Research on the development methodology for clinical practice guidelines for organic integration of traditional Chinese medicine and Western medicine

Integrated traditional Chinese medicine(TCM)and Western medicine(WM)is a new medical science grounded in the knowledge bases of both TCM and WM,which then forms a unique modern medical system in China.Integrated TCM and WM has a long history in China,and has made important achievements in the process of clinical diagnosis and treatment.However,the methodological defects in currently published clinical practice guidelines(CPGs)limit its development.The organic integration of TCM and WM is a deeper integration of TCM and WM.To realize the progression of"integration"to"organic integration",a targeted and standardized guideline development methodology is needed.Therefore,the purpose of this study is to establish a standardized development procedure for clinical practice guidelines for the organic integration of TCM and WM to promote the systematic integration of TCM and WM research results into clinical practice guidelines in order to achieve optimal results as the whole is greater than the sum of the parts.

Effects of a periodic intermittent theta burst stimulation in Alzheimer's disease

Background Previous studies havedemonstrated that excitatory repetitive transcranial magnetic stimulation(rTMS)can improve the cognitive function of patients with Alzheimer's disease(AD).Intermittent theta burst stimulation(iTBS)is a novel excitatory rTMS protocol for brain activity stimulation with the ability to induce long-term potentiation-like plasticity and represents a promising treatment for AD.However,the long-term effects of iTBS on cognitive decline and brain structure in patients with AD areunknown.Aims We aimed to explore whether repeating accelerated iTBS every three months could slow down the cognitive decline in patients with AD.Methods In this randomised,assessor-blinded,controlled trial,iTBS was administered to the left dorsolateral prefrontal cortex(DLPFC)of 42 patients with AD for 14days every 13weeks.Measurements included the Montreal Cognitive Assessment(MoCA),a comprehensive neuropsychological battery,and the grey matter volume(GMV)of the hippocampus.Patients were evaluated at baseline and after follow-up.The longitudinal pipeline of the Computational Anatomy Toolbox for SPM was used to detect significant treatment-related changes over time.Results The iTBS group maintained MoCA scores relative to the control group(t=3.26,p=0.013)and reduced hippocampal atrophy,which was significantly correlated with global degeneration scale changes.The baseline Mini-Mental State Examination(MMSE)score,apolipoprotein E genotype and Clinical Dementia Rating were indicative of MoCA scores at follow-up.Moreover,the GMV of the left(t=0.08,p=0.996)and right(t=0.19,p=0.977)hippocampus were maintained in the active group but significantly declined in the control group(left:t=4.13,p<0.001;right:t=5.31,p<0.001).GMV change in the left(r=0.35,p=0.023)and right(r=0.36,p=0.021)hippocampus across the intervention positively correlated with MoCA changes;left hippocampal GMV change was negatively correlated with global degeneration scale(r=-0.32,p=0.041)changes.Conclusions DLPFC-iTBS maybe a feasible and easy-to-implement non-pharmacological intervention to slow down the progressive decline of overall cognition and quality of life in patients with AD,providing a new AD treatment option.Trial registration number NCT04754152.

Microglia activation in the offspring of prenatal Poly I:C exposed rats:a PET imaging and immunohistochemistry study

Background The well-known <pyrotherapy, of Julius Wagner-Jauregg might be the beginning of the study on the immunological concepts of schizophrenia. As the primary immune effector cells in the brain, microglia play a pivotal role in neuroinflammatory processes. Maternal viral infection during pregnancy is associated with an increased risk for psychiatric disorders with presumed neurodevelopmental origin, including autism spectrum disorders and schizophrenia. The present study was to quantify microglia activation in vivo in the mature offspring of rats exposed to polyriboinosinic-polyribocytidilicacid(Poly l:C) during pregnancy using 11 C'PK11195 positron emission tomography(PET) and immunohistochemistry.Objective The study aimed to quantify microglia activation in vivo in the prefrontal cortex and hippocampus in mature offspring of prenatal Poly l:C exposed rats.Methods Offspring of Poly l:C-treated dams were the model group, offspring of saline-treated dams were the control group. Behavioural test for two groups was taken by spontaneous activity, prepulse inhibition(PPI) and latent inhibition(LI) test(including active avoidance conditioning task and passive avoidance conditioning task). Randomly selected successful model rats were assessed by behavioural test in the model group and control group rats.11 C-PK11195 micro-PET/CT and immunohistochemistry were performed on the selected rats to measure microglia activation.Results The treatment group showed hyperlocomotion and deficits in PPI and LI compared with the control group. The treatment group also showed an increased11 C-PK11195 uptake ratio in the prefrontal cortex(f=-3.990, p=0.003) and hippocampus(f=-4.462,p=0.001). The number of activated microglia cells was significantly higher in the treatment group than in the control gro叩(hippocampus: f=8.204, p<0.001; prefrontal:f=6.995, p<0.001). Within the treatment group, there were significant correlations between the behavioural parameters and the activation of microglia as measured by PET and immunohistochemistry.Conclusions The present study demonstrated microglia activation in vivo in the prefrontal cortex and hippocampus in mature offspring of prenatal Poly l:C exposed rats.This study suggests that microglia activation may play a possible or potential role in the pathogenesis of schizophrenia.

