Structural identification and combination mechanism of iron (II)-chelating Atlantic salmon (Salmo salar L.) skin active peptides.

In order to utilize salmon skin for high value, and investigate the structural identification and combination mechanism of iron (II)-chelating peptides systemically, Atlantic salmon (Salmo salar L.) skin, a by-product of Atlantic salmon processing, was treated by two-step enzymatic hydrolysis to obtain salmon skin active peptides (SSAP). Then they reacted with iron (II) to obtain iron (II)-chelating salmon skin active peptides (SSAP-Fe) with a high iron (II) chelating ability of 98.84%. The results of Fourier transform infrared spectroscopy (FTIR), circular dichroism (CD) spectroscopy, 8-anilino-1-naphthalenesulfonic acid ammonium salt hydrate (ANS) combined fluorescence measurement, isothermal titration calorimetry (ITC) and full wavelength ultraviolet (UV) scanning showed that the structural characteristics of SSAP changed before and after chelating iron (II). Reverse phase high performance liquid chromatography (RP-HPLC) and mass spectrometry were used to identify and quantify the peptides in SSAP-Fe. Four peptide sequences (STEGGG, GIIKYGDDFMH, PGQPGIGYDGPAGPPGPPGPPGAP and QNQRESWTTCRSQSSLPDG) were identified. The content of PGQPGIGYDGPAGPPGPPGPPGAP was the highest, at 25.17 µg/mg. The pharmacokinetic and pharmacodynamic properties of these four peptides were also investigated, and the results indicated that they have satisfactory predicted ADMET properties. Molecular docking technology was used to analyze the binding sites between iron (II) and SSAP, and it was found that PGQPGIGYDGPAGPPGPPGPPGAP had the lowest predicted binding energy with iron (II) and the most stable predicted binding energy with iron (II). This results showed that the stability of SSAP-Fe were closely related to the number of covalent bonds and the types of amino acids. This study revealed the structure and combination mechanism of SSAP-Fe, and indicated that SSAP-Fe prepared by chelation may be used as a Fe supplement that can be applied in functional foods or ingredients.

Wheat peptides inhibit the activation of MAPK and NF-κB inflammatory pathways and maintain epithelial barrier integrity in NSAID-induced intestinal epithelial injury.

The use of non-steroidal anti-inflammatory drugs (NSAIDs) has negative effects on the gastrointestinal tract, but the proton pump inhibitors currently in use only protect against gastrointestinal disease and may even make NSAID-induced enteropathy worse. Therefore, new approaches to treating enteropathy are required. This study aimed to investigate the protective effect of wheat peptides (WPs) against NSAID-induced intestinal damage in mice and their mechanism. Here, an in vivo mouse model was built to investigate the protective and reparative effects of different concentrations of WPs on NSAID-induced intestinal injury. WPs ameliorated NSAID-induced weight loss and small intestinal tissue damage in mice. WP treatment inhibited NSAID-induced injury leading to increased levels of oxidative stress and expression levels of inflammatory factors. WPs protected and repaired the integrity and permeability injury of the intestinal tight junction induced by NSAIDs. An in vitro Caco-2 cell model was built with lipopolysaccharide (LPS). WP pretreatment inhibited LPS-induced changes in the Caco-2 cell permeability and elevated the levels of oxidative stress. WPs inhibited LPS-induced phosphorylation of NF-κB p65 and mitogen-activated protein kinase (MAPK) signaling pathways and reduced the expression of inflammatory factors. In addition, WPs increased tight junction protein expression, which contributed to improved intestinal epithelial dysfunction. Our results suggest that WPs can ameliorate NSAID-induced impairment of intestinal barrier functional integrity by improving intestinal oxidative stress levels and reducing inflammatory factor expression through inhibition of NF-κB p65 and MAPK signaling pathway activation. WPs can therefore be used as potential dietary supplements to reduce NSAID-induced injury of the intestine.

Effects of food-derived oligopeptide iron chelates on liver injury and gut microbiota homeostasis in iron-deficiency anemia female rats: a pilot study.

