Coloboma ; Humans ; Infant, Newborn ; Male ; Optic Nerve ; abnormalities

Objective To evaluate the effect of gamma irradiation on the proliferation,differentiation,and mineralization of murine osteoblastic cells,and to investigate the related molecular mechanism.Methods Osteoblastic cells were irradiated by different doses (0,0.5,1.0,2.0,5.0 Gy)of 137Cs γ-rays.Cell morphology was observed with a microscopy,cell viability was analyzed by MTT assay,and ALP activity was analyzed by the methods of enzyme histochemistry and PNPP.Meanwhile,gene expressions of ALP,osteocalcin (OC),collagen Ⅰ,osteoprotegerin (OPG) and receptor activator of nuclear factor-kB ligand (RANKL) were measured by semi-quantified RT-PCR.Results Cell viability decreased with the radiation doses over 1.0 Gy ( t =6.197 - 18.677,P ＜ 0.05 ).After radiation with a dose over 2.0 Gy,the cell number and the junctions of cell protrusions decreased,the cells had low refractivity and the activity and mineralization ability of ALP were also inhibited ( t =2.790 -2l.374,P ＜0.05).In addition,the expressions of ALP and OC mRNA were down-regulated significantly (t =3.563 -16.508,P ＜ 0.05) when the radiation dose was higher than 0.5 Gy,and the expressions of OPG,OPG/RANKL mRNA were down-regulated ( t =12.942,4.954,P ＜ 0.05 ) at 5 Gy.But the expressions of collagen Ⅰ and RANKL mRNA were not affected by irradiation.Conclusions The osteoblastic cells were significantly influenced by γ-irradiation,including morphological changes,inhibition of cell proliferation,differentiation and mineralization ability. Meanwhile,mRNA expressions of ALP and OC were downregulated.OPG/RANKL may be a main pathway of osteoblastic cell damage under high dose radiation.

To standardize the harvest ways of Dendrobium officinale and improve the quality and yield of D. officinale, a field experiment was carried out to study the effect of two kinds of harvest ways, which were keeping some of the axial shoot and harvesting all of the shoot by the end of the year. Then, the agronomic traits and yield were measured and the contents of polysaccharides and extractum were determined. The results showed that the harvest ways significantly affected the growth of D. officinale. Keeping some of the axial shoot could significantly improved the number of sprout, stem length, internode number and the internodal length, which also triggered increase the weight of fresh stems, leaves and the total of them and dry stems in per unit area, but it could not promote the stem diameter and the polysaccharide content in stems. Keeping some of the axial shoot moderately was conducive to the improvement of the production of medicinal materials in the process of harvesting by promoting the germination and growth of new buds, and to ensure the polysaccharide content by regulating the illumination and the density of cultivation.
Agriculture ; methods ; Dendrobium ; chemistry ; growth & development ; Drugs, Chinese Herbal ; analysis ; Plant Leaves ; chemistry ; growth & development ; Plant Stems ; chemistry ; growth & development

Objective To investigate the efficacy of precise hemihepatectomy guided by middle hepatic vein(MHV),and to study the value of preoperative hepatic vein evaluation.Methods The clinical data of 68 patients who received hemihepatectomy at the Nanjing Drum Tower Hospital from October 2007 to September 2009were prospectively studied.Of the 68 patients,30 received precise hemihepatectomy guided by MHV(precise group)and 38 received anatomical hemihepatectomy(traditional group).The types of hepatic vein in the precise group were evaluated and classified preoperatively.The operation time,operative blood loss,volume of blood transfusion,liver function,morbidity and length of hospital stay of the 2 groups were compared.All data were analyzed using the t test,rank sum test,chi-square test and Fisher exact probability.Results According to the Nakamura's classification,there were 17(57%)patients with type Ⅰ,8(27%)with type Ⅱ and 5(16%)with type Ⅲ;according to the Kawasaki's classification,there were 11 patients with type Ⅰ(37%)and 19 with type Ⅱ(63%).There were 13 patients received right hepatectomy with MHV preservation,15 received left hepatectomy with MHV preservation,1 received right hepatectomy without MHV preservation and 1 received left hepatectomy without MHV preservation.There were no significant difference in the volume of operative blood loss and blood transfusion,level of alanine transaminase,total bilirubin,cholinesterase at postoperative day 3,total length of hospital stay and length of postoperative hospital stay between the 2 groups(t = 1.07,0.92,0.07,0.21,0.63,0.63,0.75,P ＞ 0.05).The operation time,levels of albumin at postoperative day 3 and complication rate were (342 ± 113)minutes,(35 ±3)g/L and 40%(12/30)in the precise group,and(270 ±73)minutes,(33 ±3)g/Land 66%(25/38)in the traditional group,respectively,with significant differences between the 2 groups(t =2.79,2.19;x2 =4.49,P＜0.05).The positive rates of the resection margin were 5%(1/19)in the precise group and 35%(8/23)in the traditional group,there was a significant difference between the 2 groups(P ＜0.05).ConclusionPreoperative hepatic vein evaluation and precise hemihepateetomy guided by MHV can preserve the functional liver tissues with venous drainage,achieve adequate tumor-free resection margin and reduce the postoperative complication rate.

