OBJECTIVETo optimize the therapeutic programs for periarthritis of shoulder treated with acupuncture, moxibustion and kinetohterapy with orthogonal design method adopted.

METHODSThe orthogonal design table of L8 (2(7)) hierarchical principle was used to randomly divide 192 patients of periarthritis of shoulder into 8 groups, 24 cases in each one. Separately, 4 factors and each different 2 levels were adopted in treatment, named acupuncture timing (factor A: A, acute stage, A2 adhesion stage), acupoint combination (factor B: B, local acupoints, B2 local acupoints and distal acupoints along meridians), filiform needling and warm needling therapy (factor C: C1 acupuncture with filiform needle, CZ acupuncture with filiform needle and warm needling therapy) and positive functional exercise (factor D: D1 without positive functional exercise, D2 with positive functional exercise). The treatment was given once a day, 10 treatments made one session and 2 sessions were required totally. The time points of observation were the point after 1 session of treatment and after 2 sessions of treatment. The short-form McGill pain questionnaire (MPQ) and shoulder joint motor disturbance score were adopted for evaluation.

RESULTSIn the orthogonal design analysis, taking the hierarchical factors into consideration, the age was considered as the main factor in the evaluation of shoulder pain and shoulder motor disturbance (P<0.01), and the shoulder function grade apparently impacted pain evaluation and the efficacy on shoulder motor disturbance (P<0.01). The best combination of 4 factors and 2 levels were A1B1CzD2 and A2BC2D2. SAS statistical analysis showed that at acute stage and adhesion stage, CZ Dz , meaning acupuncture with fifiform needling and warm needling therapy combined with positive functional exercise, is the main factor of the improvements of shoulder motor function (P<0.05).

CONCLUSIONFor periarthritis of shoulder at acute stage, the combined therapy of acupuncture at local acupoints, warm needling and positive functional exercise is adopted. At chronic stage, the combined therapy of acupuncture at local acupoints and distal acupoints, acupuncture with filiform needle and warm needling and positive functional exercise is the best program. Additionally, in clinical treatment, the patients' age, sex, shoulder joint function and duration of treatment should be considered comprehensively for the impacts on the efficacy.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Combined Modality Therapy ; Exercise Therapy ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Periarthritis ; therapy ; Shoulder Pain ; therapy

OBJECTIVETo investigate the cyto-genotoxicity of cigarette smoke condensates (CSCs) in human peripheral blood lymphocytes with different assays in vitro.

METHODSHuman lymphocytes were exposed to particle matter of cigarette smoke combined with or without S9 mixtures at doses of 25, 50, 75, 100 and 125 microg/ml for 3 h. The cytotoxicity induced by CSCs was detected by CCK-8 assay. The DNA damage, DNA repair (repair time: 30, 60, 90, 120 and 240 min, respectively) and the somatic cell mutations induced by 75 microg/ml CSCs were measured by comet assay, hprt gene and TCR gene mutation tests, respectively.

RESULTSCCK-8 assay indicated that the cell viability decreased with CSCs doses. At the doses of 100, 125 microg/ml, the cell viability of CSCs +S9 group was significantly higher than that of CSCs -S9 group (P < 0.05, P < 0.01). In comet assay, DNA damage significantly increased in a dose-dependent manner, as compared with controls (P < 0.01). Moreover, there was significant difference between -S9 group and +S9 group (P < 0.05, P < 0.01). The Mf-TCR at each dose group was significantly higher than that of controls (P < 0.05, P < 0.01). The Mf-hprt at high-dose groups were significantly higher than that of controls (P < 0.01), and significant difference of Mf-TCR and Mf-hprt at high doses of CSCs between -S9 group and +S9 group (P < 0.05, P < 0.01). The DNA damage induced by CSCs +S9 or CSCs -S9 could be repaired, but DNA repair speed was different between -S9 group and +S9 group (P < 0.05, P < 0.01).

CONCLUSIONCSCs may induce cyto-genotoxicity in human peripheral blood lymphocytes in vitro, but S9 mix could reduce the toxicity of CSCs and impact DNA repair speed.

Cells, Cultured ; Comet Assay ; DNA Damage ; drug effects ; DNA Repair ; drug effects ; Humans ; Lymphocytes ; drug effects ; Male ; Mutation ; Tobacco Smoke Pollution ; adverse effects ; Young Adult

OBJECTIVETo study the anti-platelet GPVI single chain Fv phage antibody which can inhibit the aggregation function of platelet by using phage antibody library technology.

