Aim To investigate the inhibitory effect of paeonol (Pae) on the human esophageal cancer cell line Eca-109 in vitro and in vivo and its effect on apoptosis.Methods Cytotoxic effect of Pae on Eca-109 cells cultured in vitro was measured by MTT assay.Anti-tumor activity was performed on BALB/c nude mice xenografts model.The morphologic changes of tumor tissue were observed under light microscope and transmission electron microscope.The apoptosis index was assessed by TUNEL.Results Pae had significant in- hibitory effect on the proliferation of Eca-109 cells,and the IC50 value was 0.342 mmol?L-1.In the model of human esophageal cancer xenografts in BALB/c nude mice,the inhibitory rates of Pae group (25、50、100、200 mg?kg-1) were 10.67%、23.54%、27.91% and 34.46% respectively.In vivo administration of Pae 100 mg?kg-1 combined with cisplatin 5 mg?kg-1 resulted in a significant inhibition of Eca-109 tumor growth with the inhibitory rate of 77.91%,compared with cisplatin used alone (58.71%).The more apoptotic tumor cells could be seen under light microscope in every theraperutic groups than those in control.Changes of ultrastructure of tumor cells including concentration and side accumulation of the nuclear chromatin,and the fragmentation of the nuclear was observed under transmission electronic microscope.Apoptosis body was also found.The apoptosis indexes of every theraperutic groups were significantly different from the control.Conclusion Pae can inhibit the cell growth and induce apoptosis in human esophageal cancer cell line Eca-109 in vitro and in vivo.

ObjectiveTo explore the effect of Shuanglong pill which can vivid blood and relax thread pulse on cardial morphology and haemodynamics in experimental myocardial infarction (MI) rats.Methods66 rats ligated left coronary artery were randomly divided into large dose group of Shuanglong pill (LDG), small dose group of Shuanglong pill (SDG), the normal control (NC) and the control group of MI. Half in each group received 2 week treatment and the other received 4 week treatment, and were spiled in femoral and common carotid artery respectively. Parameters of cardiac haemodynamics were monitored by eight lead physiological recorder. ResultsLeft ventricular systolic maximum velocity in LDG, SDG and DC was more significantly elevated than in MI, after 2 weeks and 4 weeks of treatment, but it was lower than NC. There were no significant differences among LDG, SDG and DC. Ventricular systolic maximum velocity in LDG was more significantly elevated than in MI after 2 week treatment.ConclusionTreating MI with Shuanglong pill can improve left ventricular systolic maximum velocity, and diastolic phase function of left ventricular also elevate during the acute stage of MI by using large dose.

Cerebral Infarction ; blood ; Humans ; Interleukin-10 ; blood ; Interleukin-17 ; blood

Child, Preschool ; Female ; Humans ; Infant ; Interleukin-1beta ; blood ; cerebrospinal fluid ; Male ; Seizures, Febrile ; blood ; cerebrospinal fluid ; Zinc ; blood ; cerebrospinal fluid

Based on the Bacillus sp., isolated from Lop Nur Desert, the technology of separation and purification and the antioxidant effect were studied. After centrifugation and vacuum filtration, the deproteinization of supernatant was operated with Sevag reagent. The crude exopolysaccharide (EPS) was obtained by precipitation with ethanol. The optimum conditions for the isolation were as follow: pH 7.0, temperature 4?C, time 1.5 h, and material to ethanol ratio 1: 4. Dissolved in water, the crude EPS was fractional separated on activated carbon column (1.5 cm ? 24 cm), eluted with distilled water, 60% ethanol, 95% ethanol, and the main fraction was collected. Then the EPS was purified on Sephadex G-100 gel column, eluted with NaCl (0.2 mol/L). Fractions (4 mL, each) were also combined according to total sugar by phenol-sulfuric acid method and protein content was determined by Coomassie brilliant blue. The results showed that EPS was relatively homogeneous glycoprotein. The data of antioxidation in vitro showed that the EPS had a high antioxidant activity, which could quench hydroxyl radical, superoxide radical and had antilipid peroxidation activity. All of these indicated that EPS was a good natural antioxidant.

