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【Objective】 To form the sampling data interval by retrospectively analyzing the sampling data of quality monitoring of fresh frozen plasma, cryoprecipitates and leukocyte-free platelets in all blood stations in Hebei Province during the past 7 years. 【Methods】 The data of blood component sampling from 12 blood station quality control laboratories in Hebei from 2015 to 2021 were collected. The FⅧ content and plasma protein content of fresh frozen plasma, the FⅧ content and fibrinogen content of cryoprecipitates, and the leukocyte residuals, red blood cell mixed and platelet content of leukocyte-free platelets were taken as the objects for discrete point and fitted curve analysis. 【Results】 The FⅧ level of fresh frozen plasma: (1.36±1.1) IU/mL, 5 blood stations showed a representative overall high or low or fluctuated characteristics; Fresh frozen plasm-plasma protein items: overall mean ±SD: (61.13±16.7) g/L, four blood stations showed scattered distribution or continuous high value scattered points; Cryoprecipitates FⅧ: the overall mean ±SD: (134.25±58.7) IU/mL, four blood stations showed the differentiation characteristics of continuous high, low or stable in the middle; Cryoprecipitates-fibrinogen items: the overall mean ±SD: (215.27±83.5) mg, five blood stations showed the overall high or low and fluctuated. Leukocyte-free apheresis platelet-to-leukocyte residual items: overall mean ±SD: 0.37±0.96 (×106/bag), two blood stations showed a relatively high representative overall characteristics, and the rest were concentrated between 0 and 1; The total mean ±SD of platelet-to-red blood cell mixture without leukocyte was 2.45±2.82 (×109/bag), with obvious segmented concentrated distribution, and scattered distribution in 3 blood centers. Platelet content: the overall mean ±SD was 3.14±1.55 (×1011/bag), many deviations were noticed in 3 blood stations, and 1 blood station showed representative overall high characteristics. 【Conclusion】 This analysis shows that the distribution status of each blood station in different items is similar. The distribution status of discrete point groups and the change trend of the concentrated part of the fitting curve show that there are some differences in the monitoring level between the quality control laboratories of each blood station, and the update of detection instruments and reagents and the selection of detection methods greatly affect the test results. The summary data presented the index interval framework formed in the past 7 years, which helped to understand the difference between the results of each laboratory, correct the accuracy of the test results, better play the guiding role of quality monitoring in the blood preparation process, and continue to enhance the standardization of the whole process of blood collection and supply in the province.
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【Objective】 To conduct the laboratory quality assessment between 12 blood stations in Hebei province, analyze the results and explore the accuracy and comparability of testing, so as to improve the level of testing ability and quality management. 【Methods】 With reference to the external quality assessment rules of National Center for Clinical Laboratories and combined with the instructions of quality assessment samples, daily testing process of the laboratories were assessed. The quality indicators include blood cell count (WBC, RBC, Hb, HCT, MCV, MCH, MCHC and PLT), biochemical items (TP) and coagulation parameters (FIB and FⅧ). 【Results】 There are still problems in laboratories in terms of personnel operation, instrument maintenance and the impact of different reagent batches, especially in biochemical items and coagulation parameters. The pass rate of biochemical items was the lowest, only 72.75%, and that of blood cell count was the highest, reaching 98.75%. 【Conclusion】 With the progress of the project, the quality monitoring level of daily blood sampling tests in the quality control laboratory of each blood station has been improved. However, it is still necessary for each laboratory to improve the testing ability and quality management to a higher level in Hebei.
