Objective To explore the effect of hypoxic training on the barrier function of intestinal mucous membrane and underlying mechanism in rats.Methods Eighty 5-week-old male Sprague-Dawley rats were randomly divided into a control group,an exercise training group,a hypoxia control group and a hypoxia exercise group,each of 20.The altitude training was conducted through swimming training in artificial hypoxia environment.Two and 6 weeks after the intervention,the tissue structure and ultrastructure of small intestine mucosa were observed.The content of diamine oxidase (DAO)and D-lactate (D-LA),tumor necrosis factor-α (TNF-α)and nuclear factor κB (NF-κB)in plasma and the mRNA level of occludin in ileal tissue were measured.Results (1)Electron microscopy data showed that,after six weeks,compared with the control group,the microvilli of rats in the exercise training group were sparser and arranged irregularly.Furthermore,the gap between epithelial cells became wider.In addition,the number of mitochondria decreased significantly and cristae were vague.For the hypoxia control group,the microvilli shortened significantly and arranged irregularly.Moreover,the gap between cells became wider with partial denatured mitochondria.For the Hypoxia exercise group,the number of mucosal epithelium microvilli in the bowels reduced significantly and the microvilli shortened significantly.Similar to that of the hypoxia control group,the gap between epithelium cells growed wider.However,the cellular structure were fuzzier,and the denature of mitochondria worsened,with the cristae being vague even disappearing partially.(2)Two weeks of exercise training reduced the number of intestinal microvilli significantly (P＜0.01),but increased the plasma level of the DAO and D-LA,as well as the expression level of NF-κB in intestinal tissue significantly (P＜0.05).Hypoxic exposure significantly reduced the mRNA level of oceludin in small intestine (P＜0.01),but significantly increased the plasma level of DAO and D-LA (P＜0.05 vs.control)and the expression of TNF-α and NF-κB in small intestine (P＜0.01).There was no significant interaction between two weeks of exercise training and hypoxia exposure either on the reduction of the number and height of intestinal microvilli,or the transcription level of occluding in small intestine,or the plasma level of DAO and D-LA,or the expression of TNF-α and NF-κB in small intestine (P＞0.05).(3)Both exercise training for six weeks and hypoxia exposure significantly reduced the number and height of microvilli in small intestine (P＜0.01)and the occludin level in small intestine,but significantly increased the plasma level of DAO and DLA (P＜0.01),the expression of TNF-α (P＜0.01,P＜0.05)and NF-κB (P＜0.01).Meanwhile there was significant interaction between six-week exercise training and hypoxia exposure on decreasing the number (P＜0.01)and the height (P＜0.05)of microvilli in small intestine.Conclusion (1)Both intensive training and hypoxia exposure can impair intestinal mucosal barrier function and the extent of damage is correlated with the duration of training and hypoxia exposure.(2)Hypoxic exposure and intensive training may reduce the expression of occludin mRNA through increasing the expression of TNF-α and NF-κB in the small intestine,which in turn increases intestinal permeability and intestinal mucosa inju

Objective To compare the diagnosic performance of conventional 2D and 3D Cube MR imaging to identify complete and partial tears of anterior cruciate ligament(ACL).Methods Farty patients suspected of having tears of anterior cruciate ligament were prospectively studied by 3.0 T conventional 2D and 3D Cube MR imaging and arthroscopy.MR images were interpreted in consensus by two experienced radiologists,and the ACL was diagnosed as being normal,partially torn,or completely torn.Diagnostic accuracy of 2D and 3D Cube MR for the detection of both complete and partial tears of ACL was calculated using arthroscopy as the standard of reference.Area under curve (AUC) of both methods were calculated using ROC curves and were compared using Hanley& McNeil curve comparisions.Results Sixteen patients had intact ACL,12 had complete tear,and 12 had partial tear of the ACL at arthroscopy.For complete ACL tear,AUC of 2D MR and 3D Cube was 0.839 and 0.923 respectively,and there was no significant difference on ROC curves (Z =1.245,P =0.213).For partial ACL tears,AUC of 2D and 3D Cube MR were 0.643 and 0.881 respectively,and there was significant difference on ROC curves (Z =2.384,P =0.017).Conclusions Both 2D MR and 3D Cube MR have high sensitivity and specificity for identifying the complete ruptures of ACL.3D Cube MRI appears to be superior inidentifying partial rupture of ACL.

