BACKGROUND:Compared with traditional autogenous bone graft, composite, as the carrier of seed cells, possesses advantages of fewer traumas and no limitation of donor site in repairing bone defect.OBJECTIVE: To observe the ability of the composite of β-tricalcium phosphate artificial bone-hyaluronic acid-type I collagen (β-TCP/HA/COL-I), as induced bone marrow stromal cell (MSC) carrier, to repair rabbit radial defect, and the feasibility with the composite as bone substitute material.DESIGN: A randomized and controlled trial.SETTING: Department of Orthopaedics, Renmin HospitaL, Wuhan University.MATERIALS; The study was conducted in the Department of Orthopaedics Renmin Hospital, Wuhan University between September 2003 and July 2004. Thirty-one New Zealand big white rabbits, aged 6 months, with body mass of 1.5 to 2.0 kg were enrolled in this study. The rabbits were randomized into control group (n =4) and experimental group (n =27).METHODS : ①In vitro induction and culture of MSCs was performed on 31 white rabbits, and the alkaline phosphatase (ALP) positive ratio of induced MSCs was observed. The structure of β-TCP/HA/COL-I was observed under scanning electron microscope. ② A 2 cm radial defect was created through operation. Eight weeks later, composites of β-TCP/HA/COL-I/MSCs were implanted into one side of rabbits in the experimental group, and autogenous bone was implanted into the other side. Rabbits in the control group were untouched. ③All the animals in the experimental group were randomly sacrificed at postoperative 4,8 and 12 weeks, 6 rabbits at 4 and 8 weeks, 15 at 12 weeks; Animals in the control group were sacrificed at 12 weeks. Gross observation, X-ray photographing, haematoxylin-eosin (HE) dyeing, and assessment of inorganic ingredient were performed. Osteogenic area and biomechanical tests were performed at 12weeks. Repairing effects on bone defect in each group were compared.MAIN OUTCOME MEASURES: The ALP positive ratio of induced MSCs; The structure of composite ofβ-TCP/HA/COL-I;Gross observation; X-ray photographing; HE dyeing and assessment of inorganic ingredient; Osteogenic area and biomechanical tests.RESULTS: All the 31 rabbits entered the stage of result analysis. ① The ALP positive ratio of cells reached 75% after induction and culture. ② Scanning electron microscope observation showed that 3 kinds of materials with abundant cellular structure distributed evenly. ③ The osteogenic area at 12 weeks was (72.5±3.6)% and (76.7±6.2)% in the experimental group and autogenous group, respectively (P ＞ 0.05). ④The maximum bending moment was (521.0±61.1) and (554.3±53.3)N·mm in the experimental group and control group, respectively; The maximum displacement at point of application of force was (0.816±0.071)and (0.870±0.103)respectively, without significant difference (P ＞ 0.05). ⑤Inorganic ingredient in the composite was 75%, 57% and 42% at 4,8 and 12 weeks respectively, suggesting that the inorganic component in the material was gradually decomposed with the elongation of time. ⑥Results of gross observation,X-ray photographing, histopathological examination, biomechanical test showed that with the elongation of time, composite of β-TCP/HA/COL-I/MSCs could repair bone defect in the experimental group, while bone defect in the control group had not been repaired.CONCLUSTON: Composite of β-TCP/HA/COL-I /MSCs possesses the same effect on repairing bone defect as autogenous bone, so it may be used as autogenous bone graft substitute.

This article was aimed to evaluate the α-glucosidase inhibitory activity of extracts from the root of Salvia miltiorrhiza Bunge and different processed products. The α-glucosidase inhibitory activity was screened with acarbose as positive control and by α-glucosidase inhibitory model in v itro . The results showed that the ex-tracts from the root of S. miltiorrhiza and different processed products had inhibitory activity. And the activity was higher than that of acarbose. In addition to the MeOH extract of S. miltiorrhiza carbon, the inhibitory activity of MeOH extracts from other processed products were higher than that of MeOH extract of S. miltiorrhiza. The in-hibitory activity of petroleum ether extracts of different processed products were close to S. miltiorrhiza. EtOAC extracts were lower than that of S. miltiorrhiza. The n-BuOH extracts were higher than that of S. miltiorrhiza. The inhibitory activity of extracts was positively correlated with concentrations, and it depended on the concentra-tion. It was concluded that the processed products of S. miltiorrhiza can strengthen α-glucosidase inhibitory activ-ity in different degrees.

