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: Dual anti–human epidermal growth factor receptor 2 (HER2) drugs have become the standard regimen for neoadjuvant systemic treatment (NST) to HER2-positive breast cancer patients. However, the efficacy varies greatly among patients with different HER2 protein expression levels. Materials and Methods: A total of 575 HER2-positive breast cancer patients from multiple centers throughout China from 2013 to 2022 were retrospectively analyzed. We compared clinicopathological features in different HER2 immunohistochemistry classes (HER2 2+/in situ hybridization [ISH] + or HER2 3+), and their difference in response to NST and survival with single or dual anti-HER2 drugs. Drug sensitivity assays were used to evaluate different efficacy of anti-HER2 drugs in vitro. Results: Compared to HER2 3+ subgroup, the HER2 2+/ISH+ group had a higher proportion of hormone receptor–positive status (48.7% vs. 76.1%, p < 0.001), more HER2 protein loss after NST, lower pathological complete response (pCR) rate (46.07% vs. 16.24%, p < 0.001), and tended to have worse disease-free survival (DFS). In HER2 2+/ISH+ patients, treated with pertuzumab and trastuzumab in combination had no significant improvement in pCR (19.12% vs. 12.24%, p=0.287) and DFS (p=0.908) than using alone. Drug sensitivity assay showed poor efficacy with dual anti-HER2 drugs in HER2 2+/ISH+ cell lines; however, fam-trastuzumab deruxtecan drugs had a satisfactory effect. Conclusion Owing to the differences in clinicopathological features and treatment efficacy, we considered the HER2 2+/ISH+ group to be a distinct subtype and defined it as the HER2-moderate–positive subgroup. In this subgroup, dual anti-HER2 drugs did not exert significant improvement in pCR and DFS. Therefore, treatment optimization is warranted, with antibody-drug conjugate drugs as potential options.

[Objective]To summarize Professor ZHANG Yonghua's clinical experience in treating emotional-type insomnia based on the"wind movement smoothening the Xuanfu(pores)"theory for clinical reference.[Methods]Through learning,case collection,literature review and combining modern medical mechanisms,the article explains Professor ZHANG's understanding of the pathogenesis of emotional-type insomnia,explores the syndrome differentiation methods and treatment approaches based on the"wind movement smoothening the Xuanfu"theory,and provides a clinical case for verification.[Results]The core pathogenesis of insomnia caused by emotional disorders is the blockage of the"Xuanfu".Professor ZHANG points out that patients with emotional disorders suffer from the blockage of the"Xuanfu"due to the disharmony of Qi movement,internal disturbance of depressed fire,deficiency of Qi and blood,and accumulation of phlegm and blood stasis,which lead to prolonged and difficult-to-treat insomnia.His treatment approach often starts from"unblocking the Xuanfu",advocating the basic therapeutic method of"wind movement smoothening Xuanfu",using wind herbs to relieve depression and regulate Qi,disperse fire and clear heat,tonify deficiency and nourish the spirit,remove phlegm and resolve blood stasis to unblock the Xuanfu,and harmonize Yin and Yang to restore balance.The attached medical case was a patient of insomnia due to dissociative conversion disorder,whose main pathogenesis was the blockage of the Shaoyang pivot and the internal hyperactivity of the heart fire of the Shaoyin,presenting the syndrome of depressed gallbladder and phlegm-heat.The treatment approach was to resolve phlegm and clear heat,subdue the floating Yang,further regulate the exterior and interior,harmonize Yin and Yang,and unblock the Xuanfu to relieve depression,so as to enable the patient to sleep peacefully.[Conclusion]Professor ZHANG's treatment of emotional-type insomnia based on the"wind movement smoothening the Xuanfu"theory is highly effective and offers a new approach to treating insomnia with traditional Chinese medicine,which is worth promoting and applying.

