Total Saponins in Paridis Rhizoma： A Review / 中国实验方剂学杂志

Paridis Rhizoma possesses the functions of clearing heat and detoxifying， alleviating swelling and relieving pain， cooling the liver and calming the convulsion. Saponins are the main active components of Paridis Rhizoma. Studies have shown that total saponins in Paridis Rhizoma have obvious inhibitory effect on solid tumors such as breast cancer， lung cancer， gastric cancer， and liver cancer and non-solid tumors such as leukemia. The saponins may exert the anti-tumor effects by inhibiting the proliferation， migration， and invasion of tumor cells， regulating cell cycle， inducing apoptotic and non-apoptotic death pathways， and regulating metabolism and tumor microenvironment. Furthermore， total saponins in Paridis Rhizoma showed anti-inflammatory， antioxidant， antimicrobial， hemostatic， and uterus-contracting activities. At the same time， they may induce apoptosis of normal cells， inflammation and oxidative stress， and metabolic disorders. In recent years， the reports of liver injury， reproductive injury， gastrointestinal injury， hemolysis， and other adverse reactions caused by total saponins in Paridis Rhizoma have been increasing. Pharmacokinetic studies have shown that there are significant differences in the metabolism of total saponins in Paridis Rhizoma administrated in different ways. Injection has a fast clearance rate， while oral administration may have hepatoenteric circulation. Meanwhile， due to the low solubility and activation of P-glycoprotein （P-gp） molecular pump， the prototype absorption， intestinal permeability， and recovery rate of total saponins in Paridis Rhizoma are poor， which affects the bioavailability. The bioavailability can be improved to some extent by preparing new dosage forms or new drug delivery systems with advanced technology. This paper reviews the pharmacological effect， pharmacokinetics， and adverse reactions of Rhizoma Paridis total saponins by searching the China National Knowledge Infrastructure （CNKI）， VIP， and Web of Science with ''Rhizoma Paridis total saponins'' as the keywords， hoping to provide references for the research， development， and clinical application of such components.

Diagnostic value of serum TIM-3 in patients with liver cancer / 国际肿瘤学杂志

Objective:To analyze the changes of T cell immunoglobulin- and mucin-domain-containing molecule-3 (TIM-3) in serum of patients with liver cancer and its diagnostic value.Methods:From March 2021 to May 2021, 37 patients with viral hepatitis type B (hepatitis B group) , 44 patients with liver cirrhosis (liver cirrhosis group) and 27 patients with liver cancer (liver cancer group) were selected in the Second Affiliated Hospital of Air Force Medical University, and 35 healthy subjects who underwent physical examination during the same period were selected as the healthy control group. The serum alpha fetoprotein (AFP) , liver function indexes and TIM-3 levels were detected, and the differences among groups were analyzed. The correlations between TIM-3 and AFP and liver function indexes were analyzed by Spearman correlation. Receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of TIM-3 in liver cancer.Results:There was a statistically significant difference in AFP among the hepatitis B group, liver cirrhosis group and liver cancer group ( χ2=11.75, P=0.003) . There were statistically significant differences in total bilirubin ( χ2=22.85, P<0.001) , direct bilirubin ( χ2=25.90, P<0.001) , indirect bilirubin ( χ2=19.92, P<0.001) , alanine aminotransferase ( χ2=36.64, P<0.001) , aspertate aminotransferase ( χ2=26.26, P<0.001) , aspertate aminotransferase/alanine aminotransferase ( χ2=34.67, P<0.001) and total bile acid ( χ2=13.10, P<0.001) among the hepatitis B group, liver cirrhosis group and liver cancer group. The serum levels of TIM-3 in the healthy control group, hepatitis B group, liver cirrhosis group and liver cancer group were 11.1 (4.2, 14.4) ng/ml, 12.7 (4.3, 23.9) ng/ml, 11.4 (3.4, 17.0) ng/ml and 15.7 (10.5, 21.2) ng/ml, with a statistically significant difference ( χ2=11.85, P=0.008) . There were statistically significant differences between the liver cancer group and healthy control group and liver cirrhosis group (both P<0.05) . Spearman correlation analysis showed that TIM-3 had no correlation with AFP in the four groups ( r=0.05, P=0.791; r=0.18, P=0.497; r=0.03, P=0.883; r=0.24, P=0.396) . There were correlations between serum TIM-3 and total protein in the healthy control group ( r=0.36, P=0.036) , serum TIM-3 and globulin in the hepatitis B group ( r=0.35, P=0.034) , and serum TIM-3 and total bile acid in the liver cancer group ( r=0.46, P=0.017) . ROC curve analysis showed that the sensitivity of serum TIM-3 for the diagnosis of liver cancer was 48.10%, and the specificity was 91.43%, when taking healthy subjects as the control group. The sensitivity of serum TIM-3 for the diagnosis of liver cancer was 96.30%, and the specificity was 41.77%, when taking healthy subjects and liver cirrhosis patients as the control group. The sensitivity of serum TIM-3 for the diagnosis of liver cancer was 96.30%, and the specificity was 40.52%, when taking healthy subjects, hepatitis B patients and liver cirrhosis patients as the control group. Conclusion:The serum level of TIM-3 in patients with liver cancer is significantly increased, which has certain diagnostic value for liver cancer, and can be used as a diagnostic marker and potential therapeutic target for liver cancer patients.

