Adult ; Endoscopy ; Female ; Humans ; Maxillary Sinus ; Neurilemmoma ; surgery ; Otorhinolaryngologic Surgical Procedures ; methods ; Paranasal Sinus Neoplasms ; surgery ; Temporal Bone ; innervation

The marine biological source of mineral drugs recorded in Chinese Pharmacopoeia (2010 version) mainly including pearl, nacre, clam shell, common oyster shell, ark shell, cuttle bone, and sea-ear shell are widely used in clinical. Calcium carbonate and a small amount of protein are the main components in this type of drugs. In this paper, a systematical and comparable study were carried out by determination of calcium carbonate by EDTA titration method, the crystal of calcium carbonate by X-Ray powder diffraction and the total amino acids (TAAs) of the hydrolyzed samples by ultraviolet spectrophotometry method. As a result, the crystal structure is calcite for common oyster shell, mixture of calcite and aragonite for nacre and sea-ear shell, aragonite for the other drugs. The content of calcium carbonate ranged from 86% to 96%. Cuttle bone has the highest amount of TAAs among the seven drugs which reached 1.7% while clam shell has the lowest content of 0.16% on average. In conclusion, an effective method was developed for the quality control of marine mineral drugs by comprehensive analysis of calcium carbonate and TAAs in the seven marine mineral drugs.
Amino Acids ; analysis ; chemistry ; Animal Shells ; chemistry ; Animals ; Calcium Carbonate ; analysis ; chemistry ; Crystallization ; Edetic Acid ; chemistry ; Mollusca ; chemistry ; classification ; Pharmaceutical Preparations ; analysis ; chemistry ; standards ; Quality Control ; Reproducibility of Results ; Seawater ; Species Specificity ; Spectrophotometry, Ultraviolet ; X-Ray Diffraction

OBJECTIVETo investigate the effect of serum contained Chinese drugs for nourishing Shen and activating blood (S-NSAB) on activity of purified retinal ganglion cells (RGCs) cultured in high glucose medium.

METHODSPurified RGCs of SD rats were cultured in stimulative stable high glucose (50 mmol/L) condition (SHG) and fluctuated glucose condition (FGC) separately, they were intervened with S-NSAB, and the lactate dehydrogenase (LDH) leakage was detected by spectrophotometer for estimating the activity of RGCs.

RESULTSLDH leakage (U/L) in SHG culture was 1 349.17 +/- 215.50 at 24 h, 1220.24 +/- 124.53 at 48 h and 1982.14 +/- 219.03 at 72 h, all significantly lower than that in normal control at the corresponding time points (1628.10 +/-122.10, 1484.13 +/- 127.55 and 2155.75 +/- 140.44, respectively, P<0.05), whereas it was obviously higher in FGC culture at 72 h (2299.60 +/- 88.35), showing that LDH leakage in FGC was significantly higher than that in SHG at the corresponding time points (P<0.05). The LDH leakage was obviously decreased by Chinese medicine intervention with S-NSAB both in SHG at 72 h (1797.62 +/- 146.40) and in FGC at 48 h (1259.92 +/- 87.74) and 72 h (1940.40 +/- 155.47), the difference between pre- and post-intervention was significant (P<0.05).

CONCLUSIONFluctuated glucose conditions of culture medium could obviously damage the membranous stability of RGCs to enhance their permeability and lower the activity of cells; S-NSAB could improve these abnormalities in either SHG or FGC condition, which may be one of the important mechanisms of Chinese formula for nourishing Shen and activating blood in preventing and treating diabetic retinopathy.

Animals ; Animals, Newborn ; Cell Survival ; drug effects ; Cells, Cultured ; Culture Media ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Retinal Ganglion Cells ; cytology ; drug effects ; Serum

BACKGROUND:The intracel ular accumulation of reactive oxygen species leads to oxidative stress. Hypoxia is widespread in physiological and pathological condition. Variation of bone proliferation and differentiation when bone tissues cultured or bone cel s induced toxicity by reactive oxygen species under hypoxia have not yet been reported. OBJECTIVE:To observe the biological characteristics of MC3T3-E1 pretreated with different concentrations of hydrogen peroxide (H2O2) in hypoxia, thus understanding the cel mechanism underlying prolonged bone healing in the elderly with osteoporosis and diabetes. METHODS:The MC3T3-E1 cel s pretreated with different concentrations of H2O2 were cultured in different oxygen concentrations. The proliferation of MC3T3-E1 was detected by cel counting kit-8. The cel differentiation was detected through alkaline phosphatase staining and alizarin red staining. Total RNAs were extracted and used for analyzing the mRNA levels of col age type 1, alkaline phosphatase and Cbfa1. RESULTS AND CONCLUSION:When MC3T3-E1 pretreated with 200μmol/L H2O2 for 6 hours, the cel proliferation was increased with time, but lower than that in the control group. The alkaline phosphatase activity was weakened, and the number of mineralized nodes was decreased at the early stage of differentiation. When MC3T3-E1 pretreated with 400μmol/L H2O2 for 6 hours, the cel proliferation was decreased obviously. The alkaline phosphatase activity was stil weakened, and the number of mineralized nodes was decreased further, but not affected by hypoxia. When MC3T3-E1 pretreated with 400μmol/L H2O2 for 6 hours and then cultured in hypoxia, the mRNA expression of Cbfa1 was decreased, but the mRNA expressions of col age type 1 and alkaline phosphatase were significantly increased. These results suggest that MC3T3-E1 pretreated with low concentration of H2O2 show a significant decrease in proliferation, while MC3T3-E1 pretreated with a high concentration of H2O2 and cultured in hypoxia show a decrease in osteogenic differentiation, especial y at the early stage of alkaline phosphatase formation.

