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1.
Article in Chinese | WPRIM | ID: wpr-887969

ABSTRACT

Polyethylene glycol (PEG) surface film-forming method was used to prepare hydrophilic Indigo Naturalis decoction pieces with stable effect.The preparation process of modified Indigo Naturalis was optimized and its microscopic properties,hydrophilicity,antipyretic efficacy,and safety were systematically evaluated.With equilibrium contact angle as assessment index,the influence of modifier type,modifier dosage,dispersant dosage,and co-grinding time on water solubility of Indigo Naturalis was investigated by single factor test.The results showed that the optimal preparation process was as follows.The 6%PEG6000 is dissolved in 10%anhydrous ethanol solution by sonification and then the mixture is ground with Indigo Naturalis for 2 min.The resultant product is dried on a square tray in an oven at 60℃to remove ethanol and thereby the PEG-modified hydrophilic Indigo Naturalis decoction pieces are yielded.The morphological observation under scanning electron microscope (SEM) indicated that the modified Indigo Naturalis had smoother surface than Indigo Naturalis,and energy spectrometer measurement showed that the nitrogen (N),calcium(Ca),oxygen (O),and silicon (Si) on the surface of modified Indigo Naturalis powder were less than those of Indigo Naturalis powder.Modified Indigo Naturalis had the equilibrium contact angle 18.96°smaller,polar component 22.222 m J·m~(-2)more,and nonpolar component 7.277 m J·m~(-2)smaller than the Indigo Naturalis powder.Multiple light scattering technique was employed to evaluate the dispersion in water and the result demonstrated that the transmittance of Indigo Naturalis and modified Indigo Naturalis was about85%and 75%,respectively,suggesting the higher dispersity of modified Indigo Naturalis.The suspension rate of modified Indigo Naturalis in water was determined by reflux treatment.The result showed that 57%of Indigo Naturalis was not wetted after refluxing for1 h,while the modified Indigo Naturalis was all wetted and dispersed into water.The dissolution of indigo and indirubin of modified Indigo Naturalis increased and the process was more stable.Then,rats were randomized into the blank group,model group,acetaminophen group,Indigo Naturalis group,and hydrophilic Indigo Naturalis group.The temperature changes of rats were observed after administration and the concentration of IL-1βand TNF-αin serum and IL-1βand PGE_2in hypothalamus was measured.The results indicated that the temperature of Indigo Naturalis group and hydrophilic Indigo Naturalis group dropped and the IL-1βlevel of the hydrophilic Indigo Naturalis group decreased (P<0.05) as compared with those in the model group.Thus,both Indigo Naturalis and hydrophilic Indigo Naturalis had antipyretic effect,particularly the hydrophilic Indigo Naturalis.The acute toxicity test of hydrophilic Indigo Naturalis verified that it had no toxicity to rats.In this study,the hydrophilic Indigo Naturalis decoction pieces were prepared with the PEG surface film-forming method,and the antipyretic efficacy and safety were evaluated,which expanded the technological means of powder modification for Chinese medicine and provided a method for clinical use of Chinese medicine.


Subject(s)
Animals , Drugs, Chinese Herbal , Hydrophobic and Hydrophilic Interactions , Indigo Carmine , Indigofera , Polyethylene Glycols , Rats
2.
Article in Chinese | WPRIM | ID: wpr-887968

ABSTRACT

As an effective antipyretic medicine,Indigo Naturalis has a long history of application in the field of Chinese medicine.The content of organics,mainly indigo and indirubin,is about 10%. However,the active ingredients and mechanism of its antipyretic effect have not yet been fully elucidated. In view of this,they were investigated in this study with the rectal temperature change as an indicator and 2,4-dinitrophenol-induced fever rats as subjects. The content of PGE2 and c AMP in the hypothalamus and the serum levels of TNF-α,IL-1β and IL-6 were determined by ELISA. Moreover,the plasma samples of fever rats were analyzed by metabonomics in combination with UPLC-Q-TOF-MS for the exploration of potential biomarkers and the discussion on the antipyretic mechanism of Indigo Naturalis and its active ingredients. The results showed that the rising trend of rectal temperature in rats was suppressed 0. 5 h after the treatment with Indigo Naturalis,organic matter,indigo or indirubin as compared with the rats of model group( P < 0. 05),among which Indigo Naturalis and organic matter had better antipyretic effect. ELISA results showed that organic matter and indigo can inhibit the expression of PGE2 and c AMP( P<0. 01),while Indigo Naturalis and organic matter were effective in curbing the increase in TNF-α( P<0. 05). A total of 21 endogenous metabolites were identified from the plasma samples of the Indigo Naturalis,organic matter,indigo and indirubin groups,which were mainly involved in glycerophospholipid metabolism.


Subject(s)
2,4-Dinitrophenol , Animals , Antipyretics , Drugs, Chinese Herbal , Indigo Carmine , Indigofera , Rats
3.
Article in Chinese | WPRIM | ID: wpr-887967

ABSTRACT

Indigo Naturalis( IN) is mainly composed of 10% organic matter and 90% inorganic matter,with a poor wettability and strong hydrophobicity. Indigo,indirubin and effective ingredients are almost insoluble in water. And how it exerts its effect after oral administration still needs to be revealed. For this reason,this study put forward the hypothesis that " Indigo Naturalis forms a slightly soluble calcium carbonate carrier in a strong acid environment of gastric fluid,and organic substances are solubilized in the bile environment of intestinal fluid",and then verified the hypothesis. First,the dissolution apparatus was used to simulate the change process of IN in different digestive fluid,and the effects of low-dose and normal bile on the dissolution of inorganic substances and the release of organic substances were compared. After the surface morphology and element changes of IN in different digestive fluid were observed,it was found that bile is the key to promoting the dissolution of organic and inorganic substances in IN. Furthermore,the rat fever model induced by 2,4-dinitrophenol was used to study the antipyretic effect of IN in normal rats and bile duct ligation rats. It was found that the antipyretic effect of IN on normal rats was better than that of bile duct ligation rats. The above results indicated that after oral administration of IN,the calcium carbonate carrier was transformed into a slightly soluble state in acidic gastric fluid,and a small amount of organic matter was released. When IN entered the intestinal fluid mixed with bile,the carrier dissolved in a large amount,and indigo and indirubin were dissolved in a large amount,so as to absorb the blood and exert the effect. This study has a certain significance for guiding clinical application of IN. For patients with insufficient bile secretion( such as bile duct resection),oral administration with IN may not be effective and shall be paid attention.


Subject(s)
Animals , Bile , Humans , Hydrophobic and Hydrophilic Interactions , Indigo Carmine , Indigofera , Plant Extracts , Rats
4.
Article in Chinese | WPRIM | ID: wpr-636241

