Objective To introduce the pattern of intestinal parasites infection in Guyana,South America.Methods Direct faecal smears were performed by using 1% Lugol′s iodine wet-mount preparations.Results The infection rate of intestinal parasites in faeces was(6.54%) in 5,200 patients.The intestinal parasites found most commonly were:Hookworm 2.33%(121/5,200);G.lamblia 1.65%(86/5,200);Ascarislumbricoides 0.75%(39/5,200);Whipworm 0.69%(36/5,200);E.histolytica 0.42%(22/5,200).Conclusions Infection rate of intestinal parasites is quite high in the citizen of Georgetown district.

PURPOSE To investigate the effects of all trans retinoic acid(ATRA) on the growth of gastric cancer cells and explore the mechanism relevant to enzyme.METHODS With MTT method the growth inhibition of gastric cancer cells was measured;the activity of lactate dehydrogenase(LDH),alkaline phosphatase(ALP) and ? glucuronidase(? G) was measured with metaplate;The activity of succinate dehydrogenase(SDH) was assayed with cytochemical method.RESULTS The growth of MGc80 3, but not MKN 45, was inhibited,which is dependent on the concentration of ATRA.The activity of LDH,ALP and ? G of Mgc80 3 was suppressed and the activity of SDH of Mgc80 3 was enhanced.But all the four enzymes′ activity of MKN 45 were not obviously changed except that introcelluar ? G activity was inhibited.CONCLUSIONS The sensitivity of gastric cancer cells to ATRA is different,which is closely related with the change of enzyme activity.

Objective To explore the value of transthoracic echocardiography(TTE) and 64-slice spiral computed tomography(CT) in the diagnosis of complex congenital heart disease(CHD). Methods Ninety-seven patients diagnosed as CHD by TTE underwent 64-slice spiral CT for cardiovascular examination. The results were compared with the results by cardiovascular angiography and from surgery. Results The total diagnosis accordance rated by TEE was 90. 2% and that by spiral CT was 92.5%. There was no difference in diagnosis accuracy rate between TTE and spiral CT. The diagnosis accuracy rate in intracardiae defomities by TTE was 99.2 %, higher than 87.50% by spiral CT. However, the diagnosis accuracy rate in extracardiac defomities and ventricular-arterial connections was 99. 0% by spiral CT, higher than 78. 6% by TTE. The combination of TTE and spiral CT can raise the diagnosis accuracy rate to 99. 1%. Conclusions TTE is of significant value in complex CHD diagnosis,especially in the diagnosis of intracardiac defomities. The combination of TTE and spiral CT can raise the diagnosis accuracy rate of various kinds of complex CHD.

Objective: To design and confirm the cleavage activity of ribozyme Rz217 to oncogene ki-rasG12V messenger RNA and search for a new method for gene therapy targeting oncogene ki-ras. Methods: According to Symon' s principle,design an ribozyme specific for ki-rasc12v mRNA, both the constructs for transcription in vitro of ribozyme Rz217 and ki-ras exonl and the mammalian expression constructs of ribozyme Rz217 were constructed by DNA recombinant technique,ribozyme Rz217 and ki-ras exonl mRNA was obtained by transcription in vitro with T7 and SP6 RNA polymerase. Pancre atic carcinoma cell line PaTu8988 and human hepatocellular carcinomacell line BEL7404 were transfected with Rz217 mammalian expression constructs and the level of endogenous ki-rasG12V mRNA or ki-ras mRNA was determined by semiquantitative RT-PCR. Results: Not only in vitro but also in vivo, ribozyme Rz217 can cleave the mRNA of oncogene ki-ras (G12V) in site-specific manner and can not cleave the mRNA of wild-type ki-ras. Conclusion: Ribozyme Rz217 can cleave oncogene ki-rasc12v mRNA and the cleveage is specific for ki-rasG12V mRNA.