METHODSITP patients with anti-platelet GPVI autoantibody that could inhibit the aggregation function of platelet were screened by MAIPA assay and platelet aggregation test. The gene fragments of heavy chain and light chain variable region (VH and VL) of immunoglobulin were amplified by RT-PCR from peripheral blood lymphocytes mRNA of the screened patients. The VH and and VL fragments were linked through a DNA linker encoding the peptide (Gly4Ser)3 to construct single chain Fv (ScFv) gene. The ScFv gene was digested with SfiI/NotI restriction enzymes and cloned into the pHEN2 phage display vector, then electrically transformed to E. coli TG1. The TG1 containing ScFv-pHEN2 was rescued by helper phage M13K07 to produce ScFv phage antibody. The anti-platelet GPVI phage ScFv antibody was enriched and purified. The effect of the phage antibody on platelet aggregation function was studied.

RESULTSOf 806 chronic ITP patients, 11 (1.36%) were positive for anti-platelet GPVI autoantibody and 2 (0.24%) patients'plasma significantly inhibited the collagen induced platelet aggregation. The length of VH and VL fragments was about 380 to 400 bp, and were successfully formed ScFv fragments of about 800 bp by DNA linker. After cloning ScFv to phagemid vector pHEN2 and transforming ScFv-pHEN2 to TG1, 4.1x10(7) clones were obtained. After M13K07 rescue, 2.62x10(10) cfu/ml ScFv phage antibodies were produced. The purified anti-platelet GPVI ScFv phage antibody inhibited the collagen induced platelet aggregation.

CONCLUSIONAnti-platelet GPVI ScFv phage antibody produced by phage antibody library technology can inhibit the aggregation function of platelet.

Adolescent ; Adult ; Aged ; Escherichia coli ; genetics ; Female ; Humans ; Male ; Middle Aged ; Peptide Library ; Platelet Aggregation ; immunology ; Platelet Aggregation Inhibitors ; immunology ; pharmacology ; Platelet Membrane Glycoproteins ; immunology ; Single-Chain Antibodies ; genetics ; immunology ; pharmacology ; Transformation, Bacterial ; Young Adult

OBJECTIVETo identify the factor XI gene mutation in a Chinese pedigree of congenital factor XI deficiency.

METHODSThe peripheral blood samples were collected from the proband and her family members and the plasma FXI:C and FXI:Ag were assayed. All the exons and their adjacent intron sequences of factor XI were amplified with PCR and sequenced thereafter.

RESULTSTwo novel nonsense mutations TGG-->TGA (Trp228stop) and TGG-->TAG (Trp383stop) were identified in the family.

CONCLUSIONThe compound heterozygous Trp228stop and Trp383stop may attribute to the pathogenesis of the congenital factor deficiency.

Adult ; Asian Continental Ancestry Group ; Codon, Nonsense ; Factor XI ; genetics ; Factor XI Deficiency ; congenital ; genetics ; Female ; Humans ; Male ; Middle Aged ; Pedigree ; Polymerase Chain Reaction ; Sequence Analysis, DNA

BACKGROUNDCoagulation factor VII (FVII) levels in plasma are usually related to ischemic heart disease (IHD) and cerebral infarction shares many of the risk factors related to IHD. Is there any relationship between factor VII and cerebral infarction? We investigated the relationship between FVII and acute cerebral infarction and reported genotype frequencies and allelic frequencies of FVII gene polymorphisms in the Chinese Han population.

METHODSWe recruited 62 patients with acute cerebral infarction confirmed by magnetic resonance imaging (MRI) from Ruijin Hospital, and 149 age-matched patients clinically free of vascular disease to act as controls. All of them were unrelated, and were from the Chinese Han population. FVII coagulant activity (FVIIc) was determined using an clotting assay, activated FVII (FVIIa) and FVII Ag were assayed using enzyme immunoassay kits. The FVII gene polymorphisms to be detected included-401G/T, -402G/A, 5'F7A1/A2, IVS7 and R353Q. 5'F7 and IVS7 were revealed by means of a PCR and direct agarose gel electrophoresis. The rest were examined by a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).

RESULTSThe results showed that FVIIc, FVIIAg and FVIIa were higher in the acute cerebral infarction group than in the control group (P < 0.01, P < 0.05, P < 0.05, respectively). There were no significant differences in the genotype frequencies of FVII gene polymorphisms between the two groups. The allelic frequencies in the Chinese Han population were as follows: -401G/T (96.64/3.36), -402G/A (52.01/47.99), 5'F7A1/A2 (96.64/3.36), IVS7 H5/H6/H7/H8 (0.34/52.35/46.98/0.34) and R353Q (95.64/4.36). There were significant differences (P < 0.01, P < 0.001, P < 0.001, P < 0.001, P < 0.001, respectively) in these allelic frequencies between the Chinese Han and European populations.

