Objective To evaluate the effect of methylene blue pertreatment on acute kidney injury induced by sepsis in rats.Methods Ninety male Sprague-Dawley rats,aged 1.5-2.5 months,weighing 200-250 g,were randomly assigned into 3 groups (n =30 each) using a random number table:sham operation group (S group),cecal ligation and puncture (CLP) group and methylene blue group (MB group).The rats were anesthetized with 10％ chloral hydrate 350 mg/kg.Normal saline 0.8 ml was injected via the caudal vein in and CLP groups,while 10％ methylene blue 15 mg/kg (in normal saline 0.8 ml) was injected via the caudal vein in group MB.Sepsis was induced by CLP after the end of administration in CLP and MB groups.Twenty animals in each group were chosen and observed for 72 h survival rate.Ten animals were sacrificed in each group at 18 h after operation and kidney specimens were removed for microscopic examination and for determination of the expression of poly (ADP-ribose) polymerase (PARP-1) using immunohistochemistry and Western blot.Blood samples were taken from the heart for determination of serum concentrations of blood urea nitrogen (BUN),creatinine (Cr),cystatin C and neutrophil gelatinase-associated lipocalin (NGAL).Results Compared with group S,the survival rate was significantly decreased at 24,48 and 72 h after operation,and the serum concentrations of BUN,Cr,cystatin C and NGAL and expression of PARP-1 in kidney tissues were increased in CLP and MB groups (P ＜ 0.05).Compared with group CLP,the survival rate was significantly increased at 24 and 48 h after operation,and the serum concentrations of BUN,Cr,cystatin C and NGAL and expression of PARP-1 in kidney tissues were decreased in group MB (P ＜ 0.05).The pathological changes were significantly attenuated in group MB as compared with group CLP.Conclusion Methylene blue pertreatment can attenuate acute kidney injury induced by sepsis in rats through down-regulating the expression of PARP-1.

Objective To study the significance of expression of c-met protein in gastric cancer tissue and to explore the correlation between c-met and micro-vessel density (MVD). Methods c-met protein and MVD in gastric cancerous tissue were determined by immunohistochemistry in 47 patients. Results The positive rate of c-met in gastric tissue was 85.1%. The expression of c-met was much higher in patients with tumor diameter larger than 5cm, with deeper invasion, regional lymph nodes metastasis, lymph vessel and venous invasion, and TNM stage Ⅲ-Ⅳ (P

Radix bupleuri is mainly used to treat cold fever and liver disease, and the curative effect is remarkable. The prepara-tions previously explored including bupleurum injection, tablets and capsules all show their own characters and different applicable peo-ple. Recently, many researchers carried out extensive research on its dosage forms. The early formulations have been improved as well, and successively developed dropping pill, nasal sprays, transdermal patches and the other new dosage forms. In the paper, the recent research progress in bupieurum preparations were summarized in order to provide reference for the improvement and application.

Objective: To optimize the extraction conditions of semi-bionic extraction for Gardenia jasminoides Ellis. Methods:The best extraction conditions of the semi-bionic extraction for Gardenia jasminoides Ellis was screened by uniform design with yield of geniposide, total iridoid glycosides and dry extract as the indices. Results:The optimal conditions were as follows:the pH value of wa-ter for the lst, 2nd and 3rd decoction was 2. 0, 6. 5 and 9. 0, respectively, and the extraction time was 2. 0, 1. 0 and 1. 0h according-ly. Conclusion:The optimized extraction conditions by uniform design is scientific and reasonable.

Objective To investigate the expression and significance of Claudin 3 in adenocarcinoma of esophagus and gastric cardia.Methods We subjected pathologically confirmed 16 cases of esophageal adenocarcinoma(EAC),20 cases of esophageal squamous cell carcinoma(ESC),and 40 cases of gastric cardiac adenocarcinoma(CAC,including 15 diffuse and 25 intestinal cases).Tissue samples were collected with aid of gastroscope.Fifteen samples of normal esophageal mucosa and 15 samples of normal cardiac mucosa served as control.RT-PCR and Western blotting were used respectively to detect the mRNA and protein expression of Claudin 3 in these tissue samples.Results The mRNA and protein expressions of Claudin 3 were increased in EAC and intestinal CA compared with those in normal tissues,ESC and diffuse CAC.There was no significant difference of the expression of Claudin 3 between normal group and ESC and diffuse CAC group.Conclusion Claudin 3 might be related to the development of EAC and intestinal CAC,and expected to be the tumor maker and therapeutic target of this kind of carcinoma.

Objective: To study the UPLC-MS fingerprint of Bupleurum marginatum DC. in northwest Hubei and establish the quality evaluation system for the herb. Methods:A UPLC-MS method was used to analyze 10 samples of B. marginatum DC with the following chromatographic conditions:an ACQUITY UPLC? RBEH C18 column (100 mm × 2. 1 mm, 1. 7 μm) was used, the mobile phase consisted of acetonitrile-0. 3% formic acid water with gradient elution, the flow rate was 0. 2 ml·min-1, and the detection wavelength was 203nm with ESI(-). Results:There were 8 common peaks in the UPLC-MS fingerprint of B. marginatum DC. Through analyzing the information of mass spectrometry and combining the references, 6 peaks were identified. Conclusion:The UPLC-MS fin-gerprint method is simple,rapid and feasible. The acquired UPLC-MS fingerprint of B. marginatum DC. and the evaluation indices can provide the scientific quality assessment of B. marginatum DC.

