Differential pathogenic and molecular features in neurological infection of SARS-CoV-2 Omicron BA.5.2 and BA.2.75 and Delta.

The Coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remains a global threat, exacerbated by the emergence of viral variants. Two variants of SARS-CoV-2, Omicron BA.2.75 and BA.5, led to global infection peaks between May 2022 and May 2023, yet their precise characteristics in pathogenesis are not well understood. In this study, we compared these two Omicron sublineages with the previously dominant Delta variant using a human angiotensin-converting enzyme 2 knock-in mouse model. As expected, Delta exhibited higher viral replication in the lung and brain than both Omicron sublineages which induced less severe lung damage and immune activation. In contrast, the Omicron variants especially BA.5.2 showed a propensity for cellular proliferation and developmental pathways. Both Delta and BA.5.2 variants, but not BA.2.75, led to decreased pulmonary lymphocytes, indicating differential adaptive immune response. Neuroinvasiveness was shared with all strains, accompanied by vascular abnormalities, synaptic injury, and loss of astrocytes. However, Immunostaining assays and transcriptomic analysis showed that BA.5.2 displayed stronger immune suppression and neurodegeneration, while BA.2.75 exhibited more similar characteristics to Delta in the cortex. Such differentially infectious features could be partially attributed to the weakened interaction between Omicron Spike protein and host proteomes decoded via co-immunoprecipitation followed by mass spectrometry in neuronal cells. Our present study supports attenuated replication and pathogenicity of Omicron variants but also highlights their newly infectious characteristics in the lung and brain, especially with BA.5.2 demonstrating enhanced immune evasion and neural damage that could exacerbate neurological sequelae.

Prevalence and progression of pneumonia in immunocompetent adults with varicella.

Pneumonia is the most common complication of varicella infections. Although previous studies have tended to focus mainly on immunocompromised patients, varicella pneumonia can also occur in healthy adults. Therefore, in this study, we aimed to assess the progression of varicella pneumonia in immunocompetent hosts. This retrospective study involved immunocompetent adult outpatients with varicella who attended the adult Fever Emergency facility of Peking University Third Hospital from April 1, 2020, to October 31, 2022. Varicella pneumonia was defined as a classic chickenpox-type rash in patients with infiltrates on chest computed tomography. The study included 186 patients, 57 of whom had a contact history of chickenpox exposure. Antiviral pneumonia therapy was administered to 175 patients by treating physicians. Computed tomography identified pneumonia in 132 patients, although no deaths from respiratory failure occurred. Seventy of the discharged patients were subsequently contacted, all of whom reported being well. Follow-up information, including computed tomography findings, was available for 37 patients with pneumonia, among whom 24 reported complete resolution whereas the remaining 13 developed persistent calcifications. Notably, we established that the true incidence of varicella pneumonia is higher than that previously reported, although the prognosis for immunocompetent hosts is generally good.

Mining biomarkers from routine laboratory tests in clinical records associated with air pollution health risk assessment.

Clinical laboratory in hospital can produce amounts of health data every day. The purpose of this study was to mine biomarkers from clinical laboratory big data associated with the air pollution health risk assessment using clinical records. 13, 045, 629 clinical records of all 27 routine laboratory tests in Changsha Central Hospital, including ALB, TBIL, ALT, DBIL, AST, TP, UREA, UA, CREA, GLU, CK, CKMB, LDL-C, TG, TC, HDL-C, CRP, WBC, Na, K, Ca, Cl, APTT, PT, FIB, TT, RBC and those daily air pollutants concentration monitoring data of Changsha, including PM2.5, PM10, SO2, NO2, CO, and O3 from 2014 to 2016, were retrieved. The moving average method was used to the biological reference interval was established. The tests results were converted into daily abnormal rate. After data cleaning, GAM statistical model construction and data analysis, a concentration-response relationship between air pollutants and daily abnormal rate of routine laboratory tests was observed. Our study found that PM2.5 had a stable association with TP (lag07), ALB (lag07), ALT (lag07), AST (lag07), TBIL (lag07), DBIL (lag07), UREA (lag07), CREA (lag07), UA (lag07), CK (lag 06), GLU (lag07), WBC (lag07), Cl (lag07) and Ca (lag07), (P < 0.05); O3 had a stable association with AST (lag01), CKMB (lag06), TG (lag07), TC (lag05), HDL-C (lag07), K (lag05) and RBC (lag07) (P < 0.05); CO had a stable association with UREA (lag07), Na (lag7) and PT (lag07) (P < 0.05); SO2 had a stable association with TP (lag07) and LDL-C (lag0) (P < 0.05); NO2 had a stable association with APTT (lag7) (P < 0.05). These results showed that different air pollutants affected different routine laboratory tests and presented different pedigrees. Therefore, biomarkers mined from routine laboratory tests may potentially be used to low-cost assess the health risks associated with air pollutants.

