Objective To investigate wheter epigallocatechin gallate (EGCG) could prevent the information of abrant crypt loci (ACF) induced by isoquinoline (IQ)and its possible mechanisms. Methods Sixty male BALB/cA nude immunological deficit mice were divided into five groups. Except control group, the other four groups were received IQ to induce ACF. The rats in low, medium and high dose groups were received 5, 10 and 20 mg/kg of EGCG,respectively. The mice were sacrificed six weeks later. Hematoxylin-eosin (HE) staining and 0.2% methylene blue staining were used to observe the routine histology and ACF, respectively. Immunohistochemistry (IHC) was used to detect Nrf2 protein level and RT-PCR was used to detect Nrf2 and UGT1A10 mRNA levels in colon tissue. Results The body weights of model group decreased significantly compared to high-dose group (21.70±0.13 vs. 24.37±0.07, P<0.01). Compared to model group, the degree of atypical hyperplasia and even canceration of colon mucus and the number of total ACF and total AC in high-dose group were decreased significantly (18.00±7.51 vs. 64.20±45.18, P<0.05;63.90±18.58 vs. 168. 80±35.34, P<0.01). The protein level of Nrf2 increased (0.3114±0.0037 vs. 0.1660±0.0021, P<0.01). The mRNA levels of Nrf2 and UGT1A10 in high-dose group was increased (both P value<0.01). Conclusions EGCG has protective effect on IQ induced preneoplastic lesions through reducing the number of ACF. This effect may be caused partly through the signal pathway Nrf2-UGT1A10.

BACKGROUND: As the damage caused by protein glycation is one of the mechanisms of diabetes, it is helpful to treat diabetes related diseases with the understanding of the inhibition of berberine on protein glycation and the protection to the brain damage caused by protein glycation.OBJECTIVE: To observe the effects of berberine on glycated brain damages induced by D-galactose in model rats.DESIGN: Randomly grouping paralleled control study.SETTING: Department of Ophthalmology, Xiamen Traditional Chinese Medicine Hospital.MATERIALS: The experiment was conducted in the Department of Pharmacology, Pharmacy College of Jinan University from June to October 2005. Ninety SD rats (6 weeks old) were selected and divided into 6 groups: control group, model group, hydrochloride aminoguanidine group and high (300 mg/kg), middle (150 mg/kg) and low (75 mg/kg) doses berberine groups with 15 rats in each group. The glycated models were established by intraperitoneal injection of D-galactose. The main drugs:berberine was from Guangzhou Wanji Drugs Limited Company; D-galactose was from Shanghai Yuanju Bioscience Technology Limited Company.METHODS: The rats in the control group were intraperitoneally injected the normal saline for 8 weeks; rats in other groups were injected 5%D-galactose (150 mg/kg) for 8 weeks. From the 3rd week, the hydrochloride aminoguanidine group was infused hydrochloride aminoganidine (150 mg/kg); the three doses berberine groups were given corresponding doses berberine; the control group and model group were given distilled water for 6 weeks with the volume of 10 mL/kg. At the end of the 8th week, the erythrocyte aldose reductase activity was determined by coomassie brilliant blue method; the level of plasma glycohemoglobin was measured by thio-barbituric acid colorimetry and the fructosamine in serum was measured by nitroblue tetrazolium colorimetry. The quantity of advanced glycation end products (AGEs) in serum, and AGEs, malondialdehyde (MDA), and activity of superoxide edismutase (SOD) in brain tissue and calcium ion in neurons were also dertermined. Moreover, the changes of mitochondria in brain hippocampus cells were observed under electronic microscope.MAIN OUTCOME MEASURES: ① The AGEs, plasma glycohemoglobin, serum fructosamine and aldose reductase activity. ②AGEs in brain tissues. ③Calcium level in brain. ④MDA content and SOD activity in brain tissues. ⑤Changes of mitochondria in hippocampus neurons.RESULTS: All 90 animals were involved in the result analysis. ①Aldose reductase activity and glycated product content in serum: After the rats were treated with D-galactose for 8 weeks, the aldose reductase activity in red blood cells and the content of fructosamine in serum, glycohemoglobin,AGEs in the model group were significantly higher than those in the normal control group (P ＜ 0.01); After treated by high and middle doses berberine for 6 weeks, the activity of aldose reductase and content of fructosamine in serum (absorbancevalue of hemoglobin every 10 g), glycohemoglobin, and AGEs were obviously lower than those in the control group [(1.07±0.39), (1.22±0.47), (1.76±0.30) nkat/g, t=5.052, 5.484, P ＜ 0.01;(0.740±0.142), (0.862±0.131), (0.958±0.083) mmol/L, t=7.829, P ＜ 0.01,t=2.404, P ＜ 0.05; 58.434±12.135, 64.614±13.418, 83.747±7.990,t=4.922, 6.748, P ＜ 0.01; (3.104±0.814), (2.937±0.514), (4.156±0.860) U/mg,t=4.104, 3.440, P ＜ 0.05]; the aldose reductase activity of the low dose berberine group was lower than the model group (P ＜ 0.05), which had no obvious effect on glycated products. ②AGEs in brain tissues: The contents in the hydrochloride aminoganidine group, high and middle doses berberine groups were lower than the model group [(10.52±1.22), (10.95±1.75),(11.95±2.27), (14.26±3.51) U/mg, t=-3.892, -3.263, P ＜ 0.01, t=-2.139,P ＜ 0.05], and the low dose berberine had little effect (P ＞ 0.05). ③Calcium level in neurons: The levels in the hydrochloride aminoganidine group,and high dose berberine groups were lower than the model group.[(271.52±32.71), (293.84±31.58), (337.15±58.49) nmol/L, t=-3.421, P＜ 0.01, t=-2.275, P ＜ 0.05], the low dose berberine group had no obvious effect (P ＞ 0.05). ④MDA content and SOD activity in brain tissues: MDA contents in the hydrochloride aminoganidine group, high and middle doses berberine groups were lower than the model group, and the SOD activity was markedly higher than the model group [(2.09±0.16), (2.12±0.22),(2.41±0.12), (2.54±0.21) μmol/g, t=6.601, 5.348, P ＜ 0.01, t=2.082, P＜ 0.05; (8.79±1.09), (8.80±1.52), (7.90±1.48), (6.48±1.34) mkat/g, t=4.571,4.254, P ＜ 0.01, t=2.226, P ＜ 0.05]. ⑤Mitochondria structure in brain hippocampus cells: Under the electronic microscope, mitochondria in brain hippocampus cells of the model group appeared obvious swelling with broken crests and disorganized structure, even obvious big vacuoles were observed. In the hydrochloride aminoganidine, and high and middle doses berberine groups, no obvious swelling was observed with vacuoles only in a few mitochondria. Nevertheless, obvious swelling appeared in mitochondria of low dose berberine group with broken crest and disorganized structure,and vacuoles were observed.CONCLUSION: D-galactose-induced damage in mitochondria may be related to AGEs formation in brain tissue, maladjustment of calcium ions in neurons and oxidative stress in rat models. Berberine can inhibit glycation induced by D-galactose and protect rat brain tissues from glycated damage.

AIM:To investigate the effects of aminoguanidine intervention on lens cell damage induced by D-galactose in rat eyes and its mechanism of action.METHODS:D-galactose(400 mg/kg) was injected into rats intraperitoneally for 14 weeks to induce the animal model of glycosylation and lens cell damage.Aminoguanidine(75 mg/kg,150 mg/kg) were administered for 12 weeks by intragastric administration beginning at 3rd week.All animals were killed and blood samples were taken to measure the activity of aldose reductase,the level of fructosamine,the amounts of glycohaemoglobin and advanced glycation end-products.The lenses of eyes were taken to detect the activities of AR,GR,SOD and SDH.The amounts of AGEs,GSH,MDA or outleakage of LDH were measured,respectively.The ultrastructure and apoptosis of lens epithelial cells were examined by transmission electron microscope and flow cytometry,respectively.RESULTS:Animals were treated with D-galactose for 14 weeks,the serum level of fructosamine,the amounts of glycohaemoglobin and AGEs,and activity of AR were significantly increased.The amount of AGEs and activity of AR in lens were increased,the activity of antioxidase was decreased and oxidative product was increased.The apoptosis,the damages of mitochondria and cell nucleus in lens cells were observed.After treated with aminoguanidine for 12 weeks,the activity of AR and the level of fructosamine in serum,and the amounts of glycohaemoglobin and AGEs were significantly decreased(P

