BACKGROUND: Combined use of multiple interventions for different targets play superimposed or synergistic effects,which has become the current idea for spinal cord injury treatment.OBJECTIVE: To investigate the synergistic effects of low doses of 17-β estradiol combined with bone marrow mesenchymal stem cells (BMSCs) transplantation on the recovery of motor function and inflammatory reactions after spinal cord injury in rats.METHODS: The 10 of 70 male Sprague-Dawley rats served as sham group in which the spinal cord was only exposed but with no treatment, and the rest 60 rats were used to make animal models of spinal cord injury using modified Allen's method and then randomized into four groups (n=15 per group): model, estrogen, stem cell and combined treatment groups. Rats in the stem cell and combined treatment groups were given BMSCs transplantation at injured side; rats in the estrogen and combined treatment groups were given intramuscular injection of 17-β estradiol at 1 and 24 hours after modeling. At 1, 3, 5 and 7 days after modeling, rat functional recovery was evaluated by the Basso, Beatlie, Bresnahan score. The expressions of interleukin-1β and tumor necrosis factor-α in the injured spinal cord were detected by ELISA at 6, 12, 24, and 72 hours after modeling. Apoptosis in nerve cells was observed using TUNEL staining. RESULTS AND CONCLUSION: The Basso, Beatlie, Bresnahan scores were declined significantly after modeling,increased at 5 and 7 days after stem cell transplantation, estrogen treatment or their combined treatment (P < 0.05),especially in the combined treatment group (P < 0.05). The levels of interleukin-1β and tumor necrosis factor-α were elevated gradually after spinal cord injury (P < 0.05), but the levels decreased significantly at 12 and 24 hours in stem cell,estrogen and combined treatment groups (P < 0.05), and this decrease trend was more significant in the combined treatment group compared with the stem cell and estrogen groups (P < 0.05). At 72 hours after modeling, the rate of TUNEL positive cells was highest in the model group (P < 0.05) and lowest in the combined treatment group (P < 0.05).To conclude, the combined use of low doses of 17-β estradiol and BMSCs transplantation can facilitate the recovery of motor function after spinal cord injury by effectively inhibiting apoptosis in nerve cells.

Objective To investigate plasma levels of high-sensitivity C-reactive protein (hs-CRP) in patients with coronary heart disease (CHD) and relationship between hs-CRP levels and cardiac function. Methods The serum hs-CRP concentrations in 894 CHD patients diagnosed by coronary angiography and 141 healthy controls were measured by particle enhanced immunoturbidimetric assay. Results The median of hs-CRP levels was 1.70 (0.13-19.53 ) mg/L and abnormal ratio was 37.6% (336/894), that were significantly higher in CHD group than healthy controls [0. 99(0. 13-19. 53) rag/L, 7. 1% (10/141) ] (Z=-6. 476,X<'2> = 50. 882, P <0.01 ). The median of hs-CRP levels was 5.35 (0. 18-19. 10) mg/L and abnormal ratio was 63.9% (92/144), that were much higher in acute myocardial infarction (AMI) group than in old myocardial infarction (OMI) group [2.27 (0.13-19.19) mg/L, 43.7% (129/295) ] (Z = -3.353 ,X<'2> = 15. 732, P <0. 01 ). The median of hs-CRP levels was 1.45 (0.19-19.53) mg/L in unstable angina pectoris(UAP) group and abnormal ratio was 29.1% (73/251), that were was higher in stable angina pectoris (SAP) group [1.04 (0.13-16.31 ) mg/L, 20. 6% (42/204) ] (Z=-2.981, P = 0.003;X<'2> = 4.30, P=0.038 ) . Furthermore, Kruskal-Wallis test showed the concentrations of hs-CRP and NT-proBNP was increased along with increment of CHD severity (NYHA functional classification) (X<'2> = 120.185,424.945, P <0.001 ). Multivariate analysis showed that hs-CRP levels positively correlated with NYHA functional classification ( r = 0.328, P <0.01 ) as well as NT-proBNP levels (r=0.413, P <0.01 ). Conclusion Serum hs-CRP level increases along with CHD severity, indicating that hs-CRP may play a certain role in the occurrence and development of CHD.

