[Review and perspective of clinical research involving chest tightness variant asthma in China].

Chest tightness variant asthma (CTVA) was first reported and named by Chinese scholars in 2013. It is a new clinical type of asthma characterized by chest tightness as the only or primary symptom, without typical asthma manifestations such as recurrent wheezing and shortness of breath, and without wheezing sounds heard during lung auscultation. The overall epidemiological data on CTVA is currently unavailable. Its pathogenesis is similar to that of typical asthma, involving eosinophilic airway inflammation. Due to the lack of typical clinical manifestations, insufficient knowledge of this disease in some clinicians and some other reasons, CTVA is susceptible to misdiagnosis or missed diagnosis. Currently, the diagnostic criteria for CTVA are: chest tightness as the only or primary symptom, without typical asthma symptoms and signs such as wheezing and shortness of breath, and with any one of the objective indicators of variable airflow limitation. Effective anti-asthma treatment is required, and other diseases that cause chest tightness, such as cardiovascular, digestive, nervous, muscular, and mental diseases should be excluded. CTVA treatment follows that of typical asthma, but the specific treatment duration is uncertain and may require long-term management. Traditional Chinese medicine has shown some therapeutic effects on CTVA. Most CTVA patients have a good prognosis after active anti-asthma treatment. This paper analyzes and summarizes the research of CTVA in China from 2013 and provides new perspectives for further exploration of CTVA.

[Comparative analysis of clinical diagnosis and treatment guidelines for drug-induced liver injury at home and abroad].

Drug-induced liver injury influencing factors are complex and have diverse clinical manifestations. Simple and reliable diagnostic methods are still deficient, and further classification of toxicological mechanisms is required. There are numerous pertinent discrepancies between domestic and international guidelines aimed at drug-induced liver injury diagnosis and treatment, with partial to no consensus on the content. The American Gastroenterological Association's 2021 Clinical Guidelines, the Asia-Pacific Association for the Study of the Liver's 2021 Consensus Guidelines, the Council for International Organizations of Medical Sciences' 2020 International Consensus, the European Society's Hepatology Committee's 2019 Clinical Practice Guidelines, and the 2015 Chinese Medical Association Guidelines are five influential clinical guidelines on drug-induced liver injury at home and abroad. The epidemiology, risk factors, diagnosis and evaluation, treatment management, and other contents, particularly traditional Chinese medicine, were compared and analyzed using other relevant consensus opinions or guidelines in order to improve understanding and provide a reference for clinical diagnosis and treatment of drug-induced liver injury.

A multicomponent screen for feeding behaviour and nutritional status in Drosophila to interrogate mammalian appetite-related genes.

OBJECTIVE: More than 300 genetic variants have been robustly associated with measures of human adiposity. Highly penetrant mutations causing human obesity do so largely by disrupting satiety pathways in the brain and increasing food intake. Most of the common obesity-predisposing variants are in, or near, genes expressed highly in the brain, but little is known of their function. Exploring the biology of these genes at scale in mammalian systems is challenging. We sought to establish and validate the use of a multicomponent screen for feeding behaviour phenotypes, taking advantage of the tractable model organism Drosophila melanogaster. METHODS: We validated a screen for feeding behaviour in Drosophila by comparing results after disrupting the expression of centrally expressed genes that influence energy balance in flies to those of 10 control genes. We then used this screen to explore the effects of disrupted expression of genes either a) implicated in energy homeostasis through human genome-wide association studies (GWAS) or b) expressed and nutritionally responsive in specific populations of hypothalamic neurons with a known role in feeding/fasting. RESULTS: Using data from the validation study to classify responses, we studied 53 Drosophila orthologues of genes implicated by human GWAS in body mass index and found that 15 significantly influenced feeding behaviour or energy homeostasis in the Drosophila screen. We then studied 50 Drosophila homologues of 47 murine genes reciprocally nutritionally regulated in POMC and agouti-related peptide neurons. Seven of these 50 genes were found by our screen to influence feeding behaviour in flies. CONCLUSION: We demonstrated the utility of Drosophila as a tractable model organism in a high-throughput genetic screen for food intake phenotypes. This simple, cost-efficient strategy is ideal for high-throughput interrogation of genes implicated in feeding behaviour and obesity in mammals and will facilitate the process of reaching a functional understanding of obesity pathogenesis.

