AIM: To investigate the mechanism of AngRem104-mediated regulation of fibronectin gene in human mesangial cells. METHODS: A series of deleted FN promoter sequences was constructed, which fuse to a luciferase reporter gene, and then the potential active regions that respond to AngRem104 in the upstream regulatory sequence of human FN gene was screened by detecting the luciferase activity of promoter-reporter gene. RESULTS: The detection of relative luciferase activity revealed that there was no significant differences when the HMC were transfected with FN122-reporter gene together with sense AngRem104 construct, but the luciferase activity significantly increased when transfection of FN507-reporter gene construct together with sense AngRem104 construct. However, no increase in luciferase activity was observed when transfection of FN1280-reporter gene construct together with sense AngRem104 construct. The potential regulatory region responds to AngRem104 is in the upstream sequence (-122 to -507) of human FN gene. CONCLUSION: Our preliminary study provided the evidence that AngRem104 may mediate the transcription of the FN gene via the activation of its promoter.

To develop a cell-penetrating chimeric apoptotic peptide AVPI-LMWP/DNA co-delivery system for cancer therapy, we prepared the AVPI-LMWP/pTRAIL self-assembled complexes containing a therapeutic combination of peptide drug AVPI and DNA drug TRAIL. The chimeric apoptotic peptide AVPI-LMWP was synthesized using the standard solid-phase synthesis. The cationic AVPI-LMWP could condense pTRAIL by electrostatic interaction. The physical-chemical properties of the AVPI-LMWP/pTRAIL complexes were characterized. The cellular uptake efficiency and the inhibitory activity of the AVPI-LMWP/pTRAIL complexes on tumor cell were also performed. The results showed that the AVPI-LMWP/pTRAIL complexes were successfully prepared by co-incubation. With the increase of mass ratio (AVPI-LMWP/DNA), the particle size was decreased and the zeta potential had few change. Agarose gel electrophoresis showed that AVPI-LMWP could fully bind and condense pTRAIL at a mass ratio above 15:1. Cellular uptake efficiency was improved along with the increased ratio of W(AVPI-LMWP)/WpTRAIL. The in vitro cytotoxicity experiments demonstrated that the AVPI-LMWP/pTRAIL (W:W = 20:1) complexes was significantly more effective than the pTRAIL, AVPI-LMWP alone or LMWP/pTRAIL complexes on inhibition of HeLa cell growth. Our studies indicated that the AVPI-LMWP/pTRAIL co-delivery system could deliver plasmid into HeLa cell and induce tumor cell apoptosis efficiently, which showed its potential in cancer therapy using combination of apoptoic peptide and gene drugs.
Antineoplastic Agents ; chemistry ; Cell-Penetrating Peptides ; chemistry ; DNA ; chemistry ; Drug Delivery Systems ; HeLa Cells ; Humans ; Neoplasms ; drug therapy ; Particle Size ; Plasmids

AIM:To evaluate the efficacy and safety of intravitreous injection of Conbercept for macular edema secondary to macular branch retinal vein occlusion(MBRVO).METHODS:Clinical records of 19 patients(19 eyes) who were diagnosed MBRVO with macular edema (ME) in our hospital,from July 2015 to September 2016 were retrospectively analysed.All patients were treated with an intravitreal dose of 0.5mg or 0.05mL conbercept ophthalmic injection by the 3+ pro re nata (PRN) project.All patients were evaluated by best corrected visual acuity (BCVA),central retinal thickness(CRT),the number of the injection,and the complications.RESULTS:During the 1,2,3 and 6mo after treatment the mean BCVA all improved significantly(P＜0.01),and the mean CRT reduced statistical significantly(P＜0.01);3 eyes had refractory ME,and micro-aneurysm leakage were identified by FFA,and the ME was suppressed effectively after local laser photocoagulation.No complications,such as secondary vitreous haemorrhage,retinal detachment,persistent high intraocular pressure and endophthalmitis were observed during subsequent follow-up.CONCLUSION:Intravitreous injection of conbercept for macular edema secondary to MBRVO may reduce macular edema and improve visual acuity effectively and safely in the short term.

