Influence of Different Ratios of DSPE-PEG2k on Ester Prodrug Self-Assembly Nanoparticles for Cell Migration and Proliferation Suppression.

Background: Bufalin (BFL, an active anti-tumor compound derived from toad venom) is limited in its application due to high toxicity and rapid metabolism of the cardiotonic steroid. Ester prodrug self-assembly nanoparticles have shown significant improved effects in addressing the above-mentioned issues. Methods: An ester bond was formed between linoleic acid and bufalin to synthesize linoleic acid-bufalin prodrug (LeB). The self-assembly nanoparticles (LeB-PSNs) containing different mass ratios of DSPE-PEG2k and prodrug (6:4, 7:3, 8:2, 9:1 and 10:0) were prepared via co-precipitation method and defined as 6:4-PSNs, 7:3-PSNs, 8:2-PSNs, 9:1-PSNs and LeB-PSNs, respectively. Further, the characterization (particle size, zeta potential, surface morphology and stability) of the nanoparticles was carried out. Finally, we evaluated the impact of different ratios of DSPE-PEG2k on the hydrolysis rate, cytotoxicity, cellular uptake, cell migration and proliferation suppression potential of the prodrug nanoparticles. Results: The linoleic acid-bufalin prodrug (LeB) was successfully synthesized. Upon the addition of DSPE-PEG2k at different weight ratios, both particle size and polydispersity index (PDI) significantly decreased, while the zeta potential increased remarkably. No significant differences in particle size, PDI and Zeta potential were observed among the 9:1, 8:2 and 7:3 PSNs. Notably, the 8:2 (w/w) DSPE-PEG2k nanoparticles exhibited superior stability, hydrolysis and cellular uptake rates, along with efficient cell cytotoxicity, cell migration and proliferation suppression. Conclusion: These findings indicate that DSPE-PEG2k could improve the performance of BFL prodrug nanoparticles, namely enhancing stability and achieving adaptive drug release by modulating the hydrolysis rate of esterase. This study therefore provides more opportunities for the development of BFL application.

Improving cellular uptake and synergetic anti-tumor effects of magnolol and Brucea javanica oil through self-microemulsion.

OBJECTIVE: Magnolol (MG) and Brucea javanica (L.) Merr. oil (BJO) possess synergetic anti-tumor effects, but have poor water solubility and stability, which results in low oral bioavailability. SIGNIFICANCE: The MG loaded self-microemulsion drug delivery system (MG-SMDDS) with BJO as oil phase component was utilized to improve the cellular uptake and synergetic anti-tumor effects. METHODS: Compatibility study and pseudoternary phase diagram (PTPD) were respectively employed to screen for the composition and proportion of oil phase in the formulation. Central composite design-effect surface method was applied to optimize proportion of each formulation condition. The droplet size, ζ-potential, colloid stability, encapsulation rate (ER) and in vitro dissolution rate of MG-SMDDS were evaluated. Furthermore, cellular uptake and cytotoxicity of the microemulsion on HepG2 cells were assessed. RESULTS: The optimal composition of MG-SMDDS was: MG (9.09%), castor oil (7.40%), BJO (2.47%), Cremophor EL 35 (54.04%) and 1, 2-propanediol (27.01%). The MG-SMDDS exhibited satisfactory droplet size, ζ-potential, colloid stability and ER, as well as faster dissolution rate than free MG. More importantly, SMEDDS containing BJO could enhance the cellular uptake and cytotoxicity of free BJO and free MG on tumor cells. CONCLUSIONS: The BJO self-microemulsion delivery technique can provide an idea for design of oral delivery vehicles based on BJO.

Intrafraction Motion and Margin Assessment for Ethos Online Adaptive Radiotherapy Treatments of the Prostate and Seminal Vesicles.

Purpose: Online adaptive radiation therapy (OART) uses daily imaging to identify changes in the patient's anatomy and generate a new treatment plan adapted to these changes for each fraction. The aim of this study was to determine the intrafraction motion and planning target volume (PTV) margins required for an OART workflow on the Varian Ethos system. Methods and Materials: Sixty-five fractions from 13 previously treated OART patients were analyzed for this retrospective study. The prostate and seminal vesicles were contoured by a radiation oncologist on 2 cone beam computed tomography scans (CBCT) for each fraction, the initial CBCT at the start of the treatment session, and the verification CBCT immediately before beam-on. In part 1 of the study, PTVs of different sizes were defined on the initial CBCT, and the geometric overlap with the clinical target volume (CTV) on the verification CBCT was used to determine the optimal OART margin. This was performed with and without a patient realignment shift by registering the verification CBCT to the initial CBCT. In part 2 of the study, the margins determined in part 1 were used for simulated Ethos OART treatments on all 65 fractions. The resultant coverage to the CTV on the verification CBCT, was compared with an image guided radiation therapy (IGRT) workflow with 7-mm margins. Results: Part 1 of the study found, if a verification CBCT and shift is performed, a 4-mm margin on the prostate and 5 mm on the seminal vesicles resulted in 95% of the CTV covered by the PTV in >90% of fractions, and 98% of the CTV covered by the PTV in >80% of fractions. Part 2 of the study found when these margins were used in an Ethos OART workflow, they resulted in CTV coverage that was superior to an IGRT workflow with 7-mm margins. Conclusions: A 4mm prostate margin and 5-mm seminal vesicles margin in an OART workflow with verification imaging are adequate to ensure coverage on the Varian Ethos system. Larger margins may be required if using an OART workflow without verification imaging.

Management and clinical outcomes of follicular lymphoma across continuous lines of treatments: a retrospective analysis in China.

Background: Follicular lymphoma (FL) is characterized by an incurable course that frequently necessitates multiple lines of treatment. While a range of new approaches have broadened therapeutic options for patients in later lines, data regarding treatment patterns and outcomes of Chinese patients with relapsed/refractory(R/R) FL was scarcely reported. Methods: This retrospective single-center study included patients diagnosed with FL grades 1-3a at our institution between January 2002 and December 2019. Endpoints of interest were analyzed according to lines and types of interventions. The endpoints mainly included overall response rate (ORR), progression-free survival (PFS), and overall survival (OS). Results: The study enrolled 566 biopsy-proven patients. Among them, 544 patients initiated the first line of treatment, followed by 240 initiating the second line, 146 initiating the third line, 88 initiating the fourth line, 47 initiating the fifth line, and 28 initiating the sixth line. In terms of treatment patterns, anti-CD20 chemotherapy was a major modality in the first and second lines. However, for patients in the third line and subsequent lines, treatment approaches were diverse, and participation in clinical trials for new medications was common, which correlated with a survival benefit. The study also revealed that clinical indicators (such as ORR, PFS, and OS) gradually decreased with each subsequent line of treatment. The ORR at the first line was 86.6%, but decreased to 48.6% at the third line and 40.4% at the sixth line, respectively. Similarly, median OS and PFS decreased to 88.8 and 7.1 months at the third line and further reduced to 21.7 and 2.8 months at the sixth line, respectively. A total of 133 patients developed progression within 24 months from the initiation of first line anti-CD20 chemotherapy (POD24), and these patients exhibited poorer response rates and outcomes in subsequent lines of therapycompared to the non-POD24 group. Conclusion: This study revealed the clinical routine practices and prognosis of R/R FL patients within the Chinese population. It underscored the unmet need for optimal strategies to improve survival and also served as a benchmark for future trials.

Design, Synthesis, and Antitumor Efficacy of Substituted 2-Amino[1,2,4]triazolopyrimidines and Related Heterocycles as Dual Inhibitors for Microtubule Polymerization and Janus Kinase 2.

