Method of isolation and purification of recombinant hepcidin was described, and the bioactivity of the protein was assayed in this paper.The oxidation of his-hepcidin was carried out in the cysteine-cystine system, and the multimers were removed through gel filtration under denaturation condition.Then the protein was refolded by continuous dilution and digested by enterokinase.The total yield of his-hepcidin before enterokinase cleavage is 50%, and the purity is above 95%.Through agar diffusion assay, the recombinant hepcidin displayed obvious antibacterial activity against B.subtilis.The LC-ESI-MS analysis of recombinant hepcidin showed that the measured molecular weight accorded with the calculated molecular weight, and the CD spectrum indicated that the secondary structure of recombinant hepcidin is similar with native hepcidin.

Data Collection ; methods ; Metabolomics ; methods

Objective To discuss the Clinical character of pulmonary tuberculosis combined malignant lymphoma and its Pathogenesis,and to review the literature.Methods Eighteen cases of pulmonary tuberculosis combined malignant lymphoma from 1996 to 2003 were retrospectively analyzed by its clinical manifestations,X-ray features,diagnosis and treatments.Results 18 cases were all infiltrative pulmonary tuberculosis,13 them were calcification focals,5 were active pulmonary tuberculosis;5 of all were Hodgkin’s lymphomas,13 of all were non-Hodgkin′s lymphomas.16 cases were lymphomas after tuberculosis,2 cases tuberculosis after lymphomas,none were co-existent malignant lymphoma and tuberculosis.Tuberculosis may precede or complicate a lymphomatous process during the development of both diseases,This might is linked to immune deficiency and chronic inflammation;Lymphomas might cause pulmonary tuberculosis,it might cause the immune turbulence of an individual,Pulmonary tuberculosis infection occurring during or after the radiotherapy and chemotherapy of lymphoma.Conclusions It may pulmonary tuberculosis combined malignant lymphoma in the patients in the endemic areas of tuberculosis,Appropriate invasive biopsy procedures are necessary for early diagnosis.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Hepatitis B ; epidemiology ; immunology ; prevention & control ; Humans ; Infant ; Infant, Newborn ; Male

Objective To study the relationship between self-efficacy and self-management behaviors in convalescent patients with acute pancreatitis.Methods Totals of 70 convalescent patients with acute pancreatitis were investegeted with self-efficacy scale and self-management behaviors questionnaire,then collected score respectively,analyzed the relation between self-efficacy and self-management behaviors.Results The score of selfmanagement questionnaire was( 14.91 ± 3.21 ),85.7％ patients had a high management level,and only 40％patients of them had a high level of exercise,34.2％ patients had a low level of bad living life management.The score of self-efficacy scale was( 15.23 ±3.53),and 37.1％ patients had a high self-efficacy level,48.6％ patients had a higher level,14.3％ patients had a medium level,no low self-efficacy was found.Self-efficacy in convalescent patients with acute pancreatitis was positively correlated with self-management and self-management level( r =0.803,P ＜ 0.01 ).Kinds of self-efficacy were positively correlated with related self-management ( r =0.235 -0.843,P ＜ 0.05 ).Conclusions Convalescent patients with acute pancreatitis need to enhance exercises and health education about bad living life management and scientifically use self-efficacy theory,se as to reduce disease recurrence and improve patients' life quality.

OBJECTIVETo explore the effect of Bushen Gujin Recipe (BGR) on serum and synovial expression of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-α) in knee osteoarthritis (KOA) model rabbits.

METHODSTotally 36 8-month-old healthy male New Zealand white rabbits were randomly divided into 4 groups, i.e., the normal control group, the model group, the Western medicine group (Meloxicam, at the daily dose of 6 mg/kg), and the TCM group (BGR, at the daily dose of 53 g/kg), 9 in each group. Modeling was performed in all rabbits except those in the normal control group by using Hulth A method. All medication was performed for 8 consecutive weeks. Contents of IL-1 and TNF-α were detected using ELISA from serum, partial synovial tissue of the front knee joint, cartilage and subchondral bone of the medial femoral condyle.

