Objective To evaluate the protective effect of sinomenine (SIN) on renal ischemia/reperfusion (I/R) in mice. Methods In the experiment one, 12 C57BL/6 mice were randomly divided into 2 groups: SIN group (mice were injected with 200 mg/kg SIN by tail vein) and control group (mice were injected with equal volume of saline). Six and 24 hs later, the serum was collected and the contents of alanine aminotransferase (ALT) and creatinine (SCr) were determined. In the experiment two, C57BL/6 mice were randomly divided into 3 groups: sham-operated (SO) group, SIN group (mice were injected with 200 mg/kg sinomenine just before ischemia induction) and saline group (mice were injected with equal volume of saline at the same time). At the 6th h after reperfusion, the sera and renal samples subject to IR injury were collected. The SCr and BUN levels in serum were determined and renal histological changes were also examined. The apoptosis of renal tubular epithelial cells was measured by using terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assay. The infiltration of F4/80 positive macrophages was measured by using immunohistochemistry and that of neutrophils with myeloperoxidase (MPO) kits. The mRNA expression of tumor necrosis factor (TNF)-α, chemokine CXC ligand (CXCL)-10, intercellular adhesion molecule (ICAM)-1 and IL-17 was detected by using real-time reverse transcription PCR. The activation of transcription factor NF-κB was measured by using Western blotting. Results In the experiment one, there was no significant difference in ALT and SCr between the two groups at 6 or 24 h. In the experiment two,levels of SCr and BUN were lower in SIN group (P＜0. 05 or P＜0. 01 ), histological damage was milder (P＜0. 01 ), and apoptosis rate of renal tubular epithelial cells apoptosis was lower than in saline group (P＜0. 05). The infiltration of macrophages, neutrophils and the mRNA expression of TNF-α, CXCL-10, ICAM-1 and IL-17 in the renal tissue in SIN group were reduced as compared with saline group (P＜0. 05 or P＜0. 01 ). The activation of NF-κB in SIN group was significantly downregulated as compared with saline group. Conclusion SIN can ameliorate the renal IR injury without hepatic or renal toxicity, which is associated with inhibition of acute inflammatory response induced by reperfusion.

Objective To investigate the mechanism of Bushentongluoshugan Formula's effect in penile erection.Method All 40 adult male rats were randomly divided into the control group, castrated group, Bushentongluoshugan Formula group and sterandryl group.The rats of all groups, except the control group, were ablated bilateral testes. Bushentongluoshugan Formula group and sterandryl group were given the formula and sterandryl respectively. After 3 weeks the penile tissue was collected and the expression of transforming growth factor-β-mRNA (TGF-β-mRNA) was detected by using polymerase chain reaction (PCR) technique.Result The expression of TGF-β-mRNA in the castrated group was higher than that in other three groups, and among them the incremental order was the control group, sterandryl group and Bushentongluoshugan Formula group according to the decreasing performance of TGF-β-mRNA expression. The internal control of TGF-β-mRNA was less in the control group, sterandryl group and Bushentongluoshugan Formula group with a significant difference compared with that in the castrated group (P<0.01), and there was no significant difference between the sterandryl group and Bushentongluoshugan Formula group (P>0.05).Conclusion Bushentongluoshugan Formula plays an important role in penile erection through regulating the expression of TGF-β-mRNA.

OBJECTIVEConditioned taste preference (CTP) is a taste learning reflex by which an animal learns to prefer a substance which tastes not well and has been studied with much interest in recent years. However, the neural substrates of CTP are less known. This study aimed to determine the possible neural path- ways of CTP and whether serum leptin level and the leptin receptor (OB-Rb) in the hind brain are involved following CTP formation.

METHODSWe established CTP of quinine in rats with a 2-bottle preference test. The serum leptin concentrations were detected, the expression of c-fos in the rat brain was tested to determine the nuclei in relation with establishment of CTR Finally, the OB-Rb mRNA expression was examined by RT-qPCR assay in parabrachial nucleus (PBN) and the nucleus of the solitary tract (NST) of the hind brain.

