Objective To investigate the antitumor immunity induced by tumor derived mixed heat shock protein/peptide(mHSPs),interleukin-12(IL-12)and Cyclophosphamide(Cy).MethodsPurified mixed HSP was prepared from tumor by S180 protein extraction and purification,SDS-PAGE,Western blot and animal experiment were applied for mixed HSPs analysis.ResultsThe proliferation of cytotoxic T lymphocyte(CTL)cocultured in the mHSPs+Cy+IL-12 group was significantly remarkable and the content of CD8+ CTLs was significantly more in comparison with the other groups(P<0.01).To the tumor bearing mice,mHSPs+Cy+IL-12 group showed partial therapeutic efficacy,the averaged survival period was over 60 d,and 90% of the mice in this group got long period tumor free survival(>90d),obvious difference(P<0.05)from the other groups.ConclusionTumor derived mixed HSPs can induce powerful antitumor immune efficacy and show favorable therapeutic efficacy.

OBJECTIVE To map a comprehensive metabolic pathway of herbacetin in rats, specifically, to elucidate the biotransformation of herbacetin in vivo and to simultaneously monitor the pharmacokinetic process of both parent drug and its major metabolites. METHODS liquid chromatography/ion trap mass spectrometry (LC/MSn) and ultra-liquid chromatography coupled with mass spectrometry (UPLC/MS) were combined in the current study for qualitative and quantitative determinations of herbacetin and its metabolites in bile, urine and feces after both oral and intravenous administration of herbacetin to rats. Enzyme kinetic studies on the intestinal and hepatic metabolism of herbacetin were further conducted to elucidate metabolic profiles of herbacetin in rat tissues and organs. Additionally, plasma concentration profiles of herbacetin and its metabolites in rats were obtained to characterize the overall pharmacokinetic behavior of herbacetin. RESULTS It was found that herbacetin was excreted primarily from rat urine in the form of glucuronide-conjugations. Subsequent in vitro enzyme kinetic studies and in vivo pharmacokinetic investigations suggested an extensive hepatic metabolism of herbacetin and the high exposure of herbacetin- glucuronides in systemic circulation. The clearance, half- life and bioavailability of herbacetin in rats were determined as (16.4±1.92)mL·kg-1·min-1, (11.9±2.7)min, and 1.32%, respectively. On basis of these findings, a comprehensive metabolic pathway of herbacetin in rats was composed. In addition, a physiology based pharmacokinetic (PBPK) model was successfully developed with the aid of the GastroPlus to simulate the pharmacokinetic process of herbacetin in rats. Application of the PBPK modeling can provide a useful starting point to understand and extrapolate pharmacokinetic parameters among different species, populations, and disease states. CONCLUSION After oral administration, herbacetin was subjected to colonic degradation and extensive first pass metabolism, with glucuronidation as its dominating in vivo metabolic pathway.

OBJECTIVE To investigate the effect of hypoxia on the pharmacokinetic process of salidrosidein rats and to explore its underlying mechanisms. METHODS The Caco-2 cell monolayerwas exposed to 1% oxygen (O2) concentration for 24 h to build the hypoxiccell model. The transportation mode of salidroside was investigated with the aid of this hypoxia model by detecting the apparent permeability coefficient(Papp). Healthy Sprague Dawley (SD) rats were exposed to 9% O2 for 72 h for the construction of hypoxic rat model. Liver sample was subsequently collected from the hypoxic rats with an aim to identify enzymes responsible for salidroside metabolism. The expression levels of sali?droside-transporting and salidroside-metabolizing enzymes, including Sodium-dependent glucose cotrans?porters (SGLT1), β-glucosidase (GBA3)and sulfotransferase (SULT2A1), were thereafter detected by RT-PCR and Western blot. The metabolic activity of GBA3 and SULT2A1 was monitored by rat liver microsome incubation.In addition, the renal function of rats under hypoxia was assessed by detecting concentrations of blood urea nitrogen and creatinine. RESULTS The AUC and t1/2 values of salidroside in hypoxic rats were more than doubled, while the in vivo clearance was significantly reduced. Mechanistic study demonstrated that the PappA- B/PappB- A eualsto 10.3, indicating the potential active transport of salidrosile. The expression of SGLT1 and GBA3 was significantly decreased, which indicated a reduced metabolism of salidroside under hypoxia. Moreover, rat under hypoxia was found to suffer from renal dysfunction, with an abnormal value of blood urea nitrogen. CONCLUSION Due to the reduced metabolism and the abnormal renal function under hypoxia, the systemic exposure of salidroside in rats was signifi?cantly enhanced.