Detection of carcinoembryonic antigen levels in pleural effusion and serum and their ratio for differential diagnosis of pleural effusion resulting from tuberculosis and lung cancer / 南方医科大学学报

OBJECTIVE@#To study the clinical value of detecting carcinoembryonic antigen levels in pleural effusion (PCEA) and serum (SCEA) and their ratio (P/S) in the differential diagnosis of pleural effusions resulting from tuberculosis and lung cancer.@*METHODS@#This retrospectively study was conducted among 82 patients with pleural effusion caused by pulmonary tuberculous (TB; control group) and 120 patients with pleural effusion resulting from lung cancer in our hospital between April, 2016 and March, 2018. PCEA, SCEA and P/S were compared between the two groups and among the subgroups of lung cancer patients with squamous cell carcinoma (SqCa), adenocarcinoma (ACA), small cell carcinoma (SCLC). The receiveroperating characteristic curve (ROC) analysis was used to confirm the optimal critical value to evaluate the diagnostic efficiency of different combinations of PCEA, SCEA and P/S.@*RESULTS@#PCEA, SCEA and P/S were significantly higher in the overall cancer patients and in all the 3 subgroups of cancer patients than in the patients with TB ( < 0.05). The areas under the ROC curve of PCEA, SCEA and P/S were 0.925, 0.866 and 0.796, respectively; PCEA had the highest diagnostic value, whose diagnostic sensitivity, specificity, accurate rate, and diagnostic threshold were 83.33%, 96.34, 88.61%, and 3.26 ng/ml, respectively; SCEA had the lowest diagnostic performance; the diagnostic performance of P/S was between that of SCEA and PCEA, but its combination with SCEA greatly improved the diagnostic performance and reduced the rates of misdiagnosis and missed diagnosis. Parallel tests showed that the 3 indexes combined had significantly higher diagnostic sensitivity than each or any two of the single indexes ( < 0.05), but the diagnostic specificity did not differ significantly. The area under the ROC curve of combined detections of the 3 indexes was 0.941 for diagnosis of lung cancer-related pleural effusion, higher than those of any other combinations of the indexes.@*CONCLUSIONS@#The combined detection of PCEA, SCEA and P/S has a high sensitivity for diagnosis of lung cancer-related pleural effusion and provides important information for rapid and accurate diagnosis of suspected cases.

HER2 status in gastric adenocarcinoma of Chinese: a multicenter study of 40 842 patients / 中华病理学杂志

Objective@#To investigation HER2 status in gastric adenocarcinoma of Chinese and contributing factors to the HER2 expression. @*Methods@#HER2 status of 40 842 gastric adenocarcinomas and clinical data were retrospectively collected from 23 hospitals dated from 2013 to 2016. The association between HER2 positivity and clinicopathologic features was analyzed. @*Results@#Of the 40 842 patients the median age was 62 years, the male female ratio was 2.6∶1.0. The rate of HER2 positivity was 8.8% (3 577/40 842). HER2 expression was related to the tissue type, tumor location, Lauren classification and tumor differentiation (P values: 0.009, 0.001, <0.01 and <0.01, respectively). Different HER2 expression status was observed between primary and recurrent tumors in 7.6% (48/635) cases. The rates of HER2 positivity ranged from 2% to 10% among different institutions. The rates of HER2 FISH amplification were dramatically different among the 23 hospitals (0-100%) with an average rate of 10% (810/8 156) in patients with HER2 IHC 2+ . @*Conclusions@#HER2 expression is associated with clinicopathologic characteristics. HER2 re-assessment of tumor tissue and use of in situ hybridization techniques increase HER2 positivity. The current retrospective study should reflect the HER2 status in gastric adenocarcinoma of Chinese patients.

Applicable Value of AMSS-PCR in Lung Cancer Gene Mutation Detection / 中国肺癌杂志

BACKGROUND@#The detection of driver oncogenes of lung cancer is of great importance. There are various gene detection techniques nowadays which are different from each other. We carried out this study to investigate the specificity and sensitivity of assay panels based on an Amplification Refractory Mutation System-polymerase chain reaction (ARMS-PCR) technique of Amplification Mutation Specific System (AMSS) in detection of lung cancer gene mutation. To estimate the applicable value of assay panels in clinical settings.@*METHODS@#We collected cancer tissue specimens or fluid specimens from 309 patients. Mutation results were presented for those samples previously detected by ARMS-PCR. In comparison, we carried out AMSS-PCR using (epidermal growth factor receptor, EGFR) assay panel and Six-Alliance assay panel as well as Sanger sequencing. Software SPSS 22.0 (SPSS IBM) was used for statistical analysis.@*RESULTS@#The rates of consistency between the results by assay panels and Sanger sequencing or ARMS-PCR were 97.41% and 97.73%, respectively. Besides, EGFR assay panel had higher consistency rates with other detection methods than Six-Alliance assay panel. As for consistency test, the Kappa values of assay panels with Sanger sequencing, assay panels with ARMS-PCR, and ARMS-PCR with Sanger sequencing were 0.946, 0.953, and 0.913, respectively. The receiver operating characteristic curve (ROC) area under curve (AUC) of assay panels was 0.976 referring to Sanger sequencing, and 0.975 as ARMS-PCR was referred to.@*CONCLUSIONS@#AMSS-PCR can make an optimal cancer gene mutation detection method for clinical settings.

Prognostic value of American Joint Committee on Cancer-tumor regression grading combined with ypTN staging in patients with locally advanced rectal cancer / 中华放射肿瘤学杂志

Objective To investigate the prognostic value of American Joint Committee on Cancer-tumor regression grading ( AJCC-TRG) combined with ypTN stage in patients with locally advanced rectal cancer (LARC),who were treated with neoadjuvant chemoradiotherapy,and to identify the subgroups with the worst prognosis. Methods A total of 263 patients with LARC,including 176 males and 87 females,with a median age of 55 years,were admitted to Sun Yat-sen University Cancer Center from 2004 to 2012.All the patients received neoadjuvant chemoradiotherapy before surgery and underwent total mesorectal excision at 6 to 8 weeks after radiotherapy. All the surgical specimens were reevaluated according to the AJCC ( 7th edition)-TRG system and ypTN staging criteria. The prognostic prediction by TRG combined with ypTN was evaluated using survival analysis. The Kaplan-Meier method was used to calculate the rates of overall survival ( OS ) , disease-free survival ( DFS ) , local recurrence-free survival ( LRFS ) , and distant metastasis-free survival ( DMFS ) . The log-rank test was used for survival comparison and univariate prognostic analysis. Results The median follow-up was 601 months. The 5-year rates of OS, DFS, LRFS, and DMFS for all patients were 800%,750%,970%,and 810%,respectively. There were significant differences in OS, DFS,and DMFS between different ypT/TRG subgroups and different ypN/TRG subgroups (all P<005). ypT3-4/TRG 2-3 and ypN1-2/TRG 2-3 subgroups showed the worst prognosis. The 5-year rates of OS,DFS, and DMFS of the two subgroups were 669%/560%, 522%/414%, and 609%/460%, respectively. Conclusions A combination of AJCC-TRG system and ypTN staging can better predict the prognosis of LARC and identify the subgroups with the worst prognosis, which may provide a clinical guidance for postoperative individualized decision on adjuvant therapy for LARC.

Prognostic value of American Joint Committee on Cancer-tumor regression grading combined with ypTN staging in patients with locally advanced rectal cancer / 中华放射肿瘤学杂志

Objective To investigate the prognostic value of American Joint Committee on Cancer-tumor regression grading ( AJCC-TRG) combined with ypTN stage in patients with locally advanced rectal cancer (LARC),who were treated with neoadjuvant chemoradiotherapy,and to identify the subgroups with the worst prognosis. Methods A total of 263 patients with LARC,including 176 males and 87 females,with a median age of 55 years,were admitted to Sun Yat-sen University Cancer Center from 2004 to 2012.All the patients received neoadjuvant chemoradiotherapy before surgery and underwent total mesorectal excision at 6 to 8 weeks after radiotherapy. All the surgical specimens were reevaluated according to the AJCC ( 7th edition)-TRG system and ypTN staging criteria. The prognostic prediction by TRG combined with ypTN was evaluated using survival analysis. The Kaplan-Meier method was used to calculate the rates of overall survival ( OS ) , disease-free survival ( DFS ) , local recurrence-free survival ( LRFS ) , and distant metastasis-free survival ( DMFS ) . The log-rank test was used for survival comparison and univariate prognostic analysis. Results The median follow-up was 601 months. The 5-year rates of OS, DFS, LRFS, and DMFS for all patients were 800%,750%,970%,and 810%,respectively. There were significant differences in OS, DFS,and DMFS between different ypT/TRG subgroups and different ypN/TRG subgroups (all P<005). ypT3-4/TRG 2-3 and ypN1-2/TRG 2-3 subgroups showed the worst prognosis. The 5-year rates of OS,DFS, and DMFS of the two subgroups were 669%/560%, 522%/414%, and 609%/460%, respectively. Conclusions A combination of AJCC-TRG system and ypTN staging can better predict the prognosis of LARC and identify the subgroups with the worst prognosis, which may provide a clinical guidance for postoperative individualized decision on adjuvant therapy for LARC.

Balloon vertebroplasty reinforced by injectable calcium sulfate cement versus posterolateral fusion for treatment of thoracolumbar burst fractures / 中国综合临床

Objective To compare the clinical curative effects of balloon vertebroplasty reinforced by injectable calcium sulfate cement(CSC) and posterolateral fusion after short-segment pedicle screw fixation through the fractured vertebra in treatment of thoracolumbar bumt fractures.Methods From February 2012 to May 2015,68 cases patients with single thoracolumbar burst fracture were admitted in Orthopaedics and Traumatology Department of Traditional Chinese Medicine Hospital of Gaozhou,and were randomly divided into two observation group and control group,each group with 34 cases.Observation group were treated by short-segment pedicle screw fixation through the fractured vertebra followed by transpedicular balloon reduction of the endplate and CSC injection,the control group were underwent posterolateral fusion in addition to short-segment pedicle screw fixation through the fractured vertebra.The 2 groups were compared at preoperation,and the last follow-up in terms of visual analog pain score(VAS),Oswestry disability index(ODI),anterior height ratio of the injured vertebra,middle height ratio of the injured vertebra,the vertebral wedge angle and the sagittal Cobb angle.Results The patients were followed up from 12 to 54 months.All the measure indexes at the last follow-up were significantly improved than those at preoperation in all the patients(P0.05).At the last follow-up,the observation group showed a significantly less loss than the control group regarding the corrections of anterior height ratio of the injured vertebra((93.6±3.4) vs.(81.8±4.8)),middle height ratio of the injured vertebra((90.7±3.8) vs.(78.9±5.1)),the veaebral wedge angle((5.7±1.9) ° vs.(8.8±3.0) °) and the sagittal Cobb angle((5.2±2.1) ° vs.(7.9±2.4) °)(t=4.143,5.786,4.819,4.465,P<0.05).Conclusion In treatment of thoracolumbar burst fractures,compared with posterolateral fusion,balloon vertebroplasty reinforced by injectable CSC can better restore the shape of injured vertebral body and stability of the anterior and middle columns in the early period,can also reduce losses of corrections,and keep the function of the spinal motion segment.

Progress on the Study and Application of Serum Adenosine Deaminase in the Diagnosis of Autoimmune Diseases / 现代检验医学杂志

Adenosine deaminase (ADA)plays an important role in the regulation of normal immune system.The altered activ-ity of serum ADA could associate with some autoimmune diseases(AID).Various studies abroad have found the elevated ac-tivity of serum ADA compared with healthy controls in many kinds of AID such as systemic lupus erythematosus (SLE), rheumatoid arthritis(RA),et al (P<0.05).Many studies foused on the correlation of altered activity of serum ADA with the disease severity,but there is still controversy.To explore the diagnostic value of serum ADA in AID,make a review a-bout the application research progress of ADA in several AID.

Evaluation of neutralizing antibodies in serum immunized with virus-like particle chimerized HCV series of neutralizing epitopes / 中国免疫学杂志

Objective:New Zealand rabbits were immunized with VLPs-MEpS,VLPs-E2S,and the levels of neutralizing antibodies in serum were determined.Methods: New Zealand rabbits were immunized with 10 μg VLPs-MEpS and VLPs-E2S,serum was collected at diffferent time with a two-weeks interval.The neutralizing antibodies were determined by ELISA.HCV(type 1b) had been prepared and mixed with serum from immunized rabbit before infected Huh7.5 cell.The protection of neutralizing antibodies in serum was assessed.Results: Neutralizing antibodies had been induced in rabbit after immunized with VLPs-MEpS and VLPs-E2S.VLPs-MEpS group had higher titer of antibodies than that of VLPs-E2S group(P<0.05),both group had higher titer of antibodies than that of control groups significantly(P<0.01).VLPs-MEpS group had higher neutralization than that of VLPs-E2S group(P<0.05),the highest neutralization rate was 61.49%.Both groups were higher than control group notably(P<0.01).Conclusion: Protective neutralizing antibodies have been induced in New Zealand rabbit after immunized with VLPs-MEpS and VLPs-E2S.It′s the basement for development of neutralizing antibodies vaccine.

Expression and purification of Trop2 virus like particles / 国际检验医学杂志

Objective To gain the virus like particles (VLPs) based on Trop2 targets ,which provided the basis for further in vi‐vo induced experiments and anti‐tumor vaccine studies .Methods Using molecular cloning method to constract eukaryotic expres‐sion vector of pCAGGs/Trop2 and baculovirus expression vector of pFastbac1/Trop2 ,expression product of pCAGGs/Trop2 in HeLa cells were intended to be to be anchored to the cell membrane by the methods of Immunohistochemistry ;pFastbac1/Trop2 were transformed into E .coliDH10bac isolates to gain recombinant bacmids ,which were transfected into insect cells to express re‐combinant baculovirus with Gag rBV ,then sucrose density gradient centrifugation ,Western Blot and electron microscope were per‐formed to purify and identify the Trop2 VLPs .Results Building recombinant bacmids which were transfected into insect cells to express recombinant baculovirus with Gag rBV ,gained the recombinant Virus like Particles .Conclusion Trop2 VLPs was success‐fully prepared ,which laid the foundation for the subsequent induction of humoral and cellular immune response .

Application of mouse anti AEG-1 monoclonal fluorescent antibody in malignant serous cavity effusions / 国际检验医学杂志

Objective To use mouse astrocytes elevated gene‐1 (AEG‐1) monoclonal fluorescent antibody for detecting tumor cells in malignant serous cavity effusions .Methods The expression of AEG‐1 in serous cavity effusion exfoliated cells by PCR and Western‐blot ;the mouse monoclonal anti‐AEG‐1 fluorescent antibody and tumor cells in malignant serous cavity effusions were co ‐incubated ,meanwhile ,the serous cavity effusions in benign lesions were taken as the negative control .Results The AEG‐1 expres‐sion was positive in malignant serous cavity effusions exfoliated cells ,while which in benign lesion serous cavity effusion was nega‐tive or weakly positive ;meanwhile the results of direct labelling in mouse anti AEG‐1 monoclonal antibodies were consistent with the results by PCR and Western‐blot .Conclusion Mouse anti AEG‐1 monoclonal fluorescence antibody can provide certain theoret‐ical basis for the detection of tumor cells in serous cavity effusion .

Construction and identification of Stathmin gene Pichia pastoris expression system / 国际检验医学杂志

Objective To provide the experimental basis for the further research of the interacting proteins with Stathmin ,the Stathmin gene Pichia pastoris expression system was constructed ,the expressed Stathmin product was purified and identified .Meth‐ods Stathmin gene was amplified from tumor cell line of SKBR3 by PCR method and cloned into the yeast expression vector pPIC3 .5K .The recombinant vector pPIC3 .5K‐Stathmin was constructed and transformed into Pichia pastoris GS115 .The positive clones were screened by YPD medium containing Geneticin 600 μg/mL .Expression was induced with 0 .5% methanol and expres‐sion products were identified by SDS‐PAGE and Western Blotting .Results DNA sequencing result showed that the gene fragment was consistent with Stathmin gene sequence .pPIC3 .5K‐Stathmin was selected from YPD culture medium containing Geneticin ,and the positive clones were identified by PCR .SDS‐PAGE showed that a 37 × 103 protein band could be seen on the PAGE gel after Coomassie Blue staining ,which was further confirmed and identified as Stathmin protein by Western Blotting .Conclusion Stathmin yeast expression vector is successfully constructed and expressed in Pichia pastoris ,which laid the foundation for the study of inter‐acting proteins with Stathmin ,and for the preparation of the biological treatment drugs of Stahtmin target .

Influence of injection cefotiam on vanadate oxidation method and dry chemical method assay total bilirubin / 国际检验医学杂志

Objective Discuss the interference of injection cefotiam on vanadate oxidation method and dry chemical method assay total bilirubin .Methods Collected 60 examples ,include total bilirubin concentration 20 examples less than 20 μmol/L ,20 examples between 150-220 μmol/L and 20 examples between 350-410 μmol/L ,add an equal volume of various concentrations of cefotiam in each case ,formulated into cefotiam final concentrations of 300 ,150 ,75 mg/L of serum samples as the test group ,add an equal volume of water in each serum samples as the control group ,determine all the samples total bilirubin concentration respectively by vanadate oxidation method and dry chemical method ,compared the interference of cefotiam on determined total bilirubin by two method ,analyze the data by SPSS13 .0 .Results Determined total bilirubin by dry chemical method ,the test group higher than the control group ,the difference was statistically significant(P<0 .05) ,at the same total bilirubin levels ,with cefotiam concentrations decreased ,increased rate of total bilirubin concentration were decreased in the experimental group .Determined total bilirubin by vanadate oxidation method ,when the total bilirubin concentration between 150 -220 μmol/L ,the test group was higher than the control group ,the difference was statistically significant(P<0 .05) .Conclusion Interference of injection cefotiam on determined to‐tal bilirubin by dry chemical method is strong ,and with the drug concentration increased ,effect is more obvious ,but determination of total bilirubin by vanadate oxidation method has almost no effect .

Correlation of Breast Cancer Cells Invasive and Stathmin Gene Expression / 现代检验医学杂志

Objective To explore the relationship between the Stathmin gene expression in breast cancer cells MDA-MB-231, MCF-7 and the biological behaviours such as cell growth,adhesion and invasion,and provide experimental basis of breast cancer metastasis for further study.Methods Used RT-PCR and Western Blot methods to detect the Stathmin gene expres-sion levels in MDA-MB-231 and MCF-7 cells,and in the mean while to test the MDA-MB-231 and MCF-7 cell growth,adhe-sion,invasion ability by CCK-8 cell proliferation experiments,cell adhesion experiments,cell invasion experiments,then, analyed the relationship of Stathmin gene expression and cell growth,adhesion,invasion ability.Results Over-expression levels of Stathmin gene were observed both in the MDA-MB-231 and MCF-7 cells (F=10.173,P<0.05),and furthermore, the expression levels of Stathmin gene in MDA-MB-231 cells was higher than in MCF-7 cells (t=4.562,P<0.05).While, the growth,adhesion and invasion ability of the MDA-MB-231 cells was higher than that of MCF-7 cells(P<0.05).Conclu-sion The higher level of Stathmin gene expression,the stronger breast cancer cells had ability of growth,invasion,and ad-hesive.The Stathmin gene expression levels was closely correlated with breast cancer cell invasive.

Prokaryotic expression and purification of anti-AEG-1 single-chain variable antibody / 国际检验医学杂志

Objective To construct anti-astrocyte elevated gene-1(AEG-1)single-chain variable antibody (V23)prokaryotic ex-pression vector,and to conduct the protein purification and immunological activity detection.Methods The Primer5 software was applied to design the primers aiming at the gene sequence of the antibody anti-AEG-1 single-chain variable region for constructing the prokaryotic expression plasmid of PRsetC/V23.After the enzyme digestion by the restriction enzyme Pst1 and correct DNA se-quencing,the prokaryotic expression plasmid was led to E.coli BL21 ,the prokaryotic expression engineering strain containing the V23 gene was constructed.After the induction with IPTG,the interest protein was purified by the magnetic beads with the HIS tag,and the content of interest protein was determined by the SDS-PAGE electrophoresis.Western blot and ELISA were adopted to detect the immune activity of the nti-AEG-1 single-chain variable region antibody.Results For the constructed prokaryotic expres-sion plasmid PRsetC/V23,the single enzyme digestion and sequencing analysis displayed that the constructed V23 gene was com-pletely consistent to the designing sequences.After IPTG induction,SDS-PAGE electrophoresis showed an apparent protein band at 31×103 ,the Western blot detection showed a specific AEG-1 response band at 80 ×103 ,the ELISA test showed the positive re-sults.Conclusion The PRsetC/V23 prokaryotic expression plasmid and the V23 prokaryotic expression engineering strain are suc-cessfully constructed,this engineering strain can express anti-AEG-1 single-chain variable region antibody protein,and the protein has good immune activity.

Study of preparation of the Annexin V-nanoscale ultrasound contrast agents and targeting ultrasound imaging in vitro / 中华超声影像学杂志

Objective To research the Annexin V-nanoscale ultrasound contrast agents'preparation, ultrasound imaging and the ability to binding apoptosis cells of tumor in vitro.Methods The nanoscale bubble (Nanobubbles,NBs ) packaged the octaflouropropane (C3 F8 ) gas was prepared by thin film hydration.The Annexin V-Nanobubbles (AVNBs ) solutions was acquired through conjugating the biotinylated-Annexin V to the surface of the NBs by biotin-streptavidin bridging chemistry.The size and zeta potential of AVNBs were measured by NanoPlus-3 zeta/nano particle analyzer.The shift in size distribution of AVNBs bubbles was analyzed for the stability,after it was stored at 4 ℃ for different time. AVNB's shape were measured by scanning electron microscopy.The AVNBs bubble was measured using an ultrasound system for echogenicity in vitro,and SonoVue was for control.Finally,the ability of AVNBs binding with apoptosis cells of tumor in vitro was determine via the fluorescence microscope.Results AVNBs has a size distribution of (640.2±32.1 )nm,and a mean zeta potential of (-23.30 ±5.71 )mV.Its size remained relatively constant and appeared to show less size variation within the 24 h analysis period. AVNBs solutions were visible milky white and slightly suspension liquid with the naked eye.Under scanning electron microscopy,AVNBs were uniform hollow sperical cavitation bubble with small size and larger dispersibility in solution.The AVNBs and SonoVue solution had the same higher grayscale signal intensity by ultrasonic imaging.The AVNBs binded well with apoptosis cells of tumor in vitro,and the rate of binding was (97.55 ± 1 .30 )%.Conclusions The AVNBs particles prepared by method of thin film hydration have a nanoscale size,good stability and echogenicity.It can be targeted binding with the apoptosis cells of tumor in vitro.

