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 To investigate the population distribution of non-alcoholic fatty liver disease before and after renaming and the association between the types of metabolic risk factors(MRF)for metabolic dysfunction-associated steatotic liver disease(MASLD)and advanced liver fibrosis.Methods This study was conducted among 515 patients who were admitted to The Affiliated Hospital of Xuzhou Medical University and Wuxi Fifth People's Hospital from January 2019 to January 2022 and had hepatocyte steatosis≥5%by liver biopsy.Among these patients,2 patients did not meet the diagnostic criteria for nonalcoholic fatty liver disease(NAFLD)and metabolic associated fatty liver disease(MAFLD),respectively,and were classified as steatotic liver disease(SLD)with other specific causes,and the other 513 patients were divided into MASLD group with 275 patients,comorbid group with 216 patients(MASLD comorbid with other liver diseases),and cryptogenic SLD group with 22 patients.The above groups were compared in terms of clinical features,laboratory markers,and advanced liver fibrosis.The MASLD patients with different types of MRF were compared in terms of clinical features,laboratory markers,and advanced liver fibrosis,and the risk factors for advanced liver fibrosis in patients with MASLD were analyzed.The Kruskal-Wallis H test was used for comparison of continuous data with skewed distribution between multiple groups and further comparison between two groups;the chi-square test was used for comparison of categorical data between multiple groups,and Bonferroni correction was used for further comparison between two groups.The logistic regression analysis was used to identify the risk factors for liver fibrosis.Results Among the 515 patients with SLD,297 patients(57.7%)met the diagnostic criteria for NAFLD,among whom 22 were classified as cryptogenic SLD and 275 met the diagnostic criteria for MASLD,and 467(90.7%)were diagnosed with MAFLD.There were significant differences between the three groups in sex,body mass index(BMI),gamma-glutamyl transpeptidase,triglyceride,cholesterol,low-density lipoprotein cholesterol,high-density lipoprotein cholesterol,fasting plasma glucose,NAFLD fibrosis score(NFS),fibrosis-4(FIB-4),and F3-4(all P＜0.05).Compared with the MASLD group and the cryptogenic SLD group,the comorbid group had the highest proportion of patients with advanced liver fibrosis(P＜0.001).With the increase in the type of MRF,the patients tended to have an older age,a significantly higher proportion of female patients,a higher possibility of hypertension and diabetes,and higher levels of metabolic parameters including BMI,blood lipids,and blood glucose(all P＜0.05).With the increase in the types of MRF in MASLD patients,they tended to have significantly higher noninvasive fibrosis scores(NFS and FIB-4)and a significantly higher proportion of patients with advanced liver fibrosis(P＜0.05).The multivariate logistic regression analysis showed that age≥50 years(odds ratio[OR]=2.622,95%confidence interval[CI]:1.091-6.300,P=0.031)and the increase in the type of MRF(OR=1.876,95%CI:1.194-2.947,P=0.006)were independent risk factors for MASLD with severe liver fibrosis.Conclusion The new definition of MASLD is based on the positive identification of MRF,and the reclassified population of MASLD is smaller than that of MAFLD,with little difference from that of NAFLD.In addition,age≥50 years and the increase in the type of MRF are independent risk factors for MASLD with advanced liver fibrosis.

Objective:To evaluate the safety of naborphine hydrochloride combined with propofol in painless colonoscopy diagnosis and treatment of hypertensive patients.Methods:From October 2018 to September 2020, 900 patients with ASA grade Ⅰ to Ⅲ, aged 18 to 65, who underwent colonoscopy in Zhuhai Hospital Affiliated to Jinan University and Shanghai East Hospital Affiliated to Tongji University were prospectively selected. According to the random number table method, the patients were divided into 3 groups ( n=300): naborphine hydrochloride group 1 (N1 group, intravenous injection of 0.05 mg/kg naborphine hydrochloride); naborphine hydrochloride group 2 (N2 group, intravenous injection of 0.1 mg/kg naborphine hydrochloride); sufentanil group (SF group, intravenous injection of 0.1 μg/kg sufentanil). During anesthesia induction, propofol was combined with sedation, and the dose of propofol was 1.5 mg/kg. The systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), pulse oxygen saturation (SpO 2), respiratory rate (RR), and mean arterial pressure (MAP) of the three groups were compared before anesthesia (T 0), during induction (T 1), 1 min after induction (T 2), 2 min after induction (T 3), 3 min after induction (T 4) and 4 min after induction (T 5), and the bispectral index (BIS) were monitored. At the same time, the examination time, total dosage of propofol, recovery time, postoperative Visual Analogue Scale (VAS) score and perioperative anesthesia related adverse reactions of the three groups were compared. Results:There was no significant difference among the three groups in examination time, total dosage of propofol, recovery time, postoperative VAS score and adverse anesthetic reactions (all P>0.05). There was no significant difference in HR, SpO 2 and RR among the three groups at different time points (all P>0.05). The SBP, DBP and MAP in N1 group at T 1, T 3, T 4 and T 5 were lower than those in SF group (all P<0.05); The SBP, DBP and MAP in N2 group at T 1, T 3 and T 4 were higher than those in N1 group (all P<0.05). The BIS in T 3 and T 4 of N2 group was higher than that of N1 group (all P<0.05). Conclusions:0.1 mg/kg naborphine hydrochloride combined with propofol for painless enteroscopy in patients with hypertension has fine anesthetic effect and safety.

Objective To investigate the CT findings of primary orbital lipid prolapse. Methods The CT data of 13 patients with primary orbital lipid prolapse in Jiaxing Traditional Chinese Medicine Hospital Affiliated to Zhejiang Chinese Medical University from January 2012 to June 2018 were retrospectively analyzed. Results In all of the 13 cases of primary orbital lipid prolapse, there were 12 cases of bilateral orbital lipid prolapse and 1 case of unilateral orbital lipid prolapse. The CT images showed that the fat density masses in the posterior orbital septum were herniated from the outer quadrant of the orbit. The tumor was crescent shaped and connected with the orbital adipose tissue. The mass and the orbital adipose tissue were natural and homogeneous. Conclusions CT has distinctive imaging findings in the diagnosis of primary orbital lipid prolapse, and it is helpful for the diagnosis and differential diagnosis of this disease.

Objective: To analyze the mid-term clinical efficacy of extended to C1, 2 posterior single-door laminoplasty for the treatment of ossification of the posterior longitudinal ligament combined the upper cervical spine. Methods: From February 2013 to December 2015, 32 patients with cervical OPLL who were extended to C1, 2 posterior open-door laminoplasty were enrolled in our hospital (complete follow-up of 25 patients). There were 19 males and 6 females, aged 48-76 years (mean 60.1±7.0 years). Cervical spinal cord function was assessed using the Japanese Orthopaedic Association (JOA) spinal cord function score, and the rate of improvement in neurological function was calculated. The quality of life of patients was assessed using the neck disability index (NDI). Axial symptoms were assessed and indexed using the visual analog scale (VAS). The C0-2 angle of the cervical spine was measured by X-ray preoperative and at the last follow-up. The cervical curvature was evaluated by the C2-7 angle, and the cervical vertebra activity was observed by the dynamic X-ray of the cervical spine. Results: 25 patients were followed up for a period of 26-64 months (mean 35.9±8.1 months). At the last follow-up, the JOA score was 14.32±3.24, the NDI score was 7.61±1.23, and the VAS score was 1.42±0.78. The differences between the three scores were statistically significant compared with the preoperative scores. The neurological improvement rate at the last follow-up was 79.61%±13.23%. The preoperative C0-2 angle was 26.04°±6.28°, and the last follow-up was 24.92°±5.51°; the C2-7 angle was 19.55°±9.42° before surgery, and the C2-7 angle at last follow-up was 17.97°±8.80°. The C2-7 angle at last follow-up was slightly reduced compared with that preoperative, but the difference was not statistically significant. The preoperative cervical vertebra ROM was 35.31°±12.24°, and at the last follow-up it was 32.23°±9.65°. The ROM of cervical vertebrae at the last follow-up was slightly lower than that before surgery, but the difference was not statistically significant. Among them, the reduction of the ROM of overflexion was greater than that of the over-extension, which was the main reason for the decrease of ROM. The OPLL bone mass continued to grow at the last follow-up of 11 patients. Conclusion The extended to C1, 2 posterior single-door laminoplasty for the treatment of ossification of the posterior longitudinal ligament combined the upper cervical spine can achieve adequate spinal cord decompression, satisfactory neurological improvement, and improve the postoperative cervical curvature and activity. There was no obvious change in the curvature and activity of the cervical spine, and the axial symptoms did not increase significantly, and the clinical efficacy was positive.

OBJECTIVE: To verify whether dexmedetomidine hydrochloride (Dex) alleviates renal ischemia-reperfusion (IR) injury in diabetic rats by increasing the expression of hypoxia inducible factor-1 (HIF-1). METHODS: A rat model of type 2 diabetes mellitus was established by high-fat diet and streptozotocin injection. The rats were subjected to daily intragastric administration of 0.05 mg/kg digoxin for 7 consecutive days and intraperitoneal injection of Dex 2 h before renal IR injury induced by ligation of the bilateral renal arteries for 60 min followed by reperfusion for 120 min. After reperfusion, blood samples were taken for detection of serum creatinine (Scr) and urea nitrogen (BUN) levels. Western blotting was used to detect the expression of HIF-1, cleaved caspase-3, Bcl-2, and Bax in the renal tissues; the expression of the HIF-1, p-eNOS, and eNOS were detected using ELISA. The percentage of apoptotic glomerular cells was assessed using TUNEL assay. RESULTS: The levels of Scr, BUN, HIF-1, p-eNOS, and eNOS and the percentage of apoptotic cells in both normal and diabetic rats increased significantly after renal IR injury ( < 0.05). The expressions of Scr, BUN, p-eNOS, and eNOS decreased while HIF-1 expression increased significantly in Dex-treated rats with renal IR injury ( < 0.05). Compared with the non-diabetic rats, the diabetic rats showed more obvious increase in the expressions of Scr, BUN, p-eNOS, and eNOS following renal IR injury. In the diabetic rats with renal IR injury, Dex treatment prior to the injury significantly lowered the expressions of Scr, BUN, p-eNOS, eNOS, cleaved caspase-3, and Bax, decreased the percentage of apoptotic cells, and increased the levels of HIF-1a and Bcl-2 ( < 0.05). Digoxin treatment significantly antagonized the effects of Dex in the diabetic rats with renal IR injury by increasing the expressions of cleaved caspase-3 and Bax, promoting glomerular cell apoptosis, and decreasing renal expressions of HIF-1 and Bcl-2 ( < 0.05). CONCLUSIONS Dex alleviates renal IR injury in diabetic rats probably by inhibiting renal expression of HIF-1 and glomerular cell apoptosis.
Animals ; Dexmedetomidine ; Diabetes Mellitus, Experimental ; Diabetes Mellitus, Type 2 ; Hypoxia ; Hypoxia-Inducible Factor 1, alpha Subunit ; Kidney ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury

OBJECTIVE: To analyze the relationship between and gene methylation with aging in the general population. METHODS: We collected peripheral blood samples from 284 male and 246 female healthy subjects for detection of methylation levels of and genes using quantitative methylation-specific PCR (qMSP). The relationship between the methylation levels of and genes and aging was analyzed using Spearman or Pearson correlation test. RESULTS: We found a significant positive correlation between the methylation levels of the two genes in these subjects ( < 0.05). In the overall population as well in the female subjects, methylation was found to be inversely correlated with age ( < 0.05). The methylation levels of and genes were inversely correlated with TG, ApoE, Lp(a) and AST in the overall population ( < 0.05). In both the female and male subjects, the methylation levels of the two genes were inversely correlated with Lp(a) ( < 0.05). In the male subjects, methylation was inversely correlated with AST ( < 0.05), while methylation was inversely correlated with HDL and ApoE ( < 0.05). In the female subjects, methylation was positively correlated with LDL and inversely correlated with ApoE and AST ( < 0.05). CONCLUSIONS The methylation levels of and are closely related to age and the levels of multiple proteins in healthy subjects.
Aging ; Cyclin-Dependent Kinase Inhibitor p15 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; DNA Methylation ; Female ; Humans ; Male ; Real-Time Polymerase Chain Reaction

This paper systematically summarized the core problems in the field of medical and health services and did the root analysis.It is concluded that the current domestic and international related medical service modes deviated from the essence of health services and led to difficulties in the idea and method,bringing people to new problems and new crises.KY3H health care service model,set innovative ideas,innovative technical methods,innovative products,innovative processes,standards and standards into one,by providing a full range and the whole cycle of people-oriented,self-help,"diagnosis and treatment of security" integration of health careservice,improve people's health status,fundamentally change the traditional passive "seeking medical treatment" service mode,to active "seeking one's own" health service mode,so as to achieve "health status continues to rise,the medical costs continue to decline,"which is the social development aims.The paper systematically analyzes the idea,operational logic,goal,mode structure,characteristics of the model,and its significance to individuals,industries,society,countries and even the international community in order to make a positive contribution to the healthy development strategy in China and provide a Chinese case for global Health.

Vigorously development of the health service industry has received great attention,but the health service industry faces many difficulties.By analyzing the current predicament,this paper put forward the KY3H health care service model and systematically analyzed its six innovations:from "disease as the center" to "health as the center",from "homogenized group health service" to "Personalized and Humanized Health Service",from "Extensive Health Service"to "Precise Intervention of Individual Health Status",from "Single Health Service" to "Trinity Service of Health,Culture,Health Management and Health Insurance",from the "seeking medical" service model to "seeking one's own"service model,as well as from the "fixed (fixed-time,fixed-point,fixed-mode) service" to "anytime,anywhere full-time service".This model has achieved initial success in practice.It is an important way to solve the dilemma.

Objective To evaluated the relationship of ambulatory pulse pressure and severity intracranial arterial stenosis. Methods 165 ischemic stroke patients with intracranial arterial stenosis were selected as stenosis group. And 142 patients with no obvious intracranial arterial stenosis were selected as control group. All patients were given ambulatory blood pressure monitoring. 24 h average systolic pressure ,diastolic pressure and ambulatory pulse pressure were measured. Results Ambulatory pulse pressure of stenosis group[(73.02 ± 10.86)mmHg]was significantly higher than that of control group[(59.63±12.96)mmHg,P<0.01]. Ambulatory pulse pressure of mod-erated stenosis group[(69.73±9.3)mmHg,P < 0.05]was significantly higher than that of control group[(59.63± 12.96)mmHg,P<0.01]. Ambulatory pulse pressure of severe stenosis group[(77.36±7.94)mmHg]was signifi-cantly higher than that of moderate stenosis group[(69.73±9.3)mmHg,P < 0.05]. The level of ambulatory pulse pressure was independent risk factors of intracranial arterial stenosis(OR value:1.092). Conclusions Ambulatory pulse pressure is significantly related to the severity of intracranial arterial stenosis and is an independent risk factor for intracranial arterial stenosis.