AIM: To investigate the amiloride-sensitive Na~+ channel current (Iamil) in primary cultured human bronchial epithelium cells of chronic obstructive pulmonary disease (COPD) patients and the influence of interleukin 4 and interleukin 6. METHODS: Human bronchial epithelium cells were isolated and cultured from 18 patients undergone pueumonectomy in hospital. Interleukin 4 or interleukin 6 at concentration of 10 ?g/L was used to treat these cultured cells, and Iamil was measured by whole cell patch clamp techniques. RESULTS: There was no markedly difference among normal no-smoking group, normal smoking group and COPD group. Interleukin 4 down-regulated the Iamil in normal no-smoking group, normal smoking group and COPD group. The down-regulated percentages were 59.7%, 54.7% and 30.0%. Interleukin 4 down-regulated the Iamil in normal no-smoking group (44.8%) and normal smoking group (34.9%) but not in COPD group, and the current forms were not changed after IL-4 or IL-6 treatment. CONCLUSIONS: Interleukin 4 and interleukin 6 down-regulated the Iamil in primary cultured human bronchial epithelium cells. It may contribute to the hypersecretion of COPD, and the up-regulated interleukin 4 and interleukin 6 in COPD patients may cause them react weaker to the treatment of interleukin 4 and interleukin 6.

ObjectiveTo observe the effects of hypoxia inducible factor-1 alpha (HIF-1α)-targeting small interfering RNA(siRNA) on the expression of HIF-1α and vascular endothelial growth factor (VEGF) in HaCaT ceils under hypoxic conditions. MethodsHaCaT cells were cultured and divided into four groups, normal control group (without any treatment), hypoxia group (cultured under hypoxic conditions for 24 hours),liposome control group (transfected with liposome followed by hypoxic culture for 24 hours), RNA interference group (transfected with HIF-1α-targeting siRNA/liposome complexes followed by hypoxic culture for 24 hours). Fluorescence real-time quantitative PCR was utilized to determine HIF-1oα and VEGF mRNA expression in HaCaT cells, and Western blot to detect HIF-1α and VEGF protein expression. ResultsNo significant difference was observed in the mRNA expression of HIF-1α between the hypoxia group and normal control group(0.907 ± 0.032 vs. 0.878 ± 0.034, F =1.108, P ＞ 0.05), while the expression levels of VEGF mRNA,HIF-1α and VEGF proteins were significantly higher in the hypoxia group than in the normal control group (0.935 ± 0.032 vs. 0.652 ± 0.053, 0.813 ± 0.047 vs. 0.236 ± 0.014, 0.791 ± 0.030 vs. 0.316 ± 0.013, all P ＜0.05). A significant decline was noted in the mRNA and protein expression levels of VEGF (0.230 ± 0.044 vs.0.978 ± 0.030, 0.213 ± 0.026 vs. 0.817 ± 0.049, both P ＜ 0.05) and HIF-1α(0.497 ± 0.033 vs. 0.806 ±0.040, 0.249 ± 0.028 vs. 0.833 ± 0.052, both P ＜ 0.05) in the RNA interference group than in the liposome control group. ConclusionsHypoxia may enhance the expression of HIF-1α and VEGF in HaCaT cells, and to inhibit the HIF-1α expression may suppress the expression of VEGF in HaCaT cells under hypoxia.

Objective To investigate the clinical efficacy of scalp acupuncture plus electroacupuncture in treating migraine without aura.Methods Three hundred and twenty-six patients with migraine without aura were randomly allocated to a treatment group of 157 cases and a control group of 169 cases. The treatment group received scalp acupuncture plus electroacupuncture and the control group, medication alone. The Pain Rating Index (PRI) score and the TCM syndrome rating scale score were scored in the two groups of patients before and after treatment. The numbers of taking medicine and the numbers of headache attacks were compared between the two groups of patients during treatment. An analysis was made of factors influencing the therapeutic effect on migraine. Results There were statistically significant differences in the PRI score and the TCM syndrome rating scale score at the end of treatment and at one, two and three months of follow-up compared with before treatment in the two groups (P<0.05). There were statistically significant differences in the PRI score and the TCM syndrome rating scale score at the end of treatment and at one and two months after it between the treatment and control groups (P<0.05). There was a statistically significant difference in the weekly number of taking medicine between the treatment and control groups during treatment (P<0.05). There was a statistically significant difference in the number of headache attacks between the treatment and control groups at two, three and four weeks of treatment (P<0.05). In the treatment group, the therapeutic effect at one, two and three months of follow-up was related to both TCM syndrome type and the VAS score; the therapeutic effect at two months of follow-up was related to the sex; the therapeutic effect at three months of follow-up was related to the educational level.Conclusion Scalp acupuncture plus electroacupuncture is an effective way to treat migraine without aura.

To examine the mRNA expression of matrix metalloproteinase 9 (MMP-9) and the gelatinase activity of its inhibitor, tissue inhibitor of matrix metalloproteinase 1 (TIMP-1) in the primary epithelial cells of patients with COPD, airway epithelial cells were taken from 15 COPD patients and cultured in vitro. The patients were divided into three groups, COPD group, normal smoking control group and non-smoking control group, with 5 subjects in each group, on basis of the smoking history and lung function. The semi-qualitative RT-PCR was employed to determine the mRNA levels of MMP 9 and TIMP-1 and SDS PAGE was used for the determination of the gelatinase activity of MMP-9 and TIMP-1. Our result showed that the mRNA of MMP-9 and TIMP-1 in epithelial cells of the non-smoking subjects was at a low level The mRNA of MMP 9 and TIMP-1 in COPD patients and smokers was significantly higher than that in non-smokers (P＜0.05). No significant difference was found in the levels of MMP-9 and TIMP-1 in epithelial cells between the COPD patients and smokers. The MMP-9/TIMP-1 ratios in COPD patients and smokers were significantly lower than that of non-smokers (P＜0.05). The gelatinase activity in the epithelial cells of both COPD patients and normal smokers was increased (P＜0.05), but no difference existed in the gelatinase activity in the epithelial cells between COPD patients and normal smokers. It is concluded that the transcription of MMP-9 and TIMP-1 and the gelatinase activity of MMP-9 and MMP-2 in the epithelial cells in COPD patients were increased, which resulted in an imbalance of MMP-9/TIMP-1, thereby causing pulmonary fibrosis. These factors play important roles in the pathogenesis of COPD.