[Observation of the therapeutic and characteristic effects of terlipressin on refractory cirrhotic ascites].

Objective: To observe the therapeutic effect of terlipressin on refractory ascites (RA) in cirrhosis, and its role and impact on acute kidney injury (AKI). Methods: A non-randomized controlled clinical trial data of 111 hospitalized cases of liver cirrhosis accompanied with RA was collected from Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Zhongshan Hospital of Hubei Province, The First Affiliated Hospital of Zhengzhou University, The First Affiliated Hospital of Medical School of Zhejiang University, and People's Hospital of Pudong New Area of Shanghai between March 2015 and March 2017. 26 cases of conventional treatment group (control group) were divided into two subgroups: RA without AKI (RA-NAKI) and RA with AKI (RA-AKI), and each subgroup consisted 13 cases. Patients with bacterial infection were treated with diuretics, albumin supplementation and antibiotics. 85 cases were presented in terlipressin combined treatment group, of which 27 cases were of RA-NAKI and 58 cases were of RA-AKI. Control group was injected terlipressin 1mg of intravenous drip or static push (once q6 h ~ 12 h) for more than 5 days. The treatment duration lasted for 2 weeks with 4 weeks of follow-up. Body weight, 24-hour urine volume, abdominal circumference, mean arterial pressure (MAP), liver and kidney function, anterior hepatic ascites, deepest point of ascites, and ultrasonographic detection of ascites in supine position before treatment, one and two weeks after treatment and 4 weeks after follow-up were compared. Count data were tested by χ (2). Samples of four groups at baseline were compared. One-way analysis of variance was used for normal distribution data and Kruskal-Wallis H test for non-normal distribution data. Repeated measures analysis of variance was used to compare the difference in efficacy between different time points before and after treatment in the group. The LSD method of one-way ANOVA was used to compare the two groups. A t-test of independent samples was used to compare the efficacy of different time series between the two groups. Mann-Whitney rank- sum test was used to compare the data of non-normal distribution between the two groups. Results: (1) Baseline data were compared among 4 subgroups of terlipressin RA-NAKI and control RA-AKI. Control group age was higher than that of terlipressin group, and the serum creatinine (SCr) of the RA-AKI group was higher than RA-NAKI group, and there was no significant difference in the rest of the baseline data and the combined medication (P > 0.05). (2) An intra-group comparison between control and trelipressin before and after treatment showed that all patients had lower body mass, abdominal circumference and deepest ascites, and higher serum albumin (P < 0.05). 24-hour urine volume and MAP was significantly increased in the terlipressin group, while the pre-ascites, SCr and child Turcotte Pugh (CTP) scores were decreased. Body weight, abdominal circumference, pre-ascites, and deepest ascites of the terlipressin group were decreased, while MAP was increased during the treatment and follow-up periods. The differences were statistically significant when compared with the control group at the same time (P < 0.05). There was a statistically significant difference in the increase of 24-h urine volume in the terlipressin group compared with the control group (P < 0.05). The decrease in SCr and CTP in the terlipressin group after 2 weeks of treatment and 4 weeks of follow-up was statistically significant compared with the control group (P < 0.05). (3) Among the two subgroups of RA-AKI and RA-NAKI in the terlipressin group, the baseline SCr value of the former was higher than that of the latter. After treatment, the body weight, abdominal circumference, pre-ascites, deepest ascites and CTP scores were decreased. In the RA-AKI group, 24-hour urine volume, MAP, SCr and serum albumin concentration were significantly increased. The difference between the two subgroups before and after treatment was compared, and the body weight of RA-AKI group at 1, 2 weeks of treatment and 4 weeks of follow-up was significantly lower than RA-NAKI group, which were (- 2.3 ± 0.2 vs. - 1.5 ± 0.2) kg, (- 4.1 ± 0.2 vs. - 2.6 ± 0.2) kg, (- 4.2 ± 0.3 vs. - 2.4 ± 0.3) kg, respectively. RA-NAKI group urine volume was significantly increased at 2 weeks of treatment and 4 weeks of follow-up, which was (468 ± 42 vs. 110 ± 131) ml, (272 ± 34 ml vs. 11 ± 112) ml, respectively. SCr reduction (18.3 ± 4.7 vs. 0.9 ± 2.4) µmol/l at 4 weeks of follow-up was apparent in RA-NAKI group, and the difference was statistically significant (P < 0.05). Conclusion: Addition of terlipressin to conventional treatment may significantly increase MAP, 24-h urine volume, improve renal function and promote ascites resolution in patients with refractory cirrhotic ascites. Moreover, its combination effect is more obvious in AKI patients, and adverse reactions are mild.

