1.Mechanism of Acupuncture at"Yanglingquan"and"Zusanli"in Regulating PINK1/Parkin-Mediated Mitophagy to Protect Chondrocytes in Rats with Knee Osteoarthritis
Weihua XUE ; Fangfang LUO ; Chuzhen LIN ; Limei XU
Journal of Guangzhou University of Traditional Chinese Medicine 2025;42(6):1446-1453
Objective To observe the therapeutic effects and mechanisms of acupuncture at"Yanglingquan"(GB34)and"Zusanli"(ST36)on knee osteoarthritis(KOA)rats,focusing on the regulation of PTEN-induced kinase 1/Parkin(PINK1/Parkin)-mediated mitophagy in chondrocytes.Methods Ten out of 40 male SD rats were randomly selected as the normal group,and the remaining 30 rats were used to construct a KOA model.A total of 27 rats were successfully modeled.The successfully modeled rats were randomly divided into a model group,an acupuncture group(acupuncture at Yanglingquan and Zusanli),and an acupuncture+autophagy inhibitor(3-MA)group(acupuncture at Yanglingquan and Zusanli combined with intraperitoneal injection of 3-MA),with 9 rats in each group.After intervention,the morphology of cartilage tissue in each group was observed,and the Mankin's score of cartilage was assessed.The apoptosis rate of chondrocytes was calculated.Enzyme-linked immunosorbent assay(ELISA)was used to detect inflammatory factors[cyclooxygenase 2(COX-2),interleukin 1β(IL-1β),and tumor necrosis factor α(TNF-α)],oxidative damage indicators[nitric oxide(NO),superoxide dismutase(SOD),malondialdehyde(MDA),and glutathione peroxidase(GSH-Px)],and the levels of matrix metalloproteinase 3(MMP-3)and tissue inhibitor of metalloproteinase 1(TIMP-1).The DHE probe was used to measure reactive oxygen species(ROS)level.Polymerase chain reaction(PCR)was used to detect the mRNA expression levels of PTEN-induced kinase 1(PINK1),Parkin,and microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)in cartilage tissue.Western blot was used to measure the protein expression levels of PINK1,Parkin,and LC3-Ⅱ in cartilage tissue.Results Compared with the normal group,the model group showed increased Mankin's score,apoptosis rate,COX-2 level,IL-1β and TNF-α levels,NO and MDA levels,MMP-3 and TIMP-1 levels,and ROS content,as well as decreased SOD and GSH-Px levels,and reduced mRNA and protein expressions of PINK1,Parkin and LC3-Ⅱ,with statistically significant differences(P<0.05).Compared with the model group,the acupuncture group showed decreased Mankin's score,apoptosis rate,COX-2 level,IL-1β and TNF-α levels,NO and MDA levels,MMP-3 and TIMP-1 levels,and ROS content,as well as increased SOD and GSH-Px levels,and elevated mRNA and protein expressions of PINK1,Parkin and LC3-Ⅱ,with statistically significant differences(P<0.05).Compared with the acupuncture group,the acupuncture+3-MA group showed increased Mankin's score,apoptosis rate,COX-2 level,IL-1β and TNF-α levels,NO and MDA levels,MMP-3 and TIMP-1 levels,and ROS content,as well as decreased SOD and GSH-Px levels,and mRNA and protein expressions of PINK1,Parkin and LC3-Ⅱ,with statistically significant differences(P<0.05).Conclusion Acupuncture at"Yanglingquan"and"Zusanli"can inhibit chondrocyte apoptosis by activating PINK1/Parkin-mediated mitophagy,thus improving the condition of KOA rats.