[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 discuss the efficacy of treating allergic rhinoconjunctivitis(ARC)with intranasal acupuncture at Neiyingxiang(EX-HN9)and Biqiu(Extra)points and the effects on inflammatory cells.Methods:A total of 207 ARC patients were recruited and divided into an observation group(104 cases)and a control group(103 cases)using the random number table method.The control group was prescribed desloratadine dispersible tablets and azelastine hydrochloride eye drops for 1 week,and the observation group was given additional acupuncture at bilateral Neiyingxiang(EX-HN9)and Biqiu(Extra)points,once daily consecutively over 1 week.The clinical efficacy was compared between the two groups when the treatment finished.Before and after treatment,both groups were scored using the rhinoconjunctivitis quality of life questionnaire(RQLQ),and the levels of blood eosinophils(EOS)and interleukin(IL)-17 were detected.The cured cases were observed for recurrence rate at the 6-month follow-up.Results:The observation group had more significant clinical efficacy than the control group(P<0.05).After treatment,The RQLQ score declined in both groups(P<0.05)and was lower in the observation group than in the control group(P<0.05).After the intervention,the levels of blood EOS and IL-17 dropped in the two groups and were lower in the observation group than in the control group(P<0.05).At the 6-month follow-up among the cured cases,a lower recurrence rate was found in the observation group compared to the control group(P<0.05).Conclusion:Based on conventional medications,acupuncture at Neiyingxiang(EX-HN9)and Biqiu(Extra)points can notably enhance the clinical efficacy in treating ARC patients,reducing the inflammatory cells and lowering the recurrence rate.

[Objective]To analyze the therapeutic characteristics of Zhuyu pill in the Handbook of Prescriptions for Emergency.[Methods]This article used the methods of in-depth investigation for the origin of diseases,through discussion of prescriptions,analysis of their compositions and clinical practice.It analyzed five aspects of the Zhuyu pill in Handbook of Prescriptions for Emergency including formulation identification,compilation and supplementation of texts,pathogenesis of diseases and syndromes,explanation of ingredients,and the correlation between syndrome identification and treatment strategies,with case studies provided as example.[Results]The Zhuyu pill in the Handbook of Prescriptions for Emergency possesses the functions of warming the kidneys and invigorating the spleen,dissipating cold and relieving pain,descending counter-flow of Qi and resolving phlegm,nourishing blood and promoting digestion.This formula is effective in treating diseases such as chest impediment,heart pain,as well as Taiyin syndrome,and others.Clinically,it is primarily indicated for conditions characterized by symptoms like heart pain,sour taste in mouth,indigestion,yellowish complexion,abdominal distension,and a slow or feeble pulse.The case reported involved diagnoses of chest impediment,heart pain and phlegm-fluid retention,all showing remarkable efficacy with Zhuyu pill and its modifications.[Conclusion]The Zhuyu pill in the Handbook of Prescriptions for Emergency can effect in treating chest impediment,heart pain,Taiyin syndrome,as well as internal and external phlegm retention and obstruction,thus restoring the normal circulation of Qi,blood and fluids.

[Objective]To summarize Professor CHEN Jian's clinical experience in treating pediatric upper airway cough syndrome,which is based on the"Xuanfu"theory,and to learn how to use wind-dispelling herbs.[Methods]Through clinical observation,medical case organization,literature search and integration of clinical practice experience,this paper elaborates Professor CHEN's understanding of the etiology and pathogenesis of upper airway cough syndrome and summarizes the clinical experience of treating this disease from the perspective of"opening Xuanfu with wind-dispelling herbs",in addition with relevant cases for verification.[Results]Professor CHEN considers the lurking wind in the"Xuanfu"and the upward reversal of lung Qi as the basic pathogenesis when diagnosing and treating pediatric upper airway cough syndrome.The deficiency of Zhengqi and the lack of nourishment in the"Xuanfu"are seen as the root causes of the disease.The obstruction of"Xuanfu"with phlegm,dampness and blood stasis are the pathogenic factors.The prescription is based on"Jiuke"Recipe,with wind-dispelling herbs being a continuous thread throughout.The fundamental treatment approach is using wind-dispelling herbs to treat wind disease,and to open and consolidate the"Xuanfu".The application of wind-dispelling herbs aims to expel wind,open"Xuanfu",overcome dampness and facilitate the flow of"Xuanfu",as well as support and nourish"Xuanfu".The treatment makes differentiation between the nasal and pharyngeal areas,with simultaneous treatment of the lung orifices,ensuring the smooth flow of"Xuanfu"and harmony of the viscera.A case was presented at the end of the paper to illustrate the treatment principle of dispelling wind and opening"Xuanfu",combining both attack and supplementation,leading to the unblocking of"Xuanfu",the expulsion of lurking wind,the transformation of phlegm and dampness and the self-elimination of cough,with a definite therapeutic effect.[Conclusion]Professor CHEN's approach to treating pediatric upper airway cough syndrome with"wind-dispelling herbs to open the Xuanfu"is effective,which provides insights and experience for clinical diagnosis and treatment of the disease,which has significant enlightening significance.

