Objective To investigate the expression of human bone morphogenetic protein-7(hBMP-7)in rat bone malTOW stromal cells(BMSC)with a recombinant adenovirai vector carrying the hBMP-7 gene(Ad-hBMP-7) and study the effecta of Ad-hBMP-7 transfection on BMSC difierentiation.in order to explore the possibility for hBMP-7 gene therpy.Methods The rat BMSC cultured in vitro.They were divided into 3 groups:untreated group,Ad-hBMP-7 and Ad-GFP transduced treated group.The rat BMSC were transfected by Ad-hBMP-7 and Ad-GFP. The expression of hBMP-7 was detected by RT-PCR and Western blot analysis.and the alkaline phosphatage (ALP)activity of the BMSC was observed.ResulIs In the Ad-hBMP-7 transduced treated group.hBMP-7 mRNA expression wag manifested detected by RT-PCR(470 bp),Westem blot analysis demonstrated that these cells indeed produced the hBMP-7 protein(Mr.15×103);10 days after transduction treatment,most of the BMSC were had brown black particles stained positively by ALP activity.But in Ad-GFP transduced treated group and untreated group they did not.Conclusions Ad-hBMP-7 could efficiently transfect BMSC and promote the conversion to osteoblast.The expression of hBMP-7 in rat BMSC provides a basis for hBMP-7 gene therapv.

Objective To analysis and determine the possibility of the Citellus undulatus infected with Yersinia pestis surviving the winter in an experimental study, and to provide scientific experimental basis for the study on the mechanism of Yersinia pestis preservation. Method In 2006,09 to 2007,04 and 2007,09 to 2008,04 in Xinjiang Wusu-Gurtu natural foci of plague, under natural conditions, the over the winter process of Citellus undulatus carrying the plague bacteria was simulated, and 178 Citellus undulatus were infected with Yersinia pestis (1×107 Bacteria/mouse) using artificial injection method. One hundred seventy-eight Citellus undulatus infected with Yersinia pestis were kept into a construction of the black (1-5 ℃) basement (2 meters under the ground) in the plague focus. In doing so, these Citellus undulatuses almost simultaneously stepped into hibernation. After waking up from hibernation in following year in April, the survived mice carrying the plague bacteria were observed. Results Sixty-eight mice survived among the 178 infected with Yersinia pestis after 6 months of hibernation (through October to the following year in April), and the remaining 110 were all dead without pulling through the hibernation period. The survival rate was 38.2% (68/178). The organ culture of Yersinia pestis of the 110 dead mice(Citellus undnlatus) were tested, 67 were negative(-), 43 positive(+), with a positive rate of 39.1%(43/110). Among the rats with positive plague bacteria, the congestive pulmonary edema and the pathological changes of the hemorrhagic inflammation of the heart, liver, spleen, kidney and injection site could be seen clearly; the plague-free mice were not found to have any pathological changes. The survived 68 mice over the winter were autopsied and observed after being fed up for 20 days. No any pathological changes were found among these mice, and culturing of Yersinia pestis of the heart, liver, spleen, lungs and the tissue of injection site of these mice were all negative (-). Conclusions Citellus undulatus can carry Yersinia pestis during hibernation, but some fail to carry the bacteria through the entire process of hibernation persistently. Yersinia pestis was negative in the survived mice at the end of hibernation. The results showed that Citellus undulatus can not carry Yersinia pestis over the winter.

OBJECTIVETo investigate the effectiveness of surgical strategies for Shang Ring circumcision in the treatment of short frenulum praeputii in patients with redundant prepuce or phimosis.

METHODSTotally, 130 cases of short frenulum praeputii with redundant prepuce or phimosis were randomly assigned to an experimental group and a control group of equal number to receive Shang Ring circumcision, the former by transverse incision in the distal penis foreskin and pull-up of the interior board, and the latter by conventional transverse incision and longitudinal suture of the frenulum praeputii. Comparisons were made between the two groups in the surgical duration, intraoperative blood loss, 24 h postoperative pain visual analog score (VAS), postoperative complications, satisfaction with the penile appearance, and the quality of sexual life.

