1.Optimization of simmering technology of Rheum palmatum from Menghe Medical School and the changes of chemical components after processing
Jianglin XUE ; Yuxin LIU ; Pei ZHONG ; Chanming LIU ; Tulin LU ; Lin LI ; Xiaojing YAN ; Yueqin ZHU ; Feng HUA ; Wei HUANG
China Pharmacy 2025;36(1):44-50
OBJECTIVE To optimize the simmering technology of Rheum palmatum from Menghe Medical School and compare the difference of chemical components before and after processing. METHODS Using appearance score, the contents of gallic acid, 5-hydroxymethylfurfural (5-HMF), sennoside A+sennoside B, combined anthraquinone and free anthraquinone as indexes, analytic hierarchy process (AHP)-entropy weight method was used to calculate the comprehensive score of evaluation indicators; the orthogonal experiment was designed to optimize the processing technology of simmering R. palmatum with fire temperature, simmering time, paper layer number and paper wrapping time as factors; validation test was conducted. The changes in the contents of five anthraquinones (aloe-emodin, rhein, emodin, chrysophanol, physcion), five anthraquinone glycosides (barbaloin, rheinoside, rhubarb glycoside, emodin glycoside, and emodin methyl ether glycoside), two sennosides (sennoside A, sennoside B), gallic acid and 5-HMF were compared between simmered R. palmatum prepared by optimized technology and R. palmatum. RESULTS The optimal processing conditions of R. palmatum was as follows: each 80 g R. palmatum was wrapped with a layer of wet paper for 0.5 h, simmered on high heat for 20 min and then simmered at 140 ℃, the total simmering time was 2.5 h. The average comprehensive score of 3 validation tests was 94.10 (RSD<1.0%). After simmering, the contents of five anthraquinones and two sennosides were decreased significantly, while those of 5 free anthraquinones and gallic acid were increased to different extents; a new component 5-HMF was formed. CONCLUSIONS This study successfully optimizes the simmering technology of R. palmatum. There is a significant difference in the chemical components before and after processing, which can explain that simmering technology slows down the relase of R. palmatum and beneficiate it.
2.Chemical Constituents and Pharmacological Effect of Epimedium sagittatum: A Review
Lixin PEI ; Lin CHEN ; Nuo LI ; Mengyao ZHAO ; Haoyuan YANG ; Xiaoyu YANG ; Baoyu JI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):282-290
Epimedium sagittatum is a perennial herb of Berberidaceae. Its leaves have a long history of medicinal use in China. This plant is widely used as a Chinese traditional medicine,with the main functions of tonifying kidney Yang,strengthening bones and muscles,and dispelling wind and dampness. It can be used for treating kidney Yang deficiency,impotence,spermatorrhea,flaccidity of bones and muscles,rheumatic arthralgia,numbness,and spasms. The chemical constituents of this plant include flavonoids,polysaccharides,lignans,and alkaloids. Flavonoids are the main active ingredients. These compounds show a wide range of biological activities,including cartilage repair,anti-aging,anti-fatigue,cough-relieving,blood glucose-lowering,and anti-tumor effects. Modern pharmacological research has shown that E. sagittatum has definite pharmacological effects on the reproductive system,respiratory system,nervous system,cardiovascular system,skeletal system,etc. It has remarkable effects of helping pregnancy,resisting osteoporosis,controlling diabetes,improving immunity,and inhibiting tumor. Under the background of advocating one health and Chinese medicine,E. sagittatum is widely used in health care products,serving as the main raw material of various products. It has great market potential and is a Chinese medicinal herb with great clinical application and research value. This paper reviews the main chemical constituents and pharmacological effects of E. sagittatum based on domestic and foreign reports, providing a theoretical basis for further study on E. sagittatum and its safe clinical application.