ABSTRACT

Objective To investigate the imaging features of active Crohn′s disease on conventional ultrasound and contrast-enhanced ultrasound (CEUS). Methods The imaging features of 20 patients with an established diagnosis of Crohn′s disease on transabdominal high-frequency ultrasound and contrast-enhanced ultrasound in Shanghai Tenth People′s Hospital from August 2011 to December 2012 were studied retrospectively. Contrast-specific imaging modes were performed and the ultrasound contrast agent was SonoVue. The thickness of inner, outer and all layers of intestinal walls in the lesion area were observed;the ratio between inner and outer bowel wall thickness was calculated;Limberg classiifcation was determined by Power-Doppler results. Likewise, contrast-enhanced ultrasound was used to evaluate the degree and area of bowel wall enhancement, as well as the changes over time. Variance analysis was applied to compare intestinal wall thickness, arrive time of contrast agent, time to peak and washing time of patients with Crohn′s disease from different Limberg groups, and further comparison between groups were anlysed with LSD-t test. Results The intestinal wall thickness of all 20 patients was larger than 4 mm, while the mean thickness of intestinal walls was (8.8±0.4) mm (range 5.5-12.0 mm);the ratio between inner and outer wall thickness was greater than 1.0. Limberg classiifcation wasⅡin 2 patients,Ⅲin 8 patients andⅣin 10 patients. There were two enhancement patterns shown on contrast-enhanced ultrasound:Pattern 1 in 13 (13/20, 65.0%) patients showing simultaneous enhancement in both inner and outer intestinal walls at the same time. Pattern 2 in 7 (7/20, 35.0%) patients showing outward enhancement from inner to outer wall with a predominance of inner wall. The wall thicknesses of patients with Crohn′s disease from Limberg Ⅱgroup, Limberg Ⅲgroup and Limberg Ⅳgroup were (6.6±0.1), (7.5±0.4) and (10.2±0.4) mm respectively. The thicknesses of inner bowel walls were (3.6±0.6), (5.0±0.2) and (7.3±0.3) mm respectively. CEUS time to peak was (30.5±2.1), (26.9±2.4) and (21.0±1.6) s respectively. The wash-in time of the contrast agent was (18.0±5.7), (10.6±1.0) and (8.7±1.2) s respectively. As the Limberg level increased, the thickness of the entire and inner bowel wall both increased, while CEUS time to peak and wash-in time of the contrast agent became longer. These difference was statistically significant. In addition, the ratio between inner and outter wall thickness also increased as the Limberg level increased, however, the difference was statistically insigniifcant. Likewise, the outer bowel wall thickness and the arrival time of the contrast agent in patients with Crohn′s diseases from different Limberg level groups showed no statistical significance. Conclusions The patients with active Crohn′s disease always showed thickened bowel walls, higher Limberg level and complete or partial enhancement of bowel wall on CEUS. There were some correlations between the above-mentioned ifndings.

5.
Chinese Medical Journal ; (24): 898-903, 2008.
Article in English | WPRIM | ID: wpr-258569

ABSTRACT

<p><b>BACKGROUND</b>Patients with sliding hiatus hernia (HH) and reflux esophagitis (RE) usually suffer from esophageal dysmotility. The aim of the present study was to investigate the role of acid reflux and duodenal gastroesophageal reflux (DGER), esophageal manometry, and esophageal dysmotility by applying the barium meal examination.</p><p><b>METHODS</b>RE with HH was initially diagnosed using the reflux disease questionnaire, and was further confirmed by a barium meal examination and an endoscopy. The radiographic technique was used to test for spasms, strictures, and the coarseness of the mucosa, also was to study the types of reflux and clearance. Then, the esophageal manometry, the esophageal 24-hour pH, and the bilirubin monitoring were observed.</p><p><b>RESULTS</b>Fifty-five patients were diagnosed as HH combined with RE and divided into two groups according to the severity of their esophagitis: group HH1 (grades A and B) and group HH2 (grades C and D). The barium meal examination revealed that the mucosa was either granular or nodular in all cases. The dump reflux and delayed clearance were more significant in patients in the HH2 group than those in the HH1 group (P < 0.05). The percentages of total, supine, and upright acid exposure time were greater in patients with HH than those in the control group (P < 0.01), but the differences between the HH1 and the HH2 groups were not significant. Lower esophageal sphincter pressure (LESP) was lower in the HH group than in the control group (P < 0.05). Three DGER parameters: the percentage of time with absorbance greater than 0.14, the number of bile reflux episodes, the number of bile refluxes lasting longer than 5 minutes were (28.43 +/- 23.34), (40.57 +/- 31.30), and (15.15 +/- 8.72), respectively in the HH2 group; these statistics were significantly higher than those for the HH1 (P < 0.05). The frequency and amplitude of peristalsis were all lower in HH patients than in the control (P < 0.05). Of all the patients, 54.3% (30 of 55) with acid reflux and DGER simultaneously in the HH group exhibited refluxes of barium from the stomach to the esophagus in the recumbent position, and 29.4% (5 in 17) with delayed clearance in the HH group were correlated with esophageal body peristalses. The result was that the frequency and amplitude of peristalsis were less and the duration of esophageal peristalsis was longer than those of control group.</p><p><b>CONCLUSIONS</b>Esophageal dysmotility may play an important role in the severity of RE combined with HH. Esophageal motility results on a barium examination may coincide with esophageal manometry, 24-hour pH, and bilirubin monitoring in the RE and HH, but the radiologic method was the simplest to apply.</p>


