Serum metabolomic profiling of acute complete spinal cord transection in macaques / 中国组织工程研究

BACKGROUND:Non-targeted metabolomic profiling is used to uncover metabolic changes and to identify relationships between metabolites and spinal cord injury,which contributes to further understanding the pathophysiological process and mechanisms of secondary spinal cord injury.OBJECTIVE:To detect and analyze the serum metabolite changes after complete spinal cord transaction in macaques,explore its relationship with the pathophysiological progress of spinal cord injury,and screen the potential biomarkers.METHODS:Five adult macaques were selected,in which the models of complete spinal cord transaction were established.The serum metabolic features were detected using a non-targeted metabolic profiling strategy based on gas chromatography time-of-flight mass spectrometry at 1 day before modeling,3 hours (superacute phase) and 3 days (acute phase) after modeling.After compared with the spectrometry profiling,recognizing the metabolites,searching for differential metabolites and the related metabolic pathways,the pathophysiological process and mechanisms were analyzed.RESULTS AND CONCLUSION:Three hundred and fifty-eight chromatographic peaks were obtained for subsequent data analysis.Fourteen metabolites,including low-molecular-weight organic acid,amino acids,fatty acid and carbohydrate,were identified as differential metabolites.To conclude,the acute phase of complete spinal cord transection is closely related to some metabolic pathways,such as amino acid metabolism,tricarboxylic acid cycle and pyruvate metabolism.

Renal Phospholipase A2 Receptor and the Clinical Features of Idiopathic Membranous Nephropathy / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>According to the renal phospholipase A2 receptor (PLA2R) immunohistochemistry, idiopathic membranous nephropathy (iMN) could be categorized into PLA2R-associated and non-PLA2R-associated iMN. This study aimed to examine whether the non-PLA2R-associated iMN had any difference in clinical features compared with PLA2R-associated iMN.</p><p><b>METHODS</b>A total of 231 adult patients diagnosed as iMN were recruited to this retrospective study. Renal PLA2R expression was examined by immunofluorescence. Among these patients, 186 (80.5%) with complete baseline clinical data were used for further study. Urinary protein excretion, serum albumin, and creatinine were analyzed. For those patients with follow-up longer than 1 year, the relationship between PLA2R and response to immunosuppressants were analyzed. The t-test was used for parametric analysis and the Mann-Whitney U-test was used for nonparametric analysis. Categorical variables were described as frequencies or percentages, and the data were analyzed with Pearson's Chi-square test or Fisher's exact test.</p><p><b>RESULTS</b>Of the 231 iMN patients, 189 showed renal detectable PLA2R expression (81.8%). The baseline serum creatinine, serum albumin, and urine protein excretion were not significantly different between PLA2R-associated (n = 145) and non-PLA2R-associated iMN patients (n = 41). However, about 1/3 of the non-PLA2R-associated iMN had abnormal serological tests, significantly more common than PLA2R-associated iMN (31.7% vs. 8.3%, P = 0.000). The non-PLA2R-associated iMN had lower C4 levels compared with PLA2R-associated iMN (P = 0.004). The non-PLA2R-associated iMN patients also showed a better response to immunosuppressants (complete remission [CR] 42.9%; partial remission [PR] 14.3%) compared with PLA2R-associated iMN (CR 3.2%; PR 48.4%, P = 0.004) at the 3rd month.</p><p><b>CONCLUSIONS</b>There were no significant differences in serum creatinine, albumin, and urine protein excretion between PLA2R-associated and non-PLA2R-associated iMN, while the non-PLA2R-associated iMN patients showed more abnormal serological tests. The non-PLA2R-associated iMN seemed to respond more quickly to the immunosuppressive therapy compared with PLA2R-associated iMN.</p>

Role of HCN channels in the nervous system: membrane excitability and various modulations / 中国应用生理学杂志

Hyperpolarization-activated and cyclic nucleotide-gated (HCN) channels, distributing in a variety of tissues, especially in excitable cells such as heart cells and many kinds of neurons, have an important role in the modulation of heart rate and neuronal excitability. Different from typical voltage-gated sodium channels and potassium channels, HCN channels were evoked inward currents when the cell was hyperpolarized. More and more recent studies have disclosed that HCN channels play important roles in the nervous system, which were linked with its special electrophysiological features as well as its regulatory effect on the cellular membrane excitability. HCN channels could be modulated by many factors including both extracellular molecules and intracellular signaling cascades, which made its functions complicated in the different condition. Based on its role, HCN channels are presumed to be a promising target for chronic pain and brain disorders. In this paper, we will focus on the advancement of roles of HCN channels in the neural system as well as its complex modulator factors.

