OBJECTIVE:To determine the content of gallic acid in viviparous bistort rhizome by HPLC.METHODS:The chromatographic column was Nova-Pak C 18 and the mobile phase was0.2%methanol methyl cyanide solution-0.1%phos?phoric acid-0.15%triethylamin(3∶50∶47),the flow rate was1.0ml/min and the detection wavelength was271nm.RE?SULTS:It took a good linear relationship when the sample size of gallic acid was within the range of0.028～0.14?g(r=0.9991),the average recovery was99.05%(RSD=4.0%).CONCLUSION:The method is convenient,fast,reproducible,which can be used for the content determination of gallic acid in viviparous bistort rhizome.

Objective To study and observe the engraftment of donor cells from unrelated cord blood into adult patients with severe aplastic anemia (SAA) and the outcome of allo-CBSCT. Methods Six patients received cord blood provided by Guangzhou Cord Blood Bank, for three of which one unit of cord blood was given in a procedure, whereas for other 3 patients, 2 units of cord blood were infused at the same time in a transplant protocol. In all 9 units of umbilical cord blood (UCB) infused, UCB units contained 1.6 ～ 10.7 nucleated cells/kg body weight of the recipient after thawing. The patients were conditioned with decreased dosage of immunosuppressive agents of CTX (60?mg/kg) and ALG (120?mg/kg). The engraftment state of the donor cells into recipients was confirmed by microsatellite DNA fingerprinting and fluorescent quantitative PCR analysis. Results Engrafted evidence has been found in 5 patients by molecular biology analyses showing donor-recipient mixed chimerism post transplant which was stable and persistent. After a median follow-up of 21 months (range 7～50), 4 patients were alive and disease free. One patient died of severe infection in the 3rd month from transplant, though the evidence of engraftment was obvious. Another patient also died in the early stage posttransplant of serious aspergillus infection without the engrafted proof.Conclusion Durable donor-recipient stable mixed chimerism can be achieved by unrelated UCBT in patients with SAA. Umbilical cord blood could be employed as a source of hematopoietic stem cell for adult transplantation.

Objective To investigate the antimicrobial properties of human amniotic homogenate supernatant (HAHS) in order to found a theoretical base for the use of human amniotic membrane in ophthalmological field. MethodsFresh human amniotic membranes were used to make HAHS and acellular amniotic membranes. Then, we observed their antimicrobial effects and antimicrobial spectrums, compared the antimicrobial capacity with 10 commonly used antibiotics in eyes, and investigated the effects of time, temperature and pH value on the antimicrobial capacity. Finally, transmission electron microscopy (TEM) was used to explore the possible targeting site of the antibiosis. ResultsHuman amnion membrane contained antimicrobial components locating in its epithelial cells. HAHS had a broad antimicrobial spectrum and was steady in nature. Its antimicrobial capacity was stronger than those of sulfasulfonamide, chloromycetin and cefuroxime sodium. TEM indicated that the antimicrobial effect were exerted through plasma membrane of microorganism. ConclusionHAHS can be an effective and convenient treatment for ocular surface infectious diseases. Traditional amnion transplantation should employ fresh human amniotic membranes containing complete epithelial lamina to reconstruct the ocular surface.

Objective To explore the possibility of cytochrome P-450 3A5 (CYP3A5) genotype as a major factor to guide individualized employment of cyclosporin A(CsA) through a comparative study of CsA concentrations in the peripheral blood of hemopoietic stem cell transplant recipients with different CYP3A5 genotypes. Methods Olymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to genotype CYP3A5 gene, and CsA concentrations were detected by a commercial fluorescence polarization immunoassay. Result There were significant differences in CsA concentrations, including standardized trough concentrations C0 and two h peak concentrations C2 , in the two CYP3A5 genotypes found in our samples, and both C0 and C2 in wild homozygotes were lower than heterozygotes. Conclusion CYP3A5 polymorphism is highly associated with CsA concentrations in hemopoietic stem cell transplant recipients, and CYP3A5 genotype may be a predictor to the dose requirement before clinically employment of CsA.

A new method for noninvasive prenatal diagnosis of fetal sex was developed by using single-cell PEP-PCR techniques. Micromamipulation techniques were used to obtain single fetal cells from 273 maternal blood samples. The genome of single cells was preamplified by PEP and SRY genes were analyzed by PCR method. The SRY genes of 149 samples were detected by the new method among 153 samples carrying male fetus, while 119 out of 120 samples carrying female fetus were proved negative for SRY genes. The sensitivity and specificity of the new method were 97.39% and 99.17% respectively and the correct rate was 98.17%. The new method has the advantage of high sensitivity and specificity in noninvasive prenatal diagnosis of fetal sex and provides the basis of other researches such as sex-linked inherited diseases.
Chromosomes, Human, Y ; Erythroblasts/chemistry ; Fetus/*cytology ; Genes, sry/genetics ; Genetic Diseases, Inborn/diagnosis ; Maternal-Fetal Exchange/genetics ; Polymerase Chain Reaction/methods ; Pregnancy/*blood ; Prenatal Diagnosis/*methods ; *Sex Determination (Genetics)

Objective Pulmonary benign metastasizing leiomyoma (PBML) is a rare entity that leiomyoma of uterus metastasized to the lung.The clinical characteristics of this rare disease were analyzed in this article.Methods The detailed clinical records of 7 patients diagnosed as PBML at Peking Union Medical College Hospital between January 2001 and June 2015 were reviewed.Results All patients were women with median age of 44 years (range 28-62).Symptoms included dyspnea (2/7),chest pain (1/7),cyanosis (1/7),cough (1/7) and bloody sputum (1/7),while 4/7 cases were asymptomatic.Six patients had the past-history of leiomyoma of uterus 20 months to 14 years ago among whom 5 patients received hysterectomy.Chest CT showed bilateral,random-distributed multiple round solid nodules,or diffuse-distributed miliary nodules,or single solid nodule,even some small cavities.Extra-pulmonary metastasis was found in left superclavicular lymph node (1 case) and right heart (1 case).Histological tissues were obtained by video-assisted thoracic surgery lung biopsy (4/7),mass resection on tricuspid valve (1/7),transbronchil lung biopsy (1/7),and CT-guided percutaneous lung biopsy (1/7).Pathology showed an interlacing pattern by spindle cells having elongated nuclei without cellular atypia.Ki-67 index was less than 1％.Molecules such as smooth muscle antibody,estrogen receptor (ER) and progestrone receptor (PR) were positive in immunohistochemistry staining.Neither letrozole nor zoladex was effective.Two patients responded to bilateral adnexectomy,presenting as shrunk nodules.No relapsed disease was seen in one patient with single nodule after resection.There was only one patient with disease-related mortality,whose chest CT showed milliary nodules.Conclusion Although CT findings of PBML are similar to malignancies,the clinical outcome is good.Despite the positive expression of ER and PR,the effectiveness of hormone related treatment is limited.And periodical follow up is suggested even to those uneventful patients.

