Lycium barbarum polysaccharide inhibits starvation-induced apoptosis of Schwann cells by enhancing autophagy / 神经解剖学杂志

Objective:To investigate the inhibitory effect of lycium barbarum polysaccharide(LBP)on apoptosis of Schwann cells(SCs)and its related mechanisms.Methods:The autophagy model was prepared by starvation treatment of RSC96 cells for 12 h,and the expressions of autophagy related proteins LC3 and p62 were detected by Western Blot.Cell Counting Kit-8(CCK-8)kits were used to detect the optimal concentration of LBP.RSC96 cells were randomly divided into Control group,Starvation group and Starvation+LBP group.The expressions of autophagy associated pro-teins(LC3,p62)and myelin associated proteins(p75NTR,PMP22,S100β)were detected by Western Blot or immu-nofluorescence staining.Annexin V/PI fluorescence staining was used to detect apoptosis of the cells.The cell cycle was analyzed by flow cytometry.Western Blot analysis of phosphorylation levels of pathway proteins Erk1/2 and Akt.Results:CCK8 results showed that the viability of damaged RSC96 cells was the best when LBP was 300 μg/ml.Com-pared with Control group,LC3-Ⅱ/LC3-I levels in Starvation group were significantly increased(P＜0.05).Compared with Starvation group,the proportion of apoptotic and necrotic cells in Starvation+LBP group was significantly de-creased,and the proportion of cells in S and G2/M stages was increased.The expression levels of LC3-Ⅱ,p75NTR,PMP22 and S100β were increased,while the expression levels of autophagy substrate protein p62 were decreased.In-creased expression of pathway protein p-Erk1/2(P＜0.05),while the expression of p-Akt protein decreased slightly.Conclusion:LBP can inhibit the apoptosis of SCs and promote the expression of myelin-related proteins by enhancing autophagy,which is related to the activation of Erk1/2 and/or the inhibition of Akt.

Study on enzymatic hydrolysis conditions of cannabis,MDMB-4en-PINACA and ADB-BUTINACA in urine / 中国法医学杂志

Objective Synthetic cannabinoids(SCs)will be widely metabolized to phase I metabolites and glucuronide metabolites after entering human body.It is usually difficult to detect the parent drug of SCs in urine.Therefore,it is necessary to undergo de-glucuronidation in the preparation of urine samples.We aimed to optimize the enzymatic hydrolysis methods for SCs detection in urine.Methods We studied enzymatic hydrolysis for the determination of 11-demethyl-9-carboxyl-THC(Δ9-THC-COOH)in THC positive urine samples by liquid chromatography-tandem mass spectrometry(LC-MS/MS),and compared it with the alkaline digestion method.Meanwhile,we studied enzymatic hydrolysis for the determination of the related metabolites in MDMB-4en-PINACA and ADB-BUTINACA positive urine samples.Results The Δ9-THC-COOH glucuronic acid conjugate could be hydrolyzed by adding 3μL β-d-glucuronidase solution(＞100 000 units/mL)for 30 min at 55℃.The MDMB-4en-PINACA M and ADB-BUTINACA M glucuronide conjugates were hydrolyzed by adding 3μL β-d-glucuronidase solution(＞100 000 units/mL)for 30 min at 75℃.Conclusion This study can provide reference for the establishment of rapid,accurate and reliable enzymatic hydrolysis methods when detecting SCs in urine.

Estimation of Runoff Using USDA SCS-Curve Number and Autoregressive Time Series Model Parameters for Kachhinda Watershed, Morena District, Madhya Pradesh, India

Hydrological modeling is an influential method for examining hydrologic systems, serving as a valuable tool in investigating these systems for both present studies involving two hydrologic runoff models viz. Soil Conservation Service Two methods, namely the Soil Conservation Service-Curve Number (SCS-CN) method and the Autoregressive Time Series model, were utilized in the Kachhinda watershed to estimate surface runoff, CN, and AMC conditions. The study area covers 600 hectares and is sited in Morena district of M.P. in Chambal division. The SCS-CN technique was employed to determine the curve number and estimate surface runoff by using the potential maximum retention. The CN of the watershed was calculated and compared to observed and estimated surface runoff, which were found to be in close agreement with each other. Additionally, an Autoregressive Time Series model was developed to establish the correlation between observed and estimated runoff, resulting in a correlation coefficient of 0.974. Different orders of AR time series models (0, 1, and 2) were tested to predict annual stream flow, and the model goodness of fit was evaluated using the Box-Pierce Portmanteau test and the Akaike Informations Principle. The Akaike Information Criterions value for the Autoregressive (1) model for runoff was found to be 0.919158, which is within the range of the values obtained for Autoregressive (0) (0.207433) and Autoregressive (2) (5.9767) models.

