OBJECTIVELocalization of tracheal stem cells in rat trachea.

METHODSExtracorporeal tracheal injury (Wistar rats) was induced by 5-FU. The process of regeneration was observed and analyzed by light microscopy, electron microscopy, and immunohistochemistry.

RESULTSTwelve hours after treatment with 5-FU, the tracheal epithelium shed and cells with naked nuclei were seen located sparsely on the basement membrane. Six hours after removal of 5-FU, the tracheal rings were covered with flattened epithelium. These cells were poorly differentiated under electron microscopy. Immunohistochemistry showed few proliferating cell nuclear antigen (PCNA)-negative cells sparsely scattered among PCNA-positive cells on the basement membrane. Nine hours later, electron microscopy found that these cells differentiated into mucous cells and ciliated cells. Forty-eight hours later, the tracheal rings were entirely covered by pseudostratified ciliated columnar epithelium.

CONCLUSIONSA small number of G(0) cells with naked nuclei are located sparsely on the basement membrane of the trachea. Tracheal epithelium regenerates by proliferation and differentiation of these cells. It is likely that some of these G(0) cells on the tracheal basement membrane represent tracheal stem cells.

Animals ; Cell Differentiation ; Cell Proliferation ; Epithelium ; injuries ; ultrastructure ; Female ; Fluorouracil ; Male ; Microscopy, Electron ; Proliferating Cell Nuclear Antigen ; metabolism ; Rats ; Rats, Wistar ; Regeneration ; Stem Cells ; Trachea ; injuries ; pathology ; physiology ; Wound Healing ; physiology

The deubiquitinating enzyme ubiquitin specific peptidase 15 (USP15) is regarded as a regulator of TGFβ signaling pathway. This process depends on Smad7, the inhibitory factor of the TGFβ signal, and type I TGFβ receptor (TβR-I), one of the receptors of TGFβ. The expression level of USP15 seems to play vital roles in the pathogenesis of many neoplasms, but so far there has been no report about USP15 in psoriasis. In this study, immunohistochemical staining of USP15, TβR-I and Smad7 was performed in 30 paraffin-embedded psoriasis specimens and 10 normal specimens to investigate the expression of USP15, TβR-I and Smad7 in psoriasis and to explore the relevance among them. And USP15 small interfering RNA (USP15 siRNA) was used to transfect Hacat cells to detect the mRNA expression of TβR-I and Smad7. Of 30 cases of psoriasis in active stage, 28, 24 and 26 cases were positive for USP15, TβR-I and Smad7 staining, respectively. The positive rates of USP15 and Smad7 were significantly higher in psoriasis specimens than in normal skin specimens (44.1%±26.0% vs. 6.1%±6.6%, 47.2%±27.1% vs. 6.6%±7.1%), and positive rate of TβR-I (20.3%±22.2%) in psoriasis was lower than that in normal skin specimens (46.7%±18.2%). There was a significant positive correlation between USP15 and Smad7 expression, and significant negative correlations between USP15 and TβR-expression, an I d between TβR- and Smad7 expression I in psoriasis. After transfection of USP15 siRNA in Hacat cells, the expression of TβR-mRNA was up I -regulated and that of Smad7 was down-regulated. It is concluded that USP15 may play a role in the pathogenesis of psoriasis through regulating the TβR-I/Smad7 pathway and there may be other cell signaling pathways interacting with USP15 to take part in the development of psoriasis.

OBJECTIVETo investigate the relationship of serum metalloproteinase with the severity of liver fibrosis and inflammation.

METHODSA total of 88 patients with HBV-related liver fibrosis and early cirrhosis were enrolled from six hospitals. Serum fibrosis markers including hyaluronic acid (HA), IV collagen (IV-C), aminoterminal propeptide of type III procollagen (PIIIP), laminin (LN), matrix metalloproteinases (MMP) 1, 2, 9 and tissue inhibitors of metalloproteinase (TIMP) 1, 2 levels were determined. Liver biopsies were assessed according to a modified Scheuer and Chevallier's scoring system.

RESULTSSerum TIMP1 (r=0.540) and MMP2 (r=0.314) were correlated positively with the degree of hepatic fibrosis, whereas serum MMP1 (r=-0.495) was correlated negatively. By receiver operating curve analysis (ROC), the sensitivity to distinguish the fibrosis stage 2 from stage 1 was 90.5% and the specificity was 52.0% if the cut-off value of MMP1 was 13.96 ng/ml, and the sensitivity was 91.6% and the specificity was 64.0% if the cut-off value of TIMP1 was 76.84 ng/ml. The sensitivity to distinguish cirrhosis (stage 4) from fibrosis (stage 3) was 70.7% and specificity was 80.9% if the cut-off value of MMP1 was 6.86 ng/ml, and the sensitivity was 60.5% and the specificity was 92.3% if the cut-off value of TIMP1 was 210.04 ng/ml.

CONCLUSIONSerum TIMP1, MMP1, MMP2 levels and TIMP1/MMP1 ratio could be used as serum fibrosis markers.

Adult ; Biomarkers ; blood ; Female ; Hepatitis B, Chronic ; blood ; complications ; Humans ; Liver Cirrhosis ; blood ; virology ; Male ; Matrix Metalloproteinase 1 ; blood ; Matrix Metalloproteinase 2 ; blood ; Middle Aged ; Tissue Inhibitor of Metalloproteinase-1 ; blood

The 11-hydroxylation of 16alpha,17alpha-epoxy-4-pregenene-3,20-dione as a useful intermediate for the preparation of hormones can be achieved by the mycelium of Absidia coerulea at higher conversion rate than using other strains. In this paper 16alpha,17alpha-epoxy-4-pregenene-3,20-dione mixed with a little water, beta-cyclodextrin, Tween-80 was introduced into the fermentation broth after ultrasonication to increase pseudo-water-solubility of the hydrophobic substrate. This pseudo-crystallo feed could avoid the toxicity of organic solvents and was more available for the microbial transformation. The multi layer feed-forward neural network was used to setup a model which indicated the relationship between medium and feed components and the conversion rate. Particle swarm optimization (PSO), which was a stochastic global optimization algorithm and of which the convergence speed was high, was applied to obtain the optimal concentration of the medium and feed components. At optimum conditions with the pseudo-crystallo feed, the conversion rate of 16alpha,17alpha-epoxy-4-pregenene-3,20-dione at an initial concentration of 10 g/L was 87.5% in shaking flasks. The conversion rate of the substrate was up to 86.6% at higher concentration of 20 g/L feed in a 3.7 L fermentor.
Absidia ; metabolism ; Fermentation ; Hydroxylation ; Pregnenediones ; metabolism

With the emergence of drug resistant tuberculosis, it is very urgent to find novel anti-tuberculosis drugs, especially novel anti-drug-resistant tuberculosis drugs. Because of the slow growth and the need to work in a biosafty environment of Mycobacterium tuberculosis, the development of evaluation of drug effect is severely impeded. In order to solve these issues, non-pathogenic fast-growing Mycobacterium smegmatis is introduced as test organism. The inhA is one of a target of isoniazid (INH) overexpression or mutation of this gene in Mycobacterium tuberculosis conferring resistant to INH. A recombinant plasmid bearing inhA was constructed and electroporated into Mycobacterium smegmatis, using shuttle expression vector pMV261. Transformants were induced to express a protein of inhA, identified by SDS-PAGE. Results show that Mycobacterium smegmatis containing inhA plasmids exhibited 100-fold or greater increased resistance to INH, but it conferred no increased resistance to others first-line anti-tuberculosis drugs. Resazurin microtiter assay plate testing of Mycobacterium smegmatis susceptibility to drugs is a rapid, simple, and inexpensive method and could decrease color background of drugs by detecting fluorescence. It will be benefit for high-throughout screening of drugs of anti-isoniazid-resistant Mycobacteria.

The HPLC method was established to simultaneously determine the contents of myricetin, luteolin, apigenin and kaempferol in Wikstroemia indica ( L. ) C. A. Mey. The method was carried out on a Diamonsil C18 column (4. 6 mm x 250 mm, 5 µm) eluted with the mobile phases of water containing 0.15% phosphoric acid and acetonitrile in gradient mode. The UV detection wavelength was 365 nm. The flow rate was 1.0 mL · min(-1) and the column temperature was set at 30 °C. All the standard compounds showed a good linearity in the range of 0.100 8-1.008 (r = 0.999 2), 0.484 8-4.848 (r = 0.999 0) , 1. 354-13. 54 (r = 0.999 6), 0.316 8-3.168 mg · L(-1) (r = 0.999 0) for myricetin, luteolin, apigenin and kaempferol, respectively. The average recoveries of these four flavonoids were 98.5%, 100.9%, 99.7% and 98.9% with RSD 1.2%, 1.7%, 0.81% and 1.6%, respectively. In conclusion, the method is simple, rapid and accurate. It can be applied for the quality control of Wikstroemia indica.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Flavonoids ; analysis ; Wikstroemia ; chemistry

