Objective:To obtain a large amount of erythromycin B and to investigate the activity site in eryK. Methods:The key sequence of the BC loop region in eryK gene was knocked out and the eryK gene with 101 bp deleted was amplified by overlapping PCR,and cloned into vector pWHM3 to construct recombinant plasmid. The Saccharopolyspora erythraea mutant AK17 was constructed through chromosomal homologous recombination technique.Results and Conclusions:The S.erythraea mutant AK17 was constructed. The results of TCL and MS analysis showed that the major fermentation product of AK17 is erythromycin B.

Objective To investigate the proliferative characteristics of fibroblast-like synovial cells (FLS)in osteoarthritis in vitro and the mechanism of the immunosuppressive effect of total glucusides of paeony(TGP).Methods FLS of OA and non-inflamed synovium(NS)were cultured and identified in vitro in the presence or absence of TGP.After incubation,the survival fraction(SF)of FLS was evaluated by MTI' and the TNF-?,IFN-?and bFGF level in cultured FLS supernatant was measured by ELISA.The expression of FLS c-los mRNA and cell cycle of OA-FLS was observed by RT-PCR and flow eytometry respectively at the same time.Results No statistical significant differences were noted between the OA and NS FLS in pro- liferating double time.High doses of TGP suppressed FLS-SF more evidently in OA patients than in NS(P0.05).Conclusion High dose TGP can inhibit OA-FLS proliferation,modulate cy- tokine secretion and c-fos expression in OA.This suggests that TGP has immunosuppressive effect on OA syn- ovitis,probably by preventing the synovial hypertrophy in OA.

Objective To analyze the epidemiological characteristics of Mycobacterium tuberculosis by enterbaeterial repetitive intergenic consensus-polymerase chain reaction(ERIC-PCR)DNA fingerprint. Methods Mycobacterium tuberculosis positive sputum samples between September 2003 to May 2006 were collected and cultured.Chromosomal DNA were extracted and ERIC-PCR DNA fingerprinting was analyzed by software,such as RAPD PHYLIP and Treeview.Results A total of 42 different fingerprints were detected.Phylogenetic analysis showed that they could be classified into three clusters,the clustering rate was 72.6%.The characteristics of ERIC-PCR fingerprint patterns were related to age,drug resistance,and type of resistance.Conclusions ERIC-PCR DNA fingerprinting technique used in this study is good for epidemiological studies with its strong discrimination,simplicity and rapidness.A high level of recent transmission is found in our city.

Objective To develop a convenient method efficiently expands the frequency of specific CTLS.Methods We used different concentrations of CMV-speeific epitope peptides pp65 to stimulate PBMCs for expansion of CMV-specific CTLs.CMV-specifie CTLs were doubly labeled by tetramers-PE and CD_8-FITC for FACS analysis.Results The method expands CMV-speeific CTLs efficiently.CMV-specific CTLs were expanded from 1% to 20% of PBMCs quickly(namely 40% of CD_8~+ T cells).The method provided a large number of cells with tetramer staining of CD_8~+ T cells for FACS analysis from a single blood sampling.Conclusions Peptides stimulation methods are convenient,easy to operate and expanded CMV- specific CTLs efficiently.The increased frequencies of CMV-specific CTLs allowed the data of different individuals to be easily compared and sequentially evaluated.The methods lay the base for adoptive immunotherapy to prevent CMV disease.

