Caused by Helminthosporium carposaprum, tomato brown lea f spot was a serious disease in green house in Henan Province. The condition for promoting sporulation of fungi were tested in this paper. The results showed th at the number of sporulation were different on the different medium,the fungi c ould sporulate a lot on the PDA+tomato leaf and Czapek medium, but V8、PSA and t omato juice restrained sporulation.The best carbon source and nitrogen source f or the fungi promoting sporulation were fructose and ammonium chloride respectiv ely,mannitol and Peptone ammonium sulfate restrained sporulation. Light and ult raviolet radiation were in favor of sporulation , ultraviolet radiation irradiat ing for 60～80min promoted sporulation. The fungi were promoted sporulation on the condition of lower or higher temperature and alkalescence,which 15℃o r 30℃,pH 8～9.

OBJECTIVETo explore the intervention of baicalin on signal transduction and activating transcription factor expression of ulcerative colitis (UC) patients.

METHODSRecruited were UC patients at Outpatient Department of Digestive Disease, Inpatient Department of Digestive Disease, Center for Digestive Endoscopy of College City Branch, Guangdong Provincial Hospital of Traditional Chinese Medicine, and Southern Hospital affiliated to Southern Medical University from June 2010 to January 2011. They were assigned to the UC group (33 cases) and the diarrhea-predominant irritable bowel syndrome (IBS-D) group (30 cases). Another 30 healthy subjects were recruited as a healthy control group. Peripheral blood mononuclear cells (PBMCs) in vitro intervened by different concentrations baicalin were taken from UC patients. IL23R gene expressions in vitro intervened by different concentrations baicalin were detected using Q-PCR. Expressions of signal transducer and activator of transcription 4 (STAT4) , STAT6, phosphorylated-STAT4 (p-STAT4), and p-STAT6 were detected using Western blot. Serum levels of IFN-γ, IL-4, IL-6, and IL-10 were measured by ELISA. Effects of different concentrations baicalin on expressions of PBMCs, and levels of IFN-γ, IL-4, IL-10 of UC patients were also detected.

RESULTSCompared with the negative control group, 40 µmol baicalin obviously decreased IL23R gene expression of UC patients (P <0. 01). Compared with the healthy control group and the IBS-D group, p-STAT4/STAT4 ratios increased, p-STAT6/STAT6 ratios decreased, levels of IFN-γ, IL-4, IL-10 all increased in the US group (all P <0. 05). Compared with the negative control, 5 and 10 µmol baicalin groups, 20 and 40 moL baicalin obviously decreased p-STAT4/STAT4 ratios (all P <0. 05); 20 and 40 µmoL baicalin obviously increased p-STAT6/STAT6 ratios (all P <0. 05); 20 and 40 µmoL baicalin obviously lowered levels of IFN-γ and IL-4, and elevated IL-10 levels (all P <0. 05).

CONCLUSION40 µmoL baicalin could in vitro inhibit p-STAT4/STAT4 ratios, adjust p-STAT6/STAT6 ratios and related cytokines, thereby balancing the immunity and relieving inflammatory reactions of UC.

Activating Transcription Factors ; metabolism ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Blotting, Western ; Colitis, Ulcerative ; drug therapy ; metabolism ; Cytokines ; metabolism ; Flavonoids ; therapeutic use ; Humans ; Interleukin-10 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Irritable Bowel Syndrome ; drug therapy ; metabolism ; Leukocytes, Mononuclear ; Medicine, Chinese Traditional ; Phosphorylation ; STAT6 Transcription Factor ; metabolism ; Signal Transduction

A fragment containing amino acid residues 561～578 of HSV-2 glycoprotein G(gG2) was obtained by PCR assembling technique,and doubly cloned into vector pET-KDO.The recombinant plasmid was transformed to BL21(DE3)plysS.Fusion protein,of molecular weight about 39kDa was highly expressed by induction of IPTG.Western blot result showed the fusion protein had good antigenicity.After putification and digestion,the purity reached 95%.The digested purified protein was analysed by ELISA and showed good sensitivity and specificity.The recombinant protein should be useful for type-specific serodiagnosis of HSV-2.

