Objective: To investigate the effect and possible mechanism of berberine (Ber) on myocardial injury induced by exhaustion exercise (Ee). Methods: Forty healthy male SPF Sprague-Dawley rats were randomly divided into 5 groups using the random unit group design method: control group, Ee group and Ee plus Ber group (low: 50 mg·kg^-1·d^-1, medium: 100 mg·kg^-1·d^-1 and high dose: 150 mg·kg^-1·d^-1, n=8 each). Ber (1.5 ml) or equal volume saline was given per gavage for 14 days. Rats assigned to Ee groups underwent Ee swimming once daily and rats in control group remain sedentary. After 14 days, echocardiographic measurements were performed and left ventricular ejection fraction (LVEF) and fractional shortening (LVFS), left ventricular diastolic diameter (LVIDd) and left ventricular systolic diameter (LVIDs) were obtained. The morphological structure of heart was detected by HE and Masson staining. Serum superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by enzyme-linked immunosorbent assay. Cardiomyocytes apoptosis was detected by TUNEL method. The protein expression of myocardial hypertrophy marker protein B-type natriuretic peptide (BNP) and apoptotic marker protein (Bcl-2, Bax) in rat myocardial tissue was detected by Western blot. Results: (1) Both LVFS and LVEF were significantly lower, and LVIDs and LVIDd were significantly larger in Ee group than those in control group (all P<0.01). The LVFS and LVEF in medium dose of Ber and high-dose Ber groups were significantly higher, and the LVIDs and LVIDd were significantly smaller than those in Ee group (all P<0.01). (2) The results of HE staining showed that the myocardial cells in control group were closely arranged, regular, normal in morphology, clear in structure, and uniform in staining. The myocardial cells of rats in Ee group were disarranged, cell staining was uneven, and vacuoles appeared in the cytoplasm. The disorder of myocardial cell arrangement and unequal staining in the medium dose of Ber were attenuated than in Ee group. The Masson staining results showed that the myocardial cells in control group were closely arranged, regular, normal in shape, clear in structure, and rarely blue-stained (fibrosis). Myocardial cells in rats in Ee group showed obvious fibrosis. The myocardial cell fibrosis in rats with medium dose of Ber was significantly reduced than exercise group. (3) MDA content in myocardial tissue of rats in Ee group was significantly higher than that of control group, and MDA content in myocardial tissue of rats in medium dose of Ber group was significantly lower than in Ee group (P<0.01). The SOD activity of myocardial tissue in rats was significantly lower than that of control group, while that of rats with medium dose of Ber was significantly higher than that of rats in Ee group (P<0.01). (4) TUNEL staining results showed that only a small amount of apoptosis myocardial cells were seen in control group, and a large number of apoptosis myocardial cells were seen in rats in Ee group. However, the number of apoptotic cardiomyocytes in medium dose of Ber was significantly lower than that in Ee group. The AI of rat cardiomyocytes was significantly higher than that of control group (P<0.01), and the AI of rat cardiomyocytes in median dose of Ber group was significantly lower than in Ee group (P<0.01). (5) BNP and Bax protein expression in the myocardial tissues of rats in Ee group were significantly higher than in control group (P<0.01). BNP and Bax protein expression in the myocardial tissues in median dose of Ber group were significantly lower than that of Ee group (P<0.01). The myocardial protein expression level of Bax was significantly higher, and the myocardial protein level of Bcl-2 was significantly lower in Ee group than in control group (both P<0.01), treatment with median dose of Ber could partly reverse above changes (both P<0.01). Conclusion Ber can attenuate exhaustion exercise induced myocardial injury and remodeling in rats, and the beneficial effects of Ber might possibly be mediated by reducing free radical release and cardiomyocytes apoptosis.