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Objective:To investigate the clinical application of free/total prostate-specific antigen (f/tPSA), peripheral blood neutrophil-to-lymphocyte ratio (NLR), interleukin-6 (IL-6) and prostate health index density (PHID) detection in the early diagnosis of prostate cancer.Methods:The clinical data of 160 patients with abnormal prostate specific antigen (PSA) who were admitted to the Second Affiliated Hospital of Xuzhou Medical University from January 2020 to January 2022 were retrospectively analyzed. According to the pathological results of prostate biopsy or electrical resection, the patients were divided into prostate cancer group (68 cases) and benign prostatic hyperplasia group (92 cases), and 50 male healthy physical examiners in the Second Affiliated Hospital of Xuzhou Medical University during the same period were selected as healthy control group. All enrolled members were tested for total prostate-specific antigen (tPSA), free prostate-specific antigen (fPSA), and prostate specific antigen isoform 2 (p2PSA), IL-6 and other indicators, and the f/tPSA, prostate health index (PHI), PHID and NLR were calculated. Receiver operating characteristic (ROC) curve was plotted to compare the efficacy of each index in diagnosing and differentially diagnosing prostate cancer and benign prostatic hyperplasia.Results:The serum levels of tPSA, fPSA, p2PSA, PHI and PHID in the prostate cancer group were higher than those in the benign prostatic hyperplasia group and the healthy control group (all P < 0.05), and the serum f/tPSA was lower than that in the benign prostatic hyperplasia group and the healthy control group ( P < 0.05). The area under the curve (AUC) of PHID for the diagnosis of early stage prostate cancer was the largest [0.915 (95% CI 0.864-0.966)], followed by PHI [0.884 (95% CI 0.823-0.944)]. The sensitivity of both f/tPSA and PHI in diagnosing early stage prostate cancer was 86.80%, which was higher than other indicators; the specificity of PHID in diagnosing early stage prostate cancer was 94.00%, which was higher than other indicators. The AUC of f/tPSA for the diagnosis of benign prostatic hyperplasia was the largest [0.828 (95% CI 0.739-0.917)], followed by PHID [0.826 (95% CI 0.760-0.892)]. The sensitivity of f/tPSA in diagnosing benign prostatic hyperplasia (85.90%) was higher than other indicators, and the specificity of PHI in diagnosing benign prostatic hyperplasia (94.00%) was higher than other indicators. The AUC of fPSA, PHID, f/tPSA and p2PSA in differentiating early stage prostate cancer and benign prostatic hyperplasia were 0.752 (95% CI 0.663-0.841), 0.730 (95% CI 0.647-0.812), 0.713 (95% CI 0.623-0.803), 0.710 (95% CI 0.629-0.791), respectively, and there was no significant difference in each pairwise comparison (all P > 0.05). The sensitivity of NLR in differentiating early stage prostate cancer and benign prostatic hyperplasia was 91.20%, which was higher than other indicators, and the specificity of fPSA in differentiating early stage prostate cancer and benign prostatic hyperplasia was 94.00%, which was higher than other indicators. Conclusions:The f/tPSA, PHI and PHID detection have certain clinical values in the early diagnosis of prostate cancer, and can provide references for early diagnosis, early treatment and prognosis evaluation of high-risk population of prostate cancer.
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Objective:To construct a quality evaluation index system of pulmonary rehabilitation for lung cancer patients in order to supply reference for evaluation of rehabilitation quality for lung cancer patients.Methods:This research adopted Delphi technique of non-experimental study. A total of 25 experts were invited to participate 2 rounds of letter enquiry from January to April in 2022. Literature retrieval, group discussion and Delphi method were used to establish the evaluation system of pulmonary rehabilitation for patients with lung cancer based on the social ecology theory.Results:The recovery rates of the two rounds of expert correspondence were 84%(21/25) and 100%(21/21), respectively, the expert authority coefficient was 0.824, and the Kendall′s W value was 0.279 and 0.186, respectively, which were statistically significant ( χ2=409.57, 218.23, both P<0.05). The finally formed quality evaluation index system of pulmonary rehabilitation for lung cancer patients based on social ecology theory included 4 first-level indicators, 11 second-level indicators, and 42 third-level indicators. Conclusions:The evaluation system of pulmonary rehabilitation for lung cancer surgery patients constructed in this study has certain scientificity and practicability, and provides a reference for the continuous improvement of the quality of care for lung cancer patients in our country.
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【Objective】 To investigate the status quo and existing problems of quality control laboratory of blood stations in Hebei province, and to provide reference for the capacity building of quality control laboratory in the future. 【Methods】 The data of routine blood collection and supply, blood component preparation, blood sampling ratio/frequency, and sampling results among 12 blood stations in Hebei in 2020 were collected. The monitoring effect of blood component quality in Hebei province, combined with the indicator changes of main blood components, were analyzed. 【Results】 Blood component preparation from blood stations in Hebei are different. The requirements for the number and frequency of routine sampling in each apartment are not standardized, and retrospective analysis was not conducted basically. Although the frequency and sampling of blood component quality monitoring can meet the minimum requirements of the Quality Monitoring Guidelines of Whole Blood and Blood Components 2017 and Technical Operating Regulations of Blood Stations (2019 Edition), but does not match the actual collection and preparation units. The qualification rate and reliability are unstable due to the small sample size. Monthly quality inspection can result in judgment errors, which is not conducive to systematic analysis and continuous improvement of blood collection and supply process. 【Conclusion】 In the context of continuous enhancement of blood collection and supply standardization in Hebei Province, the blood quality monitoring mechanism based on intra-province consistency can be further studied to standardize intra-province homogenization of blood quality monitoring in multiple links including sampling rules, statistical analysis and data comparison calibers.
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Objective:Summarize and analyze the clinical features of immune checkpoint inhibitor (ICI)-related colitis.Methods:From January 2019 to September 2020, the clinical data of 8 patients with ICI-related colitis from Peking Union Medical College Hospital were retrospectively collected and including the onset of ICI-related colitis, clinical symptoms, endoscopic and pathological findings, treatment, comorbidities and resuming of ICI. Independent sample t test was used for statistical analysis. Results:Eight patients were all male, and the median age (range) was 66 years old (55 to 74 years old), 7 cases were diagnosed with stage Ⅳ lung cancer and 1 case was diagnosed with stage Ⅲc pyelo-carcinoma. Among 8 patients, 4 cases of ICI-related colitis occurred during combination of anti-programmed death-1 (PD-1) treatment and chemotherapy, 2 cases occurred during anti-PD-1 monotherapy after combination of anti-PD-1 treatment and chemotherapy, and 2 cases occurred after anti-PD-1 monotherapy. The median time (range) was 44 d (27 to 128 d) from initial anti-PD-1 treatment to the onset of ICI-related colitis and the median time (range) was 8 d (6 to 35 d) from last anti-PD-1 treatment to onset of ICI-related colitis. The ICI efficacy of 4 patients had partial response, 2 patients had stable disease, 1 patient had disease progression, and 1 patient′s condition was not assessed. All 8 patients had moderate to severe extensive colitis. The main clinical manifestation was diarrhea (5/8), 3 patients accompanied by abdominal pain. The endoscopic findings were diffuse mucosal erosion, accompanied by ulcer in 2 patients. The main pathologic findings were cryptitis or crypt abscess, accompanied by apoptosis in 2 patients. Eight patients were all treated with glucocorticoids, among them 2 patients were further treated with biologics, due to the insufficient efficacy of glucocorticoid treatment, 4 patients had opportunistic infections. The initial prednisone dose for patients with opportunistic infections and patients without opportunistic infections was (85.00±52.60) and (60.00±23.09) mg, respectively. The prednisone treatment course was (8.75±4.03) and (7.50±3.11) weeks, respectively, and the differences were not statistically significant (both P>0.05). The colitis relapsed in all 3 patients after resuming of ICI. Conclusions:ICI-related colitis had corresponding ICI treatment history and clinical, endoscopic, and histopathological features. Glucocorticoid is the main treatment, and it is prone to relapse after resuming of ICI.
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Objective:To investigate the clinical significance of changes in heparin binding protein(HBP), procalcitonin (PCT) and prealbumin (PA) levels in the early diagnosis of intracranial infection in patients with brain tumors after surgery.Methods:The clinical data of 160 patients with brain tumors who underwent surgical treatment in the Second Affiliated Hospital of Xuzhou Medical University from January 2015 to December 2020 were retrospectively analyzed. And 80 cases of intracranial infection after surgery were classified as the infection group. According to the severity of infection, patients were divided into 33 cases in the mild infection group, 36 cases in the moderate infection group, and 11 cases in the severe infection group; 80 cases without postoperative intracranial infection were classified as the non-infection group. All enrolled members were tested for HBP, PCT, PA at time points of 0 h before surgery, 12 h after surgery, 3 d after surgery, and 6 d after surgery. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficacy of intracranial infection in patients with brain tumors after surgery.Results:The levels of HBP and PCT in the infected group were higher than those in the non-infected group before and after surgery at all time points (all P < 0.05). The level of PA in the infected group was lower than that in the non-infected group (all P < 0.05). The levels of HBP and PCT in both groups at various time points after surgery were higher than those before surgery (all P < 0.05), which showed a decreasing trend over time. The level of PA of both groups at all time points after surgery was lower than that before surgery (all P < 0.05), which showed a trend of first decreasing and increasing and then decreasing over time. The preoperative HBP level in the severe infection group [35.45 ng/ml (23.61 ng/ml, 59.44 ng/ml)] was higher than that in the mild infection group [12.51 ng/ml (5.11 ng/ml, 19.92 ng/ml)] and the moderate infection group [22.22 ng/ml (16.49 ng/ml, 27.55 ng/ml)], and differences were statistically significant ( Z = 41.167, 19.105, both P < 0.05). The preoperative PCT level in the severe infection group [1.50 μg/L (1.27 μg/L, 2.44 μg/L)] was higher than that in the mild infection group [0.53 μg/L (0.25 μg/L, 0.85 μg/L)] and the moderate infection group [0.90 μg/L (0.47 μg/L, 1.42 μg/L)], and differences were statistically significant ( Z = 36.167, 14.875; both P < 0.05). The preoperative PA level in the severe infection group [159.22 mg/L (141.61 mg/L, 191.79 mg/L)] was lower than that in the mild infection group [215.91 mg/L (195.21 mg/L, 239.90 mg/L)] and the moderate infection group [194.48 mg/L (178.40 mg/L, 207.60 mg/L)] ( Z = 35.955, 19.567, both