Objective • To investigate whether methylene blue-mediated photodynamic therapy (MB-PDT) can induce the apoptosis of macrophages in periodontitis, and simultaneously reduce the bone absorption. Methods • For in vitro treatments, cells were divided into three experimental groups: control group, no treatment; MB group, methylene blue treatment; MB-PDT group, MB and laser irradiation treatment. Then apoptosis and apoptosis related genes were detected in each group. For in vivo treatments, periodontal disease in SD rats was orthodontic ligature and periodontal pathogen induced at the first maxillary molar. After 6 weeks, the ligature was removed and all animals received scaling and root planning(SRP) and were divided according to the following treatments: SRP group, saline solution; MB group, phenothiazinium dye; and MB-PDT group, MB and laser irradiation, once a week. All animals in each treatment were killed after 3 weeks. Immunofluorescence and Micro CT analyses were used to detect the apoptotic macrophages and alveolar bone resorption in periodontal tissues of rats in each group. Results • In vitro experiments showed that the combination of 10 μmol/L MB and 40 J/cm2 light dose could kill more than 50% macrophages, and the apoptosis of macrophages induced by MB-PDT in this mode was most obvious. Meanwhile, MB-PDT increased the expression ratio of proapoptotic gene Bax /antiapoptotic gene Bcl-2 in macrophages. In vivo experiments showed that the macrophages infiltrated in tissues with periodontitis were apoptotic after MB-PDT treatment, while no obvious apoptosis of macrophages infiltrated in the periodontal tissues of SRP and MB rats was found. Micro CT analysis showed that the alveolar bone resorption in MB-PDT rats was less than that in SRP and MB rats (P<0.05), and there was no significant difference between SRP rats and MB rats. Conclusion • MB-PDT can induce the apoptosis of hyperproliferating macrophages in periodontitis and reduce the bone absorption. Compared with SRP, MB-PDT is an effective adjunctive treatment of periodontitis.

To investigate the biological characteristics of the variant translocation der ins (17;15) in a patient with acute promyelocytic leukemia (APL), the conventional G-banding technique, interphase fluorescence in situ hybridization (int-FISH), RT-PCR, gene scanning, gene sequence and flow cytometry were performed. The results indicated that the variant translocation der ins (17, 15) observed by G banding technique was a rare type, the int-FISH assay by using dual-color pml/raralpha fusion probes confirmed the cytogenetic findings. The detection results of other molecular methods demonstrated the existence of the whole pml/raralpha fusion gene, while this case had insertion variant translocation. This patient got complete remission by using combined chemotherapy, and survives with continuous complete remission during following up for 1 year. In conclusion, the variant translocation der ins (17; 15) is rare type in APL, its incidence is lower, several signal types in detection of int-FISH were observed and the combination chemotherapy for this patient showed more obvious efficacy.
Chromosome Banding ; Chromosomes, Human, Pair 15 ; Chromosomes, Human, Pair 17 ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Interphase ; genetics ; Leukemia, Promyelocytic, Acute ; genetics ; Male ; Translocation, Genetic ; Young Adult

This study was purposed to analyze the characteristics of morphology, immunology, cytogenetic and molecular biology of leukemia cells in 12 AML patients with Ph(+) and their correlation with survival of patients. 12 patients with Ph(+) AML were diagnosed according to diagnostic criteria of WHO and existence of t(9;22) (q34;q11) or t(9;22) abnormality, meanwhile no evidence of CML chronic phase was observed. The results showed that 8 out of 12 cases were confirmedly diagnosed to be AML by morphologic and immunophenotypic examination, 4 cases were diagnosed as myeloid and B lymphocytic mixed acute leukemia. The Ph chromosome was detected in 10 cases by chromosome analysis at the first time of diagnosis, and some of the cases had coexistence of complex chromosome and/or normal karyotype. BCR-ABL transcript was detected in all 12 cases, including 7 cases with b3a2, 1 case with b2a2, 1 case with b2a2 variants, 2 cases with e1a2 and 1 case with e18a2. The 12 cases all got complete remission after chemotherapy and/or gleevec treatment, out of them 3 cases received chemotherapy and gleevec treatment, but 2 cases died; 9 cases received allogeneic hematopoietic stem-cell transplantation (allo-HSCT), 1 case died from relapse, among them 1 case died from transplant complications. The median survival was 24 (8 - 80) months, the overall survival of 3 years was (51.4 ± 17.7)%. It is concluded that the Ph(+) AML is a acute myelogenous leukemia with poor prognosis, but long-term survival may be achieved with HSCT as quick as after complete remission from gleevec and chemotherapy treatment. Meanwhile, the detection of BCR-ABL gene and it variants may be give more opportunity for diagnose and treatment, which can be used as routine screening for newly diagnosed leukemia.
Adult ; Child ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; diagnosis ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; Prognosis

AIMTo study the metabolite of stilbene glucoside in the Chinese traditional medicine Polygonum multiflornm in mice and elucidate its chemical structure by liquid chromatography tandem-mass spectrometry.

