Child ; Child, Preschool ; Female ; Gastrointestinal Hemorrhage ; diagnosis ; Gastrointestinal Tract ; diagnostic imaging ; Humans ; Infant ; Male ; Radionuclide Imaging ; Sodium Pertechnetate Tc 99m

OBJECTIVETo explore the mechanism of miR-124 inhibiting the proliferative activity of prostate cancer PC3 cells.

METHODSLuciferase reporter gene assay was used to examine the specific binding ability of miR-124 to PKM2 mRNA 3'-UTR. After miR-124 was transfected mimic to PC3 cells, the expression levels of PKM2 mRNA and protein were detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, respectively. The effects of miR-124 mimic and PKM2 siRNA on the proliferative activity of the PC3 cells were determined by MTT assay.

RESULTSThe expressions of PKM2 mRNA and protein were upregulated (5.12 +/- 0.35) times and (4.05 +/- 0.20) times respectively in the PC3 cells as compared with those in the RWPE-1 cells (P < 0.05). Luciferase reporter gene assay demonstrated that miR-124 targeted PKM2 3'-UTR. At 24 hours after transfection with miR-124 mimic, the PKM2 protein expression in the PC3 cells was downregulated (0.16 +/- 0.04) times (P < 0.05), while the PKM2 mRNA level was not changed significantly (P > 0.05), as compared with the control group. MTT assay showed that both miRNA-124 mimic and PKM2 siRNA could inhibit the proliferation of the PC3 cells, but the former exhibited a greater inhibitory effect than the latter. After transfection with miR-124 mimic and PKM2 siRNA, the cell growth rates were (66.20 +/- 5.10)% vs (82.10 +/- 6.35)% at 24 hours (P < 0.05) and (49.34 +/- 2.37)% vs (70.10 +/- 5.80)% at 48 hours (P < 0.05).

CONCLUSIONmiR-124 can suppress the proliferation of PC3 cells by regulating the PKM2 gene.

Carrier Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; genetics ; Humans ; Male ; Membrane Proteins ; genetics ; metabolism ; MicroRNAs ; genetics ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; Thyroid Hormones ; genetics ; metabolism ; Transfection

OBJECTIVETo study the effect of silencing pyruvate kinase M2 (PKM2) on gambogic acid (GA)-induced apoptosis of human prostate cancer PC3 cells.

METHODSThree specific PKM2 siRNAs and one negative control siRNA (si-NC) were transfected into PC3 cells. The silencing effect of PKM2 siRNAs was determined by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, and the effects of PKM2 siRNA on the vitality and apoptosis of GA-stimulated PC3 cells detected by MTT and AO/EB double staining, respectively. The mRNA and protein levels of c-myc and cyclin D1 were analyzed by qRT-PCR and Western blot, respectively.

RESULTSAll the 3 PKM2 siRNAs effectively reduced the mRNA and protein expressions of PKM2, and PKM2 siRNA-1 exhibited the strongest silencing effect. At 24 h after transfection, the expression levels of PKM2 mRNA and protein were reduced by 70% and 85%, respectively (P < 0.05). Twenty-four hours of treatment with GA (0.5 micromol/L) following transfection with PKM2 siRNA-1 inhibited the vitality of the PC3 cells by 68%, increased their apoptosis, and significantly down-regulated the mRNA and protein levels of c-myc (50% and 35%) and cyclin D1 (60% and 20%) (P < 0.05).

CONCLUSIONInhibition of PKM2 sensitized PC3 cells to GA-induced apoptosis, suggesting that PKM2 may be a potential therapeutic target for sensitizing human prostate cancer to GA.

Apoptosis ; drug effects ; Carrier Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Humans ; Male ; Membrane Proteins ; genetics ; metabolism ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; RNA Interference ; RNA, Small Interfering ; Thyroid Hormones ; genetics ; metabolism ; Xanthones ; pharmacology

OBJECTIVETo investigate a method for the repair of tissue defect.

