Phage antibody library technology in tumor therapy: a review / 生物工程学报

Tumor is a serious threat to human health. At present, surgical resection, chemoradiotherapy, targeted therapy and immunotherapy are the main therapeutic strategies. Monoclonal antibody has gradually become an indispensable drug type in the clinical treatment of cancer due to its high efficiency and low toxicity. Phage antibody library technology (PALT) is a novel monoclonal antibody preparation technique. The recombinant immunoglobulin variable region of heavy chain (VH)/variable region of light chain (VL) gene is integrated into the phage vector, and the antibody is expressed on the phage surface in the form of fusion protein to obtain a diverse antibody library. Through the process of adsorption-elution-amplification, the antibody library can be screened to obtain the antibody molecule with specific binding antigen as well as its gene sequence. PALT has the advantages of short antibody production cycle, strong plasticity of antibody structure, large antibody yield, high diversity and direct production of humanized antibodies. It has been used in screening tumor markers and preparation of antibody drugs for breast cancer, gastric cancer, lung cancer and liver cancer. This article reviews the recent progress and the application of PALT in tumor therapy.

Inhibitory and clearance effect of azithromycin combined with levofloxacin on biofilm of different ST types of Klebsiella pneumoniae / 中国临床药理学与治疗学

AIM: Given the biofilm formation abili- ty of different ST-type Klebsiella pneumoniae, our study was aimed at exploring the inhibitory and clearance of azithromycin combined with levofloxacin on the biofilm of Klebsiella pneumoniae of different ST-types and providing a new strategy for the prevention of biofilm formation in the treatment of post-infectious Klebsiella pneumoniae. METHODS: 9 strains of Klebsiella pneumoniae from all susceptibility groups, 19 strains of Klebsiella pneumoniae producing extended-spectrum β - lactamases (ESBLs), and 37 strains of Carbapenem-resistant Klebsiella pneumoniae (CRKP) were randomly collected from the samples of patients hospitalized in the First Affiliated Hospital of Wannan Medical College from August 2019 to November 2021. The isolates were identified using VITEK MS IVD KB V3.2 and VITEK 2-Compact 60. Multilocus sequence typing (MLST) was performed to analyze the homology of each strain; crystal violet staining was used for semi -quantitative detection of biofilm to compare the differences in biofilm formation ability between different ST-type Klebsiella pneumoniae. Different ST-type strains were selected, and the partial inhibitory concentration index (FICI) was calculated by micro broth dilution method to judge the combination effect and select the optimal combination concentration; crystalline violet staining method was used to investigate the inhibition and clearance effect of azithromycin combined with levofloxacin on the biofilm of different ST-type Klebsiella pneumoniae; laser scanning confocal fluorescence microscopy was used to observe the structural changes of the biofilm of Klebsiella pneumoniae before and after the effect of the antibacterial drugs. RESULTS: MLST typing results showed that the sensitive group of Klebsiella pneumoniae strains had 8 sequences such as ST86, ST727, etc., the ESBLs group strains belonged to 14 sequence types such as ST15, ST37, ST11, etc., of which ST15 accounted for 26.32% (5 / 19). The CRKP group strains belonged to 9 sequence types such as ST11, ST15, ST656, etc., of which ST11 accounted for 48.65% (18/37), ST15 accounted for 27.03% (10/37); ST15 (ESBLs), ST11 (CRKP), ST15 (CRKP) type Klebsiella pneumoniae biofilms all reached maturity on the 5th day, the ST15 (ESBLs) group had a stronger ability to produce material to be membranous than the ST15 (CRKP) group. The ST11 (CRKP) group had a stronger ability to produce material to be membranous than the ST15 (CRKP) group (Pazithromycin>levofloxacin, and the highest clearance rate was 44.79%. CONCLUSION: There are differences in biofilm formation ability between different ST-type Klebsiella pneumoniae, and azithromycin combined with levofloxacin has a better inhibitory effect on different ST-type Klebsiella pneumoniae biofilm, conbined application can be used in the treatment of biofilm infections early stage.

Biomarkers for early screening and diagnosis of breast cancer: a review / 生物工程学报

The estimated new cases of breast cancer (BC) patients were 2.26 million in 2020, which accounted for 11.7% of all cancer patients, making it the most prevalent cancer worldwide. Early detection, diagnosis and treatment are crucial to reduce the mortality, and improve the prognosis of BC patients. Despite the widespread use of mammography screening as a tool for BC screening, the false positive, radiation, and overdiagnosis are still pressing issues that need to be addressed. Therefore, it is urgent to develop accessible, stable, and reliable biomarkers for non-invasive screening and diagnosis of BC. Recent studies indicated that the circulating tumor cell DNA (ctDNA), carcinoembryonic antigen (CEA), carbohydrate antigen 15-3 (CA15-3), extracellular vesicles (EV), circulating miRNAs and BRCA gene from blood, and the phospholipid, miRNAs, hypnone and hexadecane from urine, nipple aspirate fluid (NAF) and volatile organic compounds (VOCs) in exhaled gas were closely related to the early screening and diagnosis of BC. This review summarizes the advances of the above biomarkers in the early screening and diagnosis of BC.

Advances in predictive biomarkers associated with immune checkpoint inhibitors for tumor therapy / 生物工程学报

Malignant tumors are diseases that seriously threaten human health and social development. Traditional tumor therapies such as surgery, radiotherapy, chemotherapy and targeted therapy cannot fully meet the needs of clinical treatment, and emerging immunotherapy has become a research hotspot in the field of tumor treatment. Immune checkpoint inhibitors (ICIs) have been approved as a tumor immunotherapy method for the treatment of various tumors, such as lung cancer, liver cancer, stomach cancer and colorectal cancer, etc. However, during the clinical use of ICIs, only a small number of patients experienced durable responses, which also led to drug resistance and adverse reactions. Therefore, the identification and development of predictive biomarkers is crucial to improve the therapeutic efficacy of ICIs. The predictive biomarkers of tumor ICIs mainly include tumor biomarkers, tumor microenvironment biomarkers, circulation-related biomarkers, host environmental biomarkers and combinatorial biomarkers. They are of great significance for screening, individualized treatment and prognosis evaluation of tumor patients. This article reviews the advances of predictive markers for tumor ICIs therapy.

Distribution of Traditional Chinese Medicine Patterns and Analysis of Factors Related to Acute Exacerbation in Group E of Chronic Obstructive Pulmonary Disease / 中医杂志

ObjectiveTo analyze the distribution of traditional Chinese medicine （TCM） patterns as well as factors related to acute exacerbation in group E of chronic obstructive pulmonary disease （COPD）. MethodsThe general data of 161 COPD patients， including gender， age， body mass index （BMI）， disease course， smoking history， and past history， were collected. In terms of the four examinations of TCM， the differentiated patterns included phlegm-heat obstructing the lung， turbid phlegm obstructing the lung， phlegm stasis obstructing the lung， lung-spleen qi deficiency， and lung-kidney deficiency. The modified British Medical Research Council （mMRC） scale and COPD assessment test （CAT）， the pulmonary function indicators including forced expiratory volume in the first second （FEV1） and ratio of forced expiratory volume to forced vital capacity at second 1 （FEV1/FVC）， GOLD grade， and the patient's acute exacerbations in the previous year were recorded. Multivariate regression analysis was performed using logistic regression model to determine the relevant factors of patients in COPD group E. The distribution of acute exacerbations in different TCM symptom patients in group E was analyzed. ResultsThere were 80 patients （49.69%） in group E and 81 patients （50.31%） in non-group E. In group E， 23 （28.75%） patients had a history of two acute exacerbations， while 35 （43.75%） had three acute exacerbations， and 22 （27.5%） had more than three acute exacerbations. There were 13 （16.25%） cases of phlegm-heat obstructing the lung pattern， 6 （7.5%） cases of turbid phlegm obstructing the lung pattern， 8 （10%） cases of phlegm stasis obstructing the lung pattern， 22 cases （27.5%） of lung-spleen qi deficiency pattern， and 31 （38.75%） cases of lung-kidney deficiency pattern. There were significant differences in smoking history， disease course， TCM pattern， TCM syndrome score， mMRC score， and CAT score between groups （P＜0.05）. A total of 107 of the 161 patients completed pulmonary function tests， and the differences in FEV1， FEV1/FVC and GOLD grades between groups were statistically significant （P＜0.05）. Multivariate regression analysis showed that TCM pattern， TCM syndrome score and CAT score were statistically significant factors for COPD patients in group E （P＜0.05）. There were statistically significant differences in the number of acute exacerbations in different TCM patterns in group E （P＜0.05）. The patients with two acute exacerbations in the past year were mainly phlegm-heat obstructing the lung and lung-spleen qi deficiency patterns， while the three acute exacerbations were mainly seen in lung-spleen qi deficiency and lung-kidney deficiency patterns， and more than three exacerbations were more common with lung -kidney deficiency pattern. ConclusionsPatients in COPD group E were mainly the lung-spleen qi deficiency and lung-kidney deficiency patterns. Deficiency of healthy qi is the main reason for the increase in the number of acute exacerbations， and TCM patterns and CAT score were the main related factors.

Expression of IL-2/IL-15Rβ on memory CD3 + CD8 + CD45RO + T cells in chronic hepatitis B patients undergoing antiviral treatment and its significance / 中华临床感染病杂志