Objective To explore the protective effect and mechanism of swimming rehabilitation training on learning and memory impairment of cerebral ischemia reperfusion gerbil. Methods Forty adult healthy male gerbils were randomly divided into sham group,sham+swimming group (Sham+S group),cere-bral ischemia / reperfusion group ( I/R group), cerebral ischemia/reperfusion+swimming group ( I/R+S group),with 10 rats in each group. The gerbil models of cerebral ischemia/reperfusion in I/R group and I/R+S group were established by blocking bilateral common carotid artery,while for gerbils in Sham group and Sham+S group, only bilateral common carotid arteries of gerbils were exposed, but no arteries were clamped. Morris water maze was used to detect the changes of learning and memory function in rats. Oxida- tive stress injury in hippocampal neurons was detected by detection kit analysis. And the expression of Bax, Bcl-2 and CaMK Ⅱ protein in hippocampal tissue was detected by Western blot. Results Compared with Sham group,the gerbils in I/R group had longer positioning cruise time and less shuttle times ( both P<0. 01). Compared with I/R group,the positioning cruise time and shuttle times in I/R+S group were signifi-cantly shortened and increased respectively (both P<0. 01). Compared with sham group( SOD:(123. 13± 7. 50)U/mg,GSH:(42. 10±2. 17) μg/g,GSH-Px:(61. 37±2. 51) μg/g,MDA:( 2. 91± 0. 23) nmol/mg), the activities of SOD,GSH,GSH-Px in I/R group decreased significantly,while the content of MDA increased significantly(SOD:(75. 50±6. 96)U/mg,GSH:(22. 50±1. 64) μg/g,GSH-Px:(33. 15±2. 04)μg/g,MDA:(5. 96±0. 32)nmol/mg;all P<0. 01). Furthermore,compared with I/R group,the above indexes in I/R+S group were significantly reversed(SOD:(110. 30±5. 90)U/mg,GSH:(34. 31±1. 73)μg/g,GSH-Px:(50. 13 ±2. 31)μg/g,MDA:(3. 57±0. 29) nmol/mg;all P<0. 01). Compared with Sham group,the expression of Bax protein in hippocampus of gerbils in I/R group was increased,while the expression of Bcl-2 protein and p-CaMK Ⅱ protein was decreased (all P<0. 05). Compared with I/R group,the expression of Bax protein in hippocampus of gerbils in I/R+S group was decreased,while the expression of Bcl-2 protein and p-CaMK Ⅱprotein was increased (all P<0. 05). Conclusion Swimming rehabilitation training can improve learning and memory impairment of gerbils after ischemia-reperfusion through anti-oxidative stress and anti-apoptosis, which may be related to CaMK Ⅱ signaling system.

Objective To investigate the effect and possible mechanism of berberine (Ber) on myocardial injury induced by exhaustion exercise (Ee).Methods Forty healthy male SPF Sprague?Dawley rats were randomly divided into 5 groups using the random unit group design method: control group, Ee group and Ee plus Ber group (low: 50 mg·kg-1·d-1, medium: 100 mg·kg-1·d-1 and high dose: 150 mg·kg-1·d-1, n=8 each). Ber (1.5 ml) or equal volume saline was given per gavage for 14 days. Rats assigned to Ee groups underwent Ee swimming once daily and rats in control group remain sedentary. After 14 days, echocardiographic measurements were performed and left ventricular ejection fraction (LVEF) and fractional shortening (LVFS), left ventricular diastolic diameter (LVIDd) and left ventricular systolic diameter (LVIDs) were obtained. The morphological structure of heart was detected by HE and Masson staining. Serum superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by enzyme?linked immunosorbent assay. Cardiomyocytes apoptosis was detected by TUNEL method. The protein expression of myocardial hypertrophy marker protein B?type natriuretic peptide (BNP) and apoptotic marker protein (Bcl?2, Bax) in rat myocardial tissue was detected by Western blot. Results (1) Both LVFS and LVEF were significantly lower, and LVIDs and LVIDd were significantly larger in Ee group than those in control group (all P<0.01). The LVFS and LVEF in medium dose of Ber and high?dose Ber groups were significantly higher, and the LVIDs and LVIDd were significantly smaller than those in Ee group (all P<0.01). (2) The results of HE staining showed that the myocardial cells in control group were closely arranged, regular, normal in morphology, clear in structure, and uniform in staining. The myocardial cells of rats in Ee group were disarranged, cell staining was uneven, and vacuoles appeared in the cytoplasm. The disorder of myocardial cell arrangement and unequal staining in the medium dose of Ber were attenuated than in Ee group. The Masson staining results showed that the myocardial cells in control group were closely arranged, regular, normal in shape, clear in structure, and rarely blue?stained (fibrosis). Myocardial cells in rats in Ee group showed obvious fibrosis. The myocardial cell fibrosis in rats with medium dose of Ber was significantly reduced than exercise group. (3) MDA content in myocardial tissue of rats in Ee group was significantly higher than that of control group, and MDA content in myocardial tissue of rats in medium dose of Ber group was significantly lower than in Ee group (P<0.01). The SOD activity of myocardial tissue in rats was significantly lower than that of control group, while that of rats with medium dose of Ber was significantly higher than that of rats in Ee group (P<0.01). (4) TUNEL staining results showed that only a small amount of apoptosis myocardial cells were seen in control group, and a large number of apoptosis myocardial cells were seen in rats in Ee group. However, the number of apoptotic cardiomyocytes in medium dose of Ber was significantly lower than that in Ee group. The AI of rat cardiomyocytes was significantly higher than that of control group (P<0.01), and the AI of rat cardiomyocytes in median dose of Ber group was significantly lower than in Ee group (P<0.01). (5) BNP and Bax protein expression in the myocardial tissues of rats in Ee group were significantly higher than in control group (P<0.01). BNP and Bax protein expression in the myocardial tissues in median dose of Ber group were significantly lower than that of Ee group (P<0.01). The myocardial protein expression level of Bax was significantly higher, and the myocardial protein level of Bcl?2 was significantly lower in Ee group than in control group (both P<0.01), treatment with median dose of Ber could partly reverse above changes(both P<0.01). Conclusion Ber can attenuate exhaustion exercise induced myocardial injury and remodeling in rats, and the beneficial effects of Ber might possibly be mediated by reducing free radical release and cardiomyocytes apoptosis.