3.Chemical Constituents and Pharmacological Effect of Epimedium sagittatum: A Review
Lixin PEI ; Lin CHEN ; Nuo LI ; Mengyao ZHAO ; Haoyuan YANG ; Xiaoyu YANG ; Baoyu JI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):282-290
Epimedium sagittatum is a perennial herb of Berberidaceae. Its leaves have a long history of medicinal use in China. This plant is widely used as a Chinese traditional medicine,with the main functions of tonifying kidney Yang,strengthening bones and muscles,and dispelling wind and dampness. It can be used for treating kidney Yang deficiency,impotence,spermatorrhea,flaccidity of bones and muscles,rheumatic arthralgia,numbness,and spasms. The chemical constituents of this plant include flavonoids,polysaccharides,lignans,and alkaloids. Flavonoids are the main active ingredients. These compounds show a wide range of biological activities,including cartilage repair,anti-aging,anti-fatigue,cough-relieving,blood glucose-lowering,and anti-tumor effects. Modern pharmacological research has shown that E. sagittatum has definite pharmacological effects on the reproductive system,respiratory system,nervous system,cardiovascular system,skeletal system,etc. It has remarkable effects of helping pregnancy,resisting osteoporosis,controlling diabetes,improving immunity,and inhibiting tumor. Under the background of advocating one health and Chinese medicine,E. sagittatum is widely used in health care products,serving as the main raw material of various products. It has great market potential and is a Chinese medicinal herb with great clinical application and research value. This paper reviews the main chemical constituents and pharmacological effects of E. sagittatum based on domestic and foreign reports, providing a theoretical basis for further study on E. sagittatum and its safe clinical application.
4.Clinical features of hepatitis B virus-related early-onset and late-onset liver cancer: A comparative analysis
Songlian LIU ; Bo LI ; Yaping WANG ; Aiqi LU ; Chujing LI ; Lihua LIN ; Qikai NING ; Ganqiu LIN ; Pei ZHOU ; Yujuan GUAN ; Jianping LI
Journal of Clinical Hepatology 2025;41(9):1837-1844
ObjectiveTo compare the clinical features of patients with hepatitis B virus (HBV)-related early-onset liver cancer and those with late-onset liver cancer, to assess the severity of the disease, and to provide a theoretical basis for the early diagnosis and treatment of liver cancer. MethodsA retrospective analysis was performed for 695 patients who were diagnosed with HBV-related liver cancer for the first time in Guangzhou Eighth People’s Hospital, Guangzhou Medical University, from January 2019 to August 2023, among whom 93 had early-onset liver cancer (defined as an age of50 years for female patients and40 years for male patients) and 602 had late-onset liver cancer (defined as an age of ≥50 years for female patients and ≥40 years for male patients). Related clinical data were collected, including demographic data, clinical symptoms at initial diagnosis, comorbidities, smoking history, drinking history, family history, routine blood test results, biochemical parameters of liver function, serum alpha-fetoprotein(AFP), virological indicators, coagulation function, and imaging findings. The pan-inflammatory indices neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR) were calculated, as well as FIB-4 index, aspartate aminotransferase-to-platelet ratio index (APRI), S index, Model for End-Stage Liver Disease (MELD) score, Child-Turcotte-Pugh (CTP) score, albumin-bilirubin (AIBL) grade, and Barcelona Clinic Liver Cancer (BCLC) stage. The independent-samples t test was used for comparison of normally distributed continuous data between two groups, and the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test or Fisher’s exact test were used for comparison of categorical data between two groups. ResultsThere were significant differences between the two groups in the proportion of male patients and the incidence rates of diabetes, hypertension, and fatty liver disease (χ2=6.357, 15.230, 11.467, and 14.204, all P0.05), and compared with the late-onset liver cancer group, the early-onset liver cancer group had a significantly higher proportion of patients progressing to liver cancer without underlying cirrhosis (χ2=24.657, P0.001) and a significantly higher proportion of patients with advanced BCLC stage (χ2=6.172, P=0.046). For the overall population, the most common clinical symptoms included abdominal distension, abdominal pain, poor appetite, weakness, a reduction in body weight, edema of both lower limbs, jaundice, yellow urine, and nausea, and 55 patients (7.9%) had no obvious symptoms at the time of diagnosis and were found to have liver cancer by routine reexamination, physical examination suggesting an increase in AFP, or radiological examination indicating hepatic space-occupying lesion; compared with the late-onset liver cancer group, the patients in the early-onset liver cancer group were more likely to have the symptoms of abdominal distension, abdominal pain, and jaundice (all P0.05). Compared with the late-onset liver cancer group, the early-onset liver cancer group had a significantly larger tumor diameter (Z=2.845, P=0.034), with higher prevalence rates of multiple tumors and intrahepatic, perihepatic, or distant metastasis (χ2=5.889 and 4.079, both P0.05), and there were significant differences between the two groups in tumor location and size (χ2=3.948 and 11.317, both P0.05). Compared with the late-onset liver cancer group, the early-onset liver cancer group had significantly lower FIB-4 index, proportion of patients with HBsAg ≤1 500 IU/mL, and levels of LMR and Cr (all P0.05), as well as significantly higher positive rate of HBeAg and levels of log10 HBV DNA, AFP, WBC, Hb, PLT, NLR, PLR, TBil, ALT, Alb, and TC (all P0.05). ConclusionCompared with late-onset liver cancer, patients with early-onset liver cancer tend to develop liver cancer without liver cirrhosis and have multiple tumors, obvious clinical symptoms, and advanced BCLC stage, which indicates a poor prognosis.
