Objective: The genetic diversity of Paris polyphylla var. yunnanensis germplasmic resources in natural populations was analyzed by simple sequence repeat marker for protection and rational utilization. Methods: A total of 115 individuals of P. polyphylla var. yunnanensis collected from five natural populations were analyzed by SSR marker for genetic diversity analysis. Results: Eight pairs of SSR primers were screened. A relatively high level of genetic diversity was revealed: The percentage of polymorphic bands was 100%, the polymorphism information content was 0.745 6. At the population level and species level, the observed number of allele was 8.425 0 and 17.750 0, respectively, the effective number of allele was 4.960 9 and 7.500 7, respectively, the observed heterozygosity was 0.295 5 and 0.294 8, respectively, the expected heterozygosity was 0.654 8 and 0.774 4, respectively, and the Shannon's information index was 1.520 1 and 2.038 6, respectively. The genetic differentiation coefficient was 0.172 8 and the gene flow was 1.196 6. Based on UPGMA cluster analysis, the five populations of P. polyphylla var. yunnanensis were divided into two clades. Conclusion: The level of genetic diversity of P. polyphylla var. yunnanensis was relatively high and there was certain genetic differentiation within and among populations. This study puts forward several suggestions for the protection and development of P. polyphylla var. yunnanensis resources, which will be helpful for protection and sustainable utilization of scientific advice and reference.

Background Induced pluripotent stem cells (iPSCs)can differentiate into various types of somatic cells without causing ethical controversy and immune rejection in clinical activity,which is similar to differentiation ability of embryonic stem cells.So,iPSCs may be used as seed cells for tissue engineering corneal endothelial reconstruction.Objective The present study was to survey the morphologic change of iPSCs after coculture with corneal endothelium cells(CECs) under the atomic force microscopy(AFM).Methods Rabbit CECs and human MMC-iPSCs were isolated and cultured respectively.The iPSCs were identified with the marker by immunochemistry.iPSCs passaged for 7 days were then cultured with 60％ confluent CECs to establish the co-culture model.The surface morphology and cellular membrane ultrastructure of differentiated iPSCs after induced by CECs were examined by AFM combination with inverted microscope,and compared with CECs and undifferentiated iPSCs.Results Thelengthand width were(66.93±10.48)μm and (44.85 ± 8.14) μm in CECs,(12.51±1.40)μm and (10.93 ±1.69) μm in uninduced iPSCs,and(36.12±10.29) μm and(31.53±9.65)μm in CECs-induced iPSCs.Both the length and width values of CECs-induced iPSCs were statistically bigger than those uninduced iPSCs,with significant differences between them (P＜0.05),but no significant difference was seen in the width valne of CECs-induced iPSCs in comparison with CECs(P＞0.05).The convex structure of CECs cytomembrane surface showed the digitation in shape with the size and height(2.11 ± 1.03) μm and (115.68±92.08) nm respectively,and the concave structure of cytomembrane surface of CECs was fenestrae-like depression and the size was (1.49 ± 0.65) μm.The numerical valuc of mean square root roughness (Rq)and average roughness (Ra)of cytomembrane surface of CECs were(39.20±7.82)nm and (30.37±5.32)nm respectively.The convex surface of cytomembrane of iPSCs was granular-like in shape with size and height(0.39±0.22)μm and(13.11±9.18)nm respectively.The concave surface of cytomembrane of iPSCs was worm-eaten-like concave with the size(0.34±0.18)μm.The numerical value of Rq and Ra of geometrical parameters of cytomembrane surface of iPSCs were (26.60 ± 4.93)nm and (9.97 ± 3.78) nm respectively.The convex surface of cytomembrane of induced iPSCs was digital-like in shape with the size and height (1.91±0.76) μm and(106.55±77.27) nm respectively.The concave surface of cytomembrane of induced iPSCs was fenestrae-like depression and the size of concave was(1.6l±1.25) μm.The numerical value of Rq and Ra on surface of cytomembrane of induced iPSCs was (57.33± 12.80) nm and (43.63± 11.17) nm respectively.The numerical values of the size and height of convex,the size of concave,Rq and Ra on surface of cytomembrane in induced iPSCs were statistically bigger than in iPSCs(P＜0.05)and were not significant differences in comparison with CECs (P＞0.05).Conclusions Morphology of iPSCs translate toward the CECs after induce for 7 days under the AFM.This outcome lays the foundation for further study on iPSCs.

OBJECTIVETo study the value and the clinical application of the Medical Image three-dimensional Visualization System of Abdomen (MI-3DVS) in diagnosis and evaluating resectability of pancreatic tumor.