RESULTSThe surgical duration, intraoperative blood loss, 24 h postoperative VAS, postoperative sexual satisfaction, and satisfaction with penile appearance were (4.60 +/- 1.20) min, (2.61 +/- 1.81) ml, 1.73 +/- 0.76, 98.5%, and 98.5%, respectively, in the experimental group, as compared with (21.60 +/- 6.30) min, (11.10 +/- 3.40) ml, 5.37 +/- 1.84, 70.3% and 69.8% in the control, with statistically significant differences between the two groups (P < 0.05). The incidence rates of such major complications as wound dehiscence, infection, and moderate to severe edema were 1.5% (1/65), 3.1% (2/65), and 4.6% (3/65), respectively, in the experimental group in comparison with 12.3% (8/65), 15.3% (10/65), and 30.7% (20/65) in the control, with statistically significant differences between the two groups (P < 0.05). None of patients had any serious complications.

CONCLUSIONShang Ring circumcision by transverse incision in the distal penis foreskin and pull-up of the interior board, with its advantages of shorter operation time, less blood loss, mild pain, fewer complications, and higher satisfaction and acceptance of the patients, can be used as an safe and effective approach to the treatment of short frenulum praeputii.

Aged ; Blood Loss, Surgical ; statistics & numerical data ; Circumcision, Male ; adverse effects ; instrumentation ; methods ; Edema ; epidemiology ; Foreskin ; abnormalities ; surgery ; Humans ; Incidence ; Male ; Operative Time ; Pain Measurement ; Pain, Postoperative ; diagnosis ; Patient Satisfaction ; Phimosis ; surgery ; Postoperative Period ; Prostheses and Implants ; Surgical Wound Dehiscence ; epidemiology ; Surgical Wound Infection ; epidemiology

Objective To improve the awareness of fetal cardiac rhabdomyomas (CRs) and investigate a better model for prenatal diagnosis and treatment through analyzing imaging findings and prognosis.Methods A retrospective study was conducted on 23 cases of CRs which were diagnosed by ultrasound in Obstetrics and Gynecology Hospital of Fudan University from January 2008 to November 2015.General conditions,imaging features,prognosis and follow-up data of the 23 cases were described.Results The average gestational age of the 23 fetuses at diagnosis was (29.8±4.1) (22.4-35.7) weeks.Seventeen out of the 23 gravidas received prenatal multidisciplinary consultation.Among all 23 gravidas,three (13％) were lost to follow-up,12 (52％) decided to terminate the pregnancy,and the other eight (35％) continued to term pregnancy and their babies were followed up for three years.Of these eight cases,two cases received prenatal brain MRI and no tuberous sclerosis complex (TSC) was detected,no CRs was identified during the follow-up,and their physical and mental developments were both normal.One case was diagnosed with suspected subependymal nodules by prenatal brain MRI in our hospital,but the MRI images was normal when scanned in the other hospital,and follow-up data revealed neither CRs nor abnormal physical and mental developments.Four cases did not received prenatal brain MRI,but the MRI images of neonatal brains indicated TSC,besides,follow-up data showed that seizures were observed,physical developments were all normal,but three of the four cases had mental retardation;CRs disappeared in only two of the four cases.One case had neither prenatal nor neonatal MRI,but follow-up data showed that CRs had disappeared and physical and mental developments were both normal.Conclusions Prenatal diagnosis of fetal tuberous sclerosis is crucial to the prognosis of CRs.Prenatal ultrasonography in combination with cranial MRI improves the accuracy of prenatal diagnosis of CRs complicated with TSC and assists in clinical decision-making and prognosis analysis.

OBJECTIVETo investigate the expression and activity of fatty acid amide hydrolase (FAAH) in the colon and its role in children with slow transit constipation (STC).

METHODSPatients were divided into constipation group (n=21) and control group (n=15). The constipation group was consistent with the diagnostic criteria for STC. Western blotting, immunohistochemistry and real-time PCR were used to examine the FAAH expression in surgical specimen of colon. The location and distribution of FAAH and cannabinoid receptor type 1 (CB1) were detected by immunofluorescence double staining. The biological activity of colon FAAH was detected by high-performance liquid chromatography.

RESULTSWestern blotting revealed that FAAH protein expression in the ascending colon, descending colon and sigmoid colon were significantly decreased in the myenteric neurons and absorption cells in the constipation group as compared with the control group (8.68+/-3.4 vs 10.47+/-3.7, 8.21+/-1.2 vs 9.95+/-6.4, 8.01+/-7.2 vs 9.79+/-3.4, all P<0.05). The same results were found by immunohistochemistry and real-time PCR. The FAAH hydrolysis activity in the ascending colon, descending colon or sigmoid colon decreased significantly in the constipation group, as compared with the control group [(0.51+/-0.23) nmol x min(-1) x mg(-1) vs (0.84+/-0.24) nmol x min(-1) x mg(-1), (0.39+/-0.25) nmol x min(-1) x mg(-1) vs (0.55+/-0.44) nmol x min(-1) x mg(-1), (0.35+/-0.37) nmol x min(-1) x mg(-1) vs (0.58+/-0.48) nmol x min(-1) x mg(-1), all P<0.05].

