Objective: To develop a RHCE genotyping assay based on kompetitive allele-specific PCR (KASP) and assess its clinical accuracy for RhCE blood group determination. Methods: KASP primers were designed to interrogate three RHCE loci: the 109 bp insertion/deletion in intron 2, c. 307T>C, and c. 676C>G. A total of 1 194 RhD-positive inpatients from Chengdu were typed by both KASP genotyping and manual tube serology. Discordant samples (n=10) were retested by both methods and further resolved by Sanger sequencing. An additional 377 cases were tested for the c. 48C>G locus to evaluate the predictive accuracy of individual loci and combined locus testing for RhC antigen. Results: Genotyping concordance with serology was 100.0% for both the c. 676C>G locus (RhE/Rhe) and the c. 307T>C locus (Rhc). For RhC prediction using the 109 bp insertion, overall accuracy was 99.7% (1 191/1 194); the 3 discordant cases were confirmed by Sanger sequencing to be false negatives attributable to 109 bp deletion in intron 2. Testing the c. 48C>G allele for RhC prediction yielded 7 false positives, with an accuracy of 98.1% (370/377). RhC antigen status was determined by combining the 109 bp insertion and the c. 48C allele. After excluding 10 samples with inconsistent results between the two loci, the accuracy reached 100% in the remaining 367 samples. When both loci were applied in combination, accuracy reached 100% in the 367 cases with concordant results. Among the 1 194 patients, CCee (45.8%) and CcEe (31.7%) were the most common RhCE phenotypes. The e antigen had the highest positivity rate (92.2%), and the Ce haplotype was the most frequent (66.9%). Conclusion: The KASP-based RHCE genotyping method achieves high accuracy for clinical RhCE typing. Combining the 109 bp insertion/deletion with the c. 48C allele significantly improves RhC antigen prediction compared with either locus alone. This method was applied to RhCE genotyping of 1 194 RhD-positive inpatients in Chengdu, providing local RhCE phenotype and haplotype distribution data to support RhCE-matched transfusion practice.

[Purpose]To analyze the incidence and mortality of lung cancer in Zhejiang cancer regi-stration areas in 2021 and trends from 2000 to 2021.[Methods]Using data from Zhejiang cancer registration areas from 2000 to 2021,the crude rate,age-standardized rate by Chinese standard population(ASRC)and the world standard population(ASRW),and composition ratio were calcu-lated to describe the incidence and mortality in 2021.Joinpoint regression was employed to ana-lyze temporal trends over the 22 years,calculating the annual percentage change(APC),average annual percentage change(AAPC),and their 95％confidence intervals(CI).[Results]The crude incidence rate of lung cancer in 2021 was 128.58/105,with an ASRC of 68.74/105,ac-counting for 24.22％of all cancers;the crude mortality rate was 51.55/105,with an ASRC of 21.58/105,constituting 27.98％of all cancer deaths.Both crude incidence rate(AAPC=5.73％,P＜0.001)and ASIRC(AAPC=4.14％,P＜0.001)of lung cancer showed significant increasing trends from 2000 to 2021.The crude mortality rate increased slowly(AAPC=1.76％,P＜0.001),while the ASMRC(AAPC=-0.40％,P=0.035)exhibited a modest decline.The increase in incidence was more pronounced in females than males,though no statistically significant difference was observed in mortality trends.The incidence rates in urban and rural areas showed similar growth trends;the crude mortality rate in rural areas was growing significantly faster than that in urban areas,and the decline in the ASRC was only statistically significant in urban areas.The incidence rate was in-creasing in all age groups.The mortality rate was declining among younger people(＜65 years old).The age group of 15～44 years old demonstrated a greater increase in incidence and a sharper decline in mortality than those aged 45 years old and above.[Conclusion]From 2000 to 2021,Zhejiang Province experienced a marked rise in lung cancer incidence with escalating disease burden.Despite severe population aging,ASRC of mortality declined steadily,reflecting effective prevention and control measures.

