Zinc alleviates thermal stress-induced damage to the integrity and barrier function of cultured chicken embryonic primary jejunal epithelial cells via the MAPK and PI3K/AKT/mTOR signaling pathways.

Zinc (Zn) could alleviate the adverse effect of high temperature (HT) on intestinal integrity and barrier function of broilers, but the underlying mechanisms remain unclear. We aimed to investigate the possible protective mechanisms of Zn on primary cultured broiler jejunal epithelial cells exposed to thermal stress (TS). In Exp.1, jejunal epithelial cells were exposed to 40℃ (normal temperature, NT) and 44℃ (HT) for 1, 2, 4, 6, or 8 h. Cells incubated for 8 h had the lowest transepithelial resistance (TEER) and the highest phenol red permeability under HT. In Exp.2, the cells were preincubated with different Zn sources (Zn sulfate as iZn and Zn proteinate with the moderate chelation strength as oZn) and Zn supplemental levels (50 and 100 µmol/L) under NT for 24 h, and then continuously incubated under HT for another 8 h. TS increased phenol red permeability, lactate dehydrogenase (LDH) activity and p-PKC/PKC level, and decreased TEER, cell proliferation, mRNA levels of claudin-1, occludin, zona occludens-1 (ZO-1), PI3K, AKT and mTOR, protein levels of claudin-1, ZO-1 and junctional adhesion molecule-A (JAM-A), and the levels of p-ERK/ERK, p-PI3K/PI3K and p-AKT/AKT. Under HT, oZn was more effective than iZn in increasing TEER, occludin, ZO-1, PI3K, and AKT mRNA levels, ZO-1 protein level, and p-AKT/AKT level; supplementation with 50 µmol Zn/L was more effective than 100 µmol Zn/L in increasing cell proliferation, JAM-A, PI3K, AKT, and PKC mRNA levels, JAM-A protein level, and the levels of p-ERK/ERK and p-PI3K/PI3K; furthermore, supplementation with 50 µmol Zn/L as oZn had the lowest LDH activity, and the highest ERK, JNK-1, and mTOR mRNA levels. Therefore, supplemental Zn, especially 50 µmol Zn/L as oZn, could alleviate the TS-induced integrity and barrier function damage of broiler jejunal epithelial cells possibly by promoting cell proliferation and tight junction protein expression via the MAPK and PI3K/AKT/mTOR signaling pathways.

Lycopene improves maternal reproductive performance by modulating milk composition and placental antioxidative and immune status.

Placental health and milk quality are important for maternal reproductive performance during pregnancy and lactation. Lycopene plays an important role in antioxidation, anti-inflammation and regulating lipid metabolism. The goal of the present study was to investigate the effects of dietary lycopene supplementation in the pig model on reproductive performance, placental health and milk composition during maternal gestation and lactation. In the present study, the litter size of live piglets was increased and the litter size of dead piglets was decreased by lycopene supplementation of the diet of sows. The litter weight at birth and weaning were increased in the lycopene group. Through placental proteomics, we enriched differentially expressed proteins (DEPs), gene ontology (GO) terms, and Kyoto encyclopedia of proteins and genomes (KEGG) pathways involved in immunity, anti-inflammation, antioxidants, and lipid metabolism and transport. Furthermore, in terms of placental health, we analyzed the levels of related enzymes, metabolites and mRNA expression in the placenta. Lycopene was shown to reduce mRNA expression and the levels of placental inflammatory factors, increase the content of immunoglobulin, improve the antioxidant capacity, and improve lipid metabolism and lipid transport in the placenta. In terms of sow milk composition, lycopene increased the levels of immunoglobulins in colostrum and lactose in colostrum and milk. Overall, the results of the present study demonstrate that dietary lycopene supplementation of sows during gestation and lactation improves the reproductive performance to a certain extent; this may be due to lycopene improving the placental health and milk composition of sows.

Clinical Efficacy of Carbon Dioxide Laser Combined with ALA Photodynamics in the Treatment of Condyloma Acuminatum.

