BACKGROUND: Evidence exists that inhibition of matrix metanoproteinase-2(MMP-2) secretion in the proliferating hernangioma tissue by transfection of adenovirus-active MMP-2(Ad-aMMP-2) cDNA would become an important means for treatment of proliferating hemangioma.OBJECTIVE: To investigate the influences of Ad-aMMP-2 cDNA transfection on human proliferating hemangioma growth in nude mice.DESIGN, TIME AND SETTING: A randomized, grouping, and controlled observation was performed in West China Hospital of Sichuan University between August 2003 and September 2004.MATERIALS: Eighteen BALB/c-nu/nu nude mice, weighing approximately 20 g, were included. Cavernous hemangioma specimen pathologically confirmed as proliferating hemangioma was resected from one 52-day-old female child patient.METHODS: The freshly reseoted human proliferating hemangioma specimen was sliced into small pieces with a size of 5 mm×4 mm×3 mm and subcutaneously implanted into the back of 18 nude mice within 1 hour to develop mouse models of hemangioma.Forty-five days after hemangioma implantation, 15 successful hemangioma nude mice were treated by intratumoral administration of adenovirus green fluorescent protein (Ad-GFP1 n = 51 Ad-GFP group), adenovirus-active MMP-2 (n = 5, Ad-aMMP-2 group), or the same amount of phosphate buffered saline (PBS1 n = 51 control group). Intratumoral administration was performed once every other day, for a total of 4 times.MAIN OUTCOME MEASURES: Observation of tumor volume and compadson of tumor necrosis area among 3 groups; detection of GFP expression in nude mouse; gross, hematoxylin-eosin staining, and transmission etectron microscope observation of tumor tissue morphology; determination of MMP-2 cDNA expression and microvascular density by immunohistochemistry; and detection of growth cycle and apoptosis of tumor cells by flow cytometry.RESULTS:①Ad-aMMP-2 could inhibit hemangioma growth in vivo, without marked adverse reactions. Tumor necrosis of different degrees was found in each group, and tumor necrosis area was significantly greater in the Ad-aMMP-2 group than in the control and Ad-GFP groups (P < 0.01). ②Histological sections displayed GFP gene expression in the Ad-GFP group. ③Gross observation results revealed relatively large tumor tissue in the control and Ad-GFP groups and relatively small tumor tissue in the Ad-aMMP-2 group. Hernatoxylin-eosin staining results showed that in the control and Ad-GFP groups, endothelial cells aggregated together in strip-shaped or lump-shaped appearance, and in the Ad-aMMP-2 group, there were many necrotic loci arranging in lamellar-shape appearance. Transmission electron microscope results revealed vascular endothelial cells with normal morphology in the control group and tumor cells with apparent nucleoli in the Ad-GFP group, while in the Ad-aMMP-2 group, some vascular endothelial cells exhibited chromatin pycnosis in the nucleus, forming apoptotic bodies.④ MMP-2 expression and microvascular density were significantly reduced in the Ad-aMMP-2 group than in the Ad-GFP and control groups (P < 0.05). ⑤The percentage of tumor cells in G0/G1 phase was significantly higher (P < 0.05), while the proliferating index was significantly decreased, in the Ad-aMMP-2 group than in the Ad-GFP and control groups. The Ad-aMMP-2 group exhibited higher apoptosis rate of tumor cells (P < 0.05), as well as more markedly increasing apoptosis index, than the control and Ad-GFP groups.CONCLUSION: It is feasible to block human proliferating hemangioma growth by transfeotion of Ad-aMMP-2 cDNA. The included mechanisms are to inhibit vascular endothelial cells to secrete MMP-21 thereby leading to local ischemia.

