BACKGROUND: Ischemic preconditioning (IPC) can induce endogenous protection mechanism, which effectively prevent ischemia/reperfusion injury following organ transplantation. Cold and warm ischemia may induce ischemia/reperfusion injury of pancreas transplantation, and apoptosis of pancreatic acinar cells is one of the important reasons of pancreas graft functional defect after transplantation. Mitochondrial DNA has repair system, and its balance with mitochondrial DNA injury influences disease occurrence and outcome.OBJECTIVE: To observe the effect of IPC on apoptosis of transplanted pancreatic acinar cells, and the possible role of reactive oxygen (ROS) and mitochondrial DNA repair enzyme.METHODS: A total of 50 health, male, Sprague-Dawley rats were randomly divided into three groups: sham operated (n = 10), donors (n = 20) and recipients (n = 20). The recipients were randomly divided into ischemia/reperfusion group (IR, n = 10) and IPC group (n = 10). The sham operated group was subjected to abdominal open and close operation. IR group and IPC group received establishment of diabetic model by streptozotocin injection. IR rats received whole pancreatic-duodenal transplantation alone. IPC rats received whole pancreatic-duodenal transplantation exposed ischemic preconditioning with 5 minutes ischemia and 5 minutes reperfusion twice. All grafts were keep with warm ischemia time 15 minutes and cold ischemia (in 4 ℃ UW preservation solution) time 180 minutes. Twelve hours after reperfusion, serum amylase, blood glucose, Caspase-3, -9 activity were detected. Pancreatic acinar cell apoptosis was measured by flow cytometry. Mitochondrial cross-membrane potential (Δψ) was measured by monitoring the fluorescence spectrum of rhodamine 123. Mitochondrial H2O2 generation was determined using dichlorofluorescein as a probe. 8-oxodG in mitochondrial DNA (mtDNA) was measured with HPLC system. Release of cytochrome C, phosphorylation of Akt and mitochondrial OGG1 protein expression were determined by Western-blotting. RESULTS AND CONCLUSION: The ischemia preconditioning can relieve the pancreatic acinar cell apoptosis in pancreas graft and relieve IR injury by decreasing mitochondrial oxidative stress, mtDNA injury, and increasing phosphorylation of Akt and mitochondrial OGG1 expression.

OBJECTIVETo observe the effect of Ningdong Granule (NG) on serum levels of interleukin-12 (IL-12) and tumor necrosis factor-alpha (TNF-alpha) of children patients with Tourette's syndrome (TS).

METHODSTotally 90 TS children patients were randomly assigned to the NG group, the NG + Tiapride group (abbreviated as the combined treatment group), and the Tiapride group, 30 in each group. Besides,another 30 healthy children were recruited as the healthy control group. Patients in the NG group were treated with NG (consisting of all gastrodia rhizome, Codonopsis pilosula, Ophiopogon japonicus, white peony root, Rhinocerotidae, oyster, earthworm, licorice root, etc.), one dose daily, administered by dissolving it in boiled water, taken in two portions in the morning and in the evening respectively. Patients in the Tiapride group took Tiapride Tablet, 50 -100 mg each time, twice daily. The dosage was adjusted according to individual difference and changes of pathogenic conditions. The maximal dosage was 300 mg per day. Those in the combined treatment group were treated with equal dose of NG and Tiapride Tablet in combination. The treatment course was 3 months for all. Changes of pathogenic condition before and after treatment were assessed by Yale global tic severity scale (YGTSS). Serum levels of IL-12 and TNF-alpha were detected by enzyme-labeled immunosorbent assay (ELISA) before and after treatment.

RESULTS(1) The total effective rate of the NG group, the combined treatment group, and the Tiapride group was 79.3%, 83.3%, and 67.9%, respectively. It was the lowest in the Tiapride group (P < 0.05). It was significantly higher in the combined treatment group than in the NG group (P < 0.05). (2) The post-treatment YGTSS score was obviously lower in each group after treatment than before treatment (P < 0.05). The posttreatment YGTSS score was obviously lower in the NG group and the combined treatment group than in the Tiapride group (P < 0.05), but with no statistical difference between the fromer two groups (P > 0.05).(3) Compared with the healthy control group before treatment, serum levels of IL-12 and TNF-alpha (pg/mL) were 124.95 +/- 22.78 and 209.52 +/- 21.69 in the NG group, 126.14 +/- 25.65 and 208.97 +/- 22.46 in the combined treatment group, 123.00 +/- 24.26 and 205.10 +/- 26.16 in the Tiapride group, being higher than those in the healthy control group (64.56 +/- 27.59 and 78.13 +/- 33.42; P < 0.05). After treatment, serum levels of of IL-12 and TNF-alpha were 104.67 +/- 16.84 and 183.01 +/- 24.95 in the NG group, 109.04 +/- 16.81 and 179.87 +/- 23.45 in the combined treatment group, significantly lower than before treatment (P < 0.05). But there was no statistical difference in serum levels of IL-12 or TNF-alpha in the Tiapride group between before treatment (123.00 +/- 24.26 and 205.10 +/- 26.16) and after treatment (117.75 +/- 16.79 and 199.76 +/- 33.21; P > 0.05).