Objective To study whether solasodine hydrochloride (SBHL) could enhance the effect of arsenic trioxide in inducing apoptosis and affecting telomerase activity in cervical cancer HeLa cells. Methods Using cell culture methods, cervical cancer HeLa cells were cultured in vitro. The optimal concentration of SBHL was determined by MTT method from 0, 10, 20, 40, 80, 160, to 320 μmol/L. HeLa cells were grown in improved RPMI1640 supplemented respectively with arsenic trioxide(5 μmol/L As2O3), As2O3(5 μmol/L)+ SBHL( 40 μmol/L) and none (control group). The growth morphology of HeLa cells was observed under phase contrast microscopy after culture for 24, 48, and 72 h. Apoptosis of HeLa cells was determined under transmission electronic microscopy. The method of MTT was used to study the cell survival percentage. The technique of flow cytometry was used to measure cell cycle and cell apoptosis percentage. The method of tartrate-resistant acid phosphatase-enzyme linked immunosorbent assay (TRAP-ELISA) was used to determine telomerase activity of HeLa cells. Results Under phase contrast microscopy, in control group HeLa cells were round, densely packed; in As2O3 group the numbers of the cells were less, cell spacing increased; in As2O3 + SBHL group the cells shrinked significantly, nuclear fragmented as a petal-like, gap became larger. Under transmission electronic microscopy, there were rich microvillus on the cell surface in control group, cell intervals clear, immature connections, and the intervals did not close. The structure of the mitochondria in the cytoplasm was integrated. Most of the chromatin in the nucleus were, euchromatin and characteristics of apoptosis with heterochromatin increased and the chromatin condensed into masses, on the boundary of nuclear membrane. The microvillud on the cell surface were ruptured and decreased in As2O3 + SBHL group. The chromatin condensed into masses. The formation of apoptotic bodies was observed. The difference was statistically significant between groups in cell survival percentage at 24, 48, 72h(x2 = 10.39 , 13.88 , 17.21,respectively, all P < 0.05). Cell survival percentage in SBHL + As2O3 group (52.80%) was significantly less than that of As2O3 group(77.51%, x2 = 9.29, P < 0.05) at 72 h. In cell cycles, the difference was statistically significant between groups in C1 phase and S phase(F = 7.46,22.14, all P < 0.05), respectively. Compared with , control group[ (41.57 ± 1.56)%, (50.45 ± 2.37)%], cell percentages in S phase in As2O3 + SBHL group[(20.06 ± 4.98)%] and As2O3 group[(27.10 ± 5.32)%] were decreased(P< 0.05 or < 0.01), while cell percentage in C1 phase was increased[(58.70 ± 5.18)%, (69.67 ± 4.17)%, P< 0.05 or < 0.01]. The difference was statistically significant between groups in apoptotic percentage of HeLa cells (F = 4.01, P < 0.05). Compared with control group[ (1.18 ± 1.40)%], apoptosis percentage was significantly increased in As2O3 + SBHL group and As2O3 group [(21.08± 1.22)%, (6.04±2.53)%, P< 0.05 or < 0.01], respectively, and As2O3 + SBHL group was higher than As2O3 group(P < 0.01). The difference was statistically significant between groups in telomerase activity (F = 21.28, P< 0.05). Telomerase activity was inhibited in As2O3 group(1.214 ± 0.621) and As2O3A + SBHL group(0.865 ± 0.284) compared to control group (2.107 ± 0.057, all P < 0.05), and telomerase activity in As2O3 + SBHL group was lower than that of As2O3 group (P < 0.05). Conclusions SBHL enhances the effect of As2O3 in inducing apoptosis in HeLa cells, which is related to its inhibiting telomerase activity in HeLa cells.

