Objective To observe the expression changes of microRNA-124(miRNA-124) following traumatic brain injury (TBI) in mice and investigate the correlation of miRNA-124 with neural axon regeneration.Methods Ninety-one C57BL/6 mice were assigned into TBI group (n =63) and control group (n =28) according to the random number table.Mice in TBI group were subjected to controlled cortical impact and euthanized at 12 hours and 1,3,7,14,21,28 days postinjury for the collection of brain tissue in the trauma zone.Mice in control group underwent craniectomy only.Trauma zone observation was done using the HE staining.Expression of miRNA-124 was detected using the real-time PCR.Levels of Nrp-1,Gap-43 and Tau were detected using the Western blot and immunohistochemical staining.Results After injtury,study of mice behavior and HE staining indicated the establishment of experimental model was successful.Expression of miRNA-124 reached the peak at 3 days postinjury (3.80 ± 0.22),expression of Nrp-1 reached the peak at 7 days postinjury (2.006 ±0.179),expression of Tau reached the peak at 14 days postinjury (2.063 ±0.172),and expression of Gap-43 sustained high level since 12 hours after injury(1.355 ± 0.093) (P ＜ 0.05).Count of axon marker positive cells in TBI group was the lowest at 1 day postinjury due to the direct damage and edema,and then slowly recovered.There was no significant difference in the count of axon marker positive cells between the two groups at 14,21 and 28 days postinjury (P ＞ 0.05),but the morphology in TBI group changed obviously.Although the positive cells of axon marker decreased at 1 day postinjury,expressions of miRNA-124,Nrp-1,Tau and Gap-43 in TBI group were significantly increased compared to the detections in control group (P ＜ 0.05).Conclusion Increased expression of miRNA-124 in trauma zone may closely related to axon regeneration after TBI in mice.

Objective Determining the characteristics of human fetal hepatocytes in vitro. Methods Isolating and culturing human fetal hepatocytes by stepwise trypsinization of liver fragment in vitro; collecting the culture medium to determine the secretion of AFP, ALB and the functional enzymes (including ALT, AST, GGT, ALP and LDH) of different generations in culture; determining the expression of cytochrome C by immunohistochemistry; testing the effects of sodium byturate on human fetal hepatocytes. Results Human fetal hepatocytes were polygonal epithelial cells in DMEM medium. They could be maintained for 5.5 months (about 30 passages) in vitro. They secreted ALB and functional enzymes all over their cultivation. Conclusion Human fetal hepatocytes can be maintained keeping function in vitro for several months.

Objective To identify the species of cultured cells and to detect inter-species cross contamination by polymerase chain reaction (PCR).Methods From references and NCBI database,we outsourced 32 pairs of species-specific primers targeted to genomic DNA of ten common species of cultured cells.Then we screened for optional primers with high specificity and high sensitivity.PCR amplification with these primers,the genomic DNAs isolated from the tested cell line and agarose gel electrophoresis of the PCR products were done.Mixed DNA of 10 species was used as positive-control template,and water as negative-control template.Results Ten pairs of species-specific and highly sensitive primers were selected.By PCR amplification with these primers and agarose gel electrophoresis,we may easily identify the origin of cell lines and find whether the tested cell lines are contaminated by cells of other species.Conclusion This PCR assay provides a simple,rapid,sensitive,and cost-effective method to identify cell species and detect interspecies cross-contamination.

Objective To investigate the expression levels of interferon-inducible genes (IFIT1,IFIT4,OAS1,OASL,ISG15) in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus(SLE).and the relations between these genes expression levels and disease activity are explored.Methods Sybr green dye based real-time quantitative PCR method was used to detect the expression levels (indicated as-△△Ct value) of WIT1,IFIT4.OAS1,OASL and ISG15 in 76 patients with SJJE and 54 controls.Their expression levels were compared with erythroeyte sedimentation rate (ESR),serum C reactive protein (CRP),complement C3,C4.antinuclear antibody (ANA).anti-double stranded DNA antibody.The associations between the expression levels of IFIT1,IFIT4,OASI.OASL,ISG15,ESR,CRP,complement C3,C4,ANA,anti-double stranded DNA antibody and SLEDAI scores in patients with SLE were analyzed.Results ① The expression levels of WIT1,IFIT4,OAS1,OASL and ISG15 in the SLE patients were significantly higher than those of the normal controls (P＜0.01).The expression levels of IFIT1,IFIT4,OAS1,OASL and ISG15 in active SLE patients were higher than those of inactive SLE patients (P＜0.05).The real time expression levels of IFIT1,IFIT4,OAS1.OASL and ISG15 showed positive correlations with each other (r＞0.5,P＜0.05) in patients with SLE.② The expression levels of IFIT1,IFIT4,OAS1,OASL and ISG15 were positively correlated with the SLEDAI scores (r＞0.5,P＜0.05).③ There was no correlation between ESR,CRP,complement C3,C4,ANA and the expression levels of IFIT1,IFIT4,OAS1,OASL,ISG15,SLEDAI scores except anti-double stranded DNA antibody (r＞0.5.P＜0.05).Conclusion The expression levels of IFIT1,IFIT4,OAS1,OASL and ISG15 in patients with SLE are significantly higher than those of the normal controls,and positively associated with SLEDAI scores,so they are helpful in evaluating SLE disease activity and severity.IFIT1,IFIT4,OAS1,OASL and ISG15 genes may be the potential treating targets for SLE.

