Optimization of surgical treatment of biliary dilatation (BD) depends on reasonable clinical classification and standardized classification-based treatment strategy.Due to increasing limits and defects of classic Todani classification,a new classification named Dong-classification has been proposed,which was based on a large series analysis from a single referral center.Some important parameters including anatomical location and range of BD,pathogenic factors,and different surgical managements were main considerations in the new classification.After practical application and evaluation,Dong-classification has been improved step by step.It is believed that Dong-classification may contribute to improving surgical treatment decision and selecting reasonable operative plan.

Objective To study the function of Bak in mitochondrial signaling pathway and interaction between Bax and Bak during apoptosis.Methods Bax/Bak double knock out(Bax-/- and Bak-/-)MEF cells from mouse embryo fibroblasts(MEF) and Hela cells were divided into four groups according the cell different genotypes(wild type,Bax-/-,Bak-/- and double knock out) and treated with different chemical reagents after co-transfection with Bax,Bak,Mito-Red and empty pEGFP vector.Apoptosis,mitochondrial morphology and cytochrome C release were detected with confocal microscope,immunofluoresence and Western blotting techniques.Results There were correlations between the percentage of Hela cell apoptosis and mitochondrial fission(%) as well as cytochrome C(%)(P

To investigate the components in Plastrum Testudinis which have effects on the proliferation of rat bone marrow mesenchymal stem cells( bMMSCs), the active parts of plastrum testudinis which can promote proliferation of rat mesenchymal stem cells were extracted by petroleum aether. The activities of inducing the proliferation of bMMSCs were studied by MTT assay and flow cytometry. The chemical components of extraction were analyzed by GC-MS. The results showed that the petroleum aether extraction can obviously promote the proliferation of the stem cells. The main components are long-chain fatty acids, cholesterols and cholest-4-en-3-one, and palmitic acid, stearic acid and cholest-4-en-3-one have effects on proliferation of bMMSCs. In plastrum testudinis, fatty acids can promote the proliferation of bMMSCs but not increase overly. This provide the experiment basis, and offer important reference for Traditional Chinese Medicine that researches stem cells.
Animals ; Bone Marrow Cells ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Fatty Acids ; isolation & purification ; pharmacology ; Materia Medica ; chemistry ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; Rats ; Turtles

Objective To apply special staining techniques in pathological diagnosis of fungal infections in HIV/AIDS patients.Methods Pathological data of 20 HIV/AIDS patients complicated with fungi infections in Shanghai Public Health Clinical Center during February 2010 and November 2013 were retrospectively analyzed.Tissue specimens were stained with hematoxylin and eosin (HE),Periodic acidSchiff (PAS) and methenamine silver nitrate (MSN),and the sections were observed under optical microscope.Results Among 20 HIV/AIDS patients complicated with fungi infections,2 were infected with pulmonary cryptococcosis; 3 were penicillium marneffei infections in skin,lung and abdominal mesenteric lymph nodes; 5 were histoplasma capsulatum infections in epiglottis,neck lymph nodes,oral cavity,abdominal cavity and skin; 4 were aspergillus infections in maxillary sinus,lung and vocal cords,and 3 of them were combined with tuberculous lesions; 6 were candida albicans infections in liver,pharynx,esophagus and stomach.In tissues stained with HE the infiltration of inflammatory cells,granuloma formation and coagulative necrosis were observed,and the shape of fungi needed careful observation to avoid missed diagnosis and misdiagnosis.In tissues stained with PAS,fungal spores and pseudohypha were presented in bright amaranth,and cell nucleus was in purple-blue.In tissues stained with MSN,fungal spores and pseudohypha were identified clearly in brown-black.Conclusion HE plus PAS and MSN staining will help pathological diagnosis of fungi infection.

