Diabetic gastroparesis (DGP) is a common complication in patients with diabetes mellitus of long duration, presenting with recurrent nausea, vomiting, postprandial fullness and early satiety caused by delayed gastric emptying. The treatments of DGP include dietary therapy, nutritional support, glycemic control, use of prokinetic and antiemetic agents. This review focuses on current status of the drug treatment and the progress of new agents of DGF under the preclinical and clinical trials.

Objective To explore the association of the Arg64 polymorphism β-3-adrenergic receptor(β3-AR) gene with polycystic ovary syndrome(PCOS)and its characterization with obesity,insulin resistance(IR)，blood pressure (BP), blood lipid spectrum. Methods The polymorphism of the β-3-AR gene was determined in 163 patients with PCOS and 100 controls. Body mass index(BMI), waist to hip circumference ratio(WHR), BP were examined. Blood glucose,blood lipid spectrum,insulin were also measured. Insulin resistance index was calculated. Results The frequency of Arg64 allele in PCOS women was significantly higher than that of controls(0.33 vs 0.13 respectively, P<0.05). Fasting insulin, insulin resistance index, WHR in Arg64 group were significantly higher than those of Trp64 group. The frequency of Arg64 allele was positively correlated with FINS, insulin resistance index, WHR. Conclusion The Arg64 mutation in the β-3-adrenergic receptor(β-3-AR) gene might contribute to development of PCOS and IR.

This paper is to report the development of a rapid and sensitive method for the determination of s-oleylpropanolamide (OPA) in various tissues of rat (brain, heart, lung, liver, spleen, small intestine, kidney, adipose tissue and muscle), and to assess the applicability of the assay to tissue distribution. OPA was extracted by liquid-liquid extraction method with undecylenoylethanolamide as an internal standard. The concentrations of OPA were determined by LC-MS/MS after a single intragastric dose of 50 mg x kg(-1) at 4 time points (5 rats per group). With multiple reactions monitoring mode (MRM) the limit of quantification (LLOQ) was determined at 1 microg x L(-1). The calibration curve was linear from 1 to 2 x 104 microg x L(-1) (r > or = 0.999 0) for tissue homogenates. Validation parameters such as accuracy, precision and recovery were found to be within the acceptance criteria of the assay validation guidelines. The highest concentration was found in small intestine (the highest time point is 15 min) and heart (the highest time point is 90 min). The assay is rapid, sensitive and applicable to studying tissue distribution of OPA in rats.

Objective To establish a liquid chromatography-tandem mass spectrometry ( LC-MS/MS) method for the quantification of creatinine-correctedsarcosine in urine for the prostate cancer diagnosis and treatment.Methods It performed the method establishment and evaluation in this study.Random unrine samples were collected from 36 subjects with prostate cancer, 15 subjects with benign prostatic hyperplasia and 76 healthy people receiving medical examination.Urine samples mixed with [ 2 H3 ]-labeled sarcosine were treated by precolumn derivation using dansyl chloride, then analyzed by LC-MS/MSsystem in multiple reaction monitor ( MRM) mode.Sarcosine and creatinine were quantified by the isotope internal standard method and the standard curve was employed with a series of calibration.The limit of detection, precision and recovery were also evaluated in this study.The results of this methodology were compared with those of the enzymatic method.Results Sarcosine could be distinguished against its isomers completely. The linear equation of sarcosine was Y=2.045 6X+0.068 9, R2 =0.994.The limit of detection and limit of quantity were 8 ng/ml and 25 ng/ml respectively.The intraassay and interassay coefficients of variation were both below 6%.The recovery ratio of sarcosine ranged from 96.8%to 105.1%.The results from the ID-LC-MS method correlated with those from enzymatic method (R2 =0.815, P <0.01).Compared to enzymatic method, the average bias of sarcosine was -37.1%.Conclusions It established a LC-MS method for urinary sarcosine quantification with good specificity, sensitivity and repeatability.This method can provide a reliable platform for the diagnosis of prostate cancer.