Hemangioma ; pathology ; Humans ; Joint Diseases ; pathology ; Male ; Middle Aged ; Synovial Membrane ; pathology

Objective To investigate the effects of hyperbaric oxygen (HBO) and right median nerve electrical stimulation (RMNS) on the coma arousal therapy in patients with severe traumatic brain injury. Methods Eighty-three patients with severe traumatic brain injury on the recovery phase were randomly divided into control group (n=41) and treatment group (n=42). The patients in the control group were given such routine therapies as reducing the intracranial pressure and giving nutrition to the nerves and conventional rehabilitation nursing. The patients in the treatment group also treated with HBO and RMNS. Glasgow coma scale (GCS), Glasgow outcome scale (GOS), activities of daily living (ADL)scale, European stroke scale (ESS) and Kamofsky performance scale (KPS) were performed on these patients before, 1 and 3 after the treatment. The cerebral blood flow (CBF) and the electrical activity were detected by TCD and brain electrical activity mapping (BEAM). Results As compared with thecontrol group, the treatment group got significant improvement on each scale in the 1st and 3rd month oftreatment (P＜0.05). CBF in the treatment group was obviously increased, while that in only 17 patients in the control group was increased. BEAM showed that the diffused inactive echoes were significantly decreased in the treatment group as compared with those in the control group. Conclusion HBO andRMNS can promote brain tissue rehabilitation and accelerate recovery of brain function in patients with severe traumatic brain injury, thereby contributing to the improvement of the level of awareness.

OBJECTIVETo study the viremia formation in guinea-pigs infected with wild type and attenuated Japanese encephalitis virus (JEV).

METHODSGuniea pigs were inoculated intraperitoneally with different wild JEV strains and the attenuated vaccine strain and its parent virulent strain. Viremia was detected on different days following virus inoculation.

RESULTSAll the guinea-pigs inoculated with the wild JEV strains induced different levels of viremia (1.00-3.40 Lg pfu) on the 1st and 3rd day post inoculation. Using a virus titer of 10(4) pfu for inoculation, the animals inoculated with the SA14 parent strain induced relatively high viremia (10(2.4)-10(3.4) pfu), however no viremia coulds be detected on any tested days.

CONCLUSIONThe degree of viremia in guinea pigs can be used as a new method to evaluate the attenuation of JEV.

Animals ; Disease Models, Animal ; Encephalitis Virus, Japanese ; pathogenicity ; physiology ; Encephalitis, Japanese ; virology ; Guinea Pigs ; Humans ; Japanese Encephalitis Vaccines ; administration & dosage ; adverse effects ; Vaccines, Attenuated ; administration & dosage ; adverse effects ; Viremia ; virology ; Virulence ; Virus Replication

Objective To investigate the effect of comprehensive intervention measures on submission of microbiological specimens before therapeutic use of antimicrobial agents.Methods August 2014 to August 2015 was as preintervention group,comprehensive intervention measures were started to carry out from September 2015,Septem ber 2015 to September 2016 was as post-intervention group.Microbiology specimen submission rates before therapeutic antimicrobial use,positive rate of blood culture,and rate of antimicrobial use in all departments and key departments were compared between pre-intervention group and post-intervention group.Results The submission rates of microbiological specimens before general,restricted,and special therapeutic antimicrobial use increased from 42.21％,45.19％,and 74.71％ of pre-intervention respectively to 53.54％,55.68％,and 89.70％ of postintervrntion respectively,showing significant difference (all P＜0.01);after intervention,except gastrointestinal surgery and trauma department of orthopedics,the other departments all met the requirements of the microbiological specimen submission rates set by the hospital;but submission rates of microbiological specimens from department of gastrointestinal surgery and trauma department of orthopedics increased from 5.46％ and 11.67％ before intervention to 11.66％ and 29.45％ respectively after intervention,difference was statistically significant (both P＜0.001).The missing report rate of healthcare-associated infection(HAI) dropped from 13.56％ before intervention to 10.98％ (P＜ 0.05),and the use rate of antimicrobial agents decreased from 57.36％ to 54.47％ (P＜0.001).Conclusion Comprehensive intervention measures can effectively improve the submission rates of microbiological specimens before therapeutic use of antimicrobial agents,reduce missing report rates of HAI and utilization rate of antimicrobial agents,and achieve certain clinical effectiveness.