OBJECTIVETo investigate the contribution of latent membrane protein (LMP)1 to nasopharyngeal carcinogenesis via Wnt/β-catenin signal pathway.

METHODSThe recombinant plasmid pHA2-LMP1 was constructed; immunofluorescence staining, Dual-Luciferase Reporter Assay, Western blot and immunohistochemistry staining were used to study the effect of LMP1 on the transcriptional activity and expression of β-catenin.

RESULTS(1) Abnormal expression of β-catenin was obtained in 38 cases (50.7%, 38/75), LMP1 expression was obtained in 38 cases (50.7%, 38/75). There was significantly positive correlation between LMP1 expression and abnormal expression of β-catenin in nasopharyngeal carcinoma tissue (P = 0.008). (2) The expression of β-catenin in nuclei of NPC cell line CNE1 and CNE2 transfected with pHA2-LMP1 plasmid dramatically increased, and the expression was remarkable in poorly-differentiated NPC cell line CNE2 than that of well-differentiated CNE1 cells. (3) LMP1 expression dramatically increased the transcriptional activity of β-catenin in CNE1 and CNE2 cells transfected with pHA2-LMP1 and was in a time-dependent. The transcriptional activity of β-catenin was higher in poorly-defferentiated cell line CNE2 than that of well-differentiated NPC cell line CNE1. (4) LMP1 expression did not affect the total protein expression level of β-catenin in both CNE1 and CNE2 cell lines.

CONCLUSIONEB virus-encoded LMP1 may be involved in the pathogenesis of NPC via β-catenin signal pathway.

Adult ; Aged ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Plasmids ; Recombinant Proteins ; metabolism ; Signal Transduction ; Transcriptional Activation ; Transfection ; Viral Matrix Proteins ; metabolism ; Wnt Proteins ; metabolism ; Young Adult ; beta Catenin ; metabolism

OBJECTIVETo improve inversion-polymerase chain reaction (I-PCR) in detection of factor VIII (FVIII) intron 22 inversion for gene diagnosis and prenatal diagnosis of hemophilia A (HA).

METHODSThe modified I-PCR was applied to detect FVIII intron 22 inversion in 8 families with HA. The prenatal diagnosis was performed for 2 pregnant women in the families. The fetal blood samples were obtained by cordocentesis at 22 and 26 weeks gestation respectively.

RESULTSFour of 8 HA families were diagnosed to be FVIII intron 22 inversion. Two fetuses were identified to be normal and one of them was born normal.

CONCLUSIONThe modified I-PCR enables the gene diagnosis and prenatal diagnosis of FVIII intron 22 inversion more accurately and rapidly.

Chromosome Inversion ; Factor VIII ; genetics ; Female ; Hemophilia A ; diagnosis ; genetics ; Humans ; Polymerase Chain Reaction ; methods ; Pregnancy ; Prenatal Diagnosis

This study was undertaken to establish a murine model for unrelated allogeneic umbilical cord blood transplantation (UCBT). The characteristics and percentage of hematopoietic stem/progenitor cells between near-term fetal and neonatal murine peripheral blood (FNPB) and bone marrow (BM) were evaluated by flow cytometry and semisolid methylcellulose culture. BABL/c (H-2(d)) recipient mice conditioned with high dose CTX were transplanted with FNPB form C57BL/6 (H-2(b)) mice and the survival rate, hematopoietic and immunological reconstruction, graft versus host disease (GVHD) and engraftment level were observed. The results showed that the numbers of day 14 CFU-GM and CFU-GEMM in FNPB (176.40 +/- 78.39)% and (141.40 +/- 56.57)%, respectively were much higher than those in BM (75.20 +/- 26.41)% and (68.80 +/- 23.95)%, respectively. Moreover the percentage of Sca-1(+) CD34(+) cell subsets in FNPB (3.63 +/- 1.13)% was also higher than that in BM (1.41 +/- 0.8 7)%. FNPB transplantation improved survival rate and reconstituted hematopoietic and immune function in recipients. There was no evidence of GVHD. Chimeric analysis showed that the proportion of donor cells in BM of recipients was 27.94% at 21 days after transplantation. It was concluded that FNPB contains more hematopoietic stem and progenitor cells with high expansion ability and weak allogeneic immunity, which was similar to human UCB. The murine model for allogeneic UCBT (C57BL/6-->BALB/c) was established successfully.
Animals ; Animals, Newborn ; Fetal Blood ; cytology ; Flow Cytometry ; Graft vs Host Disease ; etiology ; Hematopoietic Stem Cell Transplantation ; Immunity ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Models, Animal ; Transplantation, Homologous

