Objective To study the immunogenicity of human embryonic stem cells(hESCs)and the derived neural stem cells(NSCs)in vitro.Methods The constitutive expression of human leucocyte antigen(HLA)Ⅰ and Ⅱ in hESCs and the NSCs derived from these hESCs were detected by flow cytometry (FCM), as well as the expression of HLA-Ⅰ,Ⅱin NSCs induced by 30 ng/ml recombination human interferon-?(IFN-?).Meanwhile, the NSCs before and after induction of IFN-? were co-cultured with peripheral blood lymphocyte obtained from healthy person.Lymphocyte proliferation standing for the immunoreactivity of NSCs was then investigated.Results The hESCs slightly expressed HLA-Ⅰ(6.18%) and hardly any HLA-Ⅱ before differentiation.However, the NSCs expressed more HLA-Ⅰ(23.56%)as well as HLA-Ⅱ(1.28%, 1.73%)than the hESCs did.Both HLA-Ⅰ(46.43%)and HLA-Ⅱ(8.73%, 10.57%)expressed by the NSCs after they were induced by IFN-? were up-regulated.Conclusions hESCs express certain level of HLA-Ⅰ molecules but do not constitutively express HLA-Ⅱ molecules.The derived NSCs express heavy HLA-Ⅰ and a little HLA-Ⅱ, when treated by IFN-? they can inducibly up- regulated both molecules.The NSCs derived from HESCs are of immunogenicity, which induce rejection aiming at HLA-Ⅰ molecules or even at HLA-Ⅱ molecules when the host is inflammative or under stress, which can result in a failure of cellular transplantation.

OBJECTIVETo explore Chinese medical theory of Fei and Dachang being interior-exteriorly correlated by observing changes of inherent immune response and acquired immune response in the lung tissue and the intestinal tissue of ulcerative colitis (UC) model rats and the intervention of Chinese compounds (CM).

METHODSSeventy rats were randomly divided into 5 groups, i.e., the normal control group (n = 10), the model group (n = 15), the treatment 1 group (n = 15, treated from Fei), the treatment 2 group (n = 15, treated from the intestine), and the Western medicine (WM) group [n = 15, treated with Sulfasalazine (SASP). Except those in the normal control group, the UC rat model was prepared by allergizing colon mucosa combined with TNBS-alcohol (50%) enema, and then intervened by medication (treated with CM complex prescription of treatment from lung, CM complex prescription of treatment from intestine, and SASP). After intragastric administration for 4 weeks, rats were sacrificed and samples taken. The expression of tumor necrosis factor α (TNF-α) and IL-8 contents in the lung tissue, the intestinal tissue, and the serum were detected by radioimmunoassay. Serum MedCAM-1 contents were detected using ELISA. Changes of the expression of Toll-like receptor 4 (TLR4), nuclear factor κB (NF-κB), neutrophil migration inhibition factor (MIF), mucosal addressin cell adhesion molecule-1 (MadCAM-1) mRNA in the lung tissue and the intestinal tissue were detected by real time PCR.

RESULTSCompared with the normal control group, the expression levels of TNF-α, TLR4 mRNA, IL-8, MIF mR- NA, and MadCAM-1 mRNA obviously increased in the model group (P < 0.01). Compared with the model group, the expression levels of TNF-α, TLR4 mRNA, IL-8, MIF mRNA, and MadCAM-1 mRNA obviously decreased in the treatment 1 and 2 groups (P < 0.01). The expression of MadCAM-1 mRNA in the intestinal tissue was obviously higher in the model group than in the normal control group (P < 0.01), while the expressions of TNF-α and NF-κB mRNA was obviously lower in the model group than in the normal control group (P < 0.05, P < 0.01). Compared with the model group, the expression of MadCAM-1 mRNA all significantly deceased in each treatment group (P < 0.05, P < 0.01). Serum TNF-α contents were higher in the model group than in the normal control group (P < 0.05). Compared with the model group, serum TNF-α contents could be lowered in the treatment 1 and 2 groups (P < 0.05, P < 0.01).

CONCLUSIONSThe main mechanisms of the intestinal injury in this UC model might be related with activation of acquired immune response, accompanied with lowered functions of inherent immune response. The main mechanisms of the lung injury in this UC model might be related acquired immune response and inherent immune response. Treatment from Fei and treatment from Dachang both could obviously improve the immunodissonance of Fei and Dachang, indicating the special relation between the lung tissue and the intestinal tissue, thus providing experimental evidence for Chinese medical theory of Fei and Dachang being interior-exteriorly correlated.

