1.Expression of human BTC with biological activity
han-bei, CHEN ; yan, DONG ; qing, SU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(02):-
Objective To obtain abundant human betacellulin(BTC) with biological activity. Methods The whole mature protein coding sequence of BTC gene was amplified by polymerase chain reaction(PCR) method applied to human pancreatic ?-cell tumors cDNA.The fragment was cloned into prokaryotic expression vector pET32a(+) plasmid.The recombinant plasmid was transformed into E.coli BL21 and the fusion protein was expressed under isopropyl-beta-D-thiogalactopyranoside(IPTG).The fusion protein was purified by Ni2+ affinity chromatography.SDS-PAGE and Western blot were employed to determine the expression and purification of the expected protein.BTC was added to culture NIH3T3 cells for 5 days,and cell proliferation was detected by MTT. Results Lots of fusion protein were produced,and the purified protein can stimulate the proliferation of NIH3T3 cells. Conclusion The human BTC can be successfully obtained from the pET32a(+) system with the biological activity of stimulating the proliferation of NIH3T3 cells.
2.Construction and expression of human PDX-1 eukaryotic expression vector
han-bei, CHEN ; yan, DONG ; qing, SU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(01):-
Objective To construct eukaryotic expression vector of human pancreatic duodenal homeobox 1(PDX-1) gene,and to detect its expression in NIH3T3 cell lines. Methods The whole coding sequence of PDX-1 gene was amplified by polymerase chain reaction(PCR) from human pancreatic-cell tumors cDNA.The fragment was inserted into eukaryotic expression vector pcDNA3.1 plasmid.The recombinant plasmid was verified by double digestion and DNA sequencing.The expression of PDX-1 gene in NIH3T3 cells was assayed by Western blot. Results The length of specific fragment amplified by PCR was 852 bp,and the recombinant plasmid pcDNA3.1-PDX-1 showed two bands of 5.5 kb and 852 bp by digestion using respective restriction enzymes BamHⅠand EcoRⅠ.The sequence of PDX-1 gene was approved or confirmed by blasting to GenBank.It was suggested that PDX-1 gene had been cloned into pcDNA3.1 vector correctly.Western blot showed that PDX-1 gene was expressed,which was detected 24 h after pcDNA3.1-PDX-1 plasmid was transfected into NIH3T3 cells. Conclusion The recombinant eukaryotic expression vector pcDNA3.1-PDX-1 was successfully constructed and expressed in NIH3T3 cell lines.
3.Clinical and imaging efficacy observation of closed reduction and plaster splint fixation in the treatment of ge-rontal patients with distal radius fractures
Daojun LU ; Caifeng ZHANG ; Dong HAN ; Zhifeng HAN ; Xi HAN ; Yan CHEN ; Zhong PENG
Chinese Journal of Primary Medicine and Pharmacy 2014;(5):677-678
Objective To discuss the clinical efficacy of closed reduction and plaster splint fixation ( CRPSF) in the treatment of gerontal patients with distal radius fractures ( DRF) .Methods 76 elderly patients with DRF who treated by CRPSF were selected .According to AO classification of fractures ,the patients were divided into three groups,the A group had 27 cases;B group had 26 cases,C group had 23 cases.The treatment effect was evalua-ted by analyzing the follow-up data,the corresponding imaging measurement parameters and clinical scores .Results All patients had 12-month clinical and imaging follow-up.In the last follow-up, the arm-shoulder-hand dysfunction score and palmar angle ,ulnar deviation angle of A and B group were significantly better than those of C group , the differences were statistically significant(all P<0.05),the difference between A and B group was not statistically sig-nificant (P>0.05).In the last follow-up,the satisfaction score of C group was slightly lower than that of the A or B group,but had no statistically significant difference (P>0.05).Conclusion CRPSF in the treatment of gerontal pa-tients with DRF has good function and imaging effects ,and the improvement level has a certain relationship with the degree of fracture,but has no significant impact on the patients'satisfaction.
