A prospective multicentre controlled study of Gaoweikang (Chinese multiherb extract-based tincture) used in high-risk HPV infections.

OBJECTIVE: The aim of the study was to investigate the safety and antiviral efficacy of a Chinese multiherb extract-based tincture (GWK) on a population of patients with high-risk human papilloma (hrHPV) infections and hrHPV-caused cervical low-grade squamous intraepithelial lesions (LSILs). PATIENTS AND METHODS: Patients with persistent hrHPV infection were enrolled in Group A, including A1 subjects, who received the intervention, and A2 subjects, who received the control. Patients with hrHPV infection causing cervical LSIL were enrolled in Group B, which included B1 subjects, who received the intervention, and B2 subjects, who served as the control. For Groups A1 and B1, hrHPV was tested at 3 months (M3) and 6 months (M6) after the intervention. The side effects were also analyzed. RESULTS: At baseline (D0), a total of 99 patients were enrolled in Group A, with 50 subjects in Group A1 and 49 subjects in Group A2. A total of 91 patients were enrolled in Group B, with 45 subjects in Group B1 and 46 subjects in Group B2. There was no significant difference in the characteristics, including average age, age stratification, and HPV genotype. At M6, both Group A1 and Group B1 had a higher hrHPV clearance rate than the control group (A1/A2: 80.0% vs. 20.4%; B1/B2: 64.4% vs. 15.2%, p<0.001). At M6, the effective rates of Group A1 and Group B1 were 84% (42/50) and 68.9% (31/45), respectively. The side effect rates of Groups A1 and B1 were 11.5% (6/52) and 11.1% (5/45), respectively. Most adverse reactions involved local discomfort, including vulvar erythema, vulvar itch, increased vaginal discharge, cervical bleeding, and mild pain in the lower abdomen. Univariate logistic regression analysis showed that the intervention had an OR of 12 (95% CI 4.431-32.50) for clearing persistent HPV infection (p<0.001). For cervical LSIL, the intervention had an OR of 10.1 for clearing persistent HPV infection (95% CI 3.68-27.7) (p<0.001). CONCLUSIONS: The results of this study suggest that the Chinese multiherb extract-based tincture GWK is safe and well tolerated. Furthermore, this preliminary study showed that this Chinese multiherb extract-based tincture is helpful for promoting HPV clearance in cases of persistent HPV and HPV-induced LSIL.

[Neuroprotective effect of ginsenoside Rb-1 on a rat model of Alzheimer's disease].

Objective: To investigate the protective mechanism of ginsenoside Rb-1 on the brain in a rat model of Alzheimer's disease. Methods: Fifty-six male Sprague-Dawley rats were randomly divided into control group, model group, low-dose Rb-1 group (Rb-1: 25 mg•kg(-1)•d(-1)) and high-dose Rb-1 group (Rb-1:50 mg•kg(-1)•d(-1)). Morris water maze was designed to observe the changes of learning and memory ability in rats. Flow cytometry was used to detect the apoptosis of hippocampal neurons. Immunohistochemistry and Western blot were employed to detect the expression levels of apoptosis-related genes (p53, Bax, cytochrome C (Cyto C), Caspase-3 and caspase-9) and anti-oxidative stress-associated genes (nuclear Factor-E2-related factor 2 (Nrf2), kelch-like ECH-associated protein 1 (keap-1), heme oxygenase 1(HO-1) and NADPH quinone dehydrogenase 1 (NQO1)).The activities of catalase (CAT), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were detected by relevant kits. ANOVA and Tukey-Kramer test were used for statistical analysis. Results: The learning and memory ability of rats in the model group was lower than that of the control group (P<0.01).The learning and memory ability of rats in the high-dose Rb-1 treatment group was significantly higher than that of the model group [(80±8) s vs (100±11) s, t=5.390, P<0.01]. The expression levels of apoptosis-related genes (p53, Bax, Cyto C, caspase-3 and caspase-9) in the model group were significantly higher than those in the control group (P<0.01), while the expression levels of these genes in low-dose and high-dose Rb-1 groups were significantly lower than those of the model group (P<0.01). The expression levels of Nrf2, HO-1 and NQO1 genes in the model group were significantly lower than those in the control group (P<0.05), while the expression of these genes in low-dose and high-dose Rb-1 groupswere significantly higher than those of the model group (P<0.01). The activities of CAT, GSH-Px and SOD in the model group were lower than those in the control group (P<0.01), however the activities of CAT, GSH-Px and SOD in low-dose and high-dose Rb-1 groups were higher than those of model group (P<0.05). Conclusions: Both low-dose and high-dose Rb-1 have protective effect on memory and cognitive function of Alzheimer's disease rats by reducing the damage and apoptosis of hippocampal neurons, down-regulating the expression levels of p53, Bax, Cyto C, caspase-3 and caspase-9, up-regulating the expression of Nrf2, HO-1 and NQO1 genes, and increasing the activities of CAT, GSH-Px and SOD. Moreover, the protective effect of Rb-1 on rat brain may be dose-dependent.