CONCLUSIONNG could modulate abnormal serum levels of IL-12 and TNF-alpha in TS children patients, which might be one of its pharmacodynamic mechanisms for treating TS.

Adolescent ; Child ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interleukin-12 ; blood ; Male ; Phytotherapy ; Tourette Syndrome ; blood ; drug therapy ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo determine the main factors which affect the percutaneous penetration of artesunate and provide efficient data for the artesunate transdermal delivery system.

METHODTransdermal speed constant and accumulative amount of 12 hours were used for the estimations of various reservior vehicles, and the supplement orthodox design was used to study the effect of pH, various proportion of IPA/Water/IPM, and drug concentration.

RESULTDrug concentration and pH were the main factors which affected the percutaneous penetration of artesunate.

CONCLUSIONThe suitable reservior vehicle can prompt the percutaneous penetration of artesunate, and artesunate TTS will be made with further studies.

2-Propanol ; pharmacology ; Administration, Cutaneous ; Animals ; Antimalarials ; administration & dosage ; pharmacokinetics ; Artemisinins ; administration & dosage ; pharmacokinetics ; Drug Carriers ; Drug Delivery Systems ; Hydrogen-Ion Concentration ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Sesquiterpenes ; administration & dosage ; pharmacokinetics ; Skin Absorption ; drug effects

OBJECTIVETo study the effect of internal fixation with absorbable pins on treatment of displaced radial head fractures.

METHODSFrom May 1999 to May 2004, 16 patients with displaced radial head fractures (Mason types II and III) were treated with internal fixation by absorbable pins. The duration of follow-up averaged 22.6 months (12-58 months). The outcome was assessed on the basis of elbow motion, radiographic findings and the functional rating score delineated by Broberg and Morrey.

RESULTSAll fractures healed within 10 months without avascular necrosis of radial head. The mean elbow flexion loss was 15 degrees (0 degrees-35 degrees), and pronation and supination decreased by 10 degrees (0 degrees-30 degrees) on average compared with those of the contralateral elbow. Five patients had an excellent result, 6 a good result, and 3 a fair result according to the criteria of Borberg and Morrey.

CONCLUSIONSInternal fixation with absorbable pins is an effective method in treating displaced radial head fractures. It can maintain the biomechanical stability of forearm, improve the elbow function and avoid second operation.

Adult ; Biomechanical Phenomena ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Prostheses and Implants ; Prosthesis Design ; Radius ; surgery ; Radius Fractures ; physiopathology ; surgery

Objective To observe the effects of ultra-wideband (UWB) electromagnetic irradiation on the ultrastructure of pituitary or testis and the serum sex hormones level of in rats. Methods SD male rats were divided randomly into control group and irradiation groups exposed to 3×105 pulses UWB irradiation for 6,12, 24, 48 h. After exposure, the ultrastructure changes of pituitary and testis were observed by electron microscope. Serum testosterone (T) , luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by radioimmunoassay. Results The rat pituitary and testis were injured at the different time after exposure to UWB electromagnetic irradiation, but the damage was serious at the 24 h after exposure. In the basophilic cell of the pituitary, there were the vacuoles and lipid droplets, dilated endoplasmic reticulum (ER),exuded lymphocytes and gathered chromatin in the cell border. In the testis tissue, there were the dilated ER and gathered chromatin in the cell border of the spermatogonium, interstitial cell and sustentacular cell, some mitochondria became vacuolar and swollen in the capillary endotheliocytes and exuded lymphocytes. Serum Tlevels of the irradiation groups at 24 and 48 h were ( 1209.7±115.7 ), ( 1340.5±331.1 ) μg/L, which were significantly lower than those[(2721.8± 178.9) and ( 2820.9±321.4 ) μg/L]of control groups at 24 and 48 h (P＜0.05). There was no significant difference for LH and FSH between exposure groups and control groups. Conclusions UWB electromagnetic radiation could induce some changes in the ultrastructure of rat pituitary and testis and serum T level, which may induce the change of reproductive system.