P < 0.05). The levels of HBP and PCT before operation were positively correlated with the degree of infection ( r = 0.637, 0.485, both P < 0.01). The preoperative level of PA was negatively correlated with the degree of infection ( r = -0.576, P < 0.01). The preoperative single index detection showed that the maximum the area of the curve (AUC) of postoperative intracranial infection in patients with brain tumors predicted by PA was 0.808 (95% CI 0.741-0.874). The highest specificity of intracranial infection in patients with brain tumors predicted by HBP was 96.3%. The AUC of postoperative intracranial infection in patients with brain tumors predicted by the combination of the three tests was 0.892 (95% CI 0.839-0.944), which was greater than that predicted by other single indicators, and the sensitivity was the highest (86.3%). The AUC of postoperative intracranial infection in patients with brain tumors diagnosed by PCT at 12 h after surgery was maximum [0.804 (95% CI 0.734-0.874)] when predicted by other single indicators. The highest specificity of postoperative intracranial infection in patients with brain tumors diagnosed by HBP was 98.6%. The AUC of postoperative intracranial infection in patients with brain tumors diagnosed by the combination of the three tests was 0.895 (95% CI 0.840-0.950), which was greater than that diagnosed by other single indicators, and the highest sensitivity was 85.0%. Conclusion:The levels of HBP, PCT and PA can provide a reference for the early diagnosis of postoperative intracranial aseptic inflammation in patients with brain tumors. The combined diagnosis of HBP, PCT and PA can better diagnose the postoperative intracranial infection in patients with brain tumors.
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Objective:To explore the significance of combined detection of biomarkers in the diagnosis of early stage primary liver cancer.Methods:A retrospective study model was adopted to analyze 60 patients who were hospitalized from January 2018 to September 2019 in General Hospital of Xuzhou Mining Group and were pathologically diagnosed as early stage primary liver cancer (early stage liver cancer group), and 60 patients with liver cirrhosis (liver cirrhosis group), 30 patients with hepatitis (hepatitis group), 30 healthy examiners (healthy control group) who were admitted during the same period were selected. All participants were tested for liver cancer biomarkers such as alpha-fetoprotein (AFP), vitamin K deficiency Ⅱ-inducing protein (PIVKA-Ⅱ), alpha-fetoprotein variant (AFP-L3) and Golgi protein 73 (GP73). The receiver operating characteristic curve (ROC) was used to compare the differences in the diagnosis of early stage primary liver cancer between individual and combined detection of different indicators.Results:The levels of AFP-L3, AFP, PIVKA-Ⅱ and GP73 in the early stage liver cancer group were higher than those in the cirrhosis group, hepatitis group and healthy control group, and the differences were statistically significant (all P < 0.05). PIVKA-Ⅱ had the largest area under the receiver operator characteristic curve (AUC) for diagnosing early stage liver cancer, which was 0.776; AFP-L3 had the highest specificity for diagnosing early stage liver cancer, which was 82.2%; GP73 had the highest sensitivity for diagnosing early stage liver cancer, which was 80.0%. The AUC of the four indicators in the diagnosis of early stage liver cancer was 0.922, which was higher than a single indicator, and the sensitivity reached 91.7%. Conclusions:Liver cancer biomarkers AFP-L3, PIVKA-Ⅱ, AFP and GP73 all have certain diagnostic value in the diagnosis of early stage primary liver cancer. The combined detection has important clinical significances for diagnosis of early stage primary liver cancer and assessment of liver damage.
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Objective:To analyze clinical characteristics and monitor microbiome changes in patients with anti-PD-1 associated colitis.Methods:Two patients with non-small cell lung cancer who developed colitis after treated with anti-PD-1 antibodies were retrospectively analyzed in Peking Union Medical College Hospital from January 2019 to January 2020. The clinical symptoms, endoscopic and pathological manifestations, as well microbiome changes were analyzed and compared during pre-treatment, post-treatment and relapse.Results:The main clinical manifestations included diarrhea, elevated inflammatory indicators, colonic mucosal diffuse hyperemic edema with erosion by endoscopy. Changes in the structure of crypts were common pathological characteristics. Glucocorticoids were effective agents, which achieved clinical remission and mucosal healing. The microbiome composition of OTUs was different. After glucocorticoid treatment, the alpha diversity Observed species, Shannon, Simpson, Chao1, ACE indexes all decreased. The Firmicutesdecreased with Bacteroidetesincreasing in phylum level; while the Bacteroides increased with Ruminococcaceaedecreasing in genus level. Lactobacilluswas the potentially beneficial genus. Conclusion:Patients developing anti-PD-1 associated colitis have characteristic clinical and pathological manifestations. Glucocorticoids are effective treatment. The fecal microbiome diversity, relative abundance of major phylum and genus have changed after treatment.
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Objective@#To explore the value of heparin-binding protein (HBP) in early diagnosis of severe infection in silicosis patients.@*Methods@#From January 2017 to June 2018, fifty silicosis patients with severe infection and fity without infection were recruited in the Second Affiliated Hospital of Xuzhou Medical University. In the severe infection group, the time of patients diagnosed with severe infection was set as the reference point for time. Blood samples were selected from the hospital inspection system sample library at the time of 24, 48, and 72 hours prior to the reference point. In the non-infection group, blood samples were selected within 24 hours of admission. The blood samples were tested for the levels of HBP, C-reactive protein (CRP), procalcitonin (PCT), and white blood cell (WBC) count. The area under the curve (AUC) of the receiver operating characteristic (ROC) curve, sensitivity, specificity and Youden index (YI) were used to evaluate the diagnostic efficacy of each indicator.@*Results@#The HBP levels at 72, 48, 24, and 0 hours before the diagnosis in the severe infection group were significantly higher than those in the non-infection group (all of the P values <0.001), and decreased with the prolonged time before diagnosis. The ROC curve showed that the AUC of HBP at 72, 48, 24, and 0 h before the diagnosis in the severe infection group [0.828 (0.750-0.907), 0.966 (0.920-0.998), 0.967 (0.961-0.998), 0.997 (0.994-0.999)] was higher than that of PCT [0.563 (0.450-0.677), 0.687 (0.581-0.794), 0.726 (0.622-0.829), 0.982 (0.973-0.986)] and CRP [0.564 (0.449-0.680), 0.648 (0.535-0.761), 0.705 (0.594-0.817), 0.963 (0.924-0.983)] and WBC [0.492 (0.377-0.607), 0.497 (0.383-0.612), 0.628 (0.519-0.738), 0.700 (0.598-0.802)] at the corresponding time. The sensitivity, specificity and YI of HBP were 88.9%-97.6%, 77.1%-98.4% and 0.66-0.96 at 72, 36, 24 and 0 h before diagnosis, respectively.@*Conclusion@#Heparin-binding protein can be used for early diagnosis of severe infection in silicosis patients.
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Immune checkpoint inhibitors (ICIs) have been used more and more Increasingly in clinical oncology treatment, which has significantly improved the prognosis of cancer patients. Over-activation of T cells and related signaling pathways may cause immune-related adverse effects. Renal immune side-effects are relatively rare, but some of them are serious and fatal. This review analyses of the Incidence, clinical and pathological manifestations of ICIs-induced renal injury, and focuses on the differential diagnosis and treatment. Because there are many secondary factors that need to be differentiated from immune mechanism, renal biopsy should be performed if necessary to determine the important decision.
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Immunological checkpoint inhibitors have been approved for a short period of time in China, and real-world clinical data are still in the collection stage. Reports of domestic programmed death-1 (PD-1) treatment-related adverse reactions are rare. The author reported a case of hypotension in the process of Pembrolizumab infusion and successful infusion after blood pressure recovery, hoping to provide reference for the application of immunological checkpoint inhibitors, to provide patients with the greatest clinical benefit.