RESULTSCompared with control group, the level of serum leptin was higher in the CTP group (4.58 ± 0.52 vs 1.67 ± 0.25 µg/L, P < 0.01); increased c-fos positive cells were found in the anterior hypothalamus (AH, 221.75 ± 4.96 vs. 178.50 ± 6.63 cells/mm², P < 0.05), the basal lateral amygdala (BLA, 70.75 ± 6.17 vs 56.50 ± 3.62 cells/ mm², P < 0.05) and the nucleus of the solitary tract (NST, 41.25 ± 1.32 vs 32.50 ± 1.02 cells/mm², P < 0.05). But in ventromedial nucleus of the hypothalamus (VMH, 20.75 ± 2.73 vs 38.5 ± 1.54 per 1 mm², P < 005), PBN (21.50 ± 2.24 vs 36.25 ± 1.49 cells/mm², P < 0.05) and the central nucleus of the amygdala (CeA, 22.25 ± 1.53 vs 35.50 ± 2.11 cells/mm², P < 0.05), the number of c-fos positive cells was decreased in the CTP group. In addition, we found OB-Rb mRNA expression in PBN of CTP group rats was higher than that of control group (0.95 ± 0.055 vs 0.57 ± 0.034, P < 0.05), while there was no significant difference of OB-Rb mRNA expression in NST between the two groups.

CONCLUSIONNuclei AH, BLA, NST, VMH, PBN and CeA participate in the formation of CTP. Leptin and its receptor in PBN may be involved in the formation and maintenance of CTP.

Animals ; Conditioning (Psychology) ; Leptin ; blood ; Rats ; Receptors, Leptin ; physiology ; Rhombencephalon ; physiology ; Taste ; physiology

BackgroundHuman retinal gliocytes play an important role in proliferative diseases,which are the basis of in vitro studies.Researchers have cultured human retinal gliocytes in the past.In our study,we found that the cells we cultured presented a unique shape different from those by other researchers.ObjectiveThis study was to design to produce a new culture and purification method for retinal gliocyte in vitro.Methods Retinal tissue was isolated from human eyeballs and digested using the two-step digestion method (2％ pancreatin and 0.133％collagenase Ⅵ) to harvest the retinal glio cytes.The cells were collected and cultured in endothelial cell-targeted nutrient culture containing 10％ fetal calf serum and supplemented with β-endothelial cell growth factor (ECGF) and sodium heparin,and the culture dishes were coated with fibronectin(FN) to promote the attachment of retinal gliocyte.During the culturing process,the gliocytes were identified by the observation of morphological characteristic and regular histological examination.The identification of the cells also was performed by immunochemistry targeting glial fibrillary acidic protein (GFAP),Vimentin,neuron specific enolase ( NSE ),S-100,CD34,and Ⅷ factor.Results Retinal gliocytes were isolated successfully from the human retina by the two-step digestion method.Primary cultured cells attached after 72 hours and achieved confluency between day 9 and 10 that were aligned petaliform in shape.Regular histological examination after H&E staining showed blue cell nuclei and light red cytoplasm.The target cells presented with strong responses for GFAP and Vimentin and no response for NSE,S-100,CD34 and Ⅷ factor.ConclusionsLarge amount of purified human retinal gliocytes can be obtained by two-step digestion and cultured in endothelial cells-targeted culture medium supplemented with β-ECGF and sodium heparin in plates coated with FN.The cultured cells expressed markers for retinal gliocytes.However,specific features of these cells remain to be further elucidated.

OBJECTIVETo explore the clinical features and diagnosis of obstructive sleep apnea syndrome (OSAS) in children.

