[Objective]To construct arecombinant eukaryotic expression plasmid carrying human BMP-7 gene which was transfected into bone marrow stromal cells(BMSCs) in vitro and expressed.[Method]Clone human BMP-7 cDNA from a Chinese woman placenta was recombinanted with plasmid pcDNA3.1,to construct eukaryotic expression carrier of recombinaut pcDNA3.1-hBMP-7.The BMSCs were isolated from rabbit bone marrow and cultured in vitro.They were divided into three groups: pcDNA3.1-hBMP-7 transduced group;pcDNA3.1 transduced group;untransduced group.The expression of hBMP-7 was detected by RT-PCR,Immuneohistochemistry.ALP,Collogen,Osteocalcin production were detected.[Result]The pcDNA3.1-hBMP-7 transduced BMSCs expressed hBMP-7 at both mRNA and protein levels.ALP activity was detected in pcDNA3.1-hBMP-7 transduced cells from day 2 to day 10,peaking on day 8.Collogen,osteocalcin production were elevated significantly too(P

Objective To investigate the role of phosphatidyl-inositol 3-kinase-Akt (PI3k-Akt) signal pathway in the attenuation of ischemia-reperfusion (I/R) injury by sevoflurane preconditioning in isolated rat hearts. Methods Ninety-six adult male SD rats weighing 220-280 g were randomly divided into 6 groups ( n = 16 each): sham operation group (group S); I/R group; sevoflurane preconditioning group (group SP); wortmannin group (group W); dimethyl sulfoxide (DMSO) group (group D) and sevoflurane preconditioning + wortmannin group (group SW) . Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5%C02 at 37 ℃ . The hearts were continuously perfused for 180 min in group S. After 15 min of equilibration, the isolated hearts were subjected to 30 min of ischemia followed by 120 min of reperfusion in SP, W, D and SW groups. Croups SP, W, D and SW received 10 min of perfusion with K-H solution containing 2. 4% sevoflurane, 100 nmol/L wortmannin, 20 μmol/L DMSO, and 2.4% sevoflurane + 100 nmol/L wortmannin, respectively, followed by 5 min washout before I/R. Eight hearts in each group were selected and HR, left ventricular end-diabetic pressure (LVEDP), left ventricular developed pressure (LVDP), and ± dp/dtmax were recorded at the end of equilibration and at 15 min of reperfusion, Myocardial tissues were obtained at 15 min of reperfusion for determination of apoptosis (by TUNEL) and phosphorylated Akt (p-Akt) expression (by Western blot) . Another 8 hearts were selected at 120 min of reperfusion for determination of myocardial infarct size by TTC staining. Result Compared with group S, LVDP and ± dp/dt,^ were significantly decreased and LVEDP was significantly increased in groups I/R, SP, W, D and SW, and myocardial p-Akt expression was up-regulated in groups I/R, SP and D ( P ＜ 0.05). Compared with group I/R, LVDP and ± dp/dtmax were significantly increased, LVEDP and apoptosis index were significantly decreased, myocardial p-Akt expression was up-regulated, and myocardial infarct size was significantly reduced in group SP (P ＜0.05) . Conclusion Activation of PI3K-Akt signal pathway is involved in the attenuation of I/R injury by sevoflurane reconditioning in isolated rat hearts.

ObjectiveTo investigate the protective effect of sevoflurane postconditioning (Sevo-Postcon)on the hearts isolated from rats with diabetes mellitus (DM) of different duration against ischemia-reperfusion (I/R)injury.MethodsSeventy-two pathogen-free male SD rats weighing 200-240 g were randomly divided into 3 groups ( n =24 each):group Ⅰ rats without DM (C) ; group Ⅱ rats with 2 week DM (2w DM) and group Ⅲ rats with 6 week DM (6w DM).DM was produced by intraperitoneal (IP) 1％ streptozocin (STZ) 60 mg/kg and confirmed by fasting blood glucose concentration ＞ 16.7 mmol/L in groups Ⅱ and Ⅲ.Hearts were isolated from rats and perfused with Krebs-Henseleit buffer (KHB) in a Langendorff apparatus.After a 15 min stabilization period,the isolated hearts were subjected to 30 min of global no-flow ischemia followed by 75 min of reperfusion.Twelve hearts in each group were perfused after ischemia with KHB saturated with 3％ Sevo for 15 min followed by perfusion with regular KHB for 60 min.LVEDP,LVDP, ± dp/dt and HR were measured and recorded after 15 min stabilization (T0,baseline) and at 15 and 75 min of reperfusion (T1,2 ).Myocardial specimens were obtained at 15 min of reperfusion (T1) for detection of p-Akt expression (by Western blot analysis).Infarct size was determined at 75 min of reperfusion (T2).ResultsSevo-Postcon significantly improved cardiac function,reduced infarct size and up-regulated p-Akt expression in groups Ⅰ (C) and Ⅱ (2w DM),while in group Ⅲ (6w DM) Sevo-Postcon did not cause any change in cardiac function,infarct size and p-Akt expression as compared with the isolated hearts without Sevo-Postcon.ConclusionThe cardioprotective effect of Sevo-Postcon can be attenuated with increasing duration of DM by impairing PI3K/Akt signaling pathway.

