The culture medium and cultural conditions of solid-state fermentation of Streptomyces Menmyco-93-63 were tested in this study. The suitable medium which contains rice, sorghum, millet bran, and rice hull with the proportion of 2:2:3:3 was developed for the spore production of Streptomyces Men-myco-93-63 using single substrate screening, mixture substrate screening and orthogonal experiments, and the sporulation was up to 2.52?109 CFU/g. And then, initial charge, initial ratio of water to solid, inoculating quantity, and culture temperature impact to sporulation of Streptomyces Men-myco-93-63 were tested. The favorite cultural conditions are developed as the following: the initial charge is 15 g in 500 mL Erlenmeyer flask; initial ratio of water to solid is 1.7:1.0 (V/W, rice hull excluding), inoculating quantity is 7 mL, culture temperature is 28℃.

Objective To investigate clinical features and antibiotic resistance of pseudomonas aeruginosa (PA) strains isolated from patients with diabetic foot infections (DFI) in Tianjin Metabolic Diseases Hospital.Methods Eighty-five PA strains were isolated from 428 patients with diabetic foot in the hospital from Jan 2008 to Dec 2010.The clinical features of patients were summarized.Relationships between the isolates and depth of ulcer or severity of infection were analyzed.The disk-diffusion method was performed to examine antimicrobial susceptibility.Results Gram positive (G+) and Gram negative (Gˉ) isolates were 50.47％ and 41.12％,respectively.Multidrug-resistant PA composed 32.9％ of the total PA isolates.The size of ulcers with PA infections was bigger than those with non-PA bacterial infections (P＜0.05).Compared to G+ strains,patients with PA strains were older,had lower hemoglobin,but higher serum sensitive C-reactive protein; and more frequently,they had ischemic ulcer and osteomyelitis.Compared to G+ strains,the PA strains were more frequently isolated from deeper ulcers and with more serious infections(P＜0.05).The resistant rates of PA to cephalosporins,fluoroquinolones,and aminoglycosides were between 32.9％-61.2％,37.6％-42.4％,and 37.6％-62.4％,respectively.Only one out of 85 PA strains was imipenem-resistant.However,sensitiveness of all PA isolates to cefoperazone and sulbactam reached 100％.Conclusion PA strains are mainly found in patients with deeper ulcers and more serious infections.Multidrug-resistant PA is common in DFI.It is important to isolate pathogens and determine their antibiotic resistance correctly in diabetic foot patients in order to provide appropriate drug administration and to reduce the production and dissemination of drug resistant strains.

As an important part of Chinese medicinal materials, uncommon-territorial herbs are also the most complex parts in the herbal medicine markets. Through years of investigation on the key markets of Chinese herbal medicine, the meaning of uncommon-territorial herbs, their historical evolution, origin and characteristics were clarified in this paper, and some countermeasures were put forward for its development.
Biological Evolution ; China ; Drugs, Chinese Herbal ; chemistry ; history ; Herbal Medicine ; history ; History, 20th Century ; History, 21st Century ; History, Ancient ; Medicine, Chinese Traditional ; history ; Plants, Medicinal ; chemistry ; growth & development

OBJECTIVETo explore the method of constructing tissue-engineered skin using melanocytes and bone marrow mesenchymal stem cells (BMSCs) in vivo.

METHODSMelanocytes were isolated from human foreskin. BMSCs were isolated from human bone marrow. Both of them were co-cultured at a ratio of 1:10, and then were implanted into the collagen membrane to construct the tissue-engineered skin, which was applied for wound repair in nude mice. The effectiveness of wound repair and the distribution of melanocytes were evaluated by morphological observation, in vivo 4,6-diamidino-2-phenylindole, dihydrochloride (DAPI) fluorescent staining tracing, HE staining, S-100 immunohistochemistry, and transmission electron microscopy.

RESULTSThe wounds were satisfactorily repaired among the nude mice. The melanocytes were distributed in the skin with normal structure, as confirmed by DAPI fluorescent staining tracing, HE staining, S-100 immunohistochemistry, and transmission electron microscopy.