OBJECTIVE:To investigate the effects of dexmedetomidine pretreatment on hepatic ischemia-reperfusion injury in patients with hepatocellular carcinoma radical surgery (HCRS).METHODS:A total of 40 patients underwent elective HCRS were randomly divided into control group (20 cases) and observation group (20 cases).Before operation,observation group was given Dexmedetomidine hydrochloride injection with loading dose of 0.7 μg/kg within 10 min,maintained at 0.4 μg/(kg.h) until the end of surgery.Control group was given constant volume of 0.9％ Sodium chloride injection as observation group and same anesthetic scheme.The levels ofALT,AST,IL-1,IL-6,CRP and TNF-α,the occurrence of ADR were observed in 2 groups before surgery,4 h,24 h,72 h after surgery.RESULTS:Before surgery,there was no statistical significance in the levels ofAST,ALT,IL-1,IL-6,CRP and TNF-α between 2 groups at different time points (P＞0.05).After surgery,the levels of AST and ALT in 2 groups were significantly higher than before surgery,and the observation group was significantly lower than the control group,with statisti-cal significance (P＜0.05).The levels of AST and ALT in 2 groups were decreasing gradually as time.4,24,72 h after surgery,the levels of IL-1 and IL-6 in 2 groups,TNF-α levels 4,24 h after surgery and CRP levels 24,72 h after surgery were significantly higher than before surgery,and the observation group was significantly lower than the control group;the levels of TNF-α in 2 groups 72 h after surgery were significantly lower than before surgery,with statistical significance (P＜0.05).The levels of IL-1,IL-6 and TNF-α were decreasing gradually as time.There was no statistical significance in the incidence of ADR between 2 groups (P＞0.05).CONCLUSIONS:Dexmedeptomidine pretreatment can relieve hepatic ischemia-reperfusion injury in HCRS patients,reduce the level of inflammatory cells without increasing the occurrence of ADR.

Objective To investigate the proliferative capacity of neural stem cells (NSCs) in rat hippocampus after traumatic brain injury (TBI) and its relationship with Janus kinase 2/signaling and transcriptional activation factor 3 (JAK2/STAT3) signaling pathway activity.Methods A total of 108 SD rats were randomly divided into control group (36 rats) and TBI group (72 rats).The TBI model was constructed by PinPointTM Precision Cortical Impactor.At 1,3,7,14,21 and 28 days after injury,the brain tissues were taken for immunofluorescence staining to detect the proliferation of NSCs [5-bromodeoxyuridine (BrdU) +/stem cell key protein-2 (Sox2) +] in hippocampus,and phosphorylated JAK2 (p-JAK2) and phosphorylated STAT3 (p-STAT3) were detected by Western blot.The expression level of p-JAK2 and p-STAT3 as well as the changing trend were analyzed.On the basis of preliminary analysis of the proliferation of NSCs and the change of JAK2/STAT3 signaling pathway activity in hippocampus,another 24 SD rats were randomly divided into TBI + normal saline group and TBI +AG490 (JAK2 specific inhibitor) group,with 12 rats in each group.At 7 days after injury,the proliferation of NSCs in hippocampus was detected by immunofluorescence staining,and the expression levels of p-JAK2 and p-STAT3 were detected by Western blot,so as to further confirm the correlation between the proliferation ability of NSCs in hippocampus and JAK2/STAT3 signaling pathway.Results Compared with the control group,the number of NSCs in the hippocampus of the TBI group and the expression of p-JAK2 and p-STAT3 increased.And the most significant increase occurred at 7 days after injury [number of NSCs:31.2 ± 4.7 in the control group,111.4 ± 8.1 in the TBI group (P ＜ 0.01);p-JAK2:1.11 ± 0.09 in the control group,2.16 ± 1.01 in the TBI group (P ＜ 0.01);p-STAT3:1.05 ± 0.06 in the control group and 2.06 ± 0.09 in the TBI group (P ＜ 0.01)].The proliferation of NSCs in hippocampus of TBI group was consistent with the change of p-JAK2 and p-STAT3 expression.Seven days after injury,the expression levels of p-JAK2 and p-STAT3 and the proliferation ability of NSCs in the TBI + AG490 were significantly decreased [p-JAK2:2.18 ± 0.15 in the TBI + isotonic saline group,1.24 ±0.10 in the TBI + AG490 group (P ＜0.01);p-STAT3:2.21 ±0.12 in the TBI + isotonic saline group,1.25 ± 0.11 in the TBI + AG490 group (P ＜ 0.01);NSCs number:112.8 ± 8.6 in the TBI + isotonic saline group,75.5 ± 6.4 in the TBI + AG490 group (P ＜ 0.05)].Conclusions The proliferation of NSCs in hippocampus of rats increased after TBI,and the activity of JAK2/STAT3 signaling pathway also increased,following the same trend.JAK2 inhibitor AG490 can reduce the activity of JAK2/STAT3 signaling pathway and the proliferation of NSCs.This can provide reference for researches on TBI promoting nerve regeneration and function repair.