Objective To investigate the in vivo gene expression of adenovirus-mediated human tissue factor pathway inhibitor (hTFPI) and its inhibition effects on intimal proliferation in rabbit carotid arteries after ballon injury. Methods Rabbits underwent carotid artery balloon injuries were treated with Ad-TFPI (n=25), Ad-LacZ (n=25) or PBS (n=10), respectively. Sham operated rabbits (n=10) serve as normal controls. The expressions of human TFPI at mRNA and protein levels were detected by RT-PCR and ELISA respectively on the 3rd, 7th, 10th, 14th, 28th day after operation. Intimal proliferation was detected by angiograms and morphometric analysis. Results TFPI mRNA and protein expressions were detected at 3 days and peaked at the 10th and 14th day after TFPI gene transfer. The expressions were still detectable on the 28th day. There was no TFPI expression in Ad-LacZ group. The carotid angiogram results indicated that the minimal lumen diameter in TFPI group was significantly larger and the lumina stenosis percentage was significantly lower in TFPI group compared those in Ad-LacZ and PBS groups (all P < 0.05). The morphometric analysis showed that the intimal area, the ratio of the intimal/media area, the lumina stenosis percentage in TFPI group were all significantly reduced compared with those in Ad-LacZ and PBS groups (all P<0.01).Conclusions The TFPI gene could be effectively transferred by adenovirus vector to injured carotid arteries in rabbits.

Objective To observe the effects of ultra-wideband (UWB) electromagnetic irradiation on the ultrastructure of pituitary or testis and the serum sex hormones level of in rats. Methods SD male rats were divided randomly into control group and irradiation groups exposed to 3×105 pulses UWB irradiation for 6,12, 24, 48 h. After exposure, the ultrastructure changes of pituitary and testis were observed by electron microscope. Serum testosterone (T) , luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by radioimmunoassay. Results The rat pituitary and testis were injured at the different time after exposure to UWB electromagnetic irradiation, but the damage was serious at the 24 h after exposure. In the basophilic cell of the pituitary, there were the vacuoles and lipid droplets, dilated endoplasmic reticulum (ER),exuded lymphocytes and gathered chromatin in the cell border. In the testis tissue, there were the dilated ER and gathered chromatin in the cell border of the spermatogonium, interstitial cell and sustentacular cell, some mitochondria became vacuolar and swollen in the capillary endotheliocytes and exuded lymphocytes. Serum Tlevels of the irradiation groups at 24 and 48 h were ( 1209.7±115.7 ), ( 1340.5±331.1 ) μg/L, which were significantly lower than those[(2721.8± 178.9) and ( 2820.9±321.4 ) μg/L]of control groups at 24 and 48 h (P＜0.05). There was no significant difference for LH and FSH between exposure groups and control groups. Conclusions UWB electromagnetic radiation could induce some changes in the ultrastructure of rat pituitary and testis and serum T level, which may induce the change of reproductive system.

OBJECTIVEThe pathophysiology of beta-thalassemia is the imbalance of the alpha and non-alpha globin chain which leads to a series of clinical symptoms of hemolytic anemia. Scientists continuously try to explore gene-activated drugs to increase the level of non-alpha globin chain or decrease the level of alpha globin chain in the treatment of beta-thalassemia. To probe into the effects on globin-gene expression of meisoindigo (Me) in cultured erythroid cells derived from peripheral blood, so as to provide the theoretical basis for applying Me in the treatment of beta-thalassemia.

METHODSBy using the two-step liquid culture of erythroid progenitor cells and reverse transcription polymerase chain reaction (RT-PCR), and by using alpha mRNA as an inner control, the level of gamma mRNA and beta mRNA in cultured erythroid cells derived from peripheral blood of 11 patients with severe beta-thalassemia and 6 normal volunteers were measured under the effect of different concentration (2.5 micro mol/L, 5 micro mol/L and 10 micro mol/L) of Me.

