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@#Objective To detect the virulence gene of E. coli from calves with diarrhea in Tongliao City,Inner Mongolia Autonomous Region,China and analyze its antimicrobial resistance as well as the distribution of antimicrobial resistant genes. Methods The sensitivities of 82 E. coli isolates from the fecal samples of calves with diarrhea to thirteen kinds of antibiotics were determined by disk diffusion test. The carrying statuses of thirteen virulence genes and twelve antimicrobial resistant genes of the E. coli isolates were determined by PCR,based on which the phylogenetic background was investigated. Results Of the 82 pathogenic E. coli isolates,48. 78%(40/82)、31. 71%(26/82)、14. 63%(12/82)and 4. 88%(4/82)belonged to phylogenic groups A,B1,B2 and D respectively,indicating that the prominent one was group A. A total of 11 virulence genes were detected in 82 isolates. The detection rates of irp2,fyuA,eaeA and STb genes were 79. 27%(65/82),63. 41%(52/82),53. 66%(44/82)and 50%(41/82)respectively,while those of other virulence genes were less than 50%,and no tsh or LT1 was detected. The 82 isolates were significantly resistant to 13 kinds of antibiotics,in which the resistant rates to tetracycline,doxycycline and amoxicillin were 100%(82/82),97. 56%(80/82)and 90. 24%(74/82)respectively. All the isolates were mutidrug resistant,most of which were resistant to eight kinds of antibiotics(16/82,19. 51%). A total of twelve antimicrobial resistant genes were detected in the 82 isolates,in which the positive rates of genes resistant to β ‑ lactams(blaTEM),sulfonamide(sul1 and sul2),tetracycline(tetB and tetD)and aminoglycosides(aadB)were more than 70%. Conclusion The 82 pathogenic E. coli isolates mainly belonged to group A,with high detection rates of virulence gene and antimicrobial resistant gene as well as high and multiple drug resistance. The study provided a reference for the prevention and treatment of and clinical medication of E. coli‑associated diseases in calves in Tongliao Region.
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Teprotumumab-trbw,a monoclonal antibody that acts on the insulin growth factor-Ⅰ receptor, was approved in 2020 for the treatment of thyroid-associated ophthalmopathy, but little is known about it in China. It is hoped to provide guidance for clinical use through the review of its molecular structure, pharmacokinetics, therapeutic mechanism, clinical research and safety. It inhibits immune inflammation by blocking thyroid-stimulating hormone receptor /insulin growth factor-Ⅰ receptor crosstalk signaling, so as to reduce the production of hyaluronic acid and inflammatory factors in response. It can also promote the apoptosis of retro-orbital fibroblasts/adipocytes and inhibit the expression of genes related to the synthesis of thyroid hormones, thereby significantly improving the clinical symptoms such as exophthalmos and diplopia. The common adverse reactions of Teprotumumab-trbw are muscle spasm, hyperglycemia, hearing loss and so on. Teprotumumab-trbw is effective and durable in the treatment of thyroid-associated ophthalmopathy, and patients with secondary treatment can also benefit from it, which provides a new way and hope for the treatment of thyroid-associated ophthalmopathy.
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Objective:To investigate the predictive role of dynamic changes of plasma biomarkers in patients with viral and mycoplasma community-acquired pneumonia (CAP).Methods:From January 2020 to June 2020, 141 patients with viral and mycoplasma CAP in People's Hospital of Ningxia Hui Autonomous Region were enrolled. Pneumonia severity index (PSI) scores [grade Ⅰ-Ⅱ(PSI score ≤ 70), grade Ⅲ (PSI score 71-90) and grade Ⅳ-Ⅴ(PSI score ≥ 91)], serum amyloid A (SAA), hypersensitive C-reactive protein (hs-CRP), procalcitonin (PCT), erythrocyte sedimentation rate (ESR) and white blood cell (WBC) on the 1 day after admission were compared between the different pathogens (viral and mycoplasma) or different disease severity. The change in level of SAA, hs-CRP on the third day (Δ 3 d = 1 d-3 d) were compared among different disease outcome groups (patients were divided into improved group, stable group and exacerbation group based on PSI scores or lung CT images on the third day). The change in the level of SAA, hs-CRP on the seventh day (Δ 7 d = 1 d-7 d) were compared among different disease prognosis groups (patients were divided into survival group and death group based on 28-day survival data). The receiver operating characteristic curve (ROC) were drawn to evaluate the value of SAA in the evaluation of disease and prediction prognosis. Results:The level of SAA in mycoplasma group (43 cases) was significantly higher than that in virus group (98 cases) on the 1 day after admission. There were no significant differences in other plasma biomarkers between the two groups. The more severe the illness, the higher the SAA level on the 1 day after admission. The trends of other plasma biomarkers in the two groups were consistent with SAA. The levels of SAA in the patients with exacerbation of the virus group and mycoplasma group (12 cases, 9 cases) were significantly higher than those of the improved group (57 cases, 26 cases) and the stable group (29 cases, 8 cases). SAA increased gradually in the exacerbation group, decreased gradually in the improved group, and slightly increased in the stable group. ΔSAA 3 d were differences among three groups. The change trend of hs-CPR was consistent with SAA. The level of SAA in the death group was higher than that in the survival group on the seventh day. SAA increased in the death group and decreased in survival group with time from hospital admission. There were differences according to ΔSAA 7 d between death group and survival group. The change trend of hs-CPR was consistent with SAA. ROC curve showed that the value of SAA was better than hs-CRP in assessing the severity of patients on admission day, and the area under ROC curve (AUC) was respectively 0.777 [95% confidence interval (95% CI) was 0.669-0.886], 0.729 (95% CI was 0.628-0.830). The value of ΔSAA 3 d was better than SAA on the third day predicting disease trends, and AUC was respectively 0.979 (95% CI was 0.921-1.000), 0.850 (95% CI was 0.660-1.000). hs-CRP on the third day and Δhs-CRP 3 d had no predictive value. Both SAA on the seventh day and ΔSAA 7 d have predictive value for prognosis. AUC was respectively 0.954 (95% CI was 0.898-0.993) and 0.890 (95% CI was 0.689-1.000). SAA on the seventh day and ΔSAA 7 d were better than hs-CRP on the seventh day. Δhs-CRP 7 d have no predictive value. Conclusions:SAA is a sensitive and valuable indicator for CAP patients with viruses and mycoplasma. Dynamic monitoring of SAA can evaluate the patient's progression, prognosis, and assist diagnosis and treatment.