Aplastic anemia with macrocytic anemia: a study based on long-term follow-up / 中华血液学杂志

<p><b>OBJECTIVE</b>To elucidate the clinical features, response rate, prognosis and clonal evolution of aplastic anemia (AA) with macrocytic anemia (mAA).</p><p><b>METHODS</b>The clinical features at initial diagnosis and data in follow up of mAA hospitalized from January 2000 to October 2011 were analyzed retrospectively.</p><p><b>RESULTS</b>(1) Of 153/568 (26.9%) cases of mAA at initial diagnosis, 114(74.5%)were non-severe AA (NSAA), 39(25.5%)severe AA (SAA) and 0 very severe AA (VSAA), while the proportion was 16.2%, 45.2%, and 38.6% in 376 normocytic anemia AA (nAA), and the difference is statistically significant(χ(2) = 181.390; P = 0.000). The median age of mAA was significantly higher than that of nAA \[30(4 - 70)years vs 19 (3 - 68) years, P = 0.001\]. (2) There were no statistical difference in hemoglobin, absolute neutrophil count (ANC), platelet count (PLT), response rate after 6 months treatment and overall survival (OS) between mAA and nAA grouped in SAA and NSAA respectively. In SAA, the reticulocyte count (Ret) of mAA was significantly higher than that of nAA \[23.90(2.99 - 61.00)×10(9)/L vs 13.1(0 - 70.60)×10(9)/L, P = 0.000\] and the proportion of erythroid cells in bone marrow of mAA was also higher \[23.5 (0 - 58) vs 14.5 (0 - 65), P = 0.043\], while they did not differ significantly in NSAA. (3) The proportion of AA with PNH clones or abnormal cytogenetics did not differ significantly in mAA and nAA groups before treatment. The incidences of AA evolved to PNH in mAA and nAA was not statistically significant (7/153 vs 9/376, χ(2) = 1.099, P = 0.294) and so was the incidence of evolution to MDS/AML(3/153 vs 13/376, χ(2) = 0.399, P = 0.528).</p><p><b>CONCLUSION</b>In presented with macrocytic anemia at initial diagnosis of AA, higher proportion of NSAA, elderly age, higher Ret and proportion of erythroid cells are features, but being no statistical difference in the response rate, OS, and proportion of clonal evolution.</p>

Transplantation of the cutaneous fibular flap combined with anterolateral thigh flap for the repair of complex tissue defect of the leg / 中华显微外科杂志

@#Objective To explore the clinical application and effect of the transplantation of the cutaneous fibular flap combined with anterolateral thigh flap for the repair of complex tissue defect of the leg. Methods The cutaneous fibular flap combined with anterolateral thigh flap in series connection or parallel connection transfer were applied to repair complex tissue defect of the leg in 36 cases. 10 cases were fresh non-infectious wound 26 cases were delayed infectious wound. The area of wound ranged from 25 cm × 18 cm to 45 cm × 13 cm (36 cm × 16 cm on average). The area of anterolateral thigh flap ranged from 12 cm × 13 cm to 32 cm × 18 cm. The area of the cutaneous fibular flap ranged from 2.0 cm × 1.5 cm to 18.0 cm × 16.0 era. The length of fibular transplantation ranged from 10 cm to 24 cm. 30 cases were combined in parallel connection transfer, 6 cases were combined in series connection transfer, 5 cases were repaired in emergency, 5 cases were repaired in subemergency, 26 cases were repaired in delay. Results All cases were successfully repaired in 36 cases.35 cases were followed up. A mean follow-up was 29 months. Arterial crisis occurred in 1 case, venous crisis occurred in 2 cases 34 flaps survived completely and 2 cutaneous fibular flap survived partially in parallel connection which were later healed by skin transplantation.32 cases were healed in first stage, 4 cases were healed in second stage, (healing time ranged from 12 to 18 days), Bone healing time ranged from 3 to 6 months in fibula transplantation. The Enneking score system was applied to evaluate the leg function. Of the 35 cases, the mean scores was 26 (their scores ranged from 23 to 28).The functions of all supplied regions were not found malfunctional. Conclusion Transplantation of the cutaneous fibular flap combined with anterolateral thigh flap is an optimal method to repair the complex tissue defect of the leg.

Serum proteomic variation study in patients with Crohn disease / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To search differentially expressed proteins in serum of patients with Crohn disease.</p><p><b>METHODS</b>Serum protein samples from 4 patients with Crohn disease and 8 healthy adults were recruited cross-labeled with variant CyDye, and then followed by two-dimensional differential in-gel electrophoresis (2-D DIGE), image analysis, and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).</p><p><b>RESULTS</b>The 2-D electrophoresis results were compared between the Crohn disease patients and the healthy adults. The spot 1058 expression in serum of Crohn disease patients increased by 1.68 folds as compared with healthy adults (P<0.05). The protein was identified as haptoglobin by mass spectrometry.</p><p><b>CONCLUSION</b>Up-regulating expression of haptoglobin in serum of Crohn disease patients may play a role in disequilibrium of immunity system.</p>

Treatment for ipsilateral fractures of the femoral neck and shaft / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the curative effect of different internal fixation for ipsilateral fractures of the femoral neck and shaft.</p><p><b>METHODS</b>By retrospective study of 27 patients who sustained ipsilateral femoral shaft and neck fractures from June 1993 to March 2004. There were 22 male and 5 female, with an average age of 35 years (range in 14 to 65 years). The femoral neck and shaft fractures were stabilized with dynamic hip screw system (DHS) in 3 cases,with dynamic compression plate and cannulated lag screw in 12 cases, with constructive nail in 8 cases, with antegrade intramedullary locking nail and cannulated lag screw in 4 cases. There were 13 cases used of the temporary fixation of Kirschner wire before the pexia of the femoral neck fractures.</p><p><b>RESULTS</b>All of the patients were followed up for 36 to 75 months, with an average of 44 months. The average healing period of femoral neck fracture was 4.5 months in 25 cases, nonunion of femoral neck fractures in 2. The average healing period of femoral shaft fracture was 6 months in 27 cases. In 14 cases that not using temporary fixation of femoral neck with Kirschner wire, there were nonunion of femoral neck in 2 and slight coxa vara in 3.</p><p><b>CONCLUSION</b>There are a wide choice of internal fixation method for treatment of ipsilateral fracture of the femoral neck and shaft, the fixation with dynamic compression plate and cannulated lag screw is a handy method. It would be avoided to the replacement and trauma, emporary fixing the fracture with Kirschner wire before the pexia of the femoral neck fracture.</p>

Detection of serum biomakers of osteosarcoma by proteomic profiling / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To detect differentially expressed proteins in serum of patient with osteosarcoma.</p><p><b>METHODS</b>8 serum protein samples were recruited (4 cases of osteosarcoma and 4 cases of normal adults), cross-labeled with variant CyDye, followed by two-dimensional differential in-gel electrophoresis (2-D DIGE), image analysis, and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).</p><p><b>RESULTS</b>24 protein spot-features were significantly increased, and 34 were significantly decreased in the serum from patients with osteosarcoma relative to the controls. The mass spectrometry analysis revealed 18 unique proteins that were increased, and 25 unique proteins decreased in the serum of patients with osteosarcoma. Gelsolin was down-regulated in osteosarcoma, and Western blotting also confirmed a decreased level of gelsolin in the serum of patients with osteosarcoma.</p><p><b>CONCLUSION</b>Our results indicate that gelsolin may have great potential as a biomarker of osteosarcoma and as a potential target for therapy. These preliminary data suggest that incorporation of more samples and new datasets will permit the identification of serum biomarkers for osteosarcoma.</p>

GPI-1046 stimulates chicken dorsal root ganglion neurite outgrowth in the presence of nerve growth factor at low concentration in vitro / 生理学报

The purpose of this investigation was to re-evaluate the neurotrophic effect of GPI-1046 on neurite outgrowth in vitro. GPI-1046 was synthesized and identified with mass spectrometry, nuclear magnetic resonance and elemental analysis. Chicken dorsal root ganglions (DRGs) were removed and divided into three groups: (1) The DRGs were cultured in DMEM containing different concentrations of GPI-1046; (2) The DRGs were cultured in DMEM containing nerve growth factor (NGF) alone at 0.8 and 8 ng/mL, respectively; (3) The DRGs were cultured in DMEM containing both different concentrations of GPI-1046 and NGF at 0.8 ng/mL. The results showed that GPI-1046 alone could not stimulate chicken DRG neurite outgrowth; however, GPI-1046 stimulated DRG neurite outgrowth only in the presence of NGF at low concentration in the culture medium.

Effect of vascular endothelial growth factor antisense oligonucleotide on human leukemic cell line HL-60 / 中国实验血液学杂志

This study was aimed to investigate expression of vascular endothelial growth factor mRNA (VEGF mRNA) and its relationship with leukemic cell apoptosis after VEGF antisense oligonucleotide (VEGF ASODN) transferred into HL-60 cells. The phosphorothiate VEGF ODN was transferred into HL-60 cells in vitro by using cation poly mediated method, the inhibitory rate of cell proliferation was assayed by MTT, expression of VEGF mRNA was measured by RT-PCR, cell apoptosis was detected by cell morphology observation, DNA agarose gel electrophoresis and flow cytometer (FCM). The results showed that difference of the inhibitory rate of cell proliferation and the relative expression of VEGF mRNA between ASODN group and MSODN or control groups under the same condition (p < 0.05) was statistic significant, but no significant difference (p > 0.05) was found between MSODN and control. The number of clusters of cells in ASODN group decreased; the morphology features of apoptotic cells involved cell shrinking, more granulation in cytoplasm, nuclear contracting and many fragments of cells. In MSODN and control groups, however, cells were plump and clear, and grow healthly. The result of electrophoresis revealed DNA ladder in ASODN group, while only one band of DNA in control groups. The rate of cell apoptosis was 19.46% in ASODN group with a significant difference as compared with MSODN groups and control (p < 0.05). The rate of HL-60 cell apoptosis in combination of VEGF ASODN with VP16 was significantly higher than that in VP16 alone (p < 0.05) and showed time- and dose- dependence. It is concluded that VEGF ASODN can down-regulate expression of VEGF mRNA of HL-60 cells, induces the apoptosis, inhibits the proliferation of HL-60 cells and enhances VP16-induced apoptosis in HL-60 cells, the VEGF ASODN in combination with VP16 shows additive effect.

Effect of prostaglandin E1 on the expression of tissue inhibitor of metalloproteinase-1 in experimental liver fibrosis rats / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of prostaglandin E1 (PGE1) on the expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) in experimental liver fibrosis rats.@*METHODS@#The liver fibrosis model was established by carbon tetrachloride. Rats were divided into a control group and PGE1-treated group. The pathological changes of the liver tissue from the two groups, the semi-quantitative analysis of hepatitic activity in HE stain sections, the pathological image quantitative analysis of the fibrosis degree, TIMP-1 positive cells, and the content of collagen were synthetically analysed.@*RESULTS@#The mark changes of liver pathology in HE stain sections were that the degree of hepatitic activity in the PGE1-treated group was obviously lower than that in the control group (P < 0.05). The fibrosis degree, TIMP-1 positive cells and the collagenous fibers decreased in the PGE1-treated group (P <0.05).@*CONCLUSION@#PGE1 has an anti-hepatofibrosis effect in the experimental rats, the inflammation of liver is light, and the proliferation of collagenous fibers can be restrained, whose mechanism is probably associated with the suppression of TIMP-1 expression caused by PGE1.

