OBJECTIVE: To observe the expression pattern of caspase-3 and HCLS1-associated protein X-1 (HAX-1) at different time after cerebral contusion in rat, and explore the new method for estimating the injury interval. METHODS: The cerebral contusion model was established using adult SD male rats. Then the rats were randomly allocated into 8 groups: 2 h, 6 h, 12 h, 1 d, 3 d, and 7 d after cerebral contusion, sham-operation and normal control. Expression of caspase-3 and HAX-1 protein after cerebral contusion in rat was detected by Western blotting. Laser scanning confocal microscope was used to observe the number of HAX-1 positive cells and TUNEL-stained cells after cerebral contusion. RESULTS: The expression of caspase-3 increased parallelly with the time after cerebral contusion and reached the peak value on 3 d. The expression of caspase-3 decreased gradually and still maintained a high level expression on 7 d (P < 0.05). The expression of HAX-1 positive cell went up after injury, and reached the peak value at 6 h (P < 0.05), then turned down gradually after 12 h and went out of detection after 3 d. The number of TUNEL-stained cells increased obviously at 2 h and reached the peak value on 3 d. The number of TUNEL-stained apoptotic cells decreased gradually and still maintained a high level expression on 7 d (P < 0.05). CONCLUSION The expression of caspase-3 and HAX-1 after cerebral contusion has time sequential regularity, which may provide new evidence for forensic diagnosis of cerebral contusion interval.
Animals ; Blotting, Western ; Brain Injuries/pathology* ; Carrier Proteins/metabolism* ; Caspase 3/metabolism* ; Cerebellum/pathology* ; In Situ Nick-End Labeling ; Intracellular Signaling Peptides and Proteins ; Male ; Rats ; Rats, Sprague-Dawley

The study was using the orthogonal test and making the extraction rates of icariin, ferulic acid, epimedin A, epimedin B, epimedin C, baohuoside I and ligustilide determinated by HPLC multiwavelength switch, gradient elution and multi-index comprehensive weighted scoring method (weight coefficient was 0.47: 0.16: 0.07: 0.07: 0.08: 0.06: 0.09) as evaluation index, combine with SPSS 16.0 software to optimizing the best extraction. It was Yinpian soak 1 h, 12 times more than the volumn of 50% ethanol solution, by heating reflux extraction for 60 min. The compliance test indicates that the optimized compatibility extraction technology is stable and practical, and it has provided an experimental basis for compound preparation technology research of Epimedium brevicornu and Ligusticum chuanxiong.
Chemical Fractionation ; methods ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Epimedium ; chemistry ; Ligusticum ; chemistry

Genomic DNA was extracted from the blood samples of 3 patients from 1 pedigree with congenital nephrogenie diabetes insipidus (NDI) and their 12 family members.The whole coding region of the arginine vasopressin receptor 2 (AVPR2) gene was amplified by PCR and then directly sequenced,A mutation of AVPR2 gene [g1236T→C (L292P)]was found in 3 patients.The patients' mothers were found to have both mutant and normal alleles.

To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.
Administration, Oral ; Animals ; Arylamine N-Acetyltransferase ; genetics ; metabolism ; Caffeine ; metabolism ; urine ; Cytochrome P-450 CYP1A2 ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Female ; Male ; Medicine, Tibetan Traditional ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Theophylline ; urine ; Uracil ; analogs & derivatives ; urine ; Uric Acid ; analogs & derivatives ; urine ; Xanthines ; urine

OBJECTIVE: To investigate the expression of caspase-3 and iNOS in different intervals and to provide evidence for estimation of injury intervals after brain contusion in human. METHODS: Thirty cases died of serious brain injury were included into the injury groups and 5 cases died of non-brain injury were served as control group. To analyze the changes of caspase-3 and iNOS expression in brain samples at different intervals (2h, 4-8h, 10-14h, 1-2d, 3-5d, 8-11d) by immunohistochemistry and auto-image analysis system. RESULTS: The level of caspase-3 expression started to increase in 2 hours after brain contusion compared to the control group (P<0.05). The level of caspase-3 expression continued to increase in 1-2 days and maintained high level in 3-5 days compared to the control group (P<0.05), then decreased gradually. There was no statistically significant difference between the expression level of iNOS in 2 hours with the control group (P>0.05). But the expression level of iNOS began to increase in 4-8 hours after brain contusion and reached its maximum in 1-2 days, then decreased. Weak expression of iNOS still could be detected in 8-11 days. CONCLUSION The expression of caspase-3 and iNOS can be used as effective evidence for human brain contusion interval.
Adult ; Astrocytes/metabolism* ; Brain/pathology* ; Brain Injuries/pathology* ; Caspase 3/metabolism* ; Female ; Forensic Pathology ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neurons/metabolism* ; Nitric Oxide Synthase Type II/metabolism* ; RNA, Messenger/metabolism* ; Staining and Labeling ; Time Factors ; Young Adult

