[Maternal periconceptional folic acid supplementation and its effects on the prevalence of fetal neural tube defects].

OBJECTIVE: To find out the status of folic acid supplementation among women, to evaluate the prevention effects on neural tube defects (NTDs), and to explore the factors impact on folic acid supplementation compliance. METHODS: Based on the routine data of 92 121 women in prenatal health care and birth defect surveillance system in Tongzhou District of Beijing from 2013 to 2018, we described the prevalence of periconceptional folic acid supplementation, pre-pregnancy folic acid supplementation and regularly folic acid supplementation (compliance supplementation). Trend χ2 tests were used to evaluate the change of folic acid supplementation prevalence. The prevalence difference among the women with folic acid supplementation and without supplementation were tested with Fisher's exact test. Factors asso-ciated with folic acid supplementation compliance rate were analyzed with univariate and multivariate Logistic regression model. RESULTS: The prevalence of periconceptional folic acid supplementation during the six years was 90.08% and it was increased from 2013 to 2018, but the rate of pre-pregnancy and regular supplementation was only 41.5% and declined from 2013 to 2018, especially 2013 to 2015. The prevalence of NTDs among the fetuses whose mothers took folic acid during periconceptional period was 5.5/10 000, while the prevalence for the fetuses whose mothers did not take folic acid was 19.7/10 000 (P < 0.001), the rates ratio was 27.9% (χ2=23.74, P < 0.001). The difference between the prevalence of NTDs among the fetuses whose mothers took folic acid only and multiple micronutrients was not statistically significant. After controlling the confounding factors, it was found that the compliant folic acid supplementation rates in women, whose household registrations were outside Beijing and whose education levels were junior high school or below, and who were younger than 25 years old, and who were multiparas and who were pre-pregnancy underweight or obese, were lower than those of the corresponding control groups (P < 0.05). CONCLUSION: The rate of folic acid supplementation among women in Tongzhou District of Beijing was relatively high, but their compliance was poor. Women who did not take folic acid during periconception seriously affected the prevention effect of NTDs. We should focus on women who are younger than 25 years old, lower educated, pre-pregnancy underweight or obese, multiparas and nonlocal household registers, in order to improve the periconceptional folic acid supplementation compliance and improve the effects of NTDs prevention.

Patient-derived organoids of non-small cells lung cancer and their application for drug screening.

Patient-derived organoids (PDOs) are emerging as preclinical models with promising values in personalized cancer therapy. The purpose of this study was to establish a living biobank of PDOs from patients with non-small cell lung cancer (NSCLC) and to study the responses of PDOs to drugs. PDOs derived from NSCLC were cultured in vitro, and then treated with natural compounds including chelerythrine chloride, cantharidin, harmine, berberine and betaine with series of concentrations (0.5-30 µM) for drug screening. Phenotypic features and treatment responses of established PDOs were reported. Cell lines (H1299, H460 and H1650) were used for drug screening. We successfully established a living NSCLC organoids biobank of 10 patients, which showed similar pathological features with primary tumors. Nine of the 10 patients showed mutations in EGFR. Natural compounds chelerythrine chloride, cantharidin and harmine showed anticancer activity on PDOs and cell lines. There was no significant difference in the 95% confidence interval (CI) for the IC50 value of chelerythrine chloride between PDOs (1.56-2.88 µM) and cell lines (1.45-3.73 µM, p>0.05). PDOs were sensitive to berberine (95% CI, 0.092-1.55 µM), whereas cell lines showed a resistance (95% CI, 46.57-2275 µM, p<0.0001). PDOs had a higher IC50 value of cantharidin, and a lower IC50 value of harmine than cell lines (p<0.05, 7.50-10.45 µM and 4.27-6.50 µM in PDOs, 3.07-4.44 µM and 4.69-544.99 µM in cell lines, respectively). Both PDOs and cell lines were resistant to betaine. Chelerythrine chloride showed the highest inhibitory effect in both models. Our study established a living biobank of PDOs from NSCLC patients, which might be used for high-throughput drug screening and for promising personalized therapy design.