METHODSTwelve patients with pancreatic tumor were tested with 64-slice helical CT (64-MSCT) angiography, and the CT data was reconstructed with MI-3DVS from November 2008 to August 2009. The 3D findings were adopted in diagnosis and evaluating resectability, and the results were compared with surgical operation and the pathological finding. There were 7 male and 5 female, aged from 14 to 83 years. Within the 12 cases, there were 4 cases with pancreatic carcinoma, 5 cases with pancreatic solid pseudopapillary tumor, 2 cases with pancreatic serous cystadenoma, 1 case with pancreatic cyst (ductal epithelial papillary hyperplasia).

RESULTSNine tumors which had been regarded as removable pre-operatively with MI-3DVS were removed successfully. Three patients who were considered unresectable by other hospitals with CT were operated successfully with MI-3DVS. The other 3 patients' tumors were actually not able to be removed as pre-operative evaluation.

CONCLUSIONMI-3DVS plays an important role in diagnosis and assessment of resectability of pancreatic tumor.

Adolescent ; Adult ; Aged ; Female ; Humans ; Imaging, Three-Dimensional ; Male ; Middle Aged ; Pancreatic Neoplasms ; diagnostic imaging ; surgery ; Radiography, Abdominal ; methods ; Tomography, Spiral Computed ; Young Adult

In 2012, the preparation process and quality standard for Guizhi Fuling capsule were improved. To compare the effects and differences of capsules before (2011) and after(2012-2014) the improvement, evaluation models for intrinsic dysmenorrhea, pelvic inflammation and hysteromyoma were applied in rats. Models were induced by oxytocin, liqiud bacteria mixture and estrogen loading, respectively. The capsules (12 batchs/year, 48 bathcs in all), sampled randomly in 2011-2014, the effects were assessed using the three models. In anti-dysmenorrhea models, remarked reduction of writhing frequency, ET-1 and PGF2α content in uterus could be detected, as well as extension of writhing latency. In pelvic inflammation rats, depression of TNF-α and raise of IL-2 were induced by earh batch of capsules. In hysteromyoma model, uterine weight and smooth muscle proliferation, including E2 and P level in plasma, were lowered obviously by all batchs of capsules. Secondly, Guizhi Fuling capsules produced in 2012-2014 revealed better effectiveness than the ones manufactured in 2011. Moreover, pharmacodynamics indexes of the samples made in 2011 differed significantly between groups, which could not be observed in the ones ot 2012-2014. After tne preparation process and quality standard improvement, the effectiveness and homogeneity of Guizhi Fuling capsules were enhanced.
Animals ; Capsules ; administration & dosage ; chemistry ; standards ; Depression ; drug therapy ; genetics ; metabolism ; Dinoprost ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; standards ; Dysmenorrhea ; drug therapy ; genetics ; metabolism ; Female ; Humans ; Interleukin-2 ; genetics ; metabolism ; Pelvic Inflammatory Disease ; drug therapy ; genetics ; metabolism ; Quality Improvement ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

OBJECTIVETo investigate the role of the key intracellular signaling molecule glycogen synthase kinase-3 beta in the mechanism of liver ischemia reperfusion (IR).

METHODSC57BL/6 mice were subjected to 90 min warm liver cephalad lobe ischemia, followed by various length of reperfusion. Experiment groups included sham control group, liver IRI model group and glycogen synthase kinase-3 beta inhibitor-treated group (SB216763 in DMSO, 25 g/kg, i.p, 2 hour prior to the onset of liver ischemia). The expression of glycogen synthase kinase-3 beta protein was analysed by Western blotting. The serum ALT levels were determined to reflect the function of liver. The affected liver lobes were harvested for histology analysis. The inflammatory gene expression was detected by Quantitative PCR.

RESULTSBy western blot analysis, we found that ischemia itself activated glycogen synthase kinase-3 beta by a significant decrease of its phosphorylation. Glycogen synthase kinase-3 beta inhibitor SB216763-pretreatment ameliorated the liver damages significantly as compared to the controls (sALT: 2046+/-513 U/L vs 5809+/-1689 U/L, P = 0.0153), and suppressed the gene expressions of IL-12, TNFa, IL-1b and IL-6.

CONCLUSIONSThis study demonstrated that the ischemia process modulated liver innate immune activation via a GSK-3-dependent mechanism which favored the development of a pro-inflammation response and lead to liver tissue damages. GSK-3b may be a new therapeutic target to ameliorate liver IRI in transplant patients.