Subject(s)
Adult , Aged , Bilirubin , Esophageal Motility Disorders , Pathology , Esophageal pH Monitoring , Esophagitis, Peptic , Pathology , Esophagoscopy , Female , Hernia, Hiatal , Pathology , Humans , Male , Manometry , Methods , Middle Aged
6.
Article in Chinese | WPRIM | ID: wpr-268062

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the differential expression of isocitrate lyase in Penicillium marneffei phagocytized by nonstimulated and stimulated murine macrophages, and explore the role of glyoxylate pathway in pathogenesis of Penicilliosis marneffei.</p><p><b>METHODS</b>Penicillium marneffei conidia and Raw264.7 cells were incubated in 16 cultures, which were divided to 4 groups for treatment with N-monomethyl-L-arginine (LNMMA, CI group), murine interferon-gamma (IFN-gamma) plus lipopolysaccharide (LPS) (T group), IFN-gamma plus LPS and LNMMA (TI group), or the same volume of culture medium (C group). The transcriptional levels of isocitrate lyase were detected using real-time RT-PCR, and its expression levels detected biochemically.</p><p><b>RESULTS</b>The transcriptional levels of isocitrate lyase in C, CI, T, TI groups were 1.00, 1.42, 33.09, and 74.88 (P<0.05), while the expression levels were 0.06, 0.07, 0.18, and 0.93, respectively (P<0.05). The content of nitric oxide in T group was significantly higher than that in the other groups (P<0.01), but the CFU of T group was the lowest (P<0.01).</p><p><b>CONCLUSION</b>Reactive nitrogen intermediates induced by stimulated murine macrophages restrain the expression of isocitrate lyase of Penicillium marneffei and development of Penicillium marneffei, in which process the glyoxylate pathway may play an important role.</p>


Subject(s)
Animals , Cell Line , Fungal Proteins , Genetics , Gene Expression Profiling , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Fungal , Host-Pathogen Interactions , Interferon-gamma , Pharmacology , Isocitrate Lyase , Genetics , Lipopolysaccharides , Pharmacology , Macrophages , Allergy and Immunology , Microbiology , Mice , Nitric Oxide , Allergy and Immunology , Penicillium , Genetics , Allergy and Immunology , Physiology , Phagocytosis , Allergy and Immunology , Reverse Transcriptase Polymerase Chain Reaction , omega-N-Methylarginine , Pharmacology
7.
Article in Chinese | WPRIM | ID: wpr-353216

ABSTRACT

<p><b>OBJECTIVE</b>To clone luffin-a cDNA from the seeds of Luffa cylindrical, and to obtain bioactive recombinant luffin-a protein using the expression vector pET-44a (+) in E. coli.</p><p><b>METHODS</b>The cDNA sequence encoding luffin-a was cloned from the fresh seeds of Luffa cylindrical by RT-PCR. The target DNA fragments were sequenced after T-A cloning. The luffin-a expression plasmid was constructed by inserting the luffin-a cDNA fragment into vector pET-44a (+). Luffin-a was expressed in E. coli by addition of IPTG into final concentration 1.0 mmol/L. The recombinant luffin-a was identified by SDS-PAGE. The biological activity of luffin-a protein was evaluated by using the MTT assay in HepG2 cells following fluid-phase endocytosis.</p><p><b>RESULTS</b>In comparison with the reported luffin-a, the homology of nucleotide sequence of the cloned luffin-a gene was 99.73%, while their amino acid sequences were identical. The solubility of recombinant protein was analyzed by SDS-PAGE, and the luffin-a was mainly produced in inclusion bodies. The recombinant luffin-a, renatured by dialysis of the denatured products, showed a similar cytotoxicity to ricin A chain.</p><p><b>CONCLUSION</b>The cDNA of luffin-a has been successfully cloned. The recombinant luffin-a protein expressed by E. coli is bioactive.</p>


