AIM: To determine tanshinone Ⅱ_A and salvianolic acid B by HPLC gradient elution. METHODS: HPLC was used with Hypersil-ODS C_(18) column(150 mm?4.6 mm,5 ?m).The mobile phase consisted of methanol-water 0~20 min(35-90:6510),20-25 min(90-95:10-5),25-30 min(95:5)gradient elution.The flow rate was 1.0 mL/min with column temperature at 30 ℃.The UV detection wavelength was at 286 nm in 0-20 min and 270 nm in 20-30 min,respectively. RESULTS: The linear range of tanshinone Ⅱ_A and salvianolic acid B were in the range of 0.014 448-0.096 32(r=0.999 97), and 0.198 6-3.972 ?g(r=0.999 94),respectively.The average recoveries were 99.62%(RSD=2.28%) and 100.58%(RSD=2.5%),respectively.(CONCLUSION:) The method is accurate、sensitive、reproducible,and suitable for the quality control of Chaidan Jieyu Granules.

OBJECTIVE:To improve the writing quality prescriptions of traditional Chinese medicines and to facilitate the standardization of traditional Chinese medicine prescriptions.METHODS:A total of 15 000 prescriptions were sampled in our hospital in 2006 for an analysis of the problems in accordance with the related standards specified in Chinese Pharmacopoeia(CP,2005 edition)and the new "Prescription management method".RESULTS:The problems manifested as nonstandard in drug name and footnotes,or overdosage and so on.CONCLUSION:We should strengthen the management of the traditional Chinese drugs and improve our pharmaceutical care.

The expression of Survivin, CDK4 and Ki-67 and the clinical significance in pediatric acute leukemia (AL) were investigated. The expression of Survivin, CDK4 and Ki-67 proteins was detected by using immunohistochemical assay in 37 children with AL and 10 children with normal bone marrow as controls. The positive expression rate of Survivin, CDK4 and Ki-67 was 45.9 %, 56.8 %, and 40.5 % respectively in 37 AL children, which was significantly higher than in control group accordingly (P<0.05). The expression of Survivin was positively correlated with CDK4 (P=0.007) and Ki-67 (P=0.008). In conclusion, all Survivin, CDK4 and Ki-67 proteins are over-expressed in pediatric AL and involved in the modulation of apoptosis and proliferation in pediatric AL.

To assess the relationship between pronuclear scoring and day-3 embryo quality and pregnancy outcome and to determine the clinical value of pronuclear stage scoring system in human in vitro fertilization-embryo transfer (IVF-ET) program, a pronuclear scoring system was used to score zygotes 16-20 h after insemination during conventional IVF or intracytoplasmic sperm injection (ICSI). The embryos were classified into groups Z1, Z2, Z3 and Z4. Comparisons were made of the rates of arrested embryos and excellent embryos on day 3. Comparisons of pregnancy outcome were made only in those patients in whom cohorts of similarly Z-scored embryos were transferred. The results showed that there were less arrested embryos and more excellent embryos on day 3 in groups Z1 and Z2 than those in group Z3 and Z4. More embryos arrested and less excellent embryos developed in group Z4 than group Z3. The clinical pregnancy rates resulting from the transfer of single pronuclear score homologous embryo types were similar among groups Z1, Z2 and Z3. Implantation rates of group Z1 were higher (P<0.05) than that of group Z3. These findings suggests that pronuclear scoring can predict developmental ability on day 3 and implantation potential. A evaluation that combines the Z-score and day 3 embryo morphology is useful in the determination of the most viable embryos and the number of embryos for transfer.
Cell Nucleus/*metabolism ; Embryo Implantation ; Embryo Transfer/*methods ; Fertilization in Vitro ; Infertility/therapy ; Models, Biological ; Oocytes/metabolism ; Ovary/*metabolism ; Pregnancy Outcome ; Spermatozoa/metabolism

Objective To study the mechanism of marcosomia by investigating insulin-like growth factor 2(IGF_2)imprinting status,expression level and the promoter usage in the placenta of macrosomia. Methods We selected heterozygous cases for Apa Ⅰ polymorphism in exon 9 of IGF_2 gene and then analyzed its imprinting status in 168 placentas of macrosomia and normal pregnancies.IGF_2 transcription levels and promoter usages in macrosomic and normal placenta were evaluated by using semi-quantitative RT- PCR assay.Results Thirty specimens of macrosomic placenta and 30 of normal placenta were identified as heterozygous for IGF_2.All of the heterozygous specimens showed maintenance of imprinting.The expression of placental IGF_2 mRNA(2.2?1.2)was significantly higher in macrosomia than that of normal weight group (1.6?0.6,P 0.05).Conclusion It is possible that over expression of IGF_2 in placenta contributes to macrosomia while the promoter usage and imprinting status are not associated with macrosomia.

OBJECTIVETo investigate the influences of experimental osteoporosis (OP) on bone healing of autologous iliac crest graft around dental implants in rabbits.

METHODSTwenty Japanese rabbits were randomly divided into two groups. Bilaterally ovariectomy was performed on experimental group and control group received sham-operation. Twelve weeks later, femoral bones were examined for bone mineral density (BMD) to verify OP status. Then bone defects were made in the proximal metaphysis of the tibiae and autologous iliac crest grafts with simultaneous implant placement were performed. The animals were killed at 8 and 12 weeks after bone graft surgery. Undecalcified sections were prepared and examined histologically and histomorphometrically.

RESULTSOsteoporotic status caused by ovariectomy was verified by significantly decreased BMD in experimental group (P < 0.001). At 8 and 12 weeks after bone graft surgery, osseointegration was observed in both groups. However, thickness of cortical bone (TCB), bone volume in cancellous area (BVC), implant-bone contact rate (IBCR) at bone graft area all significantly decreased in experimental group when compared with control group (P < 0.01). Newly formed bone was also less in experimental group than that in control group.

CONCLUSIONAlthough experimental OP may not delay osseointegration of dental implants in autologous iliac crest graft, it certainly promotes resorption of bone grafts, decreases cancellous bone volume and implant-bone contact rate. Therefore it may be an important risk factor for patients receiving autologous bone graft with simultaneous implant placement.

Animals ; Bone Transplantation ; Dental Implants ; Female ; Ilium ; transplantation ; Osseointegration ; Osteoporosis ; pathology ; Ovariectomy ; Rabbits

BACKGROUNDThe urokinase plasminogen activator system is believed to play an important role in degradation of the extracellular matrix associated with cartilage and bone destruction; however its precise roles in temporomandibular disorders have not yet been clarified. The aims of this study were to investigate the gene expression of fibrinolytic factors urokinase plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) in the articular cartilage of rabbit temporomandibular joint (TMJ) with disc displacement (DD) and to probe the relationship between fibrinolytic activity and cartilage remodeling.

METHODSDisc displacement of right joints was performed in 36 of 78 rabbits under investigation. The animals were sacrificed at 4 days and 1, 2, 4, 8 and 12 weeks after surgery, respectively. The right joints of these animals were harvested and processed for the examination of mRNA expression of uPA and PAI-1 in articular cartilage using in situ hybridization techniques.

RESULTSThe expression of uPA and PAI-1 was co-expressed weakly in the chondrocytes from transitive zone to hypertrophic zone and mineralized zone, while no hybridizing signals were shown in proliferative zone and superficial zone in control rabbits. The most striking was the up-regulation of uPA and PAI-1 mRNA in 4-day rabbits postoperatively at the onset of cartilage degeneration. The strongest hybridizing signals for uPA and PAI-1 were seen in 2-week rabbits postoperatively. After 2 weeks, the expression of uPA and PAI-1 began to decrease and reached nearly normal level at 12 weeks.

CONCLUSIONSThe expression of the uPA/PAI-1 system coincides with the pathological changes in condylar cartilage after DD. The uPA/PAI-1 system may be one of the essential mediators in articular cartilage remodeling.

Animals ; Cartilage, Articular ; metabolism ; Female ; Joint Dislocations ; metabolism ; pathology ; Male ; Mandibular Condyle ; metabolism ; pathology ; Plasminogen Activator Inhibitor 1 ; genetics ; RNA, Messenger ; analysis ; Rabbits ; Temporomandibular Joint ; metabolism ; Temporomandibular Joint Disc ; Urokinase-Type Plasminogen Activator ; genetics

OBJECTIVETo investigate the effect of anterior disc displacement on the expression of urokinase plasminogen activator and its inhibitor-1 (uPA/PAI-1) in synovial tissues.

METHODSForty Japanese white rabbits were used in this study. The animals were killed at 4 days, 1, 2, 4, 8 and 12 weeks postoperatively, respectively. In situ hybridization technology was applied to detect the expression of uPA/PAI-1 mRNA in synovial membrane.

RESULTSIn normal synovial tissues, synovial lining cells and a few fibrosblasts with mild positive staining were occasionally seen. More synovial lining cells and fibrosblasts with moderate postive signals were found 1 week after operation. Since then, the degree of staining for uPA/PAI-1 increased gradually. By the end of 12 weeks postoperatively, strong signals of uPA/PAI-1 mRNA were detected.

CONCLUSIONThere is a harmonized uPA/PAI-1 system existing in synovial tissues. The high expression of uPA and PAI-1 mRNA in synovial tissues indicates that the uPA/PAI-1 system may play an important role in the process of synovitis resulted from anterior disc displacement.

Animals ; In Situ Hybridization ; Plasminogen ; Plasminogen Activator Inhibitor 1 ; RNA, Messenger ; Rabbits ; Synovial Membrane ; Urokinase-Type Plasminogen Activator

BACKGROUNDAccurate knowledge of the spinal structural functions is critical to understand the biomechanical factors that affect spinal pathology. Many studies have investigated the human vertebral motion both in vitro and in vivo. However, determination of in vivo motion of the vertebrae under physiologic loading conditions remains a challenge in biomedical engineering because of the limitations of current technology and the complicated anatomy of the spine.

METHODSFor in vitro validation, a human lumbar specimen was imbedded with steel beads and moved to a known distance by an universal testing machine (UTM). The dual fluoroscopic system was used to capture the spine motion and reproduce the moving distance. For in vivo validation, a living subject moved the spine in various positions while bearing weight. The fluoroscopes were used to reproduce the in vivo spine positions 5 times. The standard deviations in translation and orientation of the five measurements were used to evaluate the repeatability of technique. The accuracy of vertebral outline matching with metallic marks matching technology was compared.

RESULTSThe translation positions of the human lumbar specimen could be determined with a mean accuracy less than 0.35 mm and a mean repeatability 0.36 mm for the image matching technique. The repeatability of the method in reproducing in vivo human spine six degrees of freedom (6DOF) kinematics was less than 0.43 mm in translation and less than 0.65° in rotation. The accuracy of metallic marks and vertebral outline matching did not show significant difference.

CONCLUSIONSCombining a dual fluoroscopic and computerized tomography imaging technique was accurate and reproduceable for noninvasive measurement of spine vertebral motion. The vertebral outline matching technique could be a useful technique for matching of vertebral positions and orientations which can evaluate and improve the efficacy of the various surgical treatments.

Biomechanical Phenomena ; Fluoroscopy ; methods ; Humans ; In Vitro Techniques ; Lumbar Vertebrae ; anatomy & histology ; physiology ; Middle Aged ; Spine ; anatomy & histology ; physiology

Objective:To summarize the clinical features and pathological changes of peripheral tissues from patients with neuronal intranuclear inclusion disease (NIID) diagnosed by genetic tests.Methods:Repeat-primed polymerase chain reaction was used to confirm the GGC repeated expansion in the 5′ untranslated region of the NOTCH2NLC gene in patients with suspected NIID who had visited the Department of Neurology of Peking University First Hospital from January 2018 to February 2020. The clinical data and pathological changes of peripheral tissues from patients with genetically diagnosed NIID were collected retrospectively and analysed. Immunostaining with anti-p62 and anti-ubiquitin antibody was performed on peripheral biopsy specimens.Results:Totally nine patients with NIID who had GGC repeated expansion in the NOTCH2NLC gene were found. Five patients were familial (from three faimilies), and four patients were sporadic. The age of onset was 36-61(51.33±7.12) years. The most common symptoms in this NIID group were episodic emotion and personality change (8/9), paroxysmal disturbance of consciousness (6/9) and intermitant head discomfort (6/9). Other symptoms included cognitive dysfunction, limb weakness, limb sensory disturbance, bladder dysfunction, ataxia, seizures and psychiatric symptoms. Brain magnetic resonance imaging showed high signals along the corticomedullary junction on diffusion-weighted image in eight out of nine patients. Skin biopsied samples from nine patients demonstrated the presence of eosinophilic intranuclear inclusions (IIs), appearing in the nucleus of fibroblasts, fat cells and ductal epithelial cells of sweat glands on hematoxylin-eosin staining. IIs were positive on anti-p62 and anti-ubiquitin immunostaining. Electron microscopy indicated the IIs were composed of a pile of filament materials without membrane. Muscle biopsies from two patients showed no obvious neurogenic or myogenic pathologic changes, except in one patient several rimmed vacuoles fibers were found. In one patient sural nerve biopsy showed severe demyelinating pathological changes. No IIs were found in the muscles and peripheral nerve tissue either by histological examination or by immunohistochemical staining with anti-p62 or anti-ubiquitin, while IIs were found by immunofluorescence staining with both anti-p62 and anti-ubiquitin in three patient′s tissue. Conclusions:The phenotype of this NIID patient group is adult-onset NIID, with episodic encephalopathy as the main clinical manifestation. Skin biopsy has high pathological diagnostic value for NIID. The immunofluorescence staining with anti-p62 and anti-ubiquitin is easier to detect the presence of IIs than histological staining and immumohistochemical staining.