'Purpose. To observe the clinical efficacy and safety of carbon dioxide laser combined with ALA photodynamics in the treatment of condyloma acuminatum. Method. A total of 211 patients with condyloma acuminatum admitted to our hospital from April 2018 to June 2021 were selected as the observation object. They were divided into the intervention group (CO2 laser combined with ALA photodynamic therapy, 125 cases) and conventional group (CO2 laser treatment, 86 cases), and the efficacy and incidence of adverse reactions between the two groups were compared. Result. The total effective rate of the intervention group (96.00%) was significantly higher than that of the conventional group (84.88%) (P < 0.05). The total incidence of adverse reactions in the intervention group (8.00%) was lower than that in the conventional group (32.56%) (P < 0.05). Univariate analysis showed that the patient's smoking history, drinking history, course of disease, wart area, and number of sexual partners were related to the short-term prognosis (P < 0.05). Multivariate logistic regression analysis showed that the patient's course of disease, the area of the wart body, and the number of sexual partners were independent factors affecting the prognosis of patients with condyloma acuminatum (P < 0.05). Conclusion. Carbon dioxide laser combined with ALA dynamics treatment of condyloma acuminatum significantly improves the clinical efficacy, does not increase the incidence of adverse reactions, and has important clinical therapeutic value. The course of the disease, the area of the wart, and the number of sexual partners are independent factors affecting the prognosis of patients with condyloma acuminatum.

Lycopene modulates lipid metabolism in rats and their offspring under a high-fat diet.

The purpose of this study was to investigate the effects of lycopene supplementation on lipid metabolism in rats and their offspring. The experiment was conducted on 60 female rats divided into four groups: normal diet, normal diet with 200 mg kg-1 lycopene, high-fat diet, and high-fat diet with 200 mg kg-1 lycopene. The plasma levels of TG, LDL-C, AST and ALT in female rats fed a high-fat diet were significantly increased (P < 0.05). Lycopene supplementation reduced the plasma TG, LEP and AST levels (P < 0.05). In addition, the activity of ACC and mRNA expression of SREBP1c, FAS, PPARγ, CPT1, HMGCR, ACC, PLIN1 and FATP1 in the liver were also increased after feeding a high-fat diet (P < 0.05), whereas the expression of HSL was decreased (P < 0.05). Lycopene increased the activity of HSL and the expression of ATGL in the liver (P < 0.05), and the activity of ACC and mRNA expression of HMGCR and ACC were decreased (P < 0.05). For the offspring, maternal feeding of a high-fat diet reduced the plasma HDL-C levels (P < 0.05), but lycopene supplementation reduced the plasma TC levels (P < 0.05). Maternal high-fat diet also decreased the activity of HSL and the expression of CD36, PLIN1 and FATP1 in the liver while increasing the expression of PPARγ (P < 0.05). Maternal lycopene supplementation decreased the activities of ACC and FAS in the liver and decreased the expression of PPARγ, ACC and PLIN1 (P < 0.05). Maternal feeding of a high-fat diet increased the level of oxidative stress in the liver, the level of blood lipids in plasma and the rate of lipid production in the liver of rats and their offspring. Maternal lycopene supplementation can reduce the level of oxidative stress in rats and their offspring, reduce the level of blood lipids in plasma, and also reduce the rate of lipid production in the liver of rats and offspring.

Quantitative proteomics analysis of Fructus Psoraleae-induced hepatotoxicity in rats.

Fructus Psoraleae, which is commonly consumed for the treatment of osteoporosis, bone fracture, and leucoderma, induces liver injury. This study investigated the pathogenesis of the ethanol extract of Fructus Psoraleae (EEFP)-induced liver injury in rats. EEFP (1.35, 1.80, and 2.25 g·kg-1) was administrated to Sprague Dawley (SD) rats for 30 d. We measured liver chemistries, histopathology, and quantitative isobaric tags for relative and absolute quantitation (iTRAQ)-based protein profiling. EEFP demonstrated parameters suggestive of liver injury with changes in bile secretion, bile flow rate, and liver histopathology. iTRAQ analysis showed that a total of 4042 proteins were expressed in liver tissues of EEFP-treated and untreated rats. Among these proteins, 81 were upregulated and 32 were downregulated in the treatment group. KEGG pathway analysis showed that the drug metabolic pathways of cytochrome P450, glutathione metabolism, glycerolipid metabolism, and bile secretion were enriched with differentially expressed proteins. The expression of key proteins related to the farnesoid X receptor (FXR), i.e., the peroxisome proliferators-activated receptor alpha (PPAR-α), were downregulated, and multidrug resistance-associated protein 3 (MRP3) was upregulated in the EEFP-treated rats. Our results provide evidence that EEFP may induce hepatotoxicity through various pathways. Furthermore, our study demonstrates changes in protein regulation using iTRAQ quantitative proteomics analysis.

