呋喃那斯对暴发性鱼病的防治方法研究

呋喃那斯对暴发性鱼病的ＢＳＫ－１０菌株的最低抑制浓度为０．０１９ｍｇ／Ｌ,比呋喃唑酮的用量低３２倍。用０．２和０．５ｍｇ／Ｌ浸泡药浴病鱼的治愈率为９０％和１００％;５０ｍｇ／Ｌ呋喃唑酮的治愈率为０。１０ｍｇ／Ｌ和２０ｍｇ／Ｌ药浴１０分种的死鱼时间比５０ｍｇ／Ｌ呋喃唑酮迟１天。以０．５和４．０μｇ／ｇ腹腔注射给药的防治效果也优于同剂量的呋喃唑酮。对鲢以１００ｍｇ／Ｌ药浴２小时或白鲫ｉｐ６００ｍｇ／ｋ     

牛微量白细胞样品的处理及其Y染色体特异DNA的扩增

本文建立了牛微量白细胞样品的处理及其Ｙ染色体特异ＤＮＡ的扩增的方法。采集成年公牛全血,用ＥＤＴＡ抗凝血。分离白细胞,用０．１４５ｍｏｌ／ＬＮａＣｌ洗净,用０．１４５ｍｏｌ／ＬＮａＣｌ稀释,计数。分别将０．５、５０、５００细胞在含１０ｍｍｏｌ／ＬＴｒｉｓ－ＨＣｌ、５０ｍｍｏｌ／ＬＫＣｌ、２ｍｍｏｌ／ＬＭｇＣｌ２、０．４５％ＮＰ－４０、０．４５％Ｔｗｅｅｎ－２０、０．１ｍｇ／ｍＬ蛋白酶Ｋ、总体积２０μ     

胡杨离体器官发生及试管无性系的建立

研究了离体条件下胡杨（Ｐｏｐｕｌｕｓ　ｅｕｐｈｒａｔｉｃａ　Ｏｌｉｖｅｒ）茎段、叶片及愈伤组织的器官发生和植株再生技术。离体培养以ＭＳ为基本培养基并附加４０ｍｇ／Ｌ腺嘌呤和５００ｍｇ／Ｌ水解乳蛋白。离体叶片和茎段在ＢＡ为０．５ｍｇ／Ｌ和ＮＡＡ为０．５ｍｇ／Ｌ的培养基上诱导产生愈伤组织,并在含０．２５ｍｇ／ＬＢＡ和０．５ｍｇ／ＬＮＡＡ的培养基上继代增殖。ＢＡ为０．５ｍｇ／Ｌ和ＮＡＡ为０．１ｍｇ／Ｌ可诱导叶片和愈伤组织发生不定芽,诱导频率分别为１００％和８２．９％,对于茎段,ＢＡ和ＮＡＡ分别为０．１ｍｇ／Ｌ和０．０１ｍｇ／Ｌ时诱导不定芽频率可达８３％。试管苗在大量元素减半并附加０．０１５ｍｇ／ＬＮＡＡ的ＭＳ培养基上诱导生根,生根率达８６．２％。     

肠通透性改变的研究初报

本文用气相色谱测定烧伤猪和羊的ＭＳＯＦ动物模型尿内乳果糖和甘露醇分泌比率（Ｌ／Ｍ）。烧伤猪治疗组７２小时Ｌ／Ｍ分泌比率为０．０２４±０．００７,而对照组为０．０２６±０．００３,伤前Ｌ／Ｍ分泌比率为０．００７±０．００５,与伤前比较,无显著性差异。这说明Ｌ／Ｍ分泌比率升高与烧伤前期治疗及伤后天数无关。而在羊的ＭＳＯＦ的动物模型内Ｍ组中,输入内毒素４小时及伤后３天的Ｌ／Ｍ分泌比率为０．６２±０．２、１．７０±０．６０,对照组为０．２８±０．２５、０．２０±０．１０,伤前为０．０２９±０．０２,Ｌ／Ｍ分泌比率升高显著（Ｐ＜０．０５）,提示内毒素血症可能与肠通透性改变之间具有密切联系。     

