DNMT3a contributes to the development and maintenance of bone cancer pain by silencing Kv1.2 expression in spinal cord dorsal horn.

Abstract: Metastatic bone tumor-induced changes in gene transcription and translation in pain-related regions of the nervous system may participate in the development and maintenance of bone cancer pain. Epigenetic modifications including DNA methylation regulate gene transcription. Here, we report that intrathecal injection of decitabine, a DNA methyltransferase (DNMT) inhibitor, dose dependently attenuated the development and maintenance of bone cancer pain induced by injecting prostate cancer cells into the tibia. The level of the de novo DNMT3a, but not DNMT3b, time dependently increased in the ipsilateral L4/5 dorsal horn (not L4/5 dorsal root ganglion) after prostate cancer cells injection. Blocking this increase through microinjection of recombinant adeno-associated virus 5 (AAV5) expressing Dnmt3a shRNA into dorsal horn rescued prostate cancer cells-induced downregulation of dorsal horn Kv1.2 expression and impaired prostate cancer cells-induced pain hypersensitivity. In turn, mimicking this increase through microinjection of AAV5 expressing full-length Dnmt3a into dorsal horn reduced dorsal horn Kv1.2 expression and produced pain hypersensitivity in the absence of prostate cancer cells injection. Administration of neither decitabine nor virus affected locomotor function and acute responses to mechanical, thermal, or cold stimuli. Given that Dnmt3a mRNA is co-expressed with Kcna2 mRNA (encoding Kv1.2) in individual dorsal horn neurons, our findings suggest that increased dorsal horn DNMT3a contributes to bone cancer pain through silencing dorsal horn Kv1.2 expression. DNMT3a may represent a potential new target for cancer pain management.

Impaired neuropathic pain and preserved acute pain in rats overexpressing voltage-gated potassium channel subunit Kv1.2 in primary afferent neurons.

Voltage-gated potassium (Kv) channels are critical in controlling neuronal excitability and are involved in the induction of neuropathic pain. Therefore, Kv channels might be potential targets for prevention and/or treatment of this disorder. We reported here that a majority of dorsal root ganglion (DRG) neurons were positive for Kv channel alpha subunit Kv1.2. Most of them were large and medium, although there was a variety of sizes. Peripheral nerve injury caused by lumbar (L)5 spinal nerve ligation (SNL) produced a time-dependent reduction in the number of Kv1.2-positive neurons in the ipsilateral L5 DRG, but not in the contralateral L5 DRG. Such reduction was also observed in the ipsilateral L5 DRG on day 7 after sciatic nerve axotomy. Rescuing nerve injury-induced reduction of Kv1.2 in the injured L5 DRG attenuated the development and maintenance of SNL-induced pain hypersensitivity without affecting acute pain and locomotor function. This effect might be attributed to the prevention of SNL-induced upregulation of endogenous Kv1.2 antisense RNA, in addition to the increase in Kv1.2 protein expression, in the injured DRG. Our findings suggest that Kv1.2 may be a novel potential target for preventing and/or treating neuropathic pain.

[Spermicidal effect of alcohol extracts from different ratios of Sophora flavescens Ait/Chinese Bulbul in vitro].

OBJECTIVE: To observe the spermicidal effect of alcohol extracts from different ratios of Sophora flavescens Ait/Chinese Bulbul in vitro. METHODS: Semen samples aseptically obtained by masturbation and prepared by density gradient centrifugation from 15 healthy men were incubated in the alcohol extracts from 9 different ratios of Sophora flavescens Ait/Chinese Bulbul for 20 seconds, 2 minutes and 4 minutes. Then the motility and movement parameters of the sperm were detected by computer-assisted semen analysis, and the minimal effective concentrations of the instant spermicidal effect of the extracts were determined. RESULTS: At the ratio of 3:1, the extract at 0.5 mg/ml significantly inhibited the sperm motility and other sperm movement parameters VCL, VSL, VAP, ALH, WOB and MAD, as compared with the control group. The minimal effective concentration of the instant spermicidal effect of the extracts was 3.5 mg/ml at 3:1. CONCLUSION: The alcohol extracts from Sophora flavescens Ait and Chinese Bulbul at the ratio of 3:1 have the best spermicidal effect in vitro.

Successful Anesthesia Management in a Patient With Type A Aortic Dissection Complicated by Renal Failure and Suspected Coronavirus Disease.

Stanford type A acute aortic dissection (AAD) is a life-threatening illness that presents with chest pain and hemodynamic instability. AAD prompt and accurate evaluation and management are critical for survival as it is a cardiac surgical emergency. The initial treatment of AAD mandates strict blood pressure stabilization with intravenous antihypertensive medications. The progressive nature of the disease will increase the mortality as time elapses between diagnosis and surgical intervention. In addition, the patient's blood pressure control is challenged in the presence of renal failure requiring hemodialysis. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 or named 2019-nCoV) pneumonia was a newly underrecognized illness (COVID-19 [coronavirus disease 2019]). COVID-19 can cause severe acute respiratory distress syndrome, acute kidney injury, heart injury, and liver dysfunction, which would aggravate the progress of aortic dissection. In this article, we report the successful anesthesia management in a pneumonia patient with AAD complicated with renal failure during the COVID-19 epidemic period, who underwent emergency surgery and deep hypothermic circulatory arrest repair.

Blood Flow Index and Skin Temperature Measured by Laser Speckle Contrast Imaging and Infrared Thermography After Specific Ultrasound-Guided Blocking of the C6, C7 Nerve Root: A Case Report.

Ultrasound-guided cervical nerve root block is the technique of injecting local anesthetic or steroid into the intertubercular groove outside of the cervical foramen under real-time ultrasound guidance. It has established diagnostic and therapeutic employment for patients suffering symptoms caused by cervical disc disorders and foraminal stenosis. Measures of selective nerve root blockade would have vital clinical utility, especially in multilevel disease or combined with atypical symptoms, where subjective assessment is difficult. This case reported a 47-year-old male patient who suffered from severe radicular pain confined to the index and middle fingers, with weakness of the right upper extremity. He was successfully treated with selective C6, C7 nerve root under ultrasound guidance. For confirmation of the proper analgesia range, laser speckle contrast imaging (LSCI) and infrared thermography (IT) were both constructed over the dorsum of the hands and forearms during the procedure. The results of the off-line analysis suggested LSCI and IT might be considered as alternative measurements for the selective cervical nerve root.

Study of the disparity of noninvasive and invasive blood pressure measured by the Philips Intellivue MP50 monitor in surgeries inducing gall cardiac reflex under general anesthesia.

OBJECTIVE: The aim of the present study was to comprehensively compare noninvasive and invasive blood pressure (BP) measured by the Philips Intellivue MP50 monitor in surgeries that may induce gall cardiac reflex under general anesthesia. PARTICIPANTS AND METHODS: Seventy-eight patients undergoing cholecyst or bile duct operations under general anesthesia were enrolled in our study. Both invasive (intraradial, femoral, or dorsalis pedis artery) and noninvasive (oscillometric) BP were monitored by the Philips Intellivue MP50 monitor simultaneously. Data were analyzed using Bland-Altman plots. RESULTS: In a supine position during operations, between intraradial and oscillometric measurements, the bias and precision (mmHg) were 9.34±12.98 and 12.47±10.00 for systolic blood pressure (SBP) and 3.26±8.22 and 6.53±5.97 for diastolic blood pressure (DBP), between intrafemoral and oscillometric measurements, the bias and precision (mmHg) were 14.40±14.38 and 16.93±11.28 for SBP and 4.35±9.72 and 7.52±7.54 for DBP, between intradorsalis pedis and oscillometric measurements, the bias and precision (mmHg) were 15.69±14.37 and 16.91±12.91 for SBP and 0.99±7.69 and 5.67±5.27 for DBP. CONCLUSION: The oscillometric BP showed poor agreement with intra-arterial BP in cholecyst or bile duct surgeries that may induce gall cardiac reflex under general anesthesia. Therefore, according to the present data, application of oscillometric BP measured by the Philips Intellivue MP50 monitor in these surgery patients under general anesthesia cannot be recommended generally.

Electroejaculation combined with assisted reproductive technology in psychogenic anejaculation patients refractory to penile vibratory stimulation.

BACKGROUND: Psychogenic anejaculation (PAE) is a rare disease in clinic seriously affecting quality of life, especially fertility of patients. However, there is not an effective treatment for these patients. The aim of the study was to evaluate the results of treatment with electroejaculation (EEJ), intrauterine insemination (IUI), and intracytoplasmic sperm injection (ICSI) in patients with PAE who were refractory to penile vibratory stimulation (PVS). METHODS: Twenty PVS-refractory patients with PAE were included in the retrospective study. Semen was retrieved through EEJ procedures. According to the semen quality and patient preference, IUI or ICSI was selected. RESULTS: A total of 23 procedures of EEJ were performed in 20 patients. Sperm was successfully retrieved in all of the cases. Yielded semen was used in a total of 16 ICSI procedures in 6 couples. The fertilization rate was 54.0%. The ICSI cycles resulted in three pregnancies; of these one resulted in a live birth. Fourteen couples were treated with 26 IUI cycles, which resulted in live births in five pregnancies. CONCLUSIONS: EEJ offers an encouraging pregnancy opportunity for men with PAE who were refractory to PVS. The combined use of EEJ and assisted reproductive techniques is excellent management for PAE infertility. In our study, EEJ resulted in pregnancies and the birth of six healthy children.

Risk factors for acute kidney injury after orthotopic liver transplantation: A single-center data analysis.

Acute kidney injury (AKI) is a common complication following orthotopic liver transplantation (OLT) and is associated with increased morbidity and mortality. The aim of the current study was to determine the risk factors for AKI in patients undergoing OLT. A total of 103 patients who received OLT between January 2015 and May 2016 in Tongji Hospital, China, were retrospectively analyzed. Their demographic characteristics and perioperative parameters were collected, and AKI was diagnosed using 2012 Kidney Disease: Improving Global Outcomes (KDIGO) staging criteria. It was found that the incidence of AKI was 40.8% in this cohort and AKI was significantly associated with body mass index, urine volume, operation duration (especially > 480 min), and the postoperative use of vasopressors. It was concluded that relative low urine output, long operation duration, and the postoperative use of vasopressors are risk factors for AKI following OLT.

[Experimental study of augmentation phalloplasty using tunica vaginalis grafts in bilateral albuginea of penile corpus].

OBJECTIVE: To study the effect and complications of augmentation phalloplasty by implanting autogenous tunica vaginalis grafts into the bilateral albuginea of the penile corpus. METHODS: Five mongrel dogs underwent augmentation phalloplasty with bilateral autogenous tunica vaginalis grafts. The increase in the volume of the corpora cavernosa was achieved by applying autogenous tunica vaginalis grafts to longitudinal openings made bilaterally in the albuginea along the whole length of the penile cavernous corpora. The perimeter of the penile cavernous corpora was measured in the flaccid state and during erection and dynamic infusion cavernosometry (DIC) was conducted for each dog before and after the operation. Histology of the penis was examined using hematoxylin and eosin stains. RESULTS: The average increase in the perimeter of the penile cavernous corpora was 21. 1% in the erectile state but unobvious in the flaccid state 3 months after surgery. There were no statistically significant differences in cavernosometric parameters before and after the operation, and neither were there any other severe postoperative complications. The grafts had mild inflammatory reaction and fibrosis with apparent reformation of the tunica albuginea over the patch site. CONCLUSION: The augmentation phalloplasty technique with bilateral autogenous tunica vaginalis grafts was proved to be effective and reliable with few complications, particularly conspicuous in increasing the volume of the erectile tissues during erection.

Effects of ligustrazine on the contraction of isolated rabbit corpus cavernosum strips.

To investigate the role of ligustrazine on relaxation of the isolated rabbit corpus cavernosum tissue in vitro, the effects of ligustrazine on the corpus cavernosum were observed by using experimental method of smooth muscle strips. Concentration-responses to phenylephine (PE) and KCl were recorded. The results showed that ligustrazine concentration-dependently depressed the contraction response of smooth muscle strips induced by PE. The maximum percentage relaxation of cavernosal strips by ligustrazine was 74.1% +/- 6.2% (compared with control: 21.9% +/- 5.6%, P < 0.01). Ligustrazine concentration-dependently reduced the amplitude of the contraction induced by cumulative doses of PE or KCl, shifted the cumulative concentration response curves of PE and KCI to the right and depressed their maximal responses. It was concluded that ligustrazine could significantly relax the cavernosal muscle contraction induced by PE in vitro. The results suggested that ligustrazine inhibited calcium ion influx.

[Penile vibratory stimulation and electroejaculation in the treatment of ejaculatory dysfunction].

