Objective To study a nucleic acid extraction method suitable for detecting methicillin‐resistant Staphylococcus aureus (MRSA) by PCR method .Methods Under different incubation conditions ,MRSA was cracked by lysozyme ,lysostaphin or chel‐ex100R resin for obtaining DNA ,then the target gene was detected by using the PCR method .Results DNA was obtained by sim‐ultaneously using lysozyme ,lysostaphin and chelex100R resin solution ,the obtained Ct value was significantly lower than that of the other components of schizolysis solutions when PCR was used to detect mecA gene of obtained DNA .There was no statistically sig‐nificant difference between adopting the 56 ℃ one‐step method and the 37 ℃ and 56 ℃ two‐step method for conducting MRSA schizolysis(P> 0 .05) ,but the steps were simplified .Conclusion Incubating MRSA in solution containing lysozyme ,lysostaphin , chelex100R resin for 30 min at 56 ℃ is the convenient and efficient schizolysis method to extract DNA ,which can be used immedi‐ately for the next step of PCR and lays the foundation for PCR rapid detection of clinical MRSA infection .

ObjectiveThis clinical study is aimed to investigate whether levetiracetam (LEV) can improve electrocorticogram (EEG) in epileptic children epilepsy patients with better clinical manifestation but abnormal EEG findings.MethodsTotally 39 children from our neurological clinic with partial or complex partial epilepsy seizure were included in present study and assigned equally into three groups receiving different treatment:control group,sodium valproate (VPA) group,and LEV group.Their clinical symptoms had been controlled for over one year by carbamazepine ( CBZ),but EEG results showed clearly abnormal.Epileptiform discharges were observed in routine EEG exams half a month before recruiting.After recruiting,they continued to receive CBZ alone (control group) or co-treated with VPA ( VPA group) or LEV (LEVgroup),respectively.Six months later,EEG was taken again and results were analyzed.ResultsImprovement rate were 9.1％ ( control group),23.1％ ( VPA group),and 66.7％ (LEV group),respectively;Overall statistical difference was reached among three groups ( P＜0.01 ) and between control group and LEV group( P＜0.012 5 ),but no statistical difference between control group and VPA group was reached ( P＞0.0125).ConclusionCo-treatment of LEV in child epilepsy patients receiving CBZ can significantly decrease abnormal EEG discharge frequency during interictal period.

Objective: To observe the effects of Danggui Shaoyao powder (DSP) on hepatic lipid metabolism and further explore its mechanism of action by peroxisome proliferator-activated receptor (PPARγ)-liver X receptor (LXRα)-adenosine triphosphate (ATP)-binding cassette transporter A1 (ABCA1) pathway regulation. Methods: Eight C57BL/6J male mice were selected as the control group, and 24 ApoE−/− male mice were randomly divided into the atherosclerosis model (AS) group, atorvastatin calcium (AC) group, and DSP group (n = 8 each group). To establish an AS model, ApoE−/− mice were fed a high-fat diet for 16 weeks. Pathologic changes in the aortic vasculature and liver were identified using Oil Red O staining. Triglyceride (TG), cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C) levels were determined in the livers using a single-reagent GPO-PAP method. Fluorescence quantitative polymerase chain reaction and western blot were used to observe and evaluate the mRNA and protein expression of the PPARγ-LXRα-ABCA1 intermediates in the liver. Results: After 16 weeks of a high-fat diet, ApoE−/− mice showed more Oil Red O staining in the aorta and liver compared to the CONT group. Compared to the AS group, the DSP and AC treatment reduced aortic plaque and hepatic lipid deposition to varying degrees. Furthermore, DSP significantly reduced the hepatic lipid area in ApoE−/− mice (P < .001) and decreased the levels of TG, TC, and LDL-C in liver (P < .001, P = .027, P < .001, respectively). DSP also significantly increased the levels of PPARγ, LXRα, ABCA1, and ABCG1 mRNA expression, as well as the PPARγ, LXRα, ABCA1, and ABCG1 protein expression in liver. Conclusion DSP improved hepatic lipid metabolism via PPARγ-LXRα-ABCA1 pathway modulation for AS treatment.

