[Double-channel expansion of the subcutaneous lower abdominal tunnel for sequential inguinal lymph node dissection in penile cancer - Report of six cases].

OBJECTIVE: To investigate the safety and efficacy of the two-channel dilatation procedure for subcutaneous tunneling in the lower abdomen during pelvic lymph node dissection for penile cancer. METHODS: A retrospective analysis was conducted on the clinical data of 6 patients treated from January 2020 to December 2022 using the dual-channel expansion technique for penile cancer lymph node dissection. RESULTS: All 6 cases ( 12 sides) successfully underwent prophylactic inguinal lymph node dissection. The average laparoscopic dissection time was ( 82.50 ± 12.08) minutes per side, with an average blood loss of （28.33 ± 10.95） ml. The number of lymph nodes dissected was （11.16 ± 1.02） for the superficial group and ( 0.67 ± 0.74 ) for the deep group. Postoperative pathology was negative in all cases. The average postoperative hospital stay was （7.33 ± 1.60 ） days, with a catheter removal time of （12.00 ± 2.06）days. Postoperative complications included abnormal skin sensations in 5 sides, lower limb edema in 3 sides, lymphedema in 3 sides, and cellulitis in 1 side. During a follow-up period of （20.60 ± 12.51）months, there were no instances of tumor recurrence or metastasis in the inguinal region among the patients. CONCLUSION: The dual-channel expansion technique for inguinal lymph node dissection via a subcutaneous tunnel is a safe and feasible treatment for penile cancer. It has a low complication rate, allows for thorough dissection of inguinal lymph nodes, and offers advantages in terms of surgical time.

Co-editing PINK1 and DJ-1 Genes Via Adeno-Associated Virus-Delivered CRISPR/Cas9 System in Adult Monkey Brain Elicits Classical Parkinsonian Phenotype.

Whether direct manipulation of Parkinson's disease (PD) risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue. Here, we used an adeno-associated virus serotype 9 (AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras (SNs) of two monkey groups: an old group and a middle-aged group. After the operation, the old group exhibited all the classic PD symptoms, including bradykinesia, tremor, and postural instability, accompanied by key pathological hallmarks of PD, such as severe nigral dopaminergic neuron loss (>64%) and evident α-synuclein pathology in the gene-edited SN. In contrast, the phenotype of their middle-aged counterparts, which also showed clear PD symptoms and pathological hallmarks, were less severe. In addition to the higher final total PD scores and more severe pathological changes, the old group were also more susceptible to gene editing by showing a faster process of PD progression. These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys. Taken together, this system can effectively develop a large number of genetically-edited PD monkeys in a short time (6-10 months), and thus provides a practical transgenic monkey model for future PD studies.

Clinical efficacy of counterclockwise rotating the functional occlusal plane using micro-implant anchorage.

To evaluate the therapeutic effect of using micro-implant anchorage (MIA) to rotate the functional occlusal plane (FOP) counterclockwise. Forty skeletal class Ⅱ high-angle patients who had completed orthodontic treatment were enrolled, including 20 patients treated with MIA orthodontic system (MIA group) and the other 20 patients treated with traditional sliding straight wire appliance (control group). Cephalometric measurements on the lateral cranial radiographs before and after treatment were performed, all acquired data were statistically analyzed with SPSS 26.0. At the end of treatment, MIA group obtained better effect of FOP and mandibular plane counter-clockwise rotation than the control group. In the MIA group, the average change of FOP-frankfort horizontal plane (FH), FOP-SN and mandibular plane angle (MP-FH) angle was -4.5(-7.3, -3.7)°, (3.3)° and -1.7(-3.0, -0.9)°, respectively. In the control group, the average change of FOP-FH, FOP-SN and MP-FH angle was -0.1(-4.1, 3.0)°, (-0.1±5.1)° and -0.4(-2.4, 0.7)°, respectively. There was significant difference between the change of the two groups (all <0.05). Compared with the traditional sliding straight wire appliance, counterclockwise rotation of FOP can be more effectively reversed by using MIA orthodontic system, and the MP-FH can be reduced as well.

Novel Controlled Release Microspheric Soil Conditioner Based on the Temperature and pH Dual-Stimuli Response.

