Animals ; Cells, Cultured ; Interleukin-1beta ; pharmacology ; Male ; Neurons ; drug effects ; metabolism ; Nitric Oxide ; metabolism ; Nitriles ; toxicity ; Pyrethrins ; toxicity ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase ; metabolism

Objective To develop an immunomagnetic bead ( IMB) assay for quantitative detection enolase ( Eno) of Candi-da albicans, and to improve the diagonosis of invasive candidiasis . Methods The immunomagnetic bead was prepared by conjuga-ting with Anti Eno of Candida albicans monoclonal antibody .HRP-conjugated goat polyclonal antibody against Candida albicans Eno was employed as detecting antibody .The performance parameters of the IMB assay including precision , specificity, linear range and limit of detection were verified by using recombinant Candida albicans Eno.Then the developed assay was applied to determine Eno levels in supernatant of pathogenic fungi cultures . Results The intra and inter-coefficient of variation was 4.54%, 5.87% and 5.26%, 8.82%at the concentration of 25 ng/mL and 5 ng/mL, respectively.The limit of detection was 0.5 ng/mL.The linear range was(0.5-50) ng/mL.The level of Eno in Candida albicans culture after incubated in 37℃for 24 h was 3.89 ng/mL and gradually in-creased to 37.89 ng/mL at 120 h.There was a positive correlation between the level of Eno and growth hyphae of Candiad albicans. There was weak cross reaction with Candida parapsilosis and no cross reaction with Candida tropicalis, Candida guilliermondii, Candida glabrata, Cryptococcus neoformans and Saccharomyces cerevisiae. Conclusion An IMB assay for quantitative detection Eno of Candida albicans was developed , which was more sensitive , rapid and reliable than previous qualitative ELISA .The IMB assay has the potential to be applied to the research in invasive candidasis .

Adenocarcinoma ; pathology ; surgery ; Aged ; Carcinoma, Signet Ring Cell ; pathology ; surgery ; Cell Differentiation ; Female ; Follow-Up Studies ; Gastrectomy ; methods ; Humans ; Male ; Middle Aged ; Paneth Cells ; pathology ; Stomach Neoplasms ; pathology ; surgery

NOD mice were treated intraperitoneally with tripterygium wilfordii ployglycosidium (TWP) for 4 weeks to observe the incidence of cyclophosphamide accelerated diabetes.The apoptosis of?-cells was detected by TUNEL,the expression of caspase-3 in islets of the NOD mice was detected by immunohistochemistry and the expression of caspase-8 mRNA in pancreas by RT-PCR.The results revealed that the incidence of diabetes in TWP group was lower than that in control group.The apoptosis index of?-cells was decreased in TWP group. The expression of caspase-3 in islets and the expression of caspase-8 mRNA in pancreas in TWP group were lower than those in control group.

Aged ; Female ; Hodgkin Disease ; diagnosis ; pathology ; Humans ; Reed-Sternberg Cells ; pathology

OBJECTIVETo optimize the method in the Chinese Pharmacopoeia for determining Salviae Miltiorrhizae Radix et Rhizoma.

METHODTanshinone II(A) and salvianolic acid B were selected as the index in optimization of the sample preparation method of Salviae Miltiorrhizae Radix et Rhizoma in Chinese Pharmacopoeia. Orthogonal test was used to optimize the extraction process of Salviae Miltiorrhizae Radix et Rhizoma, and concentration of contents were detected by high performance liquid chromatography method. A detection of using methanol-water (85: 15) at wavelength of 270 nm was employed for tanshinone II(A) and a detection of using methanol-acetonitrile-formic acid-water (30:10:1: 59) at wavelength of 286 nm was employed for salvianolic acid B.

RESULTThe optimized extraction process of tanshinone II(A) and salvianolic acid B was: extracted by 90% methanol and reflux twice (0.5 h each time) at 75 degrees C, extracted by 70% methanol and reflux twice (1.5 h each time) at 75 degrees C, respectively.

CONCLUSIONOptimized extraction and determination methods could be used to reflect the content of tanshinone II(A) and salvianolic acid B in Salviae Miltiorrhizae Radix et Rhizoma more accurately and efficiently.

