Objective To evaluate the performance of the Sysmex XE-5000 analyzer for analyzing atypical lymphocytes ,baso-philic granulocytes and their abnormalities warnings .Methods A total of 197 specimens with both atypical lymphocytes and baso-philic granulocytes warnings and 914 specimens with single warning of atypical lymphocytes indicated by Sysmex-5000 blood cell analyzer were collected and inspected by microscope simultaneously .Results Using microscopic examination as a standard ,baso-philic granulocytes within the normal range ,the coincidence rate of samples with both atypical lymphocytes and basophilic granulo-cytes warnings was 64 .9% ,while the coincidence rate of samples with single warning of atypical lymphocytes was 72 .5% .The for-mer was significantly lower than the latter(P<0 .05) .Conclusion When Sysmex XE-5000 indicates atypical lymphocytes and baso-philic granulocytes simultaneously ,there is interference between each other .It should be combined with microscopic examination in order to reduce the probability of missed diagnosis and misdiagnosis .

Objective To investigate the protective effect of hydrogen-rich water (HRW) on radiation-induced hematopoietic stem and progenitor cells (HSPCs) injury.Methods Totally 32 C57BL/6 mice were randomly divided into four groups with 8 mice in each group,including control,HRW,radiation and radiation + HRW.Mice in HRW and radiation + HRW groups received 0.5 ml hydrogen-rich water per day by intragastric administration 5 min before irradiation until 7 d post-irradiation.Mice in other groups received 0.5 ml distilled water.Mice in radiation and radiation + HRW group were irradiated with 2 Gy of total body irradiation.Bone marrow cells were isolated at 15 d post-irradiation,and LSK cells were examined for the percentage of hematopoietic stem and progenitor cells,the ability of colony formation and reconstitution,reactive oxygen species (ROS) levels and cell apoptosis.Results Compared with radiation group,the percentages of hematopoietic progenitor cells and LSK cells,colony number of bone marrow cells were significantly increased in radiation + HRW group (t =-4.935,-7.898,5.488,P ＜ 0.05).An elevation of donor chimerism was also found in recipient mice administered HRW after competitive bone marrow transplantation (t =-12.769,P ＜ 0.05).Compared with radiation group,the ROS levels and cell apoptosis in LSK cells were significantly decreased (t =4.380,3.954,P ＜ 0.05).Conclusions Hydrogen-rich water exhibited a protective effect on radiation-induced HSPCs injury.

Objective To investigate the protective effect of theaflavins on thymus injury caused by total body irradiation (TBI).Methods Twenty-five C57BL/6 mice were randomly divided into 5 groups:control group,4 Gy TBI group,4 Gy TBI + 25 mg/kg theaflavins group,4 Gy TBI + 50 mg/kg theaflavins group and 4 Gy TBI + 100 mg/kg theaflavins group.Thymus index and total number of thymocytes were detected at the 14th d post-irradiation to determine the optimal dose of theaflavins.According to this optimal dose,32 C57BL/6 mice were randomly divided into 4 groups:control group,theaflavins group,4 Gy TBI group and 4 Gy TBI + theaflavins group.Thymus histomorphology,CD4CD8 T cell subsets,and reactive oxygen species (ROS) in thymocytes were examined at the 14th d post-irradiation.Results The irradiated thymus exhibited decreased thymus index and total number of thymocytes (P ＜ 0.05),aberrant histomorphology and T cell subsets (P ＜ 0.05),and increased ROS level in thymocytes (P ＜ 0.05).Compared with 4 Gy TBI group,the thymus index and total number of thymocytes were significantly increased in 4 Gy TBI + 50 mg/kg theaflavins group (P ＜ 0.05).The total number of thymocytes was significantly higher in 4 Gy TBI + 50 mg/kg theaflavins group than that in 4 Gy TBI + 25 mg/kg theaflavins group (P ＜ 0.05).Therefore,50 mg/kg theaflavins was chosen as the optimal dose for subsequent experiments.Moreover,the aberrant histomorphology of irradiated thymus was alleviated by theaflavins.A decline in the percentage of CD4-CD8-T cells and an elevation of CD4+CD8-and CD4+CD8+ T cells were found in irradiated mice administered with theaflavins (P ＜ 0.05).Compared with 4 Gy TBI group,the ROS level was significantly decreased in 4 Gy TBI + theaflavins group (P ＜ 0.05).Conclusion Theaflavins exhibits a protective effect on radiation-induced thymus injury.