中麻黄悬浮培养体系的建立

本文用中麻黄无菌苗为外植体,其切段培养在附加２ｍｇ／Ｌ２,４－Ｄ和０．５ｍｇ／Ｌ　６　ＢＡ的ＭＳ培养基上,全部脱分化形成白色疏松愈伤组织。愈伤组织继代培养于ＭＳ＋０．５ｍｇ／Ｌ２,４－Ｄ＋０．２ｍｇ／Ｌ６ＢＡ＋０．２ｍｇ／Ｌ　ＮＡＡ＋４％蔗糖的培养某上。以继代培养愈伤组织为材料进行悬浮培养,培养基为附加０．２ｍｇ／Ｌ２,４－Ｄ＋０．１ｍｇ／Ｌ６ＢＡ＋０．１ｍｇ／ＬＮＡＡ＋２％蔗糖的ＭＳ液体培养基,得到分散性好,细胞形状接近圆形,细胞大小均一,细胞团多由２－３０个细胞组成的悬浮培养体系。第三代悬浮培养细胞增长率为０．３５ｇ·ｆｗ／２０ｍｌ·ｄ,细胞有丝分裂指数为１１．２％。条件培养和高密度接种可缩短延迟期,条件培养不能提高分裂指数,１ｇ／１０ｍｌ接种密度可使分裂指数提高至２１．２％。     

气相色谱法测定人白蛋白制品中辛酸钠含量

本文报导用气相色谱法测定人白蛋白制品中辛酸钠含量。样品用正庚酸作内标,经酸化、氯仿抽提、浓缩后,通过ＨＰ－ＩＮＮＯＷａｘ柱,以氢火焰检测器测定。本法色谱峰形好,平均回收率及变异系数分别为９８．５４％、１．３９％。测定线性范围在１０７μｇ／０．２５ｍｌ～７２０μｇ／０．２５ｍｌ。最小检测限为７．４１０μｇ。实验操作简便,样品及溶剂用量少,可作为人白蛋白制品中辛酸钠含量的检测方法。     

洋桔梗F1无菌播种和试管苗的快速繁殖

洋桔梗Ｆ１种子在１／２ＭＳ培养基中发芽,其种子苗的茎尖和带叶茎段,在含０．１ｍｇ／Ｌ６－ＢＡ,０．０３ｍｇ／Ｌ ＮＡＡ的３／４培养基中,可连续进行继代培养,增殖大量从生苗,无根苗扦插在珍珠岩基质中约２０天发根,成活率达９５％。     

绿色功夫和锐劲特对食用菌害虫的防治效果测定

室内毒力测定和菇房药效试验表明,绿色功夫和锐劲特对食用菌害虫眼蕈蚊和果蝇有较强的毒力和较好的防治效果。绿色功夫对眼蕈蚊和果蝇的ＬＣ５０分别为５．０８ｍｇ／Ｌ和２．４５ｍｇ／Ｌ。锐劲竺对眼蕈蚊和果蝇的ＬＣ５０分别为８．７５ｍｇ／Ｌ和８．５０ｍｇ／Ｌ。菇房防治效果显示：用质量浓度１２．５ｍｇ／Ｌ和６．２５ｍｇ／Ｌ的绿色功夫液浇淋和务菌代防治效果达８８．６％、８１．８％和７５．９％、６５．１％。用质量浓     

向日葵下胚轴原生质体高频率愈伤组织形成与根发生

向日葵籽苗下胚轴原生质体,培养在含有ＢＡ０．５ｍｇ／Ｌ,２,４－Ｄ０．５ｍｇ／Ｌ,ＮＡＡ０．１ｍｇ／Ｌ和葡萄糖０．５５ｍｇ／Ｌ的改良Ｋａｏ培养基中,２４～２８ｈ后,原生质体开始分裂。包埋在琼脂糖０．６％中的原生质体,培养５ｄ后,分裂频率达９５％以上。生长旺盛的小愈伤组织转移到含有２ｉｐ０．１ｍｇ／Ｌ,ＩＡＡ０．０１ｍｇ／Ｌ,腺嘌呤４０ｍｇ／Ｌ和ＧＡ３０．０１ｍｇ／Ｌ的Ｔｈｏｍｐｓｏｎ液体培养基上１３ｄ后,原生质体诱导的少数愈伤组织发生根分化。     