In this study we present our experiences with the reverse sural fasciocutaneous flap to reconstruct the distal lower limb soft tissue defects caused by traumatic injuries. These flap graftings were carried out from Oct. 2010 to Dec. 2012 in our department. The series consisted of 36 patients, including 21 men and 15 women with an average age of 46.2 years (14-83 years) and with a medium follow-up period of 18 months (12-24 months). Of all the cases of acute trauma, there were 10 cases of trauma of distal tibia, 9 cases of trauma of perimalleolus, and 17 cases of trauma of midfoot and forefoot. Related risk factors in the patients were diabetes (2 cases), advanced age (>65 years, 3 cases) and cigarette smoking (6 cases). The reverse flow sural island flap irrigation depended on lower perforators of the peroneal artery. The fasciocutaneous pedicle was 3-4 cm in width and the anatomical structures consisted of the superficial and deep fascia, the sural nerve, short saphenous vein, superficial sural artery together with an islet of subcutaneous cellular tissue and skin. The most proximal border of the flap was only 1.5 cm away from the popliteal skin crease and the pivot point was 5-7 cm above the tip of the lateral malleolus. All the flaps survived. No arterial crisis occurred in any case. The venous congestion occurred in 2 cases and got better after raising the limbs and bloodletting. Only in an old man, 1.5 cm necrosis of distal margin of his flap occurred and finally healed after continuous dressing change. One-stage skin grafting was performed, and all the donor sites were sutured and successfully healed. It was concluded that the reverse sural fasciocutaneous flap is safe and reliable to extend to the proximal third even near the popliteal skin crease. We also concluded this flap can be safely and efficiently used to treat patients with large and far soft-tissue defects from the distal leg to the forefoot with more versatility and it is easier to reach the recipient sites.

Objective To investigate the effects of partial body weight supported treadmill training (BW- STT) on post-stroke depression (PSD) and on patients' quality of life.Methods Sixty patients with PSD were re- cruited and divided into a training group (n=30,male 17,female 13) and a control group (n=30,male 16,fe- male 14).All patients were treated with routine internal medication and rehabilitation.The patients of the training group also received BWSTT in addition to their routine treatment.All patients' neurological impairment was evaluated using the Modified Edinburgh-Scandinavian Stroke Scale (MESSS).The Hamilton depression scale (HAMD) was used for evaluating the degree of depression.The Fugl-Meyer scale and the Barthel index were used to assess ambula- tion and balance,and facility in the activities of daily living.All patients were assessed before and after the treat- ment.Results After four weeks of treatment,depression in the training group had improved significantly more than in the control group.Conclusion BWSTT intervention is very important for patients with PSD:it can reduce the degree of depression and improve the quality of life.

Objective To investigate the proliferation characteristics of fibroblast like synovial cells (FLS)in osteoarthritis in vitro and the mechanism of the immnnosuppressive effect of T-614 [N-(3-formy- lamino-4-oxo-6-phenoxy-4H-chromen-7-yl)methanesulfonamide ] on them.Methods FLS of OA and non- inflamed synovium(NS)were cultured and identified in vitro in the presence or absence of T-614.After incu- bation,the survival fraction(SF)of FLS was evaluated by MTT,cell cycle was observed using fluorescence - activated cell sorting(FCS)method and the expression of c-fos and COX-2 mRNA was examined by RT- PCR in FLS of OA patients.Results No statistically significant difference was noted between the OA and NS FLS in proliferation ability and cell cycle.High dose T-614 suppressed FLS SF obviously in OA and NS sta- tistically(P＜0.05),whereas the inhibition degree was not different between the two kinds of FLS.The agent induced cell apoptosis and reduced the accumulation of c-fos mRNA in OA-FLS at dose 1000 ml/L,prolonged G_1 term and shortened S term at dose 200 ml/L.The expression of COX-2 mRNA in OA FLS was suppressed obviously by T-614 at dose 1000 ml/L.Conclusion OA FLS do not display a distinct activated unlimited viability compared with NS cells,without stimulated by proinflammatory cytokine in vitro.High dose T-614 moderately inhibits the proliferation and differentiation of FLS,directly affects gene of the c-fos and COX-2 expression in OA,which may contribute to its immunosuppressive effect on OA'synovitis.

Objective To investigate the proliferation and differentiation characteristics of fibroblast like synovial cells(FLS)in rheumatoid arthritis(RA)in vitro and the mechanism of the immunosuppressive effect of differentiation inducers, such as all trans retinoid acid(ATRA), ealcitriol [1,25(OH)_2D_3] and dexamethasone(DEX). Methods FLS of knee synovial tissues from RA patients were cultured and identified in vitro in the presence or absence of ATRA, 1,25(OH)_2D_3 and DEX respectively. Synoviocyte proliferation in RA were measured by MTT colorimetrie assay and the survival fraction(SF)of FLS was evaluated. Cell cycle of FLS was observed using fluorescence-activated cell sorting(FCS)method in RA patients. Results The identified synovial cells in patients with RA were FLS(Vimentin and Fibronectin expression was positive), and hadn't been transformed or differentiated to adipocytes and osteoblasts with the three inducers. The SF of all RA-FLS interfered by ATRA, 1,25(OH)_2D_3 and DEX was much lower than that without drugs vehicle group in RA-FLS(P

OBJECTIVETo observe the clinical effect in treatment of the recurrent allergic (Henoch-Schönlein) purpura and prevention of renal impairment by means of integrative Chinese and Western medicine (ICWM).