This study was purposed to investigate the methylation status of id4 gene promoter in patients with chronic myeloid leukemia (CML) and explore the relationship between methylation of the id4 gene and progress of CML. The methylation status of id4 gene in 48 chronic myeloid leukemia patients and 10 healthy individuals was detected by using methylation-specific polymerase chain reaction (MS-PCR). The results showed that id4 gene was unmethylated in bone marrow samples from both healthy individuals and CML patients in chronic phase (CP). The rate of id4 gene methylation in both CML patients in accelerated phase (AP) and blast crisis (BC) was 66%, and was higher than those of CML patients in CP phase. There was significant difference between them (p < 0.05). In one CML patient who received a serial observations, the status of id4 was unmethylated in CP, but it was methylated in AP and BC phase. It is concluded that the id4 gene in CML patients is unmethylated in CP, while it is methylated in AP or BC. The detection of id4 gene methylation status may be useful for monitoring disease advance in CML and may be used as a marker of disease progression in CML.
Adolescent ; Adult ; Aged ; Child ; DNA Methylation ; DNA Primers ; Female ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Male ; Middle Aged ; Promoter Regions, Genetic ; Young Adult

AIMTo indicate the titration end-point of precipitation reaction by measuring the relative intensity of the scattered light in the titrate for use in pharmaceutical analysis.

METHODSA visible light-emitting diode (LED) was used as a light source and a photodiode was used as the optical detector. Light on the detector creates an electric current through the diode. With the addition of the titrant, the titrate became turbid and the intensity of the scattered light in the solution increased gradually. If the precipitation reaction proceeded the completion and the solubility of the precipitate was small enough, the intensity of the scattered light will reach maximum at the stoichiometric point; thus, the titration end-point can be indicated. The accuracy of nephelometric titrimetry was discussed theoretically and the titration of NaCl with AgNO3 was used as a model. To demonstrate the applicability of the new titrimetry in pharmaceutical analysis, phenytoin sodium and procaine hydrochloride were titrated with AgNO3 and NaOH solutions, respectively.

RESULTSWith our new titrator and nephelometric sensor, the accuracy and precision of our new titrimetry can be better than 0.2% under suitable conditions.

CONCLUSIONThis new titrimetry can be used for pharmaceutical analysis.

Phenytoin ; analysis ; Procaine ; analysis ; Titrimetry ; instrumentation ; methods

To sum up the reform of internal performance appraisal system and income allocation system of Shanghai municipal hospitals.The internal performance appraisal index system,evaluation methods and corresponding income allocation system,featuring two breaks,one change and eight elements.The reforms highlight public welfare nature of public hospitals,which is expected to create profound impacts on hospital operation and medical staff behavior.

BACKGROUNDThe diagnosis of myelodysplastic syndrome (MDS), especially hypoplastic MDS, and MDS with low blast counts or normal karyotype may be problematic. This study characterized ID4 gene methylation in patients with MDS and aplastic anemia (AA).

METHODSThe methylation status of ID4 was analyzed by bisulfite sequencing polymerase chain reaction (PCR) and quantitative real-time methylation-specific PCR (MethyLight PCR) in 100 patients with MDS and 31 patients with AA.

RESULTSThe MDS group had a higher ID4 gene methylation positivity rate (22.22%) and higher methylation levels (0.21 [0-3.79]) than the AA group (P < 0.05). Furthermore, there were significant differences between the hypoplastic MDS and AA groups, the MDS with low blast count and the AA groups, and the MDS with normal karyotype and the AA groups. The combination of genetic and epigenetic markers was used in much more patients with MDS (62.5% [35/56]) than the use of genetic markers only (51.79% [29/56]).