BACKGROUND:There is no clear conclusion on whether the lung cancer stem cels can induce to spheroid formation and have tumorigenicity. OBJECTIVE: To observe the spheroid formation induced by human lung adenocarcinoma cel line SPC-A1 and the tumorigenic ability of lung cancer stem cels. METHODS:SPC-A1 at proliferating phase was cultured in serum-free DF12 culture medium, and then recombinant human insulin-like growth factor-1, recombinant human epidermal growth factor, and recombinant human fibroblast growth factor-10 were added to induce spheroid cels. Immune fluorescence detection and PCR amplification were done to understand the expression of stem cel associated markers. NOD-SCID immunodeficient mice were subcutaneously implanted with lung spheroid cels to observe the tumor growth.In vivo fluorescence imager was used for radiography. RESULTS AND CONCLUSION:After 5-10 days, lung spheroid cels were harvested. RT-PCR results showed that lung spheroid cels were positive for CD24, CD221, CCSP and SP-C. In addition, the lung spheroid cels and purified CD24+, CD221+ lung cancer stem cels were both positive for TTF-1 of lung stem cels, OCT4 and Nanog of embryonic stem cels and TTF-1 of Bmi-1 lung stem cels. The fluorescence detection showed that over 80% lung spheroid cels expressed CCSP and OCT4; SPC-A1 cels had the characteristics of alveolar type II cels, and also expressed SP-C protein, but only about 5% of the cels expressed CCSP and OCT4. At 50 days after subcutaneous implantation of lung spheroid cels, in vivo fluorescence imaging showed that the diameter of tumor in mice was 1 cm, indicating human lung adenocarcinoma cel line SPC-A1 can induce the spheroid formation, and lung cancer stem cels rich in the cel spheres have the tumorigenic ability.

Objective To explore the effects of up-and down-regulation of Cox-2 expression on the growth and radiation sensitivity of human esophageal cancer EC9706 xenograft in nude mice.Methods Cox-2 specific siRNA and Cox-2 gene eukaryotic expression vector were constructed and transfected to esophageal cancer cells EC9706, and the stable transfected cell lines were obtained by the method of G418 screening.The expressions of Cox-2 mRNA and its protein were detected by RT-PCR and Western blot, respectively.The inhibitory effects of Cox-2 regulation combined with X-ray irradiation on cancer cell growth were detected by the nude mouse xenograft assay.Results Cox-2 gene expression was significantly decreased in the Cox-2 down-regulated group and increased in the Cox-2 up-regulated group.Compared with the control group without gene transfer, the average volumes of EC9706 xenograft tumor in the Cox-2 up-and down-regulated group significantly decreased (F =34.26, P ＜ 0.05) and increased (F =26.38, P ＜ 0.05) , respectively.After 20 Gy X-ray irradiation, the average volume of xenograft was significantly reduced in the Cox-2 down-regulated group (F =16.35, P ＜ 0.05) , but it had no significantly changes in the Cox-2 up-regulated group.Conclusions Down-regulation of Cox-2 expression inhibits the growth of human esophageal cancer EC9706 xenograft in a nude mice and increases cell radiation sensitivity, but upregulation of Cox-2 expression makes tumor cells to become radioresistant.

Objective To investigate the expression level of LMO1 in gastric cancer tissues and human gastric cancer cell lines,and explore the invasive and metastatic potential of LMO1 gene silencing by small interfering RNA on the human gastric cancer cell line MKN28.Methods Immunohistochemical technique was applied to detect the expression of LOM1 protein in gastric cancer tissues and tumor adjacent tissues of paraffin specimens in 30 cases.The expression levels of LMO1 in human gastric cancer cell lines AGS,BGC-823,SGC-7901,MKN28 and human gastric mucosal epithelial cells GES were detected by realtime-PCR and Western blot.Using LipofectamineTM 2000,LMO1 siRNA was transfected into MKN28 cellsin vitro (siRNA transfect group).Negative control was established.Real time-PCR and Western blot was used to examine the difference of LMO1 expression.Transwell assays were performed to identify the differences and changes of invasive and metastatic ability in gastric cancer cell line MKN28.Western blot was used to examine the expression levels of E-cadherin,MMP-9 and VEGF.Results Positive rate of LOM1 protein in gastric cancer tissues(77％)was higher than that in tumor adjacent tissues (17％) (x2 =21.70,P ＜ 0.01).Positive rate of Vavl protein was higher in lymphatic metastasis group than in non-lymphatic metastasis group(x2 =5.83,P =0.02).Compared with GES,the expression level of LMO1 increased significantly in gastric cancer cell lines,especially in MKN28 (P ＜ 0.01).The expression levels of LMO1 mRNA and protein in LMO1-siRNA transfected MKN28 cells were lower than the matched negative control cells (P ＜0.01).The invasive and metastatic potentials of LMO1-siRNA transfected MKN28 cellssignificantly decreased (t =-11.53,P ＜0.01;t =-10.68,P ＜0.01).The expression levels of E-cadherin were higher than the matched negative control cells;and MMP-9,VEGF protein in LMO1-siRNA transfected MKN28 cells were lower than the matched negative control cells (P ＜ 0.01).Conclusions LMO1 has higher expression level in gastric cancer tissues and some gastric cancer cell lines,and down-regulation of LMO1 can inhibit the invasion and metastasis ability of gastric cancer.

