OBJECTIVE: To explore the expression levels of three T-type calcium channel receptors (α1G; α1H; α1I) in the cochlea and spiral ganglion neurons of C57BL/6J mice with different ages. METHOD: Thirty cases of C57BL/6J mice were divided into three groups (6-8 W, 24-26 W, 42-44 W) according to the age. The expressions of three T-type calcium channel receptors were quantified by RT-PCR after hearing thresholds measured by ABR. RESULT: Three receptors were detected in the cochlea and spiral ganglion neurons of 6-8 W C57BL/6J mice. The quantitative results showed that the expression levels of α1H and α1I were highest among three receptors in spiral ganglion neurons and in the cochlea respectively. The expression levels of three receptors significantly decreased with age,especially at the age of 4244 W. CONCLUSION The expression of T-type calcium channel receptors reduced with age in the inner ear of C57BL/6J mice.
Animals ; Calcium Channels ; Calcium Channels, T-Type ; biosynthesis ; Cochlea ; metabolism ; Ear, Inner ; Mice ; Mice, Inbred C57BL ; Neurons ; Receptors, Calcium-Sensing ; Spiral Ganglion ; metabolism

AIM: To develop a method to detemine pinostrobin in Weitengning Talets (Lindera reflexae Hemsl.) by HPLC. METHODS:A C_(18) column was used,methyl alcohol-water(80∶20) was used as a mobile phase and the wavelength of UV detector was set at 290 nm. RESULTS:The linearity of this method was good with the average recovery of 97.9%,RSD was 0.74%(n=5). CONCLUSION:The methed is simple,reliable,sensitivity,and with good reproducibility.It can be used in quality control of Weitengning Tablets.

This research is to establish an HPLC method for the simultaneous determination of ophiopogonin D, ophiopogonin D', ophiopogonin C, deacetylophiopojaponin A and ophiogenin-3-O-α-L-rhamnosyl-(1-->2)-β-D-glucoside in Ophiopogonis Radix. HPLC-ELSD analysis was performed on a Kromasil 100-5 C₁₈ column (4.6 mm x 250 mm, 5 µm), with the mobile phase of acetonitrile (A) -water (B) in gradient elution mode (0-45 min, 35%-55% A), at a flow rate of 1 mL · min⁻¹. The column temperature was 35 °C and the drift tube temperature was 100 °C in a gas flow rate of 3.0 L · min⁻¹. The result showed that baseline of all the 5 constituents was well separated, and every constituent had wide linearity range and good linear relation (r > 0.999). The recovery rate was between 95.75% and 103.1%. The new established method for simultaneous determination of saponin constituents in Ophiopogonis Radix was sensitive and has good, repeatability. It could be applied to quality evaluation of Ophiopogonis Radix.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ophiopogon ; chemistry ; Plant Roots ; chemistry ; Saponins ; chemistry ; isolation & purification

Objective To explore the neurobehavioral change of mice from dams with human cytomegalovirus(HCMV) infection during first trimester. Methods Eight-week-old fertilized female Kunming mice were randomly divided into infected group and control group.On the 4th gestation day mice in infected group were injected intraperitoneally with 0.5 mL HCMV (1?10-6 50 percent of tissue cultured infective dose),and those in control group were injected intraperitoneally with 0.5 mL supernatant of cultured human fibroblast.Caesarean birth operation was performed on 3 randomly chosen fertilized mice before delivery. Fetuses were observed and their brain tissue were collected and analyzed under light and electron microscope separately.PCR test was used to determine HCMV pp65 antigen of offspring′s sera.Neurobeha-vioral test such as Morris Water Maze and Lashley Ⅲ Water Maze were performed on offspring mice of 6-7 weeks old.Results Compared with control group,the pathological changes such as degeneration,necrosis,and nucleus disappearance of nerve cells and giant cells were found in offspring′s brain of mice in infected group under light microscope. Under electron microscope,swelling of nerve cells and spherical virus particle in the cytoplasm were found in the brain of mice in infected group. HCMV pp65 antigen was detected in 7 offspring mouse′s se-rum in infected group.Offspring′s swimming time and speed were(30.21?12.74) s and(19.10?1.90) cm/s in infected group,while those in control group were (11.87?3.62) s and (23.21?1.02) cm/s by Morris Water Maze test,there were significantly differences between 2 groups (Pa