CONCLUSIONMelanocytes and BMSCs, after proper in vitro culture at an appropriate ratio, can construct the tissue-engineered skin with I type collagen membrane.

Animals ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Coculture Techniques ; Collagen Type I ; Humans ; Melanocytes ; cytology ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Nude ; Skin ; injuries ; Skin, Artificial ; Tissue Engineering

Professor LUO Cai-gui's experience of acupuncture at acupoint "Baliao" with twisting manipulation for treatment of low back pain is introduced. This method has significant efficacy on improving low back pain and numbness of lower extremities, which is characterized with short-time manipulation, quick de-qi and long effective time. The acupuncture methods, manipulations, precautions, etc. are elaborated in details. A typical case is added.
Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Humans ; Low Back Pain ; therapy ; Male ; Middle Aged

Aged ; Coronary Disease ; complications ; Humans ; Hypertension ; complications ; Renal Artery Obstruction ; diagnosis ; Renal Insufficiency ; complications ; Risk Factors

CD62p expression was an important monitoring parameter for preserved platelets quality. To setup an optimized flow cytometry assay for preserved platelets based on the CD62p expression on platelets, the platelet samples were collected, 0.1 mmol/L persantine and 1.1 mmol/L EDTA were added into the modified TB S used to replace PBS dilution; the methodological evaluation were carried out. Results showed that 0.1 mmol/L persantine and 1.1 mmol/L EDTA achieved to prevent platelets activation during the test procedure. The favorable negative or positive samples were prepared for check of fluorescence antibody's quality to ensure the validity of results. CD61 was used to identify platelets for assay and improve veracity of assay. The special injector was also replaced by special big syringe needle for blood collection to reduce in vitro artifacts, and the prepared sample can be steady-going for 48 hours at 4 degrees C after fixed by 1% paraformaldehyde. It is concluded that this flow cytometry assay for CD62p positive platelets is simple and efficient.
Blood Platelets ; chemistry ; Blood Preservation ; Flow Cytometry ; methods ; Humans ; P-Selectin ; blood

BACKGROUNDTubulointerstitial renal fibrosis is the common end point of progressive kidney diseases, and tubular epithelial-myofibroblast transdifferentiation (TEMT) plays a key role in the progress of tubulointerstitial renal fibrosis. Anaphylatoxin C3a and C5a are identified as novel profibrotic factors in renal disease and as potential new therapeutic targets. The aim of this study was to investigate whether C3a, C5a can regulate TEMT by transforming growth factor-β1 (TGF-β)/connective tissue growth factor (CTGF) signaling pathway and the effects of C3a and C5a receptor antagonists (C3aRA and C5aRA) on C3a- and C5a-induced TEMT.

METHODSHK-2 cells were divided into C3a and C5a groups which were subdivided into four subgroups: control group, 10 ng/ml TGF-β1 group, 50 nmol/L C3a group, 50 nmol/L C3a plus 1 µmol/L C3aRA group; control group, 10 ng/ml TGF-β1 group, 50 nmol/L C5a group, 50 nmol/L C5a plus 2.5 µmol/L C5aRA group. TGF-β1 receptor antagonist (TGF-β1RA) 10 µg/ml was used to investigate the mechanism of C3a- and C5a-induced TEMT. Electron microscopy was used to observe the morphological changes. Immunocytochemistry staining, real-time PCR and Western blotting were used to detect the expressions of a smooth muscle actin (α-SMA), E-cadherin, Col-I, C3a receptor (C3aR), C5aR, CTGF and TGF-β1.

RESULTSHK-2 cells cultured with C3a and C5a for 72 hours exhibited strong staining of α-SMA, lost the positive staining of E-cadherin, and showed a slightly spindle-like shape and loss of microvilli on the cell surface. The expressions of α-SMA, E-cadherin, Col-I, C3aR, C5aR, TGF-β1 and CTGF in C3a- and C5a-treated groups were higher than normal control group (P < 0.05). C3aRA and C5aRA inhibited the expressions of α-SMA, Col-I, C3aR, C5aR, and up-regulated the expression of E-cadherin (P < 0.05). TGF-β1 and CTGF mRNA expressions induced by C3a and C5a were partly blocked by TGF-β1RA (P < 0.05).