Animals ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Inflammation ; metabolism ; Liver ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Reperfusion Injury ; metabolism ; pathology

Objective To study midterm curative effect of domestic drug eluting coronary stent in treatment of left main (LM) lesion.Method From April 2004 to September 2005,totally 32 patients with LM lesion received percutaneous coronary intervention (PCI) without protection.Thirty-nine coronary stents (Firebird, Microport,Shanghai) were implanted.One stent was used when the lesion only involved the opening or the middle portion of the LM branch.If the bifurcation was involved,the stenosis was treated considering the following factors: the diameter of left main,left anterior descending (LAD) and the left circumflex branch (LCX);whether there was stenosis of the opening of circumflex branch.Methods included"T"stent,stent pass though the opening of circumflex branch and kissing stents.All patients were followed at 1,6 and 12 months after discharge.Coronary angiography of patients was performd at 5 months after discharge.Central necrosis,restenosis,target vessel reconstructive rate (TVR) and target lesion reconstructive rate (TLR) were analyzed.Results Most LM lesion involved bifurcation (53.1%).Treatment for bifurcation included"T"stent in 47% patients,stent passing through the opening of LCX in 35.3 %.No restenosis occurred in lesion involving the opening or middle portion of the LM during follow-up.The 62.3% patients received coronary angiography.Angiography showed 5% of restenosis rate,5% TLR,10% TVR and 6.3% central necrosis rate.Conclusions Drug eluting stent implantation in low risk unprotective left main lesion may achieve satisfactory clinical results.

OBJECTIVETo explore if folic acid/polyamide-amine (FA/PAMAM) enhances the therapeutic effect of miR-7gene therapy for glioma in vivo.

METHODSThe miR-7 gene was transfected into U251 glioma cells by FA/PAMAM. The efficiency of gene transfection was assessed by fluorescence microscopy. The miR-7 level was detect by quantitative RT-PCR. Intracranial glioma models were established in thymectomized mice, and FA/PAMAM nanoparticles were transplanted into the tumors in situ 3 days later. The animal survival was recorded and the gross tumor volume and degree of edema were observed by MRI. Apoptosis in the glioma cells and expression of proliferating cell nuclear antigen (PCNA), matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9) were assessed by immunohistochemistry, and EGFR and AKT-2 protein expression was detected by Western blot assay.

RESULTSCompared with the liposomes, the FA/PAMAM nanoparticles were more efficient to transfer miR-7 gene into U251 glioma cells, MRI showed that the tumor growth was much slower in the FA/PAMAM/miR-7 group, and the animal survival time was longer. The apoptosis rate was (5.3 ± 0.9)% in the control group, (11.4 ± 2.4)% in the liposome/miR-7 group, and (17.7 ± 3.7)% in the FA/PAMAM/miR-7 group. The immunohistochemical assay showed that the levels of PCNA, MMP-2 and MMP-9 protein in the FA/PAMAM/miR-7 group were (34.6 ± 5.4)%, (24.5 ± 4.1)%, (25.4 ± 5.1)%, respectively, significantly lower than those in the liposome/miR-7 group (49.3 ± 5.9)%, (31.7 ± 7.1)% and (39.4 ± 6.4)%, respectively, and those in the control group (57.3 ± 7.4)%, (45.4 ± 6.9)% and (55.1 ± 7.3)%, respectively (all P < 0.05). The expressions of EGFR and AKT-2 proteins were 1.09 ± 0.12 and 0.62 ± 0.10 in the control group, 0.63 ± 0.11 and 0.43 ± 0.07 in the liposome/miR-7 group, and significantly deceased (0.47 ± 0.09 and 0.31 ± 0.04, respectively) in the FA/PAMAM/miR-7 group (all P < 0.05).

CONCLUSIONCompared with the liposomes, FA/PAMAM can transfect miR-7 into glioma cells with a higher efficiency in vivo, makes a longer time of the drug action, and shows a certain inhibitory effect on the growth of glioma, therefore, might become a new drug targeting agent in gene therapy forglioma.

Animals ; Apoptosis ; Brain Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Dendrimers ; chemistry ; Folic Acid ; chemistry ; Genetic Therapy ; methods ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Nude ; MicroRNAs ; genetics ; metabolism ; Nanoparticles ; Neoplasm Transplantation ; Proliferating Cell Nuclear Antigen ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Receptor, Epidermal Growth Factor ; metabolism ; Thymectomy ; Transfection

OBJECTIVETo observe the changes and characteristics of interdigestive migrating motor complex (MMC) in rat models of acute liver failure.

