MicroRNA-22-3p Regulates the Expression of Kruppel-like Factor 6 to Affect the Cardiomyocyte-like Differentiation of Bone Marrow Mesenchymal Stem Cell / 中国医学科学院学报

Objective To explore the effect of microRNA-22-3p (miR-22-3p) regulating the expression of Kruppel-like factor 6 (KLF6) on the cardiomyocyte-like differentiation of bone marrow mesenchymal stem cell (BMSC). Methods Rat BMSC was isolated and cultured,and the third-generation BMSC was divided into a control group,a 5-azacytidine(5-AZA)group,a mimics-NC group,a miR-22-3p mimics group,a miR-22-3p mimics+pcDNA group,and a miR-22-3p mimics+pcDNA-KLF6 group.Real-time fluorescent quantitative PCR (qRT-PCR) was carried out to determine the expression of miR-22-3p and KLF6 in cells.Immunofluorescence staining was employed to detect the expression of Desmin,cardiac troponin T (cTnT),and connexin 43 (Cx43).Western blotting was employed to determine the protein levels of cTnT,Cx43,Desmin,and KLF6,and flow cytometry to detect the apoptosis of BMSC.The targeting relationship between miR-22-3p and KLF6 was analyzed by dual luciferase reporter gene assay. Results Compared with the control group,5-AZA up-regulated the expression of miR-22-3p (q=7.971,P<0.001),Desmin (q=7.876,P<0.001),cTnT (q=10.272,P<0.001),and Cx43 (q=6.256,P<0.001),increased the apoptosis rate of BMSC (q=12.708,P<0.001),and down-regulated the mRNA (q=20.850,P<0.001) and protein (q=11.080,P<0.001) levels of KLF6.Compared with the 5-AZA group and the mimics-NC group,miR-22-3p mimics up-regulated the expression of miR-22-3p (q=3.591,P<0.001;q=11.650,P<0.001),Desmin (q=5.975,P<0.001;q=13.579,P<0.001),cTnT (q=7.133,P<0.001;q=17.548,P<0.001),and Cx43 (q=4.571,P=0.037;q=11.068,P<0.001),and down-regulated the mRNA (q=7.384,P<0.001;q=28.234,P<0.001) and protein (q=4.594,P=0.036;q=15.945,P<0.001) levels of KLF6.The apoptosis rate of miR-22-3p mimics group was lower than that of 5-AZA group (q=8.216,P<0.001).Compared with the miR-22-3p mimics+pcDNA group,miR-22-3p mimics+pcDNA-KLF6 up-regulated the mRNA(q=23.891,P<0.001) and protein(q=13.378,P<0.001)levels of KLF6,down-regulated the expression of Desmin (q=9.505,P<0.001),cTnT (q=10.985,P<0.001),and Cx43 (q=8.301,P<0.001),and increased the apoptosis rate (q=4.713,P=0.029).The dual luciferase reporter gene experiment demonstrated that KLF6 was a potential target gene of miR-22-3p. Conclusion MiR-22-3p promotes cardiomyocyte-like differentiation of BMSC by inhibiting the expression of KLF6.

Effect of Puerarin Combined with Tanshinone ⅡA on t-PA, PAI-1 and Oxidative Stress in DM Rats with Vascular Lesions / 中国实验方剂学杂志

Objective:To observe the protective effect and mechanism of combination of puerarin combined with tanshinone ⅡA on diabetes mellitus (DM) rats with vascular lesions. Method:The SD rats (fed with high-fat diet) were administrated with streptozotocin(STZ) through intravenous injection to make the model of diabetic vascular lesions. The successfully modeled rats were randomly divided into the model control group, the high-dose group (0.5 g·kg-1+1.0 g·kg-1), the middle-dose group (0.25 g·kg-1+0.5 g·kg-1), the low-dose group (0.05 g·kg-1+0.1 g·kg-1), the puerarin group (0.25 g·kg-1), the tanshinone ⅡA group (0.5 g·kg-1) and the positive control group (Metformin, 0.09 g·kg-1). Each group was administrated with drugs respectively by gavage for 70 days. After intervention in each group, the general conditions and body weight of the rats were observed. The contents of blood grucose and blood lipids were determined by automatic biochemical analyzer. The contents of insulin, advanced glycation end products (AGEs), superoxide dismutase(SOD), glutathione peroxidase (GSH-Px) in serum, the contents of tissue plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1) and thromboxane B2 (TXB2) in plasma, as well as the contents of AGEs and oxidized low-density lipoprotein(ox-LDL) in aorta homogenate were detected by enzyme-linked immunosorbent assay(ELISA). The content of malondialdehyde(MDA) in serum was determined by chemical colorimetry. Pathological changes of coronary tissue were observed by htoxylin eosin(HE) staining. The expression of PAI-1 protein of aorta was observed by immunohistochemistry. Result:Compared with the normal control group, in the model group, the levels of blood grucose and blood lipids (PPPP2 in plasma (PPPPPPPPP2 in plasma (PPPPPPPConclusion:Puerarin combined with Tanshinone ⅡA could relieve vascular lesions of DM rats. The mechanisms may be related to the reduction of oxidative stress and the regulation of coagulation-fibrinolysis system.

