Objective To isolate and purify pancreatic ductal epithelial cells in adult rats,and induce differentiation of pancreatic ductal epithelial cells to islets in vitro.Methods By retrograde in- jection of collagenase into biliary-pancreatic tract,pancreatic tissues were digested and different types of pancreatic cells including islets,duct and degranulated aicni cells were separated by means of density gradient centrifugation.Ductal cells were purified by adhering method and identified by immunocyto- chemistry stain of ductal epithelial cells maker antigen(Cytokeratin 19,CK-19).Ductal cells were ex- panded in RMPI 1640 with 10% FBS.About one week when most adherent ceils were of monolayer, the medium was changed to serum-free DMEM/F12 supplemented with keratinocyte growth factor (KGF)to further expand ductal epithelial cells.When ceils reached 80% confluence,nicotinamide and high concentration of glucose were added to promote differentiation of pancreatic ductal epithelial cells.Islets like-structure was stained by Dithizone.Results Irnmunocytochemistry stain of CK-19 re- vealed that most isolated ceils were ductal epithelial ceils.The cultured ductal epithelial cells began to adhere at day 1,reached 80% confluence and cell clones were formed at day 14-21.At day 28,islets- like-structure appeared and was positive for Dithizone staining.Conclusions Ductal epithelial cells of rats can be isolated by means of density gradient centrifugation and purified by adhering method.Duc- tal epithelial cells can differentiate into islets-like-structure in vitro.

The main treatment strategies for chronic pancreatitis in young patients include therapeutic endoscopic retrograde cholangio-pancreatography (ERCP) intervention and surgical intervention. Therapeutic ERCP intervention is performed much more extensively for its minimally invasive nature, but a part of patients are referred to surgery at last. Historical and follow-up data of 21 young patients with chronic pancreatitis undergoing duodenum-preserving total pancreatic head resection were analyzed to evaluate the outcomes of therapeutic ERCP intervention and surgical intervention in this study. The surgical complications of repeated therapeutic ERCP intervention and surgical intervention were 38% and 19% respectively. During the first therapeutic ERCP intervention to surgical intervention, 2 patients developed diabetes, 5 patients developed steatorrhea, and 5 patients developed pancreatic type B pain. During the follow-up of surgical intervention, 1 new case of diabetes occurred, 1 case of steatorrhea recovered, and 4 cases of pancreatic type B pain were completely relieved. In a part of young patients with chronic pancreatitis, surgical intervention was more effective than therapeutic ERCP intervention on delaying the progression of the disease and relieving the symptoms.

Objective To introduce a new technique for urethral coverage in Snodgrass hypospadias repair,and to evaluate its effectiveness and complications.Methods From April 2003 to February 2006, this new procedure was performed in 289 children with hypospadias aged 3 months to 12 years (mean age,2. 4 years).The native meatus of urethra was identified subcoronal in 78 cases,penile/shaft in 136,penoscrotal in 36 and scrotal in 16;and 23 cases had undergoneⅡstage operation and re-operation.The overlapping coverage with bilateral shaft based vascularized dartos pedicle was done in the new urethra by Snodgrass hy- pospadias repair in these children.Results All the cases were followed for 3 months to 2 years.Postoper- atively,urinary fistulas developed in 32 cases (11%).Of them,11 were cured spontaneously within 4 weeks. The incidence of actual urinary fistula was 7% (21/289).Of the 21 fistulas which were not cured,11 (5%) occurred in 214 cases of distal hypospadias;and 10 (13%) in 75 cases of proximal hypospadias,Ⅱstage and re-operation.No dehiscence and diverticulum was found.Combined with mucosal collar technique,the ventral skin of the penis was sewn on the midline.During the follow-up,excellent cosmetic results with normal-ap- pearing circumcised penis were achieved in most patients.Conclusions Bilateral shaft based vascularized dartos pedicle urethral coverage procedure is a reliable and effective method for preventing urethral cutaneous fistulas and dehiscence.This method can reconstruct a satisfactory cosmetic appearance of the penis.

