Carcinoma, Hepatocellular ; genetics ; pathology ; Gene Expression Profiling ; Genomics ; Humans ; Liver Neoplasms ; genetics ; pathology ; Neoplasm Metastasis ; genetics ; Neoplasm Recurrence, Local ; genetics

To establish the online two-dimensional liquid chromatography by using double gradient liquid chromatography system and UV detector, in order to simultaneously determine the content of paeoniflorin, paenol, amygdaloside and cinnamic acid. A pump of the two-dimensional liquid chromatography was adopted as the one-dimensional separation pump. C18 (3.0 mm x 150 mm, 3 microm) was used as the analytical column, with acetonitrile as the organic phase and 0.08% phosphoric acid + 0.08% triethylamine as the aqueous phase for gradient elution at the flow rate of 0.5 mL x min(-1). Another pump of the two-dimensional liquid chromatography was adopted as the two-dimensional separation pump. PAII C18 was used as the analytical column, with acetonitrile as the organic phase and 20 mmol, pH 3.0 monopotassium phosphate as the aqueous phase for gradient elution at the flow rate of 0.8 mL x min(-1). The detection wavelengths were set at 218, 230, 275 nm by using wavelength time-switching program. The linearity range of paeoniflorin, amygdaloside, paeonol and cinnamic acid were 5.55-222 (r = 0.999 7), 6.6-264 (r = 0.999 8), 3.3-132 (r = 0.999 5) and 0.315-12.6 mg x L(-1) (r = 0.999 7), respectively. The average recoveries of the four components were between 96.12% and 103.9%. The experiment proved that this method was so rapid and accurate in determination results that it could be used for evaluating drug quality.
Capsules ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Online Systems ; Time Factors

OBJECTIVETo explore the scavenging action of tenuigenin (TEN) on intracerebral amyloid β protein (Aβ) aggregation and the abnormal phosphorylated tau protein and its mechanism in Alzheimer's disease (AD) rats' brain.

METHODSAβ1-40 was injected into the right CA1 region hippocampus to establish the AD model. Successfully modeled rats were divided into the model group, the low, middle, high TEN group. Rats were administered with TEN (18.5, 37.0, 74.0 mg/kg) by gastrogavage. Besides, a sham-operation group was set up. Expression levels of Aβ1-40 and Tau p-Ser262 were detected by immunohistochemistry. Expression levels of ubiquitin (Ub) and Ub-protein ligase E3 were measured by Western blotting.The content of 26S proteasome was detected by ELISA.

RESULTSImmunohistochemical results showed that the number of Aβ and Tau p-Ser262 positively reacted neurons significantly increased in model group, when compared with the sham-operation group (P < 0.01). Results of Western blot showed expression levels of ubiquitinated protein were up-regulated and those of Ub-protein ligase E3 were down-regulated in the model group (P < 0.01). ELISA results showed that the content of 26S proteasome significantly decreased in AD rats' brain (P < 0.01). Compared with the model group, expression levels of Aβ1-40, Tau p-Ser262, and Ub significantly decreased; expression levels of Ub-protein ligase E3 apparently increased; the content of 26S proteasome significantly increased in each TEN treatment group (P < 0.05, P < 0.01). Best effect was shown in 37.0 mg/kg and 74.0 mg/kg TEN groups.

CONCLUSIONSUb proteasome pathway (UPP) participated in the occurrence of AD. TEN could obviously reduce intracere- bral Aβ1-40 accumulation and abnormal tau phosphorylation.

Alzheimer Disease ; metabolism ; Amyloid beta-Peptides ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Hippocampus ; metabolism ; Neurons ; metabolism ; Phosphorylation ; Proteasome Endopeptidase Complex ; metabolism ; Rats ; Ubiquitin-Protein Ligases ; metabolism ; Ubiquitins

Objective To investigate the clinical significance of procalcitonin(PCT) in early diagnosis of neonatal infections.Methods Rapid hemi-quantitative immunoassay was used to measure PCT levels in 196 hospitalized neonates, who were divided into sepsis group,(local-)infection group, virus-infection group and non-infection group.Results If plasma PCT≥0.5 ?g/L was taken as positive,the positive rates in sepsis group,local-infection,virus-infection,non-infection group were 87.87%,41.66 %,12.0%,11.11%,respectively.The positive rate of sepsis group was significantly higher than that of the other 3 groups(all P

Objective To explore the relationship bet-ween air pollution and acute onset of cerebral hemorrhage in Hangzhou.Methods Time-stratified case-crossover study was used to analyze the effect of aerosol optical depth(AOD),PM10,SO2 and NO2 on the acute onset of cerebral hemorrhage.Conditional logistic regression was used to estimate the odds ratios(Ors) and their 95% confidence intervals(Cis) in relation to an increase of one unit of AOD and 10 μg/m3 of air pollutants.Results After adjusted temperature and relative humidity,the Ors of acute onset of cerebral hemorrhage by a unit increase in AOD at a 2 day-lag were 1.727(95%CI:1.103-2.703)in first half year and 2.412 (95%CI:1.230-4.733) at a 2 day-lag in spring.For a 10 μg/m3 increase in SO2,the Ors were 1.119(95% CI:1.019-1.229),1.230(95%CI:1.092-1.386),1.254(95%CI:1.076-1.460) in the whole year(2 day-lag),in first half year(2 day-lag) and in spring(2 day-lag),respectively.NO2 exposure in first half year(2 day-lag) was associated with cerebral hemorrhage,with OR as 0.841(95% CI:0.734-0.964).However,there were no statistical significances for AOD,SO2,NO2 in the rest time-periods(P>0.05).Additionally,no association was found between PM10 and the acute onset of cerebral hemorrhage in any time-periods(P>0.05).Conclusion Our data showed that there was association between air pollution and the acute onset of cerebral hemorrhage,especially in spring and in the first half of the year.

Objective:To summarize our experience on perioperative anesthesia management of patients with crush syndrome caused by earthquake under field condition.Methods:The clinical anesthesia data of 10 patients with crush syndrome caused by China Wenchuan earthquake,who were treated under field condition,were retrospectively analyzed,and our experience on perioperative anesthesia management was summarized.Results:After volume therapy,urine alkalization,and stabilization of internal environment,the 10 patients underwent dehridement,decompression by fasciotomy or amputation under epidural anesthesia or brachial plexus block plus psychological therapy and moderate tranquillizer.During the operation,the patients showed smooth respiratory and cardiovascular characteristics without pain.Conclusion:Under field condition,regional anesthesia can provide adequate analgesic effect during and after the surgery.Early fluid resuscitation and life support are vital to the safety and life of patients.

OBJECTIVETo explore relationship between volume of bone cement injection and concurrent of fracture after thoracolumbar osteoporotic vertebral fracture treated by percutaneous kyphoplasty (PKP).

METHODSFrom January 2006 to December 2008,68 patients with thoracolumbar osteoporotic vertebral fracture treated by PKP were retrospectively analyzed. Among them, 30 patients with less than 3 ml bone cement injection (mean 2.5 ml, low group), including 11 males and 19 females, with an average age of (85.0 +/- 8.5) years (ranging for 60 to 91); 38 cases with over 4 ml bone cement injection (mean 4.5 ml, large group), including 15 males and 23 females,with an average age of (86.0 +/- 9.2) years (ranging for 60 to 93). Factors of concurrent vertebral fractures were observed during follow-up.

RESULTSAll patients were followed up from 3.4 to 5.1 years with an average of 3.8 years. Thirteen patients (43.3%) co-occurred fracture in low group,among which strengthened concurrent vertebral fracture occurred in 1 case,upper and lower section adjacent vertebral fracture in 8 cases,distal segment of vertebral fracture in 4 cases; while 18 patients (47.3%) co-occurred fracture in large group,among which strengthened concurrent vertebral fracture occurred in 2 cases, upper and lower section adjacent vertebral fracture in 10 cases,distal segment of vertebral fracture in 6 cases. No significant difference between two groups (P > 0.05).

CONCLUSIONBone cement injection is not main influence factors for treating concurrent of fracture after thoracolumbar osteoporotic vertebral fracture by PKP. Concurrent fracture mainly relates with progress of osteoporosis, the volume of injection volume may appropriately over the volume of balloon.

Aged ; Aged, 80 and over ; Bone Cements ; Female ; Follow-Up Studies ; Humans ; Injections ; Kyphoplasty ; adverse effects ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Osteoporotic Fractures ; etiology ; Postoperative Complications ; etiology ; Retrospective Studies ; Thoracic Vertebrae ; injuries ; surgery

Objective To assess the efficacy of two antibiotic prophylactic regimens in a prospective randomized trial in 1 year for patients undergoing insertion of catheters,and to provide the evidence for uniform consensus existing on the timing,route,and choice of antibiotic.Methods During a period of 12 months,78 patients,who consecutively entered the peritoneal dialysis programme,[45 women and 33 men,mean age (48.2?15.7)years] were included.The prophylactic regimens were a single dose of ceftriaxone (1.0 g) given intravenously 30 minutes before surgery (Group A) and given cefazolin (0.25 g/L) i.p.in the each dialysis bag for 3 days postoperatively (Group B).All operations were performed in one room.The wound was observed every day,and body temperature,Count of white blood corpuscle and type,dialysate were examined every day. Results In Group A and B,none of the patients showed peritonitis or wound infection during the post-operative period (within 10 days).One of 39 patients(2.5%) in the group A,and 2 of 39 patients (5.1%) in the group B had exit site infection (P＞0.05).Conclusions There is no significant difference in the incidence of peritonitis and wound infection between two groups. Prophylactic preoperative single-dose antibiotics intravenously do as well as antibiotics given intraperitoneally for peritoneal dialysis catheter insertion,but is much more convenient.

