OBJECTIVETo study the medium and long term effects of perfusion of bone marrow stromal stem cells through optional artery for the treatment of non-traumatic femoral head avascular necrosis.

METHODSFrom January 2000 to December 2004,62 cases(78 hips) with non-traumatic femoral head necrosis accepted optional artery marrow stromal stem cells infusion treatment and had complete follow-up data, including 43 hips of 35 males and 35 hips of 27 females with an average age of 36.3 years old (22 to 54). According to preoperative imaging data, 16 hips were ARCO I stage, 52 hips were II stage, 10 hips were III a stage. Harris score was 64.94 +/- 8.12 preoperatively. Postoperative Harris score at the last follow-up, imaging changes,DSA vascular changes were analysis.

RESULTSThe patients were followed up for 9 to 13 years (means 11 years). By the end of the follow-up, a total of 18 hips got artificial joint replacement, 10 hips of preoperative ARCO I, II period got artificial hip joint replacement, 8 hips of IIIa period got hip artificial joint replacement. Harris score was 71.21 +/- 0.19 at the end of the follow-up, it was obviously enhanced compared with preoperative. DSA showed blood vessels of supply the femoral head increased thickening.

CONCLUSIONPerfusion of bone marrow stromal stem cells through optional artery can effective treat non-traumatic femoral head necrosis of ARCO I, II period, it can make the femoral circumflex artery and its branches increased thickening.

Adult ; Angiography, Digital Subtraction ; Arthroplasty, Replacement, Hip ; Female ; Femur Head Necrosis ; therapy ; Follow-Up Studies ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Middle Aged

Bone Marrow Cells ; metabolism ; pathology ; CD28 Antigens ; blood ; metabolism ; CD4 Antigens ; blood ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; Dendritic Cells, Follicular ; metabolism ; pathology ; Humans ; Immunoblastic Lymphadenopathy ; metabolism ; pathology ; Immunophenotyping ; methods ; Lymphoma, T-Cell ; metabolism ; pathology ; Neprilysin ; blood ; metabolism ; Receptors, Complement 3d ; blood ; metabolism ; fas Receptor ; blood ; metabolism

Catheterization ; adverse effects ; methods ; Catheterization, Central Venous ; adverse effects ; methods ; Female ; Humans ; Middle Aged ; Subclavian Artery ; surgery

Adolescent ; Adult ; Aged ; Brachial Plexus ; Female ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Nerve Block ; methods ; Shoulder Dislocation ; therapy

OBJECTIVETo explore mechanism of dermal toxicity of trichloroethylene(TCE).

METHODSNormal human keratinocytes (KC) were isolated from foreskins of healthy donors undergoing circumcision by two-step trypsin digestion and cultured in serum-free medium. Cells were treated with medium, 1% acetone (volume fraction) 0.125, 0.500 or 2.000 mmol/L TCE for different time (4, 8, 12 or 24) hours. After treatment, MTT assay and ATPase activity detected, inhibition ratio of mitochondrial enzyme was calculated according to optical density (A) value of MTT assay. Mitochondrial membrane potential (MMP) was detected by flow cytometry FCM after being stained with Rhodamine123 (Rh123). Morphological changes were also observed through transmission electron microscope (TEM).

RESULTSCellular viability and ATPase activity declined with dose of TCE, while inhibition ratio of mitochondrial enzyme increased with dose of TCE. FCM results showed that after treatment with 2.000 mmol/L TCE, fluorescence density of Rh123 decreased quickly from 18.73 +/- 0.45(0 h) to 8.20 +/- 0.66(8 h) (P < 0.01). After 8 h, fluorescence density maintained at the level equal to that of 8 h (fluorescence density of Rh123 were 8.20 +/- 0.36 and 8.20 +/- 0.40 for 12 and 24 h respectively, compared with that for 8 h group, P > 0.05). The results also showed that MMP diminished with dose of TCE. Under TEM, mitochondria in TCE-treated group appeared extensive swelling and vacuolar degeneration with less matrix and obscure or vanished mitochondria cristae but in control group, mitochondrial structure was integrated, with uniform matrix and visible mitochondria cristae.

CONCLUSIONSTCE could inhibit mitochondrial metabolic enzyme, reduce ATP production, diminish MMP, and destroy ultrastructure of mitochondria in KC, all these contributing to the cytotoxicity of TCE.

Cell Survival ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Humans ; Keratinocytes ; drug effects ; metabolism ; ultrastructure ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Microscopy, Electron, Transmission ; Mitochondria ; drug effects ; metabolism ; ultrastructure ; Trichloroethylene ; toxicity

Adolescent ; Adult ; Aged ; Ankle Joint ; pathology ; physiopathology ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Tuberculosis, Osteoarticular ; drug therapy ; pathology ; physiopathology ; surgery ; Young Adult

OBJECTIVETo observe the change of caspase-8, caspase-9 activity and apoptosis rates in the process of trichloroethylene-induced damage in keratinocytes, and explore the tentative mechanism of apoptosis.

METHODSHuman keratinocytes were exposed to 0.125, 0.250, 0.500, 1.000 and 2.000 mmol/L trichloroethylene for 4, 8, 12 and 24 h. The inhibitive groups were pretreated with 100 micromol/L Z-LEHD-FMK (a specific inhibitor of caspase-9) for 1 h, and were stimulated with 2.000 mmol/l TCE for 12 h. MTT assay was used to detect the viability of different cells; The activity of caspase were calculated according to spectrophotometry; Change of the apoptotic rates was assessed by flow cytometer (FCM) after double-stained with Annexin V-FITC and propidium iodide (PI).

RESULTS(1) The minimum effective concentration for cell viability reduction was 0.125 mmol/L at 12 h and the shortest time required to produce a change was 4 h at a concentration of 2.000 mmol/L (compared with control group, P < 0.01). Cell viability in all the groups markedly decreased from 12 h to 24 h (P < 0.05). (2) The activity of caspase-8 in the various dosage groups at different times had no statistical difference compared with the control group, P > 0.01. (3) At 8 h, 1.000 and 2.000 mmol/L TCE groups could significantly enhance caspase-9 activity (P < 0.05). The caspase-9 activity in all the groups showed differences and was significantly higher than those of control cells when time was over 12 h (P < 0.05). (4) After exposing to different dosages of TCE for 12 h, the rate of apoptosis rose to (80.43 +/- 4.21)% with the increase of dosage, compared with the control group, (9.40 +/- 2.98)%, which showed a dose-effect relationship. (5) The cells pre-treated with caspase-9 inhibitor resulted in a decrease in the caspase-9 activity and apoptosis rates (compared with 2.000 mmol/L TCE exposed group, P < 0.01). However, there was no statistical significance in comparison with the control group (P > 0.05).

CONCLUSIONCaspase-9 may be an important mediator of apoptosis in keratinocytes induced by trichloroethylene.

Apoptosis ; drug effects ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cells, Cultured ; Humans ; Keratinocytes ; drug effects ; enzymology ; pathology ; Trichloroethylene ; toxicity

BACKGROUNDRegulatory T cell populations, particularly CD4(+)CD25(+) T regulatory cells, have been implicated in the persistence of hepatitis B virus (HBV) infection. However, no clear relationship has been established between the frequency of CD4(+)CD25(+) T regulatory cells in the peripheral blood and either the disease phases in the natural history of chronic HBV infection or in the response to interferon-α therapy.

METHODSIn the present study, three different common markers of CD4(+)CD25(+) T regulatory cells were used to determine the numbers of T regulatory cells in healthy controls and in patients with chronic HBV infection.

RESULTSNo significant difference was found when samples were gated for CD25(hi) and CD25(+)FoxP3(+) T cells. A significant correlation was found between the number of CD4(+) Treg cells that gated with CD25(+)FoxP3(+) and CD25(+)CD127(low/-) in healthy controls and in patients with chronic hepatitis B (CHB) (r = 0.67, 0.59; P < 0.01). The percentages of Treg cells were (8.56 ± 2.01)% in asymptomatic carriers (Asc), (8.74 ± 3.04)% in inactive HBsAg carriers, (10.7 ± 2.93)% in CHB and (7.42 ± 1.28)% in healthy controls (F = 11.1, P < 0.001). The percentage of Treg cells in patients with CHB was higher than in asymptomatic HBV patients, inactive HBsAg carriers, or healthy controls (P < 0.01). The proportion of CD4(+)CD25(+)CD127(low/-) T cells in patients who responded to interferon-α was (11.9 ± 3.3)%, (9.1 ± 2.4)% and (9.0 ± 2.9)% at baseline, week 12 and week 24 after treatment, respectively (Z = 2.42, P < 0.05; Z = 2.67, P < 0.01).

CONCLUSIONSThese results suggest that the proportion of the CD4(+)CD25(+) regulatory T cells might be affected by the application of different markers in process to detect T regulatory cells. The frequency of Treg cells was increased in patients with CHB, which might be associated with the disease activity of these patients and contribute to prevention of extensive liver damage. A decline in Treg cells at week 12 of treatment might be associated with a better response to treatment with interferon-α.

Adult ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B Surface Antigens ; analysis ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Middle Aged ; T-Lymphocytes, Regulatory

OBJECTIVE: To evaluate the potential risk of porcine endogenous retrovirus (PERV) cross-species transmission xenotransplanted with microencapsulated neonatal pig islets (NPIs). METHODS: Ten dogs were randomly divided into an experiment group and a control group. The experiment group was transplanted with microencapsulated NPIs, and the control group was transplanted with non-microencapsulated NPIs. Glucose tolerance test (GTT) was performed to evaluate the function of microencapsulated NPIs after the transplantation; immunity histochemistry was used to detect the microencapsulated NPIs in the liver of dogs which had been transplanted after 28 days; PCR and RT-PCR were performed to detect PERV and pig mitochondrial (mt) DNA in the blood samples obtained from recipients at various time points after the transplantation. RESULTS: The level of serum special porcine C peptide increased significantly after the injection of glucose for 15 approximately 30 min in dogs which were transplanted with the micro-encapsulated NPIs over 2 weeks, while special porcine C peptide could not be detected in the control group. Immunity histochemistry showed that a few microencapsulated NPIs were still alive in the liver of the dog, and the liver was not damaged. PCR and RT-PCR showed that pig mt DNA and PERV could not be detected in the experiment group 1 approximately 28 days after the transplantation, while very weak expression of that in the control could be detected in the first 4 days and disappeared 10 days after the transplantation. CONCLUSION Microencapsulated NPIs can survive and have biological function in dogs. There is no evidence of PERV replication, suggesting that the xenotransplantation with microencapsulated NPIs can prevent PERV effectively, and may have great value.
Animals ; DNA, Mitochondrial ; isolation & purification ; Dogs ; Endogenous Retroviruses ; physiology ; Islets of Langerhans ; virology ; Islets of Langerhans Transplantation ; adverse effects ; Liver ; virology ; Swine ; Transplantation, Heterologous ; Virus Replication

OBJECTIVE: To investigate the change of nitric oxide (NO) level in local muscles induced by crushing hind-limbs in rats. METHODS: The rat experimental model of hind-limb crushing injury was established by crushing the hind limbs of rats with standard weight for 5 hours, thereafter releving the standard weight for another 5 hours. The rats were randomly divided into sham group, crushing group, crushing and injecting aminoguanidine (AG) group, crushing and injecting L-arginine (L-Arg) group. The NOS activity and NO level in local muscles and serum were spectrophotometrically measured, and iNOS and eNOS protein expressions in local muscles were examined by immunohistochemistry. The weight ratio of wet to dry (W/D) of local muscles was measured and the pathologic changes were observed. RESULTS: The crushing hind-limbs induced serious primary and secondary injuries of local muscles such as rupture and rhadomyolysis of skeletal muscular fibers, interstitial vascular congestion and edema, and marked increase in W/D. The expressions of eNOS and iNOS were upregulated in local muscle in crush group compared with sham group. The NOS activity and NO level in local muscles and serum significantly increased. There was positive relationship between NO level and W/D in local muscles. With the usage of AG and L-arg, the hind-limb injuries seemed alleviated and aggravated, respectively. CONCLUSION The crushing hind-limbs of rats elicited the upregulation of eNOS and iNOS protein expression, the enhancement of NOS activity and the excess production of NO, the latter of which was involved in the mediation of secondary pathological changes in local muscles.
Animals ; Disease Models, Animal ; Female ; Hindlimb/injuries* ; Immunohistochemistry ; Male ; Muscle, Skeletal/pathology* ; Nitric Oxide/blood* ; Nitric Oxide Synthase/metabolism* ; Rats ; Rats, Wistar ; Soft Tissue Injuries/pathology*