Iron deficiency (ID) is the biggest cause of anemia. This pilot study aimed to investigate the effects of food-derived oligopeptide iron chelates on ameliorating liver injury and restoring gut microbiota homeostasis in iron-deficiency anemia (IDA) female rats. Female Sprague-Dawley rats at 21 days old were selected and randomly divided into a control group (N = 4) and an ID model group (N = 16). The ID model group was fed an iron-deficient diet containing 4 mg kg-1 iron for 28 days to generate the IDA rat model and then randomly subdivided into four groups (N = 4 for each group): ID group, ferrous sulfate group, marine fish oligopeptide iron chelate (MCOP-Fe) group, and whey protein oligopeptide iron chelate (WPP-Fe) group. Iron supplements were given to rats in the three intervention groups once per day via intragastric administration for three weeks. After iron supplementation, the hemoglobin levels in the three intervention groups were significantly improved, with the MCOP-Fe and WPP-Fe groups returning to normal. The ALT and AST levels in the ID group increased significantly, while levels in all intervention groups decreased to normal levels. Liver glutathione in the WPP-Fe group was increased, while the activity of superoxide dismutase also tended to be higher. In addition, 16S rRNA gene sequencing showed that IDA resulted in changes to intestinal microbiota. After intervention, the WPP-Fe group showed increased alpha diversity of intestinal microbes. Therefore, MCOP-Fe and WPP-Fe may improve the iron status of IDA female rats as well as ameliorate liver damage, with WPP-Fe showing a greater potential in improving gut microbiota imbalance.

Effect of phosphorus fertilizer on phytoextraction using Ricinus communis L. in Cu and Cd co-contaminated soil.

Mining activities have led to Cu and Cd contaminated of surrounding agricultural soil. To decrease the Cu and Cd accumulation in crops, the Ricinus communis L. (castor) has been used for phytoremediation. A pot experiment was served to investigate the effect of phosphate fertilizer (Ca(H2PO4)2) on the growth and Cu/Cd uptake of castor in contaminated soil. The results showed that the application of P fertilizer improved the leaf cell morphology, decreased the malonaldehyde (MDA) content of castor leaves, and increased the plant biomass (28.2-34.2%). Besides, phosphate fertilizer still facilitated accumulation Cu and Cd by castor. The addition of phosphate fertilizer increased the contents of Cu in the root of castor, improved the bioconcentration factor (BCF) of Cu, and observably enhanced the accumulation of Cu (up to 201 µg/plant) in castor. Applying phosphorus increased the percentage of residual Cd, diminished the percentage of acid extractable Cd in soil, and the accumulation of Cd in castor was not significantly increased. These results suggest that phosphorus alleviated the stress of heavy metals on castor leaves and enhanced the accumulation of Cu and Cd in castor by promoting the growth of castor.

Applying phosphate fertilizer effectively alleviated the stress of heavy metals on castor and significantly increased the biomass of castor.The reason of applying phosphorus enhanced the castor uptake Cu and Cd was that phosphorus promoted the growth of castor.Applying phosphorus markedly increased the percentage of residual Cd but diminished the percentage of acid extractable Cd in soil.

Effects of Aerobic Exercise Combined with Oyster Peptide Supplement on the Formation of CTX-induced Late-Onset Hypogonadism in Male Rats.

The purpose of this study is to investigate the effect of aerobic exercise (AE) training and/or oyster peptide (OP) supplementation on the formation of late-onset hypogonadism (LOH). AE training and/or OP supplement was performed during Cytoxan (CTX)-induced LOH formation in male SD rats for 6 consecutive weeks. Low dose of CTX could decrease mating times, the levels of luteinizing hormone (LH), total testosterone (TT), free testosterone (FT) in serum and TT, androgen receptor (AR), androgen binding protein (ABP), and glutathione peroxidase (GSH-Px) in testicle, but increase capture latency, mating latency, and malondialdehyde, and downregulate the mRNA expression of steroidogenic acute regulatory (StAR), P450 cholesterol side chain cleavage enzyme (P450scc), and StAR-related lipid transfer domain 7 (StARD7) in testicle. Every change was altered by AE training combined with OP supplement significantly, except for serum LH. Moreover, the effect of AE training combined with OP supplement was better than that of AE training on serum TT, FSH, testicular TT, mating latency, capture times, and mating times. AE training combined with OP supplement during CTX-induced LOH formation can prevent the LOH development by enhancing pituitary-gonads axis's function and reducing testicular oxidative stress to promote testosterone synthesis and spermatogenesis.

[Clinical research of Pinggan Jiangya decoction combined with penetrating needling at Baihui (GV20) in a period of day from 7 am to 9 am in the treatment of grade 1 and 2 essential hypertension].

OBJECTIVE: To explore the clinical effect of Pinggan Jiangya decoction combined with penetrating needling at Baihui (GV20) in a period of day from 7 am to 9 am in the treatment of grade 1 and 2 essential hypertension (EH). METHODS: A total of 150 cases of grade 1 and 2 EH patients were randomized into an observation group and a control group, 75 cases in each group. In the control group, Pinggan Jiangya decoction was prescribed for oral administration one dose a day, while in the observation group, on the basis of the treatment as the control group, penetrating needling was exerted at GV20 once daily. The treatment duration was 8 weeks. Before and after treatment, the TCM syndrome score, 24 h average systolic blood pressure (24 h ASBP), 24 h average diastolic blood pressure (24 h ADBP), 24 h average pulse pressure difference (24 h PP), morning blood pressure surge (MBPS), 24 h SBP variability (24 h SBPV), 24 h DBP variability (24 h DBPV), serum levels of 5-hydroxytryptamine (5-HT) and melatonin (MT) were compared in the patients of the two groups. The clinical therapeutic effect was observed in the two groups. RESULTS: After the treatment, in the self-comparison of each group, the scores of headache, vertigo, backache, soft knees, tinnitus, 24 h ASBP， 24 h ADBP, 24 h PP, MBPS, 24 h SBPV and 24 h DBPV in the two groups were lower than those before treatment (P<0.01), and the above indexes in the observation group were lower than those in the control group (P<0.01). The level of serum 5-HT after the treatment was lower than that of before the treatment (P<0.01), while the level of MT was higher than that of before the treatment (P<0.01) in both two groups, and the level of 5-HT in the observation group was lower than that of the control group, while the level of MT was higher than that of the control group (P<0.01). The total effective rate of the observation group was 93.3% (70/75), better than 76.0% (57/75) of the control group (P<0.01). CONCLUSION: Pinggan Jiangya decoction combined with penetrating needling at GV20 in a period of day from 7 am to 9 am can regulate the levels of serum MT and 5-HT, effectively reduce blood pressure, improve blood pressure variability, control morning peak blood pressure, and has a remarkable effect in the treatment of grade 1 and 2 EH.

Role of Dorsomedial Hypothalamus GABAergic Neurons in Sleep-Wake States in Response to Changes in Ambient Temperature in Mice.

Good sleep quality is essential for maintaining the body's attention during wakefulness, which is easily affected by external factors such as an ambient temperature. However, the mechanism by which an ambient temperature influences sleep-wake behaviors remains unclear. The dorsomedial hypothalamus (DMH) has been reported to be involved in thermoregulation. It also receives projection from the preoptic area, which is an important region for sleep and energy homeostasis and the suprachiasmatic nucleus-a main control area of the clock rhythm. Therefore, we hypothesized that the DMH plays an important role in the regulation of sleep related to ambient temperatures. In this study, we found that cold exposure (24/20/16/12 °C) increased wakefulness and decreased non-rapid eye movement (NREM) sleep, while warm exposure (32/36/40/44 °C) increased NREM sleep and decreased wakefulness compared to 28 °C conditions in wild-type mice. Then, using non-specific and specific apoptosis, we found that lesions of whole DMH neurons and DMH γ-aminobutyric acid (GABA)-ergic neurons induced by caspase-3 virus aggravated the fluctuation of core body temperature after warm exposure and attenuated the change in sleep-wake behaviors during cold and warm exposure. However, chemogenetic activation or inhibition of DMH GABAergic neurons did not affect the sleep-wake cycle. Collectively, our findings reveal an essential role of DMH GABAergic neurons in the regulation of sleep-wake behaviors elicited by a change in ambient temperature.

Clinical Study of Stellate Ganglion Block Combined with General Anesthesia on Hemodynamics, Cognitive Function, and Gastrointestinal Function in Elderly Patients Undergoing Partial Hepatectomy.

