Objective To isolate fungi from sea sediment and sea water samples collected from the neritic environment in Qingdao, screen their antitumor activity and study on their fermentation condition. Methods The antitumor activity was assayed by flow cytometry in mouse tsFT210 cells. Results The medium added penicillin and streptomycin possesses good selectivity. 207 strains of fungi were isolated and among them, 19 strains showed antitumor activity, and one fungus(Z 83200)pocesses strong cell apoptosis inducing activity.

AIM: To explore the relationship between various mitochondrial (mt) DNA tRNA Leu (UUR) and ND1 gene mutations and type 2 diabetes mellitus (T2DM) among Chinese in Hubei Province. METHODS: PCR restriction fragment length polymorphism (PCR-RFLP) analysis was used to screen point mutations of mtDNA ( 3 243, 3 256, 3 290, 3 316, 3 394, 3 421, 3 426, 3 460, 3 593) in 174 T2DM and 207 healthy controls. Then, DNA sequencing, reverse dot blot hybridization and Genchip were used to compare and confirm mutations. All mutations were analyzed by DNASTAR and Antherprot softwares. RESULTS: In diabetic group, there were 5 carriers (2.9%) of 3 316 G→A (Ala→Thr) mutation, 4 (2.3%) of 3 394 T→C (Tyr→His) mutation, 1 (0.6%) of 3 593 T→C(Val→Ala) mutation, and 1 (0.6%) of 3 618 T→C(Phe→Phe) mutation. Among 3 316 (G→A) mutations , there were more than 1 point mutations in 2 cases, one accompanied with 3 256 C→T(Arg→Arg) and 3 688 G→C (Ala→Pro) mutations, another accompanied with 3 606 A→G(Leu→Leu) mutation. 3 606 (A→G), 3 618 (T→C) and 3 688 (G→C) were novel mutations, GenBank accession number is DQ092356. In controls, only 3 316 (G→A) mutation was found in 1 subject (0.5%). There was significant difference between two groups for 3 394 (T→C) mutation frequencies (P

In order to explore the role of acetylcholine in the pathogenesis of Parkinson's disease (PD), the changes in the concentration of acetylcholine (Ach) in the striatum, the apoptosis of substantia nigra cells, the ultrastructure and the changes of Nissl cells in rats during the morbidity of PD, and the corresponding behaviors in rats with PD were observed. Rat PD model was established by using the modified Thomas method. Eighty-one rats were randomly divided into normal control, sham operation and PD groups and their behavior features were observed at post-operative day (POD) 7, 14 and 21 as three subgroups (n=9 each). The concentration of Ach in the striatum was determined by using high-performance liquid chromatography. The apoptosis of substantia nigra cells was assayed by using TUNEL method. The ultrastructural changes in the substantia nigra were observed under the electron microscopy, and the survival of neurons in the substantia nigra area was examined by using Nissl staining. In PD group at POD 7 to 21, the damage in the substantia nigra area was gradually aggravated, the concentration of Ach, apoptosis rate and turns of rotation were gradually increased, and the number of Nissl cells was gradually reduced over the time as compared with the normal control and sham operation groups (all P<0.05). It was concluded that there exist dynamic changes in Ach concentration, ethology and apoptosis of the substantia nigra cells during the morbidity of PD, suggesting the contribution of apoptosis to the morbidity of PD, and critical role of Ach in the pathogenesis of PD.

OBJECTIVETo study the autophagy activity between rat bone marrow stem cells (BMSCs) neural differentiation in order to explore the mechanism involve in this process.

METHODSBMSCs were passed by 3 generation, then was induced with the revulsant 2% (DMSO) + 200 µmol/L (BHA), NSE expression was detected by immunocytochemical stain, the mRNA expression of autophagy associated genes L3B, Beclinl, Atg5, Atg7, Atg10 were detected by RT-PCR, the autophagy protein LC3B was examined by Western blot and flow cytometry analysis.

RESULTSBMSCs were passed by 3 generation, the purity of BMSCs could reach more than 90%, the morphology of cells were like fibroblasts, after the revulsant 2% DMSO + 200 µmol/L BRA induced, cells were extended long neurites, like nerve cells, positive rate of NSE staining was (83±5) %, RT-PCR results showed that the expression of autophagy associated genes LC3B, Beclinl, Atg5, Atg7 Atg0 were rised after BMSCs neural differentiation, Western blot analysis showed that the LC3B-II protein expression was increased after neural differentiation and the MFI of L3B was highten by flow cytometry.

