Development of cryptotanshinone-loaded pellets for angina chronotherapy:In vitro/in vivo prediction and evaluation

The clinical manifestations of variant angina is unevenly distributed during the 24 h, thusthe in vivo performance of drugs should be tailored according to the angina circadianrhythm. Cryptotanshinone(CTN) is one of the representative bioactive lipid-soluble com-ponents of Danshen which has been commonly used for cardiovascular diseases such asangina pectoris. The aim of this study was to develop a novel CTN sustained-released pel-lets(CTN-SRPs) to precisely synchronize the CTN plasma concentrations with predictedoccurrence of angina pectoris for angina chronotherapy. A deconvolution-based methodwas applied to develop and optimize the CTN-SRPs. The plasma concentration-time curveof CTN immediate-released formulation after oral administration in rats was used as theweight function. The predicted plasma concentration-time curve of CTN-SRPs simulatedaccording to the incidence of variant angina during 24 h was used as the response func-tion. Then the desired drug release profile of CTN-SRPs was calculated based on deconvo-lution using weight function and response function, and subsequently used for guiding theformulation optimization. CTN-SRPs were prepared with the combinations of PVP, polox-amer 127 and EC as matrix using fluidized bed technology. An orthogonal design was em-ployed to obtain the optimal formulation with its release profile similar with the desiredone. Pharmacokinetic studies validated that the actual plasma concentration-time curve ofthese optimized CTN-SRPs was similar with the predicted one. In addition, the percent er-rors(%PE) of CTN plasma concentrations in 8–12 h were less than 10%. In conclusion, thisdeconvolution-based method could be applied to adjust the in vivo performance of drugs forangina chronotherapy.

Effects of Chronotherapy of Benazepril on the Diurnal Profile of RAAS and Clock Genes in the Kidney of 5/6 Nephrectomy Rats

Summary： This study investigated the effects of benazepril administered in the morning or evening on the diurnal variation of renin-angiotensin-aldosterone system （RAAS） and clock genes in the kidney. The male Wistar rat models of 5/6 subtotal nephrectomy （STNx） were established. Animals were ran- domly divided into 4 groups： sham STNx group （control）, STNx group, morning benazepril group （MB） and evening benazepril group （EB）. Benazepril was intragastfically administered at a dose of 10 mg/kg/day at 07：00 and 19：00 in the MB group and EB group respectively for 12 weeks. All the animals were synchronized to the light：dark cycle of 12：12 for 12 weeks. Systolic blood pressure （SBP）, 24-h urinary protein excretion and renal function were measured at 11 weeks. Blood samples and kidneys were collected every 4 h throughout a day to detect the expression pattern of renin activity （RA）, angio- tensin Ⅱ （Ang Ⅱ ） and aldosterone （Aid） by radioimmunoassay （RIA） and the mRNA expression profile of clock genes （bmall, dbp and per2） by real-time PCR at 12 weeks. Our results showed that no signifi- cant differences were noted in the SBP, 24-h urine protein excretion and renal function between the MB and EB groups. There were no significant differences in average Aid and RA content of a day between the MB group and EB group. The expression peak of bmall mRNA was phase-delayed by 4 to 8 h, and the diurnal variation of per2 and dbp mRNA diminished in the MB and EB groups compared with the control and STNx groups. It was concluded when the similar SBP reduction, RAAS inhibition and clock gene profile were achieved with optimal dose of benazepril, morning versus evening dosing of benazepril has the same renoprotection effects.

A New Approach: Chronotherapy in Acute Blood Purification for Septic Shock

Circadian rhythms are daily oscillations of multiple biological processes. Recently, relationships between circadian rhythms and immune functions have also been described. In a mouse sepsis model, the death rate due to lipopolysaccharide (LPS)-induced endotoxic shock was found to be dependent on LPS administration as determined by circadian time. In humans, a pronounced inflammatory response to endotoxemia differs depending on whether it is daytime or night-time: Levels of tumor necrosis factor-alpha and interleukin-6 were higher during the night. Therefore, it is reasonable to assume that circadian rhythms influence not only organ dysfunction and the prognosis induced by LPS, but also the therapeutic effect of anti-LPS therapy such as Polymyxin-B direct hemoperfusion. We herein postulate the concept that it is important to discuss septic shock treatment in terms of whether or not the treatment is adjusted for the optimal time window as determined by circadian rhythms.

