Anti-Mycobacterial Activity of Medicinal Plant Extracts Used in the Treatment of Tuberculosis by Traditional Medicine Practitioners in Uganda

Tuberculosis (TB) remains a public health challenge and one of the leading causes of death worldwide. TB is preventable and curable. However, treatment of tuberculosis has continued to be difficult as a result of rapid increase of multidrug and extensively drug resistant strains of Mycobacterium tuberculosis. Medicinal plants have for centuries been traditionally used in treatment of tuberculosis and similar ailments. They possess antimicrobial properties which render them a new hope as a source of novel bioactive leads in the development of antimycobacterial agents. In this study, 2 plant species commonly used traditionally in Uganda for treatment of tuberculosis, Zanthoxylum leprieurii and Rubia cordifolia were screened for in vitro antimycobacterial activity against Mycobacterium tuberculosis strains;pan sensitive MTB H37Rv, Rifampicin resistant TMC 331 strain and two wild strains (one rifampicin resistant and another one rifampicin susceptible). Antimycobacterial activity of aqueous, ethanolic and methanolic plant extracts was determined using Resazurin Microtiter Assay (REMA). Both plant extracts exhibited significant in vitro antimycobacterial activity against all strains of Mycobacterium tuberculosis. Minimum inhibitory concentrations (MIC) of methanolic crude extracts of both plants ranged from 23.4 μg/mL to 187.5 μg/mL. Comparatively, methanol extracts of both plants possessed superior antimycobacterial activity against all Mycobacterium tuberculosis strains. Our findings indicated that both plants exhibited activity against susceptible and resistant strains of Mycobacterium tuberculosis. While antimycobacterial activity of Z. leprieurii confirms results from previous studies, activity of the extracts of R. cordifolia is reported for the first time in East Africa. Further studies aimed at determining the effects of combination of these plant extracts and standard anti-TB drugs should be carried out.

Molecular mechanism of therapeutic approaches for human gastric cancer stem cells

Gastric cancer(GC)is a primary cause of cancer-related mortality worldwide,and even after therapeutic gastrectomy,survival rates remain poor.The presence of gastric cancer stem cells(GCSCs)is thought to be the major reason for resistance to anticancer treatment(chemotherapy or radiotherapy),and for the development of tumor recurrence,epithelial–mesenchymal transition,and metastases.Additionally,GCSCs have the capacity for self-renewal,differentiation,and tumor initiation.They also synthesize antiapoptotic factors,demonstrate higher performance of drug efflux pumps,and display cell plasticity abilities.Moreover,the tumor microenvironment(TME;tumor niche)that surrounds GCSCs contains secreted growth factors and supports angiogenesis and is thus responsible for the maintenance of the growing tumor.However,the genesis of GCSCs is unclear and exploration of the source of GCSCs is essential.In this review,we provide up-todate information about GCSC-surface/intracellular markers and GCSC-mediated pathways and their role in tumor development.This information will support improved diagnosis,novel therapeutic approaches,and better prognosis using GCSC-targeting agents as a potentially effective treatment choice following surgical resection or in combination with chemotherapy and radiotherapy.To date,most anti-GCSC blockers when used alone have been reported as unsatisfactory anticancer agents.However,when used in combination with adjuvant therapy,treatment can improve.By providing insights into the molecular mechanisms of GCSCs associated with tumors in GC,the aim is to optimize anti-GCSCs molecular approaches for GC therapy in combination with chemotherapy,radiotherapy,or other adjuvant treatment.

Anatomy Education: Perceptions and Experiences of Nigerian Anatomists towards Contributing to Programme and Career Advancements

Anatomy, a foremost basic medical science (BMS) has evolved in content and context, while remaining the most fundamental BMS subject. Stakeholders have continually made efforts especially in recent time to achieve an evolution of this subject. The primary aim of this work was to study the perceptions and experiences of Anatomists towards contributing to programme and career advancements. Following ethical approval, structured questionnaire was administered to Nigerian anatomists using a Google form. It is a total population study. The questionnaire had 13 sections, with each section addressing a theme that was integral to the subject of research interest. The last section collected free responses as qualitative information. After indicating informed consent, 106 anatomists properly completed and returned the questionnaires. Statistical and quantitative analyses of results were done. The most popular field amongst respondents was Neuroscience (60.9%);followed by Histology or Microscopic Anatomy (53.6%), Embryology or developmental anatomy (35.5%), Gross anatomy (34.5%) and Histochemistry (30.9%). About half of Nigerian anatomists involved in the study had a PhD degree. Training emphasized the cognitive domains the most. Consequently, emphasis should be laid on the psychomotor (skills) and the affective domain (attitude) to improve capacity, competences, and the job prospects of trainees. The consensus was that the programme trained students and prepared graduates as medical educators, scientists, and researchers. Anatomy has significant potential to contribute to development with proper programme design, emphasis on research, teaching and applied services competencies, effective policy formulation and implementation and adherence to best practices.

Tumor progression-dependent angiogenesis in gastric cancer and its potential application

Despite improvements in the early diagnosis,prognosis and therapeutic strategies for gastric cancer(GC),human GC remains one of the most frequently diagnosed malignant tumors in the world,and the survival rate of GC patients remains very poor.Thus,a suitable therapeutic strategy for GC is important for prolonging survival.Both tumor cells themselves and the tumor microenvironment play an important role in tumorigenesis,including angiogenesis,inflammation,immunosuppression and metastasis.Importantly,these cells contribute to gastric carcinogenesis by altering the angiogenic phenotype switch.The development,relapse and spreading of tumors depend on new vessels that provide the nutrition,growth factors and oxygen required for continuous tumor growth.Therefore,a state of tumor dormancy could be induced by blocking tumor-associated angiogenesis.Recently,several antiangiogenic agents have been identified,and their potential for the clinical management of GC has been tested.Here,we provide an up-to-date summary of angiogenesis and the angiogenic factors associated with tumor progression in GC.We also review antiangiogenic agents with a focus on the anti-vascular endothelial growth factor receptor(VEGFR)-mediated pathway for endothelial cell growth and their angiogenesis ability in GC.However,most antiangiogenic agents have reported no benefit to overall survival(OS)compared to chemotherapy alone in local or advanced GC.In phase III clinical trials,only ramucirumab(anti-VEGFR blocker)and apatinib(VEGFR-TKI blocker)have reported an improved median overall response rate and prolonged OS and progression-free survival outcomes as a 2 nd-line agent combined with chemotherapy treatment in advanced GC.By providing insights into the molecular mechanisms of angiogenesis associated with tumor progression in GC,this review will hopefully aid the optimization of antiangiogenesis strategies for GC therapy in combination with chemotherapy and adjuvant treatment.

Changes in neuronal apoptosis and apoptosis modulatory factors in cerebral ischemia/reperfusion

BACKGROUND： The high concentration of glutamate release is the main cause for neuronal cell death. The relationship between glutamate level and apoptosis during ischemia/reperfusion injury is still unclear. OBJECTIVE： To observe the neuronal apoptosis at 24 and 72 hours following cerebral ischemia/reperfusion in rats, and analyze the possible influencing factors. DESIGN： A randomized controlled animal experiment. SETTING： School of Medicine, Southern Yangtze University. MATERIALS： Totally 30 male adult Sprague Dawley （SD） rats of clean grade, weighing 240 - 290 g, were obtained from Shanghai Experimental Animal Center, Chinese Academy of Sciences. The rats were randomly divided into sham-operated group （n=10） and model group （n=20）. Each group was observed at 24 and 72 hours after ischemia/reperfusion, 5 rats at each time point in the sham-operated group, whereas 12 at 24 hours and 8 at 72 hours in the model group. Kits for determining apoptosis and Bcl-2 were bought from Wuhan Boster Biological Technology, Co., Ltd.; Kit for calcineurin from Nanjing Jiancheng Bioengineering Institute. METHODS： The experiment was carried out in the Functional Scientific Research Room of Southern Yangtze University from June to October in 2006.①Right middle cerebral artery was occluded by inserting a thread through internal carotid artery （ICA）. The surgical process for the sham-operated rats was the same as that in the model group except a nylon suture inserted the ICA. According to Longa five-degree standard, the neurological deficit evaluation of rats was evaluated after surgery, and grades 1 - 3 were taken as successful model establishment. The blood was recirculated by withdrawing the nylon filament under anesthesia at 2 hours after ischemia in successful rat models. ②After reperfusion, the brain tissue was quickly removed at 24 or 72 hours and the slices were obtained from optic chiasma to funnel manubrium. The changes of the number of apoptotic cells were observed using the terminal deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling method. The expressions of Bcl-2 protein were determined with immunohistochemical staining. The activity of calcineurin was determined by the inorganic phosphorus method. The content of excitatory amino acid was detected by high performance liquid chromatography. MAIN OUTCOME MEASURES： ①Glutanate content in brain tissue; ② Conditions of apoptosis; ③Calcineurin activity in brain tissue; ④ Bcl-2 expression in brain tissue. RESULTS： Totally 30 SD rats were used, 5 died and the other 25 were involved in the analysis of results. ① Changes of apoptosis： There were 0 - 3 apoptotic cells in the sham-operated group. In the model group, the numbers of apoptotic cells were obviously increased at 24 and 72 hours of reperfusion （P 〈 0.01 ）, and it was markedly reduced at 72 hours as compared with 24 hours （P 〈 0.0l）. ② Changes of glutanate content： The glutamate contents at 24 and 72 hours of repeffusion in the model group were obviously higher than those in the sham-operated group （P 〈 0.01）; In the model group, it was obviously increased at 24 hours as compared with 72 hours （P 〈0.01）. ③ Changes of Bcl-2 protein： In the model group, the Bcl-2 protein expression had no obvious changes at 24 hours of reperfusion, and it was obviously enhanced at 72 hours, which was obviously different from that in the sham-operated group and that at 24 hours （P 〈 0.0l）. ④Changes of calcinerin activity： In the model group, the activity of calcineurin in brain tissue had no obvious changes at 24 hours of reperfusion; The activity of calcineurin at 72 hours was obviously higher than that in the sham-operated group and that at 24 hours （P 〈 0.01）. CONCLUSION： The brain cyto-apoptosis action at different time points following reperfusion incompletely depends on the glutamate levels, while it depends on the interaction of some apoptosis related factors, such as amino acid, calcineurin, and Bcl-2, etc.

Quercetin exerts anti-inflammatory effects via inhibiting tumor necrosis factor-α-induced matrix metalloproteinase-9 expression in normal human gastric epithelial cells

BACKGROUND Gastric injury is the most common digestive system disease worldwide and involves inflammation,which can lead to gastric ulcer or gastric cancer(GC).Matrix metallopeptidase-9[MMP-9(gelatinase-B)]plays an important role in inflammation and GC progression.Quercetin and quercetin-rich diets represent potential food supplements and a source of medications for treating gastric injury given their anti-inflammatory activities.However,the effects and mechanisms of action of quercetin on human chronic gastritis and whether quercetin can relieve symptoms remain unclear.AIM To assess whether tumor necrosis factor-α(TNF-α)-induced MMP-9 expression mediates the anti-inflammatory effects of quercetin in normal human gastric mucosal epithelial cells.METHODS The normal human gastric mucosa epithelial cell line GES-1 was used to establish a normal human gastric epithelial cell model of TNF-α-induced MMP-9 protein overexpression to evaluate the antiinflammatory effects of quercetin.The cell counting Kit-8 assay was used to evaluate the effects of varying quercetin doses on cell viability in the normal GES-1 cell line.Cell migration was measured using Transwell assay.The expression of proto-oncogene tyrosine-protein kinase Src(cSrc),phospho(p)-c-Src,extracellular-signal-regulated kinase 2(ERK2),p-ERK1/2,c-Fos,p-c-Fos,nuclear factor kappa B(NF-κB/p65),and p-p65 and the effects of their inhibitors were examined using Western blot analysis and measurement of luciferase activity.p65 expression was detected by immunofluorescence.MMP-9 m RNA and protein levels were measured by quantitative reverse transcription polymerase chain reaction(q RT–PCR)and gelatin zymography,respectively.RESULTS q RT-PCR and gelatin zymography showed that TNF-αinduced MMP-9 m RNA and protein expression in a dose-and time-dependent manner.These effects were reduced by the pretreatment of GES-1 cells with quercetin or a TNF-αantagonist(TNFR inhibitor)in a dose-and timedependent manner.Quercetin and TNF-αantagonists decreased the TNF-α-induced phosphorylation of c-Src,ERK1/2,c-Fos,and p65 in a dose-and time-dependent manner.Quercetin,TNF-αantagonist,PP1,U0126,and tanshinone IIA(TSIIA)reduced TNF-α-induced c-Fos phosphorylation and AP-1–Luciferase(Luc)activity in a dose-and time-dependent manner.Pretreatment with quercetin,TNF-αantagonist,PP1,U0126,or Bay 11-7082 reduced TNF-α-induced p65 phosphorylation and translocation and p65–Luc activity in a dose-and timedependent manner.TNF-αsignificantly increased GES-1 cell migration,and these results were reduced by pretreatment with quercetin or a TNF-αantagonist.CONCLUSION Quercetin significantly downregulates TNF-α-induced MMP-9 expression in GES-1 cells via the TNFR-c-Src–ERK1/2 and c-Fos or NF-κB pathways.

Designing and generating a mouse model:frequently asked questions

Genetically engineered mouse(GEM)models are commonly used in biomedical research.Generating GEMs involve complex set of experimental procedures requiring sophisticated equipment and highly skilled technical staff.Because of these reasons,most research institutes set up centralized core facilities where custom GEMs are created for research groups.Researchers,on the other hand,when they begin thinking about generating GEMs for their research,several questions arise in their minds.For example,what type of model(s)would be best useful for my research,how do I design them,what are the latest technologies and tools available for developing my model(s),and finally how to breed GEMs in my research.As there are several considerations and options in mouse designs,and as it is an expensive and time-consuming endeavor,careful planning upfront can ensure the highest chance of success.In this article,we provide brief answers to several frequently asked questions that arise when researchers begin thinking about generating mouse model(s)for their work.

Reduced normogastric electrical activity associated with emesis:A telemetric study in ferrets

AIM:To characterize the gastric myoelectric activity(GMA) and intra-abdominal pressure changes induced by emetic stimuli(apomorphine and cisplatin) in the ferret.METHODS:GMA and intra-abdominal pressure were recorded in conscious,unrestrained ferrets surgically implanted with radiotelemetry transmitters.Animals were challenged with apomorphine(0.25 mg/kg sc) and cisplatin(10 mg/kg ip),and the emetic response was quantif ied via direct observation and intra-abdominal pressure recording for 1 and 4 h,respectively.The GMA was analyzed by spectral analysis;the parameters used to characterize the GMA were the dominant frequency(DF) and the repartition of spectral power in the bradygastric,normogastric and tachygastric frequency ranges.RESULTS:Retches were identified on the intraabdominal pressure trace as peaks 0.30±1.01 s in duration and 59.57±2.74 mmHg in amplitude,vomit peaks were longer(0.82±0.06 s,P<0.01) and reached a higher pressure(87.73±8.12 mmHg,P<0.001).The number of retches and vomits quantified via direct observation [apomorphine:65.5 ± 11.8 retches + vomits(R+V),cisplatin:202.6±64.1 R+V] and intra-abdominal pressure(apomorphine:68.3±13.7 R+V,n=8;cisplatin:219.0±69.2 R+V,n=8) were correlated(r=0.97,P<0.0001) and the timing of emesis was consistent between the 2 methods.Apomorphine induced a decrease in normogastria from 45.48%±4.35% to 36.70±4.34%(n=8,P<0.05) but the DF of the slow waves was not changed [8.95±0.25 counts/min(cpm) vs 8.68±0.35 cpm,n=8,P> 0.05].Cisplatin induced a decrease in normogastria from 55.83%±4.30% to 29.22%±5.16% and an increase in bradygastria from 14.28%±2.32% to 31.19%±8.33%(n=8,P<0.001) but the DF(9.14±0.13 cpm) remained unchanged(P>0.05).The GMA changes induced by cisplatin preceded the emetic response as normogastria was reduced for 1 h before the onset of emesis(57.61%±5.66% to 39.91%±5.74%,n=6,P<0.05).Peri-emesis analysis revealed that the GMA was signif icantly disturbed during and immediately after,but not immediately before,the emetic episodes.CONCLUSION:The induction of emesis is reliably associated with a disrupted GMA,but changes may also occur prior to and following the emetic response.

Specific acupuncture stimulation of Shenshu(BL23)affects sympathetic nervous activity-associated plasma renin concentration changes

OBJECTIVE:To examine whether specific stimulation of Shenshu(BL23)affects sympathetic nervous activity(SNA)-associated plasma renin concentration(PRC).METHODS:Eight healthy volunteers participated in three pattern conditions in random order:control(Cont),stimulation of Shenshu(BL23),and stimulation of sham point(Sham).All participants were initially in the supine position for>60 min,and then remained in the standing position during the experimental procedure to increase SNA.An electrocardiogram was used to calculate low frequency/high frequency(LF/HF)ratio;blood was collected to analyze PRC.RESULTS:The LF/HF ratio was significantly increased in the standing position when compared with the supine position(P<0.01).There was no difference in LF/HF ratio during or after stimulation of Shenshu(BL23)in the standing position when compared with before the stimulation in the supine position;however,the LF/HF ratio was significantly increased in Cont and Sham conditions(P<0.01).There was no difference in PRC after stimulation of Shenshu(BL23)in the standing position when compared with before the stimulation in the supine position;however,there was a significant increase in PRC in the Cont and Sham conditions(Cont P<0.05,Sham P<0.01).CONCLUSION:Our results demonstrated that specific acupuncture stimulation of Shenshu(BL23)in the standing position decreased SNA-associated PRC,which was not observed during acupuncture stimulation of the sham point.