Efficient Near-Infrared Luminescence in Lanthanide-Doped Vacancy-Ordered Double Perovskite Cs2 ZrCl6 Phosphors via Te4+ Sensitization.

All-inorganic lead-free perovskite-derivative metal halides have shown great promise in optoelectronics, however, it remains challenging to realize efficient near-infrared (NIR) luminescence in these materials. Herein, we report a novel strategy based on Te4+ /Ln3+ (Ln=Er, Nd, and Yb) co-doping to achieve efficient NIR luminescence in vacancy-ordered double perovskite Cs2 ZrCl6 phosphors, which are excitable by a low-cost near-ultraviolet light-emitting diode (LED) chip. Through sensitization by the spin-orbital allowed 1 S0 →3 P1 transition of Te4+ , intense and multi-wavelength NIR luminescence originating from the 4f→4f transitions of Er3+ , Nd3+ , and Yb3+ was acquired, with a quantum yield of 6.1 % for the Er3+ emission. These findings provide a general approach to achieve efficient NIR emission in lead-free metal halides through ns2 -metal and lanthanide ion co-doping, thereby opening up a new avenue for exploring NIR-emitting perovskite derivatives towards versatile applications such as NIR-LEDs and bioimaging.

[The Effects of Different Adaptive Statistical Iterative Reconstruction-V and Convolution Kernel Parameters on Auto-Segmentation Stability in CT Images].

Objective The study aims to investigate the effects of different adaptive statistical iterative reconstruction-V（ ASiR-V） and convolution kernel parameters on stability of CT auto-segmentation which is based on deep learning. Method Twenty patients who have received pelvic radiotherapy were selected and different reconstruction parameters were used to establish CT images dataset. Then structures including three soft tissue organs (bladder, bowelbag, small intestine) and five bone organs (left and right femoral head, left and right femur, pelvic) were segmented automatically by deep learning neural network. Performance was evaluated by dice similarity coefficient（ DSC） and Hausdorff distance, using filter back projection(FBP) as the reference. Results Auto-segmentation of deep learning is greatly affected by ASIR-V, but less affected by convolution kernel, especially in soft tissues. Conclusion The stability of auto-segmentation is affected by parameter selection of reconstruction algorithm. In practical application, it is necessary to find a balance between image quality and segmentation quality, or improve segmentation network to enhance the stability of auto-segmentation.

Effective method to reduce the normal brain dose in single-isocenter hypofractionated stereotactic radiotherapy for multiple brain metastases.

BACKGROUND AND PURPOSE: Island blocking and dose leakage problems will lead to unnecessary irradiation to normal brain tissue (NBT) in hypofractionated stereotactic radiotherapy (HSRT) for multiple brain metastases (BM) with single-isocenter volumetric modulated arc therapy (VMAT). The present study aimed at investigating whether reducing the number of metastases irradiated by each arc beam could minimize these two problems. MATERIALS AND METHODS: A total of 32 non-small-cell lung cancer (NSCLC) patients with multiple BM received HSRT (24-36 Gy/3 fractions) with single-isocenter VMAT, where each arc beam only irradiated partial metastases (pm-VMAT), were enrolled in this retrospective study. Conventional single-isocenter VMAT plans, where each arc beam irradiated whole metastases (wm-VMAT), was regenerated and compared with pm-VMAT plans. Furthermore, the clinical efficacy and toxicities were evaluated. RESULTS: Pm-VMAT achieved similar target coverage as that with wm-VMAT, with better dose fall-off (P < 0.001) and NBT sparing (P < 0.001). However, pm-VMAT resulted in more monitor units (MU) and longer beam-on time (P < 0.001). The intracranial objective response rate and disease control rate for all patients were 75% and 100%, respectively. The local control rates at 1 year and 2 year were 96.2% and 60.2%, respectively. The median progression-free survival and overall survival were 10.3 months (95% confidence interval [CI] 6.8-13.2) and 18.5 months (95% CI 15.9-20.1), respectively. All treatment-related adverse events were grade 1 or 2, and 3 lesions (2.31%) from 2 patients (6.25%) demonstrated radiation necrosis after HSRT. CONCLUSION: HSRT with pm-VMAT is effective and has limited toxicities for NSCLC patients with multiple BM. Pm-VMAT could provide better NBT sparing while maintaining target dose coverage.

Multi-Dopant Engineering in Perovskite Cs2SnCl6: White Light Emitter and Spatially Luminescent Heterostructure.

Bi3+/Te4+ co-doped Cs2SnCl6 with dual emission spectrum (i.e., 450 and 575 nm) was achieved by a modified solution method, which can overcome the phase separation in the previous method for Cs2SnCl6 crystal growth. The two emission peaks arising from the two dopants Bi3+ and Te4+ have distinct photoluminescence (PL) lifetimes. Thus, the control of dopant ratio or PL delay time will regulate the PL intensity ratio between 450 and 575 nm peaks leading to adjustable emission color. The energy transfer between the two emission centers, which is confirmed by the optical spectra and PL lifetime, has a critical distance around 7.8 nm with a maximum of 50% transfer efficiency. The Bi3+/Te4+ co-doped Cs2SnCl6 with superior stability in water and aqua regia was fabricated into a single-phase white light-emitting diode. In the meantime, various luminescent heterostructures were obtained by epitaxial Cs2SnCl6 crystal growth with different dopants, which can broaden the study of composition engineering in halide perovskites.

Tumour necrosis factor-α and myoglobin associated with the recovery time of coronary artery lesions in Kawasaki disease patients.

AIM: To assess the relationship between clinical parameters and medium term recovery time of coronary artery lesions (CALs). METHODS: In total, 344 Kawasaki disease patients were screened and 311 Kawasaki disease patients were included and followed-up for the next 2 years. Clinical records, clinical parameters and inflammatory biomarkers were collected for all subjects. RESULTS: Tumour necrosis factor (TNF)-α and myoglobin (MYO) levels in patients without recovery from CALs were significantly higher than those without CALs and with recovery from CALs. Kaplan-Meier survival analysis showed that in the high-TNF-α group, the estimated median time to recovery (5.0 months, 95% confidence interval (CI) 1.436-8.564) is significantly longer than the low-TNF-α group (2.00 months, 95% CI: 0.633-3.367, P = 0.044). Also, the estimated median time (5.0 months, 95% CI: 1.836-8.164) in the high-MYO group is significantly longer than the low-MYO group (2.00 months, 95% CI: 0.405-3.595, P = 0.002). Cox regression analysis showed independent factors for recovery of CALs included age, left coronary artery to aortic annulus ratio, TNF-α and MYO levels. CONCLUSIONS: These findings suggest that clinical parameters such as age, left coronary artery to aortic annulus ratio, TNF-α and MYO levels associate with medium term recovery time of CALs and could help in the design of a clinical strategy for the surveillance and prevention of late cardiovascular events.

Moisture-Resistant Mn4+ -Doped Core-Shell-Structured Fluoride Red Phosphor Exhibiting High Luminous Efficacy for Warm White Light-Emitting Diodes.

K2 TiF6 :Mn4+ is a highly efficient narrow-band emission red phosphor with promising applications in white light-emitting diodes (LEDs) and wide-gamut displays. Nevertheless, the poor moisture-resistant properties of this material hinder commercialization. A convenient reverse cation-exchange strategy is introduced for constructing a core-shell-structured K2 TiF6 :Mn4+ @K2 TiF6 phosphor. The outer K2 TiF6 shell acts as a shield for preventing moisture in the air from hydrolyzing the internal MnF6 2- group, while effectively cutting off the path of energy migration to surface defects, thereby increasing the emission efficiency (especially for the phosphors doped with high concentrations of Mn4+ ). Employed as a red phosphor, the packaged white LED exhibits an extraordinarily high luminous efficacy of 162 lm W-1 , a correlated color temperature (CCT) of 3510 K, and a color rendering index of 93 (Ra ). Aging tests performed on this device at 85 °C and 85 % humidity for 480 h retain up to 89 % luminous efficacy. The findings could facilitate commercial application of K2 TiF6 :Mn4+ @K2 TiF6 phosphor.

Temperature dependent carrier localization in AlGaInN near-ultraviolet light-emitting diodes.

We investigate the carrier localization effect in the low-indium ultraviolet AlGaInN light-emitting diodes with a 365 nm peak and a wide yellow luminous band over the visible range. Temperature-dependent electroluminescence spectra (EL) are measured under a wide range of temperature. We found that carrier localization effect relies on the carrier mobility and manifests itself by altering several macroscopic quantities, such as ELs and electrical resistance of the device. Under moderate injection densities, plots of EL peak energy vs. temperatures exhibits S-shapes. At low temperatures, line-width broadening in EL spectra and irregular humps in I-V curves were observed at similar level of injection densities. Both phenomena diminish as temperature increases and eventually disappear at room temperature. All the results stem from carrier localization and following delocalization effect. It suggests that the carrier mobility determine the degree of carrier localization effect - inactive carriers tend to be localized at low temperature but escape at high temperature from bindings of localization centers. As a result, carrier localization is intense only at low temperature for low-indium devices.

Dual-Emission Luminescence of Magnesium Coordination Polymers Based on Mixed Organic Ligands.

Presented herein are two luminescent magnesium coordination polymers (Mg-CPs), namely [Mg2 (H2O)2 (2-NDC)4 (1,10-phen)2] (1) and [Mg2 (H2O)(1,4-NDC)2 (1,10-phen)] (2), in which 2-NDCH=2-naphthalenecarboxylic acid, 1,4-NDCH2 =1,4-naphthalene dicarboxylic acid, and 1,10-phen=1,10-phenanthroline. Based on the mixed ligands, the title compounds exhibit linker-based photoluminescence (PL) properties thanks to the unique configuration of the Mg(2+) ions. The two compounds show interesting dual emission on excitation of the different luminophores of the mixed linkers. In particular, the emissions of compound 2 could be tuned from green to yellow simply by varying the excitation energies. Furthermore, 2 could be excited by using a commercial λ=450 nm blue LED chip to generate white-light emission, which allows the fabrication of a white-light-emitting diode (WLED) with 20 lm W(-1) luminous efficacy. This work may provide a new method for designing tunable PL CPs by using the low-cost and abundant magnesium ion.

Optical enhancement brought by doping Gd(3+) ions into Ce: YAG ceramics for indoor white light-emitting diodes.

We dope Gd(3+) ions into Ce: YAG ceramics to induce red-shift in the photoluminescence, of which the degree is proportional to the Gd(3+) doping concentration. This kind of ceramic, when collaborating with InGaN blue chips, proves to be a promising fluorescent material of white light-emitting diodes, for not only its high in-line transmittance and decent quantum yield, but also the improvement in color rendering ability brought by the red-shift, which enhances the ratio of red portion in spectra. We demonstrate that 10% is the optimized value of Gd(3+) doping concentration, as it provides the maximum color rendering index of ~78 with luminous efficiency as high as 128 lm/W.

Defects dynamics during ageing cycles of InGaN blue light-emitting diodes revealed by evolution of external quantum efficiency--current dependence.

We report in detail the defect dynamics in the active region by monitoring the external quantum efficiency (EQE) - injection current curves, I-V curves, and electroluminescence spectra during the ageing test, under a forward current of 850 mA (85 A/cm2), room temperature. We apply a two-level model to analyze the EQE curves and the electroluminescence spectra. The results suggest that high injection density during the ageing may reduce the density of the Shockley-Reed-Hall nonradiative recombination centers and enhance the carrier mobility and diffusion length. The former effect would directly lead to initial surge of EQE, whereas the latter would enhance the effect of extended defects which leads to reduction in peak EQE and increase in EQE droop rate.

Immunogenicity of recombinant human bocavirus-1,2 VP2 gene virus-like particles in mice.

Human bocavirus (HBoV), a recently identified pathogen with a worldwide distribution is closely related to paediatric acute respiratory infection and gastroenteritis. The present study was performed to evaluate the immunogenicity of HBoV1 and HBoV2 virus-like particles (VLPs) as vaccine candidates in mice. Both HBoV1 and HBoV2 VLPs were expressed in the bacmid virus­SF9 cell system. Mice were inoculated three times at 3-week intervals with HBoV VLPs at one dose intramuscular (i.m.) or intradermal (i.d.) with or without the addition of the alum adjuvant. ELISA was used to detected antibody, and ELISPOT was used to test cellular immune responses. HBoV-specific IgG antibodies were induced and alum adjuvant improved the antibody titres and avidity, while the inoculation pathway had no influence. T helper type 1/ type 2 immune responses were balanced induced by HBoV1 VLPs but not HBoV2 VLPs. Serum IgG antibody cross-reactivity rates of the two subtypes were similar, but cross-reactions of HBoV1 immunization groups were higher. The single i.m. group had more interferon-γ-secreting splenocytes. These data indicate that HBoV VP2 VLPs have good immunogenicity with induction of strong humoral and cellular immune responses, and they may be potential candidate vaccines for HBoV infection.

Introduction on the fabrication technique of phosphor in glass by tape-casting and investigation on the chromaticity property.

We introduce a new fabrication technique of phosphor in glass (PiG) for light-emitting diodes (LEDs) employing the tape-casting. Through the detailed process described herein and the measurement results, it is clear that the PiG-on-glass not only share the same characteristic of those obtained from other techniques or the bulk PiG, but with more precisely controlled width from a few to hundreds micrometers. The samples are mounted on blue InGaN LED chips to test the color properties of the white light. Besides, we established an empirical model that could predict the final color properties of LEDs solely by the phosphor concentration of phosphor glass under certain conditions. This model would greatly facilitate the design of PiG-based LEDs.

Study on the correlations between color rendering indices and the spectral power distribution.

The intrinsic spectrally resolved sensitivity (ISRS) of color rendering indices (CRIs) is investigated by using spectral loss simulations. It is demonstrated that R(a) exhibits large sensitivities around 444, 480, 564, and 622 nm, while for R(9) the sensitivity peaks are around 461, 581 and 630 nm, which all shift slightly with the correlated color temperature. If considering the ISRS as a bridge between the spectral power distribution of LED and its CRI, one could obtain a high CRI by minimizing the deviation between the shapes of the illuminant spectrum and the reference spectrum, both after modulations by the ISRS as a weighting function. This approach, recommended as a guideline for the spectra design aiming at a high CRI, is described and justified in depth via a mathematical model. This method is spectra-oriented and could largely facilitate the spectra design.

Long-term exposure to PM2.5 leads to mitochondrial damage and differential expression of associated circRNA in rat hepatocytes.

Fine particulate matter (PM2.5) is one of the four major causes of mortality globally. The objective of this study was to investigate the mechanism underlying liver injury following exposure to PM2.5 and the involvement of circRNA in its regulation. A PM2.5 respiratory tract exposure model was established in SPF SD male rats with a dose of 20 mg/kg, and liver tissue of rats in control group and PM2.5-exposed groups rats were detected. The results of ICP-MS showed that Mn, Cu and Ni were enriched in the liver. HE staining showed significant pathological changes in liver tissues of PM2.5-exposed group, transmission electron microscopy showed significant changes in mitochondrial structure of liver cells, and further mitochondrial function detection showed that the PM2.5 exposure resulted in an increase in cell reactive oxygen species content and a decrease in mitochondrial transmembrane potential, while the expression of SOD1 and HO-1 antioxidant oxidase genes was upregulated. Through high-throughput sequencing of circRNAs, we observed a significant down-regulation of 10 and an up-regulation of 17 circRNAs in the PM2.5-exposed groups. The functional enrichment and pathway analyses indicated that the differentially expressed circRNAs by PM2.5 exposure were primarily associated with processes related to protein ubiquitination, zinc ion binding, peroxisome function, and mitochondrial regulation. These findings suggest that the mechanism underlying liver injury induced by PM2.5-exposure may be associated with mitochondrial impairment resulting from the presence of heavy metal constituents. Therefore, this study provides a novel theoretical foundation for investigating the molecular mechanisms underlying liver injury induced by PM2.5 exposure.

Nomogram for predicting coronary artery lesions in patients with Kawasaki disease.

BACKGROUND: Coronary artery lesions are the most important complications of Kawasaki disease. Approximately 25-30% of untreated patients develop coronary artery disease, which can lead to long-term cardiovascular sequelae. AIM: The aim of this study is to evaluate the risk factors for coronary artery lesions in Kawasaki disease and to construct a nomogram for predicting the likelihood of developing such lesions. METHODS: Data from 599 patients between January 2012 and June 2020 were reviewed retrospectively. Patients were randomly assigned to the training set (n = 450) and the validation set (n = 149). A comparison of clinical features and laboratory data was performed, followed by multivariate logistic regression analysis to identify independent risk factors and develop the nomogram. The predictive efficiency of the nomogram was evaluated using the calibration curve, area under the receiver operating characteristic curve (AUC), C-index, and decision curve analysis (DCA). RESULTS: Intravenous immunoglobulin (IVIG) resistance, delayed IVIG treatment, C-reactive protein, and neutrophil/lymphocyte ratio were identified as independent risk factors for the development of coronary artery lesions. The nomogram was constructed based on these four variables. The calibration curve of the nomogram showed a high degree of agreement between the predicted probability and the actual probability. The AUC of the nomogram in the training and validation set was 0.790 and 0.711, respectively. In addition, DCA revealed that the nomogram provided a significant net benefit, further supporting its clinical utility. CONCLUSIONS: The constructed nomogram demonstrates a strong and reliable performance in predicting coronary artery lesions, which enables clinicians to make timely and tailored clinical decisions.

Boosting the Self-Trapped Exciton Emission in Alloyed Cs2 (Ag/Na)InCl6 Double Perovskite via Cu+ Doping.

Fundamental understanding of the effect of doping on the optical properties of 3D double perovskites (DPs) especially the dynamics of self-trapped excitons (STEs) is of vital importance for their optoelectronic applications. Herein, a unique strategy via Cu+ doping to achieve efficient STE emission in the alloyed lead-free Cs2 (Ag/Na)InCl6 DPs is reported. A small amount (1.0 mol%) of Cu+ doping results in boosted STE emission in the crystals, with photoluminescence (PL) quantum yield increasing from 19.0% to 62.6% and excitation band shifting from 310 to 365 nm. Temperature-dependent PL and femtosecond transient absorption spectroscopies reveal that the remarkable PL enhancement originates from the increased radiative recombination rate and density of STEs, as a result of symmetry breakdown of the STE wavefunction at the octahedral Ag+ site. These findings provide deep insights into the STE dynamics in Cu+ -doped Cs2 (Ag/Na)InCl6 , thereby laying a foundation for the future design of new lead-free DPs with efficient STE emission.

Analysis of the amplitude changes and baseline shifts of respiratory motion using intra-fractional CBCT in liver stereotactic body radiation therapy.

PURPOSE: Using intra-fractional cone-beam CT (CBCT) to evaluate the amplitude changes and baseline shifts of respiratory motion in liver stereotactic body radiation therapy (SBRT). METHODS: The amplitude changes and baseline shifts of respiratory motion for 24 liver patients were evaluated by the four-dimensional (4D) CT, inter- and intra-fractional CBCT. The difference of the average liver position errors among all treatment fractions and the 4D CT representthe baseline shifts. According to the baseline shifts, the ITV to PTV margin was recalculated and the plan was re-designed to compare the dosimetric variation. RESULTS: The systematic and random errors of the baseline shifts for intra-fractional CBCT in the left-right (LR), superior-inferior (SI), and anterior-posterior (AP) directions were 0.99/1.60 mm, 2.03/2.46 mm, and 1.02/2.07 mm, respectively. The new ITV to PTV margins should be 4.0 mm, 7.0 mm, and 4.0 mm, respectively. The amplitude change of motion between the 4D CT and the intra-fractional CBCT was 1.03 ± 4.35 mm, with 31% of fractions exceeding 5 mm. To achieve the same dose coverage of the new PTV, the Dmean, V50, V40, V30, V25 of normal liver and maximum dose of the duodenum were significantly different. CONCLUSIONS: Significant amplitude changes and baseline shifts of motion occurred during dose delivery compared with those in 4D CT. Using the ITV to PTV margin of 4.0 mm (LR), 7.0 mm (SI), and 4.0 mm (AP) can ensure the target dose coverage and keep the dose constrain of normal tissues at an acceptable level.