A Non-immunogenic Bivalent d-Protein Potently Inhibits Retinal Vascularization and Tumor Growth.

We report a general approach to engineering multivalent d-proteins with antibody-like activities in vivo. Mirror-image phage display and structure-guided design were utilized to create a d-protein that uses receptor mimicry to antagonize vascular endothelial growth factor A (VEGF-A). Selections against the d-protein form of VEGF-A using phage-displayed libraries of two different domain scaffolds yielded two proteins that bound distinct receptor interaction sites on VEGF-A. X-ray crystal structures of the d-protein/VEGF-A complexes were used to guide affinity maturation and to construct a heterodimeric d-protein VEGF-A antagonist with picomolar activity. The d-protein VEGF-A antagonist prevented vascular leakage in a rabbit eye model of wet age-related macular degeneration and slowed tumor growth in the MC38 syngeneic mouse tumor model with efficacies comparable to those of approved antibody drugs, and in contrast with antibodies, the d-protein was non-immunogenic during treatment and following subcutaneous immunizations.

[Effects of Qufengtongluo Recipe on podocin mRNA expression and podocyte morphology in rats with adriamycin-induced nephropathy].

OBJECTIVE: To observe the effects of Qufengtongluo Recipe (QFTLR) on the expression of podocin mRNA and podocyte morphology in rats with adriamycin-induced nephropathy (AN), and explore the possible mechanism mediating the therapeutic effect of QFTLR on nephropathic proteinuria. METHODS: SD rats were randomized into normal control group, AN model group (established by a single injection of adriamycin via the tail vein), and 3 intervention groups with QFTLR, prednisone, or benazepril treatment. After the corresponding treatments, the expression of podocin mRNA in the renal tissues was detected by RT-PCR methods, and the morphological changes of the podocytes were examined by electron microscope. RESULTS: Compared with the normal control group, the AN model group showed significantly lowered expressions of podocin mRNA (P<0.01) with reduced podocytes and widening, fusion or even absence of the foot processes (FP). Intervention with QFTLR significantly increased the expression of podocin mRNA (P<0.01) and the number of podocytes, and obviously lessened the structural changes of the FP. CONCLUSION: QFTLR can produce therapeutic effect against nephropathic proteinuria possibly by up-regulating the expression of podocin mRNA and improving the morphological changes of the podocytes.

[Effects of qufengtongluo recipe on expressions of HSPG mRNA and protein in adriamycin-induced nephropathy rats].

OBJECTIVE: To investigate the expression of HSPG in glomerular base membrane of adriamycin-induced nephropathy (AN) rats, and the effect of Qufengtongluo recipe on HSPG mRNA expression and proteinuria in AN rats. METHODS: One hundred forty rats were used in this study, including 32 rats in normal control group. AN was induced in the left rats by a single tail intravenous injection of adriamycin. Three weeks later, 90 AN rats were randomly divided into five groups; the nephropathy group (B, n=18), the Qufeng group (C, n=18), Qufeng and prednisone group (D, n=18), prednisone group(E,n=18) and benazepri group (F, n= 18). The rats in these five groups were treated with different combination of Qufeng recipe and prednisone. In each group, renal tissue samples were collected at week 3 and 7. The distribution, expression of HSPG was examined by indirect immunofluorescence, and semi-quantity RT-PCR, respectively. RESULTS: (1) In AN rats, the diffuse fusion and effacement of foot processes were observed when model established. (2) Compared with nephropathy group, the average fluorescence intensity of HSPG dramatically increased in Qufeng group and prednisone group (P < 0.01), similarly, it also increased in D and F groups (P < 0.01). (3) Compared with nephropathy group, the expression of HSPG mRNA was significantly up-regulated in other groups. (P < 0.01), especially in C and F groups. There was significant negative correlation between the expression of HSPG and quantity of 24-hour proteinuria. CONCLUSION: The abnormal expression of HSPG and their altered distributions may be an important molecular mechanism that leads to the occurrence and development of proteinuria in AN rats. The effect of Qufengtongluo recipe on nephrotic syndrome might be related to the alteration of HSPG expression and distribution in glomerulus.

[Effect of Qufengtongluo recipe on expression of nephrin mRNA in adriamycin-induced nephropathy in rats].

OBJECTIVE: To investigate the expression of nephrin mRNA in adriamycin-induced nephropathy (AN) in rats, and to explore the effect of Qufengtongluo recipe on proteinuria in AN rats and on the expression of nephrin mRNA. METHODS: Adriamycin nephropathy was induced by a single tail intravenous injection of adriamycin. We randomly divided 140 rats into a normal control group (n=32) and a nephropathy model group (n=108). Three weeks later, 90 AN rats were randomly divided into 5 groups: a model group, a qufeng group, a qufeng and prednisone group, a prednisone group, and a benazepri group (18 rats in each group). They were treated respectively, and renal tissue samples were collected at week 0, 3, 5, and 7, respectively. The distribution and expression of nephrin mRNA were examined by indirect immunofluorescence and semi-quantity RT-PCR. RESULTS: In the AN rats, the diffuse fusion and effacement of foot processes were observed at week 3. The fluorescence intensity of nephrin and the expression of nephrin mRNA significantly increased in the qufeng group and the prednisone group compared with the model group at the week 7 (P<0.01). CONCLUSION: Abnormal expression of nephrin is the important molecular mechanism that leads to the occurrence and development of proteinuria in AN rats. Qufengtongluo recipe has effect on nephrotic syndrome through altering the expression and distribution of nephrin in glomerulus.

[Clinical and pathological study on effects of Qianggan Capsule combined lamivudine on hepatic fibrosis in patients with chronic hepatitis B].

OBJECTIVE: To study the therapeutic effects of Qianggan Capsule (QC) combined Lamivudine on hepatic fibrosis in patients with chronic hepatitis B. METHODS: Eighty-five patients were randomly divided into two groups, group A (40 cases) were treated with QC and Lamivudine and group B (45 cases) were treated with QC alone both for 6 months. Hepatic fibrosis related indexes and pathologic examination of liver biopsy were performed within 3 months before treatment and in 1 month after treatment. RESULTS: Serum levels of hyaluronic acid, collagen N and laminin decreased markedly after treatment in both groups (P < 0.05). Hepatic histopathological examination showed that the total effective rate of impovement in activity of inflammation-necrosis and fibrosis was 80.0% and 70.0% in group A, 57.8% and 75.6% in group B, respectively, the combined treatment showed a better effect in improving the activity of inflammation-necrosis than QC alone (P < 0.05), but with no significant difference to the latter in improving fibrosis. CONCLUSION: QC combined Lamivudine could markedly reduce the activity of hepatic inflammation-necrosis, QC alone could also improve hepatic fibrosis.