Animals ; Apoptosis ; drug effects ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; surgery ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; surgery ; Lymphatic Metastasis ; Phytotherapy ; Postoperative Period

ObjectiveToexplorethestabilityofratmodelsofsubduralhematomapreparedbysubduralinjection of different volumes of autologous blood .Methods The rats were randomly divided into sham group (36), 300μL blood group, 500 μL blood group, and 700 μL blood group (each group 60 rats).The rats of model groups received subdural injection of 300 μL, 500 μL, or 700 μL autologous blood, respectively.At the postoperative 2nd, 4th, 6th, 8th, 10th, 14th days, blood samples were taken from the abnormal aorta , and the brains were taken out for gross examination and taking photographs , six rats were used for each time .Enzyme linked immunosorbent assay ( ELISA ) was performed to determine the content of serum NSE and S100B proteins in the rats in each group.Results Compared with the sham operation group, the serum NSE in the 300μL group was significantly increased at the 2nd and 4th days (P<0.01), and then gradually reduced at the 6th and 8th days (P<0.05), indicating that the hematoma began to disappear , and at the 10th and 14th days returned to a similar level of the sham operation group (P>0.05).In the 500 μL and 700 μL blood groups, the NSE contents at 2nd, 6th, 8th, 10th and 14th days were significantly increased ( P <0.01 ), but not significantly increased at the 4th day ( P >0.05 ).The content of S100B protein in the 300 μL blood group was significantly higher at the fourth day (P<0.01), lower at the 2nd and 6th days (P<0.05), and at 8th, 10th and 14th days was similar to that in the sham operation group ( P >0.05 for all ) , indicating that the hematoma disappeared gradually, and the damages repaired .The S100B protein content of the 500 μL and 700 μL blood groups was constantly kept at a higher level ( P<0.05 ) .Conclusions Compared with the 300 μL ad 700 μL blood groups , the rat model of subdural hematoma developed by subdural injection of 500 μL autologous blood is the best , and can be used for studies of rat subdural hematoma .

Objective:To observe the efficacy, median progression-free survival (PFS) and adverse reaction induced by rh-endostatin injection (Endostar) plus platin-based chemotherapy for advanced non-small cell lung cancer (NSCLC).Methods:Fifty five histologically or cytologically confirmed advanced NSCLC patients received Endostar combined with platin-based chemotherapy for more than 2 cycles. The evaluated parameters included PFS, response rate (RR), clinical benefit rate (CBR) and adverse reaction. Results:Of the 51 patients who can be evaluated for response, 15 (29.4%) achieved partial response (PR), 27 (52.9%) had stable disease (SD), 9 (17.6%) had progressive disease(PD), no patient had complete response(CR). The overall RR was 29.4% (15/51) and CBR was 82.4% (42/51). The median PFS was 6.3 months. There were no significant differences in the short-term efficacy and PFS between the patients who had different pathological features (P=0.037), those had naive or relapsed diseases (P=0.101), or those received different chemotherapeutic regimens (P=0.232). The total white cells and platelets decreased by 72.7% and 54.5%, respectively. The frequency of grade Ⅲ or Ⅳ neutropenia and thrombocytopenia were 36.4% (20 caces) and 21.8% (12 cases), respectively. Four patients stopped the therapy for adverse reaction. One died of gastrointestinal hemorrhage; one had uncontrolled grade Ⅲ hypertension; one had superventricular arrhythmia; one had grade Ⅳ hepatic dysfunction. Conclusion:The combination of Endostar and platin-based chemotherapy increased the CBR and prolonged the PFS of the patients with advanced NSCLC. The toxicities were tolerable.

AIMTo study the binding of sibutramine hydrochloride (SH) and bovine serum albumin (BSA) in physiological condition by spectroscopic method.

METHODSThe quenching mechanism of the fluorescence of bovine serum albumin by sibutramine hydrochloride was studied with the fluorescence and the absorption spectroscopy. The binding constants K and the number of binding sites were determined at different temperatures according to Scatchard equation and the main binding force was discussed by thermodynamic equations. The effect of the drug on bovine serum albumin conformation was also studied by using synchronous fluorescence spectroscopy.

RESULTSThe quenching mechanism of sibutramine hydrochloride to bovine serum albumin was static quenching. The binding constants K at 8 degrees C, 25 degrees C, 37 degrees C were 1.21 x 10(5), 8.31 x 10(4), 6.97 x 10(4) L x mol(-1) with one binding site, respectively. The thermodynamic parameters of the reaction were deltaH = -9.70 kJ x mol(-1), deltaS = 56.41 J x mol(-1) x K(-1).

CONCLUSIONThe binding force is electrostatic interaction. Sibutramine hydrochloride can be deposited and transported by serum protein in vivo. Sibutramine hydrochloride has nearly no effect on the serum protein conformation.