OBJECTIVETo introduce a method of functional reconstruction for total lower lip defects.

METHODSSix patients with lower lip cancer were treated by surgery from November 2007 to February 2009. The full thickness defects with a size from eighty percent to total lower lip were reconstructed with two triangular mental neurovascular island flaps of chin. The musculocutaneous and mucous flaps with neurovascular pedicles were advanced in a V-Y manner to close the defect and reconstruct the orbicularis oris muscle.

RESULTSAll flaps survived and no recurrence and complication were observed and showed excellent cosmetic and functional results over 6 months of follow-up. The reconstructed lower lips provided a wide-enough mouth opening, and the sensation of oral mucosa and the muscle function of oral sphincter were preserved.

CONCLUSIONSThe technique of mental neurovascular V-Y advancement island flap can get satisfactory results in both functional and the aesthetic perspects.

Aged ; Carcinoma, Squamous Cell ; surgery ; Carcinoma, Verrucous ; surgery ; Female ; Follow-Up Studies ; Humans ; Lip ; injuries ; surgery ; Lip Neoplasms ; surgery ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps

OBJECTIVETo obtain recombinant fusion protein HSA (human serum albumin)-PTH(1-34) in Pichia pastoris.

METHODSHSA and PTH(1-34) cDNA were obtained with PCR and the DNA segments were cloned into vector pPIC9 with linker. The linearized plasmids were transformed GS115 competent cells treated with LiCl, and mut+ transformants were screened on MD plate. With AOX promoter and alpha-MF signal sequences leading, fusion protein was expressed in GS115. PCR and SDS-PAGE were employed to confirm the integration and expression of HSA-PTH(1-34). The fusion protein was identified by Western blotting and classical adenylate cyclase assay.

RESULTThe PCR results showed that the gene of HSA-PTH(1-34) was integrated into GS115 genome. Western bolt approved the existence of two domains of HSA and PTH(1-34). The bioactivity assay in rabbit cortical membranes indicated that HSA-PTH (1-34) activated adenylate cyclase, but the activity was lower than that of the synthetic PTH(1-34).

CONCLUSIONActive fusion protein HSA-PTH (1-34) is successfully expressed in Pichia pastoris.

Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Peptide Fragments ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Serum Albumin ; biosynthesis ; genetics ; Teriparatide ; analogs & derivatives

OBJECTIVETo observe the effect of compound gengniankang (GNK) in regulating the endocrine and immune functions in aged female rats.

METHODSAged female rats with osteoporosis were selected as the object for observation and healthy young rats were taken for control. Animals were administered by GNK and nilestriol respectively, through gastric perfusion, for 3 months to observe the therapeutic effect of drug treatment on osteoporosis and the regulatory effect on endocrine and immune function. Bone mineral density (BMD) was measured by double energy X-ray absorption technique, serum levels of estradiol (E2), follicule-stimulating hormone (FSH) and luteinizing hormone (LH) were determined by RIA, T-cell subsets and apoptosis in spleen were detected by flow cytometry.

RESULTSIn aged rats with osteoporosis, the BMD decreased, serum level of E2 lowered, FSH and LH levels raised, splenic CD4+, CD4+/CD8+ significantly decreased and T-cell apoptosis rate significantly elevated. GNK could increase the BMD, lower the FSH and LH levels, but showed no significant effect on E2 level. It could increase the CD4+ and CD4+/CD8+ ratio to nearby the normal range, and reduce the apoptosis of T-cells.