RESULTSThe joint space became narrowed in the Western medicine group, ranging between the model group and the TCM group. The articular surface was rough with obvious osteophytes. The joint space was slightly narrower in the TCM group; the articular surface was slightly rough with mild osteophytes. Compared with the normal control group, contents of IL-1 and TNF-α in serum and synovial increased in the model group (P < 0.01). Compared with the model group, contents of IL-1 and TNF-α in serum and the synovial fluid decreased in the two treatment groups (P < 0.01). There was no statistical difference in contents of IL-1 and TNF-α between the Western medicine group and the TCM group.

CONCLUSIONBGR promoted the synthesis of cartilage matrix and carti- lage repair through inhibiting the secretion of IL-1 and TNF-α, and prolonging cartilage degeneration.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Interleukin-1 ; metabolism ; Knee Joint ; Male ; Osteoarthritis, Knee ; drug therapy ; metabolism ; Rabbits ; Synovial Fluid ; Synovial Membrane ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective: To observe the alteration of type-　 Ⅰ collagen protein gene expression after arterial injury and investigate its effect on the development of restenosis. Methods: Firstly, thee xperimental carotid arterial injury rabbit model was constructed. Then, Norther n blot, in situ hybridization and histomorphometric analysis were used to de tect the expression of procollagen mRNA and the accumulation of collagen protein 1,2,4 weeks after injury. Results: Type-　Ⅰ collag en mRNA increased 1 week after injury, peaked 2 weeks later and decreased 4 week s later. The deposition of the collagen protein account for a high percentage o f space in neointima on histomorphometric analysis. Conclusion: Collagen protein may play an important role in the development of neointima and restenosis.

Objective: To observe the clinical effects of moxibustion at abdominal acupoints for slow transit constipation (STC) due to yang deficiency of the spleen and kidney. Methods: A total of 52 cases with slow transit constipation in conformity with the inclusion criteria were selected and divided into a control group and an observation group according to their visit order and random digital table, 26 cases in each group. Patients in the control group received routine nursing guide. Besides the same routine nursing guide, patients in the observation group received moxibustion at the abdominal acupoints, once every day. The course of the treatment was 4 weeks in the two groups, and the 3-month follow-up was given after the course was finished, for comparing the clinical symptoms, results of colon transit tests, scores of depression/anxiety scale and nursing satisfaction. Results: The total effective rate was 92.3% in the observation group and 69.2% in the control group, with a significant difference between the two groups. After the treatment and during the follow-up checks, the scores of Chinese medical symptoms in the two groups were remarkably decreased than those before the treatment (all P<0.01); the scores of the observation group were obviously lower than those in the control group (all P<0.01). The discharge rates of the markers in the two groups were remarkably increased than those at the same time period before the treatment; moreover, the discharge rates of the markers at various time periods were remarkably better in the observation group than those in the control group (P<0.01). SDS and SAS scores were remarkably decreased after the treatment in the two groups (P<0.01). In comparison of SDS score between the two groups after the treatment, the difference was statistically significant (P<0.01). In comparison of SAS score between the two groups after the treatment, the difference was statistically significant (P<0.05). The nursing satisfaction was 96.2% in the observation group after the treatment, obviously better than that in the control group (73.1%). The recurrence rate was 8.3% in the observation group, remarkably lower than that in the control group (33.3%). Conclusion: Moxibustion at the abdominal acupoints plus routine nursing can remarkably improve the colon transit functions and anxious and depressive emotion in patients with STC, and the therapeutic effects are remarkable. Not only the clinical satisfaction is higher, but the recurrence rate is obviously lower than that of routine nursing.