METHODSThe stilbene glucoside was injected into the tail vein of mice. Blood samples were collected from artery in the eyepit. The methanol-protein-precipitated plasma sample was introduced into the liquid chromatography-tandem mass spectrometer directly. The analytical column was C18 column (250 mm x 4.6 mm ID, 5 microns). The mobile phase consisted of acetonitrile-methanol-water-formic acid (15:18:66:1) for ES+, acetonitrile-methanol-water (15:18:67) for ES-. The UV detection wavelength was set at 320 nm. The mass ion source type was ESI. HV capillary was 3 kV. The dry gas was nitrogen gas and the flow rate was set at 318 L.h-1. The ion source temperature was 150 degrees C.

RESULTSThe stilbene glucoside and its metabolite were separated completely under the chromatography condition. The ions at m/z 600 and m/z 605 were detected under positive ion polarity while the ions at m/z 581 and m/z 402 were detected under negative ion polarity.

CONCLUSIONIt was proposed that the metabolite of stilbene glucoside injected in vein was its glucuronide conjugate.

Animals ; Chromatography, Liquid ; methods ; Female ; Glucosides ; blood ; isolation & purification ; metabolism ; Male ; Mice ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Stilbenes ; blood ; chemistry ; isolation & purification ; metabolism

OBJECTIVETo look for the evidences of motilin receptor expression on interstitial cells of Cajal (ICC) of the rabbit.

METHODSmooth muscle segments with ICC were isolated from the small intestine of 10-day old rabbits. The tissue segments equilibrated in Ca(2+)-free Hanks' solution were dispersed with an enzyme solution containing collagenase type II and then Ficoll density centrifugation was used to dissociate ICC. The cells were suspended and cultured in the M199 medium. The c-kit antibody was applied to distinguish the cultured ICC. The motilin receptor was identified by immunocytochemical assay with GPR38 antibody, c-kit antibody and hoechst 33342 combined to label ICC. Cells cultured for a few days were sorted for ICC with c-kit stained green fluorescent through flow cytometry. The total RNA and proteins extracted from the sorted ICC were respectively used to verify motilin receptor on the ICC by reverse-transcriptase polymerase chain reaction (RT-PCR) and Western blotting.

RESULTWe had successfully dissociated and cultured ICC of rabbit small intestine in vitro. Fluorescent staining with c-kit antibody confirmed that the culture ICC was successful. Triple-labeled immunofluorescent staining had detected the motilin receptor on membrane of ICC. Flow cytometry analysis showed that the ratio of c-kit positive cell in the cultured cells was 64.3%. The number of sorted ICC was 6.7 × 10(5) and 5.6 × 10(6). The results of RT-PCR and Western blot confirmed that the ICC had motilin receptor expression.

CONCLUSIONOur study demonstrated presence of motilin receptor on ICC of the rabbit. The present results may suggest that ICC play an important role in gastrointestinal movement induced by motilin.

Animals ; Cells, Cultured ; Interstitial Cells of Cajal ; metabolism ; Intestine, Small ; cytology ; Rabbits ; Receptors, Gastrointestinal Hormone ; metabolism ; Receptors, Neuropeptide ; metabolism

The purpose of this study was to investigate whether pretransplant infusion of reactive natural killer cells (NK cells) from donor or recipient can reduce graft-versus-host disease (GVHD) and enhance engraftment in bone marrow transplantation (BMT). Recipient BALB/c mice were divided into 4 groups after received 6.5 Gy total-body irradiation (TBI): control group 1 was treated with nothing, control group 2 received BMT alone, experiment group 1 received BMT and autoreactive NK cells, experiment group 2 received BMT and alloreactive NK cells. Life span, clinical and pathologic changes of GVHD and chimerism rate of each group were evaluated. The results showed that all mice were survival in control group 1. The life span was shorter in experiment group 1 than that in control group 2 (P < 0.05) and longer in experiment group 2 than that in control group 2 (P < 0.01). GVHD was higher in score of experiment group 1 than in control group 2 (P < 0.05) but lower in experiment group 2 than that in control group 2 (P < 0.01). The donor chimerism rate in both two experiment groups were higher than that in control group 2 (P < 0.05), however, the donor chimerism rate was higher in experiment group 2 than that in experiment group 1 (P < 0.01). It is concluded that pretransplant infusion of alloreactive donor NK cells can prolong life span, reduce the degree of GVHD and enhance engraftment. But autoreactive recipient NK cells can shorten life span, aggravate the degree of GVHD and also enhance engraftment, which is weaker than that using alloreactive donor NK cells.
Animals ; Bone Marrow Transplantation ; adverse effects ; Graft Survival ; immunology ; Graft vs Host Disease ; prevention & control ; Killer Cells, Natural ; transplantation ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Whole-Body Irradiation

