Objective:Guanxinshutong on cerebral ischemia reperfusion ( I/R) in rats learning and memory ability and brain Bcl-2,Bax protein expression to explore the protection mechanisms of Guanxinshutong on cerebral I /R injury in brain tissue.Methods:Healthy Wistar rats for the study were randomly divided into sham operation group ,I/R model group,high dose Guanxinshutong (1 g/kg) and low dose Guanxinshutong (0.5 g/kg) group,n=15.cerebral I/R model was estabhished by using the left commom cartid artery ligation method,modeling each group were fed daily before saline ,saline,Guanxinshutong (1 g/kg) and Guanxinshutong (0.5 g/kg), continuous 7 days;3 d after water maze training while continuing administration ,continuous 7 d,the eight day ,Morris water maze test;after the test,the rats were sacrificed blood and brain tissue ,serum by ELISA of Bcl-2 and Bax protein levels change;using HE staining of rat brain pathology;using immunohistochemistry assay in rat brain organization of Bcl-2 and Bax protein levels changes.Results:Compared with the sham group ,Guanxinshutong high-dose group test latency slightly longer timers after platform and platform quadrant dwell decreased slightly ,serum and brain tissue slightly elevated levels of Bax protein ,Bcl-2 protein levels decreased slightly but not significantly different ( P>0.05 ) ,low-dose group Guanxinshutong through latency test in rats ,after the number of platforms and platform quadrant dwell time decreased , serum and brain tissue elevated levels of Bax protein , Bcl-2 protein content decreased significantly different(P<0.01),cerebral I/R model group was significantly prolonged latency test ,after the number of platforms and platform quadrant dwell time significantly reduced serum and brain tissue was significantly increased Bax protein ,Bcl-2 protein content decreased with significant differences ( P<0.01 ).Conclusion: Guanxinshutong can significantly improve spatial memory , and by reducing Bax protein content and increased Bcl-2 protein,inhibition of cerebral I/R injury of apoptosis,and thus the brain tissue I/R has a protective effect.

OBJECTIVE:To establish a method for the contents determination of tetrahydropalmatine and rosmarinic acid in Xiaoheling capsule. METHODS:HPLC was performed on the column of Phenomenex C18 with mobile phase A of 0.1% phosphoric acid(adjusted pH to 6.0 with triethylamine)-methanol(45:55,V/V) and (adjusted pH to 6.0 with triethylamine) B of 0.1% phos-phoric acid-methanol(55:45,V/V)(gradient elution)at flow rate of 0.8 ml/min,detection wavelength was 282 nm(0-25 min)and 330 nm(25.01-60 min)and volume size was 10 μl. RESULTS:The linear range was 1.502-75.10 μg/ml for tetrahydropalmatine and 6.03-301.6 μg/ml for rosmarinic acid(r=0.999 8);RSDs of precision,reproducibility and stability tests were ≤1.36%;recoveries were 97.11-99.87%(RSD=0.95%,n=9)and 98.47-101.99%(RSD=1.08%,n=9),respectively. CONCLUSIONS:The method is simple,accurate and reliable,and can be used for contents determination of tetrahydropalmatine and rosmarinic acid in Xiaoheling capsule.

Objective: To investigate the effects of simulated altitude acute hypoxia on appetite and orexin mRNA expression in rats. Methods: Rats were exposed to hypobaric chambersimulating different high altitude (3000m,4000m,5000m,6000m) for different times (8h/d;1d,3d and 7d respectively), and the changes of appetite, weight gain, and the orexin mRNA expression in hypothalamus of rats were examined. Results: With the increasement of hypoxia time and simulated altitude, the total food intake of rats and the orexin A and B mRNA expression in hypothalamus decreased gradually as compared with control. After 7 days of hypoxia, the weight gain of rats was significantly different(P

This study aimed at exploring the effects of Griffipavixanthone,a natural small molecule extracted from Garcinia esculenta,on the autophagy and the mechanism behind it.Cervical cancer HeLa cells were selected as the model in vitro.Confocal microscopy,flow cytometry and immunoblotting were performed to know the impacts of Griffipavixanthone on signaling pathway over autophagy flux and calcium level;while nude mouse model was established to explore its anti-cancer action in vivo.It was found that Griffipavixanthone significantly induced GFP-LC3 puncta formation and the accumulation of LC3-Ⅱ/LC3-Ⅰ,and increased the protein expression of SQSTM1 in HeLa cells in a time and concentration dependent manner.MTT assay results showed that Griffipavixanthone inhibited the growth of HeLa cells in a dosage dependent manner,and eliminated the tolerance of tumor cells to nutrient starvation,which fastened the death of cells.Griffipavixanthone also down-regulated the concentration of calcium in the cytoplasm.Furthermore,the in vivo study indicated that intratumoral injection of Griffipavixanthone presented the anti-cancer effect to a certain extent.In conclusion,it was suggested that Griffipavixanthone inhibit autophagy with the potential to be developed into a clinical anti-cancer agent.Griffipavixanthone sensitized HeLa cells to nutrient stress-induced cell death through inhibiting the autophagy flux and the level of calcium in the cytoplasm.