CONCLUSIONC3a and C5a can induce TEMT via the up-regulations of C3aR and C5aR mRNA and the activation of TGF-β1/CTGF signaling pathway in vitro.

Blotting, Western ; Cadherins ; genetics ; Cell Line ; Cell Transdifferentiation ; drug effects ; Complement C3a ; pharmacology ; Complement C5a ; pharmacology ; Epithelial Cells ; cytology ; drug effects ; ultrastructure ; Humans ; Immunohistochemistry ; Microscopy, Electron, Scanning ; Myofibroblasts ; cytology ; drug effects ; ultrastructure ; Real-Time Polymerase Chain Reaction

BACKGROUNDInnovative advancements in ultrasound instrumentation present a number of imaging modalities for myocardial contrast echocardiography (MCE) in ischemic syndromes. How well they compare to each other in diagnostic accuracy in the detection of acute myocardial infarction is unclear. The purpose of this study was to assess the relative accuracy of 3 different imaging modes of MCE, low mechanical index (MI) real-time perfusion imaging (RTPI), triggered harmonic angio mode (HA), and ultraharmonic imaging mode (UH) in the detection of acute experimental myocardial infarction within the time frame suitable for potential reperfusion.

METHODSMCE was performed in 10 open-chest dogs using RTPI, triggered HA and triggered UH modes at baseline and one hour after occlusion of left anterior descending coronary artery. Presence or absence of perfusion defects, and the perfusion defect size when present, were analyzed and compared with the infarct size delineated by triphenyltetrazolium chloride (TTC) staining.

RESULTSThe infarct area was (15.8 +/- 2.4)% by TTC staining; Perfusion defect area by MCE was similar to anatomic infarct area in all the three MCE approaches: (16.1 +/- 2.7)% by RTPI mode, (15.5 +/- 2.9)% by HA mode, and (15.5 +/- 3.0)% by UH mode. The sensitivity, specificity and overall diagnostic accuracy in the detection of myocardial infarction were 100%, 88%, and 94% for RTPI mode, 88%, 100%, and 94% for HA mode, and 100%, 75%, and 88% for UH mode.

CONCLUSIONAll modes of MCE, RTPI, triggered HA mode and triggered UH mode have excellent diagnostic accuracy in the immediate hour of acute coronary occlusion within the optimal time frame suitable for reperfusion therapy.

Animals ; Contrast Media ; Dogs ; Echocardiography ; methods ; Myocardial Infarction ; diagnostic imaging ; Staining and Labeling ; Tetrazolium Salts

OBJECTIVETo explore the potential application of IFN-gamma enzyme-linked immunospot (ELISPOT) assay in the diagnosis of tuberculous epididymitis (TE) by comparing ELISPOT assay with the traditional purified protein derivative (PPD) tuberculin skin test.

METHODSWe examined 13 TE patients using an in-house ELISPOT kit, another 11 TE patients by PPD skin testing, and 57 healthy male volunteers by parallel test with both the methods.

RESULTSTwelve (92.3%) of the 13 TE cases were positive on ELISPOT assay, and 10 (90.9%) of the 11 TE cases positive on PPD skin test, with no statistically significant differences between the two groups (P > 0.05). Among the 57 healthy male volunteers, 8 (14.0%) were positive on ELISPOT, and 28 (49.1%) positive on PPD test, the latter significantly higher than the former (P < 0.001).

CONCLUSIONIn terms of sensitivity, ELISPOT assay is similar to PPD test in the examination of tuberculous epididymitis. As for specificity, ELISPOT assay seems better than PPD test in differentiating tuberculous epididymitis patients from healthy males.

Adult ; Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Epididymitis ; diagnosis ; Humans ; Interferon-gamma ; Male ; Tuberculin ; Tuberculin Test ; Tuberculosis, Male Genital ; diagnosis