BACKGROUND:Previous studies have verified that mesenchymal stem cells could be transplanted into inflammatory bowel mucosa to repair inflammatory bowel tissue. OBJECTIVE:To observe the differential gene expression in large intestine before and after mesenchymal stem celltransplantation in repair of inflammatory bowel tissue of rats using microarray technology, and to primarily discover the main genes during mesenchymal stem celltransplantation, differentiation, and reparation in inflammatory colorectal tissue region. METHODS:Healthy Sprague-Dawley rats were randomly divided into two groups. Experimental rat models of inflammatory bowel disease were established using trinitrobenzene sulfonic acid via enema. At 24 hours after model establishment, green fluorescent protein-labeled mesenchymal stem cells were infused via the caudal vein. The control group was treated with physiological saline by enema, instead of trinitrobenzene sulfonic acid. At 28 days, large intestine was obtained from the experimental group and control group. Differential y expressed genes were screened in the experimental and control groups using microarray technique. RESULTS AND CONCLUSION:The microarray analysis results showed that there were 388 differential genes in the control and experimental groups (P<0.05, FC>2), in which 191 were up-expressed, and 197 were down-expressed. Al of these genes were mainly involved in inflammatory reaction, immune reaction and celldifferentiation. In the top 10 up-regulation and down-regulation differential genes (total y 20 genes), 3 genes were involved in inflammation, 3 genes were involved in immune reaction, and 2 genes were related to stem celldifferentiation. In the 388 genes, 33 were related to signaling pathways (P<0.05), 6 related to inflammation, 8 related to immunity, and 5 related to stem celldifferentiation. Results suggested that the main genes involved in mesenchymal stem cells in repair of inflammatory bowel tissue were primarily screened using gene expression microarray technique.

BACKGROUND:β-catenin is the most critical signaling molecule in the Wnt/β-catenin signaling pathway, which is involved in the regulation of cellproliferation, differentiation and tissue self-healing balance.
OBJECTIVE:To construct a stableβ-catenin over-expression lentivirus-mediated vector and to transfect mesenchymal stem cells line for investigating its effects on proliferation and migration of mesenchymal stem cells.
METHODS:Over-expression vector, PLV-EF1A-catenin-RFP, was constructed and transfected the 293T cellto infect mesenchymal stem cells, and positive cells were selected with puromycin. The up-regulated efficiency of targetingβ-catenin gene at mRNA level was detected by real-time quantitative PCR, the effect on proliferation of mesenchmal stem cellwas assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and growth curve, and the migration ability was detected by Transwel motility assay.
RESULTS AND CONCLUSION:The lentiviral vector targetingβ-catenin gene was constructed successful y, and a stable mesenchymal stem cellline that up-regulatedβ-catenin was established. Real-time quantitative PCR results showed that the expression ofβ-catenin gene was efficiently up-regulated by infecting PLV-EF1A-catenin-RFP (P<0.05). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and growth curve showed that celldoubling time was shortened after infected with pLV-EF1A-catenin-RFP (P<0.05), indicating that the over-expression of theβ-catenin gene successful y increased the proliferative capability of mesenchymal stem cells. The Transwel assay also showed similar increasing results on the migration ability (P<0.01). The lenvivirus-mediated over-expression of theβ-catenin gene can be used to increase the proliferation and migration abilities of the mesenchymal stem cells.

Objective To explore the operation procudure and notice of establishing a rat model of abdominal heterotopic heart transplantation by single operator,and summarize the detailed skill and experience for beginners.Methods 68 pairs male SD(recipients)/Wistar(donors) rats,preoperative anesthesia for recipients and donors at the same time,8/0 line was preparing for blocking recipients vena cava and abdominal aortic,blocking the branch vessel.Left and right superior vena ligature respectively with hilar,cutting the ascending aorta,perfusion cardioplegia,free pulmonary artery,across transverse sinus of pericardium for pulmonary artery cutting,atrial wash,free donor heart.Choosed appropriate incision of receptor abdominal blood vessels,mattress suture at 6 and 12 points,a single suture for vent gases,continuous suture in artery,single needle in vein,reperfusion after exhaust gas.Recording operation time,HE staining,IL-1β/CD3 immunohistochemistry slice,flow cytometry analysis of CD4+T/CD8+T lymphocytes subsets in the peripheral blood.Results Sixty-eight cases were treated by single operater,the operation time was(58.8±4.5)minutes,artery suture time was(7.6±2.2)minutes,vein suture time was(13.5±4.2)minutes,the total donor heart ischemic time was(31.8±4.5)minutes,the success rate was 88.2％.The rejection reaction was stronger on the third and fifth day after surgery,with high expression of IL-1β/CD3 in cardiac allografts.The CD4+T/CD8+T lymphocytes subsets increased in the peripheral blood at first day after heart transplantation.Conclusion By fully preparation and skillfully operation,establishing a rat model of abdominal heterotopic heart transplantation by single operator has a stable success rate.

