Objective To evaluate the feasibility and effectiveness of combination chemotherapy with etoposide and cisplatin(EP)regimen on the patients with high-risk,chemorefractory and recurrent gestational trophoblastic neoplasia(GTN).Methods Thirty-nine patients with gestational trophoblastic tumors were analyzed retrospectively,25 of 39 patients were of high-risk,9 patients were chemorefractory and 5 patients were recurrent.All 39 patients were administrated with EP regimen,and 10 patients were assisted with surgery.All the patients were followed up.Clinical response,toxicity,the occurrence of secondary tumors of all patients,and the fertility of 30 patients whose fertility function was preserved were investigated. Results Thirty-nine GTN patients underwent a total of 221 cycles of the EP regimen.The average number of courses for each patient was 5.7.The total complete remission rate of the regimen was 74%(29/39). Twenty-five patients with high-risk GTN received a total of 139 cycles and the average number of courses was 5.6.Nineteen patients achieved complete remission and 6 patients showed drug-resistant.The complete remission rate of the high-risk group was 76%(19/25).Nine patients with chemorefractory GTN obtained a total of 55 cycles and the average number of courses was 6.1.Six patients achieved complete remission and 3 patients showed drug-resistant again.The complete remission rate of the chemorefractory group was 6/9. Five patients with recurrent GTN received 27 cycles and the average number of courses was 5.4.Four patients achieved complete remission,1 patient showed drug-resistance and died.Bone marrow toxicity, gastrointestinal reaction and alopecia were the main side effects of the EP regimen,but the bone marrow toxicity was slight and no grade Ⅳ side effect occurred.No fatal effect was found.Eight of 30 patients whose fertility faction was preserved had become pregnant after recovery,with a total of 8 pregnancies.Among them,2 were terminated by induced abortion,and 6 underwent normal term delivery and gained 6 infants who had no congenital malformation.All the 6 children had normal growth and development after childbirth. None of the women developed secondary tumors.Conclusion The EP regimen is effective and safe for the treatment of high-risk,chemorefractory and recurrent GTN.

BACKGROUND: Endothelial progenitor calls are the cells that can form new blood vessels in the way of angiogenesis in the body,which updates the conventional theory of angiogenesis, vascular damage and repair after birth and provides new ideas for research and treatment of ischemic diseases.OBJECTIVE: To investigate the effects of dog umbilical cord blood endothelial progenitor call (UCB-EPC) transplantation on angiogenesis after myocardial infarction.DESIGN, TIME AND SETTING: An in vivo cytological experiment was performed at the Laboratory Center of Xinhua Hospital between May 2006 and March 2007.MATERIALS: One full-term pregnant hybrid dog was included for preparation of UCB-EPCs. Thirty-six adult dogs were randomly divided into a cell transplantation group (n = 18) and a model control group (n = 18).METHODS: Acute myocardial infarction model was established in each group by ligation of antedor descending coronary artery.In the cell transplantation group, 2 mL physiological saline containing 5×10<'6> BrdU-labeled EPCs was injected into the coronary artery, while in the model control group, simple physiological saline of the same amount was given. At 1,4, and 8 weeks after transplantation, dogs were sacrificed for harvesting myocardial tissue.MAIN OUTCOME MEASURES: Myocardial infarction was confirmed by hematoxylin-eosin staining. Myocardial angiogenesis was observed by BrdU immunohistochemical staining. The number of infarcted myocardial vessels was calculated by yon Willebrand (vW) factor staining.RESULTS: There was plenty of scar tissue, flbroblasts, and small vessels in the myocardial infarction region. In the cell transplantation group, brown yellow particles (BrdU-positive expression) appeared in some nuclei in small vessels from infarcted myocardium. Newly formed vessels were not found in the model control group. In the cell transplantation group, brown yellow particles (vW factor-positive expression) appeared in the cytoplasm of the vascular endothelial cells in the myocardial ischemia and infarction regions, vW factors were not expressed in the model control group. At 1, 4, and 8 weeks after myocardial infarction,there was no significant difference in vessel counts no matter in myocardial ischemia region or in myocardial infarction region between the call transplantation and model control groups.CONCLUSION: EPCs derived from UCB of pregnant dog can participate in the formation of blood vessels but can not promote angiogenesis after acute myocardial infarction.