CONCLUSIONFAAH expression and hydrolysis activity in the colon decrease in children with STC. FAAH may play a role in the pathogenesis of slow transit constipation in Children.

Adolescent ; Amidohydrolases ; metabolism ; Case-Control Studies ; Child ; Child, Preschool ; Chronic Disease ; Colon ; metabolism ; Constipation ; metabolism ; Female ; Humans ; Male

OBJECTIVETo construct a recombinant adenoviral vector carrying HBcAg-HSP70 chimeric gene by homologous recombination in bacteria and to detect its expression in vitro.

METHODSHeat shock protein 70 gene from Mycobacterium tuberculosis were amplified by PCR and were cloned to adenoviral shuttle plasmid pAdTrack-CMV-HBsAg. Then the resultant pAdTrack-CMV-HBsAg-HSP70 was cotransfected into BJ5183 bacteria with the plasmid pAdeasy-1. The adenoviral plasmid carrying HBsAg-HSP70 gene (pAd-HBsAg-HSP70) was generated with homologous recombination in bacteria and the adenoviruses were produced in 293 cells. Several kinds of mammal cells (293 cells and Vero cells) were infected with adenoviruses and the expression of HBsAg-HSP70 was detected by RT-PCR and ELISA in vitro.

RESULTSThe adenoviral plasmids pAd-HBsAg-HSP70 were obtained by selection for kanamycin resistance and confirmed by restriction endonuclease Pac analyses. The recombinant adenoviruses Ad-HBsAg-HSP70 were packaged successfully in 293 cells. The titer of Ad-HBsAg-HSP70 was up to 2 x 10(12) pfu/L after the second passage of proliferation in 293 cells. HBsAg and HSP70 were expressed efficiently in mammal cells after infection.

CONCLUSIONThe recombinant adenoviruses expressing HBsAg and HSP70 were constructed successfully which can be used further in study of gene therapy for HBV.

Adenoviridae ; genetics ; Animals ; Cell Line ; Cercopithecus aethiops ; Defective Viruses ; genetics ; Enzyme-Linked Immunosorbent Assay ; Green Fluorescent Proteins ; genetics ; metabolism ; HSP70 Heat-Shock Proteins ; genetics ; metabolism ; Hepatitis B Surface Antigens ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Vero Cells ; Virus Replication ; genetics

OBJECTIVETo explore whether hepatitis B virus (HBV) S gene-modified dendritic cells (DCs) might induce a specific cytotoxic T lymphocyte (CTL) response.

METHODSThe recombinant adenoviruses carrying HBsAg genes were prepared and used to transfect DCs generated from cord blood. The efficacy of transfection was observed through the expression of enhanced green fluorescent protein (EGFP) in DCs and the expression of HBsAg was detected by ELISA. HBV S gene-modified DCs were co-cultured with T cells from cord blood and T cells stimulating activities were detected using mixed lymphocyte reaction (MLR). The CTL assay was carried out to assess the ability of CTL lines to lyse target cells of HepG(2)22.1.5 by measuring lactate dehydrogenase (LDH) release.

RESULTSThe results showed that HBV S genes were expressed in DCs with high efficacy by recombinant adenoviral vector. DCs had a normal shape after transfection. The result of MLR showed that HBV S gene-modified DCs could effectively stimulate naive T cells to proliferate. The induced specific CTL lines could lyse target cells of HepG(2)22.1.5.

CONCLUSIONSHBV S gene-modified DCs enhanced the function to induce a specific CTL effect, showing its promise for developing anti-viral vaccine in future.

Cell Line ; Cells, Cultured ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; virology ; Hepatitis B ; immunology ; virology ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B virus ; genetics ; immunology ; Humans ; Lymphocyte Culture Test, Mixed ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo study HBV preS2/S gene expression effects in mammalian cells transferred with recombinant adenoviral vector.

