OBJECTIVETo investigate the effect of paraquat (PQ) on reactive oxygen species (ROS) and neutrophil apoptosis and its possible signal transduction pathways.

METHODSCultured neutrophils were treated with different concentrations of PQ for 6-24 h. The apoptosis rate of neutrophils and ROS content were determined by flow cytometry. The exoressions of nuclear factor kappa B (NF-κB) and Caspase 3 were detected by Western blot. These parameters were checked again after NF-κB and Caspase 3 antagonist were applied.

RESULTSPQ could boost ROS generation and depress neutrophil apoptosis significantly. At the same time PQ could enhance the expression of NF-κB and inhibit the expression of Caspase 3. These effects could be reversed by ROS inhibitor diphenyleneiodonium (DPI) and NF-κB inhibitor pyrrolidinedithiocarbamate (PDTC).

CONCLUSIONPQ is a potent inducer of ROS and can inhibit neutrophil apoptosis by activating NF-κB and surpressing Caspase 3 activity.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cells, Cultured ; NF-kappa B ; antagonists & inhibitors ; metabolism ; Neutrophils ; cytology ; drug effects ; Paraquat ; toxicity ; Pyrrolidines ; pharmacology ; Reactive Oxygen Species ; metabolism ; Signal Transduction ; Thiocarbamates ; pharmacology

Female ; Humans ; Infant, Newborn ; Maple Syrup Urine Disease ; diagnosis ; therapy

Abortion, Spontaneous ; diagnosis ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p57 ; metabolism ; Diagnosis, Differential ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Hydatidiform Mole ; diagnosis ; genetics ; metabolism ; Immunohistochemistry ; Pregnancy ; Uterine Neoplasms ; diagnosis ; genetics ; metabolism

OBJECTIVETo investigate luciferase gene expression and pathological changes of submandibular glands (SMG) of rats after adenovirus-mediated gene transfer.

METHODSAdenovirus-mediated luciferase gene (AdCMVLuc, 10(8) pfu in 50 microl) was injected in to SMG of forty wistar rats. The SMGs were harvested for gene expression measurement and pathological study after 3 days, 1,2,4 and 8 weeks.

RESULTSPeak expression was observed in three days following administration of the vector however, gene expression in submandibular glands decreased rapidly. the pathological changes induced by retrograde injection of AdCMVLuc included: after 3 days to one week, compression of acini, dilation of terminal ducts; after two weeks, slight atrophy of a part of acini, increase of iteracinar distance and focal lymphocyte infiltration in lobules and interlobular ducts; after 4 weeks, recovery evidence was found in acini; after 8 weeks, normal acini and ducts were found. The ultrastructural changes included: 3 days, much more rough endoplasmic reticulum was found both in acini and duct epithelial cell; a lot of mucus drops were found in acini; after 1 week, microvillus decreased in duct epithelial cells, mitochondria increased significantly in acini; intercellular space was enlarged.

CONCLUSIONSAdenovirus-mediated gene transfer can produce biological proteins and induce marked inflammatory changes in rat SMG. The ultrastructural changes suggest high protein synthesis activity in the acinar cells after gene transfer.

Adenoviridae ; enzymology ; genetics ; Animals ; Gene Expression ; Gene Transfer Techniques ; Luciferases ; genetics ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Submandibular Gland ; pathology ; virology

OBJECTIVETo evaluate an enzymatic method for determining serum beta-hydroxybutyrate (beta-HB) with the National Committee for Clinical Laboratory Standards (NCCLS) projects, and to discuss its clinical values in diabetic ketoacidosis (DKA).

METHODSThe precision, accuracy, specificity, linearity and interference of the enzymatic method were analyzed. This method was used to determine serum beta-HB in 60 cases of normals, 50 cases of diabetes, and 34 cases of DKA by autochemistry analyzer.

RESULTSEnzymatic beta-HB assay was precise (within-run CV, day-to-day CV, and total CV < 5%). The linearity studies showed the method was linear up to 4 mmol/L. Recovery rate was 98.5%-104.1%. Hemolysis (Hemoglobin up to 18.2 g/L), icteric samples with total bilirubin up to 224 mumol/L, and lipemia up to triglyceride concentration of 2.28 mmol/L did not interfere with the beta-HB results in this method. Serum beta-HB levels were significantly elevated in DKA patients compared with DM patients and controls (P < 0.01). Positive rate of serum beta-HB in DKA patients was significantly higher than that of urinary ketone (P < 0.05).

