OBJECTIVETo study the effect of Buyang Huanwu decoction (BYHWD) inducing angiogenesis on the neuroblast migration from the subventricular zone and its mechanisms after focal cerebral ischemia.

METHODThe middle cerebral artery occlusion (MCAO) was performed to mice for 30 minutes to establish the model. The rats were divided into sham group, model group, BYHWD group and endostatin group. BYHWD (20 g x kg(-1), ig) and endostatin (10 μg, sc) were administered 24 h after ischemia once a day for consecutively 14 days. At 14 d after ischemia, the density of micro-vessel and the number of neuroblasts in the ischemia border zone were determined by immunofluorescence staining. The mRNA and protein expression of cell-derived factor-1 (SDF-1) and brain-derived neurotrophic (BDNF) were examined by real-time PCR and Western blot.

RESULTCompared with the model group, BYHWD significantly increased the density of micro-vessel and the number of DCX positive cells in the ischemia border zone (P < 0.01), and significantly increased the SDF-1 and BDNF mRNA and protein expression (P < 0.01). Compared with BYHWD group, endostatin significantly reduced the density of micro-vessel and the number of DCX positive cells in the ischemia border zone (P < 0.01), as well as the SDF-1, BDNF mRNA and protein expression (P < 0.01).

CONCLUSIONBYHWD could promote the neuroblast migration from the subventricular zone via inducing angiogenesis after cerebral ischemia, the mechanism may be correlated with up-regulating the expression of SDF-1 and BDNF.

Angiogenesis Inducing Agents ; pharmacology ; Animals ; Brain Ischemia ; pathology ; physiopathology ; Brain-Derived Neurotrophic Factor ; analysis ; genetics ; Cell Movement ; drug effects ; Cerebral Ventricles ; pathology ; Chemokine CXCL12 ; analysis ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Male ; Mice ; Mice, Inbred ICR ; Neurons ; drug effects ; physiology

5' and 3' flanking region of ovine BLG were amplified from sheep genomic DNA according to the published whole sequence of ovine BLG and cloned to pGEM-T vector correspondently. By partially sequencing, the sequences of BLG 5' and 3' flanking were the same as that of publication completely. The recombinant structure used to direct exogenous gene especially to express in mammary gland was constructed by joining 4.2 kb 5' flanking with 2.1 kb 3' flanking. In order to assess the efficiency of BLG regulatory elements, green fluorescent protein (GFP) gene as a reporter was fused with BLG construct and transfected the mammary epithelial cells (TD47). Through observation under UV microscope and detection by fluorometer, it is demonstrated that the GFP has been successfully expressed in TD47 cell line. By virtue of direct observation and quantitative analysis, the BLG-GFP construct can be served as a model for the quick assessment of mammary gland expression construct.
3' Flanking Region ; genetics ; 5' Flanking Region ; genetics ; Animals ; Breast ; cytology ; Cell Line ; Cloning, Molecular ; Gene Expression Regulation ; Genes, Reporter ; Green Fluorescent Proteins ; Lactoglobulins ; biosynthesis ; genetics ; Luminescent Proteins ; biosynthesis ; genetics ; Sheep

OBJECTIVETo evaluate the effect of the diameter of abdominal aortic aneurysm (AAA) on endovascular exclusion (EVE) and its results.

METHODSFrom March 1997 to June 2007, 429 AAA patients were treated with endovascular stent-graft exclusion. According to the maximal diameter of abdominal aortic aneurysm, the patients were divided into two groups: group A (diameter < 55 mm, n = 274) and group B (diameter > or = 55 mm, n = 155). The diameter of AAA, involvement of iliac artery, length, diameter and distortion of aneurismal neck in the two groups were recorded and compared retrospectively.

RESULTSPatients in group B were significantly older than group A (73.7 vs 71.1 years, P < 0.05). More patients in group B was complicated with coronary artery disease than those in group A (P < 0.05). The mean diameter of AAA in group A was (46.6 +/- 6.8) mm, and (66.8 +/- 11.2) mm in group B (P < 0.05). Proximal aneurysmal necks were shorter, wider and more tortuous in group B than those in group A (P < 0.05). Extraperitoneal approach, embolism of inner iliac artery and reconstruction of another inner iliac artery and stretch technique were more applied in group B. There were more endoleak during operation in group B and more stent-grafts were used. There was significant difference in morbidity rate between the two groups, while no statistic difference in mortality. And in group B, there were a high rate of endoleak and secondary intervention post operation.

