, an epidermoid carcinoma cell line. When the same MoAbs were used in treatment against HeLa, a cervical carcinoma, tumor growth was not affected. The number of EGF receptors and apparent dissociation constants for ~(125)I-EGF on CNE-2 and A431 was 1.3×10~(?)/cell (Kd 7.7×10~(-8)mol/L) and 1.4×10~6/cell(Kd 2.4×10~(-9)mol/L), respectively. Both MoAbs 3 and 176, capable of competing with EGF for receptor binding, showed significant tumor growth inhibition. MoAb 101 was incapable of blocking the binding of EGF to its receptor, and not as effective as MoAbs 3 and 176 in tumor growth inhibition. Our observation is that the MoAb anti-EGF receptor is cytostatic rather than cytocidal, in vitro against CNE-2 and A431.     

ROLE OF THIOL-DISULFIDE EXCHANGE IN INSULIN BINDING TO ITS RECEPTOR

The effects of reduction by DTT, oxidation by DTNB and treatment with NEM on the thiol contents and insulin binding to its receptor in mice liver membranes were studied. Reduction with DTT leads to a parallel increase in the thiol content and the speciflc binding of insulin to the membrane. Scatchard analysis of the results shows little change in the number of binding sites but a twofold increase of the binding constant. Washing the membrane with bound insulin by a DTT containing buffer results in a more marked increase in the release of bound insulin than washing with buffer alone, suggesting that part of the insulin is bound to its receptor by covalent disulfide linkages through a thIol-disulfide exchange reaction and reduction with DTT leads to a marked increase in this "disulfide-linked" insulin. Treatment with DTNB or NEM of the DTT-reduced membrane seems to reverse the effect of DTT reduction, although the reaction of the untreated membrane with DTNB or NEM had little or no effect on the specific     

[~3H] TYROSINE BINDING TO PLASMA MEMBRANE PREPARATION OF RAT CORPORA LUTEA

Plasma membrane preparations of rat corpora lutea have been incubated with [~3H]tyrosine. [~3H]-tyrosine binding sites are demonstrated and Scatchard analysis shows that there exist two types of binding sites, one with high affinity and low capacity, the other with low affinity and high capacity. The kinetics studies demonstrate that the [~3H]tyrosine binding to the two types of binding sites is reversible and the speed of binding to the high affinity type is faster than that to the low affinity type. The analysis of the chemical structure of tyrosine analogues and related compounds with respect to the specificity of the binding sites reveal that both types of binding sites show specificity, but the specificity of the high affinity sites is higher than that of the low affinity sites. The relations of tyrosine structure to binding processing and to tyrosine inhibitory action on hCG-induced progesterone production are discussed. It is suggested that the high affinity binding sites might be regarded as "ty     

CRYSTAL AND MOLECULAR STRUCTURES OF TWO ISOMERS OF DIETHYL 2, 3-DICYANO-2, 3-DIPHENYLSUCCINATE

Diethyl 2, 3-dicyano-2, 3-diphenylsuccinate was prepared through the oxidative coupling of ethyl α-cyanophenylacetate with Cu~(++)-TMEDA-O_2 system. The product gave, upon chromatography, two crystalline compounds 1 and 2. The single crystals of 1 and 2 were identified by X-ray crystallographic analysis to be meso-and racemie-diethyl 2,3-dicyano-2,3-diphenylsuccinate respectively. It is noteworthy that in the molecules of 1 and 2 there is a remarkable lengthening effect of the bond length for the central C—C bond, i. e. 1: 1.585; 2: 1.62. It is also observed that there is steric hindrance between the substituent groups attached to the two carbon atoms of the central C—C bond respectively. These data provide the structural basis for the facile dissociation into radicals and the action of polymerization initiator of the title compound. CNDO/2 calculation of charge density and Wiberg bond order for compound 1 yields the results, which are consistent with the crystallographic data.