CONCLUSIONSThese results showed that the detection of ID4 methylation positivity rates and levels could be a useful biomarker for MDS diagnosis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anemia, Aplastic ; genetics ; Child ; CpG Islands ; genetics ; DNA Methylation ; genetics ; Female ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Young Adult

It is hard to discriminate myelodysplastic syndrome(MDS) from many benign hematological diseases. To identify the methylation status of zo-1 gene in MDS, the methylation specific PCR (MS-PCR) and reverse transcription-PCR (RT-PCR) were applied to detect the MDS cell line MUTZ-1, bone marrow of a healthy donor and an aplastic anemia patient. MS-PCR was also employed to detect the bone marrow of 72 patients with benign hematological diseases, 35 MDS-RA patients, and 20 MDS-like patients. The results showed that MDS cell line MUTZ-1 displayed complete methylation of zo-1 promoter without mRNA expression. Inversely, a patient with benign hematological disease and a donor with normal bone marrow showed complete unmethylation of this gene with unaffected mRNA expression. No zo-1 promoter methylation was detected in patients with benign hematological diseases, while aberrant hypermethylation of zo-1 gene promoter were found in 48.6% (18/37) of MDS-RA patients. The positive rate of zo-1 methylation in MDS-RA patients was higher than that in patients with benign hematological diseases (p < 0.05). Seven suspected MDS patients manifested hypermethylation status of zo-1 gene (7/20), 2 were followed up for 1 year and transformed into MDS. It is concluded that relatively high hypermethylation rate of zo-1 promoter is observed in MDS-RA, and no methylation in patients with benign hematological diseases. Therefore, zo-1 gene hypermethylation may be served as a useful epigenetic marker in the differential diagnosis for MDS.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA Methylation ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; diagnosis ; genetics ; Young Adult ; Zonula Occludens-1 Protein ; genetics ; metabolism

Multiple myeloma (MM) is an incurable heterogeneous disease derived from malignant clonal expansion of plasma cells. The evaluation of prognosis, detection of minimal residual disease (MRD) and treatment of MM are unclear for decades. Recently, Velcade and autotransplantation have been broadly applied to MM patients and achieved better outcomes, but there is yet no effective and universal marker for MRD detection in MM. Both genetic and epigene-tic aberrations play important roles in the pathogenesis and development of cancer. Our preliminary data showed that aberrant promoter methylation of zo-1 and id4 genes was correlated with their gene silencing in several types of hematological malignancies. Therefore, this study was aimed to identify the promoter methylation status of zo-1 and id4 genes in MM and their relationship with the prognosis, MRD and treatment of MM. The methylation status of zo-1 and id4 genes of MM cell lines U266, H929 and IM9 was tested by using MS-PCR; the methylation status of zo-1, id4 gene promoters in bone marrow samples of 20 MM patients and 6 healthy persons was detected by MS-PCR. The results showed that the zo-1, id4 gene in MM cell lines all were methylation positive (complete or partial methylation), the zo-1, id4 gene in samples of 5 healthy persons all were completely unmethylated. The methylation positive rate of zo-1 and id4 genes were 50% and 85% respectively, which were significantly higher than that in normal bone marrow (0%). The coverage rate of zo-1 and id4 gene methylation was 95%. There were no significant differences in the methylation status of both genes among the patients with different heavy chains, different light chains and symptoms. It is concluded that the change of zo-1, id4 genes methylation status occurs in MM patients and has specificity, which may be a new gene marker of MM, methylation analysis of zo-1 and id4 genes may be important for MRD monitoring in patients with MM.
Adult ; Aged ; Bone Marrow ; metabolism ; Cell Line, Tumor ; DNA Methylation ; Female ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; metabolism ; Male ; Membrane Proteins ; genetics ; metabolism ; Middle Aged ; Multiple Myeloma ; genetics ; metabolism ; Neoplasm, Residual ; genetics ; Phosphoproteins ; genetics ; metabolism ; Zonula Occludens-1 Protein

OBJECTIVETo study the methylation status of fragile histidine triad (FHIT) gene promoter in patients with myelodysplastic syndrome (MDS) and its clinical relevance.

METHODSMethylation-specific PCR (MSP) was used to detect FHIT promoter methylation in bone marrow samples from 54 MDS cases.