To explore methods of maintaining the self-renewal capacity of hematopoietic stem cells, inhibiting their overdue differentiation and expanding hematopoietic cells massively, the murine bone marrow stromal cells were coated on microcarriers, then co-cultured with hematopoietic cells from murine bone marrow as group 2 (G2). The G2 contents were wrapped by sodium alginate, then cultivated as group 1 (G1). The only microcarriers coated with stromal cells as group 3 (G3) and the only bone marrow cells as group 4 (G4) were cultivated as control groups. Contrasting observation and microphotograph were performed; the number of total marrow cells, the colony efficiency of CFU-GM and the percentages of CD34(+) cells were determined. Three repeated experiments indicated that the colony efficiency of CFU-GM before culture (G0) were 118.8 +/- 38.1/10(5) marrow cells, and the total outputs of CFU-GM (G0) were 9 501.3 +/- 3 049.0. After culture for two weeks, hematopoietic cells were adhered to or embedded in stromal cells coating the microcarriers, and had formed hematopoietic focus. The colony efficiency of CFU-GM per 10(5) mononuclear cells in group G1, G2, G3 and G4 averaged 30.9 +/- 13.7, 147.3 +/- 66.0, 23.4 +/- 23.1 and 15.9 +/- 8.1, respectively; the total outputs of CFU-GM in group G1, G2, G3 and G4 averaged 273.8 +/- 75.3, 9 424.8 +/- 7 933.7, 419.1 +/- 305.6 and 140.7 +/- 20.7, respectively; the measured CFU-GM output in group G2 was significantly higher than that in group G4, and still significantly higher than the sum of groups G3 and G4 (t = 6.553, t = 5.494; P < 0.05). The percentage of CD34 cells before culture was 10.0 +/- 1.0; after cultuer for two weeks, the percentages of CD34(+) cells in G1, G2, G3 and G4 averaged 4.0 +/- 1.0, 11.0 +/- 1.0, 3.3 +/- 1.5 and 2.2 +/- 0.8, respectively. The percentage of CD34 positive control (3T3 cells) was 17.0 +/- 1.0. This result was consistent with the result of CFU-GM outputs measured. These data suggest that microcarriers coated with stromal cells can perfectly support the ex vivo hematopoiesis at least to four weeks, while hematopoietic cells fixed by alginate are not significantly different from control groups. The hematopoiesis-simulating model of microcarriers is successful, whereas the hematopoiesis-simulating model of alginate macrocarriers can not support the ex vivo hematopoiesis.

OBJECTIVETo investigate the feasibility of prevention and treatment of early scar through observing the effect of feeding immunosuppressive drug cyclophosphamide on rabbit ears hypertrophic scar tissue in early stage.

METHODSThirty-two Rabbit ears were used to establish animal models for hypertrophic scar and randomly divided into four groups: group of distilled water (A), group of cyclophosphamide 5 mg x kg(-1) x d(-1) (B), group of 10 mg x kg(-1) x d(-1) (C), group of 30 mg x kg(-1) x d(-1) (D). Before animal models were built and after administration for 14 days, 28 days, leukocytes and lymphocytes were detected. After 28 days, specimens were harvested and underwent HE staining and VG staining in order to assess HI, NA, AA value changes. The data (HI, NA, AA) from each group were compared by analysis of variance, and the variance for the rank sum test when missing.

RESULTSOn the 14th day, the number of leukocytes in group A, B, C, D were (8.62 +/- 0.58) x 10(9)/L, (4.48 +/- 0.41) x 10(9)/L, (2.7 +/- 0.26) x 10(9)/L, (1.33 +/- 0.27) x 10(9)/L; the number of lymphocytes in group A, B, C, D were (3.11 +/- 0.21) x 10(9)/L, (1.67 +/- 0.16) x 10(9)/L, (0.42 +/- 0.10) x 10(9)/L, (0.40 +/- 0.09) x 10(9)/L. On the 28th day, the number of leukocytes in group A, B, C, D was (8.63 +/- 0.53) x 10(9)/L, (5.10 +/- 0.27) x 10(9)/L, (3.10 +/- 0.26) x 10(9)/L, (1.98 +/- 0.20) x 10(9)/L; the number of lymphocytes A, B, C, D was (3.06 +/- 0.16) x 10(9)/L, (2.08 +/- 0.14) x 10(9)/L, (0.96 +/- 0.19) x 10(9)/L, (0.14 +/- 0.07) x 10(9)/L. On the 14th day and 28th day, the number of leukocytes and lymphocytes in experimental groups was reduced, showing a negative relation with cyclophosphamide dose (P < 0.05). The HI in group of A, B, C, D was 3.02 +/- 0.24, 2.59 +/- 0.43, 2.06 +/- 0.19, 1.63 +/- 0.11; the AA was 40.49 +/- 2. 07, 35.29 +/- 1.99, 28.36 +/- 1.87, 24.99 +/- 1.82; the NA was 4570.5 +/- 259.3, 4222.5 +/- 199.6, 3540.3 +/- 170.3, 3341.4 +/- 228.8. The difference in HI, AA, NA between control group and any of the experimental groups was statistically significant (P < 0.01). Each group, with the dose increased, except NA content of group C and D, the HI, AA, NA was more smaller, negative correlation, the difference was statistically significant (P < 0.05).