METHODSAllogenic acellular dermal matrixes (ADM) were implanted to full-thickness skin defects made on the dorsa of rats. Two weeks later, autologous suspended epidermal cells were transplanted on to the surface of vascularized ADM. Respectively, neoepidermis was macroscopically observed 2, 3, 5 weeks after grafting, and samples were taken to make routine paraffin sections for microscopical examination, and immunohistochemical staining for type IV collagen was also performed.

RESULTSThe vascularized ADM could support proliferation and differentiation of epidermal cells, and also could promote the formation of dermal-epidermal junction. Suspended epidermal cells in an artificial culture system in vivo could develop into mature epidermis. The reconstructed skin not only looked like the normal one in appearance in which hair was removed, but also revealed a better function.

CONCLUSIONSFull-thickness skin defect can be repaired by transplanting autologous epidermal cell suspension on to vascularized ADM.

Animals ; Cell Transplantation ; Dermis ; cytology ; Epidermis ; cytology ; Extracellular Matrix ; Rats ; Rats, Wistar ; Skin ; injuries ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Suspensions ; Tissue Engineering ; Transplantation, Heterologous ; Wound Healing

OBJECTIVETo investigate the expression of vascular endothelial growth factor-C (VEGF-C) and cyclooxygenase-2 (COX-2) proteins, and their relationship with biological behaviors of non-small-cell lung carcinoma (NSCLC).

METHODSImmunohistochemical staining was used to detect the expression of VEGF-C and COX-2 proteins in 77 cases of NSCLC. The relationship was analyzed between the expression of VEGF-C, COX-2 and lymphatic vessel density (LVD), tumor size, histological type, differentiation, lymph node metastasis, clinical recurrence and survival time of the patients.

RESULTSOut of 77 cases of NSCLC, 45 cases and 29 cases showed positive expression of VEGF-C and COX-2 proteins, respectively. The expression rates of VEGF-C and COX-2 protein were 58.4% and 37.7%, respectively. The expression of VEGF-C protein was correlated negatively with the degree of differentiation of NSCLC (P < 0.05). The expression of VEGF-C was positively correlated with lymph node metastasis, LVD and tumor size (P < 0.01). The survival time of the patients was negatively correlated with the expression of VEGF-C (P < 0.01). The expression of COX-2 was positively correlated with LVD (P < 0.01). The survival time of the patients was negatively correlated with the expression of COX-2 (P < 0.05).

CONCLUSIONThe expression of VEGF-C and COX-2 proteins are closely correlated with the biological behaviors of NSCLC, especially VEGF-C protein. Its high expression suggests probable lymph node metastasis and poor prognosis.

Adult ; Aged ; Biomarkers, Tumor ; biosynthesis ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cyclooxygenase 2 ; biosynthesis ; Female ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Lymphangiogenesis ; Lymphatic Metastasis ; Male ; Middle Aged ; Prognosis ; Vascular Endothelial Growth Factor C ; biosynthesis

OBJECTIVETo study the clinicopathologic features, immunohistochemical findings, EBV and c-myc gene status of intra-abdominal non-Hodgkin B-cell lymphoma occurring in children.

METHODSSeventy-four cases of pediatric intra-abdominal non-Hodgkin B-cell lymphoma were retrieved from the archival file. The cases were classified according to the 2008 WHO classification. Tissue microarray including tumor tissues from all the 74 cases was produced. Immunohistochemical study (SP method) for CD20, CD3, CD79a, CD10, bcl-6, MUM1, bcl-2, CD43, CD38 and Ki-67 was performed. In-situ hybridization for Epstein-Barr virus-encoded RNA (EBER) and fluorescence in-situ hybridization for c-myc gene were also carried out.

