AIM:To observe the sustained-release effect of compound betamethasone by subconjunctival injection on immunological rejection after ostrich-rabbit lamellar keratoplasty.METHODS:Sixteen healthy New Zealand white rabbits with 6wk old received corneal lamellar keratoplasty,and the corneal graft was ostrich acellular corneal stroma.After surgery all subjects were divided into two groups,Group A (experimental group) were administrated with subconjunctival injection of compound betamethasone injection (once every 7d),and Group B (control group) were administrated with subconjunctival injection of dexamethasone sodium phosphate (once every 7d).At 1,2wk,1,2mo after the surgery,rabbit corneas were taken for paraffin sections,and were observed with H-E staining,in the meantime changes of CD4+ and CD8+ T lymphocytes were observed by immunofluorescence.RESULTS:Two months after surgery,in Group A corneal grafts remained transparenct,and showed little neovascularization;HE staining and indirect immunofluorescence showed that only a few neutrophil infiltration,no CD4+ and CD8+T lymphocytes.In Group B,the inflammatory reaction was observable at different time points,the corneal graft was turbid;and the tissue sections and indirect immunofluorescence staining showed that neutrophil infiltration was predominant,and CD4+,CD8+T lymphocytes were also seen.CONCLUSION:Compound betamethasone is able to inhibit the ostrich-rabbit corneal transplantation immune rejection,prolong the survival time of the grafts.The present study lay the foundation for further research and clinical application.

AIM: To study the roles of tumor necrosis factor alpha ( TNF-α) , interferon gamma ( IFN-γ) in pathogenesis of primary pterygium and to explore the mechanism of tears in pterygium.?METHODS: Concentrations of TNF-α and interferon gamma IFN - γ were quantified by enzyme - linked immunosorbent assay ( ELISA ) in asymptomatic control group (30 eyes of 30 healthy volunteers) and 60 primary pterygium patients ( 30 eyes of the 30 progressive cases, 30 eyes of the 30 quiescent cases) . The data was analyzed by analysis of variance.?RESULTS:The concentration of TNF-αand IFN-γin the patients with primary pterygium were statistically significant (P<0. 05), compared with the normal control group. Tear concentration of TNF-α was significantly higher in primary pterygium compared with the control group. Tear concentration of IFN - γ was significantly reduced in primary pterygium compared with the control group (P<0. 05).?CONCLUSION:TNF-α and IFN-γmay be important in the pathogenesis and development of pterygium tissues.It can provide some new directions and ideas for the treatment of primary pterygium.

OBJECTIVETo explore the effect of Caspase 1 inhibitor Ac-YVAD-CMK on acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation(allo-HSCT) and its mechanism.

METHODSExperiments were divided randomly into 3 groups: allogeneic hematopoietic stem cell transplantation combined with splenic cell infusion group (TS group, n=12), allogeneic hematopoietic stem cell transplantation combined with splenic cell infusion plus injection of low dose Caspase 1 inhibitor group (TS+low dose of C group, n=16) and plus high dose Caspase1 inhibitor (TS+high dose of C group, n=19). The body weight of mice in each group was dynamically detected, and the clinical manifestation of GVHD and score of aGVHD were determined, and the chimerism rate of mice was detected after transplantation for 14 days. Th1, Th2 and Th17 cells in peripheral blood were examined by flow cytometry. Peripheral proinflammatory cytokines IL-1β, IFN-γ, IL-1α and IL-18 were examined by enzyme-linked immunosorbent assay(ELISA). The tissues sections of GVHD target organs (liver, lung, colon and skin) were stained with HE for histopathologic examination and histopathologic score.

RESULTSAc-YVAD-CMK could alleviate murine aGVHD and pathological injury, decreare the incidence and severity of aGVHD in recipient mice. The detection of Th cell subsets in peripheral blood by flow cytometry showed that compared with TS group, the Th1 cell ratio in TS+low dose of C and TS+high dose of C groups was significantly reduced (P<0.05), while the Th2 and Th17 cell ratio was significantly enhanced (P<0.05) in TS+low dose of high dose of C groups. The detection of peripheral inflamematory cytokines by ELISA demonstrated that the inflammatory cytokines including IL-1β,IFN-γ,IL-18 and IL-1α were reduced significantly (P<0.05).

CONCLUSIONAc-YVAD-CMK can improve aGVHD by inhibiting Caspase 1 and reducing the release of some inflammatory cytokines, thereby alleviated the aGVHD pathological damage.