[Correlation of liver stiffness measured by FibroTouch and FibroScan with Ishak fibrosis score in patients with chronic hepatitis B].

Objective: To investigate the correlation of liver stiffness measured by FibroTouch (FT) and FibroScan (FS) with Ishak fibrosis score in patients with chronic hepatitis B. Methods: A total of 313 patients with chronic hepatitis B who visited Department of Liver Cirrhosis in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from November 2014 to May 2016 were enrolled. All the patients underwent liver biopsy, and FT and FS were used to determine liver stiffness measurement (LSM). Serum biochemical parameters were measured, and the aspartate aminotransferase-to-platelet ratio index (APRI) in a multi-parameter model of liver fibrosis and fibrosis-4 (FIB-4) index were calculated. The consistency between the results of four noninvasive examinations and Ishak fibrosis score was compared. The t-test was used for comparison of LSM determined by FT and FS. Pearson correlation analysis was used investigate the correlation between LSM determined by FT and FS; Spearman correlation analysis was used to investigate the correlation of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels and Knodell score with LSM determined by FT and FS; the correlation between LSM determined by FT and FS and fibrosis stage was analyzed by partial correlation analysis adjusted by Knodell score for liver inflammatory activity; Spearman correlation analysis was used for APRI, FIB-4, and fibrosis stage. Based on the Ishak fibrosis score, the receiver operating characteristic (ROC) curve was used to analyze the values of four noninvasive methods in the diagnosis of liver fibrosis. Results: There was no significant difference in LSM measured by FT and FS in all patients (15.75±9.42 kPa vs 15.42±10.52 kPa, P > 0.05) and Pearson correlation analysis indicated a significant positive correlation between them (r = 0.858, P < 0.01); serum ALT and AST levels and liver inflammatory activity were correlated with LSM determined by FT and FS. There was a significant positive correlation between LSM determined by FT and FS and fibrosis stage (r = 0.501 and 0.526, both P < 0.001), and APRI and FIB-4 were also positively correlated with fibrosis stage (r = 0.236 and 0.218, both P < 0.001). Based on the Ishak fibrosis score, in the diagnosis of fibrosis stages F3, F4, F5, and F6, the areas under the ROC curve were 0.915/0.856/0.839/0.816 for FT, 0.933/0.883/0.849/0.856 for FS, 0.618/0.630/0.608/0.638 for APRI, and 0.614/0.624/0.595/0.649 for FIB-4, and FT and FS had a significantly larger areas under the ROC curve than APRI and FIB-4. Conclusion: LSM determined by FT or FS has a good correlation with the Ishak fibrosis score, so FT and FS have a significantly better diagnostic performance for liver fibrosis than APRI and FIB-4.

[Aesthetic reconstruction strategy for postburn facial scar and its clinical effect].

OBJECTIVE: To explore the aesthetic reconstruction strategy for postburn facial scar and its clinical effect. METHODS: Three hundred and forty-two patients with postburn facial scars were hospitalized from January 2000 to December 2015. Local expanded flap or deltopectoral expanded flap was used for reconstruction according to the location and size of the facial scar. The forehead expanded flap could be chosen for the scar in dorsum nasi or inferior eyelid. The local expanded flap was chosen when the scar width was smaller than 5 cm in cheek, chin, and marginal mandible region. The expanded deltopectoral flap was chosen when the scar width was larger than 5 cm in cheek, chin, and marginal mandible region or the scar contracture was too serious to cause displacement of lips, nose, or eyelid, and the wound width was larger than 5 cm after release. The facial scars of 82 patients, with size ranged from 6.0 cm×2.5 cm to 15.0 cm×10.0 cm, were reconstructed with expanded local flaps. The facial scars of 260 patients, with size ranged from 8.0 cm×7.0 cm to 38.0 cm×13.0 cm, were reconstructed with expanded deltopectoral flaps. After expansion of 2 to 6 months, the facial scars were excised and completely released first of all. The transfer way of local flap and size of deltopectoral flap with pedicle were designed according to the size and shape of the wound. Three weeks after transfer of deltopectoral flap, flap delay procedure was conducted. One week later, the pedicle was severed from the flap to reconstruct the remaining scar. Anti-scar medicine, laser therapy, and elasticized fabric were used postoperatively on the scars in both donor and recipient sites. RESULTS: During the postoperative follow-up for 3 to 12 months, the flaps of 40 out of 82 cases reconstructed with expanded local flaps were in good color and texture. Before 2008, mild scar hyperplasia was observed in the incision of 19 patients; with application of laser after 2008, the number of patients with scar hyperplasia was decreased. During the postoperative follow-up for 3 to 12 months, the flaps of 90 out of 260 cases reconstructed with expanded deltopectoral flaps were in good color and texture. The expander was exposed from the incision in 15 patients, while it did not affect the later treatment. Nine unilateral flaps showed poor blood circulation at the distal end, and they were healed after dressing change. In the early phase, necrosis was observed in one flap after transfer, and it was healed after transplantation of free skin graft. Scar hyperplasia was observed in the chest donor site of one patient, and it was improved after laser therapy. CONCLUSIONS: Postburn facial scar could be reconstructed with local or deltopectoral flaps, following the principle of similarity. The expansion could increase the size of the flaps, reduce the thickness of the flaps, and lower the donor site damage.

A NOVEL OLEAGINOUS YEAST STRAIN WITH HIGH LIPID PRODUCTIVITY AND ITS APPLICATION TO ALTERNATIVE BIODIESEL PRODUCTION.

Five lipid-producing yeast strains, CHC08, CHC11, CHC28, CHC34, and CHC35, were revealed by Sudan Black B staining to contain lipid droplets within cells. Molecular analysis demonstrated that they were 2 strains of Candida parapsilosis, Pseudozyma parantarctica, Pichia manshurica, and Pichia occidentalis. Following batch fermentation, P. parantarctica CHC28 was found to have the highest biomass concentration, total lipids and lipid content levels. The major fatty acids in the lipids of this yeast strain were C16 and C18. Predictions of the properties of yeast biodiesel using linear equations resulted in values similar to biodiesel made from plant oils. Preliminary production of yeast biodiesel from P. parantarctica CHC28 was accomplished through esterification and transesterification reactions. It was found that yeast lipids with high acid value are easily converted to biodiesel at an approximately 90% yield. Therefore, it is possible to use crude lipids as alternative raw materials for biodiesel production.

Therapeutic efficacy of Traditional Chinese Medicine 319 recipe on hepatic fibrosis induced by carbon tetrachloride in rats.

BACKGROUND/AIMS: Hepatic fibrosis is a consequence of severe liver damage that occurs in many patients with chronic liver diseases. TCM 319 recipe is a Chinese Medicine formula which consists of six Chinese herbs. In this study, we investigated the anti-fibrotic efficacy and mechanisms of TCM 319 recipe. METHODS: Hepatic fibrosis in rats was induced by carbon tetrachloride (CCl4). 34 male adult SD rats were allocated into five groups (group 1-concomitant CCl4 and TCM 319 recipe for 8 weeks; group 2-CCl4 for 4 weeks and then CCl4 and TCM 319 recipe for 4 weeks; group 3-CCl4 alone for 8 weeks; group 4-TCM 319 recipe only for 8 weeks; group 5-untreated controls). After 8 weeks of treatment, serum ALT assay, liver tissue histological examination and immunostaining were carried out to examine the liver function and fibrosis degree. The expression levels of platelet derived growth factor (PDGF-B), PDGF-Rbeta, and transforming growth factor-beta 1 (TGF-beta1) were measured by quantitative RT-PCR and western blot. RESULTS: TCM 319 recipe reduced liver injury and attenuated hepatic fibrosis in group 1 compared with that in group 3. TCM 319 recipe suppressed the mRNA expression of tissue inhibitor of metalloproteinase 1 (TIMP-1). In addition, treatment with TCM 319 recipe significantly down-regulated mRNA expression of PDGF-B and PDGF-Rbeta, and it also suppressed protein expression of PDGF-Rbeta and TGF-beta1. CONCLUSIONS: TCM 319 recipe extracts could attenuate hepatic fibrosis induced by CCl4 in rats. The anti-fibrotic effect of TCM 319 recipe is associated with the down-regulation of mRNA expression of TIMP-1, PDGF-B and PDGF-Rbeta, and with the suppression of protein expression of PDGF-Rbeta and TGF-beta1.