2.Clinical Experience of Qin Zhenhua in Treating Cutaneous Pruritus
Limei ZHU ; Yiban XU ; Junfeng TANG ; Jia LIN ; Wenliang ZHONG ; Zhenhua QIN
Journal of Guangzhou University of Traditional Chinese Medicine 2025;42(10):2578-2583
This study summarizes Professor Qin Zhenhua's clinical experience in treating cutaneous pruritus using a modified self-prescribed formula,Yinqiao Xingzi Decoction.Following the viewpoints from the classical Chinese medical literature and after years of clinical practice,Professor Qin proposes that the onset of cutaneous pruritus is often associated with the attack of exogenous pathogens,disharmony of qi and blood,or debility due to prolonged illness.The disease primarily involves the skin,with the fundamental pathogenesis of pathogens stagnating in the muscular superficies.Clinically,the treatment is frequently conducted from the perspective of fire-heat syndrome,and the therapeutic methods of clearing heat and removing toxins,releasing muscles and dispersing pathogens from the superficies,and dispelling wind to relieve itching are utilized.The approaches aim to cool heat in the blood,expel pathogens from the muscular layer,eliminate heat for stopping itching,and extinguish wind for calming the skin.The basic prescription,Yinqiao Xingzi Decoction(composed of Lonicerae Japonicae Flos,Forsythiae Fructus,Armeniacae Semen Amarum,Arnebiae Radix,Tribuli Fructus,Dictamni Cortex,Kochiae Fructus,and Glycyrrhizae Radix et Rhizoma),is used by modification depending on the predominance of pathogenic factors of wind,heat,dryness,or dampness-toxins,and according to the various syndromes like blood-heat,yin deficiency,or qi-blood deficiency.Yinqiao Xingzi Decoction is applicable to cutaneous pruritus with the most of clinical common syndrome types,but it is unsuitable for cases caused by yang deficiency or qi stagnation with blood stasis.
3.Application of 3D-HRAM model in diagnosis and localization of anal sphincter injury after surgery for complex anal fistula
Xuxiong WU ; Limei XU ; Juan HUANG ; Yaling YANG ; Zhaochu WANG ; Ruiwen LIANG ; Rong SHI
Journal of Chongqing Medical University 2025;50(6):821-827
Objective:To explore the application value of three-dimensional high-resolution anorectal manometry(3D-HRAM)model in the diagnosis and localization of anal sphincter injury after surgery for complex anal fistula.Methods:A prospective study was con-ducted on 92 patients who had undergone surgery for complex anal fistula in the Department of Coloproctology of The People's Hospi-tal Affiliated to Fujian University of Traditional Chinese Medicine from January 2023 to January 2024.The study was conducted in two stages.In the first stage,92 patients underwent 3D-HRAM and endoanal ultrasonography(EAUS)to examine the sphincter,and the data were compared with intraoperative records to evaluate the diagnostic efficiency and localization accuracy of 3D-HRAM and EAUS.In the second stage,92 patients who had been examined were grouped according to the presence or absence of symptoms of anal dysfunction to further verify the diagnostic and localization values of 3D-HRAM and EAUS for postoperative compensated/decompen-sated sphincter injury.Results:Among 92 patients,41 and 57 patients were diagnosed with sphincter injury through 3D-HRAM and EAUS,with a sensitivity of 61.19%and 85.07%,a specificity of 88%and 76%,an area under the curve of 0.739 and 0.813 calcu-lated by plotting the receiver operating characteristic curve,and a localization accuracy of 44.77%and 71.64%,respectively,showing significant differences between 3D-HRAM and EAUS(P=0.002),indicating that 3D-HRAM had lower diagnostic efficiency and local-ization accuracy than EAUS for anal sphincter injury after surgery for complex anal fistula.After grouping by symptoms,the diagnostic efficiency and localization accuracy of 3D-HRAM for symptomatic sphincter injury(decompensated)were significantly better than those for asymptomatic sphincter injury,and were similar to those of EAUS(P=0.763).Conclusion:The application advantage of 3D-HRAM lies in the diagnosis and localization of decompensated anal sphincter injury after surgery for complex anal fistula,which provides effective technical support for the study of anal defecation control mechanisms and injury compensation mechanisms.
4.Epidemiological characteristics of pulmonary tuberculosis among the elderly in Yangzhou City from 2013 to 2022
ZHAO Qianying ; WANG Hui ; LI Jincheng ; XU Jie ; ZHU Limei
Journal of Preventive Medicine 2025;37(3):276-279
Objective:
To investigate the epidemiological characteristics of pulmonary tuberculosis (PTB) among residents aged 60 years and older in Yangzhou City, Jiangsu Province from 2013 to 2022, so as to provide the evidence for the improvement of PTB prevention and control measures in this population.
Methods:
Data of PTB cases aged 60 years and older in Yangzhou City from 2013 to 2022 were collected from the Chinese Disease Prevention and Control Information System. The temporal, population, and regional distribution characteristics of PTB cases were analyzed using the descriptive epidemiological method.
Results:
A total of 8 726 PTB cases aged 60 years and older were registered in Yangzhou City from 2013 to 2022, including 4 167 cases positive for pathogenic tests, with a positive rate of 47.75%. The registered incidence rates and the positive rates of pathogenic tests of PTB among residents aged 60 years and older in Yangzhou City showed downward trends from 2013 to 2022 (both P<0.05). The average annual registered incidence rate of PTB was 83.95/105, and the average annual registered incidence rate of PTB positive for pathogenic tests was 40.09/105. The average annual registered incidence rate of PTB in males was higher than that in females (138.57/105 vs. 31.76/105, P<0.05). The registered incidence rate of PTB showed an increasing trend with age (P<0.05), with the highest rate observed in the age group of 75-<80 years (110.37/105). The top three regions with the highest average annual registered incidence rates of PTB were Jiangdu District (94.34/105), Baoying County (91.61/105) and Hanjiang District (84.93/105).
Conclusions
The registered incidence of PTB among residents aged 60 years and older in Yangzhou City showed a downward trend from 2013 to 2022. Particular attention should be payed to males, residents aged 75- <80 years, and the elderly in Jiangdu District and Baoying County.
5.Effect of serum HBV RNA on antiviral therapy in patients with chronic hepatitis B
Hongjing YIN ; Limei LI ; Xia CHENG ; Ying XU
Journal of Clinical Hepatology 2025;41(10):2030-2036
ObjectiveTo investigate the role of serum HBV RNA in assessing antiviral therapy for patients with chronic hepatitis B, as well as its potential as a biomarker in clinical therapy, and to provide a scientific basis for the clinical treatment of chronic hepatitis B. MethodsA total of 134 patients who were diagnosed with chronic HBV infection in The Second Affiliated Hospital of Kunming Medical University from April 2023 to May 2024 were enrolled as subjects, and related examinations were performed, including HBV DNA, serum HBV RNA, liver function, HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc, and transient elastography of the liver. The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the Kruskal-Wallis H test was used for comparison between multiple groups; the chi-square test or the continuity-adjusted chi-square test was used for comparison of categorical data between two groups. Pearson correlation analysis or Spearman correlation analysis was performed. ResultsThe 134 patients with chronic HBV infection were divided into HBeAg-positive group with 45 patients and HBeAg-negative group with 89 patients, and there were significant differences between the two groups in age, the positive rate and quantitative value of HBV DNA, the positive rate and quantitative value of serum HBV RNA, HBsAg, anti-HBe, and ALT (all P<0.05). In the cohort study of the HBeAg negative group, there were significant differences in the levels of HBV DNA, HBsAg, and GGT between the serum HBV RNA-negative group with 14 patients and the serum HBV RNA-positive group with 75 patients (all P<0.05). There were 28 patients in the HBeAg-positive group and 62 in the HBeAg-positive group who used antiviral drugs for ≥1 month, and the 28 HBeAg-positive patients had an HBV RNA positive rate of 100%, while for the 62 HBeAg-negative patients, there were significant differences in the positive rate and level of serum HBV RNA between the patients with different durations of medication (both P<0.05). Among the 89 HBeAg-negative patients, there were 62 treatment-experienced patients and 27 treatment-naïve patients, and there was a significant difference between the two groups in HBV RNA level [2.07 (1.52 — 2.82) log10 copies/mL vs 2.69 (1.80 — 3.55) log10 copies/mL, Z=2.034, P=0.042]. For HBeAg-negative patients, serum HBV RNA was positively correlated with HBV DNA and HBsAg (both P<0.05), and for HBeAg-positive patients, serum HBV RNA was significantly positively correlated with HBV DNA, HBsAg, and HBeAg (all P<0.05). ConclusionAntiviral therapy can reduce viral load, and for HBeAg-negative patients with high-sensitivity HBV DNA below the lower limit of detection, serum HBV RNA can fill the “gap” in the detection of viral replication.
6.A practice guideline for therapeutic drug monitoring of mycophenolic acid for solid organ transplants.
Shuang LIU ; Hongsheng CHEN ; Zaiwei SONG ; Qi GUO ; Xianglin ZHANG ; Bingyi SHI ; Suodi ZHAI ; Lingli ZHANG ; Liyan MIAO ; Liyan CUI ; Xiao CHEN ; Yalin DONG ; Weihong GE ; Xiaofei HOU ; Ling JIANG ; Long LIU ; Lihong LIU ; Maobai LIU ; Tao LIN ; Xiaoyang LU ; Lulin MA ; Changxi WANG ; Jianyong WU ; Wei WANG ; Zhuo WANG ; Ting XU ; Wujun XUE ; Bikui ZHANG ; Guanren ZHAO ; Jun ZHANG ; Limei ZHAO ; Qingchun ZHAO ; Xiaojian ZHANG ; Yi ZHANG ; Yu ZHANG ; Rongsheng ZHAO
Journal of Zhejiang University. Science. B 2025;26(9):897-914
Mycophenolic acid (MPA), the active moiety of both mycophenolate mofetil (MMF) and enteric-coated mycophenolate sodium (EC-MPS), serves as a primary immunosuppressant for maintaining solid organ transplants. Therapeutic drug monitoring (TDM) enhances treatment outcomes through tailored approaches. This study aimed to develop an evidence-based guideline for MPA TDM, facilitating its rational application in clinical settings. The guideline plan was drawn from the Institute of Medicine and World Health Organization (WHO) guidelines. Using the Delphi method, clinical questions and outcome indicators were generated. Systematic reviews, Grading of Recommendations Assessment, Development, and Evaluation (GRADE) evidence quality evaluations, expert opinions, and patient values guided evidence-based suggestions for the guideline. External reviews further refined the recommendations. The guideline for the TDM of MPA (IPGRP-2020CN099) consists of four sections and 16 recommendations encompassing target populations, monitoring strategies, dosage regimens, and influencing factors. High-risk populations, timing of TDM, area under the curve (AUC) versus trough concentration (C0), target concentration ranges, monitoring frequency, and analytical methods are addressed. Formulation-specific recommendations, initial dosage regimens, populations with unique considerations, pharmacokinetic-informed dosing, body weight factors, pharmacogenetics, and drug-drug interactions are covered. The evidence-based guideline offers a comprehensive recommendation for solid organ transplant recipients undergoing MPA therapy, promoting standardization of MPA TDM, and enhancing treatment efficacy and safety.
Mycophenolic Acid/administration & dosage*
;
Drug Monitoring/methods*
;
Humans
;
Organ Transplantation
;
Immunosuppressive Agents/administration & dosage*
;
Delphi Technique
7.Celastrol-loaded ginsenoside Rg3 liposomes boost immunotherapy by remodeling obesity-related immunosuppressive tumor microenvironment in melanoma.
Hongyan ZHANG ; Jingyi HUANG ; Yujie LI ; Wanyu JIN ; Jiale WEI ; Ninghui MA ; Limei SHEN ; Mancang GU ; Chaofeng MU ; Donghang XU ; Yang XIONG
Acta Pharmaceutica Sinica B 2025;15(5):2687-2702
Obesity usually exacerbates the immunosuppressive tumor microenvironment (ITME), hindering CD8+ T cell infiltration and function, which further represents a significant barrier to the efficacy of immunotherapy. Herein, a multifunctional liposomal system (CR-Lip) for encapsulating celastrol (CEL) was utilized to remodel obesity-related ITME and improve cancer immunotherapy, wherein Ginsenoside Rg3 (Rg3) was detected interspersed in the phospholipid bilayer and its glycosyl exposed on the surface of the liposome. CR-Lip had a relatively uniform size (116.5 nm), facilitating favorable tumor tissue accumulation through the interaction between Rg3 and glucose transporter 1 overexpressed in obese tumor cells. Upon reaching the tumor region, CR-Lip was found to induce the immunogenic cell death (ICD) of HFD tumor cells. Notably, the level of PHD3 in HFD tumor cells was effectively boosted by CR-Lip to effectively block metabolic reprogramming and increase the availability of major free fatty acids fuel sources. In vivo, experiments studies revealed that the easy-obtained nano platform stimulated enhanced the production of various cytokines in tumor tissues, DC maturation, CD8+ T-cell infiltration, and synergistic anticancer therapeutic potency with aPD-1 (tumor inhibition rate = 82.1%) towards obesity-related melanoma. Consequently, this study presented an efficacious approach to tumor immunotherapy in obese mice by encompassing tumor eradication, inducing ICD, and reprogramming metabolism. Furthermore, it offered a unique insight into a valuable attempt at the immunotherapy of obesity-associated related tumors.
8.Heart-sparing strategy for breast cancer radiotherapy based on nnU-Net: regional optimization and automatic segmentation
Jinghan HUANG ; Maidina BATUER ; Chuanghui ZHOU ; Zhi ZHANG ; Limei DENG ; Yuan XU ; Junyuan ZHONG ; Linghong ZHOU ; Xia LI ; Genggeng QIN
Chinese Journal of Radiation Oncology 2025;34(4):355-362
Objective:To investigate the feasibility and optimal expansion width of replacing the left anterior descending coronary artery (LADCA) with the region of heart sparing (RHS) to reduce cardiac radiation dose during breast cancer radiotherapy.Methods:Retrospective analysis was conducted on data from 88 patients with left-sided breast cancer who underwent radiotherapy at 2 centers: Nanfang Hospital of Southern Medical University (50 cases for the training set, 15 cases for the internal test set) and Ganzhou Hospital of Nanfang Hospital (23 cases for the external test set) from March 2022 to January 2024. All patients had left-sided invasive ductal carcinoma with axillary lymph node metastasis, and had undergone modified radical mastectomy and chemotherapy. Based on simulation CT images, 2 radiation oncologists delineated the LADCA and 8 RHSs. The RHSs were delineated by expanding the LADCA contour by 0.5 cm increments, totaling 8 expansions. The RHS widths were defined as 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, and 4.0 cm. The nnU-Net model was trained for 3D automatic segmentation of the LADCA and RHSs. Model performance was evaluated using the Dice similarity coefficient (DSC), relative volume error (RVE), sensitivity, specificity, and 95% Hausdorff distance (HD95). Additionally, the minimum, maximum, and average relative dose variations (RDV) as well as V5% and V20% indicators were calculated for the LADCA and each RHS. Correlation analysis was performed using the least squares regression, with the slope and coefficient of determination ( R2) employed to evaluate the accuracy of the model fitting, the relationship between the LADCA and RHS, and the degree of their correlation, thereby assessing the substitutive effect of the RHS for the LADCA. Results:The DSC for the LADCA was 0.415, while the DSCs for RHS widths of 0.5 cm and 4.0 cm were 0.718 and 0.835, respectively. Overall, the automatic segmentation performance improved with increasing RHS width. The DSC, RVE, sensitivity, specificity, and HD95 for the external test set were largely consistent with those of the internal test set, demonstrating the model's good robustness across different datasets. All RDVmin values were negative, while RDVmax and RDVmean showed a positive correlation with RHS width. RDVmean increased from 39.01% to 75.89% as the RHS width increased. In the correlation analysis, the slopes for RHS widths of 1.5 cm and 2.0 cm were 0.95 and 1.05, respectively, with R2 values and coefficients of variation of 0.79 and 0.73, and 21.11% and 24.03%, respectively. Conclusions:The automatic segmentation model trained on nnU-Net can accurately segment RHSs. Based on geometric and dosimetric indicators, a 1.5 cm-wide RHS is the most suitable substitute for the LADCA, effectively limiting the radiation dose to the LADCA without compromising target dose coverage.
9.Distribution of traditional Chinese medicine constitution and construction of a risk prediction model in patients with impaired awareness of hypoglycemia
Zhijia SHEN ; Qiaoyan LIU ; Zhijie QIAN ; Wentao SHI ; Limei YIN ; Lu XU
Chinese Journal of Practical Nursing 2025;41(15):1157-1167
Objective:To explore the distribution of Traditional Chinese Medicine constitution among patients with impaired awareness of hypoglycemia (IAH) and identify risk factors for IAH in patients with diabetes mellitus, to develop a risk prediction model. The aim is to validate the models′ predictive accuracy to facilitate early prevention and treatment of IAH.Methods:A case control study employing convenience sampling model was conducted on 1351 hospitalized patients with diabetes mellitus in the endocrinology departments of Changshu Hospital Affiliated to Nanjing University of Chinese Medicine and Affiliated Hospital of Jiangsu University, between August 2021 and December 2023. Traditional Chinese medicine constitution types were determined using the Traditional Chinese Medicine Constitution Classification and Judgment (ZYYXH/T157-2009). Data were divided into training and test sets at a ratio of 7∶3. Two prediction models were developed: Model 1, a conventional IAH prediction model for patients with diabetes mellitus, and Model 2, an IAH prediction model for patients with diabetes mellitus incorporating traditional Chinese medicine constitution. Nomograms were drawn for both models. The Hosmer-Lemeshow goodness-of-fit test, calibration curve, receiver operating characteristic (ROC) curve, and area under the curve (AUC) were calculated to evaluate the effectiveness of models 1 and 2. The improvement in prediction performance between Models 1 and 2 was assessed using Delong test, AUC, net reclassification improvement (NRI), integrated discrimination improvement (IDI), and decision curve analysis (DCA).Results:The study included 1 283 patients with diabetes mellitus, including 578 males and 705 females, aged (59.61 ± 14.09) years. The incidence of IAH among patients with diabetes mellitus was 20.50% (263/1283), with yang deficiency constitution being the most prevalent traditional Chinese medicine constitution type, at 47.53% (125/263). Multivariate analysis revealed that age, body mass index, course of diabetes, neurological hypoglycemia symptoms, hypoglycemia symptoms and severe hypoglycemia history were the influencing factors of Model 1 (all P<0.05); age, body mass index, neurological hypoglycemic symptoms, hypoglycemic symptoms, history of severe hypoglycemia, and traditional Chinese medicine constitution were the influencing factors of Model 2 (all P<0.05). The Hosmer-Lemeshow goodness-of-fit test showed a good fit of Model 2 [training set ( χ2=8.48, P>0.05), test set ( χ2=3.92, P>0.05)]. The Delong test results showed that the AUC for Model 2 was 0.96 for both the training and test sets, significantly higher than the AUCs of the 0.90 and 0.91 for Model 1 ( Z=-7.27, -3.70, both P<0.01). Furthermore, NRI was 0.66 ( 95%CI 0.53-0.79, P<0.01) and IDI was 0.02 (95% CI 0.01-0.03, P<0.05) for Model 2. Comparative analysis of clinical utility demonstrated that the net benefit of Model 2 for predicting IAH in patients with diabetes mellitus surpassed that of Model 1 across threshold probabilities ranging from 5% to 100%. Conclusions:The study constructed a nomogram prediction model included traditional Chinese medicine constitution with good predictive performance for IAH in patients with diabetes mellitus, and is of significant clinical value for identifying high-risk IAH populations.IAH patients mainly have a biased constitution, indicating that medical staff can reduce the incidence of IAH by improving the patients′ constitution.
10.Investigation and management of a clustered incident of incision infection after microtia plastic surgery
Jian SUN ; Zhiyuan CHEN ; Hua XU ; Limei MA ; Weiguang LI
Chinese Journal of Nosocomiology 2025;35(16):2432-2436
OBJECTIVE To summarize the process of investigation,disposal and prevention of a cluster incident of post-operative incision infection following microtia plastic surgery in the otorhinolaryngology department of a terti-ary care hospital,in order to provide a basis for hospital-aquired infection prevention and control.METHODS An epidemiological survey was conducted on seven patients who underwent otorhinolaryngoplasty for microtia in the ENT department of a tertiary care hospital from 27 Jul.to 7 Sep.2023,and interventions were implemented.RESULTS The incidence rate of infection was 71.43%among post-otorhinolaryngoplasty patients.In 5 patients,area for electrocoagulation of haemostatic increased by approximately 50%compared to the previous period,and drainage tubes were left in the surgical area for 11 to 13 days.Patients changed dressings in a shared dressing room.Pseudomonas aeruginosa was detected in three of the infected patients,and their antibiotic sensitivity pat-terns were similar to that of a lower respiratory tract infection patient(case 0)in the same department.CONCLUSION Inadequate aseptic practice by medical staff,longer retention of drainage tubes,incomplete disin-fection of the environment of the dressing room,and changes in surgical procedures may be the primary factors contributing to the occurrence of this cluster of infections.


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