1.Aptamer-based Fluorescence Resonance Energy Transfer Method for Determination of Estradiol
Xiaohua JIANG ; Wenjie DING ; Chonglin LUAN
Chinese Journal of Analytical Chemistry 2014;(9):1295-1300
A highly sensitive fluorescence spectroscopic method was established for the selective determination of estradiol, which took advantages of the excellent molecular recognition capability of aptamer and the energy transfer between the specific fluorescent groups. The effects of the pH value, buffer constituent and concentration, the concentration of DNA, the experimental temperature and response time on the detection of estradiol were studied. Under the optimal conditions (50 mmol/L BR buffer solution with pH value at 7. 4, 1. 0×10-7 mol/L for each DNA strand, incubation at 45 ℃, response time 19 min), the change of the fluorescence intensity (ΔI) versus the logarithm of the concentration of estradiol ( lgC) was linear over a concentration range from 1. 0×10-11 mol/L to 5. 0×10-9 mol/L with good linear correlation (r=0. 9953). The limit of detection (LOD) was found to be 6. 0×10-12 mol/L (S/N=3). This method was successfully applied to the detection of estradiol in human urine, with the recovery in the range of 94. 0%-103. 5%. This method showed good precision and accuracy.
2.Construction expression and activity evaluation of a novel fusion toxin hIL15M/TNF?M
Fang LIU ; Lei DING ; Xiaohua MAO ;
Journal of Third Military Medical University 2003;0(08):-
Objective To construct and express fusion toxin human IL15 mutant/TNF? mutant (hIL15M/TNF?M) in order to develop a targeted toxin capable of eliminating abnormal cells expressing high levels of hIL15 receptors. Methods DNA fragments of truncated hIL15M and TNF?M coding gene were joined in frame and cloned into T7 expression vector pET16b. The expressed fusion protein was purified from E. coli . Cytotoxicity of hIL15M/TNF?M was measured by MTT assay. Results E. coli transformed with the recombinant plasmid expressed an extra band with the expected size of 30?10 3. MTT assay indicated that PHA activated T cells were 13 times more sensitive to the cytotoxic effect of hIL15M/TNF?M than to the resting T cells. Conclusion hIL15M/TNF?M has high toxic effect against cells expressing high levels of IL15 receptors, suggesting that hIL15M/TNF?M has potential therapeutic effect on T cell leukemia and other diseases related with improperly activated T cells.
3.Laparoscopic right hemihepatectomy following a standardized technique
Xiaohua YANG ; Ding SUN ; Zuxiong TANG ; Lei QIN ; Haixin QIAN
Chinese Journal of Hepatobiliary Surgery 2017;23(7):452-455
Objective To determine the clinical value of laparoscopic right hepatectomy (LRH) carried out following a standardized technique.Methods The medical records of 15 patients who underwent LRH at the First Affiliated Hospital of Soochow University were retrospectively reviewed.The perioperative indicators which included the operation duration,blood loss,tumor diameter,hospitalization duration and postoperative complications were analyzed.Results There was no conversion to open surgery.The operation duration was (251.1 ± 73.3) min.The resected tumor diameter was (8.5 ± 4.2) cm,the blood loss was (550.8 ± 343.6) ml,and the hospitalization duration was (10.2 ± 3.7) days.There was one patient who developed postoperative bile leakage.There was no other complication and there was no perioperative death.Conclusions The standardized technique of LRH was easy and effective.This technique shortened the operation duration and improved patient safety.
4.Pathogenic Bacteria in Urinary Tract Infection after Cesarean Section:Their Distribution and Drug Resistance
Jinlong DING ; Qunhua YING ; Li YANG ; Xiaohua SONG
Chinese Journal of Nosocomiology 2009;0(18):-
OBJECTIVE To understand the urinary tract infections after cesarean section,the distribution of pathogenic strains,and drug resistance analysis in order to provide clinical treatment. METHODS The routine urine cultures of the 2652 case were identified and drug resistance was analyzed by K-B method. RESULTS From 458 isolated bacteria,Gram-negative bacilli were main,pathogen accounting for 80.13%,Escherichia coli and Klebsiella,accounted for 71.62% and 4.8% respectively;Gram-positive bacteria accounted for 14.41%,and fungi accounted for 3.71%. CONCLUSIONS G-bacteria in the urinary tract infection are in the absolute superiority;rational use of antibiotics to effectively control the urinary tract infections and avoid drug-resistant strains produceing is especially critical.