CONCLUSIONGNK has therapeutic effect on osteoporosis in aged rats, and is able to regulate the endocrine and enhance the immune function in organism.

Absorptiometry, Photon ; Aging ; Animals ; Apoptosis ; drug effects ; Bone Density ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Estradiol ; blood ; Female ; Follicle Stimulating Hormone ; blood ; Luteinizing Hormone ; blood ; Osteoporosis ; blood ; immunology ; Rats ; Rats, Sprague-Dawley ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes ; pathology

OBJECTIVETo compare the therapeutic effect of Compound Recipe Gengniankang ( GNK) with that of hormone replacement treatment (HRT) on climacteric female rats with osteoporosis, and to investigate the roles of estrogen and estrogen receptors in the mechanism of osteoporosis.

METHODSClimacteric female rats with osteoporosis were chosen and divided into three groups (GNK group, HRT group and control group). Apoptosis of ovarian granulose cells was measured by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) assay. Serum level of estradiol (E(2)), follicle stimulating hormone (FSH), luteinizing hormone (LH) were determined by the method of radioimmunoassay (RIA). Reverse transcriptase polymerase chain reaction (RT-PCT) technology was used to evaluate the expression of estrogen receptor (ER) in bone. Bone mineral density (BMD) was measured by double energy X-ray absorption (DEXA).

RESULTSIn the climacteric rats, BMD, serum E(2), ER mRNA expression in bone decreased remarkably, and serum FSH, LH and apoptosis of ovarian granulose cells increased obviously. After treating with GNK, all the indexes were reversed except serum E(2). The increase of E(2) was not significant.

CONCLUSIONGNK is effective on climacteric osteoporosis female rats. Its role is performed not by increasing serum E(2) but by enhancing ER in the bone and inhibiting apoptosis of ovarian granulose cells. GNK can deter further exhaustion of ovarian function.

Age Factors ; Animals ; Apoptosis ; Bone Density ; drug effects ; Bone and Bones ; drug effects ; metabolism ; Climacteric ; Drugs, Chinese Herbal ; pharmacology ; Estradiol ; blood ; Female ; Follicle Stimulating Hormone ; blood ; Hormone Replacement Therapy ; Hormones ; blood ; Luteinizing Hormone ; blood ; Osteoporosis ; metabolism ; Ovary ; drug effects ; physiology ; Rats ; Receptors, Estrogen ; biosynthesis

OBJECTIVEThe development status of pulmonary artery is one of the most important criteria for decision-making strategy and predicting postoperative outcome in congenital heart disease with decreased pulmonary blood flow. Currently, Nakata index and McGoon index have been used as morphologic index in evaluating the development status of pulmonary artery. Those indices have some shortcoming. It was recently found that pulmonary veins index is a more precise morphological indicator of pulmonary blood flow and development status of pulmonary vessels. This study aimed to explore an index of evaluating pulmonary blood stream and the development of pulmonary vessels, as a criterion for surgical decision-making strategy.

METHODSThe diameters of left and right pulmonary arteries and pulmonary veins were measured on DSA films in 74 patients with congenital heart disease with decreased pulmonary blood flow, The correlative analysis was done between Nakata index, McGoon index, pulmonary vein index (PVI) and postoperative outcome which were the length of stay in ICU, duration of mechanical ventilation and dose of inotropic drugs.