Aim To determine the function of nuclear factor-κB ( NF-κB ) in immunological liver injury of rat model and its effect on CYP2E1 expression, content and metabolic activity. Methods The immunological liver injury rat model was prepared by injection of Ba-cillus Calmette Guérin ( BCG,125 mg · kg-1 ) for 14 days. The hepatic tissue injury was revealed by hema-toxylin and eosin ( HE ) method and serum concentra-tion of alanine aminotransferase( ALT) , aspartate ami-notransferase ( AST ) respectively. CYP450 total con-tent in hepatic homogenate was determined by spectro-photography. The expression of CYP2E1 protein was detected by Western blot analysis. The enzyme kinetics of CYP2 E1 probe drug chlorzoxazone was evaluated by high-performance liquid chromatography ( HPLC ) as-say. Results The results showed that BCG-pretreat-ment ( 125 mg · kg-1 ) significantly increased the weight of liver and spleen, serum levels of ALT and AST(P<0. 01) , and decreased CYP2E1 expression, content and metabolic activity ( P <0. 05 ) . Adminis-tration of ammonium pyrrolidine dithiocarbamate (PDTC) (50, 100, 200 mg·kg-1) reversed the a-bove hepatic injury stimulated by BCG in vivo. Moreo-ver, PDTC dose-dependently inhibited the down regu-lation of CYP2 E1 ( P<0. 05 ) . Conclusion Passiva-tion of NF-κB can inhibit the down regulation of CYP2 E1 in liver tissue of immunological liver injury rats;NF-κB may be involved in CYP2 E1 down-regula-tion.

OBJECTIVE:To evaluate the efficacy and safety of new agent zaleplon in treatment of insomnia.METHODS:A randomized,double-blind,controlled and dosage-adjusted clinical trial was conducted.The subjects had been treated with zaleplon5～10mg/d and zopicolone7.5～15mg/d for14days,respectively.RESULTS:The scores of sleep dysfunction rating scale at endpoint were significantly reduced as compared with the baseline in both groups(P

BACKGROUND:Under the in vitro conditions of cell harvesting, culture, and transplantation, whether bone marrow stromal cells (BMSCs) can be effectively applied in local gene therapy remains unclear.OBJECTIVE: To construct a recombinant eukaryotic expression plasmid carrying human bone morphogenetic protein-7 (hBMP-7) gene, and to expect to enhance osteoinductive properties of rabbit BMSCs transfected.DESIGN, TIME AND SETTING: A cell-genomics in vitro observation was performed at the Laboratory of Scientific Research, Second Affiliated Hospital of Harbin Medical University between July 2006 and July 2007.MATERIALS: Human healthy fresh placental tissue was provided by the Department of Gynaecology and Obstetrics, Second Affiliated Hospital of Harbin Medical University. Written informed consent was obtained from the women. One healthy male New Zealand rabbit was provided by the Laboratory Animal Center, Harbin Medical University.METHODS: hBMP-7 gene was cloned from human placental tissue to construct a recombinant eukaryotic expression plasmid carrying hBMP-7 gene by conjugating with eukaryotic expression vector pcDNA3.1. BMSCs were isolated from rabbit bone marrow and cultured in vitro. Then they were divided into 3 groups: pcDNA3.1-hBMP-7-transfected, pcDNA3.1 -transfected, and untransfected. 5×106 BMSCs were inoculated into a 60 mm3 flask containing antibiotic-free medium 1 day prior to transfection.MAIN OUTCOME MEASURES: RT-PCR and immunohistochemistry were employed to detect hBMP-7 expression in BMSCs, alkaline phosphatase activity, hydroxypreline content, and osteocalcin production in each group. RESULTS: After 72-hour transfection, a 1.3 kb fragment was seen in the pcDNA3.1-hBMP-7-transfected group, showing brown granules in the endochylema, but not seen in the pcDNA3.14ransfected and untransfected groups. ALP activity in the pcDNA3.1-hBMP-7-transfected group significantly increased at 2 days after transfection, peeked at 8 days, and still increased at 10 days. At each time point, alkaline phosphatase activity, hydroxyproline content, and osteocalcin production were significantly higher in the pcDNA3.1-hBMP-7-transfected group than in the pcDNA3.1 -transfected and untransfected groups (P<0.05 or P<0.01).CONCLUSION: Recombinant eukaryotic expression vector pcDNA3.1- BMP-7 was constructed successfully. Results indicated that hBMP-7 was expressed in BMSCs sufficiently and was involved in inducing differentiation of BMSCs into osteoblasts. The method would provide substantial basement for hBMP-7 gene therapy.

