BACKGROUND:Preterm premature rupture of the membrane is a frequent complication during gestational period. Three-dimensional culture can help damaged tissue restore anatomical integrality,being worth for clinical application. OBJECTIVE:To explore biological characteristics of human amniotic cells three-dimensionally cultured by fibrin scaffold,study the possibility of cell/scaffold composite material for repairing preterm premature rupture of the membrane,and to compare with normal monolayer culture. DESIGN,TIME AND SETTING:An in vitro cell/scaffold study was performed at the Basic Institute of Chongqing University of Medical Science between December 2007 and November 2008. MATERIALS:Amnion tissue derived from uterine-incision delivery women was provided by the Department of Obstetrics and Gynecology,the First Affiliated Hospital,Chongqing University of Medical Sciences;fibrinogen was provided by Sigma,USA. METHODS:Either epithelial cells or mesenchymal cells,being isolated from human amniotic membrane,were cultivated using enzyme digestion and repeated adherence methods. Fibrinogen was conglomerated,and epithelial cells of logarithmic growth were plated onto the surface of fibrin scaffold to simulate a three-dimensional culture in vitro. Before conglomerating fibrinogen,mesenchymal cells of logarithmic growth were mixed with fibrin solution to simulate a three-dimensional culture in vivo. In addition,either epithelial cells or mesenchymal cells were incubated onto 24-well plate for normal monolayer culture. MAIN OUTCOME MEASURES:Cell morphology using inverted microscopy,proliferation of mesenchymal cells,and interaction between mesenchymal cells and scaffold under different culture environments. RESULTS:Epithelial cells were round and smooth on the surface of fibrin scaffold,pseudopodia were stretched out,and microvillus were rich. Mesenchymal cells in fibrin glue were fusiform in shape and stretched out along scaffold. Cells formed net structure at different surfaces. By three-dimensional culture,proliferation of mesenchymal cells was stable but slow compared to monolayer culture. By in vivo three-dimensional culture,fibrin glue gradually shrank and gel thickness gradually decreased. On the 5th day,the gel thickness was only 40% for the initial thickness,and on the 15th day,gel thickness was 10%. CONCLUSION:Both epithelial cells and mesenchymal cells can sterically grow on fibrin scaffold,and the proliferation is stable. Fibrin glue gradually shrinks in cell/fibrin composite scaffold,suggesting that amniotic cells/fibrin composite scaffold can simulate body tissues to repair preterm premature rupture of the membrane.

ObjectiveTo investigate the anti tumor effect of Giant Typhonium Rhizome.Methods SH-SY5Y was treated for 24 or 48 h by Giant Typhonium Rhizome.Cell Counting Kit-8 (CCK-8) cell proliferation and cytotoxicity assays were applied to detect the inhibition of SH-SY5Y cells treated with aqueous extract of white monkshood root.AV/PI double staining flow cytometry was applied to explore the mechanism of growth inhibition.ResultsSH-SY5Y cells could be inhibited by root aqueous extract of Giant Typhonium Rhizome,the IC50 at 48 h (50％ inhibitory concentration) was 0.862 mg/ml,while the IC50 at 24 h was 1.158 mg/ml.Western-blot results showed that root aqueous extract of Giant Typhonium Rhizome could induce apoptosis to inhibit tumor cells.ConclusionThe Rhizoma typhonii inhibit the growth of SH-SY5Y.

ObjectiveTo investigate whether the reactive oxygen species involved in inhibiting proliferation of human bone marrow cells of neuroblastoma cells (SH-SY5Y)process by giant typhonium rhizome water extract,and to investigate the initial mechanism of anti-tumor effect of giant typhonium rhizome in vitro.MethodsAfter giant typhonium rhizome water extract processing of human bone marrow neuroblastoma cells (SH-SY5Y)at 24 h and 48 h,the cells were collected,and western blotting reaction methods such as flow cytometry was used to study the levels of ROS and SH-SY5Y apoptosis.ResultsThe levels of ROS elevated from before dosing (4.9±1.6)％ to (39.7 ± 11.8)％ in the process of giant typhonium rhizome water extract induced apoptosis in SH-SY5Y; after ROS inhibition was added,SH-SY5Y apoptosis level induced by giant typhonium rhizome water extract had decreased.ConclusionROS involved in the process of giant typhonium rhizome water extract inhibiting cell proliferation in SH-SY5Y.

Objective To observe influence of Benazepril combined with Jinshuibao capsules on urine protein and Ccr in patients hypertensive renal disease. Methods 47 hypertensive renal disease patients were randomly divided into control group and combine group. Patients in control group were treated with Benazepril, while patients in combined group were treated Jinshuibao plus Benazepril. Clinic index change after treatment for 9 weeks was observed, and calculation endogenous creatinine clearance rate was calculated. Results After treatment for 9 weeks urine protein in 24 hours of two groups were both lower than before( t1 = 1. 9997 ,t2 = 3.3088 ,P ＜ 0. 05 or P ＜ 0. 01 ),while the endogenous creatinine clearance rate were both higher than before ( t1 = 1. 3990, t2 = 1. 4442, P ＞ 0. 05 ), and the press were all in improvment ( t1 = 2.0047, t2 = 2.4557, t3 = 2.0426, t4 = 2.4026, P ＜ 0. 05 ); urine protein in 24 hours of combine group was lower than control group( t = 2.0015,P ＜ 0. 05) Conclusion Benazepril plus Jinshuibao capsules could obviously abate urine protein on patients with hypertensive disease and hypertensive renal disease