Pyrroloquinoline quinone inhibits the production of inflammatory cytokines via the SIRT1/NF-κB signal pathway in weaned piglet jejunum.

The small intestine is an important digestive organ and plays a vital role in the life of a pig. In this study, we explored the regulatory role and molecular mechanism of pyrroloquinoline quinone (PQQ) on intestinal health and to discussed the interaction between PQQ and vitamin C (VC). A total of 160 healthy piglets weaned at 21 d were randomly divided into four treatment groups according to 2 × 2 factoring. The results showed that dietary PQQ could significantly decrease the levels of plasma globulin, albumin/globulin (A/G), indirect bilirubin (IBIL), blood urea nitrogen (BUN), creatinine (CREA) (P < 0.05 for each), total bilirubin, (TBIL) (P < 0.01), diamine oxidase (DAO) (P < 0.01) and immunoglobulin G (IgG) (P < 0.0001) and increase the levels of immunoglobulin A (IgA) and immunoglobulin M (IgM) (P < 0.0001) in the plasma of weaned piglets. Similarly, dietary VC could significantly decrease the levels of plasma globulin, A/G, DAO (P < 0.05 for each) and IgG (P < 0.0001) and increase the levels of IgA and IgM (P < 0.0001) in the plasma of weaned piglets. In addition, dietary PQQ increased (P < 0.05) the mRNA levels of antioxidant genes (NQO1, UGT1A1, and EPHX1), thereby enhancing (oxidized) nicotinamide adenine dinucleotide (NAD+) concentration and sirtuin 1 (SIRT1) activity in tissues. However, the addition of 200 mg kg-1 VC to the diet containing PQQ reduced most of the effects of PQQ. We further show that PQQ reduced (P < 0.05) the expression of inflammation-related genes (IL-2, IL-6, TNF-α, and COX-2) via the SIRT1/NF-κB deacetylation signaling. In conclusion, our data reveals that PQQ exerts a certain protective effect on the intestines of piglets, but higher concentrations of VC react with PQQ, which inhibits the regulatory mechanism of PQQ.

In vitro study of the nephrotoxicity of total Dahuang (Radix Et Rhizoma Rhei Palmati) anthraquinones and emodin in monolayer human proximal tubular epithelial cells cultured in a transwell chamber.

OBJECTIVE: To examine changes in the morphology and physiological functions of human proximal tubular epithelial cells (HK-2 cells) caused by total Dahuang (Radix Et Rhizoma Rhei Palmati) anthraquinones (TDA) and emodin. METHODS: HK-2 cells were cultured on polycarbonate (PCF) membranes to form a complete monolayer of cells. A fluorescein isothiocyanate-dextran (FITC) permeability assay was conducted and secretion of γ-glutamyltranspeptidase (GGT), lactate dehydrogenase (LDH), N-acetyl-ß-D-glucosaminidase (NAG) and kidney injury molecule 1 (KIM-1) was examined. The reabsorption of glucose and the excretion of para-aminohippuric acid (PAH) by HK-2 cells were also examined. The morphology of HK-2 cells was observed using optical microscopy and scanning electron microscopy. The cytoskeleton of HK-2 cells was observed under a fluorescence microscope. RESULTS: Compared with the results for the dimethyl sulfoxide group, treatment of cells with TDA and emodin showed statistically significant differences in the FITC leakage rate, the apical / basolateral ratio of LDH and GGT, and the secretion of GGT, LDH, NAG and KIM-1. At 64 µg/mL, TDA markedly inhibited blood glucose reabsorption and remarkably suppressed PAH excretion by HK-2 cells. Both TDA and emodin caused various degrees of damage to the morphology and cytoskeleton of HK-2 cells with the degree of damage correlating positively with the dosage of the tested substances. CONCLUSION: Both TDA and emodin caused damage to human renal proximal tubular epithelial cells at certain dosages. At the same dosage, TDA caused more severe damage than emodin to the HK-2 cells.

Reproductive role of miRNA in the hypothalamic-pituitary axis.

