The Ameliorative Effect of Ashwagandha- Withania somnifera (L.) Dunal on park25 Induced Neurodegeneration in Drosophila melanogaster Parkinson’s Disease Model

Ashwagandha - Withania somnifera (L.) Dunal is a perennial shrub belonging to the family Solanaceae. Ashwagandha has been used for over 3000 years in traditional Indian Ayurveda for treatment of various neurological, and stress disorders. The root of Ashwagandha (ASH) is regarded as a tonic, aphrodisiac, narcotic, diuretic, anthelmintic, astringent, thermogenic and stimulant. Ashwagandha with other herbal decoctions was recognized to treat Kampavatha (Parkinson’s Disease) since 18th century. With this wide array of ethnopharmacological relevance, Ashwagandha has been recognized as one of the prominent complementary and alternative medicine to treat many neurodegenerative diseases like Alzheimer’s (AD) and Parkinson’s disease (PD). There is a prominent increase in the cases of AD and PD all over the world and it demands the requirement of complementary and alternative herbal remedies with no/minimal side effects. Many genetic factors are responsible for the onset and progression of PD. Loss-of-function mutations in the parkin gene are a major cause of early onset of autosomal recessive juvenile parkinsonism (AR-JP). Drosophila park25 loss of function mutants exhibit significantly increased number of mitochondria-endoplasmic reticulum contacts and a significantly decreased number of dopaminergic neurons in the adult brain which is the main cause of PD condition. Several studies have demonstrated the ability of Ashwagandha in imparting neuroprotection, improved locomotory ability, memory and learning abilities. The challenge lies in scrutinizing the mechanism and the pathways involved in the neuroprotective properties of this well-known herb. Here in our study, we test the possible neuroprotective effect of Ashwagandha on park25 mutants of Drosophila using lifespan analysis and climbing disability as a disease marker. Parkinson’s mimicking flies were administered with aqueous extraction of Ashwagandha-root mixed with the fly food and subjected to negative geotaxis assay. We observed that there is a prominent increase in the climbing ability in park25 treated flies compared to its age-matched untreated flies. This is the first report showing that, aqueous extraction of Ashwagandha-root extract was able to ameliorate the disease phenotype in the park25 Drosophila Parkinson’s disease model.

Exploration of the Autonomy Issues in Addiction Treatment / 中国医学伦理学

Addiction is a state of chronic neurological dysfunction with repetitive and compulsive behavior as the main manifestation. One of the characteristics of this state is that addicts have significantly reduced autonomy for compulsive behaviors. The emergence of addictive behavior is based on the impaired autonomy of addicts, so the assessment of their condition, prognosis, and related moral responsibility are all related to their autonomy. By exploring whether the treatment of addicts should be based on autonomy as the core indicator to evaluate whether they can end treatment, and exploring the debate on addicts’ autonomy based on the disease model and moral model of addiction, it is believed that whether addicts have autonomy cognition is influenced by the causes of addiction. Furthermore, the issue of moral responsibility of addicts after treatment termination and the quantitative feasibility of autonomy were discussed, and it was advocated to determine the moral responsibility after addiction treatment based on their autonomy.

Establishment and evaluation of a mouse model of schistosomiasis liver disease induced by portal vein injection of worm eggs / 中国热带医学

@#Abstract: Objective　To construct a mouse model of Schistosoma japonicum liver disease induced by direct injection of Schistosoma japonicum eggs through the portal vein and evaluate its effectiveness, in order to provide a new animal model for schistosomiasis liver disease research. Methods　Fifteen 8-week-old C57BL/6 male mice were randomly divided into control group and egg injection group, with 5 in the control group and 10 in the egg injection group. On day -14, 5 000 live eggs were injected into the abdominal cavity of mice, and on day 0, the mice were anesthetized and the abdominal cavity was opened. 5 000 live eggs were injected through the portal vein, and the control group was injected with equal volume of phosphate buffer (PBS). 5 mice in the egg group were killed on day 10 and 30, respectively. The control group mice were killed on day 10, and their serum and liver tissue were collected. Hematoxylin eosin staining (HE) and Masson staining were performed, and alanine aminotransferase (ALT), aspartate aminotransferase (AST), and liver hydroxyproline (HYP) content were detected using a microplate spectrophotometer. Liver fibrosis-related genes, Th1 and Th2 type immune response-related genes were analyzed by real-time fluorescence quantitative PCR (qPCR). Liver injury, egg granuloma and fibrosis, and adaptive immune response were detected to evaluate the effect of portal vein injection of eggs while inducing mouse model of schistosomiasis liver disease. Results　The results showed that significant egg granulomas appeared in the liver of mice after injection of eggs into the portal vein for 10 and 30 days. There was no statistically significant difference in the area of egg granulomas between the 10-day group and the 30-day group (t=0.975, P=0.332). Masson staining and liver hydroxyproline content detection showed significant fibrosis in the liver. The qPCR results showed that, compared with the control group, the expression levels of fibrosis marker genes, such as α⁃Sma (alpha smooth muscle actin), Col1a1 (collagen type Ⅰ alpha 1), and Tgfb1 (transforming growth factor beta 1), were significantly increased (t=6.380, 7.533, 5.314; P=0.002, 0.001, 0.007), and then decreased on the 30th day, with no statistical difference compared to the control group (t=0.940, 1.529, 1.746; P=0.778, 0.543, 0.457). At the same time, the expression levels of Th1 type immune response-related genes, such as tumor necrosis factor alpha (Tnf), interferon gamma (Ifng), and Th2 type immune response-related genes, such as interleukin-5 (Il5), interleukin-13 (Il13), significantly increased 10 days after eggs injection (t=6.163, 4.589, 5.651, 5.367; P=0.003, 0.018, 0.020, 0.009). In addition, there was no significant change in the levels of AST and ALT in the serum of each group of mice (t=0.982, 3.450; P=0.771, 0.074. t=1.164, 0.564; P=0.697, 0.917). Conclusions　A mouse model of schistosomiasis liver disease induced by portal vein injection of worm eggs was constructed. The study provides a new modeling method for studying the mechanism of liver fibrosis in schistosomiasis..

