The Effects of Nerve Growth Factor on Adult and Aged Dorsal Root Ganglion Neurons Maintained in Primary Culture

Nerve growth factor (NGF) is, a prototype neurotrophic factor, well documented for its biochemical character, distribution and functions in embryonic and neonatal animals. However, only very tentative evidence exists for its effects on mature neurons, and results are controversial. Studying NGF by using mature neurons should be a significant and attractive field, not only for its clinical value in nerve regeneration by adults, but also for understanding the mechanisms of aging. Studies in vitro present a comparatively simple environment for analysis of the effective factors. Dorsal root ganglia (DRGs), containing an enormous heterogeneity of sensory neuronal types, are a useful model in understanding the nervous system. Hence, in the present study, mouse DRG neurons were maintained in vitro supplemented with NGF V5 cultures without NGF or with anti-NGF present, to examine any NGF effects on survival, morphological phenotype, neuropeptide expressions and neurite outgrowth (equivalent to regeneration in vitro). Comparison of adult (6 months) and aged (2 years) animals was made throughout the present study. In the survival study, a range of NGF (25-200ng/ml) was added to adult neurons in co-cultures, and neuronal survival monitored for 14 days in vitro (div). 100ng/ml NGF effectively maintained survival, compared with cultures without exogenous NGF. Aged as well as adult neurons were cultured [with non-neuronal cells (NNCs) present] without exogenous NGF or with 100ng/ml NGF added for up to 29div. NGF enhanced the survival of both adult and aged neurons (P<0.005 by ANOVA). To exclude possible endogenous NGF from NNCs, or any mediation of an NGF effect in co-culture by NNCs , a neuron- enriched culture system was also used. In these neuronal loss was avoided during cell preparation and well dispersed neurons were obtained. 100ng/ml NGF and/or 1:100 anti-NGF were re-examined in the modified neuron-enriched cultures for 9div and an enhanced survival of both adult and aged neuron persisted, although this effect was lower than in co-cultures. In general, about 20% of DRG neurons were NGF-dependent for their survival in adult and aged cultures. By size analysis of over 6500 neurons, intermediate-sized neurons (24-33mum in diameter) were predominant for both adult and aged in co-culture in the presence of NGF. The immunocytochemistiy (ICC) study using the avidin-biotin- peroxidase complex (ABC) method revealed that higher proportions of SP- immunoreactive (ir), CGRP-ir and NPY-ir aged, as well as adult, neurons were present in enriched cultures supplemented with 10 or 100ng/ml NGF than in cultures without NGF added (P<0.05-0.01 by ANOVA and t-tests), whereas SOM-ir adult and aged neurons showed little difference in cultures without or with NGF added. By counting at the peak day (9div) of SP expression, enhancement of the SP-ir subset in the presence of vs absence of NGF was similar for adult or aged neurons; the proportion of the SP-ir subset was not reduced following aging. Hence the SP-ir subset, together with the subsets of CGRP-ir and NPY-ir, were considered as NGF-dependent survival neurons in adult, and even aged mouse DRGs. Enhancement of the SP-ir proportion was more rapid in aged neuronal cultures supplemented with NGF than in cultures without exogenous NGF. Also, the staining intensities for CGRP and NPY were greater in both adult and aged cultures with NGF added than those in cultures without NGF. In addition to maintaining survival, NGF affected the mature neuronal phenotypes. Scanning election microscopy demonstrated that in the presence of NGF, abundant microvilli projections were distributed on the neuronal surface; in contrast, neurons were deformed and lost their smooth appearance, if 1:100 anti-NGF was present in enriched cultures. In the study of neurite geometry in enriched culture, major neurite length (maximal extension of a neurite), entire neurite length (lengths of major neurite plus all its branches), total neurite length (lengths of all neurite outgrowth from each neuron), soma size, neurite number (for each neuron) and branch number (per neurite) were selected as parameters. Tracing was carried out blindly at 1, 3, 6 and 9div by computer image analysis and a digitizing tablet. Neurites and neurons from at least 5 mice for each adult or aged cultures were measured directly from culture or from photomicrograph montages and data were statistically tested by ANOVA. The results demonstrated that NGF enhanced neurite outgrowth (P<0.0005) at 6 and 9div for both adult and aged neurons Total neurite lengths of adult and aged neurons were distinct in the presence of NGF; neurite length was predominantly enhanced by adult neurons, whereas increased branch lengths were evident for aged neurons. Neurites predominantly extended from intermediate- and large-sized neurons. In summary, aged neurons showed comparatively lower abilities for survival and functional peptide expression compared to adult neurons in vitro, but continued to be responsive to exogenous NGF. In addition, reconstruction of neurites by aged and adult neurons in vitro was demonstrated in the present study

Excited Heavy Quarkonium Production at the LHC through WW-Boson Decays

Sizable amount of heavy-quarkonium events can be produced through $W$-boson decays at the LHC. Such channels will provide a suitable platform to study the heavy-quarkonium properties. The "improved trace technology", which disposes the amplitude ${\cal M}$ at the amplitude-level, is helpful for deriving compact analytical results for complex processes. As an important new application, in addition to the production of the lower-level Fock states $|(Q\bar{Q'})[1S]>$ and $|(Q\bar{Q'})[1P]>$, we make a further study on the production of higher-excited $|(Q\bar{Q'})>$-quarkonium Fock states $|(Q\bar{Q'})[2S]>$, $|(Q\bar{Q'})[3S]>$ and $|(Q\bar{Q'})[2P]>$. Here $|(Q\bar{Q'})>$ stands for the $|(c\bar{c})>$-charmonium, $|(c\bar{b})>$-quarkonium and $|(b\bar{b})>$-bottomonium respectively. We show that sizable amount of events for those higher-excited states can also be produced at the LHC. Therefore, we need to take them into consideration for a sound estimation.Comment: 7 pages, 9 figures and 6 tables. Typo errors are corrected, more discussions and two new figures have been adde

Short-Term Efficacy of Laparoscopic Treatment for Colorectal Cancer in Patients with Schistosomiasis Japonica

Introduction. Schistosomiasis is associated with numerous complications such as thrombocytopenia, liver cirrhosis, portal hypertension, and colitis. To the best of our knowledge, the feasibility and outcomes of laparoscopic colorectal surgery in patients with schistosomiasis have not yet been studied. Methods. In this study, the data of 280 patients with colorectal carcinoma along with schistosomiasis japonica infection who underwent laparoscopic or open colorectal surgery were retrospectively analyzed. Preoperative data, operative data, pathological outcomes, postoperative complications, and recovery were compared between patients in the laparoscopic (LAC) and open (OC) groups. Results. There were no significant differences in the preoperative data between the groups. However, fewer postoperative complications, especially severe hypoproteinemia, early postoperative feeding, and shorter postoperative hospital stay, were observed in patients in the LAC group (P<0.001). The mean operative time was higher in the LAC group (180 min versus 158 min; P<0.001), while the mean blood loss was similar (95 mL versus 108 mL; P=0.196) between groups. The mean number of lymph nodes harvested was also similar in both groups (15 versus 16; P=0.133). Conclusion. Laparoscopic surgery for colorectal cancer is safe in patients with schistosomiasis japonica and has better short-term outcomes than open surgery

Neuroprotective effects of bis(7)-tacrine against glutamate-induced retinal ganglion cells damage

<p>Abstract</p> <p>Background</p> <p>Glutamate-mediated excitotoxicity, primarily through N-methyl-D-aspartate (NMDA) receptors, may be an important cause of retinal ganglion cells (RGCs) death in glaucoma and several other retinal diseases. Bis(7)-tacrine is a noncompetitive NMDA receptors antagonist that can prevent glutamate-induced hippocampal neurons damage. We tested the effects of bis(7)-tacrine against glutamate-induced rat RGCs damage in vitro and in vivo.</p> <p>Results</p> <p>In cultured neonatal rats RGCs, the MTT assay showed that glutamate induced a concentration- and time-dependent toxicity. Bis(7)-tacrine and memantine prevented glutamate-induced cell death in a concentration-dependent manner with IC50 values of 0.028 μM and 0.834 μM, respectively. The anti-apoptosis effects of bis(7)-tacrine were confirmed by annexin V-FITC/PI staining. In vivo, TUNEL analysis and retrograde labeling analysis found that pretreatment with bis(7)-tacrine(0.2 mg/kg) induced a significant neuroprotective effect against glutamate-induced RGCs damage.</p> <p>Conclusions</p> <p>Our results showed that bis(7)-tacrine had neuroprotective effects against glutamate-induced RGCs damage in vitro and in vivo, possibly through the drug's anti-NMDA receptor effects. These findings make bis(7)-tacrine potentially useful for treating a variety of ischemic or traumatic retinopathies inclusive of glaucoma.</p

Heavy Quarkonium Production at LHC through WW Boson Decays

The production of the heavy $(c\bar{c})$-quarkonium, $(c\bar{b})$-quarkonium and $(b\bar{b})$-quarkonium states ($(Q\bar{Q'})$-quarkonium for short), via the $W^+$ semi-inclusive decays, has been systematically studied within the framework of the non-relativistic QCD. In addition to the two color-singlet $S$-wave states, we also discuss the production of the four color-singlet $P$-wave states $|(Q\bar{Q'})(^1P_1)_{\bf 1}>$ and $(Q\bar{Q'})(^3P_J)_{\bf 1}>$ (with $J=(1,2,3)$) together with the two color-octet components $|(Q\bar{Q'})(^1S_0)_{\bf 8}>$ and $|(Q\bar{Q'})(^3S_1)_{\bf 8}>$. Improved trace technology is adopted to derive the simplified analytic expressions at the amplitude level, which shall be useful for dealing with the following cascade decay channels. At the LHC with the luminosity ${\cal L}\propto 10^{34}cm^{-2}s^{-1}$ and the center-of-mass energy $\sqrt{S}=14$ TeV, sizable heavy-quarkonium events can be produced through the $W^+$ boson decays, i.e. $2.57\times10^6$ $\eta_c$, $2.65\times10^6$ $J/\Psi$ and $2.40\times10^6$ $P$-wave charmonium events per year can be obtained; and $1.01\times10^5$ $B_c$, $9.11\times10^4$ $B^*_c$ and $3.16\times10^4$ $P$-wave $(c\bar{b})$-quarkonium events per year can be obtained. Main theoretical uncertainties have also been discussed. By adding the uncertainties caused by the quark masses in quadrature, we obtain $\Gamma_{W^+\to (c\bar{c})+c\bar{s}} =524.8^{+396.3}_{-258.4}$ KeV, $\Gamma_{W^+\to (c\bar{b})+b\bar{s}} =13.5^{+4.73}_{-3.29}$ KeV, $\Gamma_{W^+\to (c\bar{b})+c\bar{c}}= 1.74^{+1.98}_{-0.73}$ KeV and $\Gamma_{W^+\to (b\bar{b})+c\bar{b}}= 38.6^{+13.4}_{-9.69}$ eV.Comment: 24 pages, 12 figures. References updated. To be published in Phys.Rev. D. To match the published versio