Gene therapy of Parkinson's disease using adeno-associated virus (AAV) vectors

Parkinson's disease (PD) is characterized by the progressive loss of the dopaminergic neurons in the substantia nigra and a severe decrease in dopamine in the striatum. A promising approach to the gene therapy of PD is intrastriatal expression of dopamine-synthesizing enzymes [tyrosine hydroxylase (TH) and aromatic L-amino acid decarboxylase (AADC)]. The most appropriate gene-delivery vehicles for neurons are adeno-associated virus (AAV) vectors, which are derived from non-pathogenic virus. Therefore, TH and AADC genes were introduced into the striatum in the lesioned side using separate AAV vectors in parkinsonian rats, and the coexpression of TH and AADC resulted in better behavioral recovery compared with TH alone. Another strategy for gene therapy of PD is the protection of dopaminergic neurons in the substantia nigra using an AAV vector containing a glial cell line-derived neurotrophic factor (GDNF) gene. Combination of dopamine-supplement gene therapy and GDNF gene therapy would be a logical approach to the treatment of PD.     

The V264M polymorphism of tissue factor pathway inhibitor gene in patients with acute coronary syndrome

Objective： To investigate V264M polymorphism of tissue factor pathway inhibitor （TFPI） in patients with acute coronary syndrome （ACS） in Chinese. Methods ： The genotypes of V264M were detected by polymerase chain reaction （PCR） and restriction fragment length polymorphism （PCR-RFLP） in 136 patients with ACS and 106 healthy controls with their plasmal f-TFPI tested by enzyme linked immunosorbent assay （ELISA）. Results： Two genotypes were found in V264M： GG and GA. The distribution frequencies of alleles and genotypes were in accordance with those predicted by Hardy Weinberg equilibrium in the present study （χ2 = 0.437, P 〉 0.05）. Plasma f-TFPI level was lower in A allele bearer than that in non-A allele carriers and higher in ACS than control subjects （P〈0.05）. No relationship was found between ACS and V264M polymorphism （P〉0.05）. Conclusion： The V264M polymorphism may have an impact on the plasma level of free TFPI（f-TFPI）, but it has no relationship with ACS.     

Technical refinement in adult-to-adult living donor liver transplantation using right lobe graft

OBJECTIVE: To report the authors' experience with living donor liver transplantation in adults using right lobe liver grafts, performed by a modified technique. SUMMARY BACKGROUND DATA: The initial results of seven living donor liver transplants in adults using extended right lobe grafts were satisfactory, but serious complications occurred in two donors, and six recipients required repeat laparotomy. Another 11 similar operations were performed. Further evaluation was made with the aim of improving the postoperative outcome. METHODS: From December 1996 to August 1998, 11 patients underwent living donor liver transplantation using right lobe grafts. The first four patients underwent surgery using methods previously designed and the next seven underwent a modification designed to minimize devitalized tissues on the liver transection surface, improve hepatic venous drainage, and reduce the number of hepatic duct orifices. RESULTS: There were no donor deaths. Donor complications included cholestasis (n = 1) and minor wound infection (n = 1). All the first four recipients required a repeat laparotomy for infected necrotic liver transection surface (n = 1), acute pancreatitis (n = 1), hepatic vein thrombosis (n = 1), and leakage from one of the two bilioenteric anastomoses (n = 1). The patient with hepatic vein thrombosis died. In the last seven recipients, all of whom survived the operation, one required a repeat laparotomy with the discovery of a methicillin-resistant Staphylococcus aureus culture of fibrinous exudate at the left subphrenic peritoneum, and another had right hepatic duct stump necrosis. The latter was likely related to hypovolemic shock secondary to bleeding from the right saphenous vein on removal of a hemofiltration catheter. Comparison of the incidence of repeat laparotomy between the first four and the remaining seven recipients showed a significant trend of improvement. Combining the result of the seven patients reported previously, the improvement in terms of relaparotomy rate is significant. CONCLUSION: With modification of surgical technique, living donor liver transplantation in adults using right lobe liver grafts can become a relatively safe procedure.     