This study was aimed to assay the antibacterial activity of compounds from Rabdosia rubescens. Disc dif-fusion (K-B method) was used to screen the in vitro antibacterial activity. All kinds of chromatography were used to isolate the chemical constituent and structure was identified by MS and NMR spectroscopy. The results showed that six compounds were isolated and identified as oridonin (1), rosmarinic acid (2), caffeic acid (3), salicylic acid (4), ferulic acid (5), vanillic acid (6). Oridonin had activity against Staphylococcus aureus (SA), Methcillin-resistant Staphylococcus aureus (MRSA), β-lactamase positive Staphylococcus aureus (ESBLs-SA), and showed the highest ac-tivity (MIC is 3.125, 6.25 and 6.25 μg·disc-1, respectively), but was still weaker than that of berberine as positive control (MIC is 0.156 μg·disc-1). Ferulic acid had activity against SA and MRSA (MIC is 50 and 50 μg·disc-1, re-spectively). Salicylic acid had only activity against SA (MIC 50 μg·disc-1). It was concluded that oridonin, ferulic acid and salicylic acid were the main antibacterial activity compounds from R. rubescens.

Objective To compare the effects of propofol versus isoflurane on brainstem auditory evoked potential (BAEP) and explore the difference in the effects of the two anesthetics on the brainstem. Methods Thirty ASA Ⅰ or Ⅱ patients aged 20-50 yr without heating disorder, scheduled for elective surgery performed under general anesthesia were randomly divided into 2 groups (n = 15 each): propofol group (group P) and isoflurane group (group Ⅰ). SpO2, PET CO2, BIS and BAEP were continuously monitored before and during anesthesia. Anesthesia was induced by propofol administered by TCI or isoflurane inhalation. Tracheal intubation was facilitated with vecuronium. The patients were mechanically ventilated. PET CO2 was maintained at 35-40 mm Hg and SpO2 at 98%-100%. After intubation BIS was maintained at 70 and 50 respectively ,the latency of the wave Ⅰ , Ⅱ and Ⅴ and the inter-peak latency (IPL) betwecn wave Ⅰ -Ⅲ , Ⅲ-Ⅴ and Ⅰ -Ⅴ were recorded.Results In group P there was no significant difference in the latency of the wave Ⅰ , Ⅲ and Ⅴ and the IPL between wave Ⅰ - Ⅲ , Ⅲ - Ⅴ and Ⅰ - Ⅴ between the baseline before anesthesia and at BIS 70 and 50. In group Ⅰ the latency of wave Ⅲ and Ⅴ and the IPL between wave Ⅰ - Ⅲ , Ⅲ - Ⅴ and Ⅰ - Ⅴ were significantly longer at BIS 50 than the baseline before anesthesia, while the latency of wave Ⅲ and Ⅴ and the IPL between wave Ⅰ -Ⅲ andⅠ -Ⅴ at BIS 50 were significantly longer than that at BIS 70. At BIS 50 the latency of wave Ⅴ and the IPL between wave Ⅰ -Ⅴ were significantly longer in group Ⅰ than in group P. Conclusion At comparable depth of anesthesia propofol exerts less depressant effects on BAEP indicating less depression of brainstem.

Objective To Investigate the influence of high-intensity pulsed electromagnetic field (HIPEMF) on neural differentiation of neonatal rats neural stem cells in vitro.Methods Neural stem cells (NSCs) were isolated from the subventricular zone of 3-day-old neonatal rats and cultured with serum-free condition medium for 14 days.All the NSCs were then randomly classified into an experimental group which received the stimulation of HIPEMF (0.1 Hz,4 T,8 pulses) and a control group which received no special intervention.Differentiation of the culture was induced by addition of 10％ fetal bovine serum on the first day after intervention,the morphological changes of cells were observed under the microscope at different time points.The NSCs adhered to the wall and differentiated for seven days,the immunofluorescence was employed to observed and calculate the ratio of differentiated cells with astrocyte marker GFAP or neuronal markers TUJ1.RT-PCR and western blotting were used to measure the expression levels of the differentiated cells based on gene and protein levels,respectively.Results Immunofluorescence staining showed the number of TUJI positive cells in the experimental group(33.4％ ± 5.1％)was significantly more than the control group (26.5％ ± 7.0％),while the number of GFAP positive cells was decreased(23.9％ ± 5.0％) as compared with the control group(36.2％ ± 2.2％).RT-PCR showed that the TUJ1 mRNA expression levels in the experimental group was (1.682 ± 0.086) times of the control group.Western blot showed that the expression of TUJ1 (0.729 ±0.061) in the experimental group was higher than in the control group (0.590 ± 0.157),while the expression of GFAP in the experimental group (0.566 ± 0.056) was less than in the control group(1.034 ± 0.051).Conclusions HIPEMF facilitates differentiation of neural stem cells to neurons,at the cost of reducing astrocytic differentiation.

