Crystallization ; Eosinophils ; enzymology ; Fascioliasis ; pathology ; Granuloma ; pathology ; Humans ; Liver ; pathology ; ultrastructure ; Lung ; pathology ; ultrastructure ; Lung Diseases, Parasitic ; pathology ; Lysophospholipase ; metabolism ; Paragonimiasis ; pathology

In the development of diabetic nephropathy (DN), reactive oxygen specie (ROS) over much in vivo leads to oxidative stress(OS)-related renal injuries, which are characterized by the structural and functional changes in glomerular and renal tubular cells in morphology. The regulative approaches of OS involve the several signaling pathways, in which, both p38 mitogen-activated protein kinase (MAPK) signaling pathway and adenosine monophosphate-activated protein kinase (AMPK) signaling pathway play the important roles as the target of anti-oxidants. The interventional actions of Chinese herbal compound prescriptions and the extracts of single Chinese herbal medicine (CHM) on OS in the kidney in DN include regulating the balance between ROS and antioxidants, reducing the production of AGEs, inhibiting the expression of growth factors and intervening the activity of signaling pathways.
Animals ; Diabetic Nephropathies ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Kidney ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Signal Transduction ; drug effects ; Treatment Outcome

In clinic, some urinary protein makers can dynamically and noninvasively reflect the degree of renal tubular injury in patients with diabetic nephropathy (DN). These urinary biomarkers of tubular damage are broadly divided into two categories. One is newfound, including kidney injury molecule-1 (Kim-1), neutrophil getatinase-associated lipocalin (NGAL), liver-type fatty acid-binding protein (L-FABP) and cystatin C (CysC); the other one is classical, including beta2 microglobulin (beta2-MG), retinal binding protein (RBP) and N-acetyl-beta-D-glucosaminidase (NAG). It is reported that, the increases in urinary protein markers are not only closely related to the damage of tubular epithelial cells in DN patients, but also can be ameliorated by the treatment with Chinese herbal compound preparations or Chinese herbal medicine. Recently, although urinary proteomics are used in the protein separation and identification, the traditional associated detection of urinary protein markers is more practical in clinic. At present, it is possible that the associated detection of urinary biomarkers of glomerular and tubular damages may be a feasible measure to reveal the clinical significance of urinary protein markers in DN patients and the interventional effects of Chinese herbal medicine.
Biomarkers ; urine ; Diabetic Nephropathies ; complications ; drug therapy ; urine ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; methods ; Proteinuria ; complications

To analyze the characteristic of urinary protein spectrum in patients with stage III diabetic nephropathy (DN) and its compliance with traditional Chinese medicine (TCM)symptom, for the sake of providing a basis for clarifying the rules of TCM syndrome differentiation in DN. Adopting the traditional epidemiological retrospective method, thirty-eight TCM syndromes and urinary protein with medium or low molecular weight, as well as urinary enzyme, including 24 h urinary protein (Upro), urinary albumin( UAlb), urinary retinal binding protein( URBP), urinary cystatin C (UCysC), urinary N-acetyl-beta-D-glucosaminidase (UNAG), were collected from 108 patients with stage III DN, and a multiple factor regression analysis between them was conducted. As the results, the levels of Upro, UAlb, URBP, UCysC, and UNAG were increased in 108 patients with stage III DN. Qi-Yin deficiency type was the major type. The level of UAlb in patients with Qi-Yin deficiency type was significantly higher than those without Qi-Yin deficiency type (P < 0.05). The elevation of Upro with the factors as swift digestion with rapid hungering, lassitude and lack of strength, weakness of waist and knees was complied, the elevation of UA1b with the factors as dry mouth with desire to drink, the elevation of URBP with the factors as numbness of extremities, shortness of breath, the elevation of UCysC with the factors as clear urine in large amounts, and the elevation of UNAG with the factors as frequent micturition, were complied respectively. In conclusion, for 108 stage III DN patients. The increase in urinary protein spectrum including UAlb, URBP, UCysC, and UNAG is the major characteristic. Shen and Pi are the major organs related to the appearance of urinary protein; Pi-Shen deficiency is the basic pathogenesis. The level of UAlb is taken as one of the objective syndrome factors for Qi-Yin deficiency type. The levels of UNAG and UCysC are possibly the objective syndrome factors for Shen-Qi deficiency type.
Diabetic Nephropathies ; complications ; diagnosis ; urine ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Proteinuria ; complications ; urine ; Qi ; Regression Analysis ; Yin-Yang

OBJECTIVETo explore the etiopathogenisis, diagnosis and early treatment of postoperative pyogenic infection in patients with lumbar disc diseases.