CONCLUSIONSThe diameter of AAA affects EVE and its results. In small aneurysms, EVE carries better outcome than in big aneurysms.

Aged ; Aged, 80 and over ; Aortic Aneurysm, Abdominal ; pathology ; surgery ; Blood Vessel Prosthesis Implantation ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Stents ; Treatment Outcome

OBJECTIVETo investigate the risk factors related to post-asphyxial multiple organ dysfunction (PA-MOD) in neonates.

METHODSA total of 397 neonates with birth asphyxia were enrolled from January 2009 to December 2010.The patients were divided into PA-MOD group (n=179) and non-PA-MOD group (n=218). The risk factors of PA-MOD were retrospectively studied.

RESULTSMultivariate logistic regression analysis showed that severe asphyxia, fetal distress, abnormal labor, and decreased amniotic fluid were the risk factors for PA-MOD among the neonates. Spearman rank correlation analysis showed that the number of the involved organs increased along with the increase of age at admission (P<0.05) and with the decrease of gestational age and birth weight (P<0.05).

CONCLUSIONSThe efforts should be made to enhance perinatal care for neonates, especially for preterm infants and low-birh-weight infants, to decrease the incidence of MOD.

Asphyxia Neonatorum ; complications ; Female ; Humans ; Infant, Newborn ; Logistic Models ; Male ; Multiple Organ Failure ; etiology ; prevention & control ; Risk Factors

OBJECTIVETo investigate the factors upgrading the International Society of Urological Pathology (ISUP) Gleason score using the specimens from preoperative prostatic biopsy and radical prostatectomy.

METHODSA total of 164 patients diagnosed with prostate cancer by biopsy underwent radical prostatectomy. We retrospectively analyzed their age, prostate volume, preoperative PSA level, PSA density (PSAD) , the time interval between biopsy and surgery, the number of positive punctures, positive surgical margin, seminal vesicle invasion, lymphatic invasion, and Gleason scores from biopsy and prostatectomy. We also determined the predictors of Gleason score upgrading by logistic regression analysis.

RESULTSOf the 164 cases analyzed, 95 (57.93% ) showed a consistency between the Gleason score of preoperative prostatic biopsy and that after radical prostatectomy, 55 (33.54% ) increased and 14 (8.52%) decreased after prostatectomy as compared with preoperative biopsy. The prostate volume (P < 0.01) and biopsy score (P < 0.05) were independent predictors of Gleason score upgrading. The risk of Gleason score upgrading was 27 times higher in the patients with the prostate volume ≤ 25 ml and 9 times higher in the 25-40 ml group than in the > 60 ml group (P < 0.05).

CONCLUSIONLow Gleason score of biopsy (≤ 6) and small prostate volume (≤ 40 ml) may be the predictors of Gleason score upgrading after radical prostatectomy.

Biopsy ; Humans ; Male ; Neoplasm Grading ; Organ Size ; Prostate-Specific Antigen ; blood ; Prostatectomy ; Prostatic Neoplasms ; classification ; surgery ; Retrospective Studies ; Risk Factors

BACKGROUNDThe auditory brainstem implants (ABIs) have been used to treat deafness for patients with neurofibromatosis Type 2 and nontumor patients. The lack of an appropriate animal model has limited the study of improving hearing rehabilitation by the device. This study aimed to establish an animal model of ABI in adult rhesus macaque monkey (Macaca mulatta).

METHODSSix adult rhesus macaque monkeys (M. mulatta) were included. Under general anesthesia, a multichannel ABI was implanted into the lateral recess of the fourth ventricle through the modified suboccipital-retrosigmoid (RS) approach. The electrical auditory brainstem response (EABR) waves were tested to ensure the optimal implant site. After the operation, the EABR and computed tomography (CT) were used to test and verify the effectiveness via electrophysiology and anatomy, respectively. The subjects underwent behavioral observation for 6 months, and the postoperative EABR was tested every two weeks from the 1 st month after implant surgery.

RESULTThe implant surgery lasted an average of 5.2 h, and no monkey died or sacrificed. The averaged latencies of peaks I, II and IV were 1.27, 2.34 and 3.98 ms, respectively in the ABR. One-peak EABR wave was elicited in the operation, and one- or two-peak waves were elicited during the postoperative period. The EABR wave latencies appeared to be constant under different stimulus intensities; however, the amplitudes increased as the stimulus increased within a certain scope.

CONCLUSIONSIt is feasible and safe to implant ABIs in rhesus macaque monkeys (M. mulatta) through a modified suboccipital RS approach, and EABR and CT are valid tools for animal model establishment. In addition, this model should be an appropriate animal model for the electrophysiological and behavioral study of rhesus macaque monkey with ABI.