Background Ostrich acellular corneal stroma possesses a similar constitution to human corneal stroma,so it is expected to become one of ideal biological corneal carriers.Objective This study was to investigate the immunogenicity of acellular stroma carrier of ostrich cornea and offer the information for the development of industrialization and clinical use of acellular stroma carrier of ostrich cornea.Methods Twenty fresh ostrich eyeballs and 20 porcine eyeballs were collected.Acellular corneal stroma carriers of ostriches and pigs were prepared using low temperature freezing joint enzyme digestion method and desiccant dehydration method and sterilized by cobalt-60 irradiation.The corneal stroma carriers were preserved using drying and dehydration method.Forty-five male BALB/c mice were randomly divided into the sham operation group,ostrich acellular corneal stroma group and porcine acellular corneal stroma group.Acellular corneal stroma carriers of ostriches and pigs(wet weight after rewatering was 10 mg/piece) were subcutaneously implanted to the back of BALB/c mice,respectively.Wound healing and inflammatory response on the operative site were observed,and phenotype and activating rate of CD4+,CD8+ and CD25+in peripheral blood of mice were dynamically detected 7,14 and 28 days after ectopically implantation of heterogeneous corneal stroma by immunofluorescence labeling and flow cytometry analysis.Results No swelling and exudation were seen in the skin of operative site of the mice with a good healing of wound after surgery.There were no significant differences in the activating rates of CD4+,CD8+ and CD25+ cells in the peripheral blood of mice among the sham operation group,the porcine acellular corneal stroma group and ostrich acellular corneal stroma group in the three time points after surgery(CD4+:F=0.74,P=0.50;F=0.39,P=0.05;F=3.46,P=0.58.CD8+:F=1.75,P=0.21 ;F=1.14,P=0.35;F=0.78,P=0.48.CD25+:F=0.52,P=0.61 ;F=3.53,P=0.62;F=2.42,P=0.13).Conclusions The ostrich acellular heterogeneous corneal stroma carrier possesses low immunogenicity.It is inferred that ostrich acellular corneal stroma carrier can be used in heterogeneous corneal transplantation.

Background Fungal corneal ulcer is a visual-threatening eye disease,and drug therapy has a limiting efficacy.Corneal transplantation or eye enucleation sometimes is necessary to the severe patients.Corneal collagen cross-linking (CXL) is an effective method for some corneal diseases,but the study on CXL for fungal corneal ulcer is lack.Objective This study was to evaluate the clinical effectiveness and safety CXL for fungal corneal ulcer.Methods Fifteen 8-week-old healthy New Zealand white rabbits were used in this study and other 5 rabbits served as normal controls.Fungal corneal ulcer models were established in the right eyes of other 10 rabbits by infecting sickle bacteria liquid after corneal scratching and removing corneal epithelium,then decellularized ostrich corneal patch covered the defected cornea.The models were randomly divided into the non-treatment group and the CXL treatment group.Corneal lesions were examined under the slit lamp microscope every day,and cornea was pictured by laser scanning confocal microscope on the 3rd,7th,14th,21st and 28th day individually after CXL.All rabbits were sacrificed and corneal tissues were obtained 4 weeks after treatment,and the collagen fiber diameter and fibrocytes were observed under the scanning electron microscope.Results Fungal corneal ulcer models were successfully established by corneal scratching and decellularized ostrich cornea covering.The gray ulcer lesions and hypbae like bean pod were seen by slit lamp microscope and laser scanning confocal microscope 3 days after modeling.Corneal ulcer deepened and expanded 1 week later,and there were a large number of spore and hyphae criss-crossing as short rod in shallow stroma.Inflammatory cells were observed in corneal endothelial cells and ocular anterior chamber.In the CXL treatment group,the range of corneal epithelial deficiency was less than that in the nontreatment group on the 3rd,7th,14th,and 21st (all at P＜ 0.05).The diameters of collagen fibers were (24.6± 1.8) nm,(24.9 ± 1.9) nm and (43.0 ± 7.4) nm in the normal control group,non-treatment group and CXL treatment group,showing a significant difference among the 3 groups (F =27.05,P =0.00),and the collagen diameters were thicker in the CXL treatment group than those in the normal control group and non-treatment group (t =5.40,-5.30,both at P＜0.05),and fibrocytes were seen among the collagen fibers.No significant difference was found in the collagen diameters between the non-treatment group and normal control group,and the fibrocytes were less in the non-treatment group.Conclusions CXL therapy can treat fungal corneal ulcer by enhancing collagen,promoting fibrocytes proliferation,suppressing fungus and inflammatory response and accelerating tissue repair.

