Objective To study the prenatal genetic diagnostic methods for hemophilia A fetus.Methods From 2002 to 2006,19 hemophilia A families were diagnosed either by long distance-polymerase chain reaction(LD-PCR)for factor Ⅷ intron 22 inversion or by the DNA polymorphism genetic linkage analysis of factorⅧin the Beijing Chaoyang Hospital.Results (1)Totally 19 women,with 22pregnancies received the prenatal diagnosis of fetal hemophilia A.The average week at diagnosis was 23(17-34)weeks.All the direct fetal blood sampling(DFBS)were successful.There was no fetal-loss caused by the procedures.(2)Of the 19 hemophilia A families,14 appeared to be factorⅧintron 22inversion,in which 16 prenatal diagnoses were done,10 fetuses were diagnosed as genetical hemophilia A patients,and 6 fetuses were normal.(3)Using combined polymorphism genetic linkage analysis 6 prenatal diagnoses were done,including one woman's two pregnancies,in which both her fetuses were diagnosed as genetical hemophilia A patients.(4)Factor Ⅷ levels of 16 fetuses were measured,and 6 fetuses were unmeasured either because the pregnancy weeks were lower than 20 weeks or the parents refused.Factor Ⅷlevel ranged from 0 to 198%.There were 11 fetuses whose factor Ⅷ levels were lower than 10%.Ten of them were diagnosed to be genetical hemophilia A patients,and in only one boy the factorⅧlevel was 2%,but the genetic diagnosis was normal and for one year's follow up he was doing normal.Conclusion LDPCR combined with polymorphism genetic linkage analysis enables a quick and correct detection ofhemophilia A carrier.For a carrier pregnancy.prenatal diagnosis could be done for the male fetus.Factor Ⅷ deficiency of the fetus could help make the diagnosis but the final diagnosis should be based on genetic evidence.

Objective To investigate the effect of costimulatory molecules 4-1BB/4-1BBL on T cell activation with systemic lupus erythematosus(SLE)patients.Methods The expression of NF-kB mRNA and p38 MAPK mRNA of T cells with 20 SLE patients and 20 normal controls before activation,after activation and blocked bv 4-1BB/4-1BBL were detected bv RT-PCR.The protein levels of p-p38 MAPK and NF-kB were detected by Western blot.Results The expression of NF-kB mRNA and p38 MAPK mRNA and the protein expression of NF-kB and p-p38 MAPK of T cell with SLE patients were higher than that of normal controls(P＜0.01).There were more expression of NF-kB mRNA,p38 MAPK mRNA,NF-kB protein and p-p38 MAPK protein of T cell with SLE patients stimulated by anti-CD3 monoantibody(P＜0.01).In anti-4-1BB monoantibody blockage group,there were decreased expression of p38 MAPK mRNA and p-p38 MAPK protein(P＜0.01).but not the decreased NF-kB mRNA and protein expression.Conclusion Costimulatory molecule 4-1BB plays a critical role in T cell activation of SLE by p38 MAPK signal pathway.

Animals ; Carcinoma, Squamous Cell ; genetics ; Colorectal Neoplasms ; genetics ; metabolism ; Enzyme Activation ; Esophageal Neoplasms ; genetics ; Genome-Wide Association Study ; Head and Neck Neoplasms ; genetics ; Humans ; Neoplasms ; chemically induced ; enzymology ; genetics ; Phosphoinositide Phospholipase C ; chemistry ; genetics ; metabolism ; physiology ; Signal Transduction ; Skin Neoplasms ; chemically induced ; enzymology ; Stomach Neoplasms ; genetics ; Urinary Bladder Neoplasms ; metabolism ; pathology ; ras Proteins ; metabolism

Adenocarcinoma, Mucinous ; pathology ; Breast Neoplasms ; pathology ; Carcinoma, Ductal, Breast ; pathology ; Carcinoma, Renal Cell ; pathology ; Carcinoma, Squamous Cell ; pathology ; Cystadenocarcinoma, Mucinous ; pathology ; Cystadenoma, Mucinous ; pathology ; Female ; Giant Cells ; pathology ; Humans ; Osteoclasts ; pathology ; Ovarian Neoplasms ; pathology ; Pancreatic Neoplasms ; pathology ; Thyroid Carcinoma, Anaplastic ; Thyroid Neoplasms ; pathology ; Tongue Neoplasms ; pathology ; Urologic Neoplasms ; pathology