Comparability of 5 items of hepatitis B in ELISA by using water bath box and rapid incubation / 国际检验医学杂志

Objective To investigate and evaluate the comparison results of five items of hepatitis B in ELISA by using water bath box and rapid incubation .Methods 92 patients with hepatitis B in our hospital from June 2012 to June 2013 were randomly se‐lected and 184 cases of healthy physical examination were randomly selected as the controls .The water bath and rapid incubation were adopted to conduct ELISA for simultaneously detecting 5 items of hepatitis B in samples .Results The results of two groups had higher positive coincidence rate .The average OD value of specimens in the water bath was higher .But the specimen S/CO value in the fast average incubation was higher ,its co value was higher than that of the water bath .Conclusion The rapid incubation can be introduced for improving the work efficiency in the daily work ,but the specimen with weak reactiveresult needs to be re‐de‐tected by using the water bath and making records .The fast incubation device is best for only use of the HBsAg reaction ,because the other four reaction process time is shorter ,and there are some non‐conformity results .

Clinical significance of serum Cys-C,β2-MG and RBP detection in hemorrhagic fever with renal syndrome / 国际检验医学杂志

Objective To study the changes condition and clinical significance of serum cystatin C(Cys‐C) ,β2 microglobulin(β2‐MG) and retinol binding protein (RBP) detection in different stages of hemorrhagic fever with renal syndrome(HFRS) .Methods The levels of serum Cys‐C ,β2‐MG and RBP were detected in 22 patients with HFRS and 30 cases of healthy physical examination and the detection results were statistically analyzed by the SPSS13 .0 software .Results The serum Cys C and β2‐MG levels in every stage of HFRS were increased compared with the healthy control group(P0 .05) ,while RBP was significantly increased in the oliguria stage ,polyuria stage and convalescence stage(P 0 .05) .Conclusion β2‐MG is more sensitive than Cys‐C in the early stage of HFRS ;RBP has the clinical guidance significance in the progression of HFRS .

Recent curative effect evaluation of double Endobutton technique and clavicular hook plate in patients with Tossy grade Ⅲ acromioclavicular separation / 中国医师进修杂志

Objective To compare recent curative effect between double Endobutton technique and clavicular hook plate (CHP) in patients with Tossy grade Ⅲ acromioclavicular separation.Methods One hundred and forty-nine patients with Tossy grade Ⅲ acromioclavicular dislocation were divided into control group (78 cases,treated with CHP) and observation group (71 cases,treated with double Endobutton technique) according to treatment method.The postoperative injured shoulder visual analogue score (VAS),abduction-rise activity and flexion-rise activity,Constant-Murley score and Karlsson grade were compared between 2 groups.Results All the patients successfully completed the operation,and the postoperative follow-up time was 9-40 (15.8 ± 5.2) months.The injured shoulder abduction-rise activity and flexion-rise activity postoperative 9 months in observation group were (100.9 ± 13.3)° and (131.6 ± 13.4)°,in control group were (81.4 ± 8.4)° and (96.7 ± 10.9)°; the injured shoulder VAS in observation group and control group were (2.2-± 0.8) and (3.0 ± 0.9) scores,and there were statistical differences between 2 groups (P ＜ 0.01).The Constant-Murley score in observation group and control group were (85.4 ± 6.4) and (82.5 ± 6.2) scores,the excellent and good rate of Karlsson grade in observation group and control group were 84.5％(60/71),91.0％ (71/78),and there were no statistical differences (P ＞ 0.05).Conclusions The recent curative effect of double Endobutton technique and CHP in patients with Tossy grade Ⅲ acromioclavicular separation is similar.