Variant rs8400 enhances ALKBH5 expression through disrupting miR-186 binding and promotes neuroblastoma progression

Objective:AlkB homolog 5(ALKBH5)has been proven to be closely related to tumors.However,the role and molecular mechanism of ALKBH5 in neuroblastomas have rarely been reported.Methods:The potential functional single-nucleotide polymorphisms(SNPs)in ALKBH5 were identified by National Center for Biotechnology Information(NCBI)dbSNP screening and SNPinfo software.TaqMan probes were used for genotyping.A multiple logistic regression model was used to evaluate the effects of different SNP loci on the risk of neuroblastoma.The expression of ALKBH5 in neuroblastoma was evaluated by Western blotting and immunohistochemistry(IHC).Cell counting kit-8(CCK-8),plate colony formation and 5-ethynyl-2'-deoxyuridine(EdU)incorporation assays were used to evaluate cell proliferation.Wound healing and Transwell assays were used to compare cell migration and invasion.Thermodynamic modelling was performed to predict the ability of miRNAs to bind to ALKBH5 with the rs8400 G/A polymorphism.RNA sequencing,N6-methyladenosine(mA)sequencing,mA methylated RNA immunoprecipitation(MeRIP)and a luciferase assay were used to identify the targeting effect of ALKBH5 on SPP1.Results:ALKBH5 was highly expressed in neuroblastoma.Knocking down ALKBH5 inhibited the proliferation,migration and invasion of cancer cells.miR-186-3p negatively regulates the expression of ALKBH5,and this ability is affected by the rs8400 polymorphism.When the G nucleotide was mutated to A,the ability of miR-186-3p to bind to the 3'-UTR of ALKBH5 decreased,resulting in upregulation of ALKBH5.SPPI is the downstream target gene of the ALKBH5 oncogene.Knocking down SPP1 partially restored the inhibitory effect of ALKBH5 downregulation on neuroblastoma.Downregulation of ALKBH5 can improve the therapeutic efficacy of carboplatin and etoposide in neuroblastoma.Conclusions:We first found that the rs8400 G>A polymorphism in the m6A demethylase-encoding gene ALKBH5 increases neuroblastoma susceptibility and determines the related mechanisms.The aberrant regulation of ALKBH5 by miR-186-3p caused by this genetic variation in ALKBH5 promotes the occurrence and development of neuroblastoma through the ALKBH5-SPP1 axis.

Research Progress on Metabolic Diseases in Rural Areas

Through the review of relevant literature in recent years,it is found that the incidence rate of metabolic diseases in rural areas is on the rise:The related factors that may cause diseases include diet structure,exercise habits,medical resources and other aspects.At the same time,there are some limitations in the intervention and treatment measures of this kind of disease.It is urgent to carry out in-depth exploration for the special conditions in rural areas.By reviewing the research progress related to metabolic diseases in rural areas,more new ideas are provided for the treatment and improvement of metabolic diseases in rural areas.

Exogeneous metal ions as therapeutic agents in cardiovascular disease and their delivery strategies

Metal ions participate in many metabolic processes in the human body,and their homeostasis is crucial for life.In cardiovascular diseases(CVDs),the equilibriums of metal ions are frequently interrupted,which are related to a variety of disturbances of physiological processes leading to abnormal cardiac functions.Exogenous supplement of metal ions has the potential to work as therapeutic strategies for the treatment of CVDs.Compared with other therapeutic drugs,metal ions possess broad availability,good stability and safety and diverse drug delivery strategies.The delivery strategies of metal ions are important to exert their ther-apeutic effects and reduce the potential toxic side effects for cardiovascular applications,which are also receiving increasing attention.Controllable local delivery strategies for metal ions based on various biomaterials are constantly being designed.In this review,we comprehensively summarized the positive roles of metal ions in the treatment of cVDs from three aspects:protecting cells from oxidative stress,inducing angiogenesis,and adjusting the functions of ion channels.In addition,we introduced the transferability of metal ions in vascular reconstruction and cardiac tissue repair,as well as the currently available engineered strategies for the precise delivery of metal ions,such as integrated with nanoparticles,hydrogels and scaffolds.

Structure,ligands,and roles of GPR126/ADGRG6 in the development and diseases

Adhesion G protein-coupled receptors(aGPCRs)are the second largest diverse group within the GPCR superfamily,which play critical roles in many physiological and patho-logical processes through cell-cell and cell-extracellular matrix interactions.The adhesion GPCR Adgrg6,also known as GPR126,is one of the better-characterized aGPCRs.GPR126 was previously found to have critical developmental roles in Schwann cell maturation and its mediated myelination in the peripheral nervous system in both zebrafish and mammals.Current studies have extended our understanding of GPR126-mediated roles during develop-ment and in human diseases.In this review,we highlighted these recent advances in GPR126 in expression profile,molecular structure,ligand-receptor interactions,and associated physiological and pathological functions in development and diseases.

Two antibodies show broad,synergistic neutralization against SARS-CoV-2 variants by inducing conformational change within the RBD

Continual evolution of the severe acute respiratory syndrome coronavirus(SARS-CoV-2)virus has allowed for its gradual evasion of neutralizing antibodies(nAbs)produced in response to natural infection or vaccination.The rapid nature of these changes has incited a need for the development of superior broad nAbs(bnAbs)and/or the rational design of an antibody cocktail that can protect against the mutated virus strain.Here,we report two angiotensin-converting enzyme 2 competing nAbs—8H12 and 3E2—with synergistic neutralization but evaded by some Omicron subvariants.Cryo-electron microscopy reveals the two nAbs synergistic neutralizing virus through a rigorous pairing permitted by rearrangement of the 472-489 loop in the receptor-binding domain to avoid steric clashing.Bispecific antibodies based on these two nAbs tremendously extend the neutralizing breadth and restore neutralization against recent variants including currently dominant XBB.1.5.Together,these findings expand our understanding of the potential strategies for the neutralization of SARS-CoV-2 variants toward the design of broad-acting antibody therapeutics and vaccines.

Factors associated with failure of enhanced recovery after surgery program in patients undergoing pancreaticoduodenectomy

Background: The enhanced recovery after surgery (ERAS) protocol is an evidence-based perioperative care program aimed at reducing surgical stress response and accelerating recovery. However, a small propor- tion of patients fail to bene t from the ERAS program following pancreaticoduodenectomy. This study aimed to identify the risk factors associated with failure of ERAS program in pancreaticoduodenectomy. Methods: Between May 2014 and December 2017, 176 patients were managed with ERAS program fol-lowing pancreaticoduodenectomy. ERAS failure was indicated by prolonged hospital stay, unplanned read- mission or unplanned reoperation. Demographics, postoperative recovery and compliance were compared of those ERAS failure groups to the ERAS success group. Results: ERAS failure occurred in 59 patients, 33 of whom had prolonged hospital stay, 18 were readmitted to hospital within 30 days after discharge, and 8 accepted reoperation. Preoperative American Society of Anesthesiologists (ASA) score of ≥III (OR = 2.736;95% CI: 1.276 6.939;P=0.028) and albumin (ALB) level of <35g/L (OR=3.589;95% CI: 1.403 9.181;P=0.008) were independent risk factors associated with prolonged hospital stay. Elderly patients (>70 years) were on a high risk of unplanned reoperation (62.5% vs. 23.1%, P=0.026). Patients with prolonged hospital stay and unplanned reoperation had delayed intake and increased intolerance of oral foods. Prolonged stay patients got off bed later than ERAS success patients did (65h vs. 46h, P =0.012). Unplanned reoperation patients tended to experience severer pain than ERAS success patients did (3 score vs. 2 score, P =0.035). Conclusions: Patients with high ASA score, low ALB level or age >70 years were at high risk of ERAS failure in pancreaticoduodenectomy. These preoperative demographic and clinical characteristics are important determinants to obtain successful postoperative recovery in ERAS program.

Androgen receptor expression in clinically localized prostate cancer： immunohistochemistry study and literature review

Aim： To evaluate androgen receptor （AR） expression in clinically localized prostate cancer （PCa）. Methods： Specimens were studied from 232 patients who underwent radical prostatectomy for clinically localized prostatic adenocarcinoma without neoadjuvant hormonal therapy or chemotherapy at our institution between November 2001 and June 2005. Immunohistochemical study was performed using an anti-human AR monoclonal antibody AR441. The mean AR density in the hot spots of different histological areas within the same sections were compared and the correlation of malignant epithelial AR density with clinicopathological parameters such as Gleason score, tumor, nodes and metastases （TNM） stage and pre-treatment prostate-specific antigen （PSA） value was assessed. Results： AR immunoreactivity was almost exclusively nuclear and was observed in tumor cells, non-neoplastic glandular epithelial cells and a proportion of peritumoral and interglandular stromal cells. Mean percentage of AR-positive epithelial cells was significantly higher in cancer tissues than that in normal prostate tissues （mean e SD, 90.0% ± 9.3% vs. 85.3% ±9.7%, P 〈 0.001）. The histological score yielded similar results. The percentage ofAR immunoreactive prostatic cancer nuclei and histological score were not correlated with existing parameters such as Gleason score, tumor, nodes and metastases stage and pre-treatment PSA value in this surgically treated cohort. Conclusion： The results of the present study suggest that there may be limited clinical use for determining AR expression （if evaluated in hot spots） in men with localized PCa.

Genomic Characterization of a Streptococcus suis Serotype 2 Isolated from a Human Patient

Streptococcus suis(S. suis) is a Gram-positive zoonotic pathogen. S. suis infection in humans commonly causes meningitis, septicemia, arthritis,and streptococcal toxic shock-like syndrome(STSLS). S. suis has 29 serotypes, of which S. suis serotype 2(SS2) has the highest clinical isolation rate and strongest pathogenicity and causes most S. suis human infections.About 1,000 different sequence types(STs)were identified in S.suis based on multilocus sequence typing(MLST).Among these,STs,ST1,and ST7 belong to serotype 2 and are the major hazards of S.suis.in 1998 and 2005,two cases of SS2 human infection outbreak were reported in Jiangsu and Sichuan,China,causing 14 deaths and 38 deaths,respectively.

Non-canonical STING-PERK pathway dependent epigenetic regulation of vascular endothelial dysfunction via integrating IRF3 and NF-κB in inflammatory response

Inflammation-driven endothelial dysfunction is the major initiating factor in atherosclerosis,while the underlying mechanism remains elusive.Here,we report that the non-canonical stimulator of interferon genes(STING)-PKR-like ER kinase(PERK)pathway was significantly activated in both human and mice atherosclerotic arteries.Typically,STING activation leads to the activation of interferon regulatory factor 3(IRF3)and nuclear factor-kappa B(NF-κB)/p65,thereby facilitating IFN signals and infammation.In contrast,our study reveals the activated non-canonical STING-PERK pathway increases scaffold protein bromodomain protein 4(BRD4)expression,which encourages the formation of super-enhancers on the proximal promoter regions of the proinflammatory cytokines,thereby enabling the transactivation of these cytokines by integrating activated IRF3 and NF-κB via a condensation process.Endothelium-specific STING and BRD4 deficiency significantly decreased the plaque area and inflammation.Mechanistically,this pathway is triggered by leaked mitochondrial DNA(mtDNA)via mitochondrial permeability transition pore(mPTP),formed by voltage-dependent anion channel 1(VDAC1)oligomer interaction with oxidized mtDNA upon cholesterol oxidation stimulation.Especially,compared to macrophages,endothelial STING activation plays a more pronounced role in atherosclerosis.We propose a non-canonical STING-PERK pathway-dependent epigenetic paradigm in atherosclerosis that integrates IRF3,NF-κB and BRD4 in inflammatory responses,which provides emerging therapeutic modalities for vascular endothelial dysfunction.

Targeting the blood-brain barrier to delay aging-accompanied neurological diseases by modulating gut microbiota,circadian rhythms,and their interplays

AbstracTthe blood-brain barrier(BBB)impairment plays a crucial role in the pathological processes of aging-accompanied neurological diseases(AAND).Meanwhile,circadian rhythms disruption and gut microbiota dysbiosis are associated with increased morbidity of neurological diseases in the accelerated aging population.Importantly,circadian rhythms disruption and gut microbiota dysbiosis are also known to induce the generation of toxic metabolites and pro-inflammatory cytokines,resulting in disruption of BBB integrity.Collectively,this provides a new perspective for exploring the relationship among circadian rhythms,gut microbes,and the BBB in aging-accompanied neurological diseases.In this review,we focus on recent advances in the interplay between circadian rhythm disturbances and gut microbiota dysbiosis,and their potential roles in the BBB disruption that occurs in AAND.Based on existing literature,we discuss and propose potential mechanisms underlying BBB damage induced by dysregulated circadian rhythms and gut microbiota,which would serve as the basis for developing potential interventions to protect the BBB in the aging population through targeting the BBB by exploiting its links with gut microbiota and circadian rhythms for treating AAND.

Formulation,characterization and in vivo and in vitro evaluation of aloe-emodin-loaded solid dispersions for dissolution enhancement

OBJECTIVE:To prepare aloe-emodin solid dispersion(AE-SD)and determine the metabolic process of AE and AE-SD in vivo.METHODS:AE-SD was prepared via solvent evaporation or solvent melting using PEG-6000 and PVP-K30 as carriers.Thermogravimetric analysis,X-ray diffraction spectroscopy,differential scanning calorimetry,Fourier transform infrared spectroscopy and scanning electron microscopy were used to identify the physical state of AESD.Optimal prescriptions were screened via the dissolution degree determination method.Using Phoenix software,AE suspension and AE-SD were subjected to a pharmacokinetic comparison study analyzing the alteration of behavior in vivo after AE was prepared as a solid dispersion.Acute toxicity was assessed in mice,and the physiological toxicity was used as the determination criterion for toxicity.RESULTS:AE-SD showed that AE existed in the carrier in an amorphous state.Compared with polyethylene glycol,polyvinylpyrrolidone(PVP)inhibited AE crystallization,causing the drug to transform from a dense crystalline state to an amorphous form and increasing the degree of drug dispersion.Therefore,it was more suitable as a carrier material for AE-SD.The addition of poloxamer(POL)was more beneficial to the stability of solid dispersions and could reduce the amount of PVP.The dissolution test confirmed that the optimal ratio of AE to the composite vector AE-PVP-POL was 1:2:2,and its dissolution effect was also optimal.Based on the pharmacokinetic comparison,the drug absorption was faster and quickly reached the peak of blood drug concentration in AE-SD compared to AE,the Cmax of AE-SD was greater than that of AE,and t1/2 and mean residence time of AE-SD were less than AE.The results showed that the drug metabolism in AE-SD was better,and the residence time was shorter.The toxicology study showed that both AE and AE-SD had no toxicity.CONCLUSION:This paper established that the solubility of the drug could be increased after preparing a solid dispersion,as demonstrated by in vitro dissolution experiments.In vivo pharmacokinetics studies confirmed that AE-SD could improve the bioavailability of AE in vivo,providing a new concept for the research and development of AE preparations.

Encyclopedia of fecal microbiota transplantation: a review of effectiveness in the treatment of 85 diseases

Fecal microbiota transplantation(FMT)has been used as a core therapy for treating dysbiosis-related diseases by remodeling gut microbiota.The methodology and technology for improving FMT are stepping forward,mainly including washed microbiota transplantation(WMT),colonic transendoscopic enteral tubing(TET)for microbiota delivery,and purified Firmicutes spores from fecal matter.To improve the understanding of the clinical applications of FMT,we performed a systematic literature review on FMT published from 2011 to 2021.Here,we provided an overview of the reported clinical benefits of FMT,the methodology of processing FMT,the strategy of using FMT,and the regulations on FMT from a global perspective.A total of 782 studies were included for the final analysis.The present review profiled the effectiveness from all clinical FMT uses in 85 specific diseases as eight categories,including infections,gut diseases,microbiotagut-liver axis,microbiotagut-brain axis,metabolic diseases,oncology,hematological diseases,and other diseases.Although many further controlled trials will be needed,the dramatic increasing reports have shown the promising future of FMT for dysbiosis-related diseases in the gut or beyond the gut.

A novel lytic phage potentially effective for phage therapy against Burkholderia pseudomallei in the tropics

Background:Burkholderia pseudomallei is a tropical pathogen that causes melioidosis.Its intrinsic drug-resistance is a leading cause of treatment failure,and the few available antibiotics require prolonged use to be effective.This study aimed to assess the clinical potential of B.pseudomallei phages isolated from Hainan,China.Methods:Burkholderia pseudomallei strain(HNBPoo1)was used as the isolation host,and phages were recovered from domestic environmental sources,which were submitted to the host range determination,lytic property assays,and stability tests.The best candidate was examined via the transmission electron microscope for classification.With its genome sequenced and analyzed,its protective efficacy against B.pseudomallei infection in A549 cells and Caenorhabditis elegans was evaluated,in which cell viability and survival rates were compared using the one-way ANOVA method and the log-rank test.Results:A phage able to lyse 24/25 clinical isolates was recovered.It was classified in the Podoviridae family and was found to be amenable to propagation.Under the optimal multiplicity of infection(MOl)of o.1,an eclipse period of around 20 min and a high titer(io12 PFU/ml)produced within 1 h were demonstrated.This phage was found stabile at a wide range of temperatures(24,37,40,50,and 60 C)and pH values(3-12).After being designated as vB_BpP_HN01,it was fully sequenced,and the 71,398 bp linear genome,containing 93 open reading frames and a tRNA-Asn,displayed a low sequence similarity with known viruses.Additionally,protective effects of applications of vB_BpP_HNo1(MOl=0.1 and MOl=1)alone or in combination with antibiotics were found to improve viability of infected cells(70.6±6.8%,85.8±5.7%,91.9±1.8%,and 96.8±1.8%,respectively).A significantly reduced mortality(10%)and a decreased pathogen load were demonstrated in infected C.elegans following the addition of this phage.Conclusions:As the frst B.pseudomallei phage was isolated in Hainan,China,phage vB_BpP_HNO1 was characterized by promising lytic property,stability,and effciency of bacterial elimination during the in vitro/vivo experiments.Therefore,we can conclude that it is a potential alternative agent for combating melioidosis.

Integrin β2 and β3:Two plasmatocyte markers deepen our understanding of the development of plasmatocytes in the silkworm Bombyx mori

Insect hemocytes play important biological roles at developmental stages,metamorphosis,and innate immunity.As one of the most abundant cell types,plasmatocytes can participate in various innate immune responses,especially in encapsulation and node formation.Here,2 molecular markers of plasmatocytes,consisting of integrinβ2 andβ3,were identified and used to understand the development of plasmatocytes.Plasmatocytes are widely distributed in the hematopoietic system,including circulating hemolymph and hematopoietic organs(HPOs).HPOs constantly release plasmatocytes with high proliferative activity in vitro;removal of HPOs leads to a dramatic reduction in the circulating plasmatocytes,and the remaining plasmatocytes gradually lose their ability to proliferate in vivo.Our results demonstrated that the release of plasmatocytes from HPOs is regulated by insulin-mediated signals and their downstream pathways,including PI3K/Akt and MAPK/Erk signals.The insulin/PI3K/Akt signaling pathway can significantly irritate the hematopoiesis,and its inhibitor LY294002 could inhibit the hemocytes discharged from HPOs.While the insulin/MAPK/Erk signaling pathway plays a negative regulatory role,inhibiting its activity with U0126 can markedly promote the discharge of plasmatocytes from HPOs.Our results indicate that the circulating plasmatocytes are mainly generated and discharged by HPOs.This process is co-regulated by the PI3K/Akt and MAPK/Erk signals in an antagonistic manner to adjust the dynamic balance of the hemocytes.These findings can enhance our understanding of insect hematopoiesis.

Gut microbisal methionine impacts circadian clock gene expression and reactive oxygen species level in host gastrointestinal tract

Dear Editor,Circadian rhythms are the periodic turmover of biological behaviors and physiological functions of many organisms(Allada and Bass,2021).In mammals,circadian rhythms are maintained by the clock system,involving central clock in suprachiasmatic nucleus and peripheral clock in organs like intestine;and at cellular level a cell-autonomous transcriptional and translational feedback loop involving clock genes like BMAL1(brain and muscle ARNT-Like 1,also known as ARNTL),CLOCK(clock circadian regulator),PER1/2/3(period circadian regulator 1/2/3)and CRY1/2(cryptochrome circadian regulator 1/2)(Allada and Bass,2021).

Principles of extramural hl ealth care for chronic wounds in China

Development of extramural health care for chronic wounds is still in its infancy in China,and thus it is urgent and vital to establish a correct concept and practicable principles.The authors reviewed recent domestic and international literature and summarized the following treatment procedures and principles for extramural health care of chronic wounds.(1)The patient needs to do self-assessment of the wound by using available simple methods;(2)The patient consults with professional physicians or nurses on wound care to define the severity and etiology of the non-healing wound;(3)Professionals evaluate the existing treatment strategies;(4)Etiological treatments are given by professionals;(5)Patients buy needed dressings via the more convenient ways from pharmacies,e-commerce platform or others;(6)Professionals provide a standardized and reasonable therapeutic plan based on the patient's wound conditions;(7)Both professionals and the patient pay attention to complications to prevent adverse outcomes;(8)Professionals strengthen the public education on wound care and integrated rehabilitation.This review expected to provide new perspectives on the therapeutic strategies for chronic wounds in an extramural setting.

Effect of Guizhi Fuling Capsule on Apoptosis of Myeloma Cells Through Mitochondrial Apoptosis Pathway

Objective: To observe the effects of Guizhi Fuling Capsule(GZFLC) on myeloma cells and explore the mechanisms. Methods: MM1S and RPMI 8226 cells were co-cultured with different concentrations of serum and the cell experiments were divided into negative(10%, 20% and 40%) groups, GZFLC(10%, 20%, and 40%)groups and a control group. Cell counting kit-8(CCK-8) assays and flow cytometry were used to detect the viability and apoptosis levels of myeloma cells. The effects on mitochondria were examined by reactive oxygen specie(ROS) and tetrechloro-tetraethylbenzimidazol carbocyanine iodide(JC-1) assays. Western blot was used to detect the expression of B cell lymphoma-2(Bcl-2), Bcl-2-associated X(Bax), cleaved caspase-3,-9, cytochrome C(Cytc)and apoptotic protease-activating factor 1(Apaf-1). RPMI 8226 cells(2×107) were subcutaneously inoculated into 48 nude mice to study the in vivo antitumor effects of GZFLC. The mice were randomly divided into four groups using a completely randomized design, the high-, medium-, or low-dose GZFLC(840, 420, or 210 mg/kg per day,respectively) or an equal volume of distilled water, administered daily for 15 days. The tumor volume changes in and survival times of the mice in the GZFLC-administered groups and a control group were observed. Cytc and Apaf-1 expression levels were detected by immunohistochemistry. Results: GZFLC drug serum decreased the viability and increased the apoptosis of myeloam cells(P<0.05). In addition, this drug increased the ROS levels and decreased the mitochondrial membrane potential(P<0.01). Western blot showed that the Bcl-2/Bax ratios were decreased in the GZFLC drug serum-treated groups, whereas the expression levels of cleaved caspase-3,-9,Cytc and Apaf-1 were increased(all P<0.01). Over time, the myeloma tumor volumes of the mice in the GZFLCadministered groups decreased, and survival time of the mice in the GZFLC-administered groups were longer than that of the mice in the control group. Immunohistochemical analysis of tumor tissues from the mice in the GZFLCadministered groups revealed that the Cytc and Apaf-1 expression levels were increased(P<0.05). Conclusion:GZFLC promoted apoptosis of myeloma cells through the mitochondrial apoptosis pathway and significantly reduced the tumor volumes in mice with myeloma, which prolonged the survival times of the mice.