[Recurrent syncope related to catecholaminergic polymorphic ventricular tachycardia due to de novo RyR2-R2401H mutation].

Objective: To explore the clinical and molecular genetic features of a Chinese patient with catecholaminergic polymorphic ventricular tachycardia (CPVT). Methods: Clinical data including resting electrocardiography, echocardiography and treadmill exercise testing of a patient with CPVT admitted to our department in March 2013 were analyzed, and the peripheral venous blood samples of the patient and his family members and 400 ethnicity-matched healthy controls were obtained. All exons and exon-intron boundaries of the six CPVT-related genes including RYR2, CASQ2, TRDN, CALM1, KCNJ2 and ANKB were sequenced to detect the variants related to CPVT. The relationship between the genotypes and phenotypes was analyzed to direct the target therapy. Results: Recurrent syncope induced either by exercise or extreme frightened fear was observed in this patient. There was no positive family history of syncope or sudden death. The resting electrocardiography and echocardiography of the patient were normal, while the exercise testing revealed bidirectional and polymorphic ventricular tachycardia. A cardiac ryanodine receptor gene mutation (R2401H) was identified in this patient, while this mutation was absent in his parents and sister and 400 controls. No variant was detected in the remaining five candidate genes. Treatment with high dose of metoprolol succinate (118.75 mg/d) was effective and patient was free of syncopal attack during the 2 years follow-up. Conclusion: This is the first report on RyR2-R2401H mutation in Chinese patient with CPVT, and high dose of metoptolol is the effective therapy option for CPVT related to RyR2 mutation.

Effects of tryptophan supplementation on growth performance, antioxidative activity, and meat quality of ducks under high stocking density.

High stocking density (STD) could affect duck welfare and production. The objective of our study was to investigate whether dietary tryptophan (TRP) supplementation could alleviate the detrimental effects of high STD on ducks. White Pekin ducks at 4 to 6 wk of age were raised at 11 birds/m(2) and fed diets containing 0.18, 0.48, 0.78, or 1.08% TRP for 21 d. Growth performance, concentrations of TRP and metabolites in the blood and hypothalamus, antioxidative activities in serum and tissue, meat quality, serum uric acid, and urea nitrogen were measured. Weight gain and feed efficiency were significantly improved by TRP supplementation at ≥ 0.48 and ≥ 0.78% (P < 0.05 and P < 0.001, respectively). Serum TRP, hypothalamic TRP, 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacitic acid (5-HIAA), and 5-HIAA/5-HT were also increased significantly (P < 0.01). These increases plateaued at 0.48% TRP, and no further improvement was obtained by adding more TRP to the diet. Dietary TRP supplementation significantly increased levels of total antioxidant capacity, glutathione peroxidase (GSH-Px), and catalase (CAT) in serum; GSH-Px in liver; and GSH-Px and CAT in breast muscle (P < 0.05). Malondialdehyde levels in breast muscle decreased (P < 0.001). Drip loss of breast muscle and pH decline at 45 min postmortem were reduced by TRP supplementation (P < 0.01 and P < 0.05, respectively). Meat color was similar among different treatments (P > 0.05). Breast muscle shear force was increased significantly when dietary TRP level increased to 1.08% (P < 0.01). For ducks raised at 11 birds/m², dietary TRP supplementation could alleviate stress and improve growth performance, antioxidative activity, and meat quality.

Early third trimester maternal response to glucose challenge and pregnancy outcome in Chinese women-relationship between upper distribution level and recommended diagnostic criteria.

