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 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

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 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

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

Background: Cervical cancer is the sixth most common cancer in Chinese women. A standard treatment modality for cervical cancer is the combination of surgery, chemotherapy, external-beam radiotherapy and intracavitary brachytherapy. The aim of this study was to retrospectively assess the long-term treatment outcomes of patients with cervical cancer who were treated with californium-252 neutron brachytherapy combined with external-beam radiotherapy plus concurrent chemotherapy.Methods: We retrospectively analyzed the medical records of 150 patients with primary stages IB-IVB cervical cancer who received neutron brachytherapy combined with external-beam radiotherapy concurrently with cisplatin chemotherapy.All patients were followed up. Using an actuarial analysis, patient outcomes and treatment-related adverse effects were evaluated and compared.Results: The median overall survival (OS) was 33.2 months. The 3-year progression-free survival rates for patients with stages Ⅰ—Ⅱ, Ⅲ, and Ⅳ diseases were 81.0% (68/84), 65.0% (39/60), and 0% (0/6), respectively; the 3-year OS rates were 90.5% (76/84), 85.0% (51/60), and 16.7% (1/6), respectively. Vaginal bleeding was controlled within the median time of 4.0 days. One month after treatment, 97.3% of patients achieved short-term local control. The local recurrence rates for patients with stages Ⅰ—Ⅱ, Ⅲ, and Ⅳ disease were 4.8% (4/84), 11.7% (7/60), and 33.3% (2/6), respectively, and the occurrence rates of distant metastasis were 16.7% (14/84), 25.0% (15/60), and 100.0% (6/6), respectively. Cancer stage,tumor size, and lymph node metastasis were identified as prognostic risk factors, but only lymph node metastasis was found to be an independent prognostic factor. The most common adverse effects during treatment were grades 1 and 2 irradiation-related proctitis and radiocystitis.Conclusion: For patients with cervical cancer, neutron brachytherapy combined with external-beam radiotherapy plus concurrent chemotherapy produces a rapid response and greatly improves local control and long-term survival rates with tolerable adverse effects.

OBJECTIVETo explore the primary experimental methods to construct tissue engineering blood vessel.

METHODSUsing the collagen-chitosan to prefabricate the scaffolds with 3-dimensional structure, the proliferated human endothelial cells (ECs), smooth muscle cells (SMCs) and fibroblasts act as the seed cells. The cells were seeded to scaffolds in two-step method, and engineering tissue were matured by static culture or bioreactor culture. Extracellular matrix contents and the platelet aggregation were examined in engineering tissue, tissue engineering blood vessels were taken as patches to repair the man-made defaults on the rats aorta.

RESULTSThe proliferated human ECs, SMCs and fibroblast can hold activity and act as seed cells. The prefabricated scaffolds, with excellently cell and tissue biocompatibility, can facilitated cells adherence and upgrowth, the cells quantities and extracellular matrix contents in engineered tissue are time dependent increase (P < 0.05). Platelet aggregation tests confirm the tissue engineering blood vessel have some anti-coagulability. Using the engineering tissue patch to repair the default, 6 aortas in 8 animal were patency till 10 days post-operation.

CONCLUSIONSThe seeding cells can be seeded on the 3-dimensional collagen-chitosan scaffolds and matured, the tissue engineering blood vessel can be constructed primarily.

Animals ; Bioartificial Organs ; Biocompatible Materials ; Blood Vessel Prosthesis ; Cell Culture Techniques ; methods ; Cell Division ; Chitosan ; Collagen ; Endothelial Cells ; cytology ; Fibroblasts ; cytology ; Humans ; Myocytes, Smooth Muscle ; cytology ; Rats ; Rats, Sprague-Dawley ; Tissue Engineering ; methods