5.Effects of ginsenoside Rg3 on biobehavior of human gastric cancer SGC-7901 cells by regulating E2F1
Jian-Xin WANG ; Lin-Qian WANG ; Bo ZHU ; Pei SHI ; Yi-Chang SUN ; Li HAN
Chinese Pharmacological Bulletin 2024;40(5):853-858
Aim To explore the effect of ginsenoside Rg3 on the biological behavior of human gastric cancer SGC-7901 cells by regulating E2F1.Methods MTT assay was used to determine the effect of ginsenoside Rg3(0,80,160,320 μmol·L-1)on cell prolifera-tion.The effects of different concentrations of ginsen-oside Rg3 on apoptosis were measured by flow cytome-try.The effects of different concentrations of ginsen-oside Rg3 on cell migration and invasion were deter-mined by scratch healing experiment and Transwell ex-periment.The effects of different concentrations of gin-senoside Rg3 on the expression of E2F1,MMP-2,MMP-9,BCL-2 and Bax were determined by Western blot.Results Compared with the blank control group,the cell survival rate of 80,160 and 320 μmol ·L-1 ginsenoside Rg3 group was significantly lower,and it was concentration-dependent(P<0.05).Com-pared with the blank control group,the apoptosis rate of 80,160 and 320 μmol·L-1 ginsenoside Rg3 group significantly increased in a concentration-dependent manner(P<0.05).Compared with the blank control group,the number of cell migration in 80,160 and 320 μmol·L-1 ginsenoside Rg3 groups was significantly lower in a concentration-dependent manner(P<0.05).Compared with the blank control group,the number of cell invasion in 80,160 and 320 μmol· L-1 ginsenoside Rg3 groups was significantly lower in a concentration-dependent manner(P<0.05).The E2F1 mRNA and E2F1 protein expression in the 80,160,and 320 μmol·L-1 ginsenoside Rg3 groups were significantly reduced in a concentration-dependent manner compared with that in the blank control group(P<0.05).The protein expression of MMP-2,MMP-9,and BCL-2 in the cells of 80,160,and 320 μmol ·L-1 ginsenoside Rg3 group significantly decreased compared with those of the blank control group,and BCL-2 significantly increased compared with that of the blank control group in a concentration-dependent man-ner(P<0.05).Conclusions Ginsenoside Rg3 can reduce the proliferation,inhibit the migration and inva-sion of gastric cancer SGC-7901 cells,and promote the apoptosis of SGC-7901 cells in a concentration-depend-ent manner,and its mechanism may be related to the down-regulation of MMP-2,MMP-9,and BCL-2 ex-pression and up-regulation of Bax expression through E2F1.