The fertility potential of infertile men can be enhanced to a great extent by the application of assisted reproduction techniques such as intrauterine insemination or in-vitro fertilization with or without intracytoplasmic sperm injection, but how to obtain semen from men with ejaculatory dysfunction remains a problem. The development and refinement of penile vibratory stimulation (PVS) and electroejaculation (EEJ) have significantly brightened the prospects for the treatment of ejaculatory dysfunction. Because vibratory stimulation is non-invasive and easy to perform, and needs no anaesthesia, it is preferred by most of the patients to EEJ, and recommended to be the first choice of treatment for ejaculatory dysfunction. Approximately 80% of all ejaculatory dysfunction men with an intact ejaculatory reflex arc (above T10 ) can obtain antegrade ejaculation by PVS. Any condition which affects the ejaculatory mechanism of the central and/or peripheral nervous system including surgical nerve injury may be treated successfully by EEJ. The purpose of this review is to present the current understanding of PVS and EEJ procedures and their clinical use in men with ejaculatory dysfunction.

[Effect of chuanxiongzine on the relaxation of isolated rabbit corpus cavernosum tissues].

OBJECTIVE: To investigate the effects of chuanxiongzine on the relaxation of isolated rabbit corpus cavernosum tissues. METHODS: Observations were made on the relaxing effect of chuanxiongzine on phenylephrine (PE)-precontracted corpus cavernosum and cavernosal strips preincubated with L-NAME, and ODQ, using endothelium removal and experimental method of smooth muscle strips. The effect of chuanxiongzine on cAMP and cGMP levels in corpus cavernosum was measured by 125I radioimmunoassay. RESULTS: Chuanxiongzine had concentration-dependent relaxing effect on the cavernosal muscle contraction induced by PE (EC50 1.58 x 10(-4) mol/L). This relaxing effect was partially antagonized by ODQ and not blocked by L-NAME or endothelium removal. In PE-precontracted cavernosal strips, chuanxiongzine induced relaxation accompanied with an increase in cAMP and cGMP levels. CONCLUSION: Chuanxiongzine was effective in relaxing cavernosal muscle precontracted by PE in vitro. And its relaxing effect may be enhanced by increasing cGMP and cAMP levels in the corpus cavernosum.

[One case report of clitoral priapism and literature review].

OBJECTIVE: To investigate the clinical characteristics of female clitoral priapism. METHODS: A 29-year-old case of painful priapism of the clitoris lasting 60 hours was reported. The etiology, diagnosis and treatment of clitoral priapism were discussed and the literature reviewed. RESULTS: The patient was cured by intracavernous administration of adrenaline. CONCLUSION: Clitoral priapism is extraordinarily rare. It is primarily due to oral antidepressant drugs or pelvic malignant neoplasm which leads to tissue infiltration of the clitoral veins and obstruction of the clitoral corporeal blood flow. The efficient therapeutic approach to clitoral priapism is intracavernous administration of alpha-agonists similar to the treatment of penile priapism.

Role of ATP-sensitive potassium channels in modulating nociception in rat model of bone cancer pain.

Bone cancer pain is a major clinical problem and remains difficult to treat. ATP-sensitive potassium (KATP) channels may be involved in regulating nociceptive transmission at the spinal cord level. We determined the role of spinal KATP channels in the control of mechanical hypersensitivity in a rat model of bone cancer pain. The rat model of bone cancer pain was induced by implanting rat mammary gland carcinoma cells (Walker256) into the tibias. KATP modulators (pinacidil and glibenclamide) or the specific Kir6.2-siRNA were injected via an intrathecal catheter. The mechanical withdrawal threshold of rats was tested using von Frey filaments. The Kir6.2 mRNA and protein levels were measured by quantitative PCR and western blots, respectively. Intrathecal injection of pinacidil, a KATP channel opener, significantly increased the tactile withdrawal threshold of cancer cell-injected rats in a dose-dependent manner. In contrast, intrathecal delivery of glibenclamide, a KATP channel blocker, or the specific Kir6.2-siRNA significantly reduced the tactile withdrawal threshold of cancer cell-injected rats. The mRNA and protein levels of Kir6.2 in the spinal cord of cancer cell-injected rats were significantly lower than those in control rats. Our findings suggest that the KATP channel expression level in the spinal cord is reduced in bone cancer pain. Activation of KATP channels at the spinal level reduces pain hypersensitivity associated with bone cancer pain.

Protein kinase B/Akt is required for complete Freund's adjuvant-induced upregulation of Nav1.7 and Nav1.8 in primary sensory neurons.

UNLABELLED: Voltage-gated sodium channels (Nav) are essential for the generation and conduction of action potentials. Peripheral inflammation increases the expression of Nav1.7 and Nav1.8 in dorsal root ganglion (DRG) neurons, suggesting that they participate in the induction and maintenance of chronic inflammatory pain. However, how Nav1.7 and Nav1.8 are regulated in the DRG under inflammatory pain conditions remains unclear. Using a complete Freund's adjuvant (CFA)-induced chronic inflammatory pain model and Western blot analysis, we found that phosphorylated Akt (p-Akt) was significantly increased in the ipsilateral L4/5 DRGs of rats on days 3 and 7 after intraplantar CFA injection. Immunohistochemistry showed that the percentage of p-Akt-positive neurons in the DRG was also significantly increased in the ipsilateral L4/5 DRGs at these time points. Moreover, CFA injection increased the colocalization of p-Akt with Nav1.7 and Nav1.8 in L4/5 DRG neurons. Pretreatment of rats with an intrathecal injection of Akt inhibitor IV blocked CFA-induced thermal hyperalgesia and CFA-induced increases in Nav1.7 and Nav1.8 in the L4/5 DRGs on day 7 after CFA injection. Our findings suggest that the Akt pathway participates in inflammation-induced upregulation of Nav1.7 and Nav1.8 expression in DRG neurons. This participation might contribute to the maintenance of chronic inflammatory pain. PERSPECTIVE: This article presents that inhibition of Akt blocks CFA-induced thermal hyperalgesia and CFA-induced increases in dorsal root ganglion Nav1.7 and Nav1.8. These findings have potential implications for use of Akt inhibitors to prevent and/or treat persistent inflammatory pain.

mTOR and its downstream pathway are activated in the dorsal root ganglion and spinal cord after peripheral inflammation, but not after nerve injury.

Protein translation controlled through activation of mammalian target of rapamycin (mTOR) participates in many physiological and pathological processes. However, whether such activation is required for chronic pain is still unknown. Here, we examined activation of the mTOR signaling pathway during complete Freund's adjuvant (CFA)-induced chronic inflammatory pain and L5 spinal nerve ligation (SNL)-induced neuropathic pain in rats. Western blot analysis showed significantly increased levels of phosphorylated mTOR (p-mTOR) and phosphorylated p70S6 kinase 1 (p-S6K1, a downstream effector of mTOR) in the ipsilateral L4/5 spinal cord 2h, 1 day, 3 days, and 7 days after intraplantar CFA injection and in the ipsilateral L4/5 dorsal root ganglions (DRGs) 1 and 3 days after CFA injection. Immunohistochemistry also demonstrated increases in number of p-mTOR-labeled neurons in the ipsilateral L4/5 DRGs and in density of p-mTOR-labeled immunoreactivity in the ipsilateral L4/5 superficial dorsal horn 1 day after CFA injection. Moreover, intrathecal administration of rapamycin, a selective inhibitor of mTOR, significantly blocked CFA-induced mechanical allodynia and thermal hyperalgesia 1 day post-CFA injection. Interestingly, expression of neither p-mTOR nor p-S6K1 was markedly altered on days 3, 7, or 14 after L5 SNL in L5 spinal cord or DRG. These findings indicate that in DRG and spinal cord, mTOR and S6K1 are activated during chronic inflammatory pain, but not during neuropathic pain. Our results strongly suggest that mTOR and its downstream pathway contribute to the development of chronic inflammatory pain.

Fresh semen quality in ejaculates produced by nocturnal emission in men with idiopathic anejaculation.

OBJECTIVE: To compare the quality of fresh semen samples collected by nocturnal emission (NE), penile vibratory stimulation (PVS), or electroejaculation (EEJ) from idiopathic anejaculation patients with those collected by masturbation from healthy control subjects. DESIGN: Retrospective clinical study. SETTING: University medical center. PATIENT(S): Ninety-one men with idiopathic anejaculation. INTERVENTION(S): NE, PVS, EEJ, computer-assisted semen analysis system. MAIN OUTCOME MEASURE(S): Sperm volume, concentration, motility, and morphology. RESULT(S): The mean percentage of motile sperm in the NE group was 30.6% ± 3.5% (range 11.2%-50.9%), which is 1.5- and 1.9-fold higher than that of the PVS (20.5% ± 1.3%, range 6.9%-40.2%) and EEJ (16.1% ± 0.9%, range 3.7%-27.3%) groups, respectively. The percentage of sperm that have normal morphology in the NE group was 61.4% ± 4.8%, which is significantly higher than that in the PVS (38.4% ± 2.8%) and EEJ groups (29.5% ± 1.7%), respectively. However, compared with the healthy control group, the sperm motility and normal morphology in the NE, PVS, and EEJ groups were significantly lower. The sperm concentration in the NE group (149.4 ± 16.3 × 10(6) mL(-1)) was similar to that of the PVS group (143.8 ± 13.1 × 10(6) mL(-1)) but was higher than in the EEJ group (96.3 ± 8.6 × 10(6) mL(-1)) and lower than in the control group (225.0 ± 15.0 × 10(6) mL(-1)). Furthermore, the semen volume in the NE group (2.6 ± 0.2 mL) was lower than that of the control group (3.0 ± 0.1 mL). CONCLUSION(S): This study is the first to report on the quality of fresh semen obtained by NE in patients with idiopathic anejaculation. Sperm motility and normal morphology were higher in patients who collected sperm by NE than in those who collected sperm by PVS and EEJ.

Non-invasive monitoring of blood pressure using the Philips Intellivue MP50 monitor cannot replace invasive blood pressure techniques in surgery patients under general anesthesia.

The Philips Intellivue MP50 monitor provides a method for non-invasive, near-continuous blood pressure (BP) monitoring and is designed to be an alternative to direct intra-arterial BP (IABP) measurement. However, no studies have specifically compared non-invasive and invasive BP measurements using the monitor. The present retrospective study observed 515 patients undergoing surgery with general anesthesia, whose invasive (intra-radial, femoral or dorsalis pedis artery) and non-invasive (oscillometric) BP (NIBP) were monitored simultaneously using the monitor. These data were analyzed using correlations, regressions and Bland-Altman plots. The patients were placed in a supine position during surgery. The correlation data for invasive BP and NIBP measurements were: for intra-radial measurements, r2=0.51 (bias and precision, 11.04±15.22 and 14.76±11.64 mmHg, respectively) for systolic BP (SBP) and r2=0.27 (6.17±11.95 and 9.77±9.25 mmHg, respectively) for diastolic BP (DBP); for intra-femoral measurements: r2=0.57 (14.79±14.55 and 17.15±11.68 mmHg, respectively) for SBP and r2=0.45 (4.12±9.70 and 7.49±7.40 mmHg, respectively) for DBP; and for intra-dorsalis pedis measurements: r2=0.33 (13.00±16.81 and 17.34±12.28 mmHg, respectively) for SBP and r2=0.30 (0.17±11.27 and 8.44±7.46 mmHg, respectively) for DBP. According to this data, the NIBP measured by the Philips Intellivue MP50 monitor showed low positive correlations and poor agreement with the IABP, as calculated by Bland-Altman analysis. Therefore, the use of oscillometric BP measured by the monitor in surgery patients under general anesthesia is not generally recommended.

Isoflurane post-conditioning protects primary cultures of cortical neurons against oxygen and glucose deprivation injury via upregulation of Slit2/Robo1.

Different mechanisms have been suggested to contribute to isoflurane-mediated neuroprotection. Previous studies have suggested that the protein Slit can abrogate neuronal death in mixed neuronal-glial cultures exposed to oxygen-glucose deprivation (OGD) and reperfusion (OGD/R). We hypothesized that isoflurane increases the expression of Slit and its receptor Robo when cortical neurons are exposed to OGD/R. To test this hypothesis, we exposed primary cortical neurons to OGD for 90 min and reperfusion for 24h and investigated how isoflurane post-conditioning affected cell survival and expression of Slit2 and receptors Robo1 and Robo4. Cell survival increased after administration of isoflurane, as assessed by the lactate dehydrogenase assay, trypan blue analysis, and propidium iodide staining. Western blot analysis showed that cleaved caspase-3 was increased after OGD/R(P<0.01) but reduced by isoflurane post-conditioning. Real-time PCR and Western blot analysis showed that the expression levels of Slit2 and Robo1, but not Robo4, were increased after OGD/R (P<0.5) and increased even further by isoflurane post-conditioning (P<0.01). Our results suggest that isoflurane post-conditioning markedly attenuates apoptosis and necrosis of cortical neurons exposed to OGD/R possibly in part via elevation of Slit2 and Robo1 expression. These findings provide a novel explanation for the pleiotropic effects of isoflurane that could benefit the central nervous system.