An investigation on the division of neuronal PC12 cells induced by nerve growth factor / 生理学报

Neuronal PC12 cells induced by nerve growth factor (NGF) have been considered to be postmitotic and lack the ability to divide. However, in this study, we not only detected DNA synthesis but also observed cell division in some morphologically differentiated neuronal PC12 cells bearing long neurites. More interestingly, in addition to the division of perikaryon, the neurites located on the division site of the cell membrane also divided into two parts and were allocated to the two daughter cells. These results demonstrate that the morphologically differentiated neuronal PC12 cells still retain the ability to divide. This is the first report that neuronal PC12 cells as well as their neurites can divide.

cDNA microarray analysis of spinal cord injury and regeneration related genes in rat / 生理学报

The acute traumatic spinal cord injury (SCI) is a commonly seen and severe case in clinic. However, the repair and regeneration of injured spinal cord is limited. This is likely due to that different kinds of factors are involved in regeneration after SCI. In the present study, we used complementary DNA microarray consisting of 4 041 specific probes from rat to identify genes that were differentially expressed after SCI. The animals were subjected to complete transection injury of the thoracic spinal cord (T8-T9). Sham operated animals received only a laminectomy. Four and a half days later, rat spinal cord was dissected out for total RNA isolation. The fluorescent (Cy3 and Cy5) labeled probes were prepared and hybridized to the microarray. Genes that showed 2-fold difference in SCI tissue were identified. Sixty-five up-regulated genes consisted of 21 known genes, 30 known expressed sequence tags (ESTs) and 14 unknown genes. Seventy-nine down-regulated genes comprised 20 known genes, 42 known ESTs and 17 unknown genes. In 41 differentially expressed known genes, 5 up-regulated genes, i.e., tissue inhibitor of metalloproteinase 1 (Timp1), transgelin (Tagln), vimentin (Vim), Fc gamma receptor, cathepsin S (Ctss), and 3 down-regulated genes, i.e., stearyl-CoA desaturase, coagulation factor II (F2), endosulfin alpha (Ensa), were further confirmed by reverse transcription polymerase chain reaction (RT-PCR). These genes may play a role in the response to tissue damage or repair following SCI and characterization of them might be helpful to elucidate the molecular mechanisms of spinal cord injury and regeneration.

Successful treatment of agammaglobulinemia by HLA-mismatched unrelated cord blood stem cell transplantation--the first case report / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate cord blood stem cell transplantation (CBT) in the treatment of X-linked agammaglobulinemia, and observe the courses of the hematopoietic and immune reconstitution.</p><p><b>METHODS</b>A 14-year-old male patient with agammaglobulinemia received CBT from a 1/6 HLA-mismatched unrelated cord blood. The conditioning regimen was Bu/Cy/anti-CD3 antibody. CsA was given together with MMF and MTX for prophylaxis of GVHD. The patient received 0.42 x 10(8) nucleated cells/kg, containing 0.35 x 10(6) CD34(+) cells/kg.</p><p><b>RESULTS</b>The recipient showed hematopoietic reconstitution on day 30 post-transplantation when ANC was 0.5 x 10(9)/L and BPC 20 x 10(9)/L. Sex chromosome analysis showed engraftment (donor 46, XX/recipient 46, XY = 4:1) on day 45. The recipient's blood group changed from AB to O, IgG from 1.1 g/L to 3.5 g/L, sex chromosome from 46, XY to full 46, XX, and mature B cells in peripheral blood from 0 to 5% on day 100, indicating immune reconstitution. At the last follow-up of 360 days, the patient without acute or chronic GVHD showed normal hemogram and myelogram, IgG 13.5 g/L and 10% mature B cells in peripheral blood, indicating the hematopoiesis and immune persistent reconstitution. No acute or chronic GVHD was developed.</p><p><b>CONCLUSION</b>This is the first case report of successful treatment of X-linked agammaglobulinemia by HLA-mismatched unrelated CBT.</p>