Objective:On the basis of the use of chemical methods to establish mouse model of lupus nephritis and its biological identification , we investigate the reverse effect of pathological lesions of B 7-1 human-mouse chimeric antibody blockade against B7/D28 signaling pathway in mice with lupus nephritis model.Methods:Pristane was injected intraperitoneally to 6-week-old female C57BL/6J mice at dose of 0.5 ml per mouse in one go,and urine protein,ANA and renal pathological changes were detected on a monthly basis.Mice whose urine protein content reached ++and ANA fluorescence intensity reached ++were randomly devided into three groups ,five each.Antibody intervention group was sequentially injected with B 7-1-mouse chimeric antibody by orbital venous , positive control group was injected with immunosuppressant CTX , negative control group was injected with isotype control IgG.Urine protein and ANA were also detected on a monthly basis.Mice were sacrificed three months after intervention was executed.Kidney was used for H&E dying , IC detection and electric microscope observation.Results: After four-month Pristane induction , urine protein content of 80%mice reached +-+++,meanwhile,serum ANA fluorescence intensity reached ++-+++.Glomerulonephritis infiltrating cells were observed Mice with urine protein and ANA , glomerular inflammatory cell infiltration , tubular epithelial cell degeneration visible edema ,vascular congestion significantly ,fibrosis.After antibody intervention ,urine protein content in antibody intervention group gradually reduced from ++-+++to ±-+++,ANA ++-+++to +-++,and were significantly different from that in the negative control group ( P<0.01 ).Analysis of kidney H&E dying showed that antibody glomerular infiltration of inflammatory cells in the intervention group and tubular congestion and other symptoms were improved significantly.Immunofluorescence staining indicated that fluorescence intensity of IC was significantly reduced in the antibody intervention group.Electron dense deposits reduction and glomerular basement membrane uniformity were observed in antibody intervention group by electric microscope when compared with the negative control group.Conclusion:B7-1 antibodies could downregulate immune response through inhibiting B 7-1/CD28 signaling pathway , reducing the production of autoantibodies and reversing pathological damage caused by autoimmune response .

A method for qualitative analysis of constituents in Panax notoginseng by UPLC-LTQ-Orbitrap mass spectrometry was established. Based on the high-resolution mass information, MS/MS fragmentation behaviors and chemical components from literatures, 43 compounds were identified or tentatively characterized. New type saponin aglycone, combined with malonyl-substituted and acetyl-substituted saponins were discovered and plausibly identified in this study. This work could be helpful for the quality control and further phytochemical studies of Panax notoginseng, and provided a good example for the analysis of chemical constituents in traditional Chinese medicine.

Objective To investigate the therapeutic effects of nano-hydroxyapatite/polyamide 66/platelet-rich plasma com-pound(n-HA/PA66/PRP)on the recovery of rabbit femur bone defect.Methods 40 New Zealand rabbits were artificially made to be bone defect by resecting the 1 cm substantia ossea with periosteum of femur,and were divided into two groups averagely depen-ding on implanted materials:experimental group(n-HA/PA66/PRP),control group (n-HA/PA66).Every five rabbits were sacri-ficed on week 2,4,8,12,and the femur healing status was observed by X ray,histology,and immunohistochemistry.Results No rabbit was infected or died,no implantation objects dropped.Gross observation,X-ray result and histology results demonstrated that the experimental group began to have a new bone tissue at 2 weeks after the operation,with the extension of time,the experimental group new bone growth speed and the quantity was better than the control group.The Lane-Sandhu method X-ray score showed that the experimental group (6.80±2.05)points and the control group(4.20±1.30)points at 12 weeks after the operation,and there were significant differences between two groups(P <0.05).Immunohistochemistry indicated that expression intensity of Vas-cular endothelial growth factor at 2 weeks and 4 weeks were significant differences between two groups(P <0.05),but there was no significant difference at 8 weeks and 12 weeks (P >0.05).Conclusion PRP combined with n-HA/PA66 artificial bone could accel-erate the healing of bone defect,and the effect of repair of bone defect is better than that of n-HA/PA66 artificial bone in the early.