Aleurites ; Animals ; Colitis, Ulcerative ; drug therapy ; immunology ; Drugs, Chinese Herbal ; therapeutic use ; Enema ; Interleukin-8 ; metabolism ; Intestinal Mucosa ; immunology ; Intestines ; immunology ; Lung ; immunology ; Lung Injury ; NF-kappa B ; metabolism ; RNA, Messenger ; Rats ; Tumor Necrosis Factor-alpha ; metabolism

The purpose of this study was to explore the significance of abnormal karyotype in diagnosis and prognosis estimation of myelodysplastic syndrome (MDS). Chromosome analysis were performed in 306 cases of MDS using the short-term culture of bone marrow cell and G-banding technique, and in partial cases FISH technique was used for this analysis. 93 out of 306 cases were followed up. The results showed that 144 cases (47.1%) had clonal chromosome aberrations. The most common chromosomal aberrations included +8, translocation, complex or high complex karyotype, -7/7q-, 20q-/-20, trisomy 1 or partial trisomy 1, +11/+11q-, -9/9q-, +9/9q+, -Y, dup(1q), +21. The rate of abnormal karyotype in refractory anemia with erythroblasts (RAEB) and refractory anemia with erythroblasts-transformation (RAEBT) were much higher than in refractory anemia (RA) and refractory anemia with sideroblasts (RAS) (P < 0.05). The rate of abnormal karyotype among those cases with mutagen contact history were higher than those in cases without mutagen contact history. The patients with abnormal karyotype had a mean survival time much shorter than patients with normal karyotype (P < 0.005) and had a higher risk transforming into acute leukemia (P < 0.05). The worst outcome was observed in those patients with a complex or high complex karyotype, -7/7q- and trisomy 11. In conclusion, MDS is highly heterogeneous disorders and karyotype analysis is helpful for its diagnosis, treatment selection and prognosis estimation.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Chromosome Aberrations ; Female ; Humans ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; mortality ; Prognosis

Objective To explore a kind of simple and high efficient approach to differentiate human embryonic stem cells(hESCs)into neural stem cells(NSCs).Methods hESCs were cultured in bacterial culture dish filled with serum free medium to gain embryoid bodies.Then the mature embryoid bodies grew in the special medium including B27 and noggin by adherent culture to differentiate into NSCs. Results The hESCs kept floating in the bacterial culture dish and growing all the time and then formed mature embryoid bodies 7 to 10 days later.The embryoid bodies could be differentiated into highly pure (96.4%)nestin positive cells.And these cells were differentiated into all kinds of neural cells if cultured further.Conclusions This kind of method is less time-consuming,cheaper,and more efficient than those of the results in literatures reported.It affords very good source of seed cells for cell transplantation therapy in the future.

BACKGROUNDAhmed glaucoma valves (AGV) has been used for decades, but there is no detailed report about the efficacy of AGV in Chinese glaucoma patients. This study aimed to compare the intraocular pressure (IOP) lowering efficacy and side effects of S-2 polypropylene and PF-7 silicone AGV implantation in Chinese refractory glaucoma patients.

METHODSPatients were divided into S-2 model AGV group and FP-7 model AGV group. The complete and qualified surgical success rate, change of IOP, number of anti-glaucoma medications used and postoperative complications were recorded and analyzed.

RESULTSAverage follow-up time was comparable between two groups. IOP was reduced from (37.9 ± 12.7) mmHg preoperatively to (17.3 ± 5.3) mmHg at the last follow-up in S-2 group and reduced from (39.9 ± 14.4) mmHg to (17.7 ± 4.9) mmHg in FP-7 group. Anti-glaucoma medications were reduced from 3.8 ± 0.2 to 1.5 ± 0.2 in S-2 group, and 3.5 ± 0.2 to 0.7 ± 0.2 in FP-7 groups. The cumulative success rates were comparable in two groups, which were 61.2% and 72.1% in S-2 group and FP-7 group respectively. When IOP reduction criteria was used, complete success rates were 30.6% and 51.2% for S-2 and FP-7 groups, and qualified success rates were 86.1% and 92.7% separately. In both groups, the major complication was hypotony, and the previous trabeculectomy of patients was the major risk factor for surgery failure.

CONCLUSIONSIn this short-term retrospective study, S-2 AGV is showed at least as effective as FP-7 AGV in IOP reduction, but associated with higher rate of complications. Previous trabeculectomy is a principle risk factor for AGV implantation failure. These clinical outcomes are important for converting use of the FP-7 silicon AGV in Chinese refractory glaucoma patients.

