Developments in the study of Chinese herbal medicine's assessment index and action mechanism for diabetes mellitus

In traditional Chinese medicine(TCM),based on various pathogenic symptoms and the‘golden chamber’medical text,Huangdi Neijing,diabetes mellitus falls under the category‘collateral disease’.TCM,with its wealth of experience,has been treating diabetes for over two millennia.Different antidiabetic Chinese herbal medicines re-duce blood sugar,with their effective ingredients exerting unique advantages.As well as a glucose lowering effect,TCM also regulates bodily functions to prevent diabetes associated complications,with reduced side effects compared to western synthetic drugs.Chinese herbal medicine is usually composed of polysaccharides,saponins,al-kaloids,flavonoids,and terpenoids.These active ingredients reduce blood sugar via various mechanism of actions that include boosting endogenous insulin secretion,enhancing insulin sensitivity and adjusting key enzyme activity and scavenging free radicals.These actions regulate glycolipid metabolism in the body,eventually achiev-ing the goal of normalizing blood glucose.Using different animal models,a number of molecular markers are available for the detection of diabetes induction and the molecular pathology of the disease is becoming clearer.Nonetheless,there is a dearth of scientific data about the pharmacology,dose-effect relationship,and structure-activity relationship of TCM and its constituents.Further research into the efficacy,toxicity and mode of action of TCM,using different metabolic and molecular markers,is key to developing novel TCM antidiabetic formulations.

Expansion of human umbilical cord derived mesenchymal stem cells in regenerative medicine

BACKGROUND Stem cells are undifferentiated cells that possess the potential for self-renewal with the capacity to differentiate into multiple lineages.In humans,their limited numbers pose a challenge in fulfilling the necessary demands for the regeneration and repair of damaged tissues or organs.Studies suggested that mesenchymal stem cells(MSCs),necessary for repair and regeneration via transplantation,require doses ranging from 10 to 400 million cells.Furthermore,the limited expansion of MSCs restricts their therapeutic application.AIM To optimize a novel protocol to achieve qualitative and quantitative expansion of MSCs to reach the targeted number of cells for cellular transplantation and minimize the limitations in stem cell therapy protocols.METHODS Human umbilical cord(hUC)tissue derived MSCs were obtained and re-cultured.These cultured cells were subjected to the following evaluation pro-cedures:Immunophenotyping,immunocytochemical staining,trilineage differentiation,population doubling time and number,gene expression markers for proliferation,cell cycle progression,senescence-associatedβ-galactosidase assay,human telomerase reverse transcriptase(hTERT)expression,mycoplasma,cytomegalovirus and endotoxin detection.RESULTS Analysis of pluripotent gene markers Oct4,Sox2,and Nanog in recultured hUC-MSC revealed no significant differences.The immunophenotypic markers CD90,CD73,CD105,CD44,vimentin,CD29,Stro-1,and Lin28 were positively expressed by these recultured expanded MSCs,and were found negative for CD34,CD11b,CD19,CD45,and HLA-DR.The recultured hUC-MSC population continued to expand through passage 15.Proliferative gene expression of Pax6,BMP2,and TGFb1 showed no significant variation between recultured hUC-MSC groups.Nevertheless,a significant increase(P<0.001)in the mitotic phase of the cell cycle was observed in recultured hUC-MSCs.Cellular senescence markers(hTERT expression andβ-galactosidase activity)did not show any negative effect on recultured hUC-MSCs.Additionally,quality control assessments consistently confirmed the absence of mycoplasma,cytomegalovirus,and endotoxin contamination.CONCLUSION This study proposes the development of a novel protocol for efficiently expanding stem cell population.This would address the growing demand for larger stem cell doses needed for cellular transplantation and will significantly improve the feasibility of stem cell based therapies.

Transcription regulators differentiate mesenchymal stem cells into chondroprogenitors,and their in vivo implantation regenerated the intervertebral disc degeneration