Objective: To investigate the multi-imaging diagnostic values, especially MSCT technology in patients with congenital aortic diverticulum with its clinical application. Methods: The MSCT ifndings in 12 patients with congenital aortic diverticulum were retrospectively analyzed. Results: There were 9 patients with right aortic arch and 1 with left aortic arch, all of them having coexisted aberrant subclavian artery which initially dilated like aneurysm by diverticulum changing (Kommerell diverticulum), and there was 1 patient with incomplete double aortic arch with atresia of left arch combining retro-esophageal aortic diverticulum (RAD) and 1 patient with ducts diverticulum. Echocardiogram only made the suggestive diagnosis of speeding up blood lfow or right aortic arch in 4 patients. While MSCT accurately displayed the diverticulum for the location, morphology and with or without other complications. The post-eroanterior chest radiograph indicated “double aortic node” as the special sign in 8 patients. The echocardiogram, X-ray and MSCT for correctly diagnosing the aortic diverticulum were as 0, 72.7% and 100% respectively. Conclusion: MSCT is a rather ideal non-invasive diagnosing method for aortic diverticulum, meanwhile X-ray could also make suggestive diagnosis; if MSCT and X-ray joint with echocardiogram examination may provide the effective supplement for valve structure and hemodynamics condition in relevant patients.

Objective To analyze the relationship between polymorphisms of EDNRB gene and Hubei provincial patients of Han ethnicity with sporadic Hirschsprung disease(sHD). Methods Peripheral blood samples from 104 patients with sHD and 84 parents of 42 patients, and 120 normal children(as controls) were collected. PCR-SSCP and direct DNA sequencing were used to detect mutations and polymorphisms of exon-4 in EDNRB gene. The differences of allele frequencies and genotype distribution in polymorphic sites were further analyzed between the three groups. Allele frequencies of SNPs in forty-two sHD trios were analyzed by transmission disequilibrium test(TDT), and the association between phenotype of HD and SNPs was analyzed. Results No mutant site was detected and one polymorphic site of c831 G→A(L277L) was observed in Hubei provincial patients of Han ethnicity with sHD. The allele frequency of A(68% vs 53%) and genotype frequency of AA(49% vs 30%) were significantly higher in sHD group than that in control group(P