[Study on literatures of health preservation and practitioners in Taiping Yulan: Fangshu].

Taiping Yulan(, Imperial overview from the Taiping reign or Readings of the Taiping Era) was a large encyclopedia which compiled by Li Fang in the early Northern Song Dynasty. Its content was broad and the resources of quotation were varied. It contained a large number of medical knowledges. Among this book, the "Fangshu" volume consisted of 18 parts, the first five volumes of health and medical recorded 4 methods of health preservation (nurturing vitality), 88 practitioners, cited 59 kinds of literatures and 160 historical materials. It has important academic value to the study of ancient health preservation and inheritance development of medical school.

[Effects of combined application of culture supernatant of human umbilical cord mesenchymal stem cells and ciprofloxacin on Staphylococcus aureus in vitro].

Objective: To explore the effects of combined application of culture supernatant of human umbilical cord mesenchymal stem cells (hUCMSCs) and ciprofloxacin on Staphylococcus aureus (SA) in vitro. Methods: hUCMSCs were isolated from umbilical cord tissue of full-term healthy fetus after cesarean section and cultured. Cells in the third passage were used in the experiments after identification. SA strains isolated from wounds of burn patients in our burn wards were used in the experiments. Cells were divided into 0, 10, 100, and 1 000 ng/mL lipopolysaccharide (LPS) groups according to the random number table (the same dividing method below). Cells were cultured with culture medium of mesenchymal stem cells (MSCs) after being treated with medium containing the corresponding mass concentrations of LPS for 12 h. At post culture hour (PCH) 6, 12, and 24, 6 wells of culture supernatant of cells in each group were obtained to measure the content of LL-37 with enzyme-linked immunosorbent assay. Ninety blood agar plates were divided into ciprofloxacin control group (CC), ciprofloxacin+ supernatant group (CS), and ciprofloxacin+ supernatant+ LL-37 antibody group (CSL), with 30 blood agar plates in each group. Blood agar plates in group CC were coated with 1.5×10(8) colony forming unit (CFU)/mL bacteria solution prepared with normal saline. Blood agar plates in group CS were coated with 1.5×10(8) CFU/mL bacteria solution prepared with normal saline and culture supernatant of hUCMSCs (cultured by culture medium of MSCs, the same below) in double volume of normal saline. Blood agar plates in group CSL were coated with 1.5×10(8) CFU/mL bacteria solution prepared with normal saline, culture supernatant of hUCMSCs in double volume of normal saline, and 2.6 µL LL-37 antibody in the concentration of 2 µg/mL. At PCH 12, 24, and 48, 10 blood agar plates of each group were harvested to observe the distribution of SA colony on blood agar plate and to measure the diameter of bacterial inhibition ring of ciprofloxacin. The minimum inhibitory concentration (MIC) of ciprofloxacin against SA of each group was recorded. Fractional inhibitory concentration (FIC) indexes of ciprofloxacin in groups CS and CSL at PCH 12, 24, and 48 were calculated, and the effect of synergy was evaluated. Data were processed with analysis of variance of factorial design, one-way analysis of variance, LSD-t test, Kruskal-Wallis test, and Mann-Whitney U test. Results: (1) At each PCH, the content of LL-37 in culture supernatant of cells in 10, 100, and 1 000 ng/mL LPS groups was higher than that in 0 ng/mL LPS group (with t values from 11.22 to 33.36, P values below 0.01); the content of LL-37 in culture supernatant of cells in 100 and 1 000 ng/mL LPS groups was higher than that in 10 ng/mL LPS group (with t values from 2.24 to 18.73, P<0.05 or P<0.01); the content of LL-37 in culture supernatant of cells in 1 000 ng/mL LPS group was higher than that in 100 ng/mL LPS group (with t values from 12.46 to 14.70, P values below 0.01). (2) At PCH 12, 24, and 48, the bacterial colonies in groups CC, CS, and CSL began to integrate over time. At PCH 12, 24, and 48, the diameters of bacterial inhibition ring of ciprofloxacin in group CC were 26, 24, and 23 mm, respectively, with no obvious change. At PCH 12, 24, and 48, the diameters of bacterial inhibition ring of ciprofloxacin in groups CS and CSL were 82, 71, 68 mm, and 74, 59, 56 mm, respectively, significantly longer than those of group CC. (3) At each PCH, the MIC of ciprofloxacin against SA was significantly higher in group CC than in groups CS and CSL (with Z values from 6.22 to 6.71, P values below 0.01); the MIC of ciprofloxacin against SA was significantly higher in group CSL than in group CS (with Z values all equal to 6.72, P values below 0.01). (4) FIC indexes of ciprofloxacin in groups CS and CSL at PCH 12, 24, and 48 were 0.011, 0.032, 0.032, and 0.122, 0.350, 0.350, respectively. The results indicated that culture supernatant of hUCMSCs had synergistically antibacterial effect on ciprofloxacin. Conclusions: hUCMSCs can secrete LL-37, and the secretion level is increased with increase of LPS concentration. Combination of culture supernatant of hUCMSCs and ciprofloxacin can decrease the dosage of ciprofloxacin in resisting SA. Once LL-37 is neutralized, the synergistically antibacterial effect of culture supernatant of hUCMSCs is decreased.