METHODSSixty children with OSAS were reviewed, every patient was monitored with polysomnography (PSG) for 7 hours at night for 11 parameters, including the longest apnea time (LAT), apnea and hypopnea index (AHI), the lowest oxygen saturation (SaO(2)), and snore index etc., the parameters of the 2 groups were compared. Meanwhile, tonsillectomy and adenoidectomy were performed for 40 cases of OSAS, and the parameters obtained before and after operation were analyzed.

RESULTSAdenotonsillar hypertrophy and the loud snoring during sleep were found in all cases. The mean values of the PSG parameters were as follows: the longest apnea time was 53 (8-178) seconds (s); the total time of apnea was 310.5 (26-5,260) s; the time of apnea was 26 (3-240) s; the longest hypopnea time was 41 (5-94) s; the total times of hypopnea was 170 (5-2,860) s; the time of hypopnea was 10 (1-85); the apnea index was 4.1 (0.5-25.9); the hypopnea index was 1.4 (0-16.1); the apnea and hypopnea index (AHI) was 6.8 (0.5-38.2); the snore index was 81.7 (1.3-414.8); the lowest saturation of oxygen was 0.78 (0.25-0.93). There was not statistically significant difference in the parameters between 2-7 year group and over 7 year group (P > 0.05). The parameters of postoperation group: the mean value of the longest apnea time was 15.5 (0-60) s; the total time of apnea was 56.4 (60-205) s; the time of apnea was 10.33 (0-40); the longest hypopnea time was 13.25 (0-30) s; the total times of hypopnea was 44.6 (0-73); the hypopnea time was 4.32 (0 - 30) s; the apnea index was 0.6 (0-12); the hypopnea index was 0.62 (0-4); the apnea and hypopnea index (AHI) was 1.25 (0.1-12); the snore index was 30.08 (1.8-102); the lowest oxygen saturation was 93.5% (64%-97%). Compared with preoperation groups there was a statistically significant difference (P < 0.01). Clinically effective rate of the surgeries was over 90%.

CONCLUSIONAdenotonsillar hypertrophy seemed to be an important cause of OSAS in children. Snoring, dyspnea, apnea and low ventilation are the major clinical characteristics of OSAS in children. Confirmed diagnosis of the syndrome in children requires PSG recordings.

Child ; Child, Preschool ; Female ; Humans ; Male ; Monitoring, Ambulatory ; Polysomnography ; Sleep Apnea, Obstructive ; diagnosis ; physiopathology ; Time Factors

Adult ; Antiviral Agents ; administration & dosage ; adverse effects ; Drug Therapy, Combination ; Female ; Follow-Up Studies ; Hepatitis C, Chronic ; drug therapy ; genetics ; Humans ; Interferon Type I ; administration & dosage ; adverse effects ; Interferon-alpha ; Male ; Middle Aged ; RNA, Viral ; blood ; Recombinant Proteins ; Ribavirin ; administration & dosage ; adverse effects ; Safety ; Treatment Outcome ; Viral Load

OBJECTIVETo prepare the superparamagnetic iron oxide (SPIO)-labeled antisense oligodeoxynucleotide (ASODN) probe and evaluate the application of this probe in cellular magnetic resonance imaging (MRI).

METHODSWe prepared the SPIO-labeled ASODN probe using chemical cross linking method to conjugate SPIO to ASODN, detected its configuration by atomic force microscopy, determined the conjugating rate and biology activation by high performance liquid chromatography, and detected the stability by polyacrylamide gel electrophoresis. After that, we transfected the SK-Br3 oncocytes which had over-expression of the c-erbB2 oncogene by this probes, observed the intracellular iron distribution by optical microscope, measured iron content by atomic absorption spectroscopy, and observed the signal change by MRI.