5.Gene mutation and polymorphism analysis of human papillomavirus type 16 E6 in 98 cervical cancer samples
Xiaohua DING ; Hongbin CAI ; Fan ZHANG ; Yu LIANG ; Zhaoqiu YANG
Chinese Journal of Microbiology and Immunology 2008;28(11):989-992
Objective To study the mutation of papillomavirus type 16 (HPV16) E6 gene, and to compare with the result of other researchers. Methods DNA was extracted from cervical cancer tissue sam-pies, then HPV16 E6 genes were amplified and sequenced using the HPV16 E6 specific primers from cervi-cal cancer DNA samples. Results HPV16 E6 gene mutations at the position of nt178 were found in 18 case from 35 cervical cancer samples, the mutation rate was 51.43%, resulting in nucleotide change of Asp to Glu. E6 mutations are found at the position of nt442 in 4 cervical samples. E6 mutation was found in 1 to 2 case in other nucleotide. Conclusion High frequency mutations of E6 gene of HPV16 at the position of nt178, nt442 have been found in cervical cancer samples in Hubei, China. The role of E6 gene mutation in cervical cancer development should be studied further.
6.Role of toll receptor-4 in the increase of blood brain barrier permeability caused by acute pancreatitis
Zhen DING ; Jun LIU ; Rong LIN ; Wei QIAN ; Xiaohua HOU
Chinese Journal of Pancreatology 2009;9(2):112-114
Objective To investigate the relation between the expression of toll receptor-4 (TLR-4) and the increase of blood brain barrier (BBB) permeability in the rats with acute pancreatitis.Methods Male Sprague-Dawley rats were randomly divided into 8 groups:control group;acute edematous pancreatitis (AEP) 2 h,6 h groups;acute necrotizing pancreatitis (ANP) 2 h,6 h,12 h,24 h,48 h groups with 8 rats in each group.The BBB permeability,pathological score of pancreas,TLR-4 expression were determined and the relationships between them were analyzed.Results The BBB permeability in control group;acute edematous pancreatitis (AEP) 2 h,6 h groups;ANP 2 h,6 h,12 h,24 h,48 h groups were 1.55±0.29,1.64±0.17,1.69±0.24,1.89±0.12,2.66±0.32,2.91±0.29,2.89±0.69 and 1.84±0.07,respectively;the pathological scores of pancreas were 0,2.38±0.92,3.13±0.64,8.50±1.07,9.75±0.71,10.25±1.28,11.13±1.25 and 10.13±1.13,respectively;there was no significant difference between AEP groups and control group,while there was significant difference between AEP groups and ANP groups (P<0.05 orP <0.01).BBB permeability was correlated with pancreatic injury ( r = 0.626,P <0.01).There was no TLR-4 mRNA and protein expression in the control and AEP group,while they were significantly expressed in ANP groups,and the expression were positively related with BBB permeability ( r =0.208,P = 0.027 ).Conclusions BBB permeability was present in the course of ANP.Activation of TLR-4 signal pathway may be involved in the BBB permeability increase caused by ANP.
7.Relation between pleural effusion and severity of acute pancreatitis
Xiuying LU ; Jun HU ; Zhen DING ; Xiaohua HOU
Chinese Journal of Pancreatology 2009;9(5):309-311
Objective To investigate the incidence of pleural effusion in acute panereatitis(AP)and the relation between pleural effusion and severity of AP.Methods The medical records of 766 patients with AP were analyzed retrospectively.The incidence of pleural effusion was documented and its relationship with age of onset,sex,Ranson score,CTSI,serum albumin and hospital day was analyzed.Results In the 766 AP patients,there were 129(16.8%)patients had pleural effusion.In 171 SAP patients,there were 91 (15.3%)patients had pleural effusion.In the 595 MAP patients,there were 129(16.8%)patients had pleural effusion.The difference in the incidence of pleural effusion between MAP and SAP patients was statistically significant(P<0.05).The serum amylase,albumin,Ranson score,CTSI,and hospital day in patients with pleural effusion were(795±1013)U/L,(36±7)g/L,1.12±1.15,4.02±1.16 and(23.4±23.4)d,respectively;while the corresponding values in patients without pleural effusion were(592±856)U/L,(38±6)g/L,0.85±0.98,3.15±16.60 and(17.3±16.6)d,respectively.The difference in Ranson score,serum amylase,CTSI between the two groups was statistically significantly(P<0.05 or<0.01=,the serum level of albumin in patients with pleural effusion was significantly lower than that in patients without pleural effusion(P<0.01),the hospital day in patients with pleural effusion was significantly longer than that in patients without pleural effusion(P<0.01).Pleural effusion of both sides was present in 92 patients,while pleural effusion of right side was present in 11 patients,pleural effusion of left side was present in 26 patients;the site of pleural effusion was not associated with the severity of AP,however,patients with pleural effusion of left side tended to have longer hospital day(P<0.05).Conclusions Patients with pleural effusion during AP were more likely to be complicated with severe conditions,and the presence of pleural effusion may be a good marker for severity evaluation.
8.Imaging appearances of pulmonary mucosa-associated lymphoid tissue type lymphoma
Tiannü LI ; Qingjuan HUANG ; Chongyang DING ; Xiaohua ZHU ; Qiyong DING ; Jianwei CHEN
Chinese Journal of Radiology 2011;45(2):149-152
Objective To assess the value of CT or PET-CT with fluorine-18-labeled fluorodeoxyglucose (FDG) for the diagnosis of pulmonary mucosa-associated lymphoid tissue (MALT) type lymphoma. Methods The CT or FDG PET-CT findings in 14 patients with pathologically proved pulmonary MALT lymphoma were retrospectively analyzed. Results Lung lesions were unilateral in 7 patients and bilateral in 7 patients. Lesions presented as a single mass in 3 patients, as a single consolidation in 3 patients, as a nodule in 1 patient, as multiple nodules in 1 patient, as multiple patchy consolidations in 4 patients, as a mass with multiple nodules and patchy consolidations in 1 patients, as diffuse interstitial change in 1 patients. Air bronchogram was found in 9 patients and CT angiogram sign in 5 patients. On PET-CT, lesions showed heterogeneous FDG uptake in 2 patients, maximum standard uptake value was higher than 2. 5. Conclusion Imaging characteristics of pulmonary MALT lymphoma are single or multiple nodules or consolidations with air bronchogram on CT, and heterogeneous high FDG uptake on PET-CT.
9.The effect of anti-retroviral therapy on interleukin (IL)-7/IL-7R in HIV infected patients in China
Gang HE ; Peilin ZHEN ; Peipei DING ; Xiaohua CHEN ; Shaoqun LIANG ; Xingliu WU ; Changzheng HU
Journal of Chinese Physician 2014;16(3):330-332,335
Objective To explore the effect of anti-retroviral therapy on interleukin(IL)-7/IL-7R in human immunodeficiency virus(HIV) infected patients in China.Methods Cases were divided into 2 groups:HIV-infected group (35 cases),and control group (30 cases).IL-7 in serum,IL-7R(CD127) expression in CD4 +T cells,and CD4 +T cells count were detected and compared between two groups before and after treatment for 1 year.Results IL-7 level in the serum of HIV infected group before treatment [(8.98 ±3.77) pg/ml] was significantly higher than that in control group [(3.84 ±0.86) pg/ml] (P <0.05).The counts of CD4+T cells [(202.65 ± 121.54)/μl],CD4 + CD127 + T cells [(60.25 ± 11.75) %],and CD8 + CD127 + T cells [(46.27 ± 12.10)%] in HIV-infected group were significantly lower than those in control group [(766.99 ± 103.21)/L,(76.89 ± 20.01) %,(81.27 ± 12.35)%] (P <0.05).After anti-retroviral therapy (ART),IL-7 level in the serum of HIV-infected group[(5.55 ± 1.35) pg/ml]was decreased,and CD4+T cells [(450.58 ± 15)/μl],CD4 + CD127 +T cells [(69.82 ± 15.24)%],and [CD8 + CD127 + T(59.23± 14.73) %] cells was increased in HIV-infected group,with a significant difference between two groups (P <0.05).Conclusions ART could improve the IL-7 level in the serum and IL-7R(CD127)expression in CD4 +T cells of HIV-infected patients.However,they still cannot become normal level.
10.The infection status and genotype distribution of high risk human papillomavirus in Chongqing women
Haiwei ZHANG ; Ding LIN ; Changhai LIN ; Yu LIU ; Lixue CHEN ; Xiaohua ZHANG
Chongqing Medicine 2014;(36):4900-4902,4905
Objective To evaluate the infection status and genotype distribution of high risk human papillomavirus (HPV) in Chongqing women with different healthy status of cervix .Methods A retrospective analysis including 3 118 cases from June 2013 to March 2014 were performed ,945 of which were carried out multiplex real time PCR ,and data of cervix healthy status and HPV positive cases were subjected to chi‐square test .Results The general positive percentage of high risk HPV was 27 .49% ,16 .35%in healthy check‐up group ,and 24 .42% in cervicitis group and 39 .87% in cervical neoplasia lesion ,there was a significant difference of HPV positive ratio between these three groups(P< 0 .05) ;and HPV positive ratio was higher in older women without significant difference .In Chongqing region ,the three most common HPV types were HPV52 (21 .97% ) ,HPV16 (20 .00% ) and HPV58 (14 .75% ) .However ,the three most common HPV types in various groups are different :HPV52 (26 .20% ) ,HPV58 (14 .3% ) and HPV51 (10 .70% ) in health check‐up group ; HPV52 (21 .10% ) ,HPV58 (20 .00% ) and HPV16 (18 .90% ) in cervicitis group while HPV16 (29 .00% ) ,HPV52 (19 .80% ) and HPV58 (11 .50% ) in cervical neoplasia lesion group .The multiple infec‐tion rate of HPV was 5 .86% ,as the cervix status gets worse ,HPV concurrent infection increases ,and concurrent HPV types vary :mostly with HPV 52 in healthy check‐up group while with HPV 16 in cervical neoplasia lesion group .Conclusion The prevalence and genotype distribution of HPV in Chongqing women with different healthy status of cervix are significant difference.