BACKGROUND: Human transforming growth factor-β1 gene can be used for gene therapy of the degeneration of intervertebral discs, but the key to the experiment is to construct its effective vector.OBJECTIVE: To determine whether or not adult degenerated intervertebrai disc cells cultured in vitro after transfected by eukaryotic expression vector can express the product of human transforming factor-βl, and to provide the experimental basis of gene therapy for intervertebral disc degeneration.DESIGN: Single sample experiment. SETTING: Traumatic Orthopedic Institute of Shandong Province and the Orthopedic Department of Weihai Municipal Hospital.MATERIALS: The experiment was conducted at the laboratory of Traumatic Orthopedic Institute of Shandong Province between October 1999and January 2001. Intervertebral disc samples were from the operated patients with protrusion of irtervertebral disc after the patients were informed.Sample 1 was intervertebral disc at L4/5 from a 30-year-old woman; sample 2 was intervertebral disc at L5/S1from a 30-year-old woman.METHODS: ① Culture of adult degenerated intervertebral disc cells:Samples ex vivo were taken back to the laboratory within 30 minutes; fibrous ring cells and myelin nucleus cells cultured primarily were collected.② Transfection: Cells were put in the 24-well culture plate with 5.5×105cells in each well. Constructed PCI-hTGF-β1 eukaryotic expression vector was used to perform transfection, then transfected PCI group and nontransfected group were set. ③ The expression product of cells transfected for 48 hours was determined with immunohistochemical staining method.MAIN OUTCOME MEASURES: Comparison of absorbance of the positive cell product of eukaryotic expression vector PCI-hTGF-β1 in the primary fibrous ring cells and myelin nucleus cells in each group.RESULTS: ① Sample 1: The absorbance of positive cell product of eukary otic expression vector PCI-hTGF-β1 in the primary fibrous ring cells and myelin nucleus cells was 3.49 and 3.69 times that in PCI group, and 3.55times that in non-transfected group. ② Sample 2: The absorbance of positive cell product of eukaryotic expression vector PCI-hTGF-31 in the primary fibrous ring cells and myelin nucleus cells was 3.56 and 3.46 times that in PCI group, and 3.43 times and 3.33 times that in non-transfected group.CONCLUSION: PCI-hTGF-31, as the effective eukaryotic expression vector in the transfection of transforming growth factor-31 gene to culture degenerated intervertebral disc cells in vitro, can transfect adult degenerated intervertebral disc cells cultured in vitro and obtain the high expression of human transforming growth factorβ1 gene.

Aim To investigate the antiarrhythmic mechanism of taurine-magnesium coordination com-pound on abnormal sodium current channel ( INa ) in-duced by hypoxia-reoxygenation in ventricular myocytes of rats. Methods Single ventricular myocytes were i-solated from each rat heart using enzymatic dissociation through Langendorff retrograde aortic perfusion. Whole-cell patch clamp was applied in voltage clamp mode to record INa both in normal ventricular myocytes and single ventricular myocytes of arrhythmia induced by hypoxia-reoxygenation. Results The peak density of INa was changed from ( 56. 89 ± 2. 07 ) pA/pF to (35. 05 ± 1. 52) pA/pF( n=6, P <0. 01 vs control) by hypoxia-reoxygenation with the INa I-V curve shifting upward. TMCC(200, 400 μmol·L-1)was able to re-store the reduction caused by H/R to (35. 78 ± 1. 95) pA/pF, (41. 52 ± 0. 86) pA/pF, (n=6,P <0. 01) and (48. 34 ± 0. 99) pA/pF(n=6,P<0. 01) respec-tively, but not at 100 μmol·L-1(n=6, P>0. 05), in a concentration-dependent manner, while amioda-rone restored it to (39. 44 ± 1. 24) pA/pF (n=6,P<0. 01 ) . Both high concentration of TMCC and amioda-rone could shift the I-V curve downward. In addition, TMCC and amiodarone could restore the INa inactivation curve and slow down its inactivation, whereas the acti-vation curves showed no significant differences among groups. Conclusion TMCC(200,400 μmol·L-1) could restore the H/R induced INa reduction and shift the I-V curve downward by inhibiting steady-state inac-tivation, which is suggested to be one of the mecha-nisms of the antiarrhythmic effects of TMCC in hypoxia-reoxygenation model.