Objective:To optimize the extraction process for the total alkaloids of Longzuan Tongbi Fang by central composite design and response surface method.Methods:The independent variables were the ethanol contention,solvent consumption and extraction time,and the dependent variable was the weighted overall desirability of the content of total alkaloids and kukoline.The extraction process for the total alkaloids of Longzuan Tongbi Fang was optimized by central composite design and response surface method.Results:The optimum extraction conditions were as follows:9-fold amount of solvent was used,refluxed for 130 min,extracted twice,and the ethanol concentration was 75%.Conclusion:The central composite design and response surface method used for the extraction optimization for the total alkaloids of Longzuan Tongbi Fang is very simple and highly predictive.

An approach was proposed to evaluate preparation technology by means of fingerprint-peak matching technology of high performance liquid chromatography with diode array detector (HPLC-DAD).Similarity and hierarchical clustering analysis (HCA) were applied to identify the 15 batches of Xiaochaihu granules from different manufacturers and our laboratory,and peak pattern matching between the composite formulae and Radix Bupleuri Chinensis,which was one of the main ingredients of Xiaochaihu granules,was utilized to evaluate the preparation technology of Xiaochaihu granules via the indexes of the relative deviation of retention time (RT) and UV spectrum feature similarity of their corresponding peaks.Eleven matching peaks were found between Xiaochalhu granules and Radix Bupleuri Chinensis.However,the saikosaponin A and saikosaponin D,which are the important active components in Radix Bupleuri Chinensis,were not found in Xiaochaihu granules from any manufacturers.The peak areas of 11 characteristic peaks of Xiaochaihu granules samples formed a matrix of 11 × 15.The result of HCA showed that Xiaochaihu granules samples were divided into four kinds of category.Xiaochaihu granules samples from the same manufacturer were basically clustered of the same category.The results suggested that the saikosaponin A and saikosaponin D are prone to structural transformation under the condition of decoction and in the presence of the organic acidic components.These active components,existing in raw herb,might transform to a series of non-active secondary saikosaponin due to unfavourable preparation technology.So the conventional decoction-based preparation technology of Xiaochaihu granules might greatly affect its quality and therapeutic effectiveness. This study demonstrates that fingerprint-peak matching technology can not only be used for quality control of this composite formulae,but also provide some guidance for preparation technology of Xiaochaihu granules.

Aim ToexploretheeffectsofnewdrugT-006 on improving learning and memory abilities in scopolamine-induced dementia mice and its possible mechanism.Methods 72maleKunmingmicewere randomly divided into six groups:normal control group,model group,donepezil treatment group,T -006 treatment group with different doses(1,3 and 10 mg·kg-1 ).All mice were treated by intragastric ad-ministration for 14 consecutive days. Learning and memory abilities were tested by a five-day Morris water maze trial from the 1 1 th day.the first 4 days of the five-day Morris water maze,the navigation test was performed,the last day of Morris water maze is the spatial probe test.During the navigation test, mice were intraperitoneally given 2 mg · kg-1 scopolamine 20 minutes before entering the water,while normal control group mice administrated with sterile saline in-stead.Mice were not given T-006 nor scopolamine in spatial probe test.After Morris water maze,all mice were sacrificed for hippocampus and cortex.The activi-ties of AchE and SOD and the levels of GSH and MDA in hippocampus and cortex were measured after tissue harvesting.Results Comparedwithmodelgroup,T-006 could obviously improve learning and memory abil-ities in scopolamine-induced mice, significantly in-crease the levels of SOD and GSH and decrease the levelsofMDAandAchE.Conclusion T-006can significantly improve cognitive abilities in scopolamine-induced dementia mice,and its relevant mechanism may be closely related to its antioxidative effect and the ability to decrease AchE level.