Objective To explore the effect of Numb on tubular epithelial-to-mesenchymal transition (EMT) in rat proximal epithelial cells. Methods NRK52E cells were treated with different concentrations of recombinant human transforming growth factor-β1 (TGF-β1) (0, 1, 5, 10, 15, 20 μg/L) for 48 h or 10 μg/L TGF-β1 for different times (0, 24, 48, 72 h) in vitro. The expressions of E-cadherin, a-smooth muscle actin(α-SMA) and Numb in NRK 52E cells were detected by RT-PCR, Western blot and immunofluorescence staining. Meanwhile Numb siRNA oligo was transfected into NRK 52E cells with lipofectamine before TGF-β1 treatment, then Western blot was applied to detect the protein expression of E-cadherin, α-SMA and Numb in NRK52E cells. Results TGF-β1 could induce EMT in NRK52E cells in dose- and time-dependent manner. During the progress of TGF-β1-induced EMT, the protein expression of Numb in 5, 10, 15, 20 μg/L group was 1.33 folds (P=0.024), 1.39 folds (P=0.035), 1.45 folds (P=0.025), 1.51 folds (P=0.000) respectively as compared to 0 μg/L group. Likewise, the protein and mRNA expression of Numb in 24 h, 48 h, 72 h group was 1.48 folds (P=0.046) and 1.56 folds (P=0.012), 1.54 folds (P=0.011) and 1.82 folds (P=0.008), 1.79 folds (P=0.028) and 1.82 folds (P=0.002) respectively as compared to 0 h group. Moreover, large amount of Numb was accumulated in the cytoplasm. Down-regulation of Numb expression by siRNA transfection did not influence the basal expression of E-cadherin and α-SMA in NRK 52E cells, but attenuated the progression of EMT in NRK52E cells induced by TGF-β1. The up-regulation of α-SMA protein was reduced to 18.1% (P=0.004) while the down-regulation of E-cadherin protein was reversed to 2.19 folds (P=0.004). Conclusion Numb can promote EMT in rat proximal epithelial cells.

Objective To investigate the immunoregulatory effects of bone marrow-derived mesenehy-real stem ceils (BMSCs) combined with leflunomide (LEF) on mice T-lymphocytes in vitro. Methods BMSCs from BALB/c mice were isolated and expanded. The purity of BMSCs was identified by flow cytometry (FCM). The BALB/c mice's spleen lymphocytes were isolated by using EZ-Sep<'TM> Mouse 1X. Under ConA stimulation, spleen lymphocytes were pretreated with LEF, then washed and co-cuhured with BMSCs. We set up four groups to investigate in this study: group A, spleen lymphocytes alone; group B, spleen lymphocytes with BM- SCs; group C, LEF-pretreated spleen lymphocytes alone and the group D, LEF-pretreated spleen lymphocytes with BMSCs. T-lymphocytes proliferation was assessed by MTT. FCM was used to analysis T-lymphocytes apoptosis and surface markers of CD69 and CD28. The mRNA expression of interleukin (IL)-2, IL-10 were detected by real-time RT-PCR. Results In vitro, the T-lymphocytes'values of A570 nm were significantly lower in group B and group C, compared with group A (group B vs group C vs group A, 0.578±0.042 vs 0.502± 0.040 vs 0.778±0.035, P<0.01), while the value of A<,570 nm>in group D was 0.218±0.033, which was also obviously lower than that in group B and group C (P<0.01). There were no suppressing effects on T-lympho-cytes'activation and expression of IL-2 had been observed. The proportion of apoptotic T-lymphocytes in group B and group D were (2.29±0.32)％ and (4.22±0.98)％, which was significandy lower than that in group A (8.08±1.20) (P<0.01). The expression of IL-10 in group B and C were also lower than that in group A (group B vs group C vs group A, 0.098±0.039 vs 0.054±0.022 vs 1.000, P<0.01 ). Either, the expression of IL-10 in group D was 0.023±0.015, which was obviously lower than that in group B and group C (P<0.01). Conclusion BMSCs combined with LEF show synergistic immunoregulatary effects on mice's T-lymphoeytes.

Objective To investigate the applicability of Cys C-based formulas for prediction of GFR in Chinese patients with CKD.Methods A total of 176 adult patients with CKD including 90 males and 86 females collected from 4 hospitals located in different geographic regions of China (Beijing,Shanghai,Dalian and Changsha) were enrolled in this study from September 2007 to July 2009.The rGFR was measured using 99mTc-DTPA clearance rate two-sample method.Cystantic C was measured by PETIA and PENIA respectively.The results of eGFR in the Larsson formula,Grubb formula,Hoek formula,Filler formula,Stevens formula and Hojs formula were compared with the rGFR to evaluate the calculation coherence,bias,precision,accuracy and the performance of correct phasing of the formulas.Results The mean 99mTc-DTPA clearance was [40.70 ( 19.09 - 86.49)] ml · min-1 · ( 1.73m2 ) -1.Significant difference was witnessed in the evaluation of GFR estimation formulas calculated by PETIA and PENIA (P ＜0.01).ICC and Spearman correlation analysis revealed a significant correlation between eGFR and rGFR.The ICCs of eGFR and rGFR ranged from 0.874 to 0.938.Compared with rGFR,the 30％ accuracy of all the eight evaluation formulas using PETIA and PENIA method were lower than 60％.The percentages of correct phasing in all the 5 stages of CKD were not ideal.With these formulas,percentages of correct phasing from CKD stage 2 to CKD stage 4 were lower than 65％.The eGFRs were underestimated by formulas evaluated by PENIA in CKD stage 1.All the eGFRs were overestimated remarkably by all equations in CKD stage 5.Conclusions None of the eight Cys C based formulas are ideal for estimation of GFR in Chinese CKD patients,and they can not be applied to Chinese patients directly.For this patient population,further studies will be needed to develop a more accurate Cys C-based GFR estimation formula that includes ethnicity,age and other factors.