Objective To investigate the effect of silk fibroin scaffolds seeded with adipose mesenchymal stem cells (ADMSCs) in remodeling fiber for tunica albuginea defect in rabbits.Methods Fifty-six New Zealand rabbits were divided into 4 groups according to the random number table after a defect was created in the tunica albuginea:Group A (the defect was repaired with silk fibroin),Group B (with silk fibroin seeded with ADMSCs),Group C (with autologous tunica vaginalis) and Group D (left unrepaired),with 14 rabbits per group.Tunica albuginea sections were obtained for HE staining,Sirius red staining,Hart staining and immunofluorescence staining of macrophages at 6 and 12 weeks after surgery.Results At 12 weeks after surgery,HE staining revealed chaotically distributed new fiber ingrowth in Group D and orderly ingrowth in Groups A,B,and C.At 12 weeks after surgery,Sirius red staining revealed mean area of type Ⅰ collagen fibers was greater than that of type Ⅲ in Groups A (98 780 ±4 190 vs 51 177 ±5 464),B(94 855 ±9 010 vs 50 815 ±3 895),and C(99 860 ±6 015 vs 50 948 ± 6 595),but the difference in area of collagen fiber of the same type was insignificant among the three Groups.Moreover,less type Ⅰ collagen fibers (79 386 ±2 237) and more type Ⅲ collagen fibers (85 278 ± 2 645) were observed in Group D compared with other three Groups (P ＜ 0.01).At 12 weeks after surgery,Hart staining showed the mean area of elastic fibers in Groups A,B,C,and D was 2 805 ± 90,2 849 ±84,3 068 ±485,and 1 961 ±96 respectively.There was no statistical difference between Groups B and C,but less amount of fibers was observed in Group A (P ＜ 0.01) and least amount was observed in Group D (P ＜0.01).At 6 weeks after surgery,the number of infiltrating macrophages in Groups A,B,C and D was 4.10 ± 0.87,3.80 ± 0.78,3.70 ± 0.94,and 6.80 ± 1.63 respectively.There was no statistical difference in the number of macrophage infiltration among Groups A,B and C,but all were lower than that in Group D (P ＜ 0.01).Conclusion Silk fibroin seeded with ADMSCs is comparable to autologous grafts for repair of tunica albuginea defect in rabbits.

Objective:To improve the genetic stability of HBV gene in transgenic mice.Methods:HBV transgenic mice were bred by backcross and double cross.The HBV gene expression and replication were studied with real-time PCR,ELISA and chemiluminescence.Results:The HBV transgenic mice have stably bred to 23rd generation.The serum HBsAg level is 4122.31?2044.74IU/ml;The rate of HBV transgenic mice whose serum HBV DNA reach 104~106copies/ml was 93.93%.The HBV replication and expression were improved markedly.There is no difference between male and female mice about serum HBsAg level.Conclusion:After breeding the HBV gene was expressed stably with high-level in transgenic mice.

OBJECTIVETo investigate whether the fetal immune tolerance induction could replace the HLA typing for hematopoietic stem cell transplantation.

METHODSImmune tolerance of SD rats was induced by injecting host Wistar rats peripheral blood mononuclear cells into yolk sac of the embryo, afterward the mature male offsprings were used as donor. The host female recipients received lethal dose irradiation and bone marrow transplantation(BMT). The Wistar rats transplanted with bone marrow from donor and unrelated SD rats as well as the rats which received radiation alone were used as control. The survival, histopathologically GVHD, the mental status, food and water intake, coat characteristics, activities were observed. Forty days after BMT, autologous and allogenous skin transplantation between donor and recipient rats was performed to observe the engraftment of solid organ.

RESULTSThe survival of the rats received bone marrow grafts from the immune tolerant donor was significantly longer than that of control groups (30 day survival rates were 86.7%, 6.7%, 0%, and 0% respectively), and there was no histopathologically GVHD observed, while in the sham group, the manifestations of GVHD was clearly visible. The skin engraftment rate between the host and the immune tolerant donor was significantly higher than that among non-related rats (84.6% and 0% respectively).

CONCLUSIONThe induction of immune tolerance in embryo can overcome the HLA barrier and provide a good donor for hematopoietic stem cell and solid organ transplantation.