玉米雌穗离体培养诱导孤雌生殖结实

离体条件下,用１６种不同的玉米材料未授粉的雌穗进行整个直插或切段直插培养。培养在２８℃培养箱和室内散射光下,其培养基为：ＭＳ＋０．１ｍｇ／Ｌ２,４－Ｄ＋２ｍｇ／ＬＮＡＡ＋２ｍｇ／ＬＫＴ＋５００ｍｇ／ＬＬＨ＋４％蔗糖＋０．６％琼脂（ＰＨ５．８）和ＭＳ＋０．０５ｍｇ／Ｌ２,４－Ｄ＋２ｍｇ／ＬＮＡＡ＋２ｍｇ／ＬＫＴ＋４％蔗糖＋０．６％琼脂（ＰＨ５．８）。从未授粉的雌穗（成熟胚囊期）获得结实。这些种子萌发     

Resolution of common dietary sugars from probe sugars for test of intestinal permeability using capillary column gas chromatography

BACKGROUND: The most widely accepted method for the evaluation of intestinal barrier integrity is the measurement of the permeation of sugar probes following an oral test dose of sugars. The most-widely used sugar probes are sucrose, lactulose, mannitol and sucralose. Measuring these sugars using a sensitive gas chromatographic (GC) method, we noticed interference on the area of the lactulose and mannitol peaks. METHODS: We tested different sugars to detect the possible makeup of these interferences and finally detected that the lactose interferes with lactulose peak and fructose interferes with mannitol peak. On further developing of our method, we were able to reasonably separate these peaks using different columns and condition for our assay. Sample preparation was rapid and simple and included adding internal standard sugars, derivitization and silylation. We used two chromatographic methods. In the first method we used Megabore column and had a run time of 34 min. This resulted in partial separation of the peaks. In the second method we used thin capillary column and was able to reasonably separate the lactose and lactulose peaks and the mannitol and fructose peaks with run time of 22 min. RESULTS: The sugar probes including mannitol, sucrose, lactulose, sucralose, fructose and lactose were detected precisely, without interference. The assay was linear between lactulose concentrations of 0.5 and 40 g/L (r(2)=1.000, P<0.0001) and mannitol concentrations of 0.01 and 40 g/L (r(2)=1.000). The sensitivity of this method remained high using new column and assay condition. The minimum detectable concentration calculated for both methods was 0.5 mg/L for lactulose and 1 mg/L for mannitol. CONCLUSION: This is the first report of interference of commonly used sugars with test of intestinal permeability. These sugars are found in most of fruits and dairy products and could easily interfere with the result of permeability tests. Our new GC assay of urine sugar probes permits the simultaneous quantitation of sucralose, sucrose, mannitol and lactulose, without interference with lactose and fructose. This assay is a rapid, simple, sensitive and reproducible method to accurately measure intestinal permeability.     

Gas chromatography applied to the lactulose—mannitol intestinal permeability test

Intestinal permeability can be modified by various illnesses, trauma and sepsis. Alterations of the intestinal wall can facilitate the diffusion of potentially harmful substances such as endotoxins, as well as bacterial translocation. We describe the validation of a capillary gas chromatographic method for the determination of mannitol and lactulose, used as intestinal permeability probes. The method is linear up to 3 g/l for mannitol and 300 mg/l for lactulose; recovery from overload samples is between 92 to 110%. Intra-assay coefficients of variation (C.V.s) were 2.7 and 6.8% for mannitol and lactulose, respectively, and inter-assay C.V.s were 8.9 and 9.3%. Normal values for 25 healthy subjects (mean ± S.D.) were 14.5 ± 3.1% and 0.27 ± 0.15% for mannitol and lactulose, respectively. The GC method presented is rapid and precise.     