Circadian rhythm disruption and endocrine-related tumors

This review delved into the intricate relationship between circadian clocks and physiological processes,emphasizing their critical role in maintaining homeo-stasis.Orchestrated by interlocked clock genes,the circadian timekeeping system regulates fundamental processes like the sleep-wake cycle,energy metabolism,immune function,and cell proliferation.The central oscillator in the hypothalamic suprachiasmatic nucleus synchronizes with light-dark cycles,while peripheral tissue clocks are influenced by cues such as feeding times.Circadian disruption,linked to modern lifestyle factors like night shift work,correlates with adverse health outcomes,including metabolic syndrome,cardiovascular diseases,infec-tions,and cancer.We explored the molecular mechanisms of circadian clock genes and their impact on metabolic disorders and cancer pathogenesis.Specific associ-ations between circadian disruption and endocrine tumors,spanning breast,ovarian,testicular,prostate,thyroid,pituitary,and adrenal gland cancers,are highlighted.Shift work is associated with increased breast cancer risk,with PER genes influencing tumor progression and drug resistance.CLOCK gene expression correlates with cisplatin resistance in ovarian cancer,while factors like aging and intermittent fasting affect prostate cancer.Our review underscored the intricate interplay between circadian rhythms and cancer,involving the regulation of the cell cycle,DNA repair,metabolism,immune function,and the tumor microenvir-onment.We advocated for integrating biological timing into clinical consider-ations for personalized healthcare,proposing that understanding these connec-tions could lead to novel therapeutic approaches.Evidence supports circadian rhythm-focused therapies,particularly chronotherapy,for treating endocrine tumors.Our review called for further research to uncover detailed connections between circadian clocks and cancer,providing essential insights for targeted treatments.We emphasized the importance of public health interventions to mitigate lifestyle-related circadian disruptions and underscored the critical role of circadian rhythms in disease mechanisms and therapeutic interventions.

Carcinogenic efects of circadian disruption: an epigenetic viewpoint

Circadian rhythms refer to the endogenous rhythms that are generated to synchronize physiology and behavior with 24-h environmental cues. These rhythms are regulated by both external cues and molecular clock mechanisms in almost all cells. Disruption of circadian rhythms, which is called circadian disruption, afects many biological processes within the body and results in diferent long-term diseases, including cancer. Circadian regulatory pathways result in rhythmic epigenetic modiications and the formation of circadian epigenomes. Aberrant epigenetic modiications, such as hypermethylation, due to circadian disruption may be involved in the transformation of normal cells into cancer cells. Several studies have indicated an epigenetic basis for the carcinogenic efects of circadian disruption. In this review, I irst discuss some of the circadian genes and regulatory proteins. Then, I summarize the current evidence related to the epigenetic modiications that result in circadian disruption. In addition, I explain the carcinogenic efects of circadian disruption and highlight its potential role in diferent human cancers using an epigenetic viewpoint. Finally, the importance of chronotherapy in cancer treatment is highlighted.

The role of circadian rhythm in breast cancer

The circadian rhythm is an endogenous time keeping system shared by most organisms. The circadian clock is comprised of both peripheral oscillators in most organ tissues of the body and a central pacemaker located in the suprachiasmatic nucleus （SCN） of the central nervous system. The circadian rhythm is crucial in maintaining the normal physiology of the organism including, but not limited to, cell proliferation, cell cycle progression, and cellular metabolism; whereas disruption of the circadian rhythm is closely related to multi-tumorigenesis. In the past several years, studies from different fields have revealed that the genetic or functional disruption of the molecular circadian rhythm has been found in various cancers, such as breast, prostate, and ovarian. In this review, we will investigate and present an overview of the current research on the influence of circadian rhythm regulating proteins on breast cancer.

Reversal of Cancer by Dual Strategy?

I present herewith a concise historic account of our development of the new technique for cancer therapy: Reversal of Cancer by Dual Strategy. This new method is based on the possible correction of the mitochondrial filamentation defect of tumors plus minor cytotoxicity addressed to the anoxic and fagocitic tumor’s redoubts, using the administration of the anticancer agents with a chrono-therapeutic emphasis on the patient’s mobility. Such methods probably will reach patients in the near future with the collaboration of private clinical trials mainly at least in the USA, Europe and Japan. Only the more essential references of this effort are provided.

Design and <i>in Vitro</i>Evaluation of Eudragit^®S100/Lipid Based Simvastatin Chronotherapeutic Drug Delivery System

The present study was undertaken to 1) formulate a pulsatile colonic delivery of simvastatin (SIM) as chronotherapy for treatment of hypercholesterolemia, and 2) enhance the dissolution profile of the prepared SIM chronotherapeutic system. Lipid based formulations were utilized to formulate SIM in capsule dosage form coated with Eudragit&reg S100. SIM was formulated using different percentages of Cremophor EL40, Capmul MCM EP and PEG 400. SIM coated capsules (SIMcc) were evaluated for drug release in different pH media. The results showed that SIMcc were able to withstand the acidic pH for 2 hours. Drug release rate was higher (88%) from SIMcc containing 10% polyethylene glycol (PEG) 400. In conclusion, Eudragit&reg S100 as time-dependent and site specific polymer retards SIM release from coated capsules;hence SIMcc could be considered as successful pulsatile treatment of hypercholesterolemia. Also, dissolution profile of lipid based SIMcc was enhanced in comparison with that of SIM filled capsules.

The role of circadian clocks in cancer:Mechanisms and clinical implications

Circadian rhythm refers to the inherent 24-h cycle oscillation of biochemical,phys-iological and behavioral functions,which is almost universal in eukaryotes.At least 14 core clock genes have been reported to form multiple chain feedback loops that confer intrinsic circadian rhythmicity onto the molecular clock.Accumulating evidence has shown that the circadian gene dysfunction resulted from single nucleotide polymorphisms(SNPs),deletions,epigenetic modification,and deregulation is strongly associated with cancer risk.In the pre-sent review,we describe the composition of circadian rhythm system.We highlight the func-tion and mechanism of clock genes in cancer pathogenesis and progression.Moreover,their potential clinical implications as prognostic biomarkers and therapeutic targets have been ad-dressed.

Circadian rhythm in prostate cancer: time to take notice of the clock

The circadian clock is an evolutionary molecular product that is associated with better adaptation to changes in the external environment.Disruption of the circadian rhythm plays a critical role in tumorigenesis of many kinds of cancers,including prostate cancer(PCa).Integrating circadian rhythm into PCa research not only brings a closer understanding of the mechanisms of PCa but also provides new and effective options for the precise treatment of patients with PCa.This review begins with patterns of the circadian clock,highlights the role of the disruption of circadian rhythms in PCa at the epidemiological and molecular levels,and discusses possible new approaches to PCa therapy that target the circadian clock.

Magnetism and Acupuncture

Qi as well as its ability to cure ailments can be released without needling by the application of magnetism. Qi-effects are based upon the magnetic force released by magnetic platelets fastened onto acupoints. This property supports and proves one of the definitions of qi which says, that ＂qi is bio-electrical magnetism＂ or ＂qi is defined as any type of energy which is able to demonstrate power and strength. This energy can be electricity, magnetism, heat, or light＂. Furthermore, it is suggested to use magnet acupuncture parallel to needle acupuncture according to traditional Chinese body acupuncture rules scientifically controlled in order to carry out research on qi and qi-effects. Here it might even be possible to find a quality and even a parameter to measure the intensity of qi-effects, such as by simply measuring the possible change of the heart beat frequency （pulse frequency） before and after application of the needle or magnetic platelet. It is also possible that there is another parameter in existence which will be changed by the impact of qi and acupuncture which still needs to be detected.

Qi,Time and Acupuncture

There is a strong relation among qi, time, and acupuncture. This relation is so close, that it can be said it is of yin-yang nature. Qi stands for yin, time for yang. This fact and its application in acupuncture are known since the times, the Huang Di Nei Jing （Yellow Emperor＇s Inner Canon） was written. The theory of time-related acupuncture, referred to as traditional Chinese chronotherapeutics or chronoacupuncture. In order to achieve better results in acupuncture, time-related acupuncture methods as represented in traditional Chinese chronotherapeufics should be applied in clinic regularly. Furthermore, they should become compulsory for the study of acupuncture.

Flash Acupuncture:A Time-related Acupuncture Method with Magnetic Platelets

Objective：To introduce an acupuncture method called flash acupuncture which combines Ashi points acupuncture with the time-related acupuncture based on traditional Chinese Medicine theories.Methods：Flash acupuncture is carried out with magnetic platelets （6 000 Gauss） mainly at Ashi points,preferably with double magnetic platelets （DMP）,which means that 2 magnetic platelets are put together one above the other.Results：In the case of sciatica,satisfactory curative result was achieved within seconds andminutes with this integrated acupuncture method.Conclusion：For the efficient action and convenience of flash acupuncture,it is worth popularization and application in clinic for treating various diseases.

Shu-Stream Points for Two Cases with Time-related Disease

To introduce two cases with orthopedic and traumatic diseases attacking at specific time periods treated by the Shu-Stream points,and explain the theory‘select Shu-Stream points for condition that alleviate or aggravate in certain time periods’that was recorded in Ling Shu(Spiritual Pivot).

The interaction between circadian rhythm and epilepsy

Evidence about the interaction between circadian rhythms(CR)and epilepsy has been expanded with the application of advanced detection technology.An adequate understanding of how circadian system and epilepsy interact with each other could contribute to more accurate seizure prediction as well as rapid development of potential treatment timed to specific phases of CR.In this review,we present the reciprocal relationship between CR and epileptic activities from aspects of sleep effect,genetic modulation and brain biochemistry.It has been found that sleep-wake patterns,circadian timing systems and multidien rhythms have essential roles in seizure activities and interictal epileptiform discharge(IED).For instance,specific distribution patterns of seizures and IED have been reported,i.e.,lighter non-rapid eye movement(NREM)sleep stage(stage 2)induces seizures while deeper NREM sleep stage(stage 3)activates IEDs.Furthermore,the epilepsy type,seizure type and seizure onset zone can significantly affect the rhythms of seizure occurrence.Apart from the common seizure types,several specific epilepsy syndromes also have a close correlation with sleep-wakefulness patterns.Sleep influences the epilepsy rhythm,and conversely,epilepsy alters the sleep rhythm through multiple pathways.Clock genes accompanied by two feedback loops of regulation have an important role in cortical excitability and seizure occurrence,which may be involved in the mTORopathy.The suprachiasmatic nuclei(SCN)has a rhythm of melatonin and cortisol secretion under the circadian pattern,and then these hormones can feed back into a central oscillator to affect the SCN-dependent rhythms,leading to variable but prominent influence on epilepsy.Furthermore,we discuss the precise predictive algorithms and chronotherapy strategies based on different temporal patterns of seizure occurrence for patients with epilepsy,which may offer a valuable indication for non-invasive closed-loop treatment system.Optimization of the time and dose of antiseizure medications,and resynchronization of disturbed CR(by hormone therapy,light exposure,ketogenic diet,novel small molecules)would be beneficial for epileptic patients in the future.Before formal clinical practice,future large-scale studies are urgently needed to assist prediction and treatment of circadian seizure activities and address unsolved restrictions.