Objective:To establish the HPLC fingerprints of Atractylodes lancea in northwest of Hubei province to provide the basis for the quality control and variety identification of Atractylodes lancea. Methods:The fingerprints were detected by HPLC. The separa-tion was achieved on a Diamonsil 2# C18(250 mm ×4.6 mm,5 μm)chromatographic column at 30℃ using acetonitrile-water (gradient elution) as the mobile phase at a flow rate of 1. 0 ml·min-1 . The detection wavelength was 340nm. Results: There were 13 common peaks in the HPLC fingerprints of 14 batches of Atractylodes lancea with promising separation. According to the results of clustering a-nalysis,Atractylodes lancea in northwest of Hubei province was clustered into two categories. Conclusion:A simple, feasible and relia-ble method is provided for the quality control of Atractylodes lancea.

0 05). APC mutation was correlated with intestinal type of gastric cancer ( P 0 05). APC mutation was detected in all MSI Low or MSS, while no mutation was found in MSI H. Conclusion hMLH1 mutation and promoter methylation are involved in MSI pathway and APC mutation is involved in LOH pathway in gastric cancer. H pylori infection is not related to hMLH1, APC mutation and promoter methylation in gastric cancer.

Objective To detect the differential expression genes(DEGs)between Barrettg esophagus(BE)and normal esophagus with oligomicroarray,and to explore the target genes related to the development of BE.Methods The total RNAs of matched BE and normal esophagus mucosa from saIne patient were isolated with one step Trizol method.Matched RNAs were qualified with 10g/L agarose gel electrophoresis.After tRNA purification,cRNAs were synthesized and labeled with fluorescence.which were tIlen hybridized with Agilent oligomicroarray containing 30,968 probes.The fluorescence intensity features were detected by Agilent scanner and quantified by software Feature Extraction.Results On average,2 biopsies by disposable jumbo biopsy forceps provided approximately 5μg RNA required for microarray.The total RNA,reverse transcription product and fluorescence labeled cRNA were all of high quality.Among 2-fold DEGs,there were 142 up-regulated genes and 284 down-regulated genes including 15 bel-2 related genes such as bel-2,MCL1,BAX,BIK and BCLAF1 Conclusion Microarray-based studies are feasible in endoscopically obtained tissues.The development of BE is a complicated process involving multi-genes,in which abnormal expression of bel-2 family related genes might be involved,but the exact mechanism needs further research.

BACKGROUND: Microsatellite instability (MSI), an important gene change type, plays animportant role in the occurrence of tumor. Mismatch repair gene induces its occurrence. Although the effect of mismatch repair gene hMLH1 mutation in the hereditary nonpolyposis colorectal cancers (HNPCC) has been reported, its effect on the sporadic colorectal carcinoma lacks in-depth study.OBJECTIVE: To investigate the effect of mismatch repair gene hMLH1 mutation on colorectal carcinogenesis, and its correlation with MSI.DESIGN: Single-sample experiment.SETTING: Department of Gastroenterology, Southwest Hospital of Third Military Medical University of Chinese PLA.PARTICIPANTS: Seventy-six cases of sporadic colorectal carcinoma and corresponding normal tissues were obtained from surgically resected specimens of coloreetal carcinoma in Southwest Hospital between January 2001and December 2003. No patients had family history of tumor, or had received radiotherapy and chemotherapy. Patients were informed of the experiment.METHODS: Mutation of hMLH1 was detected by two-dimensional electrophoresis and DNA sequencing; MSI was analyzed by PCR-based methods.MAIN OUTCOME MEASURES: ① Detection rate of hMLH1 mutation of colorectal carcinoma and MSI. ② The relationship of MSI and hMLH1 mutation.RESULTS: Seventy-six cases of sporadic colorectal carcinoma were studied for hMLH1 mutation and MSI. hMLH1 mutation was detected in 8 (10.5%) cases of colorectal carcinomas while MSI was detected in 20 (26.3%) cases of colorectal carcinomas. Frequency of hMLH1 mutation and MSI was significantly higher in right colorectal cancer than in left colorec tal cancer (6/26 vs 2/50, x2=4.739, P=0.029; 11/26 vs 9/50,x2=5.212,P=0.022). No association was observed between hMLH1 mutation or MSI and tumor size, differentiation, histological type, depth of invasion, metastasis or clinical pathological stages. ② MSI was divided into high-frequency group (≥ 2 loci, n=10) and low-frequency group (1 locus, n-10), and MSI negative group (n=56). 8 hMLH1 mutations were all detected in high frequency MSI group, but no mutation was found in low frequency MSI or MSI negative groups.CONCLUSION: hMLH1 mutation and MSI occur in cancer of the right large intestine and hMLH1 mutation is involved in carcinogenesis of some sporadic colorectal cancer with high-frequency MSI.