The hypothalamic-pituitary-gonadal (HPG) axis plays a critical role in regulating reproductive function. Gonadotropin-releasing hormone (GnRH), which is secreted by the hypothalamus, acts on pituitary gonadotrophs to stimulate luteinizing hormone (LH) and follicle-stimulating hormone (FSH) synthesis and secretion, ultimately affecting the animal's fertility. MicroRNAs are small, non-coding RNAs that are widely expressed throughout the brain and can fine-tune gene expression post-transcriptionally. Recently, growing evidence has unveiled the central position of miRNAs within a key regulatory process involving GnRH secretion and subsequent activation in the pituitary. Although transcriptional regulation of reproduction has been well studied, the post-transcriptional processes are less well understood. In this review, we elaborate comprehensively on the critical role of miRNAs in the reproductive process, including both temporal and spatial aspects. A better understanding of how miRNAs impact the neuroendocrine system may improve our knowledge of reproduction and provide novel targets for therapeutic development.

Fructus phyllanthi tannin fraction induces apoptosis and inhibits migration and invasion of human lung squamous carcinoma cells in vitro via MAPK/MMP pathways.

AIM: Fructus phyllanthi tannin fraction (PTF) from the traditional Tibetan medicine Fructus phyllanthi has been found to inhibit lung and liver carcinoma in mice. In this study we investigated the anticancer mechanisms of PTF in human lung squamous carcinoma cells in vitro. METHODS: Human lung squamous carcinoma cell line (NCI-H1703), human large-cell lung cancer cell line (NCI-H460), human lung adenocarcinoma cell line (A549) and human fibrosarcoma cell line (HT1080) were tested. Cell viability was detected with MTT assay. Cell migration and invasion were assessed using a wound healing assay and a transwell chemotaxis chambers assay, respectively. Cell apoptosis was analyzed with flow cytometric analysis. The levels of apoptosis-related and metastasis-related proteins were detected by Western blot and immunofluorescence. RESULTS: PTF dose-dependently inhibited the viability of the 3 human lung cancer cells. The IC50 values of PTF in inhibition of NCI-H1703, NCI-H460, and A549 cells were 33, 203, and 94 mg/L, respectively. PTF (15, 30, and 60 mg/L) dose-dependently induced apoptosis of NCI-H1703 cells. Treatment of NCI-H1703 and HT1080 cells with PTF significantly inhibited cell migration, and reduced the number of invasive cells through Matrigel. Furthermore, PTF dose-dependently down-regulated the expression of phosphor-ERK1/2, MMP-2 and MMP-9, up-regulated the expression of phosphor-JNK, but had no significant effect on the expression of ERK1/2 or JNK. CONCLUSION: PTF induces cell apoptosis and inhibits the migration and invasion of NCI-H1703 cells by decreasing MPPs expression through regulation of the MAPK pathway.

Effects of hypothermia on S100B and glial fibrillary acidic protein in asphyxia rats after cardiopulmonary resuscitation.

The aim of the study was to investigate the effects of hypothermia on S100B and glial fibrillary acidic protein (GFAP) in serum and hippocampus CA1 area in asphyxiated rats after cardiopulmonary resuscitation (CPR). A total of 100 SD rats were designated into four groups: group A, sham operation group; group B, rats received conventional resuscitation; group C, rats received conventional resuscitation and hypothermia at cardiac arrest; group D, rats received conventional resuscitation and hypothermia at 30 min after restoration of spontaneous circulation (ROSC). Rats were then killed by cardiac arrest at 2 and 4 h after ROSC; brain tissue was taken to observe dynamic changes of S100B and GFAP in serum and hippocampus CA1 area. Following ROSC, S100B levels increased from 2 to 4 h in group B, C, and D. In addition, S100B in serum and hippocampus CA1 area was all significantly increased at different time points compared with group A (P < 0.05). Following ROSC, serum S100B level at 2 h in group C was significantly decreased compared with group B, but the difference was not statistically significant (P > 0.05). Moreover, S100B in serum at 4 h after ROSC was significantly decreased (P < 0.05), S100B in cortex was significantly decreased (P < 0.05). The expression of GFAP was also examined. GFAP level in hippocampus CA1 area was significantly decreased in group B, C, and D at 4 h after ROSC compared with group A (P < 0.05). S100B and GFAP were expressed in rat serum and hippocampus CA2 area at early stage after ROSC, which can be used as sensitive markers for brain injury diagnosis and prognosis prediction. Hypothermia is also shown to reduce brain injury after CPR.

[Effect of total flavones of Epimedium leptorrhizum on osteoporosis in ovariectomized rats].

