[Clinical, biochemical and gene mutation characteristics of short chain acyl-coenzyme A dehydrogenase deficiency by neonatal screening].

Objective: To investigate the incidence, clinical, biochemical and gene mutation characteristics of short chain acyl-coenzyme A dehydrogenase deficiency (SCADD). Method: From January, 2009 to October, 2015, a retrospective analysis of the urine organic acids and acyl-coenzyme A dehydrogenase (ACADS) gene mutation characteristics of patients diagnosed as SCADD by newborn screening using tandem mass spectrometry in Department of Genetics and Metabolism (Newborn screening Center of Zhejiang Province), Children's Hospital, Zhejiang University School of Medicine. Dietary guidance, life management and supplementation of L-carnitine were conducted, and growth and intelligence development were observed during follow-up among the SCADD patients. Result: A total of 1 430 024 neonates, seventeen cases were diagnosed with SCADD with an incidence of 1/84 117. All patients had no clinical symptoms, and intelligence and physical development were normal. Blood butylacyl-carnitine (C4) levels and the ratios increased, C4 0.713.14 µmol/L(reference value 0.03-0.48 µmol/L), C4/C2 0.07-0.23(reference value 0.01-0.04), C4/C3 0.65-2.04(reference value 0.05-0.39). Thirteen with increased urinary ethyl malonic acid (9.30-90.99 mg/g creatinine (reference value 0-6.20 mg/g creatinine )), one patient was accompanied by increased methyl succinic acid (12.33 mg/g creatinine(reference value 0-6.40 mg/g creatinine)), one subject with increased acetylglycine (3.52 mg/g creatinine(reference value 0-0.70 mg/g creatinine)). A total of 13 known mutations were detected in the ACADS gene, 1 homozygous mutation (c.1031A>G), the others are compound heterozygous mutations. One frameshift mutation (c.508_509delGC) and 12 missense mutations were detected. Common mutation were c. 1031A>G(35.3%), c. 164C>T(20.6%) and c. 991G>A(11.8%). SCADD in newborn screening program had no clinical symptoms and normal growth development after 8-42 months follow-up. Conclusion: Cases with SCADD had no clinical symptoms with an incidence of 1/84117. The c. 164C>T and c. 1031A>G may be the common mutations.

Effects of maternal folic acid supplementation on gene methylation and being small for gestational age.

BACKGROUND: Being small for gestational age (SGA), a foetal growth abnormality, has a long-lasting impact on childhood health. Its aetiology and underlying mechanisms are not well understood. Underlying epigenetic changes of imprinted genes have emerged as a potential pathological pathway because they may be associated with growth, including SGA. As a common methyl donor, folic acid (FA) is essential for DNA methylation, synthesis and repair, and FA supplementation is widely recommended for women planning pregnancy. The present study aimed to investigate the inter-relationships among methylation levels of two imprinted genes [H19 differentially methylated regions (DMRs) and MEST DMRs], maternal FA supplementation and SGA. METHODS: We conducted a case-control study. Umbilical cord blood was taken from 39 SGA infants and 49 controls whose birth weights are appropriate for gestational age (AGA). DNA methylation levels of H19 and MEST DMRs were determined by an analysis of mass array quantitative methylation. RESULTS: Statistically significantly higher methylation levels were observed at sites 7.8, 9 and 17.18 of H19 (P = 0.030, 0.016 and 0.050, respectively) in the SGA infants compared to the AGA group. In addition, the association was stronger in male births where the mothers took FA around conception at six H19 sites (P = 0.004, 0.005, 0.048, 0.002, 0.021 and 0.005, respectively). CONCLUSIONS: Methylation levels at H19 DMRs were higher in SGA infants compared to AGA controls. It appears that the association may be influenced by maternal peri-conception FA supplementation and also be sex-specific.

Structural and magnetic properties of ZnO nanocrystals in (Zn, Al)O films using pulse laser deposition.

In this manuscript, we reported that the room temperature ferromagnetism was observed in (Zn0.70, Al0.30)O film, which was fabricated by a novel physical method (pulse laser deposition (PLD)). The film was deposited from (Zn0.80, Al0.20)O ceramic target onto quartz (110) substrate by PLD at 400 degrees C under an oxygen partial pressure of 10(-4) torr. TEM result shows ZnO NCs with diameter of 4-5 nm and they are quite uniformly embedded into amorphous ZnO-Al2O3 phase. The SAED shows clearly that ZnO NCs possess polycrystalline structure. The SQUID measurement shows that the film has room temperature ferromagnetism (saturation magnetization = 3.6 emu/cm3) with Curie temperature above 300 K. The magnitude of magnetic moment of the films can be changed by tuning ZnO NCs size. Both oxygen partial pressure and film thickness studies show that the origin of ferromagnetism is possibly related to the oxygen defects at the surface of ZnO NCs.

[Effect of rhubarb on intestinal immune associated secretion in healthy mice and in burn mice].