Animals ; Binding Sites ; Cattle ; Cyclobutanes ; metabolism ; Protein Binding ; Serum Albumin, Bovine ; metabolism ; Spectrometry, Fluorescence ; methods ; Spectrophotometry, Ultraviolet ; methods

Objective To detect the mutations of ATP2C1 gene in patients with Hailey-Hailey dis- ease (HHD).Methods PCR and direct sequencing were performed in 17 patients and 120 healthy controls to screen the mutations in the exons of ATP2C1 gene.Results Eight mutations were identified in nine probands, including three deletion mutations (nt1464-1487 del/nt1462-1485del,1523delAT,2375delTTGT),three splice site mutations (360—2A→G,1415—2A→T,2243+2T→C) and two missence mutations (C920T and G1942T).None of the above mutations was found in the controls.Conclusion Eight specific novel mutations were identified in nine probands of HHD,which could be causative factors of the disease.

Objective To investigate the role of Sp1 in the regulation of transcription of vascular endothelial growth factor ( VEGF ) in hypoxia-induced HepG2 cells.Methods Quantitative real-time polymerase chain reaction and immunoblot assay were used to examine the effect of hypoxia on Sp1 protein and VEGF mRNA expression .Mithramycin A, the selective inhibitor of Sp1 and knock-out Sp1 gene with siRNA were used to examine the effect on VEGF mRNA expression induced by hypoxia in HepG2 cells.Results Hypoxia induced Sp1 protein and VEGF mRNA expression in HepG2 cells.Mithramycin A produced a concentration-dependent decrease of hypoxia-induced VEGF mRNA expression . After inhibition of Sp1 RNAi, VEGF mRNA expression was significantly repressed in HepG 2 cells.Conclusion Hypoxia can increase the expression of Sp1 protein and VEGF mRNA in HepG2 cells induced by hypoxia.The transcription of VEGF is regulated by Sp1 in hypoxia-induced HepG2 cells.

Obstructive nephropathy ultimately leads to end-stage renal failure. Renovascular lesions are involved in various nephropathies, and most renal diseases have an ischemic component that underlies the resulting renal fibrosis. The aim of this study was to investigate whether morphological changes occur in the renal vasculature in hydronephrosis and the possible mechanisms involved. A model of complete unilateral ureteral obstruction (CUUO) was used. Experimental animals were divided into five groups: a normal control group (N) and groups of animals at 1st week (O1), 2nd week (O2), 4th week (O4) and 8th week (O8) after CUUO. Blood pressure was measured, renal arterial trees and glomeruli were assessed quantitatively, and renovascular three-dimensional reconstruction was performed on all groups. Glomerular ultrastructural changes were examined by transmission electron microscopy. The results showed that the systolic blood pressure was significantly increased in the obstructed groups (O1, O2, O4 and O8). Three-dimensional reconstruction showed sparse arterial trees in the O8 group, and a tortuous and sometimes ruptured glomerular basement membrane was found in the O4 and O8 groups. Furthermore, epithelial media thickness and media/lumen ratio were increased, lumen diameters were decreased, and the cross-sectional area of the media was unaltered in the segmental renal artery, interlobar artery and afferent arterioles, respectively. In conclusion, renal arterial trees and glomeruli were dramatically altered following CUUO and the changes may be partially ascribed to vascular remodeling. Elucidation of the molecular mechanisms of renovascular morphological alterations will enable the development of potential therapeutic approaches for hydronephrosis.

BACKGROUND:Severe spinal angular kyphosis aggravated spinal cord injury and early degeneration, even caused incomplete paralysis or complete paralysis. Surgical treatment is the only solving approaches and method, but it is difficult, exhibits high risk, and easily affects postoperative complications. OBJECTIVE:To analyze the science and effectiveness of posterior vertebral column resection osteotomy combined with step correction in treatment of stiff angular kyphosis based on biomechanical principle. METHODS:A total of 90 cases underwent posterior vertebral column resection osteotomy combined with bilateral pedicle screw spinal cord gradual y shortening echelon tight closure and orthopedic fixation were selected, including 37 males and 52 females, at the average age of 47 years. Kyphotic angle, spinal sagittal imbalance, trunk side offset rate, operation time, intraoperative blood loss were compared and analyzed before and after treatment. RESULTS AND CONCLUSION:The kyphotic angles were 31°-138° (averagely 90.1°) preoperatively and 10°-90° (averagely 41.6°) postoperatively, with an improvement rate of 65%. The distance from C 7 plumb line to the S 1 upper edge was averagely 5.2 mm, with a correction rate of 73%. Intraoperative blood loss was 1 200-6 000 mL, averagely 2 089 mL. Operation time was 212-470 minutes, averagely 326 minutes. The patients were fol owed up for 20 to 35 months after the surgery. Osteotomy segments had achieved bone fusion in al patients, and no complications of spinal cord injury or orthopedic angle loss appeared. These data verified that in the accordance with cellbiomechanics and spinal biomechanical principles, bilateral pedicle screw spinal cord gradual y shortening echelon tight closure and orthopedic fixation protected utmost spinal cord cells against injury in the correction of thoracolumbar angular kyphosis. There is sufficient basis for cellphysiology and it accorded biomechanical and physiological characteristics. During the surgery, we should pay attention to protection and release of nerve root and avoid postoperative corresponding nerve root irritation. Ful fusion ensures kyphosis correction and avoids spine lateral offset, is an effective safeguard for the recovery of spinal function and postoperative orthopedic effect.