Studies on intestinal permeability of cirrhotic patients by analysis lactulose and mannitol in urine with HPLC/RID/MS

The method for separation and determination of lactulose (L) and mannitol (M) in urine was developed by HPLC with a refractive index detector (RID). The linearity ranged from 5 to 1000 microg/mL for L and M, respectively. Recoveries ranged from 93.1% to 97.1%. The intra- and inter-day relative standard deviations of peak area were between 0.8-1.4% (n=3) and 1.4-3.6% (n=3). The limits of detection were obtained with 1.40 microg/mL for L and 1.65 microg/mL for M. The ratios of L/M in the urine samples for the spontaneous ascitic fluid infection (SAI), sterile ascitic fluid (SA) patients, and healthy volunteers (HV) were determined. The results showed well the correlations among the L/M ratio, intestinal permeability (IP) and the illness status of patients, and also indicated lactulose could improve the IP of SAI patients. The peaks of L and M in chromatograms were identified by electrospray ionization/mass spectrometry (ESI/MS), which ensured the accurate measurement of the ratio L/M. This method presented a rapid, accurate, and practical technique for determining IP in clinical practice and investigating the pathology of hepatocirrhosis.     

Gas chromatographic method for detection of urinary sucralose: application to the assessment of intestinal permeability

We developed a capillary column gas chromatography (CCGC) method for the measurement of urinary sucralose (S) and three other sugar probes including, sucrose, lactulose (L) and mannitol (M) for use in in vivo studies of intestinal permeability. We compared the capillary method with a packed column gas chromatography (PCGC) method. We also investigated a possible role for sucralose as a probe for the measurement of whole gut permeability. Sample preparation was rapid and simple. The above four sugars were detected precisely, without interference. We measured intestinal permeability using 5- and 24-h urine collections in 14 healthy volunteers. The metabolism of sugars was evaluated by incubating the intestinal bacteria with an iso-osmolar mixture of mannitol, lactulose and sucralose at 37 degrees C for 19 h. Sugar concentrations and the pH of the mixture were monitored. The use of the CCGC method improved the detection of sucralose as compared to PCGC. The average coefficient of variation decreased from 15% to 4%. It also increased the sensitivity of detection by 200-2000-fold. The GC assay was linear between sucralose concentrations of 0.2 and 40 g/l (r=1.000). Intestinal bacteria metabolized lactulose and acidified the media but did not metabolize sucralose or mannitol. The new method for the measurement of urinary sucralose permits the simultaneous quantitation of sucrose, mannitol and lactulose, and is rapid, simple, sensitive, accurate and reproducible. Because neither S nor M is metabolized by intestinal bacteria, and because only a tiny fraction of either sugar is absorbed, this pair of sugar probes appears to be available for absorption throughout the GI tract. Thus, the 24-h urinary concentrations of S and M, or the urinary S/M ratio following an oral dose of a sugar mixture, might be good markers for whole gut permeability.     

Lactulose and mannitol intestinal permeability detected by capillary electrophoresis

Aim of this study was to set up a method by capillary electrophoresis to detect lactulose and mannitol in urine after an oral load, and to estimate the intestinal permeability in controls and in type I diabetes patients. The underivatized carbohydrates were monitored by indirect UV detection using sorbate, cetyltrimethylammonium bromide and LiOH as background electrolyte. Urines were purified by solid phase extraction, shaken with cation exchange resin, filtered and analysed. Carbohydrates migrated in <10 min in relation to their pK(a) and M(r). Controls (n = 33) and patients (n = 23) had an excretion ratio lactulose/mannitol 0.025 (0.018-0.051) and 0.067 (0.050-0.127), respectively (p < 0.01, median, interquartile range).     

Intestinal permeability in children with Crohn's disease and coeliac disease.

