Objective:To investigate the application value of three-dimensional (3D) visualization combined with indocyanine green (ICG) fluorescence imaging in anatomical hepatectomy for hepatocellular carcinoma (HCC).Methods:Clinical data of 45 patients with HCC who underwent anatomical hepatectomy in the Department of Hepatobiliary Surgery, the 900th Hospital of Joint Logistic Support Force of People′s Liberation Army from September 2019 to December 2020 were retrospectively analyzed. Among them, 27 patients were males and 18 were females, aged from 28 to 73 years, aged (57.76±10.95) years on average. According to the different surgical methods, all patients were randomly divided into ICG group ( n=24) and control group ( n=21). In ICG group, 15 patients were males and 9 females, aged (58.21±11.81) years on average. Anatomical hepatectomy was performed using 3D visualization combined with ICG fluorescence imaging. In control group, 12 patients were male and 9 female, aged (57.24±11.35) years on average. Conventional anatomical hepatectomy was performed. The operation duration, bleeding volume, the numbers of cases underwent blood transfusion, occlusion durations, days of hospitalization, highest postoperative serum ALT and TBiL, duration of recovery of postoperative serum ALT and TBiL, and incidence of complications were observed. Measurement data with approximately normal distribution were represented by ( Mean± SD) and groups were compared using t test. Measurement data with skewed or uneven distribution were represented by M (rang) and groups were compared using Man-Whitney U test. Count date were compared using Fisher exact test. Results:All the patients underwent successful operations, without perioperative death. In ICG group, the operation duration was 110.50 (44.00-145.00) min and the occlusion durations was (15.17±2.14) min respectively, shorter than 122.00(80.00-255.00) min and (17.29±4.35) min in control group, the difference between the two groups were statistically significant ( Z=-2.002, -2.115; P<0.05). In ICG group, the numbers of cases underwent blood transfusion was 2 cases, less than 8 cases in control group, the difference between the two groups was statistically significant ( χ2=4.147, P<0.05). The bleeding volume, days of hospitalization, highest postoperative serum ALT and TBiL, duration of recovery of postoperative serum ALT and TBiL, postoperative complications between the two groups were not statistically different ( P>0.05). Conclusions:3D visualization combined with indocyanine green fluorescence imaging technique is a feasible surgical method for anatomical hepatectomy. It is helpful for liver surgeons to visualize and recognize the boundary between hepatic segments and improve the safety of anatomic hepatectomy.

Objective To develop a simple high-resolution melting ( HRM) analysis method for differentiation of Pc and P2 variants in class 1 integron.Methods DNA fragments containing Pc and P2 variants were amplified from plasmids pACW ( PcW ) and pACWP2 ( PcW-P2 ) respectively , then these purified PCR products and P 2 promoters were analyed full-length amplicon by HRM .Eight DNA fragments containing different Pc promoters were amplified and site-specific mutated from plasmids pACS ( PcS ) , pACH2 ( PcH2 ) , pACH1 ( PcH1 ) , pACW ( PcW ) , genomic DNA of Klebsiellar pneumonia HS07-68 (PcWTGN-10)and HS05-1792(PcH2TGN-10)respectively.The purified PCR products and eight Pc variants were characterized by HRM analyses of an unlabeled probe and full-length amplicon.This assay was applied to the differentiate Pc and P2 variants in 109 class 1 integrons from 95 urine clinical Escherichia coli isolates in Huashan Hospital during 2004 -2007.The differentiation results were compared with that determined by direct sequencing .Results P2 promoter with a significant higher melting temperature ( Tm ) can be identified by HRM analysis clearly .P2 promoters were identified in 2 class 1 integrons and consistent with direct sequencing results .Eight Pc variants were classified into three groups: PcS, PcSTGN-10 , PcW, PcWTGN-10, PcH1, PcH1TGN-10.Using direct HRM analysis.PcH2, PcH2TGN-10 were classified into four groups:PcS, PcH1, PcH2, PcW, PcSTGN-10 , PcH1TGN-10 , PcH2TGN-10 , PcWTGN-10 according to the melting curves of the unlabeled probe .Combined the HRM analyses of the whole amplicon and unlabeled probe , the eight Pc variants can be differentiated from each other .Five different Pc variants, PcS, PcW, PcH1, PcH2TGN-10 and PcWTGN-10 , were identified and consistent with direct sequencing results .Conclusions This developed a simple Pc and P 2 variants differentiation method via simultaneous HRM analyses of an unlabeled probe and full-length amplicon .This method is cost-effective and accurate , could be used in differentiation of Pc and P2 variants of class 1 integrons in clinical isolates .