A novel type of temperature and pH dual-stimuli-responsive microspheric soil conditioner was prepared for the controlled release of urea. First, poly(N-isopropylacrylamide-co-methacrylic acid) [P(NIPAM-co-MAA)] was synthesized, and the microspheric soil conditioner was prepared on the basis of chitosan-coated P(NIPAM-co-MAA) via the emulsion cross-linking method. The structure and morphology of the microsphere were characterized by Fourier transform infrared spectroscopy, hydrogen nuclear magnetic resonance, polarization optical microscopy, and scanning electron microscopy. The microsphere showed controlled release behavior in different temperature and pH conditions, indicating good stimuli responsiveness. The plant experiment revealed that the microsphere can effectively promote plant growth in acidified soil and high-temperature conditions, and the pH value of acidified soil could be improved. In addition, the microsphere possessed good biodegradation property in the soil. Therefore, the multi-responsive microspheric soil conditioner owns a great potential value to amend soil conditions and promote plant growth in agriculture applications.

Accurate MRSA identification through dual-functional aptamer and CRISPR-Cas12a assisted rolling circle amplification.

Infectious diseases have become one of the most threatening global challenge with high morbidity and mortality, bringing great difficulties to clinical diagnosis and treatment. New strategy for high-specific and sensitive bacteria detection are urgently needed in facing the crisis of worldwide antibiotic resistance. Herein, a novel method through the integration of dual aptamer technology and CRISPR-Cas12a assisted rolling circle amplification (RCA) was present to obtain both accurate identification and high-sensitive detection of Methicillin-Resistant Staphylococcus Aureus (MRSA). The specificity inherited from the dual functionalized aptamers initiated bioconjugation to specifically recognize the protein targets on the surface of bacteria. Besides the target activity, the functionalized aptamer could also convert the protein recognition to nucleic acids signals. Through the integration of attached RCA and CRISPR-Cas12a assisted trans-cleavage, dual amplification of the nucleic acid signal was obtained. Based on this, we have extended the application of CRISPR-Cas12a from the nucleic acid detection to bacteria detection. As a result, the proposed method was demonstrated to be with significantly improved sensitivity towards MRSA detection. We believe that the novel integrated strategy would diversify the existing pool of bacterial detection and inspire the development of promising drug candidates in the future.

miR-451a is downregulated and targets PSMB8 in prostate cancer.

Abnormal expression of microRNAs (miRNAs) is frequently occurred in prostate cancer (PCa). This study was aimed to investigate the biological roles of miR-451a in PCa. Quantitative real-time PCR (qRT-PCR) and Western blot were employed to investigate the expression levels of miR-451a and proteasome (prosome, macropain) subunit, beta type, 8 (PSMB8) in PCa cell lines. Luciferase activity reporter assay was used to verify the connection between miR-451a and PSMB8. in vitro functional experiments were performed to measure the effects of miR-451a or PSMB8 on PCa cell proliferation, colony formation ability, cell invasion, and cell apoptosis. miR-451a expression was downregulated, whereas PSMB8 expression was upregulated in PCa cell lines. Luciferase activity reporter assay confirmed the direct connection between miR-451a and PSMB8. Overexpression of miR-451a inhibits PCa cell proliferation, colony formation, cell invasion and promotes cell apoptosis, while the overexpression of PSMB8 caused the opposite effects. Moreover, rescue experiments confirmed PSMB8 was a functional target of miR-451a. In conclusion, this study provides novel insights into the role of miR-451a in PCa, and the results demonstrated miR-451a could inhibit PCa progression by targeting PSMB8.

Expression of miR-182 and Foxo3a in patients with bladder cancer correlate with prognosis.

OBJECTIVE: FoxO3a is a specific tumor suppressor gene in the forkhead transcription factor O subfamily (FoxO). Studies show that its expression plays a role in bladder cancer. The abnormal expression of miR-182 in bladder cancer suggests that miR-182 may be an oncogene in bladder cancer. Bioinformatic analysis showed that there is a target complementary binding site between miR-182 and Foxo3a. In this study, the expression of miR-182 and Foxo3a in cancer tissues of patients with bladder cancer was detected. The expression of miR-182 and Foxo3a in bladder cancer tissues and their relationship with the prognosis of the patients were analyzed, and the role of miR-182 in regulating the expression of Foxo3a and the biologic process of cell proliferation and apoptosis in bladder cancer cells was explored. METHODS: Tumor tissues of patients with bladder cancer were collected and the normal bladder mucosa was used as a control. The expression of Foxo3a was detected by western blot. The expression of miR-182 and Foxo3a mRNA was detected by qRT-PCR. The relationship between miR-182, Foxo3a mRNA and the clinical features of patients was analyzed. The median expression of miR-182 and Foxo3a mRNA was bounded, and Log Rank test was used to compare the survival rate of low and high expression of miR-182 and Foxo3a mRNA. The double luciferase reporter gene assay was used to confirm a target regulatory effect between miR-182 and Foxo3a. In vitro, RT112 and T24 cells were divided into 2 groups: group miR-NC, and group miR-182 inhibitor. qRT-PCR and western blot were used to detect the expression of Foxo3a, flow cytometry was used to detect cell apoptosis, and EdU staining was used to detect cell proliferation. RESULTS: Compared with normal bladder tissue, the expression of miR-182 in bladder cancer tissue was significantly increased, and it was related to tumor size, TNM stage, and lymph node metastasis (P < 0.05). The expression of Foxo3a mRNA was significantly decreased, and was related to tumor size, TNM stage, histopathologic classification, and lymph node metastasis (P < 0.05). There was a significant negative correlation between the expression of miR-182 and Foxo3a mRNA in bladder cancer (r = -0.602, P < 0.05). The prognosis of patients with high expression of miR-182 was significantly worse than that of those with low miR-182 expression. The prognosis of patients with low expression of Foxo3a was significantly better than those with high Foxo3a. Double luciferase reporter gene experiments confirmed that there was a target regulatory relationship between miR-182 and Foxo3a. Transfection of miR-182 inhibitor significantly increased the expression of Foxo3a in RT112 and T24 cells, significantly reducing cell proliferation, and significantly increasing apoptosis. CONCLUSION: The expression of miR-182 was increased and the expression of Foxo3a was decreased in bladder cancer, which is related to prognosis. Downregulation of the expression of miR-182 can increase the expression of Foxo3a, inhibiting the proliferation of bladder cancer cells and inducing apoptosis.

Pathologically examining a minimum of three lymph nodes could better determine node negativity in patients with non-metastatic chromophobe renal cell carcinoma.

OBJECTIVE: To evaluate the prognostic value of lymph node (LN) involvement for patients with chromophobe renal cell carcinoma (chRCC) and ascertain the minimum number of LNs that need to be pathologically examined to reliably diagnose a patient with node negative chRCC. METHODS: From 2004 to 2014, non-metastatic chRCC patients receiving radical nephrectomy together with lymphadenectomy were identified from the Surveillance, Epidemiology and End Results (SEER) database. The primary outcome was overall survival (OS). RESULTS: Two hundred and forty-six patients received lymph node dissection during the surgery. Of the patients, 24 (10%) had pathologically confirmed positive LN. Multivariate Cox regression model showed that positive LN was an independent unfavorable predictor for OS (HR = 2.83, 95%CI = 1.14-6.98, P = 0.024). More importantly, LN(-) patients with at least three LNs dissected had significantly better OS compared with when 1-2 LNs were examined (P = 0.048). Multivariate analysis confirmed that in LN(-) patients, the examination of three or more LNs could independently predict better OS compared with patients with only 1-2 LNs dissected (HR≥3LNs = 0.362, 95% CI = 0.135-0.972, P = 0.044). Additionally, the likelihood of finding at least one positive LN was significantly higher on dissection of ≥3 LNs compared with examination of 1-2 LNs (15% vs 5%, P = 0.018). Decision curve analysis found a better clinical validity of the '3 LNs examined'-based classification compared with the traditional LN(-)/LN(+) classification. CONCLUSION: The proportion of positive LNs in chRCC was far from neglectable and LN metastasis could independently predict unfavorable OS. We recommended a minimum of three LNs should be pathologically examined in order to reliably determine node negative.

Alterations of Gray Matter Volume and White Matter Integrity in Maternal Deprivation Monkeys.

Maternal deprivation (MD) in rhesus monkeys has been demonstrated to be an effective model to mimic early adversity in humans because of the close phylogenetic similarity affinity. Although behavioral and hormonal abnormalities have been observed in MD monkeys, the neurobiological underpinning of the long-term deleterious effect of MD on monkeys is still unclear. In this study, we assessed emotional changes and socio-behavioral abnormalities induced by long-term MD and assessed structural alterations of gray matter volume (GMV) and white matter integrity (WMI) in 15 MD rhesus monkeys and in 15 age-, gender-matched normal controls (NC) using voxel-based morphology and voxel-based analysis methods. We found increased stereotypical behavioral durations and decreased social grooming durations in MD monkeys. Reduced GMV in the primary visual cortex (V1) and increased fractional anisotropy (FA) in the left posterior superior temporal sulcus (pSTS) was also found in MD monkeys. Moreover, the mean FA values in pSTS showed positive correlation with the stereotypical behavioral durations in MD monkeys and negative correlation with social grooming durations in NC monkeys. Our findings indicated that the deleterious effects of MD on rhesus monkeys resulted in structural abnormalities in the visual cortex and premature myelination in the pSTS. These findings provide new insights into understanding the impact of maternal deprivation on the neurological basis of brain development.

Neurons Differentiated from Transplanted Stem Cells Respond Functionally to Acoustic Stimuli in the Awake Monkey Brain.

Here, we examine whether neurons differentiated from transplanted stem cells can integrate into the host neural network and function in awake animals, a goal of transplanted stem cell therapy in the brain. We have developed a technique in which a small "hole" is created in the inferior colliculus (IC) of rhesus monkeys, then stem cells are transplanted in situ to allow for investigation of their integration into the auditory neural network. We found that some transplanted cells differentiated into mature neurons and formed synaptic input/output connections with the host neurons. In addition, c-Fos expression increased significantly in the cells after acoustic stimulation, and multichannel recordings indicated IC specific tuning activities in response to auditory stimulation. These results suggest that the transplanted cells have the potential to functionally integrate into the host neural network.

The temporary and accumulated effects of transcranial direct current stimulation for the treatment of advanced Parkinson's disease monkeys.

Transcranial direct current stimulation (tDCS) is a useful noninvasive technique of cortical brain stimulation for the treatment of neurological disorders. Clinical research has demonstrated tDCS with anodal stimulation of primary motor cortex (M1) in Parkinson's disease (PD) patients significantly improved their motor function. However, few studies have been focused on the optimization of parameters which contributed significantly to the treatment effects of tDCS and exploration of the underline neuronal mechanisms. Here, we used different stimulation parameters of anodal tDCS on M1 for the treatment of aged advanced PD monkeys induced with 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) administration, and then analyzed the temporary and accumulated effects of tDCS treatment. The results indicated anodal tDCS on M1 very significantly improved motor ability temporarily; importantly, the treatment effects of anodal tDCS on M1 were quantitatively correlated to the accumulated stimulation instead of the stimuli intensity or duration respectively. In addition, c-fos staining showed tDCS treatment effects activated the neurons both in M1 and substantia nigra (SN). Therefore, we propose that long time and continue anodal tDCS on M1 is a better strategy to improve the motor symptoms of PD than individual manipulation of stimuli intensity or duration.

1-Methyl-4-phenylpyridinium stereotactic infusion completely and specifically ablated the nigrostriatal dopaminergic pathway in rhesus macaque.

INTRODUCTION: Complete and specific ablation of a single dopaminergic (DA) pathway is a critical step to distinguish the roles of DA pathways in vivo. However, this kind of technique has not been reported in non-human primates. This study aimed to establish a lesioning method with a complete and specific ablation. METHOD: A carefully designed infusion route based on a MRI stereotactic technique was developed to deliver the highly selective dopaminergic toxin 1-methyl-4-phenylpyridinium (MPP+) unilaterally into multiple sites of compact part of substantia nigra (SNc) and striatum in monkeys. The nigrostriatal DA pathway was selected because lesioning of this pathway may induce symptoms that are suitable for evaluation. The pathological, behavioral, neuropharmacological, and clinical laboratorial data were collected to evaluate the lesioning effects. RESULT: Pathological examination revealed a complete ablation of tyrosine hydroxylase positive (TH+) neurons in the SNc, while preserving intact TH+ neurons in the ventral tegmental area (VTA) nearby. TH+ projections in the striatum were also unilaterally lost. The monkeys displayed stable (>28 weeks) rotations and symptoms which were expected with loss of DA neurons in the SNc, with rest tremor being an exception. No item implied the presence of a severe side effect caused by the operation or the intracerebral MPP+ infusion. The results suggested that rest tremor may not directly rely on the nigrostriatal pathway. CONCLUSION: Taken together, in addition to providing a specific nigrostriatal DA lesioned model, this method, combined with brain stimulation or other techniques, can be applied as a powerful tool for the complete lesion of any desired DA pathway in order to study its specific functions in the brain.

Cystic renal cell carcinoma: a report of 67 cases including 4 cases with concurrent renal cell carcinoma.

BACKGROUND: Cystic renal cell carcinoma (CRCC) is relatively rare; CRCC is frequently misdiagnosed as a benign renal cyst. CRCC carries an excellent prognosis following surgical treatment. The aim of our study was to summarize the management of CRCC and to characterize the prognosis of affected patients. METHODS: A retrospective study of 67 patients with CRCC was conducted at our center between January 2005 and April 2013. Patient prognosis as well as the clinical manifestations, imaging characteristics, treatment, and pathologic features of CRCC were summarized based on available medical record data. RESULTS: We identified 67 cases of CRCC, representing 2.5% of all renal cell carcinoma cases. The tumor was discovered incidentally in 70% of the cases. Ultrasonography was found to be a useful screening tool, but computed tomography remains the imaging study of choice for identifying malignant features. Magnetic resonance imaging can be used in equivocal cases. Regarding treatment, radical nephrectomy was performed in 52% of the cases, and partial nephrectomy was selected in the remaining 48% of cases. None of the 46 patients (68% of the study group) available for follow-up showed any evidence of recurrence. CONCLUSIONS: CRCC is an uncommon subtype of renal cell carcinoma, occurring in 2.5% of cases. CRCC carries an excellent prognosis after surgical treatment. Partial nephrectomy should be regarded as the preferred surgical technique for CRCC.

Perforator pedicled propeller flaps for soft tissue coverage of lower leg and foot defects.

OBJECTIVE: To investigate the efficiency of perforator pedicled propeller flaps for soft tissue coverage of lower leg and foot defects. METHODS: Twenty patients (12 male, 8 females; mean age 28 years, range, 5-75) with soft tissue defects of the lower leg and foot were retrospectively reviewed. Their defects had been repaired with perforator pedicled propeller flaps from September 2011 to October 2013 and included five cases of injuries caused by spokes, four of infection with postoperative skin necrosis, two of dorsal skin defects caused by heavy objects and nine caused by car accidents. The areas of soft tissue defect were from 2 cm × 8 cm to 10 cm × 20 cm. Fifteen cases had terminal branch of the peroneal artery perforator flaps and five posterior tibia artery perforator flaps, flap size ranging from 5 cm × 11 cm to 12 cm × 28 cm. Color Doppler ultrasound was used to locate all perforator vessels, the calibers of which ranged from 0.8 mm to 1.0 mm. RESULTS: The intraoperative coincidence rate of the color Doppler ultrasound was 96.7%. The donor sites were sutured directly in 12 cases and skin grafted in 8. One case had a venous crisis within 24 h that was treated by removal some sutures and drainage. All cases were followed up for 1-18 months; all flaps survived well and pedicles had a satisfactory appearance. The patients were extremely satisfied with the results for repair. CONCLUSION: Perforator pedicled propeller flaps have the advantages over other pedicle flap of being simple, safe, and effective and not involving vascular anastomosis.

A new method for piercing the tentorium cerebelli for implanting fragile electrodes into the brain stem in the rhesus monkey (Macaca mulatta).

Recent developments in neuron recording techniques include the invention of some fragile electrodes. The fragility of these electrodes impedes their successful use in deep brain recordings because it is difficult to penetrate the electrodes through the dura mater, especially the tentorium cerebelli (TC) enclosing the cerebellum and brain stem. This paper reports a new method to pierce the TC for inserting fragile electrodes into the inferior colliculus of rhesus monkeys. Briefly, a unique tool kit, consisting of needles with sharp tips, a guide tube and an "impactor," was used in a multistep protocol to pierce the TC. The impactor provided a brief force that quickly thrusts the needles through the meninges without causing significant damage to the brain tissue under the TC. Using this novel approach, tetrodes were successfully implanted into the inferior colliculus of a rhesus monkey and neuronal discharge signals were recorded. This method, which is simple, convenient and economical, allows neurophysiologists to study the electrophysiological characteristics of deep brain structures under the TC with advanced, albeit fragile, electrodes.