Objective To discuss MRI diagnositic value and evaluation of the treatment of cervical Brucella spondylitis(BS).Meth-ods MRI data of 39 cases with cervical BS in our hospital were collected.Non-operative treatment,minimal invasive surgery and the open surgery were selected according to the MRI manifestations.The correlation was analyzed by consistensy check,imaging score and clinical effect evaluation.The data were analyzed by SPSS15.0.Results 39 cases of patients were examined by MRI before the treatment.37 cases were reviewed after 6 months treatment and 33 cases were reviewed after 12 months treatment.All of the Kappa values were more than 0.75 by imaging consistency check.MRI of cervical BS possessed characteristic expressions.The treatment effect according to MRI classification before the treatment:(1)drug therapy was performed in 14 cases (Group A);(2)minimal inva-sive surgery was performed in 7 cases (Group B);(3)focus clearance and bone graft were performed in 18 cases (Group C);(4)the clinical effects showed the healing rate in every group at different time point was different and the difference had statistical signifi-cance(P<0.05).The healing rate in Group C was higher than it in other group at 6 months after treatment.There was no significant difference among 3 groups or among different method group(Group C1,C2 and C3)in Group C at 12 months after treatment(P>0.05). Conclusion MRI classification for cervical BS has important value and guiding significance in the selection of clinical conservative treatment,minimal invasive surgery or the open surgery.

Objective To evaluate the utility of MMP9,MPO and sCD40L in detection of the character of coronary artery plaque.Methods From April 2008 to January 2010,118 patients from outpatient of Fu Wai Hospital with chest pain were enrolled.All of them underwent 64 Multiple-detector row spiral computer tomography (64-MDCT),the CT value ＜ 130 Hu patients were enrolled in non-calcified plaque group (71 cases),CT value ≥ 130 Hu patients were enrolled in the calcified plaque group (47 cases).Ninty healthy volunteers were selected as the control group.Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum markers,including MMP9,MPO and sCD40L.Levels of MMP9,MPO and sCD40L of each group were compared.ROC curve was used to evaluate the sensitivity and specificity of the markers in diagnosis of non-calcified plaque.Results MMP9,MPO and sCD40L levels of non-calcified were ( 762.25 ± 368.71 ),[ 844.10 (582.00 - 1220.70) ],(9.37 ± 3.15) μg/L,higher than the healthy control group (342.70 ± 178.53),[426.35 ( 283.20 - 592.00) ],(6.55 ± 2.96) μg/L and calcified plaque group ( 483.12 ± 219.09 ),[ 469.00 ( 302.45 - 723.55) ],( 7.24 ± 2.86) μg/L The difference was statistically significant ( F =42.47,H =50.28,F =17.94,all P ＜ 0.01 ). Areas of MMP9,MPO and sCD40L under the ROC curve to predict non-calcified plaque were 0.854,0.792,0.751 respectively,when the identification threshold for non-calcified plaque were 510.13,537.82,7.05 μg/L respectively,the diagnostic sensitivity was 80％,80％,80％ respectively,and specificity was 80％,67％ and 55％ respectively.Conclusion The serum MMP9,MPO and sCD40L levels can help to determine the　character of coronary plaque.

AIM: To discuss the protective effects and possible mechanisms of 17β-estradiol on human retinal pigment epithelial ( RPE) cells induced by high glucose. ·METHODS: RPE cells were cultured and divided into four groups according to randomized controlled method:blank control group:the cells were treated with 5. 5mmol/L routine glucose medium for processing; high glucose group: cells were treated with 100mmol/L glucose for 12h;17β-estradiol low concentration group: after treated with 10 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h; 17β-estradiol high concentration group: after treated with 100 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h. Cell viability were tested by MTT colorimetric detection. Cells apoptosis were detected by Hochest33258 staining. Intracellular reactive oxygen species( ROS) level were detected by H2 DCFDA staining. Expression of CAT, SOD and MDA were tested by colorimetric detection. · RESULTS: RPE cell activity decreased with the concentration of glucose increased; 17β-estradiol inhibited high glucose-induced cell viability decrease in RPE cells, decreased the apoptosis rate of RPE cells and intracellular ROS generation; besides, 17β-estradiol significantly increased the expression of CAT, SOD and decreased the expression of MDA in RPE cells. ·CONCLUSION: The 17β-estradiol effectively inhibited high glucose -induced RPE cells damage, which provide reliable experimental basis for the treatment of injuries in RPE cells.

OBJECTIVESTo investigate the clinical effect of sequential therapeutic intervention Yupei Qisun [compensating for weakness by invigorating Kidney (Shen) and Spleen (Pi) in advance] in Chinese medicine (CM) and hysteroscopic endometrial mechanical stimulation on the treatment of infertile patients with repeated implantation failure (RIF); and to study the differences in patients' endometrial thickness and type on the day of embryo transfer, serum hormone levels on embryo transfer day and clinical pregnancy outcomes.