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Objective@#To investigate the application value of sputum cell DNA ploidy quantitative analysis technique in early non-invasive screening of lung cancer.@*Methods@#The clinical data of 84 patients with lung cancer (lung cancer group) and 84 patients with benign lung disease (lung benign disease group) who were admitted to the Second Affiliated Hospital of Xuzhou Medical University from April 2016 to May 2017 were retrospectively analyzed, and 80 healthy subjects were selected as the control group. The sputum samples of all subjects were collected, and 55 corresponding lavage fluid samples in the lung cancer group were also collected. A fully automated cell tumor screening analysis system was used to make DNA ploidy quantitative analysis in all specimens, and the results were compared with sputum smear and liquid-based thin-layer cytology results.@*Results@#The positive detection rates of routine smear, liquid-based thin-layer cytology and DNA ploidy quantitative analysis in lung cancer group were 4.76% (4/84), 29.76% (25/84) and 45.24% (38/84). The positive detection rate of liquid-based thin-layer cytology and DNA ploidy quantitative analysis was higher than that of routine smear, and the differences were statistically significant (χ 2 = 18.38, P < 0.01; χ2 = 36.70, P < 0.01); the positive detection rate of DNA ploidy quantitative analysis was higher than that of liquid-based thin-layer cytology, and the difference was statistically significant (χ 2 = 4.29, P = 0.038). The positive detection rate of DNA ploidy quantitative analysis in lavage fluid samples was higher than that in sputum samples [65.45% (36/55) vs. 50.91% (28/55)], but the difference was not statistically significant (χ 2 = 2.39, P = 0.122). For peripheral lung cancer patients, the positive detection rate of DNA ploidy quantitative analysis was higher than that of liquid-based thin-layer cytology (χ 2 = 4.55, P = 0.033). The positive detection rate of squamous cell carcinoma [52.17% (24/46)] by using DNA ploidy quantitative analysis was higher than that of adenocarcinoma [36.36% (8/22)], small cell carcinoma [26.36% (4/11)] and adenosquamous carcinoma [40.00% (2/5)], but the differences were not statistically significant (all P > 0.05).@*Conclusions@#DNA ploidy quantitative analysis technique of sputum cells can directly reflect the malignant transformation of early lung cancer patients compared with traditional smear method and liquid-based thin-layer technology. Its positive detection rate is higher than that of the conventional detection method, which can be used as a valuable reference index for early screening of lung cancer.
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Objective To construct and authenticate the lentiviral-mediated overexpression of mouse mitochondrial-targeted-8-oxoguanine DNA-glycosylase 1 (mito-OGG1) gene and the lentiviral-mediated short hairpin RNA (shRNA) down-regulation of OGG1 gene expression model in 661W cells.Methods Constructed the target plasmids,including pLenti-EF1a-EGFP-P2A-Puro-CMV-Mito-OGG1-3Flag (pLenti-OGG1-GFP) and pLKD-CMV-G&PR-U6-shRNA (pLKD-shRNA).293T cells were used to obtain green fluorescent protein (GFP)-tagged lentiviral vector of interest by using a second generation lentivirus packaging system.293T cells were also used for the virus titer estimation.The multiplicity of infection (MOI) of 661W cells was detected by fluorescence microscopy.A stable transfected cell line was screened by puromycin.Immunofluorescence was used to detect transfection efficiency and cytochrome C oxidase Ⅳ (COXⅣ)-OGG1 co-localization.OGG1 mRNA and protein expression levels were detected by real-time qantitative PCR (QPCR) and Western blot.Results Sequencing results showed that the inserted sequence in the over-expression plasmid was consistent with the mouse OGG1 (NM_010957.4) gene sequence in the gene library.The original lentiviral titer after packaging and purification was between 2.0× 107to 6.0× 107 TU/ml.The optimal MOI of 661W cells was 40,and puromycin with a concentration of 4.0 μg/ml successfully screened stable transformation.The transfection efficiency was up to 100％ after screening.Immunofluorescence demonstrated successful co-localization of OGG1 and COXⅣ.The relative expression levels of OGG1 mRNA in the blank control group,OGG1 group,overexpression control group,shRNA group and low expression control group were 1.000±0.000,41.581±12.206,0.888±0.056,0.239±0.121 and 1.081±0.083,and the relative expression levels of OGG1 protein were 1.029±0.153,1.657 ± 0.237,0.752 ± 0.143,0.471 ± 0.149 and 1.036 ± 0.185,respectively,with significant differences between them (F=44.654,30.948;both at P＜0.05),the relative expression levels of OGG1 mRNA and protein in the OGG1 group were significantly higher than those in the overexpression control group,the relative expression levels of OGG1 mRNA and protein in the shRNA group were significantly lower than those in the lower expression control group,with significant differences between them (all at P＜0.05).Conclusions The mitoOGG1 overexpression and OGG1 knockdown models of 661W cells are successfully constructed,which provides the preliminary experimental basis for follow-up study.