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AIM:To investigate whether Qingshen granules(QSG)-medicated serum inhibits oxidative stress-mediated NF-κB signaling pathway and attenuates epithelial-mesenchymal transition(EMT)of human proximal tubule epi?thelial HK-2 cells induced by high glucose. METHODS:The active components in QSG were analyzed by HPLC. The HK-2 cells were randomly divided into control group,mannitol group,high glucose group,low-dose QSG group,medium-dose QSG group,high-dose QSG group and pyrrolidine dithiocarbamate(PDTC)group. The morphological changes of the cells were observed by inverted phase contrast microscopy. MTT assay was used to detect the cell viability. Flow cytometry was used to detect the content of reactive oxygen species(ROS)in HK-2 cells. ELISA was used to detect the content of malondi?al dehyde(MDA)and the activity of superoxide dismutase(SOD). The DNA binding activity of nuclear factor-κB(NF-κB) p65 in HK-2 cells was detected by electrophoretic mobility-shift assay(EMSA). The protein expression of NF-κB p65, phosphorylated inhibitor of kappa B alpha(p-IκBα),inhibitor of kappa B kinase alpha(IKKα),monocyte chemoattractant protein-1(MCP-1)and intercellular adhesion molecule-1(ICAM-1)in HK-2 cells was detected by Western blot. Immuno?fluorescence staining was used to detect NF-κB p65 andα-smooth mucle actin(α-SMA)protein expression in HK-2 cells. RESULTS:Chlorogenic acid,berberine hydrochloride,plantamajoside,6,7-dimethoxycoumarin,epiberberine,copti?sine,lithospermicacid B,palmatine,leonurine hydrochloride,rheic acid and tanshinone ⅡA in QSG were preliminarily de?termined by HPLC. Compared with control group,the levels of ROS and MDA in HK-2 cells induced by high glucose in?creased(P<0. 05),while the activity of SOD decreased(P<0. 05). The protein levels of NF-κB p65,p-IκBα,IKKα, MCP-1,ICAM-1 andα-SMA were increased(P<0. 05). After intervened by QSG-medicated serum,the levels of ROS and MDA were decreased(P<0. 05),while the activity of SOD was increased(P<0. 05). The protein levels of NF-κB p65,p-IκBα,IKKα,MCP-1,ICAM-1 andα-SMA were decreased(P<0. 05). CONCLUSION:QSG-medicated serum inhibited oxidative stress-mediated NF-κB signaling pathway,thus attenuating the EMT of HK-2 cells induced by high glucose.
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Objective:To explore the application and effect of teaching and anti-teaching system in clinical skills teaching of the standardized residents training.Methods:Forty pediatric residents of Batch 2018 in Hunan Children's Hospital were randomly divided into the conventional teaching group and the teaching and anti-teaching teaching group, 20 in each group. All of them received the skills training for medical licensing examination. A comparative analysis of the teachers' on-site training time, and the time of mastering individual skill, the skill operation ability, and the comprehensive ability in the two groups was conducted. In addition, teachers and students' satisfaction with teaching and anti-teaching system was investigated by Wenjuanxing, a platform providing questionnaire survey. SPSS 17.0 software was used for t-test and Chi-square test. Results:The teachers' on-site training time [(21.75±3.24) min vs. (48.65±2.91) min] and the residents' time of mastering skills and knowledge [(20.46±3.57) min vs. (30.24±4.73) min] in the teaching and anti-teaching teaching group were shorter than those in the conventional teaching group ( P < 0.05). The scores of clinical skills [(92.96±5.21) points vs. (84.13±3.28) points] and the comprehensive ability [(91.23±5.06) points vs. (85.23±5.72) points] of the residents in the teaching and anti-teaching teaching group were higher than those in the conventional teaching group ( P<0.05). The questionnaire survey showed that the residents had a positive attitude towards the teaching and anti-teaching teaching. They believed that it had the advantage of facilitating full-dimensional viewing of the operation, improving the learning interest, and achieving better learning effect for the residents. Conclusion:The application of teaching and anti-teaching system in the standardized training of residents in children's hospitals can make up for the rare adult clinical operation learning opportunities in children's hospitals, rapidly improve the professional skills of residents, improve the passing rate of medical licensing examination, and help to improve the post competency of residents.