Objective To determine the difference of macrophage RAW264.7 apoptosis induced by Brucella melitensis virulent strain 16M and attenuated strain M5-90 and elucidate the regulatory role of caspases 3,8 and 9.Methods The best multiplicity of infection (MOI) was determined through kinetic analysis of Brucella melitensis strain 16M and M5-90 induced mouse macrophages apoptosis(bacterium ∶ cell =100 ∶ 1,50 ∶ 1,10 ∶1).The infection model was established using the best MOI =50 ∶ 1.The numbers of in vivo bacteria by colony formation units were calculated after macrophages were infected for different times,including 2,4,8,12,24 and 48 h,and the infected cells were collected.The ratios of apoptosis were detected and the regulation of caspases 3,8 and 9 in apoptosis pathway was elucidated by flow cytometry.Results The numbers of 16M in vivo bacteria were 105.4,104.8,105.8,106.5,108.0 and 109.0,respectively and of M5-90 were 106.1,106.2,106.4,106.3,106.1 and 105.0,respectively.The number of in vivo bacteria of 16M was significantly increased than that of M5-90 after infected for 24 h to 48 h.The ratios of apoptosis induced by 16M after infected for 2,4,8,12,24 and 48 h was (2.67 ± 0.09)％,(13.13 ± 0.30)％,(6.56 ± 0.42)％,(6.49 ± 0.28)％,(16.07 ± 0.86)％ and (24.23 ± 1.67)％,respectively,and by M5-90 was (3.62 ± 0.02)％,(32.01 ± 2.59)％,(17.58 ± 0.44)％,(16.09 ± 0.10)％,(62.53 ± 2.70)％ and (85.53 ± 0.15)％,respectively,and by control group was [(1.90 ± 0.20)％,(1.92 ±0.16)％,(1.99 ± 0.03)％,(2.48 ± 0.11)％,(3.56 ± 0.07)％,(5.26 ± 0.33)％].The differences were statistically between groups in same time.The Brucella melitensis vaccine strain M5-90 was more powerful than virulent strain 16M in respect of inducing macrophage apoptosis after infected for 24 to 48 h.Twenty-four hours after infection,the expression of caspases 3,8 and 9 was (1.47 ± 0.05)％,(1.52 ± 0.02)％ and (2.47 ± 0.12)％,respectively,in control group and the expression was (9.70 ± 0.46)％,(6.08 ± 0.56)％ and (35.08 ± 1.64)％,respectively,after infected for 24 h induced by M5-90.The expression of caspases 3,8 and 9 was significantly higher than that control group (P ＜ 0.01).Twenty-four hours after given caspases 3,8 and 9 inhibitor,apoptosis rate in control group was (66.72 ± 1.28)％,in M5-90 group was (22.58 ± 0.55)％,(53.15 ± 1.85)％ and (29.18 ± 0.23)％,respectively,and compared with control group,apoptosis rate of caspases 3,8 and 9 was significantly lower(P ＜ 0.01).Conclusions Apoptosis of macrophage can be induced by Brucella melitensis virulent vaccine strain 16M and attenuated strain M5-90.M5-90 is stronger than that of strain 16M.Caspases 3,8 and 9 can regulate macrophage apoptosis after M5-90 infection.

OBJECTIVETo study the mechanism of Bushen Qiangdu Recipe (BQR) for regulating osteoprotegerin receptor activator for nuclear factor kappa B ligand (OPG/RANKL) pathway in ankylosing spondylitis (AS).

METHODSThirty active AS inpatients or outpatients were recruited from Department of CM Rheumatology, China-Japan Friendship Hospital from January to May 2009. All patients were treated with BQR for 3 successive months, one dose daily, once in the morning and once in the evening. Besides, 30 healthy volunteers were recruited. The serum of patients and volunteers were collected. The osteoblast cell lines hFOB1. 19 were divided into 3 groups: the pre-treatment group, the post-treatment group, and the healthy volunteer group (as the control group). All cell lines were cultured by corresponding culture medium containing each serum. The supernatant from osteoblast cell lines was collected. The protein content of OPG/RANKL was detected using ELISA, and the protein expression of OPG/RANKL was detected using RT-PCR.

RESULTSCompared with the control group, the OPG content, the mRNA and protein expressions of OPG, and the mRNA and protein expressions of OPG/RANKL all decreased, while the mRNA expression of RANKL increased in the pre-treatment group, showing statistical difference (P<0.05, P<0.01). Compared with the pre-treatment group, the OPG content, the mRNA and protein expressions of OPG significantly increased, and the mRNA and protein expressions of OPG/RANKL increased, while the mRNA expression of RANKL decreased in the post-treatment group, showing statistical difference (P<0.05, P<0.01).

CONCLUSIONSAS patients' serum could directly inhibit the expression of OPG in osteoblasts, promote the expression of RANKL, and down-regulate the OPG/RANKL ratio. BQR containing serum might promote the osteogenesis and inhibit the bone resorption possibly through directly up-regulating the OPG/RANKL ratio in osteoblast, thus inhibiting the differentiation and function of osteoclast.

Adult ; Case-Control Studies ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Male ; Osteoblasts ; drug effects ; metabolism ; Osteogenesis ; Osteoprotegerin ; metabolism ; RANK Ligand ; metabolism ; Serum ; Spondylitis, Ankylosing ; metabolism ; Young Adult