RESULTSAmongst the 74 cases studied, 65 of them (87.8%) were Burkitt lymphoma (BL), 4 cases (5.4%) were diffuse large B-cell lymphoma (DLBCL) and the remaining 5 cases (6.8%) showed features in-between DLBCL and BL (DLBCL/BL). The patients often presented with abdominal pain, abdominal masses, ileus and intussusception. The ileocecal bowel wall and mesenteric lymph nodes were commonly involved. The lymphoma cells were of high histologic grade and suggested an aggressive clinical behavior. The staining for CD20 and CD79a were positive in all of the cases, while CD3 was negative. The positive rates of CD10, bcl-6, bcl-2, MUM1, CD43, CD38 and EBER in BL were 96.9% (63 cases), 95.4% (62 cases), 0 (0 case), 23.1% (15 cases), 70.8% (46 cases), 96.9% (63 cases) and 41.5% (27 cases), respectively. Fifty-four cases carried translocation of c-myc gene. As for DLBCL, the positive cases of CD10, bcl-6, bcl-2, MUM1, CD43, CD38 and EBER were 3 cases, 2 cases, 3 cases, 2 cases, 2 cases, 2 cases and 0 case, respectively. One of these cases showed c-myc gene translocation. Amongst the 4 cases of DLBCL, 2 of them belonged to germinal center B-cell-like subtype, while the remaining 2 cases were of non-germinal center B-cell-like subtype. The expression rates of CD10, bcl-6, bcl-2, MUM1, CD43, CD38 and EBER in DLBCL/BL were 5/5, 4/5, 0, 3/5, 5/5, 3/5 and 0, respectively. Three of the cases were positive for c-myc gene translocation.

CONCLUSIONSThe majority of pediatric intra-abdominal non-Hodgkin B-cell lymphoma belonged to BL. They have characteristic clinical presentation and sites of predilection and are often associated with an aggressive clinical behavior. Thorough morphologic assessment, immunohistochemistry and in-situ hybridization play an important role in subtyping this group of lymphoid malignancy.

Antigens, CD20 ; metabolism ; Burkitt Lymphoma ; genetics ; metabolism ; pathology ; CD79 Antigens ; metabolism ; Child ; Child, Preschool ; Female ; Genes, myc ; Humans ; Intestinal Neoplasms ; genetics ; metabolism ; pathology ; Lymphoma, B-Cell ; genetics ; metabolism ; pathology ; Lymphoma, Large B-Cell, Diffuse ; genetics ; metabolism ; pathology ; Male ; Neprilysin ; metabolism ; RNA, Viral ; metabolism ; Translocation, Genetic

OBJECTIVETo assess the diagnosis and treatment of invasive lung aspergillosis after liver transplantation.

METHODSRoutine sputum culture was performed. Itraconazole and fluconazole were used to prevent fungal infection prophylactically. Amphyotericin B was only used on aspergillosis. In 54 patients receiving, liver transplantation, 3 patients with lung aspergillosis were reviewed.

RESULTSOf the 3 patients 2 died and 1 recovered.

CONCLUSIONSOver-immunosuppression is a main risk factor for aspergillosis. Amphotericin B is still the best choice for the treatment of aspergillosis and its gradual, interrupted, low concentration administration, cooperated with itraconazole can ease the side effects.

Adult ; Aspergillosis ; diagnosis ; drug therapy ; etiology ; Female ; Humans ; Liver Transplantation ; adverse effects ; Lung Diseases, Fungal ; diagnosis ; drug therapy ; etiology ; Male ; Middle Aged

OBJECTIVETo investigate the knowledge and attitudes of healthcare professionals (doctors, nurses and administrators) to adverse drug reactions (ADR) in Wuhan city and to identify the reasons for under-reporting.

METHODSStructured interviews were carried out in Wuhan, Hubei province. Questionnaire survey to approximately 15% of the medical practitioners selected from 16 hospitals, was conducted during the period from February to March 2003.

RESULTSOnly 2.7% of the interviewees knew the definition of adverse drug reactions. 61.7% of the doctors, 62.7% of the nurses and 61.1% of the administrators had ever encountered an ADR during their practices, but did not report to the national monitoring center or other centers. The major reasons for not reporting included: ignorant about the requirement and the reporting process of ADR (71.4%); address of the reporting agency and Forms unavailable (67.9%, 60.4%); unaware of the existence of a national ADR reporting system (52.2%); needless to report as the ADR being too well known (44.1%). They mainly reported an ADR to the hospital pharmacy or other departments, or to the pharmaceutical administration. Education, training and developing new institutions were ways to improve the reporting system.

CONCLUSIONSOur results showed that healthcare professionals had little knowledge on the basic ADR knowledge. The main reasons for underreporting were related to factors on reporting process, address of related centers and unavailable of the Forms. Education and training to doctors and nurses to enhance the awareness of administrators were the ways to improve the reporting system.