Objective:To investigate the expression of IL-2/IL-15 receptor β subunit (IL-2/IL-15Rβ) on memory CD3 + CD8 + CD45RO + T cells in patients with chronic hepatitis B (CHB) receiving antiviral treatment and its significance. Methods:Sixty-eight patients with chronic active hepatitis B (CAHB) and 47 asymptomatic hepatitis B virus (HBV) carriers attending in the Department of Infectious Diseases, the First Affiliated Hospital of Wannan Medical College from March 2019 to December 2020 were enrolled in the study; and 30 health subjects were also enrolled as healthy control group. Among 60 CAHB patients there were 30 cases with positive HBeAg and 30 cases with negative HBeAg. All CAHB patients received nucleos(t)ide analogue therapy, the HBV-related markers, Alanine aminotransferase (ALT) and the expression of IL-2/IL-15Rβ on CD3 + CD8 + CD45RO + T cells were determined and compared between HBeAg-positive and negative patients, before and after treatment. Normal distribution measurement data among 3 groups were compared with One-way ANOVA; normal distribution measurement data between 2 groups were compared with paired samples t test; non-normal distribution measurement data between the two groups were compared with Mann-Whitney U test; Pearson’s correlation coefficient was performed for correlation analysis. P<0.05 was considered statistically significant. Results:The proportion of CD8 + CD45RO + T cells on PBMC CD3 + T cells in CAHB group [(8.6±3.7)%] was higher than that of asymptomatic HBV carriers group [(5.7±2.5)%] and healthy control group [(5.5±1.5)%] (all P<0.05). The expression percentage of IL-2/IL-15Rβ on PBMC CD3 + CD8 + CD45RO + T cells in CAHB group [(6.8±4.7)%] was higher than that of asymptomatic HBV carriers group [(4.7±2.8)%] and healthy control group [(4.3±2.2)%] (all P<0.05). The MFI of IL-2/IL-15Rβ on PBMC CD3 + CD8 + CD45RO + T cells in CAHB group (243±168) was higher than those of asymptomatic HBV carriers group (160±91) and healthy control group [160±63] (all P<0.05). The expression percentage and MFI of IL-2/IL-15Rβ on PBMC CD3 + CD8 + CD45RO + T cells were positively correlated with the percentage of CD3 + CD8 + CD45RO + T cells in CAHB patients ( r=0.33 and 0.28, all P<0.05). The proliferation percentage of PBMC CD3 + CD8 + CD45RO + T cells in CAHB group[ (43.7±16.0)%] was higher than that of asymptomatic HBV carriers group [(29.1±9.4)%] and healthy control group [(26.8±9.6)%] after stimulation with Anti-CD3+ super-2 (all P<0.05). After the expression of IL-2/IL-15Rβ was blocked, the proliferation percentage of CD3 + CD8 + CD45RO + T cells was decreased [(11.2±6.3)%] compared with the untreated CAHB group ( P<0.05). The percentages of PBMC CD3 + CD8 + CD45RO + T cells secreting IFN-γ, IL-2 and TNF-α in CAHB group were (13.8±5.4)%, (14.0±4.3)% and (12.3±4.6)% respectively, which were higher than those of asymptomatic HBV carriers [(8.4±2.6)%, (9.4±3.2)% and (6.8±3.3)%] and healthy control group [(6.9±2.7)%, (9.9±3.0)% and (7.7±3.8)%] after stimulation with Anti-CD3+ super-2 (all P<0.05). After the expression of IL-2/IL-15Rβ was blocked, the percentages of PBMC CD3 + CD8 + CD45RO + T cells secreting IFN-γ [(2.4±1.6)%], IL-2 [(4.1±1.9)%] and TNF-α [(4.1±1.8)%] were decreased compared with the untreated CAHB group (all P<0.05). HBeAg, ALT, the expression percentage and MFI of IL-2/IL-15Rβ on CD3 + CD8 + CD45RO + T cells were 521.4 (68.9, 1 339.0) COI, 292 (160, 528) U/L, (6.4±3.2)% and (239±136) in 30 HBeAg-positive CAHB patients before treatment, which were higher than those after treatment [3.5(1.5, 17.5)COI、20(14, 31) U/L, (4.1±2.4)% and (134±58)] ( Z=5.337 and 6.403, t=3.229 and 3.892, all P<0.05). HBsAg, ALT, the expression percentage and MFI of IL-2/IL-15Rβ on CD3 + CD8 + CD45RO + T cells were (5 310±2 851) COI, (328±207) U/L, (7.1±5.8)% and (252±110) in 30 HBeAg-negative CAHB patients before treatment, which were higher than those after 48 weeks of treatment [(3 811±2 495) COI, (33±14) U/L, (4.6±2.9)% and (154±73)] ( t=2.167, 5.595, 2.116 and 2.383, all P<0.05). Conclusion:The study suggests that up-regulated expression of IL-2/IL-15Rβ is associated with elevated frequency, proliferation and secretion function of memory CD3 + CD8 + CD45RO + T cells in CAHB patients.

Development of a reference substance for live bacterial count of Streptococcicosis live vaccines / 生物工程学报

This study attempts to develop a reference substance for the live bacteria count of Streptococcicosis live vaccines in order to evaluate the validity of live bacterial count in inspection and testing. We prepared a batch of live Streptococcus suis reference substance for live bacterial count, tested their physical property, purity, vacuum degree, remaining moisture, and determined their homogeneity, thermal stability and transportation stability. Moreover, we organized collaborative calibration to assign count values to the reference substance and determine the shelf life of the reference substance in 12 months. The results showed that the physical property, the purity, the remaining moisture and the vacuum degree of the reference substance were all in compliance with the requirements of the Chinese Veterinary Pharmacopoeia. The homogeneity test showed that the coefficient of variation of the count of the reference substance was less than 10%, indicating a good homogeneity. Transportation stability test showed that the reference substance remained active after 72 h transportation in summer and winter with the package of styrofoam boxes and ice packs. Thermal stability test showed that the reference substance could be stored for up to 3 months at -20 °C, or up to 21 days at 4 °C. According to the collaborative calibration, the reference vaccine was assigned a count value range of (8.5-12.1)×107 CFU/ampoule. The shelf life test showed that the reference substance was stable for 12 months when stored at -70 °C. The reference substance could provide a reference for the live bacterial count of Streptococcicosis live vaccines. Moreover, it could also be used as a reference to evaluate the quality of corresponding agar media.

Expression of MCM4/PCNA in gastric cancer and its potential as a potential biomarker / 国际生物医学工程杂志

Objective:To analyze the expressions of minichromosome maintenance protein 4(MCM4) and proliferating cell nuclear antigen (PCNA) in gastric cancer tissues, to explore the relationship between MCM4/PCNA and gastric cancer, and to investigate the possibility of MCM4/PCNA as a potential biomarker for gastric cancer.Methods:Bioinformatics methods were used to analyze the mRNA expressions of MCM4 and PCNA in gastric cancer tissues and adjacent normal tissues. The clinicopathological data of 69 patients with gastric cancer who underwent surgery were retrospectively analyzed. The expression levels of MCM4 and PCNA in gastric cancer tissues and adjacent normal tissues were detected by immunohistochemistry, and their relationship with the clinicopathological characteristics of gastric cancer patients was analyzed.Results:The mRNA levels of MCM4 and PCNA in gastric cancer tissues are significantly higher than those in adjacent normal tissues (all P<0.05). The expression of MCM4 is correlated with the tumor size of gastric cancer ( P=0.037), but there is no significant correlation with gender, age and tumor grade (all P>0.05). Both MCM4 and PCNA proteins are highly expressed in gastric cancer patients. Conclusions:The expression levels of MCM4 and PCNA have a clear correlation with the occurrence of gastric cancer. MCM4 and PCNA are expected to be potential biomarkers for the diagnosis and treatment of gastric cancer.

Inhibitory and scavenging ability of ambroxol combined with levofloxacin on the bacterial biofilm of Klebsiella pneumoniae / 中国临床药理学与治疗学

AIM: To study the inhibitory and scavenging effects of ambroxol combined with levofloxacin on the bacterial biofilm of Klebsiella pneumoniae, and to provide a new strategy to treat and antagonize the formation of the biofilm. METHODS: We collected Klebsiella pneumoniae of different resistance and divided them into sensitive group (wild bacteria group), ESBLs group and CRKP group with 15 strains in each group and performed biological semi-quantitative detection of its biofilm by crystalline violet staining method. After selecting 3 strains with similar membrane yields from each group, we determined the minimum inhibitory concentration (MIC) of ambroxol and levofloxacin against Klebsiella pneumoniae by micro broth dilution method.In addition, we determined the effects of ambroxol in different concentrations on the MIC of levfloxacin by the micrdilution checkerboard techniques and calculated the partial inhibitory concentration index (FIC) to determine the joint effect and select the best synergistic concentration. Finally, the effects of ambroxol and levofloxacin in different concentrations on the inhibition formation test and removal test of Klebsiella pneumoniae biofilm were observed by crystal violet method combined with confocal laser scanning microscopy. RESULTS: We found that all three groups of bacterial biofilms became mature on the 5th day, and the sensitive group was easier to form and produce biofilm more than the ESBLs and CRKP groups (F=3.725, P=0.032). It was showed that the geometric average of levofloxacin MIC value in the three groups decreased significantly. And the selected strains all showed a synergistic effect on the two-drug combination.In the biofilm test, as the concentration of ambroxol increased, its inhibition rate reached more than 75%, but its biofilm removal rate did not reach 70%. CONCLUSION: Our study support that ambroxol combined with levofloxacin can inhibit the bacterial biofilm of Klebsiella pneumoniae early, and its optimal synergistic concentration is 0.49 mg/mL (ambroxol) + 4 μg/mL (levofloxacin).

Trichosporon asahii ankle cavity effusion infection in a patient with severe aplastic anemia

Rationale: Trichosporon, an anamorphic fungus, proliferates under high humidity, causing serious opportunistic infections collectively called trichosporonosis. Among the Trichosporon species causing trichosporonosis are Trichosporon (T.) asahii, T. asteroides, T. cutaneum etc. Patient concerns: A 38-year-old Chinese male with severe aplastic anemia was admitted due to multiple joints pain, poor appetite, and right ankle swelling. One year earlier he had undergone allogeneic hematopoietic stem cell transplantation. Diagnosis: T. asahii infection and severe aplastic anemia. Interventions: Combined treatment of amphotericin B liposomes (55 mg/24 h) and voriconazole (200 mg/12 h) for 8 days. Outcomes: The symptoms of the patient's ankle were relieved and effusion cultures showed no T. asahii. Lessons: To the best of our knowledge, T. asahii ankle cavity effusion infections are rare. Trichosporon infections may be attributed to risk factors such as improper long-term use of antimicrobials for an underlying disease (e.g., anemia, hypoalbuminemia). Attention should be paid to prevent and control Trichosporon infections in order to avoid comorbidities.

Value of cell-bound complement activation products in the diagnosis of systemic lupus erythematosus / 临床检验杂志

Objective@#To investigate the values of T lymphocyte-bound complement activation products such as T-C3d and T-C4d, B lymphocyte-bound complement activation products such as B-C3d and B-C4d and erythrocyte-bound complement activation products such as E-C3d and E-C4d in the diagnosis of systemic lupus erythematosus (SLE). @*Methods@#Peripheral blood samples from 68 SLE patients, 70 patients with non-SLE autoimmune diseases and 68 healthy controls were collected randomly, and the expression levels of T-C4d, B-C4d, E-C4d, T-C3d, B-C3d and E-C3d in these samples were detected by flow cytometry. Meanwhile, antinuclear antibodies (ANA), anti-double stranded DNA antibodies (anti-dsDNA), peripheral blood cell count and other markers were also detected. The differences of cell-bound complement activation products in three groups were analyzed with the area under the receiver operating characteristic curve (AUC), nonparametric test, sensitivity and specificity. @*Results@#The specific median fluorescence intensity (SMFI) of T-C4d, B-C4d, E-C4d, T-C3d, B-C3d and E-C3d in SLE patients were significantly higher than those in the patients with non-SLE autoimmune diseases and healthy controls (all P＜0.05). The SMFI (median \[P 25, P 75\]) of T-C4d, B-C4d and E-C4d in SLE patients were 3.8(1.2, 13.1), 22.1(6.2, 67.9) and 19.6(1.8, 52.4), respectively. The SMFI of T-C4d, B-C4d and E-C4d in SLE patients with reduced red blood cells and/or lymphocytes were significantly higher than that with normal red blood cell and lymphocyte count (all P＜0.05). The AUCs of T-C4d, B-C4d, E-C4d, T-C3d, B-C3d and E-C3d were 0.711, 0.763, 0.663, 0.631, 0.611 and 0.615, respectively (all P＜0.05). The sensitivity of the combination of T-C4d with B-C4d (73.5%) in the diagnosis of SLE was superior to that of anti-dsDNA (36.8%). @*Conclusion@#The cell-bound complement activation products (CB-CAPs) are specifically expressed in SLE patients, and their combination detection is helpful for the diagnosis of SLE.

Effects of lobaplatin on proliferation and invasion of cervical cancer CaSki cells / 国际肿瘤学杂志

Objective To investigate the effects of lobaplatin on proliferation and invasion of cervical cancer CaSki cells.Methods Human cervical cancer CaSki cells were randomly divided into blank control group,2,6 and 12 μg/ml lobaplatin groups by random number table method.The proliferations of the cells were detected by methyl thiazolyl tetrazolium (MTT).The morphological changes of the cells were observed by inverted microscope.The invasive abilities of the cells were detected by Transwell invasion test.The protein expressions of extracellular signal-regulated kinase (ERK) and phospho-extracellular signal-regulated kinase (p-ERK) were detected by Western blotting.Results The absorbance (A) values of blank group,2,6,12 μg/ml lobaplatin groups cultured for 24 h were 0.513 ± 0.023,0.428 ± 0.014,0.380 ± 0.012 and 0.300 ± 0.013 respectively,those of the cells cultured for 48 h were 0.831 ± 0.024,0.558 ± 0.019,0.415 ± 0.015 and 0.088 ±0.009 respectively,and those of the cells cultured for 72 h were 1.153 ±0.022,0.572 ± 0.023,0.201 ± 0.017 and 0.052 ± 0.014 respectively.The differences were statistically significant (F =12.922,P ＜ 0.001;F =10.192,P ＜ 0.001;F =11.192,P ＜ 0.001),and the differences between each two groups were statistically significant (all P ＜ 0.05).Under inverted microscope,the cells of the platinum groups were shrunken and round,the volume and quantity were reduced,the morphology was irregular,the gap was increased,and the changes were more obvious with the increase of the concentration and the culture time.The numbers of penetrating cells of the blank group,2,6,12 μg/ml lobaplatin groups were 87.27 ±9.38,71.02 ± 8.92,53.20 ± 10.02 and 21.02 ± 7.37 respectively.The difference was statistically significant (F =87.291,P ＜ 0.001),and the differences between each two groups were statistically significant (all P ＜ 0.05).The A values of ERK protein in the blank group,2,6,12 μ~ml lobaplatin groups (0.955 ± 0.021、0.953 ± 0.023、0.950 ± 0.020、0.951 ±0.022)showed no significant difference (F =2.033,P =0.783),but the A values of p-ERK protein in the four groups were 0.941 ±0.015,0.831 ±0.020,0.620 ±0.019 and 0.493 ±0.017 respectively,which showed significant difference (F =11.921,P ＜0.001),and the differences between each two groups were statistically significant (all P ＜ 0.05).Conclusion Lobaplatin can inhibit the proliferation and invasion of cervical cancer CaSki cells,which may be related to the inhibition of the expression of p-ERK protein.

Clinical application of barium radiography and computed tomography-based short-term outcome evaluation criteria in esophageal cancer / 中华放射肿瘤学杂志

Objective To explore and improve the feasibility and prognostic value of barium radiography and computed tomography (CT)-based evaluation criteria in evaluation of the short-term efficacy of radiotherapy for esophageal cancer,and to provide a basis for clinical application.Methods The short-term treatment outcomes of 529 patients with esophageal carcinoma receiving three-dimensional radiotherapy from 2004 to 2015 were evaluated by the 2013 version of barium radiography and CT-based evaluation criteria.The local control (LC) and survival rates were calculated using the Kaplan-Meier method.The log-rank test was used for data analysis and univariate prognostic analysis.The agreement between two evaluation criteria was measured by the Kappa coefficient.Results According to the results of the survival analysis in all the patients using the evaluation criteria for short-term treatment outcomes,the 3-,5-,7-,and 9-year LC rates were 78.6％,69.8％,69.8％,and 63.4％ in the complete response (CR) group (n=52),and 56.4％,47.9％,46.2％,and 42.4％ in the partial response (PR) group (n=409),respectively;the 3-,5-,7-,and 9-year overall survival (OS) rates were 62.7％,49.1％,39.8％,and 39.8％ in the CR group,and 29.5％,21.6％,20.6％,and 19.5％ in the PR group,respectively;the median OS time was 50,17,and 5 months in the CR group,PR group,and non-response group (n=12),respectively (P=0.000).According to CT measurements,the short diameter of residual metastatic lymph node after radiotherapy was between 0.37-3.40 cm (median value=0.82 em).All patients were divided into groups based on the short diameter of residual metastatic lymph node after radiotherapy with a gradient of 0.5 cm.Patients with short diameters of residual metastatic lymph node of ≤ 1.00 cm had a significantly higher OS rate than those with short diameters of residual metastatic lymph node of＞ 1.00 cm (P =0.000).The lymph node volume of 1.00 cm3 in the original criteria was replaced by the short diameter of residual metastatic lymph node of 1.00 cm after radiotherapy and treatment outcomes were re-evaluated using the new criteria.The CR group still had significantly higher LC and OS rates than the PR group (P=0.000).There was a good agreement between the two evaluation criteria (Kappa =0.863).Conclusions The barium radiography and CT-based evaluation criteria for short-term treatment outcomes can accurately evaluate the short-term outcomes and predict prognosis in patients with esophageal carcinoma.Replacing the volume in the original criteria with the short diameter of residual metastatic lymph node after radiotherapy achieves similar results in prognostic prediction.

SCF E3 ubiquitin ligase targets the tumor suppressor ZNRF3 for ubiquitination and degradation

Wnt signaling has emerged as a major regulator of tissue development by governing the self-renewal and maintenance of stem cells in most tissue types. As a key upstream regulator of the Wnt pathway, the transmembrane E3 ligase ZNRF3 has recently been established to play a role in negative regulation of Wnt signaling by targeting Frizzled (FZD) receptor for ubiquitination and degradation. However, the upstream regulation of ZNRF3, in particular the turnover of ZNRF3, is still unclear. Here we report that ZNRF3 is accumulated in the presence of proteasome inhibitor treatment independent of its E3-ubiquitin ligase activity. Furthermore, the Cullin 1-specific SCF complex containing β-TRCP has been identified to directly interact with and ubiquitinate ZNRF3 thereby regulating its protein stability. Similar with the degradation of β-catenin by β-TRCP, ZNRF3 is ubiquitinated by β-TRCP in both CKI-phosphorylation- and degron-dependent manners. Thus, our findings not only identify a novel substrate for β-TRCP oncogenic regulation, but also highlight the dual regulation of Wnt signaling by β-TRCP in a context-dependent manner where β-TRCP negatively regulates Wnt signaling by targeting β-catenin, and positively regulates Wnt signaling by targeting ZNRF3.

Reliability and validity of the Chinese version on Alcohol Use Disorders Identification Test / 中华流行病学杂志

Objective To assess the reliability and validity of the Chinese version on Alcohol Use Disorders Identification Test (AUDIT) among medical students in China and to provide correct way of application on the recommended scales.Methods An E-questionnaire was developed and sent to medical students in five different colleges.Students were all active volunteers to accept the testings.Cronbach's α and split-half reliability were calculated to evaluate the reliability of AUDIT while content,contract,discriminant and convergent validity were performed to measure the validity of the scales.Results The overall Cronbach's α of AUDIT was 0.782 and the split-half reliability was 0.711.Data showed that the domain Cronbach's α and split-half reliability were 0.796 and 0.794 for hazardous alcohol use,0.561 and 0.623 for dependence symptoms,and 0.647 and 0.640 for harmful alcohol use.Results also showed that the content validity index on the levels of items I-CVI)were from 0.83 to 1.00,the content validity index of scale level (S-CVI/UA) was 0.90,content validity index of average scale level (S-CVI/Ave) was 0.99 and the content validity ratios (CVR) were from 0.80 to 1.00.The simplified version of AUDIT supported a presupposed three-factor structure which could explain 61.175％ of the total variance revealed through exploratory factor analysis.AUDIT semed to have good convergent and discriminant validity,with the success rate of calibration experiment as 100％.Conclusion AUDIT showed good reliability and validity among medical students in China thus worth for promotion on its use.

Reliability and validity of the Chinese version on Alcohol Use Disorders Identification Test / 中华流行病学杂志

Objective To assess the reliability and validity of the Chinese version on Alcohol Use Disorders Identification Test (AUDIT) among medical students in China and to provide correct way of application on the recommended scales.Methods An E-questionnaire was developed and sent to medical students in five different colleges.Students were all active volunteers to accept the testings.Cronbach's α and split-half reliability were calculated to evaluate the reliability of AUDIT while content,contract,discriminant and convergent validity were performed to measure the validity of the scales.Results The overall Cronbach's α of AUDIT was 0.782 and the split-half reliability was 0.711.Data showed that the domain Cronbach's α and split-half reliability were 0.796 and 0.794 for hazardous alcohol use,0.561 and 0.623 for dependence symptoms,and 0.647 and 0.640 for harmful alcohol use.Results also showed that the content validity index on the levels of items I-CVI)were from 0.83 to 1.00,the content validity index of scale level (S-CVI/UA) was 0.90,content validity index of average scale level (S-CVI/Ave) was 0.99 and the content validity ratios (CVR) were from 0.80 to 1.00.The simplified version of AUDIT supported a presupposed three-factor structure which could explain 61.175％ of the total variance revealed through exploratory factor analysis.AUDIT semed to have good convergent and discriminant validity,with the success rate of calibration experiment as 100％.Conclusion AUDIT showed good reliability and validity among medical students in China thus worth for promotion on its use.

Incidence and mortality of acute kidney injury in coronary care unit: a retrospective study from a single center / 中华肾脏病杂志

Objective To evaluate the incidence and mortality of acute kidney injury (AKI) in coronary care unit (CCU),and to identify the risk factors of the incidence of AKI and the mortality of CCU patients.Methods A total of 414 patients in CCU from January 1,2014 to June 1,2015 at Zhongnan Hospital of Wuhan University were enrolled.Based on the KDIGO-AKI criteria,these patients were classified into two groups:NAKI group (patients without AKI) and AKI group.Clinical characteristics and laboratory data of two groups were compared.The risk factors of the incidence of AKI and the mortality of CCU patients was analyzed by logistic regression,and then the receiver operating characteristic (ROC) curve was drawn to evaluate the predictive value of these risk factors.Results (1) Among 414 patients,136(32.9％) patients fulfilled the criteria for AKI,and 14.0％ patients in AKI stage 1,10.9％ in AKI stage 2 and 8.0％ in AKI stage 3.(2) The total CCU mortality was 15.0％.Mortality of AKI patients in the CCU was 33.3％,higher than 6.1％ in patients without AKI (OR=7.735,95％CI 4.215-14.196,P ＜ 0.001).The mortality worsened with increasing severity of AKI (22.4％ for AKI stage 1 group,37.8％ for AKI stage 2 group,45.4％ for AKI stage 3 group).(3) Anemia (OR=8.274,95％ CI 4.363-15.689),history of chronic illness (OR=2.582,95％ CI 1.400-4.760),APACHE]Ⅱ seores (OR=1.813,95％CI 1.739-1.895),male (OR=3.666,95％CI 1.860-7.226) were the independent risk factors for AKI,while the normal mean arterial pressure (MAP) (OR=0.292,95％CI 0.153-0.556) and normal estimated glonerular filtration rate (eGFR) (OR=0.166,95％CI 0.090-0.306) are the protective factors for AKI (all P ＜ 0.05).(4) AKI was the most powerful independent factor associated with the mortality of CCU patients (OR=7.050,95％ CI 2.970-16.735,P ＜ 0.001).Other independent risk factors for CCU mortality included history of chronic illness,ejection fraction and APACHE Ⅱ ≥ 15 scores (all P ＜ 0.05),while the normal MAP and normal eGFR were the protective factors (all P ＜ 0.05).(5) For predicting AKI,eGFR displayed an excellent areas under the ROC curve (AUC=0.815,P ＜ 0.001),and for CCU mortality,APACHE Ⅱ scores had the highest overall correctness of prediction (AUC=0.757 P ＜ 0.001).Conclusions CCU patients have high morbidity of AKI,which is the most powerful independent factor associated with the increased CCU mortality.The eGFR is the best predictor for AKI,and then through the evaluation of eGFR for CCU patients,we can evaluate high-risk groups,make early interventions and then improve the prognosis of CCU patients.

Long term survival analysis of middle and lower thoracic esophageal carcinoma of stage T4 N (+) treated with 3DRT / 中华放射肿瘤学杂志

Objective To observe the long?term survival and adverse reactions in patients with stage T4 N (+) Ⅲ middle and lower thoracic esophageal carcinoma undergoing intensity?modulated radiotherapy ( IMRT) . Methods From 2004 to 2010, 300 patients with stage T4 N (+) Ⅲ middle and lower thoracic esophageal carcinoma, consisting of 202 treated with three?dimensional conformal radiotherapy ( 3DCRT ) and 98 treated with IMRT, were enrolled as subjects. All patients received conventionally fractionated radiotherapy with a prescribed dose of 60 Gy. The long?term survival and adverse reactions were compared between patients treated with the two different radiotherapy regimens. The survival rates were calculated by the Kaplan?Meier method and analyzed by the log?rank test. Results The 5?and 7?year sample sizes were 239 and 120, respectively. The 3DCRT group had significantly lower 1?, 3?, 5?, and 7?year local control (LC) and overall survival (OS) rates than the IMRT group (64. 4% vs. 68. 3%, 40. 6% vs. 55. 3%, 38. 3% vs. 51. 9%, 34. 2% vs. 51. 9%, P=0. 048;54. 5% vs. 63. 3%, 19. 8% vs. 34. 7%, 14. 7% vs. 24. 4%, 10. 9% vs. 20. 3%, P=0. 013) . The stratified analysis showed that for patients older than 65 years, with the length of esophageal lesion>8. 0 cm before radiotherapy, the largest diameter of esophageal lesion in computed tomography image>4. 6 cm, gross tumor volume ( GTV)>60 cm3 , metastases to adjacent tissues or organs, stage N2 , and without chemotherapy, the IMRT group had a significantly higher OS rate than the 3DCRT group (P=0. 022,0. 003,0. 022,0. 034,0. 016,0. 044,0. 047). The GTV Dmin and GTVD100 were significantly higher in the IMRT group than in the 3DCRT group ( P=0. 000,0. 000) , while the Dmax of the spinal cord was significantly lower in the IMRT group than in the 3DCRT group ( P=0. 000) . Compared with the 3DCRT group, the IMRT group had a significantly higher incidence of acute radiation?induced esophagitis, particularly grade 1?2 esophagitis (P=0. 000). The mortality rate caused by local tumor was significantly higher in the 3DCRT group than in the IMRT group ( P= 0. 039 ) . Conclusions In the treatment of locally advanced middle and lower thoracic esophageal carcinoma, IMRT is safe and effective;it significantly improves the LC rate and long?term survival without severe toxicity to normal tissues. The results of this retrospective study need to be confirmed by prospective randomized controlled studies.

Efficacy of sustained releasing larvicides package against larval breeding: a field trial / 中华地方病学杂志

Objective To evaluate the efficacy of a sustained releasing mosquito larvicide package against larval breeding and its impact on water and plant,in order to provide a scientific evidence for its application in control and prevention of Dengue.Methods Guangzhou Center for Disease Control and Prevention was chosen as the test place.Twenty test sites were set up,2 bags of sustained releasing larvicides package,1 bag of sustained releasing larvicides package,3 g 1％ temephos granules and nothing were put into 4 glass bottles for each test site from July to December in 2014,respectively.The 4 glass bottles were called high dose (H) group,low dose (L)group,positive control (P) group and blank control (B) group,respectively.The 4 groups were observed at intervals of 10 days for 19 times.Environmental air temperature,turbidity of water,number of larvae and damage of plant were recorded.And 5 test sites were selected to collect water specimen.The chemical oxygen demand,ammonia nitrogen concentration and temephos concentration of water specimen were detected.Results The larval breeding rates were 0 (0/380),1.1％ (4/380),0.8％ (3/380) and 63.4％ (241/380),damage rates of plant were 5.0％ (19/380),5.5％ (21/380),4.7％ (18/380),4.7％ (18/380) and turbidty rates of water were 24.5％ (93/380),19.7％ (75/380),33.4％ (127/380) and 20.3％ (77/380) in H,L,P and B groups,respectively.Statistically significant differences were seen in larval breeding rate and turbidity rate of water between different groups (x2 =823.565,24.715,all P ＜ 0.05),but they were not seen in damage rate of plant (x2 =0.332,P ＞ 0.05).The temephos concentrations were 1.24,0.78 and 2.33 mg/L in H,L and P groups,respectively.Statistically significant differences were seen in temephos concentration between different groups (H =35.426,P ＜ 0.01),but they were not seen in chemical oxygen demand and ammonia nitrogen concentration (H =0.239,0.013,all P ＞ 0.05).Conclusions The sustained releasing package of mosquito larvicide makes less pollution to water and has no impact on water turbidity.Moreover,it doesn't damage the aquatic plant.The efficacy of the sustained releasing package of mosquito larvicide could effectively prevent mosquito larval breeding in Dengue epidemic period.

Optimal radiation dose for esophageal squamous cell carcinoma / 中华放射肿瘤学杂志

Objective To analyze the survival of patients with esophageal squamous cell carcinoma (ESCC)treated by different regimens and different radiation doses and to explore the optimal radiation dose and subgroups with potential clinical benefit. Methods A total of 1387 patients with ESCC who received conformal radiotherapy or intensity-modulated radiotherapy in our hospital from July 2003 to March 2014 were enrolled in this retrospective study. The patients who received different radiation doses in radiotherapy alone or in concurrent chemoradiotherapy were analyzed.The log-rank test and Cox regression analysis were used to explore the optimal radiation dose and the benefited subgroups. Results A total of 780 patients only received radiotherapy. Among them,the median survival of patients receiving radiation dose<60 Gy(n=91),60 Gy(n=429),and>60 Gy(n=260)was 9,20,and 23 months,respectively,suggesting a significant difference(P=0.000).The patients with a radiation dose of 60 Gy had a similar survival curve to the patients with radiation dose>60 Gy,both significantly higher than that in patients with radiation dose<60 Gy (P=0.000,0.000).Totally 302 patients received concurrent chemoradiotherapy. Among them,the median survival of patients receiving radiation dose<60 Gy(n=18),60 Gy(n=224),and>60 Gy(n=60)was 22, 34,and 15 months,respectively,suggesting a significant difference(P=0.004).The survival curve showed no significant difference between the patients with radiation dose<60 Gy and>60 Gy(P=0.952),while the patients with a radiation dose of 60 Gy had a better survival compared with the patients with radiation dose<60 Gy or>60 Gy. The Cox multivariate regression analysis indicated that the ESCC patients receiving radiotherapy alone or concurrent chemoradiotherapy had different prognosis;gross tumor volume(GTV)and radiation dose were two independent prognostic factors in the same treatment model(P=0.045,0.001).In radiotherapy alone,radiation dose ≥60 Gy was a protective factor for the patients' survival(P=0.000).In concurrent chemoradiotherapy,a radiation dose of 60 Gy was a protective factor,while radiation dose<60 Gy or>60 Gy presented no survival benefit(P=0.051). Conclusions The optimal radiation dose is no less than 60 Gy in ESCC patients treated by radiotherapy alone. If the patients receive concurrent chemoradiotherapy,the radiation dose of 60 Gy is recommended.