BACKGROUND Intervertebral disc degeneration(IVDD)is the leading cause of lower back pain.Disc degeneration is characterized by reduced cellularity and decreased production of extracellular matrix(ECM).Mesenchymal stem cells(MSCs)have been envisioned as a promising treatment for degenerative illnesses.Cell-based therapy using ECM-producing chondrogenic derivatives of MSCs has the potential to restore the functionality of the intervertebral disc(IVD).AIM To investigate the potential of chondrogenic transcription factors to promote differentiation of human umbilical cord MSCs into chondrocytes,and to assess their therapeutic potential in IVD regeneration.METHODS MSCs were isolated and characterized morphologically and immunologically by the expression of specific markers.MSCs were then transfected with Sox-9 and Six-1 transcription factors to direct differentiation and were assessed for chondrogenic lineage based on the expression of specific markers.These differentiated MSCs were implanted in the rat model of IVDD.The regenerative potential of transplanted cells was investigated using histochemical and molecular analyses of IVDs.RESULTS Isolated cells showed fibroblast-like morphology and expressed CD105,CD90,CD73,CD29,and Vimentin but not CD45 antigens.Overexpression of Sox-9 and Six-1 greatly enhanced the gene expression of transforming growth factor beta-1 gene,BMP,Sox-9,Six-1,and Aggrecan,and protein expression of Sox-9 and Six-1.The implanted cells integrated,survived,and homed in the degenerated intervertebral disc.Histological grading showed that the transfected MSCs regenerated the IVD and restored normal architecture.CONCLUSION Genetically modified MSCs accelerate cartilage regeneration,providing a unique opportunity and impetus for stem cell-based therapeutic approach for degenerative disc diseases.

Combination of Shengmai San and Radix puerariae ameliorates depression-like symptoms in diabetic rats at the nexus of PI3K/BDNF/SYN protein expression

Background:Hyperglycemia is a characteristic feature of diabetes that often results in neuropsychological complications such as depression.Diabetic individuals are more vulnerable to experience depression compared to the normal population.Thus,novel treatment approaches are required to reduce depressive symptoms among diabetic individuals.Traditional Chinese medicines(TCMs)such as Shengmai San(SMS)and Radix puerariae(R)are usually widely used to treat ailments such as neurological com-plications since ancient time.Methods:In this study,SMS was combined with R to prepare an R-SMS formulation and screened for their antidepressant activity in diabetic rats.The antidepressant po-tential of the prepared combination was evaluated behaviorally using open field test,novelty-induced hypophagia,and forced swim test in diabetic rats with biochemical and protein expression(PI3K,BDNF[brain-derived neurotrophic factor],and SYN[pr-esynaptic vesicle protein])analysis.Results:Diabetic rats(streptozotocin,45 mg/kg)showed elevated fasting blood glu-cose(FBG)>12 mM with depressive symptoms throughout the study.Treatment with R-SMS(0.5,1.5,and 4.5 g/kg)significantly reverted depressive symptoms in diabetic rats as evinced by significantly(p<0.05)reduced immobility time with an increased tendency to eat food in a novel environment.Treatment with R-SMS also significantly increased the protein expression of PI3K,BDNF,and SYN protein,which play a crucial role in depression.Conclusion:This study showed that R-SMS formulation antagonized depressive symptoms in diabetic rats;thus,this formulation might be studied further to develop as an antidepressant.

Zinc enhances the cell adhesion,migration,and self-renewal potential of human umbilical cord derived mesenchymal stem cells

BACKGROUND Zinc(Zn)is the second most abundant trace element after Fe,present in the human body.It is frequently reported in association with cell growth and proliferation,and its deficiency is considered to be a major disease contributing factor.AIM To determine the effect of Zn on in vitro growth and proliferation of human umbilical cord(hUC)-derived mesenchymal stem cells(MSCs).METHODS hUC-MSCs were isolated from human umbilical cord tissue and characterized based on immunocytochemistry,immunophenotyping,and tri-lineage differentiation.The impact of Zn on cytotoxicity and proliferation was determined by MTT and Alamar blue assay.To determine the effect of Zn on population doubling time(PDT),hUC-MSCs were cultured in media with and without Zn for several passages.An in vitro scratch assay was performed to analyze the effect of Zn on the wound healing and migration capability of hUC-MSCs.A cell adhesion assay was used to test the surface adhesiveness of hUC-MSCs.Transcriptional analysis of genes involved in the cell cycle,proliferation,migration,and selfrenewal of hUC-MSCs was performed by quantitative real-time polymerase chain reaction.The protein expression of Lin28,a pluripotency marker,was analyzed by immunocytochemistry.RESULTS Zn at lower concentrations enhanced the rate of proliferation but at higher concentrations(>100μM),showed concentration dependent cytotoxicity in hUC-MSCs.hUC-MSCs treated with Zn exhibited a significantly greater healing and migration rate compared to untreated cells.Zn also increased the cell adhesion rate,and colony forming efficiency(CFE).In addition,Zn upregulated the expression of genes involved in the cell cycle(CDC20,CDK1,CCNA2,CDCA2),proliferation(transforming growth factorβ1,GDF5,hypoxia-inducible factor 1α),migration(CXCR4,VCAM1,VEGF-A),and self-renewal(OCT4,SOX2,NANOG)of hUC-MSCs.Expression of Lin28 protein was significantly increased in cells treated with Zn.CONCLUSION Our findings suggest that zinc enhances the proliferation rate of hUC-MSCs decreasing the PDT,and maintaining the CFE.Zn also enhances the cell adhesion,migration,and self-renewal of hUC-MSCs.These results highlight the essential role of Zn in cell growth and development.

Wnt signaling pathway inhibitor promotes mesenchymal stem cells differentiation into cardiac progenitor cells in vitro and improves cardiomyopathy in vivo

BACKGROUND Cardiovascular diseases particularly myocardial infarction(MI)are the leading cause of mortality and morbidity around the globe.As cardiac tissue possesses very limited regeneration potential,therefore use of a potent small molecule,inhibitor Wnt production-4(IWP-4)for stem cell differentiation into cardiomyocytes could be a promising approach for cardiac regeneration.Wnt pathway inhibitors may help stem cells in their fate determination towards cardiomyogenic lineage and provide better homing and survival of cells in vivo.Mesenchymal stem cells(MSCs)derived from the human umbilical cord have the potential to regenerate cardiac tissue,as they are easy to isolate and possess multilineage differentiation capability.IWP-4 may promote the differentiation of MSCs into the cardiac lineage.AIM To evaluate the cardiac differentiation ability of IWP-4 and its subsequent in vivo effects.METHODS Umbilical cord tissue of human origin was utilized to isolate the MSCs which were characterized by their morphology,immunophenotyping of surface markers specific to MSCs,as well as by tri-lineage differentiation capability.Cytotoxicity analysis was performed to identify the optimal concentration of IWP-4.MSCs were treated with 5μM IWP-4 at two different time intervals.Differentiation of MSCs into cardiomyocytes was evaluated at DNA and protein levels.The MI ratmodel was developed.IWP-4 treated as well as untreated MSCs were implanted in the MI model,then the cardiac function was analyzed via echocardiography.MSCs were labeled with 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate(DiI)dye for tracking,while the regeneration of infarcted myocardium was examined by histology and immunohistochemistry.RESULTS MSCs were isolated and characterized.Cytotoxicity analysis showed that IWP-4 was non-cytotoxic at 5μM concentration.Cardiac specific gene and protein expression analyses exhibited more remarkable results in fourteen days treated group that was eventually selected for in vivo transplantation.Cardiac function was restored in the IWP-4 treated group in comparison to the MI group.Immunohistochemical analysis confirmed the homing of pre-differentiated MSCs that were labeled with DiI cell labeling dye.Histological analysis confirmed the significant reduction in fibrotic area,and improved left ventricular wall thickness in IWP-4 treated MSC group.CONCLUSION Treatment of MSCs with IWP-4 inhibits Wnt pathway and promotes cardiac differentiation.These pre-conditioned MSCs transplanted in vivo improved cardiac function by cell homing,survival,and differentiation at the infarcted region,increased left ventricular wall thickness,and reduced infarct size.

Antiglycation,antioxidant and toxicological potential of polyphenol extracts of alligator pepper,ginger and nutmeg from Nigeria

Objective:To evaluate the antioxidant and antiglycation potential of polyphenols from three spices;alligator pepper,ginger and nutmeg.Methods:Polyphenol extracts of these spices were subjected to brine-shrimp lethality assay,phyloloxicily test,DPPH and superoxide anion radical scavenging as well as BSA-glucose antiglycation assay.Results:Results obtained showed that polyphenol extract of ginger has the highest anlioxidant potential with IC_(50)0.075 and 0.070 mg/mL.for DPPH and superoxide anion radical scavenging assay while alligator pepper displayed highest antiglycation activity with IC_(50)0.125 mg/mL.However,nutmeg extract exhibited weakest cytotoxic and phytotoxic potential with LD_(50)4359.70 and 1490μg/mL respectively.Conclusions:It can be concluded that the polyphenol extracts of alligator pepper,ginger and nutmeg displayed good antioxidant as well as antiglycation potential and are,safe for consumption.

Effect of glycyrrhizic acid and 18β-glycyrrhetinic acid on the differentiation of human umbilical cord-mesenchymal stem cells into hepatocytes

BACKGROUND End-stage liver disease is a global health complication with high prevalence and limited treatment options.Cell-based therapies using mesenchymal stem cells(MSCs)emerged as an alternative approach to support hepatic regeneration.In vitro preconditioning strategies have been employed to strengthen the regenerative and differentiation potential of MSCs towards hepatic lineage.Chemical compounds of the triterpene class;glycyrrhizic acid(GA)and 18β-glycyrrhetinic acid(GT)possess diverse therapeutic properties including hepatoprotection and anti-fibrosis characteristics.They are capable of modulating several signaling pathways that are crucial in hepatic regeneration.Preconditioning with hepato-protective triterpenes may stimulate MSC fate transition towards hepatocytes.AIM To explore the effect of GA and GT on hepatic differentiation of human umbilical cord-MSCs(hUC-MSCs).METHODS hUC-MSCs were isolated and characterized phenotypically by flow cytometry and immunocytochemistry for the expression of MSC-associated surface molecules.Isolated cells were treated with GA,GT,and their combination for 24 h and then analyzed at three time points;day 7,14,and 21.qRT-PCR was performed for the expression of hepatic genes.Expression of hepatic proteins was analyzed by immunocytochemistry at day 21.Periodic acid Schiff staining was performed to determine the functional ability of treated cells.RESULTS The fusiform-shaped morphology of MSCs in the treatment groups in comparison with the untreated control,eventually progressed towards the polygonal morphology of hepatocytes with the passage of time.The temporal transcriptional profile of preconditioned MSCs displayed significant expression of hepatic genes with increasing time of differentiation.Preconditioned cells showed positive expression of hepatocyte-specific proteins.The results were further corroborated by positive periodic acid Schiff staining,indicating the presence of glycogen in their cytoplasm.Moreover,bi-nucleated cells,which is the typical feature of hepatocytes,were also seen in the preconditioned cells.CONCLUSION Preconditioning with glycyrrhizic acid,18β-glycyrrhetinic acid and their combination,successfully differentiates hUC-MSCs into hepatic-like cells.These MSCs may serve as a better therapeutic option for degenerative liver diseases in future.

Umbilical cord-derived mesenchymal stem cells preconditioned with isorhamnetin:potential therapy for burn wounds

BACKGROUND Impaired wound healing can be associated with different pathological states.Burn wounds are the most common and detrimental injuries and remain a major health issue worldwide.Mesenchymal stem cells(MSCs)possess the ability to regenerate tissues by secreting factors involved in promoting cell migration,proliferation and differentiation,while suppressing immune reactions.Preconditioning of MSCs with small molecules having cytoprotective properties can enhance the potential of these cells for their use in cell-based therapeutics.AIM To enhance the therapeutic potential of MSCs by preconditioning them with isorhamnetin for second degree burn wounds in rats.METHODS Human umbilical cord MSCs(hU-MSCs)were isolated and characterized by surface markers,CD105,vimentin and CD90.For preconditioning,hU-MSCs were treated with isorhamnetin after selection of the optimized concentration(5μmol/L)by cytotoxicity analysis.The migration potential of these MSCs was analyzed by the in vitro scratch assay.The healing potential of normal,and preconditioned hU-MSCs was compared by transplanting these MSCs in a rat model of a second degree burn wound.Normal,and preconditioned MSCs(IH+MSCs)were transplanted after 72 h of burn injury and observed for 2 wk.Histological and gene expression analyses were performed on day 7 and 14 after cell transplantation to determine complete wound healing.RESULTS The scratch assay analysis showed a significant reduction in the scratch area in the case of IH+MSCs compared to the normal untreated MSCs at 24 h,while complete closure of the scratch area was observed at 48 h.Histological analysis showed reduced inflammation,completely remodeled epidermis and dermis without scar formation and regeneration of hair follicles in the group that received IH+MSCs.Gene expression analysis was time dependent and more pronounced in the case of IH+MSCs.Interleukin(IL)-1β,IL-6 and Bcl-2 associated X genes showed significant downregulation,while transforming growth factorβ,vascular endothelial growth factor,Bcl-2 and matrix metallopeptidase 9 showed significant upregulation compared to the burn wound,showing increased angiogenesis and reduced inflammation and apoptosis.CONCLUSION Preconditioning of hU-MSCs with isorhamnetin decreases wound progression by reducing inflammation,and improving tissue architecture and wound healing.The study outcome is expected to lead to an improved cell-based therapeutic approach for burn wounds.

Regulating the fate of stem cells for regenerating the intervertebral disc degeneration

Lower back pain is a leading cause of disability and is one of the reasons for the substantial socioeconomic burden.The etiology of intervertebral disc(IVD)degeneration is complicated,and its mechanism is still not completely understood.Factors such as aging,systemic inflammation,biochemical mediators,toxic environmental factors,physical injuries,and genetic factors are involved in the progression of its pathophysiology.Currently,no therapy for restoring degenerated IVD is available except pain management,reduced physical activities,and surgical intervention.Therefore,it is imperative to establish regenerative medicine-based approaches to heal and repair the injured disc,repopulate the cell types to retain water content,synthesize extracellular matrix,and strengthen the disc to restore normal spine flexion.Cellular therapy has gained attention for IVD management as an alternative therapeutic option.In this review,we present an overview of the anatomical and molecular structure and the surrounding pathophysiology of the IVD.Modern therapeutic approaches,including proteins and growth factors,cellular and gene therapy,and cell fate regulators are reviewed.Similarly,small molecules that modulate the fate of stem cells for their differentiation into chondrocytes and notochordal cell types are highlighted.

Combination of mesenchymal stem cells and three-dimensional collagen scaffold preserves ventricular remodeling in rat myocardial infarction model

BACKGROUND Cardiovascular diseases are the major cause of mortality worldwide.Regeneration of the damaged myocardium remains a challenge due to mechanical constraints and limited healing ability of the adult heart tissue.Cardiac tissue engineering using biomaterial scaffolds combined with stem cells and bioactive molecules could be a highly promising approach for cardiac repair.Use of biomaterials can provide suitable microenvironment to the cells and can solve cell engraftment problems associated with cell transplantation alone.Mesenchymal stem cells(MSCs)are potential candidates in cardiac tissue engineering because of their multilineage differentiation potential and ease of isolation.Use of DNA methyl transferase inhibitor,such as zebularine,in combination with three-dimensional(3D)scaffold can promote efficient MSC differentiation into cardiac lineage,as epigenetic modifications play a fundamental role in determining cell fate and lineage specific gene expression.AIM To investigate the role of collagen scaffold and zebularine in the differentiation of rat bone marrow(BM)-MSCs and their subsequent in vivo effects.METHODS MSCs were isolated from rat BM and characterized morphologically,immunophenotypically and by multilineage differentiation potential.MSCs were seeded in collagen scaffold and treated with 3μmol/L zebularine in three different ways.Cytotoxicity analysis was done and cardiac differentiation was analyzed at the gene and protein levels.Treated and untreated MSC-seeded scaffolds were transplanted in the rat myocardial infarction(MI)model and cardiac function was assessed by echocardiography.Cell tracking was performed by DiI dye labeling,while regeneration and neovascularization were evaluated by histological and immunohistochemical analysis,respectively.RESULTS MSCs were successfully isolated and seeded in collagen scaffold.Cytotoxicity analysis revealed that zebularine was not cytotoxic in any of the treatment groups.Cardiac differentiation analysis showed more pronounced results in the type 3 treatment group which was subsequently chosen for the transplantation in the in vivo MI model.Significant improvement in cardiac function was observed in the zebularine treated MSC-seeded scaffold group as compared to the MI control.Histological analysis also showed reduction in fibrotic scar,improvement in left ventricular wall thickness and preservation of ventricular remodeling in the zebularine treated MSC-seeded scaffold group.Immunohistochemical analysis revealed significant expression of cardiac proteins in DiI labeled transplanted cells and a significant increase in the number of blood vessels in the zebularine treated MSC-seeded collagen scaffold transplanted group.CONCLUSION Combination of 3D collagen scaffold and zebularine treatment enhances cardiac differentiation potential of MSCs,improves cell engraftment at the infarcted region,reduces infarct size and improves cardiac function.

In vitro modulation of oxidative burst via release of reactive oxygen species from immune cells by extracts of selected tropical medicinal herbs and food plants

Objective:To evaluate in vitro immunomodulating properties and potential cytotoxicity of six tropical medicinal herbs and food plants namely Antidesma madagascariense(Euphorbiaceae) (AM),Erylhroxylum macrocarpum(Erythroxylaceae)(EM),Faujasiopsis flexuosa(Asteraceae) (FF),Pittosporum senacia(Pittosporaceae)(PS),Momordica charantia(Cucurbitaceae)(MC) and Orimum tenuiflorum(Lamiaceae)(Of).Methods:Initially,the crude water and methanol extracts were probed for their capacity to trigger immune cells’ NADPH oxidase and MPO-dependent activities as measured by lucigenin- and luminol-amplified chemiluminescence,respectively; as compared to receptor-dependent(serum opsonised zymosan- OPZ) or receptor-independent phorbol myristerate acetate(PMA).Results:Preliminary screening on whole human blood oxidative buret activity showed significant and concentration-dependent immunomodulating properties of three plants AM,FF and OT.Further investigations of the fractions on isolated human polymorphonuclear cells(PMNs) and mice monocytes using two different pathways for activation of phagocytic oxidative burst showed that ethyl acetate fraction was the most potent extract. None of the active samples had cell-death effects on human PMNs,under the assay conditions as determined by the trypan-blue exclusion assay.Since PMA and OPZ NADPH oxidase complex is activated via different transduction pathways,these results suggest that AM,FF and OT does not affect a specific transductional pathway,but rather directly inhibit a final common biochemical target such as the NADPH oxidase enzyme and/or scavenges ROS.Conclusions:Our findings suggest that some of these plants extracts/fractions were able to modulate significantly immune response of phagocytes and monocytes at different steps,emphasizing their potential as a source of new natural alternative immunomodulatory agents.

The Role of Trace Metals and Environmental Factors in the Onset and Progression of Schizophrenia in Pakistani Population

Exposure to toxic chemicals appears to be one of the major factors in the onset of Schizophrenia. Present study was designed to find out the association of socio-economic, clinical and heavy metals such as chromium (Cr), lead (Pb) copper (Cu), zinc (Zn), iron (Fe), manganese (Mn) selenium (Se) and arsenic (As) as the principle environmental factors that appear to impact schizophrenic condition in Pakistani population. In this study we have tried to expose some factors such as drug abuse, marital status, education, monthly income that could be related to the disease. These parameters were not investigated before in Pakistani subjects. A self-made questionnaire was developed to collect and record the history of patient’s social and economical status. The level of transition metals in the whole blood was also measured by using Inductive couple plasma optical emission spectroscopy (ICP-OES). Present study has found higher levels of Cr, Pb, Zn, Se, As and Cu and lower levels of Fe and Mn in the blood of newly diagnosed (ND) patients when compared to the controls. While comparing ND with the patients who were on medication (Old Diagnosed) we, found lower level of these metals except for copper in ND. However no significant differences were observed between any trace metal levels between the studied groups. The survey concludes that economic status, marital status and illegal drugs are significantly associated with schizophrenia. Pakistani men who use cannabis are significantly higher in numbers when compared to women in acquiring the psychiatric symptoms.

模拟失重不同时长对大鼠抑郁样行为的影响

目的本研究旨在观察不同持续时间的模拟失重对大鼠抑郁样行为和海马超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的影响,以探究其影响及可能的作用机制。方法采用后肢悬挂(HLS)尾吊法模拟大鼠失重状态。将大鼠分为对照组和不同模拟失重时间尾吊组(尾吊时长分别为1、2、3、4周)。采用旷场实验(OFT)、新物体识别实验(NORT)、强迫游泳实验(FST)观察大鼠抑郁样行为,采用酶联免疫吸附试验(ELISA)法测定海马SOD和CAT活性。结果OFT结果显示,与对照组相比,HLS不同时间大鼠的僵滞时间增加(P<0.05,P<0.01)。在NORT中,与对照组相比,HLS不同时间大鼠对新物体的探索潜伏期增加,探索次数和时间减少(P<0.05,P<0.01)。在FST中,与对照组相比,HLS不同时间大鼠在FST中的不动时间增加,攀爬次数减少(P<0.05,P<0.01)。与对照组相比,HLS不同时间组大鼠海马组织中SOD和CAT水平均下降(P<0.05,P<0.01)。结论短时间或长时间的失重都会导致大鼠产生类抑郁样行为。

Evaluation of the risk factors for severe complications and surgery of intestinal foreign bodies in adults:a single-center experience with 180 cases

Background Foreign bodies(FBs)lodged in the intestine or causing intestinal complications are uncommon in clinical practice but may pose diagnostic difficulties and prove life-threatening.This study aimed to evaluate the risk factors for severe complications and surgery to aid clinicians in the diagnosis and management of intestinal FBs.Methods We performed a retrospective analysis of patients in whom FBs were lodged in the intestine or caused complications from 2010 to 2020 in the First Affiliated Hospital of Wenzhou Medical University(Zhejiang,China).The characteristics of the patients and FBs,symptoms,imaging findings,diagnostics,treatment strategies,and clinical outcomes were analysed.Furthermore,the risk factors for complications and surgery were investigated.Results In total,180 patients were included in our study.Most patients(76.1%)were unable to provide a history of ingestion.Bezoars were the most common FBs(35.6%).The FBs were mainly located in the duodenum(32.8%)and the ileum(27.8%).Surgical removal of FBs was successful in 89(49.4%)patients and endoscopic removal in 54(30.0%)patients.Eleven with perforations were treated conservatively.FBs located in the jejunum or ileum were more likely to cause severe complications than those located in the duodenum.FBs located in the jejunum,ileum,or sigmoid colon were more likely to undergo surgery,and severe complications were an independent risk factor for surgery.Conclusion Intestinal FBs,often localized in angulation,are likely to be misdiagnosed because most patients do not provide a history of FB ingestion.Surgery and endoscopic therapy are the most commonly used treatment modalities.Surgery is not mandatory in clinically stable patients with small and contained perforations.FBs located in the jejunum or ileum are risk factors for both complications and surgery.

糖尿病所致认知障碍与中药防治概况

2型糖尿病(T2DM)通常影响老年人群,也是世界上最普遍的糖尿病类型.临床研究表明,T2DM与认知损害,尤其是学习记忆能力下降有关.与正常人群相比,T2DM患者继发痴呆症的风险较高.本文综述了近年来有关T2DM患者认知损害和动物模型方面的研究,包括相关机制和中药治疗的可能性,以期对T2DM并发认知损害有更深入的认识.

槟榔子中抗血小板聚集及抑制乙酰胆碱酯酶活性的有效成分(英文)

目的:研究槟榔子(Areca catechu)粗提取物中所含的抗血小板聚集及抑制乙酰胆碱酯酶活性的有效成分及其作用机制。方法:使用70%甲醇水溶液对槟榔子进行粗提取。使用生物发光血小板凝集仪在富血小板血浆中测定槟榔子粗提取物的抗血小板聚集作用,使用分光光度计在试管内测定槟榔子粗提取物对乙酰胆碱酯酶活性的抑制作用。检测槟榔子中的多种化合物以测定槟榔子中抗血小板聚集及抑制乙酰胆碱酯酶活性的有效成分。结果:槟榔子粗提取物能够抑制花生四烯酸、二磷酸腺苷、血小板活化因子、肾上腺素及钙离子载体引起的血小板聚集,尤其对二磷酸腺苷及钙离子载体引起的血小板聚集的抑制最为明显;槟榔子粗提取物能够显著抑制乙酰胆碱酯酶的活性。在所检测的槟榔子所含化合物中,只有儿茶素对肾上腺素引起的血小板聚集有显著的抑制作用,而这种抑制作用显著弱于槟榔子粗提取物,提示槟榔子中的其他成分参与了这种抑制作用;鞣酸、没食子酸、薯蓣皂苷元和异去甲槟榔次碱能够抑制乙酰胆碱酯酶的活性,其中鞣酸的抑制作用强于槟榔子粗提取物。结论:槟榔子中含有抗血小板聚集及抑制乙酰胆碱酯酶活性的有效成分,而发挥这些功效的确切成分有待进一步的研究证实。

NO-cGMP-K channel-dependent anti-nociceptive activities of methanol stem bark extract of Piptadeniastrum africanum(Mimosaceae) on rats

Objective: To explore anti-hyperalgesic properties of methanol extract of Piptadeniastrum africanum stem bark(PAME) and it possible action mechanism. Methods: PAME was tested on carrageenan ?induced hyperalgesia using plantar test(thermal) and analgesymeter(mechanical) in rats, on prostaglandin E_2(PGE_2) induced mechanical hyperalgesia and vincristine induced neuropathic pain in rat, both with analgesymeter. Modulators of NO/cG MP/K^+ channel pathway and endogenous opioids receptor antagonists and/or agonists were used to determine the possible action mechanism of PAME. Results: PAME significantly decreased carrageenan induced thermal and mechanical hyperalgesia, as well as PGE_2 induced mechanical hyperalgesia. PAME significantly protected the animals against the installation of neuropathic pain. Anti-nociception activity produced by PAME was significantly blocked in animals pre?treated with all the antagonists(naloxone, NW-nitro-L-arginine methyl ester(L-NAME), methylene blue and glibenclamide). Conclusions: Results of this study reveal that, PAME administrate orally, can induce anti-hyperalgesic action against installation of inflammatory pain as well as neuropathic pain. The mechanism underlying PAME antihyperalgesic effect could probably be associated with an activation of opioid receptors and NO/cG MP/K^+ channel pathway.

Evaluation of animal biosafety webinar series for professionals working in animal facilities across Pakistan during the COVID-19 pandemic

Experimental research with animals can help the prevention,cure,and alleviation of human ailments.Animal research facilities are critical for scientific advancement,but they can also pose a higher risk than other biomedical laboratories.Zoonosis,allergic reactions,bites,cuts,and scratches by animals are all substantial concerns that can occur in animal facilities.Furthermore,human error and unexpected animal behavior pose a risk not just to humans,but also to the environment and the animals themselves.The majority of biosafety and biosecurity training programs focus on clinical and biomedical laboratories dealing with human safety factors,with little emphasis on animal biosafety.The current virtual training was designed to improve biosafety and biosecurity capabilities of animal laboratory personnel,researchers,and veterinarians from different regions of Pakistan.The results revealed that understanding was improved regarding triggers for risk assessment in addition to annual and regular reviews(56%to 69%),biosecurity(21%to 50%),decontamination(17%to 35%),safe handling of sharps(21%to 35%),Dual Use Research of Concern(DURC)(17%to 40%),Personal Protective Equipment(PPE)usage by waste handlers(60.9%to 75%),waste management(56%to 85%),animal biosafety levels(40.57%to 45%),and good microbiological practices and procedures(17%to 35%).To bring human and animal laboratories up to the same level in terms of biosafety and biosecurity,it is critical to focus on areas that have been overlooked in the past.Training programs focusing on animal biosafety should be conducted more frequently to strengthen bio risk management systems in animal research facilities.

Ginkgo biloba extract-761 improves the motor functions in permanent middle cerebral artery occlusion rats

OBJECTIVE To study the role of Ginkgo biloba extract-761(EGb-761)in the recovery of gait abnormality and its neuroprotective effect against the brain injury induced by permanent middle cerebral artery occlu-sion in rats.METHODS Male Sprague Dawley rats(n=200,240-305 g)were anesthetized with 0.2%pentobarbital sodium diluted in physiological saline(2.0 m L·kg-1,ip).Then a monofilament coated with poly-L-lysine,was used to occlude the origin of the middle cerebral artery.It was inserted into the internal carotid artery lumen until it met mild resistance,approximately 20mm beyond the common carotid artery bifurcation.The suture was secured with a ligature and maintained in place until sacrifice.The same surgical procedure was conducted in sham-operated rats in which the middle cerebral artery was not occluded.Motor and behavioral changes were assessed after surgery using a five point scale.The rats securing the point scale above 2 were included in the study.The rats were randomly divided into control,and treated groups:EGb-761(20,50,and 100 mg·kg-1).The treated groups were oral y administered(10 mL·kg-1)for 28 d.On 7th,14th,21st,and 28th day the neurological scores,rotar rod test and gait assessment(the automated computer-assisted method)were performed.The brains were collected for TTC staining and histopathological analysis.RESULTS 1)Weight:On 28th day,EGb-761(20 mg·kg-1,)significantly increased the weight of the rat by^8%as compared to control(~300 g).However,at 50 mg·kg-1,and 100 mg·kg-1,a significant increase of^7-7.6%(control:~232 g),and^7.3-7%,respectively from 14 to 28 days was noted.2)Neurological scores:On 28thday,EGb-761(20,50,and 100 mg·kg-1)significantly decreased the neurological scores by^18%,~22%,~21%,respectively as compared to control(~2.07).3)Rotar rod test:On 28thday,EGb-761(50,and100 mg·kg-1)significantly increased by^69.1%,~74.1%,respectively as compared to control(~28.2).4)Gait assessment:On 7th,14th,21st,and 28thday,EGb-761(20,50,and 100 m·kg-1)significantly reduced the average body angle,on 7th,14th,21st,and 28thday,EGb-761(100 mg·kg-1)significantly increased the walk speed and reduced the average walking cycle,EGb-761(50,and 100 mg·kg-1)significantly the area of the left brain/right brain area percentage and reduced tissue pathologic neuron injury.CONCLUSION Ginkgo biloba extract EGb-761 has obvious improve behavior disorders,and has a protective neuroprotective effect against the brain injury induced by permanent middle cerebral artery occlusion.