Chromium supplementation enhances insulin signalling in skeletal muscle of obese KK/HlJ diabetic mice.

AIM: Chromium is an essential nutrient required for glucose and lipid metabolism. Laboratory and clinical evidences indicate that chromium supplementation may improve insulin sensitivity by enhancing intracellular signalling. Considerable evidence suggests that serine phosphorylation of insulin receptor substrate 1 (IRS1) at 307 residue (IRS1-Ser307) inhibits insulin signalling and results in peripheral insulin resistance. Therefore, we investigated whether chromium-associated insulin action was mediated by modulation of IRS1-Ser307 phosphorylation. METHODS: Male KK/HlJ mice (genetically obese and insulin resistant) were supplemented daily with chromium-containing milk powder or placebo for 7 weeks. In analysing functionally characterized insulin resistance, the changes of blood biochemicals, inflammatory factors and insulin signalling molecules in skeletal muscle were analysed. RESULTS: Using KK mice model, we demonstrated that daily supplementation of trivalent chromium-containing milk powder reduced serum levels of glucose, insulin and triglycerides, and improved glucose and insulin tolerance. Mechanistic study showed that chromium supplementation activated postreceptor insulin signalling such as increasing IRS1, IRS1 tyrosine phosphorylation, p85alpha regulatory subunit of phosphatidylinositol 3-kinase and glucose transporter 4 expression, stimulating Akt activity, downregulating c-Jun N-terminal kinase (JNK) activity and decreasing IRS1 ubiquitinization and insulin resistance-associated IRS1 phosphorylation (IRS1-Ser307) in skeletal muscle. In addition, chromium supplementation attenuated pro-inflammatory cytokine expression in both blood circulation and skeletal muscle. CONCLUSION: Our data suggest that chromium-containing milk powder supplementation can provide a beneficial effect in diabetic subjects by enhancing insulin signalling in skeletal muscle. The improvement in insulin signalling by chromium was associated with the decreased IRS1-Ser307 phosphorylation, JNK activity and pro-inflammatory cytokine production.

Frequency tuning and firing pattern properties of auditory thalamic neurons: an in vivo intracellular recording from the guinea pig.

We investigated the firing pattern and frequency tuning properties of medial geniculate body (MGB) neurons, through in vivo intracellular recordings in anesthetized guinea pigs. Twenty-two of the 25 physiological characterized neurons were anatomically identified. Ten neurons were located in the ventral division of the medial geniculate body (MGv) (seven in pars ovoidea (OV) and three in the pars lateralis (LV)). Eight were located in the dorsal division (MGd), and four in the medial division (MGm). OV neurons showed a uniform, phasic ON response with high frequency selectivity. Functionally, they are interpreted as relaying spectral information with high reliability. LV neurons exhibited various patterns: phasic, tonic and excitatory postsynaptic potentials (EPSP) with a spike train. These high magnitude EPSPs are proposed to convey temporal information of the auditory signals with more encoding power. MGd neurons had relatively low best frequencies while MGm neurons had high intensity threshold, broader frequency selectivity, and a tonic response pattern. Tonic firing is likely to impose a strong impact onto wide cortical area and amygdala. When hyperpolarized with current injection, MGB neurons evoked low-threshold calcium spikes. Distinct change in these spike numbers was observed among MGv and MGd neurons as compared with MGm neurons, implying their differential roles. MGm neurons are more modulatory in nature, while the long lasting bursts of low-threshold calcium spikes observed in MGv and MGd neurons probably participate in propagating the sleep oscillations.