A noncalibration spectroscopic method to estimate ether extract and fatty acid digestibility of feed and its validation with flaxseed and field pea in pigs.

Digestibility of ether extract (EE) or fatty acids (FA) is traditionally measured by chemical analyses for EE or GLC methods for FA combined with marker concentration in diet and digesta or feces. Digestibility of EE or FA may be predicted by marker concentrations and spectral analyses of diet and digesta or feces. On the basis of Beer's law, a noncalibration spectroscopic method, which used functional group digestibility (FGD) determined with marker concentration and peak intensity of spectra of diets and undigested residues (digesta or feces), was developed to predict the apparent ileal digestibility (AID) of total FA and apparent total tract digestibility (ATTD) of EE. To validate, 4 diets containing 30% flaxseed and field pea coextruded with 4 extruder treatments and a wheat and soybean basal diet with predetermined AID of total FA and ATTD of EE were used. Samples of ingredients, diets, and freeze-dried digesta and feces were scanned on a Fourier transform infrared (FT-IR) instrument with a single-reflection attenuated total reflection (ATR) accessory. The intensity of either the methylene (CH2) antisymmetric stretching peak at 2,923 cm(-1) (R(2) = 0.90, P < 0.01) or the symmetric stretching peak at 2,852 cm(-1) (R(2) = 0.86, P < 0.01) of ingredients, diet, and digesta spectra was related strongly to the concentration of total FA. The AID of total FA of diets measured using GLC was predicted by the spectroscopic method using FGD at 2,923 and 2,852 cm(-1) (R(2) = 0.75, P < 0.01) with a bias of 0.54 (SD = 3.78%) and -1.35 (SD = 3.74%), respectively. The accumulated peak intensity in the region between 1,766 and 1,695 cm(-1) of spectra was related to EE concentration in ingredients and diets (R(2) = 0.61, P = 0.01) and feces (R(2) = 0.88, P < 0.01). The relation was improved by using second-derivative spectra of the sum of peak intensities at 1,743 and 1,710 cm(-1) for ingredients and diets (R(2) = 0.90, P = 0.01) and at 1,735 and 1,710 cm(-1) for feces (R(2) = 0.92, P < 0.01). The ATTD of EE of test diets determined with proximate analysis was estimated by the FGD of nonderivative spectra with or without baseline (R(2) = 0.90, P < 0.01) with a bias of 3.15 (SD = 3.14%) and 3.50 (SD = 3.24%), respectively. In conclusion, instead of using GLC methods or predictions based on calibrations, the AID of total FA and ATTD of EE can also be estimated directly from ATR FT-IR spectra, provided the ratio of marker in the diet and undigested residue is known.

Conjugated linoleic acid-induced milk fat reduction associated with depressed expression of lipogenic genes in lactating Holstein mammary glands.