Objective To explore the effects of the Calgary-Cambridge Guide training for practice nurses in promoting the nurse-patient relationship .Methods Totals of 52 practice nurses in our hospital from June 2012 to December 2012 were selected and randomly divided into the experimental group ( 25 cases ) and the control group (27 cases).Communication skills training based on the Calgary-Cambridge Guide was applied in the experimental group, while the general training materials were distributed in the control group .Nurse-patient relationship was assessed with the trust scale and the Calgary-Cambridge Guide Scale .Results After the training , the scores of respect, consistency, the authority of knowledge and technology , the sense of security and the confidence in the future of the experimental group were (3.65 ±0.45),(3.68 ±0.58),(3.62 ±0.51), (3.81 ±0.45) and (3.67 ±0.64), which were significantly higher than the scores before the training (t =-2.664,-2.712,-2.435,-3.429,-3.431, respectively;P<0.05).The scores of respect, consistency, the sense of security in the experimental group were significantly higher than those of the control group after the training ( t=2.560, 2.513, 2.231, respectively;P <0.05 ).The general score and the scores of eight dimensions of Calgary-Cambridge were significantly higher than those before (P<0.05).The general score and the scores of eight dimensions of Calgary-Cambridge of the experimental group were significantly higher than those of the control group (P<0.05).Conclusions The nurse-patient communication skills training based on the Calgary-Cambridge Guide can effectively improve the communication skills of practice nurses , and promote the nurse-patient relationship .

Objective By applying the factorial design to ascertain the best temperature and time of bathing using Xiaoyin Prescription 2 in treating patients with psoriasis vulgaris .Methods According to the inclusion criteria 120 cases with psoriasis vulgaris were selected .They were randomly divided into 4 treatment groups with 30 people for each group .Group 1 ( A1 B1 ) was treated with 42℃bathing for 30 minutes;Groups 2 ( A1 B2 ) was treated with 42℃bathing for 20 minutes;Groups 3 ( A2 B1 ) was treated with 38℃bathing for 30 minutes;Groups 4 (A2B2) was treated with 38℃bathing for 20 minutes.All of the patients were investigated by the Psoriasis Area and Severity Index ( PASI ) , Dermatology Life Quality Index ( DLQI ) and Self Rating Anxiety Scale (SAS).Results The PASI score, DLQI index and SAS score of the 120 patients were (11.33 ± 5.68), (4.78 ±3.50), (24.23 ±6.72), respectively, after the treatment, and (25.51 ±10.26), (11.60 ± 5.70), (42.37 ±8.40), respectively, before the treatment, which were significantly different (t=23.105, 17.174, 40.558, respectively;P<0.01).For Group 1, the PASI score difference before and after treatment was (16.59 ±6.46) that was significantly higher than the other three groups (F=4.242,P<0.01).The score difference of DLQI and SAS were (8.27 ±5.15), (19.93 ±4.73), higher than Group 4 whose score were (5.47 ±3.25), (16.30 ±4.82).The differences were significant (P <0.05).There was no significant interaction for time and temperature and no significant main effect of time factor ( P>0.05 ) .Only the main effect of temperature was statistically significant ( F =11.380, 6.335, 9.168, respectively; P <0.01 ). Conclusions The main effect of bathing temperature to treat patients with psoriasis vulgaris is higher than the bathing time in PASI, DLQI and SAS.Taking into account the time factor and expenditure for the patients , a herbal bathing temperature of 42 ℃ for 20 minutes is more reasonable and is worthy of promoting in clinical situations .