METHODSFrom March 2009 to March 2012,7 patients with postoperative pyogenic infection were retrospectivly analyzed. There were 6 males and 1 female,ranging in age from 42 to 62 years old,with an average of 46.5 years old. Among 7 cases,outside the spinal canal suppurative infection occurred in 6 cases and inside the spinal canal infection in 1 case and with temporary paralysis. All the patients were treated with continuous saline lavage-drainage of low pressure impulse during operation. Unitive sensitive antibiotics were applied for 4-6 weeks after operation until CRP and ESR completely normal or the biochemistry and routine examination of the cerebrospinal fluid completely normal for the patients with intracranial pyogenic infection.

RESULTSAll the 7 cases obtained recovery and the length of stay was for 2-3 months. No remnant symptoms of nervous system were found at the leave hospital.

CONCLUSIONPostoperative pyogenic infection in patients with lumbar disc diseases is an emergency,and easily results in misdiagnosis in clinic. So the early diagnosis is very important. Early debridement is the only measure to retrieve the life of patient,continuous saline lavage-drainage of low pressure impulse may remove the remnant focus of the deep soft tissue space,and removel of the internal fixation can ensure the postoperative pyogenic infection completely control.

Adult ; Bacterial Infections ; diagnosis ; therapy ; Debridement ; Early Intervention (Education) ; Female ; Humans ; Intervertebral Disc Degeneration ; surgery ; Intervertebral Disc Displacement ; surgery ; Length of Stay ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Postoperative Complications ; diagnosis ; therapy ; Retrospective Studies

OBJECTIVEThe recombinant adenovirus vector carrying p14ARF gene was constructed for using in the interference therapy in signal transduction of laryngeal squamous cell carcinoma.

METHODSThe total cDNA fragment of p14ARF was cloned into the shuttle plasmid pAdTrack-CMV, with the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the E.Coli BJ5183 and the recombinant adenoviral plasmid was generated. The adenoviruses were packaged and amplified in the 293 cells. Then the viral titer was checked by GFP.

RESULTSThe recombinant adenovirus vector carrying p14ARF was constructed successfully. The viral titer was 2.3 x 10(9).

CONCLUSIONThe recombinant adenovirus vector could introduce p14ARF gene into the laryngeal squamous cell carcinoma line or tumor tissue effectively, which would provide experimental basis for the mechanisms and further study of the interference therapy in signal transduction of laryngeal squamous cell carcinoma.

Adenoviridae ; Cell Cycle ; Genetic Vectors ; Humans ; Plasmids ; Recombination, Genetic ; Tumor Suppressor Protein p14ARF

OBJECTIVETo explore the molecular mechanism via which the chemotherapeutic drug hydroxyurea (HU) enhances K562 cell apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL).

METHODSChronic myelogenous leukemia-derived K562 and SVT-35 cells were treated with recombinant soluble TRAIL (rsTRAIL) alone or combined with HU for a time course, and the cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-4-sulfophenyl-2H-tetrazolium-phenazine methosulphate assay. Western blot was performed to analyze the activation of apoptosis-related protein kinases and the expression of apoptosis inhibitor molecules.

RESULTSThe survival rates of SVT-35 and K562 cells treated with 1 μg/ml rsTRAIL for 24 hours were 32% and 93%, respectively. HU significantly increased the sensitivity of K562 cells to rsTRAIL cytotoxicity. Combination of rsTRAIL and HU resulted in the phosphorylation of rat sarcoma (RAS), mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK), extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase and in the significant reduction of apoptosis-inhibited molecule Fas associated death domain protein-like interleukin-1 beta-convening enzyme inhibitory protein and cellular inhibitor of apoptosis protein-1 in K562 cells.

CONCLUSIONSHU enhanced K562 cell sensitivity to rsTRAIL is mediated by Ras-MEK-ERK signaling pathway. Expression of antiapoptotic proteins cellular Fas associated death domain protein-like interleukin-1 beta-convening enzyme inhibitory protein and cellular inhibitor of apoptosis protein-1 is also down-regulated during this process. These results may through light on the therapeutic study of human chronic myelogenous leukemia.

Apoptosis ; drug effects ; physiology ; CASP8 and FADD-Like Apoptosis Regulating Protein ; metabolism ; Humans ; Hydroxyurea ; pharmacology ; Inhibitor of Apoptosis Proteins ; metabolism ; K562 Cells ; MAP Kinase Signaling System ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology

OBJECTIVETo investigate effect of interference therapy induced by epidermal growth factor receptor (EGFR)-antisense cDNA in signal transduction of Hep-2 laryngeal squamous cell carcinoma in vitro.