Partial hepatectomy under general anesthesia is prone to hemodynamic alterations, and stress reactions are the main contributing factors to postoperative cognitive function in elderly partial hepatectomy patients. Postoperative cognitive dysfunction increases the incidence of postoperative complications and long-term morbidity and mortality in elderly patients. With the increasing trend of aging population and the gradual increase of elderly people undergoing surgical treatment, it is especially important to study the corresponding prevention and treatment measures. In this study, a total of 90 patients with primary liver cancer who received hepatectomy in our hospital from July 2020 to July 2021 were included as the research subject. The changes in hemorheology, stress-related indexes, cognitive function, postoperative pain, and gastrointestinal function were compared between the two groups The results showed that SGB combined with general anesthesia can effectively reduce hemodynamic fluctuations in elderly partial hepatectomy patients, alleviate surgical stress, promote postoperative recovery of cognitive function and gastrointestinal function with high safety, and is worthy of clinical promotion.

Optical Stimulation of Thalamic Spindle Circuitry Sustains Electroencephalogram Patterns of General Anesthesia but not Duration of Loss of Consciousness.

Alterations in thalamic GABAergic signaling are implicated in mediating the rise in 12-30 Hz electroencephalogram (EEG) activity that signals anesthetic-induced loss-of-consciousness with GABAA receptor-targeting general anesthetics. A number of modeling studies have identified that anesthetic-induced alterations in thalamocortico-corticothalamic signaling in the same network that generates sleep spindles would be sufficient to elicit this key EEG signature of anesthetic hypnosis with general anesthetic agents. Accordingly, we hypothesize that targeted stimulation of this thalamic GABAergic circuitry into a sleep-spindle mode of activity would promote the general anesthetic effects of etomidate. We recorded EEG activity and loss-of-righting reflex in transgenic mice expressing channel rhodopsin-2 on GABAergic neurons (ChR2-VGAT, n = 8) and control, wild-type mice (C57BL/6J, n = 8). On two consecutive days mice were randomly assigned to receive spindle-rhythm stimulation via an optical probe targeting the left reticular thalamic nucleus or no stimulation. After an initial 30-minute recording, mice were administered etomidate (12 mg/kg, intraperitoneal) and recorded for 90 min with or without optical stimulation. Etomidate elicited an increase in 12-30 Hz EEG power in wild-type and ChR2-VGAT mice for 20 min following administration (p < 0.001). Optical spindle-rhythm stimulation prolonged the increase in 12-30 Hz activity in ChR2-VGAT mice only (p = 0.023). Spindle-rhythm stimulation also increased the incidence and duration of sleep spindle-like oscillations in ChR2-VGAT mice only (all p ≤ 0.001). Despite the maintained anesthetic-like changes in EEG activity, optical spindle-rhythm stimulation was not associated with changes in the time to and duration of the loss-of-righting reflex, a behavioral endpoint of etomidate-induced general anesthesia in rodents.

Postoperative intussusceptions in children and infants: a systematic review.

BACKGROUND: Postoperative intussusception (POI) is an unusual complication in children and infants who underwent various kinds of surgery. The early recognition was difficult for its rarity and atypical presentations. This study evaluates the clinical features of POI through a literature review. METHODS: MEDLINE database was searched for relevant articles that reported the children and infants with POI since 1990 in English-language using the key word "postoperative intussusception". All published studies containing clinical data for POI in children and infants were included. Reference lists of retrieved articles were reviewed for additional cases. Detailed data of the included cases were extracted and analyzed. RESULTS: Twenty-six studies with total 127 cases of POI were included. According to the extracted data, the median age was 19 months with the male-to-female ratio 1.5:1. There were 65 operations (51.2 %) that involved gastrointestinal system, 26 cases (20.5 %) of retroperitoneal tumor resection, 12 operations (9.4 %) involved diaphragm, 8 operations (6.3 %) involved urinary system, 5 cases (3.9 %) of partial pancreatectomy, 11 cases (8.7 %) of non-abdominal operations. 75.5 % presented symptoms in the first 7 days after surgery. The prominent symptom was bilious vomiting or increased nasogastric output (87.1 % of 101 patients), following abdominal distention (74.3 %), abdominal pain (35.6 %). Six cases (5.0 %) of ileocolic POI were reduced successfully by air enema. The small bowel intussusception attributed 85.6 % of POI (95 patients). Laparotomy and manual reduction were performed in 104 cases (86.0 %). Nine patients (7.4 %) underwent intestinal resection and anastomosis. CONCLUSIONS: POI should be suspected in pediatric surgical patients who showed signs of intestinal obstruction in the early postoperative period. Early recognition and prompt management are important.