CONCLUSIONSEnzymatic method is convenient and reliable, allows full automation, and is rapid enough to be used for both routine and urgent determinations of serum beta-HB. It can be used in diagnosing and monitoring treatment of DKA.

3-Hydroxybutyric Acid ; blood ; Adolescent ; Adult ; Autoanalysis ; Diabetes Mellitus ; blood ; Diabetic Ketoacidosis ; blood ; Evaluation Studies as Topic ; Female ; Humans ; Male ; Middle Aged

OBJECTIVETo study the clinicopathologic features, diagnostic criteria, differential diagnosis and treatment options of ovarian steroid cell tumor, not otherwise specified (NOS).

METHODSLight microscopy and immunohistochemical study was carried out in 8 cases of ovarian steroid cell tumor, NOS. The literature was reviewed.

RESULTSThe 7 cases of benign ovarian steroid cell tumor, NOS were composed mainly of polygonal cells with granular eosinophilic cytoplasm and larger cells with vacuolated cytoplasm. They resembled the architecture of normal adrenal gland, with formation of cell nests and trabeculae. The single case of malignant ovarian steroid cell tumor had evidence of significant cellular pleomorphism, haemorrhage and coagulative tumor necrosis. The mitotic count measured about 7 per 10 high-power fields. Immunohistochemical study showed that the tumor cells expressed calretinin and alpha-inhibin. Differential diagnosis included oxyphilic granulosa cell tumor, thecoma, Sertoli cell tumor and clear cell carcinoma. The treatment options of benign ovarian steroid cell tumor, NOS was local excision or ipsilateral salpingo-oophorectomy, while the malignant counterpart should be treated with a combination of surgery and chemotherapy, including administration of GnRH agonist.

CONCLUSIONSOvarian steroid cell tumor, NOS, is the most common type of ovarian steroid cell tumors. Most of which are associated with a benign clinical outcome. Immunohistochemistry is an important adjunct for diagnosis. The treatment options of ovarian steroid cell tumor, NOS depend on its malignant potential.

Adolescent ; Adult ; Calbindin 2 ; Diagnosis, Differential ; Female ; Granulosa Cell Tumor ; pathology ; Humans ; Inhibins ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; surgery ; Ovariectomy ; methods ; Ovary ; pathology ; S100 Calcium Binding Protein G ; metabolism ; Sertoli Cell Tumor ; pathology ; Sex Cord-Gonadal Stromal Tumors ; metabolism ; pathology ; surgery ; Thecoma ; pathology ; Young Adult

The aim of this study was to investigate the expression of Th17 cells and regulatory T (Treg) cells in peripheral blood of patients with immune thrombocytopenia (ITP) and to clarify the role of the Th17/Treg cell ratio imbalance in pathogenesis of ITP. Patients were divided into the pre-treatment group (active group) (n = 38) and post-treatment group (remission group) according to the platelet count and curative effect. Post-treatment group was further divided into remission group (n = 24), partial remission group (n = 10), and non-remission group (n = 4). 30 healthy subjects were enrolled in control group. Flow cytometry was used to detect the percentages of peripheral blood Th17 cells and Treg cells in CD4(+) T cells from ITP patients and controls respectively. The results showed that the percentages of Th17 cells in active group and non-remission group were significantly higher than those in control group (p < 0.05). The percentages of Th17 cells in remission group, partial-remission group were also higher than those in control group, but there were no statistically significant differences between these groups. The percentage of Th17 cells in remission group was lower than that in active group, but there was also no statistically difference between two groups. The percentages of Treg cells in active group, partial-remission group and non-remission group significantly decreased, compared with in control group (p < 0.01). The percentage of Treg cells in remission group was lower than that in control group, but there was no statistically significant difference. The ratio of peripheral blood Th17/Treg cells in active group, partial-remission group and non-remission group was higher, as compared with in control group. The ratio of peripheral blood Th17/Treg cells in remission group was higher than that in control group, but there was no statistically difference between two groups. It is concluded the percentage of Th17 cells and the ratio of Th17/Treg cells are higher in active group. The percentage of Treg cells is low in active group, partial remission and non-remission groups. The imbalance of Th17/Treg ratio may play a critical role in ITP pathogenesis.
Adolescent ; Adult ; Aged ; Blood Cell Count ; Case-Control Studies ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Purpura, Thrombocytopenic ; blood ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology ; Young Adult

BACKGROUNDImiquimod is an imidazoquinoline, which class of compounds are known to have antiviral and antitumoural properties. In recent studies, it was shown that imiquimod modulates the T helper cell type Th1/Th2 response by inducing the production of Th1 cytokines like IFN-gamma, and by inhibiting the Th2 cytokines like interleukin (IL)-4. Several investigators have shown that T-bet and GATA-3 are master Th1 and Th2 regulatory transcription factors. This study investigated whether imiquimod treatment inhibited airway inflammation by modulating transcription factors T-bet and GATA-3.

METHODSThirty-six male SD rats were randomly divided into a control group, an asthmatic group, and an imiquimod group, which was exposed to an aerosol of 0.15% imiquimod. Twenty-four hours after the last ovalbumin (OVA) challenge, airway responsiveness was measured and changes in airway histology were observed. The concentrations of IL-4, IL-5 and IFN-gamma in bronchoalveolar lavage fluid (BALF) and serum were measured by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of IL-4, IL-5, IFN-gamma, T-bet and GATA-3 in lung and in CD4(+) T cells were determined by reverse transcription polymerase chain reaction (RT-PCR). The protein expressions of T-bet and GATA-3 were measured by Western blot.

RESULTSIt was demonstrated that imiquimod 1) attenuated OVA induced airway inflammation; 2) diminished the degree of airway hyperresponsiveness (AHR); 3) decreased the Th2 type cytokines and increased Th1 type cytokines mRNA and protein levels; 4) modulated the Th1/Th2 reaction by inhibiting GATA-3 production and increasing T-bet production.

CONCLUSIONImiquimod treatment inhibits OVA induced airway inflammation by modulating key master switches GATA-3 and T-bet that result in committing T helper cells to a Th1 phenotype.

Administration, Inhalation ; Aminoquinolines ; administration & dosage ; therapeutic use ; Animals ; Asthma ; drug therapy ; metabolism ; Bronchi ; pathology ; Bronchial Hyperreactivity ; drug therapy ; metabolism ; Cytokines ; biosynthesis ; Eosinophils ; physiology ; GATA3 Transcription Factor ; genetics ; Gene Expression Regulation ; drug effects ; Lung ; pathology ; Male ; Ovalbumin ; immunology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; T-Box Domain Proteins ; Transcription Factors ; genetics

Adult ; Bone Plates ; utilization ; Clavicle ; surgery ; Female ; Humans ; Internal Fixators ; trends ; Male ; Middle Aged ; Young Adult

OBJECTIVETo evaluate and analyze mechanism of Pizhen for the treatment of pain in patients with knee osteoarthritis.

METHODSEighty-five patients (85 knees) were divided into two groups according to random number table method. Among 35 patients in the experimental group, 6 patients were male and 29 patients were female, 1 patient lost follow-up, the average age was (56.65 +/- 4.89) years, and the patients were treated with Pizhen. Among 50 patients in the control group,there were 6 males and 44 females, the average age was (57.54 +/- 4.91) years, and the patients were treated with intra-articular injection with sodium hyaluronate. The VAS score and the data of algometer from the most obvious tenderness around the knee were used to evaluate the clinical effects.

RESULTSThere were significant differences of scores in the experimental group before and after treatment (P<0.01). There were significant differences of scores in the control group before and after treatment (P<0.01). After treatment, there were no statistical differences of VAS score and the data of algometer between experimental and control groups ( P>0.01). There was negative correlation between VAS score and the data of algometer.

CONCLUSIONThe Pizhen is an effective method to treat pain in patients with knee osteoarthritis by relaxing tendon of soft tissue around pain point of knee joint, and relaxing compress and stimulation of nerves.

Acupuncture Analgesia ; Aged ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; physiopathology ; therapy ; Pain Measurement