This study aims to explore the effect of Sini Decoction on Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB) signaling pathway in the mice with allergic asthma(AA). Forty-eight SPF-grade BALB/c mice were randomly assigned into a blank control group, a model group, a dexamethasone group, and high-, medium-, and low-dose Sini Decoction groups, with 8 mice in each group. The sensitization solution made of ovalbumin and aluminum hydroxide powder was injected intraperitoneally in other groups except the blank control group which was injected with an equal volume of normal saline. The solution(or normal saline) was injected three times in total with an interval of 7 days. At the same time of sensitization, external cold stimulation and ice water were administered in a 4 ℃ climate box for 20 min every day. After modeling, the mice in each group were administrated with corresponding drugs by gavage for 3 weeks. At the end of administration, pentobarbital sodium(30 mg·kg~(-1)) was used for anesthesia, and then the samples were collected for the determination of various indexes. The phenol red test was conducted to evaluate tracheal excretion function. The histopathological changes of lung tissue were observed via hematoxylin-eosin(HE) staining. Masson staining was employed to reveal the deposition of blue collagen fibers around bronchi in lung tissue and the area occupied by blue collagen fibers was calculated. Immunofluorescence method was used to measure the expression of bronchial type Ⅰ collagen(Col-Ⅰ) and α-smooth muscle actin(α-SMA). The protein and mRNA levels of TLR4, NF-κB, cysteinyl aspartate specific proteinase-1(caspase-1), and interleukin-13(IL-13) were determined by Western blot and real-time fluorescence quantitative polymerase chain reaction(real-time PCR), respectively. Compared with the model group, Sini Decoction significantly increased the phenol red excretion from trachea, lowered the lung inflammation score, reduced subepithelial collagen deposition, and decreased Col-Ⅰ and α-SMA levels. Furthermore, the decoction down-regulated the protein and mRNA levels of TLR4, NF-κB, caspase-1, and IL-13 in mouse lung tissue. In conclusion, Sini Decoction can improve air remodeling by inhibiting the TLR4/NF-κB signaling pathway.
Mice ; Animals ; NF-kappa B/metabolism* ; Airway Remodeling ; Interleukin-13/pharmacology* ; Toll-Like Receptor 4/genetics* ; Saline Solution/pharmacology* ; Phenolsulfonphthalein/pharmacology* ; Asthma/genetics* ; Signal Transduction ; Mice, Inbred BALB C ; RNA, Messenger ; Caspases

OBJECTIVETo culture rat prostate glandular epithelial cells and study their barrier functions in vitro.

METHODSRat prostate glandular epithelial cells were cultured in vitro. The expression of the tight junction protein claudin-1 was determined by immunohistochemistry, the structure and composition of the epithelial cells observed under the inverted microscope and transmission electron microscope. The transepithelial electrical resistances (TEERs) were monitored with the Millicell system. The permeability of the prostate glandular epithelial cells was assessed by the phenol red leakage test.

RESULTSCompact monolayer cell structures were formed in the prostate glandular epithelial cells cultured in vitro. Immunohistochemistry showed the expression of the tight junction protein claudin-1 and transmission electron microscopy confirmed the formation of tight junctions between the adjacent glandular epithelial cells. The TEERs in the cultured prostate glandular epithelial cells reached the peak of about (201.3 ± 3.5) Ω/cm2 on the 8th day. The phenol red leakage test manifested a decreased permeability of the cell layers with the increase of TEERs.

CONCLUSIONThe structure and function of rat prostate glandular epithelial cells are similar to those of brain capillary endothelial cells, retinal capillary endothelial cells, and intestinal epithelial cells. In vitro cultured prostate glandular epithelial cells have the barrier function and can be used as a model for the study of blood prostate barrier in vitro.

Animals ; Cell Membrane Permeability ; Cells, Cultured ; Claudin-1 ; metabolism ; Electric Impedance ; Epithelial Cells ; pathology ; physiology ; ultrastructure ; In Vitro Techniques ; Male ; Microscopy, Electron, Transmission ; Phenolsulfonphthalein ; pharmacokinetics ; Prostate ; metabolism ; pathology ; Rats ; Tight Junctions

Membrane-bound HLA-G (mHLA-G) discovery on adipose derived stem cells (ADSCs) as a tolerogenic and immunosuppressive molecule was very important. Many documents have shown that HLA-G expression can be controlled via some hormones such as progesterone (P4) and estradiol (E2). Therefore, this study was designed to evaluate progesterone and estradiol effects on mHLA-G in ADSCs at restricted and combination concentrations. Three independent cell lines were cultured in complete free phenol red DMEM and subcultured to achieve suffi cient cells. These cells were treated with P4, E2 and P4 plus E2 at physiologic and pregnancy concentrations for 3 days in cell culture conditions. The HLA-G positive ADSCs was measured via monoclonal anti HLA-G-FITC/MEMG-09 by means of flow cytometry in nine groups. Data were analyzed by one way ANOVA and Tukey's post hoc tests. There were no signifi cant values of the mean percentage of HLA-G positive cells in E2-treated and the combination of P4 plus E2-treated ADSCs compared to control cells (p value>0.05) but P4 had a signifi cant increase on mHLA-G in ADSCs (p value<0.05). High P4 concentration increased mHLA-G but E2 and the combination of P4 plus E2 could not change mHLA-G on ADSCs.
Cell Culture Techniques ; Cell Line ; Estradiol* ; Flow Cytometry ; HLA-G Antigens* ; Phenolsulfonphthalein ; Pregnancy ; Progesterone* ; Stem Cells*

OBJECTIVETo observe the efficacy and the influence on quality of life (QOL) of syndrome differentiation treatment with Chinese medicine (CM) for opioid-induced constipation as well as the safety and influence on analgesic effect of opioids.