BACKGROUNDIIIb-T(4) non-small cell lung cancer (NSCLC) is commonly considered a contraindication to surgery, although chemo-radiotherapy also achieves a poor survival rate. We reviewed our experience with T(4) NSCLC patients who underwent surgery to explore the indications and prognostic factors of surgical treatment of lung cancer invading the left atrium and great vessels.

METHODSWe investigated a cohort of 105 patients, 79 men and 26 women, who underwent surgery from May 1996 to July 2008. Their pathological staging was T(4)N(0)-(2)M(0). The median age was 59 years, ranging from 36 to 75 years. Patients were grouped based on invading sites: tumors invading the left atrium (LA group), tumors invading the superior vena cava (SVC group), and tumors invading the intrapericardial pulmonary artery (PA group). Patients were further characterized based upon the type of operation, complete resection and incomplete resection groups, and on the lymph node pathological status, N(0), N(1) and N(2) groups. We calculated the overall five-year survival rate.

RESULTSAll patients received resection of primary lesions, with partial resection of the left atrium in the LA group (n = 25), angioplasty of superior vena cava in the SVC group (n = 23) and intrapericardial ligation of the pulmonary artery in the PA group (n = 57). Complete resection was possible in 77 patients (73.3%). The overall survival rate of the 105 patients was 41.0% at 5 years; 36.0% for the LA group, 34.8% for the SVC group and 45.6% for the PA group. Pathological N status significantly influenced the overall 5-year survival rate; 61.5% for N(0), 51.1% for the N(1) and 11.8% for the N(2) groups (N(2) group versus N(0) group, P < 0.0001, N(2) versus N(1) group, P < 0.0001). Surgical resection also influenced survival; 49.4% for the complete resection group and 17.9% for the incomplete resection group (P < 0.0001). Cox regression analysis demonstrated that pathological N status was a significant independent predictor of prognosis.

CONCLUSIONSPathological N status is a significant independent predictor for survival of patients with IIIb-T(4) lung cancer invading the left atrium and great vessels. The completeness of resection has a significant influence on the overall 5-year survival rate. Surgery for T(4) lung cancer may be effective in patients without mediastinal lymph node involvement.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; complications ; surgery ; Female ; Heart Atria ; pathology ; Humans ; Lung Neoplasms ; complications ; surgery ; Male ; Middle Aged ; Neoplasm Invasiveness ; pathology ; Survival Rate ; Treatment Outcome

OBJECTIVETo investigate the effects of Rehmannia glutinosa oligosaccharides (ROS) on the proliferation of HepG2 and insulin resistance.

METHODThe HepG2 cells were divided into control group, rosiglitazone (3.4 mg x L(-1)) treated group and ROS (0.1-30 mg x L(-1)) treated group. The proliferation of HepG2 was detected by MTT method. Insulin resistant HepG2 cells model was induced by high concentration of insulin, then the effects of ROS on glucose consumption in insulin resistant HepG2 cells were investigated.

RESULTIn the middle glucose culture medium, the absorbance at 570 nm of HepG2 was increased by high concentration of ROS, and decreased by low concentration of ROS by using MTT method, a concentration-dependent manner. ROS increased glucose consumption in HepG2 cells, and showed a better effect at the dose of 10 mg x L(-1). ROS promoted the glucose consumption in insulin resistance of HepG2 cells, improved the sensitivity of insulin resistance of HepG2 cells to insulin.

CONCLUSIONHigh concentration of ROS can promote the proliferation of HepG2, and however low concentration of ROS inhibits the proliferation of HepG2. ROS can significantly improve insulin resistance of HepG2 cells induced by high insulin.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; Glucose ; metabolism ; pharmacology ; Humans ; Hypoglycemic Agents ; isolation & purification ; pharmacology ; Insulin ; pharmacology ; Insulin Resistance ; Liver Neoplasms ; metabolism ; pathology ; Oligosaccharides ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rehmannia ; chemistry

OBJECTIVETo investigate clinical effects of coracohumeral ligament reconstruction with autologous double-strand of long palmaris longus tendon and artificial ligament for the treatment of acromioclavicular joint dislocation.