Objective To explore the effect of peer education on the practical teaching of fundamental nursing for male nursing students .Methods Thirty-six male nursing undergraduate students from Grade 2011 were chosen as the observation group , and received the peer education .Thirty-two male nursing undergraduate students from Grade 2010 were chosen as the control group , and received the traditional teaching .The teaching effect was evaluated by the score of final examination and questionnaire in two groups .Results The scores of theory and practice test were respectively (57.68 ±4.34), (26.34 ±3.11) in the observation group, and were higher than (54.34 ±4.15), (24.58 ±2.93) in the control group, and the differences were statistically significant (t=3.233,2.393, respectively; P <0.05).The result of questionnaire on the teaching effect showed that the professional identification , learning interest , self-confidence , communications and collaboration abilities in the observation group were better than those of the control group (P<0.05), and no differences were found in the abilities of operation, problem analysis and innovation between two groups (P >0.05). Conclusions Peer education can improve the professional identification in the male nursing students , and increase their learning interest and self-confidence , and improve their scores of theory and practice test in the fundamental nursing , and increase their abilities of communications and collaboration .

Objective To investigate the role of miR-194-3p in regulating functional changes in osteoblasts in a simulated microgravity environment and to provide a theoretical foundation for understanding the mechanical response mechanisms of osteoblasts in extreme mechanical environments.Methods The effects of microgravity on osteoblasts were simulated by using a rotary cell culture system.MC3T3-E1 osteoblasts were transfected with an miR-194-3p inhibitor and changes in proliferation,differentiation,apoptosis,and mineralization were assessed using MTT assay,RT-PCR,Western blot,double fluorescence staining,and alizarin red staining.Results Elevated expression of miR-194-3p under simulated microgravity conditions led to the suppression of osteoblast proliferation,differentiation,and mineralization to a certain extent,while promoting osteoblast apoptosis.However,transfection with the miR-194-3p inhibitor significantly downregulated miR-194-3p expression and partially reversed the reduced osteoblast proliferation,decreased expression of osteogenic differentiation markers such as ALP,OCN,and COL-I genes and proteins,decreased bone mineralization nodules,and increased osteoblast apoptosis induced by microgravity exposure.These findings indicated that miR-194-3p effectively ameliorates abnormal osteoblast function under microgravity conditions.Conclusions MiR-194-3p acts as a negative regulatory factor in the mechanical responses of osteoblasts under simulated microgravity.