Objective To observe the protective effect of anthocyanin on irradiation induced bone marrow c-kit positive cell injury, and further explore its possible mechanism. Methods Mouse bone marrow c-kit positive cells were collected by cell sorting method. There were 2 groups: control group and anthocyanin group, which were sub-divided into three groups and received 0 Gy, 1 Gy and 4 Gy irradiation respectively. The control group was added 700μL cell suspension and an equal volume of serum-free hematopoietic stem/progenitor cell culture medium. The 2 × 10-5 mol/L anthocyanin was co-cultured with mouse bone marrow c-kit positive cells of anthocyanin group half an hour before irradiation exposure, then cells were cultured for 18 hours under the conventional culture conditions (37℃,5%CO2). Mouse c-kit positive cell viability was measured by bioluminescence, and which was reflected by relative light units (RLU). The ability of colony-forming units was reflected by CFU-GM. The reactive oxygen species (ROS) level and mean fluorescence intensity (MFI) ofγ-H2AX were detected by flow cytometry. Results Compared to un-irradiated control group, the cell viability and the number of CFU-GM were decreased significantly, while the ROS level and MFI ofγ-H2AX were increased in c-kit positive cells irradiated with 1 Gy and 4 Gy (P<0.05). Compared to 1 Gy and 4 Gy irradiation groups, c-kit positive cell viability and the number of CFU-GM were increased, the ROS level and MFI of γ-H2AX were decreased in anthocyanin group (P < 0.05). Conclusion Anthocyanin exhibits a promising protective effect on radiation-induced bone marrow c-kit positive cell injury, which may be related to the alleviating ROS and DNA damage in bone marrow cells.

Objective To investigate the effect of DNA-dependent protein kinase catalytic subunit (DNA-PKcs) on autophagy induction by ionizing radiation ( IR ) .Methods The cell model of knocking-down DNA-PKcs expression was constructed by transfecting HeLa cells with a pSicoR-based lentivirus vector expressing DNA-PKcs specific shRNA .Cellular growth activity and radiosensitivity were detected by cell countingkit ( CCK)-8 assay.The expression of autophagy related proteins was detected by Western blotting hybridization .Autophagy was also detected by monitoring the autophagic marker green fluorescere protein ( GFP )-light chain 3 ( LC3 ) puncta per cell under an immunofluorescent microscope .Results A cellular model of knocking-down DNA-PKcs expression was successfully generated by transfecting the specific shRNA against DNA-PKcs.Depression of DNA-PKcs significantly decreased the growth activity of HeLa cells and increased the cellular sensitivity to ionizing radiation .Both the expression changes of P 62 and LC3 proteins and immunofluorescent GFP-LC3 puncta observation indicated that knocking-down DNA-PKcs prompted the induction of autophagy by ionizing radiation . Moreover, inactivation of DNA-PKcs led to a decreased phosphorylation of mammalian target of sirolimus ( Rapamycin, RAPA) ( mTOR) at S2481 site.Conclusion Depression of DNA-PKcs expression prompts the induction of autophagy by IR and cellular radiosensitivity .mTOR signaling may be involved in the regulation of autophagy processing by DNA-PKcs.

BACKGROUND: Tendon injury is common in clinic, which is mainly treated by surgical anastomosis. Postoperative tendon healing is usually assessed through surgeons' experience due to high cost and application restrictions of MRI examination. Thus there is still a lack of a convenient and objective imaging support. With the advancement and widespread application of high-frequency ultrasound, the diagnosis rate of tendon injury has been improved remarkably; thereafter, high-frequency ultrasound used for assessing tendon injury and repair has become an issuehas become an issue of concern.OBJECTIVE: To clarify the ultrasonic imaging features of tendon repair through high-frequency ultrasound scancombined with histological examination.METHODS: This was a single-central, preoperative and self-controlled animal experiment and finished in the Central People'sHospital of Siping, China. 130 adult male Highbrow chickens were selected and were then randomized into 13 groups (n=10per group). One side of each chicken hind foot was randomly selected as experimental limb to undergo achillotomy followedby repair using the modified Kessler method (groups 2-13) or no treatment (group 1); the contralateral limb served as control.Moreover, passive flexion-extension functional training targeting the experimental limbs was performed in the groups 8-13beginning at the 1st day after surgical anastomosis, several times a day. The high-frequency ultrasound and hematoxylineosinstaining were conducted before and after chillotomy (group 1), and at 3 (groups 2 and 8), 7 (groups 3 and 9), 14 (groups4 and 10), 21 (groups 5 and 11), 35 (groups 6 and 12) and 42 (groups 7 and 13) days after surgical anastomosis, respectively.RESULTS AND CONCLUSION: The primary measurement outcomes were the repair and healing of the injured tendonas assessed by high-frequency ultrasound; the secondary outcomes were the pathological manifestations of the injuredtendon detected by hematoxylin-eosin staining. Our findings will provide preclinical proof for high-frequency ultrasounduse in the assessment of tendon injury, repair and healing as well as for the rehabilitation therapy that promotes functionrecovery in the future.