CONCLUSIONSThe results indicate that increased plasma FVII levels may contribute to thrombosis in cerebral infarction. And there was no significant difference in genotype frequencies of these five FVII gene polymorphisms between the acute cerebral infarction and control groups. Moreover, these results showed that the frequencies of protective allele, including -401T, 5'F7 A2 and 353Q were lower, but that -402A, which was previously found to be associated with increased plasma FVII levels, is higher in Chinese Han population.

Acute Disease ; Asian Continental Ancestry Group ; genetics ; Cerebral Infarction ; genetics ; China ; Europe ; European Continental Ancestry Group ; genetics ; Factor VII ; analysis ; genetics ; Gene Frequency ; Humans ; Intracranial Thrombosis ; genetics ; Polymorphism, Genetic

OBJECTIVETo explore the effect of autologous cytokine-induced killer cells on the quality of life in patient with breast cancer who have already finished the adjuvant chemotherapy.

METHODSOne hundred and twenty-eight postoperative patients with breast cancer who underwent anthracycline-based adjuvant chemotherapy were enrolled in this prospective study, and they were randomized into 2 groups, i.e., treatment group, which received the therapy of CIK cells transfusion, and control group, which was given regular follow-up. Meanwhile, patients with positive hormone receptor in the two groups were given endocrine therapy, and the patients with positive axillary lymph nodes were given radiotherapy to the chest wall and regional lymph nodes. The difference of quality of life between the two groups was analyzed according to the EORTC QLQ-BR53 quality of life questionnaire, and the adverse reactions were monitored.

RESULTSAs regarding the functional evaluation, the physical function scores of patients of the treatment group were (83.43 ± 14.87) and (88.55 ± 11.62) at 3 and 6 months after the CIK cell therapy, respectively, significantly higher than the baseline value [(74.83 ± 13.82), P < 0.05)]. Global health status/QOL scores were (83.30 ± 19.09) and (89.68 ± 10.81), significantly higher than the baseline value [(77.72 ± 21.05), P < 0.05]. As regarding symptoms, the scores of fatigue, nausea, vomiting and loss of appetite of patients in the treatment group were higher than the baseline value, with significant differences (P < 0.05). The nausea and vomiting scores in the control group at 3 and 6 months of followed-up were (26.67 ± 22.56) and (21.47 ± 21.06), significantly lower than the baseline values [(33.31 ± 27.07), P < 0.05]. The scores of worrying about the future in the patients of treatment group were (47.56 ± 30.84) and (42.33 ± 26.95) after 3 and 6 months, significantly better than the baseline value [(57.41 ± 30.63), P < 0.05]. The systematic therapy side effects scores were (31.95 ± 27.52) and (23.72 ± 22.87), significantly better than the baseline value [(40.56 ± 26.28), P < 0.05]. The scores of arm edema were (45.26 ± 25.42) and (36.61 ± 20.51), significantly milder than the baseline value [(55.11 ± 22.82), P < 0.05]. In the control group, the scores of arm edema were (44.85 ± 28.94) and (38.64 ± 23.68), significantly lower than the baseline values [(53.26 ± 23.84) points, P < 0.05]. Alopecia scores were (29.93 ± 24.72) and (24.18 ± 22.66), significantly lower than the baseline values [(35.92 ± 22.08), P < 0.05]. In the treatment group, the patients' physical function, social function and global health status/QOL, fatigue, insomnia, and worrying about the future rates were significantly higher than that of the control group (P < 0.05 for all). Three patients after CIK reinfusion had transient fever, and 6 cases felt pain in the lower limb, but the symptoms were relieved after symptomatic treatment.

CONCLUSIONSTherapy of autologous CIK cells transfusion can significantly improve the quality of life of breast cancer patients, and the adverse reactions during the treatment can be alleviated by symptomatic treatment.

Adult ; Anthracyclines ; administration & dosage ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast Neoplasms ; drug therapy ; surgery ; therapy ; Chemotherapy, Adjuvant ; Cytokine-Induced Killer Cells ; immunology ; transplantation ; Fatigue ; etiology ; Female ; Humans ; Immunotherapy, Adoptive ; adverse effects ; Middle Aged ; Nausea ; etiology ; Paclitaxel ; administration & dosage ; Prospective Studies ; Quality of Life ; Surveys and Questionnaires ; Vomiting ; etiology

BACKGROUNDAblation of complex fractionated atrial electrograms (CFAE) is an important adjunctive therapy in atrial fibrillation (AF). The present study was to elucidate the substrate underlying CFAE.

METHODSNine adult mongrel dogs were involved in the present study. AF was induced through rapid atrial pacing with vagosympathetic nerve stimulation. CFAE was recorded during AF. Ablation was performed at CFAE sites. Based on the location of the ablation scar, the atrial specimens were divided into CFAE and non-CFAE sites. Serial sections of the atrium were stained respectively with hematoxylin-eosin (HE) and the general neural marker protein gene product 9.5 (PGP9.5). We compared the characteristics of the myocardium and the ganglionated plexus (GPs) distribution between the CFAE and non-CFAE sites.