Objective To construct a recombinant eukaryotic expression plasmid of glycosylphosphatidyl inositol(GPI)-anchored bcr/abl and explore its expression at mRNA and protein level.Methods The gene fragment encoding bcr/abl was amplified by PCR using the plasmid containing the cDNA sequence of P210 as template and then inserted into a eukaryotic expression vector pBudCE4.1.The constructed recombinant plasmid pBudCE4.1-bcr/abl was identified by restriction analysis and DNA sequencing.Lymphocytes were isolated from human peripheral blood and their total RNA was extracted.The gene fragment encoding GPI was amplified by RT-PCR using the obtained RNA as template and was inserted into the constructed recombinant plasmid pBudCE4.1-bcr/abl in order to anchor GPI and bcr/abl.The constructed recombinant plasmid pBudCE4.1-bcr/abl-GPI was transfected into COS-7 cells,and the expressions of objective fragment were detected by RT-PCR and Western blotting.Results The results of restriction analysis,PCR and DNA sequencing proved that GPI-anchored bcr/abl fusion fragment was correctly inserted into vector pBudCE4.1.The expression of bcr/abl fusion gene and fusion protein were identified in transfected COS-7 cells and on their membrane.Conclusion The recombinant plasmid pBudCE4.1-bcr/abl-GPI was successfully constructed and expressed on the membrane of COS-7 cells,which found a basis of cell immunity with GPI-anchored bcr/abl fusion gene.

BACKGROUNDIntratracheal instillation of blood induces self-repaired acute lung injury. However, the mechanism of repair has been unclear. Heme-oxygenase (HO)-1, which catalyzes heme breakdown, acts as an inducible defense against oxidative stress and plays an important role in inflammation. The objective of this study was to test the role of HO-1 in lung injury caused by intratracheal instillation of red cells.

METHODSForty healthy, male Sprague-Dawley rats were randomly divided into five groups: normal group, saline group, erythrocyte group, erythrocyte+zinc-protoporphyrin (ZnPP, HO-1 inhibitor) group and saline+ZnPP group. At 2 days after intratracheal instillation of red cells, lung tissues and lavage samples were isolated for biochemical determinations and histological measurements.

RESULTSHistological analysis revealed that administration of ZnPP worsened the acute lung injury induced by instilled erythrocytes. HO-1 was over-expressed in the erythrocyte group and in the erythrocyte + ZnPP group. Compared with the erythrocyte + ZnPP group, the levels of total protein, lactate dehydrogenase and tumor necrosis factor-alpha in the lavage were lower (P < 0.01), while the level of interleukin-10 was higher in the erythrocyte group (P < 0.01).

CONCLUSIONHO-1 protects against erythrocyte-induced inflammatory injury in lung.

Animals ; Erythrocytes ; physiology ; Heme Oxygenase (Decyclizing) ; analysis ; physiology ; Interleukin-10 ; analysis ; Lung ; pathology ; Lung Injury ; prevention & control ; Male ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; analysis

BACKGROUND:Multimodal analgesia provides sufficient analgesia in renal recipients and appears to be associated with the recovery of renal function after transplantation. OBJECTIVE:To investigate the effect of multimodal analgesia with dezocine on postoperative immunity after renal transplantation, and discuss the appropriate analgesic drugs and methods for patients with renal transplantation. METHODS:Forty patients undergoing renal transplantation were randomly divided into two groups. They al received general anesthesia combined with epidural blockage. Control group received intramuscular injection of analgesic drugs when needed, while dezocine group received multimodal analgesia:preemptive anaIgesia with dezocine+patient-control ed epidural analgesia. The heart rate, mean arterial pressure, and saturation of blood oxygen were detected before anesthesia, 12, 24, 48 hours after transplantation. T lymphocyte subsets, interleukin-2, interleukin-6 and interleukin-10 levels in venous blood were measured before anesthesia, 12, 24, 48 hours after transplantation. RESULTS AND CONCLUSION:Compared with before anesthesia, the CD4+, CD8+cellsubset counts, CD4+/CD8+ratio, the levels of interleukin-2 and interleukin-6 were decreased significantly (P<0.05), and the levels of interleukin were significantly increased after transplantation in the control group (P<0.05). The postoperative CD4+cellsubset counts, the levels of interleukin-2 and interleukin-6 were significantly lower at 12 hours after transplantation than that before anesthesia (P<0.05), then recovered to normal levels at 24 hours in dezocine group. The postoperative CD8+cellsubset counts, CD8+and CD4+/CD8+ratio were not changed before and after transplantation in the dezocine group. The levels of interleukin-10 in the dezocine group were significantly increased at 48 hours after transplantation compared with before anesthesia (P<0.05), which was stil lower than that in control group (P<0.05). Multimodal analgesia with dezocine can effectively protect the immune system, promote short-term turnover of renal function, and prolong graft survival for patients with renal transplantation.