OBJECTIVE: To investigate the expression of inducible nitric oxide synthase (iNOS) and hypoxia-inducible factor 1(HIF-1) α in psoriatic lesions,and to explore their relationship with angiogenesis in psoriasis. METHODS: HIF-1α and iNOS protein were detected by immunohistochemistry and Western blot in 32 cases of psoriasis and 20 healthy controls,and CD34 marking vascular endothelial cells were used to measure the microvascular density (MVD). RESULTS: The expressions of HIF-1α and iNOS protein were very weak in the control skin, but very strong in psoriatic lesions, which showed significant difference in the expressions of iNOS and HIF-1α between the psoriasis and the control group(P<0.05). Expression of HIF-1α (r=0.743, P<0.01) and iNOS (r=0.639,P<0.01) had positive correlation with MVD in psoriasis respectively. There was a positive correlation between iNOS and HIF-1α expression in psoriasis (r=0.717, P<0.01). CONCLUSION Both iNOS and HIF-1α have high expression in psoriasis and might play an important role in the genesis and development of psoriasis.
Adolescent ; Adult ; Biopsy ; Child ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Male ; Middle Aged ; Neovascularization, Pathologic ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Psoriasis ; metabolism ; pathology ; Skin ; blood supply ; metabolism ; pathology ; Young Adult

OBJECTIVE: To observe the effect of hypoxia inducible factor -1α (HIF-1α) small interfering RNA (siRNA) on the expression of HIF-1α and inducible nitric oxide synthase (iNOS) in HaCaT cells under hypoxia. METHODS: HaCaT cells were divided into 4 groups: the normal control group (without any treatment), the hypoxia group (under hypoxia for 24 h), the liposome control group (HaCaT cells transfected with liposome before hypoxia treatment), the RNA interference group (HaCaT cells transfected with siRNA sequences then under hypoxia for 24 h). Real-time PCR and Western blot were utilized to determine HIF-1α and iNOS mRNA and protein expression in HaCaT cells. RESULTS: There was no significant difference of the mRNA expression of HIF-1α between the hypoxia group and the normoxia group (P>0.05), but the protein expressions of HIF-1α was increased in the hypoxic group than that in the normoxia group (P<0.05). Both the mRNA and protein expression of iNOS were increased in hypoxic conditions than that in the normoxia (P<0.05). Decreases were more significant in the mRNA and protein expression of HIF-1α and iNOS in the RNA interference group than that in the liposome control group in HaCaT cells (P<0.05). CONCLUSION Hypoxia increased HIF-1α and iNOS expression in HaCaT cells and inhibition of HIF-1α expression decreased iNOS expression.
Cell Hypoxia ; Cell Line ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Keratinocytes ; cytology ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics

【Objective】 A framework to support nurses and midwives making the clinical decision and providing the written instruction for blood transfusion has been developed and implemented in the United Kingdom as a response to the changing needs of the patient and in recognition that blood transfusion services to patients could be improved by using the untapped knowledge and expertise of experienced nurses and midwives.Special education and training program for this role development are provided jointly by the national blood and nurse management authority, higher education institutions and transfusion societies.The British government has issued and implemented a compulsory professional indemnity which cover nurses and midwives as well.The development and implementation of the framework, policies and procedures for this role development is based on the regulatory compliance and the collaboration of, and beneficial to the multiple stakeholders, with the gaps left by doctors being fillled, work load of doctors reduced, nurses and midwives achieving professional development, hospitals performing more efficiently, and most importantly, the patients having a better transfusion services.At present, there is no similar policy or program for nurses and midwives in China.Therefore, this paper introduces the policy framework and implementation for this role development in UK, which would be a valuable reference for the role development and extension of nurses and the organization, education and training for transfusion professional teams as well in China in the near future.

OBJECTIVETo identify an inbred Chinese pedigree with autosomal recessive muscular dystrophy and analyze the molecular defects.

METHODSLinkage analysis was conducted using short tandem repeat(STR) markers from the regions associated with limb-girdle muscular dystrophy type 2A(LGMD2A) through 2H. Multi-Western blot was performed with anti-calpain-3, anti-dysferlin, anti-gamma-sarcoglycan, anti-alpha-sarcoglycan, and anti-dystrophin monoclonal antibodies. Mutation was determined by reverse transcriptase-polymerase chain reaction and sequencing.

RESULTSTwo-point linkage analysis showed significant Lod scores with markers from chromosome 2p13, the highest two-point Lod scores were obtained with D2S337 (Z(max)=1.86 at theta=0). Multi-Western blot confirmed dysferlin deficiency of muscle specimen from the proband. Mutation analysis revealed a novel 6429delG mutation on exon 53 of the DYSF gene for the proband.

CONCLUSIONThe authors identified an inbred Chinese pedigree with Miyoshi myopathy caused by a 6429delG on the DYSF gene. This mutation is predicted to result in premature termination of translation.

DNA, Complementary ; chemistry ; Dysferlin ; Genetic Linkage ; Humans ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Muscle Proteins ; genetics ; Muscular Diseases ; genetics ; Muscular Dystrophies ; genetics ; Mutation ; Pedigree