Objective To explore the feasibility and effectiveness of interventional transcatheter destruction of the aortic valve to establish an animal model with acute aortic valve regurgitation. Methods Eight healthy goats were used for this study. A limited sternotomy approach was used to access the apex of the heart. Puncturing of the apex of the heart was performed to establish a wire track, then, under fluoroscopic guidance a 10 F sheath was inserted along this track of hard wire until to the ascending aorta above the aortic valve. The internal sheath was removed. Via the 10 F sheath a 10 mm occluder of ventricular septal defect (VSD) was introduced into the ascending aorta above the aortic valve. The sheath was pulled back to the left ventricle, while the occluder remained in the ascending aorta above the aortic valve. Then the occluder was quickly pulled back into the left ventricle in order to make some certain damage to the aortic valve. And an acute aortic valve regurgitation model was thus established. Angiography of ascending aorta above the aortic Among the 8 animals, two died of acute left ventricular failure on the spot due to excessive regurgitation blood flow after the operation. Macroscopically, damage of the aortic valve was seen. In the six survivors, angiography of ascending aorta above the aortic valve and Doppler echocardiography showed that moderate degree of regurgitation was detected in 5 and small amount of regurgitation in one. Two experimental goats with moderate degree of regurgitation died of heart failure separately at seven days and fifteen days after the operation. The remaining four experimental goats survived for more than three months. Follow- up checkups with echocardiography suggested the presence of mild- moderate degree of regurgitation. Conclusion Acute aortic valve regurgitation model in experimental goats can be established through transapical transcatheter damage of aortic valve by quickly pulling back a VSD occluder which has been placed in the ascending aorta above the aortic valve. This method is clinically feasible, technically simple and repeatable, the result is reliable, and the degree of regurgitation is controllable.

GATA transcription factor family members have been found to involve in the growth and differentiation of mammary gland. Among them GATA-3 is regarded as the most critical regulator involving the tumorigenesis of breast cancer (BC). Recently, trichorhinophalangeal syndrome-1 gene (TRPS-1), a new GATA family member, has been identified to be highly prevalent in breast cancer. Compared with ER-negative breast cancer, the expression of TRPS-1 is higher in ER-positive breast cancer and was significantly correlates with estrogen receptor, progesterone receptor, and GATA-3, indicating it may serve as a ductal epithelial cell-specific regulator in the differentiation of breast ductal epithelial cells. Studies have shown that miR221/222 is able to downregulate the expression of an epithelial cell marker E-cadherin by targeting TRPS-1, resulting in mammary epithelial cells transition to mesenchymal cell (EMT). In addition, it has been well accepted that, and the Science and Technology Bureau of Jiaxing (2012AY1071-2)TRPS-1 plays a role in the differentiation of several other cell types including kidney nephric mesenchymal cells, columnar chondrocytes, and osteoclasts, indicating that TRPS-1 involves in mesenchymal-to-epithelial cell transition (MET). In this article, we summarize the roles of GATA transcription factor TRPS-1 in ductal epithelial cells and the roles of its gene and protein expressions in predicting the prognosis of breast cancer.
Breast Neoplasms ; genetics ; pathology ; DNA-Binding Proteins ; genetics ; Epithelial-Mesenchymal Transition ; Female ; GATA3 Transcription Factor ; genetics ; Humans ; Transcription Factors ; genetics

The study of sarcosaphagous insects is a subspecialty in forensic medicine based on the knowledge of entomology. It could help to determine the time of death, especially the postmortem interval in decomposed cases. This paper explores its history, species and erosion process of sarcosaphagous insects. It reviews the species identifying methods with molecular biology and entomological morphology. Details of its application in estimating postmortem interval in recent years and study of sarcosaphagous insects in the field of forensic medicine are summarized.
Animals ; Cadaver ; Death ; Diptera/physiology* ; Entomology/methods* ; Forensic Medicine/methods* ; Humans ; Larva/growth & development* ; Postmortem Changes ; Time Factors

Objective To study the effect of low-frequency suprathreshold repetitive transcranial magnetic stimulation (rTMS) of the unaffected hemisphere on recovery of motor function in patients with acute stroke. Methods A total of 26 patients with middle cerebral artery territory infarction were randomly assigned to unaffected hemisphere stimulation group and control group (not receiving any stimulation,n=13).The patients in the stimulation group were treated with rTMS 3 to 5 d after the onset of symptoms with the frequency of 1 Hz and 70％ of the intensity (about 2.1T actual output) and the 1200 pulses per day for 10 consecutive d.The motor evoked potential (MEP) latency,central motor conduction time (CMCT),scores of National Institutes of Health Stroke Scale (NIHSS) and modified Barthel index (MBI) of the affected brain region were recorded on the 1 st of experiment (before the treatment),10 and 40 d after treatment. Results The scores of clinical futction scale and neuroelectrophysiologic parameters before treatment had no statistical significance between the 2 groups (P＞0.05).The scores of clinical function scale after the treatment in the 2 groups were obviously higher than those before treatment (P＜0.05). And the improvement of motor function in the unaffected hemisphere stimulation group was statistically obvious as compared with that in the control group (P＜0.05):the score of NIHSS and the MBI in the stimulation group were obviously higher than those in the control group (P＜0.05).The neuroelectricity physiological indexs in the 2 groups after treatment gained improvement in comparision to those before treatment:the MEP latency on the 40th d of treatment and CMCT on the 10th and 40th d of treatment in the unaffected hemisphere stimulation group was significantly different as compared with those before treatment (P＜0.05); the CMCT on the 10th and 40th d of treatment in the unaffected hemisphere stimulation group was shorter as compared with that in the control group. Conclusion The frequency of 1 Hz and 70％ of the intensity (about 2.1T actual output) in rTMS of the unaffected hemisphere can shorten CMCT and improve the motor function in patients with acute stroke.

OBJECTIVEFungal keratitis (FK) is a vision-threatening infection, whose treatment requires more effective and safer anti-fungal agent exploitation urgently. With this aim, we focused on the effect of an extracellular polysaccharide on fungal adhesion to human corneal epithelial cells.

METHODSWe performed the cytotoxicity assays of the extracellular polysaccharide EPS-II from an antarctic bacterium Pseudoaltermonas and evaluated its inhibitory effect on Candida albicans cells' adherence to human corneal epithelial cells (HCECs).

RESULTSEPS-II, which displayed minor cytotoxicity but also promoted proliferation of HCECs, could inhibit the adherence of yeast cells to HCECs in a dose-dependent manner. EPS-II could also suppress the subsequent PI3K/AKT signaling pathway, and thereby decrease the expression of early inflammatory cytokines.

CONCLUSIONSExtracellular polysaccharide EPS-II was suggested as a new natural agent for attenuating FK.

Blotting, Western ; Candida ; drug effects ; physiology ; Cell Adhesion ; drug effects ; Cornea ; microbiology ; Humans ; Phosphorylation ; Polysaccharides, Bacterial ; pharmacology ; Proto-Oncogene Proteins c-akt ; metabolism ; Pseudoalteromonas ; metabolism

OBJECTIVETo investigate the effect of vascular bundle implantation in autogenous bone graft on angiogenesis.

METHODSThirty-six New Zealand white rabbits were evaluated in this study. A portion of bilateral radial bones of a rabbit were removed as free bone grafts, whose periostea were peeled off. In test group, the external maxillary artery bundle was passed through the marrow cavity of the bone. In control group, there was no vascular bundle implantation. Each bone was placed in masseter muscle separately. The rabbits were sacrificed and the specimens were procured at 3 days, 1, 2, 3, 4 and 6 weeks after surgery for histological observation, Chinese ink perfusion and CD34 immunohistochemistry. Microvessel density (MVD) was assessed in order to evaluate angiogenesis of autogenous bone grafts.

RESULTSThe bone grafts were found revascularization in 3 days after surgery in the test group, whereas at 2 weeks in the control group. In 3 days, 1 week, 2 weeks, 3 weeks and 4 weeks after surgery, the MVD of test group was significantly higher than that of control group. In 4 weeks after surgery, angiogenesis of test group reached to peak.

CONCLUSIONVascular bundle implantation improved angiogenesis in non-vascularized autogenous bone graft in this study.

Animals ; Bone Transplantation ; Bone and Bones ; Prostheses and Implants ; Rabbits

OBJECTIVETo research the efficacy and feasibility for unstable fracture of thoracolumbar with AF spine internal fixation device.

METHODSThirty-two patients with unstable fractures of T11-L3 were treated with AF spine internal fixation device and autograft between vertebral lamina vertebral body transverse process from January 2002 to June 2006. There were 21 female and 11 male, aging from 58 to 72 years with a mean of 62 years. All these patients were examined with x-ray and CT preoperative and postoperative respectively. They were followed-up thirteen months averagely, observing the stability of spinal column, bone grafting fusion, the height of vertebra and recovery of anterior bone fragment herniation.

RESULTSAll these AF spine internal fixation devices treated for the unstable fractures of thoracolumbar had not removed because of internal fixation failure or pain. Fracture healing and grafting fusion appeared after operation three months averagely. X-rays revealed post-protrusion angle were recovered from 22 degrees to 8.5 degrees, the heights of anterior were recovered from 50% to 86%, the angle of posterior were recovered from 94% to 98%. The postoperative CT scan showed that six cases with herniation to canal gained a completely recoveries.

CONCLUSIONAF spine internal fixation device used in early stage for unstable fracture of thoracolumbar is a simple and effective method. It has advantages such as providing early substantial fixation, maintaining a well three column stability. Bone grafting is a key factor in this operative technique.

Aged ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; surgery