METHODSIn the clinical study, 168 frozen-thawed embryo transfer (FET) cycles for couples with RIF conforming to the research protocol were randomly divided into three groups: a CM group with 56 cycles (CM combined with FET), a hysteroscopy group with 55 cycles (hysteroscopic endometrial mechanical stimulation), and a control group with 57 cycles (conventional FET). Differences in endometrial thickness on the embryo transfer day, levels of serum estradiol (E2) and progesterone (P) on the embryo transfer day, the E2/P ratio on the embryo transfer day, biochemical and clinical pregnancy rates, implantation rate, abnormal pregnancy rate and other indices were compared among the three groups.

RESULTSEndometrial thickness, E2 and P levels, and the E2/P ratio on embryo transfer day and other factors had no significant differences among groups. The biochemical pregnancy, clinical pregnancy, and implantation rates of the CM and hysteroscopy groups were significantly higher than the control group (P<0.05), and there were no significant differences between these two groups. The abnormal pregnancy rate had no significant difference among the three groups.

CONCLUSIONSSequential therapy of Yupei Qisun could significantly improve the clinical outcomes of rif-fet cycles, being equivalent to hysteroscopic endometrial mechanical stimulation, and provided a reliable method to treat such infertile couples.

Abortion, Habitual ; therapy ; Adult ; Embryo Implantation ; Embryo Loss ; therapy ; Embryo Transfer ; Endometrium ; pathology ; physiopathology ; Female ; Humans ; Hysteroscopy ; Infertility, Female ; pathology ; therapy ; Medicine, Chinese Traditional ; methods ; Physical Stimulation ; methods ; Pregnancy ; Retreatment ; statistics & numerical data

OBJECTIVETo establish a novel Myc gene transgenic mouse model for spontaneously forming B-lymphoma and assessing its tumorigenesis potential.

METHODSFreshly isolated hematopoietic progenitor cells served as the target for Myc gene transfer mediated by a retrovirus vector. These cells were engrafted into C57BL/6 mice with (60)Co-gamma ray radiation in advance. Tumor latency was measured and the tumor loaded mice were followed for survival time. Tumor was identified with histology and immunostaining. The exogenous Myc gene was detected by Western blot (in liver, spleen, tumor tissue) and flow cytometry (FCM) \[in bone marrow (BM)\].

RESULTSMice BM-infected with mutant Myc gene more readily gave rise to B-cell lymphomas than those infected with wild type Myc gene did Myc gene was expressed highly in BM and tumor tissues but not in liver and spleen.

CONCLUSIONOur model will be a tool in assessing the transforming potential of Myc mutants and in studying cooperation between Myc and other oncogenes. Mutant Myc is more effective than wild-type Myc in promoting B cell lymphomagenesis in mice.

Animals ; B-Lymphocytes ; Cell Transformation, Neoplastic ; Flow Cytometry ; Lymphoma ; Lymphoma, B-Cell ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Retroviridae Infections

To investigate the effects of lanosterol (1), inotodiol (2) and trametenolic acid (3) from Inonotus obliquus against oxidative damage induced by CCl4 in mice, 1, 2 and 3 (20, 10 and 5 mg x kg(-1)) were respectively administered to mice, once a day for 3 days. Then the mice were induced to oxidative damage by CCl4 on the third day 30 min after the administration. The activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX) and the content of malondialdehyde (MDA) and reductive glutathione (GSH) in serum and liver homogenate were determined. And the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and interleukin-6 (IL-6) concentration in serum were detected. The results showed that treatment with compound 1, 2 and 3 could significantly increase the activities of SOD, CAT and GSH-PX in serum and liver homogenate. Furthermore, the content of GSH in serum and liver homogenate increased and MDA content decreased markedly. In addition, compound 1, 2 and 3 could significantly inhibit the activities of ALT and AST in serum, and decrease the IL-6 concentration in serum remarkably. So, compound 1, 2 and 3 can protect mice against oxidative stress injury induced by CCl4. Furthermore, compound 1, 2 and 3 can protect cells from damage through inhibition on ALT, AST and the expression of IL-6.
Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Carbon Tetrachloride ; Catalase ; blood ; metabolism ; Female ; Glutathione ; blood ; metabolism ; Glutathione Peroxidase ; blood ; metabolism ; Interleukin-6 ; blood ; Lanosterol ; analogs & derivatives ; isolation & purification ; pharmacology ; Liver ; metabolism ; Male ; Malondialdehyde ; blood ; metabolism ; Mice ; Oxidative Stress ; drug effects ; Polyporaceae ; chemistry ; Protective Agents ; isolation & purification ; pharmacology ; Random Allocation ; Superoxide Dismutase ; blood ; metabolism ; Triterpenes ; isolation & purification ; pharmacology