Improvement effect of crocin on cognitive impairment of Alzheimer's disease rats through DKK3 regulation of GSK-3 β/β-Catenin pathway / 中国临床药理学与治疗学

AIM: To explore the improvement effect and mechanism of crocin on cognitive impairmrnt of Alzheimer's disease (AD) rats. METHODS: The hippocampus of SD rats were injected with Aβ 25-35 to establish AD model, then rats were randomly divided into AD group, AD + low, medium, high dose of crocin groups (10, 20, 40 mg/kg) and AD + donepezil group (1 mg/kg), intraperitoneal injection treatment for 4 weeks, set sham group. Dark avoidance test and water maze test were used to evaluate the learning and memory abilities of rats, ELISA was used to detect serum Aβ content, HE staining and Tunel staining were used to determine pathological changes and neuronal apoptosis of hippocampus of rats, immunohistochemistry was used to detect the expression of Brdu, Dcx and NeuN in hippocampus of rats, and Western blot was used to detect the protein expression of Aβ, DKK3, β-catenin, p-GSK-3β/GSK-3β, Caspase-3, Bax, Bcl-2 in hippocampus of rats. RESULTS: Compared to sham group, the learning and memory abilities of AD group rats were decreased, serum Aβ content increased, the pathological change in hippocampus was serious, neuronal apoptosis was increased, the expression of Brdu, Dcx, NeuN were decreased, the protein expression of Aβ, DKK3, p-GSK-3β/GSK-3β, Caspase-3, Bax were increased, protein expression of β-catenin, Bcl-2 were decreased (P<0.01). Compared to AD group, after the treatment of doses of crocin and donepezil, the learning and memory abilities of AD rats were improved, serum Aβ content were increased, and the pathological change in hippocampus were alleviated, neuronal apoptosis were reduced, the expression of Brdu, Dcx, NeuN were decreased, the protein expression of Aβ, DKK3, p-GSK-3β/ GSK-3β, Caspase-3, Bax were decreased, the protein expression of β-catenin, Bcl-2 were increased, notely, dose-dependent effect of crocin was significant. CONCLUSION: Crocin reduced neuronal apoptosis and mediated DKK3 to regulate GSK-3β/ β-catenin pathway to improve the cognitive impairment of AD rats.

Control of lupus activity during pregnancy via the engagement of IgG sialylation: novel crosstalk between IgG sialylation and pDC functions / 医学前沿

Immunoglobulin (IgG) glycosylation affects the effector functions of IgG in a myriad of biological processes and has been closely associated with numerous autoimmune diseases, including systemic lupus erythematosus (SLE), thus underlining the pathogenic role of glycosylation aberration in autoimmunity. This study aims to explore the relationship between IgG sialylation patterns and lupus pregnancy. Relative to that in serum samples from the control cohort, IgG sialylation level was aberrantly downregulated in serum samples from the SLE cohort at four stages (from preconception to the third trimester of pregnancy) and was significantly associated with lupus activity and fetal loss during lupus pregnancy. The type I interferon signature of pregnant patients with SLE was negatively correlated with the level of IgG sialylation. The lack of sialylation dampened the ability of IgG to suppress the functions of plasmacytoid dendritic cells (pDCs). RNA-seq analysis further revealed that the expression of genes associated with the spleen tyrosine kinase (SYK) signaling pathway significantly differed between IgG- and deSia-IgG-treated pDCs. This finding was confirmed by the attenuation of the ability to phosphorylate SYK and BLNK in deSia-IgG. Finally, the coculture of pDCs isolated from pregnant patients with SLE with IgG/deSia-IgG demonstrated the sialylation-dependent anti-inflammatory function of IgG. Our findings suggested that IgG influences lupus activity through regulating pDCs function via the modulation of the SYK pathway in a sialic acid-dependent manner.

Related factors of endogenous erythropoietin and its association with 10-year risks of cardiovascular disease in a community-based Chinese study / 北京大学学报(医学版)

OBJECTIVE@#To investigate the associated factors of endogenous erythropoietin (EPO) and its association with 10-year risks of atherosclerotic cardiovascular disease in a Chinese community-based general population.@*METHODS@#The participants of this study were from an atherosclerosis cohort survey which was established by the Department of Cardiology, Peking University First Hospital in 2011. The cohort survey was performed in the Gucheng and Pingguoyuan communities of Shijingshan district in Beijing, China. The inclusion criteria of this study were: (1) endogenous EPO was measured; (2) health questionnaire data and other clinical data were complete; (3) participatants who had cardiovascular or cerebrovascular diseases (defined as self-reported coronary heart disease, stroke or transient ischemic attack) or anemia or estimated glomerular filtration rate (eGFR) < 60 mL/(min·1.73 m2) at baseline were excluded. Multivariate linear regression model was used to examine the associated factors of endogenous EPO. The participants were grouped into low (< 5%), moderate (5%-10%) and high risk (≥10%) groups, based on predicted 10-year cardiovascular disease risk using the prediction for atherosclerotic cardiovascular disease risk in China (China-PAR) equations.@*RESULTS@#A total of 4 013 participants were included. Mean age of them was (55.9±8.2) years, 62.2% (n=2 496) of them were female, and 46.3% (n=1 859), 70.9% (n=2 845), 21.9% (n=879) had hypertension, dyslipidemia and diabetes, individually. The average body mass index was (26.1±3.3) kg/m2. The median of EPO level was 12.8 (9.3-17.4) IU/L and 25.1% (n=998) were at high 10-years risk of cardiovascular disease. Hemoglobin (β=-0.05, 95%CI: -0.07 to -0.04) and eGFR ≥90 mL/(min·1.73 m2) (β=-0.05, 95%CI: -0.07 to -0.04) were associated with lower in transformed EPO levels while hypertension (β=0.08, 95%CI: 0.05 to 0.12) and obesity (β=0.14, 95%CI: 0.09 to 0.18) were associated with higher in transformed EPO levels in multivariate linear regression analyses. Ten-year cardiovascular disease risks were positively associated with in transformed EPO levels (β=0.07, 95%CI: 0.05 to 0.09). The participants at moderate and high cardiovascular disease risks had significant higher EPO levels than the low risk group (all P < 0.05).@*CONCLUSION@#In community-based Beijing populations, endogenous EPO was associated with hemoglobin, renal function, obesity and hypertension. Individuals at high 10-years cardiovascular disease risks have higher endogenous EPO levels. Endogenous EPO may be a potential risk marker of cardiovascular disease.

Development of evaluation index system for community screening and referral procedure of patients with chronic heart failure / 中华全科医师杂志

Objective:To develop an evaluation index system for community screening and referral procedure of patients with chronic heart failure.Methods:Experts in fields of medical education, health administration and various clinical specialties were selected from Shanghai through the purpose sampling method. Two rounds of expert consultation with Delphi method were conducted during October 2020 to February 2021, the contents of consultation included the importance and availability of the index system. According to the consultation results, the index system for community screening and referral procedure of patients with chronic heart failure was developed.Results:A total of 16 experts participated in the consultation, among whom 15 held senior or associate senior titles and 14 had worked for 20 years. The recovery rates of valid questionnaires of two rounds expert of consultation were 16/16. The familiarity and judgment coefficient were 0.82 and 0.90 in the first round of expert consultation, and 0.86 and 0.90 in the second round. The expert consultation coordination coefficient of the importance in the community screening and referral procedure were 0.652 and 0.462 for the first consultation; 0.741 and 0.525 for the second consultation. In the final version of the evaluation index system there were 7 first-level indicators, 15 secondary indicators and 3 conditions for screening; and 2 first-level indicators, 5 secondary indicators and 1 condition for referral.Conclusion:In this study the positivity coefficient and authority degree of experts are high; the opinions are relatively concentrated. The consultation results have a high rationality and feasibility, which would be applicable for assessment of community screening and referral procedure of patients with chronic heart failure.

Role of protein O-GlcNAc modification in oxidative stress injury in nerve cells of mice subjected to oxygen-glucose deprivation and restoration / 中华麻醉学杂志