CONCLUSIONAutophagy is increased after rat BMSC neural differentiation.

Animals ; Autophagy ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats

OBJECTIVETo observe the level of intracellular reactive oxygen species (ROS) in rats with severe burn and pulmonary microvascular endothelial cells (PMVECs) treated with serum of rat with burn injury, and to investigate the relationship between ROS and apoptosis of PMVECs.

METHODS(1) Twenty-four SD rats were divided into sham injury group ( n = 3) and burn group (n = 21) according to the random number table (the same grouping method below). Rats in burn group were inflicted with 30% TBSA full-thickness scald on the back, and rats in sham injury group were sham injured. Blood samples were collected from abdominal aorta at post injury hour 6, 12, 24, 36, 48, 60, 72 respectively from 3 rats of burn group. The serum content of ROS was assayed by ELISA. The same determination was performed in rats of sham injury group. (2) Five rats were subjected to scald injury as above, and burn serum was prepared 24 hours after injury. Another 5 rats without receiving any treatment were used to prepare normal serum. (3) Marginal pulmonary tissue was harvested from 20 SD young rats. Cells were cultured with tissue block method and indentified with immunohistochemical staining. The third passage of PMVECs in logarithmic phase were inoculated in 6-well plates and 12-well plates. PMVECs in both plates were divided into 4 groups: normal serum group, burn serum group, normal serum + MnTBAP group, and burn serum + MnTBAP group, with 3 wells in each group. Cells in the former 2 groups were respectively cultured with special nutrient solution of endothelial cells without serum added with 15% healthy rat serum or 15% burn rat serum. Cells in the latter 2 groups were cultured with the same culture conditions as in the former two groups correspondingly with addition of 100 µmol/L MnTBAP in the nutrient solution. After being cultured for 24 h, the content of ROS in PMVECs in 6-well plates was detected with flow cytometry. The apoptosis of PMVECs in 12-well plates was observed with acridine orange-ethidium bromide staining, and the apoptosis rate was calculated. Data were processed with one-way analysis of variance and LSD-t test.

RESULTS(1) The serum contents of ROS in rats of burn group were respectively (187 ± 21), (235 ± 22), (231 ± 25), (291 ± 20), (315 ±23) nmol/mL at post injury hour 24, 36, 48, 60, 72, which were significantly higher than that in sham injury group [(141 ± 19) nmol/mL, with t values respectively 7. 86, 9. 57, 13. 87, 14.98, 18.40, P values below 0.01]. (2) Primary cells grew slowly and showed a cobblestone appearance. After passages, cells grew with orderly distribution. The positive rate of coagulation factor VIII of cells was (96 ± 5)% , and thus they were identified as PMVECs. (3) In normal serum group, burn serum group, normal serum + MnTBAP group, and burn serum + MnTBAP group, the contents of ROS in PMVECs were respectively 798 ± 40, 1 294 ± 84, 763 ± 59, 926 ± 42 ( F =93.01, P <0.01), and the apoptosis rates of PMVECs were respectively (6.2 ± 1.3)%, (57.3 ± 6. 7)%, (3.7 ± 0. 8)%, (28.7 ± 5. 7)% (F = 224.50, P <0.01) after being cultured for 24 h. Compared with those of normal serum group, the content of ROS and apoptosis rate of PMVECs in burn serum group increased significantly (with t values respectively 10.40 and 49.06, P values below 0.01). The content of ROS and apoptosis rate of PMVECs in burn serum + MnTBAP group were significantly lower than those in burn serum group (with t values respectively 7.48 and 23.94, P values below 0.01).

CONCLUSIONSSerum content of ROS was increased in severely burned rats. Burn rat serum stimulation on PMVECs can lead to the increase of the intracellular ROS and induce apoptosis. However application of MnTBAP can scavenge ROS and reduce the apoptosis induced by burn rat serum.

