Background Scarring of surgical area,the most important factor,leads to the failure of glaucoma filtering surgeries.Therefore,more and more attentions are paid to the causes and process of scar formation.Objective This study was to compare the differences of proliferation and migrating abilities of fibroblasts between filtering bleb scar tissue and normal Tenon capsular tissue,and to investigate the inhibitory effects of miRNA-200a (miR-200a) on biological behavior of conjunctival fibroblasts.Methods Normal Tenon capsular tissue and filtering bleb scar tissue were collected during the strabismus surgery and glaucoma filtering surgery,respectively for the primarily culture of fibroblasts.The proliferation (absorbency,A) of the cells was assayed by cell counting kit-8 (CCK8) method;the relative migrating distance of the cells was measured by cell scratch test;and the relative expressions of transforming growth factor-β1 (TGF-β1)mRNA and miR-200a mRNA in the cells were detected by realtime fluorescence quantitative PCR.TGF-β1 mimic of 0,1,2 and 5 ng/ml was added in the medium of human normal Tenon capsular-derived fibroblasts (HTFs),and 0.00,0.25,0.50,1.00 μg/ml TGF-β1 inhibitor was added in the medium of human scarring-derived fibroblasts (HSFs) for 24 hours,respectively,and CCK8 was used to evaluate the proliferation of the cells.The relative migrating distance as well as the relative expressions of miR-200a mRNA were analyzed in the 2 ng/ml TGF-β1 mimic-or 1.00 μg/ml TGF-β1 inhibitor-treated cells.Results The primary conjunctival presented the spindle and star-like in shape with large body and oval nuclei.The cells showed the positive response for keratin and vimentin antibodies.The A values were 1.476±0.110 in the HSFs and 0.958±0.074 in the HTFs,with a significant difference between them (t =24.900,P=0.016).The relative expressions of TGF-β1 mRNA were significantly higher in the HSFs than those in the HTFs,and the relative expressions of miR-200a were evidently lower in the HSFs than those in the HTFs,showing significant differences between them (t =6.358,P =0.024;t=7.394,P =0.018).Compared with the 2 ng/ml TGF-β1 mimic-treated HTFs,the relative migrating distance increased,while the expression level of miR-200a mRNA was significantly reduced in the 2 ng/ml TGF-β1 mimictreated HSFs (all at P＜0.05);Compared with the 1.00 μg/ml TGF-β1 inhibitor-treated HTFs,the relative migrating distance decreased,but the expression level of miR-200a mRNA was significantly elevated in the 1.00 μg/ml TGF-β1 inhibitor-treated HSFs (all at P＜0.05).Conclusions The proliferation and migrating abilities are stronger in the HSFs than those in the HTFs,which probably is regulated by the expression of miR-200a in the cells.The miR-200a plays a negative feedback for the effect of TGF-β1 promoting proliferation and migration of fibroblasts.

Background5-Nitro-2-(3-styrene-acrylic amine) benzoic acid ( NPPB),a chloride channel inhibitor,has a promoting effect on cell apoptosis in myocardial ischemia and reperfusion of domestic rabbit.The CIC chloride channel has been found in the ocular trabecular cells.However,the effect of NPPB on the shape and function of trabecular cells is unclear. Objective This study was performed to investigate the effect of NPPB on the proliferation,cell cycle progression and apoptosis of human trabecular meshwork cells.MethodsThe immortalized human trabcular cell strain was cultured,and logarithmic-phase cells were incubated in 96-well plates at a density of 1 ×106/ml.Different concentrations of NPPB (10,50,100 μ mol/L) were added to the medium,and the MTT assay was used to assess the growth and proliferation of the cells.Flow cytometry was used to evaluate the cell cycle.Then,100 mg/L 5-FU or 100 mg/L 5-FU + 100 μmol/L NPPB was used to induce cell apoptosis,which was assessed by Annexin V-PI.The membrane potential of mitochondria was examined using rhodamine 123 (△ψm).Results After 48 hours of treatment with NPPB,the abosorbency (A value) of the cells was gradually lowered with the increasing dose of NPPB,with significant differences among the 4 groups (F =7.230,P =0.006).Compared with the 10 μmol/L NPPB group,the A values were significantly declined in the 50 and 100 μmol/L NPPB groups (t =1.610,P =0.025 ;t =12.270,P =0.001 ).Forty-eight hours after exposure to NPPB,the percentage of cells in G0/G1 phase was increased and that in the S phase was decreased.The percentages of cells in different phases of cell cycle were significantly different in comparison with their control groups (without NPPB)( P＜0.05 ).Twenty-four and 48 hours after the treatment with 100 mg/L 5-FU,the apoptosis rates of the cells were raised in the 100 mg/L 5-FU group and 100 mg/L 5-FU + 100 μmol/L NPPB group compared to the without NPPB group (t24h =2.130,P =0.023;t48h =4.810,P=0.011 ) ;while that in the 100 mg/L 5-FU+100 μmol/L NPPB group was higher than the 100 mg/L 5-FU group ( t24 h =1.980,P =0.037 ; t48 h =1.290,P =0.028 ),and the mitochondrial membrane potential was lowered ( t24h =1.580,P =0.029 ; F48 h =6.200,P =0.015 ).Conclusions NPPB suppresses the proliferation of human trabecular cells and promotes the cells to enter S phase via the G1/S check point.In addition,ClC might be involved in an anti-apoptosis mechanism through the internal mitochondrial pathway.