Objective To select the optimum extracting method of polysaccharide from Paris polyphylla Smith var. chinensis(Franch. )Hara in Shaan'xi. Methods The optimum extraction technology and condition were determined by using response surface methodology and single factor experiment,with the extraction ratio of polysaccharides from Paris polyphylla Smith var. chinensis( Franch. ) Hara in Shaan'xi as indicator. Results The optimum extraction condition for the polysaccharides was as follows:extracting at 92 ℃ for 2. 6 h and with the liquid to solid ratio of 15 ( mLg). The average extraction yield of polysaccharide was 3. 22%under the optimum extraction condition. Conclusion The method is suitable for the extraction and determination of polysaccharides from Paris polyphylla Smith var. chinensis( Franch. ) Hara in Shaan'xi.

Objective To explore the regulation mechanism of autophagy-related protein, microtubule-associated protein 1 light chain 3 (LC3), via c-Jun in methotrexate resistant human choriocarcinoma JEG-3 cell lines. Methods Human choriocarcinoma JEG-3 cell lines, and methotrexate resistant choriocarcinoma JEG-3 (JEG-3/MTXR) cell lines were used in our present study. Phosphorylation c-Jun (p-c-Jun) was evaluated after exposure to 0.02 ng/ml methotrexate for 72 hours in both cells by western blot. c-Jun gene was knockdown by small interference RNA (siRNA) in JEG-3/MTXR cells, and LC3 was evaluated by western blot and reverse transcription-PCR. The binding of LC3 promoter with c-Jun protein was detected via chromatin immunoprecipitation assay (ChIP) with or without 0.02 ng/ml methotrexate exposure. Results The results showed that p-c-Jun was up-regulated after methotrexate treatment for 72 hours (1.99±0.20, versus 0.20±0.06 at 0 hour;P<0.05) by western blot analysis in JEG-3/MTXR cell lines. Further investigation demonstrated that c-Jun-siRNA could inhibit the up-regulation of LC3 formation and after methotrexate exposure (LC3 mRNA:1.24±0.17 versus 3.03±0.43;LC3 protein:0.52±0.07 verus 1.20± 0.15; all P<0.05). The binding of LC3 promoter by c-Jun protein was up-regulated after methotrexate treatment by the method of ChIP in methotrexate resistant JEG-3/MTXR cells [(2.95 ± 0.35) times]. Conclusion Autophagy-related gene LC3 expression regulated by c-Jun protein may be involved in the effect mechanism of the development of methotrexate resistance in choriocarcinoma JEG-3 cells.

Objective To investigate the expression level of miRNA-146a (miR-146a) in retinal pigment epithelial (RPE) cells aging and age-related macular degeneration (AMD),and discuss its regulation mechanism of AMD by repressing VEGF-A.Methods The expressions of miR-146 and VEGF-A were examined by qRT-PCR in RPE cells in mice aged 2 months,8 months,12 months,18 months or 24 months,and in RPE cells from 75 years old AMD patients.The protein level of VEGF-A was also detected by Western Blotting.Finally,the effects of overexpression of miR-146a in APRE-19 cell line on expression of VEGF-A was checked.Results MiR-146 was up-regulated to 8 times or 24 times at 18 months or 24 months aged mice,and the expression of VEGFA was down-regulated in aging RPE from 1.5 times to 0.8 times.However,the expression of miR-146 decreased to 14.5 times and VEGF-A increased in RPE cells of AMD.In cultured cells,overexpression of miR-146a inhibited the expression of VEGF-A.Conclusion Up-regulation of miR-146a in aging RPE cells and its down-regulation in AMD suggest a potential of miR-146a as molecular marker.MiR-146a overexpression inhibits the expression of VEGF-A,supporting a potential clinical treatment of miR-146 in AMD.