Gatifloxacin inducing apoptosis of stromal fibroblasts through cross-talk between caspase-dependent extrinsic and intrinsic pathways

AIM: To reveal the cytotoxicity and related mechanisms of gatifloxacin (GFX) to stromal fibroblasts (SFs) in vitro. METHODS: SFs were treated with GFX at different concentrations (0.009375%-0.3%), and their viability was detected by MTT method. The cell morphology was observed using light/transmission electron microscope. The plasma membrane permeability was measured by AO/EB double-staining. Then cell cycle, phosphatidylserine (PS) externalization, and mitochondrial transmembrane potential (MTP) were analyzed by flow cytometry. DNA damage was analyzed by electrophoresis and immunostaining. ELISA was used to evaluate the caspase-3/-8/-9 activation. Finally, Western blotting was applied for detecting the expressions of apoptosis-related proteins. RESULTS: Morphological changes and reduced viability of GFX-treated SFs demonstrated that GFX above 0.009375% had cytotoxicity to SFs with dependence of concentration and time. GFX-treating cells also showed G1 phase arrest, increased membrane permeability, PS externalization and DNA damage, which indicated that GFX induced apoptosis of SFs. Additionally, GFX could activate the caspase-8, caspase-9, and caspase-3, induce MTP disruption, downregulate B-cell leukemia-2 (Bcl-2) and B-cell leukemia-XL (Bcl-XL), and upregulate Bcl-2 assaciated X protein (Bax), Bcl-2-associated death promoter (Bad), Bcl-2 interacting domain (Bid) and cytoplasmic cytochrome C in SFs, suggesting that caspase-dependent extrinsic and intrinsic pathways were related to GFX-contributed apoptosis of SFs. CONCLUSION: The cytotoxicity of GFX induces apoptosis of SFs through triggering the caspase-dependent extrinsic and intrinsic pathways.     

Migration capacity of human umbilical cord mesenchymal stem cells towards glioma in vivo*

High-grade glioma is the most common malignant primary brain tumor in adults.The poor prognosis of glioma,combined with a resistance to currently available treatments,necessitates the development of more effective tumor-selective therapies.Stem cell-based therapies are emerging as novel cell-based delivery vehicle for therapeutic agents.In the present study,we successfully isolated human umbilical cord mesenchymal stem cells by explant culture.The human umbilical cord mesenchymal stem cells were adherent to plastic surfaces,expressed specific surface phenotypes of mesenchymal stem cells as demonstrated by flow cytometry,and possessed multi-differentiation potentials in permissive induction media in vitro.Furthermore,human umbilical cord mesenchymal stem cells demonstrated excellent glioma-specific targeting capacity in established rat glioma models after intratumoral injection or contralateral ventricular administration in vivo.The excellent glioma-specific targeting ability and extensive intratumoral distribution of human umbilical cord mesenchymal stem cells indicate that they may serve as a novel cellular vehicle for delivering therapeutic molecules in glioma therapy.     

Neuropeptide Y gene transfection inhibits post-epileptic hippocampal synaptic reconstruction

Exogenous neuropeptide Y has antiepileptic effects; however, the underlying mechanism and optimal administration method for neuropeptide Y are still unresolved. Previous studies have used intracerebroventricular injection of neuropeptide Y into animal models of epilepsy. In this study, a recombinant adeno-associated virus expression vector carrying the neuropeptide Y gene was injected into the lateral ventricle of rats, while the ipsilateral hippocampus was injected with kainic acid to establish the epileptic model. After transfection of neuropeptide Y gene, mossy fiber sprouting in the hippocampal CA3 region of epileptic rats was significantly suppressed, hippocampal synaptophysin (p38) mRNA and protein expression were inhibited, and epileptic seizures were reduced. These experimental findings indicate that a recombinant adeno-associated virus expression vector carrying the neuropeptide Y gene reduces mossy fiber sprouting and inhibits abnormal synaptophysin expression, thereby suppressing post-epileptic synaptic reconstruction.     

Ghrelin-ghrelin O-acyltransferase system in the pathogenesis of nonalcoholic fatty liver disease

Nonalcoholic fatty liver disease(NAFLD) is currently considered as the most common liver disease in Western countries,and is rapidly becoming a serious threat to public health worldwide.However,the underlying mechanisms leading to the development of NAFLD are still not fully understood.The ghrelin-ghrelin O-acyltransferase(GOAT) system has recently been found to play a crucial role in both the development of steatosis and its progression to nonalcoholic steatohepatitis.Ghrelin,the natural ligand of the growth hormone secretagogue receptor,is a 28-amino acid peptide possessing a unique acylation on the serine in position 3 catalyzed by GOAT.The ghrelin-GOAT system is involved in insulin resistance,lipid metabolism dysfunction,and inflammation,all of which play important roles in the pathogenesis of NAFLD.A better understanding of ghrelin-GOAT system biology led to the identification of its potential roles in NAFLD.Molecular targets modulating ghrelin-GOAT levels and the biologic effects are being studied,which provide a new insight into the pathogenesis of NAFLD.This review probes into the possible relationship between the ghrelin-GOAT system and NAFLD,and considers the potential mechanisms by which the ghrelin-GOAT system brings about insulin resistance and other aspects concerning NAFLD.     

Short-term environmental enrichment exposure induces proliferation and maturation of doublecortin-positive cells in the prefrontal cortex

Previous studies have demonstrated that doublecortin-positive immature neurons exist pre- dominantly in the superficial layer of the cerebral cortex of adult mammals such as guinea pigs, and these neurons exhibit very weak properties of self-proliferation during adulthood under physiological conditions. To verify whether environmental enrichment has an impact on the proliferation and maturation of these immature neurons in the prefrontal cortex of adult guinea pigs, healthy adult guinea pigs were subjected to short-term environmental enrichment. Animals were allowed to play with various cognitive and physical stimulating objects over a period of 2 weeks, twice per day, for 60 minutes each. Immunofluorescence staining results indicated that the number of doublecortin-positive cells in layer II of the prefrontal cortex was significantly increased after short-term environmental enrichment exposure. In addition, these doublecortin-positive cells co-expressed 5-bromo-2-deoxyuridine （a marker of cell prolifera- tion）, c-Fos （a marker of cell viability） and NeuN （a marker of mature neurons）. Experimental findings showed that short-term environmental enrichment can induce proliferation, activation and maturation of doublecortin-positive cells in layer II of the prefrontal cortex of adult guinea pigs.     

Analysis of the relationship between postoperative ophthalmic complications and dialysis time of pre-kidney transplantation

AIM: To determine the influence of the dialysis time before kidney transplantation on postoperative ophthalmic complications. METHODS: One hundred and eighty three patients who were given the follow-up after kidney transplantation were selected, including 124 males and 59 females. The dialysis time before kidney transplantation was (2.9±2.1) years. Among them, there were 93 cases having cadaveric renal transplantation and 90 cases having living relative renal transplantation. The conditions of ophthalmic complications in all the patients after kidney transplantation were investigated and the incidence rate on ophthalmic complications having different dialysis time before kidney transplantation was given Chi-square test and Chi-square linear trend test. RESULTS: Among 183 patients with kidney transplantation, 95 patients (51.9%) had at least one ophthalmic complication and the rest 88 patients (48.1%) had no significant abnormality at the eye region. The most common ophthalmic complications were pinguecula/conjunctival degeneration (31 cases), the following was caligo lentis (24 cases). The main manifestations were grayish white granule and plaque turbidity occurred in posterior capsule at the posterior pole of crystaline lens. The angulus iridocornealis of 5 patients (5.3%) with cataract and glaucoma were all open-angle through the detection by gonioscope. Through visual field examination, there were 2 patients with paracentral scotoma, 2 patients with arcuate scotoma and one case with nasal step. CONCLUSION: The experiments verify that the incidence of glaucomawas relates to the dialysis time before kidney transplantation, and the incidence rate might be higher if the dialysis time is longer.