Animals ; Cycadopsida ; chemistry ; Down-Regulation ; drug effects ; Female ; Flavones ; isolation & purification ; pharmacology ; Isoproterenol ; Male ; Myocardial Ischemia ; chemically induced ; metabolism ; prevention & control ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

Abstract: Objective　To construct a mouse model of Schistosoma japonicum liver disease induced by direct injection of Schistosoma japonicum eggs through the portal vein and evaluate its effectiveness, in order to provide a new animal model for schistosomiasis liver disease research. Methods　Fifteen 8-week-old C57BL/6 male mice were randomly divided into control group and egg injection group, with 5 in the control group and 10 in the egg injection group. On day -14, 5 000 live eggs were injected into the abdominal cavity of mice, and on day 0, the mice were anesthetized and the abdominal cavity was opened. 5 000 live eggs were injected through the portal vein, and the control group was injected with equal volume of phosphate buffer (PBS). 5 mice in the egg group were killed on day 10 and 30, respectively. The control group mice were killed on day 10, and their serum and liver tissue were collected. Hematoxylin eosin staining (HE) and Masson staining were performed, and alanine aminotransferase (ALT), aspartate aminotransferase (AST), and liver hydroxyproline (HYP) content were detected using a microplate spectrophotometer. Liver fibrosis-related genes, Th1 and Th2 type immune response-related genes were analyzed by real-time fluorescence quantitative PCR (qPCR). Liver injury, egg granuloma and fibrosis, and adaptive immune response were detected to evaluate the effect of portal vein injection of eggs while inducing mouse model of schistosomiasis liver disease. Results　The results showed that significant egg granulomas appeared in the liver of mice after injection of eggs into the portal vein for 10 and 30 days. There was no statistically significant difference in the area of egg granulomas between the 10-day group and the 30-day group (t=0.975, P=0.332). Masson staining and liver hydroxyproline content detection showed significant fibrosis in the liver. The qPCR results showed that, compared with the control group, the expression levels of fibrosis marker genes, such as α⁃Sma (alpha smooth muscle actin), Col1a1 (collagen type Ⅰ alpha 1), and Tgfb1 (transforming growth factor beta 1), were significantly increased (t=6.380, 7.533, 5.314; P=0.002, 0.001, 0.007), and then decreased on the 30th day, with no statistical difference compared to the control group (t=0.940, 1.529, 1.746; P=0.778, 0.543, 0.457). At the same time, the expression levels of Th1 type immune response-related genes, such as tumor necrosis factor alpha (Tnf), interferon gamma (Ifng), and Th2 type immune response-related genes, such as interleukin-5 (Il5), interleukin-13 (Il13), significantly increased 10 days after eggs injection (t=6.163, 4.589, 5.651, 5.367; P=0.003, 0.018, 0.020, 0.009). In addition, there was no significant change in the levels of AST and ALT in the serum of each group of mice (t=0.982, 3.450; P=0.771, 0.074. t=1.164, 0.564; P=0.697, 0.917). Conclusions　A mouse model of schistosomiasis liver disease induced by portal vein injection of worm eggs was constructed. The study provides a new modeling method for studying the mechanism of liver fibrosis in schistosomiasis..

Objective To investigate the diagnostic value of prostate specific antigen (PSA) (fPSA, fPSA/tPSA, PSAD) combined with value of apparent diffusion coefficient (ADC) in diffusion weighted imaging (DWI) and magnetic resonance spectroscopy (MRS) in the diagnosis of prostate cancer (PCa) between total PSA (tPSA) 4- 10 μg/L. Methods From December 2013 to 2015 February , 56 elderly male patients with serum tPSA 4-10μg/L were enrolled. Pathological examination confirmed 26 patients with PCa and 30 patients with benign prostatic hyperplasia (BPH). The levels of tPSA and free PSA(fPSA) , fPSA/tPSA, Prostate specific antigen density(PSAD), ADC value according to the DWI, choline(Cho), creatine(Cre) and citrate(Cit) in MRS were analyzed, and ADC, (Cho+Cre)/Cit combined with PSAD and fPSA/tPSA in PCa diagnosis was analyzed by receiver operating characteristic curve (ROC). Results The level of tPSA in PCa group and BPH group had no significant difference (P >0.05), but the levels of fPSA、fPSA/tPSA、prostate volume, PSAD, (Cho+Cre)/Cit, ADC had significant difference:(1.04 ± 0.14) μg/L vs. (1.22 ± 0.34) μg/L, 0.15(0.14- 0.16) vs. 0.17(0.15-0.18), (41.45 ± 5.70) cm3 vs. (48.70 ± 9.97) cm3, 0.17(0.15-0.18) ng/(ml·cm3) vs. 0.16(0.14-0.17) ng/(ml·cm3), 2.22 ± 0.59 vs. 1.17 ± 0.52, 0.98 ± 0.28 vs. 1.39 ± 0.24, P < 0.01 or < 0.05. The area under the curve of (AUC) fPSA/tPSA+PSAD+(Cho+Cre)/Cit、fPSA/tPSA+PSAD+ADC、fPSA/tPSA+PSAD+(Cho+Cre)/Cit+ADC were 0.932, 0.941 and 0.977, and the AUC of fPSA/tPSA+PSAD+(Cho+Cre)/Cit+ADC was the highest. Conclusions PSAD and fPSA/tPSA and the value of ADC in DWI and MRS can be used in diagnosis of PCa.

Objective:To explore the effect of lipoic acid (LA) on chronic oxidative stress,cytokines and inflammatory gene expression with mice fed with high fat diet (HFD) and whether LA supplementation could prevent development of chronic inflammation.Methods:C57BL/6 mice were randomly assigned to three groups.The control group were administrated with an ordinary diet.The two experimental groups were fed with a high fat diet or high fat plus 0.1% LA.Antioxidants defense index such as SOD,CAT,GSH-Px and MDA were examined after 10 week.Cytokines such as IFN-?,IL-4,IL-6,TNF-? and IL-10 were examined after 10 week,respectively.Gene expression related to oxidative stress and inflammation were confirmed by QRT-PCR.Results:HFD led to potently weaken antioxidant defenses in mice.HFD significantly increased levels of IFN-?,IL-6 and TNF-?,and decreased levels of IL-4 and IL-10 in mice plasma.QRT-PCR results showed an up-regulation of inflammation related genes and a down-regulation antioxidant-related genes.Conclusion:LA is a possibly effective supplementation with HFD,both to prevent from the development of long-term oxidative stress and to attenuate chronic inflammation.

OBJECTIVE:To evaluate the curative efficacy of granulocyte macrophage colony stimulating factor (GM-CSF) as an adjunctive therapy for the treatment of acute myeloid leukemia (AML). METHODS: Randomized controlled trials (RCTs) of GM-CSF in the treatment of AML were retrieved from PubMed,EMBase,Cochrane Library,CBMdisc and CNKI database by computer as well as from the leukemia-related journals manually for a quality evaluation of the methodology and Meta analysis using RevMan5.0 software. RESULTS: A total of 10 RCTs were included,of which,3 RCTs were graded as A,5 as B and 2 as C. The results demonstrated that there were no significant differences of indexes among groups as complete remission (CR) rate,relapse rate,infection rate and length of hospital stay,etc. CONCLUSION: The available clinical research results showed that GM-CSF adjunctive therapy could neither increase CR rate,nor decrease the relapse rate,infection rate or length of hospital stay.

Adolescent ; Adult ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Recovery of Function ; Tibial Fractures ; diagnostic imaging ; physiopathology ; surgery ; Tomography, X-Ray Computed ; Young Adult

According to the sequence of P450 cDNA of Eleutherococcus senticosus, specific primers were designed. Frokaryotic ex pression vector pET30a-P450 was constructed and the prokaryotic expression conditions were optimized. Results showed that the BL21 after being transformed with the recombinant expression vector accumulated the high amount of recombinant protein. SDS-PAGE analysis showed that the recombinant protein was about 53 kDa. The recombinant accumulated the highest amount of recombinant protein af ter IPTG (1 mmol x L(-1)) at 27-37 degrees C for 24 h. Consequently P450 gene of E. senticosus could be expressed successfully by prokaryotic expression vector pET30a-P450. Induction temperature, IPTG concentration, medium type and amount of induction time could all influence the expression of target protein, but the impact strength was different.
Cloning, Molecular ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Eleutherococcus ; enzymology ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plasmids ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism

Objective To learn more about the clinical and laboratory features of childhood paroxysmal nocturnal hemoglobinuria(PNH) and to improve the diagnosis.Methods The clinical and laboratory features of 12 cases of PNH were analyzed,who were diagnosed from January 2000 to November 2004,and the positive responses to treatments were observed.Results 1) The youngest age of onset was 2 years;the disease often manifested with anemia(100%),recurrent infections(50%) and hemorrhages(33%),occurring mainly in skin and mucosa.No patient developed a thrombosis.2) 66.7% of the patients showed peripheral blood cytopenia.Dysplasia of bone marrow was observed in 25% of patients.Fifty percent of them had an increased percentage of erythroid lineage.3) Positive hemolytic tests included urine OB 41.6%,Ham′s test 33.3% and Rous′ test 25%.Glycosyl-phosphatidyl inositol(GPI) deficient cells were found in 100% of the patients.4) 57.1% of patients was improved after being treated with adrenocortical hormone,androgens or cysporin.Conclusions Besides hemoglobinuria,peripheral blood cytopenia were also the common manifestation of PNH.Flow cytometry based immunophenotypic methods for the analysis of CD_(55) and CD_(59) may improve the diagnosis of PNH.J Appl Clin Pediatr,2006,21(3):153-154

OBJECTIVETo optimize Supper Critical CO2 extracting technical (SFE-CO2) methods for extraction of anti-cancer active components of Fig Residues and to investigate the anti-cancer effect of the extract in vitro and in vivo.

METHODThe anti-cancer activity of extracted compound was measured on U937,95D and AGS cancer cells in vitro by MTT method. The anti-cancer effect of the extraction of Fig Residues was studied on mice transplant liver cancer in vivo.

RESULTThe SFE-CO2 condition for extraction of the anti-cancer components of Fig Residues was optimized as follows: granularity was 100, the extraction pressure was 30 MPa, the extraction temperature was 45 degrees C, the extraction time was 6 h and the CO2 flux was 12 L x h(-1); The IC50 of anti-cancer active components of Fig Residues on U937, 95D and AGS cells were 70.125 microg x mL(-1), 127.957 microg x mL(-1), 116.000 microg x mL(-1); The anti-cancer active components of Fig Residues inhibited 49.3% of the transplanted liver cancer in the mice.

CONCLUSIONThe method for extracting the anticancer active components of Fig Residues is stable and reasonable, and the extract from Fig Residues is of the anticancer effect.

Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromatography, Supercritical Fluid ; methods ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Ficus ; chemistry ; Fruit ; chemistry ; Humans ; Inhibitory Concentration 50 ; Liver Neoplasms ; pathology ; Male ; Mice ; Neoplasm Transplantation ; Plants, Medicinal ; chemistry ; Stomach Neoplasms ; pathology ; U937 Cells ; drug effects

OBJECTIVETo investigate the correlation of the deleted azoospermia (DAZ) gene copy related to gr/gr and b2/b3 deletions in the AZFc region with male spermatogenic impairment.

METHODSThis study included 121 infertile men with different de- grees of spermatogenic impairment and 95 healthy donors from the sperm bank. Using PCR, PCR-RFLP, and Y chromosome specific sequence tagged sites (STS) , we analyzed the association of DAZ gene copy deletions related to gr/gr and b2/b3 deletions in the AZFc region with spermatogenic impairment.

RESULTSThere were 15 cases of gr/gr deletion (12. 40% ) and 6 cases of b2/b3 deletion (4.96%) in the infertility group as compared with 13 cases of gr/gr deletion (13.68%) and 1 case of b2/b3 deletion (1.05%) in the control. Analysis of the DAZ-specific single nucleotide variant (SNV) loci revealed 11 gr/gr-DAZI/DAZ2 deletions (9.09%), 4 gr/gr-DAZ3/DAZ4 deletions (3.31%), and 6 b2/b3-DAZ1/DAZ2 deletions (4.96%) in the infertile men in comparison with 3 gr/ gr-DAZ1/DAZ2 deletions (3.16%), 10 gr/gr-DAZ3/DAZ4 deletions (10.53%), and 1 b2/b3- DAZ3/DAZ4 deletion (1.05%) in the control.

CONCLUSIONPartial deletions of gr/gr and b2/b3 exist in both healthy men and male patients with different degrees of spermatogenic impairment and cannot be considered as a risk factor for spermatogenesis impairment. However, deletions of different DAZ duplicons in gr/gr and b2/b3 deletions have different effects on spermatogenesis. DAZ1/DAZ2 instead of DAZ3/DAZ4 deletions might be associated with spermatogenesis impairment.

Deleted in Azoospermia 1 Protein ; Gene Deletion ; Gene Dosage ; Humans ; Male ; RNA-Binding Proteins ; genetics ; Spermatogenesis ; genetics