Objective To investigate the preventive effects of epigallocatechin gallate (EGCG) on growth and metastases of orthotropic colonic cancer. Methods Forty BALB/C male nude mice were prepared for model of colonic cancer and then divided into control group and low-, medium- and high-dose of EGCG groups with 10 each. Except control group, the mice in other three groups were treated with 5, 10 and 20 mg·kg~(-1)·d~(-1) of EGCG. The effect of EGCG on growth and metastases of colonic cancer was observed. The histopathologic changes of liver and lung were observed with HE, and protein expression of NF-E2-related factor 2 (Nrf2) in cancerous tissue was detected by immunohistochemistry. RT-PCR was used to examine mRNA levels of Nrf2, UDP-glucuronosyhrans-ferase (UGT)1A, UGT1A8 and UGT1A10. Results In comparison with control group [(564±130) mg], the average weight of the tumor in low-, medium- and high-dose groups was (152±63) mg, (76±42) mg and (18±10)mg, respectively, with tumor inhibitory rate of 73.0%, 86.5% and 96.8%, respectively (all P value<0.05). There was a positive correlation between tumor inhibitory effect and dosage of EGCG (P<0.05). The protein expression of Nrf2 and the mRNA levels of Nrf2, UGT1A, UGTIA8 and UGTIA10 in three EGCG treated groups were increased significantly compared with control group (P<0.05), and there was a phenomenon of nuclear transcription of Nrf2. Conclusions EGCG can prevent local growth and metastases of orthotopic colonic cancer in a dose-dependent manner in nude mice. The inhibitory effect may be caused by inducing the expressions of Nrf2, UGT1A, UGT1A8 and UGT1A10 genes.

This study was aimed to establish a method for simultaneous determination of seven anions and organic acids in Huo-Xue Tong-Luo (HXTL) injection by HPCE. With tartaric acid as the internal standard, separation was performed on an uncoated fused silica capillary (50 μm × 64. 5 cm, 56 cm of effective length). The 14 mmol·L-1 potassium acid phthalate and 0.1 mmol·L-1 hexadecyl trimethyl ammonium chloride were selected for the running buffer solution (pH 5.6). The separation voltage was -16 kV. The detection wavelength was set at 210 nm. The column temperature was maintained at 25 ℃ . The sample was injected at 50 mbar×4 s. The results showed that
calibration curves of chloride ion, sulfuric acid root ion, formate ions, malic acid, succinic acid, iodate ion and acetic acid ions showed good linear relationship 41.4-248.2 μg·mL-1 (r = 0.999 3), 12.5-74.8 μg·mL-1 (r = 0.999 8), 18.2-109.1 μg·mL-1 (r = 0.999 8), 20.3-121.6 μg·mL-1 (r = 0.999 5), 17.2-103.1 μg·mL-1 (r=0.999 1), 17.6-105.6μg·mL-1 (r=0.999 6), 51.6-309.6μg·mL-1 (r=0.999 7), respectively. The average recoveries were 102.6%, 97.3%, 102.2%, 99.0%, 99.2%, 97.8%, and 103.4%, respectively. The RSD were 1.7%, 2.0%, 1.6%, 2.6%, 2.1%, 2.9%, and 1.0%, respectively (n = 6). It was concluded that the method was accurate and reproducible. It was suitable for the determination of anions and organic acids in HXTL injection.

Objective To identify the RNA interference action of recombined pSUPER-NRF2 vectors for the expression of NRF2 gene in colon cancer cells.Methods Two sequences targeting at the ORF of NRF2 were cloned into RNA polymerase III based expression vector pSUPER.These recombinants were transfected into Caco-2 cells.Fluorescence microscopy and flow cytometry were performed after transfection with pEGFP-N1 plasmids to observe the lipfectin transfection efficiency.The stable cells were selected in medium(48 h) after co-transfected pEGFP-N1 with G418. The expression of NRF2 was assayed by RT-PCR and Western blot.Results The construction of the recombinant expression vector pSUPER-NRF2-A1、B1 and its control vector pSUPER-NRF2-A2、B2 were successfully confirmed by the results of enzyme digestion,electrophoresis and sequencing. The transfection efficiency was 45.6%,74.3%,53.0% and 46.5% respectively in 24,48,72 and(96 h).We compared the ability of these vectors to inhibit NRF2 in a transient and stable expression experiment.Importantly,pSUPER-NRF2-B1 was able to knockdown NRF2 expression.pSUPER-NRF2-A1 only had a moderate activity,whereas pSUPER-NRF2-A2、(B2 were) inactive in this assay.Conclusion The constructed pSUPER-NRF2-A1、B1 showed an interfering effecton the expression of NRF2 and product the stable cells with low NRF2 expression.Therefore,the pSUPER vector constitutes a new and powerful system to analyze NRF2 gene function in colon cancer.

ObjectiveTo evaluate the efficacy and safety of tirofiban in treatment of ST segment elevation acute myocardial infarction(STEAMI) no-reflow and acute thrombosis after emergency percutaneous coronary intervention(PCI). MethodsForty patients which were made definite diagnosis of STEAMI were intra-coronary artary injection fortirofiban after emergency PCI stenting occured no-reflow and acute thrombosis. First,the dose of 0.4μg·kg-1·min-1 was given from intra--coronary artary injection of tirofiban within three minutes, after 30min the dose were given 0.1μg·kg-1·min-1 for 48 hours. ResultsThe no re-flow and acute thrombosis was completely disappeared within five minutes,at the time,side effect with in one week was not observed. ConclusionsTirofiban treatment by direct injection in coronary arteries combined with emergency PCI, can increase the repeffusion rate of infarction related vessel in AMI patients,and improve TIMI reflow. This reperfusion method was effective and safe.

CBS(Case Based Study)teaching method is given in a case requiring students to answer a series of questions surrounding the case.In the teaching process,students of small groups are required to find their own solutions.Compared with tradition method,this mothed can improve students'academic performance(P

BACKGROUND:Procolagen type 1 N-terminal propeptide (P1NP) and β-colagen special sequence(β-CrossLaps) are two bone metabolic markers that are closely related to osteoporosis. Combined detection of bone metabolic markers and bone mineral density is of clinical significance for the diagnosis of osteoporosis. Bone metabolic markers are ideal indicators to predict fractures, which can compensate for the lack of bone density test. OBJECTIVE:To introduce the application of bone metabolic markers in the monitoring of drug efficacy on the treatment of osteoporosis as wel as in the prediction of fracture risks in recent 20 years and to explore the clinical values of P1NP and β-CrossLaps to assess the therapeutic efficacy on osteoporosis and risks for osteoporotic fractures. METHODS:A computer-based search of CNKI and SCI databases were performed for relevant articles published from 2000 to 2014 using the keywords of “serum bone metabolic markers; osteoporosis; bone mineral density” in Chinese and English, respectively. Finaly, 44 articles meeting the inclusive criteria were reviewed. RESULTS AND CONCLUSION:This paper analyzes the source and detection mechanisms of P1NPand β-CrossLaps and then compares their advantages in the therapeutic effect assessment of osteoporosis. Serum bone metabolic markers cannot only reflect the dynamic changes of bone metabolism, but also have earlier changes than the bone mineral density. Both P1NPand β-CrossLaps are very important for assessing the early diagnosis of osteoporosis as wel as anti-osteoporosis drug efficacy.

BACKGROUND: The nutritional support, as well as the water and electrolyte balance during the perioperative period in the renal transplantation recipients at diuresis stage are important to the functional restoration of transplanted kidneys.OBJECTIVE: To explore the method and opportunity of the nutritional support and the handling of the water and electrolyte balance in perioperative period of renal transplantation.DESIGN, TIME AND SETTING: A retrospective clinical analysis was performed in the Department of Urology, Xijing Hospital from June 2003 to June 2007.PARTICIPANTS: Ninety-six patients of chronic renal failure underwent allograft renal transplantation. They comprised 59 males and 37 females, aged 17-67 years, with a mean of 35.7 years.METHODS: The perioperative physiological features of the renal transplantation recipients were summarized retrospectively. The recipients' condition during the perioperative period was divided into two stages at the opening point of allograft blood current. The vital signs of the patients maintained at a stable level before operation. All patients received blood transfusion since the operation began, and were supplemented with albumin before opening the vessels. Urinary production exceeding 100 mL per hour indicated the beginning of fluid replacement, which was a simplified transfusion for the patients at diuresis stage following renal transplantation.MAIN OUTCOME MEASURES: Blood inosine, urea nitrogen, electrolyte, blood sugar and urine of the patients were detected at one day postoperatively.RESULTS: During 12-16 hours postoperatively, the urinary production was 260-1 200 mL, average 520 mL per hour. Blood routine test showed 8 cases developed mild hyponatremia, accounting for 8.3%, 3 cases occurred high potassium and healed after renal functional recovery, 1 case presented low potassium and healed with supplement therapy. There were no abnormal changes of blood chlorine. The blood glucose among 21 cases (21.9%) was higher than the normal level, and recovered following hormone maneuver. The electrolytes and blood glucose were detected to be normal in other patients, without any case with low calcium or magnesium. The urine specific gravity arranged during 1.010-1.015.CONCLUSION: The colloid such as erythrocytes, blood plasma and albumin should be mainly infused before the opening of allograft blood current. And the water and electrolytes is recommended to administrate promptly and regularly during the diuresis stage. The healing of the stoma benefits from the adequate nutritional support. The metabolic acidosis still should be prevented when the urinary production returns normal.