Development status and new challenges of heart transplantation / 器官移植

Heart transplantation is the primary therapeutic option for patients with end-stage heart failure. The shortage of donors has been the main limiting factor for the increasing quantity of heart transplantation. With persistent updating and introduction of novel technologies, the donor pool has been increasingly expanded, such as using the heart from older donors, donors infected with hepatitis C virus, donors dying from drug overdose or donation after cardiac death (DCD) donors, etc. Meantime, the proportion of recipients with advanced age, multiple organ dysfunction, mechanical circulatory support and human leukocyte antigen antibody sensitization has been significantly increased in recent years. The shortage of donors, complication of recipients' conditions, individualized management of immunosuppressive therapy and prevention and treatment of long-term cardiac allograft vasculopathy are all challenges in the field of heart transplantation. In this article, novel progresses on donor pool expansion, improving the quality of recipients, strengthening the diagnosis and treatment of rejection, and preventing cardiac allograft vasculopathy were reviewed, aiming to prolong the survival and enhance the quality of life of patients with end-stage heart failure on the waiting list or underwent heart transplantation.

Normothermic machine perfusion of kidney and ischemia-free kidney transplantation / 器官移植

To increase the utilization rate of expanded criteria donor (ECD) kidney, the kidney preservation methods have been ever advancing in recent years. The application of normothermic machine perfusion (NMP) promotes the preservation, evaluation and repair of ex vivo donor kidneys and accelerates the innovation of surgical approaches of kidney transplantation. Ischemia-free kidney transplantation (IFKT), which initiated by Organ Transplantation Center of the First Affiliated Hospital of Sun Yat-sen University, keeps the blood flow and oxygen supply of the donor kidney with NMP machine during the entire process of acquisition, preservation and transplantation, thereby fundamentally avoiding ischemia-reperfusion injury (IRI) of the donor kidney and reducing the risk of delayed graft function (DGF) and acute rejection after surgery. In this article, recent progresses upon the kidney NMP, surgical procedures and short-term outcomes of IFKT were reviewed, aiming to provide reference for enhancing the utilization rate of ECD donor kidney and resolving the issue of organ shortage.

Involvement of Opioid Peptides in the Analgesic Effect of Spinal Cord Stimulation in a Rat Model of Neuropathic Pain / 神经科学通报·英文版

Spinal cord stimulation (SCS)-induced analgesia was characterized, and its underlying mechanisms were examined in a spared nerve injury model of neuropathic pain in rats. The analgesic effect of SCS with moderate mechanical hypersensitivity was increased with increasing stimulation intensity between the 20% and 80% motor thresholds. Various frequencies (2, 15, 50, 100, 10000 Hz, and 2/100 Hz dense-dispersed) of SCS were similarly effective. SCS-induced analgesia was maintained without tolerance within 24 h of continuous stimulation. SCS at 2 Hz significantly increased methionine enkephalin content in the cerebrospinal fluid. The analgesic effect of 2 Hz was abolished by μ or κ opioid receptor antagonist. The effect of 100 Hz was prevented by a κ antagonist, and that of 10 kHz was blocked by any of the μ, δ, or κ receptor antagonists, suggesting that the analgesic effect of SCS at different frequencies is mediated by different endorphins and opioid receptors.

Research progress on application of machine perfusion in quality maintenance of donor kidney from organ donation after citizen's death / 器官移植

Organ donation after citizen's death has become the main source of organ donation in China. However, the complexity of donor quality and the increasing proportion of expanded criteria donor (ECD) exert significant impact upon the availability of donor kidney and the long-term prognosis of recipients after kidney transplantation. Strengthening the quality maintenance and evaluation of donor kidney is of great significance for improving the quality of donor kidney, increasing the procurement and utilization of donor kidney and prolonging the long-term survival of recipients and kidney allografts. As one of the major approaches of organ preservation, machine perfusion preservation may not only prolong the preservation time and improve the quality of donor kidney, but also play a critical role in the repair and function evaluation of donor kidney. Based on literature review, several hot issues, corresponding treatment strategies and research progress on machine perfusion in the quality maintenance of donor kidney from organ donation after citizen's death were reviewed in this article, aiming to provide reference for selecting the optimal preservation method of donor kidney and enhancing the quality and utilization rate of ECD donor kidney.

miRNA-223 expression in patient-derived eutopic and ectopic endometrial stromal cells and its effect on epithelial-to-mesenchymal transition in endometriosis

Abstract Objective: This study was designed to evaluate the expression of microRNA-223 (miRNA-223) in patient-derived eutopic and ectopic endometrial stromal cells (SCs). Given the fact that miRNA-223 was previously shown to be upregulated in these cells and that this upregulation has been linked to epithelial-to-mesenchymal transition (EMT) during endometriosis, this study aimed to further explore the expression of miRNA-223, its effect in endometriosis, and the mechanisms underlying its effects. Methods: Endometrial tissue was collected from 26 patients with endometriosis and 14 patients with hysteromyoma (control group). Primary endometrial SCs were isolated and cultured from several endometrial samples and miRNA-223 expression was evaluated using qRT-PCR. Cells were then transfected with a miRNA-223 overexpression lentiviral vector (sh-miR-223 cells) or an empty control (sh-NC cells) and then used to monitor the effects of miRNA-223 on the expression of several EMT-associated proteins, including N-cadherin, vimentin, and Slug, using western blot. Cellular migration, invasion, and proliferation were then evaluated using a wound healing, Transwell, and CCK-8 assay, respectively. Flow cytometry was used to detect apoptosis. Results: There was a significant decrease in the expression of miRNA-223 in both eutopic and ectopic endometrial SCs (p < 0.05) whereas upregulation of miRNA-223 inhibited the expression of EMT-related molecules and reduced cell migration, invasion, and proliferation. High levels of miRNA-223 also promoted apoptosis. Conclusion: miRNA-223 expression decreased in endometrial SCs from endometriosis patients, which may facilitate the differential regulation of EMT during endometriosis. Clinical Trial registration number: SWYX2020-211.

Clinical application of biomarkers in DCD donor kidney perfusate for predicting delayed graft function after renal transplantation / 器官移植

Objective To explore the feasibility of biomarkers in static cold storage (SCS) perfusate of donor kidney from donation after cardiac death (DCD) for predicting delayed graft function (DGF) after renal transplantation. Methods Clinical data of 64 recipients and 47 donors undergoing DCD renal transplantation were retrospectively analyzed. All recipients were divided into the DGF group (n=7) and immediate graft function (IGF) group (n=57) according to the incidence of postoperative DGF in the recipients. The levels of neutrophil gelatinase-associated lipocalin (NGAL), liver-type fatty acid-binding protein (L-FABP), interleukin -18(IL-18) and kidney injury molecule-1 (KIM-1) in the SCS perfusate were statistically compared between two groups, and the correlation with DGF was analyzed. The predictive value of each biomarker in the occurrence of DGF in recipients after renal transplantation was analyzed. Results The incidence of DGF in the recipients undergoing DCD renal transplantation was 11% (7/64). The NGAL level in the donor kidney perfusate of the DGF group was significantly higher than that in the IGF group (P=0.009). The NGAL level in the donor kidney perfusate was positively correlated with the incidence of DGF in recipients after renal transplantation (r=0.430, P < 0.001). The receiver operating characteristic (ROC) curve analysis showed that the increased levels of NGAL and KIM-1 in the perfusate yielded certain predictive value for DGF in recipients after renal transplantation (both P < 0.05). The area under the curve (AUC) of combined detection of NGAL and KIM-1 for predicting DGF in recipients after renal transplantation was 0.932 [95% confidence interval (CI) 0.850-1.000]. The sensitivity was calculated as 1.000 and 0.754 for the specificity (P < 0.05). Conclusions The NGAL level in the SCS perfusate of DCD donor kidney is associated with the occurrence of DGF in recipients after renal transplantation. Combined detection of NGAL and KIM-1 levels in the perfusate may accurately predict the occurrence of DGF in recipients after renal transplantation.

Development status and clinical applications of precision detection technology in the times of precision oncology / 中国肿瘤生物治疗杂志

@#Precision detection techniques have promoted the development of individualized diagnosis and treatment of tumors in the era of precision medicine. At the same time, clinical demands of precision treatments have further driven the development and application of precision detection techniques. In recent years, precision medical detection techniques realized rapid transformations from low-throughput to high-throughput genomic sequencing, from tissue biopsies to liquid biopsies, and from multicell promiscuous detection to single cell precision sequencing. All these changes have promoted the emergence and development of new technologies, new targets, and new drugs in the era of precision oncology medicine. In the future, multi-dimensional combined detection could help to improve the accuracy of precision medicine; ctDNA methylation detection analysis could broaden the research field of precision medicine; and the transformation of clinical trial design could also contribute to promote the in-depth development of precision medicine.

Microorganismos aislados en cultivo bacteriológico de muestras de leche de vacas holstein clínicamente sanas / Microorganisms isolated in bacteriological culture from milk samples from clinically healthy holstein cows / Microrganismos isolados em cultivo bacteriológico de amostras do leite de vacas holandesas clinicamente saudáveis

Resumen El objetivo del presente estudio fue determinar la frecuencia de aislamiento de microorganismos de tipo infeccioso y ambiental en leche de un grupo vacas clínicamente sanas en dos diferentes tipos de ordeño. Para ello se utilizaron muestras de leche de 289 vacas ubicadas en hatos de cuatro municipios del Norte del departamento de Antioquia en diferentes sistemas de ordeño (manual y mecánico), a cada muestra de leche se le hizo cultivo bacteriológico para determinar la presencia de microorganismos. De 289 muestras de leche evaluadas, 193 (66,78%) fueron positivas para aislamiento de cualquier tipo de patógeno (ocho diferentes patógenosaislados), de las cuales 81 (28,02%) muestras provenían de ordeño manual y 112 (38,75%) pertenecían a ordeño mecánico, encontrando un mayor porcentaje de aislamiento de patógenos bacterianos en muestras de leche provenientes de sistema de ordeño mecánico (p=0,0236). El patógeno de mayor aislamiento fue el Arcanobacterium haemolyticum (antes clasificado como Corinebacterium sp), microorganismo que hace parte de la flora saprófita del animal, con presencia de manera individual en el 20,14% de las muestras analizadas y en confección con otros patógenos en 0,7% de las muestras. El microorganismo que siempre se reporta como mayor productor de mastitis subclínica en bovinos es el Streptococcus agalactiae, un patógeno infeccioso, que en el presente estudio se aisló del 12,50% de las muestras de leche. Se determinó el Odds ratio (OR), entre aislamiento de Streptococcus agalactiae y Score de Células somáticas (SCS) que fue estadísticamente significativo, indicando que cuando está presente este patógeno el SCS aumenta y los animales son más susceptibles a padecer mastitis.

Abstract A microbiological analysis was performed to determine the frequency of isolation of microorganisms of infectious and environmental type in milk from a group of clinically healthy cows in two different types of milking (manual and mechanical). To each sample of milk was made bacteriological culture to determine the presence of microorganisms. Of 289 milk samples evaluated, 193 (66.78%) were positive for isolation of any type of pathogen, of which 81 (28.02%) samples came from manual milking and 112 (38.75%) belonged to mechanical milking, finding a higher percentage of isolation of bacterial pathogens in milks coming from mechanical milking system (p = 0.0236). The most isolated pathogen was the Arcanobacterium haemolyticum, A microorganism that forms part of the saprophytic flora of the animal, with an individual presence in 20.14% and in coinfection with other pathogens in 0.7% of the samples. The most common microorganism of subclinical mastitis in cattle is Streptococcus agalactiae, which in the present study was isolated from 12.50% of milk samples. The odds ratio (OR) between the isolation of Streptococcus agalactiae and the Somatic Cell Score (SCS) was determined, which was statistically significant, indicating that when this pathogen is present the SCS increases and the animals are more susceptible to mastitis.

Resumo O objetivo deste estudo foi determinar a frequência de isoLamento de microorganismos de tipo infecciosos e ambientaL no Leite de um grupo de vacas cLinicamente saudáveL em dois diferentes tipos de ordenha. Para este fim utiLizaram-se amostras do Leite de 289 vacas LocaLizadas em rebanhos de quatro municipaLidades do Norte de Antioquia com dois tipos diferentes de ordenha (manuaL e mecânica), a cada amostra do Leite foi feito um cuLtivo bacterioLógico para determinar a presença de microorganismos. De 289 amostras do Leite avaLiadas, 193 (66,78%) foram positivas para o isoLamento de quaLquer tipo de agente patogênico (oito diferentes agentes patogênicos isoLados), das quais 81 (28,02%) amostras eram de ordenha manuaL e 112 (38, 75%) foram de ordenha mecânica, encontrando uma maior percentagem de isoLamento dos agentes patogênicos bacterianos nos Leites provenientes do sistema de ordenha mecânica (p = 0,0236). O agente patogênico mais isoLado foi Arcanobacterium haemolyticum (Corinebacterium sp), microrganismo que faz parte da flora saprófita do animaL, presentes individuaLmente em 20,14% das amostras anaLisadas, e em reLação com outros agentes patogênicos o 0,7% das amostras. O microrganismo que sempre se reporta como o maior produtor de mastite subcLínica em gado é o Streptococcus agalactiae, um agente patogênico infeccioso, que neste estudo foi isoLado no 12,50% das amostras do Leite. Foi determinado o Odds ratio (OR) entre o isoLamento de Streptococcus agalactiae e Pontuação das céLuLas somáticas (SCS), e foi estatisticamente significativo, indicando que quando o agente patogênico está presente, o SCS aumenta e os animais são mais susceptíveis à mastite.

The application value of SCS scoring system in emergency department / 中华急诊医学杂志

Objective To investigate the clinical value of the simple clinical score (SCS) in Emergency Department.Methods A total of 655 patients with critically illness admitted from July 1,2011through August 31,2011 were enrolled to evaluate the clinical application of SCS in emergency rescue room by analysis of the relationship between SCS and outcome of patients,and the correlation between each factor of SCS and the risk of death. Results The higher SCS,the higher hazard ratio for death.Differences in the hazard ratio for death among groups with different scores of SCS were statistical significance ( P ＜ 0.01 ).According to Chi-square test,there were significant differences in each factor ( except body temperature &unable standing or need Home Health Aide) among the groups of SCS ( P ＜ 0.05 ). Binary logistic regression analysis of each factor showed that age,diabetes,coma,pulse,systolic pressure and respiratory rate had significant correlation with patient mortality. After logistic regression analysis,age,diabetes,coma,pulse,systolic pressure and respiratory rate are significantly related to the mortality of patients.Conclusions The SCS scoring system is useful to make a precise evaluation of critically ill patients in the emergency department.When emergency rescue is carried out,particular stress should be focused on age,diabetes,coma,pulse,systolic pressure and respiratory rate.

Spent culture supernatant of Lactobacillus acidophilus influences contraction of isolated intestinal smooth muscle of rabbits / 第二军医大学学报

Objective: To observe the influence of spent culture supernatant (SCS) of Lactobacillus acidophilus strain LA14 on the contraction of isolated intestinal smooth muscle and to discuss the related mechanism. Methods: The ileum samples of rabbits were prepared and the contraction frequency and amplitude of intestinal smooth muscle were observed as the normal control. Then the SCS, bacterium suspension, and SCS with bacterium suspension were added by an accumulative dose to the culture media (0.3 ml per times, at an interval of 6 min), respectively. Four minutes after each administration, the contractive curves were recorded for 2 min. The influences of various groups of Lactobacillus acidophilus on the contraction of isolated intestinal smooth muscle were observed. The effect of SCS on M cholinoceptor was observed by adding in order pilocarpine, atropine or SCS, and pilocarpine. Results: After continuous administration of SCS or SCS with bacterium suspension (0.6-1.5 ml), the contraction frequency of the intestinal smooth muscle was significantly lowered compared with before administration (P0.05). Within the range of 0.3-1.5 ml, the SCS, bacterium suspension, and SCS with bacterium suspension resulted in no significant difference in reducing the contraction amplitude, except for SCS with bacterium suspension at 1.5 ml(P<0.05). SCS or atropine significantly inhibited pilocarpine-induced increase of contraction amplitude(P<0.05 or P<0.01). SCS also reduced the contraction frequency of the intestinal smooth muscle(P<0.01). Conclusion: SCS of Lactobacillus acidophilus may inhibit the peristalsis of the intestinal smooth muscle of rabbits by blocking M cholinoceptor.

Improved protocol for schwann cell isolation and proliferation from rat dorsal root ganglia in vitro