Ambroxol and clenbuterol were extracted from human plasma samples by liquid-liquid extraction, ambroxol was separated on a Zorbax XDB-C18 column and detected by tandem mass spectrometry with an atmospheric pressure chemical ionization interface after oral administration of a compound preparation. Clenbuterol was separated on a Zorbax XDB-C8 column and detected by tandem mass spectrometry with an electrospray ionization interface. Diphenhydramine is used as the internal standard. The linear concentration ranges of the calibration curves for ambroxol and clenbuterol were 0.080 - 400 microg x L(-1) and 5.0 - 5 000 ng x L(-1), respectively. The lower limits of quantification were 0.080 microg x L(-1) for ambroxol and 5.0 ng x L(-1) for clenbuterol, individually. The inter-day and intra-day precision (RSD) across three validation run over the entire concentration range was below 7.5%, and the accuracy (RE) was within +/- 2.5% for both ambroxol and clenbuterol. The methods were used to determine the pharmacokinetic parameters of ambroxol and clenbuterol in human plasma after oral administration of a compound preparation containing 60 mg ambroxol hydrochloride and 40 microg clenbuterol hydrochloride. The method was proved to be highly sensitive, selective and suitable for the pharmacokinetic study of different compound preparations containing ambroxol and clenbuterol.
Administration, Oral ; Adrenergic beta-Agonists ; administration & dosage ; blood ; pharmacokinetics ; Adult ; Ambroxol ; administration & dosage ; blood ; pharmacokinetics ; Area Under Curve ; Chromatography, Liquid ; methods ; Clenbuterol ; administration & dosage ; blood ; pharmacokinetics ; Diphenhydramine ; standards ; Expectorants ; administration & dosage ; analysis ; pharmacokinetics ; Humans ; Male ; Reference Standards ; Reproducibility of Results ; Tandem Mass Spectrometry ; methods

Optimization of cultivation condition of recombinant E. coli DH5 alpha/pDH-B2m and the condition suitable for expression of recombinant mature peptide of human bone morphogenetic protein-2 was carried out in 500 mL shaking flasks and then transferred to NBS Bioflo IV, a 20 L DO feed-back fed-batch culture system, to obtain rhBMP-2. The results indicate that keeping dissolved oxygen at 40% and controlling nutrient feeding rate with DO feed back strategy can obtain theoretically 3.59 g recombinant mature peptide of hBMP-2 per liter of broth, the final cell density OD600 reaches 57(22.8 g dry cell weight/L), and the expression of rhBMP-2 amounts to 30% of the total protein in E. coli.
Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; biosynthesis ; genetics ; Culture Media ; Escherichia coli ; genetics ; Fermentation ; Glycerol ; pharmacology ; Recombinant Proteins ; biosynthesis ; Time Factors ; Transforming Growth Factor beta

An esophageal squamous cell carcinoma measuring 18.3 cm in length and 5 cm in diameter was found in the mediastinum of a 53-year man. The patient underwent a modified 3-stage esophagectomy and an esophagogastrostomy at the cervical level (Wu's method). The operation was performed smoothly and the patient recovered uneventfully after the operation. The patient was followed up for 6 months after discharge and reported no difficulties in eating with improved quality of life. This case represents the world's longest esophageal cancer that had been surgically removed. Local advanced esophageal cancer should be removed immediately to prevent potential occurrence of esophageal obstruction, tracheoesophageal fistula or aorto-esophageal fistula.
Carcinoma, Squamous Cell ; surgery ; Esophageal Fistula ; Esophageal Neoplasms ; surgery ; Esophageal Stenosis ; Esophagectomy ; Female ; Humans ; Male ; Middle Aged ; Quality of Life

OBJECTIVETo explore the inhibitory effect of migration-inducing gene 7 (Mig-7) gene silencing induced by retroviral-mediated small hairpin RNA (shRNA) on vasculogenic mimicry (VM), invasion and metastasis of human hepatocellular carcinoma (HCC) cells in vitro.