The cases of lymphoma accompanied or preceded by Coombs' test positive autoimmune hemolytic anemia (AIHA) have been reported. However, Coombs' test negative AIHA prior to the diagnosis of lymphoma was rarely described. Herein, this article reports a case of non-Hodgkin's lymphoma (NHL) preceded about 1.5 years by Coombs test negative AIHA. A woman aged 69 was diagnosed with HA based on the history and laboratory tests. Further studies revealed that this patient was negative with Coombs' test for IgG, IgM, IgA and C3. After all possible causes of HA, especially malignancies were ruled out, the patient was diagnosed with Coombs' test negative AIHA and treated with prednisolone. The patient responded well initially to steroid treatment. Two recurrences of acute HA were presented at time of 10 months post steroid cessation, and immediately after an attempt to withdraw steroid, respectively, but the hemolysis was effectively controlled by reinstitution of prednisolone. At third recurrence, however, the patient was no longer responding to steroid, and was found with cervical lymphadenopathy. Coombs' test for IgG, IgM, IgA and C3 remained negative. B cell NHL was diagnosed by pathology. After receiving 6 cycles of CHOP chemotherapy, the patient was lymphoma free, but the hemolysis was not improved, however, which was effectively controlled by the following low dose-rituximab (RTX) therapy. The patient was still kept in a remission of lymphoma free of anemia. In conclusion, this report presented a very rare case of NHL with Coombs' test negative AIHA as initial major clinical manifestation.
Aged ; Anemia, Hemolytic, Autoimmune ; diagnosis ; etiology ; therapy ; Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Coombs Test ; Female ; Humans ; Lymphoma, Non-Hodgkin ; complications ; diagnosis ; Rituximab

OBJECTIVETo investigate the method for culturing corpus cavernosum smooth muscle cells (CCSMs) derived from diabetic rats with erectile dysfunction (ED) for the study of ED caused by diabetes.

METHODSCCSMs were isolated from the corpus cavernosum of diabetic rats with ED and cultured using a modified method of adherent tissue culture. The cultured cells were identified by immunohistochemistry and the cell morphology and proliferation were observed.

RESULTSThe primary culture of CCSM was performed successfully, and the cells were seen to migrate from the small tissue pieces 3 days later, reaching nearly confluence in 16-18 days. A typical "hill-valley" growth pattern was noted in the cell passaging. Immunohistochemical staining for alpha-smooth muscle actin (alpha-SM-actin) and desmin yielded positive results in the cells.

CONCLUSIONThe modified method for adherent tissue culture is convenient and reliable in establishing the in vitro cell culture model of CCSMs from diabetic rats with ED, and the cultured CCSMs display a faster proliferation than normal CCSMs. No obvious differences in the cell morphology can be found between diabetic and normal CCSMs under light microscope.

Animals ; Cell Culture Techniques ; Cells, Cultured ; Diabetes Mellitus, Experimental ; complications ; pathology ; physiopathology ; Erectile Dysfunction ; etiology ; pathology ; physiopathology ; Male ; Models, Biological ; Myocytes, Smooth Muscle ; cytology ; physiology ; Penile Erection ; Penis ; cytology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo culture rat corpus cavernosum smooth muscle cells in vitro using a modified tissue culture method.

METHODSFifteen male rats were randomized into 3 equal groups, namely enzyme digestion group, tissue culture group, and modified tissue culture group. The penis of the rats was separated carefully and cut into small pieces, and seeded onto culture flasks and cultured in complete medium consisting of DMEM containing 20% fetal calf serum at 37 degrees C; in a humidified atmosphere with 5% carbon dioxide. The cells growth was observed under phase contrast microscope and the smooth muscle cell specific proteins alpha-SM-actin and desmin were identified immunohistochemically.