OBJECTIVE:To establish a method for simultaneous determination of tanshinol,caffeic acid,rosmarinic,salviano-lic acid B,salvianolic acid A,tanshinoneⅠ,cryptotanshinone,tanshinone ⅡA and ursolic acid in Compound xueshuantong cap-sules. METHODS:UPLC-MS/MS method was adopted. The determination was performed on ACQUITY UPLC? BEH C18 column with mobile phase consisted of acetonitrile-0.1%formic acid(gradient elution)at the flow rate of 0.2 mL/min. The column tempera-ture was 40 ℃,and the temperature of injector was 10 ℃. Analysis time was 7 min,and sample size was 5 μL. The electrospray ionization source(ESI)was used;ion source temperature was 150℃;capillary voltage was 3.5 kV;cone flow was 50 L/h;desol-vation temperature was 350 ℃;desolvation gas flow was 650 L/h;nebuliser pressure was 7 × 105 Pa;ion monitoring and multiple reaction monitoring (MRM) was performed. RESULTS:The linear ranges of tanshinol,caffeic acid,rosmarinic,salvianolic acid B,salvianolic acid A,tanshinoneⅠ,cryptotanshinone,tanshinone ⅡA and ursolic acid were 10.0-100.0 μg/mL (r=0.9998), 0.1-1.0 μg/mL(r=0.9998),4.0-40.0 μg/mL(r=0.9999),10.0-100.0 μg/mL(r=0.9999),15.0-150.0 μg/mL(r=0.9997), 8.0-80.0 μg/mL(r=0.9998),10.0-100.0 μg/mL(r=0.9997),50.0-500.0 μg/mL(r=0.9997)and 6.0-60.0 μg/mL(r=0.9998), respectively. The limits of quantitation were 40.0,9.6,38.0,88.0,130.0,39.0,4.4,3.2 and 10.0 ng/mL,separately. The limits of detection were 12.0,3.0,11.0,26.0,39.0,12.0,1.3,1.0 and 3.0 ng/mL,respectively. RSDs of precision,stability and repro-ducibility tests were all lower than 3%. The recoveries were 97.34%-103.20%(RSD=2.19%,n=6),97.22%-102.39%(RSD=2.03%,n=6),98.51%-101.70%(RSD=1.32%,n=6),97.86%-102.49%(RSD=2.09%,n=6),96.75%-103.12%(RSD=2.36%,n=6),98.43%-101.65%(RSD=1.25%,n=6), 97.59%-101.50%(RSD=1.50%,n=6), 96.45%-102.88%(RSD=2.58%,n=6),97.02%-103.11%(RSD=2.38%,n=6),separately. CONCLUSIONS:The method is simple and accurate,and can be used for simultaneous determination of 9 components in Compound xueshuantong capsules.

Objective To investigate the expression of miR-424* in 2 and 4 Gy X-ray irradiated A549 cells in vitro and in vivo,as well as in clinical lung tissues and serum sample of non-small cell lung cancer(NSCLC) patients,and to explore its potential role in the diagnosis and prognosis of lung cancer.Methods A549 cells were irradiated with 2 and 4 Gy X-rays,and some of irradiated cells were injected into nude mice through tail vein.Real time quantitative PCR (RT-qPCR) assay was employed to detect the expression of miR-424 * in 2 and 4 Gy X-ray irradiated A549 cells in vitro and in vivo,as well as in clinical lung tissues and serum sample of lung cancer patients.Results Compared with the control group,the expression of miR-424* was up-regulated significantly in X-ray irradiated A549 cells at 1,2,12,24 and 48 hpost irradiation,respectively (2 Gy:t =-45.886--6.709,P ＜0.05;4 Gy:t =-29.087--7.833,P ＜ 0.05).Furthermore,the expression of miR-424 * was up-regulated in the lung and serum of nude mice with injection of 0,2 and 4 Gy X-ray irradiated A549 cells,compared with control group (fold change was 9.72,8.58 and 4.7 with 2 Gy irradiation and 11.93,9.22 and 8.99 with 4 Gy irradiation,t=-13.243,-12.409,-9.833 in lung andt=-6.436,-3.052,-3.609 in serum,respectively,P ＜ 0.05).Out of 11 tissue samples of NSCLC patients,6 were detected with up-regulated miR-424* expression,and no significant discrepancy of miR-424* expression was detected in two type of NSCLC tissue samples.On the contrary,43 serum samples were detected with up-regulated miR-424* expression out of 84 serum samples (51.20％) of NSCLC patients (fold change range 1.97 to 17.71),and significant discrepancy of miR-424* expression was shown in two subtypes of NSCLC serum samples [adenocarcinoma:39.10％ (18/46) and squamous carcinoma:65.8％ (25/38)],as well as in serum samples of NSCLC patients with radiotherapy [41.5％ (22/53)] and without radiotherapy [67.7％ (21/31)] (t=5.919,5.387,P ＜0.05,respectively).Conclusions 2 and 4 Gy X-ray irradiation could up-regulate the expression of miR-424* in A549 cells,which might be correlated with the enhanced metastasis of A549 cells induced by X-ray in vivo and in vitro.Furthermore,the expression of miR-424* was up-regulated in over 50％ of the tissue and serum samples of NSCLC patients,which might be correlated with the diagnosis of NSCLC subtype and prognosis of radiotherapy.

OBJECTIVETo investigate the methylation in promtor region of RASSF2 and sFRP1 in sporadic colorectal cancer patients in order to provide screening method for early colorectal cancer.

METHODSThe methylation in promoter region of RASSF2 and sFRP1 in serum samples of 59 sporadic colorectal cancer patients and 59 healthy volunteers was detected by methylation specific PCR. The association between clinicopathological features of sporadic colorectal cancer patients and methylation in promoter region of RASSF2 and sFRP1 was analyzed.

RESULTSThe methylation rates of RASSF2 and sFRP1 gene in serum of 59 sporadic colorectal cancer patients were 27.1% and 30.5%, significantly higher than those in healthy volunteers(0%, both P<0.01). The methylation of RASSF2 or sFRP1 occurred in 29(49.2%) patients, which was significantly higer than the methylation rate of single gene(P<0.05). No association was found between methylation ratio of RASSF2 and sFRP1 and clinicopathological features in sporadic colorectal cancer patients.

CONCLUSIONSMethylation in promoter region of RASSF2 and sFRP1 is often detected in serum of colorectal cancer patients. The combination detection of methylation for the two genes may provide information for early screening of colorectal cancer.

Colorectal Neoplasms ; diagnosis ; genetics ; DNA Methylation ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; blood ; genetics ; Male ; Membrane Proteins ; blood ; genetics ; Middle Aged ; Promoter Regions, Genetic ; Tumor Suppressor Proteins ; blood ; genetics

CeO2-Co3 O4 composite nanofibers were prepared by the double jets electrospinning method. The nanofibers were then deposited onto the surface of a ω-type heating coil as cataluminescence material to prepare a new cataluminescence ( CTL ) type formaldehyde gas sensor. The crystalline phase and microstructure of CeO2-Co3 O4 composite nanofibers were characterized by X-ray diffraction ( XRD) and scanning electron microscope (SEM), and the cataluminescence mechanism and electrochemical characteristic of formaldehyde on the surface of CeO2-Co3 O4 nanofibers were analyzed by H2 temperature programmed reduction (H2-TPR) and X-ray photoelectron spectroscopy (XPS). Under the optimal conditions (500 nm of wavelength, 0. 2 L/ min of flow rate and 550℃), there was a good relationship between the CTL intensity of this formaldehyde gas sensor (Ce30) and formaldehyde concentration in the range of 1. 2 -50 μg / m3 , the sensitivity was 40. 04 a. u. / (μg / m3 ), the detection limit was 1. 2 μg / m3 , the dynamic response time and recover time of formaldehyde gas were 2. 4 s and 3. 5 s, respectively. The formaldehyde sensor was successfully applied to the determination of formaldehyde in the automotive exhaust, with relative errors of 0. 39% -1. 07% and relative standard deviations of less than 3% .

Objective:To probe the diagnostic performance of the combined evaluation of stenosis and plaque characteristics based on coronary computed tomography angiography (CCTA) in identification of myocardial ischemic lesions, using the invasive coronary angiography (ICA)-based fractional flow reserve (FFR) as the gold standard.Methods:From November 2018 to March 2020, the patients with suspected or known coronary artery disease and scheduled for ICA at 5 clinical trials centers were enrolled in this study. All the patients underwent CCTA, ICA and FFR in turn in one week. The luminal stenosis and plaque characteristics were measured and assessed including plaque burden, volume ratios of calcification and non-calcification, lesion length and CT vulnerable features. All culprit vessels were divided into FFR≤0.8 and FFR>0.8 groups, and the parameters of plaque characteristics were compared. The correlation of ischemic lesions with CCTA stenosis and plaque characteristics was analyzed by the logistic regression analysis. The ROC curve was used to evaluate the sensitivity and specificity of CCTA stenosis rate and plaque characteristics, meanwhile the area under curve (AUC) of each parameter was compared by Delong test.Results:Three hundred and sixty-six culprit vessels in 317 patients were analyzed in this study (169 vessels in ischemia group and 197 in nonischemia group). The plaque burden [34.3% (30.3%, 38.8%) vs. 32.4% (28.5%, 37.9%); Z=-2.622, P=0.009], proportion of CT vulnerable features [26.9% (45/169) vs.11.7% (23/197); χ 2=15.311, P<0.001] and lesion length [22.1 (14.4, 35.0) mm vs. 17.6 (11.0, 26.0) mm; Z=-4.388, P<0.001] in FFR≤0.8 group were higher than those in FFR>0.8 group. The results of logistic regression analysis revealed that CCTA stenosis, lesion length, and CT vulnerable features were significant predictors for myocardial ischemia (OR values: 3.794, 2.461, 1.027; P<0.001, P=0.002, P=0.002). The diagnostic performance of CCTA ≥50% stenosis alone in identification of ischemic lesions was low (AUC=0.625). When it combined high-risk plaque characteristics and lesion length, the AUC was improved to 0.714 with a statistical significance. Conclusions:CCTA stenosis, lesion length, and CT vulnerable features are major predictors in identification of myocardial ischemic lesions, and the combination will significantly improve the diagnostic performance of CCTA ≥50% stenosis.

Objective To investigate the effect of melatonin against cerebral ischemia reperfusion injury (CIRI) in mice and its mechanism.Methods Thirty male C57BL/6 mice were randomly divided into sham operation group,CIRI group,and melatonin treatment group (n =10 in each group).A middle cerebral artery occlusion model was induced by suture method.The degree of brain injury was evaluated by neurological function score,brain water content,and cerebral infarction volume.Western blot analysis was used to detect apoptosis-related proteins Bim,Bcl-2,and endoplasmic reticuhm stress-related molecules C/ EBP homologous protein (C/EBP) expression.Results Compared with the CIRI group,the neurological function score was significantly improved,the degree of cerebral edema was significantly reduced,and the volume of cerebral infarction was significantly reduced in the melatonin treatment group (all P ＜0.05).In addition,the expression of Bcl-2 was significantly up-regulated in the melatonin treatment group,and the expression of Bim and CHOP was significantly down-regulated (all P ＜ 0.05).Conclusion Melatonin may play an anti-CIRI role by regulating CHOP,and endoplasmic reticulum stress plays an important role in CIRI.

Hilar cholangiocarcinoma(HCCA) is a hotpot and a difficult point in the field of hepatobiliary surgery. HCCA is the most common type of cholangiocarcinoma and is characterized by atypical early clinical manifestations, rapid progression and poor prognosis. There is no specific marker for HCCA and its preoperative diagnosis and evaluation mainly relies on imaging examination. Surgical treatment is still the main treatment, but most patients have lost the opportunity of surgical resection by the time of treatment. In recent years, a large number of studies have been conducted on the diagnosis and treatment of HCCA at home and abroad, and the efficacy of HCCA has been improved. Perioperative management, including the selection of preoperative drainage and perioperative chemoradiotherapy and others, improved postoperative survival. Among them, the application of preoperative radiotherapy and chemotherapy in the field of liver transplantation has achieved quite good results. Targeted therapy and immunotherapy have provided new treatment methods for HCCA. This paper reviews the diagnosis and multimodal treatment of HCCA.

Objective:To evaluate the effects of pegylated interferon (Peg-IFN) alfa-2b combined with nucleotide analogues (NAs) on the recurrence of hepatitis B-related liver cancer after resection, and to explore the changes of HBsAg and HBV DNA in patients with chronic hepatitis B liver cancer during postoperative treatment.Methods:The prospective study was conducted. Clinical data of 43 patients with hepatitis B-related liver cancer who underwent radical resection treated in 900th Hospital of People′s Liberation Army were prospectively analyzed from January 2020 to December 2021. Among 43 patients, there were 39 males and 4 females, the age was 30-76 years. According to different treatment methods they were divided into two groups, the patients treated by Peg-IFN alfa-2b combined with NAs were devided into the IFN group( n=10), and those treated by NAs alone into the NAs group( n=33). Two-pair semi-quantitative were collected every 3 months after operation. The recurrence-free survival rate, recurrence time after 2 years in the two groups, the clearance rate and the negative rate of HBsAg and HBV DNA in the two groups. Peg-IFN alfa-2b was evaluated in improving the prognosis of hepatitis B-related liver cancer. The measurement data of normal distribution were expressed by mean±standard deviation ( ± s), and t-test was used for comparison between the two groups. Chi-square test was used for comparison between the two groups of count data. Repeated analysis of measurement variance was used for analysis HBsAg and HBV DNA changes of the interferon group overall survival time and recurrence-free surrival time of patients was estimated using Kaplan-Meier method and the difference between groups was assessed using Log-rank test. Results:HBsAg and HBV DNA: The HBsAg clearance rate at 24 weeks and that at 48 weeks in the IFN group were 24.6% and 59.0% respectively. The HBsAg negative rate at 48 weeks was 16.7%. The HBV DNA clearance rate at 24 weeks and that at 48 weeks were 33.9% and 53.8% respectively. The HBV DNA negative rate was 0 at 48 weeks. The levels of HBsAg and HBV DNA in the IFN group decreased gradually with time. There were statistically differences between the levels of HBsAg and HBV DNA at 0 weeks, 24 weeks and 48 weeks( P<0.05). The 2-year overall survival rates of IFN group and NAs group were 100% and 90.9% respectively. The 2-year recurrence-free survival rates were 90.0% and 63.6% respectively. There were no significant statistical differences in the overall survival rate and recurrence-free survival rate between the groups ( P>0.05). The postoperative recurrence time of the IFN group and the NAs group were (15.00±7.07) months and (5.78±3.39) months respectively. The difference between the two groups was statistically significant ( t=3.160, P<0.01). Conclusion:Long-term antiviral therapy of Peg-IFN alfa-2b combined with NAs can prolong the recurrence time of liver cancer, reduce the levels of HBsAg and HBV DNA in serum, and potentially improve the survival rate of the patients compared with therapy of NAs alone.