Cystatin, which widely distributed in both tissues and body fluids of animal and plant, was a superfamily of cysteine proteinase inhibitors. It could form activity-inhibitor complexes with cysteine proteinases to inhibit the hydrolytic activity of proteinases. Cystatin played important roles not only in the inhibition of the proteolytic degradation of fish muscle, but also in biological defense systems against invaders. To explore the functions of fish cystatin and the potential values in fish disease prevention and cure, as well as seafood processing, the recombinant yeast strains which could express Chinese sturgeon cystatin were constructed. First, the cystatin cDNA of Chinese sturgeon, which had been PCR modified, was subcloned into yeast integrated vector pPICZaA. After extracted and purified, the recombinant plasmids were linearized by Sac I. The yeast Pichia pastoris GS115 strain was transformed by use of the Lithium Chloride transformation method, and the recombinant cystatin yeast strains got. After 0.5% methanol induction, SDS-PAGE analysis of the culture supernatant indicated that the yield of recombinant cystatin was about 215mg x L(-1) with the percentage about 73.6%. The recombinant cystatin was purified through Q-Sepharose anion-exchange chromatography, and the purity reached about 94.2%. The inhibitory activity of recombinant cystatin was measured by inhibiting the proteinase activity of papain. The results showed that about 1 microg recombinant cystatin could inhibit the activity of 15 microg papain. Heat stability assay results showed that there was a decrease in inhibitory activity of cystatin with the increasing of temperature. When solution of recombinant cystatin was kept at 70 degrees C for 5min, the inhibitory activity reduced fast. While the recombinant cystatin was heated to 90 degrees C for 5min, the inhibitory activity of recombinant cystatin was undetected. The inhibitory activity for recombinant Chinese sturgeon cystatin was higher than that of CPI (cysteine proteinase inhibitor) from seeds of corn, that about 1 microg purified CIP could inhibited the activity of 0.278 microg papain. But the heat stability of recombinant cystatin is lower than that of the corn CPI. The expression level and the activity of recombinant cystatin from yeast Pichia pastoris were higher than those from E. coli. Moreover, recombinant cystatin from Pichia pastoris was easier to separate and purify. This paper reported that recombinant fish cystatin was produced in a highly efficient expression system based on the methylotrophic yeast, further work will focus on the function of recombinant Chinese sturgeon cystatin to resist fish disease and explore the value of cystatin as a food additive to inhibit cysteine proteinases during surimi processing.
Animals ; Cystatins ; genetics ; metabolism ; pharmacology ; Cysteine Proteinase Inhibitors ; genetics ; metabolism ; pharmacology ; Enzyme Activation ; drug effects ; Fish Proteins ; genetics ; metabolism ; Pichia ; genetics ; metabolism ; Polymerase Chain Reaction ; Protein Stability ; Temperature

BACKGROUNDCathespin-B (cath-B) is an important proteolytic enzyme involved in the disease course of invasion in many types of cancer. Cath-B expression in subcutaneous heteroplastic pancreatic carcinoma in nude mice has not been studied. We investigated the role of cath-B in a model of heteroplastic pancreatic carcinoma in BALB/c nude mice.

METHODSThirty-two six-week-old female BALB/c nude mice were equally divided into four groups. PANC-1 cells were inoculated subcutaneously in the left axillary region. Besides volume, weight of subcutaneous tumor, and change in body weight, cath-B expression in each group was measured by immunohistochemical staining, PCR and Western blotting. Its relationship to microvessel density (MVD), CD44v6, and placenta growth factor (PLGF) was also examined. CA-074Me, a specific inhibitor of cath-B, was injected intraperitoneally (i.p.) at different stages of tumor growth in group B and C. Gemcitabine (GEM), was also injected (i.p.) in group D to compare anti-tumor efficacy with CA-074Me.

RESULTSExpression of cath-B at different levels was related to tumor growth, MVD, and PLGF expression. In group A (control group), cath-B expression was enhanced more than that seen in other groups. CA-074Me clearly inhibited cath-B expression and tumor growth in group B. There was no difference between group C and D with respect to anti-tumor effect.

CONCLUSIONSCath-B correlates with the growth and angiogenesis of tumors, but not with the adhesion induced by CD44v6. CA-074Me clearly inhibited cath-B expression and demonstrated an anti-neoplastic and anti-angiogenesis effect.

Animals ; Antineoplastic Agents ; therapeutic use ; Blotting, Western ; Body Weight ; Cathepsin B ; antagonists & inhibitors ; genetics ; metabolism ; physiology ; Cell Line, Tumor ; Dipeptides ; therapeutic use ; Female ; Humans ; In Vitro Techniques ; Mice ; Mice, Nude ; Pancreatic Neoplasms ; drug therapy ; metabolism ; Placenta Growth Factor ; Pregnancy Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Heterologous