Objective To investigate the dose-effect relationship of donor antigenic specificity CD4~+ CD25~+ Treg cells prolonging the survival of rat kidney allograft.Methods Sixty allograft kid- ney transplantation animal models were established with SD rats as donors and Wistar rats as recipi- ents.CD4~+ CD25~+ T cells were sorted from Wistar rats'spleens by way of MACS and the phenotypes of donor antigenic specificity were induced in vitro.According to the quantities of prepared CD4~+ CD25~+ T cells injected through tail vein in kidney transplantation,models were divided into four ex- perimental groups:2?10~5(group I),5?10~5(groupⅡ),1?10~6(groupⅢ),2?10~6(groupⅣ).The models which had no injection served as control group(n=12).The survival of the transplanted kid- ney was observed.The levels of blood serum creatinine were determined and the histopathological changes in the transplanted kidney were observed at day 4,9 and 15.The results of histopathology were evaluated according to the standard of Banff Schema and the semi-quantitative scores were gained by way of Watanabe.The reaction indexes of receptor spleen cells to the donor antigens were tested by way of MTT at day 15.Results The mean survival time of transplanted kidneys was the highest in groupⅢ[(31.4?4.6)days]and lowest in the control group[(11.7?6.2)days].There was signif- icant difference between groupⅢand control group in levels of serum cre'atinine(P

Objective To evaluate the in vitro antiviral activity of HB-13,a compound derivant from berberine and its prodrug berberine,against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2). Methods Vero cells were cultured in vitro and infected with HSV.Then,various concentrations of HB-13, berberine,and aciclovir were used to treat these infected cells.The cytopathic effect was observed to deter- mine the antiviral effects and cytotoxicity of HB-13 and berberine.Results For HB-13,berberine and acy- clovir,the half toxicity concentration (TC_(50)) to Vero cells was 31.99,380 and more than 800?g/mL, respectively;the average half inhibitory concentration (IC_(50)) against HSV-1 was 1.328,more than 100,and 0.443?g/mL,respectively,the treatment index (TI) against HSV-1 was 24.09,less than 3.80,and more than 1805.87,respectively;the IC_(50) against HSV-2 was 1.344,more than 100,and 0.679?g/mL,respectively,the TI against HSV-2 was 23.80,less than 3.80 and more than 1178.20,respectively.Conclusion HB-13 possesses marked antiviral activity against HSV-1 and HSV-2 in vitro,while berberine does not.

Objective To explore the clinical feature,diagnosis and treatment of postoperative chylothorax after congenital cardiac surgery.Methods From Jan.2000 to Jan.2008,36 consecutive infants following cardiac surgery were diagnosed postoperative chylothorax.Age at operation ranged 6 days to 4 years.Weight ranged 3.5 kg to 17.0 kg.Indications for cardiac surgery were:tetralogy of Fallot 18 cases,complete atrioventricular septal defect 5 cases,double outlet of right ventricle 4 cases,complex heart defect for Fontan operation 4 cases,coarctation of the aorta 2 cases,total anomalous pulmonary venous drainage 2 cases,patent ductus arteriosus 1 case.Chylothorax occurred in right side was 32 cases,in left side 3 cases,bilateral side 1 case.Diagnostic time for chylothorax was postoperative 1-10 d.Results Thirty-four cases were treated conservatively with success(34/36,94.44%).Only 2 cases needed surgical treatment.The chest drainage volume before con-servative treatment was 22-60 mL/(kg?d)[mean 40.82 mL/(kg?d)].The mean duration of chylous drainage was 15 d(ranged 7 d to 42 d).There was no death in hospital.Two children had sepsis.The cases were followed up 2 weeks-7 years.One newborn was abandoned because of the other reason after discharge.There was no recurrence of chylothorax.Twenty-two cases were still in low-fat diet.Conclusions The chylothorax following operation of congenital heart disease should be noticed and early diagnosed.The initial treatment is conservative,but surgical treatment ought to be considered if the conservative management is unsuccessful.J Appl Clin Pediatr,2009,24(1):70-72

Objective To study the clinical feature,treatment,and prognosis of the cytomegalovirus (CMV)pneumonia patients treated with immunosuppressor against kidney disease.Mlethod The patients received immunosuppressor against kidney disease in The First Affiliated Hospital of Sun Yat-sen University from June 1999 to December 2006.CMV antigen of leucocyte in the peripheral blood and/or bronchoalveolar lavage fluid of these patients were detected with immunocytochemical methods,and 21 patients were found suffering from CMV pneumonia.The 21 patients were introvenously injected with ganciclovir 5～10 mg/(kg?d),and the immunosuppressive agent treatment suspended.Their clinical feature and prognosis were retrospectively analyzed. Results The 21 patients received corticosteroids before CMV pneumonia contracted,of them,13 patients had been intensively treated with Methyllprednisolone with mean total dose(3.2?0.6)g.Of them,15 had been treated with cyclophosphamide with mean total dose(3.8?1.3)g.The median time from the beginning of using immunosuppressor to the onset of CMV pneumonia was 25(13～92)days.All patients had fever,cough, shortness of breath and X-ray showed interstitial pneumonia,of them,19 patients developed hypoxemia,and 11 patients' CMV antigen was positive in the leucocyte from bronchial lavage fluid.The result showed 9 patients survived and 12 died.The average duration of treatment with ganciclovir was(26.2?6.3)days. CMV pneumonia is a serious complication in patients who were treated with immunosuppressor against kidney disease.The mortality is high.Ganciclovir is a medicine of choice to treat CMV pneumonia.