METHODSTwo target sequences (Mig-7 shRNA-1 and Mig-7 shRNA-2) and one negative control sequence (Mig-7 shRNA-N) were synthesized. The recombinant retroviral vectors carrying Mig-7 shRNA were constructed, and HCC cell line MHCC-97H were transfected with Mig-7 shRNA-1, Mig-7 shRNA-2, Mig-7 shRNA-N, or the empty vector, or treated with 125 µg/mL recombinant human endostatin (ES). Mig-7 expression in the treated cells was detected using semi-quantitative PCR and Western blotting. The inhibitory effect of Mig-7 silencing on VM formation was investigated in a 3-dimensional cell culture system; the changes in cell adhesion, invasion and migration were assessed with intercellular adhesion assay, Transwell invasion assay and Transwell migration assay, respectively.

RESULTSThe expression of Mig-7 at both mRNA and protein levels decreased significantly, VM formation, invasion and metastasis were suppressed, while intercellular adhesion increased significantly in MHCC-97H cells in Mig-7 shRNA-1 and Mig-7 shRNA-2 groups (P<0.05); such changes were not observed in cells transfected with Mig-7 shRNA-N or the empty vector, nor in cells treated with ES.

CONCLUSIONSMig-7 silencing by retroviral-mediated shRNA significantly inhibits VM formation, invasion and metastasis and increases the intercellular adhesion of the HCC cells, while ES does not have such inhibitory effects.

BACKGROUNDCarbon dioxide (CO2) laser soldering is an alternative technique for tissue bonding. Basic fibroblast growth factor (bFGF) and transforming growth factor β(1) (TGFβ(1)) are two key factors for wound healing. This study was performed to demonstrate the efficacy of CO2 laser soldering for dural reconstruction and the effect of bFGF and TGFβ(1) on healing.

METHODSIn Part I, 10 minipigs were randomized into two equal groups. Dural defects were reconstructed by conventional fibrin glue bonding (group I(a)) or CO2 laser soldering (group I(b)). The reconstructed dura was subjected to burst pressure (BP) measurement and immunohistochemical staining after 1 week. In Part II, 36 minipigs were randomized into three equal groups. Dural reconstruction was achieved by CO2 laser soldering. Exogenous bFGF (group II(b)) or TGFβ(1) (group II(c)) was administered while group II(a) served as a control group. The specimens were subjected to BP measurement after 1, 2, 3, and 4 weeks, respectively.

RESULTSIn Part I, the dura specimens displayed positive staining of only bFGF in group I(a) and of both bFGF and TGFβ(1) in group I(b). Group I(b) showed higher BP than group I(a) ((98.00 ± 21.41) mmHg vs. (70.80 ± 15.09) mmHg, respectively; P < 0.05). In Part II, BP of group II(c) was significantly higher than that of group II(a) (P < 0.01). The BP of group II(a) trended toward stabilization after 3 weeks of growth, while that of groups II(b) and II(c) trended toward stabilization after 2 weeks of growth.

CONCLUSIONSCO2 laser soldering is a reliable technique for dural reconstruction. The superior healing of dural reconstruction by CO2 laser soldering may be related to higher expression of bFGF and TGFβ(1), and CO2 lasers may stimulate their secretion. Exogenous bFGF or TGFβ(1) may improve healing by shortening the wound healing time, and exogenous TGFβ(1) may improve the tensile strength.

Animals ; Dura Mater ; drug effects ; surgery ; Female ; Fibrin Tissue Adhesive ; chemistry ; Fibroblast Growth Factor 2 ; therapeutic use ; Immunohistochemistry ; Lasers, Gas ; Male ; Swine ; Swine, Miniature ; Transforming Growth Factor beta1 ; therapeutic use ; Wound Healing ; drug effects

OBJECTIVETo study the biological properties of human dental pulp cells (HDPC) by cloning and analysis of genes differentially expressed in HDPC in comparison with human gingival fibroblasts (HGF).

