Objective: LMJ07 is a novel cannabinoid receptorl (CB1R) selective antagonist discovered by our lab. In the present study, its affinity and antagonistic activity against CB1R were evaluated at the molecular and cellular levels by receptor binding experiment, CB1R internalization experiment and by monitoring the change in cytoskeletal and intracellular signal induced by CB1R activation. Methods: With the CB1R selective antagonist rimonabant (SR141716A) as control, the affinity and selectivity of LMJ07 to CB1R and CB2R were assayed by radioligand binding assays, and the G protein-independent antagonistic activity against cannabinoid receptor (CBR) was assayed by enhanced green fluorescein protein (EGFP)-CBR internalization with hierarchiae cluscer anclysis (HCA) analysis. At the same time, we evaluated the changes in cytoskeletal and the intracellular cAMP levels in response to LMJ07 treatment in CHO-CB1 cells by Cellkey label-free assays and homogeneous time-resolved fluorescence (HTRF). Additionally, we also confirmed the CB1R antagonistic efficacy of LMJ07 by detecting the content of Ca2+ in primary cultured hippocampal neuronal cells which could express CB1R with continuous fluorescence detection technology. Results: LMJ07 is a selective CB1R antagonist with high affinity, which can selectively antagonize receptor endocytosis induced by CB1RactivationIt’s affinity and antagonistic efficacy to CB1R were equal to those of rimonabant. In CHO-CB1 cells, LMJ07 (0.01-10μmol/L)could dose-dependently inverse the change in cytoskeletal as well as the increase in intracellular cAMP induced by CBR agonist Win55212-2. In the primary cultured hippocampal neuronal cells, LMJ07 (10 nmol/L-1 μmol/L) could block the increase in [Ca2+] induced by CB1R agonist Win55212-2. Conclusion: LMJ07 is a new selective CB1R antagonist, which shows equal affinity and antagonistic activity against CB1R as the widely accepted CB1R antagonist rimonabant. In addition, the combination of high content analysis (HCS) and Cellkey label-free assay provides a better research tool for rapid and high throughput screening of novel CB1R antagonists.

Follicle-stimulating hormone (FSH) is synthesized and secreted by the anterior pituitary, which binds to its receptors expressed on the membrane of Sertoli cells in the testis to bring about spermatogenesis. With the development of DNA sequencing technology, FSH SNPs rs10835638 and FSHR SNPs rs6165, rs6166, and rs1394205 were detected, which might directly affect the expression of FSH and activity of FSHR, resulting in male spermatogenic dysfunction. This review focuses on the relationship of FSH and FSHR gene polymorphisms with male infertility.
Follicle Stimulating Hormone ; genetics ; Humans ; Infertility, Male ; genetics ; Male ; Polymorphism, Single Nucleotide ; Receptors, FSH ; genetics ; Sertoli Cells ; Spermatogenesis ; Testis

The main treatment strategies for chronic pancreatitis in young patients include therapeutic endoscopic retrograde cholangio-pancreatography (ERCP) intervention and surgical intervention. Therapeutic ERCP intervention is performed much more extensively for its minimally invasive nature, but a part of patients are referred to surgery at last. Historical and follow-up data of 21 young patients with chronic pancreatitis undergoing duodenum-preserving total pancreatic head resection were analyzed to evaluate the outcomes of therapeutic ERCP intervention and surgical intervention in this study. The surgical complications of repeated therapeutic ERCP intervention and surgical intervention were 38% and 19% respectively. During the first therapeutic ERCP intervention to surgical intervention, 2 patients developed diabetes, 5 patients developed steatorrhea, and 5 patients developed pancreatic type B pain. During the follow-up of surgical intervention, 1 new case of diabetes occurred, 1 case of steatorrhea recovered, and 4 cases of pancreatic type B pain were completely relieved. In a part of young patients with chronic pancreatitis, surgical intervention was more effective than therapeutic ERCP intervention on delaying the progression of the disease and relieving the symptoms.

Cannabinoid 1 receptor (CB1R) is one of most important targets for the treatment of obesity. However, the clinical application of CB1R antagonist rimonabant is restricted because of the central nervous system-related unwanted liabilities. Peripherally restricted CB1R antagonist with limited blood-brain-barrier penetration may maintain the antiobesity efficacy of rimonabant without unwanted side effects. This strategy has become the new hot spot for the development of antiobesity drugs. In this paper, we review the recent progress in peripherally restricted CB1 receptor antagonist.

OBJECTIVETo investigate the effect of propofol at doses for different anesthesia depths on γ-aminobutyric acid (GABA) in different cerebral regions at propofol uptake equilibrium in dogs.

METHODSTwelve 12-18-month-old healthy hybrid dogs weighing 10-12 kg were randomly divided into light anesthesia group (n=6) and deep anesthesia group (n=6) with a single bolus dose of propofol (5.5 and 7.0 mg/kg, respectively) completed in 15 s followed by intravenous propofol infusion at a constant rate (55 and 70 mg·kg(-1)·h(-1), respectively). Blood samples (2 ml) were taken from the internal carotid artery and jugular vein to measure plasma propofol concentrations 50 min after the start of the infusion. The dogs were then sacrificed and tissues were taken from different brain regions and the cervical cord to measure GABA concentrations using high-pressure liquid chromatography (HPLC).

RESULTSThe plasma propofol concentrations in internal carotid artery and jugular vein were similar in both light anesthesia group (3.00 ± 0.31 and 3.10 ± 0.51 µg/ml, respectively, P>0.05) and deep anesthesia group (6.41 ± 0.05 and 6.40 ± 0.11 µg/ml, respectively, P>0.05). GABA concentrations in the brain regions were significantly higher in deep anesthesia group than in light anesthesia group (P<0.05). The dorsal thalamus and hypothalamus showed greater GABA variations [(83.83 ± 2.230%) and (85.83 ± 1.72)%] compared to other brain regions at different anesthesia depths (P<0.05).

CONCLUSIONSIn both groups, plasma propofol concentrations in the internal carotid artery and internal jugular vein reach equilibrium at 50 min of propofol infusion. The variation of GABA is associated with the anesthesia depth of propofol, and GABA variation in the dorsal thalamus and hypothalamus plays an important role in propofol anesthesia.

Anesthetics, Intravenous ; pharmacokinetics ; Animals ; Brain ; metabolism ; Dogs ; Female ; Male ; Propofol ; blood ; pharmacokinetics ; gamma-Aminobutyric Acid ; metabolism

OBJECTIVETo observe the regional distribution of propofol in canine spinal cord under noxious stimulation.

METHODSTwelve healthy hybrid dogs (12-18 months old, weighing 10-12 kg) were randomly divided into control group (n=6) and stimulation group (n=6). All the dogs were anesthetized with a single bolus dose of propofol (7 mg/kg) in 15 seconds followed by propofol infusion at a constant rate of 70 mg/kg/h via the great saphenous vein of the right posterior limb. In the stimulation group, the tails of the dogs were clamped for 5 min after 45 min of propofol infusion. Blood samples were taken from the internal carotid artery and internal jugular vein at 50 min after propofol infusion to detect plasma propofol concentrations by high-pressure liquid chromatography (HPLC). The dogs were then immediately sacrificed by decapitation and the frontal horn, posterior horn, intermediate zone, frontal funiculus, posterior funiculus and lateral funiculus of the spinal cord were dissected for determination of propol content by HPLC.

RESULTSThe plasma concentrations of propofol in the internal carotid artery and internal jugular vein were 5.07-/+0.23 and 5.03-/+0.10 microg/ml in the stimulation group, respectively showing no significant differences from those in the control group (5.09-/+0.03 and 5.08-/+0.03 microg/ml, P>0.05). In the control group, the propofol concentration was 5.09-/+0.08 microg/g in the frontal horm, 5.10-/+0.08 microg/g in the posterior horn, 5.05-/+0.19 microg/g in the intermediate zone, 5.06-/+0.14 microg/g in the frontal funiculus, 5.06-/+0.15 microg/g in the posterior funiculus and 5.06-/+0.41 microg/g in the lateral funiculus, showing no significant differences (P>0.05). The propofol concentrations in the frontal horn (7.65-/+0.47 microg/g) and posterior funiculus (7.06-/+0.82 microg/g) in the stimulation group were significantly higher than those in the other spinal cord tissues (P<0.05) and those in the control group (P<0.05).

CONCLUSIONAt 50 min after intravenous injection of propofol at a constant rate of 70 mg/kg/h, plasma propofol concentrations in the internal carotid artery and internal jugular vein reaches equilibrium with a balanced distribution in all the spinal cord regions. Propofol concentration can be higher in the frontal horn and posterior funiculus under noxious stimulation.

Animals ; Dogs ; Female ; Male ; Nociceptors ; drug effects ; physiology ; Pain ; physiopathology ; Physical Stimulation ; Propofol ; administration & dosage ; pharmacokinetics ; pharmacology ; Random Allocation ; Spinal Cord ; metabolism

OBJECTIVETo explore the mechanism of electroacupuncture therapy on Parkinson's disease (PD).

METHODSFifty Wistar rats were randomly divided into a normal group, a sham-operation group, a model group, a Fengfu-Taichong group and a Shuanggu Yitong group. PD model was duplicated by microinjection of 6-Hydroxyl-Dopamine into right corpora striata, and by microinjection of normal saline in sham-operation group. Rats in normal group, sham-operation group and model group were not treated. In Fengfu-Taichong group, the rats were treated by electroacupuncture at "Fengfu" (GV 16) and "Taichong" (LR 3) on the basis of the PD model, and by electroacupuncture at "Fengfu" (GV 16), "Taichong" (LR 3), "Guanyuan" (CV 4) and "Zusanli" (ST 36) in Shuanggu Yitong group, once daily for 2 weeks. GDNF and Ret expression were detected by immunohistochemistry and western blotting, respectively.

RESULTSThe number of GDNF positive cells and the content of Ret receptor increased significantly in the two electroacupuncture groups compared with those in the other groups (all P < 0.01), and the expression of GDNF increased significantly in Shuanggu Yitong group compared with that in Fengfu-Taichong group (P < 0.01).

CONCLUSIONElectroacupuncture can not only increase the expression of GDNF, but also enhance its effect. "Shuanggu Yitong" method is better than simple acupuncture at "Fengfu" (GV 16) and "Taichong" (LR 3) in increasing expression of GDNF.

Animals ; DNA-Binding Proteins ; genetics ; metabolism ; Disease Models, Animal ; Electroacupuncture ; Gene Expression ; Glial Cell Line-Derived Neurotrophic Factor ; genetics ; metabolism ; Humans ; Nuclear Proteins ; genetics ; metabolism ; Parkinson Disease ; genetics ; metabolism ; therapy ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effects of ulinastatin on lung injury in hemorrhagic shock rats.

METHODSTwenty-four normal SD rats were randomly divided into 3 groups (n=8), namely the control group, hemorrhagic shock group (group H) and ulinastatin group (group U). In group H and group U, blood was drawn from the femoral artery over a period of 10 min until a mean arterial pressure of 40 mmHg was obtained. Controlled hypotension was then maintained at 40-/+5 mmHg for 60 min by blood drawing or infusion when necessary. All the blood drawn and an equivalent volume of Ringer lactate solution were subsequently infused for resuscitation. Four hours after the resuscitation, the activity of superoxidedismutase (SOD), content of malondialdehyde (MDA), expression of heme oxygenase-1 (HO-1), wet to dry weight ratio (W/D), and pathologic changes of the lung tissues were measured or observed.

RESULTSCompared with those in the control group, the content of MDA, expression of HO-1 and W/D increased significantly in both group H and group U (P<0.05); these indexes in group U were significantly lower than those in group H (P<0.05). The activity of SOD in group U was significantly lower than that in the control group (P<0.05) but higher than that in group H (P<0.05). Optical microscopy demonstrated milder inflammatory cell infiltration and interstitial edema in the lung tissues in group U than in group H.

CONCLUSIONUlinastatin can lower the content of MDA, W/D and the expression of HO-1, increase the activity of SOD, and reduce histological lung injury in rats with hemorrhagic shock.

Animals ; Glycoproteins ; pharmacology ; Heme Oxygenase-1 ; metabolism ; Lung Injury ; etiology ; prevention & control ; Male ; Malondialdehyde ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Shock, Hemorrhagic ; complications ; metabolism ; Superoxide Dismutase ; metabolism

Objective To explore and compare the relevant regional anatomies as they relate the fron- tolateral keyhole approach under microscopy and neuroendoscopy for operations in anterior cranial base and sellar region.Methods Fifteen silieone-injected cadaveric heads were dissected to reveal and compare the extent of expesure through the transfrontolateral keyhole approach under neuroendoscopy and microscopy. Results Portions in the areas of olfactory groove,sellar region and sylvian tissure were blind under micro- scope.Endoscope could allow observation of areas considered blind under the microscope.It could increase light intensity during the approach to objects,extend viewing angles,clear depiction of details in close-up po- sitions and inspect hidden structures.But images of endoscope were two dimensional,lack of view depth.Mi- croscopy and neuroendoscopy could help each other to recuperate deficiency.Conclusion Endoscope-assis- ted neuromicrosurgery is helpful,safe and minimally invasive to treat deepseated lesions in anterior cranial base,sellar region by transfrontolateral keyhole approach.

OBJECTIVETo investigate the protective effect of reactive oxygen species (ROS) scavenger, N-acetyl-L-cysteine (NAC), against H9c2 cardiomyocytes from injuries induced by chemical hypoxia.

METHODSH9c2 cells were treated with cobalt chloride (CoCl2), a chemical hypoxia-mimetic agent, to establish the chemical hypoxia-induced cardiomyocyte injury model. NAC was added into the cell medium 60 min prior to CoCl2 exposure. The cell viability was evaluated using cell counter kit (CCK-8), and the intercellular ROS level was measured by 2', 7'- dichlorfluorescein-diacetate (DCFH-DA) staining and photofluorography. Mitochondrial membrane potential (MMP) of the cells was observed by Rhodamine123 (Rh123) staining and photofluorography, and the ratio of GSSG/ (GSSG+GSH) was calculated according to detection results of the GSSG kit.

RESULTSExposure of H9c2 cardiomyocytes to 600 micromol/L CoCl2 for 36 h resulted in significantly reduced cell viability. Pretreatment with NAC at the concentrations ranging from 500 to 2000 micromol/L 60 min before CoCl2 exposure dose-dependently inhibited CoCl2-induced H9c2 cell injuries, and obviously increased the cell viability. NAC at 2000 micromol/L obviously inhibited the oxidative stress induced by CoCl2, decreased the ratio of GSSG/(GSSG+GSH), increased ROS level, and antagonized CoCl2-induced inhibition on MMP.

CONCLUSIONNAC offers obvious protective effect on H9c2 cardiomyocytes against injuries induced by chemical hypoxia by decreasing in the ratio of GSSG/(GSSG+GSH) and ROS level and ameliorating MMP.

Animals ; Cell Hypoxia ; drug effects ; Cells, Cultured ; Embryo, Mammalian ; Free Radical Scavengers ; pharmacology ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Oxidative Stress ; drug effects ; Rats ; Reactive Oxygen Species ; metabolism