[Purpose]To analyze the incidence and mortality of lung cancer in Zhejiang cancer regi-stration areas in 2021 and trends from 2000 to 2021.[Methods]Using data from Zhejiang cancer registration areas from 2000 to 2021,the crude rate,age-standardized rate by Chinese standard population(ASRC)and the world standard population(ASRW),and composition ratio were calcu-lated to describe the incidence and mortality in 2021.Joinpoint regression was employed to ana-lyze temporal trends over the 22 years,calculating the annual percentage change(APC),average annual percentage change(AAPC),and their 95％confidence intervals(CI).[Results]The crude incidence rate of lung cancer in 2021 was 128.58/105,with an ASRC of 68.74/105,ac-counting for 24.22％of all cancers;the crude mortality rate was 51.55/105,with an ASRC of 21.58/105,constituting 27.98％of all cancer deaths.Both crude incidence rate(AAPC=5.73％,P＜0.001)and ASIRC(AAPC=4.14％,P＜0.001)of lung cancer showed significant increasing trends from 2000 to 2021.The crude mortality rate increased slowly(AAPC=1.76％,P＜0.001),while the ASMRC(AAPC=-0.40％,P=0.035)exhibited a modest decline.The increase in incidence was more pronounced in females than males,though no statistically significant difference was observed in mortality trends.The incidence rates in urban and rural areas showed similar growth trends;the crude mortality rate in rural areas was growing significantly faster than that in urban areas,and the decline in the ASRC was only statistically significant in urban areas.The incidence rate was in-creasing in all age groups.The mortality rate was declining among younger people(＜65 years old).The age group of 15～44 years old demonstrated a greater increase in incidence and a sharper decline in mortality than those aged 45 years old and above.[Conclusion]From 2000 to 2021,Zhejiang Province experienced a marked rise in lung cancer incidence with escalating disease burden.Despite severe population aging,ASRC of mortality declined steadily,reflecting effective prevention and control measures.

Objective To investigate the effect and mechanism of cinobufotalin in inhibiting hepatocellular carcinoma(HCC)metastasis by regulating epithelial-mesenchymal transition(EMT).Methods A total of 36 male BALB/c nude mice,aged 6 weeks,were given injection of MHCC97H cells via the caudal vein to establish a model of HCC lung metastasis,and then the mice were randomly divided into high-and low-dose cinobufotalin groups and control group.Since the day of modeling,the mice in the high-and low-dose cinobufotalin groups were given intraperitoneal injection of cinobufotalin at a dose of 120 μL/kg and 60 μL/kg,respectively,and those in the control group were given intraperitoneal injection of normal saline,twice a week.After 8 weeks,HE staining was performed for lung tissue to measure the lung metastasis rate of HCC.MHCC97H cells were treated with high-dose(2.5 μL/mL)or low-dose(5 μL/mL)cinobufotalin for 24 hours,and wound healing assay,RT-PCR,and Western blot were used to measure cell migration ability and the expression of EMT-related molecules.MHCC97H cells were induced in a simulated hypoxic environment with CoCl2 incubation,with high-and low-dose cinobufotalin added for intervention,and wound healing assay and Western blot were used to investigate the effect of cinobufotalin on cell migration ability and EMT induced by hypoxia.Transcriptome analysis was used to investigate the effect mechanism of cinobufotalin on MHCC97H cells,and Western blot was used to observe the effect of cinobufotalin on the expression levels of protein kinase B(AKT)and phosphorylated AKT(P-AKT)in MHCC97H cells.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups;the independent-samples t test was used for comparison of categorical data between two groups.Results Compared with the control group,the cinobufotalin group had a significant reduction in the lung metastasis rate of HCC.Compared with the control group,cinobufotalin intervention reduced the wound healing rate of MHCC97H cells,upregulated the expression of epithelial-type molecules(t=2.860,P<0.05),and downregulated the expression of EMT transcription factors(EMT-TFs)and mesenchymal molecules(t=3.545,2.022,2.852,and 2.341,all P<0.05).Hypoxia induction upregulated the wound healing rate of MHCC97H cells and the expression levels of mesenchymal molecules and EMT-TFs(P<0.05),and cinobufotalin intervention reversed EMT change and inhibited wound healing(P<0.05).The transcriptome analysis of MHCC97H cells showed significant gene differences between the cinobufotalin group and the control group,and cinobufotalin mainly affected the expression of genes associated with tumor,metabolism,immunity,and signal transduction,with the largest number of differentially expressed genes in the AKT signal transduction pathway.Further measurement showed that cinobufotalin intervention downregulated the expression levels of AKT,P-AKT,and P-AKT/AKT in MHCC97H cells(t=2.434,3.401,and 2.258,all P<0.05).Conclusion Cinobufotalin can inhibit the metastasis of HCC,especially hypoxia-induced HCC metastasis,and regulation of EMT mediated by the AKT signal transduction pathway in HCC cells might be one of its mechanisms of action.

Objective:To investigate the clinical efficacy of non-drug interventions in hospice and palliative care(HPC)for improving cancer-related fatigue(CRF)in elderly individuals and its impact on quality of life.Methods:This study presents findings from a single-center randomized controlled trial conducted at Zhejiang Hospital, focusing on 40 elderly patients experiencing cancer-related fatigue(CRF)between February 2022 and February 2023.The participants were randomly assigned into a control group and an intervention group, each consisting of 20 individuals, using the random number table method.Both groups received routine comprehensive treatment, with the intervention group additionally receiving hospice and palliative care(HPC)non-drug intervention.Following 6 weeks of continuous treatment, the study compared the clinical efficacy, changes in CRF, and quality of life before and after treatment between the two groups.Results:Comparing the baseline data of the two groups of patients, the difference was not statistically significant(all P>0.05).After 6 weeks of treatment, patients in the intervention group reported lower levels of current fatigue, general fatigue, worst fatigue in the past 24 hours, and impact of fatigue on various aspects of their lives compared to the control group(all P<0.01).The clinical remission rate of cancer-related fatigue(CRF)in the intervention group was 60%, significantly higher than the 5% in the control group( P<0.01).Additionally, the intervention group showed improvement in overall quality of life and emotional function with decreased symptom areas scores(fatigue, nausea and vomiting, shortness of breath, sleep disorders, and loss of appetite)( P<0.01 for quality of life and emotional function, P<0.05 for symptom areas). Conclusions:Non-pharmacological interventions within the context of hospice and palliative care have been shown to alleviate cancer-related fatigue in elderly cancer patients, ultimately improving their quality of life.These interventions have demonstrated positive effects on various aspects such as overall quality of life, functional status, fatigue, nausea and vomiting, shortness of breath, sleep disorders, and decreased appetite.Furthermore, these interventions are considered safe and effective in the treatment of elderly cancer patients experiencing fatigue.

Objective:To observe the clinical efficacy of body acupuncture combined with FANG's scalp acupuncture in treating limb dysfunction in the remission stage of stroke and explore the mechanism. Methods:Sixty patients in the remission stage of stroke with limb dysfunction were divided into a control group and a treatment group using the random number table method,with 30 cases in each group.The control group was treated with ordinary acupuncture based on the conventional rehabilitation treatment,and the treatment group was additionally offered FANG's scalp acupuncture.The interventions were conducted once daily with 10 sessions as 1 course.The Fugl-Meyer assessment(FMA)and Barthel index(BI)scores and serum malondialdehyde(MDA)and superoxide dismutase(SOD)levels were compared after 2 treatment courses,and the clinical efficacy was evaluated. Results:After treatment,the FMA and BI scores increased in both groups(P<0.05)and were higher in the treatment group than in the control group(P<0.05).Both groups showed a decreased MDA level and an increased SOD level after the intervention,all showing statistical significance(P<0.05);there were significant differences between the two groups(P<0.05). Conclusion:Based on the conventional rehabilitation treatment,ordinary acupuncture used alone or in combination with FANG's scalp acupuncture can lower the oxidative stress level and improve limb function in treating limb dysfunction in the remission stage of stroke;body acupuncture plus FANG's scalp acupuncture can produce better results.