METHODSTotally 406 cases enrolled from 53 collaborating medical centers were randomly assigned to a CM group and a control group. The CM group were treated with CM decoction based on syndrome differentiation, and the control group were treated with Phenolphthalein Tablet. Both groups were treated for 14 days. Cleveland constipation score (CCS), numerical rating scale (NRS) of pain and Chinese version of European Organisation for Research and Treatment of Cancer, Quality of Life Questionnaire-C30 V3.0 (EORTC QLQ-C30 V3.0) were used to evaluate the efficacy, pain controlled and QOL status.

RESULTSThe comparisons of CCS score reduction and QOL between the two groups after treatment suggested that the improvements of constipation and QOL in the CM group were better than that in the control group (P<0.05). The total efficiency of the CM group was better than the control group (93.5% vs. 86.4%, P<0.05). There was no significant difference in NRS scores between before and after treatment in both groups. There was no serious drug-related adverse event during the course of study.

CONCLUSIONCM decoction could effectively treat opioid-induced constipation and improve patients' QOL at the same time. It is safe and doesn't affect the analgesic effect of opioids when treating constipation.

Aged ; Analgesics, Opioid ; administration & dosage ; adverse effects ; Constipation ; chemically induced ; drug therapy ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neoplasms ; complications ; Pain, Intractable ; drug therapy ; Phenolphthalein ; administration & dosage ; Quality of Life ; Treatment Outcome

OBJECTIVETo study the analgesic, expectorant and antitussive effects of the compatible use of Aconiti Radix Cocta and Fritillaria cirrhosa or F. thunbergii with different matching ratio or dose in mice.

METHODThe two-factor, seven-level uniform design method was adopted to observe the analgesic, expectorant and antitussive effects of the oral administration with the two combined decoctions in rats, with frequency of body torsions induced by acetum, secretion of phenol red in tracheas and frequency of coughs as indexes. Significant matching proportions and doses were collected for verification.

RESULTThe effect on the frequency of body torsions: The combined decoctions could effectively reduce the frequency of body torsions. According to a regression analysis, Aconiti Radix Cocta and F. cirrhosa had the synergistic effect, which was maximized with a ratio of 1: 1. The 1: 1 combined decoction played the least role in reducing the frequency of body torsions with a total dose of more than 5 g x kg(-1). The effect on the secretion of phenol red in tracheas. The combined decoctions could effectively increase the secretion of phenol red in tracheas. According to a regression analysis, Aconiti Radix Cocta and F. thunbergii had the antagonism, which was maximized at the ratio of 1: 1, and minimized with a total dose of less than 10 g x kg(-1) and a ratio of 5: 1 between F. thunbergii and Aconiti Radix Cocta. The effect on the frequency of coughs. The combined decoctions could effectively reduce the frequency of coughs. According to a regression analysis, Aconiti Radix Cocta and F. cirrhosa had the antagonism, which was maximized at the ratio of more than 1: 5 and less than 10: 1. There was no interaction between Aconiti Radix Cocta and F. thunbergii. F. thunbergii could reduce the frequency of coughs, whereas Aconiti Radix Cocta showed no effect.

CONCLUSIONThe compatible application of Aconiti Radix Cocta and F. cirrhosa could enhance the analgesic effect of Aconiti Radix Cocta and reduce the expectorant and antitussive effects of F. cirrhosa, which vary according to different matching ratio and dose. The compatible application of Aconiti Radix Cocta and F. thunbergii shows no effect on the antitussive effect of F. thunbergii. This study provides experimental basis for in-depth studies on the combined effect of Aconiti Radix Cocta and Fritillaria--two of eighteen incompatible pairs.

Aconitum ; chemistry ; Analgesics ; pharmacology ; Animals ; Antitussive Agents ; pharmacology ; Behavior, Animal ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Expectorants ; pharmacology ; Fritillaria ; chemistry ; Male ; Mice ; Phenolsulfonphthalein ; metabolism ; Trachea ; drug effects ; metabolism

OBJECTIVETo develop a method for simultaneous assay of propulsion and absorption in small intestine.

METHODSThe mice were administrated through gastric tube with mixed reagents containing 0.12% phenol red, D-xylose (1.25%, 2.5% and 5%) and 15% gelatin. The influence of phenol red on D-xylose absorption and the influence of D-xylose on small intestine propulsion rate were investigated by measuring serum concentration of D-xylose with phloroglucinol method.