METHODSOne hundred and seventy-six children with allergic purpura were randomly divided into the treated group (Based on the Western routine therapy of the control group, Kangmin Xiaoban Yin, a kind of TCM decoction, was added for 7-10 days) and the control group. Levels of urinary beta2-microglobulin (beta2-MG) and serum IgA, IgE were measured before and after treatment. And clinical observation on the disappearance time (DT) of abdominal pain, arthralgia and purpura were implemented.

RESULTSThe DT of joint swelling, arthralgia, and purpura in the treated group was shorter than those in the control group, showing significant difference (P<0.01). Comparison between groups in urinary beta2-MG and serum IgA and IgE before treatment and 1 week after treatment were insignificantly different (P>0.05), but at 1 month and 6 months after treatment, the level of urinary beta2-MG in the treated group was lower than that in the control group, with significant difference (P<0.05), and levels of serum IgA and IgE at 1 month after treatment in the former were lower than those in the control group, also showing significant difference (P<0.05, P<0.01). Six-month recurreat rate in the treated group was 23.5%, while that in the control group. There was significant difference in comparison in 1/2-year recurrent rate (P<0.01).

CONCLUSIONICWM in treating allergic purpura could reduce the repeated episodes and shorten the hospitalization time, and effectively prevent the renal impairment of the disease.

Adolescent ; Child ; Child, Preschool ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunoglobulin A ; blood ; Immunoglobulin E ; blood ; Kidney Diseases ; etiology ; prevention & control ; Male ; Phytotherapy ; Prednisone ; therapeutic use ; Purpura, Schoenlein-Henoch ; complications ; drug therapy ; Secondary Prevention ; beta 2-Microglobulin ; urine

OBJECTIVETo investigate whether myocardial bridging (MB) is independently associated with coronary atherosclerosis proximal to MB in the left anterior descending coronary artery (LAD) identified by computed tomographic coronary angiography (CCTA).

METHODSFrom March 2011 to December 2012, patients (n=9 862) with suspected coronary disease underwent CCTA using dual-source CT scanner. The baseline clinical characteristics (age, gender, smoking history, presence of hypertension, dyslipidemia, diabetes mellitus, family history of heart attack, and body mass index) and the results of CCTA were reviewed. Two radiologists evaluated the coronary artery for MB and coronary atherosclerosis stenosis (CAS) in LAD and made a diagnosis by consensus. Significant independent risk factors for CAS were investigated by multivariate logistic regression analysis.

RESULTSA total of 3 182 (32.3%) cases of MB and 3 359 cases of CAS of LAD were identified. No patient with CAS in the tunneled segment was found. The mean length of bridges and the mean thickness of the overlying myocardium was (17.3±5.2) mm and (1.2±0.9) mm, respectively. There were 1658 MB cases in 3 359 cases of LAD stenosis and 1 524 MB cases in 6 503 cases of no LAD stenosis (χ(2)=681.12, P<0.05). Logistic regression analysis showed that MB in the LAD were significantly associated with CAS in the proximal LAD (OR=3.07, 95%CI=2.81-3.37, P<0.001), and after final adjustment for age, gender, body mass index, family history of heart attack, smoking, hypertension, dyslipidemia, diabetes mellitus, and resting heart rate (OR=2.86, 95% CI=2.60-3.16, P < 0.001).

CONCLUSIONMB in the LAD is independently associated with CAS in the proximal segment to MB.

Adult ; Aged ; Aged, 80 and over ; Coronary Stenosis ; complications ; diagnostic imaging ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Myocardial Bridging ; complications ; diagnostic imaging ; Risk Factors ; Tomography, X-Ray Computed