OBJECTIVETo present the methods of vacuum sealing drainage and free coupling chain-link flap of posterior tibial artery flap and medial plantar flap in the reconstruction of degloving injury of propodium.

METHODSFrom Oct. 2008 to Dec. 2011 five cases with degloving injury of propodium underwent debridement and vacuum sealing drainage on the first stage. Free chain-link flap of posterior tibial artery flap and medial plantar flap were applied to close the wound at the secondary stage. The nerve was included in the coupling flaps. The size of posterior tibial artery flap ranged from 14 cm x 10 cm to 11 cm x 8 cm,and the size of medial plantar flap ranged from 12 cm x 8 cm to 8 cm x 6 cm.

RESULTSAll flaps were survived with no vascular crisis. The flap sensation recovered to S3-S3 during the follow-up period of 6-21 months. The texture and appearance of flaps were satisfied. The plantar had not ulcer and corpus callosum.

CONCLUSIONVacuum sealing drainage and free chain-link flap of posterior tibial artery flap and medial plantar flap with nerve are the ideal methods for the reconstruction of degloving injury of propodium.

Adult ; Drainage ; methods ; Female ; Foot Injuries ; surgery ; Free Tissue Flaps ; blood supply ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Tibial Arteries ; transplantation ; Vacuum ; Young Adult

Betulinic acid is a naturally occurring pentacyclic triterpenoid, which has antiretroviral, antimalarial, and anti-inflammatory properties. The purpose of this study is to investigate the HBV DNA replication inhibition in the mouse model with betulinic acid. Hydrodynamic injection method via the tail vein with the Paywl. 3 plasmid was used to establish the animal mode (n = 15), and the mice were randomly divided into the PBS control group (n = 5), Betulinic acid treatment group (n = 5) and lamivudine control group (n = 5). The day after successful modeling , the mice would have taken Betulinic acid (100 mg x kg(-1)), lamivudine (50 mg x kg(-1)), PBS drugs orally, once daily for 7 days, blood samples were acquired from the orbital venous blood at 3, 5, 7 days after the administering, HBsAg and HBeAg in serum concentration were measured by ELISA and the mice were sacrificed after 7 days, HBV DNA southern detections were used with part of mice livers. The results showed that betulinic acid significantly inhibited the expression of HbsAg in the mice model at the fifth day compared with the control group, and there was no significant differences between the effects of lamivudine and the PBS control group; both the betulinic acid and lamivudine groups had no significant inhibition for the HBeAg expression; the HBV DNA expressions of the liver tissue from the betulinic acid and lamivudine groups were inhibited compared with the control group. Taken together, these results reveal betulinic acid can inhibit the HBsAg expression and replication of the liver HBV DNA in the mouse model.
Acute Disease ; Animals ; Antiviral Agents ; pharmacology ; DNA Replication ; drug effects ; DNA, Viral ; biosynthesis ; Hepatitis B ; blood ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; drug effects ; genetics ; immunology ; physiology ; Male ; Mice ; Plasmids ; genetics ; Triterpenes ; pharmacology ; Virus Replication ; drug effects

OBJECTIVETo examine the expression of vascular endothelial growth factor (VEGF) and its receptors (VEGFR1, VEGFR2) in transdifferentiated human proximal tubular epithelial (HK-2) cell induced by transforming growth factor beta1 (TGFbeta1).

METHODSThe transdifferentiation of HK-2 cells was detected by evaluation of expression of alpha-SMA by cytoimmunochemistry and RT-PCR. The VEGF mRNA was evaluated with RT-PCR. The secreted VEGF in the culture media was measured with ELISA. The cellular VEGF, VEGFR1, and VEGFR2 were measured with Western blot.

RESULTSThe immunostain of alpha-SMA were positive in HK-2 cell induced by TGFbeta1 at the concentration of 5 and 8 ng/ml for 72 h. The expression of alpha-SMA mRNA was induced by TGFbeta1 in concentration- and time-dependent manners. The expressions of mRNA and protein of VEGF were upregulated by TGFbeta1 at the concentration of 0.1 and 1 ng/ml for 72 h and at the concentration of 8 ng/ml for 12 h and 24 h when compared with the control. But expressions of mRNA and protein of VEGF were downregulated by TGFbeta1 at the concentration of 3, 5, and 8 ng/ml for 72 h and at the concentration of 8 ng/ml for 36, 48, and 72 h, respectively. Meanwhile, Protein levels of VEGFR1 and VEGFR2 were upregulated by TGFbeta1 in concentration- and time- dependent manners.