Analysis of urinary storage symptoms following transurethral resection of the prostate / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the lower urinary tract symptoms (LUTS), especially those in the urinary storage phase, following transurethral resection of the prostate (TURP), and to improve the postoperative management and patients' quality of life after TURP.</p><p><b>METHODS</b>A total of 86 patients with benign prostate hyperplasia (BPH) underwent TURP, and were interviewed on urinary symptoms at 1, 3, 7, 15 and 30 days after removal of the catheter. The patients were divided into two groups according to whether they had preoperative detrusor instability and/or compliance of the bladder (Group A) or not (Group B), and observed for the changes in IPSS scores and urinary storage symptoms after removal of the catheter.</p><p><b>RESULTS</b>Complete follow-ups were achieved in 71 cases, 28 with detrusor instability and/or compliance of the bladder and the other 43 without. Their IPSS scores on the 1st, 3rd, 7th, 15th and 30th day after removal of the catheter were 8.1 +/- 2.5, 7.2 +/- 3.1, 6.3 +/- 3.8, 5.3 +/- 4.2 and 2.4 +/- 3.4, respectively, with statistically significant differences between the 7th and the 1st as well as the 30th and the 15th day (P < 0.05), but not between the 1st and the3rd nor the 15th and the 7th day (P > 0.05). On the 1st day, the cardinal symptoms in the urinary storage phase were urinary frequency, urgency and incontinence; the scores on IPSS and urinary storage symptoms were 10.4 +/- 3.3 and 9.3 +/- 3.8 in Group A and 6.2 +/- 2.8 and 5.2 +/- 2.7 in Group B, with significant differences between the two groups (P < 0.05). After treatment with tolterodine and alpha-adrenoreceptor inhibitor, neither IPSS scores nor the scores on urinary storage symptoms showed any significant differences between Groups A and B on the 15th and 30th day (P > 0.05).</p><p><b>CONCLUSION</b>The lower urinary tract symptoms following TURP, especially those in the urinary storage phase, are correlated with preoperative bladder function, and getting improved gradually after surgery.</p>

Clinical effect of electroacupuncture combined with psychologic interference on patient with Internet addiction disorder / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe, adopting randomized controlled method, the therapeutic effect of electroacupuncture (EA) combined with psychologic interference in patients with internet addiction disorder (IAD).</p><p><b>METHODS</b>Forty-seven patients with IAD were assigned to two groups treated respectively with psychotherapy alone (A, 23 cases) and EA plus psychotherapy (B, 24 cases). The psychotherapy was conducted by cognition and behavior method, once every 4 days, for 10 times totally. EA was applied at acupoints Baihui, Sishencong, Hegu, Taichong, Neiguan, Sanyinjiao, etc. once every other day, for 20 times. Changes of scoring by IAD self-rating scale (ISS), anxiety self-rating scale (SAS), self-rating depressive scale (SDS), Hamilton depression scale (HAMD), Hamilton anxiety scale (HAMA) and self-rating sub-health scale (SRSHS) before and after treatment were observed.</p><p><b>RESULTS</b>The total effective rate was 91.3% (21/23) in Group B, better than that (59.1%, 13/22) in Group A. By the end of this study, all scores in Group B, except HAMD, were significantly lower than those in Group A respectively, i.e., for IAD, 33.20 +/- 4.53 vs. 44.00 +/- 5.81; for SAS, 30.90 +/- 6.30 vs. 39.60 +/- 5.80; for SDS, 35.38 +/- 4.59 vs. 39.60 +/- 6.33; for HAMA, 7.50 +/- 2.54 vs. 12.70 +/- 3.68; for SRSHS, 39.60 +/- 5.66 vs. 48.40 +/- 6.91, showing statistical significances (P < 0.05).</p><p><b>CONCLUSION</b>Using psychologic interference alone or combined with EA can significantly reduce the ISS score and significantly reduce anxiety and improve self-conscious health status in patients with IAD, but the effect obtained by the combined therapy is better.</p>

Effects of electroacupuncture combined with psychologic interference on anxiety state and serum NE content in the patient of internet addiction disorder / 中国针灸

<p><b>OBJECTIVE</b>To observe therapeutic effect of electroacupuncture (EA) on internet addiction disorder (lAD) and to preliminarily probe the mechanism.</p><p><b>METHODS</b>Forty-seven cases of TAD were randomly divided into a psychotherapy group and an EA plus psychotherapy group. The psychotherapy group were treated with cognition and behavior therapy, once every 4 days, 10 sessions constituting one course; the EA plus psychotherapy group were treated with EA at Baihui (GV 20), Sishencong (EX-HN 1), Hegu (LI 4), Taichong (LR 3), Neiguan (PC 6), Sanyinjiao (SP 6), etc. once every other day, for 20 sessions, in combination with the same psychotherapy as that in the psychotherapy group. Changes of score of lAD, score of anxiety self-rating scale (SAS), score of Hamilton anxiety scale (HAMA) and serum norepinephrine (NE) content before and after treatment were observed.</p><p><b>RESULTS</b>The total effective rate was 91.3% in the EA plus psychotherapy group and 59.1% in the psychotherapy group, the former being better than the latter (P<0.05). The scores for IAD, SAS, HAMA, and the se-rum NE content after treatment were significantly decreased in the two groups (both P<0.01). Those in the EA plus psychotherapy group were significantly lower than those in the psychotherapy group (P<0.05).</p><p><b>CONCLUSION</b>Electroacupuncture combined with psychologic interference can significantly improve anxiety state and the mechanism is possibly related with the decrease of NE in the body.</p>

Establishment of the methods to detect nucleophosmin gene and its mutation in acute myelogenous leukemia / 中国实验血液学杂志

This study was aimed to establish the PCR methods to detect nucleophosmin (NPM) gene and its mutation. 2 leukemia cell lines and 23 specimens from patients with acute myelogenous leukemia (AML) were investigated. The level of NPM mRNA was detected by RT-PCR. The exon-12 of NPM gene in leukemia cell lines was amplified by PCR and sequenced. Using the plasmid containing cDNA of NPM mutation A as a positive template, the PCR procedure to detect mutation A was established and evaluated. Then, the mutation of NPM was analyzed in 23 AML specimens. The results indicated that the expression level of NPM in leukemia cell lines was higher than that in normal cells. Different overexpression levels of NPM mRNA were found in all 23 AML specimens. PCR indicated that mutation had been not occurred at NPM exon-12 in THP1 and K562 cells, but a T base was deleted at 3' untranslated region of NPM gene in K562 cells. The PCR used for directly detecting NPM mutation A can specially amplify the NPM mutation gene. The method was reproducible, whose coefficient of variability was 1.6% and 3.1% in intra-and inter-assays respectively. The lowest detectable limit was 100 pg cDNA. Using the PCR methods, NPM mutation A could be detected in 2 out of 23 AML specimens, but NPM mutation A was not found in THP1 and K562 cells. It is concluded that the RT-PCT method detecting NPM mRNA level and the PCR method detecting directly NPM mutation are established. NPM mRNA is overexpressed in leukemia cells; NPM mutation A occurs in some AML patients.