To investigate the biological role of snake venom cystatin(sv-cystatin) in tumor progression, the cDNA of sv-cystatin amplified by PCR from pUC18-cystatin plasmid was cloned into methanol-inducible expression vector pPICZ?A. The linearized recombinant plasmid pPICZ?A-cystatin was transfered into Pichia pastoris, strain GS115 by electrophoration. Transfermants with phenotype Mut+ selected were identified by PCR analysis and induced in 1.0% methanol. The reombinant sv-cystatin protein was examined by SDS-PAGE, Western blot analysis. The molecular mass of expression product was about 14 kDa and approximately 16 mg/L of recombinant sv-cystatin was produced from one of GS115-cystatin transformants. The chromatography purified protein could reduce the activity of papain. The ability of B16F1 cells treated with recombinant sv-cystatin to invade the reconstituted basement membrane decreased significantly (P

Adult ; Ethmoid Sinus ; Female ; Humans ; Neurofibroma ; Paranasal Sinus Neoplasms

OBJECTIVETo investigate the impacts of steady high-glucose or fluctuated glucose conditions on glutamate (Glu) release in purified retinal ganglion cells (RGCs) cultured in vitro, and the effect of serum contained Chinese drugs for nourishing Shen and activating blood (S-NSAB) on it.

METHODSRGCs of neonatal SD rats were cultured by antibody combined two-step purified method in different conditions: the simulated normal condition, the steady high-glucose condition and the fluctuated glucose condition, and they were intervened with S-NSAB. Thereby, the experiment was carried out in 6 groups, i.e. the normal control group (A), the S-NSAB intervened group (B), the steady high-glucose cultured group (C), the steady high-glucose cultured and S-NSAB intervened group (D), the fluctuated glucose cultured group (E), and the fluctuated glucose cultured and S-NSAB intervened group (F). Content of Glu in the extracellular fluid was detected at 24, 48 and 72 h after intervention with a full-automatic biochemical analyzer. And the data obtained were statistically analyzed with SPSS 13.0 soft ware.

RESULTSRelease of Glu at 24 h after intervention in Group E (256.33 +/- 25.73 mg/L) was obviously higher than that in Group A and Group C (134.22 +/- 9.14 mg/L and 141. 17 +/- 22.13 mg/L, P < 0.05); at 24 h and 72 h in Group B (124.50 +/- 10.30 mg/L and 30. 17 +/- 2.97 mg/L) was obviously lower than in Group A respectively (P < 0.05); in Group D at 24 h (127.50 +/- 16.94 mg/L), 48 h (26.17 +/- 3.99 mg/L) and 72 h (27.67 +/- 3.49 mg/L) were lower than in Group C; in Group F at 24 h (228.33 +/- 18.41 mg/L) and 72 h (28.00 +/- 2.41 mg/L) were lower than in Group E respectively at the corresponding time points.

CONCLUSIONSFluctuated glucose condition could obviously increase the Glu release of RGCs, to cause extracellular large amount Glu accumulation, which induces the exciting neurotoxicity to RGCs and finally to aggravate the injury on cells. S-NSAB could reduce the Glu release to some extent in the steady-high or fluctuated glucose conditions, diminish the injury of RGCs from exciting neurotoxicity of Glu, and it might be one of the intervening pathways of Chinese drugs for NSAB in preventing and treating DRP.

Animals ; Animals, Newborn ; Cells, Cultured ; Diabetic Retinopathy ; prevention & control ; Drugs, Chinese Herbal ; pharmacology ; Glucose ; pharmacology ; Glutamic Acid ; metabolism ; Rats ; Rats, Sprague-Dawley ; Retinal Ganglion Cells ; cytology ; metabolism

Objective To compare the different effects of 1% diatrizoate meglumine,2.5% mannitol and water as oral contrasts in PET/CT scan in gastrointestinal tract delineation and 18F-fluorodeoxyglucose (FDG) uptake. Methods Sixty-one patients referred for PET/CT scan without gastrointestinal diseases were divided into three groups randomly ( random number method). One liter of 1% diatrizoate meglumine,2.5% mannitol,or water was orally taken by groups 1 (25 cases),2 (20 cases) and 3 ( 16 cases),respectively before scan. The scan was performed with GE Discovery LS PET/CT scanner in two-dimensional (2D) mode 50 min after 18F-FDG (5.55 MBq/kg) injection. Patients with abdominal lesions were excluded from this study. The degree of gastrointestinal filling and 18F-FDG uptake was evaluated by 3 nuclear medicine physicians using visual analysis according to a 4-grade classification method:none,mild,moderate,and high. Statistically analysis was performed by Kruskal-Wallis,Mann-Whitney and paired t tests.Results Both the differences of serum glucose and insulin levels were not significant before and after contrast taken in group 2. Group 2 had better gastrointestinal filling than that of group 1 and also better than group 3 except in rectum. The stomach,jejunum,ascending,and transverse colon were better filled in group 1 than in group 3. The degree of 18F-FDG uptake of group 3 was significantly higher than that of group 2 in stomach,jejunum and ileum (z= -3. 192,-3.290,-3.290,all P＜0.05),and was also significantly higher than that of group 1 (z = - 3. 603,P ＜ 0.05) in jejunum. The degree of 18 F-FDG uptake of group 3 was significantly lower than that of group 1 in ascending colon (z = - 2. 706,P ＜ 0. 05 ) and was significantly lower than that of group 1 and 2 in transverse and descending colon (z= - 3. 503,- 2.403,- 4.225,-4. 027,all P ＜0.05),and was also significantly lower than that of group 2 in rectum (z = -4. 128,P ＜0. 01 ). The maximum CT values in stomach,jejunum,ileum and ascending colon in group 1 were ( 132 ±23),(191 ±31),(313 ±47) and (374±53) HU,respectively,whose difference was significant (t = -7.088--1.781,all P ＜0. 01 ). Conclusion Oral iso-osmotic mannitol intake has better gastrointestinal filling and less physiological 18F-FDG uptake compared to diatrizoate meglumine and water.

Adult ; Aged ; Aged, 80 and over ; Colles' Fracture ; therapy ; Combined Modality Therapy ; External Fixators ; Female ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged

OBJECTIVETo investigate the effect of suppression of CCL5 ligand gene on the proliferation of human breast cancer cells.

METHODSA lentiviral vector carrying a short interfering RNA (siRNA) targeting CCL5 was transfected into human breast cancer cell line MCF-7 and MDA-MB-231 cells. The expression of CCL5 mRNA in the cells was detected by real-time PCR. The proliferation of MCF-7 and MDA-MB-231 cells was assessed by MTT assay and FACS assay, and the colony formation ability of both cell lines were measured, respectively.

RESULTSReal time PCR showed a good knockdown effect of CCL5 in both cell-lines. Colony-forming assay showed that the ability of colony formation of MCF-7/CCL5-siRNA and MDA-MB-231/CCL5-siRNA was decreased markedly. The colony number of MCF-7/CCL5-siRNA group was (0.34 ± 0.08), significantly lower than 0.81 ± 0.12 in the MCF-7/CCL5-N group and 0.92 ± 0.12 in the MCF-7 group (P < 0.05). The colony number of MDA-MB-231/CCL5-siRNA group was 0.33 ± 0.10, significantly lower than 0.97 ± 0.09 in the MDA-MB-231/CCL5-N group and 1.04 ± 0.07 in the MDA-MB-231 group (P < 0.05). However, MTT assay revealed that the proliferation of MCF-7/CCL5-siRNA cells was not significantly different from that of MCF-7/CCL5-N or MCF-7 cells, respectively (P > 0.05), and the same result was found in MDA-MB-231 cells. FACS assay showed that the proliferation index (PI) of groups MCF-7/CCL5-siRNA, MCF-7/CCL5-N and MCF-7 were 0.48 ± 0.03, 0.43 ± 0.01 and 0.45 ± 0.02. The PI of groups MDA-MB-231/CCL5-siRNA, MDA-MB-231/CCL5-N and MDA-MB-231 cells were 0.48 ± 0.02, 0.44 ± 0.05 and 0.47 ± 0.02. There was no statistical difference among them (P > 0.05).

CONCLUSIONThe down-regulation of CCL5 gene in human breast cancer cells may significantly suppress their colony formation ability, rather than affecting their population doubling time to some extent.

Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Chemokine CCL5 ; genetics ; metabolism ; physiology ; Down-Regulation ; Female ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection