Objective To introduce comet assay and to analyze the DNA damage and repair ability of human fetal lung diploid fibroblasts (2BS) as well as lymphocytes. Methods The mixture of 2BS cells (or lymphocytes) and 1% LMA is put on the gel of 0. 5% NMA. After breakage, electrophoresis and stain, the comet tail distance and comet tail area of 100 cells (per sample) are analyzed with Leica image analysis system. Results There are no obvious comet tail for most of young and senescent 2BS cells as well as lymphocytes from young and senescent donors. The comet tails of 28PD 2BS cells treated with H2O2 are obvious. Moreover, the comet tail distances and tail areas of the cells increase as the concentration of H2O2 increasing. The stastics data of the 28PD 2BS cells exposed to H2O2 are as following. Comet tail distance: s = 1.7 ~3. 5 (?m)、CV = 1. 6% ~ 2. 8% . Comet tail area: s = 1. 5% ~ 2. 3%、CV = 1. 8% ~ 2. 5% . Conclusions The DNA breakage level is low both for young and senescent cell. H2O2 can damage DNA obviously. Comet assay is one of important methods used to comment on DNA damage and DNA repair ability.

ObjectiveTo investigate the impacts of sepsis-induced cardiac dysfunction on hemodynamics, organ function and prognosis in the patients with septic shock.Methods A prospective cohort study was conducted in 44 patients suffering from septic shock with the duration< 24 hours admitted to the Department of Critical Care Medicine of Peking University Third Hospital during June 2013 to June 2014. The patients were divided into two groups according to the left ventricular ejection fraction (LVEF) as recorded in echocardiogram at time of admission to the intensive care unit (ICU) as sepsis-induced myocardial dysfunction group (LVEF< 0.50,n= 11) and normal cardiac function group (LVEF≥0.50,n= 33). The cardiac function evaluation and hemodynamics monitoring were performed with echocardiogram and pulse-induced contour cardiac output (PiCCO) on 1, 3, 7 days after the ICU admission. The plasma levels of the biomarkers of myocardial damage, troponin T (TnT) and N-terminal pro-brain natriuretic peptide (NT-proBNP) were measured, and the parameters representing organ function and the 28-day prognosis were collected as well.Results On the ICU admission, central venous pressure (CVP) and left ventricular end-diastolic diameter (LVEDD) were obviously lower in normal cardiac function group than those of myocardial dysfunction group [CVP (mmHg, 1 mmHg = 0.133 kPa): 10±4 vs. 14±6,P< 0.05; LVEDD (mm): 45.0±5.3 vs. 51.8±7.1,P< 0.01], and there was no significant difference in other hemodynamic parameters between two groups. On the 3rd day, all the cardiac function and hemodynamic parameters showed no significant differences between the two groups. On the 7th day, the cardiac index (CI) and pulmonary vascular permeability index (PVPI) of normal cardiac function group were significantly higher than those of myocardial dysfunction group [CI (mL·s-1·m-2): 63.3±13.3 vs. 48.3±10.0,P< 0.05;PVPI: 1.5 (1.4, 1.9) vs. 1.1 (0.7, 1.1),P< 0.01], and no significant difference was found in the other parameters. The plasma levels of TnT and NT-proBNP were found to have no difference at three time points between two groups. There was no difference in the number or the extent of organ dysfunction, including lung, kidney, liver and coagulation system, between the groups at the time of ICU admission. There was no obvious difference in the 28-day survival rate between the myocardial dysfunction group and normal cardiac function group [81.8% (9/11) vs. 72.7% (24/33),χ2= 0.398, P= 0.528].Conclusions Sepsis-induced myocardial dysfunction is a reversible organ dysfunction. It can directly induce decreased left ventricular systolic function and enlargement of ventricle in patients with septic shock without reducing cardiac output or impairing the functions of other organs, or elevating the mortality rate.

OBJECTIVE To study the isolation status and antimicrobial resistance of nonfermenting Gram-negative bacilli collected from intensive care unit(ICU) of our hospital so as to instruct the rational clinical application of antibiotics.METHODS The antimicrobial resistance of nonfermenting Gram-negative bacilli isolates collected from patients in ICU from Jan 2003 to Dec 2007 was analyzed.Antimicrobial susceptibility of clinical isolates were tested by Kirby-Bauer method.RESULTS Total 384 nonfermenting Gram-negative bacilli isolates were collected in 5 years.The most common species were Acinetobacter baumannii(219),Pseudomonas aeruginosa(117) and Stenotrophomonas maltophilla(36).The antimicrobial resistance rate of nonfermenting Gram-negative bacterial to most antibiotics were much higher.The antimicrobial resistance rate of Acinetobacter spp to imipenem,cefoperazone/sulbactam and piperacillin/tazobactam was 3.7%,28.3% and 42.9%.But the resistance rate of Acinetobacter spp to imipenem was increased in recent 2 years(58.0%).The antimicrobial resistance rate of P.aeruginosa to cefoperazone/sulbactam was the lowest.That of imipenem-resistant P.aeruginosa to cefoperazone/sulbactam was 34.0%.S.maltophilla was relatively susceptible to ceftazidime,cefoperazone/sulbactam and piperacillin/tazobactam.CONCLUSIONS Nonfermenters Gram-negative bacilli are the important pathogens in ICU.Surveillance of their prevalence and drug resistance may provide evidences for rational antibiotic choices.

Objective To observe the treatment response and potential issues related to antiretroviral therapy (ART) in pediatric patients with acquired immunodeficiency syndrome (AIDS) and to provide a basis for revising the treatment guideline and improving the clinical practice. Methods Children eligible for ART according to the current treatment guideline were recruited from Dehong area. Enrolled patients were provided with ART and followed up for regular clinical check and laboratory tests. Results By the end of March 2009, a total of 70 children had received ART. Among them, 60 patients were treated with regimen including zidovudine (AZT)+ lamivudine (3TC)+nevirapine (NVP). Twelve, eighteen, twenty-three and nineteen patients have tested for HIV viral load after 3 month, 6 month, 12 month and 18 month treatment, respectively. Among them, 7, 12,14 and 14 patients respectively achieved HIV viral load lower than 1000 copy/mL. Average CD4+ Tlymphocyte count increased from (317.8±288.8) × 106/L at baseline to (1037.2±1086. 1) × 106/Lafter 18 month treatment. Side effects mainly occurred within the first 3 months of treatment. Nearly 50% of children had gastrointestinal symptoms. Resistance to 3TC, NVP and efavirenz (EFV) were found in five patients who have completed 12 months of treatment. Conclusions Pediatric AIDS patients show good compliance and treatment response to ART. Most side effects happen in the first 3months of treatment and the most common side effects are gastrointestinal symptoms.

Objective:To observe the distribution of vascular endothelial growth factor (VEGF) and microvessel density (MVD) in different brain regions in aged rats and determine the role of VEGF and MVD in the aging process of the nervous system. Methods:We observed the expression of VEGF and MVD in different parts of rat brain in the 3- month group and 30-month group with immunohistochemical technique. Results:Compared with the 3-month group, the 30-month group showed fewer VEGF-positive cells and MVD in the brain (P<0.01), and the number varied signiifcantly in different brain regions(P<0.01). The motor cortex region contained more VEGF-positive cells and MVD than the hippocampus and cerebellum. Conclusion:VEGF-positive cells and MVD are decreased in every brain region of aged rats, and the motor cortex region contains more positive cells, suggesting exogenous VEGF may enhance the formation of microvessels and delay the aging of the nervous system.

Objective:To compare low temperature coblation assisted tonsillectomy with conventional dissec-tion tonsillectomy intraoperation and afteroperation. Method:Ninty-two pedia-patients were randomly divided into coblation-assisted tonsillectomy group and the control group. Arthrocare EVac 70 T&.A Wand was used for cobla-tion-assisted tonsillectomy, and the energy set was 7. Conventional dissection tonsillectomy was operated in control group. The operating time,bleeding,tonsillar fossae healing and return to normal diet and activities, intraoperative complications were recorded seperately. Postoperative pain of the pedia-patients was recorded daily using Wong Baker FACES pain scale for 10 days of afteroperation. Result: Coblation assisted group had a shorter operative time than the control group (10. 2min vs. 36. 5min, P<0. 001). The average amount of intraoperative bleeding of Coblation assisted group was(6. 83±3. 36) ml, while the control group was(30. 07±7. 04)ml. Pedia-patients of Coblation group had a better pain score on 1, 2, 3 days after operation than in control group, and there was no statistically significant difference on 4 to 10 days between the two groups. Coblation group ones returned to normal diet earlier than the control group, but both group required similar time returned to normal activities. There was one patient in control group, who required suturing to stop bleeding for active oropharyngeal bleeding on the 6th day after operation. Conclusion: Comparing with traditional dissection, Coblation assisted tonsillectomy is easy to perform with less intraoperative bleeding,shorter operation time,earlier return to normal diet, less pain on 1st to 3th day postoperatively.

