[Objective] To observe the therapeutic effect of pressing- moxibustion on Baihui (DU20) point combined with electroacupuncture (EA) on cervical spondylosis (CS) of vertebral artery type. [Methods] A single - blind random - controll trial was carried out in 34 cases of CS of vertebral artery type. The cases were randomized into two groups: the treatment group (Group A, n = 17) treated with pressing- moxibustion on Baihui point and EA on bilateral Fengchi (GB20) point and Jiaji (EX - B2) point of impaired vertebrae and the control group (Gropup B, n = 17) with EA alone. Plasma contents of thromboxane 82 (TXB2), 6-keto-prostaglandin Fl alpha (6-keto-PGF1?) and their ratio were measured before and after treatment. [Results] In the treatment group, 9 were cured, 6 markedly effective, 2 effective and 0 ineffective and 4,6,5 and 2 in the control group respectively; the markedly effective rate was 88.2% which was superior to that (58.8%) in the control group (P

Objective To explore the effect of bellybutton exposure on premature navel nursing effect.Method Three hundred and ninety-three premature infants were randomly divided into experiment group(n=195)and control group(198). Bellybutton fasciation and bellybutton exposure were used in the control group and the experiment group respectively. The two groups were compared in terms of umbilical shedding,time of umbilical healing,umbilical swelling,bleeding and breathing.Result The umbilical healing time in the experiment group was significantly shorter than that of the control group(P<0.05). The incidences of omphalitis,umbilical swelling and bleeding, changed breathing in the experiment group were significantly lower than those of the control group(all P<0.05).Conclusion The bellybutton exposure is effective in promoting the umbilical shedding in premature infants and reducing the incidence rate of premature navel infection.

Objective To investigate a method to detect the sentinel lymph nodes(SN) of thyroid papillary carcinoma and its predictive value for cervical metastasis of carcinoma. Methods Intraoperative methylene blue dye mapping was performed in 24 cases of thyroid papillary carcinoma. The coincidence rate of frozen section pathology and routine section pathology of SN was observed,and the predictive value of SN for (metastasis) of the cervical lymph nodes was noted. Results SN was successfully detected in 21(87.5%) of 24 cases. The average number of SN was 3 nodes. There was one false negative case, the false negative rate was 4.8%(1/24), and no false positive cases were found. The predictive value of sentinel lymph nodes to (cervical) lymph node metastasis was 83.3%.Conclusions Methylene blue staining to identify sentinel lymph nodes could accurately predict the status of cervical lymph node metastasis of thyroid papillary carcinoma.

Objective To study the gene expression profiles of megakaryocytes(MKs) from human cord blood CD34+ cells in vitro expanded and to understand megakaryopoiesis at the molecular level. Methods CD34+ cells were isolated using density gradient centrifugation and magnetic activated cell sorting. The cells were cultured and stimulated with recombinant human TPO ( 100 ng/ml). After 12 days, the MKs fraction was separated using an anti-CD41 monoclonal antibody by immunomagnetic sorting. The gene expression profiles of MKs, non-MKs as well as meg-01 cells were studied by gene chip assay. THBSI, HOX A9,β-actin, lL-8,Annexin A6, FGF-8 were selected to validate the gene chip results by RT-PCR. Results A total of 116 genes between MKs and non-MKs cells were significantly different, 52 genes were up-regulated and 64 genes were down-regulated. In addition, 158 genes between MKs and meg-01 cells were significantly different, 71 genes were up-regulated and 87 genes were down-regulated. THBSI showed higher expression in MKs than in non-MKs. HOXA9 showed lower expression in MKs than in non-MKs. The expression of β-actin did not show any significant difference in MKs and non-MKs. IL-8 showed higher expression in MKs than in meg-01 cells, while ANXA6 showed lower expression in MKs than in meg-01 cells. The expression of FGF-8 did not show any significant difference between MKs and meg-01 cells. Conclusions MKs, non-MKs and meg-01 cells show different gene expression profiles. The regulatory genes include stress response genes,immune related genes, DNA synthesis and repair genes, metabolism genes, pro-onco genes and tumor suppressor genes.

