Objective:To analyze locoregional recurrence (LRR) pattern of patients with pT 1-2N 1 breast cancer after modified radical mastectomy, with and without adjuvant radiotherapy (RT). Methods:A total of 5442 eligible patients with breast cancer from 12 Chinese centers were included. The LRR sites and the effect of RT at different sites on recurrence in patients with and without RT were analyzed. The Kaplan-Meier method was used to calculate the cumulative LRR rate, and the difference was compared by the log-rank test.Results:With a median follow-up time of 63.8 months for the entire cohort, 395 patients developed LRR. The chest wall and supraclavicular fossa were the most common LRR sites, regardless of RT or molecular subtypes. The 5-year chest wall recurrence rates for patients with and without chest wall irradiation were 2.5% and 3.8%( P=0.003); the 5-year supraclavicular lymph nodal recurrence rates for patients with and without supraclavicular fossa irradiation were 1.3% and 4.1%( P<0.001); the 5-year axillary recurrence-free rates for patients with and without axillary irradiation were 0.8% and 1.5%( HR=0.31, 95% CI: 0.04-2.23, P=0.219); and the 5-year internal mammary nodal recurrence-free rates for patients with and without internal mammary nodal irradiation were 0.8% and 1.5%( HR=0.45, 95% CI: 0.11-1.90, P=0.268). Conclusions:The chest wall and supraclavicular fossa are the most common LRR sites of patients with pT 1-2N 1 breast cancer after modified radical mastectomy, which is not affected by adjuvant RT or molecular subtypes. The chest wall and supraclavicular fossa irradiation significantly reduce the risk of recurrence in the corresponding area. However, axillary and internal mammary nodal irradiation has no impact on the risk of recurrence in the corresponding area.

Ulcerative colitis （UC） is a chronic intestinal disease with unknown etiology， with main symptoms of abdominal pain， diarrhea， mucus， pus， and blood in the stool. It can be accompanied by various complications and has a high risk of developing to colon cancer. In recent years， the incidence of UC and related colon cancer has been increasing， which seriously affects human health and quality of life. The operation， immunosuppressant， etc. are the main approaches in the modern clinical treatment of UC and related colon cancer， but these methods all have different toxic and side effects， and the therapeutic effect is not ideal. For many years， traditional Chinese medicine （TCM） has attracted much attention in the treatment of UC and related colon cancer due to its slightly toxic side effects and remarkable curative efficacy. Huangqintang， derived from the Shang Han Lun （伤寒论）， is composed of Scutellariae Radix， Paeoniae Radix Alba， Glycyrrhizae Radix et Rhizoma， and Jujubae Fructus with the functions of clearing heat， checking diarrhea， harmonizing the middle， and relieving pain， and has a significant effect on the treatment of UC. Huangqintang has complex compositions and plays roles with multiple targets and pathways. According to the literature and the research results of this research group for many years， it was found that the mechanism of Huangqintang in the treatment of UC and related colon cancer was presumably related to the protection of the intestinal mucosal barrier， inhibition of inflammatory response， promotion of mitophagy， inhibition of oxidative stress， regulation of intestinal flora， cell cycle， and gene expression， suppression of cell proliferation， and promotion of apoptosis. To provide theoretical references for an in-depth study of the mechanism and clinical use of Huangqintang， this paper reviewed the research advances in recent years.

ObjectiveTo evaluate the pharmacodynamic effect of Huangqintang （HQT） on ulcerative colitis （UC） model mice and investigate its protective effect against UC by regulating intestinal flora. MethodMale Balb/c mice were randomly divided into control group，model group， high-， medium-， and low-dose HQT groups （20， 10， 5 g·kg^-1）， flora interference group， flora interference model group， and flora interference-drug treatment group （HQT， 20 g·kg^-1）. The flora interference model was constructed through intragastric administration of antibiotics （200 mg·kg^-1 bacitracin and 200 mg·kg^-1 vancomycin） for 8 d， and the UC model was constructed by allowing mice with free access to 3% dextran sulfate sodium （DSS） solution for 7 d. HQT was administered for 7 d. After the experiments， the mice were sacrificed， and blood， colon， and feces were collected. Hematoxylin-eosin （HE） staining was performed to observe the colonic lesions. The serum levels of interleukin （IL）-4， IL-6， IL-10， and tumor necrosis factor （TNF）-α were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expression of Claudin1， MUC1， Occludin， and zonula occludens-1（ZO-1） in colon tissues was detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. The fecal DNA of mice was extracted and analyzed by high-throughput sequencing. ResultCompared with the normal group， the model group showed increased serum content of IL-4， IL-6， and TNF-α （P<0.05， P<0.01） and decreased IL-10 （P<0.05）. Compared with the model group， the HQT groups displayed decreased serum levels of IL-4， IL-6， and TNF-α （P<0.05， P<0.01）， increased IL-10 content （P<0.01）， increased mRNA and protein expression levels of Claudin1， MUC1， Occludin， and ZO-1 （P<0.05， P<0.01）. After flora interference， the diversity and abundance of intestinal bacteria decreased. To be specific， Proteobacteria increased （P<0.01）， and Firmicutes and Bacteroidetes decreased （P<0.01）. After UC induction by DSS， Bacteroidetes and Tenericutes decreased （P<0.05）. The high-， medium-， and low-dose HQT groups showed increased Bacteroidetes and Tenericutes （P<0.05， P<0.01） and decreased Firmicutes （P<0.05）. Additionally， the abundance of Lactobacillus， Lachnospiraceae NK4A136 group， Escherichia-Shigella， and Helicobacteris was positively proportional to the dose of HQT. ConclusionHQT can inhibit the inflammatory response of UC mice， restore the imbalance of intestinal flora， and repair the damaged intestinal mucosal barrier.