OBJECTIVE: To explore the therapeutic mechanism of rhubarb (sennosides) in protecting intestinal muco-membranous barrier by observing the change of intestinal immune associated secretion (IIAS) in mice before and after burn and the enhancing effect of rhubarb on it in healthy and in burn mice. METHODS: Bal b/c mice were divided into 4 groups. Group A (n = 8), untreated healthy mice; Group B (n = 11), healthy mice treated with 10% rhubarb decoction 12 ml/kg, once every 6 hrs; Group C (n = 7), the mice with 30% back full-thickness burn and no treatment was given; Group D (n = 9), the mice with 30% back full-thickness burn and treated with 10% rhubarb decoction 12 ml/kg, once every 6 hrs. The animals were killed 24 hrs after burn, the intestinal juice was collected after intestinal lavage, and centrifuged for determination of IgA, total protein, C3 and high density lipoprotein. RESULTS: IgA content in intestinal juice of Group A, B, C and D were 93.5 +/- 13.7 mg/L, 150.8 +/- 44.6 mg/L, 43.4 +/- 21.5 mg/L and 59.8 +/- 19.3 mg/L respectively, suggesting that it was significantly reduced by burn (P < 0.05), while rhubarb decoction could increase it in either healthy or burn mice (P < 0.05). The other immune associated substance, such as total protein, C3 and high density lipoprotein in intestinal juice were also increased significantly after rhubarb decoction treatment. CONCLUSION: Rhubarb could increase the reduced intestinal juice IgA content in mice caused by burn, it may be an important mechanism of rhubarb in protecting muco-membranous barrier.

[Studies on metabolites of baicalin in human urine].

OBJECTIVE: We are interested in the metabolism of baicalin in human body to offer more information to clinic. METHOD: Urine of ten persons was collected during 2-18 h after taking 1.0 g of baicalin orally, and the urine was concentrated to 0.5 L, then filtered, the solution being loaded on an iron exchange column eluted with 1.0 L of H2O and 1.0 L of 50% EtOH. The EtOH fraction was concentrated and loaded on a Sephadex LH 20 column eluted with 0.5 L of H2O to obtain 15 fractions, which were detected by silica gel TLC (CHCl3-MeOH-H2O, 1:1:0.1). The fractions 8-11 showed flavonoid characteristics by spraying FeCl3 reagent, and the four fractions were combined to obtain the sample containing metabolites of baicalin for HPLC analysis and LCMS measurement. RESULTS AND CONCLUSIONS: M1, M2 and M3 was identified as baicalein-6-O-beta-glucopyranuronoside; 6-O-methyl-baicalin-7-O-beta-glucopyranuronoside and baicalin.

Effects of muscle relaxants on EEG, ABR and EMG in rabbits.

The effects of baclofen and pancuronium bromide on evoked electromyogram (EMG), cortical electroencephalogram (EEG) and auditory brainstem responses (ABR) were studied in pentobarbital anesthetized normal rabbits. Evoked EMG was measured in the gastrocnemius muscle by electrical stimulation of the sciatic nerve. Intravenous injection of baclofen decreased EEG and arterial blood pressure and light reflex, however, it had no significant influence on EMG or ABR at doses of 10 and 20 mg/kg/h. Pancuronium bromide immediately inhibited respiration, decreased EEG and EMG, however, it had no significant influence on arterial blood pressure, ABR, or light reflex, at doses of 0.4 and 1.0 mg/kg/h in anesthetized rabbits. ABR waves were observed until just before cardiac arrest with both of the muscle relaxants. It is suggested that ABR are not influenced by central or peripheral muscle relaxants, or by pentobarbital.

Cyclopeptides from Stellaria yunnanensis.

In a previous paper, we reported the structural elucidation of stellarin A, a new cyclic heptapeptide, from the fresh roots of Stellarina yunnanensis Franch. Further chemical study on this plant led to the isolation of another two new cyclopeptides named stellarin B and C. Their structures were established to be cyclo(Gly-Ser-HOIle-Phe-Phe-Ala) and cyclo(Gly-Ser-HOIle-Phe-Phe-Ser), respectively, by spectral methods.

Enhancement of natural killer cell activity in human immunodeficiency virus-infected subjects by in vitro treatment with biologic response modifier OK-432.

A decrease in natural killer (NK) cell function has been related to the progression of human immunodeficiency virus (HIV) infection. In the present study, we assessed the ability of a streptococcus-derived biologic response modifier, OK-432, to augment NK lysis of uninfected K562 and U937 cells and HIV-infected U937 cells by peripheral blood mononuclear cells (PBMC) from HIV-seropositive homosexual men. Optimal two- to fourfold increases in lysis of the three targets were observed after pretreatment of PBMC from HIV-negative subjects for 4 h with 2 micrograms of OK-432 per ml. This effect was related primarily to gamma interferon (IFN-gamma) production induced by OK-432 and was not linked to production of tumor necrosis factors alpha and beta or to monocytes in the cultures. The enhancing effect of OK-432 on NK cell function was diminished but still evident in PBMC from subjects with relatively early-phase (< 3-year) HIV infection and high CD4+ cell counts and was lower in subjects with longer-term HIV infection (> 3 years), in association with reduced production of IFN-gamma. Augmentation of NK cell activity in HIV-infected men by OK-432 was comparable to that induced by treatment of cells with 1,000 U of IFN-alpha or interleukin 2 per ml. The data suggest that the NK cell-enhancing effects of OK-432 are at least in part mediated by IFN-gamma and that OK-432 may be effective in treatment of patients with early-phase HIV infection.

[Changes in adhesion features of leukocytes in rats with severe burns].

Twenty-four rats were divided into 3 groups: normal controls (8), simple burned (8), burned plus Polygonum cuspidatum (p.c.)-treated (8). Three hours after burn, 3ml of whole blood was drawn to isolate polymorphonuclear neutrophilic leucocytes (PMNs). Confluent monolayer of rat microvessel endothelial cells was grown in a parallel plate of flow chamber. PMNs were driven through the flow chamber under different wall shear stresses. Leukocyte-endothelia adhesion was observed with an Olympus inverted microscope and recorded with a colour microscopic TV set. The adhesive characteristic curve of leukocyte was significantly different between the normal and burned groups. The number of PMNs adhesive to endothelium increased obviously 3 hours after burn. Under 0.196 Pa of wall shear stress, the adhesion rate of PMNs was 81.2 + 4.1% in the burned group and 58.8 + 4.6% in the normal group. The PNMs adhesion rate was decreased to near normal in p.c.-treated group. The average survival time in the burned group was 4.1 + 1.2 hours, much lower than that in the p.c. treated group (33.2 + 15.8 hours). However, in the simple burned group, one rat survived for 24 hours and its PMNs adhesive curve was similar to that of the normal or p.c.-treated groups. The findings indicate that PMNs adhesion features increase apparently after severe burn, which is closely related to the survival rate.

[Studies on water soluble polysaccharides isolated from Tribulus terrestris L--purification and preliminary structural determination of heteropolysaccharide H].

Crude polysaccharides extracted from the stem and leaf of Tribulus terristris L after the removal of crude saponins are a mixture of heteropolysaccharides composed of Ara, Rha, Xyl, GalA, Gal, Glc and Man in molar ratios of 6.0:2.1:1:3.6:3.4:7.7:2.9. A homogeneous polysaccharide H obtained by gradation and purification contains Ara, Rha, Xyl, GalA. Gal and Glc. in molar ratios of 1.6:2.4:0.1:3.5:1.3:1. Its molecular weight was found to be 1 x 10(5). By means of pectinase and beta-D-galactosidase enzymolysis, periodate oxidation, Smith degradation, partial hydrolysis with acid, methylation, GC and GC-MS, the H contains alpha-D-GalA (1-4) and L-Rha (1-2) probably linked alternately as main chain with some L-Rha (1-2) side chains.

[A new diterpenoid quinone sapriparaquinone].

A new diterpenoid quinone, named sapriparaquinone (1) was isolated from the root of Salvia prionitis Hance. 1 was identified as 3-hydroxy-6-methyl-2-(1-methyl)-5-(4-methylpenten-3-yl)-1, 4-naphthoquinone by spectral analysis and chemical transformation of saprorthquinone (2). 1 and 2 are 4,5-seco-5, 10-frideoabietane derivatives and must be biogenetically derived from an abietane structure. Like 2, 1 also showed cytotoxicity against P388 leukemia cells.

Chronotropic Effect of the Methanolic Extracts of the Plants of the Paris Species and Steroidal Glycosides Isolated from P. vietnamensis on Spontaneous Beating of Myocardial Cells1.

Rhizomes of five identified plants of the PARIS species, Liliaceae, and Rhizoma Paridis which are sold as a crude drug named "Zao Xiu", "Qiyeyizhihua" or other names in nine different markets in China were tested for their effects on cultured cardiomyocytes. In the standard medium, eight methanol extracts out of sixteen at a concentration of 0.2 mg/ml stopped the spontaneous beating of myocardial cell sheets, but these extracts significantly increased the beating rate when the concentration was reduced to one half. In the culture medium with a low calcium concentration, 0.5 mM, the beating rate of the cells decreased to about 60% of that of the control in the standard medium. The addition of five of the extracts to the low calcium medium at a concentration of 0.1 mg/ml caused a stop of cell beating, but the other extracts increased beating rate at least by 10%. Three steroidal glycosides isolated from the rhizomes of P. VIETNAMENSIS (Takht.) H. Li also stimulated cell beating. Among them, diosgenin-3- O-alpha- L-rhamnopyranosyl-(1-->2) - (alpha- L-arabinofuranosyl-(1-->4))- D-glucopranoside (compound 1) was the most effective stimulant for cell beating as well as calcium uptake by the myocardial cells.

Diterpenoids from Salvia prionitis.

Seven diterpenoids, 3-ketosapriparaquinone ( 1), salvilenone, 3-hydroxysalvilenone ( 2), arucadiol, royleanone, sugiol, and ferruginol were isolated from the root of SALVIA PRIONITIS Hance (Labiatae); Compounds 1 and 2 are new natural products. The structure of 1 was established through the interpretation of spectral data.