Objective To study CT feature of hypoxic-ischemic encephapathy in neonates and its value in predicting prognosis.Methods CT scanning was performed in 62 cases with HIE.CT feafure were analyzed.Results Of 62 cases during their neonatal period,brain edema was seen in 38 the lower density's lesions of subcortical with matter in 42,cerebral hemorrhage in 8,subarachnoid space hemorrhage in 16,SDH in 4,and intraventricular hemorrhage in 2.Of 8 cases during their infants period CT scanning at 6,12 months,dilaled ventricle was seen in 6,increased extracerebral space in 8,and periventricalar leukomalacia in 2.Conclusion CT scaning is helpful for evaluating brain injury and predicting prognosis in neonates with HIE.

In recent years constant improvements have been made regarding medical and health laws and reg-ulations in China while at the same time clinical teaching, a closely related field, has been conducted in accordance with old rules and regulations. Such a mechanism of clinical teaching, which finds itself hard to operate under the new medical and health laws and regulations, will restrict or even hinder patient care and clinical teaching. For this rea-son, it is imperative for us to find by legal means solutions to such problems as the status, work extent and legal re-sponsibility of those engaged in clinical teaching and set up a mechanism and mode of legal guarantees for clinical teaching, wherein there are laws to abide by.

Objective To observe the therapeutic effect of Laparescopic Cholecystectomy(LC) on acute calculous cholecystitis.Methods The data from 35 patients receiving laparoscopic cholecystectomy for acute calculous cholecystitis were analyzed.Results Successful rate of operation was 100% in all cases with no complications such as biliarducy injury,intestines damage and hemorrhage taken place,and mortality was 0%.Conclusion LC is safe for treatment of acute calculous cholecystitis.

[Objective]To research the efficient way of isolating and proliferating of induced endothelial cells(ECs) ,measure the number of the ECs and observe the main biological characteristic of ECs m vitro during the passage cultivation.[Method]The marrow mononuclear cells(MNCs) in rabbit marrow were isolated by the gradiem centrifugation and cultivated to isolate and induce ECs. The efficiency of isolating and harvesting ECs was measured by the number of the first passage ECs from every 10~6 of the MNCs. The changes of ECs morphology, growth and proliferation were observed during the passage cultivation. The immunohistochemical staining of CD31 and vWF and function of ECs were checked.[Result]After cultivated at 1?10~6/cm~2 MNCs for 7～8 d, the first passage ECs were harvested. The number of the first and the second passage ECs had a remarkable statistical positive correlation with the number of the MNCs from one rabbit. After cultivated for 7～8 d, the first passage ECs of 7.086?0.75?10~4 in number and the second passage ECs of 53.20?6.47?10~4 in number were harvested from every 10~6 MNCs on average;ECs, which were monolayer arrayed to form cobblelike shape,The cells of 2nd passage showed positive staining of CD31 and vWF, and positive labeling of DiI-LDL and UEA.[Conclusion]The period from cultivating to harvesting ECs is shortened at the density of 1?10~6/cm~2 MNCs. It is feasible that the efficiency of isolating and harvesting ECs could be measured by the number of the first or the second passage ECs from every 10~6 MNCs. The induced ECs can act as seeding cells in bone tissue engineering in future.

Eighty six patients with influenza-like diseases were randomly assigned to trial group (n=42,oseltamivir 75 mg,twice daily for 2 days)and control group(n=44,osehamivir 75 mg,twice daily for 5 days).The results showed that there Was no statistical difierence in the duration of fever between trial groups[(56±14)h]and control group[(54±13)h,P=0.623],the cost of the trial group[(193±38)yuan]Was 189 yuan less than that in the control group[(382±61)yuan,P=0.016].It is suggested that the efficacy of two-day course of oseltamivir is the same as that of five-day course in tlle treatment for influenza-like diseases,but with lower cost.

Objective To investigate the clinical features of and gene mutations in a Chinese Han pedigree with piebaldism. Methods Clinical data were collected with informed consent from a pedigree with piebaldism, processed and documented. A clinical genetic analysis was conducted and pedigree chart was drawn. Genomic DNA was extracted from the peripheral blood of 14 patients and 40 unaffected individuals in the family as well as 50 unrelated human controls, and subjected to the amplification of 21 exons and flanking sequences of the KIT gene by PCR. Sequence analysis was performed by Mutation SurveyorTM. Results There were 73 members in the family, and of them, 14 were diagnosed with piebaldism according to typical clinical features. Piebaldism was inherited in an autosomal dominant pattern in this family. A heterozygous 4-base insertion mutation 1900insATGA in exon 13 of KIT gene was identified in all the 14 affected family members, which resulted in a frame-shift mutation at codon 634 and produced a premature translation termination codon. This mutation was undetected in either the unaffected family members or unrelated controls. Up to the time of this writing, this mutation had not been previously reported. Conclusion The novel mutation 1900insATGA in the KIT gene may be the cause of clinical phenotype of piebaldism in the family.