AIM: To explore whether AT2 receptor is expressed in skeletal muscular vasculature, and the expression of angiotensin Ⅱ receptors in pulmonary circulation of children with left-to-right shunt but without obstructive pulmonary hypertension. METHODS: Lung and skeletal muscular tissue were obtained from 20 children with left-to-right shunt by biopsy during operation. These skeletal muscular tissues were detected by reverse transcriptase/polymerase chain reaction (RT-PCR ) and immunohistochemistry techniques for AT2 receptors. mRNA of AT1 and AT2 receptors in lung tissues were detected by RT-PCR and analyzed semi-quantitatively. RESULTS: In all of the skeletal muscular tissues, mRNA of AT2 receptor was found by RT-PCR, and the results of immunohistochemistry staining for AT2 receptor of vessels were positive. In the lung tissues, the level of mRNA of AT2 receptor was different to AT1 receptor, and the former was higher than the latter (P

Objective To analyze the molecular genetic basis of novel allele HLA-B * 9534 and establish the allele group specific primer PCR method. Methods Genomic DNA was extracted from whole blood by commercial DNA extraction kit. The HLA-B exons 1 to 8 coding sequences of the proband were am-plified by PCR and the amplification product was purified with double enzymes digestion and both strands of exons 2, 3 and 4 were sequenced. The exon 2-4 amplification of the HLA-B * 9534 was performed with al-lele group specific primers PCR and the PCR product was directly sequenced for exon 2 to 4. Results The proband has two HLA-B alleles. The result was assigned for HLA-B * 1518 and B * 4601 combination with a mismatch in 593A/G heterozygote by DNA sequencing of exon 2 to 4 with loci primers. After separating the two alleles of the proband with allele group specific primers polymerase chain reaction method, HLA-B * 4601 and HLA-B * 9534 alleles were identified after sequencing. The HLA-B * 9534 is identical to HLA-B * 1518 except for one nucleotide substitutions in exon 3 at position 593 A→G, this results in amino acid substitution at cedon 174 from Asn to Ser. The sequences of the novel allele have been submitted to GenBank (EU046491) and the allele has been officially nominated by the WHO Nomenclature Committee. Conclusion Identification of a novel HLA-B * 9534 allele and allele group specific primer PCR for HLA-B * 9534 was re-liable.

Objective To investigate the serological characteristics and molecular basis of the B (A)phenotype in ABO blood group and provide the data for clinical transfusion of individuals with B(A) phenotype.Methods The ABO group antigens on red cells of the proband,family members and donors were identified by monoclonal antibodies and the ABO antibodies in sera were detected by the standard A,B,O cells.The compatibility testing for the proband and donors was detected by salted test,polybrene test and antiglobulin test.The coding region of exon 6 to exon 7 in ABO gene was amplified by polymerase chain reaction(PCR) and the PCR products were sequenced.The haplotypes of proband were analyzed by cloning and sequencing.Results It was showed that both A and B antigens were detected on red cells of the proband and her two family members,and there was anti-A_1 antibody in their sera.The serological phenotype of the samples are identified as the A_2B.DNA sequencing showed 261 G/del,297A/G,526C/G,657C/T,700C/G,703G/A,796C/A,803G/C,930G/A heterozygotes in exon 6 to exon 7.It can be deduced that genotype in the proband is B(A)_(02)/O_(01).The genotypes of her mother and grandmother-in-law were B(A)_(02)/B_(101) and B(A)_(02)/O_(01),respectively.After cloning and sequencing,two alleles B(A)_(02) and O_(01) in proband was showed.B(A)_(02) has snigle nucleotide change(700 C>G),which resets replacement of proline with alanine at position 234.Two donors with phenotype A_2B were identified as genotype B(A)_(02)/O_(01) and A_(208)/B_(101),respectively.The results of crossmatch testing is in accordane between the proband and two donors and there was no clinical adverse reaction after transfusion.Conclusions 700C>G in α-1,3galactosyltransferase allele(B allde)can result in B(A)phenotype in individuals with the phenotype of A_2B.The donors in the transfusion for the individuals with B(A) phenotype should include individuals with A_2B phenotype.