Preclinical and clinical studies have demonstrated the synergistic effect of microtubule-targeting agents in combination with Janus kinase 2 (JAK2) inhibitors, prompting the development of single agents with enhanced therapeutic efficacy by dually inhibiting tubulin polymerization and JAK2. Herein, we designed and synthesized a series of substituted 2-amino[1,2,4]triazolopyrimidines and related heterocycles as dual inhibitors for tubulin polymerization and JAK2. Most of these compounds exhibited potent antiproliferative activity against the selected cancer cells, with compound 7g being the most active. This compound effectively inhibits both tubulin assembly and JAK2 activity. Furthermore, phosphorylated compound 7g (i.e., compound 7g-P) could efficiently convert to compound 7g in vivo. Compound 7g, whether it was administered directly or in the form of a phosphorylated prodrug (i.e., compound 7g-P), significantly inhibited the growth of A549 xenografts in nude mice. The present findings strongly suggest that compound 7g represents a promising chemotherapeutic agent with high antitumor efficacy.

The Hidden Patient Connections: Predicting Hormonal Therapy Medication Discontinuation Using Hypergraph Neural Network on Clinical Communications.

Hormonal therapy is an important adjuvant treatment for breast cancer patients, but medication discontinuation of such therapy is not uncommon. The goal of this paper is to conduct research on the modeling of clinic communications, which have shown value in understanding medication discontinuation, to predict the discontinuation of hormonal therapy medications. Notably, we leveraged the Hypergraph Neural Network to capture the hidden connections of patients that were inferred from clinical communications. Combining the content of clinical communications as well as the demographics, insurance, and cancer stage information, our model achieved an AUC of 67.9%, which significantly outperformed other baselines such as Graph Convolutional Network (65.3%), Random Forest (62.7%), and Support Vector Machine (62.8%). Our study suggested that incorporating the hidden patient connections encoded in clinical communications into prediction models could boost their performance. Future research would consider combining structured medical records and clinical communications to better predict medication discontinuation.

Clinical assessment of a novel machine-learning automated contouring tool for radiotherapy planning.

Contouring has become an increasingly important aspect of radiotherapy due to inverse planning. Several studies have suggested that the clinical implementation of automated contouring tools can reduce inter-observer variation while increasing contouring efficiency, thereby improving the quality of radiotherapy treatment and reducing the time between simulation and treatment. In this study, a novel, commercial automated contouring tool based on machine learning, the AI-Rad Companion Organs RT™ (AI-Rad) software (Version VA31) (Siemens Healthineers, Munich, Germany), was assessed against both manually delineated contours and another commercially available automated contouring software, Varian Smart Segmentation™ (SS) (Version 16.0) (Varian, Palo Alto, CA, United States). The quality of contours generated by AI-Rad in Head and Neck (H&N), Thorax, Breast, Male Pelvis (Pelvis_M), and Female Pelvis (Pevis_F) anatomical areas was evaluated both quantitatively and qualitatively using several metrics. A timing analysis was subsequently performed to explore potential time savings achieved by AI-Rad. Results showed that most automated contours generated by AI-Rad were not only clinically acceptable and required minimal editing, but also superior in quality to contours generated by SS in multiple structures. In addition, timing analysis favored AI-Rad over manual contouring, indicating the largest time saving (753s per patient) in the Thorax area. AI-Rad was concluded to be a promising automated contouring solution that generated clinically acceptable contours and achieved time savings, thereby greatly benefiting the radiotherapy process.

Design, Synthesis and Antitumor Activity of Novel Selenium-Containing Tepotinib Derivatives as Dual Inhibitors of c-Met and TrxR.