A simple method for the isolation and purification of resveratrol from Polygonum cuspidatum.

Resveratrol, a polyphenol compound with strong biological activity, has been widely used in medicine, health products and cosmetic industries. It is also the main active component of Polygonum cuspidatum, a well-known traditional Chinese medicine. We developed a simple and effective method for the preparation of resveratrol from P. cuspidatum. The whole preparative process consisted of reflux extraction, filtering, hydrolyzing, liquid-liquid extraction and eluting. Filtering is to remove non polar or less polar compounds and debris fragments from the extract. Hydrolyzing is to transform polydatin to resveratrol to improve the yield of resveratrol. Eluting is to remove impurities including strong acidic and water-soluble compounds. By acid hydrolysis of glycoside (polydatin), the yield of resveratrol increased about 4-fold. The extraction recovery in different stages was high, and the content of resveratrol in the final product was over 73.8%. Compared with other methods reported, this technology is eco-friendly, easier to perform, and also has a lower cost.

[Protective effects of tetrandrine against congenital diaphragmatic hernia in rats].

OBJECTIVE: To study the protective effect of the prenatal use of tetrandrine (TET) against congenital diaphragmatic hernia (CDH) in rats and possible mechanisms. METHODS: Pregnant female Sprague-Dawley rats were randomly divided into 3 groups: control, nitrofen and TET treatment. The later two groups were administered with nitrofen by gavage on day 9.5 of gestation. On day 18.5 of gestation, TET (30 mg/kg) was given by gavage (once a day, for three days) in the TET treatment group. On day 21 of gestation, parts of pregnant rats were delivered by cesarean section and amniotic fluid was collected. The fetal rats were examined for a diaphragmatic hernia. Lung histologic evaluations with microscope and immunohistochemistry staining of TNF-α were performed. TNF-α in amniotic fluid was detected using ELISA. The remaining pregnant rats were allowed to deliver spontaneously at term. The survival of pup rats was observed until 24 hrs of age. RESULTS: In the nitrofen group, significant lung hypoplasia was presented not only in fetuses with CDH but also in those without CDH. Stronger expression of TNF-α was observed in fetal lungs and amniotic fluid in the nitrofen group, even when CDH was absent. The TET treatment group showed improved lung development compared with the nitrofen group. The incidence of large diaphragmatic hernia in the TET treatment group was lower than that in the nitrofen group (P<0.05), and the expression of TNF-α in fetal lungs and amniotic fluid in the TET treatment group was also lower than in the nitrofen group (P<0.01). The 24-hr survival rate of pup rats in the TET group was higher than that in the nitrofen group (P<0.01). CONCLUSIONS: Prenatal use of TET can improve nitrofen-induced pulmonary hypoplasia, decrease the incidence of large diaphragmatic hernia and increase the survival rate of pup rats, possibly through a reduction in the production of TNF-α in fetal lungs and amniotic fluid in rats with CDH.

Effect of prenatal tetrandrine administration on transforming growth factor-beta1 level in the lung of nitrofen-induced congenital diaphragmatic hernia rat model.