METHODSHDPC and HGF were cultured and identified by immunocytochemistry. HPDC and HGF subtractive cDNA library was established by PCR-based modified subtractive hybridization, genes differentially expressed by HPDC were cloned, sequenced and compared to find homogeneous sequence in GenBank by BLAST.

RESULTSCloning and sequencing analysis indicate 12 genes differentially expressed were obtained, in which two were unknown genes. Among the 10 known genes, 4 were related to signal transduction, 2 were related to trans-membrane transportation (both cell membrane and nuclear membrane), and 2 were related to RNA splicing mechanisms.

CONCLUSIONThe biological properties of HPDC are determined by the differential expression of some genes and the growth and differentiation of HPDC are associated to the dynamic protein synthesis and secretion activities of the cell.

Cloning, Molecular ; Cloning, Organism ; Dental Pulp ; Fibroblasts ; Gene Library ; Gingiva ; Humans ; Polymerase Chain Reaction

OBJECTIVETo review the clinical features and therapeutic experience in children with plastic bronchitis.

METHODSFourteen children with plastic bronchitis were reviewed retrospectively, 12 of which were under two years old. The clinical features are characterized by sudden onset, episodes of profound hypoxia and respiratory tract obstruction. SaO2 was between 0.70 and 0.80 even with mask oxygen inhalation. Eight cases were pyretic, 4 cases expectorated jel-like bronchial casts. The chest X-ray picture showed patchy consolidation or atelectasis unilaterally (10 cases) or bilaterally (2 cases). Pulmonary marking thickening and patchy shadow were observed in 2 cases. Twelve cases underwent rigid bronchoscopy and the bronchial casts were removed. Two cases underwent endotracheal intubation.

RESULTSEight cases of 12 children received therapeutic bronchoscopy were cured. Other 4 cases had second therapeutic bronchoscopy and bronchial casts were removed again in 3 cases, one died from pulmonary hemorrhage. Two cases who underwent endotracheal intubation died from the multiple organ failure (MOF). Pathologic results showed:the bronchial casts were composed mainly of mucus and fibrin, inflammatory cell infiltrate were observed in 6 cases (Type 1, inflammatory), no cellular infiltrate occurred in 8 cases (Type 2, acellular).

CONCLUSIONSPlastic bronchitis is a severe and dangerous disease. The branching plastic casts may obstruct part or the entire tracheobronchial, causing respiratory failure. Bronchoscopy and pathologic examination are essential for it's diagnosis and treatment.

Airway Obstruction ; Bronchitis ; etiology ; pathology ; surgery ; Bronchoscopy ; Child ; Child, Preschool ; Female ; Humans ; Hypoxia ; Infant ; Male ; Pulmonary Atelectasis ; Retrospective Studies

OBJECTIVETo study the role of extracellular signal-regulated protein kinase 5 (ERK5) during the biosynthesis of follicle-stimulating hormone (FSH)-mediated progesterone in primary granulosa cells.

METHODSThe expressions of phosphorylated and general forms of ERKS in primary granulosa cells after the treatment of FSH were detected by Western blot analysis. The subcellular localization of ERK5 was observed by confocal microscopy. The effect of ERK5 on FSH-mediated progesterone biosynthesis in primary granulosa cells was analyzed using recombinant adenovirus vectors.

RESULTSERK5 activation was induced by FSH in a time-dependent manner in primary cultured granulosa cells, although the general ERK5 protein level decreased also in a time-dependent manner. The treatment of FSH showed no remarkable effect on the subcellular distribution of endogenous ERK5, which was mainly in the cytoplasm of granulosa cells. The co-infection of Ad-caMEK5 and Ad-wtERK5 increased the progesterone production and StAR expression in primary cultured granulosa cells, whereas inhibition of ERK5 activation suppressed the FSH-stimulated progesterone production.

CONCLUSIONERK5 may stimulate FSH-mediated progesterone production in primary cultured granulosa cells.

Animals ; Cells, Cultured ; Female ; Follicle Stimulating Hormone ; pharmacology ; Granulosa Cells ; drug effects ; metabolism ; Mitogen-Activated Protein Kinase 7 ; metabolism ; physiology ; Progesterone ; biosynthesis ; Rats ; Rats, Sprague-Dawley