Objective In our previous work, the prevalence of anti-endothelial cell antibodies(AECA) in patients with systemic vasculitis and other autoimmune diseases was analyzed. AECA against a 47 000 endothelial cell antigen was found in patients of a variety of systemic vasculitis and systemic lupus erythematosus (SLE). It was suggested to be α-enolase by the combination of immunoblotting and proteomics methods. The aim of this work is to demonstrate that α-enolase is one of the targets of AECA, and to detect the prevalence of anti-α-enolase antibody in sera of patients with autoimmune disorders including systemic vasculitis. Methods The CDS of human Enol gene was amplified by polymerase chain reaction (PCR), with template of human placenta λzap express Cdna library. The product was then recombined with expression vector. After expression and purification from E.coli, the recombinant protein was analyzed by mass spee-trometry. The prevalence of anti-α-enolase antibody in patients with autoimmune disorders including systemic vasculitis was tested by Western blot and enzyme-linked immunosorbent assay (ELISA). Results The CDS of human Enol gene was subcloned to the expression vector. Recombinant human α-enolase was expressed and purified in E.coli. The recombinant protein was demonstrated to be his-tagged human a-enolase by mass spectrometry. Results of Dot-Blot revealed that the prevalence of anti-α-enolase antibody was 76.7% in systemic vasculitis [including 74.0% in Behcet's disease (BD), 81.5% in Takayasu artefitis (TA), 62.5% in Wegener's granulomatosus (WG), 92.3% in microscopic polyangitis (MPA) and 80.0% in Churg-Stranss syndrome (CSS)], 78.3% in SLE, 63.6% in Sjogren's syndrome (SS) and 78.9% in rheumatoid arthritis(RA). No positive signals were detected in sera of normal controls or patients with polymyositis/ dermatomyositis (PM/DM). There was no statistical significance among positive rates of anti-α-enolase antibody in systemic vasculitis, SLE, SS or RA patients. The prevalence of positive signals at the most extensive level (+++～++++) was 51.7% in patients with systemic vasculitis, 33.3% in SLE, 42.9% in SS and 20.0% in RA. There was statistical significant difference between RA and systemic vasculitis. Conclusion The identification of human α-enolase as one of the targets of AECA and its prevalence in a variety of autoimmune disorders will shed some light on the understanding of the pathogenesis of vascular injury in autoimmune diseases.

Objective To investigate the correlation between corneal neovascularization (CNV) in rat after cautery with acid and the defect in epithelium, the infiltrating of leucocytes. Methods The model of mild, moderate and severe acid burn in rat cornea was established. Corneal angiogenesis and the diameter of the epithelial defect were observed with a slit lamp microscope. The ratio of neovascularization area to that of cornea was calculated. The specimens were taken to stain with hematoxylin and eosin (HE) at different time points after cautery and the number of the leucocytes in cornea was calculated with light microscope. The relation between CNV and the diameter of epithelial defect and the infiltrating leucocytes in cornea was analyzed with Pearson's correlation. Results The healing of epithelium differed with different degree of acid burn. The more damage the eye suffered, the more time it would take to heal. Leucocytes infiltrated in cornea after cautery. The more damage the eye suffer, the more leucocytes can be observed, and the statistical difference was significant (P

Objective To evaluate the safety and efficacy of icotinib as first-line therapy in Chinese non-small cell lung cancer (NSCLC) patients harboring epidermal growth factor receptor (EGFR) sensitive mutations.Methods Patients with stage ⅢB/Ⅳ NSCLC who had EGFR sensitive mutation and had no previous treatment were enrolled into this study.The response rates , progress free survival ( PFS) , overall survival ( OS ) , and the safety were analyzed.Results Ninety advanced adenocarcinoma patients were enrolled in this study , 44 patients had partial response ( PR ) , 42 patients had stable disease ( SD ) , 4 patients had progressive disease (PD), with an overall response rate (ORR) of 48.9%, and a disease control rate (DCR) of 95.6%.The median PFS was 14.9 months (95%CI 13.5-16.3) and the OS was 37.0 weeks ( 95%CI 27.9 -46.1 ).Patients with brain metastases showed higher ORR ( P =0.049 ).Patients with stage ⅢB had longer PFS than those with stage Ⅳ( P=0.007 ).The most common adverse events were grade 1 -2 skin rash (38 patients, 40.9%).Other adverse events included dry skin , oral mucositis, diarrhea and liver function injury.Three patients withdrew because of severe liver injury or skin rash.No treatment related mortality occurred .Conclusions Icotinib is effective and safe as first-line treatment for Chinese advanced NSCLC patients with EGFR sensitive mutation.

Objective To identify and quantitate the conjunctival aerobic bacterial flora from healthy people of 3 age groups in Chongqing city.Methods Samples taken with moist swabs from the lower fornix of both eyes of 33 children(9.85?0.57 years old),31 youths(21.23?0.88 years old)and 30 elders(72.97?4.94 years old)were given bacterial cultures.Positive ones were identified and colony forming units(CFUs)were counted.Bacterial quantity was expressed as the number of CFUs divided by the weight of each sample measured before and after collection.Results The elders showed a higher isolation frequency(93.33%)than children(45.45%)and youths(50.00%)which had statistically difference(P