The efficacy of conjugated linoleic acid (CLA) in diet supplements for milk fat reduction is well documented in several species. However, the mechanisms by which fatty acids regulate mammary lipogenesis remain largely unknown, especially with regard to gene expression of enzyme and regulators. In this study, 8 Holstein dairy cows in their mid-lactation period were randomly divided into 2 groups. Control cows received a Ca salt of palm oil fatty acid dietary supplement, and those in the CLA group were fed Ca salts of CLA (Ca-CLA), all in a dose of approximately 200 g∙cow(-1)∙day(-1) for 14 days. The milk yield was recorded daily, and protein, lactose, and fat in the milk were quantified every 3 days for 2 weeks. Fatty acids in the milk were analyzed with gas-liquid chromatography. Measurement of messenger RNA levels of the main lipogenic genes of lipoprotein lipase, acetyl-coenzyme A (CoA) carboxylase, fatty acid synthase, stearoyl-CoA desaturase, and transcription factors such as sterol response element binding protein 1 (SREBP1) and peroxisome proliferator-activated receptor γ was performed in biopsy samples of mammary tissue on the last day. The results indicated that dietary Ca-CLA caused a continuous reduction of milk fat (P < 0.01) with no effect on milk yield, milk protein, and lactose. The fatty acid profile in the milk from the CLA group differed from that from controls, and the yield of milk fatty acid decreased (P < 0.01) with Ca-CLA supplementation. The depressed expression of lipogenic genes (lipoprotein lipase, acetyl-CoA carboxylase, fatty acid synthase, and stearoyl-CoA desaturase) demonstrated inhibition of fatty acid de novo synthesis and uptake in the mammary gland of the CLA group. Furthermore, the gene expression of transcription factor SREBP1 was also downregulated (P < 0.01), but peroxisome proliferator-activated receptor γ was unchanged, suggesting that SREBP1 may play a key role in the regulation of lipogenic gene expression in the lactating mammary gland.

First Report of Root-Knot Nematode Meloidogyne arenaria on Atractylodis macrocephalae in China.

Atractylodis macrocephalae is an important Chinese herbal medicine plant and its rhizome is of high medicinal value. In recent years, a severe decline in yield has been observed in Bozhou City (China's largest A. macrocephalae producing area), Anhui Province, China. A survey for plant-parasitic nematodes was conducted in this area from June to September 2011. Stunted plants displayed chlorotic or necrotic lower leaves near the ground part by the growth reduction; examination of the roots of stunted plants revealed the presence of galls typical of infestation by root knot nematode. Root nodules were found on the tap and lateral roots caused the fleshy tap root deformity. The incidence of diseased plants was estimated to be 45%, and yield loss was quantified as 43.5%. Nematodes were extracted from the root samples as previously described (4) and identified by morphology, enzyme analysis, and molecular characterization. Morphology of the female perineal patterns and measurements of the second-stage juveniles (J2s) matched those of the original description of Meloidogyne arenaria. Enzyme analysis of the esterase (Est) phenotype was also typical of the AII phenotype in M. arenaria (2). DNA was extracted according to a modified protocol (1), and the rDNA-internal transcribed spacer (ITS1_5.8S_ITS2) region was amplified with universal primers V5367 (5'-TTGATTACGTCCCTGCCCTTT-3') and 26S (5'-TTTCACTCGCCGTTACTAAGG-3'). PCR yielded a fragment of 764 bp and the purified product was sequenced by Sanger's dideoxy chain termination method (ABI3730). Sequences were identical to that of M. arenaria in GenBank (Accession No. AF387092) (3). Amplification of the D2/D3 fragments of the 28S RNA with universal primers D2A (5'-ACAAGTACCGTGAGGGAAAGTTG-3') and D3B (5'-TCGGAAGGAACCAGCTACTA-3') yielded a PCR fragment of 758 bp. These sequences were also identical to that of M. arenaria in GenBank (Accession No. AF435803). For further confirmation, amplification of the IGS region with universal primers 5S (5'-TTAACTTGCCAGATCGGACG-3') and 18S (5'-TCTAATGAGGGAACCAGCTACTA-3') yielded a PCR fragment of 713 bp. These sequences were 99.64% homologous to that of M. arenaria (GenBank Accession No. MAU42342). To our knowledge, this is the first report of M. arenaria species on A. macrocephalae in China. The fleshy tap root of A. macrocephalae is the main edible part of the plant, and the damage caused by root knot nematode will potentially reduce the yield and quality of this herb. References: (1) J. L. Cenis et al. Phytopathology 83:76, 1993. (2) P. R. Esbenshade and A. C. Triantaphyllou. J. Nematol. 17:6, 1985. (3) T. C. Vrain et al. Appl Nematol. 15:563, 1992. (4) L. F. Wang et al. Forest Res. 14:484, 2001.

