Human Serum Albumin Mediated Controllable Synthesis of Defect-Rich Copper Hydroxide Nanowire for Cuproptosis-Based Anti-Tumor Therapy.

Cuproptosis, as a newly identified form of programmed cell death, shows great promise in cancer treatment. Efficient Cu+ delivery while avoiding systemic toxicity and elimination of the resistance from over-expressed intracellular copper chelator glutathione (GSH) are critical for cuproptosis. Herein, this work innovatively constructs a biocompatible and defect-rich copper hydroxide nanowire (HCu nanowire) through a human serum albumin (HSA) mediated biomineralization method. This work finds that the morphology and size of HCu nanowires can be controlled adjusted by the feed ratio of HSA and Cu2+. Remarkably, except for outstanding biocompatibility, HSA coordination endows HCu nanowires abundant oxygen vacancies (OVs), and the defect-rich HCu nanowire possesses excellent GSH consumption efficiency. Density functional theory studies indicate that OVs change GSH absorption energy on defective HCu nanowires. In cancer cells, HCu nanowires deplete GSH and simultaneously produce sufficient free Cu+ for enhanced cuproptosis. Meanwhile, Cu+ can catalyze endogenous H2O2 into hydroxyl radicals (·OH) via a Fenton-like reaction. Thus, synergetic cuproptosis and ROS mediated apoptosis against tumor are achieved. The experimental results show that HCu nanowires have a better performance in both antitumor efficiency and safety compared with chemotherapeutic drug Dox at the same dose, demonstrating its great potential in clinical applications.

Near-Infrared Triggered Self-Accelerating Nanozyme Camouflaged with a Cancer Cell Membrane for Precise Targeted Imaging and Enhanced Cancer Immunotherapy.

Although cancer immunotherapy has made encouraging progress, clinical therapeutic efficiency is often modest due to inadequate immunogenicity and immune resistance. Developing promising nanoagents for simultaneously activating tumor-specific immunity and suppressing immune resistance to achieve efficient immunotherapy is still challenging. Herein, we developed a biomimetic nanozyme consisting of a gold nanorod@mesoporous ceria core-shell scaffold with gold nanoparticle deposition and cancer cell membrane camouflage. The nanozyme exhibited near-infrared (NIR)-enhanced GOx-mimicking activity at high temperatures and performed well under hypoxic environments due to an increased in situ oxygen supply. In cancer cells, the nanozyme induced and amplified hyperthermia by triggering self-accelerating cascade reactions to deplete glucose and inhibiting the expression of heat shock protein under NIR irradiation, which can cause mitochondrial dysfunction and redox balance disruption to activate pyroptosis and elicit a robust immune response. Additionally, the immune checkpoint blockade caused by encapsulated JQ1-mediated PD-L1 downregulation synergistically contributed to excellent immune therapeutic effects. Besides, we demonstrated that cancer cell membrane coating endows the nanozyme targeting ability to tumor. The proposed nanozyme will broaden the application of GOx and have the potential as the nanoplatform for imaging-guided and O2-consuming combined treatments.

An efficient biomimetic nano-regulator inducing simultaneous calcium ion/nitric oxide/energy metabolism triple homeostasis disruption for synergetic cancer therapy.

Inducing homeostasis disruption in cancer cells will cause severe cytotoxicity and apoptosis for cancer therapy. However, comprehensive intracellular active homeostatic mechanisms make it still a daunting challenge. Reported herein is a biomimetic nano-regulator for mutually reinforcing Ca2+/NO/energy metabolism triple homeostasis disruption via cascade reactions.

Effects of the different Tai Chi exercise cycles on patients with essential hypertension: A systematic review and meta-analysis.

