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A growing number of studies have identified sex differences in response to general anesthesia; however, the underlying neural mechanisms are unclear. The medial preoptic area (MPA), an important sexually dimorphic structure and a critical hub for regulating consciousness transition, is enriched with estrogen receptor alpha (ERα), particularly in neuronal clusters that participate in regulating sleep. We found that male mice were more sensitive to sevoflurane. Pharmacological inhibition of ERα in the MPA abolished the sex differences in sevoflurane anesthesia, in particular by extending the induction time and facilitating emergence in males but not in females. Suppression of ERα in vitro inhibited GABAergic and glutamatergic neurons of the MPA in males but not in females. Furthermore, ERα knockdown in GABAergic neurons of the male MPA was sufficient to eliminate sex differences during sevoflurane anesthesia. Collectively, MPA ERα positively regulates the activity of MPA GABAergic neurons in males but not in females, which contributes to the sex difference of mice in sevoflurane anesthesia.
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Animals , Female , Male , Mice , Anesthesia , Estrogen Receptor alpha/metabolism , Preoptic Area , Sevoflurane/pharmacology , Sex CharacteristicsABSTRACT
The lateral hypothalamic area (LHA) plays a pivotal role in regulating consciousness transition, in which orexinergic neurons, GABAergic neurons, and melanin-concentrating hormone neurons are involved. Glutamatergic neurons have a large population in the LHA, but their anesthesia-related effect has not been explored. Here, we found that genetic ablation of LHA glutamatergic neurons shortened the induction time and prolonged the recovery time of isoflurane anesthesia in mice. In contrast, chemogenetic activation of LHA glutamatergic neurons increased the time to anesthesia and decreased the time to recovery. Optogenetic activation of LHA glutamatergic neurons during the maintenance of anesthesia reduced the burst suppression pattern of the electroencephalogram (EEG) and shifted EEG features to an arousal pattern. Photostimulation of LHA glutamatergic projections to the lateral habenula (LHb) also facilitated the emergence from anesthesia and the transition of anesthesia depth to a lighter level. Collectively, LHA glutamatergic neurons and their projections to the LHb regulate anesthetic potency and EEG features.
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Objective To evaluate whether the thalamic paraventricular nucleus mediates orexiner-gic ( orexin ) neurons-induced promotion of emergence from general anesthesia by using the optogenetics method in mice. Methods Twenty healthy male Hcrt-cre mice, aged 8-10 weeks, weighing 20-25 g, were divided into 4 groups ( n=5 each) using a random number table method: retrograde labeled viruses channelrhodopsin group ( R group) , anterograde labeled viruses channelrhodopsin group ( A group) , retro-grade labeled viruses control group ( RC group ) , and anterograde labeled viruses control group ( AC group) . The optogenetics technique was used in each group. Anesthesia was induced and maintained through inhaling 1％ isoflurane and pure oxygen 1. 0 L∕min. Electroencephalogram was monitored througout the procedure with the PowerLab monitoring system. The burst suppression ratio ( BSR) was recorded at 1 min before light stimulation and during light stimulation. Results Compared with RC group or the baseline at 1 min before light stimulation, the BSR was significantly decreased during light stimulation in R group ( P<0. 05) . Compared with AC group or the baseline at 1 min before light stimulation, the BSR was signifi-cantly decreased during light stimulation in group A ( P<0. 05) . Conclusion Optogenetics technique ap-plication once again confirms that orexin neurons can promote emergence from general anesthesia through thalamic paraventricular nucleus in mice.
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Postmenopausal osteoporosis is a type of osteoporosis with high bone transformation rate, caused by a decrease of estrogen in the body, which is a systemic bone disease characterized by decreased bone mass and increased risk of fracture. In recent years, as a kind of non-pharmacologic treatment of osteoporosis, defined by whole-body vibration less than 1 ( = 9.81 m/s ), low magnitude whole-body vibration is widely concerned, mainly because of its small side effects, simple operation and relative safety. Studies have shown that low magnitude whole-body vibration can improve bone strength, bone volume and bone density. But a lot of research found that, the therapeutic effects of low magnitude whole-body vibration are different depending on ages and hormone levels of subjects for animal models or human patients. There has been no definite vibration therapy can be applied to each subject so far. Studies of whole-body and cellular level suggest that low magnitude whole-body vibration stimulation is likely to be associated with changes of hormone levels and directed differentiation of stem cells. Based on the analysis of related literature in recent years, this paper made a review from vibration parameters, vibration effects and the mechanisms, to provide scientific basis and clinical guidance for the treatment of postmenopausal osteoporosis with low magnitude whole-body vibration.