Animals ; Auditory Brain Stem Implants ; Deafness ; surgery ; Evoked Potentials, Auditory, Brain Stem ; physiology ; Female ; Macaca mulatta ; Male

Tetramethylpyrazine (TMP), an effective component of traditional Chinese medicine Chuanxiong, is commonly used to resolve embolism. Its possible therapeutic effect against atherosclerosis has received considerable attention recently. Angiotensin II (Ang II) is highly implicated in the proliferation of vascular smooth muscle cells (VSMCs), resulting in atherosclerosis. The mechanisms of TMP in the proliferation of VSMCs induced by Ang II remain to be defined. The present study was aimed to study the effect of TMP on Ang II-induced VSMC proliferation through detection of nuclear factor-kappaB (NF-kappaB) activity and bone morphogenetic protein-2 (BMP-2) expression. Primary cultured rat aortic smooth muscle cells were divided into the control group, Ang II group, Ang II + TMP group and TMP group. Cells in each group were harvested at different time points (15, 30 and 60 min for detection of NF-kappaB activity; 6, 12 and 24 h for measurement of BMP-2 expression). NF-kappaB activation was identified as nuclear staining by immunohistochemistry. BMP-2 expression was observed through Western blot, immunohistochemistry and in situ hybridization. The results showed that: (1) Ang II stimulated the activation of NF-kappaB. Translocation of NF-kappaB p65 subunit from cytoplasm to nucleus appeared as early as 15 min, peaked at 30 min (P<0.01) and declined after 1 h. (2) TMP inhibited Ang II-induced NF-kappaB activation (P<0.01). (3) Ang II increased BMP-2 expression at 6 h but declined it significantly at 12 and 24 h (P<0.01). (4) BMP-2 expression was also kept at high level at 6 h in Ang II + TMP group but maintained at the normal level at 12 and 24 h. (5) There was no significant difference in NF-kappaB activation and BMP-2 expression between the control group and TMP group. These results indicate that TMP inhibits Ang II-induced VSMC proliferation through repression of NF-kappaB activation and BMP-2 reduction, and BMP-2 expression is independent of the NF-kappaB pathway. In conclusion, TMP has therapeutic potential for the treatment of atherosclerosis.
Angiotensin II ; antagonists & inhibitors ; Animals ; Atherosclerosis ; drug therapy ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; analysis ; antagonists & inhibitors ; Immunohistochemistry ; Muscle, Smooth, Vascular ; cytology ; drug effects ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; NF-kappa B ; analysis ; antagonists & inhibitors ; Pyrazines ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; analysis ; antagonists & inhibitors

OBJECTIVETo investigate the expression profiles of myocardin gene during the differentiation of bone marrow-derived mesenchymal stem cell to smooth muscle cells in the conditional medium combined with a high concentration of fetal bovine serum (FBS).

METHODSMarrow-derived mesenchymal stem cells were isolated and purified from mouse femoral bone and shinbones using differential adherent methods. Cells at the third passage were induced by 20% FBS in conditioned medium, conditioned medium alone, 20% FBS or 10% FBS alone respectively. Mouse aortic smooth muscle cells were cultured as the positive control. Levels of mRNA and protein expression of myocardin and several smooth muscle cells marker genes were determined by immunofluorescence, RT-PCR and Western blot before and 3, 7, 10, 14 d after the induction. The presence of smooth muscle myofilaments was detected by using transmission electron microscope.

RESULTSNaive bone marrow-derived mesenchymal stem cells displayed multiple morphological forms including fusiform, polygon, oval, and micro-spherical, as compared to the single macro-spindle form after the induction. Typical appearance of peak valley was displayed on the 21st day after induction. At the same time, the expression of smooth muscle marker genes was reinforced along with an up-regulation of myocardin expression. Immunofluorescence study showed that the cells expressing myocardin and smooth muscle marker genes such as alpha-SMA and SM-MHC increased. Fluorescence domain of myocardin translocated from cytoplasm to nucleus and the amounts of double positive cells for myocardin with alpha-SMA or SM-MHC also increased. RT-PCR confirmed that the mRNA expression of myocardin increased gradually and remained stabilized after achieving its peak on the 7th day after induction. The expression of smooth muscle marker genes, alpha-SMA and SM22alpha, remained stable on the 10th day of induction. It was also confirmed by Western blot that the protein expression of both myocardin and alpha-SMA were markedly increased during the induction. Finally, transmission electron microscopy revealed the presence of myofilament on the 21st day after induction.