CONCLUSIONSFeeding cyclophosphamide can inhibit leukocytes and lymphocytes number, so as to inhibit the proliferative activity of hypertrophic scar. It has significant effect on prevention of hypertrophic scar on rabbit ears in early stage.

Animals ; Cicatrix, Hypertrophic ; drug therapy ; prevention & control ; Cyclophosphamide ; pharmacology ; Ear ; pathology ; Female ; Leukocyte Count ; Leukocytes ; drug effects ; Lymphocyte Count ; Lymphocytes ; drug effects ; Male ; Rabbits

OBJECTIVETo observe the effect of hyperbaric oxygen on the scar formation at the rabbit ears at an early stage.

METHODS16 New Zealand rabbits were used to establish the hypertrophic scar model on the ears, 4 wounds on each ear. The rabbits were randomly divided into hyperbaric group(n = 8) and control group (n = 8). The rabbits in the hyperbaric group received hyperbaric oxygen treatment, inhaling oxygen for 1 hour daily under 2ATA circumstance until the wounds were healed. The wound healing and the scar size, thickness, color and hardness in the ears were recorded. After healing, the scar was taken for histologic study with HE staining, Masson staining and trinitrophenol sirius red staining.

RESULTSIn hyperbaric oxygen group, the healing time of 64 wounds was (16.7 +/- 1.8) d, while it was (20.2 +/- 2.3) d in the control group, suggesting a significant difference between two groups (P < 0.05). The incidence of hyperplastic scar was lower (38/64, 59. 4% ) in hyperbaric oxygen group than that (52/ 64, 81.2%) in control group with significant difference between them (P < 0.05). Compared with the control group, the hyperbaric oxygen group had thicker corium layer and less fibroblasts, well-arranged but rare collagen and less collagen nodus and vortex-like structure under microscope. The hyperplastic index of scar was 3.48 +/- 0.94 in hyperbaric oxygen group and 4.65 +/- 0.76 in control group respectively (P < 0.01). The density of fibroblast in two groups were 186.5 +/- 27.3 (hyperbaric oxygen group) and 246 +/- 41.6 (control group) with statistically significant difference (P < 0.05). Furthermore, the area density of collagen fiber were (31.42 +/- 5.36)% in hyperbaric group and (43.62 +/- 7.36)% in control group (P < 0.05). The amount of collagen I and III was (71.42 +/- 5.36)% and (28.58 +/- 5.36)% in hyperbaric oxygen group, (62.46 +/- 7.32)% and (37.54 +/- 7.32)% in control group (P < 0.05). The ratio of collagen I to III was 2.499 in hyperbaric oxygen group, which was similar to the ratio (4:1) in normal skin, compared with the control group (1.664).

CONCLUSIONSThe hyperbaric oxygen can promote wound healing and effectively inhibit the early hyperplastic scar in rabbits ears.

Animals ; Cicatrix ; Disease Models, Animal ; Ear ; pathology ; Female ; Hyperbaric Oxygenation ; Male ; Rabbits ; Wound Healing

Objective To explore the characteristics of human plague using a sero-epidemiologic method in the source of the three rivers area in Qinghai for possible plague control strategies. Methods Investigate human plague sero-epidemiologically in the source of 4 counties in the three rivers area in Qinghai. The human serum would be tested to confirm the sew-positive rate for plague F1 antibody using indirect hemagglutination assay(IHA). Results A total of 2508 local participants were tested in 4 counties, the overall plague sero-positive rate was 2.31%(58/2508). This represents a statistically significant difference with 4 counties(X2=19.30,P<0.01). The sew-positive rate for males and females were 2.54% (32/1261) and 2.09% (26/1247), respectively. There were no statistically significant differences between males and females(X2= 0.65,P 0.05). The sero-positive rate in herdsman, cadre, Tibetan, Hart nationalities were 3.54% (44/1243), 6.47% (11 / 170), 2.40% (56/2335) and 1.47% (2/136), respectively. The sero- positive rate increased with age. The highest titre for human plague serum antibody was 1 : 640. Conclusion There were occult infections of plague in the population on source of three rivers area in Qinghai. Sero-epidemiologic data revealed that the human plague sero-positive rate was closely correlated with the local animal plague.