4.Satge I clinical study of dose escalation of capecitabine during intensity modulated radiotherapy concurrent chemotherapy for local regional advanced nasopharyngeal carcinoma
Baomin ZHENG ; Yan SUN ; Shukui HAN ; Xiaoxia DONG ; Bo XU
Chinese Journal of Radiological Medicine and Protection 2010;30(3):330-332
Objective To decrease radiation induced toxicities especially mucostis in patients with locally advanced nasopharyngeal carcinoma( NPC ) who underwent concurrent radiochemotherapy, the maximum tolerated dose and dose limited toxicities of capecitabine combination with cisplatin were observed. Methods From Aug 2006 to Oct 2007, 24 patients with intensity modulated radiotherapy(IMRT) and concurrent chemotherapy with capecitabine and cisplatin for nasopharyngeal carcinoma(stages Ⅲ-Ⅳ) were enrolled in this study. There were four dose-level groups of Capecitabine[625-1250 mg/(m2 ·d) , d1-14]and fixed cisplatin dose[20 mg/(m ·d) ,d1-5) ]MRI and CT scan were used for evaluation of tumor shrinkage. Treatment related toxicities were evaluated according to the common toxicity criteria( NCI-CTC Version 3.0). Results The acute side-effects include Grade 3 or Grade 4 mucosal toxicity(lasting for at least 5 d) and Grade 3 or Grade 4 non-mucosal toxicity were evaluated. Group 625 mg/m2 and Group 825 mg/m2 had none, Group 1000 mg/m2 had 6 patients and Group 1250 mg/m2 had 3 patients for mucosal toxicity, which were the main dose-limited toxicity and relevant to the dose of capecitabine apparently( P < 0. 05 ). There was also a trend of increase by the dose level of capecitabine for other toxicities. The median follow-up time for all patients was 28. 5 months. The locoregional recurrence occurred in 2 patients and distant metastasis in 2 patients. Two-year overall survival rate and locoregional control rate were 100% and 91.7%, respectively.Complete response and partialresponse were found on MRI or CT scan in patients of 29. 2% at the end of treatment and 83. 3% after three months, respectively. Conclusions The combination regimen of capecitabine and cisplatin is safe and effective according to the preliminary result. Toxicities related to radiochemotherapy for NPC were significantly associated with the dose level of chemotherapy.
5.Evaluation of the application value of video-electroencephalogram monitoring in the diagnosis and localization of pediatric epilepsy
Qing LIU ; Yan DONG ; Hongwei ZHANG ; Yueming HAN
Chinese Journal of Postgraduates of Medicine 2021;44(6):519-523
Objective:To explore the application value of video-electroencephalogram(VEEG) monitoring in the diagnosis and localization of children with epilepsy.Methods:The clinical data of 310 children with clinical epilepsy symptoms diagnosed and treated in Tangshan Maternal and Child Health Care Hospital and Qilu Children′s Hospital Affiliated to Shandong University from May 2018 to April 2020 were analyzed. All children underwent routine EEG (REEG), VEEG monitoring, and positron emission computed tomography (PET-CT) for children undergoing surgery. REEG and VEEG was compared to monitor epileptic form discharges; the location of epileptic lesions was compared by different examination methods, and children were followed up after surgery.Results:Among the 310 children, 247 children were diagnosed with epilepsy and 63 children were suspected of epilepsy. The detection rate of VEEG epileptic form discharge was higher than that of REEG: 87.4% (216/247) vs. 45.7% (113/247), and the difference was statistically significant ( χ2 = 6.3042, P<0.05). Among 247 children with epilepsy, during the VEEG test, 81 children (32.8%) had clinical symptoms, of whom 65 had seizures and 16 had non-epileptic seizures. The detection rate of VEEG for epilepsy was higher than that of clinical seizures: 87.5% (216/247) vs. 32.8% (81/247), and the difference was statistically significant ( χ2 = 8.6148, P<0.05). Among the 247 children with epilepsy diagnosed, 144 children had epilepsy syndrome, temporal lobe epilepsy accounted for 54.17% (78/144), and frontal lobe epilepsy accounted for 34.2% (50/144). The accuracy of preoperative VEEG localization of epilepsy lesions in 50 children undergoing surgery was higher than that of PET-CT and REEG: 80.0% (40/50) vs. 56.0% (28/50), 54.0% (27/50), and the differences were statistical significance ( χ2 = 5.3014, 5.6031, P<0.05). Postoperative follow-up of 50 children who had underwent surgery showed that 50.0% (25/50) of the children had no seizures, and 38.0% (19/50) of the children had significant improvements in postoperative seizure time, seizure period, and clinical manifestations. 12.0% (6/50) of the children had no significant improvement in clinical symptoms. Conclusions:VEEG can improve the detection rate of epileptic form discharges, and provide an important basis for clinical diagnosis of epilepsy, and have important value in locating epilepsy lesions before surgery.
6."Effects of physical therapy guided by "" international classification of functioning,disability and health""on motor function of stroke patients"
Dong HAN ; Jinyan WANG ; Lihuizi SUN ; Jiayi XIA ; Yan ZHU
Clinical Medicine of China 2016;32(7):577-581
Objective To discuss how to use the core elements of international classification of functioning,disability and health(ICF) as a guide in stroke operation treatment to develop targeted,effective therapy plan and rehabilitation goals,establish individualized treatment of rehabilitation,and to observe the effect of limb movement function in stroke patients. Methods Sixty cases patients with stroke and aged from 40 to 65 years old were selected. The body function and structure of all stroke patients were marked by comprehensive evaluation scores of The National Institutes of Health Stroke Scale ( NIHSS ) and Fugl?Meyer assessment. And then they were randomly divided into two groups,30 cases in each group. The observation group used the Chinese version of brief ICF Core Sets for evaluation, and set individualized treatment according to the results of the assessment;The control group, according to the conventional rehabilitation process, involved in training routine physical therapy. Again 8 weeks after treatment for patients with evaluation,the assessment results was compared with the results of the first assessment. Results Before treatment, the NIHSS, Fugl?Meyer assessment and Chinese version of brief ICF core elements scale scores for the observation group were 7. 95±2. 37,34. 5±4. 0 and 82. 15±17. 96,for the control group were 7. 58±2. 25,34. 8±4. 0 and 81. 55±18. 78. After treatment,the above three rating scale scores, for the observation group were improved to 4. 78 ± 1. 63, 56. 3 ± 4. 2, 60. 45 ±11. 04,for the control group were improved to 5. 13±1. 75,48. 1±3. 8,73. 56±16. 89. The two groups were significantly improved ( observation group:t=4. 94, 15. 51, 18. 59, P<0. 05;control group:t=3. 17, 14. 66, 12. 33,P<0. 05). After treatment,compared with the control group of three rating scale scores(5. 13±1. 75,48. 1
±3. 8 and 73. 56±16. 89),the observation group(4. 78±1. 63,56. 3±4. 2 and 60. 45±11. 04) improved more apparently(t=3. 15,14. 54,15. 45,P<0. 05) . Conclusion Guided by the idea of ICF and scale,with the demand of the patients as the center,to develop personalized physical therapy,can achieve more productive and more targeted rehabilitation goals.
7.THE STRAIN SELECTION OF FLOCCULANT-PRODUCINGMICROORGANISMS USED TO SEPARATE NATURALSODA IN INNER MONGOLIA
Wen-Yu LU ; Tong ZHANG ; Dong-Yan ZHANG ; Chun HAN ;
Microbiology 1992;0(02):-
To solve separating problem of natural soda, fifty-seven strains screened from soil, floul water and activated mud were of flocculating activity. Two strains of bacteria, which were screened from above mentioned strains have higher activity and better steady than the whole culture liquid of bacteria was observed that its flocculating use to natural soda was strong and the mean flocculating rate of two strains were 79.80% and 87.% respectively.
8.Estimation of renal artery stenosis with MR IFIR-FIESTA renal artery angiography: a comparison with CT angiography
Xiaohui YIN ; Yan HAN ; Fengtan LI ; Renju BAI ; Dong LI
Chinese Journal of Geriatrics 2014;33(10):1081-1084
Objective To prospectively assess the diagnostic accuracy of MR inflow inversion recovery (IFIR) fast imaging employing steady-state acquisition (FIESTA) for detection of renal artery stenosis (RAS),as compared with renal artery CT angiography (CTA) performed as the reference standard.Methods Thirty patients [18 males,12 females; mean age (65.0±12.1) years] with arterial hypertension and suspected RAS were examined by 3.0-T 3D MR IFIR-FIESTA and renal artery CTA within a week.The IFIR-FIESTA image quality,the IFIR-FIESTA and CTA in display of second-order and third-order branches of renal artery were assessed by two experienced readers.The sensitivity,specificity,accuracy,and negative predictive value (NPV) of IFIR-FIESTA were calculated on artery-by-artery and patient-by-patient bases by the two readers.The inter-observer agreement between the two readers in determining the presence and grade of renal artery stenosis was assessed.Results The scores of qualification of IFIR-FIESTA image quality by reader A and B were (2.16±1.02) and (2.00 ± 0.14),there was no significant difference between them (Z=-0.037,P=0.971).IFIR-FIESTA and CTA in display of second-order and third-order branches of renal artery had no significant difference (x2 =0.298,P=0.585).In the assessment of all 64 main renal arteries by reader A,IFIR FIESTA on artery-by-artery basis had sensitivity,specificity,accuracy,and NPV of 100%,96%,97%,and 100%,respectively; the above parameters assessed by reader B were 100%,90%,93%,and 100%,respectively.There was excellent inter-observer agreement (Kappa=0.803) with the presence or absence of hemodynamically significant RAS.In the assessment of all 30 patients by reader A,IFIR-FIESTA on patient-by-patient basis had sensitivity,specificity,accuracy,and NPV of 100%,95%,97%,and 100%; the above parameters assessed by reader B were 100%,85%,90%,and 100%,respectively; there was good inter observer agreement (Kappa=0.724) with the presence or absence of hemodynamically significant RAS.Conclusions Compared with CTA,MR IFIR-FIESTA can excellently assess RAS without contrast material and radiation exposure,therefore,it can be used for screening of RAS,especially should be widely applied to elderly patients with arterial hypertension.
9.Correlation study of small vessel disease caused acute lacunar infarction and urine microalbumin
Zhiyong ZHAI ; Yan GAO ; Dong HAN ; Juan FENG
Chinese Journal of Cerebrovascular Diseases 2014;(9):480-484
Objective Toinvestigatethecorrelationofsmallvesseldisease(CSVD)causedacute lacunarinfarctionandurinemicroalbumin.Methods Theclinicaldataof136patientswithacutelacunar infarction admitted to the Department of Neurology,Shengjing Hospital of China Medical University from November 2012 to April 2014 were analyzed retrospectively. They were divided into either a CSVD group (n=72)or a cerebral large vessel disease (CLVD)group (n=64)according to their carotid artery color Doppler ultrasound and head magnetic resonance angiography findings. The levels of urinary microalbumin in both groups were observed and compared. SAS 9. 1 software was used to conduct statistic analysis. A Logistic regression analysis was used to determine the independent risk factors for CSVD caused acute lacunarinfarction.Results TheconcentrationofurinemicroalbuminoftheCSVDgroup(22±13mg/L) was significantly lower than (29 ± 14 mg/L)that of the CLVD group. There was significant difference (P<0.05). There was significant difference in the increased urine microalbumin levels between the CSVD group and the CLVD group (P<0. 01). There was an increasing trend for the proportion of patients with urine microalbumin concentration 10- <30 mg/L (56. 9%[41/72])in the CSVD group compared with the CLVD group (26. 6%[17/64]). Logistic regression analysis showed that the slightly increased microalbuminuria was associated with CSVD caused acute lacunar infarction (OR,3. 130,95%CI 1. 481-6.618;P<0.01).Conclusion Theslightlyincreasedmicroalbuminuriaisanindependentriskfactorfor CSVD caused acute lacunar infarction.
10.Association of single nucleotide polymorphism of xeroderma pigmentosum group C and smoking with genetic susceptibility to pancreatic cancer
Tieying HE ; Dong YAN ; Xiyan WANG ; Qilong CHEN ; Hai LIN ; Wei HAN ; Yan LI
Chinese Journal of Digestive Surgery 2013;12(8):581-585
Objective To investigate the correlation of single nucleotide polymorphism of rs3731055 and rs2607775 of xeroderma pigmentosum group C (XPC) and smoking with genetic susceptibility to pancreatic cancer.Methods The clinical data of 214 patients with pancreatic cancer who were admitted to the First and Third Affiliated Hospitals of Xinjiang Medical University from January 2009 to June 2011 and 214 healthly individuals were retrospectively analyzed.The samples of venous blood of 214 patients with pancreatic cancer (case group) and 214 healthy individuals (control group) were analyzed by the Multiplex SNaPshot method.The count data were analyzed using the chi-square test.The association between the single nucleotide polymorphism of rs3731055 and rs2607775 with genetic susceptibility to pancreatic cancer was analyzed using the Logistic regression method.Results Four hundred and twenty-three samples of gene were successfully typed,including 210 in the case group and 213 in the control group.The frequency of G allele of XPC rs3731055 was 75.95% (319/420) in the case group and 77.00% (328/426) in the control group,with no significant difference between the 2 groups (x2 =0.12,P > 0.05).The frequencies of genotypes GG,GA and AA were 58.57% (123/210),34.76% (73/210) and 6.67% (14/210) in the case group,and 60.09% (128/213),33.80% (72/213) and 6.10% (13/213) in the control group,with no significant difference between the 2 groups (x2=0.12,P > 0.05).The frequency of C allele of XPC rs2607775 was 87.86% (369/420) in the case group and 93.43% (398/426) in the control group,with significant difference between the 2 groups (x2=7.75,P < 0.05).The frequencies of genotypes CC,CG and GG were 77.62% (163/210),20.48% (43/210) and 1.90% (4/210) in the case group,and 86.85% (185/213),13.15% (28/213) and 0(0/213) in the control group,with significant difference between the 2 groups (x2=8.54,P < 0.05).Patients with rs2607775 GC genotype were associated with a significantly increased risk of pancreatic cancer compared with patients with rs2607775 CC genotype (adjusted OR =1.81,95% CI:1.06-3.10,P < 0.05).Patients with rs2607775 GC + GG genotype were associated with a significantly increased risk of pancreatic cancer compared with patients with rs2607775 CC (adjusted OR =1.98,95% CI:1.16-3.36,P < 0.05).The ratio of patients in the case group who smoked cigarettes ≥ 17 pack years was 25.24% (53/210),which was significantly higher than 13.15 % (28/213) of the control group (x2 =11.37,P < 0.05).The results of univariate analysis showed that patients who smoked cigarettes ≥ 17 pack years had higher risk of getting pancreatic cancer (adjusted OR =2.82,95% CI:1.27-6.29,P < 0.05).Patients who smoked cigarettes ≥ 17 pack years and with rs2607775 CC also had higher risk of getting pancreatic cancer (adjusted OR =2.87,95% CI:1.18-6.99,P <0.05).No significant gene-environment interaction was observed between rs2607775 GC + GG and smoking ≥ 17 pack years (adjusted OR =3.65,95% CI:0.67-20.03,P > 0.05).Conclusions The polymorphisms of XPC rs2607775 may play a role in the onset of pancreatic cancer.Patients who smoke cigarettes ≥ 17 pack years are more easily to have pancreatic cancer.There is no interaction between smoking and XPC rs2607775 in influencing the progression of pancreatic cancer.