AIM:To study the influence of material and configuration of microfiltration membranes on preparation of mono-ammonium glycyrrhizinate liposome(MGL) by membrane extrusion. METHODS: MGL were preparated by conventional rotary-evaporated film dispersion method and sonication,and MGL were extruding through 9 kinds of microfiltration membranes with 3 kinds of aperture specs of 0.1,0.2,0.45 ?m.The methods were evaluated by the filtration efficiency and the leakage rate of drug from liposomes. RESULTS: MGL were easily extruding through microfiltration membranes of NY6,PTFE,PVDF,PP and PC.MGL could not extruding through microfiltration membranes of PES,CN,CA and CA-CN.MGL become hardly to extrude through microfiltration membranes when increasing in the concentration of liposomes or the content of cholesterol.The leakage rate of drug from MGL was small when extruding through microfiltration membranes. CONCLUSION: The material and configuration of microfiltration membranes can remarkably influence the preparation of MGL by membrane extrusion method,and PC microfiltration membrane was selected.

Objective To investigate the anticancer activities of resveratrol on malignant melanoma cells in vitro and involved mechanisms. Methods A375 human malignant melanoma cells and B16-F1mouse malignant melanoma cells were cultured and treated with various concentrations of resveratrol for different durations. The cell proliferation, apoptosis and cycle of both B16-F1 and A375 cells were detected with MTT assay, Annexin V-F1TC/propidium iodide (PI) double staining flow cytometry and propidium iodide flow cytometry, respectively. Western blot analysis was performed to measure the expression of Bcl-2 and Bax protein in both cells. Results Resveratroi inhibited the proliferation of A375 and B16-F1 cells in a time- and dose- dependent manner. The apoptosis rate of A375 cells was (16.7±2.1 )%, (17.2±1.7)% and (52.3±4.1 )% after treatment with resveratrol of 25 μmol/L for 24 hours, resveratrol of 100 μmol/L for 12 and 72 hours, respectively;, and resveratrol of 100 μmol/L induced the apoptosis of B16-F1 at a rate of ( 18.4±1.6)%, (39.6±3.3 )% and (56.7±4.5 )% at 12, 24 and 72 hours, respectively. Flow cytometry showed that A375 and B16-F1 cells treated with resveratrol were arrested in the G1 phase of cell cycle, and the blocking effect increased in a dose-dependent manner. The percentage of A375 and B16-F1 cells in G1 phase was (40.51±3.97 )% and (41.34±3.12 )%, respectively, after 24-hour treatment with resveratrol of 25 μmol/L,(55.64±4.95)% and (53.93±5.12)%, respectively with resveratrol of 100μmol/L for the same duration.The expression of Bcl-2 protein was decreased in malignant melanoma cells treated with resveratrol,while that of Bax protein increased. Conclusions Resveratrol can effectively inhibit the proliferation of malignant melanoma cells by regulating the cell cycle and inducing cell apoptosis, which seems to be associated with the regulation of Bcl-2 and Bax expressions.

The histological and immunohistological features of two cases of cutaneous lymphadenoma was studied. A single, erythematous nodule with smooth surface developed on the face of both patients. The lesion slowly progressed. Histology revealed irregular epithelial lobules in the dermis which showed a peripheral palisaded border of basaioid-like cells as well as a center composed of clear cells. Some epithelial lobules and surrounding stroma were infiltrated by numerous small lymphocytes. Immunohistological study showed that the lymphocytic infiltration was predominantly composed of T cells (CD3 positive) along with a small number of B cells (CD20 positive). Within epithelial lobules and surrounded stroma, there were numerous dendritic cells that were positive for S-100 and CDia but negative for cytokeratin 7, cytokeratin 20 or carcino-embryonic antigen. In the center of epithelial lobules in one case, a few cells positive for epithe-lial membrane antigen and CD30 was observed. The diagnosis of cutaneous lymphadenoma was made according to the pathological and immunohistochemical findings, and the infiltration was predominated by CD3-positive lymphocytes in this uncommon epithelial neoplasm.