Subject(s)
Amino Acid Sequence , Base Sequence , Cloning, Molecular , Luffa , Chemistry , Molecular Sequence Data , Plant Proteins , Genetics , Polymerase Chain Reaction , Ribosome Inactivating Proteins, Type 1 , Seeds , Chemistry
8.
Article in English | WPRIM | ID: wpr-254012

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the potential role of preoperative adjuvant chemotherapy on early stage cervical squamous carcinoma with bulky tumor.</p><p><b>METHODS</b>One hundred and forty-five patients with cervical squamous cancer stages Ib-IIa were investigated, among which 17 patients with bulky tumors (> or = 4 cm) were managed by cisplatin-based chemotherapy for 1-2 courses followed by radical hysterectomy and pelvic lymphadenectomy (BC group). The change of tumor size, pelvic lymph nodes metastasis, cervical wall invasion, the involvement of surgical specimen margin, and the blood loss during operation were assessed after operation and compared with those in 51 patients with bulky tumors (BN group) and 77 patients with small local tumors (S group) who underwent surgery directly.</p><p><b>RESULTS</b>(1) The tumor size of 17 patients in BC group were decreased in various degrees after chemotherapy, with 13 patients of clinical effectiveness (76.47%). And the responsiveness pertained to neither histological differentiation nor size of local tumors. (2) Post-operative histology has showed that patients in BC and BN group have higher incidence of lymph node metastasis and deep cervical infiltration (5/68 and 3/68, respectively) than in S group (1/77 and 1/77, respectively) while with no statistical significance. (3) Blood loss during operation in BC group was less than BN and S group. (4) Seventeen patients, including those underwent surgeries of vaginal prolongation and/or ovarian transposition, appeared disease-free survival within the follow-up time.</p><p><b>CONCLUSIONS</b>Most of patients with bulky early stage cervical squamous carcinoma are sensitive to cisplatin-based chemotherapy, which could greatly reduce local tumor size and in turn facilitate the following operation by well controlling blood loss.</p>


Subject(s)
Adult , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Bleomycin , Carcinoma, Squamous Cell , Drug Therapy , Pathology , General Surgery , Chemotherapy, Adjuvant , Cisplatin , Drug Administration Schedule , Etoposide , Female , Humans , Hysterectomy , Lymphatic Metastasis , Middle Aged , Neoplasm Staging , Preoperative Care , Uterine Cervical Neoplasms , Drug Therapy , Pathology , General Surgery , Vincristine
9.
Article in Chinese | WPRIM | ID: wpr-682637

ABSTRACT

Objective To investigate esophageal response to intraesophageal balloon-distention and acid perfusion stimuli and to evaluate the visceral hypersensitivity in non erosive reflux disease (NERD) patients.Methods Thirty-one NERD patients diagnosed by reflux disease questionnaire (RDQ) and endoscopy and 10 control subjects were enrolled in this study.Esophageal mechanical sensitivity was measured by esophageal barostat and recorded as initial perception threshold and maximal tolerated pain threshold by volume.The chemical sensitivity was measured by acid perfusion test,and quantified by two parameters (trigger time and acid related symptom score).Results Initial perception threshold and maximally tolerated pain threshold of NERD patients was (9.6?4.8) ml and (12.3?3.2) ml, significantly lower than those of controls,(13.2?7.5) ml and (21.6?5.7) ml,respectively (P

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