Objective To optimize the extraction conditions of Schisandra sphenanthera(SS)seed oil by supercritical CO2ex?traction and identify its components by GC-MS.Methods SS seed oil was used as tested material,response surface methodology was used to optimize the process of supercritical CO2extraction,and GS-MS method was used to analyse SS seed oil composition.Results A model of an equation was established,which could be used to optimize the process parameters of supercritical CO2extraction of SS seed oil. The optimum extraction parameters were as follows:the extraction pressure was 33 MPa,the extraction temperature was 53℃,the extraction time was 90 min,the CO2flow rate was 21.40 ml/min.In this condition,the extraction rate of SS seed oil was 7.97%.The SS seed oil was analyzed by GC-MS,and 23 compounds were identified.In these compounds,(1α,4a.β,8a.α)-1,2,3, 4,4a,5,6,8a-octahydro-7-methyl-4-methylene-1-(1-methylethenyl)-naphthalene,(-)-1,7-dimethyl-7-(4-methyl-3-pentenyl)-tricyc?lo[2.2.1.0(2,6)]heptane and(R)-2,4a,5,6,7,8-octahydro-3,5,5,9-tetramethyl-1H-(σ-phenyl)cycloheptene had the content of more than 10% and the contents were 27.78%,14.77% and 13.12% respectively.Conclusion This process has high extraction rate, fast speed and simple operation,and can be used for the extraction of SS seed oil.

To investigate the overall intestinal permeability of multiple components in lotus leaves and make clear the interaction in composition absorption process. Rat single-pass intestinal perfusion technique was used, and the results showed that the Peff values of nuciferine, demethylanuciferine, rutin, quercetin, kaempferol from lotus leaf were greater than 0.5×10⁻⁴ cm•s⁻¹. In the biopharmaceutics classification system (BCS) intestinal permeability property, these ingredients were high permeable components, while the hyperin was low permeable component. However, in the multi-component environment of the lotus leaf extract, component permeation was changed. Semi quantitative analysis of the unclear components showed that under the multi-component environment, four in seven components with relatively high contents had a Peff value less than 0.5×10⁻⁴ cm•s⁻¹, indicating these 4 components were of low permeability, while other 3 components were of high permeability. The results could be valuable to make clear the overall intestinal permeability of multiple components in lotus leaf, and lay a foundation for studying the mechanism of the lipid-lowering effect of lotus leaf.

The objective of the present study was to establish a method based on principal component analysis (PCA) for the study of transdermal delivery of Chinese medicinal formulae, and to choose the best penetration enhancers for Qingfei Xiaocuo gel depend on this method. Using improved Franz type diffusion cell and excised rat skin in vitro as transdermal barrier, the receptive solution fingerprint was established by HPLC, harvesting the areas of the common peaks in the fingerprint, then the total factor scores of the concentrations at different times were calculated using PCA and were employed instead of the concentrations to compute the cumulative amounts (Q12) and enhancement ratio (ER), the latter of which were considered as the indexes for optimizing penetration enhancers. Compare to the control group, the ER of the other groups increased significantly and furthermore, 2.5% azone with 2.5% menthol manifested the best effect. PCA represent most information in the receptive solution, the method above could choose the best penetration enhancers, it could be a reference for the study of transdermal delivery of Chinese medicinal formulae.
Administration, Cutaneous ; Animals ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Gels ; In Vitro Techniques ; Male ; Medicine, Chinese Traditional ; Mice ; Principal Component Analysis ; Skin ; metabolism

OBJECTIVETo evaluate the effects of two gastric cancer screening schemes for early detection of gastric cancer in a high-risk population.

METHODSA cluster random sampling method was used to select local residents aged 40-69 years from Linqu County, Shandong Province. "Serum pepsinogen initial screening combined with further endoscopic examination (PG scheme)" and "direct endoscopic examination (endoscopy scheme)" were conducted. The associations between screening schemes and detection rates of gastric cancer, and early gastric cancer/high-grade intraepithelial neoplasia were evaluated by unconditional logistic regression analysis.

RESULTSOverall, 3654 and 2290 participants completed PG and endoscopy schemes, respectively. A total of 11 (0.30%) cases of gastric cancer and 10 (0.27%) cases of high-grade intraepithelial neoplasia were detected by PG scheme, of which 7 (0.19%) cases were early gastric cancer. While, 19 (0.83%) cases of gastric cancer and 10 (0.44%) cases of high-grade intraepithelial neoplasia were detected by endoscopy scheme, with 12 (0.52%) cases of early gastric cancer. Compared with the PG scheme, the endoscopy scheme had a significantly higher detection rates of gastric cancer (OR = 2.83, 95%CI 1.34-5.98), and early gastric cancer/high-grade intraepithelial neoplasia (OR = 2.12, 95%CI 1.12-4.02).

CONCLUSIONSThe endoscopy scheme is more effective in the detection of gastric cancer in a high-risk population, particularly for early gastric cancer/high-grade intraepithelial neoplasia than the PG scheme.