Objective To explore the clinical value and typing characteristic of prenatal ultrasonography for fetal cystic adenomatoid malformation of the lung(CAML).Methods Ultrasonographic features and typing of fetal CAML in 41 cases detected by prenatal ultrasonography were analyzed retrospectively.All cases were followed up until to the induction of labor or birth.Results (1)Site of tumor: in the 41 cases,there were 22 cases on the left side,15 on the right side and 4 on bilateral.(2)Typing diagnosis of prenatal ultrasonography: three cases were CAML type Ⅰ,14 were type Ⅱ,and 24 cases were type Ⅲ.(3)Pathology diagnosis: thirty two cases were induced abortion.CAML was confirmed in 29 cases by autopsy and the classifications were consistent with the prenatal diagnosis of ultrasonography.Three pulmonary sequestration cases were prenatally misdiagnosed as CAML type Ⅲ by ultrasonography.(4)The other nine cases were followed up to birth.Three masses decreased gradually and then disappeared.Six newborns were confirmed as CAML by CT.Their typings were consistent with the postnatal diagnosis.(5)The diagnostic accuracy rate of prenatal ultrasonography for CAML was 92.7%(38/41).Its misdiagnostic rate was 7.3%(3/41).Conclusions Prenatal ultrasonography has a high accuracy rate for the diagnosis and classification of CAML and is the first choice to detect CAML early in pregnancy.It has an important clinical value.Pulmonary sequestration should be distinguished from CAML type Ⅲ because they tend to be confused.

A total of 152 strains of endophytic fungi were isolated from the barks of Eucommia ulmoides in three regions (Lueyang country, Zunyi country, Cili country). Based on morphological characteristics and analysis of ITS sequences, these strains were identified into 8 genera. Thereinto Phomopsis, Diaporthe and Alternaria were common genera to Eucommia barks from different sites. But the dominant genus was different: Alternaria was the dominant genus in the barks from Cili country, and Phomopsis was the dominant genus from Zunyi country, then Diaporthe was the one from Lueyang country. According to the similarity coefficient, the composition of the endophytic fungi was distinctly different between the barks from three sites. The diversity and species richness in Lueyang country and Cili country were found higher than those in Zunyi country. The evenness of endophytic fungi was 0.936 5 in Lueyang county, which was higher than 0.737 1 or 0.641 0 in Cili county or Zunyi county, respectively. After phylogenic analysis and calculating the genetic distances of typical strains belong to Phomopsis and its perfect stage--Diaporthe, there was very high genetic diversity in the two genera from our study. In conclusion, the community structure and diversity of endophytic fungi were significant different in Eucommia barks from the three habitats.
DNA, Fungal ; genetics ; DNA, Intergenic ; genetics ; Ecosystem ; Endophytes ; classification ; physiology ; Eucommiaceae ; microbiology ; Fungi ; classification ; genetics ; physiology ; Phylogeny ; Plant Bark ; microbiology