Repellent and insecticidal activities of essential oils from Artemisia princeps and Cinnamomum camphora and their effect on seed germination of wheat and broad bean.

Repellent and insecticidal activities of essential oils extracted from leaves of Artemisia princeps Pamp and seeds of Cinnamomum camphora (L.) Presl. against storage pests Sitophillus oryzae L. and Bruchus rugimanus Bohem were investigated. Results showed that the two individual oils displayed good, but their mixture (1:1) exhibited much better repellent activities at concentrations from 250 to 1000 microg g(-1) and insecticidal actions at concentrations 1000 microg g(-1) against the test beetles S. oryzae and B. rugimanus. Oils from A. princeps and C. camphora applied individually were significantly toxic to seed germination of wheat at 500 microg ml(-1). However, no toxic effects were found when the two oils were mixed (1:1 w/w) at the same concentration. These observations indicated that the mixture of the two plant-derived oils had a synergic effect and could be used in the control of storage pests.

Absolute configuration of keisslone, a new antimicrobial metabolite from Keissleriella sp. YS4108, a marine filamentous fungus.

In order to purify enough material for establishing the absolute stereochemistry of the new antifungal metabolite 3,6,8-trihydroxy-3-[3,5-dimethyl-2-oxo-3(E)-heptenyl]-2,3-dihydronaphthalen-1(4H)-one produced by Keissleriella sp., a marine filamentous fungus (strain number: YS 4108), a repeated growth and fractionation of the fungal culture was performed to give instead a new antimicrobial metabolite, keisslone (1), the structure of which was elucidated on the basis of spectral analyses including homo- and hetero-nuclear correlation NMR experiments (HMQC, COSY, NOESY and HMBC). The absolute configuration of metabolite 1 was determined mainly by its CD data and NOESY spectrum. The compound 1 was shown to be inhibitory against the human pathogenic fungi Candida albicans, Trichophyton rubrum and Aspergillus niger with MICs of 50, 70, 40 microg/mL, respectively. In order to purify enough material for establishing the absolute stereochemistry of the new antifungal metabolite 3,6,8-trihydroxy-3-[3,5-dimethyl-2-oxo-3(E)-heptenyl]-2,3-dihydronaphthalen-1(4 H)-one produced by Keissleriella sp., a marine filamentous fungus (strain number: YS 4108), a repeated growth and fractionation of the fungal culture was performed to give instead a new antimicrobial metabolite, keisslone (1), the structure of which was elucidated on the basis of spectral analyses including homo- and hetero-nuclear correlation NMR experiments (HMQC, COSY, NOESY and HMBC). The absolute configuration of metabolite 1 was determined mainly by its CD data and NOESY spectrum. The compound 1 was shown to be inhibitory against the human pathogenic fungi Candida albicans, Trichophyton rubrum and Aspergillus niger with MICs of 50, 70, 40 microg/mL, respectively.

Antifungal metabolite with a new carbon skeleton from Keissleriella sp. YS4108, a marine filamentous fungus.

In addition to four known metabolites (4-acetyl-6,8-dihydroxy-5-methylisocoumarin, 6,8-dihydroxy-3-methylisocoumarin, 6,8-dihydroxy-3,5,7-trimethylisocoumarin and 3,3'-oxy-(5-methyl)-phenol), bioassay-guided fractionation of the culture of Keissleriella sp., a marine filamentous fungus (strain number: YS 4108), afforded an antifungal metabolite with a new carbon skeleton whose structure was elucidated spectrometrically as 3,6,8-trihydroxy-3-[3,5-dimethyl-2-oxo-3(E)-heptenyl]-2,3-dihydronaphthalen-1(4H)-one. In vitro antifungal assays of all isolates revealed that the new metabolite and 3,3'-oxybis[5-methylphenol] were inhibitory to the growth of the human pathogenic fungi Candida albicans, Tricophyton rubrum and Aspergillus niger with MICs of the former being 40, 20 and 80 microg/ml, and those of the latter 10, 30 and 50 microg/ml, respectively.

Antifungal activity of Artemisia annua endophyte cultures against phytopathogenic fungi.