RESULTSThe myocardium of non-CFAE sites was well-organized with little intercellular substance. However, the myocardium in the CFAE site was disorganized with more interstitial tissue ((61.7 ± 24.3)% vs. (34.1 ± 9.2)%, P < 0.01). GPs in the CFAE site were more abundant than in non-CFAE sites ((34.45 ± 37.46) bundles/cm(2) vs. (6.73 ± 8.22) bundles/cm(2), P < 0.01).

CONCLUSIONThe heterogeneity of the myocardium and GPs distribution may account for the substrate of CFAE and serve as a potential target of ablation.

Animals ; Atrial Fibrillation ; pathology ; Dogs ; Electrophysiologic Techniques, Cardiac ; methods ; Myocardium ; pathology

OBJECTIVETo explore the gene mutation type of an inherited coagulation factor VII deficiency pedigree.

METHODSFVII:Ag, FVII:C, FVIIa were detected to classify deficiency type. FVII gene mutations were analysed in the proband and her family members by DNA directly sequencing. Biostructural pathology of the identified mutation was analysed by molecular modeling.

RESULTSHomozygosity of C-->T transition at position 11514 in exon 8 resulting in Thr359Met was identified in the proband, and heterozygosity for Thr359Met was confirmed in her parents, her son and some other family members. Thr359Met induces CRM-deficiency. It is found by computer simulated molecular model that the replacement of Thr by Met which has a larger and longer side chain might cause steric hindrance, and change the number of H-bonds.

CONCLUSIONSHomozygous missense mutation Thr359Met was found in a pedigree of hereditary FVII deficiency. This mutation might change the configuration of protein molecule and result in severe FVII deficiency.

Adolescent ; Adult ; Aged ; DNA Mutational Analysis ; Factor VII ; genetics ; Factor VII Deficiency ; genetics ; Female ; Homozygote ; Humans ; Male ; Middle Aged ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction

The expression of human clotting factor VIII gene was observed in transgenic off spring of mice through artificial insemination with sperm as carriers. Female mice were impregnated through artificial insemination by introducing sperm carrying pRC/RSV-hF VIII BD, which contained human F VIII BD (B-domain deleted) cDNA (hF VIII BD c DNA), into the uteri. During the fourth week after the birth of new-born mice, PCR was used to screen hF VIII BD cDNA positive transgenic mice, then blood of which was collected for detecting the antigen and Anti-hF VIII inhibitors, simultaneously, the transcription and expression of hF VIII BD cDNA were investigated by Northern blot and Western blot. The results showed that 7 became pregnant of 20 inseminated mice, and 11 new-born mice came into the world, out of which 9 survived at last. Three hF VIII BD cDNA-positive-transgenic mice had been screened out by PC R, in which the antigen of human F VIII in plasma was 8.65 ng/ml, 7.84 ng/ml and 8.44 ng/ml, respectively, the Anti-hF VIII inhibitors were all negative. Northern blot and Western blot showed that the transcription and expression of hF VIII BD cDNA existed in tissues such as spleen, liver, lung and kidney of 3 transgenic mice. It was concluded that transgenic mice carrying human F VIII gene can be generated by sperm-carrier techniques and express human F VIII protein. This experiment provides important data for manufacturing transgenic animal carrying human F VIII gene, which can work as a biological reactor to produce human F VIII protein, through sperm-carrier techniques.
Animals ; Blotting, Northern ; Blotting, Western ; Factor VIII ; genetics ; Female ; Gene Expression ; Humans ; Insemination, Artificial ; Male ; Mice ; Mice, Transgenic ; Pregnancy ; Spermatozoa ; metabolism

Rabdosia japonica(Burm.f.) Hara var.glaucocalyx(Maxim.) Hara is a traditional Chinese medicine, and is known to have anti-tumor effects. This study aims to investigate the effect of glaucocalyxin A (GLA), a diterpenoids extracted from Glaucocalyx Hara, on apoptosis of glioma cells and its mechanism. This study investigated the molecular signaling mechanism of GLA-induced glioma cell apoptosis by analyzing survival rate of C6 rat glioma cells, cell morphology, colony formation ability, interference ribonucleic acid, polymerase chain reaction, and Western blot. The result showed that in the presentce of GLA, the survival rate of C6 rat glioma cells decreased significantly, while the expression of guanine nucleotide-exchange factor-H1 was up-regulated, causing phosphorylation of extracellular regulated protein kinases proteins and apoptosis. Hence, the mechanism of GLA-induced glioma cell apoptosis was the GEF-H1/ERK pathway.