METHODSAdEasy Vector System was used to construct the recombinant adenovirus vector sense/antisense-pAdEasy-EGFR. The recombinant adenovirus vector introduced EGFR-sense/antisense cDNA fragment into HEK293 cell. The purified recombinant adenovirus sense/antisense-pAdEasy-EGFR transfected Hep-2 cells in vitro. The inhibition of EGFR protein expression and proliferation of Hep-2 cells, the changes of cell cycle and DNA content in Hep-2 cells were examined by MTT, Western blot analysis, flow cytometry essay, and immunocytochemistry respectively.

RESULTSThe higher titre sense and antisense mRNA expression recombinant adenovirus containing 1,032 bp EGFR-cDNA was constructed and prepared successfully. When antisense-pAdEasy-EGFR was transferred into Hep-2 cells the inhibition of cell proliferation and EGFR protein expression in Hep-2 cells were investigated effectively.

CONCLUSIONThe antisense-pAdEasy-EGFR effectively interfere the Hep-2 signal transduction pathway and induce apoptosis which results in inhibiting proliferation of laryngeal squamous cell carcinoma.

Adenoviridae ; Apoptosis ; Carcinoma, Squamous Cell ; Cell Cycle ; Cell Proliferation ; DNA, Complementary ; Genetic Vectors ; HEK293 Cells ; Head and Neck Neoplasms ; Humans ; Laryngeal Neoplasms ; Receptor, Epidermal Growth Factor ; Signal Transduction ; Transfection

OBJECTIVETo study the role of apoptosis and the expression of apoptosis-related genes Fas ligand (FasL), Fas, caspase-3 and caspase-8 in an animal model of non-alcoholic steatohepatitis (NASH).

METHODSAn experimental progressive NASH model was established by feeding male C57BL6/J mice with a high fat, methionine-choline deficient (MCD-) diet for two days, five days, ten days, three weeks and eight weeks. Control mice were fed methionine-choline supplemented (MCD+) diet. Hepatic steatosis, inflammation and fibrosis were graded by examining their H and E stained liver sections. Hepatocyte apoptosis was detected by TUNEL assay. Expressions of mRNA and protein of FasL, Fas and caspase-8 were performed by quantitative real time RT-PCR and Western blot. Caspase-3 activity assay was conducted using ApoAlert caspase-3 assay kit.

RESULTSIn MCD- mice, minimal hepatic steatosis was observed at day 5, and by day 10, mild steatosis with inflammatory infiltration was found. Severe steatohepatitis was noted at week 3, and fibrosis at week 8. TUNEL assay showed that apoptotic index in MCD- group was higher than that in MCD+ group at week 3 (15.59%+/-4.87% vs 5.17%+/-3.19%, P less than 0.05) and at week 8 (11.29%+/-3.22% vs 5.41%+/-1.54%, P less than 0.05). Compared to MCD+ group, the expression of FasL was dramatically increased on day 10 and in week 3 in MCD- mice both at the mRNA and protein levels (P less than 0.05 and P less than 0.01). Expression of Fas mRNA was up-regulated in weeks 3 and 8 (P less than 0.01), and expression of Fas in protein level was higher at week 8 (P less than 0.01) in MCD- group. Expression of caspase-8 significantly increased at the mRNA level at week 3 and week 8 (P less than 0.01 and P less than 0.05 respectively) and at the protein level at week 8 (P less than 0.05) in MCD- group. In all of the time points except for day 5, caspase-3 activities were significantly more enhanced in MCD- group than that in MCD+ group (P less than 0.05).

CONCLUSIONSIn our experimental NASH model, hepatic apoptosis was frequently detected. Increased apoptosis was probably attributable to up-regulation of apoptosis-related genes, such as FasL/Fas system, and activation of the caspase pathway. These changes may provoke hepatic apoptosis and the development of inflammation and fibrosis.

Animals ; Apoptosis ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Fas Ligand Protein ; metabolism ; Fatty Liver ; genetics ; metabolism ; pathology ; Hepatocytes ; metabolism ; Liver ; pathology ; Male ; Mice ; Mice, Inbred C57BL

Animals ; Apoptosis ; drug effects ; Cell Division ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; In Situ Nick-End Labeling ; Liver ; cytology ; drug effects ; ultrastructure