METHODS30 rat models with acute liver failure were induced with D-galactosamine and another 30 normal rats were used as controls. The indexes of MMC recorded by multi-channel physiological recorder were compared.

RESULTSNo significant differences found between the two groups in antral and duodenal MMC cycles and frequencies of duodenal and jejunal MMC III phase. Compared with normal controls, the MMC II phase in the acute liver failure rats was significantly prolonged (t=-3.97, -3.85, P<0.05), the MMC III duration of antrum and duodenum (u=-4.99, t=4.66, P<0.05) was shorter and the MMC III frequency of antrum (u=-4.73, P<0.05) was faster. In addition, the MMC cycle and MMC III phase of jejunum were significantly prolonged (u=-1.63, t=-4.94, P<0.05) and the MMC III phase duration was significantly shorter in the acute liver failure rats (t=5.10, P<0.05).

CONCLUSIONSignificantly prolonged MMC II phase characterized by migrating clustered contraction, shortened MMC III phase and extended jejunal MMC cycles were probably the major contributors to the gastrointestinal motility disorders in the rats with acute liver failure.

Animals ; Liver Failure, Acute ; physiopathology ; Myoelectric Complex, Migrating ; Rats ; Rats, Wistar

This study was aimed to investigate the expression of blood Th17 and CD4(+) CD25(+) regulatory T cells (Treg) in the patients with aplastic anemia (AA). Forty-five patients with AA were enrolled into this study, and were divided into mild aplastic anemia (MAA) group (n = 25) and severe aplastic anemia group (SAA) (n = 20), blood cell count was recorded. 15 healthy donors were enrolled as control. Proportions of blood Th17 and CD4(+) CD25(+) Treg cells were determined by flow cytometry. The serum levels of IL-17, IFN-γ and TNF-α, as well as their concentrations in culture supernatant of phytohemagglutinin (PHA) -stimulated peripheral blood mononuclear cells, were measured by ELISA. The results showed that the proportions of blood Th17 cells and concentration of blood serum IL-17 and IFN-γ increased in patients with SAA, compared with MAA and normal controls, but CD4(+) CD25(+) Foxp3(+) Treg cells obviously decreased in patients with SAA. The concentrations of IL-17 and IFN-γ significantly increased in culture supernatant of SAA group. Hemoglobin level in the patients with AA negatively correlated with the population of Th17 cells and serum IL-17 level, whereas positively correlated with the expression of CD4(+) CD25(+)Treg cells. It is concluded that the increased response of Th17 cells and deficiency of CD4(+) CD25(+) Treg cells present in severe aplastic anemia. The severity of anemia may be related with the imbalance between Th17 and CD4(+) CD25(+)Treg cell response.
Adult ; Aged ; Anemia, Aplastic ; blood ; metabolism ; Case-Control Studies ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-17 ; blood ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; metabolism ; Th17 Cells ; metabolism ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

OBJECTIVETo study the applied value of abdominal medical image proceeding system (AMIPS) in extended hepatectomy.

METHODSSixty-four-slice spiral computer tomography (CT) scan data of 32 cases treated from September 2007 to July 2009 (15 male, 17 female; mean age 52 years old) with liver tumor was collected, among which there were 9 cases with huge liver tumor. The data was imported into AMIPS for sequence segmenting and three-dimensional (3D) reconstruction. The reconstructed models were imported into virtual system of AMIPS for digital hepatic segment partition and extended hepatectomy analysis for huge liver tumor of 9 cases. According to the calculated data, suitable modus operandi were selected.

RESULTSIn the AMIPS, the 3D models could show the relationships between the lesions to the surrounding tissue more intuitively and the type of blood supply. Digital hepatic segment partition made localize lesions more exactly. It was possible to hepatic segmentectomy and analysis of extended hepatectomy by calculating the relative volume of hepatic segment of huge liver tumor. Huge liver tumor of 9 cases performed different modus operandi, including two cases with right hemi hepatectomies, five cases with hepatic segmentectomy of S6 and S7, two cases with hepatic segmentectomy of S5-7 and part of the S8. There were margin-free tumor cells and no complications such as liver failure in all cases. The average hospitalization time was 21 d.

CONCLUSIONAMIPS is helpful in the diagnosis of hepatic disease and in the optimizing surgical plans which can decrease surgical risk and help prevent postoperative hepatic failure.

Computer Simulation ; Female ; Hepatectomy ; methods ; Humans ; Imaging, Three-Dimensional ; Liver Neoplasms ; surgery ; Male ; Middle Aged ; Tomography, Spiral Computed ; User-Computer Interface