ObjectiveTo investigate the anti-liver cancer mechanism of water extract of Jianpi Xiaoji prescription and its effect on cell autophagy flow and autophagy-related proteins. MethodsSMMC7721 cells were treated with water extract of Jianpi Xiaoji prescription. CCK-8 assay was used to measure cell proliferation; flow cytometry was used to measure cell apoptosis rate; Western blot was used to measure the expression of the autophagy-related proteins Beclin1, LC3, and P62; the formation of autophagosomes and the fusion of autophagosomes and lysosomes were monitored. The independent samples t-test was used for comparison of continuous data between two groups; a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsThe CCK-8 assay showed that cell proliferation was significantly inhibited at 24, 48, 72, and 96 hours after the drug was added, and there was a significant difference between the drug treatment group and the control group (t=28.458, 81.093, 85.328, and 100.158, all P＜0.001). Flow cytometry showed that compared with the control group, the drug treatment group had a significant increase in the apoptosis of liver cancer SMMC7721 cells (t=-42.629, P＜0.001). Western Blot showed that compared with the control group, the drug treatment group had significantly downregulated expression of Beclin1 and significantly upregulated expression of the autophagy markers LC3-II and P62. The flow of autophagosomes to autolysosomes was blocked in the drug treatment group, and there was a significant difference between the drug treatment group and the control group (F=31.155, P＜0.001). ConclusionThe water extract of Jianpi Xiaoji prescription can inhibit the proliferation of human liver cancer SMMC-7721 cells and promote apoptosis; meanwhile, it can upregulate the expression of Beclin1, downregulate the expression of LC3 and P62, and block the formation of autophagy flux.

OBJECTIVE: To compare low temperature coblation assisted tonsillectomy with conventional dissection tonsillectomy intra-operation and after-operation. METHOD: Ninety-two pedia-patients were randomly divided into coblation-assisted tonsillectomy group and the control group. Arthrocine EVac 70 T&A Wand was used for coblation-assisted tonsillectomy, and the energy set was 7. Conventional dissection tonsillectomy was operated in control group. The operating time,bleeding,tonsillar fossae healing and return to normal diet and activities, intraoperative complications were recorded separately. Postoperative pain of the pedia-patients was recorded daily using Wong Baker FACES pain scale for 10 days of after-operation. RESULT: Coblation assisted group had a shorter operative time than the control group (10.2 min vs. 36.5 min, P<0.001). The average amount of intraoperative bleeding of Coblation assisted group was (6.83+/-3.36) ml, while the control group was (30.07+/-7.04) ml. Pedia-patients of Coblation group had a better pain score on 1, 2, 3 days after operation than in control group, and there was no statistically significant difference on 4 to 10 days between the two groups. Coblation group ones returned to normal diet earlier than the control group, but both group required similar time returned to normal activities. There was one patient in control group,who required suturing to stop bleeding for active oropharyngeal bleeding on the 6th day after operation. CONCLUSION Comparing with traditional dissection, Coblation assisted tonsillectomy is easy to perform with less intraoperative bleeding, shorter operation time, earlier return to normal diet, less pain on 1st to 3rd day postoperatively.
Adolescent ; Child ; Child, Preschool ; Cold Temperature ; Cryosurgery ; methods ; Dissection ; methods ; Female ; Humans ; Male ; Tonsillectomy ; methods ; Tonsillitis ; surgery ; Treatment Outcome

The treatment of acute necrotizing fasciitis consists of early and aggressive incision and drainage, together with effective antibiotics and active organ support. On January 4th, 2018, a 50-year-old man admitted to intensive care unit (ICU) of Peking University Third Hospital was diagnosed with acute necrotizing fasciitis in the right lower extremity caused by group A Streptococcal infection complicated with multiple organ dysfunction. The disease progressed rapidly. The lesion rapidly spread from the right calf to the entire right lower limb and was associated with septic shock and multiple organ dysfunction within a few hours after admission. The treatment included surgical debridement and amputation of his right lower extremity, multiple antimicrobials combination therapy, gamma globulin-assisted anti-infection, anti-shock, bedside continuous hemofiltration and other comprehensive treatment. The patient was successfully cured. The key to successful treatment of this patient is that when the acute necrotizing fasciitis progresses rapidly, amputation should be done to debride the infected lesions, together with early empirical broad-spectrum antibiotics combination therapy and active organ function support treatment.

Objective:Multidrug-resistant tuberculosis(MDR-TB)has a high mortality and is always one of the major challenges in global TB prevention and control.Analyzing the factors that may impact the adverse outcomes of MDR-TB patients is helpful for improving the systematic management and optimizing the treatment strategies for MDR-TB patients.For follow-up data,the Cox proportional hazards regression model is an important multifactor analysis method.However,the method has significant limitations in its application,such as the fact that it is difficult to deal with the impacts of small sample sizes and other practical issues on the model.Therefore,Bayesian and conventional Cox regression models were both used in this study to analyze the influencing factors of death in MDR-TB patients during the anti-TB therapy,and compare the differences between these 2 methods in their application. Methods:Data were obtained from 388 MDR-TB patients treated at Lanzhou Pulmonary Hospital from November 1,2017 to March 31,2021.Survival analysis was employed to analyze the death of MDR-TB patients during the therapy and its influencing factors.Conventional and Bayesian Cox regression models were established to estimate the hazard ratios(HR)and their 95% confidence interval(95% CI)for the factors affecting the death of MDR-TB patients.The reliability of parameter estimation in these 2 models was assessed by comparing the parameter standard deviation and 95% CI of each variable.The smaller parameter standard deviation and narrower 95% CI range indicated the more reliable parameter estimation. Results:The median survival time(1st quartile,3rd quartile)of the 388 MDR-TB patients included in the study was 10.18(4.26,18.13)months,with the longest survival time of 31.90 months.Among these patients,a total of 12 individuals died of MDR-TB and the mortality was 3.1%.The median survival time(1st quartile,3rd quartile)for the deceased patients was 4.78(2.63,6.93)months.The majority of deceased patients,accounting for 50%,experienced death within the first 5 months of anti-TB therapy,with the last mortality case occurring within the 13th month of therapy.The results of the conventional Cox regression model showed that the risk of death in MDR-TB patients with comorbidities was approximately 6.96 times higher than that of patients without complications(HR=6.96,95% CI 2.00 to 24.24,P=0.002)and patients who received regular follow-up had a decrease in the risk of death by approximately 81% compared to those who did not receive regular follow-up(HR=0.19,95% CI 0.05 to 0.77,P=0.020).In the results of Bayesian Cox regression model,the iterative history plot and Blue/Green/Red(BGR)plot for each parameter showed the good model convergence,and parameter estimation indicated that the risk of death in patients with a positive first sputum culture was lower than that of patients with a negative first sputum culture(HR=0.33,95% CI 0.08 to 0.87).Additionally,compared to patients without complications,those with comorbidities had an approximately 6.80-fold increase in the risk of death(HR=7.80,95% CI 1.90 to 21.91).Patients who received regular follow-up had a 90% reduction in the risk of death compared to those who did not receive regular follow-up(HR=0.10,95% CI 0.01 to 0.30).The comparison between these 2 models showed that the parameter standard deviations and corresponding 95% CI ranges of other variables in the Bayesian Cox model were significantly smaller than those in the conventional model,except for parameter standard deviations of receiving regular follow-up(Bayesian model was 0.77;conventional model was 0.72)and pulmonary cavities(Bayesian model was 0.73;conventional model was 0.73). Conclusion:The first year of anti-TB therapy is a high-risk period for mortality in MDR-TB patients.Complications are the main risk factors of death in MDR-TB patients,while patients who received regular follow-up and had positive first sputum culture presented a lower risk of death.For data with a small sample size and low incidence of outcome,the Bayesian Cox regression model provides more reliable parameter estimation than the conventional Cox model.