CONCLUSIONSBone marrow-derived mesenchymal stem cells can be effectively induced into smooth muscle-like cells by conditioned medium combined with 20% FBS. Myocardin plays an important role in the differentiation process of bone marrow-derived mesenchymal stem cells to the peripheral smooth muscle cells.

Animals ; Bone Marrow Cells ; cytology ; physiology ; Cattle ; Cell Differentiation ; physiology ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; physiology ; Nuclear Proteins ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics ; metabolism ; physiology ; Up-Regulation

OBJECTIVETo explore the risk factors and prevention strategies of post-operative complications in elderly patients with colorectal cancer.

METHODSData of 107 elderly patients (≥75 years) undergoing surgery for colorectal cancer were collected from January 2006 to December 2009 in the Department of Gastrointestinal Surgery, Peking University People's Hospital. POSSUM and E-POSSUM scoring systems were used to predict post-operative complications. ROC curve and observe/expect(O/E) were used to assess the validity of scoring systems. Logistic regression was used to evaluate the independent risk factors associated with post-operative complications of elderly patients with colorectal cancer.

RESULTSThe predictive complication rates of E-POSSUM and POSSUM in elderly patients with colorectal cancer were 13.9%-86.6%(average, 32.7%) and 19.1%-99.1% (average, 55.5%). The predictive validity of E-POSSUM was better than POSSUM(AUC of ROC: 0.862 vs. 0.576, O/E: 0.771 vs. 0.454), the former was closer to the actual complication rate(25.2%, 27/107). Concurrent diabetes mellitus(P=0.019) and rectal lesion(P=0.005) were independent risk factors associated with surgery-related post-operative complications. Anastomotic leakage was the most common surgery-related post-operative complications. Chronic obstructive pulmonary disease(P=0.026), ASA score(P=0.025), intestinal obstruction(P=0.037) and perforation(P=0.001) were independent risk factors associated with non-surgery-related post-operative complications. Pulmonary infection was the most common non-surgery-related post-operative complication.

CONCLUSIONSThe application of E-POSSUM scoring system can provide more accurate prediction of post-operative complications in elderly patients undergoing surgery for colorectal cancer. Positive interventions should be taken for high-risk patients to prevent post-operative complications.

Aged ; Colorectal Neoplasms ; surgery ; Female ; Humans ; Male ; Postoperative Complications ; etiology ; prevention & control ; Risk Factors

OBJECTIVETo compare the effects and mechanism of blood enriching on mouse model of blood deficiency syndrome induced by cyclophosphamide of albiflorin and paeoniflorin.

METHODAlbiflorin and paeoniflorin were determined by using animal models of blood deficiency syndrome induced by cyclophosphamide. The amount of WBC, RBC, HGB, index of thymus gland and spleen, and the changes of GM-CSF, IL-3 and TNF-α in serum were detected after the treatment.

RESULTCompared with the model group, the amount of WBC in the group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were increased obviously (P < 0.01). The amount of RBC in the group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were increased obviously (P < 0.01, P < 0.001), which did not had a significant difference compared with the same dose. The index of thymus gland in the group of 30 mg x kg(-1) albiflorin was superior to the model group (P < 0.01), the difference was significant compared with the same dose of paeoniflorin (P < 0.05). The GM-CSF in serum in all groups of 30 mg x kg(-1) albiflorin, 15 mg x kg(-1) albiflorin, 30 mg x kg(-1) paeoniflorin and 15 mg x kg(-1) paeoniflorin increased obviously (P < 0.01, P < 0.05, P < 0.01, P < 0.05); The IL-3 in serum in both group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin also increased (P < 0.001). The content of TNF-α in group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were reduced (P < 0.01), which showed the obvious difference compared with the same dose group (P < 0.01).

CONCLUSIONAlbiflorin had the effect of blood enriching by regulating the immune function, same with the paeoniflorin. The probable mechanism of nourishing blood and liver of Paeoniae Radix Alba was not only the better effect of adjusting the content of TNF-α, but also might act synergistically with paeoniflorin.

Animals ; Blood Cells ; drug effects ; Bridged-Ring Compounds ; pharmacology ; Cyclophosphamide ; toxicity ; Glucosides ; pharmacology ; Granulocyte-Macrophage Colony-Stimulating Factor ; blood ; Hematopoiesis ; drug effects ; Interleukin-3 ; blood ; Male ; Mice ; Monoterpenes ; pharmacology ; Tumor Necrosis Factor-alpha ; blood