OBJECTIVETo investigate the effect of the gene nanoparticles using chitosan (CNP), arginine modified chitosan (ANP), or hexadecylated chitosan (HNP) as carriers on the human normal liver cell line L02.

METHODSCNPs, ANPs, and HNPs were prepared using complex coacervation method. The size and zeta potential of the gene nanoparticles were measured using Zetasizer nanoZS. The nanoparticles at concentrations of 5, 10, 30, and 50 microg/ml (based on the content of DNA) were incubated with L02 cells, respectively. The cell viability was evaluated by MTT assay, and the effect of the gene nanoparticles on the cell apoptosis was analyzed by flow cytometry.

RESULTSThe zeta potential of the gene nanoparticles ranged from 12.10 to 14.63 mV, and their diameters ranged from 148.07 to 179.47 nm. MTT assay showed that the viability of L02 cells began to decrease when the concentration of CNPs reached 30 microg/ml and higher. Furthermore, the CNPs could induce cell apoptosis as the concentration of CNPs reached 30 microg/ml and higher.

CONCLUSIONCNPs can induce L02 cell apoptosis at relatively higher concentrations.

Apoptosis ; Cell Line ; Cell Survival ; Chitosan ; chemistry ; DNA ; chemistry ; genetics ; Gene Transfer Techniques ; instrumentation ; Humans ; Nanoparticles ; chemistry

OBJECTIVEThe presence of liver fibrosis in patients with beta-thalassemia major has been demonstrated to be an important negative prognostic factor. Identification of liver fibrosis in early stage would be of great value. Hyaluronic acid (HA), type III pre-collagen (PC III), collagen IV (C IV) and laminin (LN) as serum markers were widely used in the diagnosis of liver fibrosis in patients with chronic viral infections or alcoholic liver diseases. However, their values in thalassemic liver fibrosis have not been studied. This work was to determine the serum HA, PC III, C IV and LN levels in children with beta-thalassemia major and evaluate the diagnostic utility.

METHODSerum HA, PC III, C IV and LN in 49 hospitalized children with beta-thalassemia major (aged 1 - 15 years with the media age of 6.27 years) and 41 healthy children served as controls (aged 1 - 13 years with media age of 6.40 years) were detected by radioimmunoassay (RIA). Forty-five of 49 cases were performed percutaneous liver biopsies, and the histopathological fibrosis was compared with the four serum markers. The correlation and discriminate analysis were used.

RESULTSAll the serum levels of HA, PC III, C IV and LN in beta-thalassemia were significantly higher than those in controls (P < 0.01). In 36 of 45 cases, the histopathology showed liver fibrosis including stage I and stage II by biopsies with a positive rate of 80%. The serum levels of four markers increased successively with the aggravation of liver fibrosis from stage 0 to stage II, and significant correlation was observed between the level of HA or PC III and the stage of fibrosis (HA, r = 0.379, P = 0.017; PC III, r = 0.455, P = 0.04). While there was no difference between the level of C IV or LN and fibrosis (C IV, r = 0.312, P = 0.053; LN, r = 0.310, P = 0.055). Using discriminate analysis, the discriminate function of co-detection of the four markers for the diagnosis of fibrosis was 0.002 HA + 0.003 PC III + 0.002 C IV + 0.006 LN-1.859, which had a sensitivity of 93.88%, specificity of 68.29%, predictive value of positive test and negative test of 77.97% and 90.32%, respectively. Moreover, there was a significant correlation between the serum level of HA or PC III and the liver iron concentration (HA, r = 0.318, P = 0.035; PC III, r = 0.305, P = 0.044).

CONCLUSIONThe results suggest that, in beta-thalassemia major with chronic liver disease, HA and PC III showed more practical value in diagnosing liver fibrosis than the levels of C IV and LN. The combination of the four serum markers could improve the accuracy and reliability of the diagnosis. A validation study is necessary before introducing into the prediction function during the clinical practice.

Adolescent ; Biomarkers ; blood ; Child ; Child, Preschool ; Collagen Type III ; blood ; Collagen Type IV ; blood ; Female ; Humans ; Hyaluronic Acid ; blood ; Infant ; Laminin ; blood ; Liver Cirrhosis ; blood ; complications ; diagnosis ; Male ; Prognosis ; beta-Thalassemia ; blood ; complications ; pathology