ObjectiveTo explore the anti-inflammatory mechanism of Huangqintang based on the inflammation model in RAW264.7 cells. MethodHuangqintang was prepared and the safe dose to RAW264.7 cells was screened out. The RAW264.7 cells were seeded in 24-well plates and incubated with Huangqintang and lipopolysaccharide （LPS）， successively. The concentrations of nitric oxide （NO）， interleukin （IL）-6， tumor necrosis factor （TNF）-α， and prostaglandin E₂ （PGE₂） were measured by Griess assay and enzyme-linked immunosorbent assay （ELISA）， respectively. Meanwhile， RAW264.7 cells were inoculated in 6-well plates， and normal group， LPS group， LPS+Huangqintang group， nuclear factor-κB （NF-κB） p65 inhibitor PDTC group， p38 mitogen-activated protein kinase （MAPK） inhibitor SB203580 group， extracellular signal-regulated kinase （ERK） inhibitor PD98059 group， c-Jun N-terminal kinase （JNK） inhibitor SP600125 group， and Janus kinase （JAK） inhibitor AG490 group were set up. After the cells were incubated with corresponding inhibitors and Huangqintang and stimulated by LPS， RNA and protein were extracted. The mRNA and protein expression levels of NF-κB p65， p38 MAPK， ERK， JNK， and JAK were detected by Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively， to explore the anti-inflammatory mechanism of Huangqintang by regulating the NF-κB， MAPK， and JAK/signal transducer and activator of transcription protein （STAT） signaling pathways. ResultAfter stimulation with LPS， the concentrations of NO， IL-6， TNF-α， and PGE₂ in the cells of the model group increased significantly（P<0.05，P<0.01）. Compare with the model group， after incubation with Huangqintang， the secretion of NO， IL-6， TNF-α， and PGE₂ showed a downward trend （P<0.05，P<0.01）. Compared with the normal group， the model group showed increased mRNA expression of p38 MAPK， ERK， JNK， JAK， and NF-κB p65 and total protein expression in cells after stimulation with LPS （P<0.05，P<0.01）. Compare with the model group，after incubation with Huangqintang， the total protein and mRNA expression of p38 MAPK， ERK， JNK， JAK， and NF-κB p65 in inflammatory cells decreased （P<0.05，P<0.01）. Meanwhile， the expression of NF-κB p65 total protein and mRNA in each inhibitor group showed a downward trend （P<0.05，P<0.01）. ConclusionHuangqintang can inhibit the inflammatory response through the NF-κB， MAPK， and JAK-STAT signaling pathways.