Mannitol and lactulose were used as probe molecules to measure intestinal permeability in children with active small-bowel Crohn''s disease and with untreated coeliac disease. Mannitol and lactulose were administered by mouth in a moderately hypertonic solution (580 mmol (mosmol)/l), and results were expressed as the ratio of the molecules excreted in urine over five hours. Patients with Crohn''s disease had a sixfold increase in permeability (due to increased lactulose permeability) and those with coeliac disease a fivefold increase (due to decreased mannitol permeability). From these results the test offers potential as a noninvasive investigation in children with small-bowel disease.     

Intestinal permeability to lactulose and mannitol in growing rats with iron-deficiency anemia

Iron deficiency can have nonhematological manifestations, some of which may affect the gastrointestinal tract. The aim of this study was to determine if iron-deficiency anemia in growing rats affected small-bowel permeability as assessed by the urinary ratio of lactulose and mannitol. Thirty-seven male Harlan Sprague-Dawley rats (21 d of age) were randomly divided into two groups and fed either an iron-deficient (n=19) or an iron-sufficient diet (n=18) that contained either 13.5 or 43.8 mg of iron/kg diet, respectively. Animals were evaluated between 25 and 38 d of dietary treatment. Intestinal permeability was assessed by measuring the lactulose/mannitol urinary ratio following administration of a solution that contained the two sugars. At the end of the study, the mean body weight of rats fed the low-iron diet was approx 95% that of the controls. The mean hemoglobin (g/dL) was significantly lower in the low-iron diet group (11.2±1.4) than in the control group (16.9±0.8) (p=0.001). The liver iron concentration (μg/g) of the anemic group (41.4±4.7) was also statistically (p=0.001) lower than in the control group (116.6±18.2). The lactulose/mannitol ratio was lower in the anemic rats (2.0±0.7) than in the control group (2.6±0.7) (p=0.008), a finding that is not suggestive of intestinal mucosal atrophy, previously described in anemic children.     

双歧杆菌四联活菌片联合乳果糖对乙肝肝硬化患者肠道菌群 和肠黏膜屏障功能及肝功能水平的影响

目的研究双歧杆菌四联活菌片联合乳果糖对乙肝肝硬化患者肠道菌群、肠黏膜屏障功能及肝功能水平的影响。方法选择2017年12月至2018年12月于我院感染科住院治疗的乙肝肝硬化患者80例,按照随机数字表法分为试验组和对照组各40例。两组患者常规予以抗病毒及保肝治疗。试验组患者加用双歧杆菌四联活菌片和乳果糖口服液。对照组患者仅给予乳果糖口服液。检测患者肠道肠杆菌、肠球菌、双歧杆菌、乳杆菌和白假丝酵母数量。测定患者血清内毒素和血清二胺氧化酶水平及尿乳果糖/甘露醇(L/M)比值。记录两组患者丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)和总胆红素(TBIL)水平。观察治疗过程中的药物不良反应发生情况。结果治疗4周后,试验组患者肠道肠球菌数量高于治疗1周后(18.41±2.92 vs 18.32±3.06),同时试验组患者肠道双歧杆菌数量与治疗前、治疗1周后以及对照组同时期相比差异也有统计学意义(均P0.05)。试验组患者治疗后肠道乳杆菌和白假丝酵母水平与治疗前相比差异均有统计学意义(均P0.05)。治疗4周后,对照组患者肠道肠杆菌水平高于治疗1周后(F=10.192,P=0.019);而白假丝酵母水平高于治疗前(F=8.567,P=0.024)。治疗前两组患者血清内毒素、血清二胺氧化酶和尿L/M以及AST、ALT、TBIL水平差异无统计学意义(均P0.05)。治疗1个月后,两组患者血清内毒素、血清二胺氧化酶和尿L/M以及AST、ALT、TBIL均较治疗前明显下降(均P0.05),且试验组的下降幅度较对照组更大(均P0.05)。两组患者均未发生不良反应。结论双歧杆菌四联活菌片联合乳果糖可以改善乙肝肝硬化患者肝功能状态,调节失衡的肠道菌群,降低肠黏膜通透性,值得临床推广。     