Objective To evaluate the therapeutic effect of lentinan combined with conventional antituberculous therapy in women with pelvic tuberculosis.Methods A total of 90 women with pelvic tuberculosis were enrolled and recruited into a control group and a treatment group by random number table,45 in each group.The control group was treated with anti-tuberculosis program (2HRZE/4HRE),while the treatment group was treated with lentinan tablets on the basis of the control group.Both groups were treated for 6 months.Serum levels of carbohydrate antigen 125 (CA125) and erythrocyte sedimentation rates (ESRs) were determined before and after treatment,and the therapeutic effect was evaluated.Results After treatment,the serum CA125 level (28.61 ± 9.08 U/ml vs.39.72 ± 12.13 U/ml;t=4.919,P＜0.01) and the ESR (36.13 ± 8.33 mm/h vs.41.35 ± 12.45 mm/h;t=2.338,P＜0.05) in the treatment group were significantly lower than those in the control group.The negative rate of serum CA125 after treatment than before treatment in the treatment group (93.3％ vs.20.0％;X2=46.335,P＜0.01) and the control group (82.2％ vs.8.9％;X2=37.396,P＜0.01);but there was no difference in negative rate of serum CA125 after treatment between two groups (X2=1.6571,P=0.198).The total effective rate in the treatment group was significantly higher than that in the control group (93.3％ vs.77.8％;X2=4.406,P=0.036).Conclusion Lentinan combined with conventional antituberculous therapy is effective in treatment of pelvic tuberculosis in women.

Objective To investigate the changes of the levels of interleukin(IL)-17 in bronchoalveolar lavage fluid (BALF) of BALB/c mice infected by mycoplasma pneumonia (MP).Methods Sixtyfour BALB/c mice were randomly divided into two groups,normal group (n =32) and MP infected group (n =32).The BALF and lung tissue of two groups were collected after MP infection on day 3,7,14,21,each time 8.HE staining was used to observe the pathological changes of the lung tissue.IL-17 content in BALF was detected by ELISA.Results The inflammatory changes of the lung tissue was the most obvious at 7 day,and had begun to subside at 14 day,disappeared at 21 day after MP infection.IL-17 levels in BALF were (53.783 ±2.218) pg/ml(3 day),(65.913 ± 10.693) pg/ml(7 day),(59.915 ± 8.085) pg/ml(14 day),(57.043 ± 11.997) pg/ml(21 day) in MP infection group,and (46.220 ± 3.260) pg/ml in normal group.BALF IL-17 content increased significantly in MP infection group compared with normal group(P ＜ 0.05).IL-17 content reached its peak on day 7 after MP infection (P ＜ 0.05).Conclusion MP infection increased the level of IL-17 in the mice of BALF,reaching the peak on day 7.MP infection may be involved in airway inflammation in asthma through the IL-17 pathway.

Objective To isolate and identify exosomes from serum samples of the patients with polymyositis / dermatomyositis (PM/ DM),and analyze their protein composition preliminarily.Methods Exosomes from serum samples of the patients with PM/DM were isolated and purified by the ExoQuickTM kit.The morphological characteristics and particle size of exosomes were determined by transmission electron microscope (TEM) and NanoSight analyzer,respectively.The surface markers of exosomes such as CD9,CD81 and Flotillin-2 were identified by western blot.The concentration and composition of exosome protein were determined by the BCA method and SDS-PAGE,respectively.Results The exosomes from serum samples of PM/DM patients displayed round or oval vesicles with membrane structure under TEM,and their diameter range was about (92 ± 67) nm.western blot showed that these exosomes expressed CD9,CD81 and Flotillin-2.The total protein concentrations of exosomes in the patients with PM/DM and healthy controls were 14.68 (6.00,32.55) μg/μL and 14.09 (8.00,23.28) μg/μL,respectively.SDS-PAGE showed that high-abundance proteins enriched in 55-70 kD in both PM/DM patients and healthy controls,and that there were different bands in 40-55 kD between them.Conclusion Exosomes are isolated from serum samples of the patients with PM/DM successfully,and their protein concentration and composition are analyzed preliminarily,which provides the experimental evidences for further finding differential proteins.

BACKGROUND: In the patients with congenital scoliosis, the spinal motor units exhibit developmental disorders and poor range of motion. It has been found that the compensation ability of coronal lumbosacral region (L4-S1) is associated with the occurrence of non-compensable trunk migration postoperatively.OBJECTIVE: To establish the three-dimensional finite element models of coronal lumbosacral region of normal and patients with congenital scoliosis and to compare the strain, displacement, stress and stiffness under different loading conditions among models.METHODS: One normal subject and two congenital scoliosis patients with different coronal lumbosacral region flexibility were selected, DICOM image datawere obtained by spiral CT scanning at the lumbosacral region, and then imported into MIMICS software, and a three-dimensional model was established according to the gray values of each tissue on CT,followed by simplified by GEOMAGIC, and finally imported into ABAQUS foftware to conduct a mechanic analysis under different loading conditions.RESULTS AND CONCLUSION: (1) Under different lateral forces, in the three models, the maximum stress mainly distributed on the frontal region of L4 cortical bone, and maximum displacement concentrated on L5. (2) There was no significant change in the stress distribution in the two scoliosis models, but the compensable model showed larger displacement change, and its stiffness was significantly less than that of the non-compensable model, indicating that the compensable model is easy to deform. (3) These findings suggest that three-dimensional finite element model is helpful to perform a biomechanical analysis for coronal lumbosacral region of congenital scoliosis, among which, a compensable model exhibits large displacement, suggesting a good flexibility.