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Objective:To observe the effects of "pushing patella and extending knee" manipulation on the protein and mRNA expression levels of integrin β1 (ITGβ1) and phosphorylated adhesion plaque kinase (p-FAK) in rabbit knee osteoarthritis (KOA) model, and to investigate the mechanism of manipulations in the treatment of KOA.Methods:Twenty healthy 6-month-old New Zealand rabbits were divided into the normal group, the model group, the acupuncture group, and the manipulation group according to the random number table method. Among them, the model group, the acupuncture group, and the manipulation group were modeled using the modified Hulth method for KOA. After 7 d of successful modeling, the normal group and the model group did not receive any intervention, while the acupuncture group and the manipulation group received one acupuncture intervention and one "pushing patella and extending knee" manipulation intervention daily, respectively. After 2 weeks of treatment, the rabbit KOA model was executed by air embolization, and the protein and mRNA expression levels of ITGβ1 and p-FAK in knee cartilage were measured by Western blot and real-time polymerase chain reaction (PCR), respectively.Results:Compared with the normal group, the ITGβ1 protein expression level was decreased ( P<0.05) and p-FAK protein expression level was increased ( P<0.05), and the mRNA expression levels of ITGβ1 and p-FAK did not change significantly (all P>0.05) in the model group. Compared with the model group, the ITGβ1 protein expression level was increased ( P<0.05), the p-FAK protein expression level decreased ( P<0.05), and the mRNA expression levels of both ITGβ1 and p-FAK increased (all P<0.05) in the acupuncture group. Compared with the acupuncture group, ITGβ1 protein expression level increased ( P<0.05), p-FAK protein expression level decreased ( P<0.05), and mRNA expression levels of both ITGβ1 and p-FAK increased (all P<0.01) in the manipulation group. Conclusions:The "pushing patella and extending knee" manipulation can optimize the protein and mRNA expression levels of ITGβ1 and p-FAK in the articular cartilage of the rabbit KOA model.
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Objective To investigate the prevalence of Schistosoma japonicum infection in wild mice in Shitai County, Anhui Province, so as to provide insights into precise control of the source of S. japonicum infections. Methods Wild mice were captured using the trapping method for three successive nights at snail-infested settings from Jitan Village of Jitan Township, and Shiquan Village and Xibai Village of Dingxiang Township, Shitai County, Anhui Province in June and October, 2018. All trapped wild mice were sacrificed and liver and mesenteric vein specimens were collected for detection of S. japonicum eggs using microscopy, while the fecal samples in mouse intestines were collected for identification of S. japonicum infections using Kato-Katz technique. In addition, the population density of trapped wild mice was estimated and the prevalence of S. japonicum infection was calculated in trapped wild mice. Results A total of 376 wild mice were trapped from three villages in Shitai County. The population density of trapped wild mice was 9.1% (376/4 124), and the prevalence of S. japonicum infection was 24.2% (91/376) in trapped wild mice. The highest prevalence of S. japonicum infection was detected in Shiquan Village of Dingxiang Township (30.1%), and the lowest prevalence was seen in Xibai Village of Dingxiang Township; however, there was no significant difference in the prevalence of S. japonicum infection in trapped wild mice among three villages (χ2= 4.111, P > 0.05). In addition, there was no significant difference in the prevalence of S. japonicum infection in wild mice captured between on June (26.8%, 34/127) and October (22.9%, 57/249) (χ2 = 0.690, P = 0.406). The trapped wild mice included 6 species, including Rattus norvegicus, Niviventer niviventer, R. losea, Apodemus agrarius, Mus musculus and N. coning, and the two highest prevalence of S. japonicum infection was detected in R. losea (34.9%, 22/63) and R. norvegicus (31.2%, 44/141). Conclusions The prevalence of S. japonicum infections is high in wild mice in Shitai County, and there is a natural focus of schistosomiasis transmission in Shitai County.