Objective To compare the reactive oxygen species (ROS) expression in human neutrophils phagocytizing Sporothrix Schenckii and Candida albicans yeast cells,and to compare the fungicidal activity of human neutrophils against Sporothrix Schenckii and Candida albicans.Methods Human neutrophils were isolated from peripheral blood by using density gradient centrifugation method,and cultured with the presence of the yeast phase of a Sporothrix Schenckii clinical isolate and a standard strain of Candida albicans (ATCC 90028)at a multiplicity of infection of 10 or 1 for 60-210 minutes.Subsequently,flow cytometry with ROS probe (2',7'-dichlorofluorescein diacetate,DCFH-DA) was carried out to for the real time detection of intracellular ROS level,confocal laser scanning microscopy (CLSM) for the observation of ROS distribution.In addition,the fungicidal efficiency of neutrophils against Sporothrix Schenckii and Candida albicans was estimated by the number of colonies after additional culture of neutrophil lysates on brain-heart infusion agar (BHIA) medium.Statistical analysis was done by using univariate analysis of variance and LSD-t test.Results The intracellular ROS level peaked at 60 minutes in neutrophils incubated with Sporothrix Schenckii yeast cells,then decreased rapidly from 60 minutes to 210 minutes.Compared with the neutrophils incubated with Candida albicans yeast cells,those with Sporothrix Schenckii yeast cells showed a higher ROS level (expressed as mean fluorescence intensity) at 60minutes (159.67 ± 11.34 vs.112.22 ± 9.66,P＜ 0.01),but a lower ROS level at 120 minutes (89.01 ± 9.81 vs.110.25 ± 7.28,P＜ 0.05) and 180 minutes (57.63 ± 8.46 vs.109.98 ± 9.00,P＜ 0.01).CLSM revealed that ROS was mainly distributed in neutrophils with phagocytized fungal spores,and especially on the surface of phagocytized spores.Furthermore,the percentage of yeast cells killed by neutrophils was significantly lower for Sporothrix Schenckii than for Candida albicans at 180 minutes (19.21％ ± 3.68％ vs.26.63％ ± 4.97％,P ＜ 0.01).Conclusions Differential expression of intracellular ROS was observed in neutrophils after phagocytosis of Candida albicans and Sporothrix schenckii.Neutrophils exert a stronger fungicidal activity against Sporothrix Schenckii in comparison with Candida albicans,which may be associated with the rapid decrease of ROS level in neutrophils after phagocytosis.

Objective To estimate the value of BIOMED-2 primers for the detection of T cell receptor γ (TCR-γ) gene rearrangements in different types of specimens from patients with mycosis fungoides (MF).Methods Totally,15 paraffin-embedded tissue specimens,14 fresh tissue specimens and 18 whole blood specimens were obtained from 28 patients with MF,and subjected to DNA extraction.BIOMED-2 multiplex PCR tubes TCRγ (A+B) were used for the analysis of TCRγgene rearrangements.Data were processed by SPSS 13.0 software,and statistical analysis was done by chi-square test and Fisher's exact probability test.Results TCR-γ gene rearrangements were detected in 3 paraffin-embedded tissue specimens,11 fresh tissue specimens and 12 blood specimens,with significant differences in the detection rate between the three samples (x2 =13.047,P ＜ 0.01).The fresh tissue samples showed a significantly higher detection rate than the paraffin-embedded tissue samples (X2 =12.523,P ＜ 0.01).The detection rate of TCRγgene rearrangements was 3/6 in paraffin-embedded tissue samples collected in 2011,significantly higher than that in the other 9 paraffin-embedded tissue samples collected before 2011 (Fisher's exact probability test,P =0.044),but similar to that in 14 fresh tissue specimens (12/14,Fisher's exact probability test,P =0.044).Decreased detection rate of TCRγ gene rearrangements was observed in blood samples compared with fresh tissue specimens,but no statistical difference was observed between the two types of specimens (x2 =2.358,P ＞ 0.05).Conclusions BIOMED-2 multiplex PCR tubes TCRγ(A+B) are suitable for the detection of clonal rearrangements of TCRγgene in different types of specimens,especially in fresh tissue specimens,from patients with MF.

ObjectiveTo study the correlation between the etiology and clinical features of erythroderma.MethodsThe clinical data on 182 patients with erythroderma were retrospectively collected and analyzed.ResultsThe male-to-female ratio was 2.8 ∶ 1 and the average age at onset was 58.6 ± 14.6 years.Of the 182 cases,135 (74.2％) were due to pre-existing dermatoses,14 (7.7％) to drug reaction,8 (4.4％) to malignancies,while 25(13.7％) had no obvious precipitating factors.The most frequent triggering factor was systemic consumption of drugs(52 patients,28.6％ ),and glucocorticosteroid was the most prevalent causative drug.Seventy-six patients were followed up,recurrence was observed in 14 patients but not in 58 patients,and 5 patients died,2 patients with idiopathic erythroderma were finally diagnosed with mycosis fungoides (MF)after multiple skin biopsies.ConclusionsPre-existing dermatoses are the most frequent cause of erythroderma.Idiopathic erythroderma is liable to relapse,possibly associated with malignancies,and should be closely followed up.