OBJECTIVE: To observe the effect of total flavones of Epimedium leptorrhizum (YYH-C) on osteoporosis in ovariectomized rats. METHOD: Ovariectomized female rats were randomly divided into the model group, YYH-C lower, middle and high dose (0.7, 1.4, 2.8 g x kg(-1)) groups, the positive drug Bujiale (0.15 mg x kg(-1)) group, and the sham group. The rats were orally ad-ministrated with drugs for three months. Parathyroid hormone (PTH), procollagen I N-terminal peptide (PINP), alkaline phosphatase (ALP), calcium (Ca) and phosphrous (P) in serum were detected. Femur bones and vertebrae bones of left side were collected to determined bone metrological indexes, including bone mineral density (BMD), bone Ca, and bone ash weight/dry weight percentage. Femur bones of right side were collected to for a morphological observation of bone. RESULT: Compared with the sham group, the model group showed significantly higher PTH and ALP content but obviously lower PINP and Ca content. The three YYH-C 3 groups could resist the decrease of PINP. Specifically, low and middle dose groups could remarkably inhibit the increase of PTH, and the high dose group could increase the Ca content in serum, but without significant effect on the rise in ALP. There was no significant difference in P content in serum in each group. BMD, ash weight/dry weight percentage, Ca and P content of the model group were significantly lower than those in the sham group. The high dose YYH-C group could significantly increase BMD. All of the three YYH-C groups could notably increase ash weight/dry weight percentage and Ca, P content in femur bones and vertebrae bones. YYH-C could significantly increase average thickness, area, area percentage of bony trabeculae, cortical bone area percentage of femoral shaft and the number of osteoblasts on the surface of bony trabeculae, and decrease the number of osteoclasts. CONCLUSION: YYH-C can effectively control the bone mass loss of rats with ovariectomy-induced osteoporosis, prevent the changes in bone microstructure, and inhibit bone absorption, so as to resist high turn-over osteoporosis after ovariectomy. [Key words] total flavones of Epimedium leptorrhizum; ovariectomized rat; osteoporosis

Proteomic investigation of signatures for geniposide-induced hepatotoxicity.

Evaluating the safety of traditional medicinal herbs and their major active constituents is critical for their widespread usage. Geniposide, a major active constituent with a defined structure from the traditional medicinal herb Gardenia jasminoides ELLIS fruit, exhibits remarkable anti-inflammatory, antiapoptotic, and antifibrotic properties and has been used in a variety of medical fields, mainly for the treatment of liver diseases. However, geniposide-induced hepatotoxicity and methods for the early detection of hepatotoxicity have yet to be reported. In this study, geniposide-induced hepatotoxicity was investigated. In addition, candidate biomarkers for the earlier detection of geniposide-induced hepatotoxicity were identified using a label-free quantitative proteomics approach on a geniposide overdose-induced liver injury in a rat model. Using an accurate intensity-based, absolute quantification (iBAQ)-based, one-step discovery and verification approach, a candidate biomarker panel was easily obtained from individual samples in response to different conditions. To determine the biomarkers' early detection abilities, five candidate biomarkers were selected and tested using enzyme-linked immunosorbent assays (ELISAs). Two biomarkers, glycine N-methyltransferase (GNMT) and glycogen phosphorylase (PYGL), were found to indicate hepatic injuries significantly earlier than the current gold standard liver biomarker. This study provides a first insight into geniposide-induced hepatotoxicity in a rat model and describes a method for the earlier detection of this hepatotoxicity, facilitating the efficient monitoring of drug-induced hepatotoxicity.

An investigation of the anti-diabetic effects of an extract from Cladonia humilis.

The effect of Cladonia humilis on glycaemic metabolism was researched in this studied. The blood glucose, insulin secretion and glycogen synthesis of the hyperglycemic mice induced by alloxan were analyzed respectively. The gluconeogenesis and the sugar tolerance of the normal mice were also analyzed in this paper. After the hyperglycemic mice were orally administrated with Cladonia humilis extract, the blood glucose was decreased (p<0.05), the level of insulin secretion and glycogen synthesis were elevated (p<0.05, p<0.01, respectively). In addition, Cladonia humilis extract could inhibit the gluconeogenesis (p<0.01) and improve the sugar tolerance in normal control mice. These results maybe account for the causes of Cladonia humilis extract-induced significant decreases of the blood glucose in hyperglycemic mice.