Artemisia annua, well recognized for its production of antimalarial drug artemisinin, is seldom attacked by any of phytopathogenic fungi, which could be partially associated with the presence of endophytes. Present investigation is aiming at disclosing whether the endophytes inside A. annua produce antifungal substances. A total of 39 endophytes were isolated and fermented, and the ferment broth was evaluated in vitro for the antifungal activity against crop-threatening fungi Gaeumannomyces graminis var. tritici, Rhizoctonia cerealis, Helminthosporium sativum, Fusarium graminearum, Gerlachia nivalis and Phytophthora capsici. These plant pathogens are still causing wheat take-all, sharp eyespot, common rot, scab, snow mould, and pepper phytophthora blight, respectively. Out of 39 endophytes investigated, 21 can produce in vitro substances that are inhibitory to all or a few of the tested phytopathogens whereas the rest yielded nothing active. Moreover, the most active broth of endophyte IV403 was extracted with EtOAc and n-butanol, and comparisons of the antifungal activity of the extracts indicated that the major active metabolites were EtOAc-extractable.

Factorial designs combined with the steepest ascent method to optimize serum-free media for CHO cells.

A serum free medium for recombinant CHO NTHU 108 cell growth and fusion protein (CD20 linked to a human IgG-Fc gamma4 fragment) synthesis were systematically developed using factorial designs combined with the steepest ascent method. Experimental results indicate that the optimal composition of serum replacement for specific fusion protein production was 1% SITE (selenium, insulin, transferrin, ethanolamine), 0.3 g/L yeast extract, and 0.09% linoleic acid-BSA. Cell growth and fusion protein production of the adapted CHO NTHU 108 cultured in Iscove's modified Dulbecco's medium supplemented with these serum substitutes were comparable to those in the Ex-Cell 301 commercial serum-free medium. These serum substitutes can also promote CHO cell growth and fusion protein production in nine kinds of commercial media. The low protein content of the developed medium facilitates downstream processing and product purification.

Nitric oxide synthase inhibitor ameliorates oral total parenteral nutrition-induced barrier dysfunction.

The expression of inducible nitric oxide synthase (iNOS) is increased in the intestine and results in mucosal damage after endotoxin challenge. Although the oral administration of total parenteral nutrition (TPN) solution promotes bacterial translocation (BT) and increases the intestinal permeability, the role of NO in the nutrition-induced loss of mucosal barrier function remains unclear. The distribution of fluorescein isothiocyanate-dextran (FITC-dextran, 4400) across the lumen of small intestine in rat was examined to investigate the role of NOS activity on the intestinal permeability under oral TPN feeding. Fifty-one rats were randomly divided into 4 groups. Group I (control group) was fed with rat chow, group II received TPN solution orally. Groups III and IV received TPN solution supplemented with NOS inhibitors. On day 9, FITC-dextran was injected into the intestinal lumen. After 30 min, blood samples were taken from portal vein and analyzed for plasma FITC-dextran level by fluorescence spectrophotometry. Samples of small intestine were frozen and sectioned in a cryostat for morphological and NOS histochemical studies. Homogenates of small intestine were used for NOS activity measurement. The plasma level of FITC-dextran showed a significant increase (P < 0.05) in rats fed with oral TPN compared with the control ones. Supplement with NOS inhibitors significantly decreased the intestinal permeability in groups III and IV compared with group II. Similarly, the total NOS activities showed a significant 2-fold increase (P< 0.05) in group II, and NOS inhibitors decreased the elevated NOS activity. These data suggest that oral TPN feeding for 9 days leads to an increase in permeability to dextran and the total NOS activity of small intestine, and both induction of the intestinal permeability and NOS activity were inhibited by treatment with NOS inhibitors. Addition of S-methylisothiourea (SMT), an iNOS selective inhibitor, profoundly inhibited 66% of the induced iNOS activity (P < 0.05) and reduced 74% of the diet-induced increase in intestinal permeability (P < 0.05) in group II. The induced permeability change in rats receiving oral TPN is mainly due to the activity of intestinal mucosal iNOS. The induction of iNOS is an important mediator for intestinal barrier dysfunction. Administration of SMT, which specifically decreases iNOS activity, is useful in the prevention of diet-induced barrier failure.