Carnitine and carnitine orotate affect the expression of the prolactin-releasing peptide gene.

Carnitine is involved in fatty acid metabolism in mammals and is widely used as a nutritional supplement; carnitine orotate is a more absorbable form of carnitine. We investigated the effects of carnitine and carnitine orotate on mouse prolactin-releasing peptide (PrRP) mRNA expression. Twenty-four female mice were randomly divided into four groups of six; control mice were orally drenched with physiological saline solution (250 mg/kg body weight) and treatment mice were orally drenched with carnitine (250 mg/kg) or carnitine orotate (250 or 750 mg/kg), once a day, for 20 days from parturition. The carnitine or carnitine orotate was dissolved in saline solution before administration. The hypothalamus, pituitary and ovary were sampled on day 21 after parturition, and PrRP mRNA levels in these tissues were measured by semi-quantitative PCR, with glyceraldehyde 3-phosphate dehydrogenase as a control. Expression of PrRP in mice treated with carnitine and carnitine orotate was significantly increased in the ovary and significantly reduced in the pituitary gland. Compared with the control, hypothalamus PrRP mRNA increased significantly in the carnitine and low-dose carnitine orotate groups and decreased significantly in the high-dose carnitine orotate group. We conclude that carnitine and carnitine orotate regulate expression of PrRP in the pituitary gland and ovaries.

A novel monoclonal antibody against human CD80 and its immune protection in a mouse lupus-like disease.

Blockade of the interactions between CD28/CTLA-4 and their ligands, CD80 (B7, B7.1)/CD86 (B70, B7.2), is an attractive means to induce antigen-specific peripheral tolerance in autoimmune disease and organ transplantation. In this study, we generated and characterized a monoclonal antibody (Clone 4E5) against human CD80. 4E5 could recognize both human and mouse CD80 and suppress mixed lymphocyte reaction in vitro. To investigate their potency for clinical use, we further administrated 4E5 to a mouse lupus-like disease model (C57BL/J6) induced by Pristane. 4E5 could inhibit the immune response and attenuate the severity of lupus-like disease. The data showed 4E5 function and suggested that blockade of CD80/CD28 co-stimulatory signal pathway with 4E5 is a promising strategy to decelerate the progression of lupus-like disease and other autoimmune diseases.

Effects of psoralen plus ultraviolet A irradiation on cultured epidermal cells in vitro and patients with vitiligo in vivo.

BACKGROUND: Both psoralen plus ultraviolet (UV) A (PUVA) and narrowband UVB (NB-UVB) irradiation are effective treatments for vitiligo vulgaris. However, the mechanisms of PUVA and NB-UVB in repigmentation are not thoroughly clarified. Our previous results showed that NB-UVB irradiation directly promotes melanocyte (MC) migration and stimulates MC proliferation via keratinocytes (KCs). OBJECTIVES: In the present study, we used NB-UVB as a reference for comparison to investigate the immediate effects of PUVA on MC proliferation and migration. METHODS: Cultured MCs and KCs were treated with PUVA or irradiated with NB-UVB. The direct impact of PUVA treatment on MCs was assessed in terms of its effect on MC proliferation and migration. The indirect effect of PUVA treatment and NB-UVB irradiation on MC proliferation via KCs was also investigated. The activities of matrix metalloproteinase (MMP)-2 and MMP-9, known for their influence on cell migration, were evaluated in the PUVA-treated MC and KC supernatants. The concentrations of MC mitogens/growth factors in the PUVA-treated KC supernatants were also determined. In addition, the serum levels of MC mitogens/growth factors in healthy controls, in patients with active vitiligo and in patients with repigmenting vitiligo after PUVA treatment were determined to elucidate the mechanisms of how PUVA induces vitiligo repigmentation in vivo. RESULTS: Our results demonstrated that PUVA treatment did not significantly stimulate the release of MC mitogens/growth factors from KCs. The migration of MCs was also not enhanced after PUVA treatment. The expression of MMP-2 activity in supernatants derived from PUVA-treated MCs was significantly increased as compared with the control group. However, neither MMP-2 nor MMP-9 activity in KC supernatants was stimulated by PUVA treatment. In contrast to NB-UVB, immediate effects of PUVA on MC proliferation and migration were not observed in this study. Sera from patients with repigmenting vitiligo after PUVA treatment contained higher levels of basic fibroblast growth factor, stem cell factor and hepatocyte growth factor as compared with healthy controls and patients with active vitiligo. CONCLUSIONS: Our results indicate that in addition to immune suppression, PUVA treatment creates a favourable milieu for promoting the growth of MCs in patients with vitiligo instead of directly stimulating the regrowth of MCs. Based on our results, we propose that in the active stage of vitiligo, PUVA treatment is the therapy of choice to slow down the destruction of MCs and to create a favourable environment for MCs to survive. In the stable stage of vitiligo, NB-UVB irradiation should be used to stimulate the proliferation and migration of MCs directly.