RESULTSThe alpha-SM-actin-positive cell rate was 96.3% in rat corpus cavernosum smooth muscle and 23.8% in the fibroblasts, and the corpus cavernosum smooth muscle contained 74.4% desmin-positive cells while the fibroblasts showed no desmin positivity. Significant difference was found in the positive cell rate for desmin among the 3 groups, with the highest positive cell rate occurred in modified tissue culture group.

CONCLUSIONDesmin may serve as a marker for identifying corpus cavernosum smooth muscle cells. The modified tissue culture method can result in highly purified corpus cavernosum smooth muscle cells with intact structure and functions.

Actins ; analysis ; Animals ; Biomarkers ; analysis ; Cell Proliferation ; Desmin ; analysis ; Male ; Muscle, Smooth ; cytology ; Penis ; cytology ; Rats ; Rats, Sprague-Dawley ; Tissue Culture Techniques

OBJECTIVETo explore the effect of 18-β glycyrrhetinic acid (GA) on the endoplasmic reticulum of nasal epithelial cells in allergic rhinitis (AR) model rats.

METHODSTotally 96 Wistar rats were randomly divided into the blank group, the AR model group, the loratadine group, the GA group, 24 in each group. AR models were established by peritoneally injecting ovalbumin (OVA). Morphological scoring was performed. GA at 21. 6 mg/kg was intragastrically administered to rats in the GA group. Nasal mucosal tissues were taken for electron microscopic examinations at the second, fourth, sixth, and tenth week after drug intervention.

RESULTSThe overlapping score was 2.10 ± 0.45 in the blank group, 5.10 ± 0.56 in the loratadine group, 5.10 ± 0.56 in the AR model group, 5.20 ± 0.78 in the GA group, showing statistical difference when compared with the blank group (P < 0.01). Results under transmission electron microscope showed that the number of the endoplasmic reticulum increased in the AR model group, with obvious cystic dilatation, a lot of vacuole formation, and degranulation. A large number of free ribosomes could be seen in cytoplasm. With persistent allergen exposure, changes mentioned above was progressively aggravated in the endoplasmic reticulum of nasal mucosal epithelium in the AR model group. But the dilation of endoplasmic reticulum, vacuole formation, and degranulation were relieved in the GA group, and got close to those of the blank group.

CONCLUSION18-β GA could improve the expansion, vacuolization, and degranulation of the endoplasmic reticulum of nasal epithelial cells in AR model rats.

Animals ; Anti-Inflammatory Agents ; pharmacology ; therapeutic use ; Endoplasmic Reticulum ; Epithelial Cells ; drug effects ; Glycyrrhetinic Acid ; pharmacology ; therapeutic use ; Nasal Mucosa ; drug effects ; Rats ; Rats, Wistar ; Rhinitis, Allergic ; drug therapy

OBJECTIVETo investigate the radiosensitizing effect of nitric oxide (NO) combined with radiation on esophageal cancer cell line TE-1.

METHODSMethyl thiazolyl tetrazolium (MTT) assay was used to assess the effects of NO and radiation on TE-1 cells regarding inhibition of cell proliferation. Flow cytometry was used to examine the effect of NO and radiation on cell apoptosis and cycle. Reverse transcription polymerase chine reaction and Western blot were used to evaluete the effect of NO on mRNA and protein expression of manganese superoxide dismutase (MnSOD).

RESULTSNO inhibited the proliferation of TE-1 cells while significantly enhancing their radiosensitivity. The application of NO combined with radiation significantly increased the apoptosis rate and G2/M phase proportion of TE-1 cells, with substantial decreases in the MnSOD mRNA and protein expression levels.

CONCLUSIONSNO reduces the MnSOD mRNA and protein expression levels by affecting TE-1 cell cycle, further inhibiting the apoptosis of esophageal cancer cells and enhancing the killing effect of radiation on esophageal cancer cells.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Esophageal Neoplasms ; drug therapy ; metabolism ; pathology ; Humans ; Nitric Oxide ; therapeutic use ; Radiation Tolerance ; drug effects ; Superoxide Dismutase ; metabolism