6.Comparative study of total knee arthroplasty assisted by robot and remote sensing navigation system
Hai TANG ; Hong-Mei ZHANG ; Peng-Cheng SHAN ; Pei-Yan HU ; Lin JING ; Qi YAN ; Yuan-Yuan LI ; Xin-Yue WANG ; Si-Ye LIU ; Ming-Jiang HE
China Journal of Orthopaedics and Traumatology 2024;37(9):862-869
Objective To compare clinical efficacy of robot-assisted(RA)and remote sensing navigation alignment(RSNA)system-assisted total knee arthroplasty(TKA).Methods From March 2023 to June 2023,60 patients who underwent the first unilateral TKA due to severe knee osteoarthritis(KOA)were admitted and divided into RSNA group and RA group according to different treatment methods,with 30 patients in each group.There were 5 males and 25 females in RSNA group,aged from 56 to 81 years old with an average of(66.33±7.16)years old;body mass index(BM1)ranged from 19.87 to 38.54 kg·m-2 with an average of(28.40±6.18)kg·m-2;the courses of disease ranged from 5 to 36 months with an average of(18.20±8.98)months;RSNA system was used to assist the positioning of osteotomy.There were 7 males and 23 females in RA group,aged from 55 to 82 years old with an average of(67.83±8.61)years old;BMI ranged from 19.67 to 37.25 kg·m-2 with an aver-age of(28.01±4.89)kg·m-2;the courses of disease ranged from 3 to 33 months with an average of(17.93±9.20)months;RA was performed.Operation time,incision length,latent blood loss at 2 weeks after operation and incidence of lower extremity thrombosis were compared between two groups.Hip-knee ankle angle(HKAA),HKAA deviation,lateral distal femoral angle(LDFA),medial proximal tibial angle(MPTA)and posterior tibial slope(PTS)were compared between two groups;Western Ontario McMaster Universities Osteoarthritis Index(WOMAC)and Knee Society score(KSS)were used to evaluate functional recovery before operation,3 and 6 months after operation.Results The operation was performed successfully in both groups,and there were no serious complications such as vascular and nerve injury during operation.The wound healed well at stage Ⅰafter operation,and the follow-up time was 6 months.The operation time,latent blood loss at 2 weeks after operation and inci-sion length in RSNA group were(94.35±5.75)min,(130.54±17.53)mland(14.73±2.14)cm,respectively;while(102.57±6.88)min,(146.33±19.47)ml and(16.78±2.32)cm in RA group,respectively.RSNA group was better than RA group(P<0.05).No deep vein thrombosis occurred in both groups at 2 weeks after operation,5 patients occurred intermuscular vein throm-bosisin in RSNA group and 8 patients in RA group,the difference was not statistically significant(P>0.05).In RSNA group,HKAA,LDFA and MPTA were(173.00±5.54)°,(86.96±3.45)°,(82.79±3.35)° before operation,and(178.34±1.85)°,(89.92±0.42)°,(89.84±0.73)° at 1 week after operation,respectively.In RA group,HKAA,LDFA and MPTA were(173.31±6.48)°,(87.15±3.40)° and(82.99±3.05)° before operation,and(178.52±1.79)°,(90.03±0.39)° and(90.15±0.47)° at 1 week after operation,respectively.HKAA,LDFA and MPTA were significantly improved in both groups at 1 week after oper-ation(P<0.05).There were no significant difference in HKAA,LDFA,MPTA and PTS between two groups before operation and 1 week after operation(P>0.05).There was no significant difference in deviation distribution of HKAA at 1 week after op-eration(x2=2.61 1,P=0.456).There were no significant difference in WOMAC and KSS between two groups before operation,3 and 6 months after operation(P>0.05),and postoperative WOMAC and KSS at 3 and 6 months between two groups were im-proved compared with those before operation(P<0.05).Conclusion Both RA and RSNA system assisted TKA could obtain ac-curate osteotomy,RA has higher surgical accuracy,RSNA system assisted operation has less trauma,and operation is simpler.