Antitumor and immunostimulating effects of Anoectochilus formosanus Hayata.

The water extract of Anoectochilus formosanus Hayata showed a potent tumor inhibitory activity in BALB/c mice after subcutaneous transplantation of CT-26 murine colon cancer cells. The tumor-inhibition ratios of mice pre-administered with A. formosanus for 2 days before tumor transplantation, and treated further for 12 consecutive days, were 55.4% and 58.9% at the oral dose of 50 and 10 mg/mouse per day, respectively. Even for the tumor-bearing mice, after oral administration of the water extract of A. formosanus for 12 consecutive days, the tumor inhibition ratios were still 23.8% and 40.5% at doses of 50 and 10 mg/mouse, respectively. Because the low-concentration water extract of A. formosanus does not show direct cytotoxicity in CT-26 tumor cells, we observed further that oral administration of the water extract of A. formosanus may activate murine immune responses, such as stimulating the proliferation of lymphoid tissues and activating the phagocytosis of peritoneal macrophages against Staphylococcus aureus. This study suggests that the antitumor activity of A. formosanus may be associated with its potent immunostimulating effect. It is worth further analyzing the immunomodulating component purified from A. formosanus, and evaluating its potential value for the treatment of human cancers.

Effects of Anoectochilus formosanus Hayata extract and glucocorticoid on lung maturation in preterm rats.

We investigated the effects of maternal administration of Anoectochilus formosanus extract and dexamethasone on lung maturation in preterm rats. A. formosanus group mothers were tube-fed A. formosanus extract (300 mg/kg body wt./day) for 7 days from days 12-18 of gestation. Dexamethasone group mothers were injected intraperitoneally with dexamethasone (0.2 mg/kg body wt.) in saline on day 18 of gestation. Control group mothers were similarly injected with saline alone. On day 19 of gestation, fetuses were delivered by cesarean section. A. formosanus treatment significantly increased the fetal lung/body weight ratio, as compared to dexamethasone treatment. Saturated phosphatidylcholine levels in fetal lung tissue and growth hormone levels in maternal serum were significantly increased in the A. formosanus- and dexamethasone-treated groups as compared to controls. The histological appearance of preterm rat lungs revealed extensive branching of intermediate airways, denser mesenchyme, and more epithelial tubules in the dexamethasone and A. formosanus groups as compared with the control group. These results suggest that antenatal A. formosanus treatment may play a role in accelerating fetal rat lung maturation.

Effect of dietary supplemental levels of vitamin A on the egg production and immune responses of heat-stressed laying hens.

Two experiments were conducted to evaluate the effect of vitamin A supplementation of a commercial layer diet on the laying performance and immune function of heat-stressed hens. In Experiment 1, two different levels of vitamin A supplementation (3,000 and 9,000 IU/kg) were used to investigate the laying performance and antibody titer against Newcastle disease virus (NDV) of heat-stressed hens. Results showed that the high level of vitamin A supplementation (9,000 IU/kg) had a beneficial effect on the feed intake and laying rate of heat-stressed hens (P < 0.05), compared with the control group (3,000 IU/kg). The antibody titers were not influenced by the level of vitamin A (P > 0.05). In Experiment 2, the effect of four levels of vitamin A (3,000, 6,000, 9,000, and 12,000 IU/kg) on the antibody titer to NDV and T lymphocyte proportion was studied. The experimental birds were exposed to a high temperature (31.5 C) 15 d after NDV vaccination (Treatment 1) or immediately (Treatment 2). The results showed that the egg weight was increased (P < 0.01) by the high levels of vitamin A supplementation (6,000 and 9,000 IU/kg), but feed intake, laying rate, and body weight loss were not (P > 0.05). In Treatment 1, vitamin A had no significant effect on antibody titers against NDV in normal or hot environments but increased (P < 0.01) the proportion of alpha-naphthyl acetate esterase (ANAE)-positive cells. Vitamin A supplementation had a significant effect on NDV antibody titer and ANAE-positive cell proportion in Treatment 2 (P < 0.01). The results of the present study suggested that vitamin A supplementation in commercial layer diets to layer chickens under heat stress was beneficial to laying performance and immune function.