RESULTSAt 10 min, no significant difference was found between 5% D-xylose mixed reagent group and 5% D-xylose control. At 15 min, small intestine propulsion rate in 5% D-xylose mixed reagent group, but not in 2.5% and 1.25% D-xylose mixed reagent groups, was significantly higher than in phenol red control (P<0.05).

CONCLUSIONGastric administration of mixed reagent containing 0.12% phenol red, 5% D-xylose and 15% gelatin can simultaneously assay propulsion and absorption of small intestine in mice.

Animals ; Biological Assay ; methods ; Intestinal Absorption ; Intestine, Small ; metabolism ; physiology ; Male ; Mice ; Mice, Inbred ICR ; Peristalsis ; Phenolsulfonphthalein ; pharmacokinetics ; Xylose ; pharmacokinetics

PURPOSE: To evaluate the clinical usefulness of the phenol red thread test as a diagnostic tool of dry eye by comparing the phenol red thread test, Schirmer's test and tear break-up time. METHODS: The present study included 30 dry eye patients belonging to dry eye workshop grade 1 or 2 and 25 normal subjects. Phenol red thread test, Schirmer's test, and tear break-up time were performed on each subject's right eye. The sensitivity, specificity and repeatability of each test were compared, and the correlations between the 3 tests were also analyzed. RESULTS: Tear break-up time was superior to the other tests in terms of sensitivity and repeatability. The phenol red thread test was better than Schirmer's test in terms of specificity and repeatability. In all 55 patients including dry eye patients and normal subjects, the phenol red thread test showed a greater correlation with tear break-up time than did Schirmer's test. In addition, in 25 dry eye patients, the correlation between the phenol red thread test and Schirmer's test increased significantly. CONCLUSIONS: The phenol red thread test is less irritating and requires a shorter testing time than Schirmer's test. Additionally, the phenol red thread test is superior to Schirmer's test in terms of specificity, repeatability, and relation to tear break-up time. In addition, the correlation between the phenol red thread test and Schirmer's test significantly increases in dry eye patients. Therefore, the phenol red thread test is a good substitute option for Schirmer's test in diagnosing dry eye.
Eye ; Humans ; Phenol ; Phenolsulfonphthalein ; Sensitivity and Specificity ; Tears

To determine the optimal media conditions for the detection of the extracellular cellulase activity in Ganoderma neo-japonicum, we varied three media conditions: dye reagent, pH, and temperature. We evaluated the use of four dyes, Congo red, phenol red, remazol brilliant blue, and trypan blue. To observe the effect of pH on the chromogenic reaction, we tested media ranging from 4.5 to 8.0. To research the effect of temperature on the clear zone and the fungus growing zone, we tested temperatures ranging from 15 to 35degrees C. On the whole, the best protocol called for Ganoderma neo-japonicum transfer onto media containing Congo red with a pH of 7.0, followed by incubation at 25degrees C for 5 days. Our results will be useful to researchers who study extracellular enzyme activity in Ganoderma neo-japonicum.
Benzenesulfonates ; Cellulase ; Coloring Agents ; Congo Red ; Diminazene ; Fungi ; Ganoderma ; Hydrogen-Ion Concentration ; Phenolsulfonphthalein ; Trypan Blue

To obtain basic information on the detection of cellulolytic activity in Auricularia auricula-judae, the influences of dye reagent, pH, and temperature were assessed. Chromogenic dye (congo red, phenol red, remazol brilliant blue, and trypan blue) was individually incorporated into a medium containing either carboxymethyl-cellulose, Avicel, or D-cellobiose as a polysaccharide carbon substrate. The other assessments utilized pHs ranging from 4.5 to 8.0 and temperatures from 15~35degrees C. Overall, when A. auricula-judae species were transferred onto media contained Congo red and adjusted pH 7.0 and then incubated at 25degrees C for 5 days, the clear zone indicative of cellulolytic activity was more pronounced.
Benzenesulfonates ; Carbon ; Cellulose ; Congo Red ; Diminazene ; Hydrogen-Ion Concentration ; Phenolsulfonphthalein

To determine the optimal medium conditions for the detection of the cellulolytic activity in Ganoderma lucidum, we varied three media conditions: dye reagent, pH, and temperature. First, we evaluated the use of four dyes, Congo Red, Phenol Red, Remazol Brilliant Blue, and Trypan Blue. To observe the effect of pH on the chromogenic reaction, we also made and tested various media spanning acidic and alkaline pHs, ranging from 4.5 to 8.0. Furthermore, in order to research the effect of temperature on the clear zone and the fungus growing zone, we tested temperatures ranging from 15 to 35degrees C. On the whole, the best protocol called for Ganoderma lucidum transfer onto media containing Congo red with pH adjusted to 7.0, followed by incubation at 25degrees C for 5 days. Our results will be useful to researchers who aim to study extracellular enzyme activity in Ganoderma lucidum.
Benzenesulfonates ; Coloring Agents ; Congo Red ; Diminazene ; Fungi ; Ganoderma ; Hydrogen-Ion Concentration ; Phenolsulfonphthalein ; Reishi ; Trypan Blue