OBJECTIVE:To observe the efficacy and safety of paclitaxel combined with cisplatin and fluorouracil for gastric cancer with liver metastases via indwelling hepatic arterial catheter. METHODS:56 gastric cancer patients with liver metastases were randomly divided into control group(28 cases)and observation group(28 cases). Control group received Paclitaxel injection 135 mg/m2,d1+Cisplatin injection 75 mg/m2,d1+Fluorouracil injection 750 mg/m2,d1-5,pumping administrated via central venous. Ob-servation group received Paclitaxel injection 135 mg/m2,d1+Cisplatin injection 75 mg/m2,d1+Fluorouracil injection 750 mg/m2,d1-5, administrated via indwelling hepatic arterial catheter. 3-4 weeks were a course,it lasted 8 courses at most. Magnesium isoglycyrrhiz-inate injection 200 mg/d was intravenously infused for liver protection in 2 groups during treatment. Clinical efficacy,serum car-cinoembryonic antigen (CEA),alanine aminotransferase (ALT),aspartate aminotransferase (AST) levels before and after treat-ment,and the incidence of adverse reactions in 2 groups observed. RESULTS:Short-term clinical efficacy in observation group was significantly higher than control group,with statistical significance(P<0.01). Before treatment,there were no significant dif-ferences in CEA,ALT and AST levels(P>0.05). After treatment,CEA,ALT and AST levels in 2 groups were significantly high-er than before,ALT and AST levels in observation group were significantly higher than control group,while CEA level in observa-tion group was lower than control group,with statistical significances(P<0.05). The incidences of bone marrow suppression,nau-sea,vomiting and fever in observation group were significantly lower than control group,with statistical significances (P<0.05). CONCLUSIONS:Paclitaxel combined with cisplatin and fluorouracil has good efficacy for gastric cancer with liver metastases via indwelling hepatic arterial catheter,while it exists liver dysfunction.

Objective To provide a simple, convenient, and safe anesthesia method for the establishment of a M1 segment of middle cerebral artery occlusion model in rhesus monkey or other large laboratory animals.Method Twenty male rhesus monkeys weighing 7-11 kg (ages 7-9 years) from Academy of Military Medical Sciences were used in this study.Sumianxin injection combined with 0.1 mg/kg ketamine was given before endotracheal intubation (ID:4.5-5.5#).The animals were then transported to an interventional operation room, where the intravenous access was established and a urinary catheter was inserted into the urinary bladder.Mechanical ventilation was used during the surgery, propofol was continuously injected in a speed of 2-4 mg/kg/h, and Sumianxin-ketamine could be given if necessary to maintain adequate anesthesia depth.The dose was adjusted according to vital signs of the rhesus such as body movements, physiological parameters, and demand of surgery.Brain MRI examination was performed before and after thrombolysis.Anesthetic injection was suspended and the animals were allowed to have a spontaneous breathing every time before the MRI text.Heart rates, temperature, non-invasive blood pressure, and SpO2 were monitored during the whole surgery.Blood samples were taken from the radial artery for blood gas analysis after anesthesia induction and during operation.Results All the 20 animals underwent the operation successfully, no animal had restlessness, respiratory depression, arrhythmia and other serious complications.At the end of the study, animals awake soon.Fifteen of them survived longer than 24 hours, only 5 died from serious cerebral hemorrhage and larger cerebral infarction.Conclusions General endotracheal anesthesia is safe for rhesus monkeys during such interventional operation and MRI examination.