Objective To investigate the role and mechanism of matrix metalloproteinase 9(MMP-9) and tumor necrosis factor α (TNF-α) in the pathogenesis of allergic rhinitis (AR) and asthma (AS). Methods The rat models of AR and AS were made by injecting ovalbumin. The infiltration of eosinophils and mast cells were detected by hematoxylin-eosin (HE) staining and toluidine blue staining respectively, and the expression of MMP-9 and TNF-α in nasal mucosa and lung tissue were examined by immunohistochemical staining ( SP method). The relationship of their expression with upper and lower respiratory tract inflammation was analyzed. SPSS 13.0 software was used to analyze the data. Results The numbers of MMP-9 positive inflammatory cells in nasal mucosa and lung tissue of AR were ( 154.8±12.0)and (124. 0 ±8.2), (43. 2 ±7.6) and (34. 5 ±5.0) in the control groups, the difference was significant (t value were 24. 260, 29. 525 respectively, all P＜0.05). The numbers of MMP-9 positive inflammatory cells in nasal mucosa and lung tissue of AS were (149.9±11.7)and(120.1±7.3), (48.6 ± 7. 6) and (39.1±5.2)in control groups, the difference was significant (t value were 22. 929 and 28. 530respectively, all P＜0.05).The numbers of TNF-α positive inflammatory cells in nasal mucosa and lung tissue of AR were (188.8±17.0), and (134.8±7.9), (57.6±23.3)and(40. 3 ± 8. 2 ) in control groups, the difference was significant (t value were 13. 836 and 26. 220, all P ＜0.05). The numbers of TNF-α positive inflammatory cells in nasal mucosa and lung tissue of AS were (179. 2 ± 15.4 ) and ( 153. 5 ± 10. 1 ), (70. 5 ±33. 1 ) and ( 33.8 ± 14. 0) in control groups, the difference was significant ( t value were 9. 412 and 21. 858, all P ＜0. 05). There was a correlation between the expression of MMP-9 and TNF-α in nasal mucosa and lung tissue of AR ( r values were 0. 893 and 0. 700 respectively, P values were 0. 001 and 0. 024, respectively ) and AS ( r values were 0. 692 and 0. 644 respectively, P values were 0. 027and 0. 044 respectively) groups. Conclusions The inflammation is similar between AR and AS. The MMP9 and TNF-α may play an important role in the pathogenesis of upper and lower respiratory tract inflammation.

OBJECTIVEFor clarifying the situation of the natural infection of rodents having hemorrhagic fever with renal syndrome (HFRS) virus and to type Hantavirus (HV) using molecular technique in Shenzhen city in 2005, and offering guidance for prevention and control of HFRS.

METHODSData on the host animals was collected from the city of Shenzhen. ELISA and indirect immunofluorscent antibody(IFA) test were applied to the specific antibodies against HV in the sera of captured rats. Direct immunofluorscece assay was adopted to determine HFRS antigens and the lung tissues of the HV infected rats were inoculated into Meriones unguiculata to isolate HV. The whole viral RNA was extracted from the lung tissues of the HV infected rats and amplified the partial M fragments with RT-nested-PCR, using the HV genotype specific primers. The amplified genes were then sequenced, and subjected to genotyping and homology analysis.

RESULTS472 rodents were captured from Shenzhen in 2005. Surveillance on rats demonstrated 9.96% rats carrying HV (with a density of 8.25%) and the main host was Rattus norvegicus. In the blood samples of rats, anti-HV IgG antibodies were detectable in 56 cases by IFA, and proved to be positive in 76 cases by ELISA. We successfully isolated a HV strain designated as SZ2083 from Rattus norvegicus for the first time in Shenzhen and was identified to SEO type by RT-nested-PCR. Compared with the coding region of the M gene of HV L99 virus strain, the homologies of nucleotide among them were 97%, but the homology was 76% of the SZ2083 with HTN 76-118 virus strain.

CONCLUSIONResults showed the existence of natural epidemic areas of HFRS in Shenzhen city. Based on the results of sequencing, it is possible that the Seoul strain of HV might be the predominant serotype of virus harbored.

Animals ; China ; epidemiology ; Cities ; Data Collection ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique, Indirect ; Genotype ; Hantavirus ; classification ; genetics ; isolation & purification ; Hantavirus Infections ; epidemiology ; veterinary ; Rats ; virology ; Rodentia ; virology

Objective To explore the value of seeking tumor cells through cerebrospinal fluid in the metastasis and prognosis of retinoblastoma.Methods Two hundred and fifty-six cases clinic diagnosed retinoblastoma were collected,154 cases were boys and 102 cases were girls.There were 85 children with bilateral disease and 171 children with unilateral disease.Lumbar puncture was examined before treatment,compare the result and prognosis.Children who were found tumor cells in their cerebrospinal fluid were treated with 6 to 9 cycles of systemic chemotherapy with CTV(carboplatin,teniposide and vincristine)protocol and 8 to 10 intrathecal injections with cytrarabine,methotrexate and dexametha-sone.Follow-up were between 8 to 23 months,the mean time was 14.6 months.Results There were 8 cases found tumor cells in their cerebrospinal fluid in all 256 children,one of them died of wide spread intracranial metastasis;the other patient was given high dose chemotherapy with autologous perip-heral stem cell transplantation(APBSCT) because of intracranial metastasis,follow-up to now the patient′s condition was stable.The rest 6 children were enucleated in follow-up period,there were 4 cases(67%) with the histopathology of the eye indicates spread of tumour cells beyond the lamina cribrosa,and 2 cases with invading the cut end of the optic nerve.Six children were no metabasis evidences at other tissues.Conclusions Intracranial metastasis is the common metastasis style in retinoblastoma,so lumbar puncture to seeking tumor cells was important to evaluate intracranial metastasis,early treatment and prognosis.

Child ; Child, Preschool ; Combined Modality Therapy ; Female ; Germinoma ; blood ; therapy ; Humans ; Infant ; Male ; alpha-Fetoproteins ; analysis

OBJECTIVETo study the immunomodulatory effects and mechanisms of mesenchymal stem cells (MSC) derived from the bone marrow in acute leukemia patients in vitro.