Objective To further evaluate the safety and observe for unknown adverse reactions in the prelicensure trials of Anflu(split influenza virus vaccine)in children aged 6 months to 3 years old and 3 to 11 years old respectively.Methods This open clinical trial enrolled 100 healthy children in each of the two groups:6 months to 3 years old group and 3 to 11 years old group.The 6 months to 3 years group were vaccinated with two pediatric doses(0.25 ml/dose),28 days apart.The 3 to 11 years group were vaccinated with one adult dose(0.5 ml/dose).All the subjects were observed for 30 min after vaccination and had 3 follow-up visits at 24,48,72 h after vaccination. All subjects with adverse reactions were followed up till the symptoms resolved.Results The total adverse reaction rate was 6.0%(12/200).The occurrence rates of local reaction and systemic reaction were 1.0%(2/200)and 5.5%(10/200)respectively.For the younger group and older group,the adverse reaction rates were 8.0%and 4.0%respectively.Conclusion This vaccineis safe in children aged 6 months to 11 years old.

Objective To further evaluate the safety and observe for unknown adverse reactions in the prelicensure trials of Anflu(split influenza virus vaccine)in children aged 6 months to 3 years old and 3 to 11 years old respectively.Methods This open clinical trial enrolled 100 healthy children in each of the two groups:6 months to 3 years old group and 3 to 11 years old group.The 6 months to 3 years group were vaccinated with two pediatric doses(0.25 ml/dose),28 days apart.The 3 to 11 years group were vaccinated with one adult dose(0.5 ml/dose).All the subjects were observed for 30 min after vaccination and had 3 follow-up visits at 24,48,72 h after vaccination. All subjects with adverse reactions were followed up till the symptoms resolved.Results The total adverse reaction rate was 6.0%(12/200).The occurrence rates of local reaction and systemic reaction were 1.0%(2/200)and 5.5%(10/200)respectively.For the younger group and older group,the adverse reaction rates were 8.0%and 4.0%respectively.Conclusion This vaccineis safe in children aged 6 months to 11 years old.

BACKGROUNDTetralogy of Fallot (TOF) is the most common malformation of children with an incidence of approximately 10% of congenital heart disease patients. There can be a wide spectrum to the severity of the anatomic defects, which include ventricular septal defect, aortic override, right ventricular outflow tract obstruction, and right ventricular hypertrophy. We examined the relationship between right ventricular hypertrophy in patients with TOF and the gene expression of factors in the mitogen-activated protein kinase (MAPK) signal pathway.

METHODSTo gain insight into the characteristic gene(s) involved in molecular mechanisms of right ventricular hypertrophy in TOF, differential mRNA and micro RNA expression profiles were assessed using expression-based micro array technology on right ventricular biopsies from young TOF patients who underwent primary correction and on normal heart tissue. We then analyzed the gene expression of the MAPK signal pathway using reverse transcription-polymerase chain reaction (RT-PCR) in normals and TOF patients.

RESULTSUsing the micro RNA chip V3.0 and human whole genome oligonucleotide microarray V1.0 to detect the gene expression, we found 1068 genes showing altered expression of at least two-fold in TOF patients compared to the normal hearts, and 47 micro RNAs that showed a significant difference of at least two-fold in TOF patients. We then analyzed these mRNAs and micro RNAs by target gene predicting software Microcosm Targets version 5.0, and determined those mRNA highly relevant to the right ventricular hypertrophy by RT-PCR method. There were obvious differences in the gene expression of factors in the MAPK signal pathway when using RT-PCR, which was consistent to the results of the cDNA microarray.

CONCLUSIONThe upregulation of genes in the MAPK signal pathway may be the key events that contribute to right ventricular hypertrophy and stunted angiogenesis in patients with TOF.

Child, Preschool ; Humans ; Hypertrophy, Right Ventricular ; genetics ; In Vitro Techniques ; Male ; MicroRNAs ; Mitogen-Activated Protein Kinases ; genetics ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; genetics ; physiology ; Tetralogy of Fallot ; genetics

OBJECTIVETo investigate the method of auricular reconstruction for concha-type microtia.

METHODSTwo-staged auricular reconstruction was applied in 13 cases (14 ears) with concha-type microtia. The cartilage auricular framework was fabricated and implanted in the first stage, followed by ear elevation and cranio-auricle angle formation at the second stage.

RESULTSThe patients were followed up for 2 months to 2 years with satisfactory aesthetic result. The reconstructed ears had a good appearance and position, and were symmetric to the healthy ears.