AlM:To study the refractive status characteristics aftser cataract surgery and the correlation between preoperative anterior chamber depth ( ACD) and refractive status.METHODS: Ninety-six cases of patients with cataract were randomly divided into two groups. The patients in phacoemulsification group were treated with phacoemulsification combined intraocular lens ( lOL ) implantation while the patients in small incision group were treated by small incision extracapsular cataract extraction combined with lOL implantation. Changes in ACD and postoperative refractive status and refractive fully corrected value were counted and the correlation of them were analyzed .RESULTS: ACD of the phacoemulsification group s deepened 0. 74mm while that of the small incision group deepened 0. 78mm after treatment and there was no significant difference (P>0. 05). After operation, the ACD of two groups significantly deepened ( P<0. 05 ). The postoperative visual acuity of two groups were significantly better than the uncorrected visual acuity of two groups (P<0. 05). The postoperative refraction of two groups patients were mainly 0 ~ +1. 0D ( 41. 67% and 54. 16%) and+1. 25~+2. 0D (43. 75% and 33. 33%) (P>0. 05). CONCLUSlON: ACD is significant deepened after operation. Surgeon needs full consideration of changes to improve the refractive lOL calculation accuracy.

Near-infrared (NIR) was used as rapid analysis method to identify the sulfur fumigated Puerariae Lobatae Radix. NIR spectra of the cross-sectional and longitudinal selection of samples were acquired. Principal component analysis was conducted. The samples were randomly selected. The different pretreatment methods were compared. Discriminant models were established for every type of spectra to calculate the recognition rate. The orthogonal test and nonparametric test were used to test data normality. The result showed that absorbance values of different sections were different due to the different structure, and the raw spectra were analyzed by PCA method. The result founded that the cumulative contribution rate was arrived at 99.2% while the PC numbers were arrived at 3. The pretreatment method based on the MSC + 1D + Savitzky-Golay was the best to establish the model. For the 50 models constructed with cross-section and longitudinal spectra and total spectra, the recognition rate were (94.4 ± 0.66)%, (94.4 ± 0.66)%, (95.3 ± 0.65)%, respectively, and no difference was observed. The NIR method could be used to identify the sulfur fumigated Puerariae Lobatae Radix.
Chemistry, Pharmaceutical ; methods ; Discriminant Analysis ; Drugs, Chinese Herbal ; chemistry ; Fumigation ; Pueraria ; chemistry ; Spectroscopy, Near-Infrared ; Sulfur ; chemistry

BACKGROUND: Rheumatoid factor (RF) is a kind of autoantibody which is routinely used as a factor in patients with rheumatoid arthritis (RA) to evaluate disease activity and severity.But in clinical practice, it occurs frequently that RF values do not decrease according to clinical improvement in RA patients. OBJECTIVE: To investigate the association between rheumatoid factor (RF) and activity or disease severity of RA.DESIGN, TIME AND SETTING: A randomized cross-sectional study was performed in the Department of Rheumatology and Immunology, the Third Affiliated Hospital of Sun Yat-sen University during September 2006 and September 2007.PARTICIPANTS: Seventy-six patients,65 females and 11 males,mean age of (44±13) years,with RA diagnosed according to the American College of Rheumatology (ACR) criteria for RA were included in this study. METHODS: Seventy-six patients with active RA were randomly recruited and assessed for functional status,radiographic change,joint pain,morning stiffness, tender joint count (TJC), tender joint score (TJS), swollen joint count (SJC), swollen joint score (SJS), Health Assessment Questionnaire (HAQ), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP),RF,and hemoglobin. The method of Pearson correlation or Spearman rank correlation was performed for assessing the association between RF and these indices separately, normally distributed data for Pearson correlation, nonnormally distributed data for Spearman rank correlation. MAIN OUTCOME MEASURES: Correlation of RF with above mentioned factors. RESULTS: None of the correlation coefficients between RF and indices including age,disease duration, functional status,radiographic change,joint pain, morning stiffness, TJC,TJS,SJC,SJS,HAQ,ESR,CRP,hemoglobin were significant (P＞0.05). CONCLUSION: No associations between RF and activity or severity of RA are studied.