Objective: The main treatment for essential hypertension at this stage is pharmacotherapy. Long-term pharmacotherapy is costly with some side effects. Tai Chi, a bright star in traditional Chinese arts, relaxes both mind and body and has been shown to relax blood vessels and lower blood pressure. This study aimed to systematically review the therapeutic effectiveness of the Tai Chi exercise cycle on blood pressure and cardiovascular risk factors of patients with essential hypertension. Methods: Searching CNKI, VIP, CBM, PubMed, EBSCO, Embase, Cochrane Library, and Web of Science to collect randomized controlled trials about Tai Chi exercise in the treatment of patients with essential hypertension according to the inclusion and exclusion criteria. The search time ranged from the date of database construction to December 2022. The Cochrane risk-of-bias tool was used to evaluate the included trials. The meta-analysis was performed with RevMan5.3 and Stata12.0 software. Results: According to the meta-analysis, compared with the controls, Tai Chi exercise with a cycle of more than 12 weeks may be better for the reduction of systolic blood pressure (SBP) [MD = -11.72, 95% CI (-15.52, -7.91)] and diastolic blood pressure (DBP) [MD = -4.68, 95% CI (-7.23, -2.12)], as well as increasing the content of nitric oxide (NO) [MD = 0.99, 95% CI (0.69, 1.28)]. The blood lipid metabolism ability may also be improved after more than 12 weeks of Tai Chi exercise, total cholesterol (TC) [SMD = -0.68, 95% CI (-0.89, -0.46), triglyceride (TG) [SMD = -0.84, 95% CI (-1.25, -0.43)], low-density lipoprotein cholesterol (LDL-C) [SMD = -1.58, 95% CI (-2.29, -0.86)]. However, the improvement of high-density lipoprotein cholesterol (HDL-C) [SMD = 0.54, 95% CI (0.28, 0.79)] was better with a less than 12 weeks exercise cycle. A subgroup analysis for exercise frequency and time showed that the exercise frequency should preferably be more than or equal to 5 times per week for patients with hypertension, and for patients with hypertension plus hyperlipidemia, the exercise frequency less than 5 times per week with exercise time less than 60 min each day may be more beneficial. Conclusion: The meta-analysis indicated that a more than 12 weeks Tai Chi exercise cycle with less than 60 min each time and more than 5 times per week may be more beneficial in blood pressure reduction, NO level increasing and blood lipid metabolism improving in the comparison with the other exercise cycles. For patients with hypertension plus hyperlipidemia, exercise frequency of less than 5 times per week may be better. Systematic Review Registration: [http://www.crd.york.ac.uk/prospero], identifier [CRD42022352035].

Efficacy of Pilates on Pain, Functional Disorders and Quality of Life in Patients with Chronic Low Back Pain: A Systematic Review and Meta-Analysis.

BACKGROUND: Chronic low back pain (CLBP) is a common health problem. Pilates is a unique exercise therapy. This meta-analysis aims to evaluate the efficacy of Pilates on pain, functional disorders, and quality of life in patients with chronic low back pain (CLBP). METHODS: PubMed, Web of Science, CNKI, VIP, Wanfang Data, CBM, EBSCO, and Embase were searched. Randomized controlled trials of Pilates in the treatment of CLBP were collected based on the inclusion and exclusion criteria. The meta-analysis was performed using RevMan 5.4 and Stata 12.2. RESULTS: 19 randomized controlled trials with a total of 1108 patients were included. Compared with the controls, the results showed the following values: Pain Scale [standard mean difference; SMD = -1.31, 95%CI (-1.80, -0.83), p < 0.00001], Oswestry Disability Index (ODI) [mean difference; MD = -4.35, 95%CI (-5.77, -2.94), p < 0.00001], Roland-Morris Disability Questionnaire (RMDQ) [MD = -2.26, 95%CI (-4.45, -0.08), p = 0.04], 36-item Short-Form (SF-36) (Physical Function (PF) [MD = 5.09, 95%CI (0.20, 9.99), p = 0.04], Role Physical (RP) [MD = 5.02, 95%CI (-1.03, 11.06), p = 0.10], Bodily Pain (BP) [MD = 8.79, 95%CI (-1.57, 19.16), p = 0.10], General Health (GH) [MD = 8.45, 95%CI (-5.61, 22.51), p = 0.24], Vitality (VT) [MD = 8.20, 95%CI(-2.30, 18.71), p = 0.13], Social Functioning (SF) [MD = -1.11, 95%CI (-7.70, 5.48), p = 0.74], Role Emotional (RE) [MD = 0.86, 95%CI (-5.53, 7.25), p = 0.79], Mental Health (MH) [MD = 11.04, 95%CI (-12.51, 34.59), p = 0.36]), Quebec Back in Disability Scale (QBPDS) [MD = -5.51, 95%CI (-23.84, 12.81), p = 0.56], and the sit-and-reach test [MD = 1.81, 95%CI (-0.25, 3.88), p = 0.09]. CONCLUSIONS: This meta-analysis reveals that Pilates may have positive efficacy for pain relief and the improvement of functional disorders in CLBP patients, but the improvement in quality of life seems to be less obvious. REGISTRATION: PROSPERO CRD42022348173.