[Development of gastric precancerous lesion animal model].

OBJECTIVE: To establish a model of gastric precancerous lesion by using Aristolochic manshuriensis which contains aristolochic acids. METHOD: The SD rats were randomly divided into four groups: control and three different doses of ethanol extractive of A. manshuriensis (EEA) (corresponding to aristolochic acid I 2.5, 5.0, 10.0 mg x kg(-1)), respectively. EEA was intragastrically given to rats every other day. At the end of the 10th, 15th, 20th week, part of the rats in each group was sacrificed and the stomachs were weighed. The gastric tumor was assessed by the weight and the relative stomach weight to the body weight. The stomachs were fixed in 4% neutral formalin, and the paraffin imbedding tissues were sliced and HE stained. Histomorphology was observed under the light microscope to determine gastric hyperplasia, mucosa precancerosis (atypical hyperplasia) and gastric cancer formation. RESULT: The rats treated with different doses of EEA for 10 weeks induced mucosa papillary, epithelioma hyperplasia. Histological observation showed mucosa precancerosis lesions characterized as atypical hyperplasia at the dose levels corresponding to aristolochic acid I 5.0 and 10.0 mg x kg(-1) treated for 10 weeks. The incidence rate of gastric precancerosis in those two groups was 100% at the 15th week. Malignant tumors were observed in most of the animals in 10.0 mg x kg(-1) group. The animals in 5.0 mg x kg(-1) group were well tolerant compared to 10.0 mg x kg(-1) group during the course of experiment, so the dose of aristolochic acid I 5.0 mg x kg(-1) and 10-15 weeks treatment were considered to be optimum to establish the model of gastric precancerosis. CONCLUSION: A rat model of gastric precancerosis can be induced within a short duration by giving an oral administration of the ethanol extract of A. manshuriensis which contains aristolochic acids.

[Material and mechanisms for evaluation of Shuanghuanglian injection induced pseudoanaphylactoid reactions].

OBJECTIVE: To investigate the substance basis and the mechanism of pseudoanaphylactoid reactions (PR) induced by Shuanghuanglian injection (SHLI). METHOD: (1)The study of PR and the substance basis of PR of SHLI: ICR mice were divided into different test groups, the mice were intravenously injected with solutions of different concentration of SHLI, baicalin, forsythin, caffeotannic acid, positive control Compound 48/80 and normal sodium. All test substances were mixed with 0.4% Evans blue. The reaction and vascular permeability of the ears were observed and measured 30 min after SHLI injection. (2) The study of mechanisms: Mice were pretreated with an oral administration of Astemizol, intraperitoneal injection of cyclophosphamide 75 mg x kg(-1) or Compound 48/80 4 mg x kg(-1), then mice were intravenously injected with SHLI. At last, vascular permeability of the ears in pretreated groups was compared with SHLI treatment alone group. RESULT: SHLI of 300 mg x kg(-1) and 600 mg x kg(-1) caused obvious vascular hyperpermeability, but baicalin, forsythin and caffeotannic didn't cause vascular hyperpermeability in the ears. The Astemizol can decrease the degree of SHLI-induced vascular hyperpermeability of the ears in the mice. After intraperitoneal injected with cyclophosphamide, there was a slight decrease in the degree of SHLI-induced vascular hyperpermeability, but there was no marked changes in the degree of the SHLI-induced vascular hyperpermeability after the mice were pretreated with Compound 48/80. CONCLUSION: SHLI in clinic equivalent dose can cause vascular hyperpermeability. Baicalin, forsythin and caffeotannic may not result in the PR of SHLI. The mechanism of the PR maybe relate to that SHLI stimulates histamine release, the activation of leucocyte maybe take part in the SHLI-induced PR, too. Antihistamine drug can prevent the genesis of PR which induced by SHLI.

[Evaluation of vitro hepatotoxicity of monocrotaline by precision-cut liver slice technique].