[Experimental pharmacodynamic study on the anti-convulsion effect of shenpu decoction].

OBJECTIVE: To observe the anti-convulsion effect of Shenpu Decoction (SPD). METHODS: Experiments were conducted in three acute convulsion models (cardiazol seizure, strychnine convulsion and maximal electrical shock). Models were divided into the control group (A, treated by normal saline), the high, middle and low dose SPD groups (B, C and D, treated with SPD 9 g/kg, 6 g/kg and 4 g/kg respectively), and the nitrazepam treated group (E). The anti-convulsion effect of SPD was evaluated by 50% convulsion dose (CD50) detected in each group. RESULTS: In cardiazol convulsion model, the CD50 detected in group A-E were 63.3 +/- 3.4 mg/kg, 116.2 +/- 3.4 mg/kg, 105.6 +/- 3.7 mg/kg, 74.0 +/- 3.7 mg/kg and 197.2 +/- 3.3 mg/kg respectively, while they were 0.71 +/- 0.04 mg/kg, 1.21 +/- 0.04 mg/kg, 1.19 +/- 0.04 mg/kg, 0.94 +/- 0.04 mg/kg and 1.16 +/- 0.04 mg/kg respectively in the strychnine convulsion model, and 67.1 +/- 2.6 V, > 140 V, 109.4 +/- 3.5 V, 84.4 +/- 3.1 V and 128.4 +/- 3.9 V in the maximal electrical shock model respectively. CONCLUSION: SPD has a good anticonvulsion effect.

Effects of heat processing and storage on flavanols and sensory qualities of green tea beverage.

This research was conducted to understand the effects of heat processing and storage on flavanols and sensory qualities of green tea extract. Fresh tea leaves were processed into steamed and roasted green teas by commercial methods and then extracted with hot water (80 degrees C) at 1:160 ratio (tea leaves/water by weight). Green tea extracts were heat processed at 121 degrees C for 1 min and then stored at 50 degrees C to accelerate chemical reactions. Changes in flavanol composition and sensory qualities of green tea extracts during processing and storage were measured. Eight major flavanols (catechin, epicatechin, gallocatechin, epigallocatechin, epicatechin gallate, catechin gallate, epigallocatechin gallate, and gallocatechin gallate) were identified in the processed tea extract. Among them, epigallocatechin gallate and epigallocatechin appeared to play the key role in the changes of sensory qualities of processed green tea beverage. The steamed tea leaves produced a more desirable quality of processed green tea beverage than the roasted ones.

Mutagenicity and aromatic amine content of fumes from heated cooking oils produced in Taiwan.

According to toxicological studies, there are several unidentified mutagens derived from cooking oil fumes appearing in kitchens of Chinese homes where women daily prepare food. Data are limited to an analysis of aromatic amines from cooking oil fumes, which are known to be carcinogenic for bladder cancer. Fume samples from three different commercial cooking oils frequently used in Taiwan were collected and analysed for mutagenicity in the Salmonella/microsome assay. Aromatic amines were extracted from the samples and identified by HPLC and confirmed by gas chromatography/mass spectrometry (GC/MS). Extracts from three cooking oil fumes were found to be mutagenic in the presence of S-9 mix. All samples contained 2-naphthylamine (2-NA) and 4-aminobiphenyl (4-ABP). Concentrations of 2-NA and 4-ABP were 31.5 and 35.7 microg/m3 in fumes from sunflower oil, 31.9 and 26.4 mg/m3 in vegetable oil, and 48.3 and 23.3 microg/m3 in refined-lard oil, respectively. Mutagenicities of the three cooking oil condensates were significantly reduced (P<0.05) by adding the antioxidant catechin (CAT) into the oils before heating. Significant difference existed between the amounts of aromatic amines with and without adding CAT (P<0.05). These results indicate that exposure to cooking oil fumes in Taiwan might be an important but controllable risk factor in the aetiology of bladder cancer.

Nitro-polycyclic aromatic hydrocarbon contents of fumes from heated cooking oils and prevention of mutagenicity by catechin.

