A Novel Retrograde AAV Variant for Functional Manipulation of Cortical Projection Neurons in Mice and Monkeys / 神经科学通报·英文版

Retrograde adeno-associated viruses (AAVs) are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks. However, few retrograde AAV capsids have been shown to offer access to cortical projection neurons across different species and enable the manipulation of neural function in non-human primates (NHPs). Here, we report the development of a novel retrograde AAV capsid, AAV-DJ8R, which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques. In addition, intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations. Moreover, AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen. These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations.

Treatment Outcome of Acute Sacral Nerve Stimulation in Functional Anorectal Pain.

BACKGROUND: Sacral nerve stimulation (SNS) has revolutionized the management of certain intractable cases of fecal and urinary incontinence; however, the management of functional anorectal pain (FAP) has been addressed in only a few studies. OBJECTIVE: The aim of this study was to evaluate the treatment effect of SNS in improving FAP symptoms. METHODS: A total of 120 patients with FAP who had undergone temporary SNS probe placement were investigated at Qianfoshan Hospital between January 2014 and December 2016. Pre- and post-SNS treatment outcomes were assessed using the VAS, anorectal manometry, and the 36-item short-form health survey (SF-36) medical outcomes study instrument. RESULTS: A total of 120 patients proceeded to insertion of an SNS probe at the S3 nerve root (2 Hz, 1.50 mA, 0.10 milliseconds). Of these, 75 patients were cured, 41 improved, and 4 had an ineffective outcome. The total effectiveness rate was 96.7% 1 year after treatment. There was a significant reduction in the median VAS score pre-SNS and post-SNS, from 8 to 3, respectively. Patients post-SNS had lower anal maximum contraction pressure and anal rest pressure than did patients pre-SNS. Compared with the pretreatment group, there were no substantial differences between anal longest contraction time and rectal rest pressure. In addition to general health, there was a substantial improvement in the remaining dimension scores of the SF-36. CONCLUSION: The effect of SNS in treating FAP was positive, and the improvement of symptoms was substantial and worthy of clinical promotion.

Magnetic resonance imaging study on the correlation between changes in the puborectalis muscle and posterior perianal abscess / 中华普通外科杂志

Objective To investigate the correlation between posterior perianal abscess and thickness of the puborectalis muscle by magnetic resonance imaging.Methods Sixty-three patients with a low-position posterior perianal abscess (the low-position group),44 patients with a high-position posterior perianal abscess (the high-position group) and 46 healthy volunteers (the control group) were enrolled in this study.In the lithotomy position,MRI was performed to measure the thicknesses of the puborectalis muscle at the 5,6 and 7 o'clock positions of the rectum.Results The thickness of puborectalis muscle at 5,6 and 7 o'clock points in the control group was (5.5 ±0.8),(9.0 ±0.7),(5.4 ±0.6) mm,while that in the low-position group was (5.4 ± 0.7),(10.0 ± 0.9),(5.5 ± 0.7) mm,and that in the high-position group was (7.3 ± 1.0),(11.7-1.2),(7.5 ±0.9) mm,respectively.The thicknesses of the puborectalis muscle at the 5,6,and 7 o'clock positions of the rectum in the lithotomy position were compared between the control group and the low-position group.The difference in the thickness at the 6 o'clock position was statistically significant(t =6.492,P ＜ 0.05),but there were no significant differences at 5 and 7 points (t =-0.719,0.633,P＞0.05).The thicknesses of the pubqrectalis muscle at the 5,6 and 7 o'clock positions of the rectum in the lithotomy position were compared between the control group and the high-position group,and the differences were statistically significant at each position (t =9.476,12.875,13.649,P ＜ 0.05).Simultaneously,the differences between the low-position group and the high-position group were statistically significant(t =-11.206,-7.835,-13.361,P ＜ 0.05).Conclusions Magnetic resonance imaging can diagnose perianal abscess in different parts by measuring the thickness of puborectal muscle,which provides strong evidence for early intervention treatment,so as to improve the therapeutic effect and reduce post-operative anal fistula.

Clinical Observation of Dezocine and Nalbuphine on Patient-controlled Intravenous Analgesia in Patients Undergoing Cesarean Section / 中国药房

OBJECTIVE:To compare the effects of dezocine and nalbuphine on patient-controlled intravenous analgesia(PCIA) in patients undergoing cesarean section. METHODS:A total of 97 patients undergoing selective cesarean section were selected from our hospital during Jun. 2015 to Mar. 2017. They were divided into dezocine group(52 cases)and nalbuphine group(45 cases) according to lottery. Both groups received cesarean section under combined spinal-epidural anesthesia,and then given PCIA pump immediately after surgery. The pump of dezocine group was Dezocine injection 0.5 mg/kg+Tropisetron hydrochloride injection 10 mg;that of nalbuphine group was Nalbuphine hydrochloride injection 2 mg/kg+Tropisetron hydrochloride injection 10 mg. Both groups of analgesic drugs were diluted 100 mL with 0.9% sodium chloride injection,constant infusion of liquid medicine at rate of 2 mL/h,adding 0.5 mL additionally each time,for consecutive 48 h. VAS score and Ramsay sedation score of resting pain, dynamic pain and uterine contraction pain were performed in 2 groups 4,8,12,24,48 h after surgery. The serum levels of PRL were determined 30 min before surgery and 24,48 h after surgery. The initial time of lactation and ADR were recorded in 2 groups. RESULTS:VAS score of resting pain and uterine contraction pain at 4,8,12 h after operation and that of dynamic pain at 4,8,12, 24 h after operation were significantly lower in dezocine group than nalbuphine group,with statistical significance (P＜0.05). There was no statistical significance in VAS score between 2 groups at other time points(P＞0.05). As time went on,the VAS scores of the two groups decreased significantly at each time point,and the difference was statistically significant(P＜0.05). The serum levels of PRL in 2 groups 24 and 48 h after operation were significantly higher than 30 min before operation,with statistical significance(P＜0.05). There was no statistical significance in serum level of PRL between 2 groups at same time point(P＞0.05). There was no statistical significance in Ramsay score, initial time of lactation or the incidence of ADR between dezocine group and nalbuphine group (P＞0.05). CONCLUSIONS:Both dezocine and nalbuphine are effective analgesia drugs of PCIA in patients undergoing cesarean section. Early postoperative analgesic effect of dezocine is superior to nalbuphine. They have similar effects on long-term analgesia and postoperative sedative,serum level of PRL,initial time of lactation,as well as safety.

Accelerated decline in anti-HCV antibody levels in spontaneous HCV seroconverters with HIV co-in-fection / 中华微生物学和免疫学杂志

Objective To investigate the influence on anti-HCV antibody levels in spontaneous HCV seroconverters co-infected with HIV. Methods A retrospective study was conducted on people with a history of blood donation in Wangying Village,Shangcai County,Henan Province in 2009 and 2017. Accord-ing to the infection status in 2009,patients who were positive for anti-HCV antibody were divided into four groups:HIV-negative chronic HCV infection group (HCVc),HIV-negative spontaneous HCV clearance group (HCVr),HIV-positive chronic HCV infection group (HIV+HCVc),HIV-positive spontaneous HCV clear-ance group ( HIV+HCVr). All patients were followed up in 2017 and those who were lost to follow-up, received HCV treatment or were reinfected with HCV (only for those of HCV seroconverters) were excluded from this study. Altogether 167 patients met the inclusion criteria (HCVc:n=65;HCVr:n=34;HIV+HCVc:n=44;HIV+HCVr:n=24). A horizontal comparison of anti-HCV antibody levels among the above four groups in 2009 and a longitudinal comparison of changes in anti-HCV antibody in each group from 2009 to 2017 were respectively conducted. Results The horizontal comparison indicated that the levels of anti-HCV antibody were higher in chronic HCV-infected patients than in HCV seroconverters no matter whether they were co-infected with HIV or not (both P<0. 000 1). After comparison of anti-HCV antibody titers in 2017 and 2009,no significant changes were found in HCVc or HIV+HCVc group. The levels of anti-HCV antibody in HCVr and HIV+HCVr groups decreased significantly from 2009 to 2017 ( both P<0. 000 1). HIV+HCVr group showed a faster decline in anti-HCV antibody level than HCVr group (P=0. 003 9). Significant nega-tive correlations between the decline speed in anti-HCV antibody sample/cut-off ( S/CO) values and the initial anti-HCV antibody S/CO values (in 2009) were found in both HCVr (r=-0. 517 7, P=0. 001 7) and HIV+HCVr groups (r=-0. 753 2, P<0. 000 1). The decline speed in anti-HCV antibody in HIV+HCVr patients was found to be negatively correlated with their CD4+T cell counts in 2009 ( r=-0. 563 8, P=0. 004 1). Moreover,the seroreversion rate of anti-HCV antibody in patients of the HIV+HCVr group was higher than that of HCVr group (P=0. 027 5). Conclusion HIV co-infection can accelerate the decline of anti-HCV antibody in spontaneous HCV seroconverters. This study indicates that in a large-scale retrospective epidemiological investigation especially for HIV-infected populations, the prevalence of anti-HCV antibody may be underestimated.

Intracellular cytokine staining (ICS) for detection of NK cell-mediated antibody-dependent cell-mediated cytotoxicity (ADCC) / 中华微生物学和免疫学杂志

Objective To analyze the possibility of using intracellular cytokine staining (ICS) to evaluate NK cell-mediated antibody-dependent cellular cytotoxicity (ADCC) and to detect the changes in ADCC activity among patients with chronic HIV and/or HCV infection. Methods Flow cytometry was per-formed to determine the percentages of NK cells and the expression of NK cell receptors. ImageStreamX MarkⅡ system was used to identify the expression of CD3, CD56, CD16 and CD32 on CD56brightNK and CD56dimNK subsets. Degranulation process and cytokine production in NK cells were detected using an anti-gen-antibody complex model of P815/Ab in combination with ICS. Differences in NK cell-mediated ADCC were evaluated among patients infected with HIV and/or HIV and healthy subjects by flow cytometry. Re-sults The percentages of CD107a+and IFN-γ+NK cells were positively correlated with the decrease of mean fluorescence intensity (MFI) of CD16. ICS assay revealed a positive correlation between the secretion of CD107a and IFN-γ by NK cells. CD16 was highly expressed in CD56dimNK cells. The ADCC mediated by CD56dimNK cells was stronger than that mediated by CD56brightNK cells. The rate of target cell lysis detected by rapid fluorescence assay was positively correlated with the percentage of CD107a+/IFN-γ+NK cells meas-ured by ICS. NK cell-mediated ADCC was suppressed in patients with chronic HIV and/or HCV infection. Conclusion This study suggests that ICS assay could be used to evaluate NK cell-mediated ADCC. It also reveals that NK cell-mediated ADCC is suppressed in patients with chronic HIV and/or HCV infection.

Dyschesia and changes of pubo-rectal angle as observed by 3D ultrasound / 中华普通外科杂志

Objective To discuss the relativity of dyschesia with the change of puborectalismuscle.Methods 68 patients with dyschesia were compared with 68 healthy volunteers at lithotomy position undergoing 3D ultrasonography on resting,contracting and maximum exertion phase respectively.Three dimensionally reconstructed images were reconstructed and pubo-rectal angle and the thickness of puborectalis muscle at 6 o'clock position were measured.Results The pubo-rectal angle of the study group and control group had no significant differences at either resting [(86 ± 8) ° vs.(86 ± 8)°] or contracting phases [(88 ± 9) ° vs.(86 ± 7) °] (t =-0.145,t =0.434,P ＞ 0.05).While at maximum exertion the differences were significant [(80 ± 6) °vs.(95 ± 5) °,t =-5.397,P ＜ 0.05].The d-value of pubo-rectal angle between maximum and resting exertion statistically different [(6 ± 3) °,(-9 ± 7) °,t =5.551,P ＜ 0.05].The thickness of puborectalis muscle between the two groups differed statistically significant only at maximum exertion phase [(4.60 ± 0.60) mm vs.(3.97 ± 0.32) mm,t =6.872,P ＜ 0.05].The d-value of the thickness of puborectalis muscle between maximum and resting exertion were statistically different (t =-11.474,P ＜ 0.05).Conclusions The pubo-rectal angle of the study group is smaller at maximum exertion than the control group.The thickness of puborectalis muscle at six o'clock (lithotomy position) in study group is larger at maximum exertion than the control group.And the severity of constipation changed with the variation of angle.

Effect of astragaloside Ⅳ on postoperative cognitive function in aged rats / 中华麻醉学杂志

Objective To evaluate the effect of astragaloside Ⅳ on postoperative cognitive function in aged rats.Methods Sixty healthy male Sprague-Dawley rats,aged 22 months,weighing 360-480 g,were divided into 4 groups(n=15 each)using a random number table:control group(group C),surgery group(group S),low-dose astragaloside Ⅳ group(group L-AGS)and high-dose astragaloside IV group(group H-AGS).At 3 days prior to surgery,astragaloside Ⅳ 20 and 40 mg/kg were injected intraperitoneally once a day in L-AGS and H-AGS groups,respectively.The equal volume of normal saline was given instead in C and S groups.The animals underwent splenectomy under anesthesia with 1.8% isoflurane in S,L-AGS and H-AGS groups.Five rats in each group were randomly sacrificed at 1 day after operation,the hippocampi were removed for determination of interleukin-1beta(IL-1β),tumor necrosis factor-alpha(TNF-α)and IL-6 contents(by enzyme-linked immunosorbent assay)and expression of activated caspase-3,Bax and Bcl-2(by Western blot).The left animals underwent Morris water maze test at 15 days after operation.Results Compared with group C,the escape latency was significantly prolonged,the frequency of crossing the original platform was reduced,the space exploration time was shortened,the expression of activated caspase-3 and Bax was up-regulated,the expression of Bcl-2 was down-regulated,and the IL-1β,TNF-α and IL-6 contents were increased after operation in group S(P<0.05).Compared with group S,the escape latency was significantly shortened,the frequency of crossing the original platform was increased,the space exploration time was prolonged,the expression of activated caspase-3 and Bax was down-regulated,the expression of Bcl-2 was up-regulated,and the IL-1β,TNF-α and IL-6 contents were decreased after operation in L-AGS and H-AGS groups(P<0.05).Compared with L-AGS,the escape latency was significantly shortened,the frequency of crossing the original platform was increased,the space exploration time was prolonged,and the TNF-α contents were decreased after operation in group H-AGS(P<0.05).Conclusion Astragaloside Ⅳ can improve the postoperative cognitive function in a dose-dependent manner in aged rats.

Effect of sufentanil postconditioning on acute lung injury induced by ischemia-reperfusion of uterine in rats / 临床麻醉学杂志

Objective To investigate the effect of sufentanil postconditioning on acute lung in-jury induced by ischemia-reperfusion of uterine in rats.Methods Fourty-five adult female Sprague-Dawley rats were randomly divided into 3 groups(n = 1 5 each):control group (group C),ischemia-reperfusion group (group IR)and sufentanil postconditioning group(group SPC).Group SPC received sufentanil 10 μg/kg via intraperitoneal injection before inducing reperfusion of uterine.Ischemia-reper-fusion of uterine was produced by occlusion of bilateral uterine arteries for 45 min followed by reper-fusion for 2 h in group IR and group SPC.Then the content of tumor necrosis factor-α(TNF-α),mal-odiadehyde (MDA)and superoxide dismutase (SOD)activity were measured in uterus,serum and lung tissue,lung wet to dry weight ratio (W/D),lung permeability index (LPI)were compared. Results Compared with group C,TNF-α,MDA content,W/D and LPI in uterus,serum and lung tissue were significantly increased in group IR and group SPC(P <0.05).TNF-α,MDA content,W/D and LPI in uterus,serum and lung tissue were significantly attenuated in group SPC as compared with group IR (P <0.05 ).Compared with group C,SOD activity in uterus,serum and lung tissue was significantly attenuated in group IR and group SPC (P <0.05).SOD in uterus,serum and lung tissue were significantly increased in group SPC as compared with group IR (P < 0.05 ). Conclusion Sufentanil postconditioning attenuates pulmonary injury caused by ischemia-reperfusion of uterine in rats by inhibiting the inflammatory reaction and suppressing the activation of oxyradical.

Role of nicotinamide adenine dinucleotide phosphate oxidase in spinal cord in maintenance of diabetic neuropathic pain in rats / 中华麻醉学杂志

Objective To evaluate the role of nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase) in the spinal cord in the maintenance of diabetic neuropathic pain in rats.Methods Pathogenfree male Sprague-Dawley rats,aged 2 months,weighing 180-220 g,were used in the study.Diabetes mellitus was induced by intraperitoneal streptozotocin (STZ) 60mg/kg and confirmed by blood glucose ＞ 16.7 mmol/L at 72 h after STZ injection.Twenty diabetic rats were randomly allocated to diabetic neuropathic pain group (DN group,n =10) and apocynin (specific NADPH oxidase inhibitor) group (A group,n =10).Another 10 agematched normal rats served as control group (C group,n =10).Twenty-eight days after STZ injection,apyconin 5 mg/kg was injected intraperitoneally once a day for 7 consecutive days in A group.Paw withdrawal threshold to yon Frey filament stimulation (PWT) was measured before STZ injection (T1) and at 7,14,21,28 and 35 days after STZ injection (T2-6).The rats were sacrificed after PWT was measured at T6 and L4.5 segments of the spinal cord were removed for determination of NADPH oxidase subunits gp91phox and p47phox expression,MAD content and SOD activity.Results Compared with C group,PWT was significantly decreased at T3-5,gp91phox and p47phox expression was up-regulated,MAD content was increased,and SOD activity was decreased in DN and A groups.Compared with DN group,PWT was significantly increased at T6,gp91phox and p47phox expression was downregulated,MAD content was decreased,and SOD activity was increased in A group.Conclusion NADPH oxidase in the spinal cord is involved in the maintenance of diabetic neuropathic pain in rats.

To constructe,package and identificate the lentiviral vector with overexpression gene Grp78 / 重庆医学

Objective To constructe ,package and identificate the lentiviral vector with overexpression gene Grp78 .Methods We used lentiviral vector and genetic engineering technology to obtain the aim gene fragment and to constructe recombinant plas‐mid .we prepared competent cells and transform the cells .Through positive clone sequencing ,lentivirus was packaged and virus titer was tested .Results Positive cloning sequence comparison results show that the test was passed .Melt curve did not appear mixed peak ,also did not appear abnormal peak broadening .It means that does not appear pollution ,primer dimers and nonspecific amplifi‐cation in the experiments .Conclusion The construction ,packaging and identification of lentiviral vector with over expression gene Grp78 are sucessful .

Effects of sevoflurane on Ca2+ transsarcolemmal influx and Ca2+ release function of endoplasmic reticulum in isolated outer hair cells of guinea pigs / 中华麻醉学杂志

Objective To investigate the effects of sevoflurane on ca2+ transsarcolemmal influx and ca2+ release function of endoplasmic reticulum in isolated outer hair cells (OHCs) of guinea pigs and the possible mechanism by which sevofhlrane acts on cochleas.Methods The experiment was performed in 2 parts.In experiment I:twelve adult guinea pigs(8 male,4 female)weighing 180-230 g were used.OHCs were mechanically sparated after enzymatic incubation.Thirty OHCs with favorable activity were divided into 3 groups (n=10 each):group I control(C);group Ⅱ low concentration sevoflurane (1.7%,group S1) and group Ⅲ high concentration sevoflurane(3.4%,group S2).The OHCs were stained with 6 umol/L Fluo-3AM in estefified form for 40 min.Group S1 and S2 were pretreated with 1.7% and 3.4% sevoflugsne respectively for 20 min.KCI 40 mmol/L was then added.The intracellular ionized Ca2+ concentration ([C2+]I) was determined byintracelhlar Ca2+ fluorescent intensity using laser scanning confocal microscope.The protocol of the experimentⅡ was the same as the experimentI.The only difference was that caffeine 20 mmol/L was added instead of KCI 40 mmol/L.Results In experiment I:there was no significant difference in baseline[ca2+]I and[ca2+]I after being exposed to sevoflurane among the 3 groups.[Ca2+]I was significanfly increased after addition of KCI as compared with the baseline[Ca2+]I and was significantly lower in group Sl and S2 than in group C and was the lowest in group S2.In experimentⅡ:the[ca2+]I was significantly increased after addition of caffeine but there was no significant difference in[Ca2+]I among the 3 groups.Conclusion Sevoflurane can inhibit voltage-dependent Ca2+ channel opening in a concentration-dependent manner but can not affect ryanodine-sensitive Ca2+ release function of endoplasmic reticuhm in isolated outer hair cells of guinea pigs.

The Effect of Nitrous Oxide and Isoflurane on the Total RNA Yield from the Cochlea of the Rats / 华中科技大学学报（医学）（英德文版）

The possible mechanism of inhalation anesthetics on the internal auditory impairment of the rat was investigated by determining the effect of nitrous oxide (N20) and isoflurane on the total RNA yield from the cochlea of the rats. Thirty healthy Wistar rats were randomly divided into 3 groups: group C (control group, n=10) with a 3-h unremitting inhalation of 50% O2 group N (ex-periment group, n= 10) with a continuous inhalation of 50% N2O+50% O2for 3 h, and group I (ex-periment group, n=10) with a 3-h sustained inhalation of 2.5% isoflurane. The TRIzol in combination with RNeasy was used to respectively extract the total RNA from cochlea of rats in the 3 groups. Spectrophotometry was used to detect total RNA yield and electrophoresis to detect the quality. The total RNA extracted from the cochlea of the rats in the groups C and N was 7.69 and 6.51 μg, respec- tively. There was a 15% decrease in the N group as compared with group C. The total RNA from the rats in the group I was 7.32 μg, and there was hardly any change in the group as compared with the group C. The value of A260/A280 in groups C, N and I was 2.07, 2.04 and 2.04, respectively, showing a very high RNA purity. The result of gel electrophoresis suggested that there was no degradation in the total RNA. It was suggested that the interference of N2O on the cochlear RNA yield might be one of the reasons which cause an injury of the ear. The isoflurane shows no harm on the heating.

The changes in nitric oxide synthase activity and nitric oxide content in rat brain during different stages of enflurane anesthesia / 中华麻醉学杂志

Objective To determine the effect of enflurance on nitric oxide synthase (NOS) activity and nitric oxide (NO) content in different brain regions at different anesthesia stages.Methods Forty female SD rats weighing 250-350 g were randomly divided into five groups of 8 animals each, group 1: control; group 2: induction of anesthesia; group 3: maintenance of anesthesia; group 4: recovery from anesthesia and group 5: complete recovery. Animals were placed in a special glass anesthesia box. The concentration of enflurane in the box was measured by anesthesia gas monitor (Normac). The induction of anesthesia started from staggering of the animal to loss of righting reflex, maintenance of anesthesia from 1 min after loss of righting reflex, recovery of anesthesia from recovery of righting reflex to staggering after enflurance anesthesia was discontinued and glass box was opend and complete recovery from 1h after recovery of righting reflex. The animals were decapitated at different stages of anesthesia. Cerebral cortex, hippocampus and brain stem were immediately removed on ice and frozen in liquid nitrogen. Their NOS activity and NO content were measured by spectroscopic analysis. Results The average duration from the beginning of 2.2% enflurane inhalation to loss of righting reflex was (169?30) s. The average duration of anesthesia induction was (138?36)s. The maintenance of anesthesia lasted (229?30) s and the recovery period averaged (266?41) s. The NOS activity and NO content in different brain regions began to decrease during induction of anesthesia and reached their lowest level during maintenance of anesthesia, began to increase during recovery from anesthesia and returned to preanesthetic level during complete recovery. Conclusion Enflurane significantly inhibits the NOS activity and NO content in the brain. The inhibits is closely related to the depth of anesthesia. NO is a message transmitter in central nervous system and may be involved in the mechanism of enflurane anesthesia.