Animals ; Embryo, Mammalian ; immunology ; Female ; Graft vs Host Disease ; immunology ; prevention & control ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Testing ; Immunosuppression ; Male ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Transplantation Chimera

This study was aimed to investigate the clinical value of detecting BCR/ABL fusion gene by fluorescence in situ hybridization (FISH). The conventional cytogenetic test and detection of BCR/ABL fusion gene by FISH for bone marrow of patients with newly diagnosed chronic myeloproliferative disease or myelodysplastic and myeloproliferative disorders, acute lymphocytic leukemia and chronic myelogenous leukemia (CML) after allogeneic hematopoietic stem cell transplantation were carried out. The results showed that (1) out of 46 newly diagnosed as chronic myeloproliferative disease or myelodysplastic and myeloproliferative disorders, 22 cases were diagnosed as CML, the FISH detection showed all positive (100%), while cytogenetic test showed 86.4% (19/22) positive, in the other 24 patients who were diagnosed as other chronic myeloproliferative disease or myelodysplastic and myeloproliferative disorders, BCR/ABL fusion gene all were be detected as negative 100% by FISH, while the cytogenetic test of bone marrow in 3 cases supported the diagnosis of CML, and the diagnosis of myelodysplastic disorder in 1 case; (2) in 3 out of 7 acute lymphocytic leukemia cases the BCR/ABL fusion gene could not be detected by FISH; (3) the BCR/ABL fusion gene could be detected by FISH in 2 cases of CML received allogeneic hematopoietic stem cell transplantation, with abnormal threshold 6.5% and 1.2% respectively. It is concluded that the detection of BCR/ABL fusion gene by FISH is sensitive and reliable, which is very important for the diagnosis and differential diagnosis of chronic myeloproliferative disorders, myelodysplastic and myeloproliferative disease, as well as definite diagnosis of Ph(+) acute lymphoblastic leukemia. This method also has an important significance for monitor of minimal residual disease in CML patients received allogeneic hematopoietic stem cell transplantation.
Adolescent ; Adult ; Aged ; Child ; Female ; Fusion Proteins, bcr-abl ; genetics ; Genes, abl ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Leukemia ; diagnosis ; genetics ; Male ; Middle Aged ; Myeloproliferative Disorders ; diagnosis ; genetics ; Young Adult

Objective To study the effect of artesunate on inflammatory responses to severe pneumonia by regulating macrophage migration inhibitory factor (MIF) in rats.Methods Total of 100 SD by random (random number) assigned,20 rats were control group,80 SD rats with severe pneumonia were caused by Klebsiella pneumoniae,60 SD rats were treated with different concentrations (20,40,80 mk/kg) of artesunate after modeling.The pathological changes of lung tissue,the level of MIF myeloperoxidase activity and inflammatory cell infiltration in lung tissue of rats were evaluated.Results After treatment with artesunate,the severity of inflammation was significantly alleviated in rats with severe pneumonia evidenced by decrease in myeloperoxidase activity [severe pneumonia:(17.5 ± 1.5) vs.treatment group:(7.5 ±2.0)] and reduction in inflammatory cell infiltration (severe pneumonia:27 × 106 vs.treatment group:12.5 × 106).Similarly,the artesunate also reduced the production of inflammatory cytokines significantly in bronchoalveolar lavage fluid (IL-1 in severe pneumonia group:(1 100 ± 50) pg/ml vs.treatment group:(400 ± 60) pg/ml;IL-6 in severe pneumonia group:(700-± 30) pg/ml vs.treatment group:(200 ±40) pg/ml;IL-10 in severe pneumonia group:(500 ± 70) pg/ml vs.treatment group:(200 ± 40) pg/ml;TNF-αin severe pneumonia group:(500 ± 80) pg/ml vs.treatment group:(150 ± 50) pg/ml.In addition,artesunate inhibited the level and production of MIF,thus inhibiting the inflammatory responses mediated by MIF.Conclusions Artesunate had a protective effect on pneumonia caused by Klebsiella pneumoniae in rats via inhibiting the inflammation responses mediated by MIF.This study provided a molecular basis for newly developed drugs applied to the treatment of pneumonia caused by Klebsiella pneumoniae in rats.