Objective To explore the influence of exosome-derived miR-124 on the molecular expression related to axonal regeneration after mechanical damage to cortical neurons in mice,aiming to provide experimental data for intervention in neurogenesis after traumatic brain injury (TBI).Methods The plasmid loaded with miR-124 was used to transfect the HEK293 cell line.The transfection effect was identified by real time Polymerase Chain Reaction (qPCR).The exosomes were isolated from the supematant of cultured transfected HEK293 cell line by the SBI isolation kit.The isolated exosomes were identified by electron microscopy and Western blotting,and the involved miR-124 in the exosomes was identified by qPCR.After the cortical neurons were isolated from the pregnant mice (14-17-day old) and cultured for 7 days,they were divided into 4 groups:control,damage,damage + exosomes without miR-124 and damage + exosomes with miR-124.The Petri dishes were manually scratched with a 10 μL plastic stylet needle to construct a mechanical damage in vitro in the latter 3 groups.The isolated exosomes without or with miR-124 were added into the cultured medium for culture for 72 h in the latter 2 groups,respectively.The expression ofmiR-124,NRP-1,Tau and Gap-43 was measured by qPCR and Western blotting respectively.Results The exosomes containing miR-124 were successfully obtained by plasmid transfection and the SBI isolation kit.The expression levels of miR-124,NRP-1 and Gap-43 in the damage + exosomes with miR-124 group were elevated significantly greater than in the other 3 groups (P＜0.05).The expression levels ofmiR-124,NRP-1 and Gap-43 in the damage group and damage + exosomes without miR-124 group were elevated significantly greater than in control group (P＜0.05).Conclusions The exosomes may transmit miR-124 to the cortical neurons in mice after mechanical damage and increase the expression ofmiR-124,NRP-1 and Gap-43 in the cortical neurons in mice.

Objective To explore the effect of exosome derived micro RNA (miR)-124 on activation status of microglia cells in injured brain tissues at acute phase of traumatic brain injury (TBI), and further provide theoretical references for intervention of neuroinflammation after TBI. Methods (1) In vitro cultured HEK293 cells were divided into miR-124 transfected group and control group, and miR-124 plasmids or Control siRNA by plasmid were transfected into the cells of the two groups;two-three weeks after isolation of monoclonal cell lines and two weeks after continuous culture, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the miR-124 content in cells of the two groups; the exosomes were extracted from the supernatant of cells from the two groups using SBI kit, and the morphology of the exosomes was observed under electron microscope; the expression of CD63, a surface marker molecule, was detected by Western blotting; RT-PCR was used to determine the miR-124 content in the exosomes of the two groups. (2) A total of 60 healthy male rats were randomly divided into sham-operated group (n=12), TBI group (n=12) and TBI+Exo-124 group (n=24); TBI models were constructed by controlled cortical injury device, and Exo-124 (3×109 particles) was given into the TBI+Exo-124 group via tail intravenous injection and equivalent solvent was given to the sham-operated group and TBI group 24 h after TBI; 3 d after modeling, RT-PCR was used to detect the miR-124 expression in brain tissues of the injured areas of the three groups; flow cytometry (FCM) was used to detect the percentages of Iba-1+/CD32+ and Iba-1+/CD206+ microglial cells in brain tissues; enzyme-linked immunosorbent assay (ELISA) was used to detect the expressions of interleukin (IL)-1, IL-6, IL-4 and IL-10 in the brain tissues. Results (1) RT-PCR showed that the miR-124 expression in the miR-124 transfected group was statistically higher than that in the control group (P<0.05); electron microscopy showed spherical particles with diameter about 100 nm and obvious membrane structure; Western blotting showed that the expression level of CD63 in the miR-124 transfected group was significantly higher than that in the control group (P<0.05); RT-PCR showed that the miR-124 content in the miR-124 transfected group was significantly higher than that in the control group (P<0.05). (2) The miR-124 expression in injured brain tissues of TBI+Exo-124 group was statistically higher than that in TBI group and sham-operated group (P<0.05); as compared with those in the sham-operated group, the percentages of Iba-1+/CD32+ and Iba-1+/CD206+ microglial cells and the expressions of IL-1, IL-6, IL-4 and IL-10 in the brain tissues of TBI group were significantly increased (P<0.05); as compared with the TBI group, the TBI+Exo-124 group had significantly decreased percentage of Iba-1+/CD32+ microglial cells and significantly increased percentage of Iba-1+/CD206+ microglial cells, statistically decreased IL-1 and IL-6 expressions, and statistically increased IL-4 and IL-10 expressions (P<0.05). Conclusion Exosome-derived miR-124 promotes the polarization of microglia cells from M1 to M2 and reduces neuroinflammation at acute phase of TBI.