Objective:To investigate the relationship between vaccination doses and the duration of positive nucleic acid or antigen tests in Corona Virus Disease 2019 (COVID-19) patients during omicron epidemic.Methods:A cross-sectional study was conducted from December 26 to December 31, 2022 among COVID-19 patients from all community health service centers in 16 districts of Beijing municipality selected by multi-stage stratified cluster quota sampling method. A questionnaire was used to collect the basic information, vaccination doses, results of nucleic acid or antigen tests of all subjects. The subjects were divided into 5 groups according to the doses of COVID-19 vaccination: the non-vaccination group, the 1, 2, 3 and 4 doses vaccination group. The relationship between vaccination doses and the duration of positive nucleic acid or antigen tests was analyzed by univariate and multivariate logistic regression models.Results:A total of 10 506 COVID-19 cases were included in the study with the age of (43.3±13.7) years. The duration of positive nucleic acid or antigen tests was longer than 7 days in 59.4%(276/465), 51.5%(67/130), 50.6%(355/701), 46.1% (3 464/7 520) and 39.2%(662/1 690) of non-vaccination, and 1, 2, 3, 4 dose vaccination groups, respectively ( P<0.001). Multivariate logistic regression analysis showed that compared with non-vaccination patients, the vaccination dose was an independent protective factor for duration of positive nucleic acid or antigen tests>7 days, and the OR values were 0.7 (95% CI 0.5-0.9, P=0.015), 0.6 (95% CI 0.5-0.8, P<0.001) and 0.5 (95% CI 0.4-0.6, P<0.001) for 2, 3 and 4 doses of vaccination, respectively. Conclusion:The vaccination doses are independently related to the duration of positive nucleic acid or antigen tests in COVID-19 patients and the risk is gradually decreases with the increasing vaccination doses.

Objective:To seek specific response points on the body surface of patients with primary dysmenorrhea(PD)by observing blood perfusion unit(PU)at different points of the three Yin meridians of foot using laser speckle contrast imaging(LSCI). Methods:Eighty PD patients were recruited as a PD group,and 80 healthy female undergraduates were taken as a normal group.During one menstrual cycle(before menstruation,during menstruation,and 3 d after menstruation),each participant was examined using the LSCI system to determine PU at bilateral Taixi(KI3),Taibai(SP3),Taichong(LR3),Shuiquan(KI5),Diji(SP8),Zhongdu(LR6),Sanyinjiao(SP6),and Xuehai(SP10)and non-acupuncture points.The researchers in charge of point location,operation,and statistical analysis were not aware of grouping.PU at the detection spots was taken as the outcome measure. Results:Compared with the normal group,the PD group showed increases in PU at right Taixi(KI3)before menstruation(P<0.05)and at bilateral Zhongdu(LR6)and right Diji(SP8)during menstruation(P<0.05).At the other time points,significance was not found between the two groups in comparing PU at the detected spots. Conclusion:Compared with healthy participants,PD patients present specific changes in PU at Taixi(KI3),Diji(SP8),and Zhongdu(LR6)at specific time points during the menstrual cycle,which provides a reference for acupuncture-moxibustion treatment of PD in clinical settings.

Objective:To understand the epidemic situation of HIV/AIDS and its change trend in the Belt and Road countries and provide evidence for the improvement of prevention and control of the cross-border transmission of HIV/AIDS.Methods:The 145 countries that have signed the cooperation document of the Belt and Road initiative with China were selected in our study. Age-standardized incidence rate and prevalence rate of HIV/AIDS in the 145 countries from 2013 to 2019 were downloaded from the Global Burden of Disease Study 2019. Age-standardized incidence rate and prevalence rate of HIV/AIDS in 2019 were used to describe the HIV/AIDS epidemics in 145 countries, and the estimated annual percentage change (EAPC) of incidence was calculated to analyze the trend of HIV/AIDS incidence from 2013 to 2019.Results:In 2019, Africa had the highest proportion of countries with HIV/AIDS incidence exceeding 40.00 per 100 000 (56.86%, 29/51), and Asia had the lowest proportion (5.41%, 2/37). The countries with the prevalence rate of HIV/AIDS exceeding 100.00 per 10 000 were almost distributed in Africa, accounting for 20.69% (30/145). From 2013 to 2019, the incidence rate of HIV/AIDS increased in 50 countries, accounting for 34.48% (50/145). The incidence rate of HIV/AIDS showed downward trends in 69 countries (47.59%, 69/145), and showed no significant change in 26 countries (17.93%, 26/145). The most obvious increase of incidence rate of HIV/AIDS was observed in Comoros (EAPC=15.60, 95% CI: 5.84-26.26) and the most obvious decrease was observed in Burundi (EAPC=-14.27, 95% CI: -15.21 to -13.31). Conclusions:In the Belt and Road countries, the most severe disease burden of HIV/AIDS was observed in countries in Africa, and the incidences of HIV/AIDS increased rapidly in some European countries, which means the risk of cross-border transmission still exists. Hence, the prevention and control of HIV/AIDS in China should be further strengthened in the future.