Induced-division of neurons derived from neural stem cells / 生理学报

In order to explore if mature neurons derived from neural stem cells have the potentiality to divide, we utilized the chemical digestion method to disperse the adult rat brain tissue into single cells, and culture them in serum-free medium. After being cultured for about eight days in vitro, the neural stem cells were induced to differentiate into neurons. The neurons were further induced to divide. Utilizing the method of serial photograph and NF-160 immunocytochemistry, the processes of division of some neurons were recorded. At the same time, PCNA+NF-160 (or Chat, GABA, GAD) double label were used to investigate if the dividing-neurons were mature ones. After the neural stem cells were induced to differentiate in vitro for eight days, they possessed the shape and character of mature neurons. The differentiated neuron had a big nucleus and one or two distinct nucleolus in the nuclear. Within the perikaryon,there were a large amount of dense and Nissl body-like structure. Several long processes emerged from various locations of the cell body. Then, EGF and bFGF were added into the medium to induce division. After two days of induced-division, neuron-like cells were observed to divide; moreover, the number of neuron-like cells in the region increased continually. Immunocytochemistry demonstrated these cells were NF-160-positive. Serial photographs of dividing-process of neuron-like cells were obtained and their daughter cells were also NF-160-positive. After PCNA+NF-160 (or Chat, GABA, GAD) double label, some cells showed brown cell plasma and black nucleus. The above-mentioned results indicate that neurons, which were previously thought to be end-differentiated, can be re-called into cell cycle under appropriate conditions. Mature neurons still have the potential to divide, proliferate and self-renew.

Comparison of the two-dimensional electrophoresis maps of rat spinal cord protein extracted by two different solution systems / 军事医学科学院院刊

Objective:To compare the two-dimensional electrophoresis(2-DE) maps of rat spinal cord protein extracted by two different solution systems.Methods: Adult rat spinal cord protein was precipitated with 10% trichloracetic acid in acetone and resuspended in 8 mol/L urea plus 4%CHAPS (A solution) or, 5 mol/L urea, 2 mol/L thiourea, 2%CHAPS plus 2%SB3-10 (B solution). One hundred and fifty micrograms of protein was loaded on 18 cm IPG strip holder and run isoelectric focusing electrophoresis as the first dimension, then horizontal SDS-PAGE as the second dimension. Protein spots were visualized by silver stain.Results:There were 1 059 and 1 023 protein spots in each map, of which 790 spots were matched in two maps. There were 269 and 233 spots exclusively extracted by A and B solutions, respectively. Taken together, 1292 different spots were totally obtained by A and B solutions.Conclusion: Integrating protein spots extracted by different solution systems is beneficial for achieving intact 2-DE map of tissues.

Analysis of Clinical and Hematologic Features in 22 Patients with Metastatic Carcinoma of Bone Marrow / 中国实验血液学杂志

The clinical and hematologic features in 22 patients with metastatic carcinoma of bone marrow were observed and analyzed. Morphology of bone marrow cells, bone marrow biopsy and other accessory examinations were performed. The primary or cardinal symptoms of metastatic carcinoma of bone marrow included anemia (17 cases, 77.3%), ostealgia (10 cases, 45.5%), fever (8 cases, 36.4%), hemorrhage (4 cases, 18.2%) and complicated hemolytic anemia (4 cases, 18.2%). The primary carcinomas, diagnosed by pathologic and accessory examinations, include gastric carcinoma (6 cases, 27%), lung cancer (3 cases, 13.6%), ovarian cancer (2 cases, 9%), mammary cancer, prostatic carcinoma, osteocarcinoma and metastatic malignant melanoma (1 case, respectively), and unknown primary lesion (7 cases, 31.8%). The hematologic features were decrease of hemoglobin (17 cases, 77.3%) and blood plate count (16 cases, 72.7%), leukocytosis (11 cases, 50%), immature leukocytes (14 cases, 63.6%) and erythrocytes (9 cases, 40.9%) seen on the peripheral blood smear, and reticulocytosis (4 cases, 18.2%). Masses of metastatic carcinoma cells can be frequently seen at two sides and tail of bone marrow smear. Bone marrow biopsy of 8 cases demonstrated the infiltration of carcinoma cells with nest-like distribution in the bone marrow cavity. Examination of MRI in 6 case showed destruction of bone and corpus vertebra and abnormal signal focus. Bone marrow biopsy could contribute to improve the accuracy of diagnosis and determine the origin of primary carcinoma. MRI plays an important role in diagnosis of metastatic carcinoma in bone marrow.