ObjectiveTo observe protective effects of agmatine (AGM) on inflammatory response and spleen immune function in mice with trauma.Methods Forty-eight adult male C57BL/6 mice were randomly divided into three groups (n= 16 each), including control group, model group (bilateral femoral fracture and removal of 35% of the total blood volume), and AGM group (trauma/hemorrhage & AGM 200 mg/kg). Eight mice in each group were sacrificed at 3 hours and 24 hours, respectively, after modeling, and blood samples and tissue homogenate of spleen and liver were collected. The contents of tumor necrosis factor-α (TNF-α), interleukins (IL-6, IL-1β) in serum and liver tissue were determined with enzyme linked immunosorbent assay (ELISA). Serum aspartate transaminase (AST), alanine aminotransferase (ALT) and lactic dehydrogenase (LDH) were determined with automatic biochemistry analyzer. Spleen proliferation response stimulated with concanavalin A (ConA) was evaluated with methyl thiazolyl tetrazolium colourimetry (MTT).γ-interferon (IFN-γ) and IL-2 releases were determined with ELISA.Results Compared with control group, 3 hours after trauma/hemorrhage, the levels of serum TNF-α, IL-6, and IL-1β in model group were significantly elevated [TNF-α (ng/L): 145.38±31.50 vs. 23.06±11.14, IL-6 (ng/L): 496.94±50.76 vs. 47.13±17.47, IL-1β (ng/L): 321.31±43.02 vs. 29.25±16.24,allP< 0.01]. It was found that AGM treatment could alleviate the increase in serum pro-inflammatory mediators induced by trauma/hemorrhage, such as TNF-α (ng/L:111.56±25.47 vs. 145.38±31.50), IL-6 (ng/L: 412.56±44.33 vs. 496.94±50.76), IL-1β (ng/L: 273.38±45.25 vs. 321.31±43.02,P< 0.05 orP< 0.01). Twenty-four hours after trauma/hemorrhage, serum pro-inflammatory mediators were recovered to the levels in control group. There was no significant difference in TNF-α and IL-6 levels at 3 hours after trauma/hemorrhage among groups. Compared with control group, the expressions of liver TNF-α and IL-6 in model group were increased at 24 hours following trauma [TNF-α (ng/mg): 32.93±4.90 vs. 26.58±2.33, IL-6 (ng/mg): 11.20±1.66 vs. 8.38±0.89,bothP< 0.01]. However, AGM inhibited the level of TNF-α (ng/mg:28.92±3.16 vs. 32.93±4.90) and IL-6 (ng/mg: 9.03±1.28 vs. 11.20±1.66) in the liver as induced by trauma/hemorrhage (P< 0.05 andP< 0.01). At 24 hours after modeling, model group and AGM group had distinctly higher serum AST, ALT, LDH levels than those of control group [AST (U/L): 405.9±31.2, 245.7±22.1 vs. 128.2±15.9; ALT (U/L): 92.1±6.3, 51.6±5.0 vs. 30.1±3.2; LDH (U/L): 606.7±36.3, 478.7±25.3 vs. 384.0±16.6, allP< 0.01]. Nevertheless,the increase in serum AST, ALT and LDH was alleviated in AGM group (allP< 0.01). Meantime, trauma/hemorrhage produced a noticeable depression of proliferation of splenic cells and IFN-γ and IL-2 release stimulated with ConA compared with control group [proliferation rate: (40.97±4.13)% vs. (89.99±7.76)%, IFN-γ(ng/L): 91.6±12.3 vs. 353.2±21.5,IL-2 (ng/L): 53.4±6.4 vs. 91.0±12.2,allP< 0.01]. In contrast, AGM notably restored the capacity of proliferation response of splenic cells [proliferation rate: (74.86±5.75)% vs. (40.97±4.13)%, P< 0.01],enhanced the release of IFN-γ and IL-2 stimulated with ConA [IFN-γ (ng/L): 327.8±23.6 vs. 91.6±12.3, IL-2 (ng/L): 74.8±10.4 vs. 53.4±6.4, bothP< 0.01].Conclusion AGM can dramatically alleviate spleen immunosuppression, excessive inflammation and organ damage induced by trauma/hemorrhage.

A mixed bacteria culture F6 isolated from oil field wastewater can degrade petroleum hydrocarbons efficiently. The bacteria were suitable to treat oil-polluted wastewater of oil field. Simulated result treating oil-polluted wastewater in laboratory showed that after "XingyiLian" wastewater of Liaohe Oil Field was treated by fluidized-bed bioreactor system with the vehicle of activated carbon , the amount of oil and CODcr of the flow out water were decreased from 45mg/L to 4. 1mg/L and 470mg/L to 42mg/L separately , according with first class standards of Chinese Wastewater Discharge Regulation.