METHODSBone marrow mononuclear cells from acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) were obtained and cultured in low serum medium. The immunophenotypes were assessed by FACS and immunol histochemistry. The levels of cytokines were evaluated by enzyme linked immunosorbant assay (ELISA). T-cell suppression ability was evaluated by Transwell chamber assay. Moreover, the immunoregulatory ability of AML- and ALL-derived MSC was detected by mixed lymphocyte culture assay.

RESULTSALL-derived MSC showed a typical fibroblast-like morphology. They were positive for CD29, CD44 and CD105, the positive rate were 98.81%, 99.25% and 90.52%, respectively, while negative for CD31, CD45 and CD34. Moreover, ALL- and AML-derived MSC didn't express HLA-DR and co-stirnulatory molecules (CD40, CD80 and CD86). ALL and AML derived MSC could secret several cytokines, such as TGF-β1 (567.58 ± 52.64 and 357.15 ± 33.52), HGF (647.27 ± 102.54 and 219.67 ± 62.37), IL-6 (59.67 ± 15.69 and 54.35 ± 12.31) and IL-11 (102.58 ± 23.54 and 78.21 ± 9.67), the level of secretion of TGF-β1 and HGF were higher in ALL bone marrow derived MSC than that of in AML bone marrow derived MSC. ALL and AML derived MSC significantly suppressed T lymphocyte proliferation in a dose-dependent manner, the counts per minute (CPM) were (3.58 ± 0.54) × 10(4), (2.87 ± 0.33) × 10(4), (1.78 ± 0.51) × 10(4) and (1.15 ± 0.15) × 10(4) for AML derived MSC, and CPM were (1.96 ± 0.31) × 10(4), (1.57 ± 0.28) × 10(4), (0.91 ± 0.41) × 10(4) and (0.22 ± 0.11) × 10(4) for ALL derived MSC when MSC were 0.5 × 10(4), 1 × 10(4), 2 × 10(4) and 5 × 10(4). In addition, the CPM was (4.01 ± 0.72) × 10(4) in control group. The immunosuppressive ability was different between MSCs derived from AML and ALL. The immunosuppressive effect of ALL derived MSC could be reversed by anti-TGF-β1 and anti-HGF antibody.

CONCLUSIONALL-derived MSC show immunoregulatory effect in vitro and this effect is achieved through cytokines. But MSCs derived from AML display abnormal changes in T-cell suppression ability.

Bone Marrow ; immunology ; Bone Marrow Cells ; cytology ; Cell Proliferation ; drug effects ; Cytokines ; pharmacology ; Humans ; Immunophenotyping ; Interleukin-11 ; metabolism ; Interleukin-6 ; metabolism ; Leukemia, Myeloid, Acute ; immunology ; Mesenchymal Stromal Cells ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo investigate the efficacy of high dose chemotherapy combined with autologous peripheral blood stem cell transplantation (APBSCT) for the treatment of neural ectodermal solid tumor originated from neural crest in children.

METHODSTwenty-three children at a medium age of 5.8 + or - 3.5 years with neural ectodermal solid tumor originated from neural crest were enrolled. Of the 23 children, 20 with stage IV neuroblastoma (9 were in complete remission, 7 were in partial remission and 4 were in progressive disease), 2 with stage IV primitive neuroectodermal tumor (PNET) in complete remission, and 1 with retinoblastoma in partial remission. Before APBSCT the children received 8.0 + or - 4.3 courses of chemotherapy. During chemotherapy the autologous peripheral blood stem cells were harvested and the tumor excision was performed. Then APBSCT was performed.

RESULTSThe reconstruction of the hematopoietic system was noted in 19 of 20 children with stage IV neuroblastoma 16.5 + or - 0.9 days after transplantation. A follow-up (median 15.8 months) was done in these children. The follow-up showed that the survival rate in children in complete remission before transplantation was 100%, 57% in those in partial remission, and none of children in progressive disease survived (P<0.05). The total survival rate was 67% in children with neuroblastoma. The child with retinoblastoma had complete remission in a 6-months follow-up. The tumors recurred in children with PNET 5 to 8 months after transplantation and all died within one year after transplantation.

CONCLUSIONSHigh dose chemotherapy combined with APBSCT can result in a good outcome in children with neural ectodermal solid tumor originated from neural crest in complete remission before transplantation and can improve the outcome in patients in partial remission before transplantation. However, the children with PNET, even in complete remission before transplantation, do not respond to the therapy.

Antigens, CD34 ; analysis ; Antineoplastic Agents ; therapeutic use ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Humans ; Male ; Neural Crest ; pathology ; Neuroblastoma ; therapy ; Neuroectodermal Tumors, Primitive ; therapy ; Peripheral Blood Stem Cell Transplantation ; Transplantation, Autologous

Adult ; Female ; Hepatitis B, Chronic ; complications ; physiopathology ; Humans ; Liver ; physiopathology ; Liver Function Tests ; Male ; Retrospective Studies ; Severe Acute Respiratory Syndrome ; complications ; physiopathology