BACKGROUND: Clinical observation demonstrates that accelerated fracture healing or lower limb heterotopic ossifications always occur in patients with paraplegia. It indicates that peripheral nervous system may play an important role in fracture healing process.OBJECTIVE: To observe bone histomorphometery parameter, callus formation and biochemical change during the process of fracture healing of unilateral lower limb denervated tibia.DESIGN: Self-control animal experiment.SETTING: Tianjin Hospital.MATERIALS: Totally 36 six-month-old healthy male Wistar rats, with mean body mass of 210 g, were used in this experiment.METHODS: This experiment was carried out at Animal Experimental Center of Tianjin Hospital from March 2001 to March 2004. Denervated tibia fracture model and innervated tibia fracture model were made in the same rat. Animals were executed under anaesthetic status at week 2 and week 4 after fracture. Bilateral tibias were chosen to take radiografts.Biomachamical strength was measured and non-decalcification sections were prepared to perform bone histomorphometery observation.MAIN OUTCOME MEASURES: ① Comparison of wet weight of bilateral tibias and callus of rats between two groups after fracture. ②X-ray plain film scoring. ③ Biomechanical testing of tibial samples. ④ Histomorphological observation of fracture healing RESULTS: ① Wet weight of bilateral tibia and callus of rats in denervated group was much higher than that in innervated group at weeks 2 and 4 after fracture [(0.94±0.15) vs (0.76±0.14) g, (1.06±0.26)vs (0.81±0.10) g,P ＜ 0.05]. ②In X-ray plain film scoring, callus formation was significantly increased in denervated group (P ＜ 0.01). ③In biomechanical testing of three-point bending of tibial sample, callus intensity was significantly lower at weeks 2 and 4 after fracture in denervated group than in innervated group[ (9.88±8.49)vs ( 16.62±13.38 ) N, ( 12.77±7.55 )vs (20.19±10.60) N,P ＜ 0.05]. ④Bone histomorphometery showed that compared with innervated group, mineralized bone trabecula width of denervated group was significantly reduced (P ＜ 0.05), osteoid width was increased , osteoclast index and bone absorption area were significantly increased (P ＜ 0.05), and there were no significant difference of fibroblast index and bone formation area between two groups; Compared with innervated group, mineralized deposition rate in the denervated group was significantly reduced (P ＜ 0.05), the mature time of osteoid was elongated (P ＜ 0.05).CONCLUSION: Peripheral nervous system may play an important role during early and middle period of fracture healing. Intact innervation is essential for normal fracture healing.

To observe the influence of Mycobacterium tuberculosis 19-kDa lipoprotein (Mtb P19) on function and phenotype of macrophage,the Mtb P19 was prepared from cultured Mtb H37Ra and the phorbol myristate acetate-differentiated THP-1 cells were incubated with P19 at the concentration of 10 μg/mL with 5% CO_2 at 37℃ for up to 48 hours.Supernatants were collected for TNF-α and IL-6 detection by ELISA,then the phenotype fluorescent antibodies were stained to analyze HLA-DR expression changes between control group and experimental group.Flow cytometry and microscopy was used to assay the phagocytosis in macrophages stimulated by Mtb P19.IL-6 and TNF-α in collected supernatants were detected.Results indicated that both were found significant increases and their phagocytosis were enhanced.Comparing to the control group,the mean fluorescence intensity showed a significant increase 24hs after stimulation.It presents that Mtb P19 could be able to induce macrophages activation,and it would be significantly important for protection during infection period.