Advances in organoids of the digestive system / 生物工程学报

Organoid is a newly developed cellular there-dimensional culture system in recent years. Organoids have a three-dimensional structure, which is similar to that of the real organs. Together with the characteristics of self-renewal and reproduction of tissue origin, organoids can better simulate the function of real organs. Organoids provide a new platform for the study of organogenesis, regeneration, disease pathogenesis, and drug screening. The digestive system is an essential part of the human body and performs important functions. To date, organoid models of various digestive organs have been successfully established. This review summarizes the latest research progress of organoids of taste buds, esophagi, stomachs, livers and intestines, and prospects future application of organoids.

Base editors: development and applications in biomedicine / 医学前沿

Base editor (BE) is a gene-editing tool developed by combining the CRISPR/Cas system with an individual deaminase, enabling precise single-base substitution in DNA or RNA without generating a DNA double-strand break (DSB) or requiring donor DNA templates in living cells. Base editors offer more precise and secure genome-editing effects than other conventional artificial nuclease systems, such as CRISPR/Cas9, as the DSB induced by Cas9 will cause severe damage to the genome. Thus, base editors have important applications in the field of biomedicine, including gene function investigation, directed protein evolution, genetic lineage tracing, disease modeling, and gene therapy. Since the development of the two main base editors, cytosine base editors (CBEs) and adenine base editors (ABEs), scientists have developed more than 100 optimized base editors with improved editing efficiency, precision, specificity, targeting scope, and capacity to be delivered in vivo, greatly enhancing their application potential in biomedicine. Here, we review the recent development of base editors, summarize their applications in the biomedical field, and discuss future perspectives and challenges for therapeutic applications.

Risk factors of death and prognostic scores of patients with liver cirrhosis undergoing TIPS / 国际生物医学工程杂志

Objective:To analyze the risk factors of death within two years of the patients with liver cirrhosis after transjugular intrahepatic portosystemic shunt(TIPS), and to explore the predictive value of 6 common clinical evaluation systems on the risk of death after TIPS.Methods:TIPS clinical data from 132 patients with liver cirrhosis were analyzed retrospectively. According to the 2-year clinical outcome after TIPS, the patients were divided into the death group and the survival group. Logistic regression was used to analyze the risk factors of death within 2 years after TIPS. According to the scores of CTP, MELD, MELD Na, BioCliM, FIB-4, and ALBI evaluation systems, the prediction efficiency of death risk of the six evaluation systems was evaluated by using the receiver operating characteristic (ROC) curves and the area under the curve (AUC).Results:During the 2-year follow-up period after TIPS, the age, urea nitrogen level, platelet count, and proportion of hepatic encephalopathy in the death group were higher than those in the survival group one month after TIPS, and the serum sodium level was lower than those in the survival group (all P<0.05). Multivariate analysis showed that the elderly and hepatic encephalopathy one month after operation were independent risk factors for death (all P<0.05). At 1 week after the surgery, there were significant differences in CTP, MELD, and MELD-Na scores between the survival group and the death group (all P<0.05). One week after operation, the AUC of ROC of CTP, MELD, MELD-Na, and ALBI scores were 0.685, 0.721, 0.805, and 0.658 respectively, and the optimal critical values were 8.5, 12.99, 14.51 and -1.52 respectively. Conclusions:The elderly and the occurrence of hepatic encephalopathy one month after TIPS are independent risk factors for the death of liver cirrhosis patients after TIPS. The evaluation of CTP, MELD, MELD-Na, and ALBI one week after TIPS can predict the death risk of decompensated liver cirrhosis patients within 2 years after TIPS, and MELD-Na has the best predictive effect.

Islet organoid as a promising model for diabetes

Studies on diabetes have long been hampered by a lack of authentic disease models that, ideally, should be unlimited and able to recapitulate the abnormalities involved in the development, structure, and function of human pancreatic islets under pathological conditions. Stem cell-based islet organoids faithfully recapitulate islet development in vitro and provide large amounts of three-dimensional functional islet biomimetic materials with a morphological structure and cellular composition similar to those of native islets. Thus, islet organoids hold great promise for modeling islet development and function, deciphering the mechanisms underlying the onset of diabetes, providing an in vitro human organ model for infection of viruses such as SARS-CoV-2, and contributing to drug screening and autologous islet transplantation. However, the currently established islet organoids are generally immature compared with native islets, and further efforts should be made to improve the heterogeneity and functionality of islet organoids, making it an authentic and informative disease model for diabetes. Here, we review the advances and challenges in the generation of islet organoids, focusing on human pluripotent stem cell-derived islet organoids, and the potential applications of islet organoids as disease models and regenerative therapies for diabetes.

The evaluation value of three scoring systems for the prognosis of pediatric acute liver failure / 中华实用儿科临床杂志

Objective:To investigate the prognosis value of the Child-Turcotte-Pugh (CTP), pediatrics end-stage liver disease/model for end-stage liver disease(PELD/MELD) and sequential organ failure assessment (SOFA) scores in pediatric acute liver failure (PALF) at 28 th day. Methods:Fifty-four PALF patients admitted in the Pediatric Intensive Care Unit (PICU) and Infection Department of Pediatrics, Qingdao Women′s and Children′s Hospital from June 1, 2012 to June 1, 2019 were included in the study.According to the survival of PALF patients on the 28 th day, they were divided into the survival group (28 cases) and the death group (26 cases). Baseline characte-ristics and laboratory examination data of PALF patients in both groups were collected and compared.Receiver operating characteristic (ROC) curve was used to evaluate the prognostic value of CTP, PELD/MELD and SOFA scores in PALF. Results:The mortality rate of 54 PALF patients was 48.1%.Compared with the survival group, PALF patients in the death group were significantly younger than those in survival group [11.0(3.8-39.0) months vs.14.5(7.3-84.0) months]( Z=-2.145, P=0.020). In addition, CTP, PELD/MELD and SOFA scores were significantly higher in the death group than those in survival group [14.0(11.7-15.0) vs.9.0(7.0-10.0), 32.0(29.0-36.0) vs.25.0(22.0-26.0), 13.0(11.0-16.0) vs.6.0(4.0-7.0)]( Z=-5.095, -4.894, -5.502, all P<0.05). Serum lactate level, blood ammonia level, total bilirubin, direct bilirubin and international normalized ratio were significantly higher in the death group than those in survival group [3.4(2.1-5.3) mmol/L vs.1.5(0.8-2.3) mmol/L, 69.5(46.9-102.9) μmol/L vs.41.7(27.3-50.3) μmol/L, 173.0(97.0-237.2) μmol/L vs.71.9(62.0-136.9) μmol/L, 132.3(53.6-206.2)μmol/L vs.59.3(62.0-99.7) μmol/L, 2.6(1.8-3.5) vs.1.7(1.5-1.9)]( Z=-4.027, -3.220, -2.649, -2.648, -3.807, all P<0.05). Prothrombin time (PT) was significantly prolonged in the death group than that of survival group [27.5(19.2-41.9)s vs.17.8(16.9-22.2)s]( Z=-3.489, P<0.05). Compared with those of survival group, serum albumin, alanine transaminase (ALT) and alpha fetoprotein (AFP) levels were significantly lower in the death group [(30.9±1.0) g/L vs.(33.6±0.9) g/L, 379.2(163.3-880.3) U/L vs.962.5(457.0-1 657.3) U/L, 7.5(0.7-115.8) μg/L vs.22.1(7.9-91.3) μg/L]( t=2.049, Z=-2.510, -2.342, respectively, all P<0.05). The incidence of alimentary tract hemorrhage was significantly higher in the death group than that of survival group (22/26 cases vs.11/28 cases)( χ2=13.340, P<0.05). The cut-off value of CTP, PELD/MELD and SOFA scores in predicting the prognosis of PALF were 11.5, 28.5 and 10.0, respectively.Among the three scoring systems, the specificity and positive predictive value of SOFA scores remained the highest.The sensitivity and specific of a combination of three scoring systems in predicting the prognosis of PALF were 92.3% and 89.3%, respectively, and its Youden index was the highest than that of a single scoring of either CTP, PELD/MELD or SOFA ( Z=2.19, P<0.05). Conclusions:CTP, PELD/MELD and SOFA scores have high predictive value for the short-term prognosis of PALF.The combined detection of the three scoring systems can improve the forecasting efficiency of PALD.

Advances and applications of organoids: a review / 生物工程学报

Novel model systems have provided powerful tools for the research of human biology. Despite of being widely used, the conventional research models could not precisely describe the human physiological phenomenon. Organoids are three-dimensional multicellular aggregates derived from stem cells or organ progenitors that could differentiate and self-organize to recapitulate some specific functionalities and architectures of their in vivo counterpart organs. Organoids can be used to simulate organogenesis because of their human origin. In addition, the genomic stability of organoids could be well maintained during long-term amplification in vitro. Moreover, organoids can be cryopreserved as a live biobank for high-throughput screening. Combinatorial use of organoids with other emerging technologies (e.g. gene editing, organ-on-a-chip and single-cell RNA sequencing) could overcome the bottlenecks of conventional models and provide valuable information for disease modelling, pharmaceutical research, precision medicine and regenerative medicine at the organ level. This review summarizes the classifications, characteristics, current applications, combined use with other technologies and future prospects of organoids.

Protective effects of folic acid on retina in diabetic mice and its anti-oxidative stress mechanism / 中华实验眼科杂志

Objective To explore the protective effects of folic acid on retinas and its anti-oxidative stress mechanism in diabetic mice.Methods Thirty-two 16-week-old SPF degree male db/db mice were randomized into model group and folic acid group,and 16 matched C57BL/KsJ mice were used as controls.Folic acid was used to the mice by oral gavage once per day with the dose of 71 μg/kg (2 ml) for 60 days in the folic acid group,and the same volume of normal saline solution was used in the model group and control group in the same way.The activities,mental state,body weight,and fasting plasma glucose (FPG) of the mice were recorded during experiment.At the end of the intervention,the mice were sacrificed and the retinas and blood sample were obtained.The histopathology of the retinas was examined with hematoxylin-eosin staining;serum homocysteine (Hcy) was detected by ELISA assay;the relative expressions of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were detected in the retinas by real-time fluorescence quantitative PCR;the relative expressions of B lymphoma 2 protein (bcl-2),bcl-2 related X protein (bax),3-nitrotyrosine (3-NT) and 4-Hydroxynonine (4-HNE) proteins were assayed by Western blot assay;superoxide dismutase (SOD),8-hydroxydeoxyguanosine (8-OHdG) and malondialdehyde (MDA) levels in the retinas were detected by biochemical kits,and immunofluorescence assay was used to detect the expression of NADPH oxidation 4 (NOX4) in the retinas.The use and care of the experimental animals adhered to the ARVO Statement by the American Association for Vision and Ophthalmology Research and this study protocol was approved by Ethic Committee of Qinghai University (QHDX 2018-35).Results Over the experimental period,The FPG was normal and body weight was gradually increased in the mice of control group.The FPG＞16.7 mmol/L and the mice appeared obese.In the folic acid group,both body mass and FPG of the mice were gradually reduced.At the end of drug administration,serum Hcy concentration of the mice was (27.18± 3.18)μmol/L in the model group,which was significantly higher than (8.28±2.18) μmol/L in the control group and (13.73±2.54) μmol/L in the folic acid group (all at P＜0.05).The retinal structure was intact in the control group,and the retinas were thinning with more capillaries and inflammatory cells in the model group,the thickness of the retinas was increased and the capillaries and inflammatory cells were decreased in the folic acid group.The relative expressions of TNF-α and IL-6 mRNA in the retinas were significantly higher in the model group than those in the control group,and those in the folic acid group were reduced in comparison with the model group (all at P＜0.05).The relative expression of bcl-2 protein in the retinas of folic acid group was lower than that in the control group and higher than that in the model group,and the relative expressions of bax,3-NT and 4-HNE proteins in the retinas of the folic acid group were significantly higher than those in the control group and lower than those in the model group (all at P＜0.05).The T-SOD activity in the folic acid group was significantly stronger than that in the control group and weaker than that in the model group,and the concentrations of 8-OHdG and MDA in the retinas of the folic acid group were significantly reduced in comparison with those of the control group and elevated in comparison with those of the model group (all at P＜0.05).The expressing intensity of NOX4 protein in the retinas of the folic acid group was significantly weaker than that of the model group.Conclusions Folic acid appears aprotective effect on retinal tissue in diabetic mice by reducing serum Hcy,inhibiting oxidative stress and cell apoptosis.

Induction effect of DNCB on atopic dermatitis BALB/c mice and its mechanism / 吉林大学学报(医学版)

Objective: To induce the atopic dermatitis (AD) by 2, 4-dinitrobenzene (DNCB) in the BALB/c mice, and to explore the possible mechanism of DNCB as hapten to induce AD. Methods: A total of 12 BALB/c mice were randomly divided into control group (n = 6) and AD model group (n = 6). The dorsal skin of the mice in AD model group was sensitized by 1.0% DNCB at days 1, 4, and 7 and the back skin of the left ears were challenged by 0. 5% DNCB at days 14, 17, 19, 22, 24, 27, and 29. The mice in control group were given substrate solution with the same volume at the same time points. The inflammation scores of the skin tissue of the mice in two groups were evaluated, the appearance changes of the ear skin of the mice in two groups were observed, the pathomorphology of the skin tissue in left ear of the mice in two groups were observed by HE staining and toluidine blue staining, the epidermal thickness of the skin tissue at lesion site of the mice in two groups was measured, and the number of mast cells in the skin tissue at lesion site of the mice in two groups was counted; the expression levels of interleukin-4 (IL-4) in the skin tissue at lesion site of the mice in two groups was detected by immunohistochemistry staining, and the levels of serum IgE was detected by ELISA method. Results: The inflammation score of the mice in AD model group was higher that in control group (P < 0. 01). The skin tissue in left ear of the mice in AD model presented as redness swelling and hardening accompaning with abnormal scratching marks; the epidermal thickness, the number of mast cells, the expression level of IL-4 in skin tissue at lesion site and the level of serum IgE of the mice in AD model group were all increased compared with control group (P < 0. 05 or P < 0. 01). Conclusion: The AD mouse can be successfully induced by 1. 0% DNCB for sensitizing and 0. 5% DNCB for chllenging in the BALB/c mice, and its mechanism may be related with the increasing of IL-4 expression level in skin tissue and IgE level in serum.

Protective effects of folic acid on retina in diabetic mice and its anti-oxidative stress mechanism / 中华实验眼科杂志

Objective@#To explore the protective effects of folic acid on retinas and its anti-oxidative stress mechanism in diabetic mice.@*Methods@#Thirty-two 16-week-old SPF degree male db/db mice were randomized into model group and folic acid group, and 16 matched C57BL/KsJ mice were used as controls.Folic acid was used to the mice by oral gavage once per day with the dose of 71 μg/kg (2 ml) for 60 days in the folic acid group, and the same volume of normal saline solution was used in the model group and control group in the same way.The activities, mental state, body weight, and fasting plasma glucose (FPG) of the mice were recorded during experiment.At the end of the intervention, the mice were sacrificed and the retinas and blood sample were obtained.The histopathology of the retinas was examined with hematoxylin- eosin staining; serum homocysteine (Hcy) was detected by ELISA assay; the relative expressions of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were detected in the retinas by real-time fluorescence quantitative PCR; the relative expressions of B lymphoma 2 protein (bcl-2), bcl-2 related X protein (bax), 3-nitrotyrosine (3-NT) and 4-Hydroxynonine (4-HNE) proteins were assayed by Western blot assay; superoxide dismutase (SOD), 8-hydroxydeoxyguanosine (8-OHdG) and malondialdehyde (MDA) levels in the retinas were detected by biochemical kits, and immunofluorescence assay was used to detect the expression of NADPH oxidation 4 (NOX4) in the retinas.The use and care of the experimental animals adhered to the ARVO Statement by the American Association for Vision and Ophthalmology Research and this study protocol was approved by Ethic Committee of Qinghai University (QHDX 2018-35).@*Results@#Over the experimental period, The FPG was normal and body weight was gradually increased in the mice of control group.The FPG>16.7 mmol/L and the mice appeared obese.In the folic acid group, both body mass and FPG of the mice were gradually reduced.At the end of drug administration, serum Hcy concentration of the mice was (27.18±3.18)μmol/L in the model group, which was significantly higher than (8.28±2.18)μmol/L in the control group and (13.73±2.54)μmol/L in the folic acid group (all at P<0.05). The retinal structure was intact in the control group, and the retinas were thinning with more capillaries and inflammatory cells in the model group, the thickness of the retinas was increased and the capillaries and inflammatory cells were decreased in the folic acid group.The relative expressions of TNF-α and IL-6 mRNA in the retinas were significantly higher in the model group than those in the control group, and those in the folic acid group were reduced in comparison with the model group (all at P<0.05). The relative expression of bcl-2 protein in the retinas of folic acid group was lower than that in the control group and higher than that in the model group, and the relative expressions of bax, 3-NT and 4-HNE proteins in the retinas of the folic acid group were significantly higher than those in the control group and lower than those in the model group (all at P<0.05 ). The T-SOD activity in the folic acid group was significantly stronger than that in the control group and weaker than that in the model group, and the concentrations of 8-OHdG and MDA in the retinas of the folic acid group were significantly reduced in comparison with those of the control group and elevated in comparison with those of the model group (all at P<0.05). The expressing intensity of NOX4 protein in the retinas of the folic acid group was significantly weaker than that of the model group.@*Conclusions@#Folic acid appears aprotective effect on retinal tissue in diabetic mice by reducing serum Hcy, inhibiting oxidative stress and cell apoptosis.

Structural changes of retinal microcirculation in mouse model of ischemia-reperfusion injury / 中华实验眼科杂志

Objective To study the structural changes of retinal microcirculation in mouse model of ischemia reperfusion injury(RIR).Methods Ninty healthy male C57BL/6 mice were devided into normal group and RIR group by using random number table method,45 mice for each group.The left eyes in both groups were used in this study.The mouse model of RIR was established by perfusion of saline solution in the anterior chamber.The damage of microcirculation was analyzed from different levels of vascular structure and function by retinal blood vessel staining,FITC angiography,histopathological examination,and ultrostructure of retinal vessels was observed with transmission electron microscope.The experimental animals and experimental conditions complied with the Regulations on the Management of Laboratory Animals of Kunming Medical University.Results There was no significant difference in retinal arterioles diameter in the RIR group compared to the normal control group (P=0.350).The small veins were obviously dilated in comparison with normal control group (P =0.03) and were in hyper-congestive state.The artery/vein value was 0.76±0.03 in the RIR group,and that in the normal control group was 0.97±0.01,with significant difference between them (P=0.000).The main changes were capillary occlusion,non-perfusion zone formation and other structural changes,accompaning by blood-retinal barrier damage;the destruction of capillary endothelial cells and pericytes and basement membrane thickening were seen under the transmission electron microscope.Conclusions RIR mainly results in capillary damage,including capillary barrier damage,capillary occlusion and non-perfusion area formation.

Improvement Effects of Shenrong Bunao Capsule on Learning and Memory Ability of Alzheimer ’s Disease Model Mice and Its Mechanism Study / 中国药房

OBJECTIVE： To study the improvement effect of Shenrong bunao capsule on learning and memory ability of Alzheimer’s disease （AD） model mice， and to investigate its mechanism. METHODS： Totally 72 mice were randomly divided into blank control group （normal saline）， model group （normal saline）， Piracetam tablets group （positive control，0.80 g/kg），Shenrong bunao capsule high-dose，middle-dose and low-dose groups （1.92， 0.96， 0.48 g/kg）， with 12 mice in each group. Except that blank control group was given constant volume of normal saline subcutaneously. Other groups were given D-galactose （150 mg/kg） subcutaneously and sodium nitrite （50 mg/kg） intraperitoneally every day to induce AD model. At the same time，they were given relevant medicine intragastrically，once a day， for consecutive 60 d. 1 h after last administration， Morris water maze test was used to measure escape latency and times of crossing the platform within 90 s. HE staining was used to observe pathological changes of cerebral cortex in mice. Immunohistochemistry was used to detect the expressions of TNF-α， NF-κB p65， PI3K and Akt in cerebral cortex of mice. RESULTS： Compared with blank control group， escape latency was prolonged significantly （P＜0.01）， and the times of crossing the platform within 90 s was decreased significantly in model group （P＜0.01）. The neurons in cerebral cortex was damaged obviously， and the number of intact neurons was decreased significantly （P＜0.01）. The protein expressions of TNF-α， NF-κB p65， PI3K and Akt in cerebral cortex were increased significantly （P＜0.01）. Compared with model group， except that there was no statistical significance in escape latency， protein expressions of TNF-α， NF-кB p65， PI3K and Akt in Shenrong bunao capsule low-dose group （P＞0.05）， above indexes of other administration groups were improved significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： Shenrong bunao capsule can improve the learning and memory ability of AD model mice， and its mechanism may be related to the inhibiting the protein expressions of TNF-α，NF-κB p65， PI3K and Akt in cerebral cortex region and relieving inflammation injury so as to protect cranial nerve.

Research progress of Cre/LoxP system in the eyes of animal model / 中华实验眼科杂志

Cre/LoxP system has been widely used as new type of gene targeting,which can transform genes in specific tissues and organs,remove specific gene fragments,and help to study the effects of specific genes on growth and development. Cre/LoxP system is the core technology of conditional gene targeting,which induces gene targeting, and spatiotemporal specific gene targeting. The resource about the mechanism of the Cre/LoxP system is helpful to obtain the Cre transgenic mouse model on different cell types of the retina. The Cre transgenic mice with specific expression in retinal bipolar cells and ganglion cells can provide the experimental basis for the absence of specific tissue single genes during growth and development. This review focused on the Cre/LoxP system and specific Cre recombinase mouse model in eyes.

Neurological Disorder Brain Model: A Lesson from Marine Worms (Annelida: Polychaeta)

@#The incidence of neurodegenerative diseases is directly proportional to age. The prevalence of non-communicable diseases, for example, Alzheimer’s and Parkinson’s, is expected to rise in the coming years. Understanding the etiopathology of these diseases is a crucial step that needs to be taken to develop drugs for their treatment. Animal models are being increasingly used to expand the knowledge and understanding on neurodegenerative diseases. Marine worms, known as polychaetes (phylum Annelida), which are abundantly and frequently found in benthic environments, possess a simple yet complete nervous system (including a true brain that is centralised and specialised) compared to other annelids. Hence, polychaetes can potentially be the next candidate for a nerve disease model. The ability to activate the entire nervous system regeneration (NSR) is among the remarkable features of many polychaetes species. However, the information on NSR in polychaetes and how it can potentially model neurodegenerative diseases in humans is still lacking. By exploring such studies, we may eventually be able to circumvent the developmental constraints that limit NSR in the human nervous system. This article is intended to briefly review responsible mechanisms and signalling pathways of NSR in marine polychaetes and to make a comparison with other established models of neurodegenerative disease.

Establishment and evaluation of rat models of myocardial ischemia reperfusion-related no-reflow under non-artificial ventilator / 吉林大学学报(医学版)

Objective: To establish the rat models of myocardial ischemia reperfusion-related no-reflow under non-artificial ventilator, to evaluate the models by morphology, hematological biochemistry and hemorheology, and to lay the foundation for studying the pathogenesis of cardiovascular diseases and evaluating the pharmacodynamics of drugs of cardiovascular diseases. Methods: Fifty healthy female Wistar rats were randomly divided into sham operation group (n=20) and model group (n=30). The myocardial ischemia reperfusion-related no-reflow rat models in model group were induced by ligation of left anterior descending coronary artery for 2 h and another 2 h for reperfusion. The rats in sham group were only threaded and not ligated. The model was evaluated by detecting the area at risk (AAR), area at infarct (AAD, area at no-reflow (AAN) of myocardium, the activities of creatine kinase MB (CK-MB), lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) in serum, the whole blood viscosity, the plasma viscosity, the platelet adhesion rate (PAR), the platelet aggregation (PAG) of the rats. Results: Compared with sham operation group, the AAR, AAI, and AAN of myocardium, the activities of CK-MB, LDH, and AST in serum, the whole blood low shear viscosity (20/s), the middle shear viscosity (60/s), the high shear viscosity (120/s), the plasma viscosity, PAR, PAG (1, 3, 5 min) and the maximum platelet aggregation rate (MAPG) of the rats in model group were significantly increased (P<0. 05 or P<0. 01). Conclusion: The rat model of myocardial ischemia reperfusion-related no-reflow under non-artificial ventilator is successfully established. This method is characterized by simple operation, decreased injury to animals, higher stability of model construction, shorter experimental cycle and less cost.

Application of in vitro porcine gastric model of submucosal eminence lesion in endoscopic submucosal dissection / 中华消化内镜杂志

Objective@#To establish the in vitro porcine gastric model of submucosal eminence lesion and to evaluate its application to endoscopic submucosal dissection(ESD).@*Methods@#Silicone rubber impression materials and steel balls with diameters of 1 cm, 2 cm, and 3 cm were used to make three pairs of spherical cavities. And then raw ground beef was put into spherical cavities and boiled for 20 minutes to make spherical mass models. Six isolated porcine stomach with esophagus and duodenum were selected. The mass models with diameters of 1 cm, 2 cm and 3 cm were imbedded respectively into the submucosa of fundus, body, and antrum of porcine stomach through the incision on serosal layer. The submucosal masses were observed by endoscopy and endoscopic ultrasonography and ESD was performed.@*Results@#A total of 18 mass models were constructed in 6 porcine stomachs, of which 17 models were successfully established and 1 failed. Typical endoscopic characteristics of gastric submucosal eminence lesions were found in 17 models. Endoscopic ultrasonography showed that these models originated from submucosal layer and demonstrated mixed echo. There were no significant differences between mucosa of lesions and that of surrounding areas. ESD was successfully performed in the porcine gastric models of submucosal eminence lesions, and all models were not broken or detached.@*Conclusion@#The in vitro porcine gastric model of submucosal eminence lesions can well replicate disease status and provide a suitable model for study on endoscopic therapy of submucosal eminence lesion and training of endoscopists.

Research progress of Cre/LoxP system in the eyes of animal model / 中华实验眼科杂志

Cre/LoxP system has been widely used as new type of gene targeting, which can transform genes in specific tissues and organs, remove specific gene fragments, and help to study the effects of specific genes on growth and development.Cre/LoxP system is the core technology of conditional gene targeting, which induces gene targeting, and spatiotemporal specific gene targeting.The resource about the mechanism of the Cre/LoxP system is helpful to obtain the Cre transgenic mouse model on different cell types of the retina.The Cre transgenic mice with specific expression in retinal bipolar cells and ganglion cells can provide the experimental basis for the absence of specific tissue single genes during growth and development.This review focused on the Cre/LoxP system and specific Cre recombinase mouse model in eyes.

Efeito do fotossensibilizador butyl azul de toluidina na terapia fotodinâmica antimicrobiana para o tratamento da periodontite experimental em ratos / Effects of butyl toluidine blue photosensitizer on antimicrobial photodynamic therapy for experimental periodontitis treatment in rats

O presente estudo avaliou pela primeira vez "in vivo" os efeitos de três concentrações do butyl azul de toluidina (BuTB) como agente fotossensibilizador na terapia fotodinâmica antimicrobiana (aPDT), como terapia coadjuvante a raspagem e alisamento radicular (RAR), para o tratamento de periodontite experimental (PE) em ratos. A PE foi induzida por meio da instalação de um fio de algodão ao redor do primeiro molar inferior esquerdo. Posteriormente os animais foram aleatoriamente distribuídos em 7 grupos com 15 animais cada, através de uma tabela gerada por computador, de acordo com os seguintes tratamentos: RAR (n=15) - RAR seguido de irrigação local de solução salina fisiológica; BuTB-0,1 (n=15) - RAR seguido de aplicação local de BuTB na concentração de 0,1 mg/mL; aPDT-0,1 (n=15) - RAR seguido da aplicação local de BuTB na concentração de 0,1 mg/mL e irradiação com laser de diodo (LD) de InGaAlP (660 nm, 40 mW, 60 s, 2,4 J); BuTB-0,5 (n=15) ­ RAR seguido de aplicação local de BuTB na concentração de 0,5 mg/mL; aPDT-0,5 (n=15) ­ RAR seguido da aplicação local de BuTB na concentração de 0,5 mg/mL e irradiação com LD; BuTB-2,0 (n=15) - RAR seguido de aplicação local de BuTB na concentração de 2 mg/mL; aPDT-2,0 (n=15) - RAR seguido da aplicação local de BuTB na concentração de 2 mg/mL e irradiação com LD. Decorridos 7, 15 e 30 dias pós-tratamento, 5 animais de cada grupo foram submetidos à eutanásia. A área de furca dos molares foi submetida às análises histológica, histométrica e dos padrões de imunomarcação para TGF-ß1, OCN e TRAP. Os dados foram submetidos à análise estatística (α = 5%). De acordo com a análise histométrica na região de furca, todos os grupos experimentais apresentaram menor perda óssea comparado ao grupo controle. Histologicamente, os espécimes do aPDT-0,5 apresentaram uma resposta inflamatória local mais branda e menos extensa, com melhor reestruturação tecidual em todos os períodos. Aos 30 dias observou-se resolução total da resposta inflamatória local, com presença de tecido conjuntivo denso. Alguns espécimes apresentavam trabéculas ósseas com contorno regular revestido com osteoblastos ativos, incluindo áreas de neoformação óssea. O tratamento com aPDT na concentração de 0,5 mg/mL resultou em padrões mais altos de imunomarcação de TGF-ß1 em todos os períodos e de OCN aos 30 dias. Diante dos resultados obtidos, todas as concentrações do novo fotossensibilizador BuTB trouxeram resultados adicionais ao tratamento da PE em relação a RAR. No entanto, a aPDT realizada com a concentração de 0,5 mg/mL resultou em benefícios adicionais na resposta inflamatória local e melhor reestruturação tecidual(AU)

The present study evaluated for the first time the effects of three concentrations of butyl toluidine blue (BuTB) as a photosensitizing agent on antimicrobial photodynamic therapy (aPDT), as adjuvant therapy to scaling and root planing (SRP), for the treatment of experimental periodontitis (EP) in rats. EP was induced by placing a cotton thread around the lower left first molar. Subsequently, the animals were randomly distributed into seven groups with 15 animals each, through a computer generated table, according to the following treatments: SRP (n = 15), SRP followed by local irrigation of physiological saline solution; BuTB-0.1 (n = 15), SRP followed by local application of 0.1 mg/mL BuTB; aPDT-0.1 (n = 15), SRP followed by local application of BuTB at 0.1 mg/mL concentration and irradiation with InGaAlP diode laser (DL) (660 nm, 40 mW, 60 s, 4 J); BuTB-0.5 (n = 15), SRP followed by local application of BuTB at 0.5 mg/mL concentration; aPDT-0.5 (n = 15), SRP followed by local application of BuTB at 0.5 mg/mL concentration and DL irradiation; BuTB-2.0 (n = 15), SRP followed by local application of BuTB at 2 mg/mL concentration; aPDT-2.0 (n = 15), SRP followed by local application of BuTB at 2 mg/mL concentration and DL irradiation. The animals (n=5) from each group were submitted to euthanasia at 7, 15 and 30 days post-treatment. The furcation area of the first lower molar was submitted to histological, histometric and immunohistochemical analyses to identify TGF-ß1, OCN and TRAP. The data were submitted to statistical analysis (α = 5%). According to the histometric analysis in the furcation region, all experimental groups presented lower bone loss compared to the control group. Histologically, the aPDT -0.5 specimens presented a milder and less extensive local inflammatory response, with better tissue remodeling in all periods. Total resolution of the local inflammatory response was observed at 30 days with presence of mature connective tissue. Some specimens presented bone trabeculae with a regular contour and active osteoblasts, including areas of bone neoformation. Treatment with aPDT-0.5 also resulted in higher immunolabelling patterns of TGFß1 at all periods and of OCN at 30 days. All concentrations of the new photosensitizer BuTB resulted in significant improvement for EP treatment in relation to SRP. However, aPDT combined with BuTB at 0.5 mg / mL showed the best benefits for inflammatory response and periodontal repair process(AU)