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Objective To provide new experimental evidences associated with the mechanisms of inhaled anesthetics, the effects of sevoflurane on the electric activities of inhibitory interneurons in basal forebrain area (BF) were observed.Methods C57BL/6 mice, aged 2-3 weeks, were used and BF sections were cut for whole patch-clamp recording.Artificial cerebrospinal fluid containing sevoflurane was given and action potential, inhibitory postsynaptic potential were recorded.Results Sevoflurane could increase the frequency of firing rate of inhibitory interneurons in basal forebrain area (P<0.001), which could increase the frequency of action potential caused by depolarization current (P<0.05), and increase the frequency of spontaneous inhibitory postsynaptic currents of pyramidal neurons (P<0.05), while AP-depended miniature inhibitory postsynaptic currents were not significantly changed.Conclusion The basal forebrain inhibitory interneurons are involved in the anesthetic effect of sevoflurane.
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Objective To understand the health status of medical staffs in a tertiary hospital of Chongqing City to provide the basic information and scientific basis for formulating the interventional measures.Methods The physical examination data of 1 397 medical staffs in 2013 and 2 178 medical staffs in 2015 were selected as the research objects and performed the summary analysis according to the abnormal indexes distribution,sex and age.Results A total of 1 397 persons completed the physical examination in 2013,the abnormal indexes were detected in 1 250 people,the detection rate of sub-health status was 89.48％;2 178 people in 2015 completed the physical examination,the abnormal indicators were detected in 2 046 people,the detection rate of sub-health status was 93.94％.The comparison of two-year health examination data showed the different degrees of increasing trend in bilateral breast hyperplasia,gallbladder polyps,thyroid cysts,low density lipoprotein cholesterol.Conclusion The health status of medical staffs is not optimistic,so the self health care consciousness of medical staffs needs to be elevated.
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Objective To discuss the risk factors of C5 never palsy incidence after cervical open-door laminoplasty. Methods This study involved 254 patients with cervical spondylotic myelopathy, who underwent expansive open-door lam?inoplasty in our hospital between January 2009 and December 2012. Patients were divided into two groups according to the incidence of C5 palsy after operation. Group A included 20 patients with C5 palsy and group B included 234 cases without C5 palsy. The risk factors for postoperative C5 palsy were compared between two groups. The factors may affect the inci?dence of C5 palsy were analyze. Results All cases received follow-up with an average follow-up time (24.9±2.6) months. Values of cervical curvature index (CCI), lamina open angles and spinal cord shift were significantly greater in group A than those of group B (P0.05). Results of Logistic regres?sion analysis of multiple factors showed that lamina open angles, the pathogenic factors of C5 palsy included the spinal cord shift and postoperative CCI [OR(95%CI):2.035(1.684-2.359), 3.542 (2.465-9.254) and 9.126(4.425-17.273). Conclusion The angle of open door too large, CCI and spinal cord shift are risk factors for postoperative C5 palsy.
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Objective To evaluate the effect of dexmedetomidine on synaptic transmission in the spinal dorsal horn of rats.Methods Male Sprague-Dawley rats,aged 4-6 weeks,weighing 150-200 g,were used in the study.The lumbar enlargemnent segments of the spinal cord were harvested,and the parasagittal lumbosacral spinal cord slices with attached dorsal roots were prepared and incubated in artificial cerebro-spinal fluid.The whole-cell patch-clamp technique was used to record each index,and 4 spinal cord slices were selected and used for each index records.Experiment Ⅰ Dexmedetomidine was added cumulatively in concentration increments.Aδ and C fibers-mediated evoked excitatory postsynaptic currents (eEPSCs) were recorded before administration (baseline) and during perfusion with dexmedetomidine 4 and 10 μg/ml.Experiment Ⅱ The neurons innervated by Aδ and C fibers were selected,and Aδ and C fibers-mediated eEPSCs were recorded before administration (baseline),at 5 min of perfusion with yohimbine (alpha 2 adrenergic receptor antagonist) 2 μmol/L,and during continuous perfusion with yohimbine 2 μmol/L plus dexmedetomidine 4 μg/ml.Experiment Ⅲ The evoked excitatory postsynaptic potentials (eE-PSPs) and evoked inhibitory postsynaptic potentials (eIPSPs) were recorded before administration (baseline) and during perfusion with dexmedetomidine 4 μg/ml.Results Dexmedetomidine could dose-dependently inhibit Aδ and C fibers-mediated eEPSCs,dexmedetomidine could inhibit Aδ and C fibers-mediated eEPSPs and produced no effect on eIPSPs,and yohimbine could inhibit dexmedetomidine-induced inhibitory effect on eEPSCs.Conclusion The mechanism by which dexmedetomidine inhibits nociceptive information transmission in the spinal dorsal horn is related to inhibition of excitatory synaptic transmission through activating α2-adrenergic receptors,but not related to activation of inhibitory synaptic transmission in rats.
