Objective To establish the rapid PCR am plification program and system and to verify the technical indexes. Methods PCR m ultiplex and capillary electrophoresis detection of 24 autosom al STR loci and one Y-STR loci using the 6-color fluorescence m arking technology, as w ell as Amelogenin and Y-InDel. Meanw hile, sensitivity, specificity, identity, stability, m ixing and a batch of sam ple tests w ere investigated, and the genotype of various routine sam ples and degraded, exfoliated cell sam ples w ere observed. Results The sensitivity of the system w as 0.062 5 ng. In addition, the genotype could be detected accu-rately only around 65 m in via rapid am plification. The species-specificity w as high and the genotyping of all kinds of dry blood specim ens of filter paper and m ixed, degraded, exfoliated cell sam ples w ere accu-rate. Conclusion The rapid am plification system can significantly im prove the detection rate, and obtain accurate and stable genotyping results, w hich m ay be im portant im plications for the establishm ent of STR database and study on population genetics and forensic identification.

Objective To observe the extracellular content of dopamine (DA)and expression of D1 receptors in hippocampal dentage gyrus (DG)in the model rats with vascular dementia (VD),and to investigate the relationship between them.Methods 12 male SD rats were randomly divided VD group and sham-operation group,and the VD model was prepared by permanent bilateral carotid occlusion.The extracellular content of DA in the DG was determined by in vivo microdialysis and HPLC,and the expression of D1 receptors was measured by immunehisto-chemistry.Results The DA content in the DG of the rats in VD group was lower than that in sham-operation group (P <0.05).The number of D1 receptor-positive cells in the DG hilus in VD group was increased compared with sham-operation group (P < 0.05),whereas the expression of D1 receptor in DG granule cell layer did not change (P > 0.05).Conclusion The DA content in the hippocampal DG is decreased in the VD rats,and its function may be compensated by the up-regulation of D1 receptors in the DG hilus.

Objective To investigate whether the sodium arsenite inhibiting proliferation of hepatocellar carcinoma(HCC) cells and having a relation with the expression of promyelocytic leukemia (PML) protein .Methods The immunohistochemistry , Western blot analysis ,immunofluorescence assay and quantitative PCR were used to examine the PML gene and protein expression in HCC tissue and cells .Results The immunohis to chemical staining showed that the PML protein was expressed in nucleus of well‐differentiated ,moderately differentiated and poorly differentiated HCC tissues randomly selected from 15 cases of HCC under‐going hepatic resection .Western blot analysis showed that PML protein was expressed at varying levels in all 24 HCC tissue sam‐ples ,HuH7 ,HepG2 ,Hep3B ,SMMC‐7721 and L02 cells .The immunofluorescence assay demonstrated that PML protein grains were found in the nucleis of the HuH7 ,HepG2 ,Hep3B and SMCC‐7721 cells ,in which HuH7 and Hep3B expressed more PML proteins than HepG2 and SMMC‐7721 cells .24 HCC tissue samples ,Hep3B ,HepG2 ,SMCC‐7721 and HuH7 cells all expressed PML gene .Sodium arsenite not only down‐regulated the PML protein expressed in HuH7 and primary HCC cells ,but also inhibited the HCC cell growth of HepG2 ,Hep3B ,HuH7 and SMMC‐7721 ,with the exposure time extension ,the inhibiting effect of sodium arsenite on HCC cells was enhanced .Conclusion Both HCC cell lines and tissues generally express ther PML protein and gene , PML protein may serve as the molecular basis for the direct targeting effects of arsenical agents .

BACKGROUND: Osteogenic ability of bone morphogenetic protein-2 has been wel documented in many experiments, but a series of factors are involved in osteogenesis induction that is a complex network adjustment process. OBJECTIVE: To quantitatively determine the level of insulin-like growth factor I during the lumbar spinal fusion of rabbits induced by recombinant human bone morphogenetic protein-2. METHODS: Sixty adult male New Zealand white rabbits were randomly divided into three groups: bone autograft, bone al ograft or composite bone (bone al ograft with 75 μg recombinant human bone morphogenetic protein-2) was implanted into the L5-6 intertransverse process of rabbits, respectively. At days 7, 14, 21, 28, 35 after implantation, formed cal us was taken to detect the expression of insulin-like growth factor I using real-time fluorescence quantitative PCR. RESULTS AND CONCLUSION: In the three groups, the expression of insulin-like growth factor I gradual y increased with implantation time, peaked at 28 days and then decreased. At 7 days after implantation, the expression of insulin-like growth factor I was higher in the autograft group than the composite and al ograft groups (P < 0.05); at 14 days, the expression of insulin-like growth factor I was higher in the autograft and composite groups than the al ograft group (P < 0.05); at 21, 28 and 35 days, the expression of insulin-like growth factor I was higher in the composite group than the autograft and al ograft groups (P < 0.05). These findings indicate that recombinant human bone morphogenetic protein-2 can improve the expression of insulin-like growth factor I effectively during the lumbar spinal fusion.

