BACKGROUND: Spinal cord can regenerate after injury in certain microenvironment. Olfactory ensheathing cells (OECs)have the characteristics of astrocytes and Schwann cells and can accelerate the spinal cord axonal regeneration.OBJECTIVE: To make injured thoracic cord rat models and observe the effect of OECs on injured spinal cord axonal regeneration.DESIGN: Observational experiment.SETTING: Second Affiliated Hospital of Sun Yat-Sen University.MATERIALS: The experiment was performed at the Second Affiliated Hospital of Sun Yat-Sen University from January 2001 to November 2002.Totally 20 adult SD male rats with the body mass of (380±20) g were provided by Experimental Animal Center of Sun Yat-Sen University (number of institution license SYXK2004-0020). There were DMEM culture solution with low glucose (L-DMEM, GibcoBRL), fetal calf serum (FCS) (Hyclone), myelin basic protein (MBP) (Sigma) and nerve growth factor receptor antibody (Sigma). They were divided into cell transplantation group and control group by the method of random digits table with 10 in each group.METHODS: The adult SD rats were anaesthetized and decapitated to obtain the whole olfactory bulb and then isolate olfactory nerve with a sterile operation. Thoracic cord injury models were established by modified Allen method. 10μL OECs suspension (2.5×1010 L-1) was injected into injured spinal cord of the cell transplantation group, whereas DMEM/F12 (1:1) culture solution of the same dose was injected in the control group. The influence of OECs on spinal cord axonal regeneration was observed by histological and immunohistochemical method 6 weeks after transplantation.MAIN OUTCOME MEASURES: ①OECs were identified by nerve growth factor receptor antibody staining. ②Repair of myelin sheath was observed by MBP staining. ③Nerve axonal regeneration was observed by argentaffin staining. RESULTS: Two rats in the cell transplantation group and 3 rats in the control group died, so totally 15 rats were involved in the result analysis. ①In the cell transplantation group,injured spinal meninges were integrated,but spinal cord became thin as compared with the normal spinal cord. By hematoxylin-eosin (HE) staining, multipolar cells appeared in injured region and the cells were fused excessively with myeloid tissues. It proved that survival OECs were integrated well within the host. New axons were clustered in bundles and infiltrated by small round lymphocytes. By argentaffin staining, regenerated axons grew in tissues of injured region, which mostly accompanied with fascicular-arranged multipolar cells. In the control group, spinal cord became thin markedly and spinal meninges were integrated. No new axon appeared in the injured spinal cord by HE staining. No regenerated axon appeared by argentaffin staining, either. ②In the cell transplantation group, most multipolar cells were clustered in bundles. A mass of positive granules of nerve growth factor receptor antibody appeared in cytoplasm, which further verified that OECs still survived and integrated well within the host 6 weeks after transplantation. Linear MBP positive fibers appeared in the injured region by MBP staining,which indicated that myelin-like substance appeared and drew closely in both ends of injured region. At the same time,MBP positive substance also appeared in the multipolar cells, which illustrated that transplanted OECs could induce the occurrence of myelin-like substance. No regenerated axon was found in the control group.CONCLUSION: Delayed transplantation of OECs can survive and induce the occurrence of myelin-like substance in injured spinal cord of adult rats.

Objective: Bilberry extract can inhibit the growth of cancer cells,but its effect on cervical carcinoma is rarely reported.This study was to explore the effects of bilberry extract on the expressions of Bcl-2,Bax,Fas and FasL in Hela cells.Methods: We determined the apoptosis of Hela cells by Hoechst33342/PI fluorescent staining and DNA ladder and Annexin V/PI double-staining after exposed to bilberry extract at the concentrations of 0.025-25 ?g/ml for 24 hours,and the protein expressions of Bcl-2,Bax,Fas and FasL were detected by Western blotting.Results: Obvious apoptosis-associated morphological changes and specific DNA ladder bands were observed in the Hela cells after exposed to bilberry extract at the concentrations of 0.025,0.25,2.5 and 25 ?g/ml,and the apoptosis rates of the cells were(4.13 ? 0.63)%,(5.41 ? 0.77)%,(8.74 ? 1.27)% and(12.05 ? 1.03)%,respectively.Western blot showed increased expressions of bax,Fas and FasL,and a decreased expression of bcl-2.Conclusion: Bilberry extract induces the apoptosis of Hela cells by up-regulating the protein expressions bax,Fas and FasL and down-regulating that of bcl-2.