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Objective: To analyze the clinical features of patients with acute pulmonary embolism (APE) living in high altitude area of Yunnan province. Methods: This was a cross-sectional retrospective study. APE patients, hospitalized in our hospital between January 2017 and December 2019, were included. The selected patients were divided into low-risk group, medium-risk group and high-risk group according to risk stratification. The clinical data of patients, including demographic data, the main symptoms, risk factors of APE, heart rate and systolic blood pressure and laboratory testing results (D-dimer, cardiac troponin I (cTNI), N terminal B-type natriuretic peptide (NT-proBNP)) and echocardiography and electrocardiogram examination results, were obtained through the electronic medical record system. The clinical characteristics of selected patients were analyzed. Results: A total of 392 patients, aged (63.5±15.7) years, 224 males (57.14%), were included in this study and there were 59 low-risk, 304 medium-risk and 29 high-risk patients in this cohort. The main clinical manifestations were chest pain (157(40.05%)), dyspnea (107(27.30%)), hemoptysis (55(14.03%)), syncope as the first symptom (20(5.10%)), and only 6 cases (1.53%) presented with the typical "Virchow's triad". Most of the patients were accompanied by atypical chest tightness (223(56.89%)) and cough (208(53.06%)). The main risk factors were venous thrombosis of lower limbs (179(45.66%)), hypertension (138(35.20%)), surgery (63(16.07%)), and chronic obstructive pulmonary disease (COPD) (62(15.82%)). There were 57 cases (14.54%) of coronary heart disease, 57 cases (14.54%) of diabetes, 51 cases (13.01%) of cerebral infarction, 47 cases (12.00%) of advanced age, 15 cases (3.83%) of tumor, 7 cases (1.79%) of activity restriction, 6 cases (1.53%) of pregnancy and 4 cases (1.02%) of hormone use in this cohort. The proportion of lower extremity venous thrombosis was significantly higher in low-risk group than in medium-risk group (P<0.01), COPD was more common in high-risk and medium-risk groups than in low-risk group (P<0.01), hypertension was more common in high-risk group than in medium-and low-risk groups (P<0.01). The proportion of advanced age was significantly higher in medium-risk group than in low-risk group (P<0.01). There were no significant differences in RBC and hemoglobin level between low-, medium-and high-risk groups (P>0.05). The level of D-dimer was significantly higher in high-risk group than in medium-and low-risk groups (P<0.05). Levels of NT-proBNP and cTNI were significantly higher in high-risk group than in medium- and low-risk groups (P<0.05). Increased proportion of cTNI and NT-proBNP was significantly higher in high-risk group than in medium- and low-risk groups (P<0.05). There were 105 (26.79%) patients with pulmonary hypertension (PAH). The incidence of PAH was significantly higher in high-risk group than in low-risk group (P<0.01). There were 104 patients (26.53%) with right ventricular enlargement, and the incidence of right ventricular enlargement was significantly higher in high-risk group than in medium-and low-risk groups (P<0.01). Characteristic changes of electrocardiogram in patient with APE were T-wave inversion of limb leads (98(25.00%)), followed by SⅠQⅢTⅢ (83(21.17%)). Conclusions: The main clinical manifestations of APE in Yunnan high altitude area are chest pain and dyspnea, and syncope is the first symptom in some patients, but the typical "Virchow's triad" is rare. The most common risk factors are lower extremity venous thrombosis, hypertension, and COPD. Clinical symptoms, risk factors and laboratory examination results differ among patients with different risk stratification.
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Adolescent , Altitude , Biomarkers , China/epidemiology , Cross-Sectional Studies , Humans , Male , Natriuretic Peptide, Brain , Peptide Fragments , Prognosis , Pulmonary Embolism/epidemiology , Retrospective StudiesABSTRACT
Objective:To explore the application effect of medical waste management system based on B/S architecture in medical waste disposal.Methods:Select a third-class hospital in Yantai for continuous quality improvement of medical waste disposal, and verify the feasibility of the medical waste management system through key indicators such as medical waste related theoretical knowledge, medical waste disposal efficiency, and medical waste disposal management process standardization rate. Through monitoring indicators and result feedback evaluation, it is judged whether the medical waste management system has positive significance for the improvement of medical waste disposal.Results:Taking the adoption of medical waste management system as the node, a five-month comparison before and after was conducted to monitor key indicators such as medical waste disposal efficiency and standardized rate of medical waste disposal management processes. The daily error rate of medical waste classification was reduced from 6.00% to 1.33%, the daily confusion rate of working procedures was reduced from 6.67% to 0.67%, the time delay rate of medical waste disposal was reduced from 7.33% to 2.00%, and the standardized rate of medical waste disposal management processes was increased from 93.33% to 100%. All the data were statistically significant ( P<0.05). Conclusions:Since the operation of the medical waste management system, through the top-down management and control of the administrator and with its unique time retrieval function, the time delay problem of the previous manual management has been greatly improved. The classification management function of medical waste solves the criticism of unclear and chaotic classification of medical waste. The department retrospective function enables the division of responsibilities for medical waste work to be clear, and improves the work efficiency. The construction of medical waste management system based on B/S architecture provided a new information approach for the scientific management of medical waste and a reference for the scientific and effective management of clinical medical waste.
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Objective:To establish a candidate reference procedure for the enumeration of cell particles in urine and applied to the multi-center performance evaluation of an automated urine formed elements analyzer.Methods:According to the standardized mannual microscopic examination of fresh non-centrifuged urine samples and the recommended reference method for enumeration of cell particles in urine published by ISLH, we established a candidate reference procedure for the enumeration of cell particles in urine. From four class A tertiary hospitals′ clinical laboratories, three rigorous trained technicians per hospital tested the same specimen respectively using the reference procedure. Each specimen was repeatedly counted 5 times, obtaining the quantitative results of cell particles were obtained in urine. Four hospitals used the established candidate reference measurement procedure and the automated urine formed elements analyzer to detect 40 to 60 urine specimens from September 2020 to January 2021, and evaluate the established reference method, meanwhile evaluate the accuracy and consistency of the each count from automated urinalysis analyzer.Results:Using the candidate reference measurement procedures, the coefficient of variation of results derived from three trained technicians per hospital was less than 6.98% (red blood cells), 6.99% (white blood cells), 13.94% (epithelial cells) and met the quality requirements. The performance evaluation results of automated urine formed elements analyzer showed that the accuracy of red blood cells, white blood cells and epithelial cells met the requirements (bias≤4.98%) and was well consistent with the reference measurement procedure ( R2≥0.989). Conclusions:A candidate reference measurement procedure for the enumeration of urine cell particles was successfully established with satisfactory precision and accuracy. This procedure was applied to multicenter performance evaluation of an automated urine formed elements analyzer with good accuracy and consistency.