PURPOSE: Tetrandrine (Tet) is a bisbenzylisoquinoline alkaloid isolated from the root of Stephania tetrandra, which has been used in traditional Chinese medicine to treat patients with silicosis, asthma, and pulmonary hypertension, and others and can be used as a pulmonary therapeutic agent. We hypothesized that it can also improve the lung growth in congenital diaphragmatic hernia (CDH) for its multiple biological effects. There are increasing evidences that suggest transforming growth factor beta1(TGF-beta1) plays a crucial role in fetal lung growth and morphogenesis. The aim of this study was to evaluate the effect of prenatal administration of Tet and to investigate its possible mechanism on the expression of TGF-beta1 in the lung of nitrofen-induced CDH rat model. METHODS: A CDH model was induced in pregnant Sprague-Dawley rats by administration of nitrofen on day 9.5 of gestation (Ed9.5 term, day 22). Tetrandrine (30 mg/kg) was given through gavage (once a day, for 3 days) on Ed11.5. Accordingly, there were 3 groups as follows: control (n = 9), CDH (n = 9), and CDH + Tet (n = 9). All the fetuses were delivered by cesarean delivery on Ed16.5, 18.5, and 21.5, respectively, to check if diaphragmatic hernia existed on each fetus, then the lung tissue weight (LW) and body weight (BW) of each fetus were recorded. Histologic evaluations and TGF-beta1 immunohistochemistry staining in the lung sample were performed for image analysis. RESULTS: Diaphragmatic hernia was observed in 95 of the 112 rat fetuses in CDH and CDH + Tet groups on Ed18.5 and Ed21.5 (84.8%), the incidence between the 2 groups had no statistical significance (P = .642). Lung weight/body weight in the CDH group and the CDH + Tet group were lower than that in the control group (P < .01), and LW/BW in the CDH group was lower than that in the CDH + Tet group (P < .05). Observed under the light microscope and electron microscope, marked hypoplasia of the lungs in fetuses among the CDH groups was observed, in contrast to improvement of the lungs in CDH + Tet fetuses. Statistical differences in morphological parameters (percentage of alveoli area, counting bronchus) were found even on Ed16.5 when diaphragm had not closed (P < .01). The number of type II pneumocytes and lamellar bodies in each group had no significant difference (P > .05). The immunoreactivity of TGF-beta1 in CDH group and CDH + Tet group were markedly stronger than that in the control group (P < .01). In addition, TGF-beta1 expression in the CDH group was stronger than that in the CDH + Tet group (P < .01). CONCLUSION: Nitrofen can interfere with lung development early in the fetal rat development before and separate from diaphragm development, and increased expression of TGF-beta1 in the lung of CDH rat model may suppress lung growth and development. Prenatal treatment with Tet can improve the growth of the lung of the nitrofen-induced CDH fetuses and its mechanism seems to be involved in downregulating the expression of TGF-beta1. It is a likely new approach to treat CDH and its coexistent lung hypoplasia by maternal Tet administration.

Prenatal tetrandrine treatment can reverse the abnormal conditions in the lung of newborn with congenital diaphragmatic hernia.

Pulmonary hypoplasia and persistent pulmonary hypertension are the most important reasons for the high morbidity and mortality of congenital diaphragmatic hernia (CDH). Despite surgical advances and advances in neonatal intensive care, the mortality still remains high. Then the research on how to improve prenatal fetal lung growth has become a focus. Some researches involved in fetal surgery, tracheal occlusion, prenatal use of corticosteroids etc., have been carried out in CDH animal models and humans. But the results either showed no benefit for the outcome of CDH or were unproved. Tetrandrine is a bisbenzylisoquinoline alkaloid isolated from the root of Stephania tetrandra. It has been used in traditional Chinese medicine for several decades to treat patients with silicosis, asthma and pulmonary hypertension etc. Some researches showed that prenatal tetrandrine administration can improve the lung development in CDH rat models. We hypothesize that prenatal treatment with tetrandrine can reverse the abnormal condition in the lung of newborn with CDH, and thus decrease the mortality.

Tissue distribution and excretion of resveratrol in rat after oral administration of Polygonum cuspidatum extract (PCE).

PURPOSE: Polygonum cuspidatum extract as a traditional Chinese medicine is extracted from the dried rhizome and root of Polygonum cuspidatum Sieb.et Zucc. Resveratrol is one of its active components. Studies were performed in rats to define the tissue distribution and excretion of resveratrol in urine and bile, and to characterize (if possible) any metabolites of resveratrol observed in tissues after ig 20mg/kg Polygonum cuspidatum extract. METHOD: For tissue distribution studies, tissues (300 mg) were homogenized and centrifuged with methanol, and metabolites found in selected tissue extract were identified by LC/MS/MS. For urinary and biliary excretion experiments, urine and bile samples were cleaned up by using solid-phase extraction (SPE) with polyamide cartridges. All the concentrations of resveratrol in these biological samples were determined by HPLC with UV detection. RESULT: After a single oral dose of 20mg/kg PCE in rats, resveratrol was mainly distributed in stomach, duodenum, liver and kidney with detectable metabolites resveratrol monoglucuronide and resveratrol monosulfate. The majority of the resveratrol was excreted as metabolites, only 0.59% and 0.027% of the dosage were excreted in urine and bile respectively as unchanged drug within 24h.