Adverse Drug Reaction Reporting Systems ; Attitude of Health Personnel ; China ; Drug-Related Side Effects and Adverse Reactions ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Practice Patterns, Physicians' ; Surveys and Questionnaires

BACKGROUNDA voluntary procedure for reporting adverse drug reactions (ADRs) was formally put in place in 1989. However, only a small proportion of ADR reports are actually forwarded to the national monitoring center. To identify the reasons for underreporting, the authors investigated the awareness and attitudes of healthcare professionals (doctors, nurses, and administrators) toward the ADR system in China. In addition, the authors sought to formulate approaches to improve the current ADR reporting system.

METHODSStructured interviews were carried out in 16 hospitals selected from 27 municipal hospitals in Wuhan, Hubei Province, China. A questionnaire survey of a stratified random sample of approximately 15% of healthcare professionals in each selected hospital was conducted during February to March 2003.

RESULTSThe response rate of this survey was 85%. One thousand six hundred and fifty-three questionnaires were used in the final analysis. Only 2.7% of the healthcare professionals had a correct understanding to the definition of ADR. Eighty-nine point two percent of the healthcare professionals had encountered ADRs. Ninety-four percent of them were aware of the need to report these to the ADR monitoring center. However, only 28.5% of doctors, 22.8% of nurses, and 29.7% of administrators actually submitted a report. For the most part, they reported ADRs to the hospital pharmacy (66.0%), to other departments in the hospital (72.5%), and to the pharmaceutical industry (23.0%), rather than to the national monitoring center (2.9%) or regional monitoring center (9.5%). Severe or rare ADRs and ADRs to new products were generally perceived to be significant enough to report. Sixty-two point one percent of the healthcare professionals had encountered ADRs, yet not reported them to anybody. The major reasons for not reporting included no knowledge of the reporting procedure (71.4%), unavailability of the reporting center mailing address (67.9%), unavailability of the ADR report form (60.4%), lack of knowledge of the existence of a national ADR reporting system (52.2%), and belief that the ADR in question was already well known (44.1%).

CONCLUSIONSHealthcare professionals in Wuhan, China have little basic knowledge of ADR and of the voluntary reporting system. The main reasons for underreporting were lack of basic knowledge about ADRs and the voluntary reporting procedure. Education and training of healthcare professionals is needed to improve the current ADR reporting system.

Adverse Drug Reaction Reporting Systems ; trends ; Attitude of Health Personnel ; China ; Health Knowledge, Attitudes, Practice ; Hospital Administrators ; Humans ; Interviews as Topic ; Nurses ; Physicians ; Surveys and Questionnaires

We designed the specific primers and TaqMan probes targeting cytochrome b genes of mitochondrial DNA from bovine, goat and sheep. We used different fluorescents to label the probes. After optimization of reaction conditions, we set up a multiplex fluorescent real-time PCR method to detect bovine, goat and sheep derived materials, simultaneously. We finished the detection tests of 17 kinds of animal DNA and 200 DNA samples from different sources with the developed method and the National Standard GB/T 20190Y-2006 routine PCR method. The coincidence rate of these two methods was 100%. Without electrophoresis or restriction digestion, the developed method could reduce the test time to one third as routine PCR and identify three kinds of animal derived materials including bovine, goat and sheep in one reaction. The developed method was approximately 10 times more sensitive than routine PCR, and was applicable to identifications of bovine, goat and sheep derived materials in feed stuff, meat, milk, pelt and grease, etc. The study showed that the developed real-time PCR method is a rapid, sensitive and efficacious detection assay for bovine, goat and sheep derived materials in animal products.
Animal Feed ; analysis ; Animals ; Cattle ; Cytochromes b ; genetics ; DNA Primers ; DNA, Mitochondrial ; analysis ; genetics ; Food Contamination ; analysis ; Genes, Mitochondrial ; Goats ; Meat ; analysis ; Mitochondria, Muscle ; enzymology ; genetics ; Polymerase Chain Reaction ; methods ; Sheep