RESULTS(1) No statistic significance was found in the ratio of beta/alpha mRNA by Me in cultured cells from both normal individuals and beta-thalassemia. (2) Me can significantly increase the ratio of gamma/alpha mRNA and (beta + gamma)/alpha mRNA (that is non-alpha/alpha mRNA) in cultured cells from normal individuals and beta-thalassemia. The ratio of gamma/alpha mRNA was increased 0.31 - 0.45 times and the ratio of non-alpha mRNA/alpha mRNA increased 0.21 - 0.32 times in Me induced cells from normal individuals. No significant result was observed among the different concentrations of Me (2.5 micro mol/L, 5 micro mol/L and 10 micro mol/L) in normal individuals. With the increasing of Me concentrations, the ratios of gamma/alpha mRNA and alpha/alpha mRNA were increased in cultured cells from beta-thalassemia. The ratio of gamma/alpha mRNA was increased 0.33 - 1.17 times and the ratio of non-alpha/alpha mRNA increased 0.25 - 0.89 times in Me induced cells from beta-thalassemia. There was no significant difference between the concentrations of 2.5 micro mol/L and 5 micro mol/L concentration in beta-thalassemia. However, there was significant difference between the concentrations of 10 micro mol/L and the concentrations of 2.5 micro mol/L and 5 micro mol/L in beta-thalassemia. (3) The increase of the ratio of gamma/alpha mRNA and non-alpha/alpha mRNA in beta-thalassemia was higher than that in normal individual with induction by Me with a higher concentration (10 micro mol/L).

CONCLUSIONMe can raise the ratio of gamma/alpha mRNA and non-alpha/alpha mRNA in cultured erythroid cells derived from peripheral blood of both normal individual and beta-thalassemia in the level of transcription, which can improve the imbalance of the alpha and non-alpha globin chain. So Me has a latent value in the therapy of beta-thalassemia.

Cells, Cultured ; Child ; Child, Preschool ; Erythroid Precursor Cells ; drug effects ; metabolism ; Female ; Gene Expression ; drug effects ; Globins ; genetics ; Humans ; Indoles ; pharmacology ; Infant ; Male ; RNA, Messenger ; drug effects ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore a new method and estimate its outcome to repair deep heel tissue injuries with local tissue flap.

METHODSAt lower cnemis, a fat fascial flap combined with perineal tendofascial flap were designed and elevated between the lateral malleolars blank, the largest area is 6 cm x 13 cm, which were transplanted to cover naked calcaneal tendon and calcaneal bone, full-thickness free skin graft or middle-thickness free skin graft were grafted on flap.

RESULTS6 patients suffered from heel tissue defects had been treated from January 2004 to November 2005. The smallest area was 3.5 cm x 4.0 cm and the largest was 4 cm x 6 cm. All fascial flaps and free skin grafts were successfully survived after operations. All patients were followed up 3-23 months, the operation areas were healed, the functions and configurations were satisfied.

CONCLUSIONSPerineal tendofascial flap conjoined with adipofasical flap and free skin graft are used to repair heel deep tissue defects, which is a handy, dependable way for repairing of the small tissue defects of the lower third of leg and heel.

Achilles Tendon ; injuries ; surgery ; Adult ; Fascia ; transplantation ; Female ; Fibula ; surgery ; Heel ; injuries ; surgery ; Humans ; Male ; Middle Aged ; Skin Transplantation ; Soft Tissue Injuries ; surgery ; Surgical Flaps

OBJECTIVETo provide an cell model of immortalized lymphoblstoid B-cell lines for studying the biological characteristics of full-length hepatitis B virus (HBV) genome carrying the hot-spot mutations V60, G87, and L97.

METHODSV60, G87, and L97 mutation points were introduced into HBV p3.8 II plasmid containing 1.2 copy of HBV genome by means of site-directed mutagenesis. The HBV genome was amplified by PCR from p3.8 II and p3.8 II-V60, G87, L97 plasmid, and the PCR product was inserted into EBO-plpp eukaryotic expression vector. The recombinant vectors and the EBO-plpp vector were transfected into immortalized human lymphoblasts with lipofectamine 2000 and selected with hygromycin. Steady expression of the target genes was determined by RT-PCR, Western blotting and microparticle enzyme immunoassay.

RESULTSDNA sequence analysis indicated that the desired mutation was introduced into wild-type HBV DNA. HBsAg, HBeAg and HBcAg could be detected in EBO-HBV-transfected cell lysate or culture supernatant.

CONCLUSIONTransfectants that stably express HBV mutant antigen may provide a cell model to study the biological characteristics of HBV carrying hot-spot mutation in vitro.

B-Lymphocytes ; cytology ; virology ; Base Sequence ; Blotting, Western ; Cell Line, Transformed ; Cell Transformation, Viral ; DNA, Viral ; genetics ; Eukaryotic Cells ; metabolism ; Gene Expression Regulation, Viral ; Genetic Vectors ; Genome, Viral ; genetics ; Hepatitis B Core Antigens ; genetics ; metabolism ; Hepatitis B virus ; genetics ; metabolism ; Humans ; Molecular Sequence Data ; Point Mutation ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection