Effect and mechanism of Qingre Huashi decoction on drug-resistant Helicobacter pylori

BACKGROUND Helicobacter pylori(HP),the most common pathogenic microorganism in stomach,can induce inflammatory reactions in the gastric mucosa,causing chronic gastritis and even gastric cancer.HP infection affects over 4.4 billion people globally,with a worldwide infection rate of up to 50%.The multidrug resistance of HP poses a serious challenge to eradication.It has been monstrated that compared to bismuth quadruple therapy,Qingre Huashi decoction(QHD)combined with triple therapy exhibits comparable eradication rates but with a lower incidence of adverse reactions;in addition,QHD directly inhibit and kill HP in vitro.METHODS In this study,12 HP strains were isolated in vitro after biopsy during gastroscopy of HP-infected patients.In vitro,the minimum inhibitory concentration(MIC)values for clinical HP strains and biofilm quantification were determined through the E-test method and crystal violet staining,respectively.The most robust biofilm-forming strain of HP was selected,and QHD was evaluated for its inhibitory and bactericidal effects on the strain with strong biofilm formation.This assessment was performed using agar dilution,E-test,killing dynamics,and transmission electron microscopy(TEM).The study also explored the impact of QHD on antibiotic resistance in these HP strains with strong biofilm formation.Crystalline violet method,scanning electron microscopy,laser confocal scanning microscopy,and(p)ppGpp chromatographic identification were employed to evaluate the effect of QHD on biofilm in strong biofilm-forming HP strains.The effect of QHD on biofilm and efflux pump-related gene expression was evaluated by quantitative polymerase chain reaction.Non-targeted metabolomics with UHPLC-MS/MS was used to identify potential metabolic pathways and biomarkers which were different between the NC and QHD groups.RESULTS HP could form biofilms of different degrees in vitro,and the intensity of formation was associated with the drug resistance of the strain.QHD had strong bacteriostatic and bactericidal effects on HP,with MICs of 32-64 mg/mL.QHD could inhibit the biofilm formation of the strong biofilm-forming HP strains,disrupt the biofilm structure,lower the accumulation of(p)ppGpp,decrease the expression of biofilm-related genes including LuxS,Spot,glup(HP1174),NapA,and CagE,and reduce the expression of efflux pump-related genes such as HP0605,HP0971,HP1327,and HP1489.Based on metabolomic analysis,QHD induced oxidative stress in HP,enhanced metabolism,and potentially inhibited relevant signaling pathways by upregulating adenosine monophosphate(AMP),thereby affecting HP growth,metabolism,and protein synthesis.CONCLUSION QHD exerts bacteriostatic and bactericidal effects on HP,and reduces HP drug resistance by inhibiting HP biofilm formation,destroying its biofilm structure,inhibiting the expression of biofilm-related genes and efflux pump-related genes,enhancing HP metabolism,and activating AMP in HP.

Effect of Qingre Huayu Decoction on Autophagy and Apoptosis of Neurons in Rats with Ischemic Brain Injury by Regulating Ca^(2+)/CaMKKβ-AMPK-mTOR Pathway

[Objectives]To explore the neuroprotective mechanism of Qingre Huayu Decoction on rats with acute cerebral ischemia injury.[Methods]SD rats were divided into sham operation group,ischemia model group,low,medium and high dose groups of Qingre Huayu De-coction,with 10 rats in each group.Referring to the MCAO operation model,both the sham operation group and the model group were given normal saline by gavage,and the Qingre Huayu Decoction group was given different doses of Qingre Huayu Decoction by gavage.After the op-eration,the rats were scored for neurological deficit,neurons were stained with HE,apoptotic cells were detected with TUNEL,and the levels of autophagy and apoptotic proteins in the Ca^(2+)/CaMKKβ-AMPK-mTORpathway in brain tissue were detected with Western-blot.[Results]Compared with the model group,the neurological function score of Qingre Huayu Decoction Group decreased significantly(P<0.05),the pathological damage of neurons in Qingre Huavu Decoction Group decreased.the proportion of apoptosis-positive cells detected by TUNEL de-creased(P<0.05),and the expression of CaMKKβand AMPK increased,expression of mTOR decreased,expression of Beclin-1 and LC3 increased,and expression of Caspase-3decreased in Qingre Huayu Decoction Group(P<0.05).[Conclusions]Qingre Huayu Decoction may play a neuroprotective role by activating Ca^(2+)/CaMKKβ-AMPK-mTOR pathway and regulating the level of apoptosis and autophagy.

Effect of Qingre Buyi Decoction (清热补益汤) on Nitric Oxide and Histomorphology of Intestinal Mucosa in Rats with Radiation Enteritis

Objective: To explore the mechanism of Qingre Buyi Decoction (清热补益汤,QBD) in prevention and treatment of radiation enteritis in rats. Methods: Forty-eight Wistar rats were randomly divided into four groups, the TCM group, the WM group, the model group and the control group, 12 in each group. Rats in the former three groups were given orally with QBD, norfloxacin and normal saline once a day for 7 successive days, after being irradiated with X-ray at single dose of 10 Gy for modeling of radiation enteritis, while rats in the control group were untreated. Animals were sacrificed at the end of the medication. NO concentration, mean height and number of villi per centimeter in their small intestinal mu-cosa were measured. Results: The intestinal NO concentration was significantly lower in the TCM and WM groups than that in the model group(P<0.05), while the number of villi was significantly more and the height higher in the former two groups than those in the model group (P<0.01 for both), but no significant difference was shown between the TCM group and the WM group. Conclusion: QBD could inhibit the production of NO, increase the number and height of intestinal villi in rats with radiation enteritis, suggesting that it could reduce the inflammatory reaction of intestinal mucosa to irradiation, protect muco-sa from radiation damage, and promote the regeneration of mucosa.

A study of ultrasonography for the treatment of Qingre Liangxue decoction on blood-heat psoriasis

Objective:The clinical effectiveness of blood-heat psoriasis treated by Qingre Liangxue decoction and ultrasound imaging changes of lesions was observed.Methods:Patients with blood-heat psoriasis were selected and randomly grouped into two groups:the treatment group taking the Qingre Liangxue decoction orally,the control group taking the Compound Glycyrrhizin Tablets orally,both groups applying topical emollient lotion for a 12-week treatment period.Ultrasound images of target lesions and healthy skin were collected at different treatment times.Results:The epidermal thickness thought,the dermal density incremented as well as the subepidermal low-echogenic band(SLEB)width narrowed in both groups during the treatment.Conclusion:Qingre Liangxue decoction has signified efficiency in the treatment of blood-heat psoriasis,and high-frequency ultrasound has the advantages of non-invasive,timely and objective in the evolution.

Clinical Evaluation of Yangyin Qingre Shengji Decoction(养阴清热生肌汤)in Preventing and Treating Acute Radiation Esophagitis

Objective:To explore the prevention and treatment effect of Yangyin Qingre Shengji Decoction(养阴清热生肌汤)on acute radiation esophagitis.Methods:From May 2020 to February 2021,60 patients with malignant tumor who underwent radiotherapy were selected as the research objects.They were divided into two groups by odd and even number drawing,30 cases in the control group and 30 cases in the observation group.The control group took mixed solution orally,while the observation group took Yangyin Qingre Shengji Decoction.The clinical effects,occurrence time,duration,grading of acute radiation esophagitis and TCM syndrome scores of dry mouth and constipation were compared between the two groups.Results:The effective rate of the observation group was significantly higher than that of the control group(P<0.05);Compared with the control group,the time of acute radiation esophagitis was prolonged and the duration of symptoms was shortened in the observation group(P<0.05);The incidence of above gradeⅡin the observation group was significantly lower than that in the control group,and the incidence of below gradeⅠwas significantly higher than that in the control group(P<0.05);There was no significant difference in the scores of dry mouth and constipation between the two groups before treatment(P>0.05),the scores of the two groups were decreased after treatment,and the observation group was lower than the control group(P<0.05).Conclusion:Yangyin Qingre Shengji Decoction(养阴清热生肌汤)has a positive effect on prevention and treatment of acute radiation esophagitis in patients with malignant tumor undergoing radiotherapy,which can be popularized.

A One-year Evaluation of Radiographic Progression in Patients with Rheumatoid Arthritis Treated by Qingre Huoxue Decoction(清热活血方)

Objective： TO investigate the effects of Qingre Huoxue Decoction （清热活血方, clearing heat and promoting blood flow; QRHXD）, on the radiographic progression in patients with rheumatoid arthritis （RA） by X-ray imaging. Methods： Eighty-six patients with active RA diagnosed as damp-heat and blood stasis syndrome were randomized into a QRHXD group and a QRHXD plus methotrexate （MTX） group, with 43 cases in each group. After one-year of treatment, 21 cases in each group （42 in total） were evaluated. Radiographs of hands were obtained at the baseline and after 12 months of treatment. Images were evaluated by investigators blinded to chronology and clinical data, and assessed according to the SharpNan der Heijde methods. Results： High intrareader agreements were reached （mean intraobserver intraclass coefficients： 0.95）. No significant change in any imaging parameters of joint destruction was observed at 12 months in either group; and the differences between the two groups were not significant （P〉0.05）. The mean of the changing score in the QRHXD group was 3.5 ± 4.1, and 2.4 ± 3.5 in the QRHXD＋MTX group, while the baseline radiographic score of patients in the QRHXD group was relatively higher （18.9± 19.1 vs. 14.0± 14.0）. The mean rates of the changing scores of the two groups were similar （0.24 ± 0.28 vs. 0.25± 0.44, P=0.40）. The severity of progression in the two groups was also similar （P=0.46）, 7 cases without radiographic progression in the QRHXD group and 8 in the QRHXD＋MTX group, 3 cases with obvious radiographic progression in the QRHXD group and 1 in the QRHXD＋MTX group. Conclusion： Radiographic progression of RA patients in both groups is similar, indicating that the QRHXD Decoction has a potential role in preventing bone destruction.

Efficacy of Qingre Huashi decoction(清热化湿方) on infection of Helicobacter pylori:inhibiting adhesion,antioxidant,and anti-inflammation

OBJECTIVE: To investigate the phytochemical profile of Qingre Huashi decoction( 清 热 化 湿 方, QHD) and evaluate the mechanisms rationalizing the use of QHD against Helicobacter pylori(H. pylori)-associated gastritis. METHODS: QHD is composed of 11 herbs, which was prepared by a fixed Pharmacy and concentrated into clear paste. High-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(HPLC-QTOF/MS) was used to detect the phytochemical profile of QHD. The toxicity of QHD against H. pylori and human gastric epithelial cells was evaluated by the toxicology test and cell counting kit-8 assay. The adhesion model was constructed by incubating H. pylori and gastric mucosal epithelial cells for 2 h. The urease assay was used to examine the antiadhesion effects of QHD, and gene expression of adhesins was evaluated by quantitative polymerase chain reaction. The antioxidant activity was assessed by DCFHDA labeling. To evaluate the anti-inflammatory effect, the levels of pro-inflammatory cytokines in the culture supernatant were detected by enzyme-linked immunosorbent assay. RESULTS: HPLC-QTOF/MS profiling indicated the presence of primary compounds 1-20 in QHD. Drug concentration was determined as 1, 2, and 5 mg/m L by the toxic concentration of QHD against H. pylori and human gastric epithelial cells. QHD prevented H. pylori adhesion to the human gastric epithelial cells and reduced levels of reactive oxygen species. QHD also reduced the level of interleukin-8 and other proinflammatory cytokines that were upregulated by H. pylori infection. CONCLUSION: QHD could inhibit H. pylori adhesion, and exert antioxidant and anti-inflammatory effects in vitro.

Study of Qingre Liyan Decoction(清热利咽汤)in Treating and Preventing Acute Radioactive Oral Mucositis

Objective： To study the effect of Qingre Liyan Decoction （清热利咽汤 QRLYD） in the prevention and treatment of acute radiative oral mucositis （AROM）, and to explore the mechanism of QRLYD by detecting epidermal growth factor （EGF） and T lymphocytes （CD3, CD4, and CD8）. Methods： Sixty patients conforming with the standard were randomly assigned to two groups, 30 patients in each group. Patients in the trial group were treated with QRLYD, and those in the control group were treated with Dobell＇s solution, both groups receiving conventional radiation treatment. The treatment course for both groups was 6 weeks on average. Blood routine test, CD3, CD4, and CD8 in the peripheral blood and EGF in the saliva were detected one day before and on the 14th and 28th day of radio-therapy. Results： Patients in the trial group were in good condition with normal spirits and intake of food and drinks. The incidence of AROM is lower and the effect in preventing AROM is higher in the trial group than those in the control group （P〈0.05）. The EGF in saliva, and CD4 and CD8 in the blood of patients in the trial group were higher than those in the control group （P〈0.05）. Conclusion： QRLYD can cure and prevent AROM. The mechanism may be related with its effects in enhancing body immunity and promoting salivary EGF.