OBJECTIVETo evaluate the perioperative risk factors of the cleft palate repair in Pierre Robin sequence patients at early age and to investigate how to control the risk factors.

METHODSSix consecutive patients with Pierre Robin sequence underwent primary repair of cleft palate in Department of Oral Maxillofacial Surgery, Peking University School of Stomatology from June 2001 to February 2004. The patients underwent von Longenbeck operation by the same perioperative observation of serum oxygen saturation were obtained for these patients. patients included 4 males and 2 females with age of 9 months to 5 surgeon. Pre- and post-operative polysomnographic studies and years.

RESULTSAll the patients suffered various degree of hypoxaemia during the period of intubation. There was only one patient who had hypoxaemia within the first 2 hours during postanaesthetic recovery period. No obvious difference was found in apnea and hypopnea index (AHI) among the patients before and after operation.

CONCLUSIONSSevere hypoxaemia may happen in perioperative period when the patients with PRS underwent cleft palate repair. Most patients with PRS could undergo cleft palate repair safely performed by experienced surgeon at early age under comprehensive consideration and careful control of the risk factors.

Child, Preschool ; Cleft Palate ; surgery ; Female ; Humans ; Hypoxia ; etiology ; Infant ; Intraoperative Complications ; therapy ; Male ; Pierre Robin Syndrome ; surgery ; Postoperative Complications ; therapy ; Risk Factors

OBJECTIVETo investigate the diversify of the nuclear pathway of c-Jun NH2-terminal kinases (JNK) during transient brain ischemia/reperfusion injury in hippocampal neuron apoptosis in spontaneously hypertensive rats (SHR) and to test whether the neuroprotection of curcumine on transient brain ischemia/reperfusion injury in SHR is related to the nuclear pathway of JNK.

METHODSMale Wistar-Kyoto (WKY) rats and SHR were randomly divided into five groups (n = 6): WKY sham group (W-Sham), WKY ischemia/reperfusion group (W-I/ R), SHR sham group (S-Sham), SHR ischemia/reperfusion group (S-I/R) and SHR curcumine (a chinese traditional medicine)100 mg/kg treatment group (S-Cur), which were sacrificed at 2 h, 6 h, 24 h, 3 d and 7 d after reperfusion. Global brain ischemic model was established by 4-VO method. The TdT-mediated dUTP nick end labeling (TUNEL) method was used to detect the neuron apoptosis in hippocampal CA1 region. The immunohistochemical method was applied to investigate the expressions of c-jun and c-fos in hippocampal CA1 region.

RESULTSThe expressions of apoptosis and c-jun and c-fos in CA1 region in S-Sham group, W-I/R group and S-I/R group were more than those in W-Sham group (P < 0.05), were significantly increased in S-I/R group than those in W-I/R group (P < 0.05), and were significantly decreased in S-Cur group than those in S-I/R group (P < 0.05).

CONCLUSIONNeuronal apoptosis and the expressions of c-jun and c-fos are more in SHR hippocampal. Global brain ischemia/reperfusion injury induces more expressions of apoptosis in hippocampal neuron in SHR, and the more expressions of c-jun and c-fos may participate in that process. The neuroprotection of curcumine in SHR is related to c-jun and c-fos.

Animals ; Apoptosis ; drug effects ; Brain Ischemia ; metabolism ; pathology ; CA1 Region, Hippocampal ; drug effects ; metabolism ; Curcumin ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Male ; Neurons ; drug effects ; metabolism ; Proto-Oncogene Proteins c-fos ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Reperfusion Injury ; metabolism ; pathology

OBJECTIVETo create a three dimension (3D) in vitro model for angiogenesis of hemangioma.

METHODThe fragment of hemangioma specimen was embedded in fibrin gel to set up the three-dimension (3D) in vitro model for angiogenesis of hemangioma.

RESULTIn the model, microvessels grew out from the tissue fragments at the 2nd to 3rd day after culture, and at the 8th to 9th day a compact network of microvessels come into being, then tending to be stationary. The compact network around the tissue fragment was confirmed to be blood vessels by immunohistochemistry and electron microscopy.

CONCLUSIONThis model helps to study the mechanism of hemangioma angiogenesis and investigate the drugs of anti-angiogenesis.

Cells, Cultured ; Endothelium, Vascular ; Hemangioma ; Humans ; Models, Cardiovascular ; Neovascularization, Pathologic

BACKGROUNDGenetic factors are the main cause of early miscarriage. This study aimed to investigate aneuploidy in spontaneous abortion by fluorescence in situ hybridization (FISH) using probes for 13, 16, 18, 21, 22, X and Y chromosomes.

METHODSA total of 840 chorionic samples from spontaneous abortion were collected and examined by FISH. We analyzed the incidence and type of abnormal cases and sex ratio in the samples. We also analyzed the relationship between the rate of aneuploidy and parental age, the rate of aneuploidy between recurrent abortion and sporadic abortion, the difference in incidence of aneuploidy between samples from previous artificial abortion and those from no previous induced abortion.

RESULTSA total of 832 samples were finally analyzed. 368 (44.23%) were abnormal, in which 84.24% (310/368) were aneuploidies and 15.76% (58/368) were polyploidies. The first was trisomy 16 (121/310), followed by trisomy 22, and X monosomy. There was no significant difference in the rate of aneuploidy in the advanced maternal age group (≥ 35 years old) and young maternal age group (<35 years old). However, the rate of trisomy 22 and the total rate of trisomies 21, 13, and 18 (the number of trisomy 21 plus trisomy 13 and trisomy 18 together) showed significantly different in two groups. We found no skewed sex ratio. There was no significant difference in the rate of aneuploidy between recurrent miscarriage and sporadic abortion or between the samples from previous artificial abortion and those from no previous artificial abortion.

CONCLUSIONSAneuploidy is a principal factor of miscarriage and total parental age is a risk factor. There is no skewed sex ratio in spontaneous abortion. There is also no difference in the rate of aneuploidy between recurrent abortion and sporadic abortion or between previous artificial abortion and no previous induced abortion.

Abortion, Habitual ; genetics ; Abortion, Spontaneous ; genetics ; Adult ; Aneuploidy ; Female ; Humans ; In Situ Hybridization ; Middle Aged ; Pregnancy ; Sex Ratio

To evaluate the relationship between p27Kip1 low expression in breast cancer and its prognostic implication in breast carcinoma patients. Methods All data that were associated with the study of the relationship between p27Kip1 and the prognosis for breast cancer was pooled from Cochrane library, PubMed, Embase and Medlinebase. The outcome was measured using the risk ratio (RR). Data pooling was performed by RevMan 4. 2. Results 6457 patients from 20 studies were included in this meta-analysis. RR estimate of overall survival (OS) for patients with low level p27Kip1 was 2.07 [1.66,2.60] (P<0.01). For disease free survival (DFS), the pooled RR was 1.27 [1.10,1.47] (P<0.05). The combined RR estimate of relapse free survival (RFS) for patients with low level of p27Kip1 was 1.49 [0.92, 2.42] (P >0.05). In patients with lymph node negative breast carcinoma, the combined RR for OS and RFS were 1.98 [1.34,2.91] (P <0.01) and 1.28 [0.45,3.65] (P > 0.05), respectively. Among the patients with lymph node positive breast carcinoma, the combined RR for OS and RFS was 1.92 [1.31, 2.82] (P=0.0009) and 1.35 [0.96,1.89] (P>0.05) respectively. Conclusions Low level of p27Kip1 appears to be an independent prognostic factor to OS and DFS of breast cancer patients but not to RFS. Additional studies with large patient number and widely accepted practical methods are required to derive the precise prognostic significance of p27Kip1 expression in breast cancer patients.
Biomarkers, Tumor ; analysis ; Breast Neoplasms ; diagnosis ; genetics ; metabolism ; pathology ; Carcinoma ; diagnosis ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; metabolism ; Disease-Free Survival ; Female ; Gene Expression Regulation, Neoplastic ; genetics ; Humans ; Lymphatic Metastasis ; diagnosis ; physiopathology ; Neoplasm Staging ; methods ; Prognosis