ObjectiveThis paper aims to investigate the role of NDC80 kinetochore complex component （NUF2） and magnesium homeostasis in ovarian cancer cell apoptosis， as well as the regulatory mechanism of gypenoside L （Gyp-L） on NUF2 and magnesium homeostasis. MethodsOvarian cancer OVCAR3 cells were divided into a blank control group， a low-concentration Gyp-L group （50 µmol·L^-1）， a high-concentration Gyp-L group （100 µmol·L^-1）， and a cisplatin （15 µmol·L^-1） group. The migration， proliferation， and apoptosis capabilities of OVCAR3 cells were evaluated through cell scratch assays， clonal experiments， and terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay （TUNEL） staining. Differentially expressed genes of ovarian cancer were screened by using the Gene Expression Omnibus （GEO） database. The interaction relationships of differentially expressed genes and proteins were analyzed via the Search Tool for Recurring Instances of Neighbouring Genes （STRING） database. The prognostic survival analysis was performed by using the Tumor Immune Estimation Resource （TIMER） database， and the differential expression levels of genes were validated with the Gene Expression Profiling Interactive Analysis （GEPIA） database. The mRNA expression levels of NUF2， magnesium homeostasis-related indicators， such as magnesium transporter 1 （MAGT1）， non-imprinted in Prader-Willi/Angelman syndrome 1 （NIPA1）， NIPA-like domain containing 1 （NIPAL1）， as well as apoptosis-related indicators B cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax） in OVCAR3 cells， were detected by real-time quantitative polymerase chain reaction （Real-time PCR）. The protein expression levels of NUF2， MAGT1， NIPA1， NIPAL1， Bcl-2， and Bax in OVCAR3 cells were quantitatively analyzed by ProteinSimple WES. A model of overexpression of NUF2 was constructed， and Gyp-L intervention was performed. The molecular mechanism by which Gyp-L induces ovarian cancer cell apoptosis by regulating NUF2 and influencing magnesium homeostasis was quantitatively analyzed and detected through cell cloning， TUNEL staining， Real-time PCR， and ProteinSimple WES. Finally， the Mg²⁺ content and protein synthesis efficiency were detected by immunofluorescence. ResultsGyp-L significantly inhibited the migration and proliferation capabilities of OVCAR3 cells and promoted their apoptosis （P<0.05）. Overexpression of NUF2 markedly increased the expression levels of MAGT1， NIPA1， NIPAL1， and Bcl-2， while reducing the expression level of Bax （P<0.05）. It also significantly elevated intracellular Mg²⁺ content and protein synthesis efficiency and simultaneously inhibited apoptosis （P<0.05）. Gyp-L could reverse the magnesium homeostasis imbalance and apoptosis inhibition caused by the overexpression of NUF2， downregulating the expression levels of NUF2， MAGT1， NIPA1， NIPAL1， and Bcl-2 （P<0.05）， while upregulating the expression level of Bax （P<0.05）. ConclusionGyp-L can inhibit the occurrence of ovarian cancer， and its mechanism may involve inhibiting the expression of NUF2 to maintain magnesium homeostasis and inducing apoptosis of ovarian cancer cells.

ObjectiveThis study aims to systematically analyze the blood-absorbed components and metabolic profiles of Xiebaisan（XBS） in normal and chronic bronchitis （CB） mice using ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS）， while comparing differences between the two states. MethodsThirty female BABL/c mice were randomly divided into the normal group， the normal drug administration group， the CB group， the CB drug administration group and the dexamethasone group， with 6 mice in each group. The CB mouse model was established by inducing with ovalbumin （OVA）. The mice in the normal drug administration group and the CB drug administration group started to be gavaged with XBS（13.2 g·kg^-1） from the 21^st day， and the dexamethasone group mice were simultaneously gavaged with dexamethasone （0.5 mg·kg^-1） until the end of the 35^th day of the experiment. Subsequently， serum samples were collected and evaluated for their efficacy， based on the pharmacological evaluation indicators， to determine the efficacy of XBS in treating CB. Then the UPLC-Q-Exactive Orbitrap MS was employed to identify and analyze the chemical constituents， blood-absorbed components， and metabolites of XBS. Chemometric analysis was conducted to reveal metabolic profile differences under "dual states". Concurrently， Real-time PCR technology was utilized to detect the expression levels of key liver metabolic enzymes CYP2E1， CYP3A1， UGT1A1， and UGT1A6. ResultsA total of 28 prototype components and 158 metabolites （including 48 phase Ⅰ metabolites and 110 phase Ⅱ metabolites） of XBS were unambiguously identified in the serum of normal mice. Additionally， a comprehensive characterization was performed on a total of 32 prototype components and 178 metabolites （including 50 phase Ⅰ metabolites and 128 phase Ⅱ metabolites） of XBS in the serum of CB mice. Among them， 27 prototype components were detected in both states， including 12 flavonoids， 2 alkaloids， 3 triterpenes， 4 organic acids， 3 amides， 1 stilbene and 2 other compounds. The chemometrics analysis revealed no significant difference in the prototype components and metabolites of XBS between normal and CB mice； however， there was a significant increase in the in-vivo exposure of XBS in CB mice. Compared to normal mice， the levels of phase Ⅰ metabolites such as oxidation， reduction and methylation of blood components of XBS as well as phase Ⅱ metabolites of glucuronidation showed significant changes in CB mice. Real-time PCR further confirmed that these alterations were attributed to the upregulation of CYP2E1 （P<0.05）， CYP3A1 （P>0.05）， UGT1A1 （P<0.01） and UGT1A6 （P<0.01） enzymes expression in the liver of CB mice. ConclusionThis study elucidated the disparities in the levels of the blood-absorbed components and metabolic profiles of XBS in normal and CB mice， especially in oxidation， reduction， methylation in phase Ⅰ metabolism and glucoaldehyde acidification in phase Ⅱ metabolism. And there are related to the differences in the expression levels of phase Ⅰ and phase Ⅱ metabolic enzymes CYP2E1， CYP3A1， UGT1A1 and UGT1A6 in the liver.

Objective To explore the characteristics and temporal trends of renal cancer burden among Chinese adolescents and young adults(AYA), and predict its future epidemic situation, so as to provide evidence for formulating public health policies and reducing the disease burden.Methods Epidemiological data of renal cancer in Chinese AYA(15-39 years)from 1990 to 2021 were obtained from the Global Burden of Disease(GBD)2021 database. The current status and trends of renal cancer in this population were analyzed. The incidence and mortality trends of renal cancer in AYA over the next 15 years was predicted with ARIMA model. The impacts of smoking and body mass index(BMI)on the disease burden were evaluated. Results In 2021, the incidence(1.12/100000), prevalence(8.61/100000), mortality(0.19/100000), and disability-adjusted life years(DALYs)(11.37/100000 person-years)of renal cancer in Chinese AYA were significantly higher than those in 1990 and above the global average. All burden indicators and their increases were higher in males than in females, and the burden accelerated after 30 years of age. The incidence rose steadily from 2003 to 2021(APC=3.95), while the mortality increased significantly from 2013 to 2021(APC=2.40). The burden attributed to high BMI continued to rise, and the smoking-attributable fraction in males first increased then decreased. ARIMA predicted that the incidence of renal cancer in Chinese AYA males would accelerate, while the mortality in both genders would stabilize. Conclusion The burden of renal cancer in Chinese AYA shows a significant upward trend. Males and people over 30 years old are key targets for prevention and control. Strengthening the management of risk factors(smoking, high BMI)and promoting early diagnosis and treatment are essential to reduce the disease burden.

Objective To screen the aging-related genes associated with prognosis in clear cell renal cell carcinoma (ccRCC), construct a prognostic model to optimize risk stratification, explore the underlying mechanisms, and provide reference for individualized diagnosis and treatment of this disease.Methods RNA sequencing data, clinical data, and aging-related genes of ccRCC were obtained from databases.After differentially expressed genes were screened, a prognostic model was constructed using Cox regression.The performance of the model was evaluated with Kaplan-Meier curves and receiver operating characteristic (ROC) curves, and validated with clinical features.Key genes were verified with immunohistochemistry.Results A total of 70 differentially expressed genes were identified, including IFI16, PECAM1, and ABCB1 as key genes, and a model formula was constructed.Internal and external validations showed that the survival rate of the high-risk group was significantly lower, with the area under the ROC curve (AUC)＞0.6.The risk score was correlated with clinical features.IFI16 was highly expressed in cancer tissues and associated with poor prognosis.Conclusion The prognostic model constructed in this study has good performance.Integrating clinical features, it can improve the accuracy of prognosis assessment of ccRCC.

ObjectiveTo observe the clinical efficacy and safety of lung-boosting moxibustion with different moxibustion doses in the treatment of stable chronic obstructive pulmonary disease （COPD）. MethodsA total of 240 stable COPD patients with lung qi deficiency， lung-spleen qi deficiency， or lung-kidney qi deficiency syndrome were enrolled， all receiving standardized western medical treatment and lung-boosting moxibustion. According to the thickness of the moxa wool， the thickness of the ginger paste， and the treatment duration， patients were randomly divided into eight groups， which were Group A （3.0 cm thickness moxa， 2.5 cm ginger mud， 60 min treatment time）， Group B （3.0 cm thickness moxa， 2.5 cm ginger mud， 90 min treatment time）， Group C （2.0 cm thickness moxa， 2.5 cm ginger mud， 60 min treatment time）， Group D （2.0 cm thickness moxa， 2.5 cm ginger mud， 90 min treatment time）， Group E （3.0 cm thickness moxa， 2.0 cm ginger mud， 60 min treatment time）， Group F （3.0 cm thickness moxa， 2.0 cm ginger mud， 90 min treatment time）， Group G （2.0 cm thickness moxa， 2.0 cm ginger mud， 60 min treatment time）， Group H （2.0cm thickness moxa， 2.0cm ginger mud， 90 min treatment time）. Treatment was performed once every 15 days for 3 months， with a total of six sessions. The primary outcomes were the scores of the clinical symptoms and signs questionnaire for stable COPD before and after treatment， comprising seven symptom domains （cough， sputum production， wheezing， chest tightness， shortness of breath， fatigue， and cyanosis） and the total score. The secondary outcomes included the scores of the modified COPD patient-reported outcome questionnaire （mCOPD-PRO） involving the physical， psychological， and environmental domains and the total score， the scores of the modified Effectiveness Satisfaction Questionnaire for COPD （mESQ-COPD） involving clinical symptoms， work and daily living capacity， environmental adaptability， treatment efficacy， and the total score before and after treatment， the moxibustion sensation scores （heat sensation score and special sensation intensity score） during treatment， and cutaneous temperature （the maximum temperature achieved， the duration of temperature maintained at ≥43 ℃， and the time required to reach 43 ℃）. The safety evaluation was carried out after treatment. ResultsDuring the study， 18 patients withdrew， and 222 patients were included in the per-protocol set for analysis， including 27 in Group A， 28 in Group B， 28 in Group C， 29 in Group D， 27 in Group E， 28 in Group F， 27 in Group G， and 28 in Group H. After treatment， the total score on the clinical symptoms and signs questionnaire， as well as the cough and chest tightness symptom scores in Group F were significantly lower than those in other groups （P＜0.05）. Group F demonstrated statistically lower total mCOPD-PRO score and physical domain score （P＜0.05）， lower total mESQ-COPD score and the scores for clinical symptoms and treatment efficacy （P < 0.05）， as well as higher heat sensation score （P＜0.05） than other groups. The maximum temperature was significantly higher in Group E and Group F than in the other groups， while the duration of temperature maintained at ≥43 ℃ was significantly longer in Group F than other groups， and the time required to reach 43 ℃ was significantly shorter in Group E and Group F than in Groups C， D， G， and H （P＜0.05）. No significant abnormalities were observed in routine blood tests， liver function， renal function， urinalysis， or electrocardiograms before and after treatment in any group. No serious adverse events occurred during the treatment period. ConclusionThere are differences in the efficacy of different moxibustion doses of lung-boosting moxibustion in the treatment of stable COPD. Moxibustion with 3.0 cm thickness moxa， 2.0 cm ginger mud， and 90 min treatment time can obtain relatively superior efficacy while demonstrating a favorable safety profile.

[Objective] To investigate the protective effect of Salvia miltiorrhiza injection (SMI) on the kidneys of HBOC-CHP01 resuscitated haemorrhagic shock rats. [Methods] A 50% haemorrhagic shock rat model was established, with 12 rats divided into two groups: SMI + HBOC-CHP01 group and HBOC-CHP01 group, with 6 rats in each group. The rats in the SMI+ HBOC-CHP01 group were given an equal volume of HBOC-CHP01 for resuscitation after haemorrhagic shock, and an 8 mL/kg dose of SMI. Rats in the HBOC-CHP01 group were resuscitated by administering an equilibrium blood loss volume of HBOC-CHP01 and given an 8 mL/kg dose of 0.9% NaCl solution. Blood was taken from rats at five points: before bloodletting (baseline), during haemorrhagic shock (HS), immediately after resuscitation (RS0h), 1 h after resuscitation (RS1h), and 24 h after resuscitation (RS24h). A blood gas analyser was used to detect the lactate level (Lac), glucose content (Glu), residual base (BEecf), pH, bicarbonate (HCO3-), high iron haemoglobin (MetHb). White blood cells (WBC), platelets (PLT), haemoglobin content (Hb), carboxyhaemoglobin (COHb) were detected using a quintuple classification. Blood creatinine (SCr), uric acid (UA), kidney-related indexes were detected using biochemistry instrument. Kidney tissues of the rats were taken after 24 h of resuscitation and after execution, and the inflammation of kidneys of the rats of the two groups was analyzed using HE staining. Fluorescence staining was used to detect the level of ROS in the kidneys of rats in both groups. [Results] At RS 0h, the Beecf, Glu and Lac levels of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group, and the pH level of rats in the SMI+HBOC-CHP01 group was significantly higher than that of rats in the HBOC-CHP01 group, and the Glu levels of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group at RS 1h. At RS 0h, the WBC, PLT and COHb contents of rats in the SMI+HBOC-CHP01 group were all significantly higher than those of rats in the HBOC-CHP01 group, and at RS 1h, the WBC content of rats in the SMI+HBOC-CHP01 group was significantly higher than that of rats in the HBOC-CHP01 group; at RS 1h, the UA content of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC-CHP01 group; at RS 24h, the SCr content of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC-CHP01 group; at RS 24h, the inflammation level of kidney tissues of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC -CHP01 group rats, and the ROS and MPO levels in the kidney tissues of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group. [Conclusion] The combination of Salvia miltiorrhiza injection during the resuscitation of rats with severe haemorrhagic shock by HBOC-CHP01 can alleviate renal injury by reducing inflammatory response and oxidative stress.

[Objective] To extract hemoglobin (Hb) from red blood cells using osmotic pressure swelling method, expected to achieve a hemoglobin dissolution rate of ≥80% and a cell membrane integrity rate of ≥70%. [Methods] Human umbilical cord blood red blood cells were used as raw materials and phosphate buffer solution was used as the swelling solution for red blood cells. A three factor three-level orthogonal experiment (n=3) was conducted to determine the optimal matching conditions for selecting the osmolality molar concentration of phosphate buffer solution, pH value of hypotonic phosphate buffer solution and volume ratio of hypotonic phosphate buffer solution to washed red blood cells. Red blood cell swelling solution samples (n=6) were prepared by the optimal matching conditions and the original process conditions. The hemoglobin dissolution rate and cell membrane integrity rate were checked. In the expanded comparative experiment, red blood cell swelling solution samples (n=6) were prepared by the optimal matching conditions and the original process conditions, which was filtered by ultrafiltration membranes. The filtration time and hemoglobin yield were checked. [Results] The optimal matching conditions for preparing red blood cell swelling solution were obtained through orthogonal experiment as follows: osmotic pressure molar concentration was 30 mOsmol/Kg, pH was 7.8, and phosphate buffer to red blood cell volume ratio was 6∶1. On the basis of the above conditions, the red blood cell swelling solution sample was compared with the original process sample: the hemoglobin dissolution rate was (82.4±1.8)% vs (78.6±3.0)% (P<0.05), and the cell membrane integrity rate was (65.8±4.0)% vs (28.7±2.3)% (P<0.05). In the expanded comparative experiment, the optimal matching conditions were compared with the original process conditions: filtration time(s) (327±9) vs (434±13) (P<0.05), and hemoglobin yield was (72.3±1.2)% vs (66.0±1.4)% (P<0.05). [Conclusion] Compared with the original preparation process, the hemoglobin extraction process which optimized through orthogonal experiments greatly reduces the cell membrane fragmentation rate and minimizes the entry of cell membrane matrix into the target solution, ensuring a slightly higher hemoglobin dissolution rate, and reducing the preparation difficulty for the subsequent cell membrane separation and further purification.

Head and neck squamous cell carcinoma(HNSCC)is the sixth most common malignant tumor in the world.Due to its high incidence,poor prognosis,relapse-prone characteristics,and the existence of treatment resist-ance,HNSCC has a serious impact on patients'daily lives.As a bridge between innate immunity and adaptive immu-nity,dendritic cell(DC)plays a crucial role in tumor progression.In recent years,numerous studies have found that DC has complex and diverse functions in the tumor microenvironment(TME),and has a significant impact on the oc-currence and development of HNSCC.In this article,we provide a review of the mechanism of DC in HNSCC TME.

Objective:To present a set of clinically representative quality assurance (QA) test cases for stereotactic radiosurgery (SRT) plans of multiple intracranial metastases, in order to assess the plan quality and machine execution capabilities.Methods:Based on the clinical characteristics of multiple brain metastases, four groups of test cases with three target volumes (TVs), six TVs, nine TVs, and TVs near organs at risk (OARs) were designed. For these cases, SRT plans were developed, and plan quality was assessed using metrics including the Radiation Therapy Oncology Group conformality index (RTOG CI), gradient index (GI), homogeneity index (HI), and the volume of normal brain tissue receiving a dose of 24 Gy ( V24 Gy), which was defined as the volume enclosed by the 24 Gy isodose line around the Brain-PTV ( V24 Gy of Brain-PTV). Verification plans were generated for each test case, including the verification of point doses, planar doses (PD), and SRS MapCHECK (SMC) semiconductor matrix planar doses. Compared with the calculated result of the treatment planning system (TPS), the criteria for the γ analysis of planar doses were set at 1 mm/2% and 2 mm/2%. Results:For the four groups of test cases, the mean CI, GI, HI, and V24 Gy of Brain-PTV were 1.04±0.03, 3.79±0.40, 0.73±0.01 and (7.46±3.80) cm 3, respectively. The mean deviations of the point doses were 0.88%±0.98%, 1.47%±0.79%, 1.52%± 0.76%, and 1.17% ± 0.38%, respectively. The mean γ passing rates of the single fields for PDs were greater than 98% at 2 mm/2% and exceeding 96% at 1 mm/2%, and the mean γ pass rates of the SMC semiconductor matrix for PDs were 97.75% ± 2.31% and 99.33% ± 0.62%, at 1 mm/2% and 2 mm/2% respectively. Conclusions:The proposed QA test cases for SRT of multiple intracranial metastases allow for the effective assessments of the plan quality and machine execution capabilities and, thus, can assist various centers in clinical applications.

Objective:To establish a fluorescent quantitative RT-PCR assay based on N_sgRNA of SARS-CoV-2 and preliminarily apply it on real samples.Methods:Recombinant plasmid, specific primers and probes of N_sgRNA were designed and synthesized based on Wuhan-Hu-1/2019_MN908947 and synthesis mechanism of subgenomic RNA (sgRNA). Using recombinant plasmid as amplification templates, the optimal reaction conditions and reaction system were screened according to the Ct value, fluorescence intensity, and shape of amplification curve and was evaluated for sensitivity, reproducibility, and specificity. Meanwhile, the possibility of practical application of the method was explored by testing 172 clinical samples and 256 municipal wastewater samples. Results:A qRT-PCR assay for N_sgRNA in SARS-CoV-2 was initially established. The detection limit of the assay was 20 copies/mL, and the variation coefficients of in-batch (0.002%~0.767%) and batch to batch repetition (0.016%~0.752%) were less than 1%. Only N_sgRNA recombinant plasmid was detected in the specificity assay. So the method is more highly sensitive, specific and reproducible. The RatiosgRNA/ gRNA of clinical samples HK.3, EG.5.1, JN.1 and their sub-lineages and their corresponding urban sewage samples in epidemic period were significantly different ( P<0.05). There is a strong correlation between the median of RatiosgRNA/ gRNA in clinical samples and sewage samples in the same period (correlation coefficient r=1.000, P=0.010). Conclusions:In this study, a qRT-PCR method for detecting N_sgRNA of SARS-CoV-2 was established and the method has the characteristics of higher sensitivity, stronger specificity and better repeatability, and it can be used to detect SARS-CoV-2 infectivity.