Adult ; Aged ; Carcinoma ; blood ; diagnosis ; Carcinoma in Situ ; blood ; diagnosis ; Early Detection of Cancer ; methods ; Female ; Gastroscopy ; Humans ; Male ; Mass Screening ; methods ; Middle Aged ; Pepsinogen A ; blood ; Stomach Neoplasms ; blood ; diagnosis

This study was aimed to investigate the effect of Wnt/β-catenin signaling pathway on the biologic behavior of mouse bone marrow mesenchymal stem cells (mBM-MSC) by constructing a RNAi lentiviral vector specific to β-catenin. Three pairs of shRNA coding sequences directed against different sites of β-catenin mRNA were designed and were linked into lentiviral vector plasmid PLB for constructing the PLB-β-catenin/shRNA1, PLB-β-catenin/shRNA2 and PLB-β-catenin/ shRNA3. Those plasmids and lentiviral packaging plasmids were co-transfected into the packaging cells 293FT, then the virus particles were collected and the viral titer was assayed after concentration, and these viral particles were infected to MSC. The flow cytometry was used to sort GFP (+) cells, Western blot and RT-PCR were used to verify the inhibitory effect of those cells on expression of β-catenin gene in cells, CCK-8 method was used to detect the cell proliferation level, Annexin-V/7-AAD was used to determine the cell apoptosis after interference, the cell scratch and transwell tests were used to detect the migration capability of MSC. The results showed that the efficient inhibition of β-catenin mRNA and protein expression, and the suppression of MSC proliferation were observed in group of PLB-β-catenin/shRNA2 (interference group), while there was no significant changes of MSC proliferation between negative group (PLB group) and the normal group (control group). The flow cytometric detection indicated that after induced by serum starvation for 72 h, the apoptosis of MSC increased in interference group, but there was no difference between PLB and control groups (P > 0.05). The cell scratch and transwell tests demonstrated that the migration capability of MSC in interference group decreased significantly, while the migration capability of MSC in control group was not changed obviously. It is concluded that the constructed specific RNAi lentivirus can effectively inhibit the expression of β-catenin gene, decrease expression level of β-catenin mRNA and protein. The Wnt/β-catenin signaling pathway plays an important role in biological behavior of BM-MSC.
Animals ; Apoptosis ; Bone Marrow Cells ; cytology ; Cell Line, Tumor ; Cell Movement ; Genetic Vectors ; Lentivirus ; genetics ; Mesenchymal Stromal Cells ; cytology ; Mice ; RNA Interference ; RNA, Small Interfering ; Wnt Signaling Pathway ; beta Catenin ; metabolism

BACKGROUNDIntraoperative magnetic resonance imaging (iMRI) dates back to the 1990s and has been successfully applied in neurosurgery but they were low-field iMRI (< 1.0T). This paper reports the clinical experience with a 3T iMRI-integrated neurosurgical suite in Huashan Hospital, Shanghai, China.

METHODSFrom September 2010 through March 2012, 373 consecutive patients underwent neurological surgery under guidance with 3T iMRI. A retrospective analysis was conducted regarding clinical efficiency.

RESULTSAll surgery in the 373 patients was safe. The ratio of gross total resection for cerebral gliomas (n = 161) was increased from 55.90% to 87.58%. The ratio of benefit in extent of resection was 39.13%. One hundred and fifty eight of the 161 glioma patients accomplished follow-up at 3 months postoperatively. Twenty of 161 patients (12.42%) suffered from early motor deficit after surgery. Late motor deficit was however observed in five of 158 patients (3.16%). Twenty-one of 161 patients (13.04%) had early speech deficit and late speech deficit was only observed in six of 158 patients (3.80%). The ratio of gross total resection for pituitary adenomas (n = 49) was increased from 77.55% to 85.71%. The ratio of benefit in extent of resection was 10.2%. There were no iMRI-related adverse events even for patients who underwent awake craniotomy.

CONCLUSIONThe 3T iMRI integrated neurosurgical suite provides high-quality intraoperative structural and functional imaging for real-time tumor resection control and accurate functional preservation, resulting in an improvement in maximal safe brain surgery.

Adolescent ; Adult ; Aged ; Brain Neoplasms ; surgery ; Child ; Child, Preschool ; China ; Female ; Glioma ; surgery ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Neurosurgical Procedures ; methods ; Young Adult

OBJECTIVETo analyze the meiotic segregation results of the spermatozoa from male pericentric inversion carriers by fluorescence in-situ hybridization (FISH).

METHODSUsing chemical depolymerization and multicolor FISH, we analyzed the meiotic segregation results of the spermatozoa from 4 male pericentric inversion carriers.

RESULTSOf the 4 males studied, 46,XY,inv(9) (p11q12) was found in 2, 46,XY,inv(9) (p11q13) in 1 and 46,XY,inv(6) (p22q24) in the other; the lengths of the inverted segments represented 16.0, 16.0, 21.0 and 76.0% of the size of the whole chromosome involved; and the frequencies of recombinant sperm were 0.2, 0.4, 0.3 and 43.9%, del(p)/dup(q) accounting for 22.4% and del(q)/dup(p) 21.5%, respectively.

CONCLUSIONMales with pericentric inversion may produce spermatozoa with recombinant chromosomes and the rate of recombination varies principally according to the size proportion to the whole chromosome involved. The results of FISH analysis of chromosomal unbalanced spermatozoa can provide accurate personalized information on the genetic risk of fertility.

Adult ; Chromosome Inversion ; genetics ; Chromosomes, Human, Pair 9 ; genetics ; Heterozygote ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infertility, Male ; genetics ; Male ; Meiosis ; Spermatozoa