Objective To measure the expression of circulating microRNA (miRNA)in patients with hepatitis B virus (HBV)-related liver failure and its relationship with disease prognosis.Methods The miRNA expressions in serum of 5 patients with HBV-related liver failure and 5 healthy control subjects were compared using Exiqon miRCURY LNATM miRNA microarray.The sera from 20 patients with chronic hepatitis B (CHB),20 patients hepatitis B related cirrhosis,50 patients with HBV-related liver failure and 40 healthy persons in Ruijin Hospital were collected.The relative expression of miRNA-595 was measured using quantitative real-time polymerase chain reaction (PCR).The relative expressions of miRNAs among groups were analyzed using student t test,the correlations were analyzed by Pearson and Spearman correlation.Results Microarray informed that 92 miRNAs changed significantly in patients with HBV-related liver failure,and miRNA-595 increased most significantly.The results of real-time PCR showed that the relative expressions of miRNA-595 ,miRNA-300 and miRNA-122 were 6.03 (t=3.134, P =0.003),3.12 (t=7.221 ,P <0.01)and 2.77 (t=2.671 ,P =0.021),which were higher compared to those in healthy control group.In the analysis of the relationship between miRNA-595 expression and disease prognosis in patients with HBV-related liver failure,the relative expressions of miRNA-595 in patients with CHB,hepatitis B related cirrhosis and HBV-related liver failure were 2.26 (t =3.780,P =0.001),3.32 (t = 6.111 ,P < 0.01)and 6.03 (t = 3.134,P = 0.003),respectively,which were all increased compared to that of the healthy control.The relative expression of miRNA-595 of patients with HBV-related liver failure was 2.66 times (t=2.450,P =0.043)higher than that of patients with CHB. When dividing patients according to prothrombin activity,miRNA-595 increased significantly in patients with early stage liver failure.When dividing patients according to model of end-stage liver disease (MELD) score,MELD score was positive correlated with the expression of miRNA-595 when MELD score was under 30 (r=0.673,P =0.004).The expression of serum miRNA-595 in survival group (11 .08,n=23) was higher than that in non-survival group (3.67,n = 27,t =4.309,P =0.041).Conclusions The expressions of miRNA595 ,miRNA-300 and miRNA-122 are all increased in patients with HBV-related liver failure,especially the expression of circulating miRNA-595 at early stage of the disease.The miRNA-595 may be used as a new serum biomarker for monitoring the severity of disease.

Objective:To carry out the Meta-integration of Intensive Care Unit (ICU) nurses' real experience of palliative care.Methods:The qualitative study that met the criteria in the Cochrane Library, PubMed, EMbase, Web of Science, China Biology Medicine Literature Service System, China National Knowledge Infrastructure (CNKI) , VIP and Wanfang Data were searched by computer from the establishment of the database to June 30, 2020. The quality of the included literature was evaluated using the critical appraisal checklist for qualitative researches of the Australia Joanna Briggs Institute (JBI) Evidence-based Health Center (2017 edition) . The collective integration method was used to integrate the results.Results:A total of 17 articles were included, involving 16 in English and 1 in Chinese. Finally, it was summarized into 11 categories, and 4 integrated results were collected, which were the complex palliative care (multiple roles of ICU nurses in palliative care, patient care, and family care) ; insufficient preparation for palliative care (lack of professional skills, communication skills and psychological preparation) ; insufficient support from palliative care organizations (inadequate psychological support, multidisciplinary team cooperation, and lack of professional palliative care service systems) ; insufficient legal support for palliative care (lack of decision-making power for palliative care, ethics and legal issues in palliative care) .Conclusions:ICU nurses lack the ability to palliative care and lack a reliable support system. Managers should pay attention to the negative experience and needs of ICU nurses for palliative care, and gradually carry out education and training to improve the level of care and the quality of life of dying patients.

Objective To evaluate the efficacy and safety of Qingpeng ointment for the treatment of localized eczema in children.Methods A multicenter,randomized,open-labeled,tacrolimus 0.03％ ointment-controlled clinical trial was conducted.A total of 442 children with localized eczema were enrolled into this study,and divided into 2 groups to topically apply Qingpeng ointment (Qingpeng group) and tacrolimus 0.03％ ointment (tacrolimus group) respectively twice a day for 2 weeks.The children were followed up before the treatment and 1,2 weeks after the treatment,and the efficacy and safety were evaluated at the same time.Results Clinical data from 426 children were included in the efficacy analysis.At 1,2 weeks after the treatment,there were no significant differences in the response rate between the Qingpeng group and tacrolimus group (1 week after the treatment:78.92％ vs.81.77％,x2 =0.545,P =0.460;2 weeks after the treatment:98.65％ vs.97.54％,x2 =0.721,P =0.396).However,the pruritus scores at 1,2 weeks after the treatment were both significantly lower in the Qingpeng group than in the tacrolimus group (1 week:F =14.001,P =0.000;2 weeks:F =11.252,P =0.001).At 1 week after the treatment,the incidence rate of adverse reactions was significantly lower in the Qingpeng group than in the tacrolimus group (P ＜ 0.05).Mild itching,burning sensation and other local irritant sensations were the most common adverse reactions.Conclusion Qingpeng ointment is safe and effective for the treatment of localized eczema in children with good tolerability.

Objective:To investigate the clinicopathological features, treatment programs and prognosis of patients with primary diffuse large B-cell lymphoma (DLBCL) in cavernous sinus.Methods:The clinical data of a patient with primary DLBCL in cavernous sinus who were admitted to Wuhan No.1 Hospital in December 2020 were retrospectively analyzed, and the relevant literature was reviewed.Results:The patient was a 63-year-old female who underwent resection of the cavernous sinus lesion, and the pathological diagnosis was DLBCL. The patient received 6 courses of R-CHOP regimen chemotherapy, lumbar puncture + intrathecal injection of chemotherapy drugs, and twice additional rituximab immunochemotherapy, and no tumor cells were found in the results of liquid-based thin layer cytology for cerebrospinal fluid exfoliated cells; twice magnetic resonance imaging (MRI) re-examination after the operation showed no recurrence and adjacent metastasis of the tumor. The patient's symptoms were significantly improved without residual neurological sequelae.Conclusions:Primary DLBCL in cavernous sinus is rare in clinical practice, early diagnosis is crucial for the prognosis of patients, and different protein expression may indicate the prognosis. Biopsy, complete resection of the tumor under the premise of preserving important anatomical structures and functions, and standardized chemotherapy combined with intrathecal injection local chemotherapy can effectively prolong the survival time of patients and improve the quality of life.

Objective:The cellular immunity of 5 Mycobacterium tuberculosis recombinant proteins and their compositions was evaluated.Method:A total of 88 fresh venous blood from peripheral heparin anticoagulant population, 42 of which were from tuberculosis patients treated by The Tuberculosis Prevention and Treatment Center of Changping District, Beijing, and 46 of healthy volunteers were provided by the Infection Diseases of Chinese Center for Disease Control and Prevention. Healthy volunteers without a history of tuberculosis exposure and any clinical signs and symptoms. Using the Mycobacterium tuberculosis standard strain H37Rv DNA as a template, complete genes of the selected 5 recombinant proteins Rv3874, Rv3875, Rv2031c, Rv1411c and Rv3418c by PCR amplified; 5 recombinant proteins were cloned, expressed and purified as stimulants by genetic recombination and protein purification techniques, and the effector T cell enzyme-linked immunospot assay (ELISPOT) was used to detect cellular immunity in the population.Results:The recombinant proteins Rv3874, Rv3875, Rv2031c, Rv1411c and Rv3418c were successfully cloned, expressed and purified; And the sensitivities were 50.00%, 71.43%, 69.04%, 73.81% and 76.19%, and the specificities were 86.96%, 76.09%, 71.74%, 39.13% and 36.96%. In addition, the positive predictive value, negative predictive value, area under the curve and Youden index were 52.46% to 77.78%, 62.96% to 74.47%, 0.511 to 0.754 and 0.129 to 0.475, respectively. Except for Rv1411c and Rv3418c, the number of spot-forming cell (SFC) detected by Rv3874, Rv3875 and Rv2031c in tuberculosis patients was higher than healthy volunteers, and the differences were statistically significant ( P<0.001). Among the 26 compositions composed of 5 recombinant proteins, the sensitivity was 80.95% to 95.24%, and the specificity was 68.89% to 24.44%. As the number of recombinant proteins in the composition increases, the sensitivity gradually increased, but the specificity decreased. Conclusion:The recombinant proteins of Mycobacterium tuberculosis Rv3874, Rv3875 and Rv2031c have strong ability to stimulate T cells to produce immune response, and have certain antigenicity. The efficacy of Rv1411c and Rv3418c alone as diagnostic antigens is not ideal, and the composition composed of multi-component antigens has certain application value. This article provides experimental evidence for the immune diagnosis of tuberculosis and the preparation of new anti-tuberculosis vaccines.

Objective To describe and analyze the current situation of the four same type of departments in an hospital in order to provide a reference for the construction of"the most cost-effective medical care".Methods The CN-DRG were used to automatically group and compare the medical capacity and inpatient service efficiency of the hospital department groups,and in the refined analysis,one DRG disease group of in situ cancer and non-malignant disease loss uterine surgery and single species uterine fibroid was included,and the Kruskal-Wallis H test was used to further compare the differences in length of stay and various costs.Results It included a total of 22630 patients,whose weights varied from a maximum of 3948.62 in diagnostic group 1 to a minimum of 133.55 in diagnostic group 11.The cost consumption indexes ranged from a minimum of 0.89 in diagnostic group 5 to a maximum of 1.04 in diagnostic group 2,while the time consumption indexes ranged from a minimum of 0.48 in diagnostic group 11 to a maximum of 0.81 in diagnostic group 5.When comparing the diagnostic groups,there were statistically significant differences(P＜0.05)in hospitalization days,total cost,diagnostic cost,therapeutic cost,and cost of supplies.Specifically,when comparing the diagnostic and treatment groups within departments,the differences in hospitalization days and all costs were statistically significant(P＜0.05)in departments 1 and 2,the differences in diagnostic cost,therapeutic cost,and cost of supplies were statistically significant(P＜0.05)in department 3.Conclusion There exists a notable disparity in the extent to which each diagnostic and treatment group contributes to the hospital's service capacity and cost variability.Consequently,it is necessary to reasonably evaluate the length of hospital stay and medical cost of patients to achieve the highest cost-effective medical treatment.

Objective:The cellular immunity of 5 Mycobacterium tuberculosis recombinant proteins and their compositions was evaluated.Method:A total of 88 fresh venous blood from peripheral heparin anticoagulant population, 42 of which were from tuberculosis patients treated by The Tuberculosis Prevention and Treatment Center of Changping District, Beijing, and 46 of healthy volunteers were provided by the Infection Diseases of Chinese Center for Disease Control and Prevention. Healthy volunteers without a history of tuberculosis exposure and any clinical signs and symptoms. Using the Mycobacterium tuberculosis standard strain H37Rv DNA as a template, complete genes of the selected 5 recombinant proteins Rv3874, Rv3875, Rv2031c, Rv1411c and Rv3418c by PCR amplified; 5 recombinant proteins were cloned, expressed and purified as stimulants by genetic recombination and protein purification techniques, and the effector T cell enzyme-linked immunospot assay (ELISPOT) was used to detect cellular immunity in the population.Results:The recombinant proteins Rv3874, Rv3875, Rv2031c, Rv1411c and Rv3418c were successfully cloned, expressed and purified; And the sensitivities were 50.00%, 71.43%, 69.04%, 73.81% and 76.19%, and the specificities were 86.96%, 76.09%, 71.74%, 39.13% and 36.96%. In addition, the positive predictive value, negative predictive value, area under the curve and Youden index were 52.46% to 77.78%, 62.96% to 74.47%, 0.511 to 0.754 and 0.129 to 0.475, respectively. Except for Rv1411c and Rv3418c, the number of spot-forming cell (SFC) detected by Rv3874, Rv3875 and Rv2031c in tuberculosis patients was higher than healthy volunteers, and the differences were statistically significant ( P<0.001). Among the 26 compositions composed of 5 recombinant proteins, the sensitivity was 80.95% to 95.24%, and the specificity was 68.89% to 24.44%. As the number of recombinant proteins in the composition increases, the sensitivity gradually increased, but the specificity decreased. Conclusion:The recombinant proteins of Mycobacterium tuberculosis Rv3874, Rv3875 and Rv2031c have strong ability to stimulate T cells to produce immune response, and have certain antigenicity. The efficacy of Rv1411c and Rv3418c alone as diagnostic antigens is not ideal, and the composition composed of multi-component antigens has certain application value. This article provides experimental evidence for the immune diagnosis of tuberculosis and the preparation of new anti-tuberculosis vaccines.