Objective:To study the life characteristics and related risk factors of patients with depression in Shandong Province.Methods:Based on the 2015 mental epidemiological survey database in Shandong Province,a total of 832 patients with depression,807 high-risk individuals with depression,and 819 low-risk individuals were diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders,Fourth Edition(DSM-Ⅳ)and the Structured Clinical Interview for DSM-Ⅳ-TR Axis I Disorders,Research Version(SCID-I/P).In 2020,SCID-I/P was used for re diagnosis,and the General Health Questionnaire(GHQ-12),Simple Quality of Life question-naire,Pittsburgh sleep quality index(PSQI),Childhood Trauma Questionnaire(CTQ),Social Support Rating Scale(SSRS),Global Pain Scale(GPS),Montreal Cognitive Assessment(MoCA),Simplified Coping Style Question-naire(SCSQ)were used for evaluation.Compare changes in the quality of life of depression patients and construct a risk factor model.Results:Patients with depression had lower scores on the simple quality of life questionnaire at baseline and at retest after 5 years than those in the high-and low-risk groups,those in remission of depression had higher scores on the simple quality of life questionnaire at baseline and at retest after 5 years than those in non-re-mission,and those with new-onset disorder in the high-and low-risk groups had lower scores on the simple quality of life questionnaire at baseline and at retest after 5 years than those with no-onset disorder(Ps＜0.001).Depres-sion diagnosis and PSQI scale scores at baseline negatively predicted at retest after 5 years(β=-0.06,-0.15),while coping style tendencies at baseline positively predicted(β=0.06).The simple quality of life questionnaire at baseline negatively predicted depression diagnosis at retest after 5 years,GHQ-12 scores at retest after 5 years,and PSQI scale scores at retest after 5 years(β=-0.11,-0.17,-0.09),while the simple quality of life question-naire at baseline positively predicted coping style tendencies at retest after 5 years(β=0.13).Depression diagnosis at retest after 5 years,GHQ-12 scores at retest after 5 years,PSQI scale scores at retest after 5 years,coping style tendencies at retest after 5 years,SSRS scale scores,CTQ scale scores,GPS scale scores,and the simple quality of life questionnaire at baseline all influenced the simple quality of life questionnaire at retest after 5 years through ei-ther direct or indirect pathways.Conclusion:It suggest that the quality of life is lower in patients with depression than in the general population.Depression diagnosis,sleep,mental health,pain,social support,childhood trauma and coping are direct and indirect risk factors affecting life.

Objective:To observe the differences of macular microvascular structure between recurrent and non-recurrent macular edema (ME) secondary to central retinal vein occlusion (CRVO) after intravitreal injection of ranibizumab (IVR), and to preliminarily analyze the correlation between recurrence and ME.Methods:A prospective clinical observational study. Forty-five patients (45 eyes) diagnosed as CRVO with ME were included in this study in Tianjin Medical University Eye Hospital from January 2020 to December 2021. There were 22 males (22 eyes) and 23 females (23 eyes). All cases were unilateral. The average age was 61.11±10.88 years old. All patients received IVR treatment once a month for 3 consecutive months. ME were regressive after the initial three treatments. The patients were divided into recurrent group (21 cases, 21 eyes) and non-recurrent group (24 cases, 24 eyes) based on ME recurrence at 6 months after ME resolution. All patients underwent best corrected visual acuity (BCVA), intraocular pressure, and optical coherence tomography angiography (OCTA). OCTA was used to scan the macula in the area of 3 mm×3 mm, and the vessel density (VD) of superficial capillary plexus (SCP), deep capillary plexus (DCP), fovea and parafovea before and after treatment was measured. Foveal retinal thickness, foveal avascular zone (FAZ) area, perimeter of FAZ (PERIM), avascular index of FAZ (AI), VD within 300 μm width of FAZ range (FD-300). Foveal VD included superficial and deep retinal VD (SFVD, DFVD); parafoveal VD included superficial and deep retinal VD (SPFVD, DPFVD). Taking the initial three treatments as the observation time point, the changes of the parameters of the two groups were compared. Comparison between the recurrent and non-recurrent group was performed by two independent sample t-tests. Receiver operating characteristic (ROC) curve analysis was used to measure the area under the curve (AUC) of VD for predicting the recurrence of ME. Results:There were no significant differences in age ( t=1.350), IOP ( t=1.929), SFVD ( t=-1.716), DFVD ( t=-1.143), CRT ( t=-1.207) and AI ( t=1.387) between the recurrent and non-recurrent group ( P>0.05). There were significant differences in times of anti-VEGF therapy ( t=5.912), BCVA ( t=5.003), SVD ( t=-4.617), SPFVD ( t=-4.110), DVD ( t=-5.503), DPFVD ( t=-4.772), FAZ area ( t=2.172), PERIM ( t=2.606) and FD-300 ( t=-3.501) between the recurrent and non-recurrent group ( P<0.05). ROC curve analysis showed that the AUC of DVD in predicting the recurrence of ME was highest, with 0.921, and the threshold was 37.65%. The sensitivity and specificity were 91.7% and 85.7%, respectively. Conclusions:The SVD, SPFVD, DVD, DPFVD and FD-300 in the recurrence group are significantly lower than those in the non-recurrence group, while the FAZ area and PERIM are significantly higher than those in the non-recurrence group. DVD≤37.65% can be used as the best threshold for predicting the recurrence of ME.