According to earlier studies, fumes from cooking oils were found to be mutagenic and several polycyclic aromatic hydrocarbons (PAHs), (benzo(a)pyrene (B(a)P), benz(a)antracene (B(a)A), and dibenz(a,h)anthracene (DB(ah)A)) were identified. Fume samples from three different commercial cooking oils frequently used in Taiwan were collected and nitro-polycyclic aromatic hydrocarbons (NPAHs) were extracted from the samples and identified by HPLC chromatography. Extracts from three cooking oil fumes contained 1-nitropyrene (1-NP) and 1,3-dinitropyrene (1,3-DNP). Concentrations of 1-NP and 1,3-DNP were 1.1 +/- 0.1 and 0.9 +/- 0.1 micrograms/m3 in fumes from lard oil, 2.9 +/- 0.3 and 3.4 +/- 0.2 micrograms/m3 in soybean oil, 1.5 +/- 0.1 and 0.4 +/- 0.1 micrograms/m3 in peanut oil, respectively. The preventive effect of three natural antioxidants (gamma-tocopherol (TOC), lecithin (LEC), and catechin (CAT)) for the reduction of mutagenicity and amounts of PAHs and NPAHs of fumes from cooking oils were evaluated. Mutagenicity of cooking oil fumes occurred, and the concentration of B(a)P were significantly reduced (p < 0.05), by adding CAT into cooking oils before heating. B(a)A, DB(ah)A, and two NPAHs were not detected when the concentration of CAT was 500 ppm in all three cooking oil fumes. These results indicate that fumes of cooking oils contained PAHs and NPAHs that may be a risk factor for lung cancer among cooks and the carcinogens could be reduced by adding the natural antioxidant, catechin.

Mutagenicity and polycyclic aromatic hydrocarbon content of fumes from heated cooking oils produced in Taiwan.

According to epidemiologic studies, exposure of women to fumes from cooking oils appears to be an important risk factor for lung cancer. Fume samples from three different commercial cooking oils frequently used in Taiwan were collected and analyzed for mutagenicity in the Salmonella/microsome assay. Polycyclic aromatic hydrocarbons were extracted from the samples and identified by HPLC chromatography. Extracts from three cooking oil fumes were found to be mutagenic in the presence of S9 mix. All samples contained dibenz[a,h]anthracene (DB[a,h]A) and benz[a]anthracene (B[a]A). Concentration of DB[a,h]A and B[a]A were 1.9 and 2.2 micrograms/m3 in fumes from lard oil, 2.1 and 2.3 micrograms/m3 in soybean oil, 1.8 and 1.3 micrograms/m3 in peanut oil, respectively. Benzo[a]pyrene (B[a]P) was identified in fume samples of soybean and peanut oil, in concentrations of 19.6 and 18.3 micrograms/m3, in this order. These results provide experimental evidence and support the findings of epidemiologic observations, in which women exposed to the emitted fumes of cooking oils are at increased risk of contracting lung cancer.

Expression of the calpastatin gene segment during spermiogenesis in human testis: an in situ hybridization study.

Serum obtained from an infertile woman contained antibodies that agglutinate human sperm. The antibodies interacted with a sperm protein with an estimated M(r) of 17.5 kD. The cDNA coding the 17.5-kD protein was isolated from a human testis lambda gt11 expression library and identified as a segment of the calpastatin gene. Single-stranded 35S-labeled RNA probes were prepared from the calpastatin cDNA segment. Using the techniques of in situ hybridization, the calpastatin mRNA was located in spermatids of human testis. The results support a previous observation that the calpastatin segment is produced during spermiogenesis and suggest that transcription of the calpastatin gene occurred during the postmeiotic haploid stage of spermatogenesis.

Calpastatin gene in human testis.

Antibodies present in a serum obtained from an infertile woman interacted with a 17.5 kD glycoprotein (BS-17 component) extracted from human sperm by Western blot. Polyclonal antibodies were raised against the BS-17 component and used to identify positive staining clones from a human testis lambda gt11 expression library. The cDNA encoding the BS-17 component was isolated and its nucleotide sequence determined. The BS-17 cDNA contained 758 nucleotides with an open reading frame of 558 nucleotides encoding a polypeptide consisting of 186 amino acids. The BS-17 cDNA showed 99.7% homology in 758 nucleotides overlap with the 3' terminus of the gene coding calpastatin and 99.5% identity in 186 amino acid overlap with the carboxyl terminus of calpastatin. The BS-17 component of human sperm corresponds to the carboxyl terminus of calpastatin. This conclusion is supported by the finding that the polyclonal antibodies also interacted with a 84 kD protein corresponding to the M(r) of calpastatin.