ObjectiveTo improve the visualization of anteromedial and posterolateral bundles of anterior cruciate ligament (ACL) and investigate the optimal MRI plane for the bundles at a 3.0 T MR scanner.MethodsMR images of 149 knee joints were reviewed retrospectively.Display rates of AMB,PLB and their different parts (the top portion,the middle portion and the low portion) on MR different planes including axial,sagittal and coronal planes were analyzed and their differences were compared with the x2 section method.ResultsThere was no statistical difference in the display rates of two bundles of ACL between axial plane ( 115/149,77.2％ ) and coronal plane (103/149,69.1％ ) (x2 =2.4606,P ＞0.0125 ).Statistical differences were found between axial and sagittal plane,coronal plane and sagittal plane (21/149,14.1％ ) ( x2 =119.5138,92.8695 respectively,P ＜0.0125 ).There was a statistical difference for the top portion of ACL between axial plane ( 104/149,69.8％ ) and coronal plane,sagittal (0/149,0)and coronal planes ( 7/149,4.7％ ) ( x2 =135.081,159.7526 respectively,P ＜ 0.0125 ),between sagittal and coronal planes (x2 =7.1684,P ＜ 0.0125 ).For the middle portion of ACL,there was no statistical difference between axial plane ( 108/149,72.5％ ) and coronal plane (94/149,63.1％ ) (x2 =3.0120,P ＞ 0.0125 ),while statistical differences were found between axial and sagittal plane,coronal planes and sagittal plane ( 10/149,6.7％ ) ( x2 =134.7454,104.2173 respectively,P ＜ 0.0125 ).For the low portion of ACL,there was no statistical difference between axial plane ( 103/149,69.1％ ) and coronal plane (101/149,73.8％ ) (x2 =0.8065,P ＞0.0125),while statistical differences were detected between axial and sagittal plane,coronal planes and sagittal plane ( 18/149,12.1％ ) ( x2 =100.5300,115.9132,P ＜ 0.0125 ). The different parts of ACL displayed low intensity on different MR planes and normal morphology.ConclusionsACL can be displayed on conventional MR planes at a 3.0 T MR scanner to some extent.Axial and coronal planes might be the optimal MRI planes for ACL and its two bundles.

An HPLC-DAD-MS/MS method was developed for rapid analysis and identification of degradation products of buagafuran. Buagafuran and degradation products were separated on a Zorbax C8 column (5 microm, 4.6 mm x 150 mm) using acetonitrile-water (78 : 22) as mobile phase. The elutes were detected with diode array detector and tandem mass spectrometer via electrospray ionization source in positive ion mode. According to analysis of the retention time, UV spectra and MS, MS/MS data, combined with the possible degradation reaction of buagafuran, the structures of main degradation products were inferred. The results showed that six main degradation products were oxidation or peroxidation productions of buagafuran. Degradation product A was a double bond epoxidation product of buagafuran, degradation products B, C, D and E were the further oxidation products of degradation product A, degradation product F was a peroxidation product of buagafuran. The results indicated that the established method was effective in the rapid identification of the degradation products of buagafuran.

To observe the effect of Mycobacterium tuberculosis 19 kDa lipoprotein (Mtb P19) upon the expression and distribution of Toll-like receptor-2 (TLR-2) on the surface of macrophages, Mtb P19 was prepared from the cultured M.tuberculosis H37 Ra strain , and phorbol myristatye acetate (PMA)-differentiated THP-1 cells were co-cultivated with Mtb P19 at concentration of 10 g/mL and at 37 ℃ temperature and a condition containing 5% CO_2.for 6 hours.. The distribution of TLP-2 on the surface of macrophages was investigated by immuno-cellular chemical method (ICC) while the effect of Mtb P19 upon the expression of TLR on the surface of macrophages was assayed by fluorescent antibody staining. In addition,the changes of TLR-2 expression before and after the effect of Mtb P19 were investigated by flow cytometry analysis and change of TLR-2 arrangement after stimulation with Mth P19 was determined by co-focal microscopy. It was found that the TLR-2 molecules were evenly distributed on the surface of macrophages as demonstrated by ICC. The mean fluorescent intensity increased significantly after stimulation with Mtb P19 for 6 hours in comparison with that of the control group, and the patchy surface with fluorescent staining positive zones could be detected on the surface of macrophages. Nevertheless , the distribution of TLR-2 molecule in the control group appeared to be randomly dynamic. It is evident that the Mth P19 not only can induce the surface expression of TLR-2 molecules, but also cause a functional aggregation of this receptor.

Objective:To observe the expression and correlation of eIF4E and c-myc in laryngeal squamous cell carcinoma.Methods:Western blot was applied to examine the expression of eIF4E and c-myc on tumor core,transition core and tumor-free core from 36 laryngeal carcinoma patients.The results were statistically analyzed.Results:The overexpression of eIF4E and c-myc protein levels suggests an increasing tendency from tumor-free core,transition core and tumor core;and shows an significant correlation of eIF4E and c-myc expression.Conclusion:The overexpression of eIF4E and c-myc cause the laryngeal carcinoma cell malignant transformation;the correlation of eIF4E and c-myc may provide a basis for gene therapy of laryngeal carcinoma.