ObjectiveTo verify the anti-oxidative stress effect of Huangqintang based on the nuclear factor E₂-related factor 2 （Nrf2） signaling pathway by using Caco-2 cells as a carrier and RNA interference （RNAi） technology with in vitro experiments. MethodThe Caco-2 cells in the logarithmic growth phase were transfected with siRNA to construct siRNA Caco-2 cells. After normal Caco-2 cells and siRNA Caco-2 cells were incubated with Huangqintang of different doses， RNA and protein were extracted. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the mRNA and protein expression of heme oxygenase-1 （HO-1）， NAD（P）H quinone oxidoreductase 1 （NQO1）， glutathione S-transferase （GST）， Kelch-like ECH-associated protein 1 （Keap1）， and Nrf2. Meanwhile， the activities of superoxide dismutase （SOD） and GSH-Px， as well as the expression levels of malondialdehyde （MDA） and reactive oxygen species （ROS）， were detected by the colorimetric method and the probe method. ResultCompared with the results in the normal group， only the 400 mg·L^-1 Huangqintang group and the sulforaphane （SFN） group could reduce the content of ROS and MDA in Caco-2 cells （P<0.01）， while the activities of SOD and GSH-Px in the cells of the Huangqintang groups and the SFN group showed an upward trend. Furthermore， there were significant differences in the 400 mg·L^-1 Huangqintang group/the SFN group and the normal group （P<0.01）. Meanwhile， the protein and mRNA expression levels of HO-1， GST， Keap1， NQO1， and Nrf2 showed an upward trend in all groups （P<0.05， P<0.01）. After transfection， compared with the normal group， the model group showed increased content of MDA and ROS， blunted activities of GSH-Px and SOD， and reduced protein and mRNA expression of HO-1， GST， Keap1， and NQO1 （P<0.05， P<0.01）. After drug incubation， compared with the model group， the SFN group showed potentiated SOD activity， and the SFN group and the Huangqintang groups showed enhanced GSH-Px activity （P<0.01）. Moreover， the activities of SOD and GSH-Px in the 400 and 200 mg·L^-1 Huangqintang groups and the SFN group showed an upward trend （P<0.01）， and the content of MDA in the 400 mg·L^-1 Huangqintang group and the SFN group showed a downward trend. ROS decreased in all groups with drug intervention （P<0.01）， and the protein and mRNA expression of HO-1， GST， Keap1， NQO1， and Nrf2 increased to varying degrees （P<0.05， P<0.01）. ConclusionHuangqintang can play an anti-oxidative stress role by regulating the Nrf2 pathway.

ObjectiveTo study the effect of Huangqintang on intestinal T helper cells（Th）17/regulatory T cells（Treg） and Th1/Th2 balance in experimental ulcerative colitis （UC） mice. MethodSixty male C57BL/6J mice were randomly divided into normal group， model group， sulfasalazine group （0.2 g·kg^-1）， and Huangqintang low-dose， medium-dose and high-dose groups （4.55，9.1，18.2 g·kg^-1， respectively） according to body weight. Except for the normal group， all mice were given 2.5% dextran sodium sulfate （DSS， added to free drinking water） for 5 days to induce UC. After 7 days of continuous intragastric administration， the mice were sacrificed and the mesenteric lymph nodes （MLNs） and colon tissues were collected. The Th17/Treg and Th1/Th2 cell ratios in MLNs of mice were measured by flow cytometry. Interleukin-2 （IL-2）， interleukin-17A （IL-17A）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin-4 （IL-4）， interleukin-10 （IL-10） and other cytokines in colon tissues were detected by flow cytometric microsphere-based assay. The colon tissues samples were sectioned， and immunohistochemical method was used to detect the expression levels of T-box transcription factor （T-bet）， GATA binding protein 3 （GATA-3）， IL-17A and forkhead box P3 （FoxP3） in colon tissues. The content of transforming growth factor-β （TGF-β） in colon tissue of mice was determined by enzyme-linked immunosorbent assay （ELISA）. ResultCompared with the normal group， the number of Th1 and Th17 cells in the model group was significantly increased， while the number of Th2 and Treg cells was significantly decreased， and Th1/Th2 and Th17/Treg were significantly increased (P<0.05， P<0.01). The expressions of T-bet， GATA-3， IL-17A and FoxP3 in colon tissues of mice were significantly increased (P<0.05， P<0.01)， and the contents of cytokines IL-2， IL-17A， IL-6， TNF-α and IFN-γ were significantly increased. The contents of IL-4， TGF-β and IL-10 were significantly decreased (P<0.05， P<0.01). Compared with the model group， the imbalance of Th1/Th2 and Th17/Treg cells was reversed in each dose group of Huangqin decoction (P<0.01). The expressions of T-bet and GATA-3 in colon of mice were significantly decreased in low-dose， medium-dose and high-dose groups of Huangqintang (P<0.05， P<0.01)， and the expression of IL-17A was significantly decreased in high-dose and low-dose groups of Huangqintang (P<0.01). The contents of cytokines IL-2， IL-17A， IFN-γ， IL-6 and TNF-α were significantly decreased in the high-dose and medium-dose groups of Huangqintang， and the contents of TGF-β and IL-10 were significantly increased in the high-dose group of Huangqintang (P<0.05， P<0.01). ConclusionHuangqintang could reduce colonic inflammation and improve colonic tissue damage in UC mice， possibly by regulating Th17/Treg and Th1/Th2 balance and related cytokines levels， and the release level of pro-inflammatory cytokines.