Objective To investigate the clinical significance of serum interleukin (IL)-6,IL-8,tumor necrosis factor (TNF)- α, high-sensitivity C-reactive protein (hs-C RP), total immunoglobulin (Ig) and complement C levels in children with mycoplasma pneumoniae pneumonia (MPP). Methods Sixty-one cases of children with MPP (observation group) and 60 healthy children (control group) were selected. The serum IL-6 and IL-8 levels were tested by enzyme linked immunosorbent assay(ELISA) method; the serum TNF-α level was tested by γ radioimmunoassay (RIA) counter; the serum hs-CRP, total Ig and complement C levels were tested by rate nephelometry method. Results The serum IL-6 concentration was (109.31 ±54.72) ng/L, IL-8 concentration was (128.19 ±65.87) ng/L, hs-CRP concentration was (34.13 ± 19.21) mg/L, total Ig concentration was (12.62 ± 5.51) g/L and complement C concentration was (2.98 ± 0.97) g/L in the observation group, and they were higher than those in the control group [(67.23 ±32.18) ng/L, (31.78 ±8.91) ng/L, (1.62 ±1.34) g/L, (6.71 ±3.65) g/L, (1.46 ±0.45) g/L]. The differences were significant (P ＜ 0.05 or ＜ 0.01). The serum TNF- t level between two groups had no significant difference (P＞ 0.05). Conclusions The serum cytokines IL-6 and IL-8, hs-CRP, total Ig and complement C play an important role in MPP. It has important significance to detect the serum cytokines IL-6 and IL-8, hs-CRP, total Ig and complement C levels in children with MPP.

Objective To observe the effects of cryopreserved olfactory ensheathing cells(OECs )transplantation on spinal cord axonal regeneration and spinal func tional improvement.Methods Twenty-four rats were divided into experimental a nd control groups,with each group of 12rats.The spinal cord in-jury was es tablished by transecting the spinal cord with microsurgery scissors.OECs were p urified from SD rat olfactory bulb and cultured in DMEM and cryopreserved(-1 20℃)for 2weeks.OECs suspension[(1～1.4)?10 5 /ml ]was transplanted into transected spinal cord at T 10 level,while the DMEM solution was in ject-ed instead in the control group .At 6weeks and12weeks after transplantation,the rats were evaluated with cl imbing test and MEP monitoring.The samples of spinal cord were procured and st udied with histological and immunohistochemical measurement.Results At 6weeks after transplantation,all of the rats in both transplanted and control group were paraplegic,and MEPs could not be recorded.Morphol ogy of trans-plante d OECs was normal,and OECs were interfused with host well.Axons could regrow i nto gap tis sue be-tween the spinal cords.Both OECs and regrowed axons were im munoreactive for MBP.No re grown axons were found in the control group.At 12weeks after transplantation,2rats(2/7)had lower ex tremities mus-cle contraction,2rats(2/7)had hip and /or knee active movement,and MEP of 5rats(5/7)could be recorded in the calf in the transplantation group.Non e of the rates(7/7)in the control group had functional improvement,and n one of them had MEPs recorded.In the transplanted group,histological and imm unohis-tochemical methods showed the number of transplanted OECs reduced and s ome regrown axons had reached the end of transected spinal cord.However,no r egrown axons could be seen except scar forma tion in the control group.Concl usion Cryopreserved OECs could integrate with the host and promote re growing a x-ons across the transected spinal cord ends.

AIM: To investigate 1) the role of transforming growth factor-?_1(TGF-?_1) and macrophage infiltration during the development of myocardial fibrosis(MF) in rats after myocardial infarction(MI);and 2) mechanisms of MF post-MI and the inhibitory effect of angelica.METHODS: Sprague-Dawley(SD) rats were subjected to MI by ligating the left anterior descending coronary artery.The animals were randomly divided into three groups: sham,MI and MI+angelica.After 24 hours of ligation,rats received angelica(20 mL?kg~(-1)?d~(-1),ip) or saline.Left ventricular hemodynamics were measured and rats were killed at week 1,week 2 and week 4, respectively.Collagen content,macrophage infiltration and TGF-?_1 expression were examined in the non-infarcted area.RESULTS: ① In MI group,the numbers of macrophage and TGF-?_1 expression were significantly upregulated compared to sham at week 1 post-MI and remained elevated at week 4(P