Cellobiose and lactulose coupled with mannitol and determined using ion-exchange chromatography with pulsed amperometric detection,are reliable probes for investigation of intestinal permeability

Lactulose/mannitol and cellobiose/mannitol tests are currently used in the investigation of intestinal permeability (IP) in many gastrointestinal diseases. The aim of this study was to produce a good technique for the determination and comparison of the above-mentioned sugar probes to overcome the problem caused by the presence of significant glycosuria in patients affected by particular metabolic disorders such as diabetes mellitus. Tests were performed in 25 healthy volunteers, using either cellobiose (Ce) (5 g) and mannitol (Ma) (2 g), or lactulose (La) (5 g) and mannitol (2 g), given as oral isosmolar loads. Sugars were recovered in urine collected for 5 h. Analysis was carried out by using anion-exchange chromatography (AEC) with pulsed amperometric detection (PAD). Baseline separation of the above carbohydrates was achieved within 13 min by using a Carbopac PA-100 column and linear gradient elution. Carbohydrate quantification was performed by an internal standard method. The calibration curve for each sugar is linear to 40 mM. The limit of sugar detection is 0.01 mM. Recovery of sugar probes is between 98.2 and 100%.The %La, %Ce, %Ma in urine were evaluated and their ratios (Ce/Ma and La/Ma) were calculated. No significant difference in IP parameters were shown (La/Ma to Ce/Ma 0.018+/-0.014 vs. 0.012+/-0.007; the attendant probability of the null hypothesis being P=0.0714). Ce/Ma and/or La/Ma tests result similarly reliable in the clinical investigation of IP and the described new method is also helpful in urine even with high glucose concentration, without any interference.     

Assessment of the Effect of Intestinal Permeability Probes (Lactulose And Mannitol) and Other Liquids on Digesta Residence Times in Various Segments of the Gut Determined by Wireless Motility Capsule: A Randomised Controlled Trial

Background

Whilst the use of the mannitol/lactulose test for intestinal permeability has been long established it is not known whether the doses of these sugars modify transit time Similarly it is not known whether substances such as aspirin that are known to increase intestinal permeability to lactulose and mannitol and those such as ascorbic acid which are stated to be beneficial to gastrointestinal health also influence intestinal transit time.

Methods

Gastric and intestinal transit times were determined with a SmartPill following consumption of either a lactulose mannitol solution, a solution containing 600 mg aspirin, a solution containing 500 mg of ascorbic acid or an extract of blackcurrant, and compared by doubly repeated measures ANOVA with those following consumption of the same volume of a control in a cross-over study in six healthy female volunteers. The dominant frequencies of cyclic variations in gastric pressure recorded by the Smartpill were determined by fast Fourier transforms.

Results

The gastric transit times of lactulose mannitol solutions, of aspirin solutions and of blackcurrant juice did not differ from those of the control. The gastric transit times of the ascorbic acid solutions were significantly shorter than those of the other solutions. There were no significant differences between the various solutions either in the total small intestinal or colonic transit times. The intraluminal pHs during the initial quartiles of the small intestinal transit times were lower than those in the succeeding quartiles. This pattern did not vary with the solution that was consumed. The power of the frequencies of cyclic variation in intragastric pressure recorded by the Smartpill declined exponentially with increase in frequency and did not peak at the reported physiological frequencies of gastric contractile activity.

Conclusions

Whilst the segmental residence times were broadly similar to those using other methods, the high degree of variation between subjects generally precluded the identification of all but gross variation between treatments. The lack of any differences between treatments in either total small or large intestinal transit times indicates that the solutions administered in the lactulose mannitol test of permeability had no consistent influence on the temporal pattern of absorption. The negatively exponential profile and lack of any peaks in the frequency spectra of cyclic variation in gastric intraluminal pressure that were consistent with reported physiological frequencies of contractile activity profile suggests that the principal source of this variation is stochastic likely resulting from the effects of external events occasioned by normal daily activities on intra-abdominal pressure.

Trial Registration

Australian New Zealand Clinical Trials Registry ACTRN12615000596505