[Study on LC-MS fingerprint for quality assessment of Aristolochia manshuriensis with chemical pattern recognition].

OBJECTIVE: To analyze LC-MS fingerprints of Aristolochia manshuriensis for quality assessment with two different chemical pattern recognition models. METHOD: LC-MS fingerprints of A. manshuriensis were established from 24 batches of samples from different habitats. SIMCA and Clustering analysis were used to compare the parameters of the 29 common peaks. RESULT: Two methods had good consistency, while they reflected the inherent sample information from different perspectives, respectively. CONCLUSION: Modern equipment analysis technology and multivariable chemical pattern recognition would be an efficient way for quality control and variety identification of A. manshuriensis.

[Effect of tetrandrine on lung development: experiment of nitrofen-induced congenital diaphragmatic hernia fetal rat model].

OBJECTIVE: To assess the efficacy of prenatal administration of tetrandrine (TET) on pulmonary hypoplasia in the nitrofen-induced congenital diaphragmatic hernia (CDH) fetal rat model. METHODS: Six timed-pregnant female Sprague-Dawley rats were randomly divided into 3 equal groups: CDH group (receiving gavage of nitrofen 125 mg dissolved in seed fat on day 9.5), and TET group (receiving gavage of nitrofen 125 mg on day 9.5 and then gavage of TET 30 mg/kg on days 11.5 - 14.5), and control group (given the same dose of peanut oil on day 9.5 and the same dose of normal saline on days 11.5 - 14.5). The fetuses were delivered by cesarean section on day 21 to undergo light microscopy and electron microscopy. The numbers of type II pneumocytes were recorded and compared. RESULTS: CDH were detected in 32 of the 41 fetuses from the CDH and TET groups with a teratogenic rate of 78%, however, without a significant difference between the CDH and TET groups (P = 0.645). Microscopy showed significant lung hypoplasia in both histologic structure and cellular structure in the CDH group; however the lung development of the TET group was improved in comparison to the CDH group. There was no significant difference in numbers of type II pneumocytes among the 3 groups (P = 0.779). CONCLUSION: Prenatal administration of TET can improve the lung development of CDH rats in both histological structure and cellular structure. This may provide a new idea for the clinical treatment of CDH.

Effect of antenatal tetrandrine administration on endothelin-1 and epidermal growth factor levels in the lungs of rats with experimental diaphragmatic hernia.

PURPOSE: The aim of this study was to evaluate the effect of the traditional Chinese medicine tetrandrine (Tet) and to determine its possible mechanism on expression of endothelin-1 (ET-1) and epidermal growth factor (EGF) in the lung of a rat model of nitrofen-induced congenital diaphragmatic hernia (CDH). METHODS: A single oral dose (115 mg/kg) of nitrofen on day 9.5 of pregnancy was maternally administered to induce CDH. Pregnant rats were divided into 4 groups on day 18.5: control (n = 5), CDH (n = 5), CDH+dexamethasone (Dex) (n = 5), and CDH+Tet (n = 5). All fetuses were delivered by cesarean delivery on day 21.5. Accordingly, there were 4 groups of fetuses: control (n = 38), CDH (n = 25), CDH+Dex (n = 21), and CDH+Tet (n = 22). Lung tissue weight (LW) and body weight (BW) of each fetus were recorded, lung histologic evaluations and ET-1 and EGF immunohistochemistry staining were performed, and image analysis was performed after lung processing. RESULTS: Five female rats in the control group produced 38 fetuses without CDH. CDH was observed in 68 of the 128 rat fetuses (53.1%) among the other 3 groups. The LW/BW ratio of the CDH group was significantly lower than those of the Dex and EGF groups (P < .05). The lungs of fetuses with CDH showed marked abnormal structure such as pulmonary hypoplasia and vascular remodeling, in contrast to improved pulmonary structure in lungs of fetuses in the CDH+Dex and CDH+Tet groups. Statistical differences in morphologic parameters (radial alveolar counts, percentage of alveoli, percentage of medial wall thickness, and vascular volume) were found (P < .05). The immunoreactivity of EGF and ET-1 in the CDH group was markedly stronger than that in the control, CDH+Dex, and CDH+Tet groups (P < .01). In addition, EGF and ET-1 expression in the CDH+Dex and CDH+Tet groups was stronger than that in the control group (P < .05). There was no difference in lung EGF and ET-1 immunoreactivity between CDH+Dex and CDH+Tet groups (P > .05). CONCLUSION: Antenatal treatment with Tet may improve lung growth and vascular remodeling, and its mechanism seems to be involved in decreasing EGF and ET-1 expression. Tet administered maternally may be a hopeful new therapeutic option in the treatment of CDH and may be effective in helping to avoid the side effects of Dex.