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Objective To explore the efficacy of four-step method for transurethral enucleation of the prostate . Methods A retrospective analysis was conducted on 120 cases of benign prostate hyperplasia ( BPH) treated with four-step method ( traction, enucleation, resection, and repair ) for transurethral plasmakinetic resection of the prostate in our hospital from January 2012 to February 2014. Results The average operation time was (91.4 ±35.3) min.The intraoperative blood loss was 78.8 ±34.1 ml. The resection of the prostate tissue weight was (32.1 ±15.7) g.The postoperative bladder infusion time was (1.5 ±0.5) days.The postoperative indwelling catheter time was 3.5 ±0.8 days.The postoperative hospital stay was (5.5 ±0.5) days.No death, blood transfusion, or TURS happened.Follow-up reviews at 3 months after surgery found postoperative international prostatic symptom score (IPSS) was significantly less than the preoperative level [4.5 ±2.2 vs.24.5 ±5.1, t=39.445, P=0.000], postoperative Qmax was significantly higher than preoperative level [(21.5 ±10.5) ml/s vs.(6.0 ±1.9) ml/s, t =15.912, P =0.000], and postoperative residual urine was significantly less than the preoperative level [(12.0 ±2.0) ml vs.(145.0 ±33.0) ml;t=44.069, P =0.000].No retrograde ejaculation, sexual dysfunction, or dysuria happened. Conclusion The four-step method for transurethral enucleation of the prostate is radical , safe, and simple.
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Bone marrow-derived mesenchymal stem cells (BMSCs) for repairing damaged heart tissue are a new kind of important treatment options because of their potential to differentiate into cardiomyocytes. We in this experiment investigated the effect of different electrical stimulation time on the expression of myocardial specificity gene and protein in rat bone marrow mesenchymal stem cells (rBMSCs) in vitro. The rBMSCs of second or third generation were randomly divided into three groups, i.e, electrical stimulation (ES) group, 5-Azacytidine (5-Aza) group and the control group. The rBMSCs in the ES groups with complete medium were exposed to 2 V, 2 Hz, 5 ms electrical stimulation for 0. 5 h, 2 h, 4 h, and 6 h respectively every day for 10 days. Those in the 5-Aza group were induced by 5-Aza (10 μmol/L) for 24 h, and then cultured with complete medium for 10 days. Those in the control group were only cultured with complete medium, without any treatment, for 10 days. The rBMSCs' morphological feature in each group was observed with inverted phase microscope. The mRNA expression of myocyte-specific enhancer factor 2C (MEF-2C) and connexin 43 (Cx43) were examined with Real-Time quantitative PCR and the protein expression of MEF-2C, Cx43 were detected with Western Blot method. The results showed that the mRNA expression level of the MEF-2C, Cx43 and the protein expression level of MEF-2C, Cx43 were significantly higher in the ES group and 5-Aza group than those in the relative control group (P < 0.05). It suggests that electrical stimulation could play a part of role in the induction of the rBMSCs to differentiate into the cariomyocyte-like cells in vitro and the effectiveness of the electrical stimulation with 2 h/d had the best in our experiment. But the mechanism how electrical stimulation promotes the differentiation of rBMSC into cardiomyocyte is still unclear.
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Animals , Rats , Biomarkers , Metabolism , Cell Differentiation , Cells, Cultured , Connexin 43 , Metabolism , Electric Stimulation , MEF2 Transcription Factors , Metabolism , Mesenchymal Stem Cells , Cell Biology , Metabolism , Myocytes, Cardiac , Cell Biology , RNA, Messenger , Metabolism , Rats, Sprague-DawleyABSTRACT
Objective To evaluate the therapeutic effect of ureteroscopy with holmium laser on ureteral stricture associated with calculi.Methods The clinical data of 36 cases of ureteral stricture associated with calculi undergoing ureteroscopy with holmium laser treatment were analyzed retrospectively with 26 cases of male patients and 10 cases of female patients.The age was 26-65 years.There were upper and middle ureteral stricture associated with calculi in 8 cases,and lower segment of ureteral stricture associated with calculi in 28 cases.All the patients were treated with ureteroscopy with holmium laser.All cases were followed up for 6-12 months.In the follow-up of 36 cases B ultrasound and urinary tract flat piece or intravenous urinary tract imaging were performed to evaluate the therapeutic effect.Results Thirty-six patients underwent ureteroscopy with holmium laser successfully.No conversion to open surgery occurred.Mean operating time was 30 (20-45) min.Mean hospital stay postoperatively was 3-5 d.Postoperative 8 cases with low thermal,alleviated by the anti-infection treatment,no ureteral avulsion,leakage and severe complications such as hemorrhage.Postoperative follow-up,32 cases (89％,32/36) were cured and 4 cases (11％,4/36) were improved.Conclusion Ureteroscopy with holmium laser in treatment of ureteral stricture associated with calculi is a simple and effective minimally invasive method.
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Objective To investigate the practical efficacy and safety of ultrasound with microbubbles mediated rAAV2-EGFP to retina of rat after intravitreal and subretinal injection. Methods Gene transfer was examined by rAAV2-EGFP intravitreal and subretinal injection into the Wistar rats with or without microbubbles. The eyes were exposed to US (1 MHz,2 W/cm2, duration 5 minutes,duty cycle 50%,pulse recurrent frequency 100 Hz). The onset of EGFP gene expression, lightness of fluorescence, area of fluorescence and its distribution in the fundus in vivo via fluorescence stereosocope were investigated on the 4th,7th, 35th,49th and 120th day respectively. The value of gene transfer was quantified through the EGFP fluorescence quantitative methods by Axiovision 3. 1 software. HE staining was used to observe tissue damage. Results There was no fluorescence observed by fluorescence stereosocope after intravitreal injection after two-month study. After subretinal injection, ultrasound-targeted microbubbles destruction (UTMD) strongly increased gene transfer efficiency. UTMD used in the experiment did no harm to the rat retina structure. Conclusions UTMD could not enhance rAAV2-EGFP transfecion efficiency to rat retina after intravitreal injection but the transduction could be enhanced significantly after subretinal injection.
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Objective To investigate changes of metabolism of glucose and lipoids in diabetes before and after Ramadan. Methods A total of 65 Muslim fasting diabetic patients, with complete data, were investigated about their diet and administrated medicines before and after the fasting and occurrence of hypoglycemia during the fasting. Indexes of WEIGHT, WAISTLINE, BMI, FPG, 2h-PG, HbA1c, TG, TC, HDL-C, and LDL-C were evaluated by questionnaire, physical examination and laboratory tests. Results During Ramadan, fasting time was prolonged (P＜0.01), intake of lipoids and protein were increased obviously (P＜0.01) and a significant increase of 2 h-PG, HbA1c (P=0.0149, 0.0175) was detected after Ramadan in s fasting group. No significant difference was found in WEIGHT, WAISTLINE, BMI, TG, TC, HDL-C, and LDL-C. Blood glucose was improved significantly in the control group than the fasting group. Conclusion Life style was changed obviously leading to unsatisfactory controlling of blood glucose in Muslim diabetic patients during Ramadan.
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Objective To investigate the changes of sex hormone and androgen receptor levels and evaluate the relationship of the sex hormones and androgen receptor with coronary heart disease (CHD) in elderly men. Methods A cross-sectional study was performed in 539 elderly men, including 400 healthy people aged 62-92 years and 139 CHD patients aged 60-88 years. The plasma concentrations of total testosterone (TT), free testosterone (FT), dehydroepiandrosterone sulfate (DHEAS), sex hormone binding globulin (SHBG), estradiol (E2), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were measured. The androgen receptor (AR) level was tested by flow cytometry. Results The fluorescence intensity of DHEAS, TT, SHBG, FT and AR were significantly lower in CHD group than in healthy group (P<0.01);however, FSH and E2 in CHD group were higher(P(0.01). Age was negatively correlated with TT(r=-0.28,P<0.01) and FT (r=-0.17,P<0.05), and positively correlated with SHBG(r=0.14,P<0.05) and E2 (r=0.33, P<0.01). AR fluorescence intensity was negatively correlated with systolic blood pressure (r=-0.12,P<0.01). Logistic regression analysis indicated that TT (OR=1.065,9% CI: 1.012～1.121,P<0.05), SHBG(OR=0.994,95% CI:0.990～0.998,P<0.01) and AR (OR=0.971,95%CI:0.956～0.986, P<0.01)were significantly associated with CHD in elderly male patients. Conclusions The levels of DHEAS, TT, SHBG, FT and AR are lower in elderly men with CHD than in elderly healthy men;however, the FSH and E2 concentrations are higher. Low levels of TT, SHBG and AR may be the independent risk factors for CHD in elderly men.
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A fusion gene called Ig-BDNF, in which brain-derived neurotrophic factor cDNA fused to the 3' end of signal peptide of Ig coding sequence, was constructed by PCR, digested and subcloned into shuttle plasmid pSNAV to obtain a recombinant plasmid pSNAV-Ig-BDNF. Then the plasmid encoding fusion protein was transfected into 293 cell lines and the stably transfected clones were selected with neomycin. AAV1 containing Ig-BDNF fusion gene vectors were obtained by super-infection by Herpes virus. The resultant adeno-associated virus vectors AAV-Ig-BDNF were confirmed by PCR, Western blotting and a sandwich enzyme-linked immunosorbent assay (ELISA) after infection of 293 cell lines. The results indicated that AAV-Ig-BDNF contained the target gene, and infected cells and produced the fusion protein into the supernatant. The content of BDNF in medium per 5x104 cells over a 24 h incubation period reached 1000 pg/mL. With the help of non-replicative adenovirus during AAV-Ig-BDNF infection, the expression of BDNF increased 7-8 fold, and the enhancement of BDNF gene expression was observed in a concentration-dependent manner. These results suggested that a functional AAV-Ig-BDNF was successfully constructed and it offers basis for further study for gene therapy of neural degeneration diseases.
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Humans , Brain-Derived Neurotrophic Factor , Genetics , Cell Line , Dependovirus , Genetics , Metabolism , Genetic Vectors , Genetics , Recombinant Fusion Proteins , Genetics , Recombination, Genetic , Simplexvirus , Genetics , TransfectionABSTRACT
Objective To investigate the expression and characteristics distribution of ciliary neurotrophic factor (CNTF) and its receptor during the development of retina of healthy Sprague-Dawley(SD)and Royal College of Surgeons (RCS) rats with hereditary retinal degeneration. Methods The expression and distribution of ciliary neurotrophic factor and its receptor were detected by immunohistochemical staining in the retinal paraffin sections of SD and RCS rats from newborn to 12 moths old. Results In the normal retina of SD rats 0-7 days after birth, positive CNTF staining was found in all of the retinal layers and the staining of ganglion cells strengthened and other cells weakened as the age of rats increased; the staining of ganglion cells reached the peak at the 4th week and lasted till the agedness. The same results of the CNTF staining were also found in RCS rats retina. Weak positive staining of CNTFR in all of the retinal layers was seen in the 0-3-day-old SD rats; the ganglion cells were darkly stained and incontinuous positive staining at the site which would develop to be the external segment was found; as the age increased, the positive staining of external segment of photoreceptor enhanced and reached the peak at the 14-28th day after birth. At the 56th day, the staining of ganglion cells in retina of SD rats was strengthened while the staining of external segment weakened till the agedness. The expression of CNTFR in retina of 3-14-day-old RCS rats was the same as which of normal SD rats basically, but the staining of external segment weakened obviously from the 21st day on, and negative staining of external and positive ganglion cells were detected at the 28th day till the agedness. Conclusions Expression of CNTF in normal SD rats and RCS rats with hereditary retinal degeneration is almost the same. The presence of significant difference of expression of CNTFR between normal SD rats retina and RCS rats retina may provide the experimental gist for the CNTF treatment to retinal degeneration.
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The pathogenesis and clinical manifestation of histiocytosis are veriant, and misdiagnosis is relatively high. We reviewed a total of cases of this disease and related literature, and the possibility pathogenesis and its clinical features. were discussed.