BACKGROUND: Endoscopic transnasal lacrimal duct reconstruction by grafting of autogenous tissue is a novel method for treatment of severe lacrimal duct obstruction and it needs detailed anatomical data for surgery.OBJECTIVE: To study the applied microsurgical anatomy of lacrimal duct and to provide anatomical evidence for endoscopic transnasal lacrimal duct reconstruction by grafting of autoganous tissue.DESIGN, TIME AND SETTING: This study was performed at the laboratory of the Department of Ophthalmology, Armed Police General Hospital from July 2006 to June 2007.MATERIALS: Twenty 10% formaldehyde-treated adult cadaveric heads, 14 males and 6 females, comprising 40 lacrimal ducts were included in this study.METHODS: The cadaveric heads were split on the level of the line between the superior border of the superciliary arch and the site 10 mm higher than occipital tuberosity. After removal of brain tissue,the heads were decalcified for approximate 1 week with 10%nitric acid. This promised non-alteration of morphological structure and facilitation for surgical cutting. Following dissection of facial cranium in the median sagittal plane, the nasal septum was excised to expose the lateral wall of the nasal cavity.MAIN OUTCOME MEASURES: The anteroposterior diameter and depth of lacrimal fossa; at middle third level, the thickness of lacrimal fossa at the anterior lacrimal crest, vertical middle line, and posterior lacrimal crest; the cross section area of nasolacrimal canal upper opening, middle part, and lower opening; horizontal distance, 30° oblique distance, and 45°oblique distance from lacrimal caruncie to nasal cavity; distance from lacrimal caruncle to nasolacrimal canal upper opening; and the included angle between lacrimal caruncle-nasolacrimal canal upper opening line and Aeby's plane.RESULTS: The length, anteroposterior diameter, and depth of lacrimal fossa were (17.85±1.72) mm, (6.74+1.28) mm, and (3.09+0.78) mm, respectively. At middle third level, the thickness of lacrimal fossa at the anterior lacrimal crest,perpendicular bisector, and posterior lacrimal crest was (4.03±0.89) mm, (0.61±0.36) mm, and (0.63±0.24) mm, respectively.Anterior lacrimal crest was significantly thicker than vertical middle line and posterior lacrimal crest (P ＞ 0.05). Horizontal distance, 30°oblique distance, and 45° oblique distance from lacrimal caruncle to nasal cavity was (17.23±0.70) mm,(14.51±1.72) mm, and (17.34±2.38) mm, accordingly, with a difference which was not significant (P ＞ 0.05). The distance from lacrimal caruncle to lateral wall middle point of nasolacrimal duct superior opening was (11.86±1.84) mm, and the included angle between lacrimal caruncle-lateral wall middle point of nasolacrimal duct superior opening line and Aeby's plane averaged (49.9±1.8)°.CONCLUSION: The distances from lacrimal caruncle to nasal cavity and lacrimal sac and the included angles between lacrimal caruncle-nasolacrimal canal upper opening line and Aeby's plane provide guidance significance for selection of bony opening position on the lateral wall of nasal cavity and determinations of tunnel oblique angle and autogenous tissue length. Creation of bony tunnel should start from the middle or posterior middle part of lacrimal fossa and then extend towards anterior inferior region with an optimal downward oblique angle of 45°. The length of autogenous tissue used for lacrimal duct reconstruction should exceed 21.22 mm.

OBJECTIVETo study the influence of conditioned medium of rat brain microvascular endothelial cells on mitochondrial function of cortical neurons and the protective effect of Tongluo Jiunao Injection (TJI) on it.

METHODSFour kinds of conditioned endothelial cell (EC) cultured medium were prepared, i.e. the N-CM medium prepared by EC cultured in the normal conditioned medium without any treatment; the NT-CM prepared by EC cultured in N-CM and treated with TJI 1 microl/ml for 10 h; the I-CM prepared by EC cultured in the non-glucose kreb medium under hypoxia condition; and the IT-CM by EC pre-treatce with TJI 1 microl/ml for 4 h and cultured as that of I-CM. The levels of neuronic mitochondrial activity, membrane potential (MMP) and cytochrome C (Cyt C) were determined before and after the glucose-oxygen deprived model neurons of brain cortex being cultured with different kinds of conditioned EC cultured medium for assessing the effects of these media on mitochondria of injured neuron.

RESULTSAs compared with those of the normal neuron, the mitochondrial activity and MMP of all injured neurons decreased and Cyt C level increased significantly. But comparison of these indexes among neurons cultured with different conditioned EC culture media showed that the greatest extent abnormality revealed in the N-CM cultured neurons, which even greater than that in the model neuron; while that was less in the N-CM cutured neuron than in model neuron; as for those cultured in the NT-CM and IT-CM, i.e. the TJI treated cuture medium, the abnormal changes were reduced significantly when compared with those cultured in medium untreated with TJI (N-CM and I-CM), respectively (all P < 0.05).

CONCLUSIONThe paracrine secretion of the brain microvascular endothelial cells has evident regulatory effect on survival of the injured neurons, which might possibly be related to its protective effect on neuron mitochondrial function, and TJI could enhance the protective effect.

Animals ; Capillaries ; cytology ; Cells, Cultured ; Cerebral Cortex ; blood supply ; cytology ; Culture Media, Conditioned ; pharmacology ; Cytochromes c ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; cytology ; drug effects ; ultrastructure ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria ; drug effects ; metabolism ; physiology ; Neurons ; cytology ; drug effects ; ultrastructure ; Rats