BACKGROUND:Knee osteoarthritis is a chronic disease characterized as cartilage degeneration and hyperostosis to impact the soft tissues around the joints, thereby resulting in joint pain, and it can be aggravated and lead to joint deformity after weight-bearing. OBJECTIVE:On the basis of imaging parameters of knee osteoarthritis patients and normal controls, a model was established. And then, knee squatting movement was simulated on the model and stress data from the cartilage surface of patients and normal controls were colected and compared, thereby to obtain the stress characteristics of the cartilage surface in knee osteoarthritis patients. METHODS: There were 30 knee osteoarthritis patients and 30 healthy volunteers in the study. CT and MRI data from these participants were colected and analyzed using Mimics software and simpleware software to establish a model. Based on this model, cartilage-related stress data were harvested from knee osteoarthritis patients and healthy volunteers and compared to summarize the stress characteristics of the articular cartilage in patients with knee osteoarthritis. RESULTS AND CONCLUSION:In knee osteoarthritis patients, the stress on the cartilage surface from standing to squatting was increased in a nonlinear manner. During the process of squatting, the mediolateral stress was higher in the knee osteoarthritis patients than healthy controls (P < 0.05); however, there was no difference in the proximodistal and anteroposterior cartilage surface stress between the two groups (P > 0.05). These findings indicate that the knee osteoarthritis musculoskeletal model and finite element model are established successfuly, and these models are closer to the real motion characteristics of the knee joints, based on which, quantitative biomechanical data of the knee can be provided for knee osteoarthritis treatment.

Objective To observe the effects of the extract from daylily on the learning and memory in depression model rats, and to explore its possible mechanism. Methods Rats were randomly divided into control group, model group ( CUMS), the positive group (CUMS + fluoxetine) , high, medium and low dose group (stress + daylily extract in different doses), 12 rats in each group. The depression model was established by combining separation and chronic unpredictable stress. Body mass, open-field-test and sugar consumption experiment were used to evaluate the changes of behaviors in rats. And morris water Maze test was used to evaluate the ability of learning and memory. Results There was no statistically distinction between the rats of each group on weight and the behavioral indicator before modeling. Compared with the control group, the vertical movement,horizontal movement of open box test in the model group were reduced (P＜0.05 ) ,and sugar consumption and preference degree decreased (P<0.05 ).The target quadrant time, platform resident time, effective area residence time and crossing platform times in the water maze test of the model group was less than those of the control group. The daylily effect was evaluated at 0, 7, 14, 21, 28, 35, 42 days respectively post treatment. There were significant differences in depression behaviors between model group and daylily group(P＜0.05). And each indicator in the water maze test of the daylily group (high, medium dose) was more than that of model group (P ＜ 0. 05). Control group, model group, positive group, high, medium and low dose group, vertical movement scores were (41.83 ± 17.63; 8.14 ±4.23; 12.73 ±7.21; 23.17 ± 18.75; 8.38 ±3.46; 13.50 ±5.44); horizontal movement scores were (69.92 ±34.04; 28.33 ±20.36; 62.25 ± 15.72; 69.42 ±35.17 ; 49.08 ±32.85; 48.08 ±21.19). Conclusion Daylily may be partially work on the depression of rats, with some antidepressant effect, meantime,daylily can improve the ability of learning and memorizing of the depressed rats.

Objective To observe the ultrastructure changes of hippocampal dentage gyrus (DG) and spatial learning and memory ability changes in rats models of vascular dementia (VD), and investigate the relationship between them.Methods Twelve adult male SD rats were randomly divided into sham-operated group and VD model group (n=6);the VD rat models were prepared by improved permanent bilateral carotid occlusion.The spatial learning and memory abilities of rats were assessed by Morris water maze (MWM), and the ultrastructures of DG were detected by transmission electron microscope.Results (1) In the place navigation trial of MWM, the mean escape latency in VD group was significantly longer than that in sham-operated group (P＜0.05);in the spatial probe trial, the number of platform crossings in VD group was markedly smaller as compared with that in sham-operated group (P＜0.05).(2) The changes in ultrastructures of hippocampal DG in VD rats were as follows: the number of synaptic vesicles was reduced;the rough endoplasmic reticulum was reduced and arranged in disorder,and free ribosome was increased;the membrane and cristae of mitochondria in the synaptosoma were blurred, and the cristae was fractured;the synaptic cleft was blurred.Conclusion The spatial learning and memory disabilities in VD rats may be associated with injury of ultrastructures in hippocampal DG.