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Objective To investigate the application value of sputum cell DNA ploidy quantitative analysis technique in early non-invasive screening of lung cancer. Methods The clinical data of 84 patients with lung cancer (lung cancer group) and 84 patients with benign lung disease (lung benign disease group) who were admitted to the Second Affiliated Hospital of Xuzhou Medical University from April 2016 to May 2017 were retrospectively analyzed, and 80 healthy subjects were selected as the control group. The sputum samples of all subjects were collected, and 55 corresponding lavage fluid samples in the lung cancer group were also collected. A fully automated cell tumor screening analysis system was used to make DNA ploidy quantitative analysis in all specimens, and the results were compared with sputum smear and liquid-based thin-layer cytology results. Results The positive detection rates of routine smear, liquid-based thin-layer cytology and DNA ploidy quantitative analysis in lung cancer group were 4.76％ (4/84), 29.76％ (25/84) and 45.24％ (38/84). The positive detection rate of liquid-based thin-layer cytology and DNA ploidy quantitative analysis was higher than that of routine smear, and the differences were statistically significant (χ2= 18.38, P<0.01;χ2=36.70, P<0.01);the positive detection rate of DNA ploidy quantitative analysis was higher than that of liquid-based thin-layer cytology, and the difference was statistically significant (χ2= 4.29, P= 0.038). The positive detection rate of DNA ploidy quantitative analysis in lavage fluid samples was higher than that in sputum samples [65.45％(36/55) vs. 50.91％ (28/55)], but the difference was not statistically significant (χ2=2.39, P= 0.122). For peripheral lung cancer patients, the positive detection rate of DNA ploidy quantitative analysis was higher than that of liquid-based thin-layer cytology (χ2=4.55, P=0.033). The positive detection rate of squamous cell carcinoma [52.17％ (24/46)] by using DNA ploidy quantitative analysis was higher than that of adenocarcinoma [36.36％ (8/22)], small cell carcinoma [26.36％ (4/11)] and adenosquamous carcinoma [40.00％ (2/5)], but the differences were not statistically significant (all P> 0.05). Conclusions DNA ploidy quantitative analysis technique of sputum cells can directly reflect the malignant transformation of early lung cancer patients compared with traditional smear method and liquid-based thin-layer technology. Its positive detection rate is higher than that of the conventional detection method, which can be used as a valuable reference index for early screening of lung cancer.
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Immune checkpoint inhibitors (ICIs) targeting CTLA-4 and PD-1 /PD-L1 are increasingly used to treat several types of cancer.Although several agents have been approved,only 10％ to 30％ of patients have benefited from them.Some studies have shown that intestinal micro-ecology can affect the outcome of ICIs through immune regulation in cancer patients.This literature review s summarized the regulation and predictive functions of intestinal micro-ecology on ICIs treatment,and introduced possible mechanisms of intestinal micro-ecology that affacted the efficacy of immunotherapy.In addition,some approaches for detecting gut microbiome were also summarized.Gut microbiota highlighted in this review may serve as novel biomarkers to predict clinical outcomes in cancer patients.
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Objective@#To introduce a self-developed bone dust collector designed by the authors and evaluate its efficiency in mastoid obliteration following mastoidectomy.@*Methods@#Consecutive patients, from April 2017 to March 2018, who prepared to receive mastoidectomy were randomly divided into two groups, and in each group the bone dust was harvested by self-developed bone dust collector or by conventional used method respectively in mastoidectomy. The amount of the harvested bone dust and the time consumed in the collecting procedure were compared between two groups. The infection of the bone dust after mastoid obliteration was also evaluated during follow up.@*Results@#33 patients were recruited in bone dust collector group, and 31 patients in conventional method group.There is no significance of difference between two groups in sex ratio, age and pneumatization of mastoid cells (P>0.05 for all). The median amount of bone dust harvested by bone dust collector was significantly larger than that collected by conventional method (1.8 g vs 1.1 g, P<0.05). The median time spent in bone dust collector group was significantly shorter than that spent in conventional method group (4 minutes vs 6 minutes, P<0.05). No bone dust infection was found in the follow-up in all patients.@*Conclusion@#The present self-developed bone dust collector is a easy and useful apparatus which can significantly improve the efficiency of collecting bone dust in mastoidectomy.
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Objective To explore the significance of long non-coding RNA (lncRNA) taurine up-regulated gene 1 (TUG1) in hepatocellular carcinoma (HCC),to predict the target gene of TUG1,and to provide a ref-erence for further study of TUG1 in HCC.Methods The differential expression of TUG1 in HCC was ana-lyzed by using the UALCAN database and the survival analysis of TUG1 was performed.The target gene of TUG1 was predicted by RegRNA 2.0 biology software,HMDD,targetscan and microT-CDS,and the regulato-ry network of lncRNA TUG1-microRNAs-mRNAs was constructed.The predicted target gene was analyzed by Gene Ontology (GO) and KEGG signal transduction pathway enrichment by using FunRich platform. Results TUG1 expression in HCC was significantly increased,and the expression level of TUG1 increased generally with the increase of tumor grade.The overall survival of patients with low expression of lncRNA TUG1 was significantly longer than that of lncRNA TUG1 high expression patients.There were four possible binding sites of HCC related microRNAs (hsa-mir-122-5p,hsa-mir-200a-3p,hsa-mir-34c-3p,hsa-mir-629-3p) on TUG1,which regulated 245 downstream target genes and formed the regulatory network of lncRNA TUG1-microRNAs-mRNAs.In the biological process,microRNA target genes were highly enriched in the processes such as the regulation of nucleobase,nucleoside,nucleotide and nucleic acid metabolism.In KEGG pathway analysis,microRNA target genes were highly enriched to the signal pathways mediated by Syndecan and TRAIL.Conclusion TUG1 expression level in HCC increased.Increased expression of TUG1 is associat-ed with poor prognosis in HCC.Bioinformatics methods can be used to explore the mechanism of tumorigene-sis from the molecular level,which can provide valuable information for subsequent experiments and clinical diagnosis and treatment.
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Objective To evaluate the efficacy and safety of anlotinib in patients with advanced non-small cell lung cancer (NSCLC).Methods Patients with stageⅢB/ⅣNSCLC who progressed after two lines or more regimens were randomized into anlotinib group (12 mg daily from day 1 to 14 of a 21-day cycle) or placebo group with ratio of 2:1. Study drugs or placebo were given until disease progression or intolerable toxicity. The primary endpoint was overall survival (OS), and the second endpoints were progression free survival (PFS), objective response rate, and disease control rate. Results Between April 2015 and December 2015, twenty-four patients were assigned at Peking Union Medical College Hospital. The baseline characteristics of the anlotinib group (n=16) and placebo group (n=8) were fairly comparable. The median OS was 12.7 months in anlotinib group and 11.1 months in placebo group (P=0.460). The median PFS was 4.0 months in anlotinib group and 1.4 months in placebo group (P=0.065). The common adverse events were manageable such as hypertension, hand-foot syndrome, thyroiddy sfunction. No drug-related mortality occurred. Conclusions Anlotinib had a trend of improvement in OS and PFS as third-line treatment or beyond in advanced NSCLC compared with placebo with manageable toxicity.
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Objective To evaluate the effect of methylene blue (MB) on hydrogen peroxide (H2O2)-induced apoptosis in macrophages through mitochondria-dependent pathway in mice.Methods Mouse peritoneal macrophage line RAW264.7 cells were cultured in DMEM culture medium containing 10％ fetal bovine serum.Cells were divided into 6 groups (n =24 each) using a random number table method:control group (group C),H2O2 group,prophylactic different concentrations of MB groups (MB1,2 groups) and therapeutic different concentrations of MB groups (MB3.4 groups).H2O2 50 μmol/L was added to the culture medium in group H2O2.MB was added to the culture medium with the final concentrations of 0.1 μmol/L (in MB1 and MB3 groups) and 1.0 μmol/L (in MB2 and MB4 groups) at 30 min before adding H2O2 in MB1.2 groups and 30 min after adding H2O2 in MB3.4 groups.At 24 h of culture or incubation in each group,the cell survival rate was measured by methyl thiazolyl tetrazolium assay,the activity of reactive oxygen species (ROS) in cells was determined with the fluorescent probe,the lactate dehydrogenase (LDH) activity in supernatant was detected by spectrophotometry,the activity of superoxide dismutase (SOD) in cells was detected by colorimetric method,mitochondrial membrane potential (MMP) was measured using rhodamine 123 staining,the content of ATP was determined by an ATP bioluminescent method,the expression of pro-caspase-3 and spliceosomes P20 protein and P 18 protein was detected by Western blot,and cell apoptosis was detected using flow cytometry.Results Compared with group C,the cell survival rate,SOD activity and contents of MMP and ATP were significantly decreased,the ROS activity and activity of LDH in supernatant were increased,the expression of pro-caspase-3 and spliceosomes P20 protein and P18 protein was up-regulated,and early and late apoptosis rates were increased in the other five groups (P＜0.05).Compared with group H2O2,the cell survival rate,SOD activity and contents of MMP and ATP were significantly increased,the ROS activity and activity of LDH in supernatant were decreased,the expression of pro-caspase-3 and spliceosomes P20 protein and P18 protein was down-regulated,and early and late apoptosis rates were decreased in MB1-4 groups (P＜0.05).Compared with group MB1,the cell survival rate was significantly decreased,and the expression of caspase-3 spliceosome P 18 was down-regulated in group MB2,and the cell survival rate and SOD activity were significantly decreased,and the activity of ROS was increased in group MB3 (P＜0.05).Compared with group MB4,the expression of caspase-3 spliceosome P 18 was significantly down-regulated,early and late apoptosis rates were decreased,and the activity of ROS was increased in group MB2,and the activity of ROS was significantly increased in group MB3 (P＜0.05).Conclusion The mechanism by which MB attenuates H2O2-induced oxidative damage to macrophages is related to inhibiting cell apoptosis in macrophages through mitochondria-dependent pathway in mice.