Objective To examine the proportion and reasons of drop-out from antiretroviral therapy(ART)among 8 367 adult HIV-infected individuals in Dehong prefecture,Yunnan province. Methods All adult HIV-infected patients receiving ART before September 30 of 2014 were examined for the situation of drop-out from ART. Results The proportion of drop-out from ART among adult HIV-infected patients in Dehong prefecture was 14.4%(1 202/8 367). Results from the univariate logistic regression analyses indicated that drop-out from ART was significantly correlated with factors as:living area,gender,age,marital status,HIV transmission route,baseline CD4+T cell counts and initial treatment regimen of the patients. After adjusted for potential confounding variables by multiple logistic regression model,drop-out from ART was significantly correlated with residential area,marital status,HIV transmission route,baseline CD4+T cell count and initial treatment regimen of the patients. HIV-infected patients who were living in Mangshi city,Lianghe county or Yingjiang county,being married or living with partner,HIV infection through sexual contact,with baseline CD4+T cell counts≤200 cells/mm3,and ART included in the initial treatment regimen etc.,were less likely to drop out from ART. The proportion of drop out from ART was significantly decreasing along with the increasing time of ART. Data from specific investigation revealed that among the 1 202 patients who dropped out from ART,704(58.6%)were lost to follow-up,303(25.2%) did not adhere to treatment,74(6.2%)moved out the region,64(5.3%)were Burmese that had returned to Burma,29 (2.4%) stopped the treatment according to doctors’advice,18(1.5%)were incarcerated and 10 (0.8%) were under other reasons. Reasons for the drop-out varied,according to the situation of patients. Conclusion The proportion of drop-out from ART varied significantly according to the characteristics of HIV-infected patients in Dehong prefecture that underscoring the needs for tailored responses to reduce drop-out of ART. Focus should be targeted on reducing the loss to follow-up and improving the treatment adherence.

Objective To examine the proportion and reasons of drop-out from antiretroviral therapy(ART)among 8 367 adult HIV-infected individuals in Dehong prefecture,Yunnan province. Methods All adult HIV-infected patients receiving ART before September 30 of 2014 were examined for the situation of drop-out from ART. Results The proportion of drop-out from ART among adult HIV-infected patients in Dehong prefecture was 14.4%(1 202/8 367). Results from the univariate logistic regression analyses indicated that drop-out from ART was significantly correlated with factors as:living area,gender,age,marital status,HIV transmission route,baseline CD4+T cell counts and initial treatment regimen of the patients. After adjusted for potential confounding variables by multiple logistic regression model,drop-out from ART was significantly correlated with residential area,marital status,HIV transmission route,baseline CD4+T cell count and initial treatment regimen of the patients. HIV-infected patients who were living in Mangshi city,Lianghe county or Yingjiang county,being married or living with partner,HIV infection through sexual contact,with baseline CD4+T cell counts≤200 cells/mm3,and ART included in the initial treatment regimen etc.,were less likely to drop out from ART. The proportion of drop out from ART was significantly decreasing along with the increasing time of ART. Data from specific investigation revealed that among the 1 202 patients who dropped out from ART,704(58.6%)were lost to follow-up,303(25.2%) did not adhere to treatment,74(6.2%)moved out the region,64(5.3%)were Burmese that had returned to Burma,29 (2.4%) stopped the treatment according to doctors’advice,18(1.5%)were incarcerated and 10 (0.8%) were under other reasons. Reasons for the drop-out varied,according to the situation of patients. Conclusion The proportion of drop-out from ART varied significantly according to the characteristics of HIV-infected patients in Dehong prefecture that underscoring the needs for tailored responses to reduce drop-out of ART. Focus should be targeted on reducing the loss to follow-up and improving the treatment adherence.

Objective:To investigate and analysis of the occurrence and influencing factors of sarcopenia in elderly critically ill patients in the intensive care unit (ICU).Methods:A prospective cohort study was conducted. Elderly patients (aged ≥ 60 years) admitted to the ICU of Tianjin Medical University General Hospital from November 2023 to June 2024 were enrolled. Clinical records were collected, and conduct muscle mass and strength measurements, as well as upper arm circumference and calf circumference were measured. Appendicular skeletal muscle index (ASMI) of less than 7.0 kg/m 2 for males and less than 5.7 kg/m 2 for females was defined as reduced muscle mass, grip strength of less than 28 kg for males and less than 18 kg for females was defined as decreased muscle strength, patients meeting both low muscle mass and low muscle strength criteria were diagnosed with sarcopenia. According to the diagnostic criteria for sarcopenia, patients were divided into sarcopenia group and non-sarcopenia group. Multivariate Logistic regression analysis was applied to identify risk factors for sarcopenia in the elderly and to develop a predictive model for the occurrence of sarcopenia. The predictive value of various risk factors for sarcopenia in elderly critically ill patients were evaluated by receiver operator characteristic curve (ROC curve). The Kaplan-Meier curve for the length of ICU stay of two groups patients were drawn. Results:Finally, 540 elderly critically ill patients were included, including 43 patients with sarcopenia, and the incidence of sarcopenia was 8.0%. Univariate analysis showed that there were significantly differences in body mass index (BMI), number of hospitalizations in the past year, the length of ICU stay, ventilation mode, duration of mechanical ventilation, pre-admission exercise habits, nutritional support methods, upper arm circumference, calf circumference, and albumin infusion between the sarcopenia group and the non-sarcopenia group. Multivariate Logistic regression analysis showed that BMI [odds ratio ( OR) = 0.79, 95% confidence interval (95% CI) was 0.67-0.93, P = 0.004], calf circumference ( OR = 0.64, 95% CI was 0.54-0.76, P < 0.001), and duration of mechanical ventilation ( OR = 1.06, 95% CI was 1.01-1.12, P = 0.034) were associated with an increased risk of sarcopenia in elderly critically ill patients. The ROC curve results showed that the area under the curve (AUC) and 95% CI of BMI, calf circumference, and duration of mechanical ventilation for predicting sarcopenia in elderly critically ill patients were 0.828 (0.767-0.888), 0.889 (0.844-0.933), and 0.397 (0.299-0.496), respectively, with cut-off values of 22.95 kg/m 2, 28.25 cm, and 50.50 days, respectively. The Kaplan-Meier curve showed that the cumulative survival rate of patients with sarcopenia was significantly lower than that of the non-sarcopenia group (Log-Rank test: χ 2 = 5.619, P = 0.018). Conclusion:Lower BMI, smaller calf circumference, and longer duration of mechanical ventilation are associated with an increased risk of sarcopenia in critically ill elderly patients.

OBJECTIVETo examine the proportion and reasons of drop-out from antiretroviral therapy (ART) among 8 367 adult HIV-infected individuals in Dehong prefecture, Yunnan province.

METHODSAll adult HIV-infected patients receiving ART before September 30 of 2014 were examined for the situation of drop-out from ART.

RESULTSThe proportion of drop-out from ART among adult HIV-infected patients in Dehong prefecture was 14.4% (1 202/8 367). Results from the univariate logistic regression analyses indicated that drop-out from ART was significantly correlated with factors as: living area, gender, age, marital status, HIV transmission route, baseline CD4⁺ T cell counts and initial treatment regimen of the patients. After adjusted for potential confounding variables by multiple logistic regression model, drop-out from ART was significantly correlated with residential area, marital status, HIV transmission route, baseline CD4⁺ T cell count and initial treatment regimen of the patients. HIV-infected patients who were living in Mangshi city, Lianghe county or Yingjiang County, being married or living with partner, HIV infection through sexual contact, with baseline CD4⁺ T cell counts ≤ 200 cells/mm³, and ART included in the initial treatment regimen etc., were less likely to drop out from ART. The proportion of drop out from ART was significantly decreasing along with the increasing time of ART. Data from specific investigation revealed that among the 1 202 patients who dropped out from ART, 704 (58.6%) were lost to follow-up, 303 (25.2%) did not adhere to treatment, 74 (6.2%) moved out the region, 64 (5.3%) were Burmese that had returned to Burma, 29 (2.4%) stopped the treatment according to doctors' advice, 18 (1.5%) were incarcerated and 10 (0.8%) were under other reasons. Reasons for the drop-out varied, according to the situation of patients.

CONCLUSIONThe proportion of drop-out from ART varied significantly according to the characteristics of HIV-infected patients in Dehong prefecture that underscoring the needs for tailored responses to reduce drop-out of ART. Focus should be targeted on reducing the loss to follow-up and improving the treatment adherence.