Objective:To evaluate the role of protein O-linked beta-N-acetylglucosaminylation (O-GlcNAcylation) modification in oxidative stress injury in nerve cells of mice subjected to oxygen-glucose deprivation and restoration (OGD/R).Methods:The standard mouse hippocampal neuron cell line was inoculated on a culture plate or dish at a density of 5×10 4 cells/ml and divided into 4 groups ( n=20 each) using a random number table method: normal group (N group), O-(connection)N-acetylglucosamine hydrolase (OGA) inhibitor Thiamet G group (T group), OGD/R group (D/R group) and Thiamet G+ OGD/R complex sugar group (T-D/R group). The cells were exposed to a mixed gas of 94% N 2-5% CO 2-1% O 2 for 6 h in a low-glucose medium, then medium was replaced with a common medium for restoring oxygen and glucose, and the cells were cultured for 12 h. Thiamet G at a final concentration of 1 mmol/L was added to the culture medium at 4 h before OGD/R in T-D/R group, and the medium was replaced with a medium containing Thiamet G at a final concentration of 1 mmol/L at 4 h before extraction of cellular proteins.After oxygen and glucose restoration was completed, the accumulation of cellular ROS was measured using DCFH-DA staining, mitochondrial membrane potential was measured using Jc-1 staining, O-GlcNAc modification was determined by immunofluorescence, and the expression of nuclear factor E2-related factor 2 (Nrf2), c-Jun N-terminal kinase (JNK), phosphorylated JNK (p-JNK), and p53 tumor suppressor gene (p53) was detected using Western blot. Results:Compared with group N, the expression of O-GlcNAc in nerve cells was significantly up-regulated in group T, and the accumulation of ROS in nerve cells was significantly increased, JC-1 monomer was increased, JC-1 polymer was decreased, Nrf2 expression was down-regulated, and the expression of p-JNK and p53 was up-regulated in group D/R, and the expression of O-GlcNAc in nerve cells was up-regulated, the accumulation of ROS was increased, the polymerization of JC-1 monomer and JC-1 was increased, Nrf2 expression was down-regulated, and the expression of p-JNK and p53 was up-regulated in group T-D/R ( P<0.05). Compared with group D/R, the expression of O-GlcNAc in nerve cells was significantly up-regulated, the accumulation of ROS was decreased, JC-1 monomer was decreased, JC-1 polymer was increased, the expression of Nrf2 was up-regulated, and the expression of p-JNK and p53 was down-regulated in group T-D/R ( P<0.05). Conclusion:When mouse nerve cells are subjected to OGD/R, the protein O-GlcNAc modification as an endogenous protective mechanism is enhanced, which can reduce oxidative stress injury, and the mechanism may be related to regulating the Nrf2-mediated JNK pathway.

Advance of neurodegeneration in diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

@#Diabetic retinopathy(DR), one of the most common complications of diabetes, has been widely reported as microangiopathy. However, retinal neurodegeneration was reported to occur early in DR and played a significant role in DR progression. Retinal neurodegeneration in DR was characterized as neuronal apoptosis and reactive gliosis. The mechanisms for its pathogenesis include hyperglycemia, oxidative stress, glutamate excitotoxicity, and inflammation <i>etc</i>. Furthermore, retinal neurodegeneration has a close relationship with the microangiopathy in the pathogenesis of DR.

Medical treatment of ocular fundus disease and challenges / 中华眼底病杂志

Fundus disease is a kind of common ophthalmic disease with high blindness rate and great harm. Although great breakthroughs have been made in medical treatment, laser photocoagulation, radiotherapy and gene therapy of fundus disease, with the further understanding of the essence of fundus disease, there are higher requirements for the treatment of fundus disease. To strengthen scientific research on the etiology, pathological mechanism and clinical treatment of fundus diseases, to control the quality of research, to develop reasonable treatment plans and explore new treatment methods under the guidance of evidence-based medicine theory, to further improve the level of medical treatment and benefit patients with fundus diseases will still be a formidable challenge in the future.

Relationship between PERK pathway-mediated endoplasmic reticulum stress and reduction of cerebral ischemia-reperfusion injury by dexmedetomidine in mice / 中华麻醉学杂志

Objective:To evaluate the relationship between protein kinase RNA-like endoplasmic reticulum kinase (PERK) pathway-mediated endoplasmic reticulum stress and the reduction of cerebral ischemia-reperfusion (I/R) injury by dexmedetomidine in mice by the in vivo experiment and the cell experiment. Methods:In the in vivo experiment, 20 healthy clean-grade male mice, aged 6-8 weeks, weighing 20-30 g, were divided into 4 groups ( n=5 each) using a random number table method: sham operation group (group S), sham operation+ dexmedetomidine group (group SD), cerebral I/R group (group IR) and cerebral I/R+ dexmedetomidine group (group IRD). Cerebral I/R was established by two-vessel occlusion plus hypotension.Dexmedetomidine 25 μg/kg was intraperitoneally injected at 10 min of ischemia in group IRD and at the corresponding time point in group SD.Neurological function was assessed using modified neurological severity score at 1 h of reperfusion.The animals were then sacrificed and brain tissues were taken for determination of the expression of endoplasmic reticulum stress-related proteins such as immunoglobulin heavy chain-binding protein (BIP), eukaryotic translation initiation factor 2α (EIF-2α), phosphorylated EIF-2α (p-EIF-2α), PERK and phosphorylated PERK (p-PERK) (by Wester blot). In the cell experiment, a mouse hippocampal neuronal cell line was selected and divided into 4 groups ( n=12 each) using a random number table method: control group (group C), oxygen-glucose deprivation/restoration (OGD/R) group (group OGD/R), OGD/R+ dexmedetomidine group (group OGD/R+ D) and OGD/R+ ISRIB (PERK pathway inhibitor) group (group OGD/R+ ISRIB). Cells were exposed to 94%N 2-5%CO 2-1%O 2 and incubated in a low-glucose DMEM medium for 6 h followed by restoration to establish OGD/R model.At 30 min before OGD, dexmedetomidine (final concentration 5 mmol/L) was added in group OGD/R+ D, and ISRIB (final concentration 10 mmol/L) was added in group OGD/R+ ISRIB.After 12-h restoration was completed, the cell survival rate was detected by CCK-8 assay.At 24 of restoration, the expression of endoplasmic reticulum stress-related proteins was determined by Wester blot. Results:In the in vivo experiment, compared with group S, neurobehavioral score was significantly increased and the expression of BIP, p-EIF-2α and p-PERK in brain tissues was up-regulated in group IR ( P<0.05). Compared with group IR, neurobehavioral score was significantly decreased and the expression of BIP, p-EIF-2α and p-PERK in brain tissues was down-regulated in group IRD ( P<0.05). In the cell experiment, compared with group C, the expression of BIP, p-EIF-2α, PERK and p-PERK was significantly up-regulated, and the cell survival rate was decreased in group OGD/R ( P<0.05). Compared with group OGD/R, the expression of BIP, p-EIF-2α, PERK and p-PERK was significantly down-regulated, and the cell survival rate was increased in OGD/R+ D, OGD/R+ ISRIB groups ( P<0.05). Compared with group OGD/R+ ISRIB, the expression of PERK was significantly down-regulated ( P<0.05) and no significant change was found in the other parameters in group OGD/R+ D ( P>0.05). Conclusion:The mechanism by which dexmedetomidine reduces cerebral I/R injury may be related to inhibiting PERK pathway-mediated endoplasmic reticulum stress in mice.

Research progress on bilingualism and Alzheimer's disease / 中华老年医学杂志

Alzheimer's disease(AD)is the most common type of dementia.It is of great significance to systematically explore the interaction between bilingualism and AD for patients' cognitive rehabilitation and reduction of the burden on patients' families and society.Based on our literature review, bilingualism may be a beneficial factor for cognitive reserve helping to delay the onset of AD and thus exert positive effects mitigating the decline in patients' cognitive ability such as memory, executive control and visuospatial function.In turn, AD can impair patients' bilingual ability to varying degrees.In future research, more efforts should be directed toward elucidating the influence of bilingual characteristics, the age of second language acquisition, second language proficiency and other factors on AD as well as the influence of AD on patients' attention, emotional cognition and social cognition.

Standard Operating Procedures for Chinese Medicine Data Monitoring Committees of Clinical Studies / 中国结合医学杂志

Although there is guidance from different regulatory agencies, there are opportunities to bring greater consistency and stronger applicability to address the practical issues of establishing and operating a data monitoring committee (DMC) for clinical studies of Chinese medicine. We names it as a Chinese Medicine Data Monitoring Committee (CMDMC). A panel composed of clinical and statistical experts shared their experience and thoughts on the important aspects of CMDMCs. Subsequently, a community standard on CMDMCs (T/CACM 1323-2019) was issued by the China Association of Chinese Medicine on September 12, 2019. This paper summarizes the key content of this standard to help the sponsors of clinical studies establish and operate CMDMCs, which will further develop the scientific integrity and quality of clinical studies.

Induction chemotherapy for unresectable Stage III non-small-cell lung cancer may improve survival of induction chemotherapy responders as predicted by elevated levels of carcinoembryonic antigen and cytokeratin fragment 19 and classification as stage N3 cancer

Aims: The aim of this study is to investigate patients with unresectable Stage III non-small-cell lung cancer (NSCLC) receiving radiotherapy with induction and concurrent pemetrexed or docetaxel plus cisplatin (PP/DP) chemotherapy and to identify the subgroup most likely to benefit from induction chemotherapy (IC). Subjects and Methods: Patients with unresectable measurable Stage III NSCLC received two cycles of PP/DP IC followed by concurrent chemoradiotherapy at a dose of 60–66 Gy. Statistical Analysis Used: Cox regression analysis was performed to evaluate the prognostic factors for survival; logistic regression analysis was used to evaluate the predictors for response to IC, and the receiver operating characteristic curves were used to evaluate the independent factors predicting response. Results: Eighty patients were included; the median survival time (MST) was 22.1 months. Partial response (PR) to IC was an independent prognostic factor for overall survival. For patients in the PR and stable disease groups, the MST was 36.7 and 19.5 months, respectively. The independent predictors of PR to IC included classification as stage N3 cancer, baseline carcinoembryonic antigen (CEA) levels >10 ng/ml, and cytokeratin fragment 19 (CYFRA21-1) levels >6 ng/ml. With each additional independent predictor, the likelihood of having have PR to IC increased. Conclusions: Radiotherapy with induction and concurrent PP/DP chemotherapy is feasible for patients with unresectable Stage III NSCLC. IC may improve the survival of IC responders, as predicted by elevated CEA and CYFRA21-1 levels and classification as stage N3 cancer. Additional randomized trials on IC may consider these predictors to tailor individualized treatments

Clinical features of 9 cases of Brucella endocarditis / 中华心血管病杂志

Objective: To explore the clinical characteristics of patients with Brucella endocarditis. Methods: The clinical data of 9 patients with Brucella endocarditis admitted to Beijing Ditan Hospital from October 2008 to August 2018 were retrospectively analyzed. Through the electronic medical record system of the hospital. Through assessing the electronic medical record system of the hospital, demographic data, main symptoms, vital signs, blood culture, Rose Bengal Plate Agglutination Test, echocardiography, electrocardiogram, chest imaging and other clinical data of included patients were inquired and recorded. Patients were followed up by telephone for medication, operation and outcome. Results: The 9 patients were all Han nationality, aged from 25 to 66 years, 7 out of 9 patients were male, and they came from Hebei, Shandong, Shanxi, Inner Mongolia and Beijing. Of the 9 patients, 5 were farmers, 2 were self-employed, 1 was a technician, and 1 was unemployed. Of the 9 cases, 8 had a history of close contact with cattle and sheep, and 5 had a history of eating beef and mutton. Rose-Bengal Plate Agglutination Test and blood culture were positive in all 9 patients. Aortic valve was involved in 7 out of 9 patients, mitral and tricuspid valve was involved in 1 patient, respectively, and aortic dissection occurred in 1 patient. Condition of 1 patient rapidly deteriorated after admission and finally died during hospitalization despite antibiotic therapy, the remaining patients received long-term antibiotic treatment. A total of 7 patients who underwent valve replacement were followed up. One patient died of cerebral hemorrhage 6 months after operation, and the remaining 6 patients recovered well after valve replacement. Heart failure occurred in all 9 patients, and pericardial effusion occurred in 8 patients. Electrocardiogram showed low voltage of the QRS complex in the limb in 3 cases and poor R-wave progression in V(1)-V(3) lead in 2 cases, and sinus tachycardia in 2 cases. One patient developed non-specific ST-T abnormalities. All patients had fever, 7 patients complained of weakness, and 6 patients complained of palpitations. Among the 9 patients, 7 cases had anemia, 7 patients had pneumonia, 6 had bilateral pleural effusion, 4 had thrombocytopenia. Creatinine was above normal in 4 patients, urine protein was positive in 3 patients, Delta Bilirubin was higher in 3 cases. Conclusions: Patients with Brucella endocarditis often suffer from heart failure and have severe complications. Adequate antibiotic therapy in combination with valve replacement is effective for the treatment of patients with Brucella endocarditis.

Gene editing: an instrument for practical application of gene biology to plant breeding / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Plant breeding is well recognized as one of the most important means to meet food security challenges caused by the ever-increasing world population. During the past three decades, plant breeding has been empowered by both new knowledge on trait development and regulation (e.g., functional genomics) and new technologies (e.g., biotechnologies and phenomics). Gene editing, particularly by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) and its variants, has become a powerful technology in plant research and may become a game-changer in plant breeding. Traits are conferred by coding and non-coding genes. From this perspective, we propose different editing strategies for these two types of genes. The activity of an encoded enzyme and its quantity are regulated at transcriptional and post-transcriptional, as well as translational and post-translational, levels. Different strategies are proposed to intervene to generate gene functional variations and consequently phenotype changes. For non-coding genes, trait modification could be achieved by regulating transcription of their own or target genes via gene editing. Also included is a scheme of protoplast editing to make gene editing more applicable in plant breeding. In summary, this review provides breeders with a host of options to translate gene biology into practical breeding strategies, i.e., to use gene editing as a mechanism to commercialize gene biology in plant breeding.

Investigation and analysis of somatic symptom disorder and anxiety and depression in patients with myocardial bridge / 中华行为医学与脑科学杂志

Objective:To investigate and analyze the somatic symptom disorder, anxiety and depression in patients with myocardial bridge.Methods:A total of 276 patients with myocardial bridge diagnosed by coronary angiography (CAG) were enrolled in the department of cardiology of Renji hospital in Shanghai from June to December in 2016. There were 151 cases of simple myocardial bridge (no coronary stenosis or coronary artery stenosis <30%) and 125 cases of complex myocardial bridge (combined with coronary stenosis >30%). A total of 1067 patients with myocardial bridge without coronary angiography were collected at the same time. Self-rating somatic symptom scale (SSS), generalized anxiety disorder (GAD-7) and patient health questionnaire (PHQ -9) were given to these patients during hospitalization. At the same time, somatic symptoms disorder and anxiety and depression in the myocardial bridge group and non-myocardial bridge group were compared.Results:The prevalence of somatic symptom disorder in patients with myocardial bridge was higher than that in non-myocardial bridge patients (35.86% vs 28.30%, P<0.05). There was significant correlation between somatic symptom disorder and depression and anxiety, with correlation coefficients of 0.629 and 0.565, respectively. The prevalence of depression and anxiety in myocardial bridge patients was higher than that in non-myocardial bridge patients (depression: 23.91% vs 22.11%. P=0.467; anxiety: 17.02% vs 14.15%, P=0.22), but there was no statistical difference. For male patients or female patients, the prevalence of somatic symptom disorder, depression and anxiety in the simple myocardial bridge patients were higher than those in the complex myocardial bridge patients, but there was no statistical difference. The most common non-specific somatic symptoms disorder in patients with myocardial bridge were fatigue (64.5%), followed by sleep disorders (63.8%) and decreased attention (63.0%). Conclusion:The somatic symptom disorder in patients with myocardial bridge is significantly higher than that in non-myocardial bridge group. Especially for patients with myocardial bridge with non-specific somatic symptoms, early identification of somatic symptoms disorder of myocardial bridge patients will be beneficial to proper clinical invitation.

Comparison of different methods for drug delivery via the lumbar spinal subarachnoid space in rats / 南方医科大学学报

OBJECTIVE@#To compare 3 commonly used methods for drug delivery via the lumbar spinal subarachnoid space in rats.@*METHODS@#We compared the effects of 3 methods for drug delivery via the lumbar spinal subarachnoid space in Sprague Dawley rats, namely acute needle puncture, chronic catheterization via laminectomy, and non-laminectomized catheterization. Body weight changes of the rats were measured, and their general and neurological conditions were assessed after the surgeries. The motor function of the rats was examined using rota rod test both before and after the surgeries. Nociceptive tests were performed to assess nociception of the rats. HE staining was used to examine local inflammation caused by the surgeries in the lumbar spinal cord tissue, and lidocaine paralysis detection and toluidine blue dye assay were used to confirm the precision of drug delivery using the 3 methods.@*RESULTS@#Both needle puncture and catheterization via laminectomy resulted in a relatively low success rate of surgery and caused neurological abnormalities, severe motor dysfunction, hyperalgesia, allodynia and local inflammation. Catheterization without laminectomy had the highest success rate of surgery, and induced only mild agitation, slight cerebral spinal fluid leakage, mild sensory and motor abnormalities, and minimum pathology in the lumbar spinal cord. Catheterization without laminectomy produced less detectable effects on the behaviors in the rats and was well tolerated compared to the other two methods with also higher precision of drug delivery.@*CONCLUSIONS@#Catheterization without laminectomy is a safe, accurate and effective approach to lumbar drug delivery in rats.

Optimization of the Extraction Technology of Verbascoside from Cistanche tubulosa by Box-Behnken Response Surface Me- thodology / 中国药房

OBJECTIVE： To optimize the extraction technology of verbascoside from Cistanche tubulosa， and to provide reference for further development and comprehensive utilization of C. tubulosa. METHODS： The content of verbascoside in C. tubulosa was determined by HPLC. The determination was performed on  Inertsil-ODS-3V  column with  mobile phase consisted of methanol-0.2％ formic acid aqueous solution （40 ∶ 60， V/V） at  the flow rate  of  1 mL/min. The column temperature was 30 ℃， the detection wavelength was 330 nm， and the sample size  was 10 μL. Using extraction rate of verbascoside as index， soaking time， ethanol concentration， liquid-solid ratio， extraction time and extraction times were investigated by single factor tests. According to the results of above tests， ethanol concentration， liquid-solid ratio and extraction time were optimized by Box-Behnken response surface methodology. The verification tests were carried out on the optimized extraction technology. RESULTS： The linear range of verbascoside was 18.65-932.4 μg/mL. The optimal extraction technology included that ethanol concentration 63％， liquid-solid ratio 8 ∶ 1 （mL/g）， soaking for 2 h， extraction time 1.5 h， extracting for 2 times. The extraction rates of verbascoside in the three parallel verification tests were 78.21％， 76.95％， 79.34％， respectively. The relative errors of those to predicted value 76.76％ were 1.89％， 0.25％， 3.36％. CONCLUSIONS： The optimized extraction technology of verbascoside from C. tubulosa is stable and feasible， and is suitable for the extraction of verbascoside.

Total flavone of abelmoschl manihot L. medic exhibits protective effect against hind-limb ischemia in rat model

Total flavones of Abelmoschus manihot L. Medic [TFA] is the major active component isolated from Chinese herb Abelmoschus manihot L. Medic. TFA has shown neuroprotective effect against cerebral ischemia injury in rats and rabbits. However, the effects of TFA on hind-limb ischemia and the underlying mechanisms remain unclear. Therefore, in the present study, we used a rat hind-limb ischemia model to investigate protective effect of TFA against limb ischemia injury. The rat model of hind-limb ischemia was established. Treatment groups received TFA at two different doses [160 and 40mg/kg] daily for 10 days. Sham operated control group and model group received saline. At the end the rats were sacrificed, hindlimb tissues were stained with Haematoxylin-Eosin and Masson's trichrome. RNA and protein were extracted from tissues for PCR and Western blot analysis. The results showed that TFA reduced lower limb ischemic injury, recovered tissue volume and diminished fibrosis and muscle degeneration. Mechanistically, we showed that TFA increased the expression of anti-apoptotic factor such as Bcl-2 and survivin, decreased the expression of pro-apoptotic factor such as Caspase 3, Bax and Bak and inhibited the activation of caspase 3 and 9. In summary, this study proves new evidence that TFA protects hind-limb against ischemia injury by inhibiting apoptosis and could be a promising therapeutic agent for acute lower extremity ischemia

Role of TIPE2 in macrophage pyroptosis in mice / 中华麻醉学杂志

Objective To evaluate the role of tumor necrosis factor alpha-inducible protein 8 like-2 ( TIPE2) in macrophage pyroptosis in mice. Methods Mouse macrophages J774A. 1 were seeded in 6-cm culture dishes (5 ml∕dish) at the density of 2×105 cells∕ml and divided into 4 groups (n=18 each) using a random number table method: blank vector control group ( C group) , blank vector plus lipopolysaccharide ( LPS)∕ATP group ( C+LPS∕ATP group) , TIPE2 overexpression group ( T group) and TIPE2 overexpres-sion plus LPS∕ATP group ( T+LPS∕ATP group) . Cells were infected with lentivirus without TIPE2 in C and C+LPS∕ATP groups. TIPE2 overexpression stable cell line was constructed in T group and T+LPS∕ATP group. LPS 1. 0 μg∕ml was added and cells were incubated for 5 h, and then ATP 5. 0 mmol∕L was added and cells were incubated for 1 h in C+LPS∕ATP group and T+LPS∕ATP group. Cells were collected for de-tection of the expression of TIPE2, NOD-like receptor protein 3 ( NLRP3) , interleukin-1beta ( IL-1β) and interleukin-8 ( IL-18) by Western blot. Flow cytometry was used to detect the pyroptotic cells, and the per-centage of pyroptotic cells was calculated. The concentrations of tumor necrosis factor-alpha ( TNF-α) , IL-6, IL-1β and IL-18 in cell culture media were determined by enzyme-linked immunosorbent assay. Results Compared with group C, the expression of TIPE2 was significantly down-regulated, the expression of NL-RP3, IL-1β and IL-18 was up-regulated, and the percentage of pyroptotic cells and concentrations of TNF-α, IL-6, IL-1β and IL-18 in cell culture media were increased in group C+LPS∕ATP (P<0. 05). Com-pared with group T, the expression of TIPE2 was significantly down-regulated, the expression of NLRP3, IL-1βand IL-18 was up-regulated, and the percentage of pyroptotic cells and concentrations of TNF-α, IL-6, IL-1β and IL-18 in cell culture media were increased in group T+LPS∕ATP ( P<0. 05) . Compared with group C+LPS∕ATP, the expression of TIPE2 was significantly up-regulated, the expression of NLRP3, IL-1β and IL-18 was down-regulated, and the percentage of pyroptotic cells and concentrations of TNF-α, IL-6, IL-1β and IL-18 in cell culture media were decreased in group T+LPS∕ATP ( P<0. 05) . Conclusion TIPE2 can inhibit macrophage pyroptosis, and the mechanism may be related to inhibiting activation of NL-RP3 inflammasome in mice.

Impact of Remote Ischemic Conditioning on Circulating miR-208b Level and Cardiac Function in Patients With ST Segment Elevation Myocardial Infarction Undergoing Primary Angioplasty / 中国循环杂志

Objectives: To investigate whether remote ischemic conditioning (RIC) applied to patients with ST-segment elevation myocardial infarction (STEMI) before percutaneous coronary intervention (PCI) could affect circulating miR-208b level or not. Methods:Patients diagnosed with STEMI undergoing PCI from January 2016 to July 2017 were enrolled from the Department of Cardiology, Affiliated Zhongshan Hospital of Dalian University.The patients were randomly allocated to two groups: (1) control group (n=25), PCI alone; (2) RIC group (n=50), PCI combined with RIC (three cycles of 5 min inflation and 5 min deflation of the right lower limb with blood pressure cuff performed before reperfusion). Serum miR-208b was measured before and immediately, at 24 h, and 48 h after PCI with real-time quantitative polymerase chain reaction. Results: The expression of miR-208b was significantly higher immediately post PCI than that before operation in the control group (84.1±9.0 vs 77.8±9.4; P=0.032), while it was significantly lower immediately post PCI than that before operationin RIC group (71.0±9.3 vs 77.4±8.8; P=0.028).miR-208b level was similar before PCI between the control and RIC groups (P=0.874), which was significantly reduced immediately post PCI in RIC group as compared with the control group (P=0.021).The peak value of creatine kinase isoenzyme (CK-MB) in the limb RIC group was significantly lower than that in the control group ([135.2±18.6] U/L vs [167.7±17.2] U/L; P=0.038).The area under the CK-MB curve of the RIC group was significantly smaller than that of the control group ([3 060.7±17.1] U/L vs [3 635.9±15.1] U/L); P=0.047]. The left ventricular ejection fraction (LVEF) in RIC group was significantly higher than that in the control group ([57.8±7.8]% vs [51.9±7.9]%; P=0.003) post PCI. The expression level of serum miR-208b was positively correlated with CK-MB AUC in RIC group (r=0.498, P<0.001). Conclusions: RIC of the lower limb prior to PCI could reduce miR-208b level and improve cardiac functionin STEMI patients.

Relationship between total serum IgG concentration and tumor progression in patients with gastric cancer / 国际肿瘤学杂志

Objective To investigate the relationship between total serum IgG concentration and tumor progression in patients with gastric cancer.Methods From January 2010 to May 2017,a total of 90 patients with gastric cancer in Sun Simiao Hospital of Beijing University of Chinese Medicine were enrolled as study subjects and 90 age-and-sex-matched adults from normal physical examination were enrolled as control subjects.The serum IgG subgroup and total IgG concentrations were measured by enzyme-linked immunosorbent assay.The relationships between serum IgG concentration in patients with gastric cancer and clinicopathologic features were analyzed.The numbers of plasma cells with CD138 antibody in both tumor and non-cancerous tissues were evaluated by immunohistochemistry.Results The total serum IgG concentration in patients with gastric cancer was significantly lower than that in healthy control group [(923.6±290.5) mg/dl vs.(1 072.5 ±298.6) mg/dl],with a significant difference (t =3.391,P =0.001).The total serum IgG and IgG1 concentrations were correlated with lymphatic metastasis (t =3.585,P ＜ 0.001;t =3.545,P ＜ 0.001) and gastric cancer stages (t =3.669,P ＜0.001;t =3.401,P ＜0.001).Furthermore,the CD138 + plasma cells in tumor tissues were significantly fewer than those in non-cancerous gastric mucosa (88.7 ± 31.9 vs.108.5 ± 33.4),with significant differences (t =2.778,P =0.007).Multivariate analysis suggested that total serum IgG concentration (HR =0.411,95％ CI:0.094-0.989,P =0.011) and lymph node metastasis (HR =3.148,95％ CI:1.988-5.297,P ＜ 0.001) were independent predictors for poor prognosis.Conclusion Decreased total serum IgG concentration is closely related to tumor progression and poor prognosis in patients with gastric cancer,which indicates that the impaired humoral immunity is associated with tumor progression.