METHODSThe replication-deficient recombinant adenoviral vector (Ad-HBs) carrying HBV preS2/S gene were constructed by homologous recombination in bacteria. The 293 cells, Vero cells, HepG2 cells and mesenchymal stem cells (MSCs) were infected with adenoviruses. The expressions of enhanced green fluorescent protein (EGFP) were observed with fluorescence microscope and the expressions of HBsAg were detected by RT-PCR and ELISA in vitro.

RESULTSMore than 90% of 293 cells, Vero cells, HepG2 cells or MSCs expressed EGFP after transfection at the MOI of 20 and the titers of HBsAg were more than 3.229 (A value) in culture supernatant.

CONCLUSIONThe HBV preS2/S gene was not only expressed efficiently in immortalized cells, but also expressed efficiently in stem cells with the recombinant adenoviruses vector.

Adenoviridae ; genetics ; Animals ; Cell Line ; Cell Line, Tumor ; Cercopithecus aethiops ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Hepatitis B Surface Antigens ; genetics ; metabolism ; Hepatitis B virus ; genetics ; immunology ; Humans ; Microscopy, Fluorescence ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Vero Cells

OBJECTIVETo observe the clinical effects of the no-flip procedure with the Chinese Shang Ring when circumcising adult males with redundant prepuce or phimosis, and to discuss its advantages and disadvantages.

METHODSUsing the no-flip Shang Ring technique, we performed circumcision for 167 adult males aged 18 -72 (mean 27.8) years with redundant prepuce or phimosis, and analyzed the clinical data, including the operation time, postoperative complications, ring-removal time, and postoperative appearance of the penis.

RESULTSComplete follow-up data of 94 cases (56.29%) were obtained. The mean operation time was (5.03 +/- 0.71) minutes and the average ring-removal time was (18.83 +/- 6.70) days. The primary postoperative complications were edema (35 cases [37.23%] at 2 weeks and 9 cases [9.57%] at 4 weeks), including 2 severe cases (2.13%), and infection (3 cases [3.19%]). The pain scores were 2.01 +/- 2.46 during the procedure and 4.52 +/- 2.53 at 24 hours postoperatively. Slipping of the outer ring occurred in 1 case, and delayed removal of the ring in 30 cases (31.91%).

CONCLUSIONAdult male circumcision with the no-flip Shang Ring technique is recommended for its short operation time, simple procedure, fewer postoperative complications, less pain, and better incision appearance.

Adult ; Aged ; Circumcision, Male ; adverse effects ; instrumentation ; methods ; Edema ; etiology ; Humans ; Male ; Middle Aged ; Operative Time ; Pain, Postoperative ; etiology ; Penile Diseases ; etiology ; Penis ; abnormalities ; surgery ; Phimosis ; surgery ; Postoperative Complications ; Postoperative Period ; Prostheses and Implants ; Young Adult

OBJECTIVETo analyze the spatial distribution of highly pathogenic avian influenza (HPAI) and to explore environmental factors associated with HPAI using geographic information system (GIS) techniques in Mainland China.

METHODSDatabases were set up using the information of HPAI during epidemics in 2004, and linked to digital maps at provincial and county administrative layers in the country through the ArcGIS 8.3 software. Spatial cluster analyses, spatial statistics analyses and tracking analyses on epidemic situation of HPAI were implemented. Environmental factors associated with HPAI were also analyzed on data related to weather, vegetation and migratory birds etc.

RESULTSFindings from spatial cluster analyses showed that high incidence area was centralized in 113.261 degrees ordm; east longitude and 23. 119 degrees ordm; north latitude with a radius of 1090.52 kilometers (relative risk= 2.646, P value= 0.001). Spatial statistical analyses showed that HPAI took place mainly in capital cities of provinces and surrounding areas as well as in the circumference areas of arterial rivers, lakes and seacoasts. Results also showed that HPAI occurrences were associated with low air temperature, high relative humidity and high air pressure as well as with east & central migration routes of migratory birds. The average normalized difference vegetation index was 0.36 +/- 0.11 in epidemic areas of HPAI.

CONCLUSIONHPAI was unrandomly distributed and geographically clustered in China.

Animal Migration ; Animals ; Atmospheric Pressure ; Birds ; virology ; China ; epidemiology ; Cluster Analysis ; Environment ; Geographic Information Systems ; Humidity ; Influenza A Virus, H5N1 Subtype ; pathogenicity ; Influenza in Birds ; epidemiology ; Temperature