Objective To explore the effect of lncRNA of growth arrest-specific 5 (lncRNA GAS5) on the radiosensitivity of colon cancer cells by targeting miR-223.Methods The expressions of lncRNA GAS5 in a few of colon cancer cell lines were detected by real-time quantitative PCR (qPCR).The cell lines with low expression level of lncRNA GAS5 were selected for subsequent study.The effect of overexpression of lncRNA GAS5 on the radiosensitivity of colon cancer SW480 cells was detected by cell cloning experiments.The target gene miR-223 of lncRNA GAS5 was predicted and validated by the bioinformatics database starBase and dual luciferase reporter assays.qPCR was used to detect the expression of miR-223 in various colon cancer cell lines and the influence of lncRNA GAS5 overexpression on the expression of miR-223 in SW480 cells.Results Compared with normal human colonic epithelial cells (NCM460),the expressions of lncRNA GAS5 in the colon cancer SW480,LOVO,HT-29 and SW620 cell lines were significantly lower(t =15.25,8.69,14.42,11.62,P ＜ 0.05),with the lowest level in SW480 cells.Both overexpression of lncRNA GAS5 and down-regulation of miR-223 significantly increased the radiosensitivity of colon cancer cells by decreasing cell survival fraction (at 8 Gy,lncRNA GAS5,t =13.51,P ＜ 0.05;anti-miR-223,t =14.93,P ＜ 0.05)and promoting apoptosis (lncRNA GAS5,t =8.30,P ＜ 0.05;anti-miR-223,t =7.32,P ＜ 0.05).Bioinformatics analysis showed that the 3'sequence of lncRNA GAS5 contained the binding sites with miR-223.After overexpression or downregulation of lncRNA GAS5,the expression of miR-223 was enhanced or reduced.Conclusions The lncRNA GAS5 promotes the apoptosis of colon cancer cells and inhibits its survival by targeting miR-223 expression,thereby increases the radiosensitivity of colon cancer cells.

Objective To study the effect of treadmill exercise on spatial learning and memory of autism rats and its possible mechanism.Methods Ten normal wistar rats were chosen as control group,and autism-like animal models were induced in other 20 rats by intraperitoneal injection of 600 mg/kg valproate acid (VPA);the autism models were randomly divided into vehicle group and training group (n=10).The rats in the training group were forced to run on a treadmill for 30 min once a d for 28 consecutive d,while rats in vehicle group and control group were housed on static treadmill only.Then,learning and memory capabilities of all rats were tested by Morris water maze test.Superoxide dismutase (SOD) and catalase (CAT) activities and methylene dioxyamphetamine (MDA) content of the hippocampus tissues were subsequently detected by ELISA Kits.Brain-derived neurotrophic factor (BDNF),phosphorylate (p)-protein kinase (Akt),B-cell lymphoma-2 (Bcl-2),and Bcl-2 associated X protein (Bax) expressions in the hippocampus CA1 were detected by Western blotting.Nissl staining and NeuN immunohistochemistry were utilized to observe the number of normal and mature neurons.Results (1) As compared with rats in the vehicle group,the rats in the training group had significantly shortened escape latency and increased times of passing through the platform (P＜0.05).(2) Nissl staining indicated that as compared with the rats in the vehicle group,the rats in the training group had significantly smaller number of apoptotic neurons in CA1 area of the hippocampus (P＜0.05).(3) NeuN immunohistochemistry showed that number of NeuN positive cells of the vehicle control group was significantly smaller than that of the control group,while that of the training group was significantly larger than that of the vehicle group (P＜0.05).(4) Western blotting showed that MDA content significantly decreased,glutathione content significantly increased,and SOD and glutathion peroxidase activities significantly enhanced in the training group as compared with those in the vehicle group (P＜0.05);The expressions of Bcl-2,BDNF,p-Akt/Akt,p-calmodulin-dependent kinase Ⅱa (CaMKⅡ)/CaMKⅡ and pcAMP-response element binding protein(CREB)/CREB were significant up-regulated,and caspase-3 and Bax expressions were significant down-regulated in the training group as compared with those in th evehicle group (P＜0.05).Conclusion The treadmill exercise may improve the spatial learning and memory abilities of autism rats;treadmill exercise strengthens the brain antioxidant ability and plays a neuroprotective role in hippocampal neuronal cells.