Mice and 3-day-old chickens were orally inoculated with the recombinant attenuated Salmonella typhimurium strain ZJ111 carrying pcDNA3-F expression plasmid encoding the fusion protein of Newcastle disease virus (NDV). The results showed that ZJ111/pcDNA3-F was relatively safe. The recombinant plasmid pcDNA3-F was stable within the host stain ZJ111 in vitro and in vivo as shown by restriction enzyme analysis and PCR identification of the F gene. In an experimental vaccination study, 3-day-old chickens were orally immunized with ZJ111/pcDNA3-F with a dose of 108 cfu per chicken and boosted two weeks later. At week 4 post boosting, all chickens were challenged with a lethal dose of a virulent NDV strain F48 E9. The results showed that oral vaccination with ZJ111/pcDNA3-F induced stronger humoral and cellular immune responses than intramuscular immunization with naked pcDNA3-F plasmid. It also exhibited higher protection rate than the latter (66.7% vs 50%). This study indicates that the DNA vaccine using attenuated Salmonella typhimurium as delivery carrier had good safety, stability and immunogenicity and exhibited good potential of low cost and convenience for poultry disease control.
Animals ; Chickens ; Immunity, Cellular ; immunology ; Immunity, Humoral ; immunology ; Mice ; Newcastle Disease ; immunology ; virology ; Plasmids ; Polymerase Chain Reaction ; Salmonella typhimurium ; genetics ; metabolism ; Vaccines, DNA ; adverse effects ; genetics ; immunology

AIMTo prepare of isopropyl myristate (IPM) molecular gels and investigate of its transdermal capability.

METHODSMicrostructure of IPM gels was studied by scanning electron microscope (SEM) and optical microscope (OM). The rheology and thixotropy of IPM gels were investigated by viscosity. Triptolide was used as model drug to investigate its transdermal capability.

RESULTSThe microstructure of IPM gels was a three-dimension network formed by the aggregation of Span 60 in IPM, which was rod-like tubular aggregate. It has good rheology and thixotropy. There was a good linear correlation between the accumulative permeated amount per unit area and the time for triptolide-loaded IPM gels. The permeation process agreed with zero order pharmacokinetics. The average permeability through rat skin for triptolide was 19.26 ng x cm(-2) x h(-1), which was 2.92 times of triptolide unguents obtained commercially available.

CONCLUSIONIsopropyl myristate molercular gel can be formed by span 60 assemblies. Transdermal capability drug-loaded IPM gels was better than that of triptolide unguents.

Administration, Cutaneous ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; isolation & purification ; pharmacokinetics ; Diterpenes ; administration & dosage ; isolation & purification ; pharmacokinetics ; Drug Carriers ; Epoxy Compounds ; Male ; Mice ; Microscopy, Electron ; Myristates ; chemistry ; pharmacology ; Phenanthrenes ; administration & dosage ; isolation & purification ; pharmacokinetics ; Plants, Medicinal ; chemistry ; Rheology ; Skin Absorption ; drug effects ; Tripterygium ; chemistry ; Viscosity

OBJECTIVETo investigate the effects of Tpo and/or IL-11 gene modified stromal cells on the expansion of CD(34)(+) hematopoietic stem/progenitor cells in cord blood.

METHODSRetroviral vectors containing Tpo or IL-11 gene were constructed and used to transfect the stromal cell line HFCL. Tpo and/or IL-11 mRNA was assayed by Northern blot. Non-modified stromal cells were used, CD(34)(+) hematopoietic stem/progenitor cells from cord blood were expanded on gene-modified stromal cells for 7 days. The phenotype of CD(34)(+)CD(38)(-) primitive progenitors was detected by flow cytometry.

RESULTSHFCL expressed Tpo and/or IL-11 mRNA after transfected by the retroviral vectors. The percentages of CD(34)(+)CD(38)(-) primitive progenitors in the cultures of Tpo, IL-11 and Tpo + IL-11 modified HFCL were (1.8 +/- 0.24)%, (1.62 +/- 0.23)%, and (2.45 +/- 0.28)%, respectively, which were higher than that in the control [(0.8 +/- 0.23)%].

CONCLUSIONThe stromal cells modified by Tpo and/or IL-11 gene were able to enhance ex vivo expansion of CD(34)(+) and CD(34)(+)CD(38)(-) hematopoietic stem/progenitor cells from cord blood.

ADP-ribosyl Cyclase ; analysis ; ADP-ribosyl Cyclase 1 ; Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; physiology ; Humans ; Infant, Newborn ; Interleukin-11 ; genetics ; Membrane Glycoproteins ; Stromal Cells ; physiology ; Thrombopoietin ; genetics

Animals ; Female ; Interferon-alpha ; therapeutic use ; Interferon-gamma ; therapeutic use ; Liver Cirrhosis ; drug therapy ; parasitology ; Mice ; Mice, Inbred ICR ; Schistosoma japonicum ; Schistosomiasis japonica ; complications