BACKGROUND:The treatment difficulties of thoracolumbar angular kyphosis surgery are:low correction rate, hard to rebuild sagittal plane, easily induce neurological complications, postoperative loss of balance, high incidence of pseudarthrosis and postoperative loss of correction degree.
OBJECTIVE:To explore the safety and efficacy of modified posterior vertebral column resection osteotomy and bilateral pedicle screw combined with echelon tight closure spinal cord technique and implant fixation for severe spinal angular kyphosis.
METHODS:A total of 87 severe spinal angular kyphosis patients, 36 males and 51 females, who were treated in the Department of Orthopedics, the 306 Hospital of Chinese PLA from January 2006 to December 2013, were enrol ed in this study. They underwent posterior vertebral column resection, bilateral pedicle screw combined with echelon tight closure spinal cord, and implant fixation. Kyphosis, spinal sagittal imbalance, offset rate towards trunk side, operation time and intraoperative blood loss were observed before and after treatment.
RESULTS AND CONCLUSION:The preoperative average kyphosis was 90.1° (31°-138°). The postoperative average kyphosis was 27.9° (15°-57°). The improvement rate was 76%. The improvement rate of trunk sagittal offset was 76%. Intraoperative blood loss was 800-3 000 mL, and average blood loss was 2 300 mL. The operation time was 5-7 hours, averagely 5.9 hours. Before treatment, two patients affected neurologic symptoms in double lower extremity, and their Frankel classification was grade C and became grade E after treatment. Al patients were fol owed up for 9-57 months. Bony fusion was achieved in al patients. No complications of spinal cord injury appeared, and no orthopedic angle missing occurred. These results indicate that during posterior vertebral column resection for treating severe angular stiffness of the thoracic kyphosis, blood vessels could be maintained greatly. Blood vessel injury-induced ischemic changes in spinal cord and ischemic reperfusion injury could be avoided. To reduce hemorrhage and to keep effective blood volume in patients with low body mass are effective for early recovery after treatment. Bilateral pedicle screw combined with echelon tight closure spinal cord technique greatly protected spinal cord cells against injury. We should pay attention to the protection and loose of nerve root to avoid postoperative nerve root irritation. Sufficient bone fusion ensures kyphosis correction, avoids spine lateral offset, and plays a key role in spinal function and postoperative orthopedic effect.

Adiponectin is an adipocyte-derived secretory protein. It was found to be associated with insulin resistance, inflammation and arteriosclerosis. To further study the biological function and expression of adiponection in vivo, adipoenctin gene knock-out and LacZ gene knock-in mouse model was constructed. Gene targeting strategy was designed to replace part of exon 2 and exon 3 of adiponectin gene with full length LacZ gene in frame with remaining upstream ATG and signal peptide sequence of exon 2. The targeting vector (Adipo-LacZ-XpPNT) was constructed and verified by restriction enzyme digestion and sequencing. CJ7 ES cells were transfected with targeting vector linearized by NotⅠ digestion, selected in the medium containing both G418 and ganciclovoir. Resistant clones were screened by PCR and further confirmed by Southern blot for correct homologous recombinants. Chimera mice were obtained by routing microinjection of homologous recombined ES cells into blastocysts. After mating, mice heterozygous and further homozygous for adiponectin knockout and LacZ gene knock-in were established. Expression of both endogenous adiponectin and exogenous LacZ gene in mouse tissues and sera were detected by RT-PCR, Northern-blot, Western blot and ELISA. The results show that adiponectin was disrupted at both mRNA and protein levels. LacZ gene is expressed exclusively in adipose tissue of mutant mice. Its expression profile is identical to endogenous adiponection. Unexpectedly, LacZ activity could not be detected in both adipose tissue and serum although LacZ protein can be detected in adipose tissue but not in serum of mutant mice. In conclusion, mice homozygous for adiponectin knockout and LacZ gene knock-in have been successfully constructed. Mutant mice display LacZ expression profile identical to endogenous adiponectin albeit neither LacZ activity nor protein can be detected in serum of mutant mice.