Objective:To detect the expressions of survivin, galectin-3 and thyroid peroxidase in papillary thyroid microcarcinoma ( PTMC) and cancer adjacent normal tissue to explore the clinical significant.The correlations between the expressions of survivin,Gal-3 and TPO with BRAFV600E gene mutation in PTMC were analyzed.Methods: The expressions of survivin,Gal-3 and TPO were measured by immunohistochemical ( IHC ) method in 56 cases of PTMC tissue and adjacent normal tissue; BRAFV600E mutation was detected by PCR amplification and subsequently sequencing.Chi square test was used to analyse the relation in the expression rates of survivin,Gal-3 and TPO and BRAFV600E gene mutation.Results:The survivin and Gal-3 were strongly expressed but TPO was negatively expressed.The survivin and Gal-3 were negative in adjacent normal tissues but TPO was expressed.There were sig-nificant differences in the expression rates of survivin, Gal-3 and TPO between PTMC tissue and adjacent normal tissue (all P<0.001).The BRAFV600E mutation rate (32.1%) and the expression rates of survivin,Gal-3 and TPO in PTMC tissue were found to be positively related to lymph node metastases (P=0.009,P=0.025,P=0.007,P=0.008),and negatively related to gender and age (all P>0.05).There were no correlation was found between the expressions of survivin,Gal-3,TPO and the BRAFV600E gene mutation in PTMC(all P>0.05).Conclusion: The strong expressions of survivin and Gal-3,the mild expression of TPO and BRAF mutation may be important in the development of PTMC,and the detection of BRAFV600E gene mutation and the expressions of survivin, Gal-3 and TPO could be used to the judgment of pathogenetic condition and prognostic outcomes.

Bone Neoplasms ; pathology ; Cerebellum ; pathology ; Humans ; Neurocytoma ; pathology ; Petrous Bone ; pathology

In this study, a novel tetrapeptide hydrophilic interaction chromatography (HILIC) material was synthesized, and corresponding enrichment method for glycopeptides was developed.The tetrapeptide modified silica gel materials (denoted as Poly-DAPD) were synthesized by atom-transfer radical-polymerization (ATRP) and characterized by N2 adsorption desorption and thermometer, thermal gravimetric analyzer (TGA) and X-ray photoelectron spectrometer (XPS).The characterization results indicated that tetrapeptide had been successfully synthesized on silica gel.Poly-DAPD materials showed high enrichment selectivity toward fetuin glycopeptides under solid-phase extraction (SPE) mode.Comparing with commercialized ZIC-HILIC in glycopeptides enrichment of fetuin digest which mixed with 5 mole ratio of albumin bovine (BSA), the as-prepared materials showed higher selectivity in both aspects of the identified number of glycopeptides and anti-interference property.The SPE results demonstrated that the tetrapeptide-based HILIC materials could be a potential tool in large-scale glycosylation analysis.

This study was intended to establish a method of purification of HPV16 L1 protein expressed in a prokaryotic system and to obtain the purified protein. The prokaryotic expression vector pGEX-4T-1-HPV16 L1 was constructed and transformed into E. coli BL21 cell, and induced by 1 mM IPTG to express HPV16L1 protein. The inclusion bodies were isolated and solubilized with 8 M urea. After the urea was removed by gradual dialysis, the denatured L1 protein were renatured and then were purified by affinity chromatography. The results showed that HPV16L1 protein formed inclusion bodies in bacterial expression system, suggesting that this assay can be used to purify HPV16L1 protein and hence provide a basis for studying the applications of HPV16 L1 protein.
Capsid Proteins ; biosynthesis ; Escherichia coli ; metabolism ; Genetic Vectors ; Human papillomavirus 16 ; Oncogene Proteins, Viral ; biosynthesis

Objective To investigate the expression and significance of γH2AX in cervical squamous carcinoma.Methods Firstly,DNA were extracted from 74 cervical squamous carcinoma samples and PCR were tested for HPV infection.Secondly,formalin-fixed paraffin-embedded tissue sections (4 μm)were stained with H&E method to detect cervical lesions grading.Thirdly,HPV16 DNA were examined by in situ hybridization(ISH) and γH2AX,p16 were examined by immunohistochemical (IHC) staining.Then,30 cases typical tissue sections in which including the normal cervical tissue,cervical intraepithelial neoplasia and cervical carcinoma in situ were selected for comparing the HPV DNA loading,and the γH2AX and,pl6 expression.Finally,the feasibility of γH2AX serving as a biomarks in HPV infection-related cervical carcinogenesis were analyzed.Results In this study,HPV infection ratio is 98.65％,and HPV16 is the most common type with 74.32％ infection.In situ hybridization showed no HPV16 DNA exist in normal cervical tissues and CINI.In CIN Ⅱ HPV DNA exist mainly as episomal DNA.With the increasing of cervical lesions grade,HPV DNA was integrated into chromosome steadily.The expression of γH2AX and pl6 were positively associated with grading of cervical lesions.HPV DNA and γH2AX protein co-exist primarily in the prickle cell layer and the granular cell layer.The HPV DNA and p16 protein exist in different cell layer.Conclusion γH2AX may be employed as a biomarker for HPV positive cervical carcinogenesis.

Measuring the length of the proportional units (PU) of different truncal parts in males and females and analyzing the relationship between these measurements showed that there was nosignificant gender difference in the length of one cun respectively for the perpendicular proportional units (PPU) of prothorax, epigastrium and flank but there were significant differences in the PPU of the hypogastrium, lateral thorax and hypochondrium and the horizontal proportional unit (HPU) of the back; within the same gender, there was no significant difference in the length ofone cun between the PPU of the prothorax and that of epigastrium and no significant difference betweenthe PPU of the flank and the HPU of the intermammillary region in males but there was a significant difference between PU of other parts.