Objective To establish a PCR sequencing-based typing (PCR-SBT) method for simultaneous genotyping of human platelet antigen HPA-1 to HPA-16w.Methods All DNA polymorphism sites of HPA-1 to HPA-16w were obtained from the immuno polymorphism database.The specific primers were designed using Primer Premier 5.0 software to amplify nucleotide acid fragments encompassing each HPA polymorphism site.The primer sequence and PCR condition were optimized to obtain specific and single amplification product.The PCR product was purified and then sequenced to determine the HPA genotypes.Two standard DNA samples were detected using the HPA PCR-SBT method to examine the accuracy d this method.Sixteen reference samples (including 6 interference samples with HPA gene mutations) provided by 14th platelet immunology workshop of international society of blood transfusion (ISBT) in 2008 were also tested by this home-brew HPA PCR-SBT method.Results Total eleven pairs of primers were designed to amplify and sequence the sixteen HPA systems.The HPA genotypes of two standard samples were 1aa/2aa/3ab/4aa/5ab/6aa/7aa/8aa/9aa/10aa/11aa/12aa/13aa/14aa/15aa/16aa and 1aa/ 2aa/3aa/4aa/5aa/6aa/7aa/8aa/9aa/10aa/11aa/12aa/13aa/14aa/15aa/16aa,respectively.The 256 HPA genotypes of 16 reference samples were clear.128 genotypes among them were completely accordance with the results provided by ISBT report.Conclusions The PCR-SBT assay combining high-throughput DNA sequencer established in the study provides a simple,rapid and accurate method for HPA-1 to HPA-16w systems genotyping.The assay is suitable for routine clinical HPA genotyping and shows a broad prospect in further applications.

Objective To determine the effect of lipoxins (LX) A4 and its agonist (BML-111) on the survival of RAW264.7 macrophage cells and TLR4/NF-κB signaling pathway. Methods RAW264.7 cells were treated with different concentrations of LPS, then the effect of LX A4 and BML-111 on the survival rate of these cells was observed. Cytotoxicity were detected by CCK-8 method and RT-PCR was used to detect the TLR4 and TRAF6 mRNA. The protein levels of TLR4 and pNF-κB p65 in RAW264.7 were determined by Western Blot. Results The survival rates of macrophage treated with LPS for 6 h in 1 000 ng/mL LPS in LX A4 group and BML-111 group were significantly higher than that in control group (P < 0.05). In the present of LPS, the TLR4 mRNA levels in RAW264.7 cells from LX A4 group and BML-111 group were significantly higher than those in the corresponding non-LPS groups. And the TRAF6 mRNA levels in each LPS stimulation group were higher than those in the corresponding non-LPS groups (P<0.05), while the protein level of TRAF6 in LX A4 and BML-111 groups were significantly lower than that in control group (P < 0.05). Stimulated with LPS, the protein levels of pNF-κB p65 in the LX A4 group and BML-111 group were all significantly lower than that in control group (P<0.05), and pNF-κB p65 expression level in control group was also significantly higher than the corresponding non-LPS groups (P < 0.05). Meanwhile, no significant difference was found between LX A4 and BML-111 group (P > 0.05). Conclusion LX A4 and BML-111 could inhibit the cytotoxicity of LPS on the RAW264.7 macrophage cells through the inhibition of the activation of TLR4/NF-κB signaling pathway, then reduce the inflammation. And this stable BML-111 may appear as another promising treatment for IBD disease.

Objective To analyze the influence of diabetes mellitus (DM) on the outcomes of percutaneous transluminal angioplasty (PTA) with stenting of femoropopliteal artery,in patients with atherosclerotic occlusive disease (ASO).Method Clinical data of inpatients who successfully received PTA with stenting procedures of femoropopliteal artery in Xuanwu Hospital from January 2006 to December 2012 were analyzed retrospectively.Patients were stratified into DM and non-DM groups.Results were compared between the two groups including primary patency (PP),assisted patency (AP),limb salvage and survival using Kaplan-Meier life table and Cox regression analyses.Result Totally 291 patients underwent 332 procedures.There were 214 DM and 118 non-DM limbs.Mean follow-up was 34 months.The 5-year PP was 23.2％,AP was 35.3％,limb salvage was 89.2％,and survival was 69.6％.DM was associated with lower 5-year PP,limb salvage,and survival than non-DM.But there were no significant difference in AP between the two groups.Females were associated with decreased PP than males on Cox multivariate analysis.Hypertension and below tibial diseases were associated with lower limb salvage rate and age is the only predictor of survival rate.Conclusions DM is a risk factor for poor outcomes after PTA with stenting procedures.