Background Vitretomy and lenstomy with silicone oil tamponade is an effective method for complicated vitreous retinopathy.The severe anisometropia after silicone retention is usually treated by two-point transscleral suture fixation for posterior chamber intraocular lens(IOL)implantation.In order to reduce the number and difficulty and complication of the operation,the surgical method should be improved.Objective The goal of this study was to observe the resuh of silicone oil removal combined with four-point trans-scleral suture fixation of posterior chamber IOL after vitrectomy.Methods A retrospective case-observational study design was adopted.Twenty eyes with silicone oil tamponade from 20 patients without lens and posterior capsule after vitrectomy were included in this study.Silicone oil removal with four-point trans-scleral suture fixation of posterior chamber IOL was performed.The anterior ocular inflammatory response,intraocular pressure,uncorrected and corrected acuities before and after operation,corneal endothelial cell counting and postoperative complications were observed and analyzed.Written informed consent was obtained from all patients prior to the operation.Results All of the operative eyes in this study showed improvement of visual acuity after operation.Of the 20 eyes,a visual acuity of ≥0.8 was seen in 2 eyes,0.6-0.7 in 6 eyes,0.3-0.5 in 8 eyes and 0.05-0.2 in 4 eyes 3 months after the removal of silicone oil.The uncorrected acuity postoperation was significantly improved in comparison with preoperation(H=10.147,P＜0.01),but no significant difference was found in the corrected acuity between preoperation and postoperation(X =2.089,P＜ 0.01).The number of the corneal endothelial cells was(2064±329)cells/mm2 before operation,and that after operation was(1987±269)cells/mm2,showing an insignificant change between them(t =1.660,P ＞ 0.05).No abnormality of IOL position was found in all 20 operated eyes.There was not serious postoperative complication in all 20 patients.Conclusions The combination of silicon oil extraction with four-point transscleral suture fixation IOL is effective in eyes without posterior capsule or lens after vitrectomy.It can reduce the operation time and improve the postoperative acuity and the quality of life of patients.

Objective To evaluate the protective effect of leukocyte-depleted warm blood cardioplegia on immature myocardium in infant.Methods Thirty infants with congenital heart disease whose American Society of Anesthesiologists(ASA) were Ⅱ-Ⅲ,aortic clamping time and bypass time were more than 30,40 minutes were respectively selected and divided into 2 groups:Experimental group and control group.Experimental group were perfused with leukocyte-depleted warm blood cardioplegia while control group perfused with common warm blood cardiplegia.Under monitoring the hemodynamics at surgery,the serum levels of troponin I(cTnI) and intercellular adhesion molecule-1(ICAM-1) in heparinized anticoagulant blood samples from radial artery at different time points [anesthesia induction and before extracorporeal circulation(T1),30 min after aortic clamping(T2) and aortic declamping 5 min(T3),15 min(T4)]were detected.Three pieces of cardiac muscle were taken from right atrium at different time points and the levels of myeloperoxidase(MPO) were detected.Results 1.The serum cTnI and ICAM-1 levels after aortic declamping were higher than those before operation,and the levels in experimental group were lower than those in control group(t=2.358,2.533,2.30,2.639 Pa

Objective To explore the diagnosis and curative effect of phenylketonuria(PKU) in Gansu.Methods Fifty-nine thousand nine hundred infants and 118 borderline cases were screened from Oct.1999 to Dec.2006.Three drops of venous blood was collected from the exterior or interior part in the heel,and trickled the venous blood onto the specialized filter paper,which could be formed into 3 bloodstains with the diameter of 0.8-1.0 cm.Phenylalanine(phe) was detected by chemiluminescence method,and PKU was screened by the PKU standard of diagnosis.Infants with PKU had been given low phe food.During the treatment,phe level,physical development and intellectual development were measured regularly.Patients′ body weight,height and head circumference were measured once 1 month.Patients′ intellectual quotient(IQ) were measured every half year(IQ was analyzed by Gesell test).Results were analyzed by SPSS 11.0 software for t test.Results In total,93 patients with PKU were diagnosed and treated.Thirty-six cases were identified by neonatal screening before 3 mouths old.Mental retardation was found in 57 cases before 6 years old.After treatment with low-phe diet,the follow-up for early-treated patients revealed that their physical and mental development were normal,and height,weight and head circumference were in the normal amplitude,and the results of IQ was(92?12)scores.In later treated patients,abnormal behaviors were significantly improved and their developmental quotients were elevated.Before treatment,Gesell value was(57?20)scores;after 12-18 months treatment Gesell value was(70?20)scores.The difference was significant(t=1.705 P