7.Application of remote sensing navigation system in total knee arthroplasty
Yuan-Yuan LI ; Ming-Jiang HE ; Peng-Cheng SHAN ; Pei-Yan HU ; Lin JING ; Qi YAN ; Hai TANG ; Xin-Yue WANG ; Si-Ye LIU ; Hong-Mei ZHANG
China Journal of Orthopaedics and Traumatology 2024;37(9):878-885
Objective To explore clinical accuracy of remote sensing navigation alignment(RSNA)system in total knee arthroplasty(TKA)and its influence on postoperative clinical efficacy.Methods From May 2021 to May 2022,60 knee os-teoarthritis(KOA)patients with Kellgren-Lawrence(K-L)grade Ⅲ to Ⅳ treated by unilateral primary TKA were selected and divided into RSNA group and traditional operation group according to treatment methods,and 30 patients in each group.There were 6 males and 24 females in RSNA group,aged from 55 to 86 years old with an average of(68.06±8.23)years old;body mass index(BMI)ranged from 22.15 to 34.58 kg·m-2 with an average of(28.20±3.01)kg·m-2;the courses of disease ranged from 2 to 60 months with an average of(18.80±14.80)months;13 patients with grade Ⅲ and 17 patients with grade Ⅳaccording to K-L grading.In traditional operation group,there were 8 males and 22 females,aged from 57 to 85 years old with an average of(67.26±6.32)years old;BMI ranged from 23.94 to 34.55 kg·m-2 with an average of(27.49±2.32)kg·m-2;the courses of disease ranged from 3 to 60 months with an average of(21.30±16.44)months;14 patients with grade Ⅲ and 16 pa-tients with grade Ⅳ according to K-L grading.Western Ontario and McMaster Universities(WOMAC)osteoarthritis index and Knee Society score(KSS)were used to evaluate functional recovery of patients.Hip-knee-ankle angle(HKAA),distal femoral valgus angle(FVA)and distal fermoral flexion angle(DFFA)were measured before operation.HKAA and HKAA deviation angle were measured at 1 week after operation,and defective rate of lower limb force line,femur prosthesis valgus angle(FP-VA)and femoral prosthesis flexion angle(FPFA),respectively,were calculated.Results There were no serious complications such as vascular and nerve injury during operation,and wound healed at stage Ⅰ.Both groups were followed up for 6 months.There were no significant difference in WOMAC index,KSS,HKAA,FVA and DFFA between two groups before operation(P>0.05).The force line defect rate,HKAA,HKAA deviation angle,FPVA deviation angle and FPFA of RSNA group were 6.7%,(178.74±1.56)°,(1.25±1.56)°,(1.84±0.16)° and(4.85±2.46)°,respectively;while in traditional operation group were 20%,(176.73±3.46)°,(3.27±3.46)°,(2.44±0.26)°,(6.60±1.86)°;the difference between two groups were statistically sig-nificant(P<0.05).There were no significant difference in WOMAC index and KSS between two groups at 3 and 6 months after operation(P>0.05).Conclusion RSNA system could reduce defective rate of lower limb force line,FPVA deviation angle and FPFA after TKA,which is more accurate and easy to operate than traditional intramedullary localization surgery while ensuring postoperative efficacy.
8.Scoping review of self-advocacy needs and behaviors of adult cancer patients
Yuanyuan LI ; Lin CHENG ; Yulu XU ; Bei PEI ; Huan LI ; Jinlong LIU ; Yan LOU
Chinese Journal of Modern Nursing 2024;30(21):2916-2923
Objective:To conduct a scoping review of research on the self-advocacy of adult cancer patients to identify their self-advocacy needs, behaviors, strengths and weaknesses, so as to provide guidance for future research in this field.Methods:Using the scoping review guidelines of Joanna Briggs Institute in Australia as a methodological framework, relevant literatures were searched in PubMed, Web of Science, Cochrane Library, Embase, Science Direct, CINAHL, Scopus, China National Knowledge Infrastructure, Wanfang, VIP and China Biology Medicine disc. The search period was from establishment of the databases to December 2023, and the results were summarized and analyzed.Results:A total of 14 articles were included, and self-advocacy needs included six types of needs, such as symptom management, communication, interpersonal support, disease information, decision-making and emotional management. Self-advocacy included four aspects, including seeking information, self-decision-making, strengthening contact with the outside world and effective communication.Conclusions:Adult cancer patients have diverse types of self-advocacy needs and certain self-advocacy behaviors. Future research should pay attention to self-advocacy needs assessment, develop corresponding tools, focus on self-advocacy behaviors, leverage the advantages of self-advocacy and explore strategies to achieve effective self-advocacy support.
9.Application of mobile health applications in colorectal cancer patients: a scoping review
Bei PEI ; Yuanyuan LI ; Lin CHENG ; Meirong HONG ; Wanying WU ; Yan LOU
Chinese Journal of Modern Nursing 2024;30(33):4603-4610
Objective:To conduct a scoping review of the application of mobile health applications (MHA) in the care of patients with colorectal cancer, summarizing the development process, the functions achieved, as well as the evaluation metrics, to provide references for MHA practice and related research.Methods:Following the scoping review framework, a comprehensive search was conducted across both domestic and international databases, including the China Biomedical Literature Database, Wanfang Database, China National Knowledge Infrastructure (CNKI), VIP Database, Cochrane Library, PubMed, Embase, CINAHL, Web of Science, and Scopus. The search period was from the database inception to February, 2024.Results:A total of 16 studies were included. The development of MHA involved multiple methods including literature reviews, qualitative interviews, consultations with multidisciplinary teams, and guidance from theoretical models. The functions of MHA include health education, peer support, guided feedback, monitoring, and reminder features. Evaluation metrics for MHA comprise usability, adherence, and effectiveness.Conclusions:MHA has demonstrated positive effects in enhancing patients' knowledge and alleviating symptoms such as fatigue and vomiting in colorectal cancer patients. However, it is still in its early stages, and further high-quality studies are needed to scientifically develop MHA that meets patients' needs.
10.Establishment and Evaluation Strategy of an in Vitro Cell Model of Bone Marrow Microenvironment Injury in Mouse Acute Graft-Versus-Host Disease
Jia-Yi TIAN ; Pei-Lin LI ; Jie TANG ; Run-Xiang XU ; Bo-Feng YIN ; Fei-Yan WANG ; Xiao-Tong LI ; Hong-Mei NING ; Heng ZHU ; Li DING
Journal of Experimental Hematology 2024;32(2):617-624
Objective:To establish a mesenchymal stem cell(MSC)-based in vitro cell model for the evaluation of mouse bone marrow acute graft-versus-host disease(aGVHD).Methods:Female C57BL/6N mice aged 6-8 weeks were used as bone marrow and lymphocyte donors,and female BALB/c mice aged 6-8 weeks were used as aGVHD recipients.The recipient mouse received a lethal dose(8.0 Gy,72.76 cGy/min)of total body γ irradiation,and injected with donor mouse derived bone marrow cells(1× 107/mouse)in 6-8 hours post irradiation to establish a bone marrow transplantation(BMT)mouse model(n=20).In addition,the recipient mice received a lethal dose(8.0 Gy,72.76 cGy/min)of total body γ irradiation,and injected with donor mouse derived bone marrow cells(1 × 107/mouse)and spleen lymphocytes(2 × 106/mouse)in 6-8 hours post irradiation to establish a mouse aGVHD model(n=20).On the day 7 after modeling,the recipient mice were anesthetized and the blood was harvested post eyeball enucleation.The serum was collected by centrifugation.Mouse MSCs were isolated and cultured with the addition of 2%,5%,and 10%recipient serum from BMT group or aGVHD group respectively.The colony-forming unit-fibroblast(CFU-F)experiment was performed to evaluate the potential effects of serums on the self-renewal ability of MSC.The expression of CD29 and CD105 of MSC was evaluated by immunofluorescence staining.In addition,the expression of self-renewal-related genes including Oct-4,Sox-2,and Nanog in MSC was detected by real-time fluorescence quantitative PCR(RT-qPCR).Results:We successfully established an in vitro cell model that could mimic the bone marrow microenvironment damage of the mouse with aGVHD.CFU-F assay showed that,on day 7 after the culture,compared with the BMT group,MSC colony formation ability of aGVHD serum concentrations groups of 2%and 5%was significantly reduced(P<0.05);after the culture,at day 14,compared with the BMT group,MSC colony formation ability in different aGVHD serum concentration was significantly reduced(P<0.05).The immunofluorescence staining showed that,compared with the BMT group,the proportion of MSC surface molecules CD29+and CD 105+cells was significantly dereased in the aGVHD serum concentration group(P<0.05),the most significant difference was at a serum concentration of 10%(P<0.001,P<0.01).The results of RT-qPCR detection showed that the expression of the MSC self-renewal-related genes Oct-4,Sox-2,and Nanog was decreased,the most significant difference was observed at an aGVHD serum concentration of 10%(P<0.01,P<0.001,P<0.001).Conclusion:By co-culturing different concentrations of mouse aGVHD serum and mouse MSC,we found that the addition of mouse aGVHD serum at different concentrations impaired the MSC self-renewal ability,which providing a new tool for the field of aGVHD bone marrow microenvironment damage.

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