1.Effect of Liuwei Dihuangwan on EMT and Expression of CSC Properties in 4T1 Cells by Regulating Myeloid-derived Suppressor Cells
Lixiang ZHENG ; Ling HUANG ; Huiwen GUO ; Biyao GONG ; Xiaoying REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):1-10
ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum (LDP) on epithelial-mesenchymal transition (EMT) and the expression of cancer stem cell (CSC) properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells (MDSCs). MethodsSPF-grade female SD rats were randomly divided into three groups, which were given 0.39, 1.94, 3.89 g·kg-1·d-1 suspension of Liuwei Dihuangwan for 7 days, respectively, to prepare low-, medium-, and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 (CCK-8) assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry (FC). The migration, invasion, and proliferation of 4T1 cells were detected by cell scratch assay, Transwell invasion assay, and plate colony-forming assay, respectively. Western blot (WB) was used to detect the protein expression of transforming growth factor-β (TGF-β), nuclear factor-κB (NF-κB), C-X-C motif chemokine ligand 2 (CXCL2), E-cadherin, and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence (IF). ResultsCompared with the normal group, LDP showed no significant inhibitory effect on the cell viability of 4T1 cells, but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs, as well as the expression of TGF-β (P<0.05, P<0.01). The migration, invasion, and proliferation of 4T1 cells were increased after co-culture with MDSCs (P<0.05, P<0.01). The expressions of NF-κB, CXCL2, and N-cadherin and the proportion of CSC (CD44+CD24-) were elevated (P<0.05, P<0.01), while the expression of E-cadherin was decreased (P<0.05). After the intervention of MDSCs with LDP, followed by co-culture with 4T1 cells, the migration, invasion, and proliferation of 4T1 cells were obviously reduced (P<0.01). The expressions of NF-κB, CXCL2, and N-cadherin were decreased (P<0.05, P<0.01), and the expression of E-cadherin was increased (P<0.05, P<0.01). There was no statistical difference in the proportion of CSC (CD44+CD24-) in 4T1 cells. However, the proportion of CSC (CD44+CD24-) was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention (P<0.05, P<0.01). ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β, NF-κB, and CXCL2, reverse EMT, and reduce the characteristic expression of CSC to inhibit the migration, invasion, and proliferation of 4T1 cells.
2.Effect of Liuwei Dihuangwan on EMT and Expression of CSC Properties in 4T1 Cells by Regulating Myeloid-derived Suppressor Cells
Lixiang ZHENG ; Ling HUANG ; Huiwen GUO ; Biyao GONG ; Xiaoying REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):1-10
ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum (LDP) on epithelial-mesenchymal transition (EMT) and the expression of cancer stem cell (CSC) properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells (MDSCs). MethodsSPF-grade female SD rats were randomly divided into three groups, which were given 0.39, 1.94, 3.89 g·kg-1·d-1 suspension of Liuwei Dihuangwan for 7 days, respectively, to prepare low-, medium-, and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 (CCK-8) assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry (FC). The migration, invasion, and proliferation of 4T1 cells were detected by cell scratch assay, Transwell invasion assay, and plate colony-forming assay, respectively. Western blot (WB) was used to detect the protein expression of transforming growth factor-β (TGF-β), nuclear factor-κB (NF-κB), C-X-C motif chemokine ligand 2 (CXCL2), E-cadherin, and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence (IF). ResultsCompared with the normal group, LDP showed no significant inhibitory effect on the cell viability of 4T1 cells, but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs, as well as the expression of TGF-β (P<0.05, P<0.01). The migration, invasion, and proliferation of 4T1 cells were increased after co-culture with MDSCs (P<0.05, P<0.01). The expressions of NF-κB, CXCL2, and N-cadherin and the proportion of CSC (CD44+CD24-) were elevated (P<0.05, P<0.01), while the expression of E-cadherin was decreased (P<0.05). After the intervention of MDSCs with LDP, followed by co-culture with 4T1 cells, the migration, invasion, and proliferation of 4T1 cells were obviously reduced (P<0.01). The expressions of NF-κB, CXCL2, and N-cadherin were decreased (P<0.05, P<0.01), and the expression of E-cadherin was increased (P<0.05, P<0.01). There was no statistical difference in the proportion of CSC (CD44+CD24-) in 4T1 cells. However, the proportion of CSC (CD44+CD24-) was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention (P<0.05, P<0.01). ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β, NF-κB, and CXCL2, reverse EMT, and reduce the characteristic expression of CSC to inhibit the migration, invasion, and proliferation of 4T1 cells.
3.Research progress of new aluminum-containing vaccine adjuvants
Hongmei REN ; Yerong XIONG ; Xiaoying XUN ; Lei JIANG ; Jiasheng TU
Journal of China Pharmaceutical University 2025;56(2):236-243
Aluminum adjuvants are widely used in the field of vaccines due to their ability to induce efficient and long-lasting immune responses and good safety profile. With the development of immunology, the requirements for adjuvants have gradually increased, and traditional aluminum adjuvants can no longer meet all the needs of application. The development of novel aluminum adjuvants has become a hot research topic in order to achieve good immunity-enhancing effects and induce specific types and strengths of immune responses. This review briefly introduces the mechanism of action and safety of aluminum adjuvants, with focus on the research progress of novel aluminum adjuvants in recent years, mainly including nano-aluminum adjuvants and composite aluminum adjuvants (aluminum adjuvants compounded with immunity-stimulating molecules or delivery carriers), and a prospect of their future research direction, aiming to provide some reference for the further development and clinical application of aluminum adjuvants.
4.ZHANG Ren's academic characteristics of acupuncture for refractory eye diseases in modern times with "homotherapy for heteropathy".
Yue MA ; Yanmei HU ; Xiaolan SHI ; Xiaoying HU ; Wenqiang HONG ; Ren ZHANG
Chinese Acupuncture & Moxibustion 2025;45(9):1311-1317
This paper introduces the academic characteristics of Professor ZHANG Ren in treatment with acupuncture for refractory eye diseases in modern times, guided by "homotherapy for heteropathy" (same therapy for different diseases sharing the same pathogenesis). The refractory eye diseases in modern times include a variety of conditions such as glaucoma, macular degeneration, diabetic retinopathy, high myopia and its complications, dry eye, cortical visual impairment and genetic eye diseases. The same therapy is used because these diseases share the similar location and pathogenesis. Professor ZHANG optimizes the methods of acupoint selection and provides the comprehensive prescriptions, "basic prescription, prescription based on disease differentiation, and supplementary prescription". A variety of acupuncture manipulation techniques are operated in clinical practice, such as compound needling methods, penetration needling, manipulations for promoting qi movement and conducting qi flow. "Early, regular and persistent" treatment is the common requirement with "the same acupoints, the same prescription and the same acupuncture method" as well as at "the same time". It is also proposed that the treatment should be provided flexibly according to the different symptoms, "identifying the differences within similarities".
Acupuncture Therapy/methods*
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Humans
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Eye Diseases/history*
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Acupuncture Points
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History, 20th Century
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China
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History, 21st Century
5.Factors influencing of lymphopenia in prostate cancer patients during radiotherapy
Yifei LI ; Xianshu GAO ; Hongzhen LI ; Shangbin QIN ; Xin QI ; Mingwei MA ; Yun BAI ; Xueying REN ; Jiayan CHEN ; Feng LYU ; Xiaoying LI
Chinese Journal of Radiation Oncology 2025;34(4):347-354
Objective:To analyze the incidence and influencing factors of lymphopenia in prostate cancer patients undergoing pelvic radiotherapy.Methods:A retrospective analysis was conducted on 123 prostate cancer patients treated at the Department of Radiation Oncology, Peking University First Hospital, from November 2011 to May 2015. Radiotherapy was administered using conventional fractionated intensity-modulated radiotherapy. Blood routine, including absolute lymphocyte count (ALC), was performed on patients before radiotherapy, weekly during radiotherapy, and at the end of radiotherapy. Severe lymphopenia was defined as an ALC <500 cells/μl. Based on whether the minimum ALC during radiotherapy was lower than 500 cells/μl, the entire cohort and 55 patients (excluding those with undelineated pelvic bone marrow due to radiotherapy planning system issues) with delineated pelvic bone marrow (divided into pelvic bone marrow, iliac bone marrow, and lower pelvic bone marrow) were stratified into a severe lymphopenia group (33 cases and 16 cases, respectively) and a mild lymphopenia group (90 cases and 39 cases, respectively). Differences in clinical factors and dosimetric parameters were compared between the groups using the chi-square test (or Fisher's exact test), t-test, and Wilcoxon rank-sum test. Univariate and multivariate logistic regression analyses were performed to identify the clinical and dosimetric factors influencing severe lymphopenia. Results:All 123 prostate cancer patients experienced lymphopenia during radiotherapy, with a median minimum ALC of 0.6×10 9/L [range: (0.2-2.3)×10 9/L]. Severe lymphopenia occurred in 26.8% (33 cases) of patients. Univariate analysis of the entire cohort showed that pre-radiotherapy baseline ALC, initial neutrophil-to-lymphocyte ratio, prostate-specific antigen value, Gleason score, and pelvic radiotherapy were promoting factors for severe lymphopenia ( P<0.05). Multivariate analysis identified pre-radiotherapy baseline ALC ( OR=0.217, 95% CI: 0.072-0.650, P=0.006) and pelvic radiotherapy ( OR=23.852, 95% CI: 2.834-200.787, P=0.004) as promoting factors for severe lymphopenia. In patients with delineated pelvic bone marrow, univariate analysis showed that pelvic bone marrow V 30 Gy and V 40 Gy, iliac bone marrow V 30 Gy and V 40 Gy, lower pelvic bone marrow V 30 Gy and V 40 Gy were promoting factors for severe lymphopenia during treatment ( P<0.05). Conclusions:Lymphopenia is common in prostate cancer patients undergoing radiotherapy, with a high incidence of severe lymphopenia. Pre-radiotherapy baseline ALC, as well as pelvic, iliac, and lower pelvic bone marrow V 30 Gy and V 40 Gy, are promoting factors for severe lymphopenia during radiotherapy.
6.Expert consensus on holistic integrative management of oropharyngeal squamous cell carcinoma
Moyi SUN ; Zongxuan HE ; Qianwei NI ; Xiaoying LI ; Lin KONG ; Qing XI ; Wei GUO ; Zhangui TANG ; Guoxin REN ; Zhijun SUN ; Jian MENG ; Jie ZHANG ; Jichen LI ; Yue HE ; Chunjie LI ; Lizheng QIN ; Kai YANG ; Bing HAN ; Yan SUN ; Haijun LU ; Xiaohong ZHAN ; Dapeng HAO ; Kai SONG ; Haoyue XU ; Lingxue BU ; Jieying LI ; Man HU ; Mingjin XU ; Yun LI ; Wei SHANG
Journal of Practical Stomatology 2025;41(3):293-304
Oropharyngeal squamous cell carcinoma(OPSCC)is a malignant tumor originating from the squamous epithelium of the oro-pharyngeal mucosa,accounting for more than 90%of oropharyngeal malignancies.In recent years,human papillomavirus(HPV)infec-tion has become one of the primary etiological factors of oropharyngeal squamous carcinoma.The incidence of HPV-associated oropharyn-geal squamous carcinoma has been rising annually,with a noticeable trend toward younger populations,posing a significant threat to hu-man health.Due to the distinct biological behavior and clinical characteristics of HPV-associated oropharyngeal squamous carcinoma com-pared to its non-HPV-related counterpart,the diagnostic and treatment strategies for oropharyngeal squamous carcinoma have undergone substantial changes.Prevention and screening for oropharyngeal squamous carcinoma are of critical importance.The diagnostic and treat-ment process involves multi-disciplinary collaboration,including oral and maxillofacial surgery,otolaryngology,head and neck surgery,oncology,radiology and pathology.Based on evidence from clinical practice,a comprehensive,integrated diagnostic and therapeutic ap-proach has been established,centered around the concept of"prevention,screening,diagnosis,treatment,and rehabilitation",covering the entire patient lifecycle and providing a valuable reference for clinical practice.
7.Expert consensus on integrated diagnosis and treatment techniques for oropharyngeal squamous cell carcinoma
Wei SHANG ; Haoyue XU ; Zongxuan HE ; Xiaoying LI ; Haijun LU ; Xiaohong ZHAN ; Dapeng HAO ; Yan SUN ; Wei GUO ; Zhangui TANG ; Guoxin REN ; Zhijun SUN ; Jian MENG ; Jie ZHANG ; Jichen LI ; Yue HE ; Chunjie LI ; Jianhua WEI ; Lizheng QIN ; Yaowu YANG ; Qing XI ; Wei WU ; Kai YANG ; Bing HAN ; Lingxue BU ; Shuangyi WANG ; Kai SONG ; Jiaqi ZHU ; Hongyu HAN ; Yu KONG ; Jieying LI ; Man HU ; Mingjin XU ; Moyi SUN
Journal of Practical Stomatology 2025;41(6):725-736
In recent decades,the incidence of human papillomavirus(HPV)-associated oropharyngeal squamous cell carcinoma(OPSCC)has shown a marked increase.Significant changes have also occurred in the OPSCC diagnosis and treatment paradigm.Deter-mining HPV status prior to treatment is now essential,and radiotherapy/chemotherapy,immunotherapy,and minimally invasive surgical techniques have progressively emerged as key modalities for managing OPSCC.However,alongside these paradigm shifts,a comprehen-sive technical consensus guiding the entire diagnostic and therapeutic process for OPSCC patients is currently lacking.Given China's large population base and the rising incidence of OPSCC,an expert panel convened to develop a clinical technical consensus on OPSCC diagno-sis and management tailored to China's specific context.This consensus aims to further enhance and standardize understanding of OPSCC management techniques among relevant healthcare professionals.
8.The effect of hip-knee-ankle active and passive movement therapy on joint function in early and intermedi-ate-stage knee osteoarthritis patients
Xi LI ; Xiaoying REN ; Yongwei JIAO ; Zhipeng SUN ; Shilin YIN ; Zekun ZHANG ; Tianci GAO ; Jingxi WANG ; Yongwang ZHANG ; Lu LIU ; Shuangqing DU
The Journal of Practical Medicine 2025;41(6):829-837
Objective To evaluate the clinical efficacy of hip-knee-ankle active and passive exercise therapy in patients with early-to mid-stage knee osteoarthritis(KOA).Methods A total of 180 patients with early to mid-stage knee osteoarthritis(KOA)were recruited from the First Affiliated Hospital of Hebei University of Tradi-tional Chinese Medicine between March 2023 and March 2024.Patients were randomly assigned to one of four groups:active movement group,passive movement group,combined movement group,and control group,with 45 patients in each group.The active movement group received hip-knee-ankle active movement therapy daily until the end of follow-up.The passive movement group underwent hip-knee-ankle passive movement therapy three times per week for two weeks.The combined movement group received both active and passive therapies.The control group was administered oral celecoxib capsules(200 mg once daily for two weeks).Joint function was assessed in all four groups before treatment,at two weeks post-treatment,and at 14 weeks post-treatment.The primary outcome measure was the WOMAC joint function score,while secondary outcomes included the WOMAC pain score,stiffness score,and quality of life score(SF-12).Results A total of 160 patients completed the trial,with 39 in the active group,42 in the passive group,40 in the combined group,and 39 in the control group.There were no significant differences in baseline characteristics among the groups(P>0.05).Compared to baseline,the WOMAC scores for function,pain,and stiffness in the passive,combined,and control groups decreased significantly at both 2 and 14 weeks post-treatment(P<0.05),while the SF-12 scores increased significantly(P<0.05).Between 2 and 14 weeks post-treat-ment,the active and combined groups showed further significant decreases in WOMAC function,pain,and stiffness scores(P<0.05)and increases in SF-12 scores(P<0.05).At 2 weeks post-treatment,compared to the control group,the passive and combined groups exhibited significantly lower WOMAC function scores(P<0.05),with no significant difference between the passive and combined groups(P>0.05).By 14 weeks post-treatment,the active and combined groups demonstrated significantly lower WOMAC function scores(P<0.05),with the combined group showing a significantly lower score than the active group(P<0.05).Conclusion The four therapeutic approaches demonstrate a certain degree of efficacy in improving joint function for patients with early and mid-stage KOA.The passive therapy group exhibits superior short-term outcomes,while the active therapy group shows better long-term benefits.The combined therapy group presents notable advantages in both short-term and long-term effi-cacy,although its short-term effectiveness does not surpass that of the passive therapy group.It is recommended for patients with early and mid-stage KOA who have underlying gastrointestinal and cardiovascular conditions.
9.Expert consensus on holistic integrative management of oral squamous cell carcinoma
Moyi SUN ; Zongxuan HE ; Haoyue XU ; Xiaoying LI ; Jie ZHANG ; Haijun LU ; Xiaohong ZHAN ; Dapeng HAO ; Shizhu BAI ; Wei GUO ; Zhangui TANG ; Guoxin REN ; Jian MENG ; Zhijun SUN ; Jichen LI ; Yue HE ; Chunjie LI ; Lizheng QIN ; Kai YANG ; Qing XI ; Lin KONG ; Bing HAN ; Lingxue BU ; Yuanyong FENG ; Kai SONG ; Hongyu HAN ; Jieying LI ; Qianwei NI ; Yun LI ; Juan CHAI ; Xiaochen YANG ; Man HU ; Mingjin XU ; Wei SHANG
Journal of Practical Stomatology 2025;41(4):437-449
Oral squamous cell carcinoma(OSCC)is a malignant lesion originating from the oral mucosal squamous epithelium,account-ing for over 80%of oral and maxillofacial malignancies.Key etiological factors include tobacco,alcohol abuse,and betel quid chewing.In China,its incidence has shown an overall upward trend,posing a significant threat to public health.OSCC exhibits high local invasive-ness,making early diagnosis critical for improving prognosis.Its clinical management requires close multidisciplinary collaboration among oral and maxillofacial surgery,head and neck surgery,radiation oncology,medical oncology,reconstructive surgery,radiology,patholo-gy,and nutritional support teams.Given the increasing disease burden of OSCC and rapid development of multidisciplinary collaborative models,an expert panel has formulated this integrated management consensus based on evidence-based medicine and extensive deliber-ation.Centered on the'Prevention-Screening-Diagnosis-Treatment-Rehabilitation'framework,the consensus provides comprehensive guidance for the entire disease course of OSCC patients,aiming to standardize clinical practice.
10.Epidemiological characteristics of respiratory syncytial virus among inpatients in a children′s hospital in Shenzhen City from 2020 to 2023
Xiaojuan LUO ; Wei WANG ; Zhenmin REN ; Xiaoying FU ; Yunsheng CHEN ; Wenjian WANG ; Yanmin BAO ; Yuejie ZHENG ; Ke CAO ; Jiehua CHEN
Chinese Journal of Preventive Medicine 2025;59(4):484-489
This study analyzed the epidemiological characteristics and trends of respiratory syncytial virus (RSV) infections among inpatients with acute respiratory infections (ARI) in a children′s hospital in Shenzhen City inpatients from 2020 to 2023. From January 2020 to December 2023, multiple reverse transcription polymerase chain reaction (RT-PCR) combined with capillary electrophoresis fragment analysis technology was used to detect the nucleic acids of 12 respiratory pathogens, including RSV, in hospitalized children diagnosed with ARI. The patients were divided into six age groups: 0 to <6 months, 6 months to <1 year, 1 to <2 years, 2 to <5 years, 5 to <10 years, and 10 to <18 years. A total of 53 033 children were tested, including 6 830 RSV positive cases, with an overall positivity rate of 12.88%. The annual RSV positivity rates from 2020 to 2023 were 20.04%, 16.18%, 4.89%, and 13.33%, respectively, with statistically significant differences between the years ( χ2=1 185.994, P<0.001). The positive rate of RSV detection decreased with increasing age across all years (all P trend<0.05). From 2020 to 2023, the proportion of RSV-positive cases aged 2 to 5 years and older showed an increasing trend ( P trend<0.001 for all years). Compared to 2023, the median age of RSV-infected children was lower in 2020 ( Z=7.826, P<0.001) and 2021 ( Z=6.106, P<0.001). The proportion of severe infections requiring ICU admission did not change significantly across all years ( χ2=0.179, P=0.981). The RSV epidemic season in 2020 mainly occurred during 28-43 weeks, and in 2021, it spanned from 22-43 weeks. However, in 2022, the season was delayed until the 49th week and lasted for three weeks. In 2023, the seasonal epidemic appeared earlier, starting in the 14th week and lasting for 28 weeks. From 2020 to 2023, the rate of RSV co-infections with other pathogens (mycoplasma pneumoniae, human parainfluenza virus, human bocavirus, human coronavirus, human metapneumovirus, and influenza A) significantly increased (all P trend<0.01). In conclusion, the epidemiological characteristics of RSV infections in Shenzhen Children′s Hospital changed from 2020 to 2023. In 2022, there were only delayed, low-intensity and short-lived seasonal epidemics. However, in 2023, there was an earlier and prolonged epidemic, with increased infections in children aged 2 to 5 years and older and a rise in co-infections, while the proportion of severe infections requiring ICU admission remained unchanged.

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