Benzofurans ; analysis ; Chromatography, High Pressure Liquid ; Diterpenes, Abietane ; analysis ; Rhizome ; chemistry ; Salvia miltiorrhiza ; chemistry ; Temperature

Adenocarcinoma, Mucinous ; pathology ; Aged ; Appendectomy ; methods ; Appendiceal Neoplasms ; pathology ; surgery ; Appendicitis ; pathology ; Appendix ; pathology ; Carcinoid Tumor ; pathology ; surgery ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Male

Objective To explore the genetic defects in patients with pulmonary atresia (PA).Methods Twenty-three patients with PA were studied from recent 1 year.There were 15 boys and 8 girls,aged 2 days to 10 years.Among them,there were 3 cases with complete ventricle septum,20 cases with VSD,3 cases without main pulmonary trunk,20 cases with PDA,6 cases with integrated major aorto-pulmonary collateral arteries(MAPCAs),3 cases with integrated right aortic arch,2 cases with integrated extra-cardiac abnormality with the manifestation of facial abnormality,1 case with integrated aberrant right subclavian artery,and 1 case with integrated tracheal bronchus.The chromosomes of the children were routinely analyzed.Negative chromosomes were examined to identify the 22ql 1.2 microdeletion by fluorescence in situ hybridization (FISH) test.Results One case of trisomy syndrome was found.22ql 1.2 microdeletion was found in 3 cases.Among 22q1 1 microdeletion cases,1 case was intact ventricular septum,2 cases were integrated VSD.22q1 1.2 microdeletion was not found in the rest 19 cases.Conclusions Twenty-one trisomy syndrome and Del 22q1 1.2 syndrome may play an important role in the pathogenesis of PA.TBX1 probe can be used to examine 22q1 1.2 microdeletion.Del 22q1 1.2 syndrome was suspected in cases of incorporating aberrant subclavian artery,MAPCAs and severe poor development of pulmonary artery.

Autopsy ; Eye Injuries ; Forensic Medicine ; Humans

OBJECTIVECord blood (CB) provides a rich source of stem cells for transplantation. CB transplantation has been used widely after myeloablative therapy. One major disadvantage of CB transplantation is delayed platelet engraftment. The aim of this study was to hasten platelet engraftment by investigating the ability of different hematopoietic growth factor combinations to generate large numbers of megakaryocyte (Mk) from CB and by evaluating the biologic characteristics and function of the expanded Mk.

METHODSCB samples were obtained at the end of normal full-term deliveries with informed consent. Mononuclear cells (MNCs) were isolated from CB using Ficoll density centrifugation. MNC population was positively selected for CD(34) expression by magnetic cell sorting (MACS). CD(34)(+) cells were cultured in serum-free and stroma-free medium containing the following two different cytokine combinations: thrombopoietin (TPO) + stem cell factor (SCF) + interleukin (IL) -3 + IL-6 and TPO + SCF. Cultures were characterized after 3, 7, 10 and 14 days by flow cytometry, colony forming unit-megakaryocyte (CFU-Mk) and maturation evaluation (Mk ploidy). The expanded Mk function was examined by the platelet activation in vitro and severe combined immunodiffiency (SCID) mice transplantation in vivo.

RESULTSDifferent results were observed with different culture conditions. With the first cytokine combination optimal expansion of CD(41)(+) cells was observed on day 10, but the optimal expansion of Mk progenitors (CD(34)(+)CD(41)(+)) was observed on day 7, with a median 121 and 44-fold increase at the starting cell dose. This result was also proven by CFU-Mk. The largest numbers of CFU-Mk were also observed on day 7. The degree of maturation of Mk cells also increased as suggested by DNA content of CD(41)(+) cells, which means that CD(34)(+) cells cultured for 3 - 7 days were richer in primitive Mks, while those cultured for 10 - 14 days had greater numbers of more differentiated Mks. For the second cytokine combination, CD(41)(+) and CD(34)(+)CD(41)(+) cells were fewer than the first one, but it produced 36 and 85-fold CD(34)(+)CD(41)(+) and CD(41)(+) respectively on day 7. Platelet activation test confirmed that the expanded Mks had normal function. Therefore, the expanded Mks could be transplanted into the SCID mice bone marrow and produce human platelet in the peripheral blood of the mice.

CONCLUSIONEx vivo expanded Mk might facilitate CB transplantation and help shorten the period of post-transplant thrombocytopenia.

Animals ; Antigens, CD34 ; Cell Culture Techniques ; methods ; Cells, Cultured ; Culture Media ; Fetal Blood ; cytology ; Humans ; Leukocytes, Mononuclear ; cytology ; Megakaryocytes ; cytology ; Mice ; Mice, SCID