RESULTSExcellent correlations between the size of pulmonary veins and pulmonary arteries were found, the correlation between left pulmonary veins and distal portion of left pulmonary artery was 0.73, between left pulmonary veins and proximal portion of left pulmonary artery was 0.72, right pulmonary veins and distal portion of right pulmonary artery was 0.67, and right pulmonary veins and proximal portion of right pulmonary artery was 0.71. The length of stay in ICU, duration of mechanical ventilation and dose of inotropic drugs correlated well with PVI (r = -0.51, -0.478, and -0.693). Compared with Nakata index and McGoon index, PVI was a better criterion for evaluating the developmental status of the whole pulmonary vessels. In the right ventricular outlet reconstruction patients, the McGoon index for patients with low cardiac output syndrome (LCOS) was 1.36 +/- 0.51, and 1.97 +/- 0.58 for patients without LCOS (t = 2.347, P < 0.05), the Nakata index for patients with LCOS was 164 +/- 106 mm(2)/m(2) and 269 +/- 124 mm(2)/m(2) for patients without LCOS (t = 2.218, P < 0.05), the PVI for patients with LCOS was 152 +/- 77 mm(2)/m(2) and 273 +/- 125 mm(2)/m(2) for patients without LCOS (t = 2.936, P < 0.01), pulmonary vessel index of patients with LCOS was less than that of those without LCOS. When PVI was < or = 180 mm(2)/m(2), postoperative hemodynamics was unstable, the frequency of low cardiac output syndrome and mortality significantly increased.

CONCLUSIONSThe development of pulmonary arteries and pulmonary veins correlated with each other. PVI is a precise morphological indicator of pulmonary blood flow and development of pulmonary vessels. It is a helpful indicator to decide surgical strategy.

Adolescent ; Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; physiopathology ; Hemodynamics ; Humans ; Infant ; Male ; Pulmonary Artery ; growth & development ; physiopathology ; Pulmonary Veins ; growth & development ; physiopathology

Needle sticking method, which can be combined with multiple needling techniques, has been attached with great importance in recent years by doctors in clinic. Combining with the clinical experiences, the authors expounded the needle sticking method through its unified concept, differences between needle sticking method, which was an acupuncture technique, and stuck needle which was an accident during acupuncture, selection of needles, manipulations, mechanism of treatment, range of application, attentions and advantages of popularization. It is held that the technique can be widely applied for treatment of acute and chronic diseases of various departments with filiform needles. Easy to be manipulated, understood and mastered, the technique is without side effect and valuable to be popularized.
Acupuncture Therapy ; instrumentation ; methods ; Humans ; Medicine, Chinese Traditional

OBJECTIVETo study the effects of cadmium chloride in anterior pituitary and the relation between apoptosis and expression of procaspase-9 mRNA.

METHODSIn vivo studies:40 SD male rats were randomly distributed into four groups which were administered with CdCl2 at different doses by gavage for 6 weeks;

IN VITRO STUDIESthe rats' anterior pituitary cells were primarily cultured for 120 hours, then treated with CdCl2 at the dose of 0, 1.56, 3.12, 6.25, 12.50, 25.00, 50.00, 100.00 micromol/L for 6 hours; The indices included: expression of procaspase-9 mRNA, detection of apoptosis with TUNEL assay.

RESULTSThe results showed the excretion of ACTH, LH seemed to be decreased dramatically and the apoptosis inclined to enhance remarkably, and further more, the expression of procaspase-9 mRNA appeared to be increased significantly as compared with those of the control. It was show that a dose-effect relationship between the CdCl2 dosing and indices above with the regression analysis and a linear correlation between the mean gray value of apoptosis cell and the relative gray value of procaspase-9 mRNA positive cell. The results indicated that damnification, for example, apoptosis could be caused by certain dose of CdCl2 in anterior pituitary cells with dose dependent manner. Caspase-9 might play a role in the occurrence of apoptosis.

CONCLUSIONIt was suggested that cadmium could induce apoptosis of anterior pituitary both in vivo and in vitro in the manner of dose-dependent, and caspase-9 might play a role during above processes.

Animals ; Apoptosis ; drug effects ; Cadmium Chloride ; administration & dosage ; toxicity ; Caspase 9 ; genetics ; Cells, Cultured ; Dose-Response Relationship, Drug ; Male ; Pituitary Gland, Anterior ; cytology ; drug effects ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects