Mycoepoxydiene is a novel antitumor agent extracted from marine lignicolous fungi HLY-2, which is Diaporthe phaseolorum by molecule identification. The medium optimization for mycoepoxydiene by orthogonal design and the comparison of submerged fermentation and solid state fermentation were studied. The rusult is that the maximal yield of the compound is 543mg/L, which is 43 times compared to the customary half-seawater PD medium and 15 times to the best submerged condition. This optimum culture medium included potato 250g/L, seawater 300mL/L, glucose 30g/L, lactose 50g/L, KH_ 2 PO_ 4 0.65mmol/L and (NH_ 4 )_ 2 SO_ 4 1g/L in the solid state condition. Differentiation analysis between submerged and solid state fermentation, and antitumor activity of these ferment products were also studied. The antitumor activity of products of the optimum medium approached the pure compound.

Child, Preschool ; Diagnosis, Differential ; Female ; Humans ; Rickets ; diagnosis ; etiology ; therapy ; Treatment Outcome ; Vitamin D Deficiency ; complications

OBJECTIVETo investigate further the possible mechanism of carcinogenesis and portal invasion of hepatocellular carcinoma (HCC).

METHODSSamples of the primary tumors, cancer cells emboli in the portal veins and normal liver tissues adjacent to the tumor were collected from 20 cases of primary HCC. Expression of TIMP-3 (tissue inhibitor of metalloproteinases-3) protein was detected using Western blot. Expression of TIMP-3 mRNA was detected by RT-PCR. Methylation of TIMP-3 gene promoter was detected using methylation-specific PCR (MSP).

RESULTSExpression of TIMP-3 protein and mRNA were obtained in all of the normal liver tissues adjacent to tumor. However, loss of TIMP-3 protein expression was found in 5 and 36 cases respectively in the primary tumors and tumor cell emboli in portal veins. Expression of TIMP-3 protein and mRNA in primary tumors and tumor emboli were significantly lower than that in the normal liver tissues. Promoter methylation of TIMP-3 gene could be detected in primary tumors (7 cases) and cancerous emboli (9 cases) in HCC, while no methylation found in normal liver tissues. In all the HCC cases with promoter gene methylation including primary tumors and cancerous emboli in portal veins, 13 cases showed complete loss and 6 cases showed low expression of TIMP-3 protein and mRNA. Promoter methylation of TIMP-3 was noticed not related with the histological grading of HCC.

CONCLUSIONSThere is a close relationship between loss or low expressions of TIMP-3 and carcinogenesis and portal invasion of HCC. The loss and low expression of TIMP-3 gene and protein were caused by methylation of the gene promoter.

Adult ; Aged ; Blotting, Western ; Carcinoma, Hepatocellular ; chemistry ; genetics ; CpG Islands ; DNA Methylation ; Female ; Humans ; Liver Neoplasms ; chemistry ; genetics ; Male ; Middle Aged ; Promoter Regions, Genetic ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinase-3 ; analysis ; genetics

OBJECTIVETo explore the inhibitory effects of gene expression and functional activity of telomerase in leukemia cell lines by in vitro antisense hTERT treatment.

METHODSAn antisense hTERT eukaryotic expression vector was constructed by using gene recombination technique, targeting the 5' end mRNA sequence of the telomerase catalytic subunit. The vector expression in leukemia cell lines (HL60 and K562) was achieved by transfection using the SuperFect transfection reagent (Qiagen). After transfection, ectopic expression of the telomerase catalytic subunit was analyzed by quantitative fluorescence real-time RT-PCR, and cellular apoptosis and cell cycle parameters were evaluated by flow cytometry respectively.

RESULTSAn antisense pcDNA-hTERT eukaryotic expression vector was successfully constructed. Leukemia cell lines transfected with antisense hTERT constructed displayed a significant inhibition of gene expression of telomerase and its activity in vitro, as compared with the result of the control groups (without transfection and vector control).

CONCLUSIONIn-vitro antisense hTERT expression may down-regulate the gene expression and biological activity of telomerase in leukemia cells, suggesting a possibility of gene therapy against human malignancy through the telomerase-targeted molecular mechanism.

Apoptosis ; Cell Cycle ; DNA-Binding Proteins ; biosynthesis ; genetics ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; HL-60 Cells ; HeLa Cells ; Humans ; K562 Cells ; RNA, Antisense ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Telomerase ; biosynthesis ; genetics ; metabolism ; Transfection

BACKGROUNDRhabdomyosarcoma (RMS) is an uncommon malignancy of the breast. The aim of this study was to summarize its clinicopathologic features and biological behavior.

METHODSFive primary or secondary breast RMSs were collected. Their clinicopathological characteristics and all published literature about breast RMS were reviewed. Immunohistochemical study of desmin, myogenic differentiation 1 (MyoD1), myogenin, leukocyte common antigen (LCA), vimentin, cytokeratin (AE1/AE3), E-cadherin, neuron specific enolase (NSE), CD99, chorioallantoic membrane 5.2 (CAM5.2) and epithelial membrane antigen (EMA) expression were performed.

RESULTSThe five patients were all female with ages ranging from 16 to 46 years old (mean, 30 years). Three were metastatic breast RMSs, two embryonal and one solid variant alveolar, with the primary tumor sites the right labium majus, left nasal meatus and nasopharynx, respectively. The other two, one embryonal and one alveolar, were primaries. Grossly, the surgical specimens revealed round or oval, well-demarcated but nonencapsulated masses. Their cut surfaces consisted of homogeneous grayish yellow or white tissue. Microscopically, most tumor cells were poorly differentiated small round, oval or small polygons with eosinophilic cytoplasm. All cases were positive for vimentin, desmin, MyoD1 and myogenin. One embryonal RMS also had a few cells with perinuclear staining of AE1/AE3. The other markers were negative.

CONCLUSIONSAlthough primary or metastatic RMS in breast was almost confined to young adolescent females, our cases suggested that it can also happen to the middle-aged women. Embryonal RMS has a certain metastatic potential. MyoD1 and myogenin are two useful markers when making differential diagnosis. Axillary lymph node status and age may play a role in the prognosis of primary breast RMS patients.

Adolescent ; Adult ; Breast Neoplasms ; diagnosis ; metabolism ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Rhabdomyosarcoma ; diagnosis ; metabolism ; Young Adult

This study was aimed to investigate the expression of Na(+)/H(+) exchanger 1 (NHE1) in K562 and HL-60 cells undergoing DNA damage induced by etoposide and to elucidate the regulating mechanism. Real-time quantitative PCR (RQ-PCR) and Western blot methods were used to determine the expression of NHE1 in K562 cells after the treating with etoposide. Meanwhile, the flow cytometry was used to detect the apoptosis of leukemic cells. The luciferase reporter vector containing NHE1 promoter was constructed to measure relative luciferase activity after treating with different etoposide concentrations. The results showed that the mRNA and protein of NHE1 increased in accordance with apoptosis ratio in HL-60 cells after treated with etoposide (p < 0.05), but no such obvious increase in K562 cells. Treatment with NHE1 specific inhibitor could block etoposide induced alkalization and reduce the apoptosis ratio of HL-60 cells. The expression pattern and apoptosis alteration was not similar in K562 cells. Relative luciferase activity of reporter vector containing NHE1 promoter however increased in K562 cells after treated with etoposide. It is concluded that the expression of NHE1 is up-regulated in the process of apoptosis of HL-60 cells induced by etoposide and depends on the pHi increasing caused by NHE1 up-regulation which is not found in K562 cells although the transcriptional activity increased.
Apoptosis ; Cation Transport Proteins ; metabolism ; DNA Damage ; Etoposide ; HL-60 Cells ; Humans ; K562 Cells ; Promoter Regions, Genetic ; Sodium-Hydrogen Exchanger 1 ; Sodium-Hydrogen Exchangers ; metabolism

OBJECTIVETo evaluate the variety of non-myeloperoxidase-mediated system activity of neutrophils in newborns during bacterial infection and the effect of cord plasma on the activation of non-myeloperoxidase-mediated system.

METHODSAn infection model with Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) and a non-infection model with phorbol-12-myristate-13-acetate (PMA) were established to investigate the activation of non-myeloperoxidase-mediated system in neutrophils. According to the intensity of fluorescence, the activation of non-myeloperoxidase-mediated system of neutrophils was detected by flow cytometry (FCM). The blood cells and plasma were separated from cord blood and adult blood and cross-mixed in order to investigate the opsonic activity.

RESULTSIn the non-infection model, the activation of non-myeloperoxidase-mediated system with PMA stimulation in cord blood was lower compared with that in adult blood, the statistical difference was significant (t = 3.378, P < 0.01). In the infection model, the activations of non-myeloperoxidase-mediated system in cord blood were also lower compared with those in adult blood, while the statistical difference could only be found in the model with E. coli stimulation (t = 12.150, P < 0.001). Furthermore the experiments demonstrated that cord plasma could deeply depress the non-myeloperoxidase-mediated system activity with E. coli stimulation. On the contrary, adult plasma could successfully recruit the potential of non-myeloperoxidase-mediated system activity of neutrophils in newborns.

CONCLUSIONThe function of neonatal neutrophils might not developed very well. As a stimulant, E. coli failed to induce the non-myeloperoxidase-mediated system activity in neonates, which might be related to the lower level of immunoglobulins in cord blood. This result indicated that immunoglobulins played a more important modulating role in bacterial killing during gram-negative bacterial infections.

Escherichia coli ; immunology ; Fetal Blood ; immunology ; Flow Cytometry ; Humans ; Infant, Newborn ; Neutrophils ; enzymology ; immunology ; Peroxidase ; metabolism ; Staphylococcus aureus ; immunology

OBJECTIVETo study the clinical characteristics of the patients with dengue fever (DF) seen from 2002 to 2006 in Guangzhou in order to prevent and treat dengue fever better.

METHODSClinical data from 1342 inpatients with DF seen from 2002 to 2006 were retrospectively analyzed. The dengue virus was isolated by C6/36 cell culture and genotyped by reverse transcriptase-polymerase chain reaction and gene sequence analysis.

RESULTSThe average age of the patients was 34.4 years, without sex difference in distribution. Most of the patients had obvious toxemic symptoms including fever (100 percent), headache (85.9 percent), myalgia (64.5 percent), bone soreness (46.6 percent) and skin rash (65.9 percent). Leukopenia, thrombocytopenia, elevated alanine aminotransferase, elevated aspartate aminotransferase and hypokalemia were found in 66.0 percent, 61.3 percent, 69.0 percent , 85.7 percent and 28.4 percent of patients, respectively. DF-IgM could be detected in 90 percent of patients. The virus was identified as dengue virus type-I.

CONCLUSIONSThe epidemic of DF was caused by dengue virus- I from 2002 to 2006 in Guangzhou. Most of the patients had classic DF clinical manifestation with high percentage of hepatic injury. Few patients progressed to dengue hemorrhagic fever.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Viral ; blood ; Child ; Child, Preschool ; China ; epidemiology ; Dengue ; diagnosis ; epidemiology ; immunology ; physiopathology ; Dengue Virus ; genetics ; isolation & purification ; Female ; Humans ; Infant ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate the clinical features and prognosis of bone metastases in colorectal cancer patients.

METHODSThe clinical data of 104 cases of colorectal cancer with bone metastasis were collected and retrospectively analyzed.

RESULTSAmong all the 104 patients included, 45 (43.3%) patients had multiple bone metastases, and 59 (56.7%) patients had single bone metastasis. Pelvis (46.1%) was the most common site, followed by thoracic vertebrae (41.3%), lumbar vertebrae (40.4%), sacral vertebrae (29.8%) and ribs (29.8%). One hundred and two patients (98.1%) were complicated with other organ metastases. The median time from colorectal cancer diagnosis to bone metastasis was 16 months, and the median time from bone metastasis to first skeletal-related events (SREs) was 1 month. The most common skeletal-related events (SREs) were the need for radiotherapy (44.2%), severe bone pain (15.4%) and pathologic fracture (9.6%). The median survival time of patients with bone metastases was 10.0 months, and 8.5 months for patients with SREs. ECOG score, systemic chemotherapy and bisphosphonate therapy were prognostic factors by univariate analysis (all P < 0.05). ECOG score and systemic chemotherapy were independent prognostic factors by Cox multivariate analysis.

CONCLUSIONSBone metastasis in colorectal cancer patients has a poor prognosis and the use of chemotherapy and bisphosphonates may have a benefit for their survival.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bone Density Conservation Agents ; therapeutic use ; Bone Neoplasms ; drug therapy ; radiotherapy ; secondary ; Colorectal Neoplasms ; drug therapy ; pathology ; radiotherapy ; surgery ; Diphosphonates ; therapeutic use ; Female ; Follow-Up Studies ; Fractures, Bone ; etiology ; Humans ; Lumbar Vertebrae ; pathology ; Male ; Middle Aged ; Pain ; etiology ; Pelvic Bones ; pathology ; Prognosis ; Retrospective Studies ; Ribs ; pathology ; Sacrum ; pathology ; Spinal Cord Compression ; etiology ; Spinal Neoplasms ; drug therapy ; radiotherapy ; secondary ; Thoracic Vertebrae ; pathology ; Young Adult

Objective To research the role of lymph tracers to protect parathyroid in surgery for papillary thyroid carcinoma.Methods Patients with papillary thyroid carcinoma who met selected criteria were enrolled in this study.Patients were divided into carbon nanoparticle group,methylene blue group,and conventional surgery group.Results No significant complication occurred in the patients of carbon nanoparticle and methylene blue groups.In carbon nanoparticle group,methylene blue group and conventional surgery group,the mean numbers of parathyroid glands detected during surgery were 3.1 ± 0.3,2.9 + 0.4 and 2.3 ± 0.3 (F =3.78,P ＜ 0.01),the rates that parathyroid was cut mistakenly were 1.37％(2/146),2.62％ (2/97) and 7.14％ (6/84) respectively(x2 =17.372,P ＜ 0.05) ; and the incidence of postoperative hypocalcemia were 10.4％ (5/48),9.1％ (3/33) and 17.5％ (7/40,x2 =0.671,P =0.037).Conclusion Thyroid lymphography technique is helpful to protect from the injury to the parathyroid glands in surgery.