Aged, 80 and over ; Dyspnea ; etiology ; Glottis ; pathology ; Hematoma ; complications ; Hemophilia A ; complications ; Humans ; Laryngeal Diseases ; complications ; Male

Objective: in order to provide rapid and reliable method. Methods: Encoded Annexin Ⅴ cDNA was amplifyed from U937 cDNA libary by PCR and then subcloned into E coli expression vector. MS2-Annexin Ⅴ fusion protein could be overexpressed in E　coli. The MS2 bacteria protein could be removed by thrombin digestion.The mature Annexin Ⅴ was obtained by ion exchange chromatography and the FITC labled Annexin Ⅴ could be used in the detection of apoptosis. Results:Up to 37% of the total bacterial proteins was rhAnnexin Ⅴ as showed by SDS-PAGE. The purification of Annexin Ⅴ is over 99%. The FITC labled Annexin Ⅴ could efficiently detect apoptosis. Conclusion: We successfully established the technique procedure of obtaining a large quantity of Annexin Ⅴ and provided the basic routine for popularizing the detection of apoptosis' with high effciency.

To determine the distribution and drug resistance tendency of Gram-negative bacillus so as to provide a reasonable selection for antibiotic in clinical treatment.The clinical isolates bacillus were mainly Gram-Negative bacillus during three years.The detection out ratio for Gram-Negative bacillus had decreased gradually but the ratio for non-fermental gram-negative bacillus had increased.The combinations of antibiotic and enzyme inhibitor has more antibacterial activity to non-fermental gram-negative bacillus than antibiotic.The detection out ratio for ESBLs had gradually increased.

OBJECTIVETo compare the contents of resveratrol and polydatin in some materials of Polygonum cuspidatum from various sources, so to screen and obtain the suitable cultures for the following metabolism regulation study.

METHODRP-HPLC method was applied to simultaneously assay resveratrol and polydatin in different samples.

RESULTBy the modified methods of extraction and determination, large amount of materials were screened. The results indicated that the contents of resveratrol and polydatin in root and rhizome were evidently higher than those in the leave and stems. The content of polydatin in the seedlings cultured indoor for three months was 1.27% and showed a 1.25-time increse than that in the wild plants, while the content of resveratrol (0.401%) approached that in the wild plants. Both of resveratrol and polydatin could be examined from different tissue cultures of P. cuspidatum, such as the sterile seedlings, callus, suspended cells and hairy roots, and the levels of them were closely related to the growth speed, physiological status and developmental phase. Hairy roots had the highest potentiality in several tested cultures and the increase rate of dry weight was 8.29 when cultured in vitro for 30 days, and showed a 8.4-fold and a 192.8-fold increase compared with those of natural roots and suspended cells, respectively. The content of polydatin in the hairy roots was up to 0.037% and that of resveratrol was 0.007%.

CONCLUSIONThe established analysis method is rapid, simple and accurate, especially adapted to the simultaneous determination of resveratrol and polydatin in massive biological samples. Hairy-root cultures have the superiority among the tested materials of P. cuspidatum and are suitable for the large-scale biomass and consistent production of efficient constituents.

Biomass ; Chromatography, High Pressure Liquid ; methods ; Fallopia japonica ; chemistry ; growth & development ; Glucosides ; analysis ; Plant Leaves ; chemistry ; growth & development ; Plant Roots ; chemistry ; growth & development ; Plant Stems ; chemistry ; growth & development ; Plants, Medicinal ; chemistry ; growth & development ; Reproducibility of Results ; Rhizome ; chemistry ; growth & development ; Seedlings ; chemistry ; growth & development ; Stilbenes ; analysis ; Tissue Culture Techniques ; methods

Adult ; Cartilage Diseases ; diagnosis ; etiology ; Female ; Humans ; Knee Joint ; surgery ; Menisci, Tibial ; surgery

BACKGROUNDAtopic dermatitis (AD) is characterized by defective skin barrier and imbalance in T helper 1/T helper 2 (Th1/Th2) cytokine expression. Filaggrin (FLG) is the key protein to maintaining skin barrier function. Recent studies indicated that Th1/Th2 cytokines influence FLG expression in keratinocytes. However, the role of Th1/Th2 cytokines on FLG processing is not substantially documented. Our aim was to investigate the impact of Th1/Th2 cytokines on FLG processing.

METHODSHaCaT cells and normal human keratinocytes were cultured in low and high calcium media and stimulated by either interleukin (IL)-4, 13 or interferon-γ (IFN-γ). FLG, its major processing proteases and key protease inhibitor lymphoepithelial Kazal-type-related inhibitor (LEKTI) were measured by both real-time quantitative polymerase chain reaction and Western blotting. Their expression was also evaluated in acute and chronic AD lesions by immunohistochemistry.

RESULTSIL-4/13 significantly reduced, while IFN-γ significantly up-regulated FLG expression. IL-4/13 significantly increased, whereas IFN-γ significantly decreased the expression of kallikreins 5 and 7, matriptase and channel-activating serine protease 1. On the contrary, IL-4/13 significantly decreased, while IFN-γ increased the expression of LEKTI and caspase-14. Similar trends were observed in AD lesions.

CONCLUSIONSOur results suggested that Th1/Th2 cytokines differentially regulated the expression of major FLG processing enzymes. The imbalance between Th1 and Th2 polarized immune response seems to extend to FLG homeostasis, through the network of FLG processing enzymes.

Caspase 14 ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Dermatitis, Atopic ; metabolism ; Humans ; Immunohistochemistry ; Interferon-gamma ; metabolism ; Interleukin-13 ; metabolism ; Interleukin-4 ; metabolism ; Intermediate Filament Proteins ; metabolism ; Keratinocytes ; enzymology ; metabolism ; Proteinase Inhibitory Proteins, Secretory ; metabolism ; Serine Peptidase Inhibitor Kazal-Type 5 ; Th1 Cells ; metabolism ; Th2 Cells ; metabolism

OBJECTIVESTo study the effects of hepatectomized rat serum and hepatocyte growth factor (HGF) on the transdifferentiation of adult rat bone marrow stem cells (ABMSCs) into hepatic parenchymal cells.

METHODSThe serum was collected from the rats 24 hours after being subjected to subtotal hepatectomy. ABMSCs were collected and cultured in DMEM/F12 (1:1) containing the hepaetectomized rat serum or HGF. The differentiated hepatocyte-like cells were labeled with CM-DiI and administrated by tail vein injection into the isogeneic rats. The cultured and injected cells were both identified by immunocytochemistry and cultured cells were assayed using RT-PCR and Western blot.

RESULTSHepatectomized rat serum and HGF were demonstrated to have the effect of inducing transdifferentiation of ABMSCs into hepatocyte-like cells in vitro. The differentiated cells expressed albumin mRNA and albumin after 7 days++'s co-incubation. Albumin-expressing and CM-DiI positive hepatocyte-like cells were characterized in livers and spleens of the rats injected with the cultivated cells.

CONCLUSIONABMSCs could transdifferentiate into hepatic parenchymal cells by hepatectomized rat serum or HGF.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Culture Media ; Hepatectomy ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; Liver Regeneration ; Rats ; Rats, Sprague-Dawley ; Serum ; Stem Cells ; cytology

OBJECTIVETo study the rabies molecular biology features in animals between high incidence area of rabies and no rabies cases area in Hunan.

METHODSdetect saliva of dogs and brains of dogs and cats by direct immunofluorescence assay, review positive samples by RT-PCR, sequencing extract RNA virus for genetic analysis.

RESULTS12 were detected rabies virus antigen and positive nucleoside acid in 82 dogs from Wugang city also 1 in 17 from Dongkou county; the positive rate: Wugang 14.63 percent, Dongkou 5.88 percent. No rabies virus was detected in 67 samples of dog brains from Fenghuang County. Also none in 28 samples of cat brains. Amplificating N gene of rabies virus from positive samples of dog brain's tissue (No Wg13, Dk13) by RT-PCR, it shows that homology of nucleoside acid between two strain of virus is 99.4 percent; also 99.1 percent of amino acid. The homology of nucleoside acid (amonio acid) among Wg13 stain and Chinese strain CTN and aG strain is 89.4 percent (98.2 percent) and 86.1 percent (95.1 percent); The homology of nucleoside acid (amonio acid) among Dk13 stain Chinese strain CTN and aG strain is 89.1 percent (98.0 percent), 86.1 percent (94.9 percent).Compare with isolated rabies virus from abroad, the homology between two strains and Indonesia is 92.8 percent and 93.2 percent, the most similar of them. The strains isolated from other countries including Japan, Sri Lanka and India are relatively lower; The sequence of gene Wg13 and Dk13 were taken replacement of amino acid.

CONCLUSIONTwo strains are belong to type I rabies virus, comparing its N gene with current using vaccine strains, both are in same group, and homology are relatively higher.

Animals ; Cats ; Dogs ; Genes, Viral ; Phylogeny ; Rabies virus ; classification ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective:To investigate the surgical treatment of the tumors at cervicothoracic junction.Methods:A retrospective analyses was performed for 63 patients with tumors at the cervicothoracic junction receiving surgery from Mar 2008 to May 2020 in the Department of Thoracic Surgery, Zhongshan Hospital, Fudan University. Clinical data about manifestation, surgical approach, resection degree and pathological types were collected. There were 43 cases of asymptomatic patients and 20 cases of patients with ≥1 clinical manifestations. Twenty two patients receiving radical resection with video-assisted thoracoscopic surgery. Anterior approach was the most popular treatment in open surgery (24 cases, 38.1%), and 8 cases of anterolateral approach(6 cases of Hemiclamshell incisions, 2 cases of trap-door incisions), 1 case of posterior approach, 2 cases of posterolateral approach and 1 case of supraclavicular combined posterolateral approach.Results:Pathological examination suggested 61 cases of radical resection and 2 cases of microscopic residual. Neurilemmoma was the most common pathological type (27 cases, 42.9%), the second common pathological type was tumor originated from fibrous tissues (6 cases, 9.5%). The 3-year overall survival rate of those 63 patients was 88.9%, while the 5-year overall survival rate was 84.1%.Conclusion:Tumors involving the cervicothoracic junction are characterized as special location, complicated anatomy and various histopathological subtypes. Individualized approach and surgery improve safety and normalization of tumors at cervicothoracic junction treatment.

OBJECTIVETo explore the abnormal expressions of testicular reproduction-related genes induced by glycosides of Tripterygium wilfordii (GTW) and the intervention with kidney-tonifying Chinese herbs.

METHODSAdult Balb/C male mice were fed on GTW at 30 mg per kg per d for 3 weeks to establish a model of reproductive dysfunction. The model mice were divided into different groups to receive intragastrical administration of saline (0.25 ml/d), GTW (30 mg per kg per d), Cistanche (10 g per kg per d), Rehmannia (10 g per kg per d), and Rehmannia + Cistanche (20 g per kg per d), respectively, once a day for 3 weeks. And a Cistanches pretreatment group was treated with GTW (30 mg per kg per d) and Cistanche (10 g per kg per d) for the same length of time. Then we detected the changed expressions of testicular reproduction-related genes Dzip1, Fas, c-jun and Wnt4 in each group.

RESULTSThe model mice showed an obviously down-regulated expression of the Y chromosome microdeletion-related gene Dzip1, and up-regulated expressions of the germ cell apoptosis-related gene Fas, proto-oncogene c-jun, and signal transduction-related gene Wnt4. Intervention with Chinese herbs achieved different degrees of improvement of the mice's reproductivity, and the most obvious efficacy was observed with the combined use of kidney-yang tonifying Cistanche and kidney-yin nourishing Rehmannia.

CONCLUSIONGTW exerts significant impact on reproduction-related genes. Both the kidney-yang tonifying drug Cistanche and kidney-yin nourishing drug Rehmannia can counteract some of the reproductive toxicity of GTW, while the combination of the two can further enhance the effect.

Animals ; Cistanche ; Drugs, Chinese Herbal ; pharmacology ; Glycosides ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Testis ; drug effects ; metabolism ; Tripterygium ; chemistry