1. Volume 17, Issue 1, Pages 42-50 (January 2009)
Nonviral Retrograde Gene Transfer of Human Hepatocyte Growth Factor Improves Neuropathic Pain-related Phenomena in Rats 
Toyokazu Tsuchihara, Sho Ogata, Koichi Nemoto, Takatoshi Okabayashi, Kuniaki Nakanishi, Naoki Kato, Ryuichi Morishita, Yasufumi Kaneda, Maki Uenoyama, Shinya Suzuki, Masatoshi Amako, Toshiaki Kawai, Hiroshi Arino 
Molecular Therapy 
Volume 17, Issue 1, Pages (January 2009)
DOI: /mt
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

2. Figure 1
Analysis of human hepatocyte growth factor (HGF)-like immunoreactive protein, and endogenous rat HGF mRNA and protein. (a) Expression of human HGF-like immunoreactive protein was detected in ipsilateral sciatic nerve, dorsal root ganglia (DRG), and spinal cord at the 3-day, 7-day, and 10-day time points in the CCI+HGF (HGF gene-transfer after chronic constriction injury (CCI)) group (red bars), but not in the CCI+CV (control-vector gene-transfer after CCI) group (yellow bars) nor in the CCI (green bars) or non-CCI (aged-matched non-CCI controls) (blue bars) groups. (b) Reverse transcriptase-PCR for rat HGF mRNA in rat ipsilateral sciatic nerve and human liver. Expression of rat HGF mRNA was detected in rat nerve, but not in human liver. (c) Expression of endogenous rat HGF mRNA in the ipsilateral sciatic nerve and DRG in four groups (CCI+HGF (red bars), CCI+CV (yellow bars), CCI (green bars), and non-CCI (blue bars) groups). (d) Expression of endogenous rat HGF-like immunoreactive protein in the ipsilateral sciatic nerve and DRG in four groups (CCI+HGF (red bars), CCI+CV (yellow bars), CCI (green bars), and non-CCI (blue bars) groups). (e) Immunohistochemistry for human HGF. Human HGF-like immunoreactive protein was detected in the ipsilateral muscle, sciatic nerve, DRG, and spinal cord dorsal horn neurons (arrowheads) in the CCI+HGF group on day 3 after both the first and second gene transfers, but not in any of these tissues in the CCI+CV group. ND, not detected; arrows (in a, c, and d), gene transfer; bar = 20 µm; *P < 0.05; **P < 0.01; (n = 6 for each group); #not done (DRG at 6 weeks in c).
Molecular Therapy  , 42-50DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

3. Figure 2
Behavioral studies. (a) Time course of changes in the tactile-response threshold in the CCI (chronic constriction injury) group (black triangle), CCI+HGF (HGF gene-transfer after chronic constriction injury (CCI)) group (black circle), and CCI+CV (control-vector gene-transfer after CCI) group (gray square). (b) Time course of changes in the withdrawal latency in the above three groups. Arrows, gene transfer; **P < 0.01 versus CCI and CCI+CV groups (n = 16 for each group).
Molecular Therapy  , 42-50DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

4. Figure 3
Laser Doppler flux and thermographic studies. (a) Time course of changes in laser Doppler flux in the sciatic nerve of the CCI (chronic constriction injury) group (black triangle), CCI+HGF (HGF gene-transfer after chronic constriction injury (CCI)) group (black circle), and CCI+CV (control-vector gene-transfer after CCI) group (gray square). (b) Time course of changes in laser Doppler flux in the hind paw, together with fluxmetry scanning photographs taken at 3 weeks after the first gene transfer in the above three groups. (c) Time course of changes in the temperature of the hind paw (circled in accompanying temperature photographs taken at 1 week after the first gene transfer) in the above three groups. Arrows, gene transfer; **P < 0.01 versus CCI and CCI+CV groups (n = 6 for each group). #P < 0.05 versus preoperative value (“week −1”).
Molecular Therapy  , 42-50DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

5. Figure 4
Histological studies of the sciatic nerve. (a) Semi-thin transverse sections, size-frequency distributions for myelinated axons, and number of large myelinated axons (>5 µm) observed in a mm2 area of the sciatic nerve at 3 weeks after the first gene transfer in the CCI (chronic constriction injury), CCI+CV (control-vector gene-transfer after CCI), CCI+HGF (HGF gene-transfer after chronic constriction injury (CCI)), and non-CCI groups. Degeneration of wallerian-type fibers (arrowheads) was sometimes observed in the CCI and CCI+CV groups. Bar = 20 µm; *P < 0.05; **P < 0.01 (n = 6 for each group). (b) Representative electron microscopic sections, size-frequency distributions, and mean diameter of unmyelinated axons in the sciatic nerve at 3 weeks after the first gene transfer. Bar = 2 µm; **P < 0.01 (n = 6 for each group).
Molecular Therapy  , 42-50DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

6. Figure 5
RT-PCR studies in dorsal root ganglia (DRG) and sciatic nerve. (a) P2X2, P2X3, P2X4, and P2Y1 receptors (n = 6 for each group). (b) Interleukin-6 (IL-6) (n = 5 for each group). (c) Activating transcription factor 3 (ATF3) (n = 5 for each group). Observations were made at 1 and 3 weeks after the first gene transfer in the CCI (chronic constriction injury), CCI+CV (control-vector gene-transfer after CCI), CCI+HGF (HGF gene-transfer after CCI), and non-CCI (aged-matched non-CCI) groups. *P < 0.05.
Molecular Therapy  , 42-50DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions