Objective To determine the effect of willed movement on the expression of Nogo-A and Rho-associated kinase (ROCK) in adult rats with cerebral ischemia.Methods Cerebral ischemia/reperfusion injury was established by middle cerebral artery occlusion (MCAO) for 2 h,followed by a 24 h reperfusion in 54 adult rats and the degree of their neurological deficit was evaluated using Longa scale.They were then divided randomly into 3 groups,namely the MCAO group,the environmental modification (EM) group,and the willed movement (WM) group.The rats of MCAO group were raised in a regular breeding box,where they could get food and water freely.Meanwhile,those of the other two groups were raised in a homemade box.For the WM group,the water bottle and food were located on the roof of the homemade box.In each group,six rats were killed on day 3,7 and 15 after reperfusion and their neurological deficits were evaluated respectively.Immunohistochemistry assay was employed to examine the expression of Nogo-A and ROCK in the brain tissue around the ischemic foci.Results The rats of the WM group showed lessened neurological deficits on day 15 compared with the model and EM group.Their expression of Nogo-A decreases from(28.92 ± 2.17)/hpf on day 7 to (24.38 ± 2.29)/hpf on day 15 and that of ROCK did from (40.03 ± 2.14)/hpf to (38.08 ± 2.07) / hpf,lower than those of the model and EM group.However,no significant differences were found in the expression of Nogo-A and ROCK between the model group and EM group at any time points.Conclusion Willed movement could promote the functional recovery of neurological deficits in rats with ischemia after reperfusion,which is probably in relation to restrained expression of Nogo-A and Rho-associated in the tissue around the brain ischemic foci.

Objective To investigate the relationship between low serum calcium concentration and hematoma volume in patients with intracerebral hemorrhage.Methods Between January 2012 and October 2014,870 consecutive patients with intracerebral hemorrhage admitted to the Department of Neurosurgery,West China Hospital,Sichuan University were enrolled prospectively.The patients completed laboratory serum calcium concentration and head CT examinations within 24 h after attack,and the baseline data and laboratory findings were collected.According to the normal reference value of laboratory serum calcium concentration,the patients were divided into a hypocalcemia calcium group (<2.1 mmol/L;n=193) and a normal calcium group (2.1-2.7 mmol/L;n=677).Spearman correlation analysis was used to analyze the correlation between the blood serum calcium concentration and the hematoma volume on admission.Results (1) The hypocalcemia group compared with normal calcium group,the proportion of male patients was high (73.6% [n=142] vs.66.0% [n=447]),the median score for Glasgow coma scale was lower (9 vs.11),and the median hematoma volume was larger (33.86 cm3 vs.21.69 cm3).The differences were statistically significant (all P<0.05).(2) Spearman correlation analysis showed that the lower serum calcium level on admission was weakly negatively correlated with the volume of hematoma in patients with intracerebral hemorrhage (r=-0.113,P<0.01).Conclusion The study suggested that the hypocalcemia on admission was mostly males in patients with intracerebral hemorrhage,the condition was serious,the volume of hematoma was larger,and the lower serum calcium concentration was negatively correlated with the hematoma volume.

OBJECTIVETo observe the changes of miRNA expression profiles in APPswe/PSδE9 transgenic mice and explore the possible roles of miRNA in the pathogenesis of Alzheimer's disease.

METHODSUsing miRNA chip technique, we examined the miRNA expression in the brain tissue of 6-month-old APPswe/PSδE9 transgenic mice, with age-matched wild-type mice as the control group.

RESULTSTwelve miRNAs showed differential expressions by more than two folds in APPswe/PSδE9 transgenic mice, namely miRNA-135a, miRNA-135a-2*, miRNA-298, miRNA-466b-3p, miR-669-3p, miR-142-5p, miR-144, miR-466f-3p, miR-466g, miR-200a, miR-200b and miR-96. Five miRNAs were significantly down-regulated in the transgenic mice, including miRNA-135a, miRNA-135a-2*, miRNA-298, miRNA-466b-3p, and miR-669-3p.

CONCLUSIONThe 5 down- regulated miRNA may play important roles in the pathogenesis of AD in APPswe/PSδE9 transgenic mice.

Alzheimer Disease ; genetics ; metabolism ; Animals ; Disease Models, Animal ; Gene Expression Profiling ; Mice ; Mice, Transgenic ; MicroRNAs ; genetics ; metabolism ; Oligonucleotide Array Sequence Analysis ; Transcriptome

Objective To establish a cell model mimicking Alzheimer's disease (AD) by knocking down SORL1 gene and compare the viability, apoptosis, and expressions of tumor necrosis factor-α( TNF-α) and interleukin-1β(IL-1β) in this model with a traditional Alzheimer's disease cell model. Methods A traditional cell model of AD was established by inducing N2a cells with Aβ25-35, and the optimal Aβ25-35 concentration was determined by assessing the cell viability changes. Another cell model of AD was established by transfecting N2a cells with SORL1-shRNA lentiviral vector, and SORL1 expression in the transfected cells were detected using Western blotting and qRT-PCR. With wild-type N2a cells without any treatment and cells transfected with a scramble shRNA as the control groups, the two cell models were examined for cell viability with MTT assay, cell apoptosis with flow cytometry, and TNF-αand IL-1βlevels in the culture supernatant with ELISA. Results The two cell models of AD showed obviously decreased viability and increased cell apoptosis compared with the untreated control cells or cells transfected with a scramble shRNA (P<0.05); no significant difference was found in the cell viability and apoptosis rate between the two AD cell models or between the two control groups (P>0.05). Significantly increased expressions of TNF-αand IL-1βwere observed in both of the two cell models compared with their respective control groups (P<0.05) without significant differences between the two cell models or between the two control groups (P>0.05). Conclusion A new AD cell model similar to Aβ25-35-induced AD model can be established by SORL1 knockdown in N2a cells.

To investigate the mechanism of Wenshen Xuanbi Decoction (WSXB) in treating osteoarthritis (OA) via network pharmacology, bioinformatics analysis, and experimental verification. The active components and prediction targets of WSXB were obtained from the TCMSP database and Swiss Target Prediction website, respectively. OA-related genes were retrieved from GeneCards and OMIM databases.Protein-protein interaction and functional enrichment analyses were performed, resulting in the construction of the Herb-Component-Target network. In addition, differential genes of OA were obtained from the GEO database to verify the potential mechanism of WSXB in OA treatment. Subsequently, potential active components were subjected to molecular verification with the hub targets. Finally, we selected the most crucial hub targets and pathways for experimental verification in vitro. The active components in the study included quercetin, linolenic acid, methyl linoleate, isobergapten, and beta-sitosterol. AKT1, tumor necrosis factor (TNF), interleukin (IL)-6, GAPDH, and CTNNB1 were identified as the most crucial hub targets. Molecular docking revealed that the active components and hub targets exhibited strong binding energy. Experimental verification demonstrated that the mRNA and protein expression levels of IL-6, IL-17, and TNF in the WSXB group were lower than those in the KOA group (p < 0.05). WSXB exhibits a chondroprotective effect on OA and delays disease progression. The mechanism is potentially related to the suppression of IL-17 and TNF signaling pathways and the down-regulation of IL-6.

Objective To establish a cell model mimicking Alzheimer's disease (AD) by knocking down SORL1 gene and compare the viability, apoptosis, and expressions of tumor necrosis factor-α( TNF-α) and interleukin-1β(IL-1β) in this model with a traditional Alzheimer's disease cell model. Methods A traditional cell model of AD was established by inducing N2a cells with Aβ25-35, and the optimal Aβ25-35 concentration was determined by assessing the cell viability changes. Another cell model of AD was established by transfecting N2a cells with SORL1-shRNA lentiviral vector, and SORL1 expression in the transfected cells were detected using Western blotting and qRT-PCR. With wild-type N2a cells without any treatment and cells transfected with a scramble shRNA as the control groups, the two cell models were examined for cell viability with MTT assay, cell apoptosis with flow cytometry, and TNF-αand IL-1βlevels in the culture supernatant with ELISA. Results The two cell models of AD showed obviously decreased viability and increased cell apoptosis compared with the untreated control cells or cells transfected with a scramble shRNA (P<0.05); no significant difference was found in the cell viability and apoptosis rate between the two AD cell models or between the two control groups (P>0.05). Significantly increased expressions of TNF-αand IL-1βwere observed in both of the two cell models compared with their respective control groups (P<0.05) without significant differences between the two cell models or between the two control groups (P>0.05). Conclusion A new AD cell model similar to Aβ25-35-induced AD model can be established by SORL1 knockdown in N2a cells.

A rapid analysis method based on ultraviolet-visual(UV-Vis) spectroscopy, near infrared(NIR) spectroscopy and multivariable data analysis was established for quality evaluation of Shengxuebao Mixture. The contents of eight active ingredients of Shengxuebao Mixture including albiflorin, paeoniflorin, 2, 3, 5, 4'-tetra-hydroxy-stilbene-2-O-β-D-glucopyranoside, specnuezhenide,ecliptasaponin D, emodin, calycosin-7-glucoside and astragaloside Ⅳ were simultaneously detected by using this method. HPLC-UV-MS was used as a reference method for determining the contents of these ingredients. Partial least squares(PLS) analysis was implemented as a linear method for multivariate models calibrated between UV spectrum/NIR spectrum and contents of 8 ingredients. Finally, the performance of the model was evaluated by 24 batches of test samples. The results showed that both UV-Vis and NIR models gave a good calibration ability with an R~2 value above 0.9, and the prediction ability was also satisfactory, with an R~2 value higher than 0.83 for UV-Vis model and higher than 0.79 for NIR model. The overall results demonstrate that the established method is accurate, robust and fast, therefore, it can be used for rapid quality evaluation of Shengxuebao Mixture.
Calibration ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Least-Squares Analysis ; Mass Spectrometry ; Spectroscopy, Near-Infrared

Objective:To study the safety and efficacy of endoscopic papillary balloon dilation for choledocholithiasis.Methods:A total of 60 patients with choledocholithiasis in Suqian People′s Hospital of Nanjing Drum Tower Hospital Group were included from January 2017 to December 2018 according to the inclusion and exclusion criteria. According to the random number table, the patients were divided into two groups: simple endoscopic papillary balloon dilation group (EPBD group, n=30) and endoscopic papillary sphincterotomy combined with balloon dilation group (ESBD group, n=30). Lithotripsy time, X-ray exposure time, one-time lithotripsy rate, lithotripsy rate, incidence of postoperative acute pancreatitis, intraoperative and postoperative bleeding rates were compared.Results:The time of stone extraction (8.5±2.4 min) in EPBD group was comparable with that of group ESBD (7.8±2.1 min) ( P=0.14). The time of X-ray exposure was 21.8±5.2 min in EPBD group and 19.7±6.3 min in ESBD group ( P=0.11). Stones were extracted at one time in all 60 patients, and no lithotripsy was conducted. The incidences of acute pancreatitis after endoscopic retrograde cholangiopancreatography(ERCP) in the two groups were both 6.67% (2/30). The intraoperative bleeding rates were 3.33% (1/30) and 10.00% (3/30) in EPBD group and ESBD group ( P=0.042), respectively. The rate of postoperative bleeding was 3.33% (1/30) in ESBD group ( P=0.035). No other recent complications occurred in the two groups. Conclusion:Endoscopic papillary balloon dilation alone is safe and effective in the treatment of choledocholithiasis.

To study the preventive effects of double guidewire technique combined with pancreatic duct stenting in preventing post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis (PEP). Patients receiving ERCP were divided into the treatment group and the control group by random number table. In the treatment group, double guidewire technique combined with pancreatic duct stenting was applied. In the control group, selective biliary intubation was applied in the conventional way. The intubation time, PEP, hyperamylasemia and bleeding incidence were analyzed between the two groups. A total of 80 patients were enrolled in this study from January 2016 to December 2018. There were 40 cases in the treatment group and 39 cases in the control group. In the treatment group, the mean intubation time was 384±102 seconds. No PEP or bleeding during and after the operation occurred, but hyperamylasemia occurred in 2 cases. In the control group, the mean intubation time was 427±115 seconds. Hyperamylasemia occurred in 6 cases, PEP occurred in 3 cases, and 1 case of intraoperative bleeding happened in the control group. The incidence of PEP [0 VS 7.7%(3/39)]and hyperamylasemia [5.0% (2/40)VS 15.4%(6/39)] were lower in the treatment group (both P<0.05). Double guidewire technique combined with pancreatic duct stenting can successfully perform selective bile duct intubation and effectively prevent PEP.

In this study, the HPLC-UV-MS method for the simultaneous determination of eight active ingredients of Shengxuebao Mixture were developed based on the concept of quality by design（QbD）with a stepwise optimization approach. After the analytical target profile（ATP）had been defined, albiflorin, paeoniflorin, 2, 3, 5, 4'-tetra-hydroxy-stilbene-2-O-β-D-glucopyranoside, specnuezhenide, ecliptasaponin D, emodin, calycosin-7-glucoside, and astragaloside Ⅳ were identified as the indicator components. The resolution and the signal-to-noise ratio of indicator components were then selected as critical method attributes (CMA) for the first step optimization. According to the results collected from fractional factorial design, critical method parameters (CMP) were determined with a multiple linear regression method, which included the amount of acid addition in the mobile phase, temperature, gradient, and wavelength. After that, the amount of acid addition and the wavelength were optimized to improve the resolution and the signal-to-noise ratio of the indicator components. The peak symmetry factors of specnuezhenide and emodin were then set as CMA for the second step optimization. The Box-Behnken designed experiments were conducted. The temperature and gradient were optimized after modelling. The design space were calculated and verified. The optimized analytical method was validated, and the results showed a good precision, accuracy and stability, which means that it can be used for the quantification of the indicator components in Shengxuebao Mixture.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Mass Spectrometry ; Phytochemicals ; analysis ; Reproducibility of Results