BACKGROUND:Olfactory ensheathing cells (OECs) have been shown to possess the potential of repairing injured spinal cord, but their biological characteristics after transplantation in vivo are not well understood.OBJECTIVE: To investigate the migration of OECs after transplantation into the injured spinal cord of adult rats.DESIGN: Randomized and controlled experiment.SETTING: Department of Orthopedics, Guangdong Provincial People's Hospital; Experimental Animal Center of the North Campus of Sun Yat-sen UniversityMATERIALS: Totally 38 2-month-old male SD rats with body mass of (350 ±20) g were used in this study.METHODS: This experiment was conducted in the Experimental Animal Center, North Campus of Sun Yat-sen University between February 2004and May 2004. Two SD rats were used to extract the OECs, which were stained with Hoechst 33342. Totally 36 SD rats were subsequently randomized into 3 groups, namely rostral transplantation group, caudal transplantation group and control group with 12 rats in each group. The rats in the rostral and caudal transplantation groups subjected to operations to establish thoracic spinal cord injury model and OEC suspension was injected; in the control group, the rats were spared of thoracic spinal cord injury with only OEC suspension injection.MAIN OUTCOME MEASURES: Distribution of OECs in the spinal cord was observed under fluorescence microscope 1, 2, 4 and 6 weeks after operation, respectively.RESULTS: Of the rats in the 3 groups, 1 died in the rostral group, and 2in each of the caudal transplantation and control groups, leaving 29 rats for result analysis. The OECs in the rostral and caudal transplantation groups migrated longitudinally along the long axis of the spinal cord to a farthest distance of 8 mm and penetrating the scar tissues, but very few cells could reach the contralateral side. The OECs in the control group diffused locally without migration.CONCLUSION: OECs mainly migrate along the axons in white matter of the injured spinal cord, and their rostral and caudal migration does not differ in speed or amount. Only a small amout of OECs can across the transected gap of the spinal cord.

Objective: To investigate the effect of endocannabinoid system on cardiac hypertrophy in experimental rats with chronic intermittent hypoxia and to study the impact of endocannabinoid antagonist, rimonabant in such pathological processing.
Methods: A total of 48 male Wistar rats were divided into 6 groups. 4 and 6 weeks of Normal control group, 4 and 6 weeks of Hypoxia group, 4 and 6 weeks of Hypoxia with rimonabant intervention group. n=8 in each group. The rats were sacrificed to measure left ventricular mass index (LVMI), the myocardial cell morphological changes were observed by optical microscope, the expression of cardiac calcium/calmodulin-dependent protein kinase II (CaMKII) and cardiotrophin-1 (CT-1) were detected by immunohistochemistry at 4 and 6 weeks respectively.
Results: Compared with 4 and 6 weeks of Normal control group, the LVMI, cardiac hypertrophy condition, CaMKII and CT-1 were increased in 4 and 6 weeks of Hypoxia group, all P<0.001. Compared with Normal control group and Hypoxia group, the above indexes in Hypoxia with rimonabant intervention group were higher than those in Normal control group while lower than those in Hypoxia group, all P<0.001.
Conclusion: The Chronic intermittent hypoxia could induce myocardial hypertrophy via endocannabinoid system disorders, such pathological processing could be reduced by rimonabant intervention.

Objective To explore the correlation between the reliability and validity of Brunel balance assessment(BBA)with the activities of daily living(ADL).Methods The BBA,ADL and walking ability test were performed on 215 patients conforming to the inclusion standard by two professionals.The Pearson correlation analysis,factor analysis and descriptive statistical analysis were adopted.Results The BBA total score and Cronbach's α coefficient in 3 dimensions was 0.852～0.941(P＜0.01),moreover the relative coefficient of each item with its dimension was higher that that of other dimensions;3 common factors were extracted by the factor analysis,the contribution rate was 83.916％;but BBA had the ceiling effect,the relative coefficient of BBA with ADL and walking ability was 0.284-0.709(P＜0.05).Conclusion BBA has a good reliability and validity and can better reflect the functions of ADL and walk.

Objective To investigate the effects of modafinil,a new wake-promoting agent,on simulated flight performance during 48 h sleep deprivation(SD).Method Six male healthy young volunteers were exposed to two periods of 48 h continuous wakefulness during the crossover experiment.In one period,three 200 mg doses of modafinil were given and in the other,separated by two weeks,matching placebos were administered.The SD time started from 8:00 of the first day to 8:00 of the third day.Drug was given at 0:00,16:00 of the second day and 0:00 of the third day.Flight performance in J7-E flight simulator was tested at 21:00 of the first day and 1,3,5,7 h after each drug administration.Result The scores of simulated flight performance in the placebo group decreased with time during SD and became significant at 1:00～7:00 of the third day.Numbers of errors increased with time during SD,especially in the left ascending turn stage and the descending & landing stages.Compared with the placebo,flight performance was significantly increased after the third administration of modafinil.Numbers of errors during 48 h SD and at 1:00～7:00 of the third day in modafinil group was 19% and 40% lower than those in placebo group respectively.Conclusion Simulated flight performance during 48 h SD is significantly improved after repeated administration of modafinil,especially when the impacts of SD and the circadian trough are combined.

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