[Effect and significance of tetrandrine on epidermal growth factor and its receptor in the lung of congenital diaphragmatic hernia rat model].

OBJECTIVE: To investigate the effect of the traditional Chinese medicine "Tetrandrine"(TET) and its significance on epidermal growth factor (EGF) and its receptor (EGFR) in the lung of nitrofen-induced congenital diaphragmatic hernia (CDH) rat model. METHODS: Twenty female rats were given maternal administration of a single oral dose (115 mg/rat) of nitrofen to induce CDH at 9.5 days after pregnancy and were divided into normal solution group (NS, n=5), dexamethasone group (Dex, n=5), tetrandrine group (TET, n=5) and Dex+TET group (n=5) at 18.5 days; 4 rats were given edible oil as controls. All fetuses were delivered by cesarean section at 21.5 days. Lung histologic evaluations and EGF, EGFR immunohistochemical staining and image analysis were performed. RESULTS: CDH was observed in 64 of the 137 rat fetuses (46.7%) in the experimental groups; no CHD was observed in 36 rat fetuses of control group. The lungs of CDH fetuses showed marked hypoplasia in NS group, in contrast to improved mesenchymal differentiation in that of Dex, TET, Dex+TET groups. The expression of EGF was weaker and weaker and that of EGFR was stronger and stronger as following order: NS, TET, DEX, T+D and control groups; showing significant differences between them (P<0.05). CONCLUSION: Prenatal TET administration shows marked improvement in pulmonary hypoplasia through pre-regulating crest-time of EGF expression and up-regulating EGFR expression in the lungs of nitrofen-induced CDH rat model. A combination of TET and Dex would generate evident synergistic effect.

High performance liquid chromatography - tandem mass spectrometric determination of cyclovirobuxine D in human plasma.

A sensitive, specific and rapid high performance liquid chromatography - tandem mass spectrometric (LC/MS/MS) method for the determination of cyclovirobuxine D in human plasma was developed and validated. The triple-quadrupole tandem mass spectrometric detector with an electrospray interface (ESI) was operated under the selected reaction monitoring (SRM) mode. After the addition of citalopram as an internal standard (IS), plasma samples were extracted with ethyl acetate. Chromatographic separation of the analytes was performed on a Kromasil CN column with a mobile phase of methanol/water (88/12, v/v) containing 0.4% formic acid. Linearity was established for the range of concentration 0.2-40ng/ml. Under optimized conditions, the mean recovery was 86.6%. The intra-day precision ranged from 4.56% to 7.81%, while the intra-day accuracy ranged from 2.75% to 11.0%. The inter-day precision was in the range 3.87-10.7%, and the inter-day accuracy was in the range -4.00% to 2.50%. The cyclovirobuxine D was stable in human plasma after three freeze-thaw cycles, under storage at room temperature for 12h, in a freezer at -20 degrees C for 15 days and during processing (in autosampler) at 10 degrees C for 24h. The validated method is suitable for quantitative determination of cyclovirobuxine D in human plasma in pharmacokinetics study and has been successfully applied to the analysis of clinical samples.