Objective To explore the effect of quality control circle ( QCC ) on non-standard implementation of aseptic operation related medical staffs. Methods Through QCC activities, a total of 950 operation related medical staffs had been investigated non-standard implementation status of aseptic operation, and analyzed the causes, made out and carried out its countermeasures. Results Non-standard implementation of aseptic operation declined from 690 person-time to 175 person-time, and the target completion rate was 105%, improvement rate 75%, of which spreading towels did not reached standard declined from 327 person-time to 99 person-time. The off operation table people, who was too near from the sterile stage, declined from 134 person-time to 33 person-time, while not using the non-contact wear gloves declined from 71 person-time to 12 person-time. Conclusions QCC activities can significantly reduce the person-time of non-standard implementation of aseptic operation and effectively improve the quality of work.

Objective:To study the effects of umbilical cord mesenchymal stem cells (UCMSCs) loaded by graphene oxide (GO) on cartilage repair in two KOA animal models.Methods:30 male New Zealand rabbits aged 12 weeks were randomly divided into A group ( n=15) and B group ( n=15). In the A group, the KOA model was established by the improved Hulth and cartilage defect method, and in the B group, the KOA model was established by the modified papain controlled-release injection method. After the modeling, the rabbits model in each group were divided into 4 subgroups, including blank control group ( n=3), GO group ( n=4), UCMSCs group ( n=4) and GO+UCMSCs group ( n=4). In these subgroups, the rabbit models were respectively treated injected with 0.5 ml of NaCl solution with 9 g/L, GO granular lubricant (GO with 30 μg/ml and solvent with hyaluronic acid with 0.25%), UCMSCs suspension (5×10 6 /ml), and mixed suspension of UCMSCs loaded by GO (GO with 30 μg/ml and UCMSCs with 5×10 6/ml) in right knee joint cavity. The serum levels of NO, collagen type Ⅱ(COL-Ⅱ), glycosaminoglycan (GAG), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). Results:Eight weeks after the treatment, the serum levels of NO, IL-6 and TNF-α in GO+UCMSCs group were lower than those of the blank control group (all P<0.01), and the serum levels of COL-Ⅱ and GAG in GO+UCMSCs group were higher than those of the blank control group (all P<0.01). The serum NO level of the blank control group in group A was lower than that of the blank control group in group B [(22.097±0.352) ng/ml vs (23.662±0.056) ng/ml, P<0.05]. The serum COL-Ⅱ levels of the UCMSCs group and GO+UCMSCs group in group A were higher than those of group B respectively [(15.589±0.063) ng/ml vs (14.429±0.092) ng/ml, and (19.372±0.063) ng/ml vs (16.257±0.416) ng/ml, all P<0.01]. The serum GAG levels of the blank control group and the GO+UCMSCs group in group A were higher than those in group B respectively [(23.832±0.891) ng/ml vs (18.709±0.552) ng/ml, and (37.439±2.155) ng/ml vs (26.554±0.450) ng/ml, all P<0.01). The serum IL-6 levels of the blank control group and the GO+UCMSCs group in group A were lower than those in group B respectively [(16.082±0.323) ng/ml vs (18.367±0.861) ng/ml, P<0.05; (7.426±0.294) ng/ml vs (8.680±0.242) ng/ml, P<0.01]. The serum TNF-α levels of the blank control group and the GO+UCMSCs group in group A were lower than those in group B respectively [(9.466±0.177) ng/ml vs (10.013±0.197) ng/ml, P<0.05; (5.139±0.183) ng/ml vs (6.210±0.058) ng/ml, P<0.01]. Conclusions:GO loaded UCMSCs can promote the secretion of chondrocytes in rabbit KOA models, reduce inflammatory levels in joints, and play a role in cartilage repair.

Objective:To study the clinical efficacy and lower limb temperature difference of bipolar radiofrequency combined with ozone in the treatment of lumbar disc herniation(LDH).Methods:A retrospective study was conducted on 105 inpatients with LDH. All patients were treated with bipolar radiofrequency combined with ozone. The interdisc impedance of the main electrode was measured before and after treatment, and the absolute temperature difference of infrared thermal imaging of both lower extremities before and after treatment was analyzed. Vasual analogue scale (VAS) score, JOA score and modified Macnab standard were used to evaluate the curative effect after treatment.Results:The VAS score after treatment was significantly lower than that before treatment( P<0.05), and the JOA score after treatment was significantly higher than that before treatment( P<0.05). The intervertebral disc impedance after treatment was significantly lower than that before treatment( P<0.05). The temperature difference between leg and plantar after treatment was significantly lower than that before treatment (all P<0.05). Conclusions:Bipolar radiofrequency combined with ozone therapy can relieve pain and improve function in patients with LDH. Infrared thermal imaging can sensitively reflect the changes of body surface temperature of lower extremities in patients with LDH, and has a certain value in the diagnosis and curative effect evaluation of LDH.