CONCLUSIONSThe two-staged auricular reconstruction with autologous cartilage framework is ideal for concha-type microtia.

Adolescent ; Adult ; Cartilage ; transplantation ; Child ; Ear Auricle ; surgery ; Ear, External ; abnormalities ; Female ; Humans ; Male ; Reconstructive Surgical Procedures ; Ribs ; Tissue Scaffolds ; Transplantation, Autologous ; Young Adult

The main treatment strategies for chronic pancreatitis in young patients include therapeutic endoscopic retrograde cholangio-pancreatography (ERCP) intervention and surgical intervention. Therapeutic ERCP intervention is performed much more extensively for its minimally invasive nature, but a part of patients are referred to surgery at last. Historical and follow-up data of 21 young patients with chronic pancreatitis undergoing duodenum-preserving total pancreatic head resection were analyzed to evaluate the outcomes of therapeutic ERCP intervention and surgical intervention in this study. The surgical complications of repeated therapeutic ERCP intervention and surgical intervention were 38% and 19% respectively. During the first therapeutic ERCP intervention to surgical intervention, 2 patients developed diabetes, 5 patients developed steatorrhea, and 5 patients developed pancreatic type B pain. During the follow-up of surgical intervention, 1 new case of diabetes occurred, 1 case of steatorrhea recovered, and 4 cases of pancreatic type B pain were completely relieved. In a part of young patients with chronic pancreatitis, surgical intervention was more effective than therapeutic ERCP intervention on delaying the progression of the disease and relieving the symptoms.
Adolescent ; Adult ; Cholangiopancreatography, Endoscopic Retrograde ; adverse effects ; methods ; Diabetes Mellitus ; etiology ; Female ; Follow-Up Studies ; Humans ; Male ; Pain, Postoperative ; etiology ; Pancreatectomy ; adverse effects ; methods ; Pancreatitis, Chronic ; surgery ; Postoperative Complications ; etiology ; Steatorrhea ; etiology ; Treatment Outcome ; Young Adult

OBJECTIVETo evaluate some currently used surgical approaches to the correction of congenital penile curvature.

METHODSSeventy-six patients with congenital penile curvature underwent surgical correction, of whom 67 were accompanied with hypospadias, 5 with epispadias and 4 with normal urethral meatus. The methods for straightening the phallus included 5-week preoperative hCG treatment, complete degloving of penile skin, release of periurethral fibrous bands extending proximally to the meatus, plication of dorsal or ventral tunica albuginea, and embedding of dermis and tunica vaginalis grafts.

RESULTSAll the cases were followed up for 2 months to 2 years, and the mean follow-up time was 9.3 months. Satisfactory phallus straightening was achieved in 67 cases (88%), mild residual chordee remained in 6 (8%), which needed no reoperation for the time being unless warranted by follow-up, and relapse occurred in 3 (4%), which needed further operation.

CONCLUSIONMost cases of congenital penile curvature can be corrected sufficiently with the above methods.

Child ; Child, Preschool ; Follow-Up Studies ; Humans ; Infant ; Male ; Penis ; abnormalities ; Urogenital Abnormalities ; surgery ; Urologic Surgical Procedures, Male ; methods

OBJECTIVETo investigate the expression of integrin beta 1 in hepatic cirrhosis (HC) and hepatocellular carcinoma (HCC).

METHODSThe expression of integrin beta 1 in HCC, HC and normal liver tissues was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and laser scanning confocal microscopy (LSCM). The association between the integrin beta 1 expression and clinical pathological features were analyzed.

RESULTS(1) The levels of integrin beta 1 mRNA and protein in the HCC (1.30+/-0.24, 90.50+/-33.50) and HC (1.58+/-0.31, 123.10+/-38.90) were much higher than that in the normal hepatic tissue (0.37+/-0.08, 11.90+/-6.00) (P less than 0.05). (2) The expression of integrin beta 1 was associated with HC (r = 0.692), Edmondson pathologic grade (F = 13.618), encapsulation (F = 17.857) and metastasis (F = 38.857) (P less than 0.01).

CONCLUSIONSIntegrin beta 1 may play an important role in the development of hepatic fibrosis, hepatic cirrhosis and hepatocellular carcinoma.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Humans ; Integrin beta1 ; genetics ; metabolism ; Liver ; metabolism ; Liver Cirrhosis ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics