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.
Animals ; Haplorhini ; Axons ; Motor Neurons ; Interneurons ; Macaca ; Dependovirus/genetics* ; Genetic Vectors

Objective The study aims to investigate the effects of different adaptive statistical iterative reconstruction-V（ ASiR-V） and convolution kernel parameters on stability of CT auto-segmentation which is based on deep learning. Method Twenty patients who have received pelvic radiotherapy were selected and different reconstruction parameters were used to establish CT images dataset. Then structures including three soft tissue organs (bladder, bowelbag, small intestine) and five bone organs (left and right femoral head, left and right femur, pelvic) were segmented automatically by deep learning neural network. Performance was evaluated by dice similarity coefficient（ DSC） and Hausdorff distance, using filter back projection(FBP) as the reference. Results Auto-segmentation of deep learning is greatly affected by ASIR-V, but less affected by convolution kernel, especially in soft tissues. Conclusion The stability of auto-segmentation is affected by parameter selection of reconstruction algorithm. In practical application, it is necessary to find a balance between image quality and segmentation quality, or improve segmentation network to enhance the stability of auto-segmentation.
Algorithms ; Humans ; Image Processing, Computer-Assisted ; Neural Networks, Computer ; Radiation Dosage ; Tomography, X-Ray Computed

Objective To explore the urine BLCA-4 test application in the early diagnosis of bladder canc-er.Methods The patient is made up of three groups,62 cases of bladder cancer group,71 cases of bladder be-nign lesions group and 80 cases of healthy physical examination,morning urine specimen collection,ELISA was used to detection BLCA-4 content in morning urine,Statistical analysis differences of urine BLCA-4 ex-pression between three groups,and analysis the relationship between urine BLCA-4 content and the patients with bladder cancer clinical pathological indicators.Results Urine bladder cancer group BLCA-4 levels was higher than the bladder benign lesion group and healthy physical examination group(P<0.05),but no statisti-cal differences between bladder benign lesion group and healthy physical examination group(P>0.05).There is no statistical differences between stage Ta,stage T1-T2 and stage T3-T4,also no statistical differences be-tween G1 and G2-G3;The patients who tumor diameter≥3.0 cm patients urine BLCA-4 levels was higher than tumor diameter < 3.0 cm group(P< 0.05),The sensitivity、specificity and accuracy of diagnosis of urine BLCA-4 test in diagnosis of bladder cancer respectively,95.16%(59/62),97.35%(147/151)and 96.71%(206/213).Conclusion Urine BLCA-4 test has good sensitivity and specificity in diagnosis of bladder cancer, it is very important in bladder cancer screening and monitoring of recurrence of bladder cancer.

Objective: To observe the safety and effects of application of analgesic and sedative drugs in severely burned patients during shock stage. Methods: One hundred and eighty patients with severe burns, conforming to the study criteria, were admitted to our unit from August 2014 to August 2016. Patients were divided into analgesia and sedation group and control group according to whether receiving analgesic and sedative treatment or not, with 90 cases in each group. Patients in control group received conventional treatment, while those in analgesia and sedation group received analgesic and sedative treatment for 24 hours besides conventional treatment. Before and at drug administration hour 2, 8, 16, and 24, pain degree of patients in two groups was scored by visual analogue scale (VAS). At drug administration hour 2, 8, 16, and 24, sedation degree of patients in two groups was scored by richmond agitation sedation scale, and the success rate of sedation was calculated. Mental state of patients within 24 hours of drug administration was observed, while pulse oxygen saturation (SpO₂), respiratory rate, heart rate, and blood pressure were observed and dynamically evaluated every 2 hours. The accidental extubation, tachycardia, hypertension, hypoxia, bradycardia, hypotension, urinary retention, and respiratory depression of patients within 24 hours of drug administration were monitored and recorded. Data were processed with analysis of variance for repeated measurement, one-way analysis of variance, t test, chi-square test, Wilcoxon rank sum test, and Fisher′s exact probability test. Results: (1) The VAS scores of patients in two groups were close before drug administration (t=0.675, P>0.05). The VAS scores of patients in analgesia and sedation group at drug administration hour 2, 8, 16, and 24 were (3.8±0.4), (3.9±0.6), (3.9±0.5), and (3.9±0.9) points, respectively, significantly lower than (6.0±0.9), (6.0±1.2), (6.2±0.6), and (6.3±0.4) points in control group (t=0.785, 0.730, 0.805, 0.895, P<0.05). The success rate of sedation of patients in analgesia and sedation group at drug administration hour 2, 8, 16, and 24 were 91.1% (82/90), 86.7% (78/90), 93.3% (84/90), and 90.0% (81/90), respectively, significantly higher than 7.8% (7/90), 6.7% (6/90), 14.4% (13/90), and 5.6% (5/90) in control group (Z=8.035, 7.946, 8.129, 8.014, P<0.05). (2) The respiratory rate of patients in analgesia and sedation group at drug administration hour 8, 16, and 24 were (15.78±0.69), (16.08±0.59), and (16.21±0.20) times per minute, and the heart rate were (87±9), (83±7), and (76±9) times per minute, respectively, significantly lower than (16.80±0.81), (17.09±0.50), and (17.02±0.61) times per minute and (89±8), (86±7), and (85±6) times per minute in control group (t=7.655, 7.022, 6.536, -6.931, -7.053, -10.196, P<0.01). There were no statistically significant difference in SpO₂, systolic blood pressure, and diastolic blood pressure before and at drug administration hour 2, 8, 16, and 24 between the two groups (t=3.417, -2.894, -6.501, -3.719, -4.573, 2.336, 3.315, 0.942, -1.583, 1.907, 1.147, -0.968, 0.931, -1.682, 1.076, P>0.05). (3) The rates of respiratory depression, hypoxia, bradycardia, urinary retention, and hypotension of patients in the two groups were close (χ²=0.310, P>0.05). The rates of hypertension, accidental extubation, and tachycardia of patients in analgesia and sedation group were significantly lower than those in control group (χ²=16.364, 5.143, 73.309, P<0.05 or P<0.01). Conclusions Proper application of analgesic and sedative drugs in severely burned patients during shock stage has good clinical effect with low incidence rates of complications.

Objective By analysis of the key performance indexes of the clinical laboratory automation system, to clarify the advantage and optimize the comprehensive performance of the laboratory automation system.Methods Key performance indexes were Collected from January 2017 to April 2017 in biochemistry and immunoassay group of Clinical Laboratory of Shanghai Tong Ren Hospital.(1)The data were collected and compared by the before-and-after method,the starting time of the automation system and initial sample test were analyzed.(2)Key performance indexes were analyzed for the time of specimen registration,inspection,and reporting.(3)The specimen turnaround time(TAT)was analyzed based on two months operation of the laboratory automation system.In view of disadvantage of infectious assays, setting up priority sample absorption, then TAT performance was re-evaluated.(4)By the assessment of total serum dosage required in the automation system, the number of blood vacuum tubes were reduced reasonably.The pros and cons of laboratory automation system were analyzed and the potential improvement were proposed.Results (1)According to the sample peak shift forward,the system start time could move forward 30 minutes earlier.(2)With the adopting of railway logistics,the specimens were sent to the lab and the registration time was at 7:25 am,and the time required for specimen delivery was greatly reduced which made specimen test,report and audit time all moved forward accordingly.(3)Data has shown that specimen TAT declined dramatically based on the performance of the first two month operation of the automation system,biochemical items were shortened 2 h,and the immunoassay shortened 4 h,respectively.Moreover the trend keeps better gradually.With setting up priority absorption infectious tests,the TAT was improved greatly,TAT reduced the average by 40 min.(4)500 μl(including the sample in dead space of vacuum tube)were needed for all the 65 biochemical items included in the system, and 1 495 μl serum were used for the 28 immunoassay.As a result, a total of 2 000 μl serum will be enough for sample analysis by the system, which provided the feasibility to reduce 3 vacuum tubes averagely.Considering the current automation system does not include all the analysis items in our lab directory, a few tests remain to be performed on offline instruments respectively.The methodology for some infectious agents are different from previous method, therefore some test results may need a period of time for comprehensive clinical appreciation.Furthermore,due to the parallel connection of multiple instruments included in the system, more rigorous and frequent quality control becomes a necessity,which may rely on more strict quality control procedure to guarantee the quality.Conclusions The application of the automation system significantly enhanced the efficiency of clinical laboratory all round.In addition, by the quantitative indicators, it is possible to monitor the system operation performance real time, which may feedback and facilitate the improvement constantly,and result in auto confirmation the majority results,eventually.

[Abstract ] Objective To understand the identification and in vitro antifungal susceptibility of Penicillium marneffei(PM)in yeast phase,and guide clinic antifungal application.Methods Strains isolated from blood and bone marrow of 23 patients infected with PM in a hospital between 2009 and 2016 were collected,colony morpholo-gy of PM in yeast phase was observed,susceptibility to itraconazole,voriconazole,amphotericin B,and fluconazole were detected with E-test method.Results Colony morphology of PM were as follows:direct microscopic examina-tion of Wright’s staining of tissue specimens found visible oval or round spore with apparent septum,and mainly lo-cated in macrophage;Gram staining of blood culture specimens found that strains were with bulbous and slightly curved ends,occasionally branched and with septum. PM was dimorphic fungi,presented mycelium at 28° C,pro-duced red pigment and diffused into medium;PM presented yeast form at 35° C,there were typical colony morpholo-gy. Minimum inhibitory concentrations (MICs)of itraconazole,voriconazole,amphotericin B,and fluconazole to PM in yeast phase were 0.002-0.016,0.012-0.125,0.002-0.500,and 0.500-16.000μg/mL respectively. Conclusion Typical colony morphology and fungal spore of PM in bone marrow and peripheral blood are important features for identification. PM is most susceptible to itraconazole,followed by voriconazole and amphotericin B, while fluconazole is less susceptible.

Objective:To study the effect of alum on immune response in mice induced by HBoV1 VP2 VLPs.Methods:BABL/c mice were randomly divided into VLPs experimental group, alum adjuvant experimental group, PBS control group and alum adjuvant control group,the experimental group mice were intramuscular immunization with HBoV1 VP2 VLPs and HBoV1 VP2 VLPs added alum,control group mice were immunization with alum or PBS buffer,then to study the effect of alum on immune response in mice induced by HBoV1 VP2 VLPs by cellular and humoral immune strength.Results: Alum adjuvant decreased cellular immune response induced by HBoV1 VP2 VLPs(P<0.001),enhance the HBoV1 VP2 VLPs immuned serum IgG titer(P<0.05)and IgG activity(P<0.01).Conclusion:Alum adjuvant can enhance humoral immune response induced by HBoV1 VP2 VLPs,but weaken cellular immune response induced by HBoV1 VP2 VLPs,when HBoV1 VP2 VLPs used as a prophylactic vaccine it should add alum adjuvant,when used as a therapeutic vaccine,it should not add alum adjuvant.

Objective To investigate the value of transvaginal ultrasound for the diagnosis of endometrial polyps .Methods The ultrasonographic characteristics in 56 cases of endometrial polyp diagnosed by transvaginal ultrasound were performed the retro-spective analysis and the situation of the hysteroscopic examination in consistent with the pathological examination was analyzed . Results Among 56 cases ,50 cases of endometrial polyps were verified by the pathological examination ,the coincidence rate of transvaginal ultrasound diagnosis was 89 .3% ,in which 6 cases were misdiagnosed with the misdiagnosis rate was 10 .7% . Conclusion The transvaginal ultrasound has the high accuracy rate for diagnosing endometrial polyps .

Objective To evaluatc the efficacy and safcty of recombinant endostatin (Endostar)combined with concurrent radio-chemotherapy (CRCT) in patients with unresectable stage Ⅲ non-small cell lung cancer (NSCLC).Methods From March 2009 to November 2011,47 patients received threedimensional conformal radiotherapy of 60-66 Gy in 30-33 fractions over 6-7 weeks And concurrent chemotherapy of docetaxel 65 mg/m2 and cisplatin 65 mg/m2.Endostar was administered once a week before and on week 2,4,6 during CRCT at a dose level of 7.5 mg/m2/d.Tumor response was evaluated with thoracic CT scans performed 4 weeks after completion of treatment in accordance with RECIST 1.1 criteria.Acute toxicities were evaluated in accordance with CTCAE 3.0.Results Forty-four patients completed treatment and toxicity evaluation,42 patients completed evaluation of efficacy.Five patients achieved complete response,29 partial response,3 stable disease,and 5 progressive disease,2 were net assessed.Overall response rate was 77％.One-year overall survival rate was 81％,and one-year progression-free survival rate was 51％.Twelve patients died,2 died of treatment related toxicities,8 of cancer,and 2 of unknown causes.Nineteen patients developed grade 3/4 neutrocytopenia,grade 3 acute esophagitis and pneumonitis were observed in 4 and 4 patients,respectively,and 1 patient died of pneumonitis.No patient developed cardiovascular toxicities and hemorrhage.Conclusions Endostar combined with CRCT for unresectable stage Ⅲ NSCLC was safe and the short term outcomes were promising.Further investigations are warranted.

Objective To access the effects of aldosterone (ALD) and its receptor antagonist- spironolactone (SPI) on the production of reactive oxygen species (ROS) and apoptosis in podocytes and to explore the possible mechanism involved. Methods Conditionally immortalized mouse podocytes were divided into control group, ALD group, SPI group and SPI +ALD group. The level of ROS production and the expression of nephrin protein were assayed by fluorescent spectrophotometry and indirect immunofluorescence technology. The apoptosis rate of podocytes was monitored by flow cytometry. The expression of bax and bcl-2 mRNA and protein was detected by RT-PCR and Western blotting methods. The anti-oxidant N-acetylcysteine (NAC)was applied to determine whether the effects of ALD on podocytes were mediated by ROS pathway.Results Compared with the control group, ALD significantly increased ROS production in podocytes (P＜0.05). SPI completely abolished the above-mentioned effect of ALD (P＜0.05). ALD induced the down-regulation of the expression of nephrin and the up-regulation of podocytes apoptosis (P＜0.05), which was accompanied with the elevated expression of bax mRNA and protein and the reduced expression of bcl-2 mRNA and protein (P＜0.05). SPI or NAC prevented the above-mentioned changes induced by ALD (P＜0.05). Conclusion ALD increases theexpression of pro-apoptotic factor (bax) but decreases the expression of anti-apoptotic factor (bcl-2)to induce podocytes apoptosis through the mineralocorticoid receptor (MR) possibly via the mechanisms involving ROS pathway.