Objective:To observe the clinical efficacy of acupuncture combined with cognitive training in treating mild-to-moderate poststroke cognitive impairment and the impact on cognitive function and cerebral blood flow.Methods:Eighty-eight participants with mild-to-moderate cognitive impairment after a stroke were allocated to an observation group and a control group using the random number table method,each consisting of 44 cases.The two groups received the same conventional symptomatic treatment.In addition,the control group was intervened by cognitive training,and the observation group by cognitive training and Tong Du Xing Shen(unblocking the Governor Vessel and awakening the mind)acupuncture.The intervention lasted 4 weeks in both groups.By the end of the intervention,the two groups were observed for clinical efficacy and the improvements in the symptom score of traditional Chinese medicine(TCM),cognitive function[mini-mental state examination(MMSE)and Montreal cognitive assessment(MoCA)],cerebral blood flow velocities[cerebral vascular resistance index(RI),arterial blood flow mean velocity(Vm),and arterial systolic velocity(Vs)],and serum biomarkers of neurological deficit[neuron-specific enolase(NSE)and S100 β protein(S100β)].Results:During the observation,each group had 2 eliminated cases.After the intervention,the observation group achieved a total effective rate of 90.5%,higher than that in the control group,which was 73.8%(P<0.05);increases were found in the MMSE and MoCA scores in both groups(P<0.05),and the observation group was higher than the control group(P<0.05);in contrast,decreases were found in the levels of RI and serum NSE and S100β(P<0.05),and the observation group was lower than the control group(P<0.05);the Vs and Vm levels increased in both groups(P<0.05)and were higher in the observation group than in the control group(P<0.05).Conclusion:Tong Du Xing Shen acupuncture plus cognitive training is effective in treating mild-to-moderate poststroke cognitive impairment and can enhance the patient's cognitive function;improving cerebral blood circulation and reducing cerebral nerve damage may be involved in the mechanism.

Objective:To observe the clinical efficacy of DING's Tuina(Chinese therapeutic massage)plus Western medication in treating rotator cuff injury(RCI)and its impact on pain,shoulder function,and shoulder muscle strength in the patients.Methods:Sixty-two RCI patients were divided into an observation group and a control group using a simple randomization method,each consisting of 31 cases.Both groups of patients were prescribed oral celecoxib capsules.In addition,DING's Tuina treatment was offered to the observation group.After 6 weeks of treatment,the two groups were observed for changes in the Constant-Murley score(CMS)for shoulder joint function,the short-form McGill pain questionnaire(SF-MPQ)score,and peak torque(PT)and peak torque to body mass ratio(PT/BM)at angular velocities 120 °/s and 90 °/s as well as the peak torque ratio(PTR)of agonist muscles to antagonist muscles[including adductors to abductors(ADD/ABD)and flexors to extensors(F/E)].Results:After treatment,both groups presented decreased SF-MPQ scores and increased CMS,and the intra-group differences were statistically significant(P<0.05);the observation group had a lower SF-MPQ score and a higher CMS than the control group,showing statistical significance(P<0.05).After the intervention,the ADD/ABD decreased in the control group at the angular velocity 120 °/s in abduction,and the intra-group change was statistically significant(P<0.05);in the observation group,the PT and PT/BM increased compared to the baseline at angular velocities 120 °/s and 90 °/s in abduction,adduction,forward flexion,and backward extension,showing statistical significance(P<0.05).After treatment,the PT and PT/BM at angular velocities 120 °/s and 90 °/s and ADD/ABD at 120 °/s were higher in the observation group than in the control group,and the between-group differences were statistically significant(P<0.05).Conclusion:DING's Tuina combined with oral celecoxib capsules can reduce pain,improve the joint's motor function,and enhance the strength of shoulder muscle groups in RCI patients,and this treatment has a positive impact on the stability when performing shoulder joint abduction and adduction.