In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.
Animals ; Half-Life ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; Mice ; Molecular Probes ; pharmacokinetics ; RNA, Small Interfering ; chemistry ; Rabbits

OBJECTIVETo explore the effects of noise exposure level and cumulative noise exposure (CNE) on the relationship between rs2070424 and rs10432782 SNPs in SOD1 and the susceptibility to noise-induced hearing loss (NIHL).

METHODSA case-control study was performed for investigating the effects of environmental risk factors on the susceptibility to NIHL in 201 sensitive workers and 202 resistant workers.A questionnaire was utilized to investigate the occupational health and to identify the occupational risk factors. The noise exposure levels were detected according to the Chinese standard Measurement of noise in the workplace (GBZ/T 189.8-2007). The peripheral blood samples (5 ml blood for each sample) were from sensitive workers and resistant workers. Genomic DNA was extracted on the basis of the standard procedures of Takara kit. SNPs were detected using standard procedures of TaqMan probe allele identification method.

RESULTSIn group exposed to 85 - 92 dB noise (A), the risk of NIHL in the subjects with the AA genotype of rs2070424 was lower than that in the subjects with the GG genotype, OR = 0.37 (95%CI: 0.17∼ 0.80). In group exposed to > 82 dB CNE (A), the AA genotype of rs2070424 is a protective factor of NIHL, as compared with the GG genotype, OR = 0.25 (95%CI: 0.09 ∼ 0.70). In group exposed to 85 - 92 dB noise (A), the risk of NIHL in the subjects with the GG genotype of rs10432782 was compared with the risk of NIHL in the subjects with the TT genotype, OR = 3.17 (95%CI: 1.16 ∼ 6.89). The GT genotype was compared with TT genotype, OR = 2.39 (95%CI: 1.16 ∼ 4.97). In group exposed to 75 ∼ 82 dB CNE (A), the risk of NIHL in the subjects with the GG genotype was compared with the risk of NIHL in the subjects with the TT genotype, OR = 2.35 (95%CI: 0.96 ∼ 5.72), P = 0.06. The GG genotype may bea risk factor of NIHJ.

CONCLUSIONThe noise exposure level and CNE may influence the relationship between rs2070424, rs10432782 SNPs in SOD1 and noise-induced hearing loss.

Adult ; Case-Control Studies ; Female ; Genetic Predisposition to Disease ; Genotype ; Hearing Loss, Noise-Induced ; etiology ; genetics ; Humans ; Male ; Noise, Occupational ; adverse effects ; Polymorphism, Single Nucleotide ; Superoxide Dismutase ; genetics ; Superoxide Dismutase-1 ; Surveys and Questionnaires

Objective To study the effects of selenium deficiency,iodine deficiency and combined selenium and iodine deficiency on bone and cartilage growth in the parental and the first filial generation rats. Methods Forty-eight weanling healthy SD rats were randomly divided into selenium deficieney, iodine deficiency, combined selenium and iodine deficiency and control groups according to their body mass. These rats were fed with selenium deficiency, iodine deficiency, combined selenium and iodine deficiency, and normal fodder, respectively. The parental rats (about 3 months old) were mated in each group 8 weeks after the beginning of the experiment. Right tibias and left knee joints were collected when the parental generation rats were about 6 months and the first filial generation rats were about 3 months old. Tibial length, mid-shaft tibial diameter, and articular cartilage diameters of the right tibias were measured by vernier caliper. Left knee joints were embedded and cut into sections and the thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes in growth plate cartilage were observed under the light microscope. Results The selenium deficiency had significant effect on serum selenium level of the parental and the first filial generation rats(F value were 239.56,232.68, P< 0.01), and also on serum T4 level of the first filial generation rats(F value were 6.95, P < 0.05). The iodine deficiency had significant effect on serum T3 and T4 level in the two generations rats(F value were 14.11,14.05,30.29,34.53, P < 0.01 ). There were interactions between selenium deficiency and iodine deficiency on serum T4 level in the first filial generation rats (F= 5.99, P< 0.05). The serum selenium of selenium deficiency group[ (30.28 ± 6.34), (43.95 ± 9.75)μg/L],combined selenium and iodine deficiency group[ (30.33 ± 5.18), (35.40 ± 3.16)μg/L] were significantly lower than iodine deficiency group[(345.83 ± 29.55), (245.24 ± 9.95)μg/L] and the controls[ (358.64 ± 30.50), (236.50 ±9.75) μg/L] in the two generations. The serum T3 of combined selenium and iodine deficiency group [(0.55 ± 0.05 ),(0.88 ± 0.14)nmol/L] were significantly lower than the controls[(0.75 ± 0.08), (1.26 ± 0.26)nmol/L] in the two generations. The serum T4 of iodine deficiency [ (24.11 ± 2.29), (42.10 ± 8.92) nmol/L ] and combined selenium and iodine deficiency group[ (20.66 ± 1.93), (26.55 ± 5.98)nmol/L] were significantly lower than the controls[ (36.15 ±2.74), (52.79 ± 8.84)nmol/L] and selenium deficiency group[ (28.12 ± 3.33), (52.02 ± ll.99)nmol/L] in the two generations. The selenium deficiency and iodine deficiency had significant effect on tibial length, thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes in first filial generation rats(F values were 24.31,6.98,40.76,56.15,25.24,82.82, 10.07,5.57, P <0.05 or <0.01). There were interactions between selenium deficiency and iodine deficiency on tibial length, thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes (F values were 5.68,24.86,41.82,9.12, P <0.05 or <0.01 ). The tibial length of the selenium deficiency group[ (33.17 ± 0.34)mm] and combined selenium and iodine deficiency group[ (31.30 ± 0.87)mm] were significantly lower than the controls[ (34.12 ± 0.32)mm, P< 0.05]. Thickness of the growth plate cartilage [ (1.60 ± 0.18)mm ], layers of proliferative chondrocyte (8.54 ± 0.81), and hypertrophic chondrocyte (4.95 ± 0.37)of the combined selenium and iodine deficiency group were significantly decreased when compared to the selenium deficiency group[ (3.03 ± 0.10)mm, 14.68 ± 0.84,6.60 ± 0.31], iodine deficiency group[ (2.90 ± 0.09)mm, 13.75 ±0.33,6.61 ± 0.84 ] and the controls [ (3.19 ± 0.09) mm, 14.94 ± 0.36, 6.64 ± 0.26, P <0.05]. Thickness of the growth plate cartilage, layers of proliferative chondrocyte of the iodine deficiency group were lower than the controls(P<0.05). Conclusions Selenium deficiency impair tibial growth in first filial generation rats, iodine deficiency retarded the chondroncyte proliferation and decreases the thickness of growth plate cartilage in first filial generation rats, and combined selenium and iodine deficiency significantly impair the growth of bone and cartilage in first filial generation rats.