Animals ; Apoptosis ; Burns ; blood ; therapy ; Endothelial Cells ; pathology ; Enzyme-Linked Immunosorbent Assay ; Lung ; blood supply ; Oxygen ; Rats ; Reactive Oxygen Species ; blood ; Serum ; metabolism

BACKGROUND: Carotid intima media thickness (CIMT) and stiffness are taken as useful surrogate markers of atherosclerosis. In China, the number of elderly patients undergoing hemodialysis has increased year by year, with the increase of dialysis-related cardiovascular events. This study was undertaken to examine carotid stiffness in elderly hemodialysis patients by the ultrasound techniques in order to find out the possible risk factors. METHODS: From January 2006 to February 2010, a total of 87 patients (41 males and 46 females) treated with routine hemodialysis at the 97th Hospital of People's Liberation Army were enrolled in this study. The distensibility coefficient (DC) of the carotid artery was detected by Doppler ultrasonic diagnosis apparatus (Philips HBI5000, frequency 12 MHz) for evaluation of arterial stiffness. Serum albumin, total cholesterol (TC), high density lipoprotein (HDL), low density lipoprotein (LDL), triglyceride (TG), glucose, creatinine, calcium, phosphorus, and intact parathyroid hormone (iPTH) were examined with standard methods. The liner correlation and multiple stepwise regression analysis were used to find correlations between them. RESULTS: In this study, the systolic blood pressure was 153.33±25.98 mmHg, DBP 84.22± 10.39 mmHg, TC 4.39±1.05 mmol/L, TG 1.36±0.72 mmol/L, LDL 2.47±0.77 mmol/L, Cr 889.82± 207.38 mol/L, Glu 5.36±1.87 mmol/L, Ca I 2.00±2.19±0.21 mmol/L, and DC 13.39±5.32×10-3/kPa. DC was associated with age (r =-0.459, P<0.001), SBP (r =-0.527, P<0.001), and serum calcium (r =-0.273, P=0.011). The multiple stepwise regression analysis showed that SBP, age, increased serum calcium level, and diabetes were independent risk factors for decreasing DC. CONCLUSION: Systolic blood pressure, age, increased serum calcium level and diabetes in elderly hemodialysis patients are independent risk factors for increased carotid arterial stiffness.

OBJECTIVETo investigate the dynamic changes that occur in T cell subsets, particularly involving the surface expression of programmed death 1 (PD-1), in response to pegylated (Peg)-interferon (IFN) a-2a therapy in patients with chronic hepatitis C virus (HCV) infection.

METHODSTwenty-five patients with HCV genotype 1b chronic infection and 10 healthy controls were enrolled in the study. All the HCV patients received combination antiviral therapy of Peg-IFNa-2a (180 mug/week) plus ribavirin. At treatment weeks 0 (baseline), 4, 12, 24 and 48, the level of PD-1 protein expression on the surface of total peripheral CD8+ and CD4+ T cells was determined by flow cytometry and the level of PD-1 mRNA expression in peripheral blood mononuclear cells (PBMCs) was determined by reverse transcription-polymerase chain reaction. Independent student's t-test were used to compare mean values between the two groups, repeat measure variance analysis was used to compare mean values among multiple groups, and Pearson's correlation coefficient was used to assess correlation significance.

RESULTSOver the course of antiviral therapy, the proportions of CD4+ T cells and CD8+ T cells, as well as the CD4+/CD8+ ratio, increased (F = 81.23, 39.28, and 7.01 respectively; all P less than 0.01). In contrast, the PD-1 protein expression frequency on CD4+ T cells and CD8+ T cells significantly declined (F = 100.11 and 158.40 respectively; all P less than 0.01). The PD-1-mRNA expression level in PBMCs was: 1.40+/-0.26 at baseline, 1.30+/-0.27 at week-4, 1.14+/-0.18 at week-12, 1.06+/-0.26 at week-24, and 0.83+/-0.25 at week-48 (F = 20.09; P less than 0.01). A positive correlation existed between the PD-1 protein expression frequencies on CD4+ T cells and CD8+ T cells and the HCV RNA load detected at baseline (r = 0.82 and 0.75 respectively; all P less than 0.01).

CONCLUSIONThe ability of Peg-IFN-a-2a-based antiviral therapy to suppress HCV replication may involve reduction of PD-1 protein expression on the surface of CD8+ T cells and CD4+ T cells.

Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; CD4-CD8 Ratio ; Case-Control Studies ; Female ; Hepatitis C, Chronic ; drug therapy ; immunology ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Programmed Cell Death 1 Receptor ; metabolism ; Recombinant Proteins ; therapeutic use ; Ribavirin ; therapeutic use ; T-Lymphocyte Subsets ; metabolism ; Young Adult

In order to explore the role of acetylcholine in the pathogenesis of Parkinson's disease (PD), the changes in the concentration of acetylcholine (Ach) in the striatum, the apoptosis of substantia nigra cells, the ultrastructure and the changes of Nissl cells in rats during the morbidity of PD, and the corresponding behaviors in rats with PD were observed. Rat PD model was established by using the modified Thomas method. Eighty-one rats were randomly divided into normal control, sham operation and PD groups and their behavior features were observed at post-operative day (POD) 7, 14 and 21 as three subgroups (n=9 each). The concentration of Ach in the striatum was determined by using high-performance liquid chromatography. The apoptosis of substantia nigra cells was assayed by using TUNEL method. The ultrastructural changes in the substantia nigra were observed under the electron microscopy, and the survival of neurons in the substantia nigra area was examined by using Nissl staining. In PD group at POD 7 to 21, the damage in the substantia nigra area was gradually aggravated, the concentration of Ach, apoptosis rate and turns of rotation were gradually increased, and the number of Nissl cells was gradually reduced over the time as compared with the normal control and sham operation groups (all P<0.05). It was concluded that there exist dynamic changes in Ach concentration, ethology and apoptosis of the substantia nigra cells during the morbidity of PD, suggesting the contribution of apoptosis to the morbidity of PD, and critical role of Ach in the pathogenesis of PD.
Acetylcholine ; pharmacology ; Animals ; Corpus Striatum ; drug effects ; metabolism ; pathology ; Disease Models, Animal ; Male ; Parkinson Disease ; metabolism ; pathology ; Rats ; Rats, Wistar

OBJECTIVETo report a case of rectal non-Hodgkin lymphoma with concomitant rectal adenocarcinoma.

METHODSClinical records of a 71 years old male patient with rectal non-Hodgkin lymphoma with concomitant rectal adenocarcinoma admitted on May 19, 2010 to the First Affiliated Hospital of Sun Yet-sen University were retrospectively reviewed. Clinical manifestations, diagnosis, and treatment as well as postoperative pathology were summarized.

RESULTSThe preoperative diagnosis of the patient was severe atypical adenomatous hyperplasia with focal carcinogenesis, and the preoperative staging was T2N0-1M0. The patient underwent a Parks procedure (rectal resection and colo-anal anastomosis) and subtotal resection of left lateral liver. The operation was successful, postoperative recovery uneventful. Postoperative pathology showed moderately differentiated tubular adenocarcinoma with deep muscular invasion, and non-Hodgkin lymphoma with marginal zone cell. Both the distal and proximal resection margins were negative and no vascular and neural invasion were seen. Immunohistochemical staining indicated L26(+), Bcl-2(+), Bcl-6(+), CD3(-), CD23(-), CK epithelial cells(+), and M-CEA luminal border(+). The pathological and immunohistochemistry results of liver specimens showed hepatic mucosa-associated marginal zone lymphoma.

CONCLUSIONSRectal adenocarcinoma and lymphoma occurring at the same site simultaneously is extremely rare with unique pathologic features.

Adenocarcinoma ; complications ; pathology ; Aged ; Humans ; Lymphoma, Non-Hodgkin ; complications ; pathology ; Male ; Rectal Neoplasms ; complications ; pathology

OBJECTIVETo evaluate the efficacy of thalidomide in the treatment of juvenile idiopathic arthritis (JIA).

METHODSTwelve children with JIA who did not respond to conventional treatment were administered with thalidomide (2 mg/kg daily). The symptoms, signs, and laboratory test results were compared before and after treatment. The thalidomide-related side effects were observed.

RESULTSThe average dosage of prednisone was reduced from 1.92 ± 0.16 mg/kg•d to 0.49 ± 0.42 mg/kg•d in the 12 patients 6 months after thalidomide treatment (P<0.01). Four patients did not need prednisone treatment any more. White blood cell count, erythrocyte sedimentation rate (ESR), C reactive protein (CRP) and serum ferritin (SF) significantly decreased after treatment in all of 12 patients (P<0.01). Hemoglobin level increased to normal in 8 patients after treatment (P<0.01). The number of affected joints decreased from 5 before treatment to zero to 2 after treatment in patients with polyarticular JIA (P<0.01). Signs of hip involvement and Schober's sign turned negative in enthesitis-related cases. No thalidomide-related side effects were observed.

CONCLUSIONSThalidomide is effective in the treatment of JIA in children who do not respond to conventional treatment.

Adolescent ; Arthritis, Juvenile ; blood ; drug therapy ; Child ; Female ; Humans ; Male ; Prednisone ; therapeutic use ; Retrospective Studies ; Thalidomide ; therapeutic use