Safety and Efficacy of Qingre Buyi Decoction(清热补益煎剂)in the Treatment of Acute Radiation Proctitis:A Prospective,Randomized and Controlled Trial

Objective:To investigate the efficiency,safety,and possible mechanisms of Qingre Buyi Decoction(清热补益煎剂,QBD)in the treatment of acute radiation proctitis(ARP).Methods:This study was a single center,prospective,single blind,randomized,and placebo-controlled clinical trial.A total of 60 patients with ARP was equally and randomly distributed into the control group(conventional treatment)and the combination group(conventional treatment plus QBD).The changes of main Chinese medicine clinical symptoms and signs,in...

Effect of Bushen Qingre Yuyin Decoction on Salivary Secretion, Spleen Index, Submandibular Gland Index, Submandibular Gland Histomorphology, and aqp5 Expression in the Nonobese Diabetic Mouse Model

Objective:The objective of this study was to observe the effect of Bushen Qingre Yuyin Decoction on the spleen index,submandibular gland index,salivary secretion,submandibular gland morphology,and aquaporin 5(AQP5)expression levels in the submandibular gland.Materials and Methods:Fifty 8-week-old nonobese diabetic(NOD)female mice were randomly divided into the following five groups:model;hydroxychloroquine;and high-,medium-,and low-dose Bushen Qingre Yuyin Decoction prescription.The blank group consisted of ten 8-week-old Balb/C rats.Bushen Yuyin Qingre Decoction was administered by gavage at 20,40,and 80 g/kg in the low-,medium-,and high-dose groups,respectively.The hydroxychloroquine group was administered hydroxychloroquine at a dose of 80 mg/kg by gavage.The normal and model groups were administered the same amount of saline.After the different administrations,the amount of salivary secretion in the mice was regularly observed.After 12 weeks,the mice were sacrificed,and their submandibular gland tissues were excised,stained with hematoxylin-eosin,and pathologically scored.AQP5 expression levels in the tissues were detected using real-time polymerase chain reaction.Results:The spleen index of mice in the high-dose group was higher than that in the model group(P<0.05),and the submandibular gland index of the hydroxychloroquine group and all the Bushen Qingre Yuyin Decoction prescription groups was higher than that in the model group(P<0.05).After 6 weeks,the salivary volume of the hydroxychloroquine group and all the Bushen Qingre Yuyin Decoction prescription groups was significantly higher than that of the model group(P<0.05);the amount of lymphocyte infiltration in mice among each administration group was significantly reduced(P<0.05);and the AQP5 expression levels in the submandibular glands of mice had significantly increased in the hydroxychloroquine and high-dose groups(P<0.05),with the high-dose group showing the best effect.Conclusion:Bushen Qingre Yuyin Decoction can effectively increase the spleen and submandibular gland indexes of NOD mice,increase the amount of salivary secretion,reduce lymphocyte infiltration in submandibular gland tissue,and significantly increase AQP5 expression levels,which may be positively correlated with the administered dose.

Clinical Observation of Bronchial AsthmaTreeted with Qingre Dingchuan Decoction（清热定喘汤）

Seventy patients of bronchial asthma ( 40 were adult and 30 child) were treated by QingreDingchuan decoction (QRDCD) , the erythrocytic superoxide dismutase (SOD) and blood lipid peroxide(LPO) levels were observed before and after treatment. Results showed that the patients , SOD was de-creased while their LPO increased signiticantly than that ot normal control. After treatment the LPO reducedsignificantly while the SOD elevated, the total effective rate ot adult group was 77. 5% and that of child groupwas 96 . 7 % .