Comparative analysis of variable region ORF14/15 genes of white spot syndrome virus (WSSV) genome in Guangxi Penaeus vannamei (P. vannamei) could provide useful information for the evaluation of genetic diversity and genetic evolutionary relationship among WSSV isolates from Guangxi, China and other places. Based on geographical and temporal considerations, 40 WSSV-positive P. vannamei samples were collected during the period between May 2010 and July 2013 from Beihai, Qinzhou, and Fangchenggang, which were the main P. vannamei production areas in Guangxi, and the variable region ORF14/15 genes of the WSSV genome from all infected samples were amplified by PCR and then subjected to cloning and sequence analysis. Pairwise and multiple alignment analysis was then conducted to evaluate the degree of genetic divergence between different strains. The variable region ORF14/15 genes from 25 of 40 WSSV positive samples were successfully cloned and sequenced; among the ORF14/15 genes of 25 WSSV-positive strains, 22 was 619 bp in length and 3 was 620 bp. All the 25 Guangxi strains carried a 5949-bp deletion in the ORF14/15 region relative to TH-96-II, which has the longest nucleotide sequence in this region; the deletion of Guangxi strains occurred in the middle region of ORF14/15 gene, with only 190 bp and 429 bp/ 430 bp at 5' and 3' ends, respectively, which were coincident with WSSV-IN-05-I in deletion length and position. Sixteen of 25 Guangxi strains had completely identical nucleotide sequences in the variable re gion, and the homology between other strains also exceeded 97.9%. There were single nucleotide substi tution, deletion, and insertion in the ORF14/15 region of Guangxi strains compared with other strains in GenBank. In the phylogenetic tree based on WSSV variable region ORF14/15, the Guangxi strains were closely related and formed a separate branch with Indian strain IN-05-I, but far from other strains in GenBank. The ORF14/15 gene of WSSV isolates in cultured P. vannamei in Guangxi has a large deletion in the middle of the variable region, and the Guangxi WSSV strains show no significant spatio-temporal differences; the Guangxi strains are closer in genetics to Indian strain IN-05-I than other strains in GenBank.
Animals ; China ; Cloning, Molecular ; Evolution, Molecular ; Genome, Viral ; genetics ; Genomics ; Penaeidae ; virology ; Phylogeny ; White spot syndrome virus 1 ; genetics

OBJECTIVETo study the clinicopathologic features, immunophenotype and ultrastructural features of sinonasal inflammatory myofibroblastic tumors (IMT).

METHODSThe clinical and histologic features of 5 cases of sinonasal IMT were reviewed. Immunohistochemical study for vimentin, MSA, SMA, calponin, h-caldesmon, desmin, ALK, fibronectin, CK, S-100 and Ki-67 was carried out. Ultrastructural examination was also performed in two of the cases.

RESULTSThe patients age ranged from 28 to 62 years (mean = 43 years). The male-to-female ratio was 2:3. The clinical presentation included nasal obstruction, nasal discharge, nasal bleeding, facial pain, facial swelling, toothache and tear overflow. All of the 5 patients suffered from disease relapses; and 4 of them had recurrences for more than 5 times. One patient had lymph node metastasis and 3 patients died of the disease. Histologically, the tumor cells were arranged in interlacing fascicles and sometimes haphazard in fashion. They were spindly in shape, cytoplasm eosinophilic with mild nuclear atypia and a low mitotic activity. The intervening stroma was myxoid in appearance accompanied by lymphocyte and plasma cell infiltration, abundant blood vessels and focal collagenized areas. In 3 of the recurrent cases, the tumor cells displayed increased nuclear atypia and mitotic activity (average about 5 to 6 per 10 high-power fields), accompanied by patchy necrosis, less inflammatory cell infiltration and focal sarcomatous changes. Immunohistochemical study showed that the tumor cells were diffusely positive for vimentin. SMA, MSA, calponin and fibronectin were variably expressed. Desmin was weakly positive in 1 case. The staining for h-caldesmon, ALK, S-100 and CK was negative. The Ki-67 proliferation index increased with tumor recurrences. Electron microscopy revealed abundant rough endoplasmic reticulum and dense body formation in the cytoplasm. There were an increased amount of collagen fibers in the stroma.