Adult ; Female ; Glaucoma ; physiopathology ; therapy ; Glaucoma Drainage Implants ; Humans ; Intraocular Pressure ; physiology ; Male ; Retrospective Studies

OBJECTIVETo investigate the influence of a lipopolysaccharide receptor CD14-159C/T genomic polymorphism on CD14 gene expression as well as protein release, and the relation of sepsis susceptibility and prognosis in patients with extensive burns.

METHODSThe study group consisted of 26 patients with burns covering more than 30% of the total body surface area. The CD14 gene polymorphism was determined by polymerase chain reaction (PCR) and subsequent HaeIII restriction enzyme digestion of the PCR products. Meanwhile, the association of CD14, TNF-alpha mRNA expression in leukocytes and soluble CD14 (sCD14) levels in serum with CD14-159 polymorphism as well as prognosis after burns was also studied.

RESULTSThe T allele frequencies in sepsis patients and non-survivors were higher than those in non-septic patients and survivors. The levels of CD14 mRNA, TNF-alpha mRNA expression and serum sCD14 were significantly different among patient groups with TT, TC, and CC genotypes. The above differences were also existed between survivors and non-survivors. CD14 mRNA expression was higher in heterozygotes (TT and TC) than in C homozygous patients (P < 0.05 or P < 0.01), and sCD14 level was higher in heterozygotes (TC) than C homozygous patients on day 7 postburn (P < 0.05). During the 28-day observation period, mean TNF-alpha mRNA expression was higher in patients homozygous for T allele than C homozygotes (P < 0.05). In addition, higher CD14 mRNA values were found in non-survivors compared to those in survivors on days 7 and 28 postburn (P < 0.05).

CONCLUSIONSCD14C-159T polymorphism might markedly influence CD14 mRNA expression and sCD14 levels, and it seems to be associated with sepsis susceptibility and prognosis in patients with extensive burns. The T allele could be a genetic risk marker of adverse prognosis.

Adult ; Burns ; complications ; metabolism ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Lipopolysaccharide Receptors ; biosynthesis ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Prognosis ; RNA, Messenger ; genetics ; Sepsis ; etiology ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

OBJECTIVETo observe the effect of different concentration of Tamoxifen ointment on the fibroblasts and transforming growth factor (TGF-beta2) of hypertrophic scar at rabbit ears, so as to explore the possibility of treatment of hypertrophic scar with Tamoxifen.

METHODSThe hypertrophic scar model was established in 96 New Zealand rabbits' ears. The wounds were divided into four groups (A, B, C and D), with 144 wounds in each group. Different concentration of tamoxifen ointment (0.5%, 1%, 2%) was topically administered in groups A, B and C respectively, and blank ointment in group D. On postoperative day 30, 60 and 90, the scar samples were harvested. The scar thickness, scar histological change and the content of TGF-beta2 were detected.

RESULTS(1) On the 30th day after operation, the difference of scar tissue thickness among groups A, D and B, C reached statistical significance (group A, D < group B < group C). However, there was a contrary tendency in fibroblasts density and TGF-beta2 content of the scar tissue simultaneously. (2) On 60th, 90th day after injury, there was statistical difference in scar thickness, fibroblasts density and the content of TGF-beta2 in scar of four groups (P < 0.05). The content of TGF-beta2 in group A, B, C, D was (43.97 +/- 3.63) microg/L, (41.92 +/- 3.91) microg/L, (36.69 +/- 4.15) microg/L, (54.90 +/- 4.71) microg/L, respectively, on 60th day; and (45.69 +/- 2.63) microg/L, (40.43 +/- 3.87) microg/L, (38.76 +/- 3.24) microg/L, (52.59 +/- 4.92) microg/L, respectively, on 90th day. The fibroblasts density of scar in groups A, B, C, D was (4392.07 +/- 327.84) point/mm2, (4208.57 +/- 329.76) point/mm2 (4 033.44 +/- 427.91) point/mm2, (4863.03 +/- 387.98) point/mm2, respectively, on 60th day; and (4418.41 +/- 432.52) point/mm2, (4077.65 +/- 386.70) point/mm2, (3844.53 +/- 354.29) point/mm2, (4838.64 +/- 390.52) point/mm2, respectively, on 90th day. The content of TGF-beta2 and fibroblasts density of scar were lined up as group D > group A > group B > group C (P < 0.05).