[Studies on quality standards for Pollen Typhae(puhuang)].

OBJECTIVE: To work out quality standards for Pollen Typhae(Puhuang). METHOD: Isorhamnetin-3-O-neohesperidoside, typhaneoside, isorhamnetin and beta-sitosterol in Pollen Typhae marketed in ten cities of China were identified by TLC. The contents of isorhamnetin-3-O-neohesperidoside and typhaneoside were determined by HPLC. RESULT: The method is sensitive and specific for identifying isorhamnetin-3-O-neohesperidoside, typhaneoside, isorhamnetin and beta-sitosterol. The linear ranges of isorhamnetin-3-O-neohesperidoside and typhaneoside were 0.188-0.940 microgram with and average recovery of 97.77%; and 0.164-0.820 microgram with an average recovery of 98.42% respectively. CONCLUSION: The method can be used both qualitatively and quantitatively as standards for the quality control of Pollen Typhae.

Enterovesical fistula: experiences with 41 cases in 12 years.

BACKGROUND: A retrospective analysis of enterovesical fistula treated at Chang Gung Memorial Hospital was conducted to determine the optimal diagnosis and management of this disease. METHODS: The records of 41 patients who presented from 1984 to 1996 and had a final diagnosis of enterovesical fistula were retrospectively reviewed. The etiology, symptoms on presentation, diagnostic tools, and modality of treatment were analyzed. RESULTS: The majority of these cases were associated with malignancy (38, 92.7%), and the others with diverticulitis (2, 4.9%) and iatrogenic causes (1, 2.4%). In those with malignancy, 15 patients (39.5%) were found to have tumor recurrence. The most frequent symptom in enterovesical fistula was fecaluria (58.5%), followed by abdominal pain (22%) and dysuria (14.6%). Diagnostic tools included the barium enema, cystography, and cystoscopy; these methods could identify the fistula in 63.2%, 60%, and 53.8% of the patients, respectively. Methods of management included diversion only (39%), one-stage fistula repair (36.6%), and watchful surveillance (24.4%). CONCLUSION: Enterovesical fistula should be considered if fecaluria, pneumaturia, or persistent non-specific urinary tract infection present as the initial complaint. A thorough surgery for a possible underlying malignancy is mandatory when confronted with enterovesical fistula, since the incidence of inflammatory bowel disease is low in this area. An abdominal computer tomography (CT) scan, barium enema, and cystogram can be useful diagnostic tools. Treatment of this entity should be individualized according to each patients clinical status.

Effect of supplementary antioxidant vitamin intake on carotid arterial wall intima-media thickness in a controlled clinical trial of cholesterol lowering.

BACKGROUND: There is accumulating experimental, epidemiological, and clinical evidence of an association between anti-oxidant vitamin intake and reduced risk of coronary heart disease. Using data from the Cholesterol Lowering Atherosclerosis Study (CLAS), we explored the association of self-selected supplementary antioxidant vitamin intake on the rate of progression of early preintrusive atherosclerosis. METHODS AND RESULTS: CLAS was an arterial imaging trial in which nonsmoking 40- to 59-year-old men with previous coronary artery bypass graft surgery were randomized to colestipol/niacin plus diet or placebo plus diet. The rate of progression of early preintrusive atherosclerosis was determined in 146 subjects using high-resolution B-mode ultrasound quantification of the distal common carotid artery far wall intima-media thickness (IMT). From the nutritional supplement database, 22 subjects had an on-trial average supplementary vitamin E intake of > or = 100 IU per day (high users) and 29 subjects had an average on-trial supplementary vitamin C intake of > or = 250 mg per day (high users). Within the placebo group, less carotid IMT progression was found for high supplementary vitamin E users when compared with low vitamin E users (0.008 versus 0.023 mm/y, P = .03). No effect of vitamin E within the drug group was found. No effect of vitamin C within the drug or placebo group was found. CONCLUSIONS: Supplementary vitamin E intake appears to be effective in reducing the progression of atherosclerosis in subjects not treated with lipid-lowering drugs while the process is still confined to the arterial wall (early preintrusive atherosclerosis).