OBJECTIVE: To modify the empirical method of precision-cut liver slice technique, and study the hepatotoxicity of monocrotaline by this technique. METHOD: Liver slices were prepared by the domestic shaking slicer. The technique of precision-cut liver slice was established by detecting MTT reduction used as the slice viability under different culture medium, thickness of slices, pH and culture temperature. After monocrotaline and liver slices co-culture for 6, 24 h, the slice viability, enzyme activity of GPT, GOT, LDH, GGT and protein concentration were detected by MTT reduction, enzyme kinetics method and BCA protein assay method, respectively. RESULT: When the thickness of slices was 200 microm and pH of medium was 6.8, culture temperature was 37 degrees C, BPM culture medium, the viability of slices could maintain on a steady level. LDH leakage was significantly increased and protein content was obviously decreased after monocrotaline co-culture for 24 h with final concentration 0.02, 0.1 and 0.5 g x L(-1). No statistically significant difference between control group and monocrotaline 3 dose groups was observed in the slice viability and the content of GPT, GOT, LDH, GGT and protein after monocrotaline co-culture for 6 h. CONCLUSION: The slice viability could retain 24 h in modified BPM medium surroundings; monocrotaline displayed liver toxicity in some degree after co-culture for 24 hours in 0.02, 0.1 and 0.5 g x L(-1) concentration.

[Review on experiment of traditional Chinese medicine treating to osteoporosis].

Searched the articles between 2000 and 2010, found out and summarized the articles with the topic on the experiment and new techniques of traditional Chinese medicine treating to osteoporosis. The preventive and therapeutic effect to osteoporosis by traditional Chinese medicine had been developed in the past 10 years. The study on standardization of experimental drugs, and the mechanism study with modern cell culture techniques should be enhanced.

[Development of animal model for anaphylactoid test of rodent].

OBJECTIVE: To establish a simple and feasible method of anaphylactoid test on awaked small animals for screening and assessing anaphylactoid reaction of traditional Chinese medicine (TCM) injection with different concentration of tween 80. METHOD: Test substances containing 0.4% Evans blue were intravenously injected into mice at volume of 20 mL x kg(-1) or guinea pigs at a volume of 30 mL x kg(-1). The behaviors were observed and the vascular permeability of ears evaluated by the extent of ear blue staining and absorbance of Evans blue extraction of ears were tested at 30 min after injection. RESULT: Tween 80 solution, Yuxingcao injection with tween 80, and Shuanghuanglian powder injection obviously increased vascular permeability of ears characterized as ear blue staining and increased absorbance of the Evans blue extract from ears extracted by acetone saline both in mice and in guinea pigs in a concentration-dependent (in the case of tween 80) or a dose-dependent (Shuanghuanglian) manner. CONCLUSION: Ear vascular permeability test in mice and guinea pigs can be used as animal models to screen and test anaphylactoid reaction induced by injections.

Comparison of hypoglycemic activity of fermented mushroom of Inonotus obliquus rich in vanadium and wild-growing I. obliquus.

The effects of vanadium-enriched and wild Inonotus obliquus were tested on hyperglycemic mice. The vanadium content of the culture medium was 0.6%, reaching a concentration of 3.0 mg/g in the cultured mushroom while in the wild variety is 1/100 of that amount. The toxicity of vanadium at the 3.0 mg/g level is negligible, but its anti-diabetic effects are significantly different to those of the wild variety (p < 0.05). Due to its high bioavailability and low toxicity, vanadium-enriched I. obliquus could be used as a means of vanadium supplementation, with expectation of obtaining higher bioavailability and lower toxicity in animals.

Displacement of bilirubin from albumin in plasma from jaundiced newborns. An in vitro study of purified Chinese herbal constituents and sulfisoxazole.

The aim of the present study was to clarify the in vitro potential of the purified Chinese herbal constituents LZX-A (neferine), QTJ (sinomenine), YHS (tetrahydropalmitine) and SQZG (notoginsenoside R1) to displace the highly bound bilirubin from albumin binding sites in plasma from jaundiced newborn infants. Sulfisoxazole (1.32 mM) was used as a positive control for bilirubin displacement. The displacing potential of the herbal constituents was investigated at assumed therapeutic concentrations and up to 100 times higher. Total (TB) and unbound (UB) bilirubin in plasma were measured by the peroxidase method. Sulfisoxazole increased the UB concentration in plasma by more than 60%. An increased % displacement of bilirubin was found at higher TB levels confirming the presence also of lower affinity binding sites for bilirubin in plasma. None of the purified herbal constituents showed any bilirubin displacing properties and were unaffected by the level of TB in plasma. The combination of sulfisoxazole and the herbal constituents showed no synergistic effect. It is concluded that none of the investigated purified herbal constituents possess any significant potential in vitro to increase the UB concentration in plasma from jaundiced newborn infants.