CONCLUSIONSTopical Tamoxifen can reduce the content of TGF-beta2 and fibroblast, decrease fibroblasts density and the formation of hypertrophic scar at rabbit ears. It offers a new way for the treatment of the hypertrophic scar.

Animals ; Cicatrix, Hypertrophic ; drug therapy ; metabolism ; pathology ; Disease Models, Animal ; Ear Diseases ; drug therapy ; metabolism ; pathology ; Fibroblasts ; drug effects ; pathology ; Ointments ; Rabbits ; Tamoxifen ; pharmacology ; Transforming Growth Factor beta2 ; metabolism

To combine fingerprint and drug release rate in vitro, in order to study in vitro release of complex components of Chuanping sustained tablets, compound traditional Chinese medicine preparation. A qualitative determination of the characteristic peaks of the compound preparations were conducted by the fingerprint. The results of the dissolution rate determination under different release conditions showed that the release of three index components (methamphetamine, pseudoephedrine and scopolamine) of Chuanping sustained tablets was less affected by gastrointestinal factors, with similarity factors being more than 80 with unknown component release curves of three major characteristic peaks in the fingerprint. The qualitative determination proved that multiple components of the compound traditional Chinese medicine preparation was dissolved in vitro at similar rates, realizing the balanced release of complex components of the compound traditional Chinese medicine preparation. This study layed a theoretical and experimental basis for quality evaluation for the compound traditional Chinese medicine preparation.
Drugs, Chinese Herbal ; chemistry ; Tablets

The aim of this study was to investigate the expression status of the helicase antigen (HAGE) transcript and its clinical significance in patients with acute myeloid leukemia (AML) and chronic myeloid leukemia (CML). The expression of HAGE cDNA in bone marrow mononuclear cells from AML and CML patients was detected by using real-time quantitative PCR. The results indicated that overexpression of HAGE transcript (117.12% - 9842.70%, median 434.96%) was detected in 14.8% (11/74) AML patients. AML patients with HAGE cDNA expression were significantly older than those HAGE-negative patients (median 67 and 45 years, respectively, p = 0.001). HAGE cDNA expression was more frequently present among the patients with acute monoblastic leukemia (M(4) and M(5), 7 of 20, 35.0%), compared to the patients with acute non-monoblastic leukemia (M(1), M(2), M(3) and M(6), 4 of 54, 7.4%) (p = 0.007). 28.6% (8/28) cases with normal karyotypes showed HAGE cDNA overexpression, significantly higher than 7.5% (3 of 40) in those with chromosomal abnormalities (p = 0.041). Overexpression of HAGE transcript was found in 9 (34.6%) CML cases and more frequently observed at accelerated phase and blast crisis (4/4, 100%) than that at chronic phase (5/22, 22.7%) (p = 0.008). It is concluded that HAGE cDNA expression is relevant to specific subtypes of AML and to the progression of CML.
Blast Crisis ; Bone Marrow Cells ; metabolism ; DEAD-box RNA Helicases ; genetics ; metabolism ; DNA, Complementary ; Gene Expression ; Humans ; Karyotype ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics

OBJECTIVETo explore the mechanism of acupuncture on the treatment of glaucoma so as to find out a kind of simple and effect approach to control and stabilize intraocular pressure (IOP) of patients with glaucoma.

METHODSThirty-nine cases (75 eyes) of glaucoma (unstable control of IOP) were treated with acupuncture at Sibai (ST 2), Cuanzhu (BL 2), Jingming (BL 1), Qiuhou (EX-HN 7), etc. IOP before and after acupuncture, at 10 a.m, 2 p.m, 6 p.m, 10 p.m., at 5 a.m. and 7 a.m. on the second day as well as diurnal IOP variation in 24 h were observed separately.

RESULTSAfter acupuncture, IOP at each time point reduced as compared with that before acupuncture. IOP at 5 a.m., 7 a.m., 10 a.m., 6 p.m. and 10 p.m. after acupuncture was different significantly in statistics as compared with that before acupuncture correspondingly (all P<0.05). After acupuncture, 24 h diurnal IOP variation was (5.31 +/- 2.84) mmHg, which was lower significantly than (7.06 +/- 3.86) mmHg before acupuncture (P<0.05).

CONCLUSIONFor the patients with unstable control of IOP, acupuncture can not only reduce IOP, but also stabilize 24 h diurnal IOP variation and benefit the visual function of patients with glaucoma.

Acupuncture Therapy ; Adult ; Aged ; Female ; Glaucoma ; physiopathology ; therapy ; Humans ; Intraocular Pressure ; Male ; Middle Aged