RESULTSAtomic force microscope showed that the SPIO-labeled ASODN probe was mostly spherical and well-distributed, with a diameter of 25-40 nm and a conjugating rate of 100%. This probe had inhered biological activity and stability. In addition, light microscopy revealed an intracellular uptake of iron oxides in the transfected SK-Br3 oncocyte, and the iron content of the group of transfected SK-Br3 oncocytes was significantly higher than those of other contrast groups (all P < 0.01). MRI showed that transfected SK-Br3 oncocyte had the lowest signal among all other cells (all P < 0.05).

CONCLUSIONSWe prepared the SPIO-labeled ASODN probe successfully. It can effectively transfect SK-Br3 oncocyte and enter SK-Br3 oncocyte, and thus reduce the signal intension in MRI.

Cell Line, Tumor ; DNA, Antisense ; chemistry ; genetics ; Ferric Compounds ; chemistry ; Humans ; Magnetic Resonance Imaging ; Magnetics ; Molecular Probe Techniques ; Oligodeoxyribonucleotides ; chemistry ; genetics ; Oxyphil Cells ; chemistry ; Receptor, ErbB-2 ; analysis ; genetics

Adult ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; diagnostic imaging ; physiopathology ; surgery ; Humans ; Male ; Middle Aged ; Recovery of Function ; Scapula ; diagnostic imaging ; injuries ; physiopathology ; surgery ; Tomography, X-Ray Computed

Background The incidence of retinal vascular diseases increase annually,such as diabetic retinopathy,retinopathy of prematurity and age-related macular degeneration.The key of treatment for these diseases is how to evaluate retinal vascular change effectively and objectively.Retro-orbital injection of fluorescein isothiocyanatedextran (FITC-dextran) is a simple and effective method for observing C57BL/6J mouse retinal vessels.But,whether it is suitable for other mice and rats is seldom reported.Objective This experiment was to assess the feasibility of the observation of retinal vessels by retro-orbital injection of FITC-dextran in different genus of mouse and offer the reference for relevant study.Methods Twelve animals of C57BL/6J mice,Kunming mice,SD rats and Wistar rats were selected,respectively and divided into the experimental group and control group at average.The right eyes of the animals of the experimental group received the retro-orbital injection of 9 ml/kg FITC-dextran,and the right eyes of animals of the control group received PBS solution at the same volume and way.All the animals were sacrificed 10 seconds after injection and both eyes of each animal were obtained for retinal stretched preparation.The retrobulbar tissue and whole-mount retina were viewed under a fluorescence microscope.The use of the animals complied with Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results Retinal blood vessels labeled by FITC-dextran could be observed in both eyes of C57BL/6J mice and Kunming mice to present with a green fluorescence in experimental group under a fluorescence microscope,but no any fluorescence-labeled retinal blood vessel was exhibited in the control mice.The retinal blood vessel could not be observed in all eyes of SD rats and Wistar rats after the injection of FITC-dextran both in the experimental group and the control group under a fluorescence microscope.The surrounding tissues of the right eyes of mice and rats dyed with green fluorescence of FITC-dextran in the experimental group,however,green fluorescence could not be seen in the surrounding tissues of the left eyes of mice and rats.Conclusions Retro-orbital injection of FITC-dextran is a suitable method of observing the retinal vessels of mouse but not rat.

OBJECTIVETo assess the clinical effects of (125)I interstitial brachytherapy for malignant tumors.

METHODSOne hundred and twelve patients with malignant tumors of stage II stage and III under went radical resection combined with (125)I intraoperative implantation. Seven days and three month after operation WBC count and immune markers were observed. Blood biochemistry ultrasonography and X-ray were performed to observe recurrence and metastasis of tumors per three months.

RESULTSIn the 112 patients, 3 died of tumor recurrence in six months, and others survived with the longest time of 35 months.

CONCLUSION(125)I interstitial brachytherapy for malignant tumor is simple, safe and effective.

Adult ; Aged ; Brachytherapy ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Humans ; Iodine Radioisotopes ; therapeutic use ; Male ; Middle Aged ; Neoplasms ; radiotherapy ; therapy ; Retrospective Studies ; Treatment Outcome