Effect of suyu capsule on behavior and injury of hippocampal neurons in depression model mice / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of Suyu capsule on behavior, injury of hippocampal neurons and Ca2+ ion in hippocampal synaptic in the depression model mice.</p><p><b>METHOD</b>Sixty male Kunming mice were randomly divided into 5 groups, the control group, the model group and three Suyu capsule groups (the doses were 22.8, 11.4, 5.7 g x kg(-1) respectively). The model was established by separation and chronic unpredictable mild stimulation. The increased weight and crossing score, rearing score were measured by open-field and sweet water consumption of mice. Cone cell and configuration of neuron in CA1, CA3 region of hippocampus were observed by Nissl. The concentration of hippocampal synaptic Ca2+ ion was detected by fluorimetry.</p><p><b>RESULT</b>Comparing with the mice of control, the increased weight was slowered ( P < 0.01), the scores of rearing and crossing were decreased (P < 0.01), sweet water consumption were decreased too (P < 0.01), numbers of cone cell in CA3 region of hippocampus were decreased obviously (P < 0.01), and Ca2+ ion in hippocampal synaptic was increased obviously. Comparing with the mice of model, Suyu capsule (22.8 g kg(-1)) could increase the increased weight on the 14th and 21 st day obviously (P < 0.05); Suyu capsule (22.8 g x kg(-1)) could increase the scores of crossing obviously (P < 0.05), Suyu capsule (22.8, 11.4 g x kg(-1)) could increase the scores of rearing obviously (P < 0.01, P < 0.05); Suyu capsule (22.8, 11.4, 5.8 g x kg(-1)) could increase sweet water consumption obviously (P < 0.01, P < 0.05, P < 0.05; Suyu capsule (22.8, 11.4, 5.8 g x kg(-1)) could increase numbers of cone cell in CA3 region of hippocampus obviously (P < 0.01, P < 0.05, P < 0.05); Suyu capsule (22.8, 11.4, 5.8 g x kg(-1)) decreased Ca2+ ion in hippocampal synaptic with dose-effect relationship (P < 0.01, P < 0.01, P < 0.05).</p><p><b>CONCLUSION</b>Suyu capsule can improve all the symptoms of the depression model mice and protect injury of hippocampal neurons in the depression model mice. The possible mechanism of action is to restrict Ca2+ ion overfreight.</p>

Effect of Angelica on Proliferation of Neural Stem Cells in Dentate Gyrus of Juvenile Rats after Intrauterin Hypoxia / 实用儿科临床杂志

Objective To explore the effect of intrauterin hypoxia on proliferation of neural stem cells(NSCs)in dentate gyrus of juve-nile rats and the interventive effects on NSCs.Methods Thirty-nine adult female SD rats bred at laboratory.At 14 days of pregnancy,conceiving female SD rats were randomly divided into 3 groups:control group(n=12),hypoxia group(n=12)and angelica group(n=15);the conceiving rats were put into 3 gases incubator(130 mL/L O2,temperature 25 ℃,relative humidity 70%-75%,CO2 0.3-0.4 mL/L),hypoxia times(2 h/d),hypoxia days(pregnant the 14th d,15th d,16th d)to make intrauterin hypoxia model.In hypoxia group,1 hour before hypoxia,the rats were injected saline from caudal vein,saline for 7 days;In angelica group,the rats were injected angelica injection,angelica for 7 days,other steps were same.In control group,the rats were injected saline,saline for 7 days,no hypoxia.Producing naturally,after born 40 days,took juvenile rats' brains from every group randomly:control group(n=12),hypoxia group(n=12)and angelica group(n=15),then immobilization,paraffin imbedding,slicing,common immunohistochemistry staining were used to detect the Nestin positive substances.Nestin positive substances expression were observed under the microscope(?200).Statistical analysis was used.Results The average of positive staining area(Area)of Nestin of hypoxic group were larger than that of control group,it had significance(P0.05),the average of density and IOD of 3 groups had no significances.Conclusion Hypoxia environment(130 mL/L)can promote NSCs of juvenile rats to proliferate,angelica injection can reduce the NSCs to proliferate after intrauterin hypoxia.

Effect of Intrauterin Hypoxia on Proliferation of Neural Stem Cells and Memory and Learning Capacity of Juvenile Rats / 实用儿科临床杂志

Objective To explore the effect of intrauterin hypoxia on proliferation of neural stem cells(NSCs)and memory and learning capacity of juvenile rats.Methods Conceiving female SD rats(n=20)were divided randomly into 2 groups:control group(n=10)and hypoxia group(n=10).The rats of hypoxia group were put into 3 gases incubator to make the injured brain newborn rats model,rats of control group were not done so.The juvenile rats were taken to do the experiment of water maze after they were born in 30 d,then killed when the experiment was over,the brains of the rats were taken,immobilization,paraffin imbedding,slicing(hippocampus),common immunohistochemistry staining were used to detect the Nestin positive NSCs.Results Optical density of Nestin positive cells in hippocampus dentation of hypoxia group(0.16?0.10)was higher than that of control group(0.15?0.09)(t=3.24 P