CONCLUSIONSIMT rarely occurs in nasal cavity and paranasal sinuses. The tumor is prone to local invasion and recurrences, with subsequent progression to frank malignancy and distant metastasis, resulting in high mortality and poor prognosis. Complete surgical resection remains the main modality of treatment.

Actins ; metabolism ; Adult ; Calcium-Binding Proteins ; metabolism ; Diagnosis, Differential ; Female ; Fibrosarcoma ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Lymphatic Metastasis ; Male ; Microfilament Proteins ; metabolism ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasms, Muscle Tissue ; metabolism ; pathology ; surgery ; ultrastructure ; Neurofibromatoses ; pathology ; Paranasal Sinus Neoplasms ; metabolism ; pathology ; surgery ; ultrastructure ; Vimentin ; metabolism

OBJECTIVETo investigate the application of auditory brainstem response (ABR) and 40 Hz auditory event related potential (40 Hz AERP) to the diagnosis of occupational noise-induced hearing impairment and to provide the evidence for diagnosis of occupational deafness.

METHODSPure tone audiometry, ABR and 40 Hz AERP were performed in 54 workers occupationally exposed to noise. The thresholds of higher frequency band, 3 kHz and 4 kHz were compared with the threshold of ABR. The thresholds of auditory frequency ban and 0.5 kHz were compared with the threshold of 40 Hz AERP.

RESULTSA better correlation was found between thresholds of ABR and higher frequency pure tone audiometry. There was a significant difference of thresholds between 40 kHz AERP and pure tone audiometry. The correction values of thresholds between 40 kHz AERP and pure tone audiometry in the light noise-induced hearing impairment group and the moderate noise-induced hearing impairment group were (16.43 ± 1.08) and (11.80 ± 1.12) dBn HL, respectively.

CONCLUSIONIn diagnosis of occupational noise-induced hearing impairment, the threshold of ABR can be used to estimate the hearing threshold of pure noise higher frequency. Because there is the significant difference of the thresholds between pure tone audiometry and 40 Hz AERP, the response threshold can not be served as the audiometry threshold, and the behavioral hearing thresholds can only be obtained by adjusting the response threshold with respective correction value.

Adult ; Audiometry, Evoked Response ; Audiometry, Pure-Tone ; Auditory Threshold ; Evoked Potentials, Auditory ; Evoked Potentials, Auditory, Brain Stem ; Hearing Loss, Noise-Induced ; diagnosis ; physiopathology ; Humans ; Male ; Middle Aged ; Noise, Occupational ; Young Adult

Background There have been an abundance of literature on the analysis of the mechanical characteristic of the sclera at the entire seleral level in high myopia.However,some recent studies on high myopia are focused on the mechanical changes of the sclera on a cellular level. Objective This experiment was purposed to study how transforming growth factor-β2(TGF-β2)affected sclerotic desmocytes and the mechanical behaviors of scleral fibroblasts in the posterior part of the eyes in guinea pigs with experimental myopia. Methods Induced myopic animal models were established by wearing-10.00 D concave lens for 30 days in lateral eyes of 2-week-old guinea pigs.The fellow eyes were used as control group.Another 5 matched animals served as normal controls.The scleral fibroblasts of each group were purified with the tissue explant method and passaged for 2 generations in vitro.Cultured cells were identified by immunochemistry with vimentin,desmin,keratin and S-100 antibodies.Different concentrations of TGF-β2(0,1,10,100mg/L)were added into serum-free DMEM for 24 hours,and the viscoelastic properties of scleral fibroblasts were measured by micropipette aspiration technique. Results Compared with the fellow,eyes and normal control eyes,the equilibrium modulus and apparent viscosity in model eyes were significantly higher(P＜0.05).After treatment of TGF-β2,the equilibrium modulus and apparent viscosity in the model group and fellow eyes were positively correlated with the concentrations of TGF-β2(r=0.743,r=0.533,r=0.654,r=0.576,P＜0.05).Following the addition of 1 mg/L TGF-β2 and 10 mg/L TGF-β2,the equilibrium modulus and apparent viscosity of scleral fibroblasts were significantly reduced in model eyes compared with fellow eyes(P＜0.05).No significant difference was found in the equilibrium modulus and apparent viscosity of scleral fibroblasts between model eyes and fellow eyes after treatment with 100 mg/L TGF-β2(P＞0.05). Conclusion TGF-β2 car increase the mechanic indexes in a concentration.dependent manner.1 mg/L,10 mg/L TGF-β2 can lower the equilibrium modulus and apparent viscosity of scleral fibroblasts of normal eye and thus cause more changes in the mechanical behavior of scleral fibroblasts.