CONCLUSIONSIncreased expression of VEGFR1 and VEGFR2 and two-phase change in VEGF expression occurred in the process of tubular epithelial transdifferentiation induced by TGFbeta1. Reduced expression of VEGF may contribute to tubular epithelial transdifferentiation in a vicious circle.

Cell Differentiation ; Epithelial Cells ; cytology ; Humans ; Kidney Tubules, Proximal ; cytology ; RNA, Messenger ; metabolism ; Receptors, Vascular Endothelial Growth Factor ; metabolism ; Transforming Growth Factor beta ; pharmacology ; Transforming Growth Factor beta1 ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

OBJECTIVETo analyze genomic copy number variations in an infant with Cri du Chat syndrome, and to explore the underlying genetic cause.

METHODSG-banding analysis was carried out on cultured peripheral blood sample from the patient. Copy number variation analysis was performed using microarray comparative genomic hybridization, and the result was verified with fluorescence in situ hybridization.

RESULTSThe infant was found to have a 46, XY, der(5) (p?) karyotype. By microarray comparative genomic hybridization, a 23.263 Mb deletion was detected in 5p14.2-p15.3 region in addition to a 14.602 Mb duplication in 12p31 region. A derivative chromosome was formed by rejoining of 12p31 region with the 5p14.2 breakpoint. The patient therefore has a karyotype of arr cgh 5p15.3p14.2 (PLEKHG4B>CDH12)× 1 pat, 12p13.33p13.1 (IQSEC3>GUC Y2C)× 3 pat. Loss of distal 5p and gain of distal 12p were verified with fluorescence in situ hybridization.

CONCLUSIONThe Cri du Chat syndrome manifested by the patient was caused by deletion of distal 5p from an unbalanced translocation involving chromosome 5. Microarray comparative genomic hybridization is a powerful tool for revealing genomic copy number variations for its high-resolution, high-throughput and high accuracy.

Adult ; Chromosome Banding ; Chromosome Deletion ; Comparative Genomic Hybridization ; Cri-du-Chat Syndrome ; genetics ; DNA Copy Number Variations ; Female ; Humans ; Infant ; Male

OBJECTIVETo report the outcome of long bone nonunion of humerus, femur and tibia treated with locking internal fixation and bone graft.

METHODSFrom February 2003 to October 2006, locking internal fixation and bone grafting were employed to treat 5 cases at humerus, 33 cases at femur, 23 cases at tibia. Forty-four of the patients were men, and 17 were women. The mean age was 38 years (range 7-70 years). The nonunion had resulted from failure of internal fixation in 47 cases, failure of external fixation in 5 cases, infection in 9 cases. The history of nonunion lasted from 10 to 156 months (mean 19 months). There were 42 patients treated with locking compression plate (LCP), and 19 patients with less invasive stabilization system (LISS). For bone grafting, autogenous ilium was used in 55 patients, autogenous ilium and allograft bone was used in 3 patients, allograft bone and Wright DBM artificial bone was used in 3 patients.

RESULTSAll the 61 patients were followed up for an average 12 months (range 6-24 months) only to reveal solid bone union in all the fracture, with a mean healing time of 4.8 months (ranged from 4 to 6 months). No loosening or breakage of the implants occurred in this series. The Knee Society Scores (KSS) was used to evaluate knee function in 47 patients with peri-knee joint nonunion, excellent result were seen in 35 patients, good in 7 patients, fare in 1 patients, poor in 4 patients.

CONCLUSIONLocking internal fixation can be used to treat effectively bone nonunion at the humerus, femur and tibia.

Adolescent ; Adult ; Aged ; Bone Plates ; Child ; Female ; Femoral Fractures ; surgery ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Fractures, Ununited ; surgery ; Humans ; Humeral Fractures ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Tibial Fractures ; surgery ; Treatment Outcome

OBJECTIVETo evaluate the clinical result of the treatment of old femoral neck fracture and nonunion with free vascularized fibular grafting.

METHODSFrom November 2000 to December 2005, 29 cases with old femoral neck fracture and nonunion had been treated by free vascularized fibular grafting with an average follow-up of 28.5 months.

RESULTSAll the fracture were healed without any severe complications. And the healing time was 4-6 months (5.6 months on average). During the follow-up, the hips of 28 cases got well-function, and the average Harris hip score was 88.2. One case came about with osteonecrosis of femoral head after one year and finally accepted THA after 2 years.

CONCLUSIONSThe free vascularized fibular grafting is a valuable procedure to treat old femoral neck fracture and nonunion.

Adult ; Bone Transplantation ; methods ; Female ; Femoral Neck Fractures ; surgery ; Fibula ; blood supply ; transplantation ; Follow-Up Studies ; Fractures, Ununited ; surgery ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Young Adult