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Objective:To observe the inhibitory effects of Huangqi Jiedu Decoction on lung metastasis of breast cancer in nude mice; To explore the mechanism of intervening epithelial mesenchymal transformation (EMT) induced by Wnt/β-catenin signaling pathway.Methods:Totally 30 nude mice were divided into model group, adriamycin group and Huangqi Jiedu Decoction low-, medium-, and high-dosage groups according to random number table method. Each group was injected subcutaneously with mouse breast cancer 4T1 cells to construct tumor - bearing nude mice model. Huangqi Jiedu Decoction low-, medium- and high-dosage groups were intragastrically administrated with Huangqi Jiedu Decoction 17.82, 35.64 and 71.28 g/kg; adriamycin group was injected intraperitoneally adriamycin 0.05 g/kg; model group was intragastrically administrated with normal saline of the same volume for 21 d. Tumor volume was measured at 9, 15, and 21 days after modeling. After the end of administration, the tumor tissue was separated, the tumor weight was measured, and the tumor inhibition rate was calculated. The lung tissue was Isolated,, the number of lung metastatic nodules and the inhibition rate of lung metastasis was counted. HE staining was used to observe the tissue morphology and evaluate the effectiveness of the model. The protein expressions of β-catenin, E-Cadherin and Vimentin in lung tissue were detected by Western Blot. The mRNA levels of β-catenin, E-Cadherin and Vimentin in lung tissue were detected by real-time fluorescent quantitative PCR.Results:Compared with the model group, the tumor volume and mass of Huangqi Jiedu Decoction low-, medium- and high-dosage groups decreased ( P<0.01); the number of pulmonary metastasis nodules in Huangqi Jiedu Decoction high-dosage group significantly decreased ( P<0.01); the mRNA and protein expressions of β-catenin and Vimentinm decreased in the Huangqi Jiedu Decoction low-, medium- and high-dosage groups ( P<0.01), and the protein and mRNA expressions of E-Cadherin increased in the Huangqi Jiedu Decoction high-dosage group ( P<0.01). Conclusion:Huangqi Jiedu Decoction can effectively inhibit the growth and lung metastasis of breast cancer transplanted tumor, and the mechanism may be to down-regulate the expression of key molecules in the Wnt/β-catanin signaling pathway, thereby inhibiting the EMT process, so as to inhibit the lung metastasis of breast cancer.
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Objective:To investigate the correlation between circadian blood pressure pattern and heart rate variability and stroke severity and outcome in patients with acute ischemic stroke (AIS).Methods:Patients with first-ever AIS admitted to the Affiliated Qingdao Central Hospital of Qingdao University from January 2015 to January 2021 were retrospectively included. Ambulatory blood pressure monitoring (ABPM) and ambulatory electrocardiogram (AECG) were performed after admission. The severity of stroke was assessed according to the National Institutes of Health Stroke Scale. ≤8 were defined as minor stroke, and >8 were defined as moderate to severe stroke. The modified Rankin Scale was used to evaluate the clinical outcome at 3 months after onset. ≤ 2 were defined as good outcomes, and >2 were defined as poor outcomes. Multivariate logistic regression analysis was used to determine the independent influencing factors of stroke severity and outcome. Results:A total of 516 patients with AIS were enrolled, including 328 male (63.57%), aged 59.62±6.67 years old. Among them, 266 patients (51.55%) were in the minor stroke group and 250 (48.45%) were in the moderate to severe stroke group. There were 463 patients (89.73%) were in the good outcome group and 53 (10.27%) were in the poor outcome group. Multivariate logistic regression analysis showed that hypertension (odds ratio [ OR] 5.021, 95% confidence interval [ CI] 2.635-10.923; P<0.001), atrial fibrillation ( OR 3.896, 95% CI 2.574-8.521; P<0.001), circadian blood pressure pattern (non-dipper type: OR 2.436, 95% CI 1.031-4.749, P<0.001; reverse dipper type: OR 2.654, 95% CI 1.642-5.268, P<0.001), SDNN ( OR 0.298, 95% CI 0.114-0.730; P=0.002), SDANN ( OR 0.325, 95% CI 0.200-0.679; P=0.009), rMSSD ( OR 0.437, 95% CI 0.255-0.876; P=0.016) and pNN50 ( OR 0.369, 95% CI 0.291-0.767; P=0.013) were the independent influencing factors of stroke severity. Hypertension ( OR 4.857, 95% CI 1.957-8.552; P<0.001), baseline NIHSS score ( OR 2.189, 95% CI 1.597-3.315; P<0.001), stroke severity ( OR 3.853, 95% CI 2.316-5.958; P<0.001), circadian blood pressure pattern (non-dipper type: OR 2.997, 95% CI 1.128-5.430, P<0.001; reverse dipper type: OR 3.703, 95% CI 1.478-5.902; P<0.001), SDNN ( OR0.369, 95% CI 0.215-0.779; P=0.015), SDANN ( OR 0.372, 95% CI 0.198-0.862; P=0.018), rMSSD ( OR 0.455, 95% CI 0.314-0.896; P=0.026) and pNN50 ( OR 0.448, 95% CI 0.307-0.825; P=0.021) were the independent influencing factors of poor outcomes. Conclusion:The non-dipper and reverse dipper circadian blood pressure patterns and lower heart rate variability are independently associated with stroke severity and poor outcomes in patients with AIS.
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ObjectiveTo investigate the mechanism of Chaihu Qinggantang （CHQGT） in the treatment of granulomatous lobular mastitis （GLM） in the rat model. MethodSixty female rats were divided into a normal group， a model group， a prednisolone group （0.001 8 g·kg-1）， and three CHQGT low-dose， medium-dose， and high-dose groups （4.5， 8.9， 17.8 g·kg-1）. The tissue homogenates mixed with GLM lesion tissue and Fritner's reagent were used for modeling. After modeling， the treatment groups were given corresponding treatment factors， and the normal group and the model group were given the equal volume of normal saline. The changes in mammary gland of rats were observed after 14 d. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in breast samples. The mRNA expressions of NOD-like receptor thermal protein domain associated protein 3 （NLRP3） inflammasome， Caspase-1， and interleukin-1β （IL-1β） were detected by real-time quantitative fluorescence polymerase chain reaction （Real-time PCR）. The protein expressions of NLRP3， Caspase-1， IL-1β， and IL-18 were detected by Western bolt. ResultAs compared with the normal group， the breasts of rats in the model group were obviously swelling， and mammary gland inflammation index was significantly increased （P<0.01）. Pathological changes included the formation of granuloma centered on the lobule of mammary gland with a large number of inflammatory cells such as lymphocytes and plasma cells. The mRNA expressions of NLRP3， Caspase-1， and IL-1β， and the protein expressions of NLRP3， Caspase-1， IL-1β， and IL18 in the model group were significantly increased （P<0.01）. Compared with the model group， the treatment groups improved breast swelling， and the CHQGT medium and high-dose groups and the prednisolone group reduced inflammation index to some extent after treatment （P<0.05， P<0.01）. The inflammation degree of mammary gland was significantly improved， and inflammatory cells such as macrophages， lymphocytes， and plasma cells were reduced to varying degrees in pathological aspects. The mRNA expressions of NLRP3， Caspase-1， and IL-1β， and the protein expressions of NLRP3， Caspase-1， IL-1β， and IL-18 in the CHQGT high-dose group and the prednisolone group were significantly down-regulated （P<0.05， P<0.01）. ConclusionCHQGT inhibits inflammation and treats GLM in rats. The mechanism is possibly related to the inhibition of NLRP3/IL-1β signaling pathway， which provides a new target for the prevention and treatment of GLM by Qingxiao method.
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Objective To observe the biological activity of the recombinant immunotoxin hIL-2-LuffinP1 derived by gene engineering in vitro.Methods To observe the inhibitory effect on lymphocytes in vitro,the splenocytes isolated from C57 and BALB/c mice were used for mixed mouse lymphocyte response(MLR),and the lymphocyte proliferation was observed in vitro with BALB/c mice splenocytes stimulated by concanavalin A(ConA).Different concentrations of hIL-2-LuffinP1 was added into the reaction system,with LuffinP1 adopted as control.3H-TdR incorporation assay was used to detect the effects of hIL-2-LuffinP1 on lymphocyte proliferation,and its effects on CTLL-2 cells(IL-2 dependent cells) were also observed.The experiments consisted of six groups(PBS,IL-2,LuffinP1,LuffinP1+IL-2,hIL-2-LuffinP1 and hIL-2-LuffinP1+IL-2).CTLL-2 cells were cultured for 12h after being treated with different agents,and then the cells were stained by trypan blue to estimate the dead cells in every 100 cells.Results It was showed that CPM declined correspondingly to the gradually increased concentration of hIL-2-LuffinP1,and cell proliferation was inhibited obviously.The inhibition rate was up to 97% at the concentration of 10-6mmol/L of hIL-2-LuffinP1.The activity of inhibiting lymphocyte proliferation was proportional to the dosage.On the contrary,no obvious inhibition to lymphocyte proliferation was found in the control group treated with LuffinP1,the inhibition ratio only reached to 14% with the same concentration of hIL-2-LuffinP1.It was also found that,when hIL-2-LuffinP1 and LuffinP1 in different concentrations added to spleen cells stimulated by ConA,hIL-2-LuffinP1 still exhibited strong ability of inhibition on the splenocyte proliferation,while LuffinP1 has no obvious inhibiting effect on the lymphocyte proliferation in the two reaction systems mentioned above.MLR showed the same results.Furthermore,in the experiment of cytotoxic effects of IL-2-LuffinP1 on CTLL-2 cells,the inhibition rates of hIL-2-LuffinP1+IL-2 group and hIL-2-LuffinP1 group were 29.6% and 47.4% respectively,the difference was significant compared with IL-2 group(P