METHODSFrom April 2006 to June 2009, 31 patients with acromioclavicular joint dislocation were treated with coracohumeral ligament reconstruction using autologous double-strand palmaris longus tendon and artificial ligament. There were 18 males and 13 females, ranging in age from 18 to 60 years, with an average of 35 years. Twenty-six patients were acute trauma and other 5 patients were chronic trauma. Preoperative symptoms included different degrees of pain, restricted movement, and instability of acromioclaviecular joint. The X-ray showed acromioclavicular joint dislocation.

RESULTSThe patients had good incision union without vascular and nerve injuries. All the patients were followed up, and the average duration was 23 months. The JOA scores decreased from preoperative (38.8 +/- 1.5) to (73.2 +/- 1.1) at 1 month after operation,and (93.5 +/- 0.8)at the last follow-up. Twenty-eight patients got an excellent result, 2 good and 1 fair.

CONCLUSIONThe reconstruction of coracohumeral ligament using autologous double-strand palmaris longus tendon and artificial ligament is an effective method for the treatment of acromioclavicular joint dislocation.

Acromioclavicular Joint ; injuries ; physiopathology ; surgery ; Adolescent ; Adult ; Artificial Organs ; Clavicle ; Female ; Follow-Up Studies ; Humans ; Joint Dislocations ; physiopathology ; surgery ; Ligaments, Articular ; physiopathology ; surgery ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Scapula ; Tendons ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the changes of large conductance Ca(2+)-activated K(+) channel (BK channel) on coronary smooth muscle cells from diabetic rats.

METHODSStreptozotocin-induced rat diabetic animal model was used. Coronary smooth muscle cells were isolated by enzyme digestion. BK currents in control and diabetic groups were recorded by patch clamp technique in whole cell configuration, and BK channel protein expression was detected by Western blot. Calcium concentration was measured by fluorescence assay.

RESULTSCompared with control group, BK current densities in diabetic group were significantly decreased when test potentials > 100 mV (P < 0.05). BK current densities were (275 ± 40) pA/pF in control group (n = 8) and (70 ± 10) pA/pF in diabetic group (n = 6) at 150 mV test potentials. α-subunit protein expression was similar between the groups (P > 0.05), however, β1-subunit protein expression was significantly reduced in diabetic group than in control group (P < 0.05). Calcium concentrations were significantly increased in diabetic group control group (151 ± 18) nmol/L (n = 6) than in control group (92 ± 7) nmol/L (n = 5, P < 0.05).

CONCLUSIONObserved β1-subunit downregulation, BK current density decrease and cytosolic calcium concentration increase in smooth muscle cells of diabetic coronary arteries may be associated with coronary dysfunction in diabetic rats.

Animals ; Calcium ; metabolism ; Coronary Vessels ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; Large-Conductance Calcium-Activated Potassium Channels ; metabolism ; Male ; Muscle, Smooth, Vascular ; metabolism ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the protective effects of estrogen on the mitochondria in human umbilical vascular endothelial cells (HUVECs).

METHODSHUVECs were exposed to H2O2 at 250 micromol/L for 4 h with or without pretreatment with 17-estradiol (E2) and ICI182780. Complex IV activity of the cells was measured with chromometry, and 2, 7-dichlorofluorescein diacetate (DCFH-DA) was used to determine intracellular reactive oxygen species (ROS). Intracellular adenosine triphosphate (ATP) level was quantified with a luciferin- and luciferase-based assay.

RESULTSCompared to the blank control group, H2O2 caused a decrease in complex IV activity, intracellular ATP level, and the cell viability, but elevated intracellular ROS. E2 pretreatment of cells significantly attenuated these effects of H2O2 exposure. ICI182780 administered prior to E2 pretreatment antagonized the protective effects of E2 against H2O2 exposure.

CONCLUSIONE2 offers mitochondrial protective effects on HUVECs, which is mediated by the estrogen receptors.

Cells, Cultured ; Cytoprotection ; drug effects ; Electron Transport Complex IV ; metabolism ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Estrogens ; pharmacology ; Female ; Humans ; Hydrogen Peroxide ; pharmacology ; Mitochondria ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Pregnancy ; Reactive Oxygen Species ; metabolism ; Umbilical Veins ; cytology