BACKGROUND/OBJECTIVES: The objective of this study was to examine the relationship between upper distribution levels of glucose values in the 75-g oral glucose tolerance test (OGTT) and recommended diagnostic criteria for gestational diabetes mellitus (GDM) and adverse pregnancy outcomes. SUBJECTS/METHODS: The distribution of the OGTT 2-h values of 13,501 pregnant women, which were below the World Health Organization (WHO) threshold for overt diabetes mellitus (DM), and managed in one teaching hospital in China, was reviewed and related to maternal characteristics and pregnancy outcomes. RESULTS: For the entire group, the 90th and 95th percentile values of the OGTT 2-h glucose level, respectively, were close to the diagnostic cutoff values of the WHO and International Association of Diabetes and Pregnancy Study Groups (IADPSG) criteria. For adverse maternal outcomes, glucose level above the 90th percentile value was associated with increased hypertensive disorders, whereas no difference was seen with cutoff using the 95th percentile value. For perinatal outcomes, the 90th percentile was associated with increased neonatal intensive care unit admission and hypoglycemia, whereas the 95th percentile showed in addition association with phototherapy for jaundice and 5th-minute Apgar score <7. Although no differences in the incidence of adverse pregnancy outcomes were found using the different cutoffs, the >95th percentile cutoff value would have missed out 33.3-56.7% of the cases of adverse outcomes that would otherwise have been attributed to GDM. CONCLUSIONS: Further studies are warranted to clarify which diagnostic criterion is most appropriate universally to identify adverse pregnancy outcomes attributed to GDM, and which could be mitigated with treatment specific for GDM.

Induction of striatal neurogenesis enhances functional recovery in an adult animal model of neonatal hypoxic-ischemic brain injury.

While intraventricular administration of epidermal growth factor (EGF) expands the proliferation of neural stem/progenitor cells in the subventricular zone (SVZ), overexpression of brain-derived neurotrophic factor (BDNF) is particularly effective in enhancing striatal neurogenesis. We assessed the induction of striatal neurogenesis and consequent functional recovery after chronic infusion of BDNF and EGF in an adult animal model of neonatal hypoxic-ischemic (HI) brain injury. Permanent brain damage was induced in CD-1 (ICR) mice (P7) by applying the ligation of unilateral carotid artery and hypoxic condition. At 6 weeks of age, the mice were randomly assigned to groups receiving a continuous 2-week infusion of one of the following treatments into the ventricle: BDNF, EGF, BDNF/EGF, or phosphate buffered saline (PBS). Two weeks after treatment, immunohistochemical analysis revealed an increase in the number of BrdU(+) cells in the SVZ and striata of BDNF/EGF-treated mice. The number of new neurons co-stained with BrdU and betaIII-tubulin was also significantly increased in the neostriata of BDNF/EGF-treated mice, compared with PBS group. In addition, the newly generated cells were expressed as migrating neuroblasts labeled with PSA-NCAM or doublecortin in the SVZ and the ventricular side of neostriata. The new striatal neurons were also differentiated as mature neurons co-labeled with BrdU(+)/NeuN(+). When evaluated post-surgical 8 weeks, BDNF/EGF-treated mice exhibited significantly longer rotarod latencies at constant speed (48 rpm) and under accelerating condition (4-80 rpm), relative to PBS and untreated controls. In the forelimb-use asymmetry test, BDNF/EGF-treated mice showed significant improvement in the use of the contralateral forelimb. In contrast, this BDNF/EGF-associated functional recovery was abolished in mice receiving a co-infusion of 2% cytosine-b-d-arabinofuranoside (Ara-C), a mitotic inhibitor. Induction of striatal neurogenesis by the intraventricular administration of BDNF and EGF promoted functional recovery in an adult animal model of neonatal HI brain injury. The effect of Ara-C to completely block functional recovery indicates that the effect may be the result of newly generated neurons. Therefore, this treatment may offer a promising strategy for the restoration of motor function for adults with cerebral palsy (CP).

The alkaloid Berberine inhibits the growth of Anoikis-resistant MCF-7 and MDA-MB-231 breast cancer cell lines by inducing cell cycle arrest.

Berberine is a pure phenanthren alkaloid isolated from the roots and bark of herbal plants such as Berberis, Hydrastis canadensis and Coptis chinensis. Berberine has been established to inhibit the growth of breast cancer cells, but its effects on the drug resistance and anoikis-resistance of breast cancer cells have yet to be elucidated. Anoikis, or detachment-induced apoptosis, may prevent cancer progression and metastasis by blocking signals necessary for survival of localized cancer cells. Resistance to anoikis is regarded as a prerequisite for metastasis; however, little is known about the role of berberine in anoikis-resistance. We established anoikis-resistant cells from the breast cancer cell lines MCF-7 and MDA-MB-231 by culturing them on a Poly-Hema substratum. We then investigated the effects of berberine on the growth of these cells. The anoikis-resistant cells had a reduced growth rate and were more invasive than their respective adherent cell lines. The effect of berberine on growth was compared to that of doxorubicine, which is a drug commonly used to treat breast cancer, in both the adherent and anoikis-resistant cell lines. Berberine promoted the growth inhibition of anoikis-resistant cells to a greater extent than doxorubicine treatment. Treatment with berberine-induced cell cycle arrest at G0/G1 in the anoikis-resistant MCF-7 and MDA-MB-231 cells as compared to untreated control cells. In summary, these results revealed that berberine can efficiently inhibit growth by inducing cell cycle arrest in anoikis-resistant MCF-7 and MDA-MB-231 cells. Further analysis of these phenotypes is essential for understanding the effect of berberine on anoikis-resistant breast cancer cells, which would be relevant for the therapeutic targeting of breast cancer metastasis.

Induction of quinone reductase by a methanol extract of Scutellaria baicalensis and its flavonoids in murine Hepa 1c1c7 cells.

The effect of extracts of scutellariae radix (Scutellaria baicalensis Georgi) and its flavonoids, baicalin, baicalein and wogonin, on induction of quinone reductase (QR) in the Hepa 1c1c7 murine hepatoma cell line was examined. A significant and dose-dependent induction of QR activity was observed in the methanol extract of scutellariae radix and baicalin. HPCL analysis showed that baicalin was contained as a main component in the methanol extract of scutellariae radix, indicating that baicalin may be the major active principle of QR induction mediated by scutellariae radix extract. To elucidate the mechanism of baicalin-mediated induction of QR enzyme activity, the effect on QR mRNA levels in Hepa 1c1c7 cell cultures was investigated. Using reverse transcriptase-polymerase chain reaction techniques, time- and dose-dependent induction of QR mRNA levels by baicalin were demonstrated in Hepa 1c1c7 cells. On the basis of these results, the scutellariae radix extract or baicalin can be regarded as a readily available, promising, novel cancer chemopreventive agent.

[Haemorrheological effects of different processed products of radix Aconiti Kusnezoffii on animal model with syndrome of "accumulation of cold and blood stasis"].

The effects of Radix Aconiti Kusnezoffii processed by moistening in combination with steaming method and by pharmacopoeial method as well as of the crude drug on haemorrheological nature and the plasma fibronogen content and platelet aggregation rate in the rat model with syndrome of "accumulation of cold and blood stasis" were studied, the result shows that the product processed by moistening in combination with steaming works best.

Effects of clonidine on the calcium content and morphology of rat salivary glands.

These effects were examined with and without pretreatment of animals with reserpine and the adrenergic antagonists prazosin (alpha 1), yohimbine (alpha 2) and propranolol (beta). The effects of clonidine on glandular concentrations of norepinephrine and dopamine also were examined. These effects were compared with those of xylazine, a presynaptic alpha 2-adrenergic agonist. A single, high dose of clonidine followed by an overnight fast caused marked increases in calcium content and acinar secretory granules in the submandibular and sublingual glands, similar to those caused by reserpine. However, the calcium content of the parotid gland was not altered by clonidine, although there seemed to be a modest increase in acinar secretory granules. The clonidine-induced increase in submandibular calcium content could not be attributed to any adrenergic receptor activity since it was not blocked by either alpha- or beta-adrenergic antagonists. Unlike reserpine, clonidine did not affect catecholamine concentrations in the parotid and submandibular glands. Pretreatment with reserpine did not significantly alter the clonidine-induced increase in submandibular calcium content. It is likely that the greater accumulation of acinar secretory granules is related to the increased calcium stores of the glands in clonidine- and/or reserpine-treated rats. The large differences in calcium content among the three glands might be attributable, in part, to differences in the calcium-binding capacity of their secretory granules. Possible mechanisms for the clonidine effects on salivary-gland calcium include disturbances in membrane-associated pools or gating mechanisms for calcium, which need further study.

Secretory responses to autonomic stimulation of rat salivary glands following reserpine treatment.

The response to stimulation of the parasympathetic innervation to parotid or submandibular glands of reserpinized rats was altered from that of untreated rats. Thus, acute reserpinization, like other types of sympathectomy, resulted in increase in volume of parasympathetically-evoked parotid or submandibular saliva when comparison was made with evoked saliva from untreated glands. As norepinephrine is depleted by reserpine, there was no response to stimulation of sympathetic nerves to these reserpinized glands. Adrenergic receptors were normally activated by administration of autonomic agonists. Thus a single high dose of reserpine can cause the same effects as those induced by chronic administration of low doses of reserpine, i.e. a 3-fold increase in calcium (Ca) concentration of submandibular gland but no change in Ca concentration of parotid gland. Although sympathetic stimulation caused no change in Ca concentration of submandibular or parotid glands of reserpine (acute)-treated rats, stimulation with isoproterenol (25 mg/kg, i.p., 60 min) produced a 32-35 per cent decrease in glandular Ca concentration from that of unstimulated reserpinized glands. Glands of untreated rats showed a 52 per cent depletion after 60 min of isoproterenol stimulation; however, Ca output in parotid saliva from reserpinized rat for 60 min of stimulation was not changed from that of untreated rats, but that of submandibular saliva was two times greater. Ca concentration of submandibular saliva was unchanged during 60 min-stimulation of reserpine-treated rats, but that of untreated rats decreased.

Calcium concentration of salivary glands and saliva after chemical or surgical sympathectomy.

Calcium concentration of the submaxillary (SM) gland of adult rat was increased to 2-3 times control levels 24 h after a single high dose of reserpine (RES) (5 mg/kg body weight), 6-hydroxydopamine (6-OHDA) (20 mg/kg body weight), or surgical removal of a superior cervical ganglion (Sx). The increase could also be induced within 24 h after a single injection of lower doses of RES (0.05, 0.5 mg/kg body weight). Increase in calcium concentration in the parotid (PA) was not found under any of the above conditions, but did occur transiently at times within 24 h. The increase in glandular calcium concentration was found to be temporally related to depletion of norepinephrine (NE). The time frame for initiation of calcium changes was not the same for chemical and surgical sympathectomy (Sx). Increase in calcium concentration occurred after onset of sympathectomy-induced degeneration secretion; the onset varied with the kind of sympathectomy, and was seen 1 h after drug administration with either RES or 6-OHDA, but not until about 13 h after surgical Sx. Similarly, NE was markedly depleted, but later with surgical than with chemical Sx. Furthermore, with both kinds of sympathectomy, depletion of NE occurred earlier with PA than with SM, since normal basal levels of NE of SM are 3 times as great (3400 ng/g wet weight), compared with PA levels (1300 ng/g wet weight). The fact that NED was the neurotransmitter eliciting the degeneration secretion was confirmed physiologically by examination of the composition of this secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of cocaine and desmethylimipramine on the uptake, retention and metabolism of 3H-5-hydroxytryptamine in rat brain slices.

Uptake and metabolism of 3H-5-dydroxytryptamine (3H-5-HT) in slices of hypothalamus, brain stem caudate nucleus and parietal cortex were examined in vitro. The results indicate that 3H-5-HT taken up by neurons is firmly retained and that there are regional differences in the uptake and retention of 3H-5-HT. For example, the caudate had a significantly greater maximum rate of uptake and binding (Vmax) than the other brain areas tested, particularly the cortex. In contrast the caudate appeared to have the lowest apparent for 3H-5-HT uptake (Km). Uptake and retention of 3H-5-HT by four brain areas were markedly inhibited by cocaine and desmethylimipramine (DMI) in vitro. The inhibitory effect of both drugs was noncompetitive in nature, with exception of thct that cocaine and DMI failed to reduce the formation of 3H-5-hydroxy-indoleacetic acid despite ther potent blocking action of 3H-5-HT uptake suggests that a significant amount of 3H-5-HT is oxidatively deaminated at extraneouronal sites. This extraneuronal metabolism of 5-hydroxytryptamine may play an play an important role in regulating the action of this neurotransmitter.