Blockage of glucocorticoid receptors during memory acquisition, retrieval and reconsolidation prevents the expression of morphine-induced conditioned place preferences in mice.

Association between the reward caused by consuming drugs and the context in which they are consumed is essential in the formation of morphine-induced conditioned place preference (CPP). Glucocorticoid receptor (GRs) activation in different regions of the brain affects reward-based reinforcement and memory processing. A wide array of studies have demonstrated that blockage of GRs in some brain areas can have an effect on reward-related memory; however, to date there have been no systematic studies about the involvement of glucocorticoids (GCs) in morphine-related reward memory. Here, we used the GR antagonist RU38486 to investigate how GRs blockage affects the sensitization and CPP behavior during different phases of reward memory included acquisition, retrieval and reconsolidation. Interestingly, our results showed RU38486 has the ability to impair the acquisition, retrieval and reconsolidation of reward-based memory in CPP and sensitization behavior. But RU38486 by itself cannot induce CPP or conditioned place aversion (CPA) behavior. Our data provide a much more complete picture of the potential effects that glucocorticoids have on the reward memory of different phases and inhibit the sensitization behavior.

Lesion of olfactory epithelium attenuates expression of morphine-induced behavioral sensitization and reinstatement of drug-primed conditioned place preference in mice.

Previous studies have shown that olfactory impairment by disrupting the olfactory epithelium prior to morphine administration attenuated the development addiction-related behaviors. However, it is unclear whether olfactory impairment will affect the expression of already established addiction-related behaviors. To address this issue, mice were conditioned with morphine to induce behavioral sensitization and condition placed preference (CPP). After an abstinence period, the animals were subjected to either an intranasal ZnSO(4) effusion (ZnE) or sham treatment with saline. Behavioral sensitization and CPP reinstatement were evaluated 24h later, as well as the expression of c-Fos protein, a marker of activated neural sites, in brain regions of interest. It was found that ZnE treatment attenuated morphine-induced behavioral sensitization and reinstatement of CPP. Compared to the saline-treated ones, the ZnE-treated animals showed reduced c-Fos expression in the nucleus accumbens (NAc) associated with behavioral sensitization, and in the NAc, cingulate cortex, dentate gyrus, amygdala, lateral hypothalamus and ventral tegmental area associated with CPP reinstatement. Together, these results demonstrated that acute olfactory impairment could attenuate already established addiction-related behaviors and expression of c-Fos in drug addiction related brain regions, perhaps by affecting the coordination between reward and motivational systems in the brain.

Combined action of MK-801 and ceftriaxone impairs the acquisition and reinstatement of morphine-induced conditioned place preference, and delays morphine extinction in rats.

OBJECTIVE: It is well established that glutamate and its receptors, particularly the N-methyl-D-aspartate receptor (NMDAR), play a significant role in addiction and that the inhibition of glutamatergic hyperfunction reduces addictive behaviors in experimental animals. Specifically, NMDAR antagonists such as MK-801, and an inducer of the expression of glutamate transporter subtype-1 (GLT-1) (ceftriaxone) are known to inhibit addictive behavior. The purpose of this study was to determine whether the combined action of a low dose of MK-801 and a low dose of ceftriaxone provides better inhibition of the acquisition, extinction, and reinstatement of morphine-induced conditioned place preference (CPP) than either compound alone. METHODS: A morphine-paired CPP experiment was used to study the effects of low doses of MK-801, ceftriaxone and a combination of both on reward-related memory (acquisition, extinction, and reinstatement of morphine preference) in rats. RESULTS: A low dose of neither MK-801 (0.05 mg/kg, i.p.) nor ceftriaxone (25 mg/kg, i.p.) alone effectively impaired CPP behaviors. However, when applied in combination, they reduced the acquisition of morphine-induced CPP and completely prevented morphine reinstatement. Their combination also notably impaired the extinction of morphine-induced CPP. CONCLUSION: The combined action of a low dose of an NMDAR antagonist (MK-801) and GLT-1 activation by ceftriaxone effectively changed different phases of CPP behavior.