Objective To construct a prostate cancer specific oncolytic adenovirus armed with a fusion protein gene , PSA-IZ-CD40L, and to evaluate its oncolytic efficiency and immune activation ability in vitro.Methods Prostate Specific Antigen (PSA) gene, CD40L-N and CD40L-C genes were obtained from cDNA of LNCaP cells and Jurkat cells using poly-merase chain reaction (PCR) or nested-PCR, respectively.PSA,Linker,CD40L-N and CD40L-C were linked sequentially to generate fusion protein gene PSA-IZ-CD40L (PL) by overlapping PCR.Then, prostate specific oncolytic adenovirus PL-carrying gene, Ad-PL-PPT-E1A,was constructed using the oncolytic adenovirus system , which was based on Adeasy sys-tem.PC3M cells were infected by Ad-PL-PPT-E1A at serial multiplicity of infection (MOI), and the apoptosis was detec-ted by flow cytometry at several time points post-infection.For immune activation detection , PC3M cells were infected with Ad-PL-PPT-E1A at a MOI of 50, and the cell lysate was collected at 48 h post-infection.Peripheral blood mononuclear cells derived (PBMCs) from healthy donors were stimulated by the lysate from PC 3M cells or Ad-PL-PPT-E1A infected PC3M cells before proliferation was assayed using cell counting kit-8 (CCK8).Results Fusion protein gene, PSA-IZ-CD40L, was successfully constructed and cloned into the prostate cancer specific adenovirus to generate Ad -PL-PPT-PL. The expression of E1A and PL protein could be detected by reverse transcription PCR and Western-blotting.Cytopathic effect was observed in PC3M cells infected with Ad-PL-PPT-E1A.Furthermore, the apoptosis rate reached 70.67% ± 2.98%at 48 h post-infection with 200 MOI Ad-PL-PPT-E1A.Compared with the lysate of PC3M cells, that from Ad-PL-PPT-E1A infected cells could promote the proliferation of PBMCs .Conclusion We have constructed a prostate cancer spe-cific oncolytic adenovirus armed can fusion protein gene PL , Ad-PL-PPT-E1A, which could kill PC3M cells effectively and enhance the proliferation of PBMCs in vitro.

ObjectiveTo investigate the husband and wife psychological and behavioral intervention on high-risk pregnant women,pregnancy outcome and negative emotions.Methodsin line with the number of highrisk pregnancy diagnostic criteria for pregnant women into the group of order packets were completed by the clinical observation of high-risk pregnant women in the intervention group (A) 31 cases,32 cases of high-risk pregnant women in the control group (group B),spouses of pregnant women in the intervention group (Group C) 31 spouses of pregnant women in the control group ( group D).Pregnant women in group A and group B underwent outpatient conventional high-risk pregnancy management,group A,group C received 16 weeks of husband and wife jointly participate in the key psychological problems,negative emotion coping skills to learn,couples communication skills,learning,family and social support operations,rehabilitation and faith strengthening and other intervention as the core content.Quality delivery of newborns,asphyxia,anxiety and depression in pregnant women and their spouses before and after intervention the overall incidence of anxiety and self-assessment scale(SAS) score,the score of the Self-Rating Depression Scale (SDS),the Family APGAR Index Questionnaire score (observation of high-risk pregnancy APGAR) and other changes.ResultsThe average body weight of newborns:the intervention group A (3.12 ± 0.69) kg,than in group B (2.29 ± 0.78) kg,a statistically significant difference ( t =2.3148,P =0.024) ;asphyxia:group A was 12.9％ and 34.4％ in group B,the difference was statistically significant (x2 =4.0018,P=0.0455) ;natural birth rate:58.1％ in group A,group B 25％,a statistically significant difference (x2 =7.1023,P=0.0077) ;the rate of cesarean section:29.0％ in group A,group B,59.4％,a statistically significant difference ( x2 =5.8713,P =0.0154 ) ; anxiety and depression in pregnant women:the total incidence after the intervention group A was 19.4％,46.9％ in Group B,the difference was significant (x2 =5.3664,P=0.0205) ;maternal spouse anxiety and depression:in the overall incidence of A group of 9.7％ after the intervention group B 31.2％,the difference was statistically significant (x2 =4.4745,P =0.0344 ) ;APGAR score:after the intervention of high-risk pregnant women in group A (9.42 ± 1.53),Group B (7.71 ± 1.56),group A better than group B,the difference was statistically significant ( t =4.3910,P =0.000),intimacy,emotional degree,the growth degree,cooperation degree,adapt to the degree of five factor scores in group A than group B,a statistically significant difference (P ＜ 0.05,P ＜ 0.01 ).ConclusionHigh-risk pregnant women and their spouses have a severe negative emotional reaction,the husband and wife psychological and behavioral intervention on the improvement of high-risk pregnant women,pregnancy outcome and negative emotions have an important role.

Objective To evaluate the prognosis influencing factors of early non-small cell luny cancer (NSCLC) after radiotherapy.Methods 81 early NSCLC patients received definitive radiotherapy and were eligible.Among these patients,60 were diagnosed as squamous cell carcinoma,16 were adenocarcinoma and 5 were diagnosed through imaging instead of pathology.45 patients received conventional radiotherapy,36 patients received three dimensional conformal radiotherapy (3D-CRT),All of them received a total dose of 50-96 Gy with a median dose of 67.8 Gy. Kaplan-Meier survival curves and Cox regression model analysis were applied to evaluate the survival and prognostic factors. Results The median survival time was 34 months.The 1-,3- and 5-year survival rates (OS) were 88.7 ％,41.9 ％,21.8 ％,respectively.Karnofsky performance status≥80,Clinical stage, diameter≤4 cm and the therapeutic effect were associated with improving overall survival.Cox hazards model showed that Karnofsky performance status≥ 80 and diameter≤4 cm were likely to be independent positive prognostic factors. Conclusion Karnofsky performance status and tumor diamater can be used to evaluate the prognosis of early NSCLC after radiotherapy.

Human kallikrein-1 (hK1) gene was cloned from kidney tissues cDNA, it was inserted into the plasmid pPICZalphaA, then the yeast expression vector pPICZalpha-hK1 was constructed. After transformed into Pichia pastoris host X33, high-level expression transformants were screened by escalating the concentration of Zeocin (from 500 to 700 microg/mL) of YPD plate and medium. When temperature was 30 degrees C, pH 6.0 with induction duration of 64 hours in the 30 L fermenter, the highest yield can reach about 6500 u/L (1.25 g/L). The variation of glycosylation resulted in two kinds of molecules, i.e. rhK1-H with a heavy molecular weight and rhK1-L with a light one. rhK1 was purified from the supernatant through Phenyl hydrophobic interaction, Cu(2+)-charged Chelating and Anion-exchange chromatography. 0.28 g rhK1-H and 0.62 g rhK1-L can be purified from one liter supernatant. The yield recovery was 72% with a purity of > 96%. So far our yield of rhK1 is superior than known recombinant expression method reported by other researchers.
Amino Acid Sequence ; Base Sequence ; Chromatography, Ion Exchange ; methods ; Genetic Vectors ; genetics ; Humans ; Kidney ; metabolism ; Molecular Sequence Data ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Tissue Kallikreins ; biosynthesis ; genetics

The gentamicin sulfate carried calcium sulfate (GSCS) implants were fabricated by the coagulating method, and the release rate of the gentamicin tested by UV-spectrometer and the absorbing rate of the calcium sulfate carrier in vitro were studied. The release patterns of two types of GSCS were compared. The trend of daily weight loss of GSCS was found being similar to that of pure calcium sulfate, which suggested that the gentamicin part has little effect on the absorbing pattern of calcium sulfate. The release rate of gentamicin is controlled by the erosion rate of calcium sulfate, so GSCS with different amount of gentamicin has the same release patterns. The DRP value of ED is higher than that of CO during the early stage, while the DRP value of ED is lower than that of CO during the late stage. The GSCS samples were implanted into the defect mold on the radii of the rabbits to investigate the potential for the use of GSCS implants as bone fillers, and the results revealed that new bone had been induced in a great part of the defect at 14 weeks after operation.
Animals ; Anti-Bacterial Agents ; administration & dosage ; pharmacokinetics ; Bone Regeneration ; drug effects ; Calcium Sulfate ; administration & dosage ; Gentamicins ; administration & dosage ; pharmacokinetics ; Implants, Experimental ; Rabbits ; Radius Fractures ; therapy