Harnessing Reconstructed Macrophage Modulation of Infiltration-Excluded Immune Microenvironments To Delineate Glioma Infiltrative Region.

High invasiveness of glioma produces residual glioma cells in the brain parenchyma after surgery and ultimately causes recurrence. Precise delineation of glioma infiltrative region is critical for an accurate complete resection, which is challenging. The glioma-infiltrating area constitutes infiltration-excluded immune microenvironments (I-E TIMEs), which recruits endogenous or adoptive macrophages to the invasive edge of glioma. Thus, combined with immune cell tracing technology, we provided a novel strategy for the preoperative precise definition of the glioma infiltration boundary, even satellite-like infiltration stoves. Herein, the biomimetic probe was constructed by internalizing fluorophore labeled PEGylated KMnF3 nanoparticles into bone-marrow-derived macrophages using magnetic resonance imaging (MRI)/fluorescence imaging (FI). The biomimetic probe was able to cross the blood-brain barrier and home to the orthotopic glioma infiltrates including satellite stove under MRI and FI tracing, which was validated using hematoxylin and eosin staining, indicating its excellent performance in distinguishing the margins between the glioma cell and normal tissues. This study guides the precise definition of glioma infiltration boundaries at the cellular level, including the observation of any residual glioma cells after surgery. Thus, it has the potential to guide surgery to maximize resection and predict recurrence in the clinic.

Effectiveness of the Rehabilitation Training Combined with Maitland Mobilization for the Treatment of Chronic Ankle Instability: A Randomized Controlled Trial.

The study aims to determine whether routine rehabilitation training combined with the Maitland mobilization is more effective than routine rehabilitation training alone in patients with chronic ankle instability, intending to provide a novel rehabilitation strategy for chronic ankle instability. A total of 48 subjects were divided into three groups: EG (Maitland mobilization and routine rehabilitation), CG (routine rehabilitation), and SG (sham mobilization and routine rehabilitation). The intervention was performed three times each week for 4 weeks, for a total of 12 sessions. Before and after the intervention, the muscle strength, star excursion balance test (SEBT), weight-bearing dorsiflexion range of motion (WB-DFROM), ankle range of movement, Cumberland ankle instability tool (CAIT), self-comfort visual analog scale (SCS-VAS), and self-induced stability scale (SISS-VAS) were assessed. The results showed that the improvement of SEBT, WB-DFROM, and active ankle range of movement without the pain in EG was more obvious than CG and SG, but the improvement of the self-report of ankle severity and muscle strength was not. Compared with routine rehabilitation training alone, routine rehabilitation training combined with Maitland mobilization for patients with chronic ankle instability may provide more benefit in terms of balance and ankle range of movement than routine rehabilitation alone, but the improvement in muscle strength was not evident enough.

Does whole-body vibration training have a positive effect on balance and walking function in patients with stroke? A meta-analysis.

Objective: After a stroke, patients usually suffer from dysfunction, such as decreased balance ability, and abnormal walking function. Whole-body vibration training can promote muscle contraction, stimulate the proprioceptive system, enhance the muscle strength of low limbs and improve motor control ability. The study aims to evaluate the effectiveness of whole-body vibration training on the balance and walking function of patients with stroke. Methods: PubMed, CNKI, VIP, CBM, EBSCO, Embase and Web of Science were searched. According to the inclusion and exclusion criteria, randomized controlled trials on the effectiveness of whole-body vibration training on the balance and walking function of patients with stroke were collected. The search time ranged from the date of database construction to November 2022. The included trials were evaluated by the Cochrane risk-of-bias tool. The meta-analysis was performed using two software packages, consisting of RevMan 5.4 and Stata 12.2. If the results included in the literature were continuous variables, use the mean difference (MD) and 95% confidence interval (CI) for statistics. Results: (1) A total of 22 randomized controlled trials (RCTs) with a total of 1089 patients were included. (2) The results of meta-analysis showed that: compared with the controls, step length (MD = 6.12, 95%CI [5.63, 6.62], p < 0.001), step speed (MD = 0.14, 95%CI [0.09, 0.20], p < 0.001), cadence (MD = 9.03, 95%CI [2.23, 15.83], p = 0.009), stride length (MD = 6.74, 95%CI [-3.47, 10.01], p < 0.001), Berg Balance Scale (BBS) (MD = 4.08, 95%CI [2.39, 5.76], p < 0.001), Timed Up-and-Go test (TUGT) (MD = -2.88, 95%CI [-4.94, 0.81], p = 0.006), 10-meter Walk Test (10MWT) (MD = -2.69, 95%CI [-3.35, -2.03], p < 0.001), functional ambulation category scale (FAC) (MD = 0.78, 95%CI [0.65, 0.91], p < 0.001), Fugl-Meyer motor assessment of lower extremity (FMA-LE) (MD = 4.10, 95%CI [2.01, 6.20], p = 0.0001). (3) The results of subgroup analysis showed that, compared with other vibration frequencies, at 20-30 Hz frequency, WBV training had an obvious improvement effect only in TUGT. (4) The safety analysis showed that WBV training may be safe. Conclusion: Whole-body vibration training has a positive effect on the balance and walking function of patients with stroke. Thus, whole-body vibration training is a safe treatment method to improve the motor dysfunction of patients with stroke. Systematic review registration: [http://www.crd.york.ac.uk/PROSPERO], identifier [CRD4202348263].

Functionalized nanoprobes for in situ detection of telomerase.

Telomerase, a special ribonucleoprotein reverse transcriptase, can maintain the length and stability of telomeres and plays an important role in cell proliferation and differentiation. Due to the distinguishable expression level in normal cells and cancer cells, telomerase has become an important biomarker for cancer diagnosis and prognosis evaluation. Despite major breakthroughs in the field of telomerase detection, the extracts in the cell lysate are still the first choice as the analyte nevertheless, which will bring serious inaccuracies compared with the real intracellular activity. With the development of nanotechnology and nanomaterials, extraordinary progress has been made in telomerase detection by employing different versatile nanoprobes. In this review, we list the superiority of nanoprobes and systematically summarize the applications of nanoprobes in telomerase detection from the aspects of various nanomaterials and discuss the current challenges and potential trends in the future design of nanoprobes.

Nanoenzymes in disease diagnosis and therapy.

Nanoenzymes, as a new generation of artificial enzymes with highly effective enzyme-like properties, have come into focus and have achieved great progress. Considering the enormous potential of nanoenzymes in the field of disease diagnosis and treatment, such as biological detection, anti-inflammatory, anti-oxidative damage and cancer treatments, etc., we firstly summarize the wide range of bio-applications of different nanoenzymes in these fields. In this review, we discuss the factors influencing the properties of nanoenzymes such as pH and temperature, surface modification and doping of other elements. In addition, we introduce five kinds of nanoenzymes and their bio-applications in various diseases. The current review will make researchers better understand the current state of the development of nanoenzymes and may be beneficial for new breakthroughs.

A cancer cell membrane-encapsulated MnO2 nanoreactor for combined photodynamic-starvation therapy.

We demonstrate a MnO2-based nanoreactor to achieve continuous oxygen generation and efficient conversion from glucose to singlet oxygen for combined photodynamic-starvation therapy.

A biomimetic nanoreactor for synergistic chemiexcited photodynamic therapy and starvation therapy against tumor metastasis.

Photodynamic therapy (PDT) is ineffective against deeply seated metastatic tumors due to poor penetration of the excitation light. Herein, we developed a biomimetic nanoreactor (bio-NR) to achieve synergistic chemiexcited photodynamic-starvation therapy against tumor metastasis. Photosensitizers on the hollow mesoporous silica nanoparticles (HMSNs) are excited by chemical energy in situ of the deep metastatic tumor to generate singlet oxygen (1O2) for PDT, and glucose oxidase (GOx) catalyzes glucose into hydrogen peroxide (H2O2). Remarkably, this process not only blocks the nutrient supply for starvation therapy but also provides H2O2 to synergistically enhance PDT. Cancer cell membrane coating endows the nanoparticle with biological properties of homologous adhesion and immune escape. Thus, bio-NRs can effectively convert the glucose into 1O2 in metastatic tumors. The excellent therapeutic effects of bio-NRs in vitro and in vivo indicate their great potential for cancer metastasis therapy.

A graphene-based fluorescent nanoprobe for simultaneous monitoring of miRNA and mRNA in living cells.

Since microRNA-21 (miR-21) and protein programmed cell death 4 (PDCD4) are implicated in tumor progression, simultaneous monitoring of the dynamic variation of miR-21 and PDCD4 mRNA in living cells is of great interest for understanding their relationship in the pathology of the disease. Herein, a versatile nanoprobe based on graphene oxide (GO) and DNA was prepared which can simultaneously monitor and visualize miR-21 and PDCD4 mRNA in living cells. In vitro experiments demonstrate that the nanoprobe exhibits outstanding selectivity and high sensitivity towards miR-21 and PDCD4 mRNA. Moreover, by exploiting the DNA/GO sensing platform, simultaneous imaging of miR-21 and PDCD4 mRNA in MCF-7 human breast cancer cells has been realized. More significantly, an inverse correlation between miR-21 and PDCD4 mRNA was verified by fluorescence confocal imaging and RT-PCR results, indicating the distinguishing ability of the nanoprobe at various expression levels of RNAs. The current strategy provides a promising tool to implement the detection of miR-21 and PDCD4 mRNA in cancer cells and can be expected to be applied for other biomolecular sensing.

Nuclear-Targeted Photothermal Therapy Prevents Cancer Recurrence with Near-Infrared Triggered Copper Sulfide Nanoparticles.

Clinical cancer treatments nowadays still face the challenge of recurrence due to the residual cancer cells and minute lesions in surgeries or chemotherapies. To effectively address the problem, we introduce a strategy for constructing cancer cell nuclear-targeted copper sulfide nanoparticles (NPs) with a significant photothermal effect to completely kill residual cancer cells and prevent local cancer recurrence. The NPs could directly target the tumor cells and further enter the nucleus by the surface modification of RGD and TAT peptides. Under the irradiation of 980 nm near-infrared laser, the NPs rapidly increase the temperature of the nucleus, destroy the genetic substances, and ultimately lead to an exhaustive apoptosis of the cancer cells. In vivo experiments show that the designed NPs could effectively treat cancer and prevent the return of cancer with a single laser irradiation for 5 min. The photothermal therapy strategy with nuclear targeting for cancer therapy and anti-recurrence will provide more possibilities to develop efficient platforms for treating cancer.

A pre-protective strategy for precise tumor targeting and efficient photodynamic therapy with a switchable DNA/upconversion nanocomposite.

Tumor-specific targeting based on folic acid (FA) is one of the most common and significant approaches in cancer therapy. However, the expression of folate receptors (FRs) in normal tissues will lead to unexpected targeting and unsatisfactory therapeutic effect. To address this issue, we develop a pre-protective strategy for precise tumor targeting and efficient photodynamic therapy (PDT) using a switchable DNA/upconversion nanocomposite, which can be triggered in the acidic tumor microenvironment. The DNA/upconversion nanocomposite is composed of polyacrylic acid (PAA) coated upconversion nanoparticles (UCNPs), the surface of which is modified using FA and chlorin e6 (Ce6) functionalized DNA sequences with different lengths. Initially, FA on the shorter DNA was protected by a longer DNA to prevent the bonding to FRs on normal cells. Once reaching the acidic tumor microenvironment, C base-rich longer DNA forms a C-quadruplex, resulting in the exposure of the FA groups and the bonding of FA and FRs on cancer cell membranes to achieve precise targeting. Simultaneously, the photosensitizer chlorin e6 (Ce6) gets close to the surface of UCNPs, enabling the excitation of Ce6 to generate singlet oxygen (1O2) under near infrared light via Förster resonance energy transfer (FRET). In vivo experiments indicated that higher tumor targeting efficiency was achieved and the tumor growth was greatly inhibited through the pre-protective strategy.

Reversing Multidrug Resistance by Multiplexed Gene Silencing for Enhanced Breast Cancer Chemotherapy.

Multidrug resistance (MDR), as one of the main problems in clinical breast cancer chemotherapy, is closely related with the over expression of drug efflux transporter P-glycoprotein (P-gp). In this study, a novel drug-loaded nanosystem was developed for inhibiting the P-gp expression and reversing MDR by multiplexed gene silencing, which composes of graphene oxide (GO) modified with two molecular beacons (MBs) and Doxorubicin (Dox). When the nanosystem was uptaken by the MDR breast cancer cells, Dox was released in the acidic endosomes and MBs were hybridized with target sequences. The intracellular multidrug resistance 1 (MDR1) mRNA and upstream erythroblastosis virus E26 oncogene homolog 1 (ETS1) mRNA can be silenced by MBs, which can effectively inhibit the expression of P-gp and further prevent the efflux of Dox and reverse MDR. In vitro and in vivo studies indicated that the strategy of reversing MDR by multiplexed gene silencing could obviously increase MCF-7/Adr cells' Dox accumulation and enormously enhance the therapeutic efficacy of MDR breast cancer chemotherapy.

A simple approach for glutathione functionalized persistent luminescence nanoparticles as versatile platforms for multiple in vivo applications.

We develop a simple method by constructing glutathione (GSH) conjugated persistent luminescence nanoparticles (PLNPs-GSH) as versatile platforms for multiple biological applications. PLNPs-GSH possess enhanced water solubility and contains a large number of active groups, which offer opportunities for further modification with different functional groups.

Nanocarriers with multi-locked DNA valves targeting intracellular tumor-related mRNAs for controlled drug release.

The fabrication of well-behaved drug delivery systems that can transport drugs to specifically treat cancer cells rather than normal cells is still a tremendous challenge. A novel drug delivery system with two types of tumor-related mRNAs as "keys" to open the multiple valves of the nanocarrier to control drug release was developed. Hollow mesoporous silica nanoparticles were employed as the nanocarrier and dual DNAs targeting two intracellular mRNAs were employed as "multi-locks" to lock up the nanocarrier. When the nanocarrier enters the cancer cells, the overexpressed endogenous mRNA keys hybridize with the DNA multi-locks to open the valves and release the drug. Each single mRNA could not trigger the opening of the locks to release the cargo. Therefore, the nanocarrier can be applied for specific chemotherapy against cancer cells with minor side effects to normal cells. The current strategy could provide an important avenue towards advancing the practical applications of drug delivery systems used for cancer therapy.

Tumor microenvironment-triggered fabrication of gold nanomachines for tumor-specific photoacoustic imaging and photothermal therapy.

Nanoparticles as novel theranostic agents for cancer treatment have been extensively investigated in recent years. However, the poor tumor selectivity and retention of the theranostic agents result in unsatisfactory performance of both the diagnostic and therapeutic functions. Herein, we developed an alpha-cyclodextrin (α-CD)-based gold/DNA nanomachine for tumor-selective diagnosis and therapy. The α-CDs were capped at the ends of DNA, and their release was triggered by the low pH of the tumor microenvironment, which further resulted in DNA self-assembly through complementary base pairing. The large-sized gold aggregates failed to escape from the tumor tissue, thereby realizing the goal of tumor-specific targeting and enhanced retention. Thus, the photoacoustic signal and photothermal effect are also activated, thereby achieving tumor-targeted photoacoustic imaging and photothermal therapy. In vivo results indicated that the designed gold nanomachines can serve as efficient theranostic agents for diagnosis and therapy. Moreover, we found that the α-CD caps have the ability to protect the nanoparticles from clearance and enzyme digestion, which helps the nanoparticles reach the tumor more efficiently.

Dual-Ratiometric Fluorescent Nanoprobe for Visualizing the Dynamic Process of pH and Superoxide Anion Changes in Autophagy and Apoptosis.

Autophagy and apoptosis are closely associated with various pathological and physiological processes in cell cycles. Investigating the dynamic changes of intracellular active molecules in autophagy and apoptosis is of great significance for clarifying their inter-relationship and regulating mechanism in many diseases. In this study, we develop a dual-ratiometric fluorescent nanoprobe for quantitatively differentiating the dynamic process of superoxide anion (O2•-) and pH changes in autophagy and apoptosis in HeLa cells. A rhodamine B-loaded mesoporous silica core was used as the reference, and fluorescence probes for pH and O2•- measurement were doped in the outer layer shell of SiO2. Then, chitosan and triphenylphosphonium were modified on the surface of SiO2. The experimental results showed that the nanoprobe is able to simultaneously and precisely visualize the changes of mitochondrial O2•- and pH in HeLa cells. The kinetics data revealed that the changes of pH and O2•- during autophagy and apoptosis in HeLa cells were significantly different. The pH value was decreased at the early stage of apoptosis and autophagy, whereas the O2•- level was enhanced at the early stage of apoptosis and almost unchanged at the initial stage of autophagy. At the late stage of apoptosis and autophagy, the concentration of O2•- was increased, whereas the pH was decreased at the late stage of autophagy and almost unchanged at the late stage of apoptosis. We hope that the present results provide useful information for studying the effects of O2•- and pH in autophagy and apoptosis in various pathological conditions and diseases.