Objective To demonstrate the carbonic anhydrase Ⅲ (CAⅢ) for 25 000 protein decreased in skeletal muscle of myasthenia gravis (MG). Methods The protein molecular properties responsible to antibodies against 25 000 protein and CAⅢ were analyzed by a combination method of two-dimensional electrophoresis and immuno-Western blot. Competitive binding reactions of the antibodies to the purified 25 000 protein and muscular homogenate were observed by using immuno-Dot blot and immuno-Western blot, respectively. The expression of CAⅢ from normal and MG muscles was detected by immuno-Western blot. Results Combination analysis of two-dimensional electrophoresis and immuno-Western blot showed that the protein of immunological responsible to antibodies against 25 000 protein and CAⅢ had an identical molecular mass and isoelectric point. Competitive binding reactions proved that 25 000 protein and CAⅢ were the same substance, either by immuno-Dot blot or by immuno-Western blot. In addition, a much similar result was obtained when the levels of 25 000 protein from normal and MG muscles were detected by antibodies against 25 000 protein and (CAⅢ) by immuno-Western blot. Conclusion 25 000 protein decreased in the MG skeletal muscle was proved to be just a known protein CAⅢ, which made a basis for further exploring the relationship of CAⅢ deficiency and MG pathogenesis.

OBJECTIVETo observe the clinical efficacy of treatment for clearing heat, detoxicating and nourishing Yin (abbr. as CHM) on glucocorticosteroid (GCS) induced facial dermatitis, and its effect in repairing skin barrier function.

METHODSOne hundred and fifteen patients were randomly assigned into two groups, 68 in the treated group treated with CHM and 47 in the control group treated by oral administration of loratadine 10 mg once a day and ketotifen 1 mg once every night. The volume of transepidermal water loss (TEWL) of patients was measured before and after treatment.

RESULTSThe effective rate was 77.9% (53/68) and 14.9% (7/47) in the treated group and the control group respectively, showing significant difference between the two groups, and it was better in the treated group than that in the control group (P < 0.01). The decrease of TEWL in the treated group was also superior to that in the control group (P < 0.01).

CONCLUSIONChinese herbal treatment for clearing heat, detoxicating and nourishing Yin has significant clinical efficiency on GCS induced facial dermatitis and in repairing the skin barrier function.

Adolescent ; Adult ; Aged ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Facial Dermatoses ; chemically induced ; drug therapy ; Female ; Glucocorticoids ; adverse effects ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy ; Skin Absorption ; Skin Physiological Phenomena ; drug effects ; Yin Deficiency ; drug therapy

An alkaline catalase has been purified and characterized from a slightly halophilic and alkaliphilic bacterium Bacillus sp. F26. The purification was performed with a four step procedure consisting of ammonium sulfate precipitation, ion exchange, gel filtration and hydrophobic interaction chromatography, and finally achieved a 58.5-fold-purifying over the crude extract. The purified catalase was composed of two identical subunits with a native molecular mass of 140 kD. The native enzyme showed the typical Soret band appearing at 408 nm. The pyridine hemochrome spectrum indicated the presence of protoheme IX as the prosthetic group. The apparent Km value for enzyme activity on H2O2 was calculated to be 32.5 mmol/L. The activity of this catalase was not reduced by dithionite but was strongly inhibited by cyanide, azide, and 3-amino-1,2,4-triazole (the specific inhibitor of monofunctional catalase). No peroxidase activity of this enzyme was detected when using o-dianisidine, diaminobenzidine (DAB) and p-phenylenediamine as electron donor. Moreover, the N-terminal sequence of this catalase exhibited substantial similarity to the monofunctional catalase subgroup rather than catalase-peroxidase or Mn-catalase one. Therefore, we characterize the purified catalase as a monofunctional catalase. Besides, this monofunctional catalase was thermosensitive and its activity exhibited pH-independent over pH 5-9 but showed a sharp maximum at pH 11. An activity half-life of approximately 49 h was measured when the enzyme was incubated at 20 degrees C and pH 11. To our knowledge, pH 11 is the most alkaline condition for optimum catalysis and enzyme stability among the catalases reported up to now. Furthermore, this monofunctional catalase also showed excellent halo-alkali-stability with a half-life of approximately 90 h at 0.5 mol/L NaCl and pH 10.5. On the other hand, so far as we know, the characterized catalase is the first dimeric monofunctional catalase from alkaliphiles and is also the first monofunctional catalase derived from a natural soda lake, which could partially reflect the oxidative stress response in the corresponding environment.
Bacillus ; enzymology ; Bacterial Proteins ; chemistry ; isolation & purification ; Catalase ; chemistry ; isolation & purification ; Enzyme Stability ; Hydrogen-Ion Concentration ; Temperature

AIMTo investigate the effects of NPPB, a chloride channel blocker, on proliferation of mesangial cells.

METHODSCell proliferation was determined by measuring cell number and 3H-thymidine incorporation. The LDH activity released from these cells was measured as evaluation of cell viability. The phase of cell cycle was detected by flow cytometry.

RESULTSCell proliferation assays showed that treatment with both NPPB (50 and 25 micromol x L(-1)) and in hypertonic media (100% increased osmolarity with D-mannitol ) significantly reduced the number of human MC and 3H-thymidine incorporation in a dose-dependent manner. But the LDH activity was not significantly altered in the treatment with 50 micromol x L(-1) NPPB. Flow cytometry experiments showed that 50 and 25 micromol x L(-1) NPPB arrested (84.2 +/- 2.4) % and (80.8 +/- 2.9) % of cells at G0/G1 stage, versus (70.5 +/- 1.4) % of control cells. Conclusion NPPB suppresses cell proliferation and produces growth arrest at G0/G1 phase in human MC by a mechanism probably associated with changes in cell volume.

Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chloride Channels ; antagonists & inhibitors ; Dose-Response Relationship, Drug ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Mesangial Cells ; cytology ; metabolism ; Nitrobenzoates ; administration & dosage ; pharmacology

BACKGROUNDCell-based vascular therapies of endothelial progenitor cells (EPCs) mediated neovascularization is still a novel but promising approach for the treatment of ischemic disease. The present study was designed to investigate the therapeutic potentials of human umbilical cord blood-derived EPCs (hUCB-EPCs) in rat with acute myocardial infarction.

METHODSHuman umbilical cord blood (hUCB) mononuclear cells were isolated using density gradient centrifugation from the fresh human umbilical cord in healthy delivery woman, and cultured in M199 medium for 7 days. The EPCs were identified by double-positive staining with 1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethylindocarbocyanine percholorate-labeled acetylated low-density lipoprotein (Dil-Ac-LDL) and fluorescein isothiocyanate-conjugated Ulex europaeus lectin (FITC-UEA-l). The rat acute myocardial infarction model was established by the ligation of the left anterior descending artery. The hUCB-EPCs were intramyocardially injected into the peri-infarct area. Four weeks later, left ventricular function was assessed by a pressure-volume catheter. The average capillary density (CAD) was evaluated by anti-VIII immunohistochemistry staining to reflect the development of neovascularization at the peri-infarct area. The graft cells were identified by double immunofluorescence staining with human nuclear antigen (HNA) and CD31 antibody, representing human origin of EPCs and vascular endothelium, respectively. Expressions of cytokines, proliferating cell nuclear angigen (PCNA), platelet endothelial cell adhesion molecule (PECAM) and vascular endothelial growth factor (VEGF) were detected to investigate the underlying mechanisms of cell differentiation and revascularization.

RESULTSThe donor EPCs were detectable and integrated into the host myocardium as confirmed by double-positive immunofluorescence staining with HNA and CD31. And the anti-VIII staining demonstrated a higher degree of microvessel formation in EPCs transplanted rats, associated with a significant improvement of global heart function in terms of the increase of left ventricular end-systolic pressure (LVESP), +dp/dtmax and -dp/dtmax as well as the decrease of LVEDP in rats with EPCs therapy comparing to the control rats (P < 0.05). Moreover, the expression of the rat PCNA mRNA and PECAM were both enhanced in the EPCs group compared with that of the control group.

CONCLUSIONSThe human umbilical cord blood-derived EPCs could incorporate into new-born capillaries in rat myocardium, induce revascularization and improve the proliferation activity in the peri-infarct area, resulting in the improvement of global heart function. This may indicate a promising stem cell resource in cell-based therapy for ischaemic diseases.

Animals ; Cells, Cultured ; Cytokines ; metabolism ; Endothelial Cells ; cytology ; physiology ; Endothelium, Vascular ; Fluorescent Antibody Technique ; Humans ; Male ; Myocardial Infarction ; metabolism ; therapy ; Neovascularization, Physiologic ; physiology ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; Rats ; Rats, Wistar ; Stem Cell Transplantation ; Stem Cells ; cytology ; Umbilical Cord ; cytology ; Vascular Endothelial Growth Factor A ; metabolism

Objective To report the experience of the application of microsurgery in joint replace- ment.Methods There were 22 cases,10 cases with segmental acetabular defects treated with the pedicle sartorius muscle iliac bone grafting,5 cases with vascular repair following major vascular injury of extremity during operation,6 cases with neural repair following neural injury during operation,1 case with serious injury reconstruction by elbow joint replacement and free flap.Results The operations succeeded in 22 cases without any postoperative infection.The mean follow-up was 40.1 months (3-72 months) in 22 cases,in which the joint function improved and the operative result was satisfactory with no joint pain.Conclusion Microsurgical technique can reconstruct bone and tissue defect effectively in joint replacement.

Objective To retrospectively study revisions for infected total hip replacements in 30 cases and discuss the bacteriological characters of the infected total hip replacements,difficulties and strategies in the revision.Methods Thirty revisions of infected total hip replacements were reviewed retrospectively.There were 12 males and 18 females,with mean age of 62.5 years(31-86 years)at revi- sion surgery.Infection was presented one month to four years(mean seven months)after THA operation. The diseases for initial operation included femoral neck fractures in 12 cases,femoral head necrosis in 11,hip osteoarthritis in five and rheumatoid arthritis in two.Twelve eases were treated by one-stage revi- sion and 18 by two-stage revision.Results Before the revision operation,the hip infection were diag- nosed by bacterial culture in 18 cases including five with Staphylococcus epidermidis,four with Staphylo- coccus aureus and nine with other bacteria.Bacteria growth appeared on the specimens from 23 hip joints during the revision surgery but not on the specimens from seven hip joints.Of 12 one-stage revisions,10 cases were followed for mean 16 months,which showed infection recurrence in two eases.Of 18 two-stage revisions,13 cases were followed for mean 20 months,which showed one case with infection recurrence. The mean Harris hip score was improved from preoperative 44 to 84 at follow-up.Conclusions 1) The main bacteria in the infected hip are antibiotic resistant Staphylococcus.2)Because the revision op- eration is difficult,careful preparation before revision is important for success.The fresh surgeon should not attempt.3)The revision strategies should vary according to specific status of the cases.The infection recurrence rate is lower when using a two-stage revision strategy.4)Application of antibiotic bone cement can help improve treatment effect and facilitate functional recovery of the joints.5)The scientific rehabil- itation after operation is very important to functional recovery.

OBJECTIVE: To study the value of fractional anisotropy (FA) of regions of interest (ROI) on magnetic resonance diffusion tensor imaging (DTI) in bilirubin-induced neurological dysfunction in neonates. METHODS: A total of 91 neonates with hyperbilirubinemia who were hospitalized from January 2017 to January 2018 were enrolled. According to the peak level of total serum bilirubin, they were divided into three groups: mild/moderate increase (n=45), severe increase (n=35), and extremely severe increase (n=11). According to the presence or absence of abnormal neurological manifestations, they were divided into two groups: neurological dysfunction (n=20) and non-neurological dysfunction (n=71). Ten healthy full-term infants were enrolled as the control group. Head DTI was performed for all neonates to measure the FA values of the bilateral globus pallidus, the anterior limb of the internal capsule, the posterior limb of the internal capsule, and the cerebellar dentate nucleus. RESULTS: The extremely severe increase group had significantly lower FA values of the globus pallidus than the control, mild/moderate increase, and severe increase groups (P<0.05). The severe increase group had significantly lower FA values of the globus pallidus than the control group (P<0.05). The extremely severe increase group had significantly lower FA values of the posterior limb of the internal capsule than the control, mild/moderate increase, and severe increase groups (P<0.05). The neurological dysfunction group had significantly lower FA values of the globus pallidus and the posterior limb of the internal capsule than the non-neurological dysfunction group (P<0.05). CONCLUSIONS Serum bilirubin level combined with the changes in the DTI FA values of the globus pallidus and the posterior limb of the internal capsule can be used to predict the injury of cerebral nuclei and white matter fibers.
Anisotropy ; Bilirubin ; Brain ; Diffusion Magnetic Resonance Imaging ; Diffusion Tensor Imaging ; Humans ; Infant, Newborn ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; White Matter

OBJECTIVETo analyze retrospectively the surgical treatment of 21 cases of infected arterial injuries of the major limb hospitalized from 1989 to 2003 in our department.

METHODSAfter a radical debridement and drainage, an autologous vessel bypass reconstruction, simple vessel ligation or amputation were carried out respectively according to each patient's condition. In order to make a better milieu for the grafts, local musculo-cutaneous flaps were translocated to cover the wounds in 9 cases.

RESULTS1 case (4.8%) died during the treatment. 5 cases (5/21, 23.8%) underwent amputations. Blood supply reconstruction was successful in 15 cases (15/21, 71.4%). 93.3% (14/15) of the cases were successful in getting an unobstructed vessel at early stage. Thrombosis occurred in 1 case (1/15, 6.7%) and a sound result was obtained after a second operation.

CONCLUSIONSRadical debridement combined with bypass vessel reconstruction with autologous vein and with musculo-cutaneous flap translocation is an effective treatment for infective limb vascular injuries.

Objective To explore the surgical effect to traumatic tentorial herniation with bilater- al mydriasis.Methods The patients were divided into three groups,ie,epidural hematoma group,a- cute diffuse brain swelling group and cerebral contusion and/or subdural hematoma group,to perform clinical outcome analysis.Half year after operation,the neurological outcome was scored according to the Glasgow Outcome Scale.Results Of all,there were three cases with good recovery,10 with moderate disability,nine with severe disability and 10 with vegetative survival but 35 deaths.The outcome was the best in epidural hemotoma group but the poorest in acute diffuse brain swelling group.Conclusions The operative effect of traumatic cerebral herniation with bilateral mydriasis is related with the type of orig- inal injury that is important for selection of operation.Patients with cerebral herniation caused by epidural hematoma should receive immediate operation that will induce better outcome.The operation is not vital for those with cerebral herniation caused by acute diffuse brain swelling.Emergent surgery can save lives of some patients with cerebral contusion and/or subdural hematoma.Rapid diagnosis,correct operation and perioperative treatment may ensure the success of surgery.