Cellular mesenchymal-epithelial transition factor (c-Met), an oncogenic transmembrane receptor tyrosine kinase (RTK), plays an essential role in cell proliferation during embryo development and liver regeneration. Thioredoxin reductase (TrxR) is overexpressed and constitutively active in most tumors closely related to cancer recurrence. Multi-target-directed ligands (MTDLs) strategy provides a logical approach to drug combinations and would adequately address the pathological complexity of cancer. In this work, we designed and synthesized a series of selenium-containing tepotinib derivatives by means of selenium-based bioisosteric modifications and evaluated their antiproliferative activity. Most of these selenium-containing hybrids exhibited potent dual inhibitory activity toward c-Met and TrxR. Among them, compound 8b was the most active, with an IC50 value of 10 nM against MHCC97H cells. Studies on the mechanism of action revealed that compound 8b triggered cell cycle arrest at the G1 phase and caused ROS accumulations by targeting TrxR, and these effects eventually led to cell apoptosis. These findings strongly suggest that compound 8b serves as a dual inhibitor of c-Met and TrxR, warranting further exploitation for cancer therapy.

Individualized breathing trace quality assurance for lung radiotherapy patients undergoing 4DCT simulation.

4DCT simulation is a popular solution for radiotherapy simulation of lung cancer patients as it allows the clinician to gain an appreciation for target motion during the patient breathing cycle. Resultant binning of images and production of the 4DCT dataset relies heavily on the recorded breathing trace; but quality assurance is not routinely performed on these and there lacks any substantial recommendations thereof. An application was created for Windows in C# that was able to analyze the VXP breathing trace files from Varian RPM/RGSC and quantify various metrics associated with the patient breathing cycle. This data was then used to consider errors in voluming of targets for several example cases in order to justify recommendations on quality assurance. For 281 real patient breathing traces from 4DCT simulation of lung targets, notable differences were found between RGSC and application calculations of phase data. For any new patient without individualized QA, the average marked phase calculation (which is used for 4DCT reconstruction) is only accurate to within 19% of the actual phases. The error in BPM within the scan due to breathing rate variation is 37%. The uncertainty in amplitude due to breathing variation is 34% in the mean. Phase uncertainty leads to misbinning which we have shown can lead to missing 66% of the target for gated treatment. Variation in inhalation/exhalation level leads to voluming errors which, without individualized QA, can be assumed to be 11% (PTV is smaller than actual). Without individualized quality assurance of patient breathing traces, large uncertainties have to be assumed for metrics of both phase and amplitude, leading to clinically significant uncertainties in treatment. It is recommended to perform individualized quality assurance as this provides the clinician with an accurate quantification of uncertainty for their patient.

Assessment of semi-automated stereotactic treatment planning for online adaptive radiotherapy in ethos.

INTRODUCTION: The Ethos treatment planning system allows for the rapid generation of online adaptive treatment plans while the patient is on the treatment couch. One promising application of online adaptive radiotherapy is its use in stereotactic radiotherapy. The purpose of this study was to ensure the Ethos treatment planning system (TPS) can produce clinically acceptable stereotactic plans, that are non-inferior to those from the Eclipse TPS. METHOD: Forty patients that received previous stereotactic radiotherapy treatment on a Halcyon, 20 of which were lung cases, and 20 that were brain cases, were replanned using the Ethos TPS. The generated IMRT and VMAT plans were compared to the clinical Eclipse VMAT plan. RESULTS: This study found that the Ethos TPS can produce VMAT plans of equivalent quality (target coverage, conformity and OAR doses) to those from the Eclipse TPS for lung SBRT and brain SRT. The IMRT plans produced by the Ethos planning system were marginally inferior to Eclipse VMAT plans, with the differences likely primarily due to beam geometry rather than the optimization system. Ethos plans were generally more modulated than Eclipse plans. With careful selection of optimization structures and reduction in the body contour, VMAT plan generation time could be reduced by 87%. CONCLUSION: Ethos can generate stereotactic VMAT plans that are equivalent to those from Eclipse in the timeframe required for online adaptive radiotherapy.

Clinical outcomes of endoscopic versus laparoscopic resection for senile patients with gastric gastrointestinal stromal tumours (2 to 4 cm) originating from the muscularis propria layer.

Introduction: Currently, there still are no selection criteria for endoscopic resection (ER) versus laparoscopic resection (LR) of gastric gastrointestinal stromal tumours (GIST) (2 to 4 cm) originating from the muscularis propria layer (MP-GISTs). Aim: To investigate and compare the long-term prognosis of ER and LR for resecting gastric MP-GISTs, with at least 5 years of follow-up. Material and methods: Between January 2010 and December 2015, 134 patients with gastric MP-GISTs were consecutively enrolled in this study. The main comparison measurements included the short-term and long-term outcomes between the ER group (n = 89) and the LR group (n = 45). Results: In this study, there were no significant differences in the rates of complete resection (p = 0.220) and short-term complications (p = 0.663) between the ER group and the LR group. The ER group had a shorter operation time (50.1 ±18.2 min vs. 120.6 ±32.5 min, p < 0.001), shorter hospital stays (5.1 ±1.9 days vs. 6.4 ±3.7 days, p = 0.026), and lower hospitalization costs (16639.5 ±5091.3 CNY vs. 24030.4 ±6803.1 CNY, p < 0.001) than the LR group. The ER group had a lower rate of long-term complications than the LR group (p = 0.001) during the follow-up period (84.2 ±17.9 months vs. 89.0 ±16.8 months, p = 0.207). Conclusions: Our results showed that ER was a more feasible treatment approach than LR when the gastric MP-GIST was located in or near the cardia/pylorus. ER also had several other advantages over LR, such as a shorter procedure time, shorter hospital stay, and lower hospitalization costs.

Access to 3-Amino-[1,2,4]-triazolo Pyridines and Related Heterocycles via Electrochemically Induced Desulfurative Cyclization.

An efficient, one-pot approach has been established for synthesizing a wide range of 3-amino-[1,2,4]-triazolo pyridines and related heterocycles from the electrochemically induced desulfurative cyclization of 2-hydrazinopyridines with isothiocyanates. The protocol allows for the formation of C-N bonds under simple conditions without transition metals or external oxidants. The practicability of this strategy is demonstrated by its broad substrate scope, good functional group compatibility, and gram-scale synthesis. The late-stage modification of 3-amino-[1,2,4]-triazolo pyridines enables us to obtain several molecules with potent anticancer activity.

Characterization of an advanced cone beam CT (CBCT) reconstruction algorithm used for dose calculation on Varian Halcyon linear accelerators.

In this study, the performance of a new iterative reconstruction algorithm, the pre-clinical AcurosXB iCBCT algorithm, has been characterized on Varian Halcyon linear accelerators with respect to the potential of radiotherapy dose calculations on CBCT images. The study utilized various phantom setups to verify the accuracy of the pre-clinical algorithm under different scatter conditions and compared dose calculations performed on CBCT images reconstructed with the pre-clinical algorithm to those performed on typical planning CT images. The results indicated that despite showing improvements compared to the existing iCBCT protocol, certain restrictions should be introduced when the pre-clinical AcurosXB iCBCT algorithm was used for dose calculations. Changes in the scatter condition exhibited a larger effect on CBCTs than on planning CTs. Therefore, users should be careful in offsetting the patient and positioning the patient's arms if the resultant images will be used for dose calculations. In addition, protocols with different kV settings should be approached with caution, where 100 kV protocols should only be used to scan the head and neck area, while the rest of the body should be scanned with the 125 kV and 140 kV protocols. When the patient is set up properly and the appropriate energy is selected for the anatomical area, the uncertainty of using the novel AcurosXB iCBCT algorithm for treatment planning dose calculation is within ±2.0%.

Varian ethos online adaptive radiotherapy for prostate cancer: Early results of contouring accuracy, treatment plan quality, and treatment time.

The Varian Ethos system allows for online adaptive treatments through the utilization of artificial intelligence (AI) and deformable image registration which automates large parts of the anatomical contouring and plan optimization process. In this study, treatments of intact prostate and prostate bed, with and without nodes, were simulated for 182 online adaptive fractions, and then a further 184 clinical fractions were delivered on the Ethos system. Frequency and magnitude of contour edits were recorded, as well as a range of plan quality metrics. From the fractions analyzed, 11% of AI generated contours, known as influencer contours, required no change, and 81% required minor edits in any given fraction. The frequency of target and noninfluencer organs at risk (OAR) contour editing varied substantially between different targets and noninfluencer OARs, although across all targets 72% of cases required no edits. The adaptive plan was the preference in 95% of fractions. The adaptive plan met more goals than the scheduled plan in 78% of fractions, while in 15% of fractions the number of goals met was the same. The online adaptive recontouring and replanning process was carried out in 19 min on average. Significant improvements in dosimetry are possible with the Ethos online adaptive system in prostate radiotherapy.

Performance Testing and Rapid Solidification Behavior of Stainless Steel Powders Prepared by Gas Atomization.

Gas atomization is a widely used method to produce the raw powder materials for additive manufacturing (AM) usage. After the metal alloy is melted to fusion, gas atomization involves two relatively independent processes: liquid breakup and droplet solidification. In this paper, the solidification behavior of powder during solidification is analyzed by testing the powder's properties and observing microstructure of a martensitic stainless steel (FeCrNiBSiNb). The powder prepared by gas atomization has high sphericity and smooth surface, and the yield of qualified fine powder is 35%. The powder has typical rapid solidification structure. Collision between powders not only promotes nucleation, but also produces more satellite powder. The segregation of elements in powder is smaller as the result of high cooling rate which can reaches 4.2 × 105 K/s in average. Overall, the powder prepared by gas atomization is found to have good comprehensive properties, desired microstructure, and accurate chemical component, and it is suitable for various additive manufacturing techniques.

Quality assessment of Succus Bambusae oral liquids based on gas chromatography/mass spectrometry fingerprints and chemometrics.

RATIONALE: Succus Bambusae is consumed as a kind of herbal medicine and natural beverage in China. However, the current quality standards for Succus Bambusae are low and lack safety indicators, which makes it difficult to effectively guarantee its quality. Therefore, it is of great significance to study the identification and quality control technology for the product. METHODS: We have developed a set of qualitative and quantitative methods based on gas chromatography/mass spectrometry (GC/MS) for the analysis of volatile components in Succus Bambusae oral liquid (SBOL). Combining GC/MS fingerprint analysis and related chemometrics algorithms, with similarity evaluation, Hotelling T2 and distance to Model X (DModX) as criteria, the quality consistency of different batches was evaluated, and SBOL samples from different manufacturers were differentiated. RESULTS: Twenty-nine volatile components were preliminarily identified from 40 batches of SBOL samples from six manufacturers, and six Q-markers (Quality Markers) for the SBOLs were discussed and determined using GC/MS. The products from different manufacturers were distinguished using chemometrics. CONCLUSIONS: The results showed that the quality of the SBOL samples from different batches and different manufacturers fluctuated greatly, which suggested that research into the raw materials and manufacturing techniques should be strengthened to improve the quality of SBOL and ensure its quality consistency.

Design, synthesis and biological evaluation of indole-based [1,2,4]triazolo[4,3-a] pyridine derivatives as novel microtubule polymerization inhibitors.

A series of indole-based [1,2,4]triazolo [4,3-a]pyridine derivatives was designed and synthesized as novel microtubulin polymerization inhibitors by using a conformational restriction strategy. These compounds exhibited moderate to potent anti-proliferative activities against a panel of cancer cell lines (HeLa, A549, MCF-7 and HCT116). Among them, compound 12d featuring a N-methyl-5-indolyl substituent at the C-6 position of the [1,2,4]triazolo [4,3-a]pyridine core exhibited the highest antiproliferative activity with the IC50 values ranging from 15 to 69 nM, and remarkable inhibitory effect on tubulin polymerization with an IC50 value of 1.64 µM. Mechanistic studies revealed that compound 12d induced cellular apoptosis and cell cycle arrest at the G2/M phase in a dose-dependent fashion. Moreover, compound 12d significantly suppressed wound closure and disturbed microtubule networks.

Realizing Fast Charge Diffusion in Oriented Iron Carbodiimide Structure for High-Rate Sodium-Ion Storage Performance.

Iron carbodiimide (FeNCN) belongs to a type of metal compounds with a more covalent bonding structure compared to common transition metal oxides. It could provide possibilities for various structural designs with improved charge-transfer kinetics in battery systems. Moreover, these possibilities are still highly expected for promoting enhancement in rate performance of sodium (Na)-ion battery. Herein, oriented FeNCN crystallites were grown on the carbon-based substrate with exposed {010} faces along the [001] direction (O-FeNCN/S). It provides a high Na-ion storage capacity with excellent rate capability (680 mAh g-1 at 0.2 A g-1 and 360 mAh g-1 at 20 A g-1), presenting rapid charge-transfer kinetics with high contribution of pseudocapacitance during a typical conversion reaction. This high rate performance is attributed to the oriented morphology of FeNCN crystallites. Its orientation along [001] maintains preferred Na-ion diffusion along the two directions in the entire morphology of O-FeNCN/S, supporting fast Na-ion storage kinetics during the charge/discharge process. This study could provide ideas toward the understanding of the rational structural design of metal carbodiimides for attaining high electrochemical performance in future.

Comparative chloroplast genomes: insights into the evolution of the chloroplast genome of Camellia sinensis and the phylogeny of Camellia.

BACKGROUND: Chloroplast genome resources can provide useful information for the evolution of plant species. Tea plant (Camellia sinensis) is among the most economically valuable member of Camellia. Here, we determined the chloroplast genome of the first natural triploid Chinary type tea ('Wuyi narcissus' cultivar of Camellia sinensis var. sinensis, CWN) and conducted the genome comparison with the diploid Chinary type tea (Camellia sinensis var. sinensis, CSS) and two types of diploid Assamica type teas (Camellia sinensis var. assamica: Chinese Assamica type tea, CSA and Indian Assamica type tea, CIA). Further, the evolutionary mechanism of the chloroplast genome of Camellia sinensis and the relationships of Camellia species based on chloroplast genome were discussed. RESULTS: Comparative analysis showed the evolutionary dynamics of chloroplast genome of Camellia sinensis were the repeats and insertion-deletions (indels), and distribution of the repeats, indels and substitutions were significantly correlated. Chinese tea and Indian tea had significant differences in the structural characteristic and the codon usage of the chloroplast genome. Analysis of sequence characterized amplified region (SCAR) using sequences of the intergenic spacers (trnE/trnT) showed none of 292 different Camellia sinensis cultivars had similar sequence characteristic to triploid CWN, but the other four Camellia species did. Estimations of the divergence time showed that CIA diverged from the common ancestor of two Assamica type teas about 6.2 Mya (CI: 4.4-8.1 Mya). CSS and CSA diverged to each other about 0.8 Mya (CI: 0.4-1.5 Mya). Moreover, phylogenetic clustering was not exactly consistent with the current taxonomy of Camellia. CONCLUSIONS: The repeat-induced and indel-induced mutations were two important dynamics contributed to the diversification of the chloroplast genome in Camellia sinensis, which were not mutually exclusive. Chinese tea and Indian tea might have undergone different selection pressures. Chloroplast transfer occurred during the polyploid evolution in Camellia sinensis. In addition, our results supported the three different domestication origins of Chinary type tea, Chinese Assamica type tea and Indian Assamica type tea. And, the current classification of some Camellia species might need to be further discussed.