RESULTSHypermethylation of FHIT promoter was detected in 26 cases (48.1%). Association was not found between FHIT gene hypermethylation and sex, hematologic parameters and chromosomal abnormalities of MDS patients, but found between FHIT gene hypermethylation and age of the MDS cases. Although significant difference was not observed in the frequencies of FHIT gene hypermethylation among patients with refractory anemia/refractory anemia with ringed sideroblasts (RA/RAS) (1/6, 16.7%), refractory anemia/refractory anemia with ringed sideroblasts (RCMD) and refractory cytopenia with multilineage dysplasia with ringed blasts (RCMD-RS) (6/19, 31.6%), refractory anemia with excess blasts-1 (RAEB-1) (7/11, 63.6%), refractory anemia with excess blasts-2 (RAEB-2) (4/7, 57.1%) and refractory anemia with excess blasts in transformation/acute myeloid leukemia (RAEBt/AML) (8/11, 72.7%)(chi-square=8.417, P=0.077), it was observed in patients in early stages (RA/RAS and RCMD) (7/25, 28.0%), advanced stages (RAEB-1 and RAEB-2)(11/18, 61.1%) and RAEBt/AML (8/11, 72.7%) (chi-square=7.938, P=0.019). Furthermore, there was a positive correlation between the frequency of FHIT gene hypermethylation and different IPSS groups (chi-square=10.110, P=0.018).

CONCLUSIONFHIT gene hypermethylation might be one of the molecular events involved in the disease progression of MDS.

Acid Anhydride Hydrolases ; genetics ; Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Base Sequence ; DNA Methylation ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Myelodysplastic Syndromes ; classification ; genetics ; pathology ; Neoplasm Proteins ; genetics ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics

OBJECTIVETo investigate the prevalence of human papillomavirus (HPV) and the HPV genotype distribution in invasive squamous cell carcinoma of the uterine cervix in the Mongolian women in Inner Mongolia autonomy region.

METHODSThe prevalence data of HPV in our department were retrospectively reviewed. INNO-LiPA genotyping technique was used to detect HPV genotypes in the reserved carcinoma tissue specimens.

RESULTSTotally 63 tissue specimens were collected and detected. The prevalence of HPV was 93.7%. The positive rates of HPV among different clinical staging and different pathological grading were not significantly different (P >0.05). The prevalence of HPV16 was not significantly different among different age groups (P>0.05). HPV16 (69.8%), HPV18 (4.8%), HPV31 (4.8%), HPV39 (4.8%), and HPV52 (3.2%) were the 5 dominating HPV genotypes in all cases.

CONCLUSIONSHPV infection is closely correlated with invasive squamous cell carcinoma of the uterine cervix in Mongolia women. HPV16 is the most important genotype in invasive squamous cell carcinoma of the uterine cervix, followed by HPV18, 31, and 39. HPV infection dose not affect the progression and differentiation of invasive squamous cell carcinoma of the uterine cervix.

Adult ; Aged ; Asian Continental Ancestry Group ; Carcinoma, Squamous Cell ; virology ; Female ; Genotype ; Humans ; In Vitro Techniques ; Middle Aged ; Papillomaviridae ; classification ; genetics ; Papillomavirus Infections ; genetics ; virology ; Polymerase Chain Reaction ; Uterine Cervical Neoplasms ; virology

HIV/AIDS patients in high prevalence areas with different routes of infection (sexually transmitted 878 cases, 527 cases of intravenous drug user, paid blood donor 652 cases) were choosen for traditional Chinese medicine (TCM) syndrome investigation for one-year clinical follow-up. This paper primarily concluded the nature, location and pathogenesis of AIDS diseases. Deficiency of Yang and Yin, combining deficiency of Qi are the basic deficiency syndromes, while stagnation of dampness, toxic fire are the excess syndromes; the disease location of HIV infector is spleen, main syndrome is deficiency of spleen Qi; the disease location of AIDS patient is kidney, main syndrome is deficiency of spleen and kidney Yang. The pathogenic development tendency is from deficiency of Qi to combining stagnation of dampness and toxic fire, finally to deficiency of Qi and Yin, deficiency of Yang.
Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; HIV Infections ; diagnosis ; etiology ; transmission ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Young Adult