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The clinical data of 4 patients with acute eosinophilic pneumonia (AEP) diagnosed in the Department of Respiratory and Critical Care Medicine, Peking University Third Hospital were retrospectively analyzed. All 4 patients were males, aged 28, 34, 43 and 68 years respectively. The main manifestations were fever, cough and dyspnea. Three cases had a history of smoking and 1 case had a history of ulcerative colitis. The proportion of eosinophils in bronchoalveolar lavage fluid (BALF) was 85%, 93%, 41% and 50%, respectively. The proportion of eosinophils in peripheral blood was 53.3% (16.29×10 9/L), 25.1% (1.29×10 9/L), 4.8% (0.4×10 9/L) and 13.7% (1.55×10 9/L), respectively. Blood gas analysis (without supplemental oxygen) showed type I respiratory failure in 1 case, hypoxemia in 3 cases. The total IgE (normal range ≤ 100.0 KU/L) in peripheral blood of the 4 cases was>2 500.0 KU/L, 60.3 KU/L, 379.4 KU/L and 407.0 KU/L, respectively. HRCT showed diffuse distribution of lesions in both lungs, including ground glass opacity, patchy consolidation and centrilobular nodules. After diagnosis, all patients were treated with systemic glucocorticoids. Fever, cough and dyspnea, as well as the chest CT lesions were significantly improved after treatment. The patients were followed up for 29 months, 16 months, 18 months and 24 months respectively, without recurrence. AEP is a rare disease which is easy to be misdiagnosed as severe pneumonia and/or ARDS. Eosinophilia in peripheral blood is suggestive of the disease, and timely BALF cell differentials is important for early diagnosis.
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Objective:To investigate clinicopathological features of hypopigmented mycosis fungoides (HMF) and hypopigmented interface T-cell dyscrasia (HITCD) .Methods:A total of 41 patients with cutaneous hypopigmented lymphoproliferative diseases, who had complete clinicopathological data, were collected from Department of Dermatology, the Third People′s Hospital of Hangzhou from January 2015 to September 2020, and the clinicopathological and immunophenotypic features were analyzed. Comparisons of normally distributed measurement data were carried out using t test, comparisons of categorical data using Chi-square test or Fisher′s exact test, and comparisons of ranked data between 2 groups using rank-sum test. Results:All of the 41 patients clinically presented with irregular hypopigmentation, some of which was accompanied by erythema or furfuraceous scales. In terms of pathological features, 21 patients showed infiltration and aggregation of atypical lymphoid cells in the epidermis, which was consistent with typical pathological features of mycosis fungoides, and they were diagnosed with HMF; 20 patients showed vacuolar degeneration of the basal layer, accompanied by infiltration of lymphoid cells and mild epidermotropism, and they were diagnosed with HITCD. All immune cells expressed T-cell phenotype, and epidermal lymphocytes expressed a CD8-dominated phenotype in 14 (67%) cases of HMF and 13 (65%) of HITCD. In the epidermis, the total number of lymphocytes was significantly higher in the HMF group than in the HITCD group ( t= 1.81, P= 0.012) ; in the dermis, the number of CD4 + lymphocytes and CD8 + lymphocytes, and the total number of lymphocytes were all significantly higher in the HMF group than in the HITCD group ( t= 2.64, 1.51, 2.60, P= 0.012, 0.002, 0.001, respectively) . All patients were treated with narrow-band ultraviolet B radiation. Among 34 patients who completed the follow-up, 30 achieved complete clearance of skin lesions without recurrence, including all patients with HITCD, and 4 with HMF achieved partial regression of the lesions. Conclusions:Compared with HMF, HITCD presents different pathological characteristics and benign biological behaviors. Thus, HITCD should be distinguished from HMF as an independent disease. Phototherapy alone is effective for the treatment of HITCD.
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ObjectiveTo predict the possible quality markers (Q-markers) of Arisaema Cum Bile in the prevention and treatment of stroke based on ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spetrometry (UPLC-QTOF-MS/MS) and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine (TCMIP) v2.0. MethodUPLC-QTOF-MS/MS was employed with the mobile phase of 0.1% formic acid aqueous solution (A)-0.1% formic acid acetonitrile solution (B) for gradient elution (0-3 min, 0.2%-5%B; 3-5 min, 5%-8%B; 5-8 min, 8%-10%B; 8-14 min, 10%-25%B; 14-18 min, 25%-50%B; 18-20 min, 50%-70%B; 20-21 min, 70%-98%B; 21-23 min, 98%B; 23-24 min, 98%-0.2%B; 24-26 min, 0.2%B), the flow rate of 0.5 mL·min-1 and electrospray ionization (ESI). High quality MS/MS data were scanned in positive and negative ion modes with scanning range of m/z 50-1 500. A local database of the chemical constituents in Arisaema Cum Bile was established by UNIFI 1.8. Then the chemical constituents in Arisaema Cum Bile were characterized by matching with the local database and comparing with the reference substances and literature information. TCMIP v2.0 was used to obtain the targets corresponding to the identified components of Arisaema Cum Bile and stroke, and the "disease-formula" correlation analysis was carried out to screen the core targets by topological eigenvalues. DAVID 6.8 was used for enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway of core targets. According to the "five principles" of Q-markers and combined with literature reports, the Q-markers of Arisaema Cum Bile in the prevention and treatment of stroke were predicted, and the core components acting on these target genes were obtained. Cytoscape 3.8.0 was employed to draw the network diagram of "medicinal materials-active ingredients-target genes-pathways". Finally, AutoDock Vina 1.2.2 was used to calculate and verify the molecular docking between the candidate components and the key targets. ResultA total of 76 chemical components was identified in positive and negative ion modes, 85 core targets were collected for Arisaema Cum Bile in the prevention and treatment of stroke. A total of 31 stroke-related pathways, 23 target genes and 9 main active components of Arisaema Cum Bile acting on these genes were screened, and then we determined 4 possible Q-markers for Arisaema Cum Bile in the prevention and treatment of stroke according to the "five principles". ConclusionThe possible Q-markers of Arisaema Cum Bile for stroke are gallic acid, apigenin-6,8-di-C-glucoside, apigenin and cholic acid, and the target of these four components may be estrogen receptor alpha (ESR1).
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ObjectiveTo observe the effects of Scutellariae Radix (SR)-Paeoniae Radix Rubra (PRR) combination of different proportions on the expression of the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear transcription factor κB (NF-κB) and phosphatidylinositol kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathways in liver tissues of rats with hepatic fibrosis and explore the mechanism against hepatic fibrosis. MethodSixty male SD rats of SPF grade were randomly divided into a normal group, a model group, a positive control (silymarin) group, and SR-PRR 1∶1, SR-PRR 1∶2, and SR-PRR 1∶4 groups, with 10 rats in each group. The hepatic fibrosis model was induced in rats except for those in the normal group by intraperitoneal injection of 40% tetrachloromethane (CCl4)-olive oil solution at 3 mL·kg-1, 5 mL·kg-1 for the first time, for 8 weeks, twice per week. After 4 weeks, rats were treated correspondingly at 10 mL·kg-1 by intragastric administration, and the body weight of rats in each group was weighed for 8 weeks. After administration, histopathological changes in the liver were observed. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), laminin (LN), albumin (ALB), alkaline phosphatase (AKP), and superoxide dismutase (SOD) activities, malondialdehyde (MDA), and hydroxyproline (HYP) content in liver tissues were detected. The mRNA expression levels of TLR4, MyD88, NF-κB, PI3K, Akt, and mTOR in the liver of rats were detected by real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR). ResultCompared with the model group, SR-PRR combination of different proportions could recover the body weight and improve the pathological injury of the liver. As revealed by enzyme linked immunosorbent assay (ELISA) results, compared with the normal group, the model group showed increased ALT, AST, HA, LN, AKP, MDA, and HYP levels to different degrees (P<0.05). Compared with the model group, the groups with drug intervention showed decreased levels of ALT, AST, HA, LN, AKP, MDA, and HYP, potentiated SOD activity, and increased level of ALB (P<0.05). As revealed by Real-time PCR results, compared with the normal group, the model group showed increased mRNA expression of TLR4, MyD88, NF-κB, PI3K, Akt, and mTOR (P<0.05). Compared with the model group, the groups with drug intervention showed reduced mRNA expression of TLR4, MyD88, NF-κB, PI3K, Akt, and mTOR in the liver of rats (P<0.05). ConclusionSR-PRR combination of different proportions can improve the histopathological injury in liver tissues caused by CCl4, with the optimal effect observed in the SR-PRR 1∶4 group. SR-PRR may inhibit the development of liver fibrosis by inhibiting the expression of TLR4/MyD88/NF-κB and PI3K/Akt/mTOR signaling pathways, thereby alleviating chemical-induced liver injury.
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ObjectiveTo explore the molecular mechanism of "transmission between the lung and brain" of influenza based on Janus kinase 1/signal transducer and activator of transcription 1(JAK1/STAT1) signaling pathway and further investigate the intervention effect of Maxing Shigantang (MXSGT). MethodA total of 100 SPF BALB/c mice were randomly divided into a normal group,a model group,an oseltamivir group (21.63 mg·kg-1·d-1),an antiviral granules group(3.9 g·kg-1·d-1), and an MXSGT group(6.05 g·kg-1·d-1), with 20 mice in each group. The pneumonia model was induced in mice except for those in the normal group by intranasal infection of influenza A virus(IAV). Twenty-four hours after modeling,mice were treated with corresponding drugs, while those in the normal group and the model group received the same amount of normal saline by gavage, once a day for 3 and 7 days. The pathological changes in the lung and brain were observed by hematoxylin-eosin(HE)staining. The mRNA expression of IAV nucleoprotein(NP),JAK1, and STAT1 in the lung and brain was detected by real-time quantitative polymerase chain reaction(Real-time PCR), and the protein expression of JAK1 and STAT1 in the lung and brain was detected by Western blot. Immunohistochemical method was used to detect the expression of phosphorylated(p)-STAT1 in the lung and brain tissues, and enzyme-linked immunosorbent assay(ELISA) was used to detect the serum levels of interleukin-1β(IL-1β) and interleukin-10(IL-10). ResultCompared with the normal group, the model group showed obvious pathological changes in the lung tissues and cerebral cortex, increased relative mRNA expression of IAV NP in the lung (P<0.01), elevated mRNA and protein expression of JAK1 and STAT1 in the lung and brain tissues (P<0.05,P<0.01),up-regulated expression level of p-STAT1 in lung tissues and cerebral cortex (P<0.05,P<0.01), and increased serum level of IL-1β (P<0.05). Compared with the model group, the MXSGT group showed alleviated pathological damage to lung tissues and cerebral cortex, decreased relative mRNA expression of IAV NP in lung tissues(P<0.01),reduced mRNA and protein expression levels of JAK1 and STAT1 in lung tissues and brain tissues(P<0.05,P<0.01), and increased serum level of IL-10(P<0.01). ConclusionThe abnormal activation of the JAK1-STAT1 signaling pathway may be one of the molecular mechanisms of "transmission between the lung and brain" of influenza. As an effective compound prescription against the influenza virus,MXSGT can alleviate the pathological damage of brain tissues in mice infected with IAV by regulating the level of cytokines mediated by this pathway.
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ObjectiveTo explore the mechanism of Tongbi Jiangu prescription (TBJG) in the treatment of knee osteoarthritis (KOA) based on network pharmacology and molecular docking,and further verify it by cell experiments. MethodThe active components and the corresponding targets of TBJG were screened out according to the traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP). The targets of KOA were obtained from GeneCards,online mendelian inheritance in man(OMIM), and DrugBank. The common targets of active components of TBJG and KOA were the targets of TBJG against KOA. The active component-target network and protein-protein interaction (PPI) network were constructed by Cytoscape 3.7.2. STRING was used for PPI network analysis. DAVID was used for gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses. Key targets and core active components were selected for molecular docking by AutoDock. The results of network pharmacology were verified by cell experiments and the pharmacodynamic responses were observed. ResultThe prediction of network pharmacology showed that there were 111 active components of TBJG in the treatment of KOA. The core active components were quercetin,kaempferol, and β-sitosterol,and the key targets were interleukin-1β(IL-1β), matrix metalloproteinase-3 (MMP-3),and tumor necrosis factor-α (TNF-α). Biological processes (BP) in GO analysis mainly involved inflammatory response,response to lipopolysaccharide,apoptosis signaling pathway,and regulation of DNA activity in binding transcription factor. Cellular components (CC) included plasma membrane protein complex,RNA polymerase Ⅱ transcription factor complex,membrane raft,and serine/threonine protein kinase complex. Molecular functions (MF) were mainly enriched in cytokine receptor binding,nuclear receptor activity,protein domain specific binding,serine hydrolase activity,chemokine receptor binding,and activity of nitric oxide synthase regulator. As revealed by the KEGG analysis, the relevant signaling pathways were nuclear factor(NF)-κB, Janus kinase(JAK)/signal transducer and activator of transcription(STAT), and Wnt signaling pathways. Molecular docking results showed that the core active components had good binding activities with key targets. The experimental results showed that TBJG could down-regulate IL-1β, MMP-3,TNF-α, and NF-κB p65 expression levels (P<0.05),and up-regulated NF-κB inhibitor(IκB)-α(P<0.05). ConclusionThe mechanism of TBJG in the treatment of KOA lies in the application of active components such as quercetin,kaempferol, and β-sitosterol with IL-1β,MMP-3, and TNF-α as key targets through the NF-κB,JAK/STAT, and Wnt signaling pathways.
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ObjectiveTo observe and analyze the effect of modified Shenling Baizhusan on gastrointestinal dysfunction and protein-energy wasting (PEW) of continuous ambulatory peritoneal dialysis (CAPD) patients with the syndrome of spleen deficiency, blood stasis, and dampness. MethodA total of 66 CAPD patients with the above syndrome were randomized into the observation group and control group, 33 cases in each group. However, 3 cases in each group dropped out, finally leaving 30 cases in each group. Both groups received CAPD and conventional symptomatic treatment. On this basis, the observation group was given modified Shenling Baizhusan (1 bag/day, once in the morning and again in the evening, 12 weeks), and the control group the bifidobacterium capsules (1.05 g/time, twice/day, 12 weeks). Before and after treatment, the traditional Chinese medicine (TCM) syndrome score, gastrointestinal symptom rating scale (GSRS) score, and malnutrition-inflammation score (MIS) in two groups were recorded, and the levels of serum albumin (ALB), prealbumin (PA), transferrin (TRF), gastrin-17 (G-17), tumor necrosis factor alpha (TNF-α), interferon-γ (IFN-γ), and interleukin-10 (IL-10) were detected. Moreover, body mass index (BMI) was calculated. ResultAfter treatment, the alleviation of the TCM syndrome in the observation group was better than that in the control group (Z=-2.591, P<0.05), and the TCM syndrome score in the observation group was lower than that in the control (P<0.05). The symptom scores, MIS, and G-17 of the observation group were significantly decreased compared with those before observation and in the control group (P<0.05). After treatment, the GSRS scores of the two groups were significantly lower than those before treatment (P<0.05), particularly the observation group (P<0.05). ALB, PA, TRF, and BMI of the observation group after treatment were increased compared with those before treatment and those of the control group after treatment (P<0.05). After treatment, serum TNF-α and IFN-γ of the two groups were significantly reduced compared with those before treatment (P<0.05), and the levels of the two in the observation group were significantly lower in the observation group than in the control group (P<0.05). After treatment, IL-10 level of the observation group was higher than that before treatment and in the control group (P<0.05). ConclusionThe modified Shenling Baizhusan can relieve the gastrointestinal dysfunction and PEW in CAPD patients with the syndrome of spleen deficiency, blood stasis, and dampness.
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ObjectiveTo study the correlation between the apparent color, comprehensive sweetness and the content of main components in the preparation of Rehmanniae Radix Praeparata processed with Amomi Fructus and Citri Reticulatae Pericarpium, so as to lay a foundation for revealing the processing principle of Rehmanniae Radix. MethodThe color of Rehmanniae Radix Praeparata sample powder was measured by automatic colorimeter, the contents of 14 active components in samples with different heating time points were determined by high performance liquid chromatography, including 7 glycosides of catalpol, rehmannia glycoside D, leonurine glycoside, 5-hydroxymethyl furfural, verbascoside, isoacteoside and hesperidin, and 7 carbohydrates of D-fructose, glucose, sucrose, melibiose, raffinose, manninotriose and stachyose), and the mobile phase was acetonitrile-water for gradient elution. The comprehensive sweetness difference of sample was calculated by the sweetness of saccharides, SPSS 21.0 was used to analyze the relationship between the color, comprehensive sweetness and the main component contents in the processing of Rehmanniae Radix Praeparata processed with Amomi Fructus and Citri Reticulatae Pericarpium, the quality comprehensive evaluation index of Rehmanniae Radix Praeparata by triangular area method was established. ResultDuring the processing, the color value of the powder increased, and the apparent color of the sample became darker. the content determination results showed that the content of glycosides decreased, monosaccharides and comprehensive sweetness increased with the increase of heating time. The results of correlation analysis showed that chromaticity value, comprehensive sweetness were significant negatively correlated with the content of iridoid glycosides (P<0.01), the chromaticity value was significant positively correlated with the contents of furaldehyde derivatives, phenylethanoid glycosides, flavonoids and comprehensive sweetness was significant positively correlated with the contents of furaldehyde derivatives, phenylethanoid glycosides (P<0.01), and the comprehensive sweetness was positively correlated with the content of flavonoids (P<0.05). After 52 h of processing, the comprehensive evaluation index of samples reached 0.99. ConclusionThe overall trend of cluster analysis of powder chromaticity value of Rehmanniae Radix Praeparata is basically consistent with that of naked eyes, the comprehensive quality evaluation of Rehmanniae Radix Praeparata processed with Amomi Fructus and Citri Reticulatae Pericarpium can be carried out by combining the three indexes of powder chromaticity value, comprehensive sweetness and glycosides content.
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Pathogenic bacterial infection is one of the main clinical symptoms. Antibiotics are widely used in clinical practice to inhibit or kill the bacteria, fungi and other pathogenic microorganisms. However, with the massive use of antibiotics, drug-resistant strains continue to appear that make the antibacterial situation is becoming increasingly severe. Due to the advantages of multiple targets, multiple pathways and multiple components, traditional Chinese medicine (TCM) have gradually attracted more attention and were used in antibacterial treatment. However, some antimicrobial TCM have problems such as low solubility, poor stability, and low bioavailability. Improving and enhancing the antibacterial activity of TCM through preparation technology is one of the effective solutions. Based on this, two aspects of unilateral antibacterial TCM preparation technology and combination antibacterial preparation technology are introduced, including inclusion technology, nanotechnology, electrospinning, 3D printing and others. Distinctive features and specific application effects of these preparation technologies are explained firstly, and then their advantages and disadvantages are compared and analyzed. The review can be a useful reference for improving the antibacterial activity of TCM.
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Sleep plays an important role in energy balance. As reported, sleep disorder is an important risk factor for metabolic diseases. Controlling the relationship between energy metabolism and sleep can affect sleep homeostasis and body metabolic rate. Chinese medicine, with remarkable curative effects in the prevention and treatment of insomnia, has the characteristics of green, safety, and few side effects, and attracts extensive attention of scholars in the world. In recent years, remarkable progress has been made in the research on the mechanism of Chinese medicine in interfering with sleep. This paper reviewed the research progress of mind-tranquilizing Chinese medicines, such as compounds (pterostilbene), Chinese medicinal drugs (Ziziphi Spinosae Semen), and Chinese medicinal prescriptions (Jiaotaiwan, Suanzaoren tang, Tianwang Buxindan, Anmeidan, Banxia Houpotang, Qihuo decoction, Songyu Anshen prescriptions, and Shuxie Yihao prescriptions) in the treatment of sleep disorders by regulating energy metabolism. The findings revealed that Chinese medicine can intervene in the sleep deprivation model by affecting metabolism-related pathways such as material metabolism, mitochondrial function, oxidative stress and inflammatory response, appetite system, and biological clock system. In terms of frequency of use, the top drugs are Ziziphi Spinosae Semen, Poria, Schisandrae Chinensis Fructus, and Salviae Miltiorrhizae Radix et Rhizoma which affect heart and liver meridians to regulate blood circulation, ensure energy supply, and play the role of